I Denartrn^nt
A LABORATORY GUID
ill
Pharmac,
Univers
•
Torondo
PHARMACOLOGY
TORALD SOLLMANN, M. D.
Professor of Pharmacology and Materia Medica in the School of
Mrdicinr of \\Vstern Reserve University, Cleveland
ILLUSTRATED
I'llll.M.n.l'HIA AND LONDON
\\ B. SAUNDERS COMPANY
1917
logy
ty
Copyright, 1917, by W. B. Saunders Company
THE USE IN THIS VOLUME OP CERTAIN PORTIONS OP THE
TEXT OP THE UNITED STATES PHARMACOPOEIA IS BY VIRTUE
OP PERMISSION RECEIVED PROM THE BOARD OP TRUSTEES
OP THE UNIT-CO STATES PHARMACOPOEIA!. CONVENTION. THE
• AID BOARD OP TRUSTEES IS NOT RESPONSIBLE POR ANY
INACCURACY OP QUOTATION NOR POR ANY ERRORS IN THE
PRINTED IS AMERICA
PRESS OF
W. 8. SAUNOERS COMPANY
PHILADELPHIA
1
Departtr
Pharmac
University
of
PREFACE
Tin: following exercises are designed to introduce the student personally
to some of the more important facts of pharmacology. They have been
selected so as to present little difficulty to one versed in ordinary chemic and
physiologic technic, and require but little help on the part of the instructor.
The pharmaceutic and toxicologic exercises (Part I) are confined strictly
to the bare essentials needed by students who intend to become general
practitioners of medicine. Especial stress has been laid on the facts which
have a direct practical bearing.
The experiments on animals (Part II) have been arranged in groups, to
illustrate various types or phenomena, to bring out the similarities and dif-
ferences of the response of organs to pharmacologic agents, rather than by
individual drugs. This arrangement articulates better with the student's
experience in physiology and pathology, on which pharmacology is largely
founded. It is, therefore, more natural, as well as more interesting and in-
spiring. It has been the practice of the author to have this experimental
course precede the didactic course. The latter, dealing with individual
drugs, can then be based upon phenomena with which the student is already
familiar.
The exercises are arranged for a course of thirty working periods of two
to three hours. Additional experiments, for longer courses, demonstrations
.ire introduced as optional. They can, of course, be indefinitely extended
by t he use of dose tables and of the "Technical Notes." These are intended
primarily for the instructor and in\t-t iuMt<>r, indicating the sources where
more detailed information and different methods may be found.
CONTENTS
Department
Pharmacology
University
ol
Toronto
PAGE
INTRODUCTION 17
PART I
CHEMIC EXERCISES
CHAPTER I
GENERAL REACTIONS OF PLANT CONSTITUENTS 35
CHAPTER II
PHARMACEUTIC PREPARATIONS AND DISPENSING 41
CHAPTER III
MPATIHILITY 46
CHAPTER IV
ISOLATION OF POISONS 49
CHAPTER V
SPECIAL TESTS OF IMPORTANT ALKALOIDS 54
CHAPTER VI
\i. TESTS FOR IMPORTANT GLUCOSIDS AND \i ITKAL TRIM IPLES 60
CHAPTER VII
I\L TESTS OF IMPORTANT AROMATIC DERIVATIVES. 62
CHAPTER Mil
I KSTS FOR IMIMIKI-XNT Anrnvrn h 66
CHAI'TI K IX
OF IMPORTANT HEAVY METALS. . 73
CHAP I!
SPECIAL REACTIONS OF EARTHX \\u AIKUI Mi IMS
CHAPT1 K \i
CAUS \L ACIDS AND ALK\ 79
CIIAI'M K XII
\L REACTIONS OF INORGANIC Aero RADICALS. . 70
!4 CONTEN
CII Al'II.k XIII PAGE
FLAVORS . . 84
CHAPTER XIV
on
COLORS. ss
Lk XV
CHEMK STUDY OP u OF DRUGS is MAN ........................ 90
l HAMI.k XVI
CHEMIC ANTIDOTES ................................ 95
XVII
ADSORPTION BY COLLOIDS — ................................ 97
UIAPTER XVIII
SELECTIVE SOLVENTS ....................................................... 98
CHAPTER XIX
OSMOSIS AND DIFFUSION .................................................... 101
CHAPTER XX
DETERMINATION OF MOLECULAR CONCENTRATION ............................. 105
CHAPTER XXI
AGGREGATION OF COLLOIDS ................................................. 108
CHAPTER XXII
HEMOLYSIS, CRENATION, AND AGGLUTINATION OF RED BLOOD-CORPUSCLES .......... 109
CHAPTER XXIII
.................................................... in
CHAPTER XXIV
EFFECTS OF DRUGS ON HEMOGLOBIN ......................................... 113
CHAPTER XXV
CHEMIC EFFECTS OF CORROSIVES AND I KRITANTS .............................. 115
CHAPTER XXVI
PHYSIOLOGIC EFFECTS OF IRRITANTS ........................................ 119
CHAPTER XXVII
CXTH \RTH - OH MAN ....................................................... 121
CHAPTER XXVIII
ANTISEPTICS ............................................................. '. 121
CHAPTER XXIX
EFFECTS OF DRUGS ON FERMENTS. . . ........ 124
CONTENTS 15
I IIAI'II-R XXX
PAGE
•CELLULAR ORGANISMS AND Li 1:7
UIAI'TKk XXXI
A.N THELMINTICS AND INSECTICIDES I2Q
I ART II
EXPERIMENTS ON ANIMALS
CHAI'TI.k XXXII
LOCALIZATION OF ACTIONS; M INHI \\ i> \\i> DEPRESSANTS 132
CHAI'Tl.k XXXIII
; I.\K CONTRACTION : SKI.I.I i \i MusctE, C'n.ix 148
( IIAITI-k XXXIX
SMOOTH Mix 1.1.: IMI.MIM, QlKRUS, \M> AKTIRIKS 159
(II M'lKk XXXV
REACTIONS OF BLOOD-VESSELS (PERFUSION EXPERIMENTS, ETC.) 167
CHAI'TI.k XXXVI
EXCISED AND FROG HEARTS 182
CHAI'TI k \\XVII
AUTONOMIC DKI«.- \ E>UPTLS; (B) GIANDS; (C) BRONCHIOLES; (D) ANAFBYLAXJS;
(E) Kxrn \TIVI. INI i. \M\I\ rn»N :o:
CHAF1 1 k xxxvili
: i-:\s\ ; i-;\n ncs \ \a# '< i'i (ON; B) 1 . (C)
DI N AMD IMTEBACTION OF Dsooi; i» [moevNCR/ >PIN THY-
ROID B; (F) ANTEMETK
CHAI'II k XXXIX
MI i\it'>n-\i. i >; i \ rncpnATirRB; (B) GLYCOSUVIA; (C)
MM i» CENTRA] i" am TUATMENI "i I>M'KI»\NT
I'" ' KXTTS OF
IRKI
( II \ri i i: XI
i MM i NT OJ I'
CHAPT1 k XI i
HLOOD-PRESSURI
( II \l'l I l: XI II
ADMI
1 6 CONTENTS
Ul \MT.k Xl.lll pAGE
YASOMOTOR DRUGS; TREATMENT OF CIRCULATORY COLLAPSE 265
(II AFTER XLIV
OUNCES IN HEART-RAIL. 1C TV ... 279
CHAPTER XLV
M YOCARD1AL DEPRESSANTS AND TONICS 284
CHAPTER XLVI
INTESTINAL OSMOSIS— DIURESIS— TREATMENT OF ACUTE CARDIAC LESIONS 289
APPENDIX
APPENDIX A.— ARRANGEMENT AND GENERAL EQUIPMENT OF LABORATORIES 297
APPENDIX B.— EQUIPMENT OF CHEMIC LOCKERS (FOR EACH PAIR OF STUDENTS) . . 298
APPENDIX C. — REAGENTS NEEDED FOR CHEMIC EXERCISES 299
APPENDIX D. — CONTENTS OF LOCKERS FOR PHARMACODYNAMIC EXERCISES 304
APPEM : ! Ul'HABETIC LlST OF SOLUTIONS NEEDED FOR PHARMACODYNAMIC
KCISES 305
APPENDIX F. — TABULATION OF ANIMALS REQUIRED FOR DEMONSTRATIONS AND FIVE
GROUPS OF STUDENTS 309
APPENDIX G. — SOLUTIONS AND MATERIALS NEEDED FOR INDIVIDUAL PHARMACODY-
NAMIC EXERCISES 310
APPENDIX H.— DOSES FOR ANIMALS 320
339
To
A LABORATORY GUIDE IN PHARMACOLOGY
INTRODUCTION
The Objects and Methods of Laboratory Instruction. — It seems quite
superfluous at this time to insist on the great value of laboratory instruc-
tion. It may be well, however, to summarize the objects which it must
keep in view. These consist in imparting information, in developing an
understanding of the subject, and in acquiring a technical training. The
information which can be derived directly from laboratory work forms
the proper basis of didactic instruction: It facilitates the understanding of
those facts which are deduced from experiments; it illustrates their value
and their limitations; it impresses them on the memory. The training of
a laboratory course cultivates manual dexterity and, what is more im-
portant, it fosters the "scientific spirit" — the judicial attitude of mind
which requires the objective demonstration of statements and theories, and
which deduces from these objective data the conclusions which they justify
—no more and no less. The ultimate goal of this instruction should be
to enable the student to deal critically and independently with the matter
which is presented to him; to give him a more vital grasp of the whole sub-
ject of pharmocologic knowledge; and to generate and stimulate a healthy
thirst for further information.
The course of instruction which will meet these requirements in the
best attainable manner must vary somewhat with the resources at the
command of the department; with the size of the classes; and with the
special qualifications of the students and instructors. This applies par-
ticularly to the total time which can be devoted to laboratory work, and its
apportionment to class demonstrations and to individual work by the
students. The most thorough training would probably be obtained if the
student were to perform every experiment for himself, with a minimum of aid
from the in-tructor. The time which would be required for this purpose is,
however, quite prohibitive; nor is this plan essential. Demonstrations —
urnr u h a manner that every student can see the experiment, and
so that a- many as possible may assist in its performance — are almost as
ml- the information acquired, and can be substituted for a
considerable number of individual experiments in regard to the training:
especially if the -Indent ha- him-elf performed similar experiments. They
replace individual work completely, and as much of this
should be given as time and material permit. The demonstrations are
'.ntagemi>ly shown in connection with the individual laboratory \\
rd from their experiment - to \\ati li the rc-nlt- of the
demonstration^. This eomomi/e- time \\hen lengthy preparation or inter-
mittent observations are involved; it facilitate- the co-operation of the
student- and demonstrator-; and it emphasizes the close relation of
demon-it itioi^ and of the individual \\orL Another expedient of econoim ,
l8 A LABORATORY GUIDE IN PHARMACOLOGY
which is extensively utilized in this course, consists in having parts of the
class perform analogous experiments, but with different drugs; the results
of each section being demonstrated and reported to the entire class. A
great deal of time can also be saved by having the apparatus and reagents
in good order, systematically arranged, and conveniently accessible. The
student should "co-operate in this by keeping his working-place orderly,
neat, and clean.
:i with the closest management of the time it is naturally impossible
to present every possible pharmacologic experiment to the class. Those
experiments should be selected which demonstrate fundamental facts and
methods in the simplest manner. Experiments which consume much time,
or which are beset with special difficulties, or which are so exposed to acci-
dents that they are more apt to fail than to succeed in the hands of ele-
mentary students, are not suited to the conditions of an ordinary labora-
tory course, and may be left to advanced students who wish to devote extra
time to the subjects.
The mere performance of these experiments has only a very limited
value if the student does not study them exhaustively. He should have
a definite conception of the object of each experiment before he undertakes
its performance; and he should render to himself an account of every step
of the process, and of the conclusions to which it leads. The student's
note-book is therefore a very essential part of the course. Nothing culti-
vates the powers of observation like the taking of careful, detailed notes
during the progress of the experiment; while the critical faculty is stimu-
lated by the condensation of these detailed results into brief and definite
conclusions. This applies particularly to the animal experiments. The
constancy or variability of the results are illustrated by comparing the
results of different members of the class and of preceding classes. For this
purpose it is well to appoint a class reporter for each exercise, with the
duty of collecting and comparing all the results; these reports being kept on
file for the use of succeeding classes. They should be read and discussed
in the laboratory conferences.
Teachers differ in opinion as to whether the objects of the experiments
and the expected results should be pointed out to the student in advance.
In a pharmacology course the author believes that it is more useful to do so,
on account of the complexity of the subject, and the large ground which has
to be covered.
Relation of the Laboratory and Didactic Instruction. — The laboratory
course may be treated either as an adjunct to, or as the basis of, the didactic
instruction. If it is intended to illustrate the didactic teaching, it should
keep step with the latter; the experiments should be arranged with reference
to each drug. In the author's opinion, however, the course is much more
valuable if it is made the basis of the pharmacologic instruction ; if it is used
to deduce the facts rather than to illustrate them. For this purpose the
laboratory course should precede the didactic instruction ; and the exercises
should be arranged with a view to the pharmacology of particular organs, and
the methods used in their investigation, rather than with regard to the
individual drugs. If the conclusions are correctly drawn and summarized
the student will enter on the didactic study with a fairly extensive, first-
hand knowledge of the principal facts. The purpose of the didactic instruc-
tion will then be to correlate, apply, and extend these facts..
An elementary laboratory course is, of necessity, somewhat unevenly
balanced. It is much better suited for the development of some facts than
INTRODUCTION 19
of other?: and undue stress seems therefore to be placed on the former.
The "explanatory notes" and the "introductory discussions" are inserted
to meet this objection. These are made as elementary as possible to keep
them within the scope of the experimental knowledge of the student.
Even with these, however, it is impossible at times to avoid an exaggera-
tion of the laboratory side of the subject, and a comparative neglect of
features which may be of greater practical therapeutic importance. This
drawback should not be vital, for the didactic study should restore the
balance. Attention should also be called to this subject by the demonstra-
tors whenever necessary.
General Remarks on Note Taking. — The results of the experiments
should be entered briefly in a special note-book. The method should be
indicated sufficiently to make the notes understandable. Tracings should
also be inserted when possible; either the original, or copies taken free hand,
with tracing-paper, or blue prints. Unnecessary detail is to be avoided.
The results should be followed by a brief statement of the conclusions which
may be drawn from the experiment. These should only bear on principles,
not on details. They should go no farther than the data of the experiment
warrant. The "Questions" at the end of the Exercises may guide in this.
.Students should always read the experiments before coming to the class.
This is especially important when animals are to be used.
REFERENCE BOOKS
The following will be found useful in the laboratory, particular!
the details of methods:
Abderhaklen. — Handbuch der Biochemischen Arbeitsmethoden, Berlin.
Association of Official Agricultural Chemists. — Methods of Analysis,
United States Dept. Agric., Bur. Chem., No. 107.
Autenrieth. — Detection of Poisons, translated by W. H. Warren.
Edmunds and Cushny. — Laboratory Guide in Experimental Pharma-
cology, Ann Arbor, 1905.
Fuehner. — Nachweiss und Bestimmung von Giften auf biologischen
Wege, Berlin, 1911.
Gadamer. — Lehrbuch der chemischen Toxicologie, Goettingen, 1909.
Gooch. — Methods in Chemical Analysis, New York, 1912.
cene. — Experimental Pharmacology, Philadelphia, 1909.
Harvard Apparatus Co. — Catalog.
Hatcher and Sollmann. — Text-lxx>k of Medica, Philadelphia,
1004.
Hein/. Handbuch der operimenteUen Pathologic und P' -logic,
a, 1905.
Hoeber.— Physikalische Chemie der Zellen und Gewebe, Leipzig.
Robert.— Lehrbuch der I nt ..\icationen, Stuttgart. 1902.
(Quantitative BeMimmunu' der Alkaloid,-. Berlin. 1013.
Lenhartz. — Mikroskopie und Chen krankenbett. Berlin, 1910.
KII Yer/eii hniss, Berlin, 1913.
ional Formulary.
Nelson.— Analysis of Drugs and Medicines, New York, 1910.
Pharmacopoeia of the United States.
Bio, iladelphia, 1914.
Sahli. Diagnostic Methods. Philadelphia.
•man. Organic A
Sollmann. Manual of Pharmacology, Philadelphia.
20 A LABORATORY CU1DK IN PHARMACOLOGY
Stewart.— Manual of Physiology. New York.
Sutton.— Volumetric Analysis. Philadelphia.
^erstedt.— Handbuch der Physiologischen Methodik, Leipzig.
Wester.— Darstellung phytochemischer Uebungs-praeparate, Berlin,
19 1
SCHEDULE OF COURSES
The following detailed outline of the pharmacologic courses given in the
author's laboratory may offer helpful suggestions:
COURSE L -ELEMENTARY PHARMACY, GENERAL TOXICOLOGY, AND PRINCIPLES
OF PRESCRIPTION WRITING
Two hours laboratory and one hour of didactic instruction per week
in the first semester of the second year. Students work in pairs.
The numbers in the following refer to the weeks; (a) to the one-hour;
(b) to the two-hour periods. "Optional" experiments are omitted.
(la) Lecture and Demonstration: Pharmacognosy and Plant Constit-
uents.
(ib) Laboratory: Assignment of Lockers. Reactions of Plant Con-
stituents, Chapter I.
(aa) Lecture and Demonstration: Pharmaceutic Methods; Assaying.
Recitation: On Lecture la.
(20) Laboratory: Pharmaceutic Preparations, Chapter II, Exercise I
to VIII.
(3a) Lecture: Liquid Pharmaceutic Preparations.
Recitation: On Lecture 2a.
(30) Laboratory: Pharmaceutic Preparations, Chapter II, Exercise
VIII to XIV.
(4a) Lecture: Solid Pharmaceutic Preparations; Solubilities.
Recitation: On Lecture 3a.
(40) Lecture: Incompatibilities.
Laboratory: Incompatibilities, Chapter III, Exercise I to III.
Lecture and Demonstration: Metrology.
Recitation: On Lecture 4a.
(50) Recitation: On Lecture $a.
Laboratory: Incompatibilities, Chapter III, Exercise IV to VI.
(6a) Recitation: Incompatibilities.
Review: Metrology.
(6b) Lecture and Demonstration: Toxocologic Analysis and Assaying,
Chapter IV.
(ya) Recitation: On Laboratory 6b.
Review: Incompatibilities and Solubilities.
(yb) Laboratory: Tests for Important Drugs, Chapter V to VIII.
(8a) Written Test on Text and Laboratory "Questions."
Assignment of experiments and reporters for Exercise XIII.
(8b) Laboratory: Flavors, Chapter XIII (part).
Excretion of Drugs, Chapter XV (part).
(pa) Lecture: Treatment of Poisoning.
Prescription Writing.
(90) Conference: On Laboratory 8b.
Laboratory: Flavors, Chapter XIII (part).
Excretion of Drugs, Chapter XV (part).
INTRODUCTION 21
(ioa) Lecture: Flavors and Colors.
Recitation: On Lecture oa.
(icb) Conference: On Laboratory gb.
Laboratory: Colors, Chapter XIV. *
Excretion of Drugs, Chapter XV (part).
Prescription Practice (in sections, alternating with
laboratory work).
Study Materia Medica Lessons i and 2.
(na) Recitation: On Lecture ioa and Materia Medica Lessons i and 2.
Conference: On Laboratory lob.
(nb) Laboratory: Excretion of Drugs, Chapter XV (finish).
Chemic Antidotes: Chapter XVI.
Prescription Practice.
Study Materia Medica Lessons 3 and 4.
(i2a) Lecture: Treatment of Disease; Chemical and Physical Basis of
Pharmacology.
Recitation: Materia Medica Lessons 3 and 4.
Conference: Laboratory nb.
(i2b) Laboratory: Absorption and Selective Solvents, Chapters XVII
and XVIII.
Prescription writing Practice.
(i3a) Lecture: Manifestations of Pharmacologic Action.
Recitation: On Lecture i2a.
(i3b) Conference: On Laboratory i2b.
Laboratory: Osmosis, etc., Chapter XIX to XXI.
Prescription Practice.
(i4a) Lecture: Administration of Drugs.
Recitation: On Lecture i3a.
(i4b) Conference: On Laboratory i3b.
Laboratory: Hemolysis, Irritants, etc., Chapter XXII to XXVI 1
Prescription Practice.
(i$a) Lecture: Conditions Influencing Drug Actions.
Recitation: On Lecture i4a.
(i5b) Conference: On Laboratory i4b.
Laboratory: Antiseptics, Ferments, etc., Chapter XX VI II to
XXXI
Prescription Practice.
(i6a) Recitation: On Lecture i5a.
Conference: On Laboratory i5b.
(i6b) Written Te>t on I :>tion Writing and Laboratory
"Questions," Identification of Spo inn
Assignment of Lockers for Animal Work.
COURSE II. -EXPERIMENTAL PHARMACODYNAMICS
Time hour- «.f lal>oratory work and two hours <>: nces per
in tl the -eeond year. Student^ \\otk in groups of
<li\i<le<l int.. subgroups A and B. The cxj>- arc arranged for
five full groups.
The -yllal.u in the following table, which also gives the group
and number (A to F) of the student- \\li.. act as class reporters for each
periment. It i- their duty t lle« t tlie individual reports and present the
significant rc-ult- at the "* "iifcTences."
22
A L.\BOR.\TORY GUIDE IN PHARMACOLOGY
•SL *° JS
I > T
E « •£
£ bo
1 1
<£
M QV
•/.
£
01
»0
1
W
o
3
0
1
to
•
CO
IV
Exercise I,
C
1
4
Exercise V,
in- 5. Cocain,
sium.
1
-
to
hH ^
*
CO
*3
SH-~
c3
>"
rf
'eS
|
i
a
p
ercise
Ii
2
u
£
CO
x
H
10
4
d
1
tO
CO
o
to
Ht-T
"o
>
g"
•^
1
J3
1
"§
u
j-Sj
Q
pS
1
cl
1
. u
to
CO
s
|
gc^
vo
<U HH ^
|
B
o
x
W
•C^'C $'C c o
IIP 11 a
W W W H
v ^ 5
«! ijwl i<^
frt ji^sSK
I S & — x»-rS bH h t-
?;|w U^^X^K^^"
J^
5
. •? f
I > I _
c/o I" pj
t^ W 4
4. Magnesium.
5. Physostigmin
Exercise V, 3.
4. Cocain-quini
4. Nicotin.
5. Pstig
Exer V 3.
ysost
ise V
ain-s
.§-5
Mi
«i 1/3 rj Kf
IIP;
w w w
.s I Is Is a
u d w C <" C <u
W W W
INTRODUCTION
6|
^
2
I S
W 4
<
«0
•s «
n
£
y— «.
s2
M
1
! !
d
1
d
•0
1^
s
X
X
!
(z) Local Irritati
i. (10) Barium.
Ill
Pilocarpin-atro]
j
5
a
\
' =.
I I
§> &
A 1. \BORA TORY GUIDE IN PHARMACOLOGY
INTRODUCTION
••:•'-
I ^
! I
;i
..cr
|«j <<|-gr-j £ i" rs2.!0.'!-.-!^-:
if .sj%! .2 Jutripj
vr«uU'vS'vrvrv rn -«• l'fl'rtaj?r£B^:'rta:3^ 1 « .Vtt _C • Jr tt if a J9
26
A L.\BOR.\TORY GUIDE IN PHARMACOLOGY
«1.p 1
-s Sll
1!
>
w X!
H
W
>55
| ^
C/3 *-"
I. „-§
s ^
H*
&. ^^
I tl^ <
^,sl> >
r- Ji fa CU w, fa
-pis!
X <u S m 5 <u
c(*£^2tf
g^g^-c^-
H U £
INTRODUCTION
?i
u
^ -
— C/2
i i
I
I*
=11 .
slo-dt
^5 ~£ = 2,-i
KJ
;
28
A LABORATORY GUIDE IN PHARMACOLOGY
SiSiag II
•§•=
r
o w
d I .
} II
Si
S ^|
•* g rf
S 'i| 1
B 1 of
•o .3 S &S. .
«g£>. ^g=2^§
It S|c3e|
i us! § ip gy
.I/JW. . .»H. O'™^1-1'*"^^^
^•10 \Ot^OOO*MM
WU
SI
g
1
•£ &
•5 «|^ :fc g
2 O^J < §
< S§« * 2
g -
2 £3$!!
c.Si
)2W M o 5>SgǤl a
!§• l|] Ijljldj
INTRODUCTION
29
><8f
lit
i
-fij |5
l
HI t! I 11 If
^ < X — y. U _ '_ < 7
M cJ f*j «f lA. >c t^. . O» O « e* to
-
«T
3
_=
"H
1
£
-oQ ., 2
^185,
£ = --
Si £•= "5
C> 0
'i.g fr
sg = s
O tfj ^>
Sail
JE-H.^-= sil*2
"?.5 5-^ = 3-^E =
-~'E.^t rE£^
7/-'-^ - ^ < /
x ci rr> ^ to O t^OO
£ •
H i<
12 5 *F->.C-=O
^•^ £'E-5..5c5 x-2
"9 O jj e/: < ^ — ^ — —
rnin.
My!
^J i
^ ^
1
do -
A LABORATORY GUIDE IN PHARMACOLOGY
^ ^ !i
^ n
i8B3«2sJ« J5i
I
i i
j b
O -/5 .
- -ij fe
-J z!j 11§ I"!
^^ C5 = ^-~ <«
i O M-44 4^>
v°
I c
Ci
JI
- ^ • J
li- 1
-.^ I .
'— • / — —
a-o ^co
!.l|>
c •** - ^ 'x -
"lliiJ
^ M. B^«U
O « «' r^
Urine.
"" •- = -1 ^rl c
I!
II
cart-ra
sure a
sions
Weak
Maxi
Cev
IJli
c^U>co
*jU
l§
2
^
S-J «
1—1 '-2 c
Jg
C/3 ^
j5i
&IJ
.gjg
rt *fcr ^H
it
>» cj X!
EUU
Ha
u
M C^
ro •* to
lg
|l
s ^
« s
§•1
"5'S
CCA)
INTRODUCTION
32 I_\BORATORY GUIDE IN PHARMACOLOGY
COURSE III. -SYSTEMATIC PHARMACOLOGY: DRUGS WITH PREDOMINANT LOCAL
ACTION
One hour, lecture or recitation, per week in the fourth semisemester of
the second year.
1. Ferments and Nutrients.
2. Emollients and Demulcents.
( iencral Phenomena of Irritation, Corrosion and Astringents.
4. Inorganic Irritants and Astringents.
Irritant Volatile Oil>.
6. Irritant Volatile Oils and Physical Irritants.
• 7. Stomachics, Cathartics.
8. Cathartics, Anthelmintics.
COURSE IV. -SYSTEMATIC PHARMACOLOGY
DRUGS WITH PREDOMINANT SYSTEMIC ACTION
Four hours per week, didactic, in the first semisemester; and three hours
per week in the second semisemester of the third year. (L = Lectures;
R = Recitations.)
L i. Strychnin.
L 2. Caffein.
R i. On L i and 2.
L 3. Morphin.
L 4. Morphin, hydrastis.
L 5. Cannabis, cocain.
R 2. On L 3, 4, and 5.
L 6. Autonomic drugs.
L 7. Autonomic drugs.
L 8. Atropin, scopolamin.
R 3. On L 6 and 7.
L 9. Pilocarpin to curare.
L 10. Epinephrin, pituitary, thyroid.
L 1 1 . Ergot to nitrites.
R 4. On L 8 and 9.
L 12. Digitalis.
L 13. Digitalis.
L 14. Camphor to colchicum.
R 5. On L 10 and n.
L 15. Apomorphin to heat regulation.
L 16. Antipyretics.
L 17. Benzol antiseptics.
R 6. On L 12 and 13.
L 1 8. Benzol antiseptics.
L 19. Miscellaneous antiseptics, sera, vaccines.
L 20. Narcosis theories, alcohol.
R 7. On L 14, 15, and 16.
L 21. Alcohol.
INTRODUCTION
L 22. Anesthetics.
L 23. Anesthetics.
R 8. On L 17, 18, and 19.
Written tests and specimens, Lectures i to 19 inclusive.
L 24. Hypnotics, gases, cyanids.
R 9. On L 20 and 21.
L 25. Physics of salt action.
L 26. Physics of salt action.
R 10. On L 22, 23, and 24.
L 27. Physiology of salt action.
L 28. Cathartic salts, water, diuresis.
R ii. On L 25 and 26.
L 29. Cathions.
L 30. Anions.
R 12. On L 27 and 28.
L 31. Reaction.
L 32. Metals, inorganic compounds.
R 13. OnL 29, 30, and 31.
L 33. Organic arsenic compounds.
L 34. Antimony, bismuth, iron.
R 14. On L 32 and 33.
L 35. Radium, silver.
L 36. Mercury.
R 15. On L 34 and 35.
L 37. Lead, phosphorus.
R 16. On L 36 and 37.
Written tests and specimens on Lectures 20 to 37.
Examination on entire subject.
33
Universit
PART I
CHEMIC EXERCISES
Introductory Remarks. — Before beginning on the laboratory work
the student should check the contents of his locker and familiarize him-
self with the reagents on the shelves (see Appendix). These are arranged
alphabetically. Remember that they are to be replaced in their proper
position as soon as used. The student should supply himself with towel,
soap, matches, scratch-pad, and dissecting instruments. He should keep
his working-place clean and neat.
The experiments, explanatory remarks, and references should be assigned
and read before coming to the class. Cross-references to other experi-
ments (e. g., "Consult Exercise so and so") mean that these experiments
are to be read, but not to be performed, at this time. The student should
reflect on the object and conclusions of the experiment while it is in progress.
He should take account of all the experiments performed in the course,
including those shown as demonstrations or assigned to other members of
the class. Two students may collaborate in the chemic experiments.
Successful results should be checked in the book and the questions an-
swered in the note-book.
If an experiment is unsuccessful, it should be repeated. In the event of
a second failure, the student should call on the demonstrator for help.
unusual or atypical result should be reported.
Additional apparatus is furnished on written requisition. The special
material needed for each experiment is noted at the bottom of each page
(S. I/.).
CHAPTER I
GENERAL REACTIONS OF PLANT CONSTITUENTS
(It is assumed that the student is familiar with the characters of glucose,
cane-sugar, starch, proteins, and fats. Should this not be the case, they
should be studied before the following experiments are made.) Two
students may work together.
EXERCISE I.— ALKALOIDS »
1. Alkalinity.— Place a drop of i per cent, nicotin on red litmus paper:
blue color.
2. Precipitation Reactions. — Place on slides a few drops of i : 1000
acidulated quinin sulphate solution, mix with a drop of the following, and
note the amorphous precipitates:
(a) lodin in KI = Reddish.
(6) Mercuric Potassium lodid = White.
(Mayer's Reagent.)
(c) Picric Acid - Yellow.
(d) Tannin (about i per cent.) = Gray.
(e) Phosphotungstic Acid2 = White.
5. M— Nicotin, i per cent.
l.tr reactions are given by other organic bates, t. jr., pyridin aad quinolin.
* Phospholuntstic Acid: A 10 per cent, solution in 4 per cent. IK I
35
36 A LABORATORY GUIDE IN PHARMACOLOGY
3. Solubility Characters of Alkaloids and their Salts. — In a test-tube
make about 5 c.c. of an acidulated i : 1000 solution of quinin sulphate dis-
tinctly alkaline by XaOH solution: a precipitate of free alkaloid is thrown
down (free alkaloids are generally insoluble in water, while their salts are
soluble). Add about 10 c.c. of ether and shake with a gentle rotatory mo-
tion. Draw off the ethereal solution from the top with a pipet, and
again shake the watery solution with 5 c.c. of ether. Again draw off the
ether. Acidulate some of the remaining Avatery solution and test it with
mercuric potassic iodid, observing that there is no or very little precipitate
(the free alkaloid being completely extracted by the ether). Shake the
ethereal solution with some dilute sulphuric acid. Draw off a little of the
acid solution from the bottom, and test with mercuric potassic iodid: a pre-
cipitate occurs.1 (The acid converted the quinin into the sulphate, which
is soluble in water and insoluble in ether.)
4. (Optional) Lassaigne's Test for Nitrogen. — Place a knife-pointful of dry quinin
sulphate in a dry test-tube. Take a piece of metallic Na, size of small pea, dry with
blotting-paper, and add to quinin. Heat red hot and plunge into beaker with a little
water. Filter. Add a few drops FeSO4. Let stand five minutes. Acidulate with cone.
HC1 and heat: Greenish or blue color or precipitate of prussian blue.
Note the peculiar odor of quinolin, a decomposition product of quinin.
Explanatory Notes. — On heating with sodium, the N of quinin (and other nitrogenous
substances) gives sodium cyanid; treated with a ferrous salt, this gives the ferrocyanid;
on adding acid, this forms ferricyanid with the ferric salt formed from the ferrous sul-
phate.
5. (Optional) Microchemic Reactions.2 — Alkaloidal precipitates often present a
crystalline character, which may be useful hi their identification. This is illustrated by
the following examples. (Mix the solutions on a slide, and examine from time to time
with low-power microscope, until typical crystals are seen.)
(a) 5 drops of 2 per cent, morphin sulphate and i drop 10 per cent. NH4OH: rapid
formation of needles. (Rub with a glass rod if necessary.)
(6) 5 drops of ^ per cent, nicotin and excess of picric acid: at first a fine precipitate;
later stellate crystals.
(c) Substitute i per cent, atropin sulphate for nicotin in (6) : feathery crystals and
stellate groups.
(d) i per cent, strychnin sulphate and potassic bichromate solutions: fine rosettes of
needles at once.
6. (Optional) Preparation of Alkaloids. — Directions are given in D. H. Wester,
"Anleitung zur Darstellung phytochemischer Uebungspraeparate," Berlin, 1913. The
preparation of caffein and piperin are convenient examples. The preparation of individual
alkaloids is also described in Abderhalden's Handb., 2, 904.
QUESTIONS
(a) State the principal properties of alkaloids (reaction, precipitants,
solubility, characteristic element).
(b) How would you test a solution for the presence of alkaloids?
(c) Why is it necessary to apply several tests?
(d) How would you extract an alkaloid from a solution of its salts?
(e) Should alkaloids be prescribed with iodin or tannin?
TECHNICAL REFERENCES
Qualitative and quantitative tests, Abderhalden's Handb., 6, 118; Gadamer, Lehrb. d.
chem. Toxicologie.
i Water saturated with ether and acid may give a precipitate with Mayer's reagent, even in
the absence of alkaloids; but this non-alkaloidal precipitate dissolves on adding an equal volume
of water (A. H. Clark; reference, Amer. Jour. Pharmacy, 1909, 176).
»T. G. Wormley, "Microchemistry of Poisons," Philadelphia, 1885.
CHAP. I GENERAL REACTIONS OF PLANT CONSTITUENTS 37
EXERCISE n.— GLUcosros
1. Test a little fresh i per cent, solution of salicin (a glucosid) for re-
ducing sugar by Trommer's test1: negative.
2. Decomposition by Acids.— To another portion of the solution add
A volume of 10 per cent, sulphuric acid; boil in water-bath for ten minutes;
make alkaline with XaOH and apply Trommer's: positive.
3. Decomposition by Ferments.— To another portion of the solution
add some saliva and heat in water bath at 40° C. for half-hour; test for sugar:
positive.
4. Note difference in sweetness of alkaline and acidulated fluidextract
of licorice. (The sweet glucosid, glycyrrhizin, like many glucosids, is a
feeble acid, held in solution by ammonia and precipitated by strong acids.)
5. (Optional) Brunner-Pettenkofer's Reaction ((iiven by Glucosids and Sugars).—
Dissolve some glucosid and purified ox-bile in water, and pour carefully on a layer of
concentrated sulphuric acid: red ring at contact; on agitation the whole fluid is colored
red.
6. (Optional) Decomposition by Emulsin.— Preparation and tests of emulsin, Abder-
halden's Handb., 3, 391; 7, 760.
QUESTIONS
(a) What is the characteristic property of glucosids?
(b) How do they differ from ordinary carbohydrates?
(c) Why is it inadvisable to prescribe solutions of glucosids with acids?
(d) How would glucosids be affected in the alimentary canal?
(e) How would you distinguish between a glucosid and an alkaloid?
(f) How would you separate an alkaloid and a glucosid from a solution
containing both?
TECHNICAL REFERENCES
Preparation, Reactions, and Synthesis of Glucosids, Abderhalden's Handb., 7, 732.
EXERCISE in.— SAPONINS
Saponins give the typical reactions of glucosids. They lake blood cor-
puscles (see Chapter XXII).
i. Foaming.— Shake a few drops of a tincture of soap-bark i which is
rich in saponin) with a little water: considerable foam is produced, which
subsides slowly.
mulsification. — Add 25 drops of the soap-bark tincture to about an
inch of cotton-seed oil. Shake. Add an inch of water and shakr: a >mooth
mixture (emulsion) is formed. Add alcohol: the emulsion persists.
3. (Optional) Color Reaction.— C'«m«rntr;itnl -nlplimi. .1. id dissolves saponins with
a yellow to brick-red color, passing gradually through red to violet.
QUESTIONS
(a) How would you detect saponin in a plant extract?
(b) What is the « -\planation of the saponin action?
(c) What practical uses can be made of these actions?
per cent.; fluidcxtract limrirr. plain and acidulated.
1 Trommrrs Test ' >H and add dil< Iphatc. drop
by drop, until a «.liKht prrmam-nt pro i; ;TIC hydrate appear*. Boil:
yellow or l>r«- precipitate of cuprous oxid.
38 A LABORATORY GUIDE IN PHARMACOLOGY
TECHNICAL REFERENCES
Preparation, etc., Abderhalden's Handb., 2, 970; Detection in frothing liquids, etc.,
Gadamer, 446; Loncheux, ref., Yearbook Amer. Pharm. Assoc., i, 448, 1912; Determination,
Aoff, 1912, ref., fhem. Ahstr., 7, 803; Bio-estimation in drugs, Robert, 1912, ref.,
Yearbook Amer. Phann. Assoc., i, 446.
EXERCISE IV.-CATHARTIC EMODIN PRINCIPLES
1. Borntraeger's Reaction for Emodin or Chrysophanic Acid.— To an
infusion of rhubarb add a few drops of ammonia: red color.
2. (Optional) Hirschsohn's Reaction for Aloins. — Mix 10 c.c. of i : 1000 aloin solution
with i drop of 10 per cent, copper sulphate and of 2 per cent, hydrogen peroxid; boil:
red color (hindered by alcohol, acids, and alkalies).
3. (Optional) Stacy's Reaction for Aloes. — A delicate reaction with ferricyanid, the
tints distinguishing the varieties (Ref., Amer. Jour. Pharm., 88, 262, 1916).
QUESTIONS
(a) How would you determine whether a patient is taking an emodin
cathartic? (The chrysophanic acid passes into the urine ; however, santonin
and phenolphthalein urines give similar reactions.)
(b) What change would occur in a rhubarb urine on standing?
TECHNICAL REFERENCES
Assay of Emodin Drugs, E'we and Vanderkleed, 1913, Jour. Amer. Pharm. Assoc.,
2, 9795 Gadtamer, 422; Daels, 1913, ref., Jahrb. Pharmacie, 73, 6; Rhubarb, colorimetric
determination of value, Tsirch, 1904, Jahrb. Pharm., in; Detection of Emodin drugs in
presence of Phenolphthalein, L. E. Warren, 1914, Amer. Jour. Pharm., 86, 444; Determina-
tion of Drastic Purgatives, Gadamer, 426; Colocynthin, Test for, Venturoli and Vervi,
1009, ref., Jahrb. Pharm., 69, 587.
EXERCISE V.— TANNINS
(Dissolve a little tannin in hot water or use the i per cent, solution.)
1. Add drop of Fe2Cl6: green-blue-black color. Dilute until it is trans-
parent. Add a few drops of NaOH: garnet color. Add cautiously an ex-
cess of H2SO4: greenish-red; with more, greenish-yellow.
2. Add some Pb(C2H3O2)2: large white precipitate. Add NaOH and
shake: pink.
3. (Optional) Add some NaOH: reddish-brown color.
4. (Optional) Observe that tannin precipitates alkaloids (e. g., quinin), proteins
(egg-white solution), and gelatin.
5. Add a drop of Fe^le to a little infusion of Cinchona (greenish color).
The tannins occurring naturally in plants give a greenish color with iron;
tannins occurring in pathologic formations (nutgalls) give a bluish color.
6. (Optional) Gallic Acid. — To a i per cent, solution of gallic acid add a few drops of
i per cent. KCN: a red color appears, which soon fades, but reappears on shaking
(Young's test). Pure tannic acid does not give this reaction.
QUESTIONS
(a) How would you test for tannins in a plant-extract?
(b) What groups of substances should not be prescribed in solutions
with tannins?
(c) Why does tannin stop local bleeding?
(d) Why are tannin preparations useful in diarrhea?
5. M. — Rhubarb infusion, 5 per cent.
S. M. — Cinchona infusion, 5 per cent:
CHAP. I GENERAL REACTIONS OF PLANT CONSTITUENTS 39
EXPLANATORY NOTES
Ferric Chlorid as Group Reagent. — Ferric chlorid gives color reactions with a number
of organic drugs; for instance:
Red, with antipyrin; aliphatic amido-acids; meconic acid.
, with apomorphin; nirvanin; salicyl compounds; resorcin; phloroglucin; phlo-
rhizin.
Blue, with morphin; phenol; cresols; naphthol; hydroquinon; gallic acid; phenol-
sulphonic acids.
Green, with thallin; oxyquinolin; laudanin; epinephrin; pyrocatechin.
TECHNICAL REFERENCES
Isolation and testing of tannins, Abderhalden's Handb., 2, 096; 6, 146; Determination
in plant juices, ibid., 8, 259.
EXERCISE VI.— GUMS
(Use a 10 per cent, solution of acacia.)
1. Hydrolysis by Acids.— Test for sugar: negative. Add £ volume of
5 per cent, sulphuric acid, and boil for ten minutes in water-bath. Make
alkaline with NaOH, and test for sugar: positive. This test is given in
common by gums, starch, glucosids, and other carbohydrates.
2. Add some alcohol: precipitate (difference from glucosids; borax and
ferric chlorid also cause precipitation or gelatin ization).
3. Add a few drops of iodin solution: no blue color (difference from
starch) .
4. Note the viscosity of the solution; on shaking, it forms a rather per-
sistent foam. It emulsifies oils, although less readily than saponin.
QUESTIONS
(a) What are the characteristic properties of gums?
(b) Why are gums incompatible with tinctures?
(c) What would be the best menstruum for the extraction of gums?
(f) What menstruum would be used to obtain extracts free from gums?
(e) Explain the effect of gums on foaming and emulsification.
TECHNICAL REFERENCES ON CARBOHYDRATES
Abderhalden's Handb., 2, 43, 85, 119; <, 1385, 1408; 6, i; Starch, ibid., 6, i; Soluble
.S/.m/r, ibid., 6, 20; Samec and Jencic, Koll. Beih., 7, 137, 1915; Sugars, Abderhalden's
Handb., 2, 43, 85; 5, 1385, 1408; quantitative methods, ibid., 2, 167; in blood, ibid., 5
and Benedict, 1915, Jour. Biol. ('hem., 20, 61; Kahn, 1915, Jour. Amer. >
Assoc., 64, 241; Cellulose, AbderhaldmV Handb., 6, 28; in feces, 5, 378; Inulin ibid..
hit ion, ibid., 3, 218; Levulose, estimation in presence of glucose, Loewc. 1910,
Soc. Exp. Biol. Med., 13, 71.
EXERCISE VH.— RESINS
(Use commercial rosin.)
i. Solubility.— Note that this is soluble in alcohol, but is precipitated
from this solution by adding water. It i- also soluble in ether, turpentine,
ti\r«l oik an. I ln.iliiu; xxlium hydrate solution (precipitated by acids), but
insoluble in gasolin.
5. A/.— 10 per cent acacia.
40 A LABORATORY GUIDE IN PHARMACOLOGY
QUESTIONS
(a) State the important solubility characters of resins.
(6) What would be a good menstruum for the extraction of resins from
drugs?
Should resinous tinctures be mixed with waters?
(</) Explain the actions of alkalies and acids on resins.
(e) What would be the nature of the precipitate produced by nitric acid
in the urine of a patient taking resin of copaiba?
(/) How could you distinguish this precipitate from albumin?
EXERCISE VIIL— VOLATILE OILS
(Use oil of turpentine.)
1. Solubility.— Note that this mixes with alcohol, ether, gasolin, and
cotton-seed oil, but not with water. Camphor, which may be considered
as a solid volatile oil, behaves similarly.
2. Note that it makes a greasy stain on paper, but that this stain disap-
pears in time, especially on heating.
QUESTIONS
(a) State the solubility characters of volatile oils.
(b) Are volatile oils absolutely insoluble in water?
(c) What would be good menstrua for their extraction?
(d) What occurs if "spirits" are mixed with waters?
(e) How would you distinguish a volatile from a fixed oil?
TECHNICAL REFERENCES
Polarimctrk estimation of Camphor in Spirit, etc., Jahrb. Pharmarie, 69, 354; Deter-
mination of Camphor in urine, Abderhalden's Handb., 3, 975; Preparation of Volatile Oils,
ibid., 2, 982.
EXERCISE IX (OPTIONAL).— CHLOROPHYLL
i . Note the green color of a fresh tincture of lettuce leaves.1
\dd some dilute HC1: yellow color.
3. To another portion add some NaOH : the color becomes an old gold-green. (Chloro-
phyll has a characteristic spectrum, in which the above reagents produce definite changes;
see Hatcher and Sollmann; Materia Medica.)
TECHNICAL REFERENCES
Abderhalden's Handb., 2, 671; Preparation, Stanck, Chem. Abstr., 7, 1784; Lipochrome,
Abderhalden, 2, 723, 758; Animal Pigments, ibid., 2, 717.
FURTHER TECHNICAL REFERENCES ON PLANT CONSTITUENTS
Vegetable Proteins.— Abderhalden's Handb., 2, 270; Animal, ibid., 335; Removal,
ibid., i, 686.
Extract in Vegetable Preparations.— U. S. P. IX; Abderhalden's Handb., 8, 171.
Moisture.— U. S. P. IX; Abderhalden, 8, 167.
Methods of Rapid Desiccation of Tissues, etc.— Abderhalden's Handb., 5, 614;
\\ierhowski, 1007, Beitr. chem. Physiol., 9, 232; Shackell, 1905, Arner. Jour. Physiol., 24,
325; Beebe and Burton, 1005, ibid., 14, 9; Rosenbloom, 1913, Jour. Biol. Chem., 14, 27;
Lumiere and Chevrotier, 1912, Chem. Abstr., 7, 1521.
Mdting Point.— U. S. P. IX.
Boiling Point.— U. S. P. IX.
Congealing— U. S. P. IX.
Solubility.— U. S. P. IX.
Ash. — U. S. P. IX; Analysis, Abderhalden's Handb., i, 372; 5, 200, 1049; 6, 376.
Colorimeters. — Abderhalden's Handb., i, 642; Roberts, 1910, Hyg. Bui. No. 66.
Color Standards. — Amy and Ring, 1915, Jour. Amer. Pharm. Assoc., 4. 2294.
> Some fresh lettuce is bruised in a mortar with sand, triturated with alcohol, and filtered.
CHAP. II PHARMACEUTIC PREPARATIONS AND DISPENSING 4!
CHAPTER II
PHARMACEUTIC PREPARATIONS AND DISPENSING
Two students can collaborate on the experiments except those marked
"individual." To save time the solids may be weighed in advance by the
instructor. Some of the preparations will extend over several laboratory
days. The student should always start the day's work with these unfinished
preparations. The finished preparations should be submitted to the in-
structor, and can then be preserved in stock-bottles (for use in the later
exercises). The formulas of the optional preparations can be found in the
U. S. P. or N. F.
EXERCISE I.— AROMATIC WATERS
i. Aqua Cinnamomi. — In a dry mortar triturate i drop of cinnamon oil
with about 0.5 gm. of talc; then add, gradually and with continued tritura-
tion, 25 c.c. of water. Pass repeatedly through a filter until the filtrate is
perfectly clear.
Optional Preparations. — 2. By filtration similar to the cinnamon: Aq. Camphorae,
Peppermint.
3. By simple solution: Aq. Chloroformi, Creosoti.
4. By distillation : Aq. Anisi.
QUESTIONS
(a) Define the "aromatic waters."
(b) What is the object of the talc?
(c) What other methods could be used for making aromatic waters?
EXERCISE II.— LIQUORS
i. Liquor Calcis.— Slake 3 gm. of quicklime in an evaporating dish by
the gradual addition of 100 c.c. of distilled water. Stir occasionally during
half an hour. Let settle; decant the supernatant fluid and reject it . K
tin- insoluble residue into a bottle with 900 c.c. of distilled water; -
thoroughly; let stand twenty-four hours or longer. Shake again; let the
coarser particles subside and pour the fine suspension into another bottle.
Let stand, and pour off the clear "lime-water" as needed.
Optional Preparations.— 2. By simple solution: Liq. lodi Co.; Ac. Arscn.; Dobell's
Solution; Hypodermic Injections; Am{xnils.
mi. processes: Liq. Ammon. Acet; Chlori Co.; Fern Chlor.; Magn
Plumbi Subacet.; Potas. Arsenit.
QUESTIONS
(a) Define a "liquor."
(b) Explain the steps of the process for lime-water.
(c) Why is it necessary to use distilled water?
EXERCISE III.— SYRUPS, ELIXIRS, GLYCERITES, MUCILAGES
i. Sympus.— Heat 42.5 gm. of granulated sugar with 22.5 c.c
until di>M)lve.l. bofl; strain through dnth, adding through the strainer suf-
•er to make 50 c.c. (when cold).
5. if— Cinnamo
5. M.— Quicklime in 3 gm. portions.
42 A LABORATORY GUIDE IN PHARMACOLOGY
Optional Preparations.— 2. Prepared by adding the medicinal substance to syrup:
repared by dissolving sugar in the medicinal liquid: Syr. Fern lod.; Picis Liq.;
Pruni \
4. Elixir Aromaticum.
5. Glyceritum Boroglycerini.
6. Mucilago Acacia.
QUESTIONS
(a) Define * 'syrups/'
(b) How do they differ from "elixirs'? (c) From "glycerites"? (d)
From "mucilai:
(e) Why is the syrup boiled?
EXERCISE IV.— SPIRITS, COLLODIA
i. Spiritus Menthae Piperitae.— In a bottle dissolve i c.c. of oil of pep-
permint in 9 c.c. of alcohol; add o.i gm. of peppermint herb; macerate for
twenty-four hours or longer, and filter.
Optional Preparations. — 2. Spirits by simple solution: Spir. Ammon. Arom.; Camphor.
3. Collodia : Simple and Flexible.
QUESTIONS
(a) Define "spirits."
(b) How do they differ from "waters"?
(c) How from "tinctures"?
(d) What is the object of the peppermint herb?
(e) What is a "collodion"?
(f) How is collodion made flexible?
(g) Under what circumstances would simple and flexible collodion be
employed?
EXERCISE V.— INFUSIONS AND DECOCTIONS
i. Infusum Digitalis. — Crush 1.5 gm. of digitalis leaves in a mortar.
Pour on to this 50 c.c. of boiling water. Let stand one hour. Strain
through cloth. Add 15 c.c. of cinnamon water; and, through strainer, cold
water sufficient to make 100 c.c.
Optional Preparations. — 2. Barley Water: Wash i ounce of pearl barley; boil for
short time with \ pint of water. Decant and throw out the liquid. Add to residue 4 pints
of boiling water, boil down to 2 pints and strain.
QUESTIONS
(a) Define "infusions" and "decoctions."
(b) How do they differ from "solutions"?
(c) From "tinctures"?
(d) What is their strength when not specified?
(e) What is the strength of infusion of digitalis?
EXERCISE VI.— TINCTURES, FLUTOEXTRACTS, SOLID EXTRACTS, OLEO-
RESINS, RESINS
i. Tinctura Arnicae (by Maceration). — Crush 10 gm. of arnica in a mor-
tar. Transfer to a flask. Add 25 c.c. of official dilute alcohol (equal vol-
umes of alcohol and water). Cork the flask and shake.
5. M. — Peppermint oil; peppermint herb in o.i-gm. portions.
S. M. — Digitalis in i.s-gm. portions; cinnamon water.
5. M . — Arnica in lo-gm. portions; cinchona, powdered, in 2o-gm. portions.
CHAP. II
PHARMACEUTIC PREPARATIONS AND DISPI
43
Fig. i.— Method of percolation
(Thornton).
After a week strain through cloth and express strongly. To the residue
again add 25 c.c. of dilute alcohol, let stand a week and express. (Officially,
two portions of 12.5 c.c. a day apart.) Mix the liquids.
2. Tinctura Cinchonae (by Percolation). — Prepare a small percolator:
pack a little cotton loosely in the neck; over this pour an inch of sand
(Fig. i).
Mix 7.5 c.c. of glycerin with 67.5 c.c. of al-
cohol and 25 c.c. of water. In an evaporating
dish moisten 20 gm. of finely powdered cinchona
uniformly with 8 c.c. of this menstruum. Trans-
fer to the prepared percolator, without pressing.
Let it stand well covered for an hour (preferably
six hours).
Then pack it firmly (with the handle of the
spatula) and pour on enough of the menstruum to
saturate the powder and leave a stratum above
it. When the liquid begins to drop from the per-
colator close the lower orifice, and let the tightly
closed percolator macerate for forty-eight hours
(or until the next laboratory period).
Then let the percolation proceed slowly (about
10 drops per minute), pouring on the remainder of the menstruum, and
then enough of an alcohol-water mixture (67.5 A : 25 W, volume) until 100
c.c. of percolate are obtained.
Optional Preparations. — 3. Tinctures by dilution: Tr. Ferri Chlor.; Nuc. V'om.
4. Tinctures by maceration: Tr. Cardam. Co.
5. Tinctures by percolation : Tr. Aconiti; Digitalis; Gentian. Co.; Opii; Opii Deod.
6. Fluidextracts: Ergot; Glycyrrhiza; Rhubarb: Wild Cherry; Senna.
7. Solid extracts: Cascara Sagrada; Rhubarb; Gentian; Ergot.
8. Oleoresins: Capsicum.
0- Resins: Podophyllum.
QUESTIONS
(a) What are the advantages of maceration and percolation?
(b) Why is the maceration of tin arnica conducted in two or three stages?
(c) Why is it necessary to moisten the cinchona before placing it in the
percolator?
(d) Why is it necessary to allow the percolator to stand two days before
nin^ the percolation?
(e) Explain the differences between tinctures, fluidextracts, solid extracts,
oleoresins, and resin >.
EXERCISE VH.— MIXTURES
1. Mistura Cretae Co. — In a mortar mix prepared chalk (creta pr»-
parata), 3 gm.; acacia, a nm .: |><>\\<lered sugar, 5 gm. " (Thi- makes "com-
pound chalk powder.") Rub thi> mixture \\itli 4 e.r. of cinnamon v
and 2 c.c. of \\ater. gradually added, until a uniform mixture is obtained.
-iVr to a graduate and rinse the mortar with enough water to make
10 C.C.
2. Simple Suspension of Chalk. Rub prepared chalk with ;
c.c. of water. Compare the permanent e of this suspension with the pre-
ceding.
5. J/. — Cinnamon water.
44 LABORATORY GVIDK IN PHARMACOLOGY
3. Bismuth Mixture. — Make a mixture of bismuth subcarbonate, acacia,
cinnamon water, and water. There are to be 3 tablespoon doses; each dose
contain 0.5 iim. each of bismuth subcarbonate and of acacia, and equal
parts of cinnamon water and water.
Optional Preparation*.— 4. Lotio Nigra; Lot. Plumbi et Opii; Magma Magnesiae.
QUESTIONS
(a) Define a mixture.
(b) Why is it necessary to add acacia or sugar to suspensions of heavy
powders?
EXERCISE VIIL— EMULSIONS
i. Emulsum Olei Morrhuae (Official "Continental" Method). — In a dry
mortar rub 10 c.c. of cod-liver oil with 2.5 c.c. of finely powdered acacia
to a uniform smooth mixture. Then add at once 5 c.c. of water and trit-
urate lightly and rapidly until a thick homogeneous emulsion is produced.
To this add 2 c.c. of syrup and 3 c.c. of water. (The official emulsion is
flavored with 0.4 per cent, of wintergreen oil.)
Optional Preparations. — 2. Emuls. Asafet.; Emuls. Turpent.
3. Lecithin Emulsions. — Dissolve 5 gm. of Merck's lecithin in 50 c.c. of water and
triturate with 45 gm. of the oil (Bloor, 1013, Jour. Biol. Chem., 15, 112).
4. Casein Emulsion. — Raper, 1913, ibid., 14, 117.
QUESTIONS
(a) Define an emulsion.
(b) What are the proportions for making the "nucleus"?
(c) How does the gum act in helping the subdivision and suspension of
the oil?
(d) What other substances act as emulsifying agents?
(e) How can the thin (volatile) oils be emulsified?
(/) How are emulsions of gum-resins (asafetida) made?
EXERCISE IX.— LINIMENTS
i. Linimentum Calcis ("Carron Oil")- — Shake thoroughly 10 c.c. of
lime-water (calcium hydrate solution) and 10 c.c. of cotton-seed oil (offi-
cially, raw linseed oil) .
Optional Preparations. — 2. Liniments of Ammonia; Camphor; Chloroform; Turpen-
tine.
QUESTIONS
(a) Define a liniment.
(b) What are the usual bases of liniments?'
(c) What is formed in the preparation of the lime liniment?
EXERCISE X.— POWDERS
i. (Individual) Powder Papers.— Divide 10 gm. of starch into 10 pow-
ders, properly folded, as demonstrated.
Optional Preparations.— 2. Compound Powders: Compound Effervescent Powder;
Compound Licorice Powder.
3. Effervescent Salts: Effervescent Magnesium Sulphate.
4- Chemic Compounds: Saccharated Carbonate of Iron; Precipitated Sulphur.
5. if .— Cod-liver oil; syrup.
CHAP. II PHARMACEUTIC PREPARATIONS AND DISPENSING 45
QUESTIONS
(a) Why should compound powders be triturated in a definite order,
proceeding from the ingredient of the smallest to that of the largest bulk?
(b) What physical properties render a substance unsuitable for powder
papers?
(c) Why is it inadvisable to dispense a dose of less than 5 grains alone in
powders? How could this difficulty be overcome?
EXERCISE XI.— PILLS
i. (Individual) Glycyrrhiza Pills. — Place 2 gm. of powdered glycyrrhiza
in a mortar and incorporate excipient (glycerite of acacia) a little at a time,
sufficient to form a plastic mass. Care must be used lest too much excipient
is added (this may be remedied by adding a little dry acacia). The mass
must be worked very thoroughly, until it can be rolled in the hand without
breaking.
Dust the pill tile with a little starch, and on it roll out the mass to a
uniform cylinder extending over 10 or 20 divisions. Dust the spatula with
starch, and with it divide the cylinder into 10 exactly equal parts. Roll
each part to a spherical pill between the thumb and first two fingers. Place
the pills into the lid of the pill box, with a little starch, and roll them per-
feclly round with the ball of the thumb.
Pills must be of uniform size, smooth shape, and sufficiently firm to
resist gentle pressure. Several trials should be made if necessary.
Optional Preparations. — 2. Pills of Aloes; Ferrous Carbonate; Silver Nitrate.
3. Tablet Triturates; Compressed Tablets; Lozenges; Suppositories.
QUESTIONS
(a) What are the functions of the excipient?
(b) What other excipients are used in pills?
(c) What are the drawbacks of pills?
(d) What classes of substances should not be prescribed as pills?
(e) What are the ordinary limits to the size of pills?
EXERCISE XH.— CAPSULES
i. (Individual) Starch Capsules. — Divide 2 gm. of starch into 10 parts,
and place in capsules ("No. 3")* as demonstrated. Roll the finished cap-
sules in the hand (or better, clean cloth) to remove adherent powder.
Optional Preparations. — 2. "Soft" capsules of castor oil.
3. "Ampouls" (Proc. Amer. Pharrnaccut. Assoc., 57, 53).
QUESTIONS
(a) What are the advantages and disadvantages of capsules as compared
with pill>?
(b) What are the ordinary limits to the weight of capsules?
EXERCISE Xin.— OINTMENTS
i. (Individual) Unguentum Zinci Oxidum. — In a dry mortar rub 2 gm.
DC oxid with 10 gm. of benzoinated lard, gradually added, until they arc
5. U.— Powdered glycyrrhiza in a-gm. portions.
S.M. NO.JCUMK
5. J/.— Zinc oxid in a-gm. portion*; benzoinated lard in lo-gm. portions.
46 A LABORATORY GUIDE IN PHARMACOLOGY
thoroughly mixed and free from lumps. Or they may be mixed in a pill
tile; the zinc being placed at one side, the lard on the other, and the two
being gradually worked together in the middle by a spatula.
Optional Preparation*.— 2. Simple ointment; ointments of phenol; boric acid; sulphur;
tar; yellow mercuric oxid.
QUESTIONS
(a) Why is it essential that the ointment be free from lumps?
(b) What are the relative advantages and disadvantages of lard, petro-
latum, and wool-fat?
EXERCISE XTV.— POULTICES
i. Cataplasma Lini. — Boil 50 c.c. of water and a small pinch of sodium
bicarbonate; stir into this gradually ground flaxseed (linum contusum)
until a thick mush is obtained (about 50 gm.).
Optional Preparation. — 2. Spreading a plaster.
QUESTIONS
(a) Why is the linseed added to the water and not vice versa?
(b) Why is the soda added?
CHAPTER III
INCOMPATIBILITY
It is assumed that the student, through analytic chemistry, is familiar
with most of the reactions which underlie incompatibility. Only a few of
these are reviewed here as types. They should be performed by each
student individually. The criticism of the incompatible prescriptions will
need some aid from the instructor. They may be compounded as optional
experiments.1
EXERCISE I.— EXPLOSIVES
1. (Optional) Potassium Chlorate. — Rub a trace of the chlorate and tannin in a
mortar: detonation.
2. (Optional) Nitric Acid. — Mix some strong nitric acid and alcohol in a beaker, and
let stand under a bell jar: in a short time orange vapors arise, and suddenly the solution
boils up and is thrown from the beaker.
3. Chromates. — Kr2Cr2O7 solution -f Alcohol: no change. (There
may be a slight precipitate, which redissolves if a little water is added.)
Add concentrated H2SO4. Green color, and evolution of gas.
[K2Cr2O7 + H2SO4 = 2CrO3 + 2KHSO4 -f- H2O
Cr03 + 3C2H60 = Cr203 + 3C2H4O (aldehyd) + 3H2O
3H2S04 = Cr2(S04)3 + 3H2O]
5. If .—Ground flaxseed.
1 An extensive compilation of incompatibilities is contained in Ruddiman's "Incompatibilities
in Prescriptions," New York, 1908.
CHAP. Ill INCOMPATIBILITY 47
QUESTIONS
(a) What other substances would explode when rubbed with tannin?
(b) With chlorate?
(c) In what order should the ingredients of the following gargle be mixed
to avoid explosion?
If. KC1O3 0.5 1 If put up without heating, no change
Aquae . . 10.0 ! will occur, illustrating the ppssibil-
Glycerini 2.0 j ity of mixing certain explosives in
Tr. Ferri Chlor i.o J solution.
(d) In what order should they be mixed if the gargle is to contain free
chlorin?
(e) What salts oxidize organic matter even in dilute solution?
CRITICIZE THE FOLLOWING PRESCRIPTIONS
(a) Potas. Nitrate. (6) Silver Nitrate. (c) Tr. lodin.
Sulphur. Cocain. Ammon. Chlorid.
Water. \\ uter.
(d) Potas. Permang. (e) Silver Nitrate.
Glycerin. Tap Water.
Water.
EXERCISE H.— IODLDS
1. Liberation of lodin by Oxidizing Agents. — Mix solutions of potassium
iodid and hydrogen peroxid: brown color. (2KI -f H2O2 = 2KOH -f 2!.)
2. Precipitation of Metallic Salts. — (a) Mix solutions of potassium iodid
and lead acetate: yellow precipitate.
(b) Mix solutions of potassium iodid and mercuric chlorid: red precipi-
tate, soluble in excess of either reagent.
(c) Mix solution of potassium iodid and a little calomel: yellow color
(mercurous iodid), gradually changing to green, gray or black, by decompo-
sition into metallic mercury and mercuric-potassium iodid.
3. Precipitation of Strychinn.— To 10 c.c. of KI solution (5 per cent.)
add 10 drops of i per cent, strychnin sulphate. Keep until crystals of strych-
nin iodid develop (if necessary until next laboratory period).
QUESTIONS
(a) What other substances evolve I from KI?
(b) What other metals precipitate with iodids?
(c) What metals are precipitated by bromids?
(d) By chlorids?
(e) Explain the changes with KI and H.
(/) Why is it dangerous to administer calomel to a patient receiving
iodids?
(R) Are most alkaloids precipitated by KI?
Ky HHr?
CRITICIZE THE FOLLOWING PRESCRIPTIONS
\al. (6) Tr. lodin. (c) KI
i.tlu-rNitr. Potas. Permang. I.i«i Potas. Am
Water. Water. Water.
(d) Silver Nitrate.
Normal Salim-.
S. If.— Strychnin sulphate i per cent.
48 A LABORATORY GUIDE IN PHARMACOLOGY
EXERCISE III.— ALKALIES
i. Precipitation of Earths. — (a) Mix solutions of magnesium sulphate
and sodium carbonate: white precipitate of magnesium carbonate.
(b) Mix solutions of magnesium sulphate and sodium bicarbonate: no
precipitate
a. Precipitation of Metals.— (a) Mix solution of sodium bicarbonate and
tincture of ferric chlorid: evolution of CO2 and precipitation of brown ferric
.carbonate.
(b) Mix solutions of ferric ammonium citrate and sodium bicarbonate:
no precipitate.
(<•) Mix solutions of alum and sodium borate: white precipitate of
aluminum hydroxid.
(d) Mix solutions of alum and boric acid: no precipitate.
3. Precipitation of Alkaloids. — (a) Mix liq. potas. arsenitis with a few
drops of saturated quinin sulphate: precipitate of quinin. Add a few drops
of dilute hydrochloric acid: solution.
4. Decomposition of Chloral. — Mix solutions of chloral and sodium hy-
droxid: odor of chloroform. (CC13COH + NaOH = NaCHO2 + CHC13.)
5. Decomposition of Hexamethylenamin by Acids. — To a solution of
hexamethylenamin add dilute HCl and heat: odor of formaldehyd. (CH2)6-
N4 + 6H20 = 4NH3 + CH20.
QUESTIONS
(a) Are all salts of earths precipitated by alkalies?
(b) Why does not the bicarbonate precipitate the magnesium?
(c) Does it also prevent the precipitation of the metals?
(d) Are the salts of all metals precipitated by alkalies?
(e) Why is the double citrate not precipitated?
(/) Would the sodium borate precipitate alkaloids?
(g) How could the precipitation of alkaloids by the arsenite be pre-
vented?
(h) Which salts act as alkalies?
CRITICIZE THE FOLLOWING PRESCRIPTIONS
(a) Liq. Calcis. (b) Magn. Sulph. (c) Tr. Nuc. Vom.
Sod. Bicarb. Sod. Phosph. Sp. Ammon. Arom.
Elix. Arom.
(d) Bism. Subnitr. (e) Syr. Scillae. (/) Hexamethylenamin.
Sod. Bicarb. Ammon. Carb. Ammonium Chlorid.
Water.
EXERCISE IV.— SALICYLATES
1. Precipitation by Acids. — Mix solution of sodium salicylate with dilute
hydrochloric acid: white precipitate of salicylic acid.
2. Color with Iron.— To a solution of sodium salicylate add a few drops
of ferric chlorid: violet color of ferric salicylate.
3. Precipitation of Quinin. — Mix solutions of quinin sulphate (saturated)
and sodium salicylate: white precipitate of quinin salicylate.
QUESTIONS
(a) Why is sodium salicylate usually given with sodium bicarbonate?
(b) What other substances give colored solutions with iron?
(c) Do most alkaloids precipitate with salicylates?
CHAP. IV ISOLATION OF POISONS 49
CRITICIZE THE FOLLOWING PRESCRIPTIONS
(a) KI. (b) Sp. jEth. Nitr. (c) Sod. Salic.
Sod. Salicyl. Sod. Salir. Antipyrin.
Water. \\";iter. Make a powder.
EXERCISE V.— TANNIN
1. Precipitation of Metals. — Mix solutions of tannin and mercuric
chloric! : white precipitate of mercuric tannate.
2. Precipitation of Alkaloids. — Mix solutions of quinin sulphate (satu-
rated) and tannin: gray precipitate of quinin tannate. Add alcohol: solu-
tion.
QUESTIONS
(a) Are all metals precipitated by tannin?
(b) What other change occurs with ferric salts?
(c) Are all alkaloids precipitated by tannin?
(d) Are glucosids precipitated by tannin?
(e) What other substances are precipitated by tannin?
CRITICIZE THE FOLLOWING PRESCRIPTIONS
(a) Tr. Ferri Chlor. (6) Liq. Potas. Arsen. (c) Ac. Tann.
Tr. Cinchon. Ac. Tann. 1 1 < ),-.
Water. Water.
(d) Gelatin.
Tannin.
Water.
EXERCISE VI.— PHARMACEUTIC INCOMPATIBILITY
1. Alcoholic Preparations and Water. — (a) Mix Sp. Ammon. Arom. and
water: precipitate of oils.
(b) Mix Tr. Myrrh and water: precipitate of the resins.
2. Alcohol and Water-soluble Drugs.— (a) Mix Muc. Acacia with akro-
hol: precipitate of the gum.
(b) Mix Sat. Sol. Sod. Chlorid with alcohol: precipitate of the salt.
Add water: solution.
5. Solubility.— Mix i part liquefied phenol with 10 parts ol \\ukr. How
can this be brought into solution?
QUESTIONS
(a) Are all alcoholic preparations incompatible with water?
(b) Why is the salt precipitated by the alcohol?
CHAPTER IV
ISOLATION OF POISONS
The >ons by the students themselves requires more time than can be
usual 1 course. It would also require very
their tit ally trustworthy. isi> of this « hap;
presented A good presentation of 'I <n in
Gadat rl)u. h <ler . IK-HUM hen Tozkobgfe," Gocttingcn, 1909; in Abdcrhalden's
Handb., 5, 673; tenricth, 1915.
S. M— Sp. ammon. arom.; tr. myrrh*; mucil. acaci*.
5°
A LABORATORY GUIDE IN PHARMACOLOGY
VOLATILE POISONS
EXERCISE I.— (DEMONSTRATION)
A mixture of meat or similar material, poisoned with phenol (or hydro-
cyanic acid, chloral, alcohol, etc.), is diluted with water, acidulated with
tartaric acid, and distilled from a flask through a Liebig condenser. The
distillate has the characteristic odor of the substance and may be subjected
to the corresponding tests.
EXERCISE II.— (DEMONSTRATION) DISTILLATION TEST FOR PHOS-
PHORUS
The poisoned material is placed in a flask connected with a steam kettle
and vertically descending Liebig condenser (Fig. 2) arranged in a dark
room. The air is expelled from the flask by steam ; the flask is then heated.
The characteristic luminous ring appears in the tubes or condenser, shifting
Fig. 2. — Mitscherlich apparatus.
its position according to the heat applied,
substances interferes with the test.
The presence of other volatile
EXERCISE m.— (DEMONSTRATION) ISOLATION OF FIXED ORGANIC
POISONS BY MODIFIED STAS-OTTO METHOD
The instructor should perform the experiment in advance, preparing
the various stages of the separation; so that only the steps of the process
need be demonstrated, without halting the demonstration to wait for the
separation to take place.
1. Extraction. — To a mixture of 30 gm. of hashed meat and 3 gm. of
powdered nux vomica add about 100 c.c. of water and a pinch of tartaric
acid. Boil for ten minutes. Cool. Strain through Canton flannel. Reject
the solid residue.
2. Removal of Salts, Proteins, and Fats. — Add about 10 gm. of sand (or
some purified oak saw-dust) to the strained solution, and evaporate, first on
free flame, then on water-bath, to a paste. Add 40 c.c. of 95 per cent, alco-
hol, let stand ten minutes or longer, with frequent stirring; and filter. The
salts, proteins, and fats are left on the filter, since they are insoluble in
alcohol. These are rejected. The alcoholic solution contains the organic
poisons.
3. Removal of Resins, Fats, etc. — Dilute the alcoholic solution with an
CHAP. IV
ISOLATION OF POISONS
men
51
equal volume of water. This precipitates the above impi
resins and croton oil would be found in this precipitate) . Filte
precipitate. Evaporate to near dryness to remove the alco
the residue in 50 c.c. of water. Filter. Assure yourself that
acid.
4. Removal of Neutral Principles and Some Other Impt
rities
. RAjtoJttersity
ol. Dissolxf
the filtrate is
1 oronto
Tftifg Place
the solution in a separating funnel, add 25 c.c. of ether, and shake with a
gentle rotatory motion for ten minutes. Separate the two layers. The
ethereal layer would contain the neutral principles, which could be obtained
by evaporating the ether. In the present instance the ethereal layer is
rejected. The watery layer contains the alkaloidal salts. It is treated by
($)•
5. Extraction of Alkaloids.— Replace the watery solution of 4 in the sep-
arating funnel. Add ammonia until it is freely alkaline (this liberates the
free alkaloids, which are soluble in ether. The alkaloidal salts are insoluble
and were, therefore, not extracted in 4). Add 25 c.c. of ether and shake with
a rotatory motion for ten minutes. Let the liquid separate, and draw off
the watery layer (which would contain morphin); this is rejected. The
ethereal layer contains most of the alkaloids. Distil off the ether. Test
some of the residue for Strychnin and Brucin. Dissolve another portion in
a little dilute sulphuric acid, inject into a frog, and note the convulsions.
(The ether extractions would be repeated, in practice, as long as they would
take up any alkaloid.)
Explanatory Note.— The method rests on the different solubility of the constituents
of the mass in successive solvents. It may be represented diagramatically as fell
Extraction with boiling dilute tartaric acid.
Solution:
Evaporation and extraction
with alcohol,
Residue:
Coagulated
Protein, Fiber,
Solution: Evaporation of alcohol and ex-
tra it inn with water,
Residue: Salts, proteins, fats.
Extraction of acid solution with ether,
Residue: Resins, fats, volatile oil>. croton
oil, » hlorophyll, etc.
Addition of ammonia to watery solution,
liberation of alkaloid, extraction with
ether.
Kiln-real Layer: Neutral principles, gluco-
. cantharidin, caffcin, and some
other alka!
Acidulntion, alkalinisation with ammonia,
extraction with an-ii. ether, chloroform,
or hot amyl aliohol.
Ethereal Layer: Bulk of alkaloids.
Watery Layer: Inorganic Poisons.
Ethereal Layer: Morphin.
TECHNICAL NOTES
| is ,,1'trn a \rry .li-turl.in^ - It is less
liable to,,, , nr r !.»nr with a very (fffj
IV I. a Wall. 1014, advocates a special type
innrl to avoid rimil-iri. atr IMiarm. Assoc., 3, 498). The
theory ><lc solvents is discussed by Gadamer, 362.
52 A LABORATORY GUIDE IN PHARMACOLOGY
EXERCISE IV.— (DEMONSTRATION) DESTRUCTION OF ORGANIC MATTER
FOR ISOLATION OF INORGANIC POISONS BY FRESENIUS-BABO
METHOD
Organic matter more or less obscures the reactions of inorganic poisons
and must, therefore, be destroyed. The Fresenius-Babo method1 is gener-
ally preferred.
(A) Place the material (meat, etc., poisoned with arsenic) in a liter flask
with as much arsenic-free HC1 as would correspond to the dry material.
Dilute with sufficient water to make a thin gruel. This is heated luke-warm
on a water-bath, and potassium chlorate added at intervals, 0.5 gm. at a
time, until the material is practically dissolved (not more than 4 to 6 gm.
should be used). The solution is then boiled in an evaporating dish to 100
c.c., or until free from chlorin, diluted to 400 c.c., 2 c.c. of dilute sulphuric
acid are added, and the mixture is set aside over night, and filtered. The
filtrate (B) would contain most of the metals; the residue (K) would con-
tain Ag, Ba, and Pb. The further separation is effected by the schema given
below. Only the test for arsenic need be demonstrated.
Marsh's Test (see B). — Produce hydrogen in flask by acting on pure
zinc with arsenic-free HC1; pass through CaCl2, then through tubes drawn
Fig. 3. — Marsh apparatus.
out at several places (Fig. 3). Heat to redness at the thick portion of a
segment. (This blank test should be continued for six hours.) If no mirror
appears, introduce the suspected solution. Black mirror occurs with arsenic
or antimony. They may be distinguished as follows:
ARSENIC:
Mirror beyond heated portion.
Garlic odor on heating in air.
Dissolves in hypochlorite.
Easily volatilized when heated in
hydrogen.
Heated in air, yields easily vola-
tilized white crystals.
Heated in H2S, yellow, insoluble
ble in HC1.
Dissolved in HNO3, evaporated,
plus AgNO3, plus vapor of NH3, red
or yellow precipitate.
ANTIMONY:
Mirror at head portion.
No odor.
Not.
Not easily volatilized.
Amorphous white residue, not easily
volatilized.
Red (black on strong heating);
soluble in HC1.
No color in cold; black (metallic Ag)
on heating.
Further details, Gadamer, no.
CHAP. IV ISOLATION OF POISONS 53
SCHEMA FOR ISOLATION OF METALS
Filtrate B. — Pass through filter water to just 500 c.c. Use 50 c.c. for Marsh's test.
If As is present, use remainder for quantitative (see C). If not, evaporate small sample,
dissolve in 10 c.c. water, add NH<OH: blue = Cu.
C. — Heat remainder of filtrate B to 80° C. and pass arsenic-free H2S for two or three
hours, until cool. Heat again, and repeat. Stopper and set aside in warm place for
twenty-four hours. Precipitate may contain As, Sb, Hg, Cu, Pb. It may be used for
the quantitative estimation of As, or for further identification by D. Filtrate = I.
D (H*S precipitate of C), — Wash with HjS water, warm with 4 c.c. ammon. sulphid,
4 c.c. ammonia, 8 c.c. water. Filter. Filtrate = E; Insoluble = F.
E i'iltrate of D). — Evaporate to dry; heat with HNOj until pure yellow; heat to expel
1 1 \< )3; add NaiCO, and NaNOj; fuse; extract with boiling water; add 2 gm. XaHCO»;
filter; Filtrate contains As and may be used for quantitative. The insoluble = Sb: apply
late.
F Insoluble of D). — Oxidize residue and filter in capsule with HC1 and KClOs; filter;
dilute; heat; pass H2S; filter; wash precipitate with warm HNOS. Filtrate = G. Pre-
cipitate = II.
G (Filtrate ofF). — Add 10 drops dilute HjSO<; evaporate; take up with water. Residue
= Pb: Filtrate = Cu. (Apply tests.)
H (Precipitate of F).— -Oxidize with aqua regia; evaporate; filter; dilute; test for Hg.
I (Filtrate of C). — Use half for zinc, half for chromium.
Zn: Neutralize with KOH; acidulate with H,CiO?; precipitate with H,S; wash
precipitate with HjCsOj in H2S water (i : 5); incinerate, precipitate, and filter;
dissolve in dilute H2SO4, plus a little HNOs; evaporate dry; dissolve in HjO;
test for Zn.
(.>: Kvaporate to just moist; mix with KNO»; dry; fuse; dissolve; test for chromate.
K (Residue of A).— Fuse with KXO,, Na2CO,, and NH^O,. Suspend in H,O;
pass CO2; boil; filter. Dissolve precipitate in dilute HNOj. Test this solution for Ag,
Ba, and Pb.
Electrolytic Determination of Metals. — Directions are given in the U. S. P. DC and
in Gadamer, 130.
EXERCISE V.— (OPTIONAL) ALKALOIDAL ASSAY
The U. S. P. process for Belladonna is typical of the majority of assays (with the im-
portant exception of opium). It consists in a modification of Keller's method. The
quantitative estimation of alkaloids is also described in Gadamer, 496; Abdcrhalden's
Handb., 6, 120; Autenrieth (Warren, 1915), pp. 86 and 246; and in the monograph of
von Korczynski, "Methoden der exacten quantitativen Bestimning der Alkaloide,"
Berlin, 1913.
EXERCISE VI.— (OPTIONAL) PHARMACEUTIC TESTING
The U. S. P. tests for purity are well illustrated by the following:
1. Sodium Bromid.
2. Time limit test for heavy metals.
I ron Sulphate.
4. Acrtphrnetidin.
1 ilnin Sulphate.
6. Chloroform.
ther.
TECHNICAL REFERENCES
•'-'on of Melting Point.— U.S. P. IX; Abderhalden's Handb.. i. 208; Menge,
1910, Hyg. Lab. Bui. No. 70.
\bderhalden, i, 214; Small quantities, Gadarm
for moloular wriirht. Abderhaldrn. 6, 364.
,///>• Df termination— -V. S. P. IX; Abderhalden, i, 451; Scidcll, 191 c
Lab. Bui. No. 67.
QUESTIONS ON CHAPTER IV
(a) Why i- it necessary to acidulate the material before the distillation
olatilc poisons?
(b) Why i- an organic acid used rather than a mineral a-
(c) Why is it advantageous to conduct the distillation with live steam?
54 A LABORATORY GUIDE IN PHARMACOLOGY
(d) In testing for phosphorus, why must the air be expelled from the
flask before heating?
(e) Does the failure of the luminous ring-test exclude phosphorus?
(/) What is the general principle of the extraction of fixed organic
poisons?
Of) Why is the material boiled with acidulated water?
Sh) Would this be necessary when working on urines?
i) How would the process be modified in this case?
(j) How would it be modified if -the suspected poison is difficultly soluble
iter?
(k) In the ethereal extraction, would it make any difference whether
30 c.c. of ether is used at one time, or in three 10 c.c. fractions?
(/) Why is it necessary to destroy the organic matter when searching
for mineral poisons?
(m) How is this accomplished?
(n) Why would it be inadvisable simply to incinerate the material?
(0) State the principle of the Marsh test.
CHAPTER V
SPECIAL TESTS OF IMPORTANT ALKALOIDS
The main object of these exercises is to familiarize the student with the
reactions which are utilized in toxicologic analysis and in the urine, food, etc.
It must be remembered that impure products give these tests very imper-
fectly. They may, however, be applied to tablets,1 capsules, etc., especially
if these are first extracted with suitable solvents. When the dry substance
is used, the reaction is performed on a glass slide or watch-glass, placed on
white paper; or on a porcelain slab. A piece of broken evaporating dish
may be used if the reaction requires heat. A mere trace of the substance,
about a milligram, should be employed. When solutions are used, the
reactions are generally made in a test-tube or capsule. The student should
remember that he is handling very strong poisons.
The tests need not be memorized, but should be described in the notes
or checked in the book. Two students may work together. The physiologic
tests are stated for convenience of reference, but need not be performed at
this time.
EXERCISE I.— STRYCHNIN
(Physiologic test: peculiar convulsions in frogs or mice.)
To a trace of the powdered alkaloid add:
1. A drop of concentrated H2SO4: no change; then a small crystal
K2Cr2O7. Play of colors through blue, violet, red, orange (Otto).
2. A drop of concentrated NH03; heat gently: with most samples a yel-
low color, due to Brucin.
3. Determine the dilution at which the bitter taste of strychnin just dis-
appears (begin with i : 50,000; to 5 c.c. of this add water in portions of
i c.c.). Report your results. (The usual limit is i : 40,000 to i : 67,000.
Should any student depart markedly from this, he should try his sensitive-
ness to other bitter substances,)
> Analysis of Tablets, Kebler, 1914, Jour. Amer. Pharm. Assoc., 3, 6. 107.
CHAP. V SPECIAL TESTS OF IMPORTANT ALKALOIDS 55
4. (Optional). — Strychnin, even in very dilute solutions, gives a white precipitate with
chlorin water.
5. (Optional) Isolation of Strychnin. — Proceed as in Chapter IV, Exercise III, but
in (5) use chloroform as solvent.
6. (Optional) Picronolic Acid for Purification of Strychnin and Other Alkaloids. —
This was proposed by \Y. II. Warren and Weiss (Jour. Biol. Chem., 3, 330, 1907). The
picronolate of strychnin being very insoluble, may be precipitated from aqueous solution,
thus separating it from other substances that interfere with purification.
7. (Optional) Quantitative Determination. — Salant, 1904, Jour. Med. Res., 12, 51.
8. (Optional) Determination of Strychnin in Tablets. — Kebler, 1914, Jour. Amer.
Pharni. Assoc., 3, 1098.
9. (Optional) Brucin. — This is important mainly because of its association with
strychnin in nux vomit a.
(a) To a little of the powdered alkaloid add a small drop of nitric acid: blood-red color.
Add a few drops of i per cent, sodium thiosulphate (hyposulphite): violet color (Cotton).
(b) To some powdered Nux \ <>mua add a drop of concentrated HNOi; orange color,
due to Brucin.
EXERCISE II.— CAFFEIN
1. Moisten some powdered alkaloid with nitric acid: yellow to orange
color. Evaporate the excess of acid on water-bath and expose to ammonia
vapor: garnet to purple color (murexid reaction of Stenhouse, Rochleder).
(Theobromin and theophyllin give very similar reactions.)
2. (Optional) Isolation of Methylxanthins.— Proceed as in Chapter IV, Exercise III,
i to 4. Then extract the ac id solution with chloroform, which dissolves the methylxanthins.
3. (Optional) Isolation from Urine. — The acid urine is shaken directly with chloro-
form, which dissolves the methylxanthins, but not the normal urinary xanthin bases.
TECHNICAL REFERENCES
Quantitative Estimation. — Off. Agric. Chem., Abderhalden's Handb., 2, 610; 6, 132;
Preparation. Abderhalden, 2, 959.
Coffee, Tea, and Chocolate, ibid., 7, 373; Detection of Chicory in Coffee decoctions,
A all, Amer. Jour. Pharm., 85, 535.
Theobromin and Thcophyllin, Preparation and tests, Abderhalden's Handb., 2, 610,
060.
EXERCISE HI.— MORPHIN
i. To a solution (about i : 1000) of morphin sulphate add a little fresh
sodium iodate solution, a few drops of dilute sulphuric acid, and a little
starch-paste: purple color. This is a very delicate test, but is also given by
other reducing substances (Mohr).
!'«> a little (2 per cent.) aqueous solution in a test-tube add a drop of
(neutral) ferric chlorid: blue color (Schaer); not delicate.
To a trace of powdered alkaloid add a drop of nitric acid and heat:
ure color.
I o a trace of dry alkaloid add a drop of fresh Marquis' (Kobert's)
cnt (concentrated H2SO4, 20 c.c.; 40 per cent, formalin, i c.r.). Play
lots from purplr-ml t<» viulrt blue.1
5. Mix a trace of dry alkaloid with an equal quantity of ammonium
molybdate, and add a drop of concentrated sulphuric acid (Froehde's
reagent): violet color, chan^in^ to deep blue.
S. M.— Strychnin sulphate solution, i : 50,000.
5. J/.— Starch paste; Marquis* reagent; ammon. molybdate; \ gr. morphin tablets.
it ximil.ir reactions with phenols and t! ive» (carbolic
i 'ir (..l..r u: • b, however,
.nil murphin. . .irl».li, acid being the only on- ild give rise to a
take. This can be removed by boiling the acidulated solution until it ceases to give the phenol
reactions (Hatcher).
56 A LABORATORY GUIDE IN PHARMACOLOGY
6. To a few drops of (2 per cent.) aqueous solution in a test-tube add
about 2 c.c. of concentrated HC1 and a few drops of concentrated H2SO4.
Boil in water-bath for one-half hour: apomorphin is formed. Neutralize
i Na,CO3 (solution) and add a drop of Tr. lodin: emerald color. Shake
with ether: this takes a violet color (Pellagri's reaction— also given by codein,
heroin, etc.)-
7. Morphin in Tablets, etc.— (a) Dissolve J-grain tablet in a few drops
ater, and apply Tests i and 2.
(b) Crush another tablet, shake with chloroform and a drop of am-
monia; filter; evaporate on three watch-glasses; apply Tests 4 and 5.
8. (Optional) Quantitative Estimation in Tablets. — Rep. Chem. Lab. Amer. Med.
Assoc., 1913, 6, 88 (precipitation by ammonia); Kebler, 1914, Jour. Amer. Pharm. Assoc.,
3, 1093; in Tablets and Pills, H. \V. Jones, 1915, Jour. Amer. Pharm. Assoc.
9. (Optional) Isolation of Morphin from Tissues, etc. — Proceed as in Chapter IV,
Exercise III, using chloroform or amyl alcohol in No. 5.
10. (Optional) Quantitative Isolation. — Ruebsamen, 1910, Arch. exp. Path. Pharm.,
64, 54; Kaufmann-Asser, 1913, Bioch. Zs., 54, 161.
Physiologic Test. — Erection of mouse-tail.
TECHNICAL REFERENCES
Further tests for Morphin. — T. H. Oliver, 1914, ref., Jour. Amer. Med. Assoc., 63, 513.
Isolation from Tissues. — Gadamer, 551; G. L. Schaefer, 1913, Amer. Jour. Pharm.,
85, 4395 Cloetta, 1003, Arch. exp. Path. Pharm., 50, 455; Thorburn, 1911 (Phenyl ethyl
alcohol), Jour. Ind. Eng. Chem., 3, 754; Girard, Del&irde and Ricquet, Bioch. Centr.,
4, 451. Ptomains do not interfere, Rosenbloom, 1914, Jour. Biol. Chem., 18, 131.
Quantitative Estimation. — Abderhalden's Handb., 6, 126; Gadamer, 551; Sanger and
Broughton, 1909 (adaptation of Marquis' test), Proc. Soc. Biol. Chem., i, 250; Gordin
and Harrison, 1906 (in presence of glycerin), Jahrb. Pharm., 66, 308.
Preparation of Opium Alkaloids. — Abderhalden, 2, 942.
EXERCISE IV.— CODEIN, HEROIN, AND RELATED ALKALOIDS
Codein and heroin, as all esters of morphin, give the Pellagri reaction
(Exercise III, No. 6) for apomorphin. They do not give the reactions i and 2.
Special reactions are as follows:
1. (Optional) Codein. — Place a little of the dry alkaloid in a capsule and add a few
drops of concentrated HaSO^ faint greenish, then violet, color. Add a drop of concen-
trated HNO3: plays from yellow to purple.
2. (Optional) Heroin. — A little of the dry alkaloid is dissolved in a watch-glass in a
few drops of nitric acid: yellow color; on standing or heating this turns greenish blue
and then fades again to yellow.
3. (Optional) Acetyl Radicals of Heroin.— Heat a trace of heroin with dilute sul-
phuric acid in a test-tube, add some alcohol and boil: acetic odor.
4. (Optional) Dionin. — This gives most of the reactions of codein, but somewhat
different colors with Marquis' reagent.
5. (Optional) Isolation of Codein, Dionin, Peronin, and Heroin. — Codein, dionin,
and peronin are extracted from the alkaline solution by ether (Chapter IV, Exercise III),
using sodium carbonate in place of ammonia in No. 5. The extraction of heroin is as
for morphin.
6. (Optional) Narcotin. — To a trace of powdered alkaloid add some concentrated
sulphuric acid: greenish yellow solution, turning to orange, intensified on heating. On
continued heating, violet with purple streaks (Arnold).
7. (Optional) Papaverin. — L. E. Warren, 1915, Jour. Amer. Chem. Soc., 37, 2402.
8. Meconic Acid (Serving as a Test for Opium). — Dilute a few drops of
tinct. opii with water and add drop of ferric chloric! : red color, not bleached
by HgCl,.
S. M .— Tr. opii.
CHAP. V SPECIAL TESTS OF IMPORTANT ALKALOIDS 57
9. Apomorphin. — (a) To a trace of dry alkaloid add a drop of nitric acid:
blood-red color.
(b) To a few drops of (about) i : 500 watery solution (note the green
color) add 5 drops of Na2CO3 and a drop of alcoholic iodin: emerald color.
Shake with ether. This becomes violet.
(c) (Optional). — To 5 drops of apomorphin solution add 5 drops of saturated solution
of mercuric chlorid; then 5 drops of 10 per cent, sodium acetate. Boil for a few minutes,
cool, and add i to 2 c.c. of amyl alcohol. This is colored blue on shaking. The test is
extremely delicate (to i : 500,000, Amer. Jour. Pharm., 87, 564, 1915).
(d) (Optional). — Apply test (6) to apomorphin tablets.
(e) (Optional). — Presence of Apomorphin in Morphin. — To a dilute solution of the
hydrochloric! add 3 drops of i per cent, potassium ferrotyanid, and shake with benzol.
It apomorphin is present, the benzol acquires an amethyst color. On shaking with N'aOH,
this turns reddish violet, deepening to violet on standing (sensitive to 0.003 m8-» Feinberg,
1913, Zs. physiol. Chem., 84, 363).
10. (Optional) Hydrastin. — (a) Dissolve in dilute sulphuric acid and add dilute potas-
sium permanganate: blue fluorescence (hydrastinin).
(b) To the dry alkaloid add concentrated sulphuric acid: no color; heat: violet.
Isolation. — As per Chapter IV, Exeicise III, i to 5.
(d) Hydraslin in Fluidextract Hydraslis. — Shake 5 drops of the fluidextnut with
of 5 per cent, sodium bicarbonate and 10 c.c. of ether. Wash the decanted ether
layer with 5 c.c. of water. Filter the decanted ether layer and evaporate it to dryness.
Dissolve the residue in 10 c.c. of dilute sulphuric acid, and add 12 to 15 drops of i : 1000
potassium permanganate-. The solution is decolorized, but after dilution with 5 volumes
of water it shows a blue fluorescence in reflected light (Glueckmann, 1913, Pharm. Post.,
348).
11. (Optional) Berberin. — (a) Note yellow color of solutions, even when very dilute.
(6) To a solution add chlorin water: red color (Klunge).
(c) To a solution add solution of KI: precipitate.
(d) Quantitative. — Richter, W., 1914, Arch. Pharm., 252, 192; ref., Zentr. Bioch.
Bioph., 17, 476.
(e) Berberin in Fluidextract Hydraslis. — To 10 c.c. of concentrated hydrochlori
add a drop of the fluidextract, shake, add a drop of hydrogen dioxid solution, and shake
again: a persistent red color develops in five to ten minutes (Glueckmann, 1913, Pharm.
Post., 348).
TECHNICAL REFERENCES
:n Tests. — Zemick, 1903, Jahrb. Pharm., 339 — in excreta, Langer, i
0, 222; Rapid Dettrminalion of small quantities 1\. Miller. 1015. Amer. Jour. Pharm.,
;8. Separation of Heroin and Morphin, Doran, 1916, Jour. Amer. Pharm. Assoc.,
S» 163.
Tests for Minor Opium Alkaloids— \.. K. \Yurren. 1015, Amer. Jour. Pharm., 87, 437.
ApomorpMn Preparation.-- Abderhalden's Handb., 2,956; Pun
:: morphin. Reagent. Ho-hida, loo*. I". S. p. Diue-t, 353. Tests in presence of
morphin. (.rimhert ami I.e. ler. . 1014. ref.. Zentr. Bio<h. Bioph., 18,625.
' /J//5, Alkaloids, Preparation. — Abderhalden, 2, 945.
EXERCISE V.— COCAIN AND ANESTHETIC BASES
Physiologic Test for Cocain.— Local anesthesia and dilation of pupils.
1. (Optional) Cocain.— (a) To a solution of , .>. ain hydro, hlori.I ,.n a slide add tome
. h.ir.i. teri-ti. \ i"|. :
(b) Triturate s< with an equal (about) quantr M with
dilute alcohol: turn-* fray l.y re<lu< lion <>f mercury (Fluecklger). Dilute \\ith a little
and boil. I ruity odor of methyl benzo
(c) Add a crystal of cocain to solution of alpha naphthol in 40 per
(d) Isolation of Cocain— As per ( h.pt.t I\ i
:itati:r Determination. — Rifat \v:u hdani. i«
•ptional) Distinction of Cocain and Substitutes. Setter and Kngcr, i
Jour. Pharm., 8.}, IQ>; (larlamer, 576.
S. J/.— Alx,m.,rj,hi.» IK 1 i : 500 solution.
58 A LABORATORY GUIDE IN PHARMACOLOGY
TECHNICAL REFERENCES
Demonstration of Cocain— Vardan, 1908, Bioch. Central., 8, 169; Hankin, Jahrb.
Pharm., 71, 194.
Preparation.— Abderhaldcn's Handb., 2, 929.
EXERCISE VI.— ATROPIN AND RELATED ALKALOIDS
The following tests are given by all the solanaceous mydriatic alkaloids
and their derivatives:
1. Place a trace of dry atropin in a test-tube. Add 10 drops of con-
centrated HjSO4, and heat until it becomes brown; then add 2 volumes of
water: characteristic odor, resembling tuberose (Gulichno), strengthened
by KMnO4 (Reuss).
2. (Optional) Vitali's Reaction. — In an evaporating dish heat some of the dry alkaloid
with a few drops of fuming nitric acid to dryness. Moisten the yellow residue with
alcoholic KOHr reddish-violet color.
3. (Optional) Presence of Apoatropin in Atropin or Scopolamin. — To a solution of the
suspected alkaloid add a drop of i per cent, permanganate: the presence of apoatropin
causes an immediate reduction (brown precipitate). Pure atropin or scopolamin remain
clear (Kessel, 1906, Arch, intern. Pharmacod., 16, i).
4. (Optional) Distinction of Belladonna Bases. — This is made by the characters of
their gold salts.
5. (Optional) Isolation. — Proceed by Chapter IV, Exercise III, using sodium bi-
carbonate in No. 5.
6. (Optional) Solanin. — Preparation, Abderhalden's Handb., 2, 966; Tests, ibid.,
6, 133-
Physiologic Tests. — Dilation of pupils and paralysis of vagi.
TECHNICAL REFERENCES
Preparation of Atropin, Abderhalden's Handb., 2, 921; Quantitative recovery of atropin
from tissues, Fickewirth and Heffter, 1913, Bioch. Zs., 40, 37; Preparation of Scopolamin,
Abderhalden, 2, 927.
EXERCISE VH.— EPINEPHRIN, PHYSOSTIGMIN, PILOCARPIN, NICOTIN,
AMINS, AND RELATED ALKALOIDS
i. Epinephrin. — (a) Dilute solutions turn pink or brown on standing.
This is hastened by alkalies.
(b) To some i : 50,000 solution of epinephrin, or to a dilute extract of
suprarenal gland, add some ferric chloric!, drop by drop, as long as the color
darkens: a green color develops. Add some NaOH: the color changes to a
dark brownish red (Vulpian's Chromogen Reaction).
Physiologic Tests. — Vasoconstriction ; dilation of pupils; inhibition of
intestines or uterus.
TECHNICAL REFERENCES
Med
Bui. No. 100; Folin, Cannon and Denes, 1913, Jour. Biol. Chem., 13, 477; Vanderkleed,
1006, Jahrb. Pharm., 66, 263; Hale and Seidell (Krauss), Chem. Abstr., 7, 804, 1913,
U. S. P. IX.
2. (Optional) Physostigmin. — (Notice pinkish color.) (a) To i : 1000 aqueous solu-
tion add i drop of NaOH: red; becomes green on heating, and returns to red on cooling.
Add Sulphurous Acid: again colorless (Eber).
. (6) Evaporate some solution with a few drops of NH3: red color, leaving dry blue
residue. Add water: blue solution. Add Acetic Acid: violet in transmitted, coppered
fluorescent in reflected, light.
S. M.— Atropin.
S. M.— Epinephrin, i : 50,000.
CHAP. V SPECIAL TESTS OF IMPORTANT ALKALOIDS 5Q
•
Physiologic Test. — Constriction of pupil.
3. (Optional) Pilocarpin.— Shake some dry pilocarpin hydrochlorid with a granule
of potas. dichromate, 2 c.c. of chloroform, and i c.c. of 3 per cent. HjO2. The chloroform
acquires a blue or violet color. Apomorphin, strychnin, and antipyrin give rather similar
but distinct reactions.
4. (Optional) Nicotin. — (a) Equal volumes of ethereal solutions of nicotin and iodin
precipitate, changing gradually to large led needles (Roussin's crystals).
(b) Estimation in Tobacco. — Off. Agric. Chem.
Physiologic Tests. — Frog-position, tremors, vagus ganglia.
TECHNICAL REFERENCES
Preparation of Pilocarpin. — Abderhalden's Handb., 2, 963; of Hordcnin, ibid., 965;
Piperin, ibid., 917, Spartcin, ibid., 932; Coniin, ibid., 909; Arecolin, ibid., 014.
Estimation of Nicotin in Tobacco, etc. — Assoc. Off. Agr. Chem.; Abderhalden, 2, 916;
6, 128.
Isolation of coninm alkaloids from animal tissues, Billing, 1909, Bioch. Jour., 4, 286.
Cholin, Preparation and Tests. — Abderhalden's Handb., 2, 522; Renshaw, 1910, Jour.
Amer. Chem. Soc., 32, 128; Rosenheim, 1005, Jour. Ph., 33, 220; Kaufmann and Vorlaender,
1910, Zentr. Bioch. Bioph., n, 3; Physiologic Test, R. Hunt, 1915, Jour. Pharmacol.
Exp. Ther., 7, 307; Isolation, Stanek, 1006, Zs. physiol. Chem., 47, 83; 48, 334; Estimation,
Kinoshita, 1010, Arch. ges. Physiol., 132, 607; Ellinger, 1914, Muench. Med. Woch., 2336.
Betain.— Abderhalden's Handb., 2, 522; 7, 74.
Cytisin. — Preparation, Abderhalden, 2, 968.
Cod-liver Oil Bases. — Ibid., 2, 1042.
Amin Bases. — Preparation, ibid., 8, 261.
"Simpler Natural Bases." — Barger, 1914.
Vitamins. — Isolation, Sullivan and Voegtlin, 1916, Proc. Amer. Soc. Biol. Chem.,
3, 16-
EXERCISE Vin.— ACONITIN, VERATRIN, COLCfflCIN
1. Aconitin. — (a) Prickling Taste.-^-Note the taste of aconite (i : 300)
perceptible in dilution of i : 600 when 4 c.c. of the dilution is kept in the
anterior part of the mouth for one minute. This has been used for quanti-
tative estimation.
(b) (Optional). — There are no characteristic chemic tests for pure a« -unit in. but the
commrn ial Mimplrs generally give the following test for Pseudaionitin (Vital!
at<- the alkaloid with fuming nitric acid on water-bath, and moisten with alcoh« ii K» '1 1
red color, tinged with violet.
Physiologic Test.— Frog's heart.
2. Veratrin (Cevadin).— To a trace of powdered alkaloid add:
(0) A drop of concentrated H2SO4: yellow color. Apply lu-at : the color
changes through orange and deep scarlet to a beautiful violet red.
(b) A drop of concentrated HC1 and heat: red color (Trapp).
Physiologic Test. — Peculiar action on muse K .
Optional i Colchicin.— To a trace of the powdered alkaloid add a drop <>:
• sti-d Milphuri. a. ill: \vll..\\ -"lutinn. Add a trace of nit ri
.dkalin.- with Ko! I: yellowish red (Other . lu-mi. .md physiologic tests,
Foehner, 1010, Arch. exp. Path. Pharm., 63, y, \ carb. Amcr. Pharm.
Assoc., i, 417-
EXERCISE IX. -QUIPHN
Use a saturated aqueous solution of quinin sulphate.
i. Notice the l>lu< fluorescence, best seen by drawing the solution into a
pipet. This is increased by acids, diminished by NaCI.
5. J/.— Aconite, i : joo.
60 A LABORATORY GUIDE IN PHARMACOLOGY
Thalleioquin Reaction. — Add 2 drops of bromin water (enough to
give a permanent precipitate), and then cautiously an excess of ammonia.
An emerald color results, which is changed to red by HC1. (If a very small
quantity of ammonia is used, the color may be magenta.) (Brandes, Andre.)
3 (Optional) Herapathite Reaction.— To an alcoholic solution of quinin add some
iodin reagent (i part uxiin. i part 50 per cent. HI, 50 parts 70 per cent, alcohol, 0.8 part
sulphuric acid). Let stand: Crystalline plates, with green metallic luster, polarizing
* ^Optional) Quinin in Tablets.— Kxtract with a little water and apply Reaction 2.
(Optional) Determination in Urine.— See Nishi, 1909, Arch. Exp. Path. Pharm.,
60, 318; Baldoni, 191:. rcl., Xcntr. Bioch. Bioph., 14, 315; ibid., 17, 837; Abderhalden's
Handb., 3, 942.
TECHNICAL REFERENCES
Thalltioquin Reaction.— Fuehner, 1905, Arch. Pharm., 244, 602; technic, Abensaur,
Quantitative 'Estimation of Quinin.— Abderhalden's Handb., 6, 125; Dufilho, 1914,
Zentr. Bioch. Bioph., 16, 885.
Cinchonin in L'rinc. — Abderhalden, 3, 943.
Preparation of Cincltona Alkaloids.— Ibid., 2, 934.
Watson's Test for Cinchona Alkaloids— Add a few drops of alcoholic alpha-naphthol
containing 2 drops of concentrated sulphuric acid per i c.c. The cinchona alkaloids (and
no others) give a yellow precipitate, soluble in an excess of the reagent (Yearb. Amer.
Pharm. Assoc., 2, 418, 1913).
QUESTIONS ON CHAPTER V
(a) Describe a characteristic test for strychnin, morphin, and quinin.
(b) How would you isolate an alkaloid from a hypodermic tablet?
CHAPTER VI
SPECIAL TESTS FOR IMPORTANT GLUCOSIDS AND NEUTRAL
PRINCIPLES
EXERCISE I.— (OPTIONAL) DIGITALIS PRINCIPLES
1. Kiliani's Test. — Two solutions are used: (A) 100 c.c. concentrated sulphuric acid
with i c.c. of 5 per cent, ferric sulphate. (B) 100 c.c. glacial acetic acid with i c.c. of 5 per
cent, ferric sulphate. The digitaloid is dissolved in 3 to 4 c.c. of (B), and under this is
poured an equal volume of (A), and allowed to stand.
Digitoxin gives a dark contact zone and deep blue acetic layer.
Digitalin (true) colors the sulphuric acid yellow, red, and finally reddish violet.
Digitonin (pure) gives no color.
2. Keller's Test for Digitoxin. — Dissolve in glacial acetic acid containing a little
ferric chlorid. Float this on strong sulphuric acid: result as in i.
3. Strophanthin. — (a) K-strophanthin (official): Moisten the dry substance with
80 per cent, sulphuric acid: green color.
This test is also given directly by the seeds of Strophanthus Kombe" and hispidus.
However, it disappears with storage (Baldoni, 1915, Arch, di Farm., 19, 511).
(b) H-strophanthin: Moisten with concentrated sulphuric acid: red color.
(c) G-strophanthin (ouabain) : Dissolve in a little water and pour on C9ncentrated
sulphuric acid: acid pink to red; water dirty green.
4. Digitonin (and "German Digitalin") has the characters of saponin, lakes blood,
and forms a characteristic compound with cholesterin.
Physiologic Test for Digitaloids.— Frog's heart slowed, and systolic
standstill.
CHAP. VI IMPORTANT c.I.t COSIDS AND NEUTRAL PRINCIPLES 6 1
TECHNICAL REFERENCES
Chemic tests for Digitalis constituents, Kiliani, 1913, Amer. Jour. Pharm., 85, 223;
Dimethylamidobenzaldehyd as chemic test, Bufalini, 1913, Arch. Farmacogn., Sept. 15.
Isolation of Digitoxin from organs, etc., Gadamer, 433.
EXERCISE H.— (OPTIONAL) SANTONIN AND EMODIN CATHARTICS
i . Santonin Color Reactions. — (a) Dissolve a little in alcohol, add a small piece of dry
K< ill. and warm: reddish-green to yellow color (Banfi).
(b) To a trace of the dry substance add a little concentrated sulphuric acid and a
drop of ferric chlorid, and heat: dark red color, changing to violet brown.
(c) Rub a small quantity with KCN and heat: dark red mass, dissolving in water or
alkalies with irreen fluorescence.
I. Isolation from Tablets.— Kxt rait with chloroform, evaporate, and apply the tests.
3. Isolation from Feces or Gastric Contents. — Heat on water-bath with milk of lime
for several hours; strain; shake with benzol to remove impurities. Acidulate with IK 1
and extract with chloroform or benzol. Purify by crystallization from hot water or by
cautious sublimation between watch-glasses.
4. Santonin Urine. — None of the santonin appears unchanged in the
urine, but occurs there as Santogenin, probably as a combination of niono-
and dioxysantonin (Jaffe, 1897, Zs. physiol. Chem., 22, 538). The urine
polarizes to the left, and is yellow when acid, red when alkaline.
5. Distinction from Rhubarb and Similar Urines. — The urine after clirvs-
ophanic acid, rhubarb, senna, and other emodin cathartics is also yellow
when acid, red when alkaline. This may be distinguished from santonin
by the following tests:
(a) Sodium carbonate colors the rhubarb urine at once, santonin only
after a time. The red color is permanent with rhubarb, but disappears in
one or two days with santonin.
(b) Lime-water precipitates the red color with rhubarb, not with santonin
(Munk).
(c) Digestion with zinc dust decolorizes the red rhubarb urine, not the santonin.
(d) Ether shaken with the acid urine is colored yellow with rhubarb,
unchanged with santonin. On adding alkali to the decanted ether layer,
this turns red with rhubarb, but remains colorless with santonin (Pen-
zold;
(e) Amyl alcohol shaken with the alkaline urine takes up the color from santonin,
not from rhuharb ( Hoi»pe-Seyler).
6. Aloes in Urine.— The urine is shaken in a te-t -tul.r \\ith an equal vohfl
ether. The ether is decanted and evaporated, the residue dissolved in a little al-
and a trace of copper sulphate added: red color.
TECHNICAL REFERENCES TO SANTONIN
Test :rdt, 1007, U. S. I' I tigot, 404; Determination in Santonica, C. E.
Caspari, 1914, Jour. Amer. Pharm. Assoc., 3, 634.
Pdletierin. — Abdcrhalden's Handb., 2, 921.
EXERCISE in.— (OPTIONAL) PICROTOXIN
Vote the inteiiM-lv l-ilter l.i-tc (oMl «!n.|> of 1 tOOO tohltkx) • I
(6) Mix an equal quantity (Ira. ei <>l' pi. r««t..\in am! p. •••tassium i
t rated Milphuri. a. id. and then drop by drop, a Strong sodium hy-
drate -.. »lut ion- 1-ri. k red <
Physiologic Tett. IV. uliar i <>n\ ulsions of frog.
EXERCISE IV.— (OPTIONAL) CANTHARTOHf
Abderh.l !!>., 2, 889.
62 A LABORATORY GUIDE IN PHARMACOLOGY
CHAl'lT.k VII
SPECIAL TESTS OF IMPORTANT AROMATIC DERIVATIVES
EXERCISE I.— PHENOLS
The following tests are given in more or less modified form by all phenols,
although the typical colors apply only to phenol proper:
1 . Phenol.— Use a 5 per cent, solution.
(a) Add a trace of FeXl«: blue-violet color.
(b) Add bromin-water: yellow precipitate (tribromphenol) of needle-
shaped crystal (Landolt).
(c) Add Millon's reagent (mercurous nitrate) and heat: blood-red color
or precipitate (Plugge).
(</) (Optional) Azo-dye Reaction. — To a few cubic centimeters of i per cent, anilin
hydrochloric! add a few drops of concentrated HC1; cool on ice; add a few drops of 5 per
cent. NaNOj; make alkaline with 15 per cent. NaOH, and add the alkaline phenol solution:
brownish-yellow color. Acidulate with HC1: red precipitate.
(f) (Optional) Isolation of Phenol. — Acidulate with tartaric acid and distil with
steam.
(/) (Optional) Phenol Urines. — These are distinguished by their smoky color; richness
in ethereal and deficiency of inorganic sulphates; and sometimes contain free phenol.
(g) (Optional) Phenol Estimation in Urine. — Abderhalden, 3, 823; 5, 313; colorimetric,
Folin and Denis, 1915, Jour. Biol. Chem., 22, 305; Phenol and Paracresol, Siegfried and
Zimmermann, 1915, Bioch. Zs., 70, 124.
Methods of Phenol Estimation. — Forbing, 1916, Jour. Amer. Pharm. Assoc., 5, 166;
Permanganate method, Pence, 1913, Jour. Ind. Eng. Chem., 5, 218; Determination of
Phenol in cresol mixtures, Ditz and Bardach, 1912, Bioch. Zs., 37, 272.
(k) (Optional) Phenol Estimation in Tissues. — Sollmann, Hanzlik and Pilcher, 1910,
Jour. Pharmacol., i, 442; E. M. Mumford, 1913, ref., Yearb. Amer. Pharm. Assoc., 2, 382.
(i) Note that the reaction of strong carbolic acid to litmus paper is
neutral.
(j) (Optional) Phenyl-sulphonates. — Barium chlorid does not precipitate directly,
but (foes so after prolonged boiling with HC1.
2. (Optional) Cresols. — Creosote, Guaiacol, and Thymol give tests similar to those of
phenol.
(a) (Optional) Determination of Guaiacol in Urine. — Boil the urine with HC1; shake
with ether; evaporate ethereal layer, "dissolve in alcohol, and test with trace of ferric
chlorid: blue or green color.
(b) (Optional) Isolation of Thymol from Urine.— A. Seidell, 1915, U. S. Hyg. Lab.
Bui. 101, 43.
3. (Optional) Beta-naphthol. — This also gives similar reactions. The following are
distinctive: (a) It dissolves in alkalies with blue fluorescence.
(6) Dissolve in concentrated alkali, add a few drops of chloroform and heat: blue
color (Lustgarten).
(c) In urine: to 5 c.c. of urine add 3 or 4 drops of solution of chlorinated lime and a
few drops of concentrated HC1: lemon yellow color (naphthoquinon). Shake with
ether: this takes up the color. Pour this over i per cent, aqueous resorcin: red ring.
4. (Optional) Resorcin. — This gives the usual phenol tests, (a) The azo-dye reaction
is deep purple.
(b) It gives a pink color with NaOH and a trace of chloroform (Reuter).
(c) Isolation from urine: evaporate to one-quarter; boil with sulphuric acid; extract
with ether; evaporate ethereal layer. The resorcin is in the ethereal layer, and may be
purified with charcoal.
5. (Optional) Pyrogallol. — (a) Solutions are colored violet, brown, or black by lime-
water.
(/») It reduces solutions of silver and other metals.
(c) With formaldehyd and concentrated HC1 it gives a red color in the cold or on
gentle heating.
CHAP. VII IMPORTANT AROMATIC DERIVATIVES 63
6. Indol Reaction. — Baudisch, 1915, Zs. physiol. Chem., 94, 133; determination, Can-
telli, 1915, ref., Zentr. Bioch. Bioph., 18, 59.
EXERCISE U.— ANILIN DERIVATIVES
i. Common Tests. — The anilin derivatives, of which acetanilid and
acetphenetidin are the most important, give the indophenol reaction, which
depends on the amido group
(a) Indophenol Reaction. — Boil some acetanilid (or phenacetin) with a
little concentrated HC1 for one or two minutes (to liberate the anilin and
form paramidophenol). Cool; add an equal volume of 5 per cent, phenol
(to form indophenol) and a few drops of fresh solution of chlorinated lime:
red turbid fluid. Supersaturate with ammonia and shake: indigo blue
color.
2. (Optional) Distinctive Tests Between Acetanilid and Phenacetin.— (a) Heat some
acetanilid with XaOH solution: Dissolves, with odor of anilin; add a few drops CHClj and
heat again: Odor of phenyl-isonitril (resembles witch-hazel). This reaction is also given
by anilin, but not by phenacetin, etc. (Hofmann.)
(b) Rub together equal volumes of Acetanilid and NaNOi and add some concentrated
HjSO4: Orange liquid. Phenacetin gives a violet black color, later passing into green.
(c) Boil with HO and add a few drops of 3 per cent, chromic acid: acetanilid gives
a yellow color changing to green; phenacetin, ruby red.
3. (Optional) Isolation from Tablets, etc. — Kxtnu t with ether, evaporate and apply
the tests. Qitantitatii-c Estimation, Seidell, 1907, Jour. Amer. Chem. Soc., 29, 1091;
Kcblcr, Jour. Anu-r. I'harm. Assoc., 3, 1078, 1914.
4. (Optional) Isolation from Organs. — Proceed by Chapter IV, Exercise III, Nos. i
to 4. Watery extractions should be made hot.
5. (Optional) Tests in Urine. — These substances are excreted mainly
as paramidophencl, and therefore give the indophenol reaction: To about
10 c.c. of urine add \ volume of concentrated HC1; boil; allow to cool; add
J volume of 5 per cent, carbolic acid and a few drops of potassium bichro-
mate solution; red color: add ammonia: blue color.
6. (Optional) Anilin.— This gives the indophenol reaction without previous heating
with i in.
EXERCISE m.-ANTIPYRIN
1 . To an aqueous solution add a few drops concentrated FcjCl^: deep
red solution; + H2SO4: light yellow (Cohn, Knorr).
2. To an aqueous solution add sonu- Spirit us yEtheris Nitrosi. Slow
development of green color and precipitate of isonitroso-antijn riii.
3. (Optional) Antipyrin precipitates the alkaloidal pro ipitants.
4. (Optional) Test in Urine.- --Apply 'IV>t i dim tly (<> tin- uriiu-
5. (Optional) Determination in Tissues.— Lauber and \\intrr. 1913, ref., Chem.
Abstr., 7, 1729.
EXERCISE IV.— SALICYL DERIVATIVES
These give the reactions of phenols.
i. Sodium Salicylate.— (a) To a dilute solution add a drop of dilute
ferric chlorid: red violet color. (The reaction i- hindered !•
(b) 1 DM dry s;ili, \-late in test-tube; add equal pa ethyl
ai ! concentrated HaSO4 and heat: odor of methyl salicylate (oil of
\\intcrgreen).
5. M.— Chlorinated lime, fresh solution.
64 A LABORATORY GUIDE IN PHARMACOLOGY
(c) (Optional) Isolation from Tissues, Foods, <*<;.— Either by distillation of the acid
solution, or by extraction according to Chapter IV, Exercise III, Nos. i to 4.
Salicylic acid is often used as a food preservative (about 0.2 gm. per liter or kilo).
detected by the ferric chlorid test (a), but it must first be isolated in fairly pure form.
If the material is solid or semisolid, 200 to 300 gm. are hashed, triturated with 400 c.c. of
slightly alkaline water, ami >trained. This liquid (or the original sample, if it be liquid)
ii acidulated with sulphuric acid and extracted with ether or chloroform, and the ethereal
or chloroformic layer washed twii e with a little water. If the sample contained little or no
fat, this extract may be evaporated directly, at a low temperature, and the residue taken
up with hot water. This may be divided into several portions and used also for the tests
for beiuotc acid and saccharin. Since fruits may contain salicylic acid, not more than
50 c.c. of wine or £O gm. of fruit should be represented by the portion of the extract which
is used for the •flora te>t. If this quantity gives the test, one may be sure that salicylic
acid has been added. Only a or 3 drops of 0.5 per cent. Fe2Cl6 should be used.
Iftkf sample contains considerable fat, the ethereal or chloroformic solution is extracted
with dilute ammonia- water, the ammoniacal watery solution evaporated almost to dry-
ness, divided, and tested as ab<
For the detection in milk, 500 c.c. of the milk and 50 gm. of sand are evaporated
to diyness on a water-bath. The residue is extracted with acidulated alcohol. The
alcoholic nitrate is neutralized with ammonia, evaporated to dryness, dissolved, divided,
and tested as above.
( >ptionaJ) Demonstration of Salicyl in Esters (Wintergreen oil, salol, aspirin, etc.).
— Some give the iron reaction directly; all do so after saponification.
To a solution of the ester in water or dilute alcohol (or to the distillate of the organs
or extract) add a few drops of NaOH solution; boil a few minutes; add a drop of dilute
ferric chlorid; acidulate lightly with HC1; cool; neutralize carefully with ammonia:
violet color.
3. (Optional) Determination of Salicylates in Urine. — (a) Add a few
drops of ferric chlorid: violet color. This test is generally sufficient.
(b) Acidulate the urine and shake out with ether. Decant the solvent, and shake
it with very dilute ferric chlorid: violet color.
In place of the ether, a mixture of 3 parts of petroleum ether and 2 parts of chloro-
form may be used, which gives less emulsification; or a mixture of ether and benzol,
which dissolves also the salicyluric acid.
Salicyl urines polarize to the left, and reduce Fehling's feebly.
TECHNICAL REFERENCES ON SALICYLIC ACID
Isolation from Foods, Milk, Tissues, etc.—Offic. Agric. Chem.
Quantitative Determination in Organs, etc.— Bondi and Jacoby, 1905, Beitr. Chem.
raysiol., 7, 518; Seidell, 1909, Jour. Amer. Chem. Soc., 31, 1164; Cassal, 1910, Bioch.
< entr., 10, 674; Bondzynski and Humnicki, 1900, Jahrb. Pharm. 69, 218; Sauerland
Bioch. Zs., 40, 65, 1912.
Quantitative Estimation in Urine— Abderhalden's Handb., 3, 958; Gadamer «6-
bromin method, Lagrange, 1906, Paris Thesis.; Hanzlik, 1916.
Phenol Impurity.— Carletti, 1907, U. S. P. Digest, 127.
Salicyluric Acid Determination.— Baldoni, 1915, Arch, di Farm., 18, i.
EXERCISE V.— BENZOIC ACID AND SACCHARIN
i. (Optional) Benzoic Acid.— (a) To dilute solution of sodium benzoate add drop of
i' cfaJorid: brownish-pink precipitate. Add a little dilute HC1: dissolves.
:e precipitate of benzoic acid may be thrown down if the solution was concen-
trated.)
(b) Conrrrsion into Salicylic Acid.— To 10 c.c. of i : 1000 benzoic acid add 3 drops
a i : 10 dilution of Liq. Ferri Chloridi, then 3 drops of 3 per cent, ferrous sulphate,
king after each addition. A violet color develops in one-half to ten minutes; sensitive
0.1 to 0.2 mg. (Jonescu; Fleury; ref., Jahrb. Pharm., 73, 170, 1913).
. -Similar to salicylic arid. It may be separated from the latter by
bromin water, which does not precipitate benzoic acid.
I or tin- d<-tc< tion of benzoic acid used as zfood preservative (0.5 gm. per kg being the
usual quantity); this is isolated by the methods described under Salicylic Acid
employed * * C*Xntial that °nly ' °r 2 drops of a °'S Per cent ^^ Fe'C1«
CHAP. VII IMPORTANT AROMATIC DERIVATIVES 65
TECHNICAL REFERENCES
Determination in Foods. — Offic. Agric. Chem., Hilger, 1909, Jour. Ind. Eng. Chem.,
i, 538; Jonescu, Bioch. Centr., 8, 918; in catsup, La Wall and Bradshaw, Amer. Jour.
Pharm., 80, 171; in milk, Leach, Jour. Pharm., 1903, 486; in butter, Reinsch, Jour. Pharm.,
1003, 496.
Quantitative Estimation. — Folin and Flanders, 1911, Jour. Ame.r. Chem. Soc., 33, 161;
in urine, Raiziss and Dubin, 1915, Jour. Biol. Chem., 20, No. 2.
In L rine. — Abderhalden's Handb., 3, 831.
2. (Optional) Hippuric Acid. — Determination in urine, Abderhalden's Handb., 3,
828; 5, 315: rapid, 7. ;jo: Folin and Flanders, 1911, Jour. Amer. Chem. Soc., 33, 161.
3. (Optional) Saccharin. — This is sometimes used as an adulterant in sweets, etc.
It differs from sugars in being soluble even in ether and chloroform. If, therefore, the
substance is extracted with these solvents, and this solution evaporated at a low tem-
perature, a sweet taste of the residue indicates the presence of this adulterant. The
extraction may be made in a separating funnel, tjhe substance, if solid, being dissolved in
water. A i hemical test may be applied by heating this residue with NaOH to 210° C.,
whi<h eon verts it into salicylic and, characterized by the color reaction with iron. If
salicylic acid was present originally, this must first be destroyed by oxidation with KMnO4.
TECHNICAL REFERENCES
Abderhalden's Handb., 7, 356; Determination in foods, Offic. Agric. Chem.; Testoni,
Jahrb. Pharm., 69, 425; Estimation in urine and feces, Bloor, 1910, Jour. Biol. Chem., 8,
227; Wakeman, ibid., 8, 233.
EXERCISE VI.— (OPTIONAL) PICRIC ACID
i. Dye-test. — A woolen and a cotton yarn are left in the solution over night, and
washed. The wool is colored, the cotton not. This may be applied directly to the organ-
extracts, Ctc.
I. Isopurpuric Acid Reaction. — Heat the solution with KCN: blood-red color.
3. Isolation. — Chapter IV, Exercise III, Nos. i to 4.
4. Demonstration in Urine. — Boil the urine with HC1; extract with ether; evaporate
the ethereal layer and apply the tests.
EXERCISE VH.— (OPTIONAL) NITROBENZOL
Dissolve in alcohol; reduce to anilin with zinc dust and HC1 (one-half hour); make
alkaline with NaOH; extract with ether, and test for anilin.
EXERCISE Vm.— (OPTIONAL) ATOPHAN (PHENYL-QTJINOLIN CARBO-
XYLIC ACID)
Atophan urines give the following reactions (Skon/ew-ki and John, ion. \\icn.
klin. \Voi h., No. 49): (a) A few drops added to concentrated HC1 color this bright yellow.
(b) Pho>photiing>th ai id gives a yellow preiipitate.
ddition of ammonium sulphate with ammonia gives a dark green color.
The F.hrli. h diazo-reaction appears after twenty-four hours.
EXERCISE IX.— (OPTIONAL) PHENOLPHTHALEIN
\c id -olutinn-> arc « olorle^. but turn red with alkalies.
2. Tin- red ( olor disappears on heating \\ith /in. dust.
^timation in Tablets. — Kebler, Jour Amer. Pharm. Assoc., 3, 1096.
QUESTIONS ON CHAPTER VII
(a) Describe a characteristic test for phenol; acetanilid; antipyrin;
li( vlate.
(b) How would you isolate phenol from stomach contents?
(c) How would you isolate -alii ylic acid from urim •?
(d) How would you isolate acetanilid from a headache powder?
66 A LABORATORY GUIDE IN PHARMACOLOGY
CHAPTER VIII
SPECIAL TESTS OF IMPORTANT ALIPHATIC DERIVATIVES
EXERCISE I.— ETHYL ALCOHOL
Most of the tests are not distinctive, but are given by other alcohols,
aldehyds. esters, etc. Use (about) i per cent, solution for the following
tests:
i . Anstie Chromate Test.— Add some K2Cr2O7 solution and dilute H2SO4
and warm: green color and odor of aldehyd or acetic acid.
•kobol
coming more" intense, then 'fading. The reagent consists of 0.5 gm. potas. dichromate
gm. concentrated sulphuric acid. This is the most delicate test, sensitive to i : 10,000.
3. Lichen's lodofonn Test.— Add some NaOH and iodin solution; heat
gently: odor of iodoform; and precipitate of this substance may be seen
consisting of microscopic hexagonal plates. The test is sensitive to i : 5000
and is not given by pure methyl alcohol.
4. (Optional) Berthelot Test. — Add a little benzoyl chlorid, shake well, let stand a few
minutes, and add excess of KOH: odor of ethyl benzoate. Sensitive to i : 2000.
5. (Optional) Flame Test.— Place i pint of beer in a liter flask. Stopper tightly with
a perforated cork bearing an upright glass tube of a bore of £ inch and at least 4 feet high.
Heat slowly to boiling, and continue the heat until the foaming subsides. Apply a lighted
match to the upper end of the tube: The alcohol vapor will ignite, most of the watery vapor
being condensed in the long tube.
6. (Optional) Isolation from Tissues, etc., and Quantitative Estimation. — Make
strongly acid with phosphoric acid and distil until all the alcohol is removed (Test 2).
Filter the distillate (cotton in condensing tube). The alcohol percentage of the distillate
is calculated from its specific gravity (details, Hanzlik, i (c)).
A permanganate method for very small quantities is described by Barendrecht, 1913,
ref., Zentr. Bioch. Bioph., 14, 901. Estimation in blood, Abderhalden, 5, 195. Estimation
of small quantities of vapor, Baudrexel, 1911, Zentr. Bioch. Bioph., n, 543; Hamill, 1910,
Jour. Physiol., 39, 476; Abderhalden, 5, 1046; General Methods of quantitative determina-
tion, Abderhalden, 2, i.
7. (Optional) Estimation of Alcohol in Pharmaceutic Preparations. — See U. S. P. IX;
Vanderkleed, 1909, Amer. Jour. Phar., 81, 129.
8. (Optional) U. S. P. Purity Tests.
9. (Optional) Examination of alcoholic liquors, Abderhalden, 7, 339.
EXERCISE H.— (OPTIONAL) METHYL ALCOHOL
Distinction from ethyl alcohol is especially important.
1. Reduction Test. — Add i c.c. of i : 1000 potassium permanganate: methyl alcohol
is decolorized at once; ethyl only after twenty minutes.
2. Form aldehyd Test (Mulliken and ScuddeY). — Apply the following tests to two
solutions, one containing 10 per cent, of ethyl alcohol, the other 5 per cent, of methyl and
5 per cent, of ethyl alcohol. Determine which sample is adulterated. Place 10 c.c. of the
solution in a large test-tube. Heat a spiral of copper wire red hot and plunge into the
solution. Repeat this five or six times. (This converts methyl alcohol into f ormaldehyd ;
the further test is for this substance.) Filter. Boil very gently until the odor of acetalde-
hyd disappears. Pour into a test-tube and cool. Add i drop of 0.5 per cent, resorcin
solution; shake. Pour a portion of this liquid into a second test-tube containing concen-
trated sulphuric acid, held in an inclined position, so that the two liquids do not mix.
Let stand three minutes and rotate slowly: A rose-red ring indicates methyl alcohol (due
to formation of f ormaldehyd).
CHAP. VIII IMPORTANT ALIPHATIC DERIVATIVES 67
3. Formic Acid Test. — The methyl alcohol is oxidized by hydrogen peroxid into
formic acid, Schmiedel, 1913; ref., Yearb. Amer. Phar. Assoc., 2, 379.
4. Determination in Blood and Tissues. Nuloux, 1912 and 1913; ref., Chem. Abstr.,
6, 3102; and Zentr. Bioch. Bioph., 16, 158.
OTHER TECHNICAL REFERENCES
Bukowski, 1910, Centr. Bioch., 10, 55; Deniges, 1910, Zentr. Bioch. Bioph., 10, 300;
Simmonds, 1912, Amer. Jour. Phar., 85, 457; Szeberenyi, 1913, Zentr. Bioch. Bioph., 15,
635.
5. Tests of Methyl Alcohol in Liquors. — Vivario, 1914; ref., Zentr. Bioch. Bioph., 18,
620.
EXERCISE m.— (OPTIONAL) AMYL ALCOHOL (FUSEL OIL)
1. Marquardt Test. — Add a little water and i per cent, permanganate to red color.
Let stand for a day in stoppered vessel: valerianic odor.
2. Demonstration in Alcoholic Liquors. — HoLlaender, ref., Zentr. Bioch. Bioph,, 9,
783-
EXERCISE IV.— (OPTIONAL) ACETONE
i. Lichen's Test.— As for alcohol (Exercise I, No. 3). In distinction from alcohol,
acetone gives the test also with ammonia and ammonium iodid (Gunning).
I. Legal's Test. — Add a few drops of fresh sod. nitroprussid solution and make
alkaline with XaOH: red color (not given by alcohol); acidulate with acetic acid: carmin
color (difference from acetaldehyd, creatin, creatinin, and p-cresol).
3. Penzoldt Indigo Test. — Add saturated watery solution of o-nitrobenzaldehyd and
NaOH: yellow, then green color; after ten minutes, blue precipitate of indigotin, soluble
in chloroform. Not given by alcohol or acetaldehyd.
TECHNICAL REFERENCES
Acetone Substances (Acetone, Diacetic Acid) in Urine and Blood. — Abdcrhaldcn's
Handb., 3, 006, 921; 5, 197, 1222; Cervello and Girgenti, 1914, Arch. exp. Path. Pharm.,
>?; Marriott, 1913, Jour. Biol. Chem., 16, 281; in blood, 18, 508; Sammct. IQI
ppyaoL Chem., 83, 212; Folin, Jour. Biol. Chem., 3, 177; Folin and Denis, 1914, Jour.
Biol. Chem., 18, 263 (turbidity method).
Beta-oxybutyric Acid. — Abderhalden, 3, 924, 5, 199; Shaffer and Marriott
Jour. Biol. Chem., 16, 265; Marriott, ibid., 18, 508; Kcnnaway. 1014. Hiorh. Jour., 8, 230;
Folin and Denis, 1914, Jour. Biol. Chem., 18, 263; Shaffer and Hubbard, 1916, Proc.
Amer. Soc. Biol. Chem., 3, 27; Van Slyke, 1916, Proc. Soc. Exp. Biol. Med., 13, 134.
EXERCISE V.— (OPTIONAL) ETHER
The chemic tests are not characteristic. The purity tests of the U. S. P. may be
applied. (An extensive discussion of these is given by Baskerville and Hamor, 1911,
Jour. Ind. KIIK. Chem., 3. 301, 378.)
Estimation of Ether. — Nicloux, 1006, Bioch. Centr., 6, 48. Determination in .iir.
Koc hmann am! '912, Zentr. Bioch. Bioph., 14. 14.
EXERCISE VI.— CHLOROFORM
i. Schwarz's Reaction. — To ;i -nlution of chloroform add a
c of resorcin and a few drops of NaOH, and heat: pink color.
>ptional) Lustgarten's Reaction.— Dissolve o.i jrm. of alpha-naphthol in
beat to 50 < ;m<lad<l afewdropsof thcsuspo t«l solution: blue color. A. i.:
•cd j>rc< i|>:
.v (Optional) Hoffmann's Reaction.— Heat gently with alcoholic NaOH and a few
: rilr o<lor.
4. (Optional) Isolation. Distillation of tlic a. i.lul.itrd material.
Opttoaal Quantitative Estimation.— Decomposition of vapors by combustion
with Ca<>. <«r boiling \\jth a). ..h..|i, K< »H.
6. (Optional) Purity Tests of U. S. P.
68 A LABORATORY GUIDE IN PHARMACOLOGY
TECHNICAL REFERENCES
Estimation in'Air and Vapors.— Kix hmunn and Strecker, 1912, Zentr. Bioch. Biop>,.
14, 14; Hewitt Anesthetics, 26; Mavelung, 1910, Arch. cxp. Path. Pharm., 62, 414; Nicloux,
1910, Zbl. Bioch. Bioph., 10, 495.
Estimation in Blood.— Loth, ion, /»>!. Hioch. Bioph., 12, 239.
Waller Gas Balance.— Hewitt Anesthetics, 103; Boothby and Sandiford, 1914, Jour.
Pharmacol., 5, 369.
Alcohol in Chloroform— Nicloux, 1005, Jahrb. Pharm., 66, 170.
Distinction Chloroform and Chloral.— Jona, 1911, Chem. Abstr., 6, 1337.
EXERCISE VII.— (OPTIONAL) CHLORAL HYDRATE
This gives all the reactions of chloroform.
1. To a water>- solution add NaOH: odor of chloroform.
2. Nessler's Reagent gives a brick red precipitate, gradually changing to yellowish
green (difference from chloroform).
3. Isolation. — Distillation of acidulated material.
4. Chloral Urine. — The chloral is excreted mainly as urochloralic acid (trichlorethyl
glycuronic acid), which is decomposed by boiling with dilute acids into trichlorethyl alco-
hol and glycuronic acid. The urine, therefore, gives the Fehling test and polarizes to the
left. Urochloralic acid is isolated by the method of Kuelz (Arch. ges. Physiol., 33, 221)
or Mchring and Musculu3 (Gadamer, 295; Abderhalden's Handb., 3, 970).
EXERCISE Vm.— (OPTIONAL) SULPHONAL
The dry powder is decomposed by heating with:
1. Powdered wood charcoal: formation of a mercaptan (odor) and formic acid (litmus).
2. Reduced iron: mercaptan odor; residue with HC1 yields HjS.
3. KCN: mercaptan odor and KSCN (extract gives red color with ferric chlorid).
4. Isolation.— Proceed by Chapter IV, Exercixe III, Nos. i to 4. Watery extracts
must be filtered hot.
5. Determination in Urine. — Morro, 1894, Deut. Med. Woch., 34.
EXERCISE IX.— (OPTIONAL) VERONAL
1. Acidulate a saturated solution with HC1 and add a few drops of Millon's reagent:
white gelatinous precipitate, soluble in excess of the reagent.
2. Isolation from Tissues or Urine. — Proceed by Chapter IV, Exercise III, Nos. i to 4.
TECHNICAL REFERENCES
Isolation and Detection. — Gadamer, 458; Panzer, 1908, Bioch. Centr., 8, 167; Heidu-
schka, Jahrb. Pharm., 71, 463; Macadie, Chem. Abstr., 7, 1526.
EXERCISE X.— (OPTIONAL) ALDEHYD REACTIONS
1. Nessler's reagent gives a yellowish-red color, gradually changing to black, espe-
cially on heating.
2. Ammoniacal silver solution is reduced in the dark (silver mirror).
3. Fuchsin-sulphurous acid is gradually colored red.
EXERCISE XI.— (OPTIONAL) PARALDEHYD
1 . This gives all the aldehyd reactions.
2. It gives the Lieben and Legal tests; see Acetone, Exercise IV.
EXERCISE XH.— FORMALDEHYD
This gives all the general aldehyd reactions. The special reactions may
be divided into those which occur with weakly alkaline reaction (i, 2, and
3); strongly alkaline reaction (4), and strongly acid reaction (5 and 6).
Since the stronger reagents may liberate formaldehyd from its compounds,
only the first class (Nos. i to 3) can be used when testing for free formalde-
hyd in the presence of its derivatives (hexamethylenamin, etc.).
In the following tests use i : 50,000 solution of formaldehyd (i drop of
official liquor per liter).
S. If .—Formaldehyd, i : 50,000; Jorissen phloroglucin reagent; phenylhydrazin hydrochlorid,
0.5 per cent;, sod. nitroprussid, 5 per cent.
CHAP. VIH IMPORTANT ALIPHATIC DERIVATIVES 69
1. Jorissen Phloroglucin Test. — To i or 2 c.c. of the suspected solution
add 0.5 c.c. of the reagent (phloroglucin o.i gm. in 10 c.c. of 10 per cent.
NaOH; keeps well) : pink to red color, becoming more intense, then gradually
fading. The test is sensitive to i : 10,000,000, and may be applied directly
to all body fluids, even when tinged with blood, but not to bile or undiluted
blood.1 The test may be simplified by adding a trace of dry phloroglucin
to the fluid after making this distinctly alkaline with NaOH.
2. Rimini Phenylhydrazin Test (Burnam's Test). — To about 10 c.c. of
the suspected fluid add 3 drops of 0.5 per cent, phenylhydrazin hydrochlorid;
2 drops of 5 per cent. sod. nitroprussid ; and 3 drops of 10 per cent. NaOH:
emerald green to deep blue color, changing to orange or red. Water alone
gives a greenish-yellow color with the test, changing more rapidly to red.
Formaldehyd urine may first give a purple color. The test is sensitive to
i : 1,000,000 and may be applied directly to all body fluids except bile and
whole blood (Hanzlik).
3. (Optional) Phenylhydrazin-ferricyanid Test.— Substitute 5 per cent, ferricyanid
for the. nitroprussid in the Rimini test: red color. More delicate.
4. (Optional) Lebbin's Test. — To about 10 c.c. of the suspected fluid add 0.5 gm. of
resorcin and an equal volume of 50 per cent. NaOH; boil: red color.
5. Liebermann's Test. — Mix some of the formalin solution with a drop
of 5 per cent, phenol and pour cautiously, without mixing, on some con-
centrated H2SO4 in test-tube: crimson zone.
6. HehneVs Test.— To about 5 c.c. of the solution add i c.c. of milk or
peptone solution. Pour this on an inch of concentrated sulphuric acid con-
taining a trace of ferric chlorid : violet zone; this test may be applied directly
to suspected milk, by pouring this on the ferric-sulphuric acid.
7. (Optional) Formation of Hexamethylenamin. — This occurs when formaldehyd
solution is evaporated with ammonia. It may be recognized by the precipitation reac-
tions.
8. (Optional) Isolation of Formaldehyd. — Distillation of weakly acid liquid; 300 c.c.
of the liquid material (or if solid, 200 gm. moistened with 100 c.c. of water) are acidulated
with phosphoric acid and distilled, collecting the first 40 to 50 c.c. This is then tested
either by Hehner's method or by any of the other tests.
9. (Optional) Quantitative Estimation. — Collins and Hanzlik, 1916, Jour. Pharm-
acol., 8, 130.
TECHNICAL REFERENCES
Tests.— Abderhalden's Handb., 2, 14; Dunning, 1913, Amer. Jour. Pharm., 85, 453:
Hald, 1911, Arch. exp. Path. Pharm., 64, 329.
EXERCISE Xm.— HEXAMETHYLENAMIN (UROTROPIN)
Use (about) i : 100 solution.
1. Bromin Precipitation.— To the solution add bromin- water, drop by
drop: orange precipitate, which mli-solves until more of the reagent is
added. This and the other precipitation tests are not given by
maldehyd. They may be applied to normal urine, but not to any fluids
containing proteins.
2. (Optional) Alkaloidal Precipitants. Prr. ipitates arc given with men-urn- fhlorid,
Millet hosphomolybdii, and other alkaloidal precipitants.
5. If.— Milk; formaldehyd milk (o.i c.c. formald. sol. per liter).
' Hanzlik and Collins, Arch. Int. Med.. 1913, /*, S7&
yO A LABORATORY GUIDE IN PHARMACOLOGY
3. Liberation of Formaldehyd.— Hexamethylenamin is decomposed by
acids into formaldehyd and ammonium. It therefore gives the formaldehyd
5 and 6 directly, but i and 2 only after treatment with acids.
(a) Render the hexamethylenamin solution freely acid with HC1 and
boil (or let stand in stoppered test-tube) : odor of formaldehyd. To some
of this solution add excess of NaOH: odor of ammonia. Use the remainder
of the solution for (6).
(b) Apply the Jorissen test ( i) to some of the boiled acid solution : positive.
(c) Apply the Jorissen test (i) and Liebermann's test (5) to some of the
fresh hexam. solution: i is negative, 5 is positive.
4. (Optional) Tests in Urine.— Hexamethylenamin is excreted as such
by the kidneys, and gives the bromin test (i) directly. In acid urines,
a small quantity of formaldehyd is liberated continuously, giving the
Jorissen and Rimini tests (Exercise XII, Nos. i and 2). Alkaline urines
do not give this test, but respond to Liebermann or Hehner tests (Exercise
XII, Nos. 5 and 6).
Acid hexamethylenamin urine does not usually show bacterial turbidity
when kept for a day in the incubator.
5. (Optional) Quantitative Methods.— Falk and Sugiura, 1916, Jour. Pharm. Exp.
6.' (Optional) Test for Hexam. in Blood or Bile. — Acidulate, distil, and test for for-
maldehyd.
TECHNICAL REFERENCES
Determination in Galenic Mixtures. — Puckner and Hilpert, 1908, Jour. Amer. Chem.
Soc., 30, 1471.
EXERCISE XIV.— (OPTIONAL) FORMIC AND ACETIC ACIDS
These are the only volatile aliphatic acids of toxicologic importance.
They are distinguished by their characteristic odor and taste.
1. Ferric chlorid, in neutral solution, gives a red color with both. On heating, the
solution darkens and then gives a brown precipitate.
2. Mercuric chlorid, on boiling, is reduced to calomel (white precipitate) by formic
acid, not by acetic.
3. Mercurous nitrate, on warming, is reduced to metallic mercury by formic acid.
Acetic acid does not reduce, but on cooling concentrated solutions deposit crystalline
plates of mercurous acetate, soluble on heating.
4. Silver nitrate is also reduced by formic acid, not by acetic.
5. Dry sodium acetate, heated in a test-tube with equal volumes of alcohol and con-
centrated sulphuric acid, gives the odor of ethyl acetate (acetic ether). Formate gives a
different odor (rum) and evolution of CO.
6. Quantitative Test for Formates in Food. — Croner and Seligman, 1907, Bioch.
Centr., 6, 306.
7. Quantitative Estimation of Formic Acid in Urine, etc. — (Pohl) Sollmann, 1908,
Jour. Amer. Med. Assoc., 51, 821; Franzen and Greve, 1909, ref., Amer. Pharm. Assoc.,
58, 355; Freyer, 1895, Chem. Ztg., No. 51, 1184; Dakin, Janney and Wakeman, 1913,
Jour. Biol. Chem., 14, 341.
TECHNICAL REFERENCES
Acetic Acid. — Abderhalden's Handb., 2, 20.
EXERCISE XV.— (OPTIONAL) VALERATES
Dilute sulphuric acid liberates valeric acid, of characteristic odor.
EXERCISE XVI.— (OPTIONAL) CITRATES, TARTRATES, OXALATES, AND
OTHER ORGANIC ACIDS
i. Citrates. — Calcium chlorid does not precipitate in the cold, but gives a white
granular precipitate on boiling. Isolation from other acids: Albahary, Zentr. Bioch.
Bioph., 13, 337; Tests, etc., Abderhalden's Handb., 2, 32.
CHAP. VIII SPECIAL TESTS OF IMPORTANT ALIPHATIC DERIVATIVES 71
2. Tartrates. — These give a white crystalline precipitate with potassium salts. Am-
moniacal silver nitrate solution gives a metallic silver mirror on heating. Further tests,
etc., Abderhalden's Handb., 2, 32.
3. Oxalates. — (a) To a solution of potassium oxalate add Cad: Precipitate. Add
acetic acid: does not dissolve. Add dilute HC1: solution.
(b) Isolation of Free Oxalic Acid. — Mix the material with sand and dry on water-bath,
pulverize, and extract with boiling alcohol for several hours (reflux condenser). Filter;
make slightly alkaline with KOH and boil for one-half hour. Dilute with water and
evaporate the alcohol. Acidulate with acetic acid and precipitate with CaCU (let stand).
\Ya>h the precipitate with hot water; boil with sodium carbonate; filter; neutralize with
acetic acid and precipitate the oxalate with lead. Filter; suspend the precipitate in
water; and decompose with H2S. Filter and crystallize.
(c) Isolation of Soluble Oxalates. — The material left from the alcoholic extraction in
(2) is extracted with water, boiled, and the protein precipitated with acetic acid. The
filtrate i.- precipitated with CaClj, etc., as in (2). Details, Gadamer, 400; Tests, etc.,
Abderhalden's Handb., 2, 40.
4. Lactic Acid. — Abderhalden, 2, 28; determination, Wolff, 1914, Jour. Physiol.,
48, 341; in organic material, Bellet, 1913, Zentr. Bioch. Bioph., 15, 556, 635; in tissues and
fluids, Yoshikawa, 1913, ibid., 16, 10; Meissner, 1915, Bioch. Zs., 68, 175; in urine, Ryffel,
1709, ibid., 10, 384; in Wood, Abderhalden, 5, 194; infeces, ibid., 5, 387.
5. Malic Acid.— Abderhalden, 2, 34.
6. Succinates. — Ibid., 2, 24.
EXERCISE XVIL— (OPTIONAL) FATTY ACIDS AND FATS
1. Volatile Fatty Acids.— Abderhalden's Handb., 5, 386.
2. Butyric Acid. — Ibid., 2, 20.
3. Oleic Acid. — Determination, Polano, Zs. Geburtsh. Gyn., 65, 584.
4. Fats.— Abderhalden's Handb., 2, 199; 7, 184; Determination, Kumagawa-Suto
method, ibid., 5, 476; infeces, ibid., 5, 363; Saxon, 1914, Jour. Biol. Chem., 17, 09; Laws
and Bloor, 1916, Amer. Jour. Dis. Child., n, No. 3; rhanges by freezing, Smith, Miller
and Hawk, 1915, Jour. Biol. Chem., 21, 395; in blood, Abderhalden's Handb., 5, 161;
Bloor, 1914, Jour. Biol. Chem., 17, 377; in milk, Bloor, 1914, Jour. Amer. Chem. Soc., 36,
1300; iodin and s a ponifi cation values, U. S. P. 1 V
5. Glycerin. — Isolation and tests, Gadamer, 388; Reactions, Deniges, IQOQ, Jahrb.
Pharm., 69, 173; Ganassini, 1913, Zentr. Bioch. Bioph., 14, 772; in blood, Abderhalden,
5, 196; Determination in galenicals, Briggs, 1915, Jour. Amer. I'harm. Assoc., 4, 75; Bradts,
ibid., 4, 78.
6. Acrolein.— Qualitative, Ganassini,. 1913, Zentr. Bioch. Bioph., 14, 772.
EXERCISE XVIIL— (OPTIONAL) LIPOLDS
"Lipoids" are the intracellular substances soluble in fat solvents, but exclusive of
simple fats and fatty a. id>. They consist chiefly of lecithin and rholest. : pins"
cover all substances soluble in fat soh
Preparation.— Abderhalden's Handb., 5, 613; Nerve, ibid., 2, 774; Brain, Mathews,
I'hy-iol. Chem., 875, 15.
Phosphatids.— Ibid., 2, 256; Solubility, il.id.. 5. 548; Part:
Separation «.f l.ipin- from l.ipin Extracts, EtOMDMOOm, 1914. Soc. Exp. Biol. Mcd
98; Ccrcbrositli, Smith and Mair, mi i. Zentr. Biiwh. Bioph.. n, 540.
CholMtwin.— AbderhaJden'i Handb., 2, 244; QiMflltetiM Estimation, Wask.
i'.ith. Phar:
> hn-il.rr, iliid .. 15, 788; Thaison and i
panson ; \\Vltmann. 1913, WtOL Klin Wocfa., \; .ilc • olorimet ru ); in blood,
Abderhalden's Handl... ;. 166; Hl.x.r, 1916, Jour. Biol. Chem.. .'4. N rytkrocytes,
ibid., 5. 20?; in f>< • t. ibid.. ;. }66.
Lecithin. \\ I h, 1006, Zs. physiol. Chem., 4:
and \Vo,,,i HIT, Iti-.l. Chen . i. \ -.-.Lawsonai '
zow, I9M, An h. int. I'harmacod., lood-OOfpUKkl. Abdefhald
f..r hy|HMl«-rmir use, Mondnhi. Chem. Al^tr. 1911, 6. i ; ?7: l^trrmination and emul
tion. s, hi /s., 40, 189; in blood, Abderhalden, 5, 166; Bloor, 1915,
Jour. Hi--] <
Phytosterin. Demonstration in .mim.d Eats, Kuchn, Bcngcn, and Wcwcrinkc, 1915,
ref.. "I'''-, 18, 362.
7 2 A LABORATORY GUIDE IN PHARMACOLOGY
EXERCISE XIX.— HYDROCYANIC ACID
1. Notice odor (which, however, may be confused with benzaldehyd or
nitrobenzol).
2. Schonbein Reaction.— Impregnate some filter paper with freshly
prepared Tincture Guaiac, let dry, then pour on some very dilute CuSO4;
expose this to the vapor of i : 1000 HCN: deep blue color (Pagenstecher,
Schonbein, I 'never). Expose another paper prepared in a similar manner
to the vapor of*NHs: green color.
This test can be applied directly to suspected material, stomach washings,
etc. A negative reaction definitely excludes HCN; but a positive reaction
is not distinctive: the reaction depends upon the liberation of ozone by the
interaction of HCN and CuSO4; and ozone may be formed in other ways.
3. Berlin-blue Reaction. — Add to i : 1000 solution of HCN some FeSO4
and Fe^Cla and a few drops of NaOH; boil, let stand a few minutes, acidulate
with concentrated HC1, and heat: green to blue color, or precipitate of ferric
ferrocyanid (Husemann, Ittner).
4. (Optional) Liebig Sulphocyanid Reaction. — Render the solution slightly alkaline
with NaOH, add a little yellow ammonium sulphid, and evaporate on water-bath. Dis-
solve in water, acidulate with HC1 and add a drop of dilute ferric chlorid: red color of
ferric sulphocyanid.
5. (Optional) Isolation of HCN. — The material is acidulated with tartaric acid and
distilled. The HCN is in the first fractions of the distillate.
The presence of sulpho-, ferro-, or ferricyanids could give rise to errors, since these
may be partly decomposed in the distillation. If their presence is demonstrated (color
reactions with ferric chlorid), the liquid is made alkaline, heated to 60° C. and the HCN
carried over with a current of CO2 (Jacquemin-Otto) .
6. (Optional) Determination of Small Quantities. — Viehover and Johns, 1915, Amer.
Jour. Phar., 87, 261; in plant tissues, Alsberg and Black, 1916, Jour. Biol. Chem., 25,
No. i.
7. (Optional) Estimation in Organs. — Waller, 1910, Jour. Physiol., 40, xlvii.
8. (Optional) Sulphocyanids. — These give a red color with ferric salts after acidula-
tion with hydrochloric acid. Tests and quantitative estimation, Abderhalden's Handb.,
3, 259; in saliva, Autenrieth and Funk, 1912, Muench. med. Woch., 59, 2657, 2736; Gies
and Kahn, 1913, Chem. Abstr., 7, 1049.
EXERCISE XX.— (OPTIONAL) CARBON DISTTLPHID
1. Heat a few drops with alcoholic lead acetate: black color of PbS.
2. Evaporate a few drops with alcoholic ammonia on water-bath to dryness. Forma-
tion of sulphocyanid, which gives red color with ferric chlorid.
EXERCISE XXI.— (OPTIONAL) PIPERAZIN (DIETHYLENDIAMIN)
1. Reactions. — Precipitation by alkaloidal precipitants; especially characteristic is
a scarlet red crystalline precipitate with bismuth-potassium iodid.
2. Demonstration in Urine. — Add a little NaOH to precipitate earthy phosphates.
Filter; render filtrate weakly acid with HC1, warm to 40° C. and add bismuth potassium
iodid solution. If amorphous precipitate occurs at once, filter. The characteristic
crystalline scarlet red precipitate appears after a time.
QUESTIONS ON CHAPTER VIII
1. How would you test a solution for the presence of alcohol?
2. How would you test stomach contents for the presence of chloroform
or chloral?
3. How would you test milk for formaldehyd?
4. How would you test hexamethylenamin urine — (a) for hexamethyl-
enamin; (b) for free formaldehyd; (c) for bound formaldehyd?
5. How would you test stomach contents for cyanid?
S. M.—Tr. guaiac; HCN, i : 1000.
CBAP. IX SPECIFIC TESTS OF IMPORTANT HEAVY METALS 73
CHAPTER IX
SPECIFIC TESTS OF IMPORTANT HEAVY METALS
The ordinary tests for inorganic substances are so well covered in the
usual courses of qualitative analysis that they need not be repeated. Those
which are of especial medical interest are cited, mainly for convenient refer-
ence. Their special application to the urine is practically important. The
substances are arranged alphabetically in each chapter.
All the exercises of this chapter are optional.
EXERCISE I.— ALUMINUM
1. Reactions.- XaOH gives a while precipitate, soluble in excess; ammonia, a white
>itate insoluble in excess.
2. Alum in Baking Powders.— Incinerate about 2 gm. Extract with boiling water
and filter. Add to filtrate a few drops of ammonium chlorid solution: flocculcnt pre-
cipitate indicates alum (Off. Agric. Chem.).
». Isolation. — Destroy organic matter by Fresenius-Babo. Precipitate with am-
monia. Dissolve in NaOH; reprecipitate with ammonium chlorid.
Determination in Fcces. — Schmidt and Hoagland, 1912; Jour. Biol. Chem., n, 387.
Estimation in Tissues. — Gies et alias, 1916, Bioch. Bui., 5, 151.
EXERCISE H.— ANTIMONY AS TARTAR EMETIC
1. Mineral acids precipitate antimonous acid (SbO»Ha), soluble in excess.
2. Alkalies precipitate the oxid, SbiOj, soluble in excess of KOH or NaOH, not in
carbonates or ammonia.
3. Hydrogen sulphid gives a yellow color in neutral solutions, an orange precipitate
in the presence of HC1.
4. Estimation. — Cloetta, 1911, Arch. exp. Path. Pharm., 64, 352; Brunner, 1912, Ibid.,
68, 186.
EXERCISE m.— ARSENIC
i. Reduction Test for Solid Arsenic Trioxid. IM.ue ixnvder in the botton of diffi-
cultly fusible test-tube shown in Fig. 4. In the «>nst ruled portion place a splinter of
freshly roasted wood charcoal. Heat the charcoal to redness, then the arsenic: this is
>=o
Fig. 4.— Arsenic reduction tube.
volatilized and reduced to As in passiim over the carbon, and condenses in the cold parts
of the tube to a black mirror. In the upper part- it i- oxidized to an* I and
deposited as a white octa&lral sublimate. There is also the ihar. odor.
-senic Solutions.— (a) Hydn»««-n -ulplr'd ^ives lemon-yellow color or preti;
dissolving (olnrlcss in ammonium carbonate.
(6) Aiidulate with nitri. a. id; add silver nitrate; filter if necessary; and J*
filtrate dilute ammonia \\ it limit mixing lemon yellow /..me of silver arsenite. soluble in
ammonia and nit ri<
Reinsch's Test.- This may be applied also to impure solution lip of
thin bright . «.pper foil • about i .'m. square) in a test-tube with 10 rated
If the reagents are pure \dd some of the suspected liquid and
i irk stain may denote As, Sb, Sn, Hg, Bi; no stain proves
>f these n •
: from other metal- the foil i- pla.nl in a narrow test-tube and
heated: tl -l.iiili/es .<. ler parts as As or A-
4. Biologic Test. Culture- of iVni. illium bre\ i. aule. \\hen growing on arsenic
a garlic «!• >t is chara« \ imulated onl> by tellurium
74 A LABORATORY GUIDE IN PHARMACOLOGY
and selenium. Under proper conditions it is extremely sensitive (to o.ooi nig.) and ap-
plicable to impure solutions. Technic, Abderhalden's Handb., 5, 3.
5. Marsh Test.— See page 52. The material must be free from organic matter.
Details, Gadamer, 155. The evolution of hydrogen is facilitated by first laying the zinc
in a solution of CoCU, acidulated with sulphuric acid.
6. Isolation of Arsenic from Tissues, Urine, etc.— See page 52.
;. Quantitative Estimation.— Gadamer, 168; in organs and tissues, Joachimoglu,
1014. Arch. exp. Path. Pliarm., 78, i.
8. Arsenic in Wall Paper, etc.— A piece of the paper is ignited and the flame ex-
tinguished: the plowing paper has the garlic odor. Positive result shows dangerous
quantity of arsenic. Smaller amounts may be demonstrated as in the tissues.
9. Arsenic in Pharmaceutic Preparations, etc. — See U. S. P. IX.
EXERCISE IV.— ORGANIC ARSENIC DERIVATIVES
1 . Cacodylic Acid.— (a) Reactions.— This does not give the arsenic tests, except after
decomposition by the Kjeldahl process. Solutions treated with zinc and sulphuric acid
(or phosphorous acid in the case of urines) give the characteristic odor of cacodyl oxid.
(6) Isolation from Urine (Vitali). — Urine of patient receiving cacodylate. Render
acid and concentrate; add equal volume of chloroform and sufficient alcohol so that the
fluids mix. Then add enough water to separate the chloroform. Draw off and evaporate
the chloroform. Test residue for cacodylic acid as in i (a).
(e) Estimation in Urine. — References, Merck's Report, 1910, 24, 6.
2. Atoxyl. — This gives the Reinsch and Marsh tests for As directly, and the tests for
aromatic amido groups. The solution, when mixed with a few drops of sodium nitrite
and HC1, forms a diazo compound which gives the following reactions:
(a) With alkaline phenol solution to alkaline reaction: purple color.
(6) With alpha-naphthylamin hydrochlorid: purple color.
(c) With beta-naphythylamin hydrochlorid: brick red color.
(6) and (c) are made more certain by the presence of urea.
(d) Detection in Urine.— Urine of patient receiving atoxyl. Apply the diazo tests as
above. If the urine is deep colored it may be partly decolorized with a little boneblack.
(e) Estimation. — Engelhardt and Winters, 1915, Jour. Amer. Phar. Assoc., 4, 1468.
3. Salvarsan. — (a) Abelin Test for Urine. — Urines of patients receiving salvarsan
or neosalvarsan give the following test, although it is doubtful whether this is specific:
Acidulate about 8 c.c. of the urine with 5 or 6 drops of dilute HC1 and add 3 or 4 drops of
0.5 per cent, sodium nitrite. Add a few drops of this mixture to 6 c.c. of a colorless alkaline
solution of resorcin: red color in the presence of salvarsan (Muench. med. Woch., 1911,
1002 and 1566).
(b) Determination in Tissues. — Richter, 1911, ref., Chem. Abstr., 5, 2396.
EXERCISE V.— BISMUTH
1. The insoluble bismuth salts, when dissolved in just sufficient warm nitric acid
and poured into a large excess of water, give a white precipitate, insoluble in tartaric acid.
2. Hydrogen sulphid gives a black precipitate.
EXERCISE VI.— CHROMIUM
1. Chromates give a yellow precipitate with barium or lead salts; a red precipitate
with silver or mercurous.
2. To about 5 c.c. of hydrogen peroxid solution in a test-tube add a little sulphuric
acid, a thin layer of ether, and a trace of the chromate: blue color of the ether (hyper-
chromic acid).
3. Isolation. — Destruction of organic matter, see page 52. Evaporation to dryness;
fusion with saltpeter; solution in water.
EXERCISE VH.— COPPER
1. Reactions. — Ammonia gives a deep blue color; ferrocyanid a red brown precipitate;
metallic iron acquires a coating of metallic copper.
2. Test in Stomach Contents. — Acidulate with HC1 and place in platinum crucible
with a piece of metallic zinc: copper deposit on platinum.
3. Isolation from Tissues, etc. — Destruction of organic matter, see page 52. Pre-
cipitation with hydrogen sulphid; solution in nitric acid; evaporation to dryness; solution
•m water.
CHAP. IX SPECIFIC TESTS OF IMPORTANT HEAVY METALS 75
EXERCISE Vni.— IRON
The medicinal iron preparations are either salts of iron, or the iron is a firmly bound
constituent of the molecule. The first class (inorganic irons) give the ordinary iron
reaction; the latter (organic or masked iron) do not.
i. Reactions of Ferrous Salts. — Hydrogen sulphid gives a greenish-black precipitate
of FeS, soluble in mineral acids; alkalies a white precipitate turning blue, green, and
brown; potassium ferrocyanid a white precipitate; ferricyanid a blue precipitate; sulpho-
cyanid no color.
j. Reactions of Ferric Salts. — Hydrogen sulphid as for ferrous. Tannin gives a blue
or greenish-black color; alkalies a brown pnvipitate; ferrocyanid a blue precipitate; ferri-
cyanid a brown color; sulphocyanid a blood- red color, bleached by mercuric chlorid, not
by alcohol. The red ferric sulphocyanid is extracted by ether.
3. Reactions of Salts with Organic Acids. — "Scale-salts," such as ferric citrate or
ferric ammonium citrate or tartrate, do not precipitate with hydrogen sulphid, ammonia,
or ferro- or ferricyanid. They are precipitated by NaOH (ferric hydroxid). After acidu-
lation, they are also precipitated by ferrocyanid.
4. Distinction of Ionic (Inorganic) and Non-ionic (Organic or Masked) Iron. — (a)
ilium's Reaction. — This is the most delicate: a drop of fresh i percent, hematoxylin
solution gives a blue-black color with inorganic iron, but not with organic. The test is
best applied to the dry substance or concentrated solution. Confirm that the following
preparations are correctly classed:
Inorganic: Ferric sulphate. Organic: Dried blood.
Scale salt of iron. Egg-yolk.
Iron albuminate. somatose.
(ft) The action of dilute hydrochloric acid liberates the inorganic iron from some of
the masked compounds, but not from others. To demonstrate this, add a little 5 per cent,
hydrochloric acid and a drop of potassium ferrocyanid to ovoferrin and to egg-yolk, and
boil: the first gives the Prussian blue reaction, the second not.
Lay some alcohol hardened sections of spleen in the ferrocyanid, and others in the
acid-ferrocyanid mixture. Spleen contains loosely bound organic iron (ferratin) and there-
fore colors in the acid mixture, but not in the plain ferrocyanid.
TECHNICAL REFERENCES
Abderhalden's Handb., 5, 1101; Estimation of traces, Jahn, 1911, Zs. physiol. Chem.,
78, 308; in presence of organic matter, Salkowski, Ibid., 43, 142.
EXERCISE IX.— LEAD
i. Lead Acetate. — This gives a yellow precipitate with dichromates or iodids; a white
pro ipitate with sodium hydroxid, chlorids, or bromids; a black prei ipit.ite with hydrogen
Milphid. The subacetate is also precipitated by acids.
»-ad Carbonate (White Lead).— (a) Heated with sodium carbonate by blow-
pipe, it fuses to the diutile metal and a yellow dcjM.Mt. Tin- metal may be dissolved
in nitric at id and te-ted a- lead at elate.
(6) It i> blackened l>y hydrogen Milphid.
3. Isolation from Foods, Tissues, Feces, Urine, etc.— Dcstrm lion of ori-ani.
see page 52. Solution «,f the PI»C13 in hot water; neut r.ili/at i««n by ammonia to weakly
ai id reaction; pro ipitation with hydrogen sulphid; solution in nitri<
TECHNICAL REFERENCES
[3, Bio.h. />.. ;;o. ;;o; I-'riedmann. I..IL /.- phv-iol. Chem.. QJ, 46;
< Him State Board of Ind 1th Hazards, pp. 187 and 388.
4. Detection in Drinking-water. Hl.i. k pn •• ipit.itr \\ilh hydrogen Milphid shows
Her <|uantities may I i by adding sodium phos-
tn 10 liters of the
the pre. ipit.iie in dilute nitric acid, cva|x>rating the excess of add. and precipitating with
hydrogen Milpl
xcess of Lead in Vessels, Solder, etc. H«>il for on, h.df hour with 4 per
te the soluti- n: pre. ipii.iie with hydr«>gci
6. Colorimetric Estimation of Traces.— Siegfried and Pozzt, 1914, Bioch. Zs., (••
76 A LABORATORY GUIDE IN PHARMACOLOGY
EXERCISE X.— MANGANESE
i . Reactions of Permanganates. — The color of the aqueous solution is discharged by
oxidizable substance In alkaline reaction there is also a brown precipitate.
KOH changes the color of permanganate to green, with evolution of oxygen.
olation of Manganese.— Destruction of organic matter, see page 73- Precipita-
tion with ammonium sulphid (flesh colored in pure solutions); fusion with sodium car-
bonate and nitrate: green fusion mass if manganese is present.
EXERCISE XI.— MERCURY
i . Reduction by Copper ( Applicable to Impure Solutions).— Reinsch test, see page 50:
gray deposit. Dry and rub lightly with filter paper: silver color. Heat in narrow test-
tube: stain disappears and is deposited on tube as gray mercury mirror; magnification
shows Hg globules. Place tube in stoppered flask containing a little iodin. In a few
hours red mercuric iodid is formed (or the original copper foil with its deposit may be
laid on a glass slide, next to a small piece of iodin, and covered with a watch-glass).
Reduction by Tin. — To a solution of mercuric chlorid add a fresh solution of
stannous chlorid: white precipitate of HgCl. More of the reagent, with heat, gives a
gray precipitate of Hg.
3. Klein's Test. — To the mercurial solution add a little KI, a drop of ammonium
chlorid, and then NaOH, drop by drop: brown or yellow color or precipitate (NHg2I).
This test is very delicate and may be made still more so as a contact method, adding the
NaOH containing NH4C1 without mixing.
4. Isolation of Mercury from Tissues or Urine. — Destruction of organic matter, see
page 50. Precipitation by H2S. Solution hi HC1 with KC1O3; evaporation at 50° to
60° C. Solution of the residue in water is suitable for the preceding tests.
Mercuric chlorid, iodid, or cyanid may be extracted directly from the dried material
by ether.
5. Determination of Minute Traces. — Strzyzowski, ref., Chem. Abstr., 7, 805.
6. Quantitative Estimation in Urine or Tissues. — As sulphid or electrolytic.
TECHNICAL REFERENCES
Buchtala, 1913, Zs. Physiol. Chem., 83, 212 and 249. Gadamer, 213; In urine,
Robert, Intox., 2, 335; Abelin, Zentr. Bioch. Bioph., 13, 829; Siebert, Ibid., 10, 434;
Klotz, 1914, Zs. Physiol. Chem., 92, 286; Perelstein and Abelin, 1915, Muench. med.
Woch., Aug. 31 (highly delicate test by precipitation with basic lead acetate); destruction
of organic matter, E. Salkowski, Bioch. Zs., 61, 27.
7. Quantitative Estimation in Bichlorid Tablets. — Chapin, 1914, Amer. Jour. Pharm.,
86, i ; La Wall, 1914, Jour. Amer. Phar. Assoc., 3, 50; Kebler, Ibid., 3, 1087, 1091.
8. Calomel. — (a) Lime-water gives a black mixture.
(6) KI solution gives a yellow, green, gray, or black color.
(c) Estimation: Grantham, 1915, Jour. Amer. Pharm. Assoc., 4, 441; in Tablets,
Kebler, 1914, Jour. Amer. Pharm. Assoc., 3, 1089.
EXERCISE XH.— PHOSPHORUS
1. Scherer's Preliminary Test. — Place some phosphorus water in a small bottle;
stopper it loosely and between the cork and the neck of the bottle suspend two pieces of
filter paper, the one impregnated with Silver Nitrate, the other with Lead Acetate If
the silver paper is blackened and the lead paper not, the presence of Phosphorus is rendered
probable. (If both are blackened, this indicates H^S.)
2. Luminous Ring Test. — See page 50.
3. Fresenius-Neubauer. — The material, in a flask, is acidulated with sulphuric acid
and distilled at 60° to 70° C. in a current of CO2. The vapors are passed through 3 per
cent, silver nitrate: phosphorus causes a precipitate.
A hydrogen apparatus is arranged as in the Marsh test and the hydrogen is ignited.
The silver precipitate is introduced into the flask. The flame is colored green if phos-
phorus is present.
4. Determination of White Phosphorus: Engelhardt and Winters, 1915, Jour. Amer.
Pharm. Assoc., 4, 451; in Matches: Phelps, 1914, Hyg. Lab. Bui. No. 96.
EXERCISE XHI.— SILVER
i. Reactions of Silver Nitrate. — NaCl gives a white curdy precipitate, insoluble hi
nitric acid, soluble in ammonia. This solution, heated with formaldehyd, deposits a
metallic mirror.
CHAP. X SPECIAL REACTIONS OF EARTHY AND ALKALI METALS 77
2. Isolation from Tissues, etc. — Destruction of organic matter, see page 52. The
precipitated AgCl is collected. Any remaining in solution is precipitated by hydrogen
sulphid, dissolved in nitric acid, evaporated to dryness, and precipitated with HC1. The
united AgCl is dissolved in ammonia and tested with hydogen sulphid, aldehyds, and
blowpipe fusion with KC'N i silver granule).
3. Determination of Traces.- MalaU--tu, 1015. nf., Zentr. Bioch. Bioph., 18, 85.
4. Determination of Ag in Protein Compounds. — Incineration (2 gm.); solution of
residue in warm, dilute nitrii arid. Titration with sulphocyanid.
5. Determination in Colloid Silver Preparations. — Dankwortt, 1915, ref., Zentr.
Bioch. Bioph., 18, 252.
EXERCISE XTV.— ZINC
1. Reactions of Soluble Zinc Salts. — (a) White precipitate with ammonium sulphid,
insoluble in acetic acid, soluble in HC1. (b) White precipitate with ferrocyanid, soluble
in KOH.
2. Zinc Oxid. — Turns lemon yellow on heating. Easily soluble in dilute acid, giving
the reactions of soluble zinc salts. Also soluble in NaOH.
3. Isolation. — Destruction of organic matter, see page 52. Addition of excess of
sodium acetate; precipitation (hot) with hydrogen sulphid; solution in nitric acid; conver-
sion into oxid by incineration; solution in dilute acetic acid.
EXERCISE XV.— ANALYSIS OF RARE ELEMENTS
Abderhalden, 8, 269.
EXERCISE XVI.— TESTS FOR HEAVY METALS
(See U. S. P. DC for "Limit test.")
QUESTIONS ON CHAPTER IX
1. How would you test a powder suspected of being arsenic trioxid?
2. Give an outline of the Marsh test.
3. Give an outline of the Reinsch test.
4. How would you test stomach contents for copper?
5. Describe a test for ferric salts.
6. Describe a test for ferrous salts.
7. Describe a test for differentiating organic iron.
8. How would you test a tablet for the presence of mercuric chlorid?
9. How would you determine whether a white powder is calomel?
10. How would you recognize phosphorus in stomach contents?
CHAPTER X
SPECIAL REACTIONS OF EARTHY AND ALKALI METALS
The cations are arnmged alphabetically. All the exercises of this
chapter are optional.
EXERCISE I.-AMMONIUM
.mi mi .iiium salts evolve ammonia vapors having the character-
istic odor and bluing litmus.
TECHNICAL REFERENCES
•ttimi. \l><!rrh. iMrn'~ Il.iiHll)., 3, 765; in wriV. Il'id . <;. :*=;: r.ipi*'
blood, Ibid., 5. 15*.; rapid. 7. 7-\ R..M-nb1.»..m. iQU, clinical for ;
Jour. Amer. Med. Assoc., 61, 87; substitute for NcssUr's Reagent, S. S. Graves, i
Amer. Chem. Soc.
78 A LABORATORY GUIDE IN PHARMACOLOGY
EXERCISE H.— BA1UUM
1. Reactions. — (a) White precipitate with sulphates (even calcium sulphate solution),
insoluble in dilute acid. (6) Bichromate gives a yellow precipitate, insoluble in acetic
acid (difference from lead), (c) The nitrate and chlorid color the Bunsen flame green.
2. Differences from Strontium. — The latter is not precipitated at once by calcium
sulphate. It i> not precipitated by dichromate. It colors the flame red.
3. Isolation of Barium.— Destruction of organic matter, see page 52; the insoluble
residue is saved. In the filtrate the greater part of the acid is neutralized, and the Ba
precipitated with sulphuric acid. The precipitate is added to the original insoluble residue;
dried and incinerated; oxidized with nitric acid; again heated to redness, and fused with
potas. sodium carbonate. The mass is extracted and the precipitated barium carbonate
dissolved in dilute HC1.
EXERCISE III.— CALCIUM
Ammonium oxalate gives a white precipitate, insoluble in acetic acid, soluble in HC1.
White precipitates are also given by sodium carbonate or phosphate; both precipitates are
soluble in acid. The sulphate precipitate is insoluble in acids.
TECHNICAL REFERENCES
Estimation in Urines, Organic Fluids, etc.: Abderhalden's Handb., 5, 293; Gut-
mann, 1914, Zentr. Bioch. Bioph., 16, 359; Goy, 1913, Ibid., 16, 359; Bell, 1912 (clinical),
Bioch. Jour., 6, 205; Stransky, 1914, Arch. exp. Path. Pharm., 78, 122; v. d. Heide, 1914,
Bioch. Zs., 65, 363; H. Lyman, 1915 (rapid method), Jour. Biol. Chem., 21, 551; in blood,
Halverson and Bergeim, 1916, Proc. Am. Soc. Biol. Chem., 3, 22; microcolorimetric,
Rowland, Haessler, and Marriott, 1916, ibid., 3, 18.
EXERCISE IV.— LITHIUM
1. Reactions. — Precipitate on warming with sodium phosphate, but not with car-
bonate or sulphate. It colors the flame crimson, with characteristic spectrum.
2. Detection in Urine. — Evaporate and incinerate. Extract with dilute HC1; evapor-
ate; extract with alcohol; evaporate: spectrum test. Quantitative Estimation, Murmann,
1910; Chem. Abstr., 5, 2607.
EXERCISE V.— MAGNESIUM
Sodium phosphate with ammonium chlorid and ammonia give a white crystalline
precipitate. NaOH or carbonate cause precipitation. No precipitate is given by bicar-
bonate, sulphate, or oxalate. ' Estimation in Urine, Abderhalden's Handb., 5, 293; in
tissues, Stransky, 1914, Arch. exp. Path. Pharm., 78, 122.
EXERCISE VI.— POTASSIUM
1. Reactions. — Lilac tint to colorless flame. Tartaric acid gives white crystalline
precipitate. Platinic chlorid gives yellow crystalline precipitate of potassioplatinic chlorid.
Sodiocobaltic nitrite solution gives yellow precipitate.
2. Quantitative Determination in Urine. — Abderhalden's Handb., 5, 292; 5, 1113;
H. J. Hamburger, 1915 (traces), Bioch. Zs., 71, 415.
EXERCISE VII.— SODIUM
Yellow tinge to flame.
EXERCISE Vm.— STRONTIUM
Crimson flame. Sulphates give white precipitate, soluble in strong acids. Distinc-
tion from barium, see Exercise II. Isolation, as for barium.
QUESTIONS ON CHAPTER X
1. How would you determine whether a cough mixture contains an
ammonium salt?
2. How would you determine whether a cathartic salt is magnesium
sulphate, sodium sulphate, or sodium phosphate?
CHAPS. XI, XII SPECIAL REACTIONS OF INORGANIC ACID RADICALS 79
CHAPTER XI
CAUSTIC MINERAL ACEDS AND ALKALIES; PEROXTOS
Toronto
All the exercises are optional.
EXERCISE I.— FREE MINERAL ACIDS
These need only be considered if the reaction is strongly acid to litmus. In the case
of organs an aqueous or alcoholir i-xtrac t is used.
i . Demonstration of Free Mineral Acid. — (a) M ethyl-mokt Test. — A i per cent, methyl-
violet solution is diluted with water to a linht violet color. Mineral acids change the color
to blue, green, and yellow. With oxalic acid the yellow is indistinct.
(b) lodin Test. — A very dilute solution of ferric acetate, mixed with KI and starch
solution, is gradually colored blue when notable quantities of free mineral acid are present.
Distinction of the Acid Radicle. — This may be done as in Chapter XII.
3. Demonstration of Free HC1. — This is important, since Cl ions are present in all
tissues. In the absence of other volatile miiu-ral adds this may be done by heating the
sample on the water-bath with a drop of methyl-violet: the green color would gradually
return to violet if the acidity was due to HC1.
EXERCISE H.— CAUSTIC ALKALIES
These need only be considered if the reaction is freely alkaline to litmus. Their
quantity is determined volumetrically in an aqueous extract, directly, and after preceding
precipitation with barium chlorid (to determine the share of the carbonates). The
cathion is identified as in Chapter X.
EXERCISE m.— HYDROGEN PEROXID
i. It evolves oxygen on contact with a crystal of permanganate.
Dilute solutions do not liberate iodin from KI and starch, but do so on adding a
crystal of ferrous sulphate.
3. The solution is rendered acid with sulphuric acid, a drop of very dilute dichromate
is added, and the mixture shaken at once with ether: the latter is colored blue.
QUESTIONS ON CHAPTER XI
How would you determine whether the strongly acid reaction of a
bloody vomitus is due to mineral acid?
CHAP I I K XII
SPECIAL REACTIONS OF INORGANIC ACID RADICALS
The anions are arranged alphabetically. All the exercises are optional.
EXERCISE I.— BICARBONATES
TheM ! the .arlx.Matt^. I. ul <!•• not |>ro ipit.itr earthy metals from
•'til li.-ilnl 'Ix.ilm- convert! tlu-m into carbonates).
EXERCISE II.— BORATES AND BORIC ACID
i. Reactions. (./) l'l,imr 7VW. Over a little l»..ri. idd 'ith a
<lr<'|» of -iil|iliu! .in evaporating «li»h p !• »li"l .u»<l iirnr
(6) / I'rst.— Turn <-d in bori lx>rate
;u i'lulated with HC1) and dried, turns brownish red. Ammonia changes this to bluish
80 A LABORATORY GUIDE IN PHARMACOLOGY
2. Determination in Urine, Tissues, Food, etc. — (a) Rough Method. — Add -jV of con-
centrated HC1 to the suspected fluid (e. g., milk with 1.5 gm. per liter) or extract, and
apply the turmeric test i (6).
(b) Exiifl Method. — About 25 gm. of the material (more in the case of urine) is made
alkaline with time-water, evaporated to dryness, incinerated, extracted with 15 c.c. of
ml sufficient HC'l to make it freely acid. Apply turmeric test i (ft).
3. Quantitative Estimation.— Off. Agr. Chem.; F. C. Cook, 1916, Jour. Agr. Res.,
5. 877.
EXERCISE in.— BROMIDS
1. Reactions.— (a) Silver nitrate gives a yellowish-white precipitate (AgBr), insoluble
in dilute nitric acid, soluble in ammonia.
(6) Chlorin-water in solutions acidulated with sulphuric acid liberates bromin, which
dissolves in chloroform with yellow color.
2. Detection in Urine. — Evaporate and incinerate at low heat. Extract with water
and test by i (6). If large quantities are present, this test may be applied directly to the
urine.
3. Quantitative Estimation in Urine, Blood, and Tissues. — Bernouilli, 1913, Arch.
Exp. Path. Pharm., 73, 365; Larsson, 1913, Bioch. Zs., 49, 479; Bogdandy, 1913, Zbl.
Bioch. Bioph., 15, 59; Autenrieth and Funk, 1912, Muench, med. Woch., 59, 2657, 2736
(colorimetric) ; Takeda, 1911, Arch, internat. Pharmacodyn., 21, 203; Bermann, 1910,
Ther. Mon., 183 (urine); Wyss, 1906 and 1908, Arch. exp. Path. Pharm., 45, 266; 49, 186;
Halogens in lipoids, Cappenberg, 1912, Chem. Abstr., 6, 1014. In medicines, Leech,
1915, Rep. Chem. Lab., A. M. A., 8, 54.
4. Isolation of Free Bromin from Stomach Contents, etc. — Passage of current of air
through material in a flask, catching the bromin in water, and shaking this with chloro-
form.
EXERCISE IV.— CARBONATES
1. White precipitates, soluble in dilute nitric acid, are given with the salts of Ca, Ba,
Mg, Pb., etc.
2. Acids liberate CO2 gas, which precipitates lime-water, but not calcium chlorid.
EXERCISE V.— CARBONIC ACID
1. Reactions. — See Exercise IV, 2.
2. Excess of CO2 in Air. — Large excess is demonstrated by the extinction of a candle
flame. Quantitative estimation by Pettenkofer's barium method (Gadamer, 49).
TECHNICAL REFERENCES
Estimation, Abderhalden's Handb., 3, 600; in blood, Ibid., 5, 157; minute quantities,
Tashiro, 1913, Amer. Jour. Physiol., 32, 107, 137; alveolar air, Y. Henderson and Russell,
1912, Ibid., 29, 436; Comparison of methods of obtaining, Boothby and Peabody, 1914,
Arch. Int. Med., 13, 497. Indicator method, Haas, 1916, Sci.. 44, 105.
Carbon. — Graphic demonstration in lung, E. F. Hirsch, 1916, Jour. Amer. Med.
Assoc., 66, 950.
Gas Analysis. — Abderhalden's Handb., 3, 555; 5, 1027 (Zunz); Haldane, 1898, Jour.
Phsyiol., 22, 465; Abderhalden, 3, 683; Table of Volume Reduction, Abderhalden, 3, 590;
micro-analysis, Ibid., 3, 658.
Preparation of Gases. — Ibid., i, 215, 230; Air analysis, Heinz, 2, 452.
Blood gases. — Abderhalden's Handb., 3, 664; Heinz, 2, 437; Tigerstedt, 2.1, i; Brodie,
1910, Jour. Physiol., 39, 391; Brodie and Cullis, 1908 (saline), Ibid., 36, 405; Absorption
and Tension, Fahr, 1910, Jour. Physiol., 43, Abderhalden, 3, 699; in circulating blood,
Ibid., 3, 703.
Intestinal Gases. — Abderhalden, 5, 415.
Oxygen Estimation. — Ibid., 3, 622. Determination of oxygen content of water, Wink-
ler-Hyman, Amer. Jour. Physiol., 40, 241, 1916.
EXERCISE VI.— CHLORATES
i. Reactions.— (a) Evolution of Cl gas on heating dry chlorate with concentrated
HC1.
(6) Added to sulphuric acid and indigo solution, there is no change; but decoloriza-
tion occurs when a trace of sulphurous acid is added.
CHAP. XII SPECIAL REACTIONS OF INORGANIC ACID RADICALS 8l
(c) Chlorates do not precipitate silver nitrate, but do so on the addition of sodium
bisulphite.
2. Isolation of Potassium Chlorate from Stomach Contents or Tissues. — Extract by
dialysis; concentrate by evaporation; precipitate with alcohol; crystallize from water.
3. Detection in Urine. — Test i (6) may be applied directly to the urine (Rabuteau).
4. Isolation from Urine. — Decolorize with lead subacetate. Remove lead from
filtrate with hydrogen sulphid; evaporate and crystallize.
5. Quantitative Estimation in Urine. — Hildebrandt, 1006, ref., Biocb. Centr., 5, 831.
EXERCISE VH.— CHLORIDS
Silver nitrate gives a white curdy precipitate, insoluble in nitric acid, soluble in
ammonia.
Lead salts also give a white precipitate, soluble in boiling water, insoluble in am-
monia.
TECHNICAL REFERENCES
Michrochemid Abderhalden's Handb., 5, 1131; Estimation in Urine, Ibid., 5, 291;
Symes, Jour. Physiol., 32, 221; simplified, McLean and Selling, 1914, Jour. Amer. Med.
Assoc., 62, 1081; in presence of SCN, Cormimboeuf, 1912, Chem. Abstr., 7, 39. In Blood,
etc., Abderhalden, 5, 207; Gazzetti, 1913, Zbl. Bioch. Bioph., 15, 791; McLean and Van
Slyke, 1915, Jour. Biol. Chem., 21, 223, 361, 509. Cl ions in blood, Abderhalden, 7, 727.
Centrifugalion method, Sueyoshi, 1916, ref., Jour. Amer. Med. Assoc., 66, 929.
EXERCISE Vm.— CHROMATES
(See page 74.)
EXERCISE IX.— FLUORTOS
1. Reactions.— (a) Dry NaFl, moistened with concentrated sulphuric acid, evolves
HI1. which etches glass.
(6) Solutions give a white precipitate with Ca or Ba salts.
2. Determination in Blood. — Abderhalden, 5, 159.
3. Detection in Foods.— Detection of I-luorids (used as food preservatives) : 150 c.c.
of the sample (or in the case of solid foods, the aqueous extract) are brought to boiling,
and mixed with 5 c.c. of 10 per cent. K2SO4 and 10 c.c. of 10 per cent, barium acetate.
The precipitate is allowed to settle, collected on a small filter, washed, and im im-rated in
a platinum crucible. A glass plate is coated with wax and some marks scratched through
the wax with a pointed stick. The contents of the crucible are moistened with com cn-
t rated HSSO4, and covered with the waxed plate, the edges of the crucible being firmly
embedded in the wax. The glass is now covered with a cooling device and the * nu il>!r is
heated for an hour as high as is possible without melting the wax. The gla— plate is now
< d and cleaned (by steam): a di>tin. t ct< hing proves that fluorin was present
EXERCISE X.— GLYCEROPHOSPHATES
1. They do not give a precipitate with ammonium molybdatc in the cold, but do so on
heating.
he dry salts, when strongly heated, evolve inflammable vapors and leave a
residue of pyrophospliatr.
3. A saturated aqueous solution of calcium glyceropbcMiphate deposits white irri-
descent scales of anhydmu- < alcium glyceropl, hoiling.
EXERCISE XI.-HYPOCHLORITES
These evolve Cl gas (odor) on the addition ••(
EXERCISE XH.-HYPOPHOSPHTTES
:th Milphuri. a. id and nii\nl with silver nitrate give a white
prn ipit.itr. . h.n (fag rapidly to blown <>r »>U k by rnlin ti
2. Solut i t sulphate produce a red-brown precipitate of cuprous
3. Ca or Ba salts are not precipitated.
82 A LABORATORY GUIDE IN PHARMACOLOGY
EXERCISE Xm.— IODIDS
1. Reactions. — (u) Silver nitrate gives a yellow precipitate, insoluble in dilute nitric
acid, practically insoluble in ammonia.
(6) Lead acetate give* a yellow precipitate, soluble on heating.
(c) Mercuric chlorid gives a red precipitate, soluble in excess of either reagent.
(d) To an iodid solution add a little sodium nitrite and dilute sulphuric acid: liberation
of iodin, with yellow or brown color, dissolving in chloroform or carbon disulphid with
violet color, and turning starch solution blue.
(c) Chlorin-water also liberates iodin.
2. Qualitative Test in Urine or Saliva. — To about 5 c.c. of the urine or
saliva add a few drops of concentrated sulphuric acid and of i per cent,
sodium nitrite solution. Shake out with chloroform: violet color of the
chloroform.
3. Quantitative Estimation of lodids in Tissues, etc. — Hanzlik, 1910, Jour. Biol.
Chem., 7, 259; in blood, Abderhalden's Handb., 5, 160; in lipoids, Cappenberg, 1912, Chem.
Abstr., 6, 1014. Electrolytic determination, Krauss, 1916, Jour. Biol. Chem., 24, No. 3.
4. Estimation of Organic Iodin in Thyroid. — Hunter, 1910, Jour. Biol. Chem., 7, 321;
Kendall, 1911, Proc. Soc. Exp. Biol. and Med., 8, 120; 1915, Jour. Biol. Chem., 19, 251;
Bernier and Perron, 1911, Zentr. Bioch. Bioph., 12, 57; F. C. Koch, 1913, Jour. Biol.
Chem., 14, 106.
5. Estimation in Presence of Bromids. — Bray and MacKay, 1910, Jour. Amer. Chem.
Soc., 32, 1193; Kendall, 1912, Chem. Abstr., 6, 2867.
EXERCISE XIV.— IODIN
1. Reactions. — (a) Odor, color, and violet vapor on heating.
(6) Blue color with starch solution, discharged by thiosulphate.
(c) Dissolves in chloroform or carbon disulphid with violet color.
(d) Forms iodoform with NaOH and alcohol.
2. Detection in Stomach Contents. — (a) Brown color of protein material, discharged
by thiosulphate or ammonia.
(6) Violet color of chloroform extract.
3. Stains. — As in 2 (a).
EXERCISE XV.— NITRATES
1. Reactions. — (a) To a 5 per cent, solution of potassium nitrate add an equal volume
of concentrated sulphuric acid; cool, and drop in a crystal of ferrous sulphate: dark brown
color of ferric sulphate around the crystal.
(b) Mix a nitrate solution with solution of ferrous sulphate and add a layer of con-
centrated sulphuric acid: brown ring.
(c) Add a drop of diphenylamin solution and a layer of concentrated sulphuric acid:
deep blue contact ring.
(d) Free nitric acid (or nitrate with HC1) discharges the color of indigo on heating.
2. Detection of Nitrate in Stomach Contents or Urine. — Evaporate the alkaline watery
extract or urine to dry ness; dissolve in a little water and test by i (a).
3. Quantitative Estimation in Urine. — Caron, ref., Chem. Abstr., 6, 2442.
4. Potassium Nitrate in Meat. — Off. Agr. Chem.
EXERCISE XVI.— NITRIC ACID
1. Reactions.— See Nitrates, Exercise XV.
2. Stains. — Yellow color of organic tissues, deepened to orange by ammonia.
Isolation from Stomach Contents. — Extract rapidly with alcohol; neutralize with
calcium carbonate, filter, and evaporate the alcohol.
EXERCISE XVH.— NITRITES
Acid solutions liberate iodin from KI and decolorize permanganate.
Estimation of Nitrous Orid. — Abderhalden's Handb., 3, 655.
Analysis of Nitrous Oxid. — Boothby and Sandiford, 1915, Amer. Jour. Physiol., 37,
371-
Nitroglycerin. — Estimation of small quantities, Scoville, 1911, Amer. Jour. Pharm.,
83, 3595 in tablets, Kebler, 1914, Jour. Amer. Pharm. Assoc., 3, 1094.
CHAP. XII SPECIAL REACTIONS OF INORGANIC AGED RADICALS 83
EXERCISE XVHI.— PERMANGANATES
(See page 76.)
EXERCISE XIX.— PHOSPHATES (ORTHO-)
1. Magnesia mixture gives a white crystalline precipitate.
2. Silver nitrate gives a yellow precipitate, soluble in ammonia and nitric acid.
3. Ammonium molybdate and heat gives a yellow precipitate.
Estimation in Urine. — Abderhalden's Handb., 5, 290; total phosphorus in tissues,
etc., A. E. Taylor and Miller, 1914, Jour. Biol. Chem., 18, 215; Neumann, 1902, Zs. physiol.
Chem., 37, 129; Chapin and Powick, 1915, Jour. Biol. Chem., 20, No. 2; Forbes, Beegle,
and Wussow, Ohio Agr. Exp. Sta. Tech. Bui. 8. Microcolorimetric method for serum,
Rowland, Haessler, and Marriott, Proc. Amer. Soc. Biol. Chem., 3, 18, 1916.
EXERCISE XX.— SILICATE (SODIUM)
Acids produce a gelatinous precipitate of silicic acid.
Determination in Urine. — Salkowski, Zs. physiol. Chem., 43, 142.
EXERCISE XXI.— SULPHATES
Barium chlorid or lead acetate give white precipitates, insoluble in dilute acids.
Estimation. — Abderhalden's Handb., 3, 794; 5, 288, 307; Johnston and Adams, 1911,
Jour. Amer. Chem. Soc., 33, 829; volumetric, North, 1914, Amer. Jour. Pharm., 86, 249;
total S. in Urine, Denis, 1910, Jour. Biol. Chem., 8, 401; volumetric, Raiziss and Dubin,
1914, Jour. Biol. Chem., 18, 297; Conjugated, after drugs, Abderhalden, 3, 947, 955.
EXERCISE XXH.— SULPHURIC ACID
Isolation. — Extraction with alcohol; neutralization with XaOH; evaporation; solution
in water.
EXERCISE XXHI.— SULPHLDS
1. Reactions. — They blacken lead acetate. Dilute acids liberate H»S (odor).
2. Detection of H2S in Air. — (a) Blackening of lead acetate paper, (b) Aspirate air
through dilute ammonia containing a few drops of dilute nitroprussid: violet color.
3. Quantitative Estimation: Abderhalden's Handb., 3, 657; in Air: Lehmann; ref.,
Gadamer, 51.
EXERCISE XXIV.— SULPHITES
i Reactions. — (a) Acids liberate sulphur dioxid (odor).
(b) Solutions blacken mercurous nitrate.
(c) Added to Zn and HC1, they develop HjS.
• election of Sulphur Dioxid in Air. — (a) Odor.
(b) Paper impregnated with p<>t;i— him iodate and starch is colored blue.
(c) The air is aspirated through water. This gives the sulphate reactions after addi-
tion of ehlorin-water.
3. Detection of Sulphite in Meat.— (a) Place on start h-iodate pajn-r and n
with dilute sulphuri. a( id: immediate deep blue color (late light blue is insigniii- ant).
(b) S to 25 gm. are subjected to test i (c). The sample is pla. rd in a :o
r flask, and diluted with water if necessary. Some zinc and hydn* hl«>ri. i id are
•itn.du, <d. and the flask tightly stoppered, fixing a strip of lead acetate na|>cr \\ith
the stopper IIS -A ill 1.. i by the rediution <>f the sulphite^, and bla« ken the
absence of sulphite-*, but a Matkening «>uld also be
due to sulphide In this < ase it i- necessary to distil the a. ululated sample in a current
> ( whi, h may be generated directly in the flask by the additi- » <),). The
distillate is re.eived in a standardized solution ..f iodin. \\hi.h is then titrated with sodium
ilphate (also Gadamer, 53).
4. Quantitative Estimation.— Off. Agr. Chem.
EXERCISE XXV.— THIOSULPHATES (HYPOSULPHITES)
'phur di..\id ami pre, ipitatc sulphur. They give a white
precipitate with lead acetate, turning black on heating.
84 A LABORATORY GUIDE IN PHARMACOLOGY
QUESTIONS ON CHAPTER XII
1. How would you test a solution for a carbonate and a bicarbonate?
2. How would you test an antiseptic solution for borate?
3. How would you determine whether an epilepsy mixture contains
bromid?
4. How would you determine whether a tablet contains potassium
chlorate?
5. How would you confirm that a patient is taking iodid?
6. How would you test for free iodin in a vomitus?
CHAPTER XIII
FLAVORS
The mouth should be rinsed after tasting each solution.
EXERCISE I.— SWEETENING AGENTS
Determine the sweetening power of the following drugs as compared
with i per cent, cane-sugar. Start with the strengths given below and dilute,
each time with equal quantities of water, continuing until the taste is less
sweet than that of saccharose. Then try two dilutions between this and
the preceding. Note any qualitative difference in taste.
Each pair of students determine the saccharin; the others will be assigned:
1. Sodium saccharin, o.oi per cent.
2. Glycerin, 10 per cent.
3. Lactose, 10 per cent.
4. Glucose, 10 per cent.
5. Levulose, 10 per cent.
QUESTION
Tabulate the results in multiples of cane-sugar (e. g., saccharin = 300
X cane).
EXERCISE II.— DILUTION
Compare the taste of the following: (a) undiluted; (b) diluted with 10
volumes of water. Taste the weaker solutions first.
Character of taste.
1. Magnesium sulphate, 20 per cent. ) c r
2. KBr (or KI), 5 per cent, f '
3. Sodium salicylate, 10 per cent Mawkish.
4. Chloral,1 10 per cent Acrid.
5. Quinin bisulphate, i per cent Bitter.
6. Saccharin, o.i per cent Sweet and bitter.
QUESTIONS
(a) Tabulate the results, indicating whether the dilution is markedly,
moderately, or scarcely effective in disguising the taste.
(b) For what classes of substances would dilution be effective?
(c) For what classes of substances would it be ineffective?
(d) What qualitative change is there on diluting the saccharin?
1 Taste the strong solution cautiously.
CHAP. XIII FLAVORS 85
EXERCISE m.— COMPARISON OF WATER AND MILK
Compare the taste of solutions i to 5 of Exercise II, diluted with 10
volumes of (a) water, (b) milk.
QUESTIONS
(a) Tabulate the results, indicating whether milk is markedly more
effective than water.
(b) For what classes of substances would milk be especially indicated?
(c) For what classes of substances would it be superfluous?
EXERCISE IV.— COMPARISON OF WATER AND ACACIA
As in Exercise III, using 5 per cent, acacia instead of milk.
QUESTIONS
Analogous to Exercise III.
EXERCISE V (OPTIONAL).— COMPARISON OF WATER AND AROMATIC
WATER
As in Exercise III, using peppermint-water instead of milk.
QUESTIONS
Analogous to Exercise III.
EXERCISE VI (OPTIONAL).— HOT AND COLD SOLUTIONS
As in Exercise III, comparing the diluted solutions cold and hot.
QUESTIONS
Analogous to Exercise III.
EXERCISE VH.— COMPARISON OF WATER AND SYRUP
As in Exercise III, using syrup instead of milk.
QUESTIONS
Analogous to Exercise III.
EXERCISE VIH (OPTIONAL).— CONCENTRATED AND DILUTED SYRUP
Compare the taste of solutions i to 5 of Exercise II diluted with 10 volumes of (a)
syrup; (b) diluted syrup (i : 10).
QUESTIONS
(a) Tabulate the results, indi. ating whether concentrated syrup is markedly more
• than diluted syrup.
(6) For which classes of substances would dilution of the syrup be permissible?
(c) For which not?
EXERCISE IX.-COMPARISON OF SYRUP AND ACID SYRUP
As in Exercise VIII. comparing dilution with (a) simple syrup, (b)
citric acid, i per cent, in >yrup.
QUESTIONS
Analogous to Exercise VIII
86 A LABORATORY GUIDE IN PHARMACOLOGY
EXERCISE X (OPTIONAL).— SYRUP AND SYR. AURANTII CORT.
As in Exercise IX, using orange syrup instead of the citric acid.
QUESTIONS
Analogous to Exercise IX.
EXERCISE XI (OPTIONAL).— SYRUP AND TOLU SYRUP
As in Exercise IX, using Syr. Tolu.
QUESTIONS
Analogous to Exercise IX.
EXERCISE XH.— SYRUP AND GLYCYRRHIZA
As in Exercise IX, using Syr. Glycyrrh.
QUESTIONS
Analogous to Exercise EX.
EXERCISE Xm.— SYRUP AND ELIXIR
As in Exercise IX, using elixir.
QUESTIONS
Analogous .to Exercise IX.
EXERCISE XIV (OPTIONAL).— SYRUP AND COMP. TINCT. GENTIAN
As in Exercise IX, using the Tincture.
QUESTIONS
Analogous to Exercise IX.
EXERCISE XV.— SYRUP AND CO. TR. CARDAMOMI
As in Exercise IX, using the Tincture.
QUESTIONS
Analogous to Exercise IX.
EXERCISE XVI.— SYRUP AND SYR. ERIODICTYON
Analogous to Exercise IX.
EXERCISE XVII.— QUININ
Compare the taste of che following in the order given:
1. Equinin (quinin ethylcarbonate).
2. Quinin adsorbed by Fuller's earth.
3. Quinin tannate.
4. Quinin alkaloid.
5. Quinin sulphate.
QUESTION
Arrange results in order of taste.
CHAP. XIII FLAVORS 87
EXERCISE XVm.— FATTY OILS
Compare the taste of cod-liver oil in the following:
1. Pure.
2. With addition of 0.4 per cent, peppermint oil.
3. With addition of 0.4 per cent, lemon oil.
4. In 50 per cent, emulsion, unfavored. *
QUESTIONS
Record the efficiency hi correcting the oily taste.
EXERCISE XIX.— INSIPID POWDERS
Compare the chalky taste in the following:
1. Pure chalk.
2. Chalk, i; milk-sugar, i.
3. Chalk, i; cane-sugar, i.
4. Chalk, i; sugar, 0.5; cacao, 0.5.
5. Chalk, i; sugar, 0.8; cinnamon, 0.2.
QUESTION
Which is the most effective flavor?
EXERCISE XX.— TASTE OF CATHARTIC SALTS
Compare the taste of the following in 5 per cent, solutions:
1. Magnesium sulphate.
2. Sodium sulphate.
3. Sodium phosphate.
4. Sodium-potassium tartrate.
5. Sodium citrate.
QUESTIONS
(a) Record your results as to degree of disagreeable taste.
(b) Which of the salines would be easiest to take?
(c) Which would be the most difficult?
GENERAL QUESTIONS ON CHAPTER XHI
Which flavors would be suitable and which unsuitable for:
(a) Saline taste (Magn. sulph., KHr, KI)?
(b) Mawkish or alkaline taste (salicylate or bicarbonate)?
(c) Acrid taste (ammonium dilorid or carbonate, chloral)?
(d) Bitter taste (quinin, strychnin, etc.)?
(e) Oily taste?
(/) Chalky taste?
88 A LABORATORY GUIDE IN PHARMACOLOGY
CHAPTER XIV
(DEMONSTRATION) COLORS; (OPTIONAL) DETECTION OF
COLORS IN FOODS, ETC.
(DEMONSTRATION) COLORS
Tabulate the colors produced by the addition of the following colors
to:
(a) Water; (b) sod. bicarb.; (c) I per cent. HCl; (d) 10 per cent, suspen-
sion of chalk.
i. Liq. Carmini,
0.3
i
per cent
«
3
it
2. Tr. Cardam. Co.,
25
ii
So
<f
75
((
3. Tr. Personis,
0.3
n
n
5
ti
4. Fid. Ext. Glycyrrh.,
5
it
15
tt
25
(i
5. Tr. Caramel, N. F.,
0-5
ti
tf
'
u
6. Tr. Hydrastis,
0-3
it
i
It
3
((
7. Tr. Curcumae,
0-3
tt
i
11
3
t(
8. Methylene-blue,
O.OI
ft
O.I
It
9. Chalk with Carmin,
i : 500 (dry).
(OPTIONAL) DETECTION OF COLORS AND PRESERVATIVES IN
FOODS, ETC.
The following table of commonly used preservatives and colors may serve as a guide
to the examiner:
Wines and Grape-juice. — Cochineal; Coal-tar Dyes; Salicylic Acid; Sulphites.
Beer, Cider, Ginger Ale. — Caramel; Salicylic Acid.
Liquors and Vinegar. — Caramel.
Canned Vegetables, Catsups, and Other Sauces, Pickles. — Copper; Saccharin; Sulphite;
Coal-tar Colors (tomatoes); Salicylic and Benzoic Acid; Borate.
Butler. — Coloring; Borax.
Milk and Cream. — Coloring; Formaldehyd; Borax.
Jams and Jellies. — Coloring, Coal-tar and Cochineal; Salicylic and Benzoic Acid;
Saccharin.
Meats.— Coal-tar Colors; Sulphite; Borax; Fluorid.
Syrups. — Saccharin.
Color Standards. — Arny and Ring, 1916, Jour. Ind. Eng. Chem., 8, 309.
EXERCISE I.— COAL-TAR COLORS
Dyeing Test. — Some pure white wool (nun's veiling) is cleaned by boiling for a short
time with very dilute KOH, thoroughly washed, dried, and cut into pieces about 3 by
10 cm. These may be preserved in stoppered bottles.
CHAP, xiv COLORS; DETECTION OF COLORS IN FOODS 89
If the suspected material is liquid, about 100 c.c, are taken. If it is solid or semi-
solid, 5 to 25 gm. are diluted with water to 100 c.c. The liquid is acidulated with 2 to 4 c.c.
of 10 per cent. HC1. A strip of the wool is added and the mixture boiled for five to fifteen
minutes.
If the wool is not appreciably dyed in this time, no coal-tar color is present. If it is
dyed, it is washed in cold water; warmed for a few minutes with very dilute HC1; and
again washed thoroughly. It is then placed in a 2 per cent, ammonia solution, wanning
if necessary. When the cloth is nearly or quite decolorized it is taken out of the liquid,
and this is diluted to 50 c.c., rendered moderately acid with HC1, and another sKp of the
cloth is added. It is then warmed in the water-bath. Natural colors will not give any
appreciable stain in this second dyeing; whereas coal-tar colors and some lichens will give
a well-marked color to the wool.
EXERCISE H.— RED COLORS
1 . NaOH Test. — The origin of red colors may be discovered by the addition of NaOH;
this causes a change to green if the pigment is that of fruits; to blue or purple if it is of
other vegetable origin. Anilin dyes are not changed. For other tests, see La Wall, 1905.
2. Cochineal. — The foods, etc., are first extracted with water if necessary. The
filtered solution is acidulated with HC1 and extracted with amyl alcohol; this is colored
yellow to orange if cochineal is present. The amyl alcohol layer is drawn off and washed
three times with a little water. On adding a very dilute uranium acetate solution, drop
by drop, to the amyl alcohol a characteristic emerald green color appears if cochineal is
present.
EXERCISE HI.— YELLOW COLORS
1. Turmeric (Curcuma). — (a) To some of the i per cent, tincture add a drop of NaOH
(reddish-brown color); then an excess of dilute HC1: yellow color is restored.
(b) Dip some paper which has been dyed with curcuma (Turmeric Paper) into 5 per
cent, boric acid: orange color. Touch it at one place with dilute HC1 and dry: deeper
red. Moisten with ammonia: deep blue. (This serves also as a test for boric acid.)
In applying this test for the discovery of turmeric in solids, these are first extracted
with alcohol.
2. Artificial Coloring Matter in Milk (Leach's Method).— Heat 150 c.c. of the milk
in a porcelain capsule and add enough acetic acid (about 5 c.c.) to curdle. Stir and heat
to near boiling. Gather the curd in one mass on the stirring rod or strain if necessary.
The artificial color will be found in the curd.
is pressed as dry as possible, and macerated for several hours (over nigh;
50 c.c. of ether in a small, tightly corked flask. The ethereal solution is treated according
to (a), the curd by (ft).
(a) Annatto. — This is contained in the ether extract which is decanted and evapor-
ated. The residue is made alkaline with dilute NaOH, warmed, and passed through a
wetted filter. The fat is washed from the filter by a stream of water and the p.r,
dried. If annatto is present the paper has an orange color. A drop of stannou> t hlorid
solution changes this to pink.
- (b) If the curd, after the extraction with ether, is of a purr white color, no artifu ial
dye is present. If it is yellowish or orange this indicates anilin orange; if it is brown.
caramel is suspected. Proceed by (c).
(c) Anilin Orange.— \ lump of the curd is shaken in a test-tube with strong 1 U
it turns pink at once.
(d) Caramel. — If the curd is brown, and anilin orange is absent, the presence of
caramel may be assumed.
(e) I This may be te-ted for dinvtly in the milk.
rtificial Coloring Matter in Butter. <<i) Amutt? and Saffron.— 5 gm. of t
pected butter an- di--olved in 50 « .< . <.t" ether in a white tube and -1 with
ery dilute K<>H \\hi.h miM remain alkaline after M-JUI
rr is drawn olT (without lilterir:
and rnni-trm-d uith . <>n. nil rated ^ulpln:- \ es a blue or \iolet blur,
.uitkly to ureen. and finally to I .tTron dot*-* not nive the interim
(b) 7 shown by extracting the melted butter with alcohol and applying
the ordinary test.
EXERCISE IV.-DETECTION OF CARAMEL IN LIQUORS, ETC.
i (in shaking the sample the foam has a brown color if a considerable proportion ,.f
caramel is present
go A LABORATORY GUIDE IN PHARMACOLOGY
2. Shake some of the fluid with about one-tenth its volume of Fuller's earth for two
or three minutes. Filter, returning the first portions of the nitrate until it runs clear.
If the color of this nitrate is markedly lighter than that of the original fluid this indicates
the presence of caramel. It is always well to make comparative tests with a sample of
known purity.
3. Identification in Flavoring Extracts. — Lichthardt, 1916, Jour. Amer. Phar. Assoc.,
5» 2Q4-
QUESTIONS
(a) What agents and what proportions are suitable for coloring alkaline
liquids pink, red, brown, yellow, blue?
(b) Ditto for acid liquids?
(c) Ditto for neutral liquids?
(d) Is it advisable to color suspensions?
(e) Write a formula for tooth powder consisting of chalk and colored
pink.
CHAPTER XV
CHEMIC STUDY OF THE EXCRETION OF DRUGS IN MAN
Explanatory. — Drugs are excreted mainly by the urine and feces; to a
lesser extent by the saliva, milk, and other secretions. They may also
be demonstrated in effusions; a few pass into the cerebrospinal fluid, etc.
The following experiments relate mainly to the urine, and involve prac-
tice in the application of the tests and the mapping out of the course of the
normal excretion.
The urinary excretion is generally parallel to the absorption, so that it
also gives a fair idea of the absorption of the drugs. Readily absorbable
drugs usually begin to appear in the urine within one-quarter to one-half
hour, reach their maximum concentration in two to four hours, and then
diminish gradually. Many drugs are temporarily stored in the body, so
that traces continue to be excreted for days, weeks, or even months.
Assignment of Experiments. — The exercises should be divided among
the class so that each student takes one of the drugs, collects the urine and
saliva in fractions as directed, tests each of these fractions for the drug, and
by the relative intensity of the reactions maps out a curve of the excretion.
These results will be presented to the next meeting of the class.
The fractions showing the strongest reaction should be furnished to the
instructor, who will distribute them to the remaining members of the class
for practice in the tests. Each student will, therefore, study his own urines
in detail, and also perform all the reactions on the most typical urines of the
other students. It is also useful to have urines of pathologic cases, especi-
ally nephritis, for comparison.
Collection of Urine. — The experiments are assigned and the drugs
, (average doses) are given out at the previous laboratory meeting. Just
before breakfast, on the morning preceding the laboratory exercise, the
subject collects a sample of urine, empties the bladder, and takes the drug.
The urine is collected separately at the end of the following periods after
taking the drug: one-quarter hour; one-half hour; one hour; two hours; four
hours; six hours; ten hours; sixteen hours; twenty-four hours. Bottles will
be provided by the instructor. If the saliva is to be tested the mouth must
be rinsed thoroughly after taking the drug (or this may be taken in a cap-
CHAP. XV CHEMIC STUDY OF THE EXCRETION OF DRUGS IN MAN 91
sule), and a small sample of saliva (5 c.c.) is collected at the end of each of
these periods.
If the drug is to be applied to the skin it should be painted or rubbed on
the inner side of the arm or thigh.
EXERCISE I, A.— EXCRETION OF IODID AFTER ORAL ADMINISTRATION
Test the urine and saliva, according to Chapter XII, Exercise XIII, No.
2, after taking one of the following drugs:
(1) Potassium lodid, 0.3 gm. in water.
(2) Syr. Hydriodic Acid, 5 c.c. in water.
(3) Strontium lodid, 0.3 gm. in water.
(4) Syr. Ferrous lodid, i c.c. in water.
(5) lodalbin (iodin-blood protein compound), 5-grain capsule.
(6) lodipin (10 per cent, iodized sesame oil), 5 c.c. in milk.
(7) Sajodin (calcium mono-iodobehenate), i gm. in milk.
EXERCISE I, B.— EXCRETION OF IODID AFTER DERMAL ADMINISTRA-
TION
Test the urine and saliva according to Chapter XII, Exercise XIII,
No. 2.
(8) KI Ointment. — Rub about i gm. of 10 per cent. KJ ointment (U.
S. P. VIII) into the skin (prolonged friction).
(9) lodin Tincture. — Paint a square inch of the skin with Tr. lodin.
EXERCISE II.— LIBERATION OF IODIN FROM KI BY NITRITES OF THE
SALIVA
(Each pair of students should try this experiment.) Mix equal parts of
i per cent. KI and i per cent. H2SO4, add a little starch paste, place in three
test-tubes, and add to:
(a) Saliva; (b) boiled saliva; (c) water, a and b both turn blue, while c
remains unchanged. Since the reaction is not destroyed by boiling, it can-
not be due to ferments. (It is caused by the presence of nitrites in the saliva ;
the depth of color varies greatly in different individuals.)
EXERCISE HI, A.— EXCRETION OF SALICYL AFTER ORAL ADMINISTRA-
TION
Test urines according to Chapter VII, Exercise IV, No. 3.
1. Sodium Salicylate, i «m. in water.
2. Acetyl-salicylic Acid (aspirin), 0.3 gm. capsule.
3. Methyl Salicylate (oil of wintergreen or birch), 10 drops in capsules.
4. Phenyl Salicylate (salol), 0.3 gm. powder.
5. Sal it 'in, \ mn. powder. (Salicin splits into dextrose and saligenin.
The latter oxidizes into salicylic aldehyd and salicylic acid.)
EXERCISE III, B. EXCRETION OF SALICYL AFTER DERMAL APPLICATION
Test urine according to Chapter VII, Exercise IV, No. 3. Paint the
skin with one of the foll<>\\ b
6. Sodium Salicylate, 5 c.c. of saturated alcoholic solution.
7. Methyl Salicylate, 2 c.c. of 50 per cent, in olive oil.
8. Spirosal (monoglycol salicylate), 2 c.c. of 50 per cent, in oil.
9. Mesotan (nu -th> I oxymethyl salicylate), 2 c.c. of 50 per cent, in oil.
5. tt.-Weighed drugs.
Q2 A LABORATORY GUIDE IN PHARMACOLOGY
EXERCISE IV.— (OPTIONAL) CONVERSION OF BEN7OIC INTO HIPPURIC
ACID
(See Dakin, 1910, Jour. Biol. Chem., 7, 103.)
EXERCISE V.— EXCRETION OF HEXAMETHYLENAMIN AND FORMALDE-
HYD IN URINE AND SALIVA
Test the urine and saliva according to Chapter VIII, Exercise XIII,
No. 4.
Take hexamethylenamin, 0.5 gm. in water.
EXERCISE VI, A.— CONVERSION OF ORGANIC ACIDS INTO CARBONATES
The acid radicles of organic salts are largely oxidized in the body to
carbonates. The acidity of the urine is thereby diminished; or, with large
doses, it may become actually alkaline. Test the reaction of the urines to
litmus.
1. Sodium Acetate, 10 gm. in water.
2. Sodium Citrate, 10 gm. in water.
EXERCISE VI, B.— (OPTIONAL) MEASUREMENT OF HYDROGEN ION
CONCENTRATION
In considering reaction (acidity or alkalinity) it is necessary to differentiate between
total acidity or alkalinity (the entire H or OH available for neutralizing) ; and actual acidity
or alkalinity (the free or dissociated H+ and OH- ions). The potential acidity or alkalinity
(reserve alkali) corresponds to the difference between the two, i. e., to the combined H or
OH.
The total acidity or alkalinity is measured by the ordinary titration methods; the
actual reaction, expressed as H+ ions, may be determined by an electrometric method, by
certain indicators, or by the velocity of certain chemic reactions.
TECHNICAL REFERENCES
Total Acidity of Urine. — Abderhalden's Handb., 5, 283.
Hydrogen Ion Concentration. — Michaelis, Wasserstoffionen Konzentration, Berlin,
1914; Abderhalden's Handb., i, 534; 5,317, 500, 1095; Tigerstedt, 1.2, 186; Palmer and
Henderson, 1913, Arch. Int. Med., 12, 153; Walpole, 1913 (gas-electrode), Bioch. Jour., 7,
410; (litmus), ibid., 7, 260; (indicator chart), ibid., 5, 207; Dreser, 1910 (indicators of effec-
tive alkali), Arch, internat. Pharmacod., 20, 431; McClendon, 1915 (H electrodes), Amer.
Jour. Physiol., 38, 180 (direct reading potentiometer), ibid., 186; Crozier and Harrison,
1915, Surg. Gyn. Obst., Dec., 722; J. H. Long, 1916, Jour. Amer. Chem. Soc., 38, 936.
Preparation of Solitiions. — Abderhalden, 3, 1337.
Alkalinity of Blood. — Abderhalden, 5, 200; Peabody, 1914, Arch. Int. Med., 14, 236;
Heinz, i, 389; Levy, Rowntree, and Marriott, 1915 (dialysis method), Arch. Int. Med.,
16, 389-
EXERCISE VH, A.— EXCRETION OF METHYLENE-BLUE (METHYLTHIONIN
HYDROCHLORID)
Experiment i. — Take 0.15 gm. of methylene-blue in capsule.
The urine has a blue or green color after thirty to fifty minutes. (De-
colorizing under the action of bacteria.)
(a) Boil with a few drops of concentrated HC1: the color becomes pinkish
red; neutralize with NaOH: returns to green.
(b) Add a few drops of NaOH, boil, and add a few drops of i per cent.
glucose1 solution: the color disappears, but reappears on shaking.
(Detection of chromogens in urine, Fleig, 1909, Chem. Abstr., 3, 552.)
1 The urine often contains enough reducing substance to decolorize on heating, even without
the addition of glucose. This may be tried.
CHAP. XV CHEMIC STUDY OF THE EXCRETION OF DRUGS IN MAN 93
Experiment 2. — Take 65 c.c. of 1.5 : 1000 solution of methylene-blue
(=0.1 gm.), previously shaken with 3 gm. of animal charcoal. The urine
is not colored. Explain.
EXERCISE VII, B.— (OPTIONAL) PHENOLSULPHONEPHTHALEIN EXCRE-
TION TEST
The rate of excretion of this substance is used as a test for renal efficiency (Rowntree
and Geraghty, 1910, Jour. Pharmacol., i, 579; 2, 393); i c.c. of a solution containing 6 mg.
of the phthalein is injected deep into the lumbar muscle. The patient is given 300 to
400 c.c. of water about one-half hour before the drug. The urine is collected at inter-
vals. A drop of 25 per cent. NaOH causes a deep red color.
To estimate the excretion quantitatively, NaOH is added to each sample until the
color reaches its maximum. It is then diluted to i liter, filtered, and compared in a
colorimeter with a standard solution (3 mg. per liter).
EXERCISE Vm, A.— EXCRETION OF WATER
The experiment extends over four days.
On the first day the bladder is emptied before breakfast. At breakfast
the usual amount of fluid is taken (measured), and the urine collected and
measured every hour for four hours.
On the second day the same routine is followed, but an additional 500 c.c.
of water is taken at breakfast.
On the third day, as on the first day, with an additional 500 c.c. of milk.
On the fourth day, as on the first day, with an additional 500 c.c. of water
and i gm. of theobromin-sodium salicylate.
QUESTIONS
1. What proportion of the additional water is excreted by urine within
the four hours (a) with water; (b) with milk?
2. How soon does the additional excretion start, when does it reach its
maximum, and when is it completed (a) with water, (b) with milk?
3. Which is the more efficient diuretic? Why?
4. How does theobromin affect the excretion?
EXERCISE Vm, B.— (OPTIONAL) EXCRETION OF SALT
The experiment extends over four days, with the diet as uniform as pra. ti. al. r-pr. ially
in regard to salt. The total urine of each twenty-four hours is collected, and the per-
centage and total quantity of i lilorid is determined. An extra 10 gm. of salt, dissolved
laki-n at the beginning of the second day.
TECHNICAL REFERENCES
Tests of Kidney Function— R. Fitz, 1914, Amcr. Jour. M«l. Sri., 148, 330; Moscnthal.
1916, Jour. Amer. Med. Assoc., 67, 933; Chace and Myers, 1916, ibid., 67, <; I 'iiu-
and LouKh. I«>IM. An h. Int. Med., 17, 570; Flttorescfin, Strain Beii klin. \\
2226; Ur< -i, iQi6,Jo\. Med. Assoc., 66, 415: J°"r
Addis and Watanabe, 1916, Jour. Biol. Ou-m.. .\\. No. 3; La<
ver and Takaya>u. K,M. Doit. An h. Klin. M-d . 101.
HrdinVr ami S. hl.iyrr. [914, Doit. An h. Klin. M<d, n.j. .. "al. i,,i;. An h.
S3-
Tests of Liver Function.— Chesney, Marshall, and Rowntree, l<
Asso< wntrec, Marshall, and Cl rans. Ass**
29, 586; \\ .1 n>-w«>rk. I II II Bul ; 07, 269, 343, 359; Sisson,
\r.h. Int. Mrd . M.SOI; KrumMia. 10; Jmir ;
'in and Inline
Urotn \\ill.ur and Addis, 1913, Trans. Assoc.
I'hys., 28, 617; Abilcrhald.-n. |, 852, B «5, Zcntrbl. Bioch.
Hi. .ph.. 18, 578; Urockrome and other pigments, ibid., 3, 857; 2, 736.
94 A LABORATORY GUIDE IN PHARMACOLOGY
EXERCISE IX.— EXCRETION OF ACETANILID DERIVATIVES
Test urines by the Indophenol Reaction, Chapter VII, Exercise II, No. 5.
1. Acetanilid. — 0.2 gm. as powder.
2. Acetphenetidin (Phenacetin). — 0.3 gm. as powder.
EXERCISE X.— EXCRETION OF ANTIPYRIN
Take 0.3 gm. in water and test urines according to Chapter VII, Exer-
cise III, No. 4.
EXERCISE XI.— SANTONIN URINE
Take 0.05 gm. of santonin as powder, and test urines according to Chap-
ter VI, Exercise II, Nos. 4 and 5, a, b, d.
EXERCISE XH.— EXCRETION OF EMODIN CATHARTICS
Test urines for chrysophanic acid, according to Chapter VI, Exercise
II, No. 5, d. Also note time of cathartic effect; number and character of
stools; griping, etc.
1. F. E. Rhubarb. — i c.c.
2. F. E. Senna. — 2 c.c.
3. F. E. Cascara.— i c.c.
EXERCISE Xin.— EXCRETION OF QUININ
Test urine for alkaloid: acidulate with dilute sulphuric acid and add a
drop of mercuric potassium iodid: precipitate disappears on heating, reap-
pears on cooling.
1. Quinin Sulphate. — 0.2 gm. capsule.
2. Quinin Alkaloid. — 0.2 gm. capsule.
3. Quinin Tannate. — 0.2 gm. capsule.
4. Quinin Ethyl-carbonate (Euquinin). — 0.3 gm. powder.
EXERCISE XIV.— (OPTIONAL) COPAIBA URINE
Take i gm. of copaiba, and test the urines as follows:
(a) Add concentrated HC1: red color, becoming violet on heating. The spectroscope
shows bands in the blue, green, and orange (Quincke, 1883). The reaction is not pro-
duced by all samples of the drug.
(6) Add ammonia: light brown or bluish fluorescence.
(c) Boil: precipitates; add alcohol: dissolves.
(d) Test for sugar: the result is often positive (due to glycuronic acid).
EXERCISE XV.— FORM OF ADMINISTRATION ON ABSORPTION OF
WATER-SOLUBLE DRUGS
Take KI, 0.3 gm., in the following forms, and determine its urinary ex-
cretion according to Chapter XII, Exercise XIII, No. 2.
1. Solution.
2. Powder.
3. Capsules.
4. Pills.
5. Salol-coated Pills.1
6. Glutoid Capsules.
1 Amer. Pharmaceut. Assoc., 57, 94.
CHAP. XVI CHEMIC ANTIDOTES 95
EXERCISE XVI.— FORM OF ADMINISTRATION OF INSOLUBLE ESTERS
Take phenyl salicylate, 0.3 gm., in the following forms, and determine
its urinary excretion according to Chapter VII, Exercise IV, No. 3.
1. Powder.
2. Powder, with 5 parts of chalk.
3. Capsule.
4. Pill.
EXERCISE XVH.— (OPTIONAL) VEHICLE ON ABSORPTION
Take KI, 0.3 gm., in the following vehicles, and determine its urinary excretion accord-
ing to Chapter XII, Exercise XIII, No. 2.
1. Dissolved in glass of water.
2. Dissolved in glass of milk.
3. Dissolved in i ounce of simple syrup.
4. Dissolved in i ounce of thick starch paste.
5. Dissolved in i ounce of 50 per cent, alcohol.
EXERCISE XVni.— (OPTIONAL) STATE OF DIGESTIVE CANAL ON
ABSORPTION
Take KI, 0.3 gm., in water, under the following conditions, and determine its urinary
excretion according to Chapter XII, Exercise XIII, No. 2.
1. One hour before breakfast.
2. Just after breakfast.
3. One hour after breakfast.
4. Two hours after breakfast.
5. Three hours after breakfast.
QUESTIONS ON CHAPTER XV
1. State for each of the drugs used:
(a) When the excretion begins.
(b) When it reaches its maximum.
(c) When it begins to decline.
(d) When it is reduced to traces.
(e) When it is completed.
2. In the exercises in which several combinations of a drug were used,
arrange these in the order of the rapidity of their excretion.
3. Explain why certain combinations are excreted more slowly.
4. With water-soluble drugs, arrange the forms of administration in the
order of absorption, and explain the reasons for the differences.
5. Same as to insoluble esters.
6. What influence has the vehicle on absorption? Explain.
7. How does food inihu-nce absorption? Explain.
8. How could the urine be rendered alkaline without disturbing the
reaction of the stomach? Explain.
CHAPTER XVI
CHEMIC ANTIDOTES
Explanatory.— One of the first objects in r >niniz
is to render the poison insoluble, thereby deb .il^<.rj)ti..n
agent which is used for this purpose must itself be almost harmless, so that
96 A LABORATORY GUIDE IN PHARMACOLOGY
it can be given in unlimited quantity. With this restriction any precipitant
may be used. (It is useful to remember that these precipitants are gener-
ally employed as tests for the substance.) The subject is also simplified by
the fact that the same chemic antidotes are used for all alkaloids.
EXERCISE I.— ANTIDOTES FOR ALKALOIDS
1. Tannin. — (a) To some ^ Per cent- solution of Strychnin Sulphate
add a little infusion of tea: large precipitate. Add to half of this some
alcohol, to the other half some dilute HC1: the precipitates dissolve.
(b) Repeat with ^ per cent. Morphin Sulphate: only a slight precipitate.
(c) Repeat (a) with coffee infusion : only a slight precipitate.
Tannin is an efficient precipitant of some alkaloids, but not of others.
Coffee is less efficient than tea. The precipitates dissolve in alcohol and in
dilute acids.
2. lodin. — To some saturated aqueous Quinin Sulphate add some solu-
tion of iodin in KI: large precipitate. Add some alcohol: the precipitate
dissolves.
3. Permanganate. — To some quinin solution add solution of KMnO4:
brown precipitate. Add alcohol: no solution.
The reactions 2 and 3 apply to all alkaloids, so that these reagents may
be considered universal alkaloidal antidotes.
EXERCISE II.— ANTIDOTES FOR METALS
1. Tannin. — (a) Add some tea to Lead Acetate: large precipitate. Add
to half of this some alcohol: no solution; to the other add dilute HC1: the
tannate is decomposed and lead chlorid is precipitated.
(b) Repeat (a) with HgCl2: very little precipitate.
(c) Repeat (a) and (b) with coffee: results similar to tea.
Some metals are precipitated by tannin, others not. The precipitates
are insoluble in alcohol, somewhat soluble in dilute acids.
Coffee-tannin is also effective, but less than tea.
2. Proteins. — Mix some HgCl2 and albumin solutions: large precipitate.
Practically all metals are precipitated by proteins.
EXERCISE HI.— SPECIAL ANTIDOTES
1. Barium and Sulphates. — To some barium chlorid solution add Na2S(>4
solution: white precipitate.
2. Oxalates and Calcium. — To a solution of potassium oxalate add some
Ca(OH)2: precipitate.
3. Phosphorus and Copper. — Drop a small piece of phosphorus into a
dilute solution of CuSO4: the phosphorus is soon covered with a film of
metallic copper.
EXERCISE IV.— (DEMONSTRATION) BULK OF ACID AND ALKALI
REQUIRED FOR NEUTRALIZATION
i. Neutralization of Sulphuric Acid. — Place about J ounce of concen-
trated sulphuric acid in each of three large beakers; add to (a) Sodium
Bicarbonate; (b) Magnesium Oxid; (c) Sodium Hydroxid (10 per cent.) until
neutral to litmus.
S. M. — Strych. Sulph., ^ per cent.; Morphin Sulph., -^ per cent.; infusion of tea, infusion
of coffee; egg-white solution; phosphorus in small pieces.
CHAP. XVII ADSORPTION BY COLLOIDS
97
2. Neutralization of Sodium Hydroxid. — Place i ounce of 10 per cent.
NaOH in each of two beakers; add to (a) Dilute Acetic Acid; to (6) Dilute
Hydrochloric Acid until neutral to litmus.
QUESTIONS ON CHAPTER XVI
1. Tabulate the chemic antidotes for alkaloids, metals, lead, barium,
oxalates, phosphorus.
2. State your observations; explain the chemic changes.
3. Does the administration of the chemic antidotes suffice for the treat-
ment of poisoning? Why?
4. Which is most effective as a precipitant, tea or coffee?
5. Should alcohol be administered when the chemic antidotes for alka-
loidal poisons are employed? Why?
6. Which would be the best alkali to use against poisoning by acids,
and vice versa? Why?
CHAPTER XVII
ADSORPTION BY COLLOIDS
Explanatory.— Fine solid particles have the property of condensing many
dissolved substances on their surface, and thus removing them from solu-
tion. This effect increases with the surface, and, therefore, with the fine-
ness of the particles. It is especially striking in colloid solutions, in which
the particles are ultramicroscopic, transitional between solution and solid.
This adsorption may be utilized to delay local action and absorption and as
antidote in alkaloidal poisoning. However, this use is limited, since the
dissolved matter is eventually given up in the intestines by simple solution
or by change of reaction.
Adsorption in biochemic analysis, Abderhalden's Handb., 6, 100.
EXERCISE I.— (DEMONSTRATION) ADSORPTION BY INDIFFERENT
SOLIDS
Fill a 6- to lo-inch percolator with dry sand, and tap to park UK- sand.
Pour on to this a i : 100,000 solution of methylene-blue: the solution becomes
decolorized as it passes through the sand.
EXERCISE H.— (DEMONSTRATION) ADSORPTION OF ALKALOIDS, ETC.,
BY CHARCOAL
Experiment i. Strychnin. — Mix 10 c.c. of a i : 1000 solution
nin sulphate with i «ni. of powdered wood charcoal in a flask, and BJ
occasionally during one-half hour. Filter, and compare the filtrate with the
nal solution:
(a) The filtrate is tasteless.
(6) It ^ive> no precipitate with mercuric-potassium iodid.
Freshly calcined animal charcoal acts similarly, and also removes color-
ing malt
Experiment 2. Dyes.— Shake 20 c.c. of a 1.5': 1000 solution of met h
blue with o. i gm . of bone-black : complete dccoloriza t ion should occu r \\ i t h i n
one minute. (Test for quality of charcoal.)
98 A LABORATORY GUIDE IN PHARMACOLOGY
EXERCISE m.— (DEMONSTRATION) ADSORPTION OF ALKALOIDS BY
COLLOIDAL CARBON (CARAMEL)
Proceed as in Exercise II, using about i gm. of caramel (burnt sugar)
in place of the charcoal and leaving for only ten minutes: the nitrate does
not precipitate mercuric-potassium iodid (Sabbatani, 1914, Arch, di Farma-
col., 16, 518).
Purified Caramel. — Berenger, 1912, Amer. Jour. Pharm., 160; Beal and Zoller, 1914,
Jour. Amer. Pharm. Assoc., 3, 495.
EXERCISE IV.— (DEMONSTRATION) ADSORPTION OF ALKALOIDS BY
HYDROUS ALUMINUM SILICATE (FULLER'S EARTH; LLOYD'S
REAGENT; ALKRESTA)
To 10 c.c. of an acid i : 1000 solution of quinin sulphate add about i gm.
of the earth. Shake occasionally during ten minutes. Filter a few drops
and test with Meyer's reagent: negative. Render the mixture alkaline
with ammonia; shake with chloroform; evaporate a few drops of the chloro-
form solution and test with Meyer's reagent: positive.
The earth adsorbs alkaloids from acid or neutral solutions more rapidly
than does bone-black. The adsorbed alkaloid is liberated by alkalies
(Gordin and Kaplan, 1914, Jour. Amer. Pharm. Assoc., 3, 627; Fantus,
1914, Ibid., 3, 657; Rehfeld, ibid., 710; J. U. Lloyd, 1918, ibid., 5, 490).
The earth also precipitates barium chlorid, lead acetate, zinc sulphate, etc.
TECHNICAL REFERENCES
Estimation of Adsorbing Efficiency of Fuller's Earth. — The decolorization of malachite
green is used for this purpose (Fantus, 1915, Jour. Amer. Med. Assoc., 64, 1838).
EXERCISE V.— COLLOIDS ON TASTE
Compare the taste of the following, dissolved in water, with the same
strength solutions in 10 per cent, starch paste:
1. Citric acid, i per cent.
2. Quassia, TV per cent.
3. Quinin bisulphate, T^ per cent.
4. Sugar, 5 per cent.
5. Salt, 3 per cent.
QUESTIONS ON CHAPTER XVTI
1. Define and explain adsorption.
2. What class of substances act as adsorbents?
3. What effect would the combination of an alklaoid with an adsorbent
have on (a) its taste, (b) its systemic action?
4. What substances can be improved in taste by adsorbents?
CHAPTER XVIII
(DEMONSTRATION) SELECTIVE SOLVENTS
The distribution of a substance between two solvents is determined by
chemic and solution affinity and by adsorption. The selective affinity of
drugs for cells is controlled by the same principles.
5. M.— Solutions for Exercise V.
artment
of
CHAP.
XVIII
SELECTIVE SOLVENTS
Pharmacology
EXERCISE
I.— DISTRIBUTION COEFFICIENT
of
I.
20 C.C
Chloroform. — In a 5o-c.c. graduated cylinder place 2c
. of olive oil, and 10 c.c. of chloroform. Shake occa«
c.c. of bitterp to
ifinfl]|y_Hiirinp
fifteen minutes; let stand, and read the volume of the two solutions.
2. Alcohol. — Perform a similar experiment, using alcohol instead of
chloroform.
3. Ether. — Use either instead of chloroform.
The distribution-coefficient = volume of substance dissolved in oil -5-
volume dissolved in water.
EXERCISE II.— ABSTRACTION OF PHENOL BY SOLVENTS
25-c.c. portions of a i per cent, aqueous solution of phenol are shaken
occasionally during fifteen minutes with the following solvents. A little of
the aqueous solution is decanted, and the intensity of the color given by
ferric chlorid is compared with that of the original phenol solution:
1. Kerosene oil.
2. Olive oil.
3. Turpentine.
4. Ether.
EXERCISE m.— VOLUME OF SOLVENTS
Shake lo-c.c. portions of a saturated aqueous solution of iodin with the
following. Compare the depth of color of the iodin hi both layers:
(a) 10 c.c. chloroform.
(ff) 50 c.c. chloroform.
(c) Five successive lo-c.c. portions of chloroform.
EXERCISE IV.— COMPETITION OF SOLVENTS
Add a little dry starch to i : 10,000 solutions of iodin in —
(a) Water.
(b) Alcohol.
(c) Chloroform.
Shake and let settle until clear. Note changes in the color of the starch
and of the solvent.
EXERCISE V.— INTERMEDIARY SOLVENT
Shake some powdered iodin with the following. Compare the depth of
color:
(a) Water.
(b) 25 per cent, alcohol.
EXERCISE VI.— DISTRIBUTION BY CHEMIC AFFINITY
Shake 10 c.c. of a dilute iodin solution in chloroform with the following.
Note the color of the chloroform solution :
(a) Wai
(b) 5 per cent. NaOH.
EXERCISE VH.— (SPECIAL ASSIGNMENT) EVAPORATION OF ANES-
THETIC MIXTURES
e the spc t\ M| a mixture of equal pan- of chloroform and
ether. I-AajKirate one portion (a) spontaneously and another (b) by a bri-k
current of air. Control tin- >p« \ ity when one-quarter, one-half, and
tlmr-quarters have been evaporatt
100 A LABORATORY GUIDE IN PHARMACOLOGY
TECHNICAL REFERENCES
Vital Staining. — L. B. Wilson, 1915, Jour. Lab. Clin. Med., i, 40.
QUESTIONS ON CHAPTER XVIII
1. PARTITION COEFFICIENT OF ANESTHETICS
(a) What is the partition coefficient of chloroform, alcohol, and ether?
(b) The membrane and contents of nerve-cells are rich in lipoids. On the
assumption that the anesthetic action is conditioned on the lipoid content,
what would be the order of efficiency of the chloroform, alcohol, and ether?
Does this conform to the facts?
2. EXTRACTION OF PHENOL BY SOLVENTS
(a) What is the order of solubility of phenol in the fluids?
(b) What bearing has this on the treatment of phenol poisoning?
(c) Would an oily solution of phenol be antiseptic? Why?
3. VOLUME OF SOLVENT
(a) Is a given quantity of solvent more effective if used in one or in
several fractions? Why?
(b) Would it be possible to remove iodin from chloroform by means of
water? Why?
4. COMPETITION OF SOLVENTS
Arrange the solutions in the order of the intensity of the blue color.
Assuming that the blue color is due to solution of iodin in the starch, explain
why the intensity of the starch reaction is unequal in the different solutions.
5. INTERMEDIARY SOLVENT
(a) Why does the solution take up more iodin in the presence of alcohol?
(b) Explain the possible bearing of this observation on the fact that the
activity of a substance (in this case, the coloring power) may be increased
by a second substance that does not itself possess this power (potentiated
synergism; amboceptor group).
6. DISTRIBUTION BY CHEMIC AFFINITY
(a) Why is the iodin removed from chloroform by NaOH and not by
water?
(b) Explain the possible bearing of this observation on the fact that the
pharmacologic activity of a substance may be diminished (or increased) by
the addition of a second substance that may itself be inactive.
7. EVAPORATION OF ANESTHETIC MIXTURES
(a) Does the specific gravity remain constant during the evaporation?
(b) Does the composition remain constant?
(c) In what way would this interfere with the administration of anes-
thetic mixtures?
CHAP. XDC OSMOSIS AND DIFFUSION IOI
CHAPTER XIX
(DEMONSTRATION) OSMOSIS AND DIFFUSION
Explanatory. — The protoplasm of cells takes up water and swells when they are
placed in dilute solutions; while it loses water and shrinks when they are placed in strong
solutions of salts, and indeed, of most soluble substances. This process is called osmosis.
In order that osmosis may occur it is necessary that the two solutions (in this case the
protoplasm and the salt solution) have a different concentration; and that they are
-\ited by a membrane (the cell wall) which is permeable to water, but not to the dis-
solved molecules. A membrane of this kind is called semi permeable. A membrane which
is not quite impermeable to the dissolved molecules but which interposes more resistance
to them than it does to water, may be termed partly semi permeable. Most, if not all,
cell walls belong to the last class; so does parchment. These membranes often possess a
different degree of permeability for different salts.
The molecules of a substance in the state of solution behave precisely like the mole-
cules in a gas (Van't Hoff's Theory), and obey the same laws (Gay-Lussac's, Avogadro's,
Boyle-Mariotte's). They therefore tend to distribute themselves evenly through the
space at their disposal, i. e.y through the solvent. \Yhen they are prevented from doing
so by the interposition of a semi or partly semipermeable membrane, they exert a pres-
sure which is strictly proportional to the number of molecules present in a unit of space,
and independent of the nature of the molecules. This is called the osmotic pressure.
A mol (molecular weight expressed in grams) dissolved in a liter of water exerts the
same pressure as a mol of gas confined in the same space, i. e.t 22.34 atmospheres at o° C.
This osmotic pressure can only be realized under the above conditions — i. e., when two
solutions are separated by a semipermeable membrane. If the two solutions have the
same molecular concentration (mols per liter), they will be under the same osmotic pressure;
they are said to be isotonic. If they are of different concentration, the stronger solution
will be under a higher pressure; it is said to be hyper isotonic; the weaker is hypo-isotonic.
This difference of pressure tends to equalize itself by the passage of the solvent through
the membrane, so as to render the two solutions of equal concentration. This changes
the volume of the solutions: the weaker solution diminishes, the stronger gains, in volume.
This is the explanation of the changes in the volume of the cells.
The law that the osmotic pressure is directly proportional to the molecular concen-
tration holds strictly only for substances like urea, alcohol, sugar, etc. It needs to be
modified for acids, bases, and salts; for in dilute solutions the molecules of these substances
fall apart, the fragments acquiring charges of electricity, and being known as ions (Arrkc-
niits' Hypothesis). The degree of ionization increases with dilution. Kach ion behaves
physically like an entire molecule. A very dilute solution of NaCl therefore exerts twice
the calculated osmotic pressure; sulphuric acid (H-H-SO<) three times; sodium phosphate
(Na-Na-H-PCM four times, etc. (The + and — indicate the nature of the el.
charue whit h is carried by the ion.) The undissociated molecules and tin ions, CJ
in a -olution under given conditions, are tailed collectively mol-ions. It is really the
mol-ions, and not the mols, which determine the osmotic pressure.
The experimental determination of the absolute osmotic pressure is beset with >crious
tedinic difficult! •-. It require.- the » on-truction of a vessel with struth cable
-trench to withstand the high pressure.
ll the nearest approach ; ap . ell i- filled wit! Iphate
and set in a solution of potassium ferrocyanid. The two solution- meet in the pore-, and
cause a pre< ipitatcof the reddish-brown copper fern* \anid, which fututi- •
permeable membrane. Osmomclcrs, thistle shaj>cd tubes closed with pan hment. I>1.
or peritoneal n.. nil in . ertain phy-io|oi:ic experiments; hut they arc only
•emipermcaMe.
:nately. there are other properties of solutions whi. h vary prei isely with the
niiilnular « on, ml ration, and \vhi» h are much more easily determined. Sin h an
l>oilim: point or. ino-f . .m\ enientlv. the fi.
•esses the ' int of the water 1>> e\.i. tl\
'•> A 0.589.
Osmotic Pressure Through Partly Semipermeable Membranes. It is evident that
mnot rea< 'i the thcoreti. level; for -onie of the mole, ulcs \\\\\ , ! the mem-
brane is as fwrmeablr
pressure whatsoever, no matter what tin n will therefore
be hypo-isotonic to a solution the dissolved molecules of which cannot pass
102 A LABORATORY GUIDE IN PHARMACOLOGY
brane. One may therefore see the paradoxic phenomenon of a weaker solution (of a
non-permeating substance) being hyperisotonic to a stronger solution (of a permeating
substance). The law that cqu i molecular solutions (having the same molecular concen-
tration) are isotonic holds therefore only for strictly semipermeable membranes. The
cell membrane of the red blood-corpuscles is strictly semipermeable to most substances.
The corpuscles are therefore isotonic to a 0.9 per cent. NaCl solution, and to equimolecular
solutions of most other substances. Urea and ammonium salts are exceptions; they pene-
trate readily, and their solutions are consequently hypo-isotonic and produce laking.
Many other cells (for instance, those of the kidney) show more numerous peculiarities of
penetration.
Solutions of substances with very large molecules always exert a low osmotic pressure,
since even the strongest solutions must have a low molecular concentration. To this class
belong the colloids — gums, proteins, gelatin, etc.
Osmosis is most conspicuous with the substances of small molecular weight, the
crystalloids. It is most important in the case of salts; the subject of osmosis is therefore
often called SALT-ACTION.
EXERCISE I.— DIFFUSION INTO AGAR
Pour a hot 2 per cent, solution of washed agar into two test-tubes until
they are half filled. Let cool and set. Fill one test-tube with a solution
of copper sulphate or methylene-blue (crystalloid substances); the other
with a solution of Congo-red (a colloid) . Let stand one or two days. The
true solution (methylene-blue) will have diffused through the agar; the
colloid solution (Congo) will present a fairly sharp line of separation. (Some
samples of Congo diffuse freely.)
TECHNICAL REFERENCES .
Diffusion Coefficient. — Tigerstedt, 1.2, 202.
Dialysis. — Abderhalden's Handb., 3, 10, 165; Golodetz, 1913, Zs. physiol. Chem., 86,
3*5-
Collodion Membranes. — Hawk, Physiol. Chem., 30; Abel, 1914 (tubes), Jour. Pharma-
col., 5, 275; Beal, 1914, Jour. Amer. Pharm. Assoc., 3, 499; Lillie, 1907, Soc. Exp. Biol.
Med., 4,^111; Meigs, 1913 (with calcium phosphate), ibid., 10, 129; Schoep, 1911, Zentr.
Bioch. Bioph., n, 377 (glycerin to increase porosity; castor oil for elasticity); Meigs, 1915
(also porous cups, phosphate, and ferrocyanid membranes), Amer. Jour. Physiol., 38, 456.
Capsules, Browne and Soletsky, 1914, Sci., 40, 176.
UUrafiltralion. — Abderhalden's Handb., 5, 1086; Zsigmondi, 1913, Zbl. Bioch. Bioph.,
15, 849; Gaudier, 1912, Chem. Abst., 7, 618.
EXERCISE H.— INCREASE OF VOLUME AND PRESSURE BY OSMOSIS
1. Osmo meters. — Fill the bulb of a thistle tube with syrup. Tie a wet
parchment membrane over the bulb ; immerse in a beaker of water, and note
the height of the liquid in the tube from time to time. Lengthen the tube
with another joint of tubing as necessary. The rise of the liquid shows in-
crease of volume and pressure, the parchment acting as a partly semiperme-
able membrane.
2. (Optional) Egg Experiment. — Remove the shell from the broad pole of an egg with-
out injuring the inner skin. Cement a short glass tube to the narrow end with wax; when
a tight joint has been made, pierce the shell through the tube with a hat-pin. Join an-
other piece of tubing, and stand the egg upright in a beaker of water. The fluid rises,
the egg-skin acting as a partly semipermeable membrane.
TECHNICAL REFERENCES
Direct Determination of Osmotic Pressure. — Abderhalden's Handb., i, 513; Tigerstedt,
1.2, 136.
CHAP. XIX OSMOSIS AND DIFFUSION
103
EXERCISE in.— (SPECIAL ASSIGNMENT)
Osmotic Changes in the Weight of Tissues. — Place the following solu-
tions into evaporating dishes:
a. Water.
b. 5 per cent. NaCl.
c. i per cent. NaCl.
d. Urea, 1.89 per cent.
e. Sodium Citrate, 2.74 per cent.
Of the same freezing-point as i per
cent. NaCl.
of anhydrous.
Cut a fresh dog's or rabbit's kidney into sections about i mm. thick.
Rinse a section in i per cent. NaCl for a moment, dry it superficially with
filter-paper, and weigh; lay it in solution a. Prepare other sections in
the same manner, laying them in the other solutions. Leave in the solu-
tions for half an hour, then again dry and weigh the sections. The weights
will be changed, the sections having absorbed or lost water through osmosis:
(a) Increase of weight — water being strongly hypo-iso tonic.
(b) Decrease of weight — 5 per cent. NaCl being strongly hyperisotonic.
(c) Increase of weight — The protoplasm of the kidney cells is there-
fore hyperisotonic to i per cent. NaCl
(and consequently to blood-serum). It
requires about 1.8 per cent, of NaCl to
keep the weight unchanged.
(d) Increase of weight — much larger than in (c). Consequently,
the kidney cells are easily permeable to
urea.
(e) Decrease of weight — The sodium citrate penetrates less readily
than sodium chlorid.
The experiment illustrates strikingly that the osmotic pressure depends
not only on the molecular concentration, but also on the permeability of the
cell wall, which is different for each substance in the kidney. Urea pene-
trates readily, chlorid less, and citrate still less so.
TECHNICAL REFERENCES
Abderhalden, 3, 542, 547; D. Cohnheim, 1913, Zs. physiol. Chem., 84, 481; Ehrenberg,
1913, Arch. ges. Physiol., 153, i; Hirokawa, 1908, Beitr. chem. Physiol., n, 458.
EXERCISE IV.— PASSAGE OF FLUID BY SOLUTION-AFFINITY
(L'HERMITE EXPERIMENT
In a graduated 5o-c.c. cylinder (stoppered) place, without mixing. 25 c.c.
of chloroform, 3 c.c. of water, and 22 c.c. of ether. Let stand a week and
longer, observing the layers.
EXERCISE V.— "OSMOTIC PRESSURE" BY SOLUTION-AFFINITY
Fill a very thin rubber balloon (condom) with olive oil. Tie a long glass
tube in the opening, and immerse into a cylinder filled with ether. ()!>-
fr-.m time to time. The liquid rises in the tube just as in an osmometer
(W. J. Gies, etc., 1912; Bioch. Bui., 2, 55).
EXERCISE VI.— ALTERATIONS IN MEMBRANE PERMEABII I 1 V
Into lo-i of fresh dog's intestine (ligated at both ends) pl.u e
10 c.c. of 2 percent. NaCl, containing tin- n;imol l.elov.. I
each loop in a large test-tube containing equal amount of water, sufficient to
cover the loop.
104 A LABORATORY GUIDE IN PHARMACOLOGY
Let stand two hours; remove the water, and test it for chlorid with silver
nitrate and nitric acid. Compare the intensity of the Cl reaction in the
dialysates.
(a) 2 per cent. NaCl as control.
The other tubes, each 2 per cent. NaCl, with the following additions:
(b) Lactic Acid, 0.5 per cent.
(c) Sodium Carbonate, 0.5 per cent.
(d) Calcium Nitrate, 0.5 per cent.
(e) Mercuric Chlorid, o.i per cent.
(/) Picric Acid, saturated.
(g) Salicylic Acid, 0.3 per cent.
(h) Magnesium Sulphate, 5 per cent.
(i) Phenol, i per cent.
QUESTIONS
1. Which agents increase the permeability?
2. Which agents diminish the permeability?
3. Suggest explanations.
EXERCISE VIL— IMBIBITION
Lay plates of dry gelatin, about i cm. square, into the following liquids,
and observe after an hour or longer whether they have swollen :
(a) Water.
(b) Oil.
(c) Absolute alcohol.
(d) 50 per cent, alcohol.
(e) 25 per cent, alcohol.
QUESTIONS
(a) Arrange the plates in the order of swelling.
(b) Explain the cause of the differences.
TECHNICAL REFERENCES
Tigerstedt, 1.2, 4, 209; M. Fischer, (Edema, 1910 (Wiley and Sons).
EXERCISE VIII.— CHEMIC CHANGES BY ADSORPTION
Pack a wide glass tube loosely with absorbent cotton. Immerse one end
into a solution of Congo-red, rendered slightly acid. In a few minutes the
cotton immediately above the solution will be colored blue (acid reaction) ;
above this red (neutral or alkaline reaction) ; above this it will be wet but
colorless. The cotton, therefore, adsorbs the acid ions first, then the alkali
ions (E. G. Parker, 1913, Jour. Agric. Res., i, 179).
QUESTIONS
(a) Are all the ions of a salt absorbed equally?
(b) How may this affect chemic processes?
QUESTIONS ON CHAPTER XIX
(a) What causes the molecules of the methylene-blue to move through
the agar?
(b) Why cannot the Congo move in the same way?
(c) Do all kinds of crystalloids diffuse through all kinds of colloids?
(d) What would happen if the passage of the methylene-blue were im-
peded, while water could pass freely?
CHAP. XX DETERMINATION OF MOLECULAR CONCENTRATE
(e) What causes the water to rise in the osmometer or egg?
(/) What would happen if the membrane were impermeab
solvent and solute?
ment
logy
e alike to tka
Toronto
(g) What changes would a semjpermeable cell undergo wlfen laid in (a)
a hypotonic; (b) a hypertonic solution?
(h) Would the change be strictly proportional to the molecular concen-
tration when different solutes are compared? Why?
(i) What is Van't Hoff's explanation of the nature of osmotic pressure?
(j) What other explanation does the L'Hermite and Gies experiments
suggest?
(k) Why does the ether pass into the chloroform and into the oil; and
not vice versa?
(/) Is the permeability of a membrane constant, or may it alter?
(m) Why do the gelatin plates swell in some liquids and not in others?
(n) What are the differences between imbibition and osmosis?
(0) How may adsorption affect chemic reactions?
CHAPTER XX
(OPTIONAL) DETERMINATION OF MOLECULAR CONCENTRATION
Explanatory. — Since osmotic effects depend on molecular concentration,
the determination of this concentration is important. With pure solutions
of non-electrolytes the concentration can be computed from the percentage
of the dissolved substance (gm. per L. divided by molecular weight =
molecular concentration). With electrolytes a correction must be applied
to allow for dissociation.
The total molecular concentration can easily be determined experi-
mentally either by the depression of freezing-point, or by comparing the
effect of the solutions on cells surrounded by a Mini permeable membrane;
for instance, by determining the relative concentration required to produce
laking of red blood-corpuscles.
EXERCISE I, A.— (OPTIONAL) HAMBURGER'S BLOOD-CORPUSCLE
METHOD
Blood-corpuscles are laked \\hen placed in a solution of a certain concentration (about
0.525 per cent. NaCl); the relative concent ration of solutions may. therefore, be deter-
mined by comparing them with a known sodium chlorid solution. This holds true only
if the blood-corpuscles are equally impermeable to the observed substance. It may be
accepted as correct for m«»>t sub-tames, with the notable exceptions of urea and am-
monium -alt-.
•re solutions of NaCl, NaNOj, and Urea, all having the same fn
r c cut. NaCl; 1.535 P**" cen*' NaNOa: i.8q pcj cent. Urea). Set up a scries of clean
;lx>ut 15-c.c. capacity and of equal diameter. \\ii'
'l.i. e in the first 4 c .c. of the NaCl -olution and In the
secoii \ '
C.C. water; fifth, 6 c.c. NaCl and 4 . »NOa
<t urea in th<- .,t lulu- .V: i o drops of
•inatedbl Note the- tub- n \\hi« h there is
ju-t per. cptiblc laking. This will be the same for the , hl.-rid and nitrate, but all the urea
tubes will be laked.
»Th<- part of the experiment should be prepared on the previous day; <>•
results of the experiment being demonstrated.
io6
A LABORATORY GUIDE IN PHARMACOLOGY
EXERCISE I, B.— (OPTIONAL) PLASMOLYSIS
Experiments similar to those with corpuscles may be made with various plant cells.
Haskins (p. 50) used red beet.
TECHNICAL REFERENCES
Blood-corpuscle and Plasmolytic Methods. — Abderhalden's Handb., i, 513; 6, 83;
Tigerstedt, 1.2, 179; Heinz, i, 34, 37.
Osmotic Resistance of Corpuscles. — Stewart, 73. With normal dog's blood, hemolysis
begins with 0.462 per cent. NaCl; and is complete with 0.33 per cent.
Ilcmalocrit. — Stewart, 68; Heinz, i, 39.
Permeability of Cells, determination.— Abderhalden, 3, 545.
EXERCISE H.— (OPTIONAL) DETERMINATION OF FREEZING-POINT
This is done by the Beckmann apparatus (Fig. 5). This consists of a thermometer (g),
with an arbitrary scale (which must be adjusted for each determination, see below)
graduated in o.oi ° C. This is supported by a cork
in a large strong test-tube (e), which may bear a side
piece (/) for the introduction of ice. The cork is per-
forated for a platinum stirrer (h). The test-tube is
supported in a larger tube (d), which acts as an air
jacket, equalizing the temperature. This sits in a jar
(a) of freezing mixture, together with a stirrer (c) and
ordinary thermometer. The principle of the method
consists in overcooling the contents of the test-tube
until ice forms, when the thermometer column sud-
denly rises and comes to a standstill at the correct
freezing-point. The zero point is first controlled by
the standard sodium chlorid solution (10 gm. of dried
salt dissolved in i liter of water, A = 0.589).
Fill the outer jar with a freezing mixture of pounded
ice and salt. This is stirred occasionally throughout
the determination, and kept at about — 5 ° C. by the
addition of salt or ice. Place the standard sodium
chlorid solution in the tube and insert the thermom-
eter, so that the bulb is raised about i cm. above
the bottom of the tube. The level of the solution
should be i to 2 cm. above the bulb. Plunge the
tube directly into the freezing mixture, stirring the
solution constantly. The mercury will be seen to
recede from the reservoir and descend into the stem;
at a certain point it will reverse its motion and as-
cend. Transfer the tube quickly from the mixture to
the jacket-tube, continuing the stirring. When the
columns come to a standstill, take a reading: this is
merely approximate. Remove the tube and stir
until only one or two particles of ice remain.
Plunge for a moment in the freezing mixture, then
into the jacket-tube, and stir until the mercury is
constant. Take a reading with a lens, and repeat
Fig. s.-Beckmann-Heidenhain's melting_and freezing twice. The readings should
apparatus for determining the freez-
ing-point of a solution.
ings must be subtracted,
now be determined.
average. Adding 0.589 to the result gives the zero
point of the thermometer, from which all other read-
The A of defibrinated blood and a sample of urine may
TECHNICAL REFERENCES
Freezing-point Determination. — Abderhalden's Handb., i, 485; 5, 328; 6, 355; 8, 419;
Tigerstedt, 1.2, 140; Heinz, i, 42; Stewart, 492; Haskins, 5i;.Burian and Drucker, 1910
(for 1.5 c.c.), Centr. Physicl., 23, 772; Hartley's Freezing-point Apparatus, Mathews,
Physiol. Chem., p. 201.
CHAP. XX DETERMINATION OF MOLECULAR CONCENTRATION 107
Boiling-point Determination. — Abderhalden, 6, 364; 8, 434.
Melting-point Determination. — Ibid., 8, 419.
'-determination of Molecular Weight. — Barger, 1904, Trans. Chem. Soc., 86, 286;
1915, Abderhalden, 8, i.
DEDUCTIONS FROM DEPRESSIONS OF FREEZING-POINT (A)
The freezing-point can be used for the following calculations :
A
1. The Molecular Concentration = -
1.85
2. The Osmotic Pressure = A X IQ697>8 X d + — ) cm. of mercury or A X *2'34
1.05 273 1.85
t°
X (1 -f — ) atmospheres (t° = temperature in degrees centigrade).
3. The Molecular Weight = 1>8s X per Utef (if no ionization occurs).
4. The Dissociation Coefficient (factor t) = (This factor
gives the ratio on mol-ions to mols. It is^used for deducing the actual freezing-point or
molecular concentration from that which Is calculated on the assumption that no disso-
ciation occurred.)
5. The proportion of ionized molecules (factor a) = -—., i being the factor of the last
-raph; n, the largest number of ions into which the molecules can split (2 for NaCl.
3 for Xa2SO4, etc.).
EXERCISE m.— (OPTIONAL) DETERMINATION OF IONIZATION BY
ELECTRIC CONDUCTIVITY
With a Kohlrausch apparatus determine the conductivity of a NaCl solution. Note
that this is relatively greater the more the solution is diluted, until a constant is finally
reached.
TECHNICAL REFERENCES
Measurement of Conductivity (also in blood, etc.). — Abderhalden, i, 484; Tigerstedt,
1.2, 161; Heinz, 1,46; Stewart, 68.
QUESTIONS ON CHAPTER XX
1. Name three methods of determining the molecular concentration
of a solution of sugar. Would these give concordant result s?
2. How would the results of these methods differ when applied to a solu-
tion of urea? Why?
3. How and why would they differ with NaCl?
4. Explain the Arrhenius hypothesis.
5. What is nu-ant by dissociation coefficient?
6. Work out the following problems:
(fl) What i- tin- molecular concentration of blood-serum, if A = 0.555?
(b) What is its osmotic pressure at 38° C.?
(c) What is the molecular weight of urea, if a 2 per cent, solution =
A 0.62?
(d) What is the dissociation coefficient of a i per cent. NaCl solution
(A 0.589; molecular weight, 58.4)?
(e) What fraction of the molecules is ionized?
108 A LABORATORY GUIDE IN PHARMACOLOGY
CHAPTER XXI
AGGREGATION OF COLLOIDS
Explanatory. — The size of colloid particles is intimately concerned in
their solution or precipitation, surface tension, adsorption, viscosity, and
consistency, etc., and, therefore, in their physiologic properties. This
size, in turn, depends mainly on the electric charges of the particles: increase
of the electric charges tends to make the particles fly apart and thus become
smaller; and vice versa. Electrolytes, therefore, produce important effects
on the properties of colloids.
The affinity of the particles for the solvent is also important. Sus-
pension colloids (for instance, the colloid metals, Prussian blue, etc.) do not
take up the solvent and, therefore, remain discrete. The emulsion colloids,
which comprise most of the proteins and are, therefore, especially impor-
tant in biology, have a strong affinity for water, and adsorb it with con-
siderable force ("imbibition"; for instance, when dry gelatin is laid in water).
When the quantity of solvent is limited the emulsion colloids form "gels";
even solutions have a high viscosity.
TECHNICAL REFERENCES
Properties of Colloids. — Ostwald, Colloidchemie, Dresden, 1911; Abderhalden, i, 508.
Ultramkroscope. — Abderhalden, i, 283.
Refractometer. — Abderhalden, i, 568; 8, 84; Tigerstedt, 1.2, 224.
Surface Tension (see also Viscosity). — Michaelis, Dynamik der Oberflaechen, Dresden,
1909; Tigerstedt, 1.2, 220; Morgan, 1911, Jour. Amer. Chem. Soc., 32, 349.
Viscosity. — Abderhalden, 5, 1358; Tigerstedt, 1.2, 212; Erdmann, 1913, Jour. Biol.
Chem., 14, 141; of blood, Abderhalden, 3, .743; Burton-Opitz, 1911, Jour. Amer. Med.
Assoc., 57, 353; Huerthle, 1900, Arch. ges. Physiol., 82, 415.
Nephelometer. — Marriott, 1913, Jour. Biol. Chem., 16, 290; Richards, 1895, Zs. anorg.
Chem., 8, 269; Richards and Wells, 1904, Amer. Chem. Jour., 31, 235; Bloor, 1915, Jour.
Biol. Chem., 22, 145 (conversion of Duboscq into nephelometer) .
EXERCISE I.— (DEMONSTRATION) VISCOSITY OF SUSPENSION AND
EMULSION COLLOIDS
The viscosity can be judged by the time required for a given column to
run through a certain orifice.
Time the outflow from the same lo-c.c. pipet of:
(a) Water.
(b) Colloidal ferric hydroxid (Merck's dialyzed iron).
(c) 10 per cent, dilution of egg white.
(d) 10 per cent, dilution of egg white containing 1.5 per cent, of Nal.
(e) 10 per cent, dilution of egg white containing 0.6 per cent, of NaCl.
(d and e are m/io solutions; the percentages of m. solutions in this
chapter refer to the grams of anhydrous substance added to 100 c.c. of
water.)
EXERCISE II.— (DEMONSTRATION) GELATINIZATION
Cool a warm 5 per cent, solution of gelatin under the tap: it sets into a
jelly. Heat: it becomes liquid.
Repeat the experiment with a 5 per cent, gelatin solution containing
m/io of the following:
(a) Cane-sugar, 3.4 per cent.
(b) Nal, 1.5 per cent.
(c) Na2SO4, 1.4 per cent., anhydrous.
(b) does not gelatinize on cooling.
CHAP. XXII HEMOLYSIS, CRENATION, AND AGGLUTINATION IOQ
EXERCISE HI.— (OPTIONAL) ELECTROLYTES ON HEAT COAGULATION
Use a 10 per cent, dilution of egg white, which has been dialyzed in a parchment tube
against distilled water for several days. To 5-c.c. portions add 5 c.c. of the following, and
heat to boiling:
(a) Water.
(6) m solution of cane-sugar (34 per cent.).
(c) m solution of NaCl (5.8 per cent.).
Only the last coagulates.
EXERCISE IV.— (OPTIONAL) IONS ON ELECTRONEGATIVE COLLOIDS
To 5-c.c. portions of the dialyzed egg solution (which is electronegative) add 2 c.c.
of each of the following, and let stand several hours if necessary:
(a) m/2o CaCU, 0.55 per cent. (d) 2m KC1, 14.9 per cent.
(6) m/2o MgClj, 0.47 per cent. (e) 2m NaCl, 11.7 per cent.
(c) m/ loo MnCl2, 0.13 per cent. (f) 2m LiCl, 8.5 per cent.
(a), (b), and (c) are precipitated; (d), (e), and (/) not.
EXERCISE V.— (OPTIONAL) IONS ON ELECTROPOSITIVE COLLOIDS
To the dialyzed egg solution add m/5o HC1 (0.07 per cent.) until completely pre-
cipitated; then again the same quantity of HC1. With this solution repeat Exercise 1 \ .
(a), (c), (d), and (e). The reactions are reversed.
Suspension colloids of opposite electric charges also precipitate each other.
EXERCISE VI.— (OPTIONAL) INTERFERENCE OF ELECTROLYTES
To 5-c.c. portions of the dialyzed egg solution add —
(a) 5 c.c. of m KI (16.6 per cent.) + 2 c.c. m/2oMgClj (0.47 percent.): no precipi-
tation.
(b) 5 c.c. of water + 2 c.c. m/20 MgCU (0.47 per cent.): precipitation.
QUESTIONS ON CHAPTER XXI
(a) Describe the characteristic differences between emulsion and suspen-
sion colloids.
(b) Why is the aggregation of colloids affected by salts and not by sugar?
(c) Why are electronegative colloids precipitated by bivalent cathions,
and not by monovalent?
(d) Why is this reversed by the addition of HC1?
(e) What effects have Nal and Na2SO4 on the fluidity and viscosity of
gelatin? Why do these salts differ in their effect?
CHAPTER XXH
HEMOLYSIS, CRENATION, AND AGGLUTINATION OF RED
BLOOD-CORPUSCLES
If the experiments of this chapter have been performed in other courses
they need not be repeated, but they should be read and the questions
answered.
Explanatory.— A. Hemolytii flaking) consists essentially in a soli c cor-
puscles; after a preliminary swelling. irlobin and salts pass fr.mi tin- COfJHttcki
into tin- M-rum (which therefore becomes colored). The stroma can at first be <1
cui-hcd, especially by staining, as colorless "ghosts/' floating in the amber-colored fluid.
These also are eventually dissolved.
110 A LABORATORY GUIDE IN PHARMACOLOGY
Laking agents act by increasing the permeability of the cell envelope. This consists
largely of fatty (lipoid) substances, especially lecithin and cholesterin. All fat solvents —
ether, alkalies, saponin, etc. — therefore produce laking. The bacterial hemolysins prob-
ably act analogous to saponin.
The entrance of water into the cell also causes laking. This occurs when the cell is
laid in water or in any solution of a weaker salt-content than serum. The result is due
to osmosis.
B. Stronger salt solutions, on the other hand, withdraw water from the cell and
shrivel it, producing "crenation."
C. Agglutination consists in the clumping of corpuscles. It is probably due to a
change in the viscidity of the envelope. It may be produced by dilute acid and some other
chemic agents, but is seen most typically with certain toxins, the agglutinins.
EXERCISE I.— TEST-TUBE EXPERIMENTS ON HEMOLYSIS
(Students may work in groups of four.)
Put into 8 perfectly clean and dry test-tubes :
(a) 5 c.c. of 0.9 per cent, sodium chlorid.
(b) 5 c.c. of 0.9 per cent, sodium chlorid containing -^ per cent, of crude
saponin.
(c) 5 c.c. of 0.9 per cent, sodium chlorid containing TV per cent, of crude
saponin which has been digested for an hour at 40° C. with 2 drops of i
per cent, cholesterin solution in ether.
(d) 5 c.c. of 0.9 per cent, sodium chlorid, saturated with ether.
(e) 5 c.c. of 0.9 per cent, sodium chlorid containing i per cent, of urea.
(/) 5 c.c. i per cent. urea.
(g) 5 c-c- 2 Per cent- Na2CO3.
(7i) 5 c.c. of distilled water.
(e and/ must be freshly prepared.)
Add to each tube 2 drops of defibrinated blood and shake. Observe
after half an hour in which tube laking has taken place, as denoted by the
clearness of the mixture or the color of the supernatant fluid.
Isotonic solution of sodium chlorid (a) is indifferent, and does not cause
laking. The addition of sapotoxin (b) dissolves the fatty envelope, and thus
allows laking. If cholesterin (c) is added, th,e sapotoxin is bound and can-
not act on the corpuscles, and there is no laking. (In the body the choles-
terin of the blood acts as a protective in this way.) Ether (d) and other fat
solvents, as* also alkalies (g) also cause laking by dissolving the fatty enve-
lope. Water (ti) injures the corpuscles by removing the salts. Urea (/),
to which the corpuscles are permeable, acts like water. In either case the
addition of salt in isotonic proportion (a and e) prevents laking.
(Optional). — Carmin-fibrin is said to behave to hemolytic agents similarly to blood
(M. H. Fischer, 1909, Koll. Zs., 5, 146).
TECHNICAL REFERENCES
Hemolysis Technic. — Abderhalden's Handb., 5, 24; Fuehner, Nachweiss, 32; Hemoly-
sis and Agglutinin Experiments, Stewart, 70; Antigen, Abderhalden, 3, 1191; Osmotic
resistance of corpuscles, Stewart, 73.
Bio- estimation of Saponin. — Kobert, 1910, Yearbk. Amer. Pharm. Assoc., i, 446.
Antihemolytic Action of Tea Infusions Against Saponin. — This has been proposed as
a test of genuine tea (Maggiora and Ferron; ref., Zentr. Bioch. Bioph., 18, 199).
Sodium Oleate Hemolysis; inhibition by cholesterin; cholesterin — oleate solutions:
0. Klotz and Bothwell, 1915, Soc. exp. Biol. Med., 12, 199.
Blood-corpuscles. — Abderhalden, 5, 143; Kobert, Intox., i, 158; Stroma, Abderhal-
den, 5, 146; Ratio to plasma, ibid., 3, 538; 5, 148; Blood-count, Tigerstedt, 2.5, i; Heinz,
1, 374; Abderhalden, 3, 714. Data in different animals: J. J. Wells and Sutton, 1915,
Amer. Jour. Physiol., 39, 31.
S. M— Materials for Exercise I.
CHAP, xxiii (OPTIONAL) ANTIBODIES in
Platelets.— Abderhalden, 5, 144; 6, 383; Tigerstedt, 2.5, 134.
Experiments on Blood. — Abderhalden, 5, 21.
Drawing of Blood. — Shaffer, 1914, Jour. Biol. Chem., 19, 297; Abderhalden, 3, 1186;
5, 23; 7, 721; Tigerstedt, 1.2, 116.
Determination of Blood Quantity. — Schiirer, 1911, Arch. Exp. Path. Pharm., 66, 171.
EXERCISE H.— MICROSCOPIC CHANGES IN BLOOD-CORPUSCLES
1. Saponin-laking. — Place a small drop of defibrinated blood, diluted with 10 volumes
of 0.9 per cent. NaCl, on a slide under a cover-glass. Examine with the medium power of
the microscope. Add to one edge a drop of 2 per cent, saponin in 0.9 per cent. Nat '1,
strongly tinged with methylene-blue. It will be seen that the corpuscles swell, then lose
their hemoglobin; but the stroma ("ghosts") remains for a considerable time, and can
be discerned faintly by the methylene-blue stain.
2. Water-laking. — Repeat the last experiment, but add water tinged with methylene-
blue in place of the saponin solution: the corpuscles are seen to swell and to lose their
hemoglobin, but more slowly than with the saponin.
3. Amyl Alcohol. — Repeat the experiment, adding amyl alcohol in place of the water,
the cover-glass a little: the corpuscles become agglutinated into small clumps and
then lose their hemoglobin.
4. Cre nation. — To a drop of defibrinated blood under the microscope add a drop of
saturated salt solution: the corpuscles shrivel and become crenated by the abstraction
of water. Similar phenomena can be seen in most cells.
5. Agglutination by Ricin. — On one end of a slide place a rather large drop of 0.9 per
cent. NaCl; on the other end, a similar drop of o.i per cent, ricin in 0.9 per cent. NaC'l.
Add to each a small drop of defibrinated blood, cover, and examine with the microscope
after fifteen minutes: The corpuscles in the ricin solution will be "agglutinated" into
clumps. Many toxins, and the sera of foreign species, have a similar action.
TECHNICAL REFERENCES
Agglutination. — Abderhalden's Handb., 5, 28; Jour. Lab. Clin. Med., i, 56, 1915;
Stewart, 7, 70; Ricin, W. W. Ford, 1913, Centr. Bact., 58, 139.
QUESTIONS ON CHAPTER XXH
(a) Which substances are hemolytic, and which are not?
(b) What would be the results of injecting water rapidly into a vein?
(c) Would the slow intravenous injection of a saturated solution of ether
in 0.9 per cent. NaCl result in hemolysis? Why?
(d) Would a i per cent, solution of glucose produce hemolysis? (The
molecular weight of glucose is 180.)
(e) Would a large dose of urea, taken by mouth, produce laking?
CHAPTER XXIII
(OPTIONAL) ANTIBODIES
The following experiment- ilhi-tratr the main principles; but they need
not be repeated if they have been studied in other courses.
EXERCISE I.— FOREIGN SERUM
Hemolyris by Foreign Serum.— (n) Wash rabbit's corpuscles with 0.9 pet
NaCl, and add -utti ii-nt o.o IKT rent. NaCl to make a 5 per cent, suspen^
- suspension add 0.5 :
add 0.5 p« : \.i( I In. ulmtc at 40° C. for about two hours,
the corpuscles will be found lakcd by the serum.
(b) Make a similar experiment, using dog or ox corpuscles and rabbit scrum: no
ttdng.
112 A LABORATORY GUIDE IN PHARMACOLOGY
2. Destruction of Complement. — Heat some dog or ox serum at 56° C. for one-half
hour. Repeat experiment i (a), using this serum in place of fresh serum: no laking.
Something essential to hemolysis has been destroyed by the heating (viz., complement;.
Save this material for Experiments 3 and 5.
3. Presence of Complement in Rabbit Serum. — To one-half of the complement-free
mixture from Experiment 2 add 0.2 c.c. of rabbit serum; incubate: laking occurs.
4. Removal of Amboceptor. — Centrifugalize 5 c.c. of the 5 per cent, washed rabbit
corpuscles suspension. Pour away the supernatant saline and cool the corpuscles to o° C.
Add 0.5 c.c. of dog or ox serum, also cooled to o° C., and keep at this temperature for one
hour. Centrifugalize rapidly, and separate the serum and corpuscles (keep the corpuscles
for Experiment 6).
To 0.02 c.c. of the 5 per cent, suspension of the original washed rabbit corpuscles add
o.i c.c. of this serum and incubate. Little or no laking occurs because the first corpuscles
have removed the amboceptor from the serum.
5. Mixture of Amboceptor and Complement. — Add o.i c.c. of the serum of Experiment
4 (which contains complement but no amboceptor) to some of the mixture of Experi-
ment 2 (which contains amboceptor but no complement) : laking occurs on incubation.
6. Demonstration of Fixed Amboceptor in Rabbit Corpuscles. — Wash the corpuscles
from Experiment 4 with cooled 0.9 NaCl. To the separated corpuscles add some of the
serum of Experiment 4 (which contains complement but no amboceptor) : laking occurs,
showing that the corpuscles had fixed the amboceptor.
EXERCISE II.— PRODUCTION OF PRECIPITINS AND HEMOLYSINS BY
IMMUNIZATION
Inject intraperitoneally into a rabbitt 5 to 10 c.c. of defibrinated ox (or dog) blood.
Repeat the injection twice, at intervals of six to seven days each. A week or longer
after the last injection obtain some serum from the rabbit. This contains hemolysin for
the blood-corpuscles and precipitin for the serum of the ox (or dog). These are not
present in the serum of normal rabbits.
1. Hemolysin. — Repeat Exercise I, i, using ox and dog corpuscles with the serum of
the treated and of a normal rabbit: laking occurs only with the corpuscles for which the
rabbit has been immunized.
2. Precipitin. — Make a series of dilutions of dog and ox serum with i to 1000 parts
of 0.9 per cent. NaCl. To 0.5 c.c. of these dilutions add 0.2 c.c. of the immunized rabbit
serum, and keep at 40° C.: turbidity, followed by precipitation, occurs in the serum
toward which the rabbit has been immunized.
TECHNICAL REFERENCES
Immunology. — Abderhalden's Handb., 3, 1185; Tigerstedt, 2.1, 48; Zinnser, Hopkins,
and Ottenberg, Laboratory Course in Serum Study; Precipitins, Abderhalden, 3, 1185;
7> 538; Jour. Lab. Clin. Med., 1915, i, 56.
QUESTIONS ON CHAPTER XXIII
(a) How can it be shown that two distinct substances are necessary for
serum-hemolysis?
(b) Why does not the dog serum lake the dog corpuscles?
(c) Why does not the rabbit serum lake the dog corpuscles?
(d) How can a serum be deprived of its complement?
(e) How can one restore the activity of such a serum?
(/) How can the amboceptor be removed from a serum?
(g) What becomes of it?
(h) How can it be shown that it has not been destroyed?
(i) Is the rabbit incapable of manufacturing amboceptors? Why?
(/) How could you show whether the hemolysin and precipitin in Exer-
cise II are identical?
CHAP, xxiv (OPTIONAL) EFFECTS OF DRUGS ON HEMOGLOBIN 113
CHAPTER XXIV
(OPTIONAL) EFFECTS OF DRUGS ON HEMOGLOBIN
These experiments need not be repeated if they have been performed in other courses.
Explanatory. — The. blood pigment, hemoglobin, gives a characteristic absorption
spectrum (Fig. 6). It is easily altered by chemic reagents, with corresponding modifica-
tions in the spectrum. This is sometimes important in diagnosing poisoning.
Red. Orange. Yellow.
Green.
Blue.
A a, £ C
om>.
Fig. 6. — Spectroscopic bands of blood
TECHNICAL REFERENCES
Experiments on Hemoglobin. — Stewart, 74; Tigerstedt, 2.1, 68; Robert, Intox., i, 94,
163, 273; Spectra, Abderhalden's Handb., 6, 389; Heinz, i, 389; Estimation, Abderhalden,
3, 749; Heinz, i, 377; Haldane, 1901, Jour. Physiol., 26, 497; Haessler and Newiomer.
1916, Arch. Int. Med., 17, 806; Kuttncr, 1916, Jour. Amer. Med. Assoc., 66, 1370; (
to/5, Abderhalden, 5, 203; distinction for species, Reichert and Brown, 1908, Soc
Biol. Med., 5, 66; Spectroscopy, Abderhalden, i, 609.
Influence of Sex and Age.- Williamson, 1915, Jour. Amer. Med. Assoc., 65, 302.
Chemic Tests for Blood. — Lenhartz, 183, 271; Kastle, 1909, Hyg. Lab. Bui. 51; Merck's
Rep., 2<, 434; 27, 484; A. L. Holland, 1913, Med. Rec., Oct.; Benmtkm 1 \\Veney,
1909, Zm. Bioch. Bioph., 10, 320; Bordas, ibid., Lyle, Curtman, and Marshall. 1014. Jour.
Biol. (him.. IQ, 445; Phenol phthalein test, Meyer, 1908, rrf., Amer. IMiarrn. Asso,
398; Merck- kt|>, 23, 285; Guaiac Test, Holland, 1907, Jour. Amer. Med. Assoc., 48,
1942; In fetes, Abderhalden's Handb., 5, 394; Dewis, 1907, Host. Med. Surg. Jour., 157,
169.
EXERCISE L— OXYHEMOGLOBIN AND REDUCED HEMOGLOBIN
Use a solution of 4 parts of defibrinated blood1 in 100 parts of water or of ,V per cent.
NaOI I .
1. Oxyhemoglobin. — Place *nmr «>f llu- Milution in a test-tube and examine with the
spectroscope and n<>te the two dark li Hl-.itini: it necessary).
2. Reduced Hemoglobin. Add a few dr. <; ammonium s-.ilphid to the test-
tube: notice the darker « olor and observe the
EXERCISE n.-CARBON MONOXID HEMOGLOBIN
i. Spectroscopic Test. — Pass s- ;i a tul)»- of tin- .lilutrd blood.
lirinhtcr red. Add a few
id: thr double !>an<:
Carbon M<>no\id. whi< h is the prirnipal t»\i< n :•• oal-gas, acts by com-
bining so firmly with hemoglobin tliat it < annul lake u|> ..\\cen Death, therefore,
1 Dog's blood contains on an average 15 per cent, of hemoglobin; beef's blood, 10 per cent.
8
114 A LABORATORY GUIDE IN PHARMACOLOGY
occurs by asphyxiation. The combination is broken up by a great excess of oxygen, so
that recovery is possible with artificial respiration or the inhalation of oxygen.
The skin and mucous membranes are of a bright, cherry red color in carbonic oxid
poisoning; whereas they are blue in ordinary asphyxia.
a. Color Test. — The color of the blood itself is the most certain proof of carbonic
oxid poisoning. The test is performed as follows:
Add a drop of undiluted blood to each of two test-tubes half filled with water.
Pass a stream of coal-gas through one of the tubes, and note that the color changes from
amber to carmin. In suspected poisoning, a drop of blood is drawn from the finger, diluted
as in the above, and compared with the control tube. The depth of the red color permits
an approximate estimate of the degree to which the hemoglobin is saturated with CO.
3. Chemic Tests. — These depend for the most part on the addition of substances
that change the color of oxyhemoglobin but not of CO-hemoglobin.
(a) NaOH: To a i : 20 dilution of blood add an equal volume of 30 per cent. NaOH:
CO blood remains light red; normal blood changes to dirty brown.
(6) Hydrogen Sulphid. — To a i : 50 dilution of blood add an equal volume of satur-
ated HjS water: CO blood shows no change; normal blood turns dirty green.
(c) Ferrocyanid. — Mix 10 c.c. of blood with 15 c.c. of 20 per cent, potassium ferro-
cyanid and 2 c.c. of 30 per cent, acetic acid: CO blood remains red; normal blood changes.
(d) Tannin. — Shake a i : 5 dilution of blood with 3 volumes of i per cent, tannin:
CO blood is carmin red; normal blood turns gray.
4. Demonstration of CO in Air. — Shake 2 to 5 c.c. of diluted blood (just sufficient to
give spectrum) in a liter flask containing the suspected air; or aspirate 10 liters of the air
through diluted blood. Examine the blood for CO-hemoglobin, as in the preceding ex-
periments.
TECHNICAL REFERENCES
Proof in Blood. — Gadamer, 43; Sand, 1914, ref., Jour. Amer. Med. Assoc., 63, 1890;
Estimation, Abderhalden, 3, 637; Brunck, 1912, Chem. Abstr., 7, 747.
Preparation of CO. — Abderhalden, 3, 735; Work with Gases, Abderhalden, i, 215,
230; 5, 1027; 8, 437.
EXERCISE in.— METHEMOGLOBIN
1. Formation of Methemoglobin. — Put some of the diluted blood (about 15 c.c.)
into a series of six test-tubes. Add the reagents mentioned below, and note changes in
color and spectrum at once. If none appear, place in a water-bath at 40° C. and observe
every half-hour:
(1) 25 drops saturated KC1O3.
(2) 25 drops 5 per cent. Pot. ferricyanid.
3) 25 drops 10 per cent. NaNXV
4) 25 drops i per cent. KMnO4.
5) 25 drops Phenylhydrazin.
(6) 25 drops 10 per cent. Pyrogallol (Methemoglobin spectrum and precipitate of
Hemogallol).
Methemoglobin has a rather brown color and shows a sharp band in the red, closely
resembling acid hematin (see Fig. 6).
To one of the test-tubes which shows a good methemoglobin band, add a little am-
monium sulphid: the reduction occurs comparatively slowly, and more of the reagent is
required.
Explanatory. — Methemoglobin is a peculiar modification of oxyhemoglobin. It
differs from the latter in being less readily reduced. The conversion of any considerable
proportion of the blood pigment into methemoglobin therefore leads to asphyxia, char-
acterized by intense cyanosis. This conversion takes place even more readily in the
body than in the test-tube; the chlorate and the coal-tar products are especially apt to
produce the effect in living mammals, while they act sluggishly on shed blood.
The conversion of hemoglobin into methemoglobin can be effected by: Oxidizing
agents (i, 2, 4), reducing agents (3, 6), coal-tar derivatives (5). The rapidity of the
conversion varies considerably: in 2, 3, and 4 it is almost instantaneous; in i it may require
several hours; the others are intermediate. The results are somewhat different in the
intact mammals, ClOa and the coal-tar products being quite active; (5) may also show the
band of reduced hemoglobin.
2. Cyan-hemoglobin. — Add a drop of 2 per cent, hydrocyanic acid to some of the
diluted blood and to some methemoglobin solution. The first shows no change. In the
second, the color brightens and the spectrum changes so as to resemble reduced hemo-
globin (see Fig. 6). This reaction may be used as a test for hydrocyanic acid or for
methemoglobin.
CHAP. XXV CHEMIC EFFECTS OF CORROSIVES AND IRRITANTS 115
This peculiar combination of cyan and hemoglobin does not occur normally during
life, since the blood does not contain methemoglpbin. The latter may be formed after
death, especially in ecchymotic areas; and the bright red color of these spots is a charac-
teristic feature of cyanid poisoning.
EXERCISE IV.— HEMATIN
The blood in the vessels does not show acid or alkali hematin even in severe poison-
ing, but they may be discovered locally; e. g., in the vomit.
i . Alkali Hematin. — Add a few drops of sodium hydrate to the diluted blood. The
color deepens; the spectrum changes to a broad, diffuse band.
j. Acid Hematin. — Add a little dilute acid to the diluted blood: the color becomes
brownish, and some precipitation may occur. The spectrum shows a sharp line in the
red.
3. Hemochromogen. — Add yellow ammonium sulphid to the hematin solution:
the spectrum shows two bands in the green, the left much stronger.
TECHNICAL REFERENCES
Hematin, Abderhalden's Handb., 2, 617.
EXERCISE V.— HEMATOPORPHYRIN
Add a few drops of blood to sufficient concentrated sulphuric acid to be transparent:
the spectrum shows two bands in the orange and yellow.
Hematoporphyrin does not contain iron. It occurs in the urine after sulphonal
poisoning and can be extracted with amyl alcohol.
TECHNICAL REFERENCES
Determination in Urine. — Abderhalden's Handb., 3, 861.
QUESTIONS ON CHAPTER XXIV
(a) Make a diagram of the spectra of all the compounds studied.
(6) How would you distinguish between oxyhemoglobin and reduced hemoglobin?
(c) Describe three characteristic tests for CO in blood.
(</) How would you distinguish between acid hematin and methemoglobin?
CHAPTER XXV
CHEMIC EFFECTS OF CORROSIVES AND IRRITANTS
Explanatory. — All substances which enter directly into chemic combina-
with proteins produce local effects. /. *., they act at tin- place where
they are applied. The action results in inflammation; these substances are
therefore irritants; if the action is at all violent the celK are killed. If
the combination of the reagent and protoplasm is fluid the tissue is disso!
Thi> i- termed corrosion or fiintcriztilion. If, on the other hand, the a.
is mild and the product insoluble, the effect i- </v/rw^-;//. ;'. <\. mu<
membranes are constricted and puckered, and the phenomena of a pre-
mmation are lessened. These precipitates aU«» -rive to stop
the lumen of bleeding vessels and arc. there-
in! to know whether the art inn
Milt- in pre< inflation «»r solution. Thi- mav be studied on isolated protein-.
It must be remembered, however, that tin- efferts depend greatly upon tin-
concentration of the reagent: the precipitates often redissolve in an excess
of the reagent or of the protein.
Il6 A LABORATORY GUIDE IN PHARMACOLOGY
The color of the compounds is often important in diagnosis.
The application of the corrosives to excised tissues shows that these
influence the effect; the skin is generally more resistant than the softer struc-
tures. The tissues also illustrate the stains and the penetration of the
corrosion.
(Students may work in groups of four.)
TECHNICAL REFERENCES
Investigation of Irritants, Heinz, i, 255.
EXERCISE I.— PROTEINS (EGG-ALBUMEN)
Place in each of twelve test-tubes i inch of a solution of egg-albumen
(the white of 2 eggs to 200 c.c. of water, strained). Add the following
reagents (the usual solutions), drop by drop:
(i) HgCl2; (2) AgNO3; (3) CuSO4; (4) Fe2Cl6 (tincture); (5) Lead Ace-
tate; (6) H2SO4; (7) HC1; (8) HNO3; (9) NaOH; (TO) Carbolic Acid (strong);
(n) Alcohol; (12) Tannin (6, 7, 8, 10, and n: full strength).
A white precipitate is given by HgCl2, AgNO3, Pb(C2H302)2, H2SO4,
HC1, CeH/), C2H6O, and Tannin; a greenish-white precipitate by CuSO4;
a yellowish-brown precipitate by Fe^; a yellow precipitate by HNO3.
NaOH gives no precipitate.
Excess of the reagent, redissolves the precipitate in the case of acids, but
not with the other precipitants. (The reagents which redissolve the pre-
cipitate are apt to penetrate more deeply into tissues.)
QUESTIONS
(a) Tabulate the results as to: Precipitate, its firmness or absence; color
changes; resolution cf the precipitate in the reagent.
(b) Which of the reagents would tend to penetrate deepest?
(c) Which would tend to be superficial?
(d) Which would tend to be corrosive ; and which astringent?
(e) Which cause characteristic color changes?
EXERCISE H.— DEFIBRINATED BLOOD
Place about 2 c.c. of defibrinated blood in twelve test-tubes, and add
the reagents as in Exercise I.
A black or brown clot is formed with Fe2Cl6, H2SO4, and HNO3; a brown
or dark precipitate with CuSO4, HC1, and NaOH; a pink or light red precipi-
tate with C6H6O, C2H6O, and Tannin; a gray precipitate with HgCl2, PbAc2,
and AgNO3. Excess of the reagent redissolves the precipitate with a
brownish-red color in the case of acids, and with a garnet color in the case
of soda. The others do not redissolve. (The color is due to acid hematin in
the case of acid, to alkali hematin in the case of NaOH.)
QUESTIONS
(a) Tabulate the results as to: Precipitation and its firmness; color
changes; solution in excess of the reagent.
(b) Which of these agents would tend to be hemostatic?
(c) Which would give ucoffee-grounds" vomit?
S. M. — Egg-albumen.
CHAP. XXV CHEMIC EFFECTS OF CORROSIVES AND IRRITANTS 117
EXERCISE m.— CORROSION OF SKIN
Place bits of fresh mammalian skin into test-tubes containing concen-
trated H2SO4, HC1, HNO3, NaOH, and CeHeO. Leave for fifteen minutes,
rinse in water, and note the effect on the hair and on the epithelial and con-
nective-tissue surfaces.
With the acids, the epithelial surface becomes first white, hard, and some-
what shrunken. With more prolonged action it is gradually softened.
With HC1 it remains white; HNO3, light yellow; H2SO4, brownish. C6HeO
causes a more pronounced shrinking, puckering, and hardening, without
subsequent softening. NaOH softens. The hair is softened and dissolved
by the NaOH, more slowly by the acids. It is not affected by CeHeO.
The connective tissue is rendered softer and transparent and is finally dis-
solved by the NaOH and the acids, and stained as the epithelium. The
carbolic acid affects it as it does the epithelium.
QUESTIONS
(a) Tabulate the results for epithelium, connective-tissue surface, and
hair as to: softening or hardening; shrivelling; color changes.
(b) Which of the agents could corrode the skin?
(c) Which would cause extensive destruction?
(d) Which would act as depilatories?
(e) Which give characteristic stains?
EXERCISE IV.— CORROSION OF MUCOUS MEMBRANES
Slit open a piece of fresh dog's intestine, 3 inches long, and flatten it,
epithelial surface up. With a glass rod apply a drop of the reagents used in
Exercise III. Observe during fifteen minutes. Note the character, color,
and depth of the effect. See whether the epithelium detaches more readily.
The acids first turn the epithelium white and hard, but soon softer and
darker. The underlying tissues appear white and hard, as if cooked. The
epithelium is readily detached. The action extends deepest with HNOs.
This gives a yellow tinge to the stain. H2SO4 gives a brownish color.
C«H«O acts as it does on the skin; its effect extends deeply. NaOH first
softens the tissues and then renders them gelatinous. The epithelium
scrapes off very readily.
QUESTIONS
(a) Tabulate the results, both for the epithelium and underlying tissue,
as to: hardening or softening; detachment of epithelium; depth of corn)-
color changes.
(b) Arrange the reagents in the order of their liability to produce perfora-
•
(c) Which would show characteristic stains at autopsy?
EXERCISE V.— CORROSION OF MUSCLE
Place bits of mu>clr into the reagents used in Exercise III, and ol>
during iit'tccn minutes. KiiiM- in \\atrr and note appearance and con-
sistency.
H2SO4and HC1 soften the muscle without -\\< -II in^ it the «»lor becomes
a deeper red; the muscle then gradually disintegrates, dissolving entirely,
S. M.— Dcfibrinated blood; skin.
Il8 A LABORATORY GUIDE IN PHARMACOLOGY
with a garnet color, in the case of the H2SO4. In HNOs the muscle shrinks
and hardens, the color changing to yellow or brown, with partial solution.
In CeHeO the muscle is bleached, shrinks, and becomes hard, assuming a
"cooked" appearance. NaOH causes the muscle to become red and
swollen; the outer layers soften, become gelatinous, and dissolve to a red
solution.
QUESTIONS
(a) Tabulate the results as to: hardening or softening; swelling; solu-
tion; color changes.
(b) What is the cause of the "cooked" appearance?
EXERCISE VI.— (OPTIONAL) COAGULATION OF MUSCLE
Tease a bit of fresh frog's muscle in normal saline on a slide, examine with the lower
power of the microscope; add a drop of concentrated H2SO4 and observe the results.
Repeat with the other reagents mentioned in Exercise III, and also with i per cent, caffein.
The acids cause the fibers to shrivel and to become contorted; they turn granular and
opaque, the striations are lost, and gradual solution occurs. C6H6O acts at first like the
acids, but there is no solution. NaOH causes the fibers to swell, to become transparent,
and to gradually dissolve. Caffein produces a granular opacity.
QUESTION
Tabulate the changes in the structure of the muscle-fibers.
EXERCISE VII.— STAINS ON HUMAN SKIN
The observations made on excised tissues apply also to 'the human skin.
The stains may be removed in the manner indicated in the experiment.
1. Apply to the intact skin of the forearm a drop of concentrated nitric
acid. Wash off as soon as there is itching. An intensely yellow stain de-
velops. Apply a drop of ammonia: the stain turns to an orange brown
(xanthoproteic reaction) . It is very lasting and wears off only as the skin
is desquamated.
2. Apply to another place a drop of saturated picric acid: yellow stain.
Apply ammonia: the stain is removed.
3. Apply concentrated sulphuric acid and hydrochloric acid to different
places; wash off as in i: there is no stain, but redness.
4. Apply strong alcoholic solution of methylene-blue; wash off after one
hour. The stain is not removed by water, but by rubbing with dilute
ammonia.
5. Apply a drop of tincture of iodin to two places, leave for five minutes:
mahogany stain which cannot be washed off. Apply ammonia to one of
the spots and sodium thiosulphate solution to the other: the stains disappear.
QUESTIONS
(a) Tabulate the results as to color of the stains; time of appearance;
persistence.
(b) Which reagents give a yellow stain? How can they be distinguished?
(c) How can an iodin stain be removed? How can a silver stain be
removed?
S. M— Dog's intestine; muscle.
CHAP. XXVI PHYSIOLOGIC EFFECTS OF IRRITANTS IIQ
CHAPTER XX\ I
PHYSIOLOGIC EFFECTS OF IRRITANTS
Explanatory. — The tissues respond to irritants by the phenomena of
inflammation. Four successive stages may be recognized in the skin: (i)
Rubefaction, or reddening with pain and itching; (2) Vesication, or blister-
ing; (3) Pustulation, the formation of isolated pustules; and (4) Corrosion,
or destruction of tissue. The degree of the action depends on the nature
and the concentration of the irritant. The rapidity of action is also vari-
able. Chloroform and turpentine, for instance, act quickly, but scarcely
progress beyond rubefaction; cantharides, croton oil, and antimony act
slowly, but progress, the first to vesication, the last two to pustulation. A
quick action is generally associated with volatility. Vesication demands
that the irritant should remain in the skin sufficiently long to produce an
inflammatory exudate under the impermeable stratum corneum. Pustu-
lants have a specific chemotactic power on leukocytes.
Mucous membranes show only rubefaction and corrosion, the ana-
tomic conditions being unsuitable for vesication or pustulation. The mouth,
however, is an exception, for vesication may occur here. Irritation of
mucous membranes is also characterized by catarrh — i. e., increased excre-
tion of mucus. This is diminished by astringents. These also cause puck-
ering.
The treatment of irritation consists in the removal of the irritant and the
application of fats, glycerin, or mucilage. This may be studied on carbolic
acid.
TECHNICAL REFERENCES
Determination and Comparison of Local Toxicity of Chemic Compounds. — Cooper,
1915, Proc. Amer. Soc. Biol. Chem., 3, 19.
EXERCISE I.— IRRITATION OF SKIN
1. Rubefaction.— (a) Rub a little chloroform on the arm. Note the
burning and reddening.
(b) (Optional). — Apply a mustard paper to the chest until tingling occurs. Note
the sensation and the redden ing.
2. (Optional) Vesication. — Apply some cerate of cantharidrs (or o.i mg. of cantharidin
in a drop of oil) to the arm. Cover with adhesive plaster and leave for six hours: a bli-trr
forms. Wash off tin- rxirx of tin- • •antharix with alcohol.
Demonstration) Urticaria. — A few drops of //ijtom/n, i : 1000, are rubbed
ously into tin- >kin <>r. brtu-r. applied to a non-bleeding scratch; or 0.5 t
injn-ted hypodermii ally. This produces a typical urticaria ( r.ppin:
Woch., No. 23). Similar wheals are raised by morphin (i : 100) or its esters. Epi-
ncphrin (i : 1000) produces intense blanching and "goose-flesh." Vcratrin (i : 10,000),
produces intense shooting pain
4. (Optional) Pustulation.— Apply a drop of a 25 per cent, solution of crolon oil in
cottonseed oil to the skin of the arm; a pustular eruption is developed after some time.
QUESTIONS
(a) Tabulate the effects as to inun>ity, onset, and durati
(ft) Which of the drugs are classed as rubefacicnts, vesicants, and
pustulants?
(c) Can chloroform or mustard produce vesication?
120 A LABORATORY GUIDE IN PHARMACOLOGY
EXERCISE II.— IRRITANTS ON MUCOUS MEMBRANES
1. Shake a bottle containing soap-bark and smell it: sneezing.
2. Place a drop of ten times diluted tincture of aconite on the tip of the
tongue: persistent tingling sensation.
3. Place a drop of Tr. lodin on inner surface of lip: blister.
4. Observe the astringent taste of a 5 per cent, solution of alum, of Tr.
.Ferri Chlor., and of Tannin.
5. (Optional). — Snuff a very little mixture of i part of veratrin and 500 parts of starch:,
sneezing and all the phenomena of acute coryza.
6. (Optional) Quantitative Estimation of Aconite by Squibb's Taste Method. —
Details, Ford, Ford and Wine, 1915, Amer. Jour. Pharm., 87, 489.
QUESTIONS
(a) Tabulate the observations.
(b) Name some sternutatories (drugs producing sneezing).
(c) Would iodin produce a blister in the stomach? Why?
(d) Does the aconite produce inflammation? What is the difference?
(e) Name some astringents.
(/) For what conditions could they be useful?
EXERCISE in.— (DEMONSTRATION) TREATMENT OF IRRITATION
(PHENOL)
1. Effects of Solvents. — Arrange five small beakers in a circle so that
the fingers can be plunged into them simultaneously. Fill these beakers
with 5 per cent, carbolic acid in (a) water; (b) 25 per cent, alcohol; (c) 25
per cent, glycerin; (d) turpentine; (e) cottonseed oil.
Insert the five fingers of the left hand, one in each solution; keep in for
five minutes, withdraw, and note the blanching and wrinkling, the tingling
(felt especially on pressing the fingers against a table), and the anesthesia.
The effects (especially the blanching) are greatest in the water; much less
in the alcohol and glycerin ; practically absent in the oil.
Rinse the finger which has been in the watery solution in a liberal quan-
tity of water: the blanching persists. Rinse it in 95 per cent, alcohol:
the blanching disappears.
2. (Optional) Phenol Burns. — Dip the tips of two fingers into undiluted liquefied
carbolic acid for one minute. Very little burning is felt, but the skin becomes white. Now
rinse the one finger in water, the other in 25 per cent, alcohol. The latter removes the
blanching, but not the sensory phenomena. It is effective against the superficial actions,
but not against those which are situated more deeply. Glycerin, oil, or turpentine act
like alcohol. Rinse the other finger in the alcohol. There will be some subsequent rough-
ening and chapping of the skin.
3. (Demonstration) Solvents on Precipitation. — Pour J inch of undi-
luted egg-white into two test-tubes; pour over this (without mixing) in
(a) an equal volume of 5 per cent, phenol in water; to (b) in oil; (a) pre-
cipitates at once, (b) very slowly. The phenol, being very soluble in oil,
does not pass into the watery egg-white.
Explanatory. — The reagents (b) to (e) of Experiment i are all better
solvents for carbolic acid than is the skin; they consequently lessen the
penetration of the phenol and hence its effects (Experiment 3). (These
solutions of phenol are therefore also much less efficient as antiseptics than
are watery solutions.) The glycerin and cottonseed oil act in addition in
5. M— Soap-bark; Tr. Aconite, diluted i : 10.
S. M. — Egg-white; 5 per cent, phenol in oil.
CHAP. XXVHI ANTISEPTICS 121
virtue of their viscidity (*. e., as emollients), hindering the access of new layers
of the solution to the skin. This makes them more effective in the treatment
of carbolic burns; but, on the other hand, it hinders the washing off of the
phenol. Lavage of the stomach with 10 per cent, alcohol is the best local
treatment in internal carbolic acid poisoning. For burns on the skin the
surface should be rinsed with the dilute alcohol and then dressed with glyc-
erin or oil. This treatment does not lessen the effects of the carbolic acid
which has been already absorbed (except that still present in the superficial
layers).
QUESTIONS
(a) Tabulate the results.
(b) What would be the proper treatment of phenol burns?
(c) How could these facts be utilized in the treatment of internal phenol
poisoning?
CHAPTER XXVII
(OPTIONAL) CATHARTICS ON MAN
Personal experience with the effects of the common cathartics is very
useful to the physician. Students are, therefore, advised to try the follow-
ing drugs at weekly intervals or as occasion arises, and to report their results
as to time of effect; color, consistence, size and number of stools; griping,
etc. A set of the cathartics will be furnished on application.
LAXATIVES
A loin, 0.15 gm. at bedtime.
Calomel, 0.15 gm. at bedtime.
Cascara, Arom. Fldext., 5 c.c. at bedtime.
Castor Oil, 5 c.c. at bedtime.
Epsom Salt, 5 gm. in glass of water before breakfast.
Petrolatum Liquid, i oz. at bedtime.
Phenol phthalein, o.i gm. at bedtime.
Podophyllum Resin, o.oi gm. at bedtime.
Rhubarb, i gm. at bedtime.
Senn-i, $ urm. at bedtime.
CATHARTICS
Castor Oil, tablespoon before breakfast.
Comp. Jalap Powder, 2 gm. before breakfast.
Epsom Salt, 15 gm. in half a ulass <>i water before breakfast.
Jalap, i gm. before breakfast.
(II \PTER XXYIII
ANTISEPTICS
The relative etVu :• eiit types of antiseptics under actual
working conditions is fairly well illustrated by the following cxpci m
122 A LABORATORY GUIDE IN PHARMACOLOGY
TECHNICAL REFERENCES
Standardization of Antiseptics. — Anderson and McClintic, 1912, Hyg. Bui. No. 82;
Abderhalden's Handb., 5, 9; Heinz, i, 128.
Potassium Tdlurite— Use as indicator of bacterial life, etc., W. E. King and Davis,
1914, Amer. Jour. Publ. H., 4, 917.
Bacterial Cultures and Media. — Abderhalden, 3, 1212; 5, 584.
EXERCISE I.— (SPECIAL ASSIGNMENT) URINARY ANTISEPTICS
Empty the bladder before breakfast, and save the urine. Take one of
the following drugs (which are assigned to different numbers of the class).
Collect the urine at the end of one, two to three, and six to eight hours
after the administration.
Divide the samples into three parts. One is left at its natural reaction;
the second is rendered slightly acid with HC1; the third slightly alkaline
with sodium carbonate.
Incubate the different samples (including the control urine) and ob-
serve after twelve to twenty-four hours for bacterial turbidity and am-
moniacal odor. If these are absent, continue the incubation, examining
daily.
1. Hexamethylenamin 0.5 gm.
2. Sod. Salicylate i.o gm.
3. Sod. Benzoate i.o gm.
4. Creosote 0.3 gm.
5. Methylene-blue ...» 0.2 gm.
6. Boric Acid 0.5 gm.
7. Santal Oil 0.5 gm.
QUESTIONS
Tabulate the results, arranging the drugs in the order of their efficiency,
and grouping them according to whether they are fully effective, moderately
effective, or ineffective.
EXERCISE H.— (SPECIAL ASSIGNMENT) INTESTINAL ANTISEPTICS
Mince a mixture of equal parts of fresh pancreas and duodenum and
mix with a double volume of water. Place equal quantities (about 15 c.c.)
in a series of test-tubes. Add to each 0.15 gm. of the respective drugs.
Stopper the tubes and incubate at about 40° C. Observe daily, noting
the presence and intensity of putrefactive odor.
1. Control.
2. Bismuth Subcarbonate.
3. Calcium Carbonate.
4. Calomel.
5. Charcoal.
6. Creosote.
7. Glutol (Formaldehyd gelatin).
8. Guaiacol Carbonate.
9. Salol.
10. Sod. Phenolsulphonate.
11. Sod. Salicylate.
12. Tannin.
13. Thymol.
CHAP. XXVIII ANTISEPTICS 123
QUESTIONS
Tabulate the results, arranging the drugs in the order of efficiency, and
grouping them according to those which prevent putrefaction, those
which retard it, and those which are ineffective.
EXERCISE m.— (DEMONSTRATION) CALOMEL ON BILE
Place in an incubator some bile in which a knife-point of calomel has been
added, and another sample without this addition, for control. The color
changes first in the latter sample.
EXERCISE IV.— (SPECIAL ASSIGNMENT) WOUND ANTISEPTICS (DUST-
ING-POWDERS)
Place 15 c.c. of fresh defibrinated blood in a series of test-tubes. Add
to each 0.15 gm. of the respective drugs. Stopper the tubes and incubate
at about 40° C. Observe daily, noting the odor. (Laking and the changes
of color are also interesting.)
1. Control.
2. Acetanilid.
3. Betanaphthol.
4. Bismuth Betanaphtholate (Orphol).
5. Bismuth Subnitrate.
6. Boric Acid.
7. Calcium Carbonate.
8. Charcoal.
9. Glutol.
10. lodoform.
11. Tannin.
12. Thymol Diiodid (Aristol).
13. Zinc Oxid.
QUESTIONS
Tabulate trie results, arranging the drugs in the order of efficiency, and
iping them according to those which prevent putrefaction completely;
almost completely; those which merely delay, and those which are inactive.
EXERCISE V.— (SPECIAL ASSIGNMENT) DRYING POWDERS
Mix i c.c. of defibrinated blood with i gm. of the powders in small
dishes and note consistence.
1. Control.
2. Bismuth Subnitrate.
3. Boric Acid.
4. Calcium Carbonate.
5. Charcoal.
6. Kaolin.
7. Starch.
8. Talc.
9. Tannin.
10. Xiiu Oxid.
QUESTION
Tabulate in the order of their i-ilu icncy as absorbents (for wound secre-
tions, etc.).
124 A LABORATORY GUIDE IN PHARMACOLOGY
EXERCISE VI.— (SPECIAL ASSIGNMENT) PENETRATION OF ANTISEPTICS
Place 5 c.c. of each of the following antiseptics in lo-cm. ligated loops of
fresh intestine of rabbit or cat. Be sure that there is no leak. Rinse the
segments in water and place in test-tubes, each with 10 c.c. of water.
At the end of twenty-four hours pour off the water and test for the anti-
septics.
In testing, compare with the original solution diluted 100 times, and, if
necessary, 50, 25, and 10 times.
Note what fraction of the antiseptic has passed through the intestine,
assuming that the antiseptic would have been diluted four times if it had
diffused equally through the fluid.
Reference
Antiseptic solutions. Tests. Chapter. Exercise.
Phenol, 5 per cent Ferric Chlorid. VII I
Compound Cresol Solution, 2 per cent Ferric Chlorid. VII I
Salicylic Acid, saturated Ferric Chlorid. VII IV
Mercuric Chlorid, i : 1000 Ammon. Sulphid. IX XI
KI to solution of precipitate.
Silver Nitrate, i : 1000 Ammon. Sulphid. IX XIII
Tr. lodin Starch Paste. XII XIV
Formaldehyd, i : 5000 Jorissen. VIII XII
Alcohol, 70 per cent Chromate. VIII I
TECHNICAL REFERENCES
Penetration of Antiseptics. — Cheyne's Method, ref., Keilty and Packer, 1915, Jour.
Amer. Med. Assoc., 64, 2123; Kendall and Edwards, 1911, Jour. Infect. Dis., 8, 250.
QUESTIONS
Tabulate the results in the order of penetration, grouping them accord-
ing to those which penetrate readily, with difficulty, and not at all.
CHAPTER XXIX
EFFECTS OF DRUGS ON FERMENTS
Ferments are greatly influenced by conditions, and thus by chemic
substances. However, the effects of drugs are not easily studied under
natural conditions, and unless these are reproduced in detail the results have
little value. They are not of great practical importance, since the drugs,
under natural conditions, do not remain in sufficiently lengthy contact with
the ferments to exert much effect. A few of the reactions, however, are of
special interest.
Students may work in groups of four.
TECHNICAL REFERENCES
Experiments with Ferments.— Robert, Intox., i, 149; Tigerstedt, 2.2, 54; Preparation,
Abderhalden, 3, i; Recognition and Estimation, ibid., 3, 16; quantitative, 24; viscosity,
Feldsteiner and Weyl, 1910, Soc. Exp. Biol. Med., 7, 61; isolation from bacteria, etc., Abder-
halden, 3, 1254; Abderhalden test, Abderhalden, 6, 223.
Trikresol as antiseptic, Graves and Kober, 1914, Jour. Amer. Chem. Soc., 36, 751.
CHAP. XXTX EFFECTS OF DRUGS ON FERMENTS 125
EXERCISE I.— COAGULATION OF MILK
In a series of tubes place 5 c.c. of milk, 5 drops of rennin, and 5 c.c. of
the following reagents; incubate for fifteen to thirty minutes, and note the
occurrence and character of the coagulum:
1. Water.
2. Barley decoction (10 per cent, pearl barley).
3. Pancreatin, o.i per cent.
4. Formaldehyd, o.i per cent.
5. Sodium Citrate, i per cent.
QUESTIONS
(a) Describe the results.
(b) State what practical use could be made of them.
TECHNICAL REFERENCES
Milk Analysis. — Abderhalden, 5, 421; 7, 170.
Preparation of Rennin. — Ibid., 3, 10; Estimation, Hammarsten, 1914, Zs. physiol.
Chem., 92, 119; Casein Estimation, Arny and Pratt, 1906, Amer. Jour. Pharm., 78, 121;
Pasteurized Milk, microscopic stain, Frost, 1915, Jour. Amer. Med. Assoc., 64, 821.
EXERCISE H.— (OPTIONAL) COAGULATION OF BLOOD
Run 10 c.c. of blood from the artery of a living animal into test-tubes containing
2.5 c.c. of the following reagents. Incubate at 40° C, and observe the rapidity and the
firmness of the coagulation:
1. 0.9 per cent. NaCl (control).
2. Ammonium Oxalate, i per cent, in 0.9 per cent. NaCl.
3. Sod. Citrate, 5 per cent, in 0.9 per cent. NaCl.
4. Sod. Fluorid, 1.2 per cent.
lagnesium Sulphate, saturated.
6. HCN, 0.5 per cent, in 0.9 per cent. NaCl.
7. Formaldehyd, i per cent, in 0.9 per cent. NaCl.
8. Leech-head Extract in 0.9 per cent. NaCl.
9. Brain Extract (Kephalin) solution.
QUESTIONS
(a) Record the results.
(6) State the mechanism by which each of the agents hinders coagulation.
TECHNICAL REFERENCES
Experiments on Blood Coagulation. — Stewart, 62; Heinz, i, 386; Abderhalden, 5,
223; Robert. Intnx., i, 158; Coagulation Time, Cannon and Mcndenhall, 1914, Amer.
Jour. I'hy-iol., 34, 215; Hiu-rkiT, lou. Anh. «i-s. Physiol., 149,318.
'-.bin. — Howell, 1913, Amer. Jour. Physiol., 32, 264; Abderhalden, 5, 273.
Antithrombin. — Howell, 1914, Arch. Int. Med., 13, 76; test in blood, Hess, 1915, Jour.
Exp. M»d. .1, \To. 4; Minot and Denny, ioin. Anh. Int. Mrd., 17. 101.
I'ibrinogen. — Whipple, 1914. Amrr. jour. Physiol., 33, 50; Abderhaldrn. 5. 853,
Examination of Blood. — Alxlrrhaldi-n, 3, 742; 5, 155; Total Analysis, ibid., 5, 209.
Dry Residue— Ibid., 5, 155; Ash. il>id.. 159; Specific Gravity. il»i«l.. 3, 742; Lenhart/..
126.
Blood-serum. — Abderhalden, 5, 142; Separation from riot, Sakaguchi, 1912, Zentr.
Bioch. Ilioph., 13, 757-
Serum Proteins. — Refnctomettt detenninatta !>i-rt-«m. Jour. Hi<>l. ciu-m..
22, 233; 33, 325; Tranter and K..\\r, 1915, Jour. Amrr. Mrd. Ass.,, . (,;, 1432; E. ReisS,
Mrd. und Kindh., 10. 531; 1915, D. Anh. Klin. V 175.
Blood Plasma. — Obtaining, Abderhalden, 5, 139, 257, 262, 268.
Calculation of Total Blood in Body.— Abderhalden, 3, 748; Tigerstedt, 2.4, 308;
Drcycr and Ray, 1910, Roy. Soc., 82 B, 545; SchUrcr, 1911, Arch. Exp. Path. Pi
66, i
126 A LABORATORY GUIDE IN PHARMACOLOGY
Average Count, etc., for Dogs, Musser and Krumbhaar, 1914, Fol. Hemat, 18; for
various animals, J. J. Wells and Sutton, 1915, Amer. Jour. Physiol., 39, 31.
Collection of Body Fluids. — Tigerstedt, 1.2, 113.
Bleeding of Rabbits. — Cut edge of back of ear with razor. Put point of split writing
pen into vein in cardiac direction (Zinsser).
EXERCISE m.— (DEMONSTRATION) TRANSFORMATION OF SULPHUR
INTO SULPfflDS
Sulphur owes its irritant action on the skin and intestines to its gradual
transformation into sulphids. This is effected, at least in part, by the
proteins. It is not affected by heat, so that ferments are probably not in-
volved. Throw some pieces of fresh intestine into 20 c.c. of boiling water.
Strain into a small flask. Neutralize. Add a pinch of washed sulphur.
Stopper, suspending a piece of lead acetate paper from the stopper. In
another similar flask place some water, sulphur, and lead paper. In a third
flask place some intestine and boiling water, with lead acetate paper, for
control. Observe that after a time the paper in the intestine and sulphur
flask becomes blackened through the evolution of sulphuretted hydrogen.
(Other proteins give the same result. The experiment is not always suc-
cessful.)
EXERCISE IV.— (DEMONSTRATION) OXIDASE
Guaiac resin assumes a blue color when oxidized. This oxidation occurs
even when the resin is suspended in plain water, but very slowly. It is
greatly accelerated by oxidizing ferments (oxidases) , which are present hi all
living protoplasm. One may use diluted defibrinated blood, or potato
peelings, or fresh lettuce leaves pounded with sand and water and strained.
These are placed in test-tubes, with a drop of fresh guaiac tincture (U. S. P.).
The poison solutions are then added and the depth, of the blue color noted
from time to time. Prussic acid is especially effective in retarding this oxi-
dation. Caffein hastens it somewhat. It is very greatly accelerated by
hydrogen dioxid.
Place into a series of test-tubes equal quantities of potato pulp (peelings
rubbed with water and strained). Add an equal quantity of the reagents
and 20 drops of fresh Tr. Guaiac. Let stand and note the development of
the blue color:
1. Water (control).
2. HCN, i per cent.
3. Quinin Hydrochlorid, 2 per cent.
QUESTIONS
(a) Report the results, arranging them in the order of 'interference.
(b) In the light of these results, what would be the probable actions of
these agents on metabolism?
TECHNICAL REFERENCES
Preparation of Oxidases and Catalases. — Abderhalden's Handb., 3, 42; Measurement in
Plant-juices, Bunzel, 1914, Jour. Biol. Chem., 17, 409; Measurement of oxidation velocity,
Abderhalden, 8, 21; of CO2 production velocity, ibid., 8, 38.
Respiration of Excised Tissues. — Abderhalden's Handb., 3, 444, 451, 460; Batelli and
Stern, 1908, Arch, intern. Pharmacod., 18, 217.
Guaiac as Reagent. — Schaer, 1913, Pharm. Ztg., 63, 328, obtained the best results with
resin extracted from guaiac wood by chloroform; next came the natural resin; and finally
the alcoholic extract of guaiac wood.
CHAP. XXX MONOCELLULAR ORGANISMS AND LEUKOCYTES 127
EXERCISE V.— DIGESTIVE AND SIMILAR FERMENTS
TECHNICAL REFERENCES
Diastase, Preparation, Abderhalden, 3, 387; quantitative, ibid., 6, 231; infeces, ibid.,
5, 404; in feces and urine, T. R. Brown, 1914, Trans. Assoc. Amer. Phys., 29, 547.
Saliva, Examination, Abderhalden, 3, 257.
Invertin, Preparation, ibid., 3, 7, 389.
Pepsin, Preparation and Intimation, Abderhalden, 3, 8; Givens, 1915 (Modified Rose
method), U. S. Hyg. Lab. Bui. 101, 71.
Papain, Standardization, Heyl, 1914, Amer. Jour. Pharm., 86, 542.
Trypsin, Preparation and Estimation, Abderhalden, 3, 9; Long and Barton, 1914,
Jour. Amer. ('hem. Soc., 36, 2151; in feces, Abderhalden, 5, 397; in gastric juice, \Y II.
Spencer, 1015, Jour. Biol. Chem., 21, 165; Pancreatic Juice, ibid., 6, 488.
Erepsin, infeces, ibid., 5, 404.
Secretin, Preparation and Tests, Abderhalden, 3, 205, 418; 6, 487; 7, 65; Launoy
and Oechslin, 1913, Zentr. Bioch. Bioph., 15, 82.
Lipase, Preparation, from liver, Abderhalden, 3, 403.
Castor beans, Taylor on Fermentation, 258; Falk and Sugiura, 1915, Jour. Amer.
Chem. Soc., 37, 217; blood, Whipple, J. H. H. Bui. 24, 357; Quantitative, Abderhalden,
3, 220, 223; in blood, etc., ibid., 8, 301.
Urease, Van Slyke and Cullen, 1914, Jour. Amer. Med. Assoc., 62, 1558.
Xnclease, Nephelometry, Kober and Graves, 1914.
Tissue Juice, Wicchowski's Method, Abderhalden, 3, 282; Beitr. Chem. Physiol., 9,
232, 247.
Buchner Press, Abderhalden, 3, 2.
Autolysis, Alxierhalden's Handb., 3, 433; 5, 1259; antiseptics on, Court, 1915, ref.,
Zentr. Bioch. Bioph., 18, 190; alcohol on, Wells and Caldwell, 1914, Jour. Biol. Chem.,
i9> 57-
Metabolism of surviving organs, ibid., 3, 358; 5, 1215; perfusion, Tigerstedt, 1.4, 51.
Isolation of proteolytic ferments, from Liver and organs, Abderhalden, 3, 407; from
plants, ibid., 413.
Digestion Products, Collection and Analysis, Abderhalden, 6, 458.
Proteolytic Digestion Products, Isolation and Determination, Abderhalden, 3, 227.
Proteoses, Abderhalden, 2, 533; 6, 506; isolation, ibid., 3, 239; silk peptone, ibid.,
5, 578.
Polypeptids, ibid., 2, 529, 545.
Cleavage Products, ibid., 2, 470.
Amino-acids, ibid., 5, ion, 6, 276; Determination in urine, ibid., 3, 810; 5, 309; quanti-
tative, ibid., 2, 470, 510, 559; 3, 1346; in blood, ibid., 5, 190; gasometric, ibid., 5, 995.
Leucin, Determination, ibid., 3, 810; 5, 357.
Tyrosin, Determination, ibid., 3, 810; 5, 357; Folin and Denis, 1912, Jour. Biol. Chem.,
12, 245.
Tryptophan, Isolation, Abderhalden, 3, 246; Cancer Test, Weinstein, 1910, Jour. Amer.
Med. Assoc., 55, 1085.
Gastric Contents, Lenhartz, 254.
Protein Hydrolysis, Comparison of methods, Harding and MacLean, 1916, Proc.
Amer. Soc. Biol. Chem., 3, 15.
CHAPTER XXX
MONOCELLULAR ORGANISMS AND LEUKOCYTES
Explanatory. Poisons arc divided into two groups: (i) Those which
kill all f«.rm> of liviim tissue to which they may l»e applied: and i l) th<»i-
which act selectively, i. e., which have a much stronger action on some ii>
than on others. The first are called general protoplasmic poisons; t In-
second, muscle-nerve poisons.*
1 Their action is not necessarily restricted to muscular and nervous tissue, as the name would
imply. It may also be exerted on gland-cells, etc. The feature of the classification
is that the action is selective.
128 A LABORATORY GUIDE IN PHARMACOLOGY
The general protoplasmic poisons are again subdivided into those which
act also on dead proteins — the corrosives — and those which act exclusively
on living cells — protoplasmic poisons in the restricted sense.
The effects of general protoplasmic poisons are studied most conveniently
on monocellular organisms.
The boundary between the general protoplasmic poisons and the muscle-
nerve poisons is not sharply defined. Many of the typically selective
poisons, such as strychnin, are toxic to all tissues when they are used in
sufficient concentration. The protoplasmic poisons also show some special-
ization. Quinin, for instance, kills ameboid cells much more readily than
it does bacteria. All protoplasmic poisons, however, are to some extent
bactericidal; and all antiseptics can be counted in this group. The estima-
tion of the antiseptic power of protoplasmic poisons belongs to the domain
of bacteriology.
EXERCISE I.— (DEMONSTRATION) YEAST FERMENTATION
Rub a cake of compressed yeast with 100 c.c. of glucose solution. Meas-
ure portions of 10 c.c. into a series of test-tubes and add 10 c.c. of the follow-
ing reagents. Transfer them to fermentation tubes (such as are used in
the fermentation test for sugar). Incubate at about 30° C. for one to two
hours and compare the amount of gas evolved.
1. Water (control).
2. Quinin Hydrochlorid, 0.5 per cent.
3. Strychnin Sulphate, 0.5 per cent.
4. Sod. Fluorid., o.i per cent.
5. HCN, o.i per cent.
6. Sod. Salicylate, 0.05 per cent.
QUESTION
Report the results, arranging them in the order of interference.
TECHNICAL REFERENCES
Experiments with Yeast, Fuehner, 16; Robert, Intox., i, 152; Measurement of Yeast
Fermentation, Abderhalden, 8, 42; Preparation of Zymase, Abderhalden, 3, 393.
EXERCISE II.— (OPTIONAL) PROTOZOA
Macerate a little hay in water for several days until infusoria are developed. Place
a drop of the infusion on a slide and note with the microscope the movements of the
infusoria. Place a drop on each of four slides; add to slide (a) a drop of $ per cent.
quinin; (b) \ per cent, cocain; (c) % per cent, strychnin; (d) & per cent. HgCl2; (e)
caffein, i : 700; (/) NaOH, i : 4000. Cover with cover-glasses (interposing a hair to
prevent pressure) and examine at once, and then every ten minutes. The HgCk kills
the infusoria at once, fixing them in their original elongated shape. The others act
much more slowly; the movements become more sluggish, and finally the infusoria con-
tract to round balls and die. The caffein and alkali cause characteristic structural
changes.
The quinin kills first, then the cocain, and last the strychnin. The observations need
not be continued after the animals in the cocain have died.
QUESTION
Record the results, arranging them in the order of toxicity.
mei
CHAP. XXXI ANTHELMINTICS AND INSECTICIDES I2Q
iversit^
TECHNICAL NOTES
Protoplasmic Poison:. — Heinz, i, 192.
Protozoa. — Abderhalden, 5, 18; Tigerstedt, 1.2, i; Robert, Intox., i, 150.
Amebic Dysentery, Propagation. — Sellards and Baetjer, 1914, ref., Jour.
Assoc., 63, 1789.
t>anosonifs. — Abderhalden, 5, 1371.
Syphilis, Rabbits.— Jacobi, 99.
Tissue Cultures. — Abderhalden, 5, 836; 6, 519; Smyth, 1914, Jour. Amer. Mol.
Assoc., 62, 1377; R. A. Lambert, 1916, Proc. Soc. Exp. Biol. Med., 13, 100; Rous and
Jones, 1916, Suspensions of living cells, ibid., 13, 73.
Transplantation of Organs. — Abderhalden, 5, 828; Tigerstedt, 2.4, 336.
EXERCISE HI.— (OPTIONAL) QUININ ON EMIGRATION OF LEUKOCYTES
Dispose a frog for the observation of the mesenteric circulation. Apply some i per
cent, solution of quinin hydrochloric to a limited space. Observe the effect. Place an
unpoisoned portion in the field and inject i or 2 c.c. of the quinin solution in the dorsal
lymph-sac. Continue the observation for one-half hour, if necessary. (See illustration
in Text-book.)
TECHNICAL NOTES
Experiments on Leukocytes. — Robert, Intox., i, 156; Tigerstedt, 2.5, 104; Obtaining
from blood, Abderhalden, 5, 144; Zinsser, Hopkins, and Ottenberg, "Serum Study," 166;
Glycogenin, ibid., 5, 207; Leukocyte count, dogs, Musser and Krumbhaar, 1914, Fol. He-
matol., 1 8.
Emigration, Ikeda, 1916, Jour. Pharmacol., 8, 101.
Phagocytosis, in vivo, F. C. Mann, 1916, Jour. Amer. Med. Assoc., 67, 174; in vitro,
H. J. Hamburger, Brit. Med. Jour., Jan., 1916.
Opsonic Index. — Zinsser, Hopkins, and Ottenberg, "Serum Study," 168.
Chemolaxis. — Ibid., 5, 1286; Ruchlaedew, 1910, Zs. Biol., 54, 533.
Light, of lions, Abderhalden, 7, 587; determination of intensity, ibid., 6, 180.
Fluorescence, Methods. — Ibid., 3, 1171; experiments on animals, ibid., 5, 563; in
toxicologic analysis, Gadamer, 358.
Radio-activity.— Abderhalden, 7, 788.
Toronto
CHAPTER XXXI
ANTHELMINTICS AND INSECTICIDES
The efficiency of the worm-remedies can be studied outside of the body.
EXERCISE I.— (DEMONSTRATION) ASCARIS
These occur in the pig and may be obtained from the slaughter-house.
They keep alive for several days in a solution containing i per cent. \
and o.i per cent, sodium carbonate (Bunge's solution) if the temperature
is kept constant at 37° to 38° C. At this temperature they are in constant
movement. The action of the principal vermifuge, santonin, is not to kill
the worm, hut to increase its movements in the endeavor to escape from
the santonin solution (v. Schroeder, 1895, Arch. e\p. Path. Pharm.. m. JOG).
I. Place the worms in beakers containing the reagents dissolved in the
Bunge solution, at ^7 (
(a) Men uric Chlorid, o.i per <
(b) Santonin, to saturation.
(c) Chenopodium Oil, i : 5000.
i m a spiral glass tube with Bunge solution at 37° C. Add the v
and keep at thi> temperature. When they have assumed a fairly constant
130 A LABORATORY GUIDE IN PHARMACOLOGY
position in the tube, add a concentrated solution of sodium santoninate to
the open end of the tube: the worms generally move away as the santonin
diffuses into the solution (W. Straub).
QUESTION
Describe the results.
EXERCISE II.— (DEMONSTRATION) ASPIDIUM
This is used for tapeworms, but its activity is tested most conveniently
on the common rain worms.
i. Triturate i gm. of Oleoresin of Malefern with 2 gm. of calcined mag-
nesia until dry. Mix with 10 c.c. of water, let stand a day, decant, and filter.
With a syringe and fine needle inject o.i c.c. of the solution into a large
rain worm, just back of the clitellum. Keep the worm in a little water in a
Petri dish, and observe from time to time: the segments around the injec-
tion swell and flatten. In three to four hours they liquefy.
TECHNICAL REFERENCES
Estimation of Efficiency of Anthelmintics. — Fuehner, 43; Bruening, 1906, Zs. exp.
Path., 3, 564; S. Yagi, 1914, Zs. Exp. Med., 3, 64; Effect of Anthelmintics on rain worms,
leeches, and Ascaris, Trendelenburg, 1915, Arch. exp. Path. Pharm., 79, 190.
EXERCISE III.— (OPTIONAL) INSECTICIDES
A method for determining the activity of fluid insecticides is described by Houghton
and Hamilton, Mich. Acad. Sci., 1909; for fumigants, by McClintock, Hamilton, and
Lowe, Jour. Amer. Publ. H. Assoc., April, 1911.
TECHNICAL REFERENCES
Experiments on Insects. — Fuehner, 50.
Experiments on Invertebrates. — Tigerstedt, 1.2, 69; Robert Intox., i, 154, 166.
Experiments on Plant's. — Robert, Intox., i, 165; Physiologic Methods, Abderhalden,
8, 222; Respiration, Abderhalden, 3, 479; 5, 1271; Gas and Water Movements, ibid., 7, 831;
Biochemistry, ibid., 5, 1263; Sterilization, ibid., 6, 137.
PART II
EXPERIMENTS ON ANIMALS
INTRODUCTORY
Objects of the Course. — The experimental course serves to give a direct,
observational knowledge of pharmacologic actions, sufficient to permit the
student to grasp their essential principles, to obtain a vivid conception
of the effects of the more important drugs, and permit him to follow more
intelligently the more detailed descriptions of the text-books. Incidentally
it also introduces him to the problems of diseased functions (for the effects
of drugs are analogous to these) and to their treatment by therapeutic
agents.
Observations. — The mechanical performance of the experiments, no
matter how carefully they are done, is of relatively small value. At least
equally important are accurate observations and interpretation of the
results. The conditions in animal experiments are much more complicated
than in chemic work. The student must learn to fix his attention on the
main phenomenon, without neglecting anything whatsoever. The more
functions he can embrace in his observations, the more valuable will be the
results and the training. All these observations should be accurately re-
corded. The student must then ask himself the meaning of these results:
What do they really prove? How may they be explained? How could the
several possible explanations be confirmed or refuted? What practical
significance attaches to these effects? In what diseased conditions could
these effects be utilized? How could the toxic effects be treated? etc. The
"questions" may serve as a guide, but the more the student thinks along
these lines, the greater the value of the course.
Note Taking. — The members of a group or subgroup should collaborate
in taking notes, the members alternating as reporters. The notes should
be written out legibly while the exercise is being performed, or immediately
afterward, the different exercises being kept on separate sheets. These
must be handed to the class reporter before leaving the room. The notes
should contain a full record of the observations and discuss the conclusions
which they justify. The technical methods need only be stated in brief
outline, but the doses should always be recorded.
Class Reporters. — Class reporters1 will be appointed for each day, usually
a reporter for each chapter. These are charged with collecting the reports
from the individual groups and with combining these into a comprehensive
report, aiming to present the essential phenomena, and the conclusions which
may be justly drawn from these-, without inning in to extensive details. These
reports will be read and discussed at the weekly conferences of the class, and
tin nott^ taken on these conferences will serve in place of individual notes.
Questions.— The c|ue>tions appended to the chapters must be ansv.
by each student individually, in the standard note-book, within a week after
the class report ha> been read.
i A list of assignments b given in the Appendix.
132 A LABORATORY GUIDE IN PHARMACOLOGY
Demonstrations, Assignments, and Individual Group Work. — It is
desirable that as many of the experiments as possible be performed by the
students themselves. The acquirement of the experimental technic is a
distinct, although perhaps an incidental, benefit. More important is the
fact that many phenomena can be better observed, better grasped, and
better understood when they are produced by the student, studied at leisure,
and varied at his pleasure, than when observed ready-made, at a distance,
and usually only seen for a limited time. Experiments are the more useful
and impressive the more they reproduce the method of solving problems
1 >y actual investigation.
It should, therefore, be aimed to have each group of students perform for
themselves sufficient experiments to illustrate the important principles of
pharmacology and the main actions of the most important drugs. However,
the experiments performed by different groups may profitably be varied
somewhat, so as to illustrate different methods of studying the same phe-
nomenon, and so as to compare and contrast the effects of different drugs.
The experiments performed by the student himself will enable him to under-
stand and evaluate these variations when the results are demonstrated to
him, or when they are reported in the conferences.
Formal demonstrations, however, are also valuable additions to the
individual work. They may be advantageous by presenting experiments
which require special apparatus or which are too difficult for the student;
they may often save time, and, what is very important, they require a
smalter number of animals.
Laboratory Groups. — Partly to save time and animals, and partly to
facilitate the most thorough study of the complex phenomena, the stud'ents
are combined into subgroups (A and B) of three or four men each for the
simpler experiments (frogs, intact mammals, etc.), and into full groups of
six to eight men for the more complex operative experiments. The mem-
bers of the groups should alternate in operating, note taking, etc.
CHAPTER XXXII
LOCALIZATION OF ACTIONS; STIMULANTS AND DEPRESSANTS
Explanatory; Stimulation and Depression.' — Pharmacologic agents act
by increasing or diminishing the normal functions of the tissues. They
never create new functions. Exceptions to this rule are few and, indeed,
only apparent. They depend on the exaggeration of a function which is
normally so slight as to be imperceptible, or which may be latent on account
of unsuitable conditions.
An increase of function is called stimulation. If it is accompanied by
inflammatory phenomena, it becomes an irritation, and is necessarily harm-
ful to the tissue. A stimulation may be harmless, although it tends to pass
into fatigue or exhaustion.
A diminution of function is termed depression. If the function is entirely
abolished, we speak of paralysis. This permits of recovery if it involves
only one function. If all the functions are paralyzed, we have death.
The majority of drugs and poisons produce stimulation at first or in
smaller doses; and depression in larger doses. The principal differences
are found in the relative degree and duration of the stimulation and de-
CHAP. XXXII
LOCALIZATION OF ACTIONS
133
pression. A fairly large number of drugs, however, produce depression
without preceding stimulation; in a few the stimulation is not followed by
depression. In a very few exceptional cases a depression appears to pre-
cede a stimulation; but it is likely that this is merely apparent; for instance,
it may depend on the involvement of different structures.
The immediate and late effects of the same drug, and the action of small
and large doses are, therefore, often opposed. As a general rule, the large
doses produce at first the effects of small doses, even when they have the
opposite effect later. It is customary to distinguish these successive actions
as primary and secondary (and sometimes tertiary), or, preferably, as early
and late effects.
A critical analysis of the actions of drugs shows them to be very simple
in principle: The great majority produce a primary stimulation and second-
raz.
Fig. 7.— Diagram to illustrate possible points of attack of muscle-nerve poisons. The broken
line indicates the afferent mechanism; the solid line, the efferent mechanism.
ary depression of most of the structures to which they may be applied. The
details, however, present an infinite variety, according to the organs and
fum -i inns which are most affected.
Most drugs have a .\r/<r//;r action in this sense. The detailed study of
these selective actions constitutes the special aim of plhinnacodynamics,
and is of great importance t<> the ph\ >i< ian.
Principles of Localization of Action. It is rarely possible t •> underhand
the actions of a drug by the observation of the symptom- which it pro-
duces. Special experiments are required consisting essentially in the
functinnal isolation of structures which might be involved. The following
principles an- generally applicable:
The structures whii h might he involved are considered in the direction
of a retlex chain (Fig. 7).
134 A LABORATORY GUIDE IN PHARMACOLOGY
In case of stimulation the links of this chain are successively paralyzed:
the site of the stimulation is just central to the point at which paralysis
abolished the action. The paralysis is accomplished by section or by drugs.
In case of paralysis, the links of the chain are successively stimulated:
the site of the paralysis is just central to the point where stimulation is
effective. The stimulation is made electrically or by drugs.
In the actual experiments the structures are not taken in the order
named, but according to convenience of technic. It is customary to start
with the nerve-trunk and then to work centrally or peripherally as the
result may indicate.
TECHNICAL NOTES AND REFERENCES
Frogs. — The common grass (leopard) frog, Rana viridis or pipiens, is usually em-
ployed in America. "Medium" frogs, of a body length of 2 to 3 inches, answer very
well; the larger specimens should be reserved for perfusion experiments. The animals
should be kept in a roomy tank, with cold running water and a dry shelf or some stones.
A larger size is needed for perfusion experiments.
Administration of Drugs to Frogs. — Solutions are usually injected into the anterior
lymph-sac. The method of Edmunds and Cushny is recommended: "Lay the animal
back downward in the palm of the left hand. Hold one of its forelegs firmly between the
thumb and index-finger, and the other foreleg between the middle and ring fingers. Draw
its hindlegs downward and hold them against the palmar surface of the hand by means
of the little finger.
"Having the drug in the glass injecting pipet, which is held in the right hand, force
the animal's mouth open with the point. Pass the pipet into the mouth, avoiding the
tongue, which is attached anteriorly, and direct the point toward the floor of the mouth
which with a little pressure it will pierce, entering the lymph-sac. As it is pushed down
the sac the point can be seen beneath the skin of the abdominal wall. The finger is now
removed, and the drug allowed to flow into the sac or, if necessary, blown in."
When very accurate dosage is desired, an exact pipet, furnished with a hypodermic
needle, may be employed. Ordinarily a pipet graduated by the student with file marks
into i c.c. will suffice. The quantity injected should lie between 0.25 and | c.c.
(Injection into the Lymph-sac of the Thigh is described in Chapter XXXVI, Exercise IV.)
Solutions can also be given by the stomach through a blunt glass tube passed down
the esophagus. Many water-soluble drugs (alkaloidal salts, etc.) are absorbed by the
intact skin, and may be administered by painting them on the surface of the skin, or by
placing the entire animal in a jar containing the solution. Gases can be given by placing
the animal under an inverted tumbler.
Weighing of Frogs. — The animal is placed in a tared pasteboard box.
Minimum Fatal Dose (M. F. D.). — This is the dose of a drug which is just sufficient to
kill a unit weight of an average animal-j-often in a given time. It is determined by in-
jecting varying doses into a series of weighed animals. Results which differ widely from
the average are excluded. The author prefers to take the average between the smallest
dose that is fatal and the largest dose that is not fatal. In any case, animals that depart
widely from the average, or that show unabsorbed solution, are excluded.
A more accurate relation exists between the dose and the body surface (Dreyer and
Walker, 1914, Proc. Roy. Soc., 87 B, 319); but the weight relation suffices for all ordinary
purposes. A surface-area formula for man is furnished by Du Bois, 1916, Arch. Int.
Med., 17, 863.
Calculation of Doses. — Doses are usually stated as milligrams of drug per kilograms of
body weight (mg. X kg.). The absolute dose is obtained by multiplying this dose by
the weight of the animal.
Calculation of Dilutions. — i c.c. of a o.i per cent, solution contains i mg. Therefore,
to find the most convenient percentage of .solution, divide the milligrams of absolute dose
by the number of cubic centimeters of solution which you wish to use and multiply the
product by o.i. This gives the percentage. For instance, you wish to inject 90 mg.
in such dilution that from i to 5 c.c. will be needed, i c.c. would require a ^a X o.i =
a 9 per cent, solution; 5 c.c. would require a£ X o.i = a 1.8 per cent, solution. Anything
between these limits will answer. Say that a 5 per cent, solution is at hand. Each cubic
centimeter of this would equal 50 mg. You wish 90 mg., therefore f-{{- =1.8 c.c. With
a little practice one soon comes to judge the proper dilutions without the necessity of
this calculation.
CHAP. XXXII LOCALIZATION OF ACTIONS 135
Work out the following problems and see whether the answers are correct: The dog
weighs 8 kg. You wish to inject 5 to 10 c.c. of each solution. The dose of (a) = o.i gm.
X kg.; (b) = 5 mg. X kg.; (c) = 0.006 mg. X kg. What percentage and how much of
each solution should be used? Answers: (a) 8 c.c. of 10 per cent, or i : 10; (6) 8 c.c. of
0.5 per cent, or i : 200; (c) 4.8 c.c. of o.ooi per cent, or i : 100,000.
Solution Strengths. — The following tabulation will be found convenient:
100. mg. =
10. mg. =
i. mg. =
o.i mg. =
o.oi m£. =
c.c. of
c.c. of
c.c. of
c.c. of
c.c. of
o.ooi mg. = i c.c. of i
10 = 10 per cent.
100 = i per cent,
looo = o.i per cent.
10,000 = o.oi per cent.
100,000 = o.ooi per cent.
1,000,000 = o.oooi percent.
.' Measurement of Solutions. — Quantitative experiments on doses must be made
with chemical accuracy. The graduations of syringes are not sufficiently reliable. The
solutions must, therefore, be measured with pipets, burets, and cylinders. If a syringe
is used, the solution is measured with a pipet into conical glass and drawn from here
into the syringe and injected. The glass is then rinsed with a little water or saline, whu h
is also drawn into syringe and injected. (Rosenau describes the inoculation of precise
quantities, U. S. II\xr. Kali. Hul. No. 19, 1904.)
Behavior of Frogs. — Robert Intox., i, 149.
Motor Stimulation of Frogs, Central and Peripheral. — Ibid., i, 201.
Motor Paralysis of Frogs. — Ibid., 199.
Convulsants. — Ibi<!., 220.
Central Nervous System of Cold-blooded Animals. — Tigerstedt, 24, 153; Frogs, ibid.,
151. 172; successive destruction, ibid., 177.
Turtles. — Ibid., 183; Snakes, ibid., 179.
Removal of brain in frogs and pigeons: Stewart, 961.
Spinal Nerve Roots, Frogs. -Stewart, 957.
Pithing of Frogs.— Frog is held in the left hand and the head bent slightly forward
with the thumb. If the finger-nail is passed lightly along the spine a slight depression
\~01hctory
__
- - Qph c, Lobei
r°me4ulU
Fig. 8. — Diagram of frog's brain.
will be felt back of the head. A narrow-bladed knife is thrust in here, and the brain or
a P. then hi- destroyed by pushing in a still wire. When this is withdrawn the wound
should be stopped with a short pie* e of pointed match to avoid bleeding. A >pe, ial wire
(the thi« kne— ,,f a pen. i! lead and 4 in. hes long) should he reserved for this purpose.
'.rain and medulla alone are destroyed when the animal is to he used for the
observation of reflexes or i in -illation. The cord also when the organ> (heart or n\:
are to be excised.
To destroy ////• hrain nn1\ a line is drawn joining the posterior edee of the tympani.
membranes, and the sl.ull opened in front of this line and the hrain de-.ii. 8).
Decapitation of Frog. — A blade of a stnu rs is pushed into the mouth,
ha< k to the ande of t ! d the skull ( ut away by a -iimle < tit. leaving the lower part.
Anesthesia of Frogs. iv be anestheli/ed under a tumbler by a pl<
i saturated with ether; or. more < onveniently. hy the inje« t ion « ' I per <. ent.
tirethane into the lymph sa« lOehrwall. i. i An h. I'l
Frog-boards.— For dissections or a\ the pit he. 1 ned in
'ion on a . ork hoard. Convenient pi X 4 X T*« i
be Obtained fan applies. These are . ut I- .hes
and D II pine boards of about the same size B mounted
imp.
Preparation of the Sciatic Nerve. The fr md an it
in from the hip to the knee, about the middle of the dot
separating the nuis. 1<^ with th. e nerve is seen as a whitish t ord at the bottom
136 A LABORATORY GUIDE IN PHARMACOLOGY
of the groove. It may be raised by gently passing a thread under it with a frog-needle.1
Care must be taken not to injure it by handling. (Fuller description, Fuehner, 84.)
To ligate the leg exclusive of the sciatic nerve, the nerve is prepared as just described,
and a stout linen ligature is passed below it and tied firmly around the leg, including all
the blood-vi>-i-l.-. The nerve must be protected against drying by covering it with filter-
paper soaked in 0.6 per cent. NaCl. (Details, Fuehner, 86.)
Electric Stimulation. — Inductoria. — Induced currents are generally employed for
stimulation. The Harvard Inductorium is convenient and suffices for most purposes.
The stimulating electrodes are attached to the binding-posts at the end of the metal
rods on which the secondary coil slides. The switch between these rods must be open
when stimulating.
It is convenient to mount the inductorium on a small board, bearing a primary key
connected with the left binding-post. If a dry cell is used, the whole apparatus is con-
veniently arranged in a small box.
The wires from the battery, etc., for the primary current are attached as follows:
For Manising currents, to the two outer binding-posts (or to the key and the right
post).
For single shocks, to the left outer post (or key) and to the middle post.
For single break shocks, connect as for single shocks: (i) Close the secondary key;
(2) open the primary key; (3) open the secondary key; finally (4) open the primary key,
which gives the single break shock. ("Cut-out Keys" are described on p. 793 of the 2d
edition of this book; and by Kingsbury & Dresbach, 1910, Quart. Jour. Exp. Physiol.,
3, ii i; Laidlaw, 1913, Jour. Pharmacol., 4, 461.
The strength of the secondary current is regulated by the distance of the secondary
from the primary coil, and by revolving the secondary on its axis: the greater the distance
Fig. 9. — Diagram of individual switchboard. The wires leading to the apparatus are attached
at aa, bb, or cc.
of the coils, and the greater the angle, the weaker the shocks. (The specific graduation
of inductoria is described by Martin, Amer. Jour. Physiol., 1911, 33, 212; 1915, ibid., 36,
223.)
Electrodes. — The ordinary (Harvard, platinum) electrodes are usually employed.
For deep-seated nerves the shield electrodes (Harvard) are advantageous. For direct
stimulation of muscle fine insulated wires are connected with the secondary posts, the
other end, freed from insulation, being thrust through and hooked around the muscle.
Non-polarizable and brush electrodes (Mottram, 1915, Jour. Physiol., 49, Proc.) are
needed only for special problems.
Source of Current.— Ordinary dry cells may be used; they are conveniently mounted
in a little box under the inductorium. Any other type of cells may be employed. If a
steady current is required, Daniell cells or a storage battery are essential.
Street Current. — The direct current is very convenient, especially for class work. It
is cut down to the required voltage on the Wheatstone-b ridge principle, as described by
D. E. Jackson, Jour. Amer. Med. Assoc., 58, ion, 1912; by v. Hess, 1914, Science, 40,
566; and by Y. Henderson, 1915, ibid., 41, 910.
Any number of coils, etc., may be supplied from a single closed circuit over each table,
the circuit passes through a spiral of iron wire ("stove-pipe"), 13 cm. long, wound on a
\ inch rod. The spiral is mounted on an asbestos board and connected with binding-
posts, as shown on the diagram (Fig. 9).
Flexible wires, attached to each pair of posts, conduct the current to the coils or
other apparatus.
1 Frog-needles are made by heating a stout sewing needle % inch from the blunt end until it
can be bent at right angles and fixing the point in a convenient wooden handle (penholder).
CHAP. XXXII LOCALIZATION OF ACTIONS 137
Perfusion of Frog's Aorta. — Lay the pithed frog on his back, the head toward the
operator. With scissors and forceps cut away a flap of skin, from the jaws to the thighs,
deflecting it downward. Remove the sternum. Cut away a flap of the abdominal wall
and also turn downward. Pin the frog to a board. Tie a small cannula into the peripheral
end of the aorta; fill with saline solutions and connect with perfusion bottle.
Observation of Reflex Time. — The frog (usually with brain and medulla pithed) is
held with forceps or suspended from a hook passed through the lower jaw, and one or both
hind feet immersed in a dish containing 5 per cent, acetic acid or j per cent. HC1.
The reflex time is the time elapsing between the immersion and the withdrawal of the
foot. The average of several observations should be taken, the acid being washed off after
earh test, and a short interval of rest must be given. (Further discussion, Robert Intox.,
i, 191.)
Experiments on Motor Nerves. — Kobert Intox., i, 169; Stewart, 780.
Muscle-nerve Preparation. — The frog is pithed through brain and cord. It is then
held up by the legs so that the anterior part of the body falls down. The scissors are
thrust through the body a little anterior to the angle and the whole body is cut off. By
grasping the skin with a cloth it can be readily removed from the legs. The two U :
then cut apart just in the median line. The iliac bones (the two bones at the sides) are
cut away. Each portion is then turned with the posterior surface upward, and the
muscles of the thigh are pulled apart with the fingers. The sciatic nerve will be seen lying
at the bottom of the groove. It is carefully dissected out with a few cuts of the scissors,
from the spinal canal to which it is attached, to the knee. The thigh is then cut off so as
to leave a short piece of the femur attached to the knee.f — A blade of the scissors is then
thrust under the tendo Achillis, and pushed as far as possible toward the toes. The
tendon is then cut off at this point. The tibial bone is also divided close to the knee. —
In this way a preparation is formed consisting of a small piece of bone of the spinal column
attached to the sciatic nerve, a bit of the femur, the gastrocnemius muscle, and the tendo
Achillis. These preparations must be carefully kept from drying by wrapping in filter-
paper soaked in normal saline solution.
If the drugs are not to be applied directly to the muscle, the skin may be left on the
preparation. If the poison is to be applied only to the nerve, the operation need only to
be carried to f.
Gastrocnemius Preparations. — If the muscle alone is to be observed, the prepara-
tion of the nerve may be dispensed with. The leg is amputated just above the knee. If
the muscle is not to be exposed to the poisons, this preparation may be used as it is.
Otherwise the skin may be removed and the muscle prepared as in — to — of the last
paragraph.
sometimes desirable to obtain a record of muscular contractions while the cir-
culation through the muscle is intact. For this purpose the pithed frog is pinned on the
board, dorsal surface up, and a ligature is passed through the tendo Achillis and attached
to the lever.
Protection Against Drying. — The muscle and nerve must be carefully protected
from desiccation. This is superfluous if the preparation is covered by skin; otherwise, it
may be wrapped in filter-paper saturated with normal saline solution. The nerve may be
painted with the solution, using a < amel's hair brush or swab. If it is necessary to keep
the moisture constant, the preparation is covered by a tumbler or bell-jar lined with moist
filter-paper. A "moi-t ( number" i- made by the Harvard Apparatus Company.
Direct Application of Drugs to the Muscle or Nerve.— This may be done, according
to < ir. urn-tames, by dipping the part into the solution, or by painting will-,
hair brush, or by allowing the solution to flow over the part from a pipet. The penetra-
tion of solution- into the muscle may be facilitated by scarifying the sheath.
Gases may be applied by plat ing the preparation. <>r any part of it, into a tube through
whit h the mni-t ga> is flowing 'Harvard gas chamber).
Technical Reference. Must le-nerve preparation, Fuchner, 120.
Normal Saline Solution for Frogs.— This is a 0.75 per cent, solution of sodium ( hlorid.
EXERCISE I.— LOCATION AND TYPE OF CONVULSIONS (FROGS)
PORTER I, A)
Explanatory. — Tlr e involves the application of the principles
•I. Both strychnin and pieroto\in cause convulsions. The
action mi^ht conceivably U- located in the in the luain.
medulla, spinal . ior endings, or muscle-fibers. If it is central,
138 A LABORATORY GUIDE IN PHARMACOLOGY
it could be due to direct stimulation or to increased sensibility to reflex
impulses.
The student will determine the correct explanation by his experiments.
The type of the convulsions, when once seen, gives a very plain hint of
the probable location of the action. The student should tabulate the dis-
tinctive differences between strychnin and picrotoxin. This will be facili-
tated by the use of the following terms:
Opisthotonus: Body arched backward.
Emprosthotonus: Body arched forward.
Clonic Convulsions: Intermittent, jerky.
Tonic or Tetanic Convulsions: Permanent stiffening.
Quite a number of poisons produce the same effects as strychnin; for
instance, small doses of caff ein ; morphin also produces the same action after
a time.
Large doses of caffein and veratrin produce effects which resemble those
of strychnin superficially. The action of caffein, however, is due to rigor,
for it persists after cutting the nerve, and the muscles are inexcitable. Vera-
trin acts directly on the muscle-fiber, for even the isolated muscle remains
contracted for a long time whenever it is stimulated.
Experiment i. (Demonstration) Strychnin Convulsions. — Inject into
the lymph-sac of a frog (Tech. Notes) J c.c. of TV per cent, strychnin. Ob-
serve the type of the convulsions carefully (illustrated in Fuehner, 72).
Note when they appear; that the legs are extended and the arms flexed; the
frog may be held horizontally by the feet. The convulsions intermit, the
frog being paralytic between the spasms. The spasms may start with a
cry.
The convulsions are typical of spinal stimulation (increased reflex
excitability of the spinal cord).
Question.— Describe the effects of strychnin and draw a sketch of the
frog in the typical tetanus.
Experiment 2. (Optional) Bio-assay of Strychnin. — Frogs are a more sensitive test
for strychnin than are the chemic reactions (Ranke, 1879, Arch. Path. Anal., 75), espe-
cially if the solutions are somewhat impure. For the American leopard frog, with injection
into the lymph-sac, the tetanic dose of strychnin sulphate is about o.i to 0.15 mg. per
100 gm.; the M. F. D. (Tech. Notes) is 0.555 mg- P6*" 100 gm. Decereberated frogs are
more sensitive.
Young White Mice give a still more delicate test, those of 4 to 4.5 gm. responding to
the hypodermic injection of 0.002 mg. by tetanus within ten minutes; 0.005 mg- being
fatal. Tremor of the tail is especially characteristic.
Experiment 3. (Demonstration) Nature of the Stimulus. — Note, on the
above frog, that the convulsions appear, as a rule, only when the animal
is stimulated.
Note that the following stimuli are effective — touching, jarring the table,
sound (clapping hands), electric stimulation of the skin.
Lower a foot of the frog into 5 per cent, acetic acid : the leg is drawn
up as in a normal reflex, but there are no convulsions.
Float the frog in a bath of oil: the convulsions are allayed.
Questions. — (a) Does the strychnin stimulate the convulsion centers
directly?
(b) Are all varieties of stimuli effective?
Experiment 4. (Optional) Tetanus Threshold. — Pith the brain of a frog (Tech.
Notes). Remove the skin from the muscles of one leg. Expose the sciatic nerve (Tech.
Notes). Expose the intestines. Set up an induction coil (Tech. Note). Determine the
(HAP. XXXII LOCALIZATION OF ACTIONS 139
weakest current which produces reflex contraction of the sound leg when applied to the
skin of the foreleg and to the exposed muscles, nerve, and intestine. In stimulating the
intestines, guard against escape of current to the sciatic plexus.
Strychninize the frog, and when convulsive, test the threshold for tetanus in these
various situations. The strongest current will be required for the intestines, the weakest
for the nerves of the skin.
Question. — Why are the intestines and muscles less liable to produce
convulsions?
Experiment 5. (All A Groups) Location of Strychnin Tetanus. — Inject
.1 . of TV per cent, strychnin into a frog.
(a) Immediately after the convulsions appear, destroy the brain (Tech.
\< 'tes) : the convulsions continue.
(b) Destroy the medulla in the same frog: the convulsions continue.
(c) Cut all the muscles of one leg through to the femur: this leg ceases
to participate in the convulsions.
(d) Destroy the spinal cord: the convulsions cease.
(e) (Optional) Ligate the leg of another frpg, exclusive of the sciatic nerve. Inject
i c.c. of ^ per cent, strychnin below the ligature: no convulsions.
Formulate conclusions justified by each of these experiments as to the
site of the strychnin action.
Questions. — (a) Is the strychnin action situated in the brain? (b) In
the medulla? (c) In the muscles? (d) In the afferent nerve-endings? (e)
Where is its situation?
Experiment 6. (Optional) Location of the Strychnin Action Within the Spinal Cord. —
Strychninize the cervical spinal cord, without letting the poison reach the lower portions
of the cord:
Insert the lower blade of the scissors in the mouth of a frpg, and cut away the i-ntire
top of the head, as far back as possible, from the angle of the jaws.
Stop the circulation by opening the frog and excising the heart.
Apply a pledget of cotton soaked in o.i per cent, strychnin to the exposed cervical
section of the cord.
Test the reflexes by pinching the fore- and hindlegs.
he experiment is successful, the following results will be obtained:
(a) Pinching the hindlegs causes a normal n-llrx.
(b) Pim-hing the forelegs produces convulsions of the entire animal.
Experiment 7. (Demonstration) Inhibitory Influence of Cerebral
Lobes on Spinal Convulsions (Acid Fuchsin).— Weigh two frogs (Tech.
Note). Use the one (A) as control. From the second (B) remove the an-
terior half of the brain by cutting with scissors from the angle of the jaw-,
and just back of the eyes.
Inject into the lymph-sac of each frog arid fuch sin, 0.03 c.c. of 5 percent,
per gram. B will -how strychnin-like convulsions within lift mi min
in A thc-e \\ill be delayed for one to twenty hours (the further observation-
may be assigned to one of the A groups). (Barbour and Abel, iqio, Jour.
I'harmacnl.. .-. if.;; the action i- al-»> much quicker and occur- with much
ler do-rs. if the heart has been excised ; Joseph and Meltzer, 1911, Jour
I'harmai «>].. J, i E
Questions. — (a) Why does the removal of the brain hasten the onset of
the convulsions? (/» What effect has the brain on the reflexes of normal
animals?
Experiment 8. (Demonstration) Picrotoxin (Medullary Convulsioi
Inject into the lyniph-.-ac of the frog 1.5 c.c. of I : 250 solution of picrotoxin
140 A LABORATORY GUIDE IN PHARMACOLOGY
(a) Convulsions occur only after a period of depression lasting to half an
hour. The animal goes through a regular cycle of motions. (Illustrated in
Fuehner, 73.) A characteristic feature is that the legs are abducted in one
stage. The animal may turn a somersault. The abdomen may be bloated
with air. Between the spasms the animal is depressed.
(b) The convulsions may occur in the absence of stimulation.
(c) Destroy the brain: the convulsions persist.
(d) Destroy the medulla: the convulsions disappear.
(The M. F. D. for medium frogs is about 0.5 mg.)
Questions.— (a) What are the characteristic differences between the
strychnin and picrotoxin movements?
(b) Are the picrotoxin convulsions due to direct stimulation, to increased
reflex excitability, or both?
(c) Where is the picrotoxin action located?
Experiment 9. (Optional) Other Central Convulsants. — The following may be used
on frogs (lymph-sacs):
For Spinal Convulsions. — Hydrastin, i to 2 c.c. of i : 1000.
For Medullary Convulsions. — Ammonium carbonate, 2.5 c.c. of i per cent.; camphor,
i c.c. of 10 per cent.; phenol, i c.c. of i per cent.
Experiment 10. (Demonstration) Veratrin (Muscular Spasm). — Inject
into a frog 0.5 c.c. of i : 10,000 veratrin (= 0.05 mg.). When the effect
is fully developed the animal sits normally, but when it jumps the leg
remains extended as in tetanus. This relaxes very slowly. If the animal
is made to jump repeatedly, its behavior becomes more and more normal,
but if it is allowed to rest the stiffness returns.
Pith the brain: the condition remains unchanged.
Divide the tissues of one leg to the bone, and stimulate the muscle with a
single shock (Tech. Notes) : the muscle still shows prolonged stiffening.
Questions. — (a) How could you distinguish between a strychnin tetanus
and a veratrin contracture?
(b) Is the veratrin action central or peripheral? Why?
Experiment u. (Demonstration) Caffein Convulsions. — Inject a frog
with 10 mg. of caffein (i c.c. of i per cent.) : marked stiffness and sometimes
strychnin-like tetanus.
Experiment 12. (Demonstration) Caffein Rigor. — Pith a frog, and in-
sert a cannula into descending aorta (Tech. Notes). Inject | to i c.c. of i
per cent, caffein: immediate rigor. The muscles appear white and hard,
do not respond to electric stimulation, and are acid to litmus.
Question. — How could one show that the caffein stiffening is not due to
central tetanus?
EXERCISE II.— CENTRAL DEPRESSANTS ON FROGS
(REPORTER II, D)
These produce, successively, quietness; inco-ordination, so that the frog
cannot readily turn from its back; muscular relaxation; anesthesia; absence
of reflexes.
. The paralysis can be shown to be central by stimulating the sciatic nerve:
this should evoke a normal contraction of the leg.
Experiment i. (Group I-B) Morphin (Descending Central Paralysis).—
Inject into the lymph-sac of a frog 50 mg. of morphin sulphate (ij c.c. of 4
per cent.) . Observe the symptoms (which correspond to the ablation of the
CHAP. XXXII LOCALIZATION OF ACTIONS 141
central nervous system at successively descending levels). The animal
at first becomes quiet, and does not move spontaneously; it sits erect, how-
ever, and jumps if stimulated. Place the animal on a small board, and tilt
the head-end slowly up: the animal will climb up the board (if observed suffi-
ciently early; later it will not do so). Laid on its back it recovers its normal
position. Place the morphin frog and a normal frog in a tumbler filled with
water, and invert this over a large jar filled with water (not admitting any
air into the tumbler). Both frogs will rise to the top to breathe; but the
normal frog, finding no air, will dive down and out of the tumbler; the
morphinized frog remains. Remove it from the tumbler and observe
that it can swim. Remove from the water. As the action of the poison
progresses, the frog will sit more flat. Laid on its back, it makes ineffectual
efforts to turn. Still later the frog lies quite flat , makes no effort to turn, and
cannot swim. On pinching the toe, the leg still contracts. This shows that
the cord and the peripheral sensory and motor nerves are not paralyzed.
Lay the frog aside; in the course of some hours or on the next day the
animal is found in typical strychnin convulsions. (One of the frogs may re-
ceive the morphin several hours beforehand, and the convulsions demon-
strated.)
Question. — In what order are the nerve-centers depressed by morphin?
Experiment 2. (Optional) Decerebration on Morphin-tetanus. — Decapitate a frog
(Tech. Note). Inject 10 mg. of morphin (i c.c. of i per cent.). Tetanus occurs hi one-
half to six hours, while a normal frog would require about twenty-four hours (compare
with Exercise I, Experiment 7). Cold also favors the onset of the convulsions (Githens,
1912, Proc. Soc. Exp. Biol. Med., 10, 40).
Experiment 3. (Group II-B) Alcohol. — Inject into the lymph-sac of a
2 c.c. of 25 per cent, alcohol: paralysis; abolition of reflexes; depressed
respiration. Stimulate sciatic nerve (Tech. Note) : normal response.
Questions. — (a) Describe the cause of the depression.
(b) Is it central or peripheral?
Experiment 4. (Group III-B) Chloral. — Inject into the lymph-sac of
the frog i c.c. of 2 per cent, chloral. Observe as for alcohol (Experiment 3).
Experiment 5. (Group IV-B) Ether. — Place a frog under an inverted
tumbler containing some cotton saturated with ether: effects as with alco-
hol (Kxperiment 3).
Experiment 6. (Group V-B) Magnesium. — Inject into lymph-sac 0.8
c.c. of 25 per cent. MgSO4 for each 10 gm. of frog. Complete anesthesia in
an hour. Recovery by next day.
Other Central Depressants which could be substituted for the above are:
Chloroform, i c.c. of 20 per < ent., in olive oil.
Scopolutnin, i e.e. of i per * cnt.%
Codrin, i e.e. of i per cent.
//';;, i C*C* of i |>rr.ciit. This opium alkaloid arts like stryihnin.
Experiment 7. (Optional) Comparative Narcotic Activity. — This is best determined
pn small fish or t.ulpolc- p|.i< r<l in s,,luli»ii- <>i dilTerenl < on. nitration. The 'Ynd point"
is complete abolition of all movements r\< ept respiration, and recovery in i
water. Three animals in about 200 c.c. of solution arc tiM-d for rai h lails.
Fuehncr, 52.)
Comparative Analgr can be determined in man as described by Macht,
Herman, and Levy, 1916, Jour ! her., 8, i.
Experiment 8. (Demonstration) Anesthesia of "Salted Frog."— Re-
move the blood from I he vessels of a frog by perfusion of the aorta with
142 A LABORATORY GUIDE IN PHARMACOLOGY
oxygenated saline solution: the frog acts as if normal. Expose to ether as
in Experiment 5 : anesthesia occurs just as in a normal frog.
Question. — Can the action of the anesthetic be attributed to changes
in the blood or circulation?
EXERCISE III.— REFLEX TIME
(REPORTER III, A)
This is determined on decapitated (why?) frogs by Tuerck's method
(Tech. Notes) by immersing the foot in 0.5 per cent. HC1, and noting the
time until it is retracted. The acid is washed off and several determinations
are made.
Experiment i. (Group i-A) Demulcents. — Determine the reflex time
of normal decapitated frog, comparing 0.5 per cent. HC1, and 0.5 per cent.
HC1 containing 15 per cent, of acacia. The reaction is greatly delayed.
Question. — Why does acacia delay the reaction?
Experiment 2. (Group II-A) Alcohol. — Determine the normal reflex
time of decapitated frog. Inject into lymph-sac 50 mg. of alcohol (0.5 c.c.
of 10 per cent.) and again determine reflex time at intervals.
Question. — What is the effect of alcohol on reflex time?
Experiment 3. (Group III-A) Urethane. — Proceed as in Experiment 2,
injecting urethane, 0.2 gm. in 2 c.c.
Experiment 4. (Group IV-A) Morphin. — Proceed as in Experiment 2,
injecting 10 mg. of morphin (J c.c. of 4 per cent.).
Experiment 5. (Group V-A) Strychnin. — Proceed as in Experiment 2,
injecting 0.02 mg. of strychnin (0.2 c.c. of i : 10,000).
(Optional) Other Drugs which May Be Tested on Reflexes:
Caffein, % c.c. of i per cent.
Potassium Chlorid, 0.3 c.c. of 5 per cent.
EXERCISE IV.— DEPRESSION OF MOTOR ENDINGS (CURARE ACTION)
(REPORTER IV, D)
Explanatory. — To determine whether a motor paralysis is central or
peripheral the sciatic nerve is exposed and stimulated electrically. If
there is no response, the paralysis is peripheral. If the muscle contracts, the
central seat of the paralysis is located by successive stimulation of the
cord and medulla.
A peripheral paralysis may be in the nerve-trunk, the endings, or the
muscle-fiber. No drug is known which acts selectively on the motor nerve
trunk when applied systemically. The possibility of this action may be
excluded by the curare experiments described below. If the motor endings
are paralyzed, the muscle will contract if the electrodes are laid directly upon
it. This effect is produced most typically by curare; but it is also shared to
a minor degree by strychnin, morphin derivatives, coniin, lobelin, camphor,
organic ammoniums, magnesium, etc. These drugs, however, have other
actions which are much more powerful, and which generally kill the animal
in doses much smaller than those required to produce the curare effect.
This may, therefore, be very incomplete, or may be demonstrable only by
local application to frog's muscles.
Technical References — Tigerstedt, i.i, 36; 2.4,323; Preparation of Curarin, Abder-
halden, 2, 942; on small scale, Boehm, 1910, Arch. ges. Physiol., 136, 203; Curare Paper
(for small doses), Jacobj, 1907, Deut. med. Woch., i, 1540.
CHAP. XXXII LOCALIZATION OF ACTIONS 143
Experiment i. (Demonstration) Symptoms of Curare Poisoning. —
Inject \ to i c.c. of a \ per cent, solution of curare into the lymph-sac of a
frog, repeating the dose even- twenty minutes if necessary. Note the general
symptoms: the reflexes disappear and the frog shows a general muscular
paralysis, but without the preceding cerebral depressions which were ob-
served with morphin. (With some samples of curare, strychnin tetanus
precedes the paralysis.)
Experiment 2. (Demonstration) Seat of Curare Action. — When the
reflexes have entirely disappeared in the above frog, isolate and stimulate
a sciatic nerve. There is no response (or if the poisoning is incomplete,
only a slight contraction) . The paralysis is, therefore, peripheral to the cord.
Apply the electrodes directly to the muscle: there is a strong, normal con-
traction.
Questions. — (a) Is the curare paralysis central or peripheral? Wrhy?
(b) Does it act on the muscle-fibers? Why?
(c) Where must its action be located? Why?
Experiment 3. (Demonstration) Claude Bernard Experiment. — Take
another frog, pith its brain, and ligate one leg, excluding the sciatic nerve.
Inject the dose of curare used in (i) into the lymph-sac, and allow it to
develop its action. Stimulate the sciatic nerve of both legs: the unligated
leg does not respond; the ligated leg contracts. Direct stimulation of the
muscle produces contraction in either leg. The ligature which prevented
the action of the curare excluded the poison from the nerve-endings, but
not from the greater part of the nerve-trunk.
Questions. — Does the curare act on the motor trunk? Why?
Experiment 4. Curare Action on Muscle-nerve Preparation. — The con-
clusions of Experiments 2 and 3 may be arrived at more simply, and on one
animal, as follows: Fit a slide across a small evaporating dish containing the
drug dissolved in normal saline; the solution should not reach the slide.
Make two muscle-nerve preparations (Tech. Notes) from a fresh frog; deter-
mine the threshold current. (Tech. Notes) which will give contraction when
applied to the nerve and directly to the muscle. Lay the muscle of one
preparation on the slide, letting the nerve dip in the solution. Lay the nerve
of the otlu-r preparation on the slide, letting the muscle lay in the solution.
Remove the preparations every five minutes, testing their excitability as
described above; replace them, and repeat as often as necessary. Present
the results in tabular form :
Stimulation of: Nerve in solution. Muscle in solution.
Dislanfe of coils: Muscle. Nerve. Muscle.
• laying in >oluti<m
Five minutes.
Ten minute-
It' the solution contains a. drug with curare action, the nerve which has
lain in the solution retain- it- r\i itahility. The preparation ot" which the
muscle has lain in the solution becomes inexc itable to stimulation by tin-
nerve; the muscle itself retains it- exi it ability.
The following solutions may he u>nl. employing a muscle-nerve prepara-
tion from tin frogs used in a previous exix-riment :
(Group I-B): Curare, i : looo in N. S. ")
(Group I I-B): Nicotin. i : 1000 in N. S. /• Use for Experiment 5.
(Group III-B): Magnesium Sulphate, S P61" cent ^
144 A LABORATORY GUIDE IN PHARMACOLOGY
Questions. — (a) How does this show that the drug paralyzes the motor
endings?
(b) Is the action an actual paralysis or a "block"?
Experiment 5. (Groups of Experiment 4) Antagonistic Action of Phy-
sostigmin. — Lay the muscle which has been depressed by curare, etc., in
physostigmin, i : 1000 N. S. Test excitability from time to time: some
recovery occurs.
Question. — Does the physostigmin act on the drug, or on the functions?
Experiment 6. (Optional) Antagonism of Physostigmin and Curare in Rabbits. —
Anesthetize a rabbit with Paraldehyd, i gm. per kilogram, by stomach-tube. Prepare
for artificial respiration. Connect the jugular vein with an injection buret.
Inject into the vein physostigmin, 5 mg. per kg. (5 c.c. per kg. of i : 1000). This
produces fibrillary twitchings.
Divide the sciatic on one side: the twitchings persist. Inject curare, f c.c. per kg.
of J per cent.: the twitchings disappear.
Gradually increase the curare until the respiration stops (being ready for artificial
respiration). Note that sciatic stimulation is again ineffective.
Inject physostigmin (several doses if necessary) : excitability reappears.
Fig. 10. — Nicotin. Successive positions of frog poisoned with nicotin.1
Questions. — (a) Is the action of the physostigmin central or peripheral?
(b) How could one treat curare poisoning?
Experiment 7. Direct Paralysis of Muscle. — Use the arrangement of
Experiment 4, but employ the following:
(Group IV-B): Saponin, i : 1000 N. S.
(Group V-B): Apomorphin, i : ioooN. S.
Question. — What is the site of the depression in these cases?
(Other protoplasmic poisons also paralyze the muscle-cells directly; e. g.,
cocain or quinin (i : 1000 to i : 100 solutions). Apomorphin and copper
salts have the same effect, even when injected systemically.)
Experiment 8. (Demonstration) Systemic Administration of Nicotin. —
(a) Inject into the lymph-sac of a frog i mg. (= i c.c. of o.i per cent.)
nicotin. Note that the frog becomes gradually depressed, assuming the
characteristic positions illustrated in Fig. 10. (Note the peculiar twitching
of the muscles. Divide one sciatic nerve: the twitchings cease. Stimulate
1 Further illustrations in Fuehner, 75.
CHAP. XXXII LOCALIZATION OF ACTIONS 145
the nerve: they reappear. The seat of this action is, therefore, in the muscle
or endings, but it can only find expression if the nerve is stimulated from
the brain or electrically.)
(b) Make a muscle-nerve preparation from the frog, and test the quan-
tity of current required i /. e.t the distance of the coils) to obtain a contrac-
tion if the electrodes are applied to the nerve, and if they are placed direct ly
on the muscle; less current is needed on the muscle. Since the reverse is tin-
case in a normal preparation, it is evident that the nicotin must have de-
pressed the nerve-trunk or the endings.
The position of the frog (folding of hindlegs over back) is very charac-
teristic of nicotin, and serves to distinguish it from all related poisons;
-jV mg- may be demonstrated in this way.
Experiment 9. (Demonstration) Nicotin in Tobacco Smoke. — Take a
small tubulated bell-jar fitted with a doubly perforated stopper. One of the
perforations bears a tube reaching just below the stopper. Into the other
opening of the cork fit a thistle-tube, which should reach to near the bottom
of the jar. Fill the thistle with tobacco. Place a frog under the bell-jar;
fix the latter with vaselin on a glass plate. Light the tobacco, and aspirate
the smoke into the jar. The frog will show the same symptoms as in nicotin
poisoning, since the nicotin is the main active ingredient of tobacco smoke.
Experiment 10. (Optional) Demonstration of Curare Action in Other Drugs. — The
following may be used similarly to nicotin:
Locally
In lymph-sac. (in 0.75 saline).
Camphor o.i gm. Saturated
Lobelia 2 " 4 per cent.
Conium 2 4
Coniin 10 mg. 0.2 "
Lobelin 10 " 0.2 "
Magnesium Sulphate 1.5 c.c. of 50 per cent, solution.
Strychnin i per cent.
EXERCISE V.— PERIPHERAL SENSORY PARALYSIS (LOCAL ANESTHESIA)
(REPORTER V, D)
Explanatory. — Sensory paralysis is evidenced by failure to respond to
sensory stimuli (motor paralysis having been excluded by stimulation of tin-
sciatic nerve). Central paralysis is excluded by stimulation of an afferent
nerve-trunk. If this proves effective, the sensory paralysis is peripheral.
Thi> may involve the nerve-fibers, endings, or sensory end cells. It is not
always possible to distinguish absolutely between these. Nerve-trunks are
only paraly/.ed by direct application. As a general rule, this paralyzes
both sensory and motor fibers, but the sensory fibers are affected much more
readily. It is somewhat easier, however, to (Union- trate the paralysis of the
motor functions, as in the experiment- lielow.
Sensory depressant - are ut ili/ed for local anesthesia. General anesthesia
may be produced by injecting them into the subdural (anal. It must be
remembered that they m-ed to be brought into direct contact with the
-tructure to be paralyzed. They are <|iiite inactive on surface* from which
they are not absorbed, such as the intac t mammalian >kin. On the other
hand, they are effective on mucous mrmbrano and the frog's skin. In
other sjtuati.'i)> they are used by hypodermic injection or painted on the
nerve, or injected under its sheath. Cocain and its substitutes are the l»e>t
nplcs of local anesthetics.
146 A LABORATORY GUIDE IN PHARMACOLOGY
None of the peripheral sensory depressants are sufficiently selective to
act from the circulation without producing general intoxication. They are
therefore used locally, and in the case of local anesthetics the action is
further confined to the place of application by restricting the circulation
with a bandage or by suprarenal alkaloid.
Sensory anesthesia may also be produced by very powerful sensory
stimulation. Most irritants are succeeded by anesthesia. Aconite and
menthol are examples.
Technical References. — Sensory Paralysis of Frog, Robert, Intox., i, 223; Eye, ibid., i,
215; Tripolar and other blocks, Gruber, 1913, Kansas Univ. Sci. Bui. 17, Nos. 10 and u;
Amer. Jour. Physiol., 31, 413.
Strength of Local Anesthetics. — The strength of local anesthetics may
be tested and compared by the following method:
(a) Rabbit or human cornea (Experiment 2) ; (b) acid-reflex, frog (Ex-
periment 4) ; (c) conductivity of the sciatic motor nerve, frog (Experiment
5); (d) conductivity, motor and sensory (reflex), of rabbit's sciatic; (e)
infiltration (Experiment 12).
The methods (a) and (b) estimate the anesthetic power for mucous
membranes, where absorption is a factor. They give concordant results
(Fromherz, Arch. exp. Path. Pharm., 76, 257). The other methods, es-
pecially the last, estimate the anesthetic power independent of absorption.
(The various methods are described by Fuehner, 165.)
Experiment i. (Demonstration) Anesthesia of Cornea. — Touch the
cornea of a rabbit (or other animal) with a stiff bristle (mounted at right
angles on a wooden rod, Fuehner, 168) , and note the winking reflex. Apply in
one eye a drop of i per cent, cocain, in the other a drop of i per cent, quinin-
urea hydrochlorid. Note that reflex is gradually abolished. This method
also shows, by the behavior of the animal, whether the drug is irritant.
Questions. — (a) Which of the two drugs is the more powerful anesthetic?
(b) Does either produce irritation?
Experiment 2. (Optional) Exact Estimation of Anesthetic Power on Cornea. —
This involves the use of a series of straight hairs of different diameters, i or 2 inches long,
cemented on the end of small wooden sticks (v. Frey's "Reizhaarmethode").
The sensitiveness of the corneas is tested toward a series of, say, five such hairs, the
pain-reaction of the human cornea being the most delicate reaction. A drop of the
solution (cocain, i per cent.; novocain, 2 per cent., etc.) is then placed in the eye, and the
tests with the hair repeated at intervals, observing the time when anesthesia appears
and disappears. Both corneas may, of course, be used for different solutions.
The force exerted by the different hairs may be measured by pressing them against
a balance and counterbalancing with weights. This weight is divided by the square area
of the cross-section of the hair, calculated from its micrometer measurement.
Experiment 3. (All Groups) Anesthetic Action on the Tongue. — This
serves as the rough qualitative test.
Place the drug on the tip of tongue (or saturate a small piece of filter-
paper with the solution and place on tongue) and test the sensibility to touch.
The (A) groups may use a drop of i per cent, cocain; the (B) groups a
drop of Tr. Aconite. In the latter the anesthesia is preceded by prickling.
Question. — Would aconite be suitable as an anesthetic for eye-work?
Experiment 4. (B Groups) Anesthetic Action on Frog's Foot. — Test
the reflex time (Tech. Note) of a decapitated frog (0.5 per cent. HC1). Dip
one foot in i per cent, cocain, the other in i per cent, solution of some other
anesthetic (see below).
CHAP. XXXII LOCALIZATION OF ACTIONS 147
Again test the reflex time at intervals of five minutes.
Group I-B — Test i per cent. Novocain.
Group II-B — Test i per cent. Stovain.
Group 1 1 I-B — Test i per cent. Quinin-urea Hydrochloric!.
Group IV-B — Test i per cent. Tropacocain.
Group V-B — Test i per cent. Alypin.
Question. — Arrange the drugs in the order of anesthetic efficiency for
mucous membranes (the frog's skin is virtually a mucous membrane).
Experiment 5. (A Groups) Paralysis of Nerve-fibers on Direct Applica-
tion.— Make two muscle-nerve preparations with long nerves (Tech. Notes)
from a frog (one used in an earlier experiment). Determine the threshold
stimulus of the sciatic. Paint a short stretch of one nerve with i per cent,
cocain; paint the other with another anesthetic.
Again determine the threshold stimulus in five-minute intervals. When
anesthesia is complete, wash the nerve with normal saline and note that its
excitability gradually returns. The method really measures the depression
of the motor-fibers, which are more resistant than the sensory fibers. How-
ever, since the two are generally parallel, it is an admissible, though indirect,
measure of anesthetic power. For the comparison the following solutions
are to be used:
Group I-A — 2 per cent. Hydrocyanic Acid.
Group II-A — i per cent. Stovain.
Group III-A — i per cent. Quinin-urea Hydrochlorid.
Group IV- A — 25 per cent. Magnesium Sulphate.
Group V-A — Perform Experiment 6.
(Optional) o.i per cent. Chloroform.
lion. — Arrange the drugs in order of anesthetic efficiency for neural
application.
Experiment 6. (Group V-A) Synergism of Epinephrin and Cocain.—
Proceed as in Experiment 5, using four muscle-nerve preparations, as fol-
lows (c and d should be from the same frog) :
Lay Nerve (a) in 2 per cent. Morphin in N. S.
Lay Nerve (b) in i : 1000 Epinephrin in N. S.
Lay Nerve (c) in i : 100 Cocain in N. S.
Lay Nerve (d) in i : 1000 Epinephrin containing i per cent. Cocain in N -
iMrrminu the threshold of stimulation in five-minute intervals.
Questions. — (a) Does epinephrin hasten or increase the efficien
cocain?
(b) Is epinephrin an anesthetic?
(c) How would you explain the synergism?
(d) Is morphin a local anesthetic?
(Optional) Experiments on Oilier Mixed Local A ntstkctics.— See Zorn, 1913, Zs. exp.
Path., 12, 529.
Experiment 7. (Optional) Anesthesia by Gases. — Draw tin- i -MII-. !«•
preparation through a «as < h.imher. ami expose it to the vapors of ethi .ihon
mulation l>e< on
Experiment 8. (Optional) Depression of Conductivity by Ether.— The la. t th.
.n.lu. tivity M well U thr r\. ital.ilil y ..f thr nerve CM 1
•11; the s. i.iti, ; . \c preparation in a small gas chain)'
let, whiih arr applird t«> thr proximal and distal extremities of the
'• MindiiitiriK rthrr v.ipor- into thr < hamltcr : ility disappear-
end of the nerve \vhi< h i- larthr-t irmovrd from the muscle.
148 A LABORATORY GUIDE IN PHARMACOLOGY
Experiment 9. (Demonstration) Anesthesia of Nerve by Freezing. —
Decapitate a pithed frog and trim away the viscera so as to expose the
sciatic plexuses. Expose the sciatic nerve of one thigh, without cutting or
injuring it, and support it on a match-stick. Lay the frog with the ventral
surface upward, arrange electrodes on the plexuses, and see that a weak
stimulation is effective (flexing the knee before stimulating). Freeze the
exposed sciatic by a spray of ethyl chlorid. The leg will make some spon-
taneous contraction during the freezing, but in a short time it will cease to
respond to the electric stimulation of the plexus, the conductivity of the
nerve being paralyzed. Remove the spray and melt the nerve by the heat
of the finger: the stimulation again becomes effective after a time.
Experiment 10. (Optional) Anesthesia of Skin by Freezing. — Spray some ethyl
chlorid on the back of the hand: this produces pain and then anesthesia.
Experiment u. (Optional) Intravenous Cocain Anesthesia. — Into an ear vein of a
rabbit inject cocain, 10 mg. per kg. (i c.c. per kg. of i per cent.), noting the time of injec-
tion.
Observe the motor symptoms: how soon the animal becomes quite paralyzed. Ob-
serve also the anesthesia toward pinching or pin pricks. Note the respiration. Is con-
sciousness lost?
Record the time of onset and the duration of the anesthesia.
Question. — What would be the objections to using cocain intravenously on patients?
Experiment 12. (Optional) Infiltration Method of Anesthesia. — ("Quaddel" method
of Braun.) Wash the skin of the flexor surface of the forearm with alcohol. With a
sharp and strictly sterile hypodermic needle introduced into (not under) the skin, parallel
to the surface and just far enough so that cannula-opening is well covered, inject slowly
a drop of the sterile solution, so that a small wheal (split-pea size) is formed. Test the
sensibility to needle-pricks immediately after injection and in five-minute intervals.
The solutions should be made with normal saline and warmed. They may be started
with concentration of 0.025 Per cent. A number of tests can be made in close succession.
CHAPTER XXXIH
MUSCULAR CONTRACTION: SKELETAL MUSCLE, CILIA
Explanatory. — The actions of drugs on striped muscle are scarcely util-
ized in therapeutics, but they help to explain the effects on the cardiac
muscle, which are very important. They are also of considerable scientific
interest. The effects may involve the form of the contraction curve, its
height, the rapidity of contraction or of relaxation, the load which the
muscle can lift, the total work which it can perform, the promptness of
fatigue, the minimal effective stimulus, the latent period, the rate of stimu-
lation required for fatigue, etc. As a general rule, these functions are all
affected in the same sense.
The majority of muscle poisons may be arranged in three groups, which
are illustrated typically by caffein, quinin, and veratrin.
Ca/ein increases the activity of the muscle in small doses; larger doses
produce phenomena analogous to fatigue. Very large doses throw the
muscle into rigor. The methyl-xanthins (caffein, theobromin, etc.) are the
only typical representatives of this group.
Quinin depresses the muscle, and finally paralyzes it, without producing
rigor. Only the smallest doses are somewhat stimulant. All protoplasmic
poisons and apomorphin and potassium, calcium, and metallic salts produce
these effects.
CHAP. XXXIII MUSCULAR CONTRACTION! SKELETAL MUSCLE, CILIA
I49
I 'cratrin causes the muscle to remain contracted for a considerable time,
the curve resembling somewhat that of tetanus. It can be distinguished
from this by the secondary contraction (see Exercise II, Experiment 2, V);
it is, however, an active contraction, for the muscle can sustain a weight.
The effect is lessened by all agents which depress the muscle.
In studying the effects of drugs on skeletal muscle, they may either be
injected into the lymph-sac or into the aorta; or the muscle may be laid
in a solution of the drug in normal saline. Special conditions determine
which of these methods is to be preferred. When the muscle is laid in the
solution the drug is not always rapidly absorbed. It may therefore happen
that one muscle will be scarcely affected by a strong solution, while a weak
solution may produce severe effects in another preparation. All the
muscular poisons act equally well after curare, showing that their action is
indeed exerted directly on the muscle cells.
(Copies of the tracings should be inserted in the note-books.)
Technical References.— Experiments on Muscle and Nerve.— Stewart, 780; Tiger-
stedt, 2.3, 187; Robert, Intox., i, 168.
Principles of Registration. — Tigerstedt, 1.4, 51; Photographic Registration, ibid., i.i,
65; 1.4, 25.
FJcctrophysidogy. — Ibid., 2.3, 317; Stewart, 814; Electrometers and Galvanometers,
Hgentedt, 2.3, 419; string, ibid., 428; Current of Rest, Abderhalden, 3, 551; Action Cur-
rent as Index of Glandular Activity, Cannon and Cattell, 1916, Amer. Jour. Physi»l.,
4i, 39-
Fig. n. — Arran^rmcnt for muscle tracing.
Technical Notes.— Tracings from Kxdsc,/ (nistn><n<-niins Muscle.— The
muscle is attached by hooks or >trini:>, as shown in ML:, i i. The attach
mem to the lexer i- be>t made with a bent pin. ><> that the point of attaeli
ment, and thereby the excursion, can be altered as needed. A weight of
about logm. should be suspended on the other limb, about an e<|iial distance
the fulerum. The nerve may be laid on the electrode-. 1 1 the muscle
be -timulated direetly. tine wires, connected with the secondary eoil.
iru>t direetly through the mu-c le.
ed unle-> the nniM le i- to be tetani/ed. The
lever i> adjusted at a tangent to the drum until it traces easily when the
lever Is moved . The writing-point ihoukl be bent d the drum. The
faste Q| the drum i-> needed to .\///>7r the form <>t < otilr.i, t:
a -oluti. ted by plaeim: it in a b. test-tube,
and rai>inur this so as to immerse the muscle (Fig. n).
150 A LABORATORY GUIDE IN PHARMACOLOGY
A time record may be placed on the tracing by a writing-point attached
to a vibrating tuning-fork.
Similar arrangements are described in Heinz, i, 434.
Musck Tracings from Intact Frog. — Fuehner, 81.
Muscle Levers. — The substantial pattern shown in Fig. 12 has proved
very satisfactory. The muscle is stretched between the arm of the lever
and the rod (b) which is set into (a) by a thumb-screw. The levers are
prolonged by a narrow strip of aluminum.
L-shaped Levers are required for a horizontal pull.
Other types of levers are described, for instance, in Tigerstedt, 1.4, 17.
For writing points one may use tapering bits of parchment paper 5 cm. long and i cm.
wide at the base. These are attached to the end of the straw-levers, etc., by sealing wax
or colophonium cement. Points of celluloid, aluminum or steel, or the blunt end of a
needle, can be similarly used. The end of the writing-point should be bent slightly toward
the drum. It should be placed at a tangent, pointing in the direction toward which the
drum is moving.
Stands for Supporting Levers, etc. — A rather short stand with heavy semicircular
base (Harvard) is best. It is furnished with double clamps ("mouffen").
An adjustable stand is very convenient if great accuracy of adjustment is needed.
This is secured by a micrometer screw.
Kymographs (Drums). — Movements are registered as "tracings" on cylinders moved
by clockwork or motors. The ordinary Harvard kymograph answers for pharmacologic
work.
By using two drums (or the device described by McPeek, Jour. Amer. Med. Assoc.,
61, 2065, 1913) a longer record can be secured; but this is not necessary if some extra
cylinders are smoked in reserve. D. E. Jackson (Jour. Amer. Med. Assoc., 56, 1705, 1911)
describes a spinning device for faster speeds. Other types are described in Tigerstedt,
1.4, i; Pittenger, 1913, Jour. Amer. Pharm. Assoc., 1498, etc.
Speed of Kymographs. — Three speeds are needed, approximately: 5 to 10 cm. per
second for muscle tracings; 10 cm. per minute for details of blood-pressure, etc.; and
2 cm. per minute for prolonged blood-pressure respiration, etc.
A very rapid speed may be secured by raising the drum from the clockwork and
spinning it like a top by a weight attached to a cord which is wound about the drum.
The tracing should be taken immediately after the weight has fallen.
Tracing Paper. — The cylinder of the drum is covered with paper on which the record-
ing instrument writes. The paper is drawn snugly around the drum, the free edge of the
paper being pasted with mucilage on to the first layer. Superfluous paper is trimmed
off. The writing may be done with ink from a small glass feeding tube attached to the
writing instrument. A more generally useful method, however, is to use a paper with
glazed surface and covered with a thin layer of soot, on which the levers, etc., trace.
Dextrin Mucilage (Sykes). — Mix 180 gm. of dextrin with 180 c.c. cold water; add 240
c.c. boiling water and boil five minutes, stirring constantly. Add hot water q. s. 400 c.c.
When cold, add 30 c.c. dilute acetic acid, 10 drops phenol, and 30 c.c. of glycerin, pre-
viously mixed.
Smoking the Drum. — A uniform layer of soot is deposited on the paper by revolving
the drum rapidly in the flame of a fish-tail burner. A stand for supporting the drum
while it is being revolved and smoked can easily be constructed from a small box.
A blacker soot may be obtained by passing the gas through a wash-bottle containing
a mixture of equal parts of benzin and benzol.
Starting the Tracing. — The tracing is always started where the paper
joins; and in detaching it from the drum it is cut along this line.
Abscissa. — It is generally advisable to trace an abscissa on the drum
by revolving it against the writing-point before the actual tracing is started.
With muscle tracings the abscissa should be at the point of rest; with
blood-pressure tracings it should be at the zero level. This abscissa may
be used for marking signals and time.
CHAP. XXXIII MUSCULAR CONTRACTION: SKELETAL MUSCLE, CILIA
.'. — Muscle lever, actual sixe
Signal MaRnfl. — Thr Dimple II.it\.inl t\|.r suffices.
••l-prrxxurr • itty-second
period with llu- -icn.il after the drum is running smoothly . and
especially with fast speeds, a cl<>< k. MI. h u tlu li.ir\.ini. may be used. -. also
be used as signal by I..IHU-. ting an extra key which short « it, UIN tin
152
A LABORATORY GUIDE IN PHARMACOLOGY
The additional keys may be disposed at various points convenient to the operator. Locke,
1908, Quart. Jour Exp. Physiol., i, 359, describes a system of multiple signals with a
single lever. Haie, 1916, Jour. Pharmacol., 8, 445, describes a modification of the
Harvard time-recording apparatus.
Notes on Tracing. — It is distinctly advantageous to transfer to the
tracing all the notes which have been taken during the experiment. This
may be done without confusion by numbering the signals to correspond with
the notes. The writing on the smoked surface is done with a blunt needle
or dry pin, after taking the paper from the drum and before varnishing.
A marking board is very helpful to avoid
disfiguring the tracing. The tracing is laid
on a board over which another board slides
on runners.
Varnishing. — The marked tracing is
passed through shellac varnish and hung to
dry.
Varnish Trough. — Waste of varnish can be pre-
vented by the device shown in Fig. 14. By stepping
on the treadle the reservoir is raised so that the var-
nish flows into the trough. When the treadle is re-
leased the reservoir descends and the varnish flows
back.
A portable varnish fixture is described by Hos-
kins, 1916, Jour. Amer. Med. Assoc., 67, 874.
Varnish. — This is made by dissolving orange
shellac in 15 parts of alcohol and decanting.
Blue prints of tracings may be made by laying the
tracing on a sheet of sensitive blue-print paper, cover-
ing with a plate of glass, exposing to sunlight for a
day, and washing.
Clock
Fig. 13. — Diagram of time and simple signal.
Fig. 14. — Varnish trough,
Lantern-slides of Curves. — Straub, 1913, Zs. biol. Tech., 3, 267.
Demonstration of Tracings. — Tracings may be demonstrated by placing them before
a light. An efficient lantern for this is made by a box the size of the tracings, open in
front, lined with asbestos, and containing three incandescent lamps. The front of the
lantern is closed by two plates of glass, one in front of the other, between which the trac-
ing is slipped. With long paper kymographs the lamps may be hung between the cylinders.
CHAP. XXXIII MUSCULAR CONTRACTION: SKELETAL MUSCLE, CILIA 153
EXERCISE I.— (ALL GROUPS) FORM OF CONTRACTION CURVE
(REPORTER I, B)
Arrange apparatus for muscle-tracing (Tech. Notes) with fastest speed
of drum. Arrange induction coil for single break shocks (Tech. Notes,
page 136).
Make a muscle preparation of the gastrocnemius, with a bit of femur
attached (Tech. Notes, page 137). Tie it on the muscle-lever arranged
for immersion in beaker (see Fig. n). Pass fine wire electrodes from sec-
ondary coil into muscle. Immerse muscle in normal saline for five minutes,
and make two or three fast tracings of single muscular twitch, single-break
shock.
Remove the beaker and replace the saline solution by the drugs named
below. Take a very slow tracing, without stimulation, during the course
of the immersion, and a fast tracing, with single-shock stimulation, at inter-
vals of five minutes. Where different strengths of solution are to be used
they may be changed every five minutes or so. The different groups may do
tin- following experiments:
(Group I) Caffein. — Solution in N. S.1 of i : 10,000; then i : 1000; then
i : 100. The more dilute solutions cause a higher contraction, with little
change in the form of the curve. Stronger solutions produce a lengthening
of the relaxations. The curve then becomes lower, the contraction is slower,
and with the strongest solution the muscle does not contract at all. With
fairly strong solutions the relaxations may show a series of waxes, which
are not yet satisfactorily explained.
(Group II) Theobromin-sodium Salicylate. — Solutions in N. S. of i :
10,000; i : 1000; i : 100.
(Group III) Quinin-hydrochlorid. — Solutions in N. S. of i : 10,000;
i : 1000; i : 100. The weakest solutions may increase the height of con-
traction somewhat; but even fairly weak solutions lower the contraction, and
finally paraly/.e the muscle completely.
(Group IV) Potassium Chlorid. — Solutions in N. S. of i : 10,000 and
i : 1000: depression.
(Group V) Alcohol. — Solutions in N. S. of i : 1000; i : 100; i : 10:
depression. The weakest solution may stimulate somewhat.
(Optional) Chloroform or Ether may be applied as vapor in a gas-chamber.
QUESTIONS
Enumerate the dru^s which increase the height of contraction; thoe
which lower it; and th<»c which have both effects, according to concentra-
tion.
EXERCISE H.— (ALL GROUPS) VERATRIN EFFECT
«TB II. B)
Arrange apparatus as I'm- I • el, but with fairly -low drum (about I
iiH h per minute). CrOUDS I, HE, and V : Inject 0 i : 10,000 veratrin
into the lymp I fn>u'. When it -how- the typical
QWOn <»f l'M> "H jumpii. mu-< le |>: D and take a -low i
1 1 the typical action ha- hern reached, the height and rapidity of the
> N. S. stands for normal saline *»l
154 A LABORATORY GUIDE IN PHARMACOLOGY
contraction is normal, but the relaxation is greatly prolonged. Give the
second gastrocnemius to Groups II or IV.
(Optional) The veratrin effect may also be obtained by immersing a thin muscle (the
sartorius) in veratrin solution, i : 1,000,000 to i : 100,000 in X. S.; repeating the stimula-
tion every five minutes until a typical t racing is obtained.
The different groups use the veratrinized muscle for the following experi-
ments:
(Group I) Incipient Fatigue. — Stimulate the muscle every five seconds,
taking a slow continuous tracing: the relaxation shortens to normal, even
before the height of the contraction is lowered.
(Group II) Temperature. — Place the muscle in N. S. solution which has
been kept on ice. Note the temperature and obtain a tracing. Raise the
temperature, immersing the beaker in hot water, so that it takes about five
minutes to rise to 10° C- Take another tracing. Continue to raise the
temperature, 5 degrees per minute, taking tracings at 15, 20, 25, 30, and 35
degrees. The lower temperatures lessen the contracture; 20 and 30 degrees
prolong it; 35 degrees lessen it. (If the veratrin action is only slight, the
contracture may appear increased by cold, for this prolongs the relaxation
in unpoisoned muscle.)
Heating and Cooling the Muscle. — The muscle may be heated or cooled by laying
it in normal saline solution of the required temperature. Better results can be ob-
tained by surrounding the muscle with a box containing water at the proper temperature
(Harvard muscle warmer).
(Group III) Potassium.— Add KC1 i c.c. of i per cent, per 10 c.c. of vera-
trin solution, and stimulate at intervals: the relaxation is shortened.
(Group IV) Ether. — Add a few drops of ether to the veratrin solution, and
stimulate from time to time: the relaxation is shortened.
(Group V) Secondary Contraction. — Make a muscle-nerve preparation
from a normal frog. Lay the nerve of this on a good veratrin preparation,
so that the cut surface lies on the tendon, and the long surface of the belly
of the veratrin muscle. The nerve should be raised between the two points
of contact by a match-stick. Stimulate the nerve of the veratrin muscle
with a single break shock: the current of action will stimulate the normal
muscle, so that it will also contract; but the contraction will be short, whereas
the contraction of the veratrin muscle is prolonged. This shows that the
veratrin contraction is not a tetanus; for if it were, the normal muscle would
also remain contracted. Convince yourself of this by stimulating the nerve
of the veratrin muscle with the tetanizing current: the normal muscle now
remains contracted.
QUESTIONS
(a) Describe the veratrin effect.
(b) How may this be antagonized?
(c) What do these measures have in common?
(d) How is it proved that the veratrin curve is not a tetanus?
EXERCISE HI.— (DEMONSTRATION OR GROUPS I AND II) MAXIMAL
LOAD (ISOMETRIC CONTRACTION)
(REPORTER I, A)
Make two muscle preparations. Determine the lifting power as de-
scribed below. Lay one muscle in N. S., the other in the solutions. De-
CHAP. XXXIII MUSCULAR CONTRACTION : SKELETAL MUSCLE, CILIA 155
termine the maximal load every five minutes, transferring the poisoned
muscle to solutions of increasing concentration.
Experiment i. (Group I) : Use Caffein, i : 10,000; then i : 1000 (in N. S.).
Experiment 2. (Group II): Use Quinin, i : 10,000; then i : 1000 (in N.
s.).
The Lifting Power of a Muscle. — A convenient apparatus for studying
this consists in a stiff straight brass wire (4 mm. diameter), about 6 inches
long. One end of the wire is securely clamped to a stand; the other is
prolonged by a straw, to exaggerate the movement. A stiff iron rod (J inch
diameter, 6 inches long) is clamped on the same stand, 3 inches above and
parallel to the brass wire. The muscle is tied to the two rods so that it may
be moved toward or away from the stand. The nearest point to the stand
i> noted at which stimulation of the muscle causes a perceptible movement
of the lever. This will be the nearer, the greater the lifting power of the
muscle.
Another method is as follows: The muscle is connected with a Harvard muscle lever,
\\hirh is supported by the after-load screw. A weight-pan is suspended from the lever
at the point where the muscle is attached, and weights are added until the muscle is just
unable to move the lever when stimulated.
QUESTION
What are the effects of quinin and of caffein on the lifting power of
muscle?
EXERCISE IV.— (DEMONSTRATION) FATIGUE
(REPORTER I, A)
Make two muscle preparations. Immerse one in the poison solution,
the other in normal saline for five minutes. Obtain a tetanus tracing (Tech.
NOtes, Chap. XXXII) first from the poisoned muscle; then just under this
< .11 the drum, from the saline muscle. Use the same slow speed of drum, and
the same strength of stimulation, for both tracings. Note which fatigues
the more quickly.
Experiment i. (Group III): Use Caffein, i : 10,000 in N. S.
Experiment 2. (Group IV): Use Quinin, i : T 0,000 in N. S.
Experiment 3. (Group V): Use Alcohol, i : 100 in N. S.
tion and amputation of one leg increases the resistance of the other leg to
fatigue (Criderand Robinson, 1916, Amer. Jour. Physiol., 41, 376).
QUESTION
Describe the effects of these drugs on the hit inability of the muscle.
EXERCISE V.— (OPTIONAL)
Action of Drugs on Fatigue in Man (Optional). Thi> may be studied \>\ the -prin^
ergograph. A normal tracing is taken and this is repeated at half-hour intervals
-0.3 Km. «>f ..tlirin or ;o to 40 < .c . of ^oper.eni. al.ohol. Some pnulin- is re<|uired
•<• reliable results can be obtained.
EXERCISE VI.— (DEMONSTRATION OR ALL GROUPS; OSMOTIC EFFECTS
ON MUSCLE AND NERVE
III. Hi
I)irn! OOntad uith \\ati-r poisOOl mu-< lr. partly \>> < absorp-
tion <»f wain. partly l»y tin- \\ithdra\val uf >alt>. These act it m> a if l;r
156 A LABORATORY GUIDE IN PHARMACOLOGY
due to osmosis. Strong salt solutions cause irritation and eventually paral-
ysis by withdrawal of water.
Experiment i. (Group I) Excitability. — Make two muscle-nerve prepara-
tions. Use the arrangement described in Chapter XXXII, Exercise IV,
Experiment 4. Immerse the nerve of one and the muscle of the other in
tap-water, and observe the loss of excitability from time to time.
Note whether there are any muscular twitchings.
The excitability of a muscle or nerve is observed very simply by noticing the greatest
distance or angle of the secondary coil which will just give a contraction (single break
shocks). Care must be used that the electrodes make good and equal contact.
To compare the effect of a drug on muscle and nerve, two muscle-nerve preparations
are made from the same animal. A microscopic slide is placed in an evaporating dish so
as to form a bench, and the bottom of the dish is filled with the solution (which should not
touch the slide). The two preparations are now arranged so that the nerve of one and the
muscle of the other are in the solution, while the muscle of the first and the nerve of the
second lie on the bench, /. e., outside of the solution.
Questions. — (a) What are the effects of water on the excitability of muscle
and nerve?
(b) Which is more susceptible?
Experiment 2. (Group II) Water Rigor. — Suspend a thin strip of muscle
(the sartorius) of frog so that half of it dips into water: this will be seen to
become thicker and shorter.
Questions. — (a) Why does the muscle swell in water?
(b) In what way does this affect its functions?
Experiment 3. (Group III) Water Rigor Contracture. — Take slowest
speed tracing of gastrocnemius immersed in water (without stimulation).
This shows shortening. Determine the weight required to stretch the
muscle to its original size.
Questions. — (a) Is the muscle in water-rigor able to sustain a weight?
(b) How does this compare with rigor?
(c) How is the difference explained?
(d) Why does the muscle shorten in swelling?
Experiment 4. (Demonstration) Perfusion with Water. — Decapitate a
frog, leaving lower jaw. Divide one sciatic plexus. Insert cannula into
descending aorta and wash out the blood with saline. Suspend frog by jaw
and attach one foot to light lever.
Perfuse vessels with water: in a short time the muscles will show fibrillary
twitchings and these will be succeeded by general convulsions. Eventually
there is paralysis.
Questions. — (a) Are the twitchings of central or peripheral origin? Why?
(b) Is the action on the nerve-trunk? (compare Experiment i).
(c) Is the action probably on the nerve-endings or on the muscle? Why?
(d) How could this be definitely decided?
Experiment 5. (Groups IV and V) Hypertonic Solution on Nerve. —
Arrange a muscle-nerve preparation on a lever, writing on a slow drum.
Let the nerve dip into 10 per cent. NaCl solution. The muscle will
execute a series of contractions, then remain in tetanus, and finally go into
paralysis.
Questions. — (a) How does the salt solution act on the nerve?
(b) How could you show that the effect is not due to the NaCl as such,
but to the withdrawal of water?
CHAP. XXXIII MUSCULAR CONTRACTION I SKELETAL MUSCLE, CILIA 157
EXERCISE VII.— (ALL GROUPS) RHYTHMIC CONTRACTIONS OF SKELE-
TAL MUSCLE iBARIUM, CALCIUM, DECALCLFICATION)
(REPORTER IV, B)
Disturbance of the ratio of ions about a muscle, as by administration of
Barium, by the abstraction of Calcium with citrate or fluorid, etc., brings
out the rhythmic functions which are inherent, though latent, even in skele-
tal muscle. Restoration of the ion's again allays these contractions.
Musck-s vary greatly in the facility with which rhythmic contractions
are induced.
Experiment i. (Group I) Citrate and Calcium. — (a) Arrange a frog's
muscle on a lever, writing on a slow drum. Immerse the muscle in a beaker
of 5 per cent. Sodium Citrate. Rhythmic contractions will appear within a
few minutes.
(b) Transfer to Calcium Chlorid i per cent, in normal saline. When the
contractions have ceased, again place in the Citrate and see whether they
reappear.
Experiment 2. (Group II) Citrate and Barium. — (a) Same as Experi-
ment i (a).
(b) Transfer to i per cent. Barium Chlorid in N. S.: the contractions
become stronger.
Experiment 3. (Group III) Barium and Calcium. — (a) Take tracing
from muscle immersed in i per cent. Barium Chlorid in N. S.: rhythmic
contractions.
(b) Add an equal volume of i per cent. Calcium Chlorid solution: the
contractions are not allayed.
Experiment 4. (Group IV) Citrate and Potassium. — (a) Same as
Experiment i (a).
(b) Transfer to o.i per cent. Potassium Chlorid in N. S.: the contrac-
tions are allayed.
Experiment 5. (Group V) Fluorid and Calcium. — (a) Same as Experi-
ment i (a), but using 0.5 per cent. Sodium Fluorid in place of the Citrate.
(b) Same as in Experiment i (b).
QUESTIONS
(a) Why does the cardiac muscle normally contract automatically and
rhythmically, and the skeletal muscle only on stimulation and then by a
single twitch?
Why are the contractions in citrate and fluorid attributed to tin-
withdrawal of calcium rather than to a direct action of the citrate or tluorid?
(c) How U it >ho\vn that the presence of calcium is not sulVu icnt to pro-
duce the calcium effects, but that it must be ionized? (Consider Experi-
ment
(d) Has the production orallayanee of rhythmicity any definite relation
to the valence of tin i
(e) Has the act ion «.f K in I.xperimcnt 4 any relation to the Calcium ions?
How could this be >hown?
(/) Could the action of calcium be simply that of a depressant? (Con-
sider Kxpcriment 3.)
158 A LABORATORY GUIDE IN PHARMACOLOGY
EXERCISE VHI.— (DEMONSTRATION OR ALL GROUPS) VITALITY OF
TISSUES INFLUENCED BY SALTS
(REPORTER V, B)
Excise the hearts of the frogs used in other experiments, and place
in watch-glasses with the solutions named below. Note how long they con-
tinue to beat:
(Group I) : Ringer's Solution.
(Group II) : Ringer's Solution without Ca.
(Group III) : Ringer's Solution without K.
(Group IV): Ringer's Solution, triple strength.
(Group V) : Distilled water.
QUESTIONS
(a) Tabulate the solutions in the order in which the hearts stop.
(b) Why does the withdrawal of Ca or K injure the heart?
(c) Why is the heart injured by triple strength Ringer's solution?
(d) Why is it injured by water?
EXERCISE IX.— (DEMONSTRATION) PROTOPLASMIC DEPRESSANTS
(REPORTER V, B)
Explanatory. — These paralyze nervous and muscular structures, but
differ from the muscle-nerve poisons by acting also on monocellular organ-
isms, and often even on ferments. They can be observed conveniently on
ciliated cells and on vegetable seeds.
Experiment i. Paralysis of Cilia. — (a) Cut off the lower jaw of one of
the frogs used in a former experiment so as to expose the ciliated mucosa of
the pharynx and esophagus. Irrigate with normal saline solution. De-
termine the time which a small bit of cork requires to travel a certain dis-
tance (which may be marked off by pin-pricks). Take a number of ob-
servations, keeping the mucous membrane moist. Irrigate with the ether
solution, and after a few minutes repeat the observations. It will be found
that the ciliary movement is greatly slowed or arrested. If the cilia have
not been too profoundly injured they may recover if they are thoroughly
washed with normal saline solution.
(b) (Optional) The ether may also be administered in vapor form by
supporting the esophagus on a small stand in a tumbler, which contains a
little cotton saturated with ether, arid which is covered by a glass plate.
A recording arrangement (cilio^cribe) is described by Dixon and Inchley,
1905, Jour. Physiol., 32, 395.
Experiment 2. (Optional) Germination of Seeds. — "Arrange two 8-ounce wide-
mouth bottles with stoppers fitted with glass tubes, letting one tube extend to near the
bottom of the bottle. Suspend in each, by means of cotton, a dozen seeds — corn, wheat,
clover, beans, etc. — and introduce just enough water to maintain a saturated vapor.
Set both bottles in a window. Through one pass ether vapor, through the other air, twice
a day for a week. The seeds in both will swell from the absorption of water, but only
the bottle with pure air introduced will grow. Reverse the two. The sprouting grain
will have its growth checked and the etherized seeds will begin to grow" (C. W. Greene).
QUESTIONS
(a) What are the effects of ether on cilia?
(b) On germination?
(c) Is the "narcotic" action of ether confined to the nervous system?
CHAP. XXXIV SMOOTH MUSCLE: INTESTINE, UTERUS, AND ARTERIES 159
EXERCISE X.— (DEMONSTRATION OR ALL GROUPS) ASTRINGENTS
(REPORTER I, F)
Astringents precipitate proteins, thereby diminishing their affinity for
water. The tissues, therefore, shrink or contract when exposed to astrin-
gents. The astringent power can be demonstrated and compared as in the
following experiments.
Technical References. — Dreser, 1908, Arch, internal. Pharmacod., 18, 114; Fuehner,
138; Heinz, i, 126.
Experiment i. (Group I) Astringent Action on Lung (Dreser's Method).
— (a) Carefully dissect out a fro«j lung with its bronchus and insert a small
cannula into bronchus (keep lung moist with water). To free end of
cannula attach a 2-c.c. pipet divided into -fa c.c.
Insert free end of pipet into a 2oo-c.c. graduate to a depth corresponding
to 50 c.c. and read height (from the cross lines on graduate) that water
ascends into the pipet.
Repeat observations at different levels — 100, 150, and 200.
Now place lung in a i per cent. Tannin solution for two minutes and
repeat above observations.
(b) With the other lung make similar observations with wrater and i
per cent. Silver Nitrate.
(Group II) : Do (a) as above; in (b) use i per cent. Zinc Sulphate
(Group III): Do (a) as above; in (b) use Copper Sulphate.
Experiment 2. (Groups IV and V) Astringent Action on Mucous Mem-
brane.— Cut a strip of mucous membrane as long as possible from the
mouth of a frog. Attach to a lever and immerse in N. S. ; let it trace on a
slow drum, first marking a base line. Add to the N. S. the following drugs,
and note whether the tracing shows a contraction:
(Group IV) : i c.c. of 10 per cent. Tannin per 10 c.c. of N. S.
(Group V): i drop of Epinephrin, i : 1000, per 10 c.c. of N. S.
QUESTIONS
(a) Arrange the astringents in the order of their efficient
(b) In what conditions would this action be useful?
(c) Is epinephrin a true astringent?
CIIAI'I I U \X\IV
SMOOTH MUSCLE: INTESTINE, UTERUS, AND ARTERIES
Explanatory. Tin- properties «>i" -mouth IIHIM le differ in essential re
u fnmi thos(. «.i" striped IIUIM le. They an- affected in a rather Ip
manner by the autonomic poisons acting on their muscle substance, on
the myMiieiiral jiuietinn. <»r MII the gangUOD iell>. The analysis of these
phen< red later.
The niMst important smooth mu-rle System are tlioM- of the gastro-
t the uterus, the l>rom hioles, and the arteries.
The phenomena can be most conveniently studied and analyzed on
excised mammalian tissues, bathed in warm Locke's fluid, through which a
l6o A LABORATORY GUIDE IN PHARMACOLOGY
constant stream of oxygen or air is passed. The muscles may be attached
to levers and tracings obtained, just as with skeletal muscle.
Effects of Drugs on Peristalsis. — Drugs which increase peristalsis may
be grouped as cathartics; those which diminish peristalsis as antidiarrhcvica.
While peristalsis and especially defecation are to some degree controlled
by the central nervous system, almost all the drugs which influence them
act peripherally. The remedies which are utilized therapeutically to in-
fluence peristalsis are mainly direct irritants, chemic or mechanic, or as-
tringents. These act only when they are introduced into the alimentary
canal.
Peristalsis may also be influenced by peripherally acting muscle-nerve
poisons. They are rarely used in therapeutics (except in intestinal paresis),
because their effects are not confined to the intestinal tract; but they are of
considerable importance in toxicology and pharmacology.
The peristaltic movements are arrested by atropin or epinephrin, stimu-
lated by muscarin, physostigmin, pilocarpin, and nicotin. There is a
mutual antagonism between atropin on the one hand and muscarin, pilo-
carpin, and physostigmin on the other, the effect corresponding to which-
ever drug is present in excess. Atropin prevents the effects of nicotin, but
not vice versa. Barium is active after atropin, but not the reverse.
TECHNICAL NOTES
Decerebation of Mammals. — This is employed when it is desired to exclude disturbing
cerebral effects or anesthesia.
Sherrington's operation for cats is as follows (Jour. Physiol., 1909, 38, 375; van
Leeuwen, 1913, Arch. ges. Physiol., 154, 306; Forbes and Sherrington, Amer. Jour.
Physiol., 35, 367) :
"The animal (cat) being deeply anesthetized with chloroform, a cannula is inserted
into the trachea. Both common carotids are ligated. A transverse incision through the
skin is made over the occiput and extended laterally close behind the pinnae. The skin
is retracted backward so as to expose the neck muscles at the level of the axis vertebra.
The ends of the transverse processes of the atlas are then felt for and a deep incision made
through the musculature just behind these processes. The large spinous process of the
axis is notched with the bone forceps. A strong thick ligature is passed by a sharp-ended
aneurysm needle close under the body of the axis, and is tied tightly in the groove left by
the incision behind the transverse processes of the atlas and the notch made in the spinous
process of the axis. This compresses the vertebral arteries where they pass from trans-
verse process of axis to transverse process of atlas. A second strong ligature is then
looped round the neck at the level of the cricoid, and is so passed as to include the whole
neck except the trachea. Decapitation is then performed with an amputating knife
passed from the ventral aspect of the neck through the occipito-atlantal space, severing
the cord just behind its junction with the bulb. The ligature round the neck is drawn
tight at the moment of decapitation. The severed head of the deeply narcotized animal
is then destroyed. Hemorrhage is extremely slight. If there is oozing from the verte-
bral canal it is arrested by raising the neck somewhat above the rest of the carcase. The
carcase is placed on a small metal-topped table warmed by an electric lamp below. Arti-
ficial respiration is employed to ventilate the lungs, the fresh air supplied from the bellows
being warmed by passing through a chamber containing a small electric lamp. The
skin-flaps are stitched together, covering the exposed end of the spinal cord and other
structures bared by the amputation wound. The carcase will continue for several hours
to exhibit good reflexes employing the skeletal muscles, although the arterial blood-
pressure is low, often not more than 80 mm. Hg. Reflexes on the arterial blood-pressure
are usually obtainable, but are poor. The rectal temperature is fairly well maintained
if the table and air from the bellows be suitably warmed; it can easily become too high
if the table be overwarmed.
"The execution of the whole procedure occupies about six minutes."
The operation transects the cord about 4 mm. behind the calamus.
It is well to wait for one-half hour to allow the anesthetic to disappear.
The brain may also be cut with a spatula through a trephine opening (Magnus, Arch,
ges. Physiol., 130, 254, 1909); spinal animals are not subject to shock by subsequent divi-
CHAP. XXXIV SMOOTH MUSCLE: INTESTINE, UTERUS, AND ARTERIES l6l
sion of the cord at lower levels. The dcccrebration of dogs is described by Sherrington,
1909, Quart. Jour. I': 115.
The central nervous system may also be excluded by the injection of o//, etc., into its
circulation (Tigerstedt, 3.4, 55); and by the closure of the arteries supplying the brain
(Stewart, Guthrie, and Pike, 1906, Jour. Exp. Med., 8, 289); Guthrie, 1911, Zs. biol.
•!., :>. 138); Langley, 1912, Jour. Physiol.. 45, 239, secures partial blockage of the fore-
brain by the injection of starch suspension into the peripheral end of the right carotid
artery.
Decerebration of Rabbits for Survival Experiments. — Morita, 1915, Arch. exp. Path.
Pharm., 78, 188.
EXPERIMENTS ON PERISTALSIS IN INTACT ANIMALS
A discussion of the technic is given by R. Magnus, Ti<:erstedt's Handbuch, 2.2, 115,
1911; also Abderhalden, 6, 604; Robert, Int<>\.. i. 250. Additional methods by Hallion
and Nc-ttiT ((.'. R. Biol., 182 and 254, 1907 — balloon method); Alvarez, 1915, myograph,
Anier. Jour. Physiol., 37, 267; Joseph and Mcltzer, non-anesthetized animals (Soc. Exp.
Biol. Med.. ;. 95, 1910); Trendelenburg (Zs. Biol., 61, 67, 1913).
Gastric Movements. — Tigerstedt, 2.2, 99.
Hunger Contractions. — Carlson, Amer. Jour. Physiol., 33, 95.
Operations on Digestive Tract. — London in Abderhalden, 3, 76.
Digestive Fistula.— AhderhaMen, 6, 564; Thiry-Vella, 6, 466.
Digestive Experiments on Animals. — Zunz in Abderhalden, 3, 122.
Collection of Digestive Secretions. — Ibid., 189.
Digestive Tract of Frog. — Robert, Intox., i, 187.
Examination of Stomach Contents.— Abderhalden, 8, 44.
Relative Weight of Gastro-intestinal Tract of Rabbits. — Livingston, 1914, Jour.
Exp. Med., 19, 339.
Blood-supply of Stomach.— Burton-Opitz, 1910, Arch. ges. Physiol., 135, 205.
EXERCISE I.— (DEMONSTRATION) PERISTALSIS OF EXPOSED INTES-
TINES; NICOTIN ON GANGLIA
(REPORTER II, F)
Use a decerebrated rabbit (Tech. Notes). Stretch on board; make small
incision in linea alba, and draw forth a loop of small intestine.1
Experiment i. Bayliss-Starling Reflex. — Observe that pinching with
forceps causes a spreading peristalsis (mechanical stimulation)} the intestine
contracting above the stimulus and relaxing below.
Experiment 2. Local Irritation. — Apply a crystal of salt: spreading
stimulation.
Experiment 3.— Apply at another place a few drops of -^ per cent.
pkysostigmin: local constriction (stimulation of muscles and endings).
Experiment 4.— Apply at another place a drop of i per cent. BaCl«:
strong conMric tion f simulation of muscle).
Experiment 5. Apply at another place a drop of -jV per cent, atropin:
pcri-taUi- < 68
Experiment 6. Colon Peristalsis. — Pinch the ascending colon, or apply
-olution of barium chlorid: a tonic contraction ring occurs, and
from tlii- Mart- an amending peri-taUi-.
Experiment 7. Nicotin on Ganglia and Nerve-fibers. — Expose the supe-
rior cervical ganglion of rabbit. Stimulation causes constriction of tin-
ear ve— els and dilatation of the pupil. Paint i per cent, nicotin on tin-
nerve below the ganglion. A >timuhi- applied central to thi- point is -till
:i\e. showing that tin- nerve fibers are not paralyzed by the poison.
•The peristalsis can be cv<>; •<•.!. by ph. ini: .1 !•<•!! j.ir ..r n-llr' <-r ihc
'
i-iof. I. S.iMiatuni makes a window with .1 w.itdi class (Bioph. Ccntr., 4, 551, IQOQ).
mimal may be immersed in bath of warmed saline.
ii
162 A LABORATORY GUIDE IN PHARMACOLOGY
Paint the nicotin on the ganglion. Stimulation of the nerve is now ineffect-
ive, showing paralysis of the ganglion.
Experiment 8. Pilocarpin. — Expose the intestines freely. Inject intra-
venously 3 mg. per kg. of pilocarpin (3 c.c. per kg. of -fa per cent.) : the
peristalsis is increased (stimulation of the ganglia and muscle). Salivation
may be noticed (stimulation of salivary ganglia and endings). The heart
is at first slowed, but may be quickened later (peripheral stimulation and
depression of vagus).
The heart rate may be demonstrated by a. long needle piercing the heart
through the chest.
Experiment 9. Pituitary. — Inject intravenously pituitary solution, 0.5
c.c. per kg.: further increase of peristalsis.
Experiment 10. Atropin. — (a) Expose the vagus and determine the
smallest stimulus which will just stop the heart. Inject intravenously i
mg. per kg. of atropin (i c.c. per kg. of o.i per cent.): the peristalsis and
salivation cease (paralysis of endings). The heart is quickened, and stimu-
lation of the vagus becomes ineffective (paralysis of vagus endings). The
blood-pressure is not much altered; there may be a slight rise. (The rate of
the heart will not be changed by the atropin if the pilocarpin paralysis was
complete.)
Experiment n. Barium. — Inject intravenously barium chlorid, 10 mg.
(i c.c. of i per cent.) per kg.: strong peristalsis, even after the atropin.
Experiment 12. (Optional) Lead. — Anesthetized cat or rabbit, with window in ab-
domen. Inject into vein lead acetate, 5 to 8 mg. per kg.: intense peristalsis within live
minutes. Lumen nearly obliterated; vessels constricted. The spasm is relieved by intra-
venous injection of nicotin, atropin, or nitrites (Hirschfelder, 1915, Jour. Amer. }k<l.
Assoc., 65, 516).
QUESTIONS
(a) Describe the effect of stimulating the intestine by pinching.
(b) Would this reflex be useful for the propulsion of the contents? Why?
(c) What is the effect of physostigmin?
(d) Of barium?
(e) Of atropin?
(/) Of pilocarpin?
(g) Of pituitary?
(h) Which of the peristaltic stimulants are neutralized by atropin?
(i) How is it shown that nicotin paralyzes ganglia?
EXERCISE II.— (OPTIONAL) OBSERVATION OF PERISTALSIS ON
UNOPERATED RABBIT
Clip the hair from the abdomen of a rabbit which has been well fed two hours before.
Observe the normal peristalsis through the intact abdominal walls. Note the effects of
a sudden noise; of ammonia inhalation; of strongly pinching the skin over abdomen; of
hypodermic administration of nicotin, 10 mg. per kg.; then of atropin, 5 mg. per kg.
(J. Auer, 1907, Proc. Soc. Exp. Biol. Med., 5, 30).
EXERCISE III, A.— (OPTIONAL) ACID ON PYLORIC SPHINCTER
Remove stomach from twenty-four-hour fasting animal; place in warm oxygenated
Ringer's solution. Tie cannula in cardia and introduce small quantity of 0.4 c.c. HC1
with Congo-red (holding pylorus upward so it will not be touched by acid). Blow into
cannula tube until air bubbles through pylorus. Close cannula. When air ceases to
escape (i. e., when pylorus is closed), turn stomach gently so acid touches pylorus: this
open at once, so that blue fluid gushes out into the Ringer's solution. (Adapted from
Cannon, Movements, 106.)
CHAP. XXXIV SMOOTH MUSCLE! INTESTINE, UTERUS, AND ARTERIES
ent
log;
EXERCISE III, B.— (OPTIONAL) ACIDS AND ALKALIES ON TONE OF
CARDIAL SPHINCTER OF STOMACH
(See Cannon, "The Mci haniial Factors of Digestion," p. 40.)
EXERCISE IV.— (OPTIONAL) BAYLISS-STARLING REFLEX ON EXCISED
INTESTINE
Attach a piece of intestine, at each end, to water manometers. Fill with water and
suspend in a bath of warm oxygenated Tyrode fluid. On pinching the intestine the
manometer at the ascending end should show a temporary fall, the descending end a rise.
EXERCISE V.— (OPTIONAL) EFFECTS OF SMOKING ON HUNGER
CONTRACTIONS, HUMAN
See Carlson and Lewis, igi4, . \rner. Jour. Physiol., 34, 149.
EXERCISE VI.— (ALL GROUPS) AUTONOMIC POISONS ON RABBIT'S
INTESTINE
(REPORTER III, F)
Apparatus for Experiments on Excised Smooth Muscle of Mammals
(Intestines, Uterus, Bladder, Arterial Rings). — For each group arrange a
la rue water-bath maintained at 38° to 40° C. In this place a cylinder about
m. high and 3 cm. wide filled with 200 c.c. of warm Tyrode's solution.
One or two extra cylinders for changing the solutions may be kept in the
bath.
Arrange a muscle lever (Fig. 15) so —
that tracings may be taken from the in-
ine, etc., immersed in the solution.
When the tissue is in the cylinder a con-
tinuous stream of air or oxygen must be
bubbled through the solution. The stock
of tissues is kept in Tyrode's fluid, to
which the blood of the animal is added.
For periods longer than an hour the tissues
should be preserved in cold Ringer's fluid
in an ice-chest.
Technical
Smooth M
References. — Experiments
— Robert. lnt«>\.. i. 170.
on
3-
Fig. 15. — Arrangement for excised in-
! ".Mills,
1911, Tk't-r-trdt, 2.2. 141: Strwart ..}.}<'; NYukin h.
1912, Anh. m-s. IMi ('.nun and
Underbill. '
275; 'I o, Anh. Ink-mat, I'harmacod.,
2O, 205; M o.j. Anh. m>. I'hy>i<il.. 102,
132 (dilTt i
Tyrode's Solution.- I < r 1000: NaC'l, 8.0; KCI.
o.i; s i.o: ^lii. MM-, i
Localization of Action of Poisons in Intestines. M i MI ; i I
702,349; 108, i, 1005; (iunn and t'ndcrhill. loi.j. <,)u. irt. |<>m '
' rclcr. — M
Frog's EsoplMRut. — Stili-s. 1001. Ann r | .ur. l'h\ MO!., 5, 338; \\.iddcll. IQIO. il»id .
41,529-
Operation. Kill a rai -nale if the uterus experiment Unl>«-
made) by th« m artilu ial u-piiation. rapidly open the
1 64 A LABORATORY GUIDE IN PHARMACOLOGY
abdomen, insert a cannula into the abdominal aorta, and bleed dry. An
assistant will defibrinate the blood and add it to a liter of cold Tyrode's
fluid. (Excised organs preserve their excitability better when kept in the
cold; it may be for several days if laid on ice. Their contractions cease in
the cold, but resume on heating to body temperature.)
Excise the intestines in mass, also the uterus, and place in the Tyrode
blood mixture and pass current of oxygen and air.
From portions of the intestine which show active vermicular move-
ments cut pieces about 5 cm. long, attach to lever, immerse completely
in warm Tyrode solution, start air to bubbling, and take a slow tracing.
When a normal tracing has been obtained, and while lever is tracing on the
drum, add the drugs named below. The quantities are calculated for 200
c.c. of solution. If no response is obtained within a few minutes, further
doses of the drug may be added.
(Group I) Epinephrin, i drop of i : 10,000— inhibition; then Pilocarpin
in i c.c. of i : 1000 — contraction; then Atropin, i c.c. of i : 1000 — inhibition;
then Barium Chlorid, 2 c.c. of 10 per cent. — contraction.
(Group II) Pituitary Extract, 5 drops — stimulation; then Atropin, i c.c.
of i per cent. — no effect.
(Group III) Pilocarpin, i c.c. of i per cent. — stimulation; then Atropin,
i c.c. of i per cent. — inhibition.
(Group IV) Nicotin, i c.c. of i per cent. — stimulation; then Atropin,
i c.c. of i per cent. — inhibition.
(Group V) Barium Chlorid, 5 c.c. of i per cent. — stimulation; then
Atropin, i c.c. of i per cent. — no inhibition.
QUESTIONS
(a) Name the drugs which are stimulant and those which are depressant.
(b) Describe any differences in the character of the movements.
(c) Which of the stimulants act more peripheral than atropin?
(d) Assuming that atropin acts on the myoneural junction, state on which
structures the drugs named in (c) probably act.
(e) Which drugs act more central than atropin?
EXERCISE VII.— SALT ACTIONS ON INTESTINE
(REPORTER IV, F)
Fill the cylinder with warm 0.9 NaCl and immerse fresh piece of intes-
tine, arranged for tracing. Pass air; when slow normal tracing has been
taken, draw off the solution and replace by the following1 (previously warmed
in the bath):
(Group I) Sodium Sulphate, 1.9 per cent.2
(Group II) Sodium Citrate, 2.7 per cent.2
(Group III) Magnesium Chlorid, 2.1 per cent.2
(Group IV) Calcium Chlorid, 0.15 per cent.3 in 0.9 per cent. NaCl.
(Group V) Sodium Chlorid, 2 per cent.
(Optional) Water; Urea, 1.9 per cent.2; Cane-sugar, 10 per cent.2;
Sodium Phosphate, 2.1 per cent.2; Sod. Acid Phosphate, 2 per cent.2
i The percentages refer to the anhydrous salts.
These solutions have the same freezing-point as 0.9 per cent. NaCl.
* This corresponds to one-tenth of the isotonic quantity of the salt.
CHAP. XXXIV SMOOTH MUSCLE: INTESTINE, UTERUS,
QUESTIONS
(a) Describe the effects produced by the solutions.
(b) Which increase the contractions?
(c) Which diminish the contractions?
(d) Which increase the tone?
(e) Which relax it?
How is the effect of 2 per cent. NaCl explained?
•"»™»* 'OS. ogy
University
Toronto
j) Does the same explanation hold for the others? Why?
(h) Sodium sulphate and citrate, as well as magnesium chlorid, are used
as cathartics; calcium against diarrhea. Does this agree with their effects
on excised intestine?
EXERCISE VIII.— AUTONOMIC DRUGS ON UTERUS
(REPORTER V, F)
Explanatory. — The effects of drugs on the uterus are particularly im-
portant in obstetrics and toxicology. The effects differ according to species,
pregnancy, etc., but are essentially similar in intact animals and in excised
organs. The uterus is stimulated by ergo toxin, histamin, pituitary, etc.
Epinephrin contracts the uterus of the rabbit, dog, monkey and human, and
pregnant cat; it relaxes that of the guinea-pig and rat, and of the non-preg-
nant cat.
Experiment. — Cut pieces about 2 cm. long from cornu, arrange on
lever, immerse in warm Tyrode fluid; pass air, and take normal tracing.
Add the following drugs (per 200 c.c. of Tyrode fluid). If no response is
obtained within a few minutes, further doses may be added.
(Group I) Epinephrin, i drop of i : 10,000.
(Group II) Pituitary Extract, 5 drops.
(Group III) Quinin Hydrochlorid, 2 c.c. of o.i per cent.
(Group IV) Fid. Ext. Ergot, i c.c. (or 3 mg. of Ergotoxin).
(Group V) Tr. Hydrastis, i c.c.
TECHNICAL REFERENCES
Uterus in Situ. — Edmunds and Hale, see Exercise VII. Biagi (Centr. Bioch., 4, 762,
1905); Trendelenburg (Zs. Biol., 61, 67, 1913); Ruebsamen (clinical; Miuiuh. mid.
Woch., 2724, 1913); I'ittrnu'iT. 71; Harbour, 1915 (Jour. Pharmacol. l.xp. I lu-r . 7. 547).
Uterus, Excised. — Kurdinowski (Arch. Physiol., Suppl., 372, 1904); Kelm-r
Path. I'harm., 58, 366); Prochnow (Arch, ink-mat. Pliarmacod., 21, 305, 1911);
Pittenger, 73; Ounn (human); Proc. Roy. Soc., 87, 551.
General. — Robert, Intox., i, 257, -?KI; I'itk-ngrr, 91.
(Optional) Otlu-r drugs \\liit h may be used (Krlmr, 1007, Lieb, 1914) are (per 20.
1. Atropin, o.i, i. 10, and 500 mg.
2. Barium Chlorid, 60 mg.
3. Cotnrnin. X nig.
.\. //M/iJW/M. O
7w/rii.\//M. .;
6. 1/ytirtistinin. X mg.
' -rf>liin, 3.3 mg.
8. Nicotin, 10 t<> ;o mg.
I'Hof'irpin, 20 to 50 mg.
10. /'Ay.v.' ii» .'o mg.
i i. Slrnplitinthin. o..} to i mg.
12. 7 $ mg.
QUESTIONS
(a) State which of these drugs are depressant, and whirh stimulant.
Do the effects of epinephrin and pituitary agree with those on the
'•S?
1 66 A LABORATORY GUIDE IN PHARMACOLOGY
EXERCISE IX.— ARTERIAL RINGS (O. B. MEYER METHOD)
(REPORTER V, F)
Sheep's carotid artery may be obtained from the slaughter-house (it
can be kept in Ringer's fluid on ice for several days if necessary). It is
cut into rings about 2 mm. wide. One of these is suspended on a "heart-
lever" by thread passed through its lumen, so as to record the contraction
of the circular muscles. It is ^ immersed in warm Tyrode fluid and the
air-current started. The lever is pressed down several times (to overcome
the tonus) until it returns to a constant level. It is then made to trace
the base line on the drum. The drugs may be added as follows (per 200 c.c.) .
If no effect is obtained in a few minutes, the dosage may be increased:
(Group I) Epinephrin, i drop of i : 10,000; then Sod. Nitrite, i c.c. of
10 per cent.
(Group II) Sod. Nitrite, 10 per cent., drop by drop; then Epinephrin,
i drop of i : 10,000.
(Group III) Barium Chlorid, 5 c.c. of i per cent.; then Sod. Nitrite, i c.c.
of 10 per cent.
(Group IV) Tr. Digitalis, i c.c.; then Sod. Nitrite, i c.c. of 10 per cent.
(Group V) Physostigmin, i c.c. of i per cent. ; then Sod. Nitrite, i c.c.
of 10 per cent.
(Optional) Effect of Calcium on Excitability. — Cow, 191 1, Jour. Physiol., 42, 125.
(Optional) Antagonism. — Subject the rings to ergotoxin, i : 50,000: moderate but
lasting constriction. Change to epinephrin, i : 10,000: no effect or slight dilation (Macht,
1915, Jour. Pharm. Exp. Ther., 6, 591).
QUESTIONS
(a) Which of these drugs produce contraction and which relaxation?
(b) How does this agree with the effects on the intestines and uterus?
(c) Does the smooth muscle of different organs necessarily react alike to
a given drug?
TECHNICAL REFERENCES •
Excised Arteries. — O. B. Meyer, 1906, Zs. Biol., 48, 352; Stewart, 66; Cow, 1911,
Jour. Physiol., 42, 125; Barbour, 1912, Arch. exp. Path., 68, 41; Macht, 1915.
Elasticity of Arteries. — Tigerstedt, 2.4, 208.
OTHER SMOOTH MUSCLE
Ureter. — Lucas, 1906, Amer. Jour. Physiol., 17, 392; 1908, ibid., 22, 245.
Bladder. — Stewart, C. C., 1900, ibid., 4, 185; Trendelenburg (intact animals), 1913,
Zs. Biol., 61, 67.
Gall-bladder.— Lieb and McWhorter, 1915, Jour. Pharm. Exp. Ther., 7, 83.
Male Genitalia. — Robert, Intox., i, 216.
Invertebrates. — Tigerstedt, 1.2, 69; Robert, Intox., j, 154, 166.
EXERCISE X.— (OPTIONAL) USE OF SMOOTH MUSCLE IN BIO-ASSAY
The excised uterus and intestines are well adapted for qualitative and quantitative
tests, of which the following are important examples:
Experiment i. Ergot Test on Uterus in Situ. — Method of Edmunds and Hale. — A
non-pregnant cat is anesthetized with chloretone, 0.3 to 0.4 gm. per kg. by stomach-tube.
Cannulae are placed in jugular vein and trachea. Artificial respiration is started. The
animal is submerged in a bath of normal saline of 39° C. The uterus is exposed freely
through the linea alba. One horn is freed from its attachments and from the ovary.
Two threads are passed by a needle through the uterus, about 2 cm. apart. These
are fastened to a myocardiograph with light lever, put under proper tension, and tracings
taken, the drugs being injected into the vein. Injections are repeated every five to ten
minutes until same results are obtained as with the standard preparation; 0.2 to 0.3 c.c.
CHAP. XXXV REACTIONS OF BLOOD-VESSELS 167
of the Fluidextract of Ergot should cause distinct contractions. The method is tedious
and rather uncertain, especially if the uterus is making spontaneous contractions (Ed-
munds and Roth, 1008).
Experiment 2. Bio-assay of Pituitary. — The excised uterus is now generally used,
according to the method of Roth, igii. Jour. I'harmacol., 5, 559; Hyg. Bui. No. 100;.
U. S. P. IX. ( Uhrr u>t- are those of Dale and Laidlaw, 1912 (uterus), Jour. Pharmacol.,
Hamilton. 1012 (blood-pressure), Jour. Amer. Pharm. Assoc., i, 1117 (Pittenger, 88).
(Experiments on Pituitary, Robert. Intox., i, 267; Operations, Tigerstedt, 2.4, 98; on Pineal,
ibid., loo.)
Experiment 3. Bio-assay of Epinephrin. I.pinephrin may be tested in several ways,
and when its identity is to be established (for instam e. in serum) the simultaneous use of
several of these methods is indispensable, especially the intestine and uterus (G. N. Stewart.
ioi i . Jour. K.xp. Med.. 14, 377). For the quantitative comparison of commercial prepara-
tions or gland extracts the pressor effect on mammals (Chapter XLIII) is most convenient;
the j)erfusion of frog K-i:> I'hapu r \\.\\ and the arterial ring method (this chapter)
are also used. The mydriatic test (Chapter XXXVII) is employed for special problems.
The referenrei to these tests are summarized for convenience:
•on. — Crawford, 1907, U. S. Agr. Plant Ind., Bui. No. 112.
Mammalian Klood-pressure.—U. S. P. IX; Pittenger, 52; Elliott, 1912, Jour. Physiol.,
44, 374-
Presence in Blood. — Stewart, loc. cit.; Abdcrhalden's Handb., 6, 585.
Frog /Vr/ks/on.— Fuehner, 140; Trendelenburg, 1910, Arch. exp. Path., 6, 165; 1915,
ibid., 79, 154; Tatum, 1912, Jour. Pharmacol., 4, 151.
I'crfusion. — Swetschnikow, 1914, Arch. ges. Physiol., 157, 471.
Intestinal Method. — Cannon and La Paz, 1911, Amer. Jour. Physiol., 28, 64; Hoskins,
1911, Jour. Pharmacol., 3, 93.
.Stewart, loc. cit.
Pupil.— Abderhalden, 5, 112; Meltzer, 1909, Deut. med. Woch.. Xo. 13; Ehrmann,
1005, Arch. exp. Path., 53, 97.
Experiment 4. — Bio-assay of Charcoal Absorption. — Tracings are taken in the usual
manner from excised intestine. To 100 c.c. of Ringer's solution add o.i c.c. of hi--
tamin solution, i : 100,000: strong contraction. To another piece, suspended in fresh
Ringer's, add up to 10 c.c. of a histamin solution, of the same strength, but which has pre-
viously been shaken with blood charcoal, 3 gm. per 100 c.c. of histamin solution. Thi-
treated solution, filtered or unfiltered, should be ineffective (Guggenheim, 1915, Ther.
Monatsch., 29, 615).
CHAPTER XXXV
REACTIONS OF BLOOD-VESSELS (PERFUSION EXPERIMENTS,
ETC.)
This subject will be studied in further detail in connection with the
blood-preSSUre experiments. However, the peripheral effet t- may be shown
by pcrfusion experiment-, and -nine of tin- ireneral phenomena can be ob-
< d <>n intact animals. The behavior of excised arterial rin^s was noted
in the la.-t chapter.
Technical Notes on Perfusion. IVrfusion, especially of excised organs,
•••I to study the direct ctYcct- of drills upon their \< produce
artificial chai in ulat i..n ; to study their work under determinable
conditio;
me! hod OODSJ liallv in circulating liquid through \'
.111 under suitable condit ion- of pieSSUTe. 'I'he detail-
. ial object and accurdiii.u to the delicacy of the ti— ue. When
ing with a delicate function it i- necessary t mute precautions
as to the composition, oxygenation. temperature, and pressm ably
1 T»u A ;,s may alternate Chapters XXXV and X uccessive days.
i68
A LABORATORY GUIDE IN PHARMACOLOGY
pulsating) of the perfusion fluid. When investigating the more resistant
vascular reactions these complications are superfluous. It suffices to con-
nect the artery of the organ through a cannula with a reservoir of saline
solution, placed at a height approximating the normal blood-pressure (Fig.
1 6). Changes in the caliber of the vessels are denoted by corresponding
changes in the vein-flow from the organ. The organ — for instance, the kid-
ney— may also be placed in the oncometer (Sollmann and Hatcher, 1905,
Amer. Jour. Physiol., 13, 241).
The technkfor more elaborate perfusion is discussed by Franz Mueller, 1910, in Abder-
halden's Handb., 3, 321; 351; Tigerstedt, 1.4, 51; Kobert, Intox., i, 171; Friedmann,
1910, Zbl. Bioch. Bioph., 10, 864; Richards and Drinker, 1915,
Jour. Pharm. Exp. Ther., 7, 467.
Perfusion for Metabolism. — Abderhalden, 5, 1245.
Perfusion Reservoirs. — "Mercury bulbs" or "aspirator
bottles" of 200- to 2000-c.c. capacity may be used. To main-
tain a constant pressure the upper opening is furnished with
a "Mariotte stopper," /. e., a perforated stopper bearing a
glass tube which tips to near the bottom of the reservoir.
Constant Pressure. — This is obtained most conveniently
by raising the reservoir to the desired level — usually i to i£
meters above the organ — joining it to the arterial cannula
by alternate sections of narrow rubber and glass tubing and
closed by a pinch-cock. A T-tube, inserted just before the
arterial cannula, is convenient for removal of air-bubbles,
which must never be allowed to enter the vessels. The T
also serves for connection with a second reservoir if the solu-
tions are to be changed.
Warm Perfusion. — A Woulfe bottle filled with the solution
and immersed in a water-bath is interposed between the reser-
voir and the organ. The tube coming from the reservoir
tips to the bottom of the bottle; that going to the organ tips
about one-third down. The third tubulure bears the thermom-
eter. The organs are supported by cotton, or laid in a bath
of warm oil, or suspended in a hot-water funnel (such as is
used for filtering gelatin). This allows good drainage.
Rhythmic Pressure. — Ths is obtained by rhythmically
compressing the delivery tube or by opening a side tube (for
instance, Gesell, 1914, Amer. Jour. Physiol., 34, 186; for frog,
Verworn, Erregung and Laehmung, 164).
Oxygen Pressure. — If the solution is to be oxygenated,
the oxygen may be used to furnish the pressure, regulating
this by a mercury valve (for instance, in the Langendorff
heart perfusion apparatus).
Perfusion Stop-cocks. — When a series of fluids are to be alternated several-way stop-
cocks may be convenient. They are described by Locke, 1908, Quart. Jour. Exp. Physiol.,
i, 370; Macmillan, 1911, Jour. Physiol. Proc., July 22; Mines, 1913, Jour. Physiol., 46, 190.
Measurement of Vein-flow. — The perfusion-flow is estimated most conveniently by
the quantity of fluid flowing from the vein. If the changes are relatively slow, it suffices
to insert an elbow cannula into the vein and collect the fluid, determining either the
quantity collected in a given time or the time required to collect a given volume.
If the changes are fairly rapid, the flow may be measured by a dipping bucket (W. R.
Williams, 1910, Jour. Pharmacol., i, 457; Condon, 1913, Proc. Physiol. Soc., Jour. Physiol.,
46); or a Ludwig slromuhr (Sollmann and Pilcher, 1910, Amer. Jour. Physiol., 26, 236).
Other methods, used especially for vein-flow in intact animals, are those of Barcroft and
Brodie, 1905, Jour. Physiol., 33, 53 (rise of tambour); Wiggers, 1908, Amer. Jour. Physiol.,
23> 23 (scale pan); Brodie and Vogt, 1910, Jour. Physiol., 40, 135 (oncometer); Brodie
and Russel, 1905, Jour. Physiol., 32; Ishikawa and Starling, Jour. Physiol., 45, 164;
Burton-Opitz, 1908, Arch. ges. Physiol., 121, 150 (vein stromunr); W. Trendelenburg,
1914, Zs. Biol., 65, 13; see also Tigerstedt, 2.4, 259; Heinz, 2, 145; Kobert, Intox., i, 233.
Drop Recorders. — These are used when the flow of liquid is slow. A simple type
is shown in Fig. 17. In demonstrations a small electric lamp may be inserted in the
circuit. Another simple type is described by Fuehner, Nachweiss, p. 143. See alsa
Abderhalden's Handb., 5, 109; Macmillan, 1913, Quart. Jour. Exp. Physiol., 6, 109.
Fig. 1 6. — Diagram of kidney
perfusion.
CHAP. XXXV
REACTIONS OF BLOOD-VESSELS
I69
Oncometers Plethysmographs). — These are instruments for observing and measur-
ing changes in the volume of an organ. A very >imple form may be made of a conve-
niently shaped tin box, which has two openings, one for the vessels of the organ, another
for the tube of the recording apparatus. This consists of an elongated thin rubber bag
(such as is used in toy balloons), connected with a water manometer. The bag is filled
with water, connected with the manometer, and folded about the organ within the box.
\Yhen the latter is closed, any change in the volume of the organ is communicated through
the bag to the manometer. It may be recorded by connecting the free limb of the man-
ometer with a Brodie bellows or piston-recorder.
More elaborate forms are described by Roy, Schaefer's Textbook, i, 643; Schaefer
and Moore, 1896, Jour. Physiol., 20, i (gutta percha); Edmunds, Jour. Physiol., 22, 380
(intestines, plaster); 1913, Zs. Immun.. 17, 119; Jour. Pharmacol., 5, 520, 1914; Jour.
Pharmacol., 6, 589, 1915 (liver); Cloetta, 1010, Arch. exp. Path., 63, 147 (lung); also
Tigerstedt, 2.4, 272; Heinz, 2, 154. Plethysmograph for extremities and Recording
Devices , see Exercise \ III.
Preparation of the Organs for Perfusion. — The animal is usually bled. (If the per-
fu>ion is to be made with diluted blood, a liter or two of Locke's solution is run into the
femoral vein and the animal is again bled.) The bloods are defibrinated by whipping,
strained through cloth, and poured into the reservoir. The organ is exposed, a cannulu
: 7. — Drop marker: A small mica slide (m) is fixed at the end of the muscle-lever by means
nail cork. The mica slopes downward. The lever is kept horizontal by a long band of thin
elastic rubber (e), so that a drop falling on m will cause the pin p to dip into the mercury in the hollow
cork c. dosing the circuit with the battery b, and moving the magnet s, which writes on the drum.
The outflow tube is placed at least a foot above the mica slide.
is tied in it- artery, and connected with the reservoir. The vessels are well flushed (to
prevent dotting). The vein cannula is now tied in. All other vessels are tied and tin-
organ is removed. To avoid drying it may be covered with a IIHIM le skin tlap from the
abdomen of the dead animal.
Cannulee. A plentiful assortment of different -i/r- and form- -hould be on hand.
They are best made from glass tubing. The edges should not be sharp; they may be
rounded in the flame or on a sandsi.
Vessel Ctnnulae.— Fig i*. <i to ,1. shows the shape and the most useful sizes: a is
e in the frog's heart; /• for rabbit's .arotid or d«-g'> femoral artcr\ ; < for dog's carotid
.\tcrnal jugular. A still smaller size is needed for
glandular d
These (annula- are made by heating the prop< a large blow pipe
and drawing it out in the for: I his i- allowed to < <>o| and < ut at a.
The piet e- are then heated with a very -mall pointed flame at t. SO as to make the shoulder.
;t olT as obliquely U pOttiblc by M rat. lung with a triangular hie. ground
to i! and rounded in the llamc \ | I .annula should
have the e: ntly large so that it will not slip when tied into the vessels, but no
larger.
at ing glass, it should be constantly rotated in the flame; it is well to push it
170
A LABORATORY GUIDE IN PHARMACOLOGY
together very gently while heating. It should always be removed from the flame before
drawing.)
Tracheal Cannulae. — These are of the form shown in Fig. 18, <•.
One end is best made somewhat smaller than the other, so that the same cannula
may serve for somewhat different sizes of trachea. Tubing 5 and 8 (Fig. 20) is most use-
ful for rabbits; 9, 10, and 12, for dogs. The Harvard metal cannulae serve excellently.
V v
Fig. 1 8. — Cannulae for vessels and trachea.
Aortic and Bladder Cannula. — This is made of the form and size of Fig. 21. The
rings are made by heating a narrovy zone of the tube in a small flame, and pushing the
glass together. When used on the bladder, this cannula is tied in the neck. Another
bladder cannula, used especially in rabbits, consists of a short thistle tube (Fig. 22).
The bladder is cut open and tied as a drum-membrane over the mouth of the cannulae,
the ureters being left free and opening into the cannula.
Fig. 19. — Tube drawn for cannulae or pi pets.
Ureter cannulae are given the form shown in Fig. 23. This is the proper size for dogs.
A smaller tube is required for rabbits. The narrow tubing is obtained by using the portion
between the arrows in Fig. 19, making this somewhat longer.
Insertion of Cannulae Into Vessels. — The vessel is exposed and cleared of all fascia
for the space of i inch, if possible. A bulldog forceps1 (Fig. 24) is then applied to the end
of the vessel toward which the cannula will point. A ligature is passed by forceps or
aneurysm needle around the vessel near the clamp, and tied into a loose slip-knot. The
vessel is then allowed to fill with blood, and another ligature tied securely as far away
OOO0
Fig. 20. — Sizes of glass tubing.
irom the clamp as possible. The vessel is now lifted by the end of the second ligature
and laid on the left index-finger. An incision is made with small curved scissors near the
distal ligature, about two- thirds through the vessel, the moistened point of the cannula
is pushed in, and the loose ligature is tied securely around the neck. The ends of the
ligatures are now cut off. The largest cannula should be chosen which will fit the vessel
without force. The cannula is turned within the vessel so that kinking will not close
the opening of the cannula.
1 When buying these clamps one should take care that the jaws touch along their entire surface.
CHAP. XXXV
REACTIONS OF BLOOD-VESSELS
171
The whole procedure is quite easy when the vessels are strong. Delicate vessels
should be well distended, and all twisting must be avoided. It may be necessary to hold
the vessel open with very fine-pointed forceps. The manipulations must be made \ cry
delicately.
Ligatures. — It is a mistake to use ligatures which are too thick. The following are
useful sizes: No. 50 linen thread or buttonhole-twist silk for vessels; cotton wrapping
twine for trachea, bladder, etc. They should be cut to a length of about 6 inches. (This
may be done in mass by winding the string around the palm of the hand.)
Fig. 2 1 . — Aortic and bladder cannula. Actual size. Fig. 2 2 . — Bladder cannula.
A ligature should be tied as securely as its strength will allow. A little practice will
show its limitations. More force can be exerted if the pull is made very near to the knot.
A plain double knot is best for small vessels; the bulky surgeon's knot should be confined
t«» larger structures, such as the trachea or aorta.
Ureter cannulse are introduced in the same manner as described for the vessels, except
that the ureter need not be clamped.
The same general method is also used for inserting the tracheal cannula. The trachea
is exposed, cleaned, two ligatures are placed i or 2 inches apart, and three or four rings of
cartilage are divided with the knife by a straight or V-shaped incision.
Fig. 23. — Ureter cannula. Actual size.
Perfusion of Brain. — Dixon and Halliburton, 1910, Quart. Jour. Exp. Physiol., 3, 315;
in situ, E. D. Brown, 1916, Jour. Pharmacol., 8, 185.
Perfusion of Liver.- - Ba^lioni. Abderhalden, 3, 364; Macleod and Pearce, 1914, Ann r.
Jour. IMiy-iol.. ,^5, 87; Frog, Morita, 1915, Arch. exp. Path. Pharm., 78, 232.
Perfusion of Lung.— Hachr and Pick, 1913, Arch. Exp. Path., 74, 42; Tigerstedt. ..4,
296; Magnus and S« 1914, Anh. ges. Physiol., 155, 192; Modrakowski, ibid.,
158, 509.
Fig. 24. — Bulldog clamps.
Coronary Perfusion. — Morawitz and /aim \r«h. Klin " . 364.
Heart-lung-kidney Preparation.— Hainbridm- and I \.rn-. 1014. Jour I
278.
Splanchnic Vessels, Frog.— Frochlirh and Morita. 1015. An h. rxp. Path. IMiarm., 78,
' Perfusion Fluids.- Tin- plain .V«»rw,;/ S.ilinc .SW;«
r.-nd. h the Mood MTUMI
~j). They suffice for injY. ti«.n> into li\ii^ animals, hut n- i-d tissues or
172
A LABORATORY GUIDE IN PHARMACOLOGY
perfusions. For these it is necessary to use more complex fluids. The more important
of these are shown in the following table (also Tigerstedt, 2.4, 170).
*
TABLE OF COMMONLY USED BALANCED SOLUTIONS
Author.
^
Adapted to —
PERCENTAGES:
NaCl.
KC1.
CaCh.
NaHCOa.
Other ingredients.
Ringer.
Frog's heart.
0.6
0.0075
o.oi (dried).
O.OI
0.026 (crystals).
Howefl.
Frog's heart.
0.7
0.03
0.025 (crystals).
0.003
*
Clark.
Frog's heart.
0.7
0.014
o.oi 2 (dried).
O.O2
Goethlin.
Frog's heart.
0.65
O.OI
0.0065 (dried).
O.OI
/Na2HPO4, 0.0009
\NaH2PO4, 0.0008
Locke.
Mammalian
0.92
0.042
0.024 (crystals).
0.015
Dextrose, o.i
heart.
Rusch.
Mammalian
0.8
0.0075;
o.oi (dried).
O.OI
heart.
MgCl2, o.oi
Tyrode.
Mammalian
0.8
O.O2
0.02 (crystals).
O.I
Na2HPO4, 0.005
intestine.
m
Glucose, o.i
MgSO4, 0.03
Hedon and
Mammals.
0.6
0.03
o.oi (dried).
0.15
Na2HPO4, 0.05
Fleig.
Glucose, o.i
MgCb, 0.025
Adler.
Mammals.
o-59
O.O4
0.04 (crystals).
0.351
Na2HPO4, 0.0126
Glucose, 0.15
* The lower K content gives a more rapid heart-rate.
Note i. — In making solutions containing NaHCOs, this must be completely dissolved
before the CaCl2 is added.
Note 2. — Other solutions are described in "Digests of Comments on Pharmacopoeia,"
1911, p. 611.
Stock Solutions. — As the perfusion fluids are often used in considerable quantities
it is convenient to prepare them as concentrated stock solutions twenty times the original
strength. The concentrated calcium solutions should be kept separate, and added after
the other ingredients are diluted.
Solutions of Salts giving the same freezing-point as i per cent. Sodium Chlorid
(i gm. of NaCl added to 100 c.c. of distilled water; A = 0.589; molecular concentration =
0.316).
All the salts are to be weighed in grams and made up to i liter with distilled water.
They should first be dried to constant weight at 110° C. unless otherwise stated. They
must always be controlled by actual freezing-point determination.
Checked by the Author:
2 ................ 25.62
i ................ 16.33
Cl ...... 10 c.c. = 15.8 c.c.
n/10 NaOH
LiCl. .
MgCl2 .............
Na Acetate ........
NaHCO3 ..........
(Do not dry)
NaClO3 .............. 17.95
Na Citrate.
-33 to
NaNO., ............... 15.35
Na Oxalate ........... 23.00
Na2HPO4 ............. 21.00
7.26
21.15
12.75
9.66
(47-33 to 75.73 crystals)
27-37
Deduced from Published Tables:
Alcohol 14-50
plus i liter
Cane Sugar 108.82
plus i liter
Glucose 56.74
plus i liter
Urea 18.94
plus i liter
MgS04 35.37
Na2C03 14.54
NaOH 7.00
Deduced by A nalogy:
NH4C1 9.13
NaBr 17.46.
Nal 25.42
NaCNS 14.24
NaF 7.21
(47.73 crystals)
CHAP. XXXV REACTIONS OF BLOOD-VESSELS 173
EXERCISE I.— (DEMONSTRATION) NICOTIN ON EAR VESSELS. (VASO-
DILATION FROM DEPRESSION OF VASOCONSTRICTOR GANGLIA.
VASOCONSTRICTION THROUGH REFLEX STIMULATION.)
(REPORTER I, C)
Inject a white rabbit with 10 mg. per kg. of nicotin (i c.c. of i per cent,
per kg.): in about ten minutes the ear vessels are seen to dilate. (De-
pression of the sympathetic ganglia.) Apply reflex stimulation (blowing
on the rabbit): the vessels constrict at once; after a short time they dilate
. and the experiment may be repeated indefinitely. (The small dose
of nicotin used in this experiment produces a depression of the ganglia
sufficient to block the weak tonic vasoconstrictor impulses which pass
normally to the muscle: but it is not sufficient to block stronger impulses, as
those due to reflex stimulation. Larger doses of nicotin block these im-
pulses also.)
The $ IK nil c/ect of nicotin may also be observed on this animal. The
reflex excitability is first increased, then the animal shows a condition of
partial paralysis, with convulsions on stimulation. There may be nausea.
The pupils are variable.
Questions. — (a) What vasomotor changes are produced by nicotin?,
(b) Describe the symptoms of nicotin poisoning.
EXERCISE II.— (DEMONSTRATION) ERGOT ON COMB OF ROOSTER
(REPORTER I, C)
Administer to a rooster 5 gm. of powdered ergot (rolled into a cartridge
with tissue paper) by mouth, or 5 c.c. of fluidextract hypodermicallv.
\\ithin an hour the tips of the comb and wattles will become cool and
blacken. This may persist for several days and may pass into dry gangrene
of the affected parts. The result is due either to a persistent vasoconstric-
tion result ing from a direct action on the arterial muscle, or to some change
in the endothelium. (The experiment is often unsuccessful if the ergot has
become inactive, or if the animal is not very susceptible.)
EXERCISE III.— (OPTIONAL) ASSAY OF ERGOT ON ROOSTER-COMB
This is probably tin- mo>t reliable test for the activity of ergot. The official method
is described in the U. S. P. (also Pittenger, 69).
EXERCISE IV, A.— (DEMONSTRATION) PERFUSION OF FROG'S VESSELS
(LEWEN-TRENDELENBURG METHOD)
I. C)
The method CQDSifttfl in the perfusion of the legs of the pithed
through the abdominal aorta from a Mariotte bottle. The outflow from
the abdominal vein i- recorded by a drop-counter. The drug is injected
with a syringe into the tubing leading to the aorta. The flow i- |]0wed
by constrictor drugs, and vice versa. The details are as follows:
Decapitate a large frog and pith tin- >pinal ">r<i \ itrip < in. \\id«.
away from th« .ibd«>m<-n. The sternum
domin.il \tm U divided j 1 a strip of the abdominal wall with
l In- le^'s. iriiiilrs ai.'vdu-ntrs.
174
A LABORATORY GUIDE IN PHARMACOLOGY
the thigh toward the kidneys, are surrounded by ligatures and tied. The abdominal
organs are then removed, avoiding injury to the aorta or abdominal vein.
The frog is now fixed to a cork-board (Fig. 25). A very fine, long-pointed tannula.
is tied into the aorta, so that its point is just above the bifurcation. The cannula \va>
previously connected by rubber tubing (about 40 cm. long) with a 25o-cm. Mariotte bottle
(Tech. Notes), filled with Ringer's fluid. The connecting tube bears a screw-clamp,
which is opened slightly during the introduction. Air bubbles must be rigorously ex-
cluded. The tubing is fastened to the board. When the fluid drops from the abdominal
vein a thin glass tube, about i mm. in di-
ameter and about 6 cm. long, is tied into
the vein. The free end of the tube is bent
to facilitate dropping and raised somewhat
over the board by a small cork.
The Mariotte bottle (Fig. 26, MF) is
now adjusted at such a level (perhaps 1 5
cm. above the frog) that the vein delivers
30 to 40 drops per minute. A drop-counter
Fig. 17, p. 169, is arranged under the drops.
The marker is adjusted on a drum, to-
gether with a time-marker tracing second.
Fig. 25. — Frog preparation for vessel perfusion
(Fuehner): (a) Aorta; (b) abdominal vein; (e)
ligated rectum and bladder; (d) venae renales
advehentes.
Fig. 26. — Frog perfusion (Fuehner).
After a normal tracing has been taken, £ or i c.c. of the solution to be tested is in-
jected very slowly with a hypodermic syringe into the connecting tube. The injection
should raise the fluid in the glass tube of the Mariotte bottle by about i cm., and should
occupy about fifteen seconds. An injection of Ringer's fluid solution may first be madi-
as a blank test to discount the mechanical effects of injection. (Tatum, 1912, Jour.
Pharmacol., 4, 151, describes an arrangement for eliminating the disturbance.)
The sensitiveness of the vessels increases for several hours. The occurrence of edema
is not detrimental.
TECHNICAL REFERENCES
Trendelenburg, Deut. Arch. klin. Med., 103; Arch. exp. Path. Pharm., 63, 165;
ibid., 1915, 79, 154; Fuehner, Nachweiss, p. 140; Tatum, 1912, Jour. Pharm. Exp. Ther.,.
4, 151-
CHAP. XXXV
REACTIONS OF BLOOD-VESSELS
175
The following solutions may be tried (dissolved in Ringer's fluid):
Sod. Nitrite, i : 1000; then Epinephrin, i : 5,000,000; then Digitalis, i : 100.
(Optional) Synergism of Epinephrin and Serum. — Compare the following on the
Trendelenburg preparation:
(a) Ringer's solution.
(b) Ditto, injecting i c.c. of Serum, i : 4.
(c) Ringer's solution.
(d) Ditto, with addition of Epinephrin, i : 100,000,000.
1 1 '
1
(e) Ditto, ditto, injecting i c.c. of Serum, i : 4.
Rinner's solution.
) Ditto. injeiting i c.c. of Epinephrin, i : 10,000,000.
t) Ringer's solution.
>') Ditto, with addition of Serum, i : 150.
k) Ditto, ditto, injecting i c.c. of Epinephrin, i : 10,000,000.
(Moog, 1914, Arch. exp. Path. Pharm., 77, 346.)
QUESTIONS
(a) Describe the effects of llu-st- drugs.
(b) On what structures are the actions exerted?
EXERCISE IV, B.— (OPTIONAL) PERFUSION OF ISOLATED RABBIT'S
EAR
(Bissemski) Rischbieter, 1913, Zs. ges. exp. Med., i, 355; Swctsehnikow, 1914, Arch.
ysid., 157, 47i.
EXERCISE V.— (OPTIONAL) MICROSCOPIC OBSERVATION OF VESSELS
Experiment i. Digitalis on Vessels of Frog's Foot. — Curari/.e a frog. Pin on board
to observe circulation in foot (Oc. Ill, ohj. III). Make an exact drawing of a small
vessel. Inject into lymph-sac 0.5 C.C. of tincture < 10 per cent.) of digitalis and oh>erve the
vr—el from time to time and note change- in its diameter. A marked vasocon-
ibout 25 per cent.) is usually observed. Ergot, 0.5 c.c. of fluidextract, max
be used.
r\
Fig. 27.— < (>oard, for studying the rinul.ition in the frog's omcntum; I actual sire.
More exa< ' m be obtained 1 u < e mi* mm
For observing the circulation of lh<- frog's foot a triangular >lit i- « ut from om
the hoard, ;in<! ihr \\rh ..f the i 'his slit. 'I'his is lai<i «>n the stage of
the r: the otlu-r end of the hoard bring . .mvrnirntly s\ip|Mirtrd by a tunil
ire, the frog may be anesthetized by uretlianc (Oehr\\all
Skand. Anh. Phy>iol., j5, i).
£76 A LABORATORY GUIDE IN PHARMACOLOGY
Further descriptions of the method are found in Tigerstedt, 2.4, 312; Heinz, 2, 144;
Robert Intox., i, 195; Cohnheim, Virch. Arch., 40.
Experiment 2. Mesenteric Vessels. — For observing the circulation in the omctitum
the cork-board shown in Fig. 27 is employed. A semicircle is cut out at one side to adapt
it to the stand of the microscope. A hole of about 18 mm. is made near the center with
a cork-borer. Into tins a perforated cork (i cm. bore) is pushed tightly. The bottom
of the cork is cut off flush with the board. The top projects i cm. above the board. The
edges of the cork are rounded with a file.
To observe the circulation, the brain of the frog is pithed. The abdomen is opened
and the sciatic nerves divided within the abdomen. The frog is then pinned on the
board on the side away from the microscope, so that the abdomen touches the cork.
A small pledget of cotton, moistened with normal saline solution, is inserted between the
frog and the cork. A coU of intestine is drawn out carefully and pinned over the cork,
so that the mesentery comes to lie over the opening. Twisting of the vessels must be
avoided. A triangular piece of filter-paper is laid with its base on the opened abdomen
and its apex on the mesentery. This is moistened with normal saline solution.
Epinephrin, o.oi per cent., may be tried.
The experiment may be modified to show the action of astringents by first inducing
an inflammation and then applying i per cent. alum.
EXERCISE VI.— (OPTIONAL) BLOOD-PRESSURE OF FROGS
See Jacoby and Roemer, 1911, Arch. Exp. Path. Pharm., 66, 270; Burket, 1913,
Kansas Univ. Sci. Bui., 17, 219; Kuno, 1914, Arch. ges. Physiol., 158, i; Schulz, 1906,
Arch. ges. Physiol., 115, 386; Toads, Tigerstedt, 2.4, 211.
EXERCISE VH.— (ALL GROUPS) PERFUSION OF MAMMALIAN KIDNEYS
AND OTHER ORGANS
(REPORTER II, C)
Experiment i. (Group I) Mechanical Changes in Circulation. — (See
Technic, page 167.) Perfuse dog kidney with 2 per cent. NaCl, as described
above, observe vein and ureter flow (drops or cubic centimeters per minute)
and oncometer.
(a) Effect of Arterial Pressure. — Start with the reservoir at 140 cm. above
the kidney. Make observations after fluid has run for about ten minutes.
Lower the reservoir to 100 cm., and repeat the observations after ten min-
utes; also with 60 and 20 cm.
The vein flow, ureter flow, and oncometer (also the maximal vein and
ureter pressure) vary in the same direction as the arterial pressure.
(By modifying the arrangement so that the pressure can be interrupted
rhythmically, it can also be shown that the vein and ureter flow are much
better with interrupted pressure than with constant pressure of the same
mean height.)
(b) Effect of Vein Pressure. — Replace the reservoir at 140 cm. Remove
the outflow-tip from the vein cannula and connect this with a rubber tube
i m. long. Replace the outflow-tip in this tube and support it at the level
of the kidney. Let it fill with the fluid, and in ten minutes measure the
vein and ureter flow and the oncometer. Raise the vein outflow to 30 cm.
above the kidney and in ten minutes repeat the observation ; also at 60 and
90 cm. Increase of vein pressure increases the oncometer, but diminishes
the vein and ureter flow. The diminution is gradual up to 60 or 80 cm.,
when there is a sharp drop.
(c) Effect of Ureter Pressure. — Remove the tube from the vein and con-
nect it with the ureter cannula. Repeat the observations as in (b). The
effects are similar, but the ureter pressure has a comparatively small effect
on the vein flow and oncometer.
(d) Occlusion of the Vein. — Disconnect the tube. Count the ureter flow
Depart rneni
CHAP. XXXV REACTION'S OF BLOOD-VESSELS 177
and observe the oncometer. Pinch the vein tube to complete occlusion. ,
The oncometer increases. There is a short spurt of ureter fluid, and theft*
almost (but not quite) complete anuria (compression of the injury tubuie&nunto
the boundary layer).
(e) Injection by Renal Vein. — Release the vein and after ten minutes
count the vein and ureter flow and observe the oncometer. Change the
injection tube from the artery to the renal vein: almost no fluid will run
from the artery or ureter, the oncometer increasing greatly. (A valvular
mechanism exists in the kidney, probably by the pressure of the distended
veins on the arterial capillaries in the glomeruli.)
(Jut-xtions. — (a) Describe the effects of arterial pressure.
(b) At what level does the filtration of "urine" stop?
(c) Do these facts agree with what is observed in intact animals?
(d) Describe the effects of vein-pressure.
(e) Do they agree with those in intact animal-:'
(/) Why does increased pressure in the renal vein diminish the filtra-
tion of "urine"?
(g) Describe the effects of ureter pressure.
Is temporary anuria on compression of renal vein a valid argument
:ist the physical filtration of urine?
(/) Can the circulation in the kidneys be reversed? Why?
Experiment 2. (Group II) Salt Actions on Kidney. — Use two bulbs con-
nected with J-piece, one filled with i per cent. XaCl, the other with water.
(a) Perfuse the kidney with i per cent. NaCl solution, and observe the
vein and ureter flow (drops per minute) and the oncometer after ten minutes.
(b) Hypo-isotonic Solutions. — Replace the salt solution by water. The
vein and ureter flow and the volume are diminished. This is due to the
swelling of the renal cells obstructing the access of the fluid to the kidney.
(c) Hyper isotonic Solutions. — Replace by 5 per cent. XaCl: the flow
increases much above the original, the volume to about the original (less-
ened resistance by shrinkage of cells.)
Return to i per cent. XaCl solution. After fifteen minutes replace this
by:
< dlcium Chi or id (1.6 per cent, of anhydrous, isotonic with i per cent.
NaCl). — The flow and oncometer are diminished. This is a specific (ion)
effect of the calcium.
(e) Citrate.— Replace by isotonic sodium citrate (2.75 per cent, of anhy-
drous: the flow and oncometer are increased. The citrate acts as a
hyperisotonic solution, since it does not penetrate the cells a> readi
i (consult Exercise 23, No. 3).
(/) ' of I Y/;/.— Pinch the tube of the vein-cannula to complete
occlusions. (See Experiment i
Questions. — (a) Describe and explain the effects of hypotonic soli/
(b) Simii are produced in intact animals by the intrnvenoi;
of water; whereas the oral ingest ion i- diuretic. Kxplain the difference.
(c) Describe and explain the efl'ei t- of hypertonic solution-.
Doc- thi- explain that hypcridycemii animal- are nearly alwa\ >
polYi
(e) Describe the effei t Sum.
(/) ll»v. can tl Why?
Experiment 3. (Group III) Vascular Drugs on Perfused Kidney.
two hr.ll.- iMMiiected with J-piei e. one tilled with I per cent. XaCl, the
other with the drug di»olved in i per cent. NaCl.
178 A LABORATORY GUIDE IN PHARMACOLOGY
(a) Perfuse with i per cent. XaCl. After fifteen minutes observe the
vein and ureter flow (drops per minute) and the oncometer.
(b) Epinephrin. — Change to i : 50,000 (i c.c. of i : 1000 to 500.0. of i per
cent NaCl) : the flow and volume are diminished (constriction of arterioles).
(c) Hydrocyanic Acid. — Change to i : 2500 HCN (2 c.c. of 2 per cent, to
100 c.c. of i per cent. NaCl): increase of vein, ureter, and oncometer (dila-
tion of arterioles). This effect of hydrocyanic acid seems to be confined
to the kidney.
(d) Digitalis. — Change to i : 1000 Digitalis (i c.c. of 10 per cent, to 100
c.c. of i per cent. NaCl): vasoconstriction.
(e) Chloral. — Change to i : 1000 Chloral (i c.c. of 10 per cent, to 100 c.c.
of i per cent. NaCl) : vasodilation.
(/) Barium .—-Change to i : 2000 Barium Chlorid (5 c.c. of i per cent, to
100 c.c. of i per cent. NaCl): vasoconstriction.
(Questions. — (a] Which of the drugs produce vasoconstriction?
(b) Which produce vasodilation?
Experiment 4. (Group IV) Blood and Drugs on Excised Kidney. — Use
two bulbs connected with T-piece, one filled with i per cent. NaCl, the other
with the blood, etc.
(a) Perfuse with i per cent. NaCl. After fifteen minutes observe the
vein and ureter flow (drops per minute) and oncometer.
(b) Blood. — Dilute with about three volumes of i per cent. NaCl: the
vein flow is promptly increased, while the ureter flow and oncometer are
greatly diminished. Note the darkening of the venous blood. The flow
is again somewhat slowed after a time.
Two factors are concerned in these effects: The great viscidity of the
blood, which would slow the flow and diminish the volume. In dead
kidneys the vein flowr is practically arrested. In living kidneys, however,
the blood stimulates a vasodilator mechanism, probably in the efferent
arterioles, which causes the vein flow to continue, and generally increases it
above normal.
(c) Saline Diuretics. — After about fifteen minutes repeat the observa-
tions. (The vein flow will be somewhat slowed on account of the increasing
viscidity.) Add about 30 per cent, of i per cent. NaCl to the perfusing
blood: the flow and volume are increased (lessened viscidity, and conse-
quently lesser resistance). Use this blood dilution in all subsequent ex-
periments.
(d) Caffein. — Substitute diluted blood with i : 5000 Caffein (i c.c. of
i per cent, to 50 c.c. of diluted blood) : somewhat increased flow and vol-
ume. (Not always successful.)
(e) Hydrocyanic Acid. — Substitute blood w?th i : 2500 HCN (2 c.c. of 2
per cent, to 100 c.c. of diluted blood) : further vasodilation.
Note that the venous blood is not darkened, but that it is readily reduced
by ammonium sulphid. (Cyanids prevent the reduction of blood by
paralyzing the oxygen-consuming metabolism of the cells.)
(/) Digitalis. — Substitute blood with i : 1000 Digitalis (i c.c. of 10 per
cent, to 100 c.c. of diluted blood) : strong vasoconstriction. (More dilute
solutions cause some dilation.)
Questions. — (a) What effect has blood on the renal circulation?
(b) Describe the effects of adding saline solution.
(c) How do these compare with those of hydremia in intact animals?
(d) How does this explain increased diuresis when effusions are being
absorbed?
CHAP. XXXV REACTIONS OF BLOOD-VESSELS 179
(e) Describe the typical effects of caffein.
(/) Could the diuretic action of caffein be due to its action on the kidney
vessels?
(g) Describe the effects of hydrocyanic acid.
(h) What changes does it cause in the color of the blood?
(0 How are these explained?
(k) What effect would such an action have on an animal?
(/) Describe the effects of digitalis.
Experiment 5. i Group V) Circulation Through Excised Spleen or In-
testine.— In this exercise the drugs are injected slowly into the circulation
by means of a hypodermic syringe. The experiment may be modified by
adding the drugs directly to the perfusing fluid, and by using cold or warm
defibrinated blood.
Perfuse with i per cent. NaCl and in ten minutes observe the rate of flow
(drops per minute) and oncometer:
Suprarenal. — Inject 5 c.c. of i : 10,000 epinephrin: vasoconstriction.
.\itritcs. — Inject 5 c.c. of i : 100 sodium nitrite: vasodilation.
•'alls. — Inject 5 c.c. of i : 100 digitalis: vasoconstriction.
Chloral. — Inject 5 c.c. of i : 100 chloral: vasodilation.
Barium. — Inject 5 c.c. of i : 1000 barium chlorid: vasoconstriction.
Instead of injecting the stronger solutions, weaker concentrations may
be perfused as in Experiment 3 :
Epinephrin, i : 50,000 = 2 c.c. of o.i per cent, to 100 c.c. of i per cent.
XaCl.
Sodium Nitrite, i : 2000 = \ c.c. of 10 per cent, to 100 c.c. of i per
cent. NaCl.
Digitalis, i : 1000 = i c.c. of 10 per cent, to 100 c.c. of i percent. NaCl.
Chloral, i : 1000 = i c.c. of 10 per cent, to looc.c. of i percent. NaC'l.
Barium Chlorid, i : 20,000 = \ c.c. of i percent, to looc.c. of i per cent.
XaCl.
EXERCISE Vm.— (ALL GROUPS) AMYL NITRITE ON CIRCULATION, MAN
(REPORTER III, C)
The circulatory reactions of man may be studied by ordinary clinical
methods, but normal men under the disturbing conditions of the laboratory
i la-- are not good subjects for the usually delicate changes.
Amyl Nitrite, however, gives results which are sufficiently positive. It
i- administered by inhaling ^ drops from a handkerchief,
Experiment i. (Group I) General Symptoms. — Note the bc.uinnii .
duration of the effects. Observe the throbbing of the head; the extent of the
blu>h: the i lian-je- in puUe-rate and respiration.
Experiment 2. (Groups II and III) Blood-pressure. — Observe with one of
the clinical
Tin- pressure in the . ulT is raisol until thr puU- disappears. .m«! then slowly released
until tin- criti.al points are readied. In tin- KonHkow .ius» ult.it. TV method thr stetho-
scope is applied prripln-r.il to thr • tiff, when the following sounds appear successively as
the pressure is released:
like tin- tirst . ardia. -mmd. This indicates the systdif ;
The above sound, with a hissing murmur.
murmur disapjK-ars, ily the sound.
•imd suddenly becomes muffled, and (5) disappears. (4) and (5) indi. ate
the diastolic pressure.
l8o A LABORATORY GUIDE IN PHARMACOLOGY
Experiment 3. (Groups IV and V) Plethysmograph.— Take plethysmo-
graphic tracing.
Experiment 4. (Optional) Sphygmograph.— Take tracing.
QUESTION
Describe the effects of amyl nitrite.
TECHNICAL REFERENCES .
Clinical Blood-pressure Methods. — G. W. Norris, Internal. Clinics, Ser. 25, iv, 61.
Sphygmomanometers. — Tigerstedt, 2.4, 216; Robert, Intox., i, 206; Sahli, 163;
MacWilliam, 1914, Jour. Physiol., 48, Proc. xxviii; Warfield, 1913, Jour. Amer. Med.
Assoc., 61, 1254; Kttotkoff 8 OMJCMftary method: Hirschfelder, Heart; Warfield, 1913, Jour.
Amer. Med. Assoc., 61, 1254; Weysse and Lutz, 1913, Amer. Jour. Physiol., 32, 427.
Physical Factors of Blood-pressure Measurements: Brooks and Luckhardt, 1916, Amer.
Jour. Physiol., 40, 49.
Comparison of methods, Kilgore, 1915, Arch. Int. Med., 16, No. 6.
Posture.— Sanford, 1908, Jour. Amer. Med. Assoc., Feb., 1915.
Excitement. — Zabel, 1910, Muench. med. Woch., 44, 2278.
Sleep and Rest.— Brooks and Carroll, 1912, Trans. Assoc. Amer. Phys., 27, 8.
Pulse. — Robert, Intox., i, 207, 236; Psychic Influence, Lyon and Quails, 1910, Jour.
Amer. Med. Assoc., 55, 455; Polygraph, Stewart, 207; Sphygmo graphs, Tigerstedt, 2.4,
213; Sahli, uo; Venous Pulse, Robert, Intox., i, 240.
Human Blood-flow (and Reflexes). — Stewart, 218; Hewlett, 1913, Arch. Int. Med.,
12, i; n, 507.
Circulation Rate in Man. — Nitrous oxid method of Rrogh and Lindhard, Boothby,
1915, Amer. Jour. Physiol., 37, 383; Means and Newburgh, 1915, Trans. Assoc. Amer.
Phys., 30, 51.
Human Vein-pressure. — Hooker, 1914, Amer. Jour. Physiol., 33; Proc. xxvii; Hooker
and Eyster, 1908, Jour. Hop. Hosp. Bui., 19, 274; A. H. Clark, 1915, Arch. Int. Med.,
16, 587; Influence of Age, Hooker, Amer. Jour. Physiol., 1916, 40, 43.
Human Blood and Plasma Volume. — "Vital red" method; Reith, Rowntree, and
Geraghty, Arch. Int. Med., 16, 547.
Plethysmographs. — The attachment of the cuff to the arm may be sealed with petro-
latum; especially in hairy animals. The plethysmograph may be set on sand, to avoid
vibration. Air transmission gives the best results.
Recording Devices for Plethysmographs. — Besides the usual piston and bellow recorders,
special devices are described by Schlayer, 1906, Cbl. Physiol., 20, 257; Strassburger,
(Spirometer), Arch. ges. Physiol., 139, 33; Dixon, 1907 (frog intestine), Proc. Physiol. Soc.,
Feb. 23.
EXERCISE IX.— (OPTIONAL) ARTIFICIAL CIRCULATION SCHEMA
The effect of changes in the heart and blood-vessels on the blood-pressure and blood-
flow can be demonstrated in an instructive manner by the circulation model depicted in
Fig. 28.
Make the following observations and record them in tabular form. The time can
be kept with a metronome. The pumping should be continued for a short time before
observations are taken.
Students A and B, reading of arterial and venous pressure; students C and D, pump-
ing and outflow; students E and F, recording.
Outflow (at v.).
Arterial pressure. Venous pressure. (Time required
Max. Min. Max. Min. for 100 c c.)
1. (Normal) Pump with moderate excursions;
at rate of 60 per minute. The capillaries-
clamp is partly closed
2. (Vagus Stimulation) Pump at the rate of 10
per minute, allowing complete relaxation,
but incomplete contraction Falls. Falls. Falls.
3. (Vagus Depression) Pump at the rate of 120
per minute, but with very weak compres-
sion . . Little rise. Little rise. Little rise.
CHAP. XXXV
REACTIONS OF BLOOD-VESSELS
181
Arterial pressure. Venous pressure
Max. Min. Max. Min.
4. (Digitalis Action on Cardiac Muscle) Pump at
the rate of 30 per minute, causing complete
contraction, but incomplete relaxation. . . . Rises. Rises.
5. Simultaneous Stimulation of Vagus and Cardiac
Muscle (Digitalis). — Pump at the rate of
30, with complete contraction and relaxa-
tion Rises. Rises.
6. Vasoconstriction. — Repeat i, thin tighten the
iapilhiries-i lamp Rises. Falls.
7. Vasodilation. — Open the capillaries-clamp. . . Falls. Rises.
8. Compldc Digitalic Action. — Combine 5 and 6. Rises more Rises less
than 5 or 6. than 5,
more than
Outflow fat v.).
(Time required
for 100 c c.j
Rises.
Rises.
Falls.
Rises.
Rises less
than 5,
more than
6.
. v- Artihdal (inul-.tion model. The heart i- n-prt-rnted by a rubber syringe bulb
with valves in the direction of th»- ,irr«.\ onpKMed by a lemon-squeezer. The vessels
are formed by rubber Cubing, that for the aorta bein>: I-IK-* Sally da>ti« The arterial pressure is
taken .<>rm-u-r; the vein pressure by an upright tube filled with water. The capil-
lary resistance is furnished by a screw-clamp. The dimensions of the apparatus are indicated on
the figure.
TECHNICAL REFERENCES
Artificial Circulation Schemes, Tigcrstedt, 2.4, 319.
l82 A LABORATORY GUIDE IN PHARMACOLOGY
CHAPTER XXXVI
EXCISED AND FROG HEARTS
INTRODUCTORY
The Heart Muscle. — Aulomaticity. — The cardiac muscle differs from other muscle
by the fact that it contracts rhythmically by an inherent property, /. e., even in the ab-
sence of nervous impulses.1 This property is sometimes called the automatic motor
mechanism of the heart. If the heart is weakened, it may be lost so that the heart may
respond to stimulation by a single contraction, just like ordinary muscle. On the other
hand, the rhythmic property may be imparted to ordinary muscle; for instance, by immers-
ing it in certain solutions of NaCl. The rhythmic property, therefore, does not constitute
a fundamental distinction between cardiac muscle and the other varieties of muscle,
although under normal conditions it is a very important difference. The other properties
of cardiac muscle are still more closely related to those of other muscle: its excitability,
strength of contraction, ton us, etc., may be similarly affected by fatigue or by drugs; in
these respects the myocardium stands intermediate between the skeletal and the smooth
muscle. Normally the rhythmic contractions arise in the base of the heart — in the
auricles, or in the frog in the sinus venosus; and spread gradually to the apex. Conse-
quently the contractions are regular, progressing in a definite order, and all parts of the
heart beat at the same rate, and the two sides of the heart contract at the same time.
The explanation of these facts is that the muscle-fibers at the base of the heart are more
excitable, so that they respond first to the (inherent) rhythmic stimulus; the successive
areas of the ventricles contract then as the result of the stimulus started by the contrac-
tion of the auricles.
Irregularities. — If the excitability of the ventricle is increased as the result of the
action of drugs (such as digitalis, caffein, or aconite), the contraction may start in any
part of the heart. The normal rhythm is thereby destroyed, the contractions cease to progress
regularly, and the rate of each chamber of the heart may differ from the others. If the
contractions arriving from the ventricles coincide with those transmitted from the auricle,
the contractions are strong; if they interfere, the contractions may be weak or absent.
In this way groups of strong contractions may alternate with periods of weak contrac-
tions. A decrease of excitability finds its first expression in the more sluggish ventricles.
As a consequence, a summation of two or more auricular contractions may be necessary
to induce a contraction of the ventricle, and the rate of the latter may be a fraction of
the auricular rate. This is seen with cardiac depressants, especially in the frog's heart.
Another form of irregularity, observed particularly in frogs as the result of digitalis
or aconite, consists in peristaltic contractions, in which the slowly traveling contraction
wave is sharply marked off. This may be due to a quicker contraction, with delayed
relaxation.
Delirium Cordis. — If the cardiac muscle of mammals is overstimulated the contrac-
tions become very irregular. The individual groups of muscle-fibers contract indepen-
dently (hence fibrillary contractions), while the heart as a whole does not perform any
efficient contractions. This condition, also called delirium cordis, appears to be an over-
quickening; it takes the place of the tetanus of the striped muscle, the mammalian heart
being ordinarily unable to enter into tetanus on account of its rhythmic property.2 The
ventricles enter into delirium more readily than the auricles because the latter are cap-
able of a more rapid rhythmic beat, so that overstimulation is not reached so easily.
The frog's heart also does not readily go into delirium because it is too sluggish for over-
stimulation; but when its excitability is raised — as by heat — delirium can be produced.
Since the delirious heart does not keep up an efficient circulation the mammalian heart
(which is nourished by the coronary circulation) is starved and succumbs rapidly to fatigue.
Delirium ordinarily produces paralysis of the heart unless the coronary circulation is sus-
tained artificially. The rabbit's heart may recover spontaneously; the dog's heart does
not.
Coronary Circulation. — The state of the coronary circulation is very important for the
mammalian heart, as its great activity demands a liberal nutrition. The effect is mainly
upon the strength of the contractions, the rate being but little altered. Consequently,
all agencies which depress the heart directly also depress it indirectly by lessening its food
supply, and vice versa. An excessive tonus of the heart by lessening the excursions also
1 In the heart of Limulus (King Crab) and perhaps in some other invertebrates the rhythmic
impulses are generated and conducted by nerves (Carlson, 1904).
1 Tetanus of the mammalian heart can be produced only by the simultaneous stimulation of
the vagus and cardiac muscle.
CHAP. XXXVI EXCISED AND FROG HEARTS 183
starves the heart, so that a strong stimulation of the cardiac muscle may rapidly paralyze
it by interfering with the coronary circulation; and systolic standstill is consequently
impossible in the intact mammalian heart since the starved muscle cannot sustain the
systole.1 A strong contraction of the myocardium also causes a mechanical compres-
sion of the coronary vessels, thereby lessening the blood-flow through them. On the
other hand, extreme dilation of the heart also lessens the coronary circulation; so that an
overdistended heart may often be improved by withdrawing some blood. Since the
coronary vessels possess vasoconstrictor and vasodilator nerves, they may be affected by
drug- acting centrally or peripherally on the vasomotor mechanism.1 The coronary
circulation may also be modified indirectly through changes in the general arterial pressure.
Vasoconstrictors will therefore stimulate the heart,8 and vasodilators will depress it. In
the excised heart these agents may have the opposite effects, since they act then on the
coronary arteries alone; but in the intact animal the effects on the circulation at large will
overcome the effect on the muscle of the coronary arteries.
Changes in blood-pressure have also a mechanical effect on the heart: the cardiac
muscle, like other muscles, contracts better against a certain resistance than against no
resistance. This resistance is furnished by the aortic pressure. The normal blood-pressure
seems to furnish the optimum resistance to the normal heart, so that it would be a mis-
take to consider that a fall of pressure, by lessening its work, would increase the force of a
normal heart. With a wakened heart, however, the optimum resistance falls, so that a
diminished pressure is really beneficial to an exhausted heart.
The amplitude of the contractions is controlled not only by the force of the heart, but
also by its tonus, by its rate, and by the blood-pressure. A tonus which is greatly in-
creased or diminished will prevent the muscle from relaxing or from contracting to the
usual extent. An increased rate does not allow time for complete contraction and relaxa-
tion, and so renders the beats more shallow, while a slow rate tends to increase the ex-
cursions. A high blood-pressure prevents the complete emptying of the heart, and thereby
renders the beats more shallow and slows the rate. (In intact animals this slowing is
very marked, being due to a reflex stimulation of the vagus mechanism.)
The volume of blood thrown out at each beat varies with the amplitude of the excursions.
The output in a given lime is the product of the rate and the volume of each beat. The
•work done by the heart is the product of the output and the resistance (blood-pressure)
against which it acts.
.' of the Rate of the Heart on the Output. — The output of the heart is greatly di-
minished by slowing its ordinary rate; the increased volume of each beat being insufficient
to counterbalance the lessened number of contractions. A quickening of the heart above
the normal, on the other hand, causes but little increase in the output, since the lessening
of the volume of each beat nearly offsets the increased rate. This, as well as the effect
of the vascular system, etc., may be demonstrated on an artificial circulation apparatus
The Innervation of the Heart.— Although the cardiac muscle is able to perform regular
rhythmic contractions in the absence of nerves, it is normally under nervous control.
Besides the sensory (depressor) nerve there are two motor nerves, the vagus and tl
celerator branch of the sympathetic. The origin (center) of both of these nerves i> in the
medulla. Both nerves run in the same sheath in the frog, but are separated in mammals.
Both nerves are connected with ganglia. Those of the \agu> are contained in the heart
itself (in the frog these vagus ganglia are situated especially at the juncture of the >inu>
venosus with the auricle). Those of the accelerator are extracardiat . and in mammals
lie probably in the inferior cervical and in the stellate ganglia, around the sub.laxian
artery. The endings in the cardiac muscle are "free ending-." similar to those of un-
striprd mils. lr. The heart contains no structures corresponding to the end-plates of
1 muscle.
The effect of electric stimulation of these nerves appears onl iight latent
period, and disappears after a time, even it" the stimulation i> continued. The latent period
and the action are longer for the vagus.
Cfl a -lowing of the rate, the diastole being i>pe( ially
prolonged. The irritability and the lontra. tile |xmet are in< rr.i-cd in mammals the
amplitude of the ext ur-i"i . the tonus is diminished; the blood p : out-
put fall. Strong stimulation causes diastolic standstill
1 The man; however, capable of (tystolic standstill if the coronary cinul.ition
is mar i method
»In workii. . perfused hearts it i- imj>ort.int to remrml.er th;it the »
vessels are also affected by the trmprraturc nt thf Mood. !•< n..- .hlv v.l l>\ cold.
i the cardia. . It. lu-.it iiim-kming tli,
while cold Slow* thr • ontr.u tioOS.
•Rhythmical \w.\- ! mammalian heart l.y -imply raising the
nt gases (hydrogen, Magnus, 1002) or <
184 A LABORATORY GUIDE IN PHARMACOLOGY
Accelerator stimulation (the anterior ramus of the annulus of Vieussens) quickens
the rate, shortening all the phases except the auricular systole and the ventricular dias-
tole. The excitability, tonus, and strength of the heart are increased, but the pulse is
more shallow in mammals (in the frog the excursions are also increased). The blood-
pressure and the output rise somewhat, but not commensurate with the increased rate.
Tonic Impulses. — The vagi are tonically active in some animals (notably in man and
in dogs), so that division of these nerves causes a quickening of the heart. In other
animals (rabbits) there is normally no tonic action, so that division produces little effect
on the heart rate. The accelerators are also tonically active, but division of these nerves
produces less effect than section of the vagi. The ganglia and endings also have some
tonic action, for a further quickening may be obtained, after section of the vagi, by paral-
yzing the vagal endings.
The vagi or accelerators may be stimulated or depressed directly at their origin, or
in any part of their course, by drugs or by other means. They can also be affected indi-
rectly, especially the vagus. A fall in blood-pressure, most forms of reflex irritation, mus-
cular exercise, swallowing, etc., quicken the heart by inhibiting the vagus center. Stimu-
lation of the trigeminal endings, on the other hand, excites the vagus center and slows, or
even stops, the heart. A rise of blood-pressure also stimulates the vagus and causes
slowing.
Methods of Studying the Actions of the Heart. — It follows from the preceding that
the action of the heart depends on a considerable number of interrelated factors. These,
acting together, produce the phenomena which may be studied on normal animals by the
pulse and apex-beat; and on operated animals by direct observation and tracings of the
exposed heart. The intracardiac and the general blood-pressure and the output of the
heart, etc., are also determined in large part by the cardiac activity; but since they also
depend on the state of the vasomotor system, they must be supplemented by more direct
methods. Indeed, all the observations on intact animals give only the sum of the factors
which may be involved. It is evident that no understanding of the action of a drug is
possible until the share of each factor is known. This must be determined by isolating it
as completely as possible from the other factors.
Suitability of Different Animals. — The hearts of frogs and turtles are convenient for
studying the effects of drugs, since they continue beating normally for a considerable time
after they are exposed or excised. Many phenomena can be observed very well by
direct inspection or by perfusion, others may be recorded by levers, etc.
The cardiac nerves of frogs are also situated conveniently. It must not be forgotten,
however, that the physiology of the heart of cold-blooded animals differs considerably from
that of the warm-blooded; and caution must be used in applying the results obtained with
the one to the other. The main uses of the frog's heart are, therefore, restricted to pre-
liminary studies, to the investigation of special problems, and to the convenient demon-
stration of actions which have been already controlled on warm-blooded animals. Among
the latter the functions of the myocardium are identical, as far as we know. The absence
of tonic vagus impulses in rabbits must be borne in mind.
Drugs may act on the heart in three ways: (i) Directly on the cardiac muscle; (2)
directly on the cardiac nerves, and (3) indirectly, on either the muscles or nerves — through
reflexes, altered resistance, altered nutrition, altered coronary circulation, etc.
Methods of Studying the Direct Effects on the Cardiac Muscle. — These demand that
the resistance to the work of the heart be kept constant — an object which can only be
accomplished by separating the heart from the general vascular system. The pulmonary
circulation may generally be kept intact, as it is not much affected by drugs. The methods
of isolating the heart may, however, be conveniently divided into those which retain the
pulmonary circulation and those which do not." The nervous mechanism should also be
excluded. If it is desired to retain the intracardiac nervous apparatus, it suffices to cut
the trunks of the vagi and accelerators, or to shut off the blood from the medulla. The
intracardiac vagus mechanism can also be paralyzed by atropin. This leaves only the
accelerator endings.
The frog's heart will continue to beat for some time after it has been excised from the
body; but the mammalian heart requires that the coronary circulation be maintained.
This may be done by the heart itself, or by injecting the perfusion fluid under pressure.
Perfusion Liquids. — In perfusing the excised heart a fluid must be employed which
does not produce any salt or ion action, which contains oxygen and nutriment, and which
is at body temperature. The best is oxygenated defibrinated blood from the same species
of animal, diluted with 5 volumes of Locke's solution. Other fluids may be substi-
tuted, but these must be charged with oxygen when used with the mammalian heart.
Serum may be employed. An excellent substitute is Locke's Fluid. By the use of
Langendorff's or Porter's method the heart can be kept beating, or revived, many hours
after death.
CHAP. XXXVI EXCISED AND FROG HEARTS 185
Similar solutions may be used for the perfusion of frogs' hearts, except that they
should contain less salt (0.6 to 0.75 per cent. NaCl). Used alone, this saline solution
gradually poisons the heart after the manner of digitalis. The toxicity is less if 2 per
cent, of gum arabic is added, or small quantities of some other salts. Ringer's Solution
(see Index) has been found very good. Rabbit's or beef's blood, defibrinated and diluted
with 2\ parts of 0.6 per cent. NaCl, is also used.
Analysis of the Effects on the Heart. — Actions on the nervous mechanism can be
studied with the heart in situ by dividing or stimulating the vagi and accelerators at
different levels. I' yon has also devised a method of studying the effects of drugs upon
the cerebral cardiac centers by separating these from the general circulation and arti-
ficially i irculating through them defibrinated blood containing the jxrisons to be studied;
in this way they do not reach the heart at all.
An effect upon the nerves is manifested particularly by changes in the rate of the
heart; but as the rate may also be modified through the muscle, or indirectly, a more
detailed analysis becomes necessary; this will repay closer study, as it illustrates the
methods of pharmacologic research.
Investigation of Changes in the Rate of the Heart. — Quickening may be due to a
: or rellex inhibition of the vagus, or to stimulation of the accelerator nerves or of the
cardiac muscle.
(i) If the quickening does not occur after section of the vagi, it must have been due
:!ral paralysis of lite vagi. If the center does not respond to reflex stimulation (such
as the inhalation of ammonia with rabbits), the center it<df is paraly/.ed. If it does
respond, the inhibition of the vagus must be reflex, which can be further demonstrated by
division of the corresponding path.
If the quickening occurs after section of the vagi, the drug is tried on animals
in which the vagus endings have been completely paralyzed by atropin. If it produces
:iect, the drug must paralyze either the ganglia or endings. It is tried on animals
in which the ganglia have been paralyzed by nicotin; or on the ganglion-free apex of the
frog's heart. If it produces no quickening, it must have paralyzed the vagus ganglia; if
quickening occurs, it must paralyze the endings. In the former case, stimulation of the
Minis, in the frog, stops the heart; if the endings are paralyzed stimulation of the sinus
10 effect.
(3) If the quickening occurs even after atropin, there must be a stimulation of either
the accelerator mechanism or of the cardiac muscle. If the effect occurs on the excised
atropini/.ed heart, it must stimulate either the muscle or the accelerator endings. It is very
difficult to distinguish between these; the study of the relative duration and strength of
the phases of the cardiac cycle furnishes some indication. The cardiac muscle, quite free
from nerve-endings, can also be studied in the embryonal chick. It appears, from these
methods, that the stimulation is always of the muscle rather than of the endings, so that
we -hall designate a quickening obtained after atropin as a stimulation of the card
It the drug acts after atropin, but has no effect on the exc i-ed heart, it must
stimulate the accelerator center. This ian l>e further shown by its producing no effect on
the intact animal if the spinal cord is divided above the first dorsal vertebra, or if both
Mellate gamrlia are excised.
Slowing may be due to direct or reflex stimulation of the v. . ly-is of the
rators to paralysis of the muscle, direct or through impaired nutrition; or to systolic
stimulation of the muscle.
i. If the slowing does not occur after se< lion of the vagi, it must be due to a stimulation
of Hit vagus center, espe< ially if de« trie simulation of the vagus trunk continues etTc
i of the vagi, but not after nicotin. it must be due to stimula-
tion of the vagus ganglia.
of the vagi and after nicotin. but not after atropin. it nui>t
be due to stimulation of the vagus ending. Kle.tri. simulation of the vagus trunk i<
tropin. but not after division of tin I l>e due
imilation of the accelerator nerve is effec-
tive, the depression must be central; if not. it i^ peripheral.
5. If Mtcr atropin and after division of the accelerators, it must be due to
a direct action on the i ardi.n imi- le. or to insufficient nutrition. The latter may be ex-
• hided by ir« ul.ition. If the slowing per-i-t-. it i- due to far
creased tonu^ of the <ardia. muscle; the strength of the contra- tions will induatc \\hi< h
i- the true explanation.
Cardiac standstill may be due to stimulation of the vagus, to paralysis of the cardiac
muscle trogs) to excessive sy
the standstill disappears on section of the vagi, it is due to stimulation of the
vagus center.
i86
A LABORATORY GUIDE IN PHARMACOLOGY
2. If it persists, but disappears after atropin, it is due to peripheral stimulation of the
vagus. The ganglia and endings can be distinguished as in "Slowing," 2, 3. The frog's
heart is strongly diastolic if the stoppage is due to stimulation of any part of the vagi.
3. If atropin does not relieve the standstill, it is caused by a direct ejfect on the muscle.
In mammals, this is always paralytic. In frogs it may be due to paralysis, when the heart
is of medium size, and cannot contract if it is forcibly distended; or to excessive systole
(Digitalis group) when the heart is very small, and contracts if distended.
4. The paralysis may only involve the rhythmic power, so that the heart responds to
stimulation (i. e., a pin-prick) by a single contraction; or it may be complete.
EXERCISE I.— (DEMONSTRATION) EXCISED MAMMALIAN HEART
(LANGENDORFF METHOD)
(REPORTER II, A)
The method consists essentially in perfusing the coronary vessels of the
excised heart with a warm oxygenated saline solution. Various arrange-
ments may be used, the following being one of the simplest, but not sufficient
for exact work :
Perfusion Apparatus (See Fig. 29). — A large water-bath, iv.b., heated by a Bunsen
or alcohol burner, is arranged on a shelf 150 cm. above the table. In this is set a 2-gallon
/ M
Fig. 29. — Apparatus for Langendorfi heart (see text).
bottle containing 7 liters of fresh Locke's fluid. Into this bottle dips a siphon, a narrow
orifice tube connected with the oxygen-tank, and a thermometer. The siphon tube is
prolonged to the table. A T piece, t'1, is inserted near the lower end, the free limb being
* This serves for the removal of air or of cooled blood, if the flow has been arrested.
CHAP. XXXVI EXCISED AND FROG HEARTS 187
closed by a Mohr clamp. The tube terminates in another T, /", which bears the bulb
of a thermometer. This T is joined to the aortic cannula, and supported by a clamp and
stand, over the hot-water funnel /. This is kept warm by a Bunsen or alcohol flame.
A pin is hooked to the apex of the heart, h, and connected with a string, which passes
through the stem of the funnel to a muscle lever, m.l., writing on the drum d. The lever
i^hted with a lo-gm. counterpoise. It is best to attach the string to the lever with a
pin. so that the excursions can be regulated to i J or 2 inches. A beaker is set beneath the
funnel to catch the blood. Several drums should be smoked in advance. The whole
apparatus should be ready before the heart is excised.
Preliminary Operations. — While the apparatus is being set up the dog is anes-
thetized, and cannulae are tied in the trachea, carotid, and femoral vein. The latter is
connected with the injection buret. The dog is now bled from the carotid as long as the
blood-flow is a strong stream. The carotid is clamped, and the blood is defibrinated,
>trained, and heated to 45° C. and poured into b. The heart is now exposed and arti-
tu ial respiration is started. The carotid is again opened, and the dog is bled, while at
the same time a liter of warm Locke's fluid is allowed to flow into the femoral vein.1 The
diluted blood is collected as long as it flows from the carotid, defibrinated, strained, mixed
with the blood which was previously drawn, heated to 45 ° C., and poured into the reservoir.
The reservoir is now shaken so as to mix the fluids, and a slow stream of oxygen is
passed through it. The siphon tube is filled with the blood.
In the meantime the heart is excised with an inch of the aorta, and with the lungs.
The latter are trimmed away, the pericardium is slit open. All branches of the aorta are
tio«l. The aortic cannula is introduced and secured by a firm ligature, taking care that
it does not interfere with the play of the semilunar valves. The aorta is clamped below the
cannula; this is filled with blood, connected air-free with /', and supported in the clamp.
The pin is hooked into the apex, connected with the lever, the clamp on the aorta is
removed, and the perfusion is started. The pressure closes the semilunar valves, so that
the fluid is forced through the coronary circulation, escaping through the right auricle
and into the beaker. The flow should be rather free, the beaker being frequently ex-
changed, the unpoisoned blood being returned to the reservoir. If it is too free, some of
the veins may be closed by bulldog forceps. See that the thermometer in /" registers
38° to 42 °C.
The heart will begin to beat in a very short time, at first feebly and irregularly, but
soon with strong, regular beats. The observations and tracings may be started at this
time. The solutions should be injected just below /' with a hypodermic syringe, thrust
obliquely through the rubber. The injections should be made very slowly, and continued
until the desired effect is obtained.
cad of injecting the drugs with a syringe, they may be added directly to the
perfusing fluid, in the proportion of about i : 25. A second reservoir will be necessary.)
Experiments. — i. Strychnin. — Obtain a normal tracing. Inject i : 5000
Strychnin. According to the dose (which is really inversely proportional
to the rate of perfusion), one may obtain:
\M effect.
(6) Increased excursions.
(c) Diminished excursions.
2. Caffein. — Inject i : 5000 Caffein. According to the dose, one may
obtain quickening and increased excursions; or slowing with diminished
excursions.
3. Chloroform. — Inject a saturated solution of Chloroform in normal
saline: the heart is slowed and especially weakened. Proceed to 4 before it
has quite stopped.
4. Epinephrin.— Injn t i : 10,000: the heart revives promptly and beats
powerfully.
5. Potassium.— Inject i : 100 KC1: sudden paraly>i< of the heart.
Recovery may be s|xmtaneous or occur by 6, which should be undertaken
6. Camphor. Inject a saturated solution of Camphor in normal >aline
solution: t Mhened.
••MIT. Jour. Physiol., 36, ijo, claim that the heart behaves much
better if it is excised without previous bleeding.
l88 A LABORATORY GUIDE IN PHARMACOLOGY
7. Digitalis. — Inject i : 100: the heart is first quickened and strength-
ened. The tonus increases. Finally it goes into delirium cordis and stops
in systolic position.
(Optional) Concentrations of Various Drugs for Direct Perfusion (Greene). — Aconite,
0.0002 per cent.; Alcohol, 0.4, i, and 2 per cent.; Atropin, o.ooi per cent.; Caffein, 0.02 to
0.5 per cent.; Chloroform, 0.02 to 0.05 per cent.; Digitalis, o.oooi and 0.0005 Per cent.;
Ether, i per cent.; Morphin, 0.5 and i per cent.; Nicotin, o.ooi per cent.; Physostigmin,
o.oi per cent.; Strychnin, 0.005 ancl °-02 Per cent.; Yeratrin, 0.002 per cent.
TECHNICAL REFERENCES
Fuller descriptions are given in Langendorff, 1895, Arch. ges. Physiol., 61, 291;
Stewart, 203; Pittenger, 126; Tigerstedt, 2.4, 144; Robert, Intox., i, 180; Greene, 73;
Abderhalden, 3, 333 (metabolism, ibid., 374).
The following modifications and improvements may be mentioned: Herlitzka, 1905
(Constant Pressure), Arch. ges. Physiol., 107, 564; Locke and Rosenheim, 1907 (Con-
tinued Perfusion with Small Quantities), Jour. Physiol., 36, 205; Brodie and Cullis, 1908
(Uniform Temperature and Small Dead Space), Jour. Physiol., 37, 337; Eyster and Loewen-
hart, 1913, Jour. Pharmacol., 5, 57; Dresbach, 1913, Quart. Jour. Exp. Physiol., 8, 73;
A. I. Gunn, 1913, Jour. Physiol., 46, Aug. 18 (good arrangement for heating the injection
fluid).
Technical Notes. — Outline of Methods for Studying the Isolated Mammalian Heart. —
The methods which have been employed for the study of the isolated mammalian heart
are briefly as follows:
I. Methods Employing the Whole Heart and Pulmonary Circulation (Excluding the
Peripheral Yessels and, to a Large Extent, the Brain — Fig. 30). — These methods differ
by the manner in which the action of the heart is observed or recorded, which may be
done by direct observation, by taking pressure curves from the carotid or from the ven-
tricles, or by the myocardiogram. The methods consist essentially in establishing a
connection between the large arteries and large veins, and then ligating the vessels periph-
erally to this connection. The vessels which are employed for this purpose and the
apparatus used for establishing the connections vary in the different methods.
(A more recent "isolated lung-heart preparation" for dogs is described by Knowlton
and Starling, 1912, Jour. Physiol., 44, 206; 45, 146.)
(a) Communication established between the aorta and right auricle:
1. Martin's Original Method.. — In this a' communication is established through a
reservoir containing defibrinated blood and connected with the right auricle, while the
left ventricle pumps the blood through a tube back into the reservoir. The course of this
blood then is: right auricle, pulmonary circulation, left heart, standing tube, and reser-
voir. The oxygenation of the blood is effected by artificial respiration.
2. The modified method of Martin and Applegarth establishes a communication through
the coronary vessels, the maintenance of pressure being aided by connection of the aorta
with a reservoir containing defibrinated blood. The course of the blood is: aorta, coro-
nary circulation, right heart, lungs, left heart, and aorta. Oxygenation is by artificial
respiration.
3. The McGrath and Kennedy method is an amplification of the last, in that it measures
the intracardiac pressure and the outflow through the pulmonary artery.
4. Hedon and Arrous' method differs from the preceding methods by leaving out the
reservoir, simply tying the aorta and its branches and the vena cava. The course of the
blood is: aorta, coronary circulation, right heart, pulmonary circulation, left heart, and
aorta. Oxygenation is by artificial respiration.
The heart survives some hours. It becomes progressively slower by the using up of
material and the production of waste products, but it remains regular.
5. Cyon connects the aorta with the vena cava. In addition, he is very careful to
ligate all the vessels leading to the brain, so that he can expose this organ to poisons
without their reaching the general circulation.
(6) Communication Through the Carotid and Jugular. — The methods differ mainly
in the mechanism introduced as resistance, this being either constant or variable:
1. Stolnikow makes the connection through two glass vessels of known content, which
are reversible, and one of which is alternately filled by blood expelled from the heart,
while the other empties into the vena cava. In this way the volume of blood expelled
by the heart in a given time can be measured. The other vessels are, of course, ligated.
Oxygenation is by artificial respiration.
2. Bohr and Henriquez establish the connection by a simple tube. Hering does not
ligate the veins, using them as a pressure regulator. Bock forms the connection through
CHAP. XXXVI
-MD AND FROG HEARTS
I89
a compressible tube and screw-cock, so that a varying resistance may be introduced.
He describes a rather complicated improvement of the method in Arch. exp. Path. Pharm.,
1908, Suppl., 83.
In all these methods the registration is done by a manometer in the other carotid,
the aorta and vena cava being ligated and artificial respiration being kept up.
II. Completely isolated hearts, /. «•., without the pulmonary circulation, but with
the ganglia -till active. In these methods the blood must be artificially oxygenated, and
is usually introduced under pressure. Otherwise the methods are similar to the preceding.
Trtssurt
Martin and Applegarth. Tschl-
tou-itch connects the pulmonary
artery and vein by a tube.
Martin's original method. Bock.
Fig. 30.— Methods of studying the isolated mammalian heart.
;.r.n li..illy Martin'- original mrtli<»! - the pulmonary
artery with the pulmonary vein ; the coune of the blood being: reservoir, jugular
vrin. ri^ht In-art, nmnoting piivi-. It-It heart, .mrta. atnl reservoir.
ma <>nl\ the ...p. nary « in illation. intnMhu in« the blood int<> tl;«
whi. h it KIH-S thnmjih t!
of tin- ri^'lit In-art. Tin- ^liaju- «>f tin- In-art, luiinlu-r ami -trench ••! Krat-. atnl tin- number
of drop- tlouini: through tin- rii:ht ! - i-un-o! in thi- way.
I90
A LABORATORY GUIDE IN PHARMACOLOGY
3. Hedon and Arrous ligate the aorta and vena cava and connect the pulmonary
artery and pulmonary vein directly, feeding the heart with its own blood and keeping it
alive by artificial methods.
4. Ileymans and Kochmann connect the aorta of the excised heart with the carotid
of a second animal, letting the blood return through a funnel connected with the jugular;
or without the use of a funnel, by connecting the pulmonary artery of the excised heart
with the jugular of the animal, and tying the other vessels.
III. Isolated apex preparations, /. c., ganglion-free heart muscle. Porter has suc-
ceeded in maintaining rhythmic contractions of isolated strips of the apex of the heart by
injecting oxygenated blood under pressure into a branch of the coronary artery supplying
it.
The methods of Langendorff and Porter have been criticized as yielding abnormal
results, because they leave the cavities of the heart empty. Their results must, therefore,
be interpreted cautiously. Gottlieb and Magnus (1903) obviate this difficulty by filling the
ventricle with a distensible balloon.
Technical References. — Heinz, i, 184; Meyer and Gottlieb, 202.
EXERCISE H.— (DEMONSTRATION) PERFUSION OF FROG'S HEART
(REPORTER IV, C)
The frog's heart may be perfused either through the sinus venosus and
auricles or through the aorta and ventricle.
Technical References. — The various frog methods are discussed in Abderhalden, 3,
329; Robert, Intox., i, 177, 193; Tigerstedt, 2.4, 123; Greene, 69.
Experiment i. (Optional) Demonstration of Williams' Apparatus. — This has been
used extensively in pharmacologic work on the frog's heart, as it permits the study of a
number of phenomena under a variety of conditions. An artificial circulation is main-
tained through the ventricle by means of solutions, to which the poisons may be added.
The apparatus (Fig. 31) consists of a reservoir and a system of tubes provided with slit
valves ( V and V) and a two-way cannula.
These allow the perfusing liquid to get into
the heart ( H) and to be pumped in a definite
direction. The cannula is introduced through
the bulbus aortae into the ventricle and tied.
(The apex of the ventricle may be used
alone:) Each contraction of the ventricle
forces the blood through V into the up-
right tube, and from here into the reservoir.
The relaxation of the heart allows the liquid
to enter from V. The auriculoventricular
valves prevent the blood from coming back
into the auricle. The number of drops
flowing into the reservoir can be counted,
and give an idea of the work done. By
raising or lowering the reservoir the intra-
cardiac pressure can be varied;2 by applying
the screw-clamp beyond V one may intro-
duce resistance; by clamping this tube alto-
gether and opening communication to a
small mercury manometer the absolute press-
ure can be measured and tracings taken.
The changes in volume, corresponding to the
extent of the excursions, may be read from
the millimeter scale, MS.
Technical References. — Williams, 1877, Arch. exp. Path. Pharm., 13, n; Dreser, 1888;
ibid., 24, 223; Rothberger, 1907, Arch. ges. Physiol., 118, 353 (Work of Heart).
Experiment 2. (Demonstration) Perfusion of Ventricle by Straub-
Fuehner Method. — This consists in introducing a suitable cannula through
the aorta into the ventricle of the excised heart.
1 The valve V should point in the reverse direction. Fresh frog's skin is convenient for these
valves.
* A pressure of 200 mm. of water is the optimum.
Fig. 31. — Williams' heart apparatus.
CHAP. XXXVI
EXCISED AND FROG HEARTS
IQI
The preparation may be used to demonstrate the effects of the following
drugs:
(a) Calcium (Straub, 1912).— A tracing is taken with ordiqary Ringer's
Fluid. This is then replaced by Ca-free Ringer's: the excursions become
very weak. Replace by ordinary Ringer's: the heart recovers. Replace
by Ringer's containing CaCh 0.8 : 1000 (instead of the normal 0.2 : 1000):
the excursions are again diminished. A definite ratio of Ca is therefore
necessary for functionation. (The calcium does not penetrate into the
muscle, but acts on the cell membrane.) Replace by ordinary Ringer's
solution and let conditions return to normal.
Question. — Describe the action of calcium on the heart muscle.
(b) Potassium. — Use 0.5 c.c. of 10 per cent, in 10 c.c. of Ringer's.
(c) Strychnin. — Use i c.c. of i : 1000; then i c.c. of i : 100 in 10 c.c. of
Ringer's.
(d) Cajfein. — Use i c.c. of i : 100 in 10 c.c. of Ringer's.
(e) Aconitin. — Use i c.c. of i : 10,000 in 10 c.c. of Ringer's.
(/) Epincphrin. — Use i c.c. of i : 10,000 in 10 c.c. of Ringer's.
Questions. — Describe the effects of these drugs. Which are stimulant
and which depressant to the heart?
'.-/«;/ Notes. — The method was described by Straub, 1910, Bioch. Zt., 28, 394,
and modified by Fuehner, Narhweiss, 123, as follows:
Large frogs (60 to 100 gm.) should be used. A small dish
with Ringer's solution and containing the cannula (Fig. 32)
should be at hand; also ligatures and dissecting tools.
Decapitate the frog, leaving the lower jaw, and pith the
spinal cord. Lay frog on plate, head toward operator. Lift
the skin of the throat with forceps and cut away a wide flap
of skin over the thorax, reflecting it down over the abdomen.
Rinse the scissors and split the sternum, from above down-
ward, to the abdominal muscles, where the opening is enlarged by
a transverse incision. Cut away the sternum on both sides to
the arms. Turn the plate about so as to bring the feet toward the
operator.
Fig. 32.— Fuchner1* heart cannula,
actual size (Fuehner).
v— Insertion of aortic cannula (Fuehner).
Slit tin- |.«-rii .inliut; !'!.«« «' •»
ligature around the aorta beyond it- hit ur. at i"n and loop i it <!<> not i
IQ2
A LABORATORY GUIDE IN PHARMACOLOGY
one of the branches and insert the cannula, containing a little Ringer solution, and very
carefully push it into the ventricle in the direction of the arrow in Fig. 33. This is a
rather delicate operation. Force must be avoided, the cannula being slipped gently to
and fro, toward the back and left side of the frog, until it enters during a systole (practice
on dead frog).
The cannula enters rather suddenly, necessitating care that it does not slip back.
Tighten the ligature, making sure that the aorta has actually entered by carefully feeling
the ventricle and observing the movement of the blood in the cannula. Remove this
with a pipet, the point of which should come to, but not enter, the heart (Fig. 34). Rinse
with Ringer's until the solution remains blood free. Raise the cannula and excise the
heart, dividing successively the aorta, frenulum, and cavae as far as possible from the
sinus. It is rather advantageous to tie the vein before dividing
(Fig. 35)-
The apex of the ventricle is now gently clasped with a deli-
cate rather broad-pointed clamp (Fig. 36). (Mendenhall, 1915,
Jour. Pharmacol., 6, transmits the movements through a small
tambour connected with the top of the cannula.) Fasten the
heart-cannula in a moist chamber, through which oxygen is bub-
bling, and connect tor tracings (Fig. 37).
Fig. 34. — Pipet for heart cannula,
reduced size (Fuehner).
Fig. 35-— Excision of heart (Fuehner): a, Ventricle;
b, auricles; c, sinus venosus; d, vena cava.
Experiment 3. (Optional) Heart Perfusion in Bio-assay. — The Straub preparation
may be used for Digitaloids, Aconitin, and Muscarin (Fuehner, Nachweiss, 128; Proof of
Aconitin, ibid., 102 and 130; Arch. exp. Path., 1911, 66, 179.
Experiment 4. (Optional) Perfusion of Auricles and Ventricle by Hartung's Method. —
Arch. exp. Path. Pharm., 1911, 66, 3.
This maintains a circulation through the entire heart. A. J. Clark, 1912, has intro-
duced some slight modifications, Proc. Roy. Soc. Med., 5, 181. Simpson, 1911, Quart.
Jour. Exp. Physiol, 4, 249, describes a cardioplethysmograph. Santesson, 1915, Nord.
Med. Ark., prefers perfusion through the vena cava in situ.
The apparatus may be used to demonstrate the action of aconitin i : 100,000.
Experiment 5. (Optional) Perfusion of Frog's Heart in Situ. — The most convenient
method consists in placing cannulae into the ascending vena cava and in one of the aortae,
both pointing toward the heart, and ligating the other vessels. The vein cannula is
connected by rubber tubing with a Mariotte bottle. Air-bubbles must be rigorously
excluded. The aortic cannula is also connected with a tube, through which the fluid
can return to the reservoir. The latter is filled with the perfusing fluid — Ringer's solution.
By raising the reservoir the diastolic pressure can be varied at will — 4 to 6 cm. gives the
best results. The resistance to the heart can be varied by raising or partially clamping
the aortic tube.
The observations are made by counting the number of beats and the outflow per
CHAP. XXXVI
EXCISED AND FROG HEARTS
193
minute. Tracings may be taken by any of the methods. The heart may be left in the
body or excised.
If drugs are to be perfused, it is well to connect two reservoirs with the vein cannula.
Any of the following drugs may be used (Greene): Alcohol, 2 to 5 per cent.; Caffein, o.i
and 0.2 per cent.; Calcium Chlorid, 0.03 percent.; Chloral, 0.2 per cent.; Chloroform,
Fig. 36— Isolated heart
(Fuehner).
Fig. 37.— Heart-chamber (Fuehner).
0.05 per cent.; Ether, i per cent.; Morphin, 0.5 per cent.; Quinin, 0.05 and i per cent.;
rin, 0.005 per cent.
Experiment 6. (Optional) Isolated Auricle of Frog. — Sn \\ . StrauK Arch. exp.
Path. I'harm., 70, 19.
EXERCISE in.— BIO-ASSAY OF HEART TONICS: DIGITALIS, ETC.
-•RTKR IV. C)
Introductory Discussion of Bio-assay. — The natural variability of bo-
tanic and animal drugs and their deterioration on keeping, etc., necessitate
the determination of thc-ir strength, especially in tin potent <1
Chcmic assay of the active constituents when possible is preferred. 1!
ever, when a drug contain- *everal active constituents, and particularly
when the>e are not identified, chemic assay is not feasible. In such cases
tin- activity may be estimated through comparative experiments on animals,
by determining the dose required to produce some definitely ;isc ertainable
pharmai < When a drug contains several ingredient* produc-
ing rather different effect* the test should refer to the action which is CS-
dly utili/ed in therapeutic*. Uccau*c of the- variability of bio!
reaction- the- re*ult* arc- not usually a*- accurate as are the better ch-
method*, but they UK at lea*t much better than nothing. The- method*
should. BO far as possible, exclude marked personal factors in tec hnic or in
inter] »
The II based on the card; no! the drug.
They dlffei mainly in the convenience of their application.
'3
IQ4 A LABORATORY GUIDE IN PHARMACOLOGY
Technical References. — Pittcnger; Fuehner; H. C. Wood, Jr., 1912, Jour. Amer.
Med. Assoc., 59, 1433; Philadelphia Commission, 1911, Amer. Pharm. Assoc., Bui. 6, 22;
Houghton, IQII, Amer. Pharm. Assoc., Bui. 6, 176.
General Discussion of Digitalis Methods. — Hale, 1911, Hyg. Bui., No. 74; Pittenger;
Heinz, 1913, Merck's Rep., 26; Holste, 1914, Zs. exp. Path., 25, 385; Barger and Shaw,
1904, Yearb. Pharmacy; Santesson, 1915, Nord. Med. Ark.
Experiment i. (Demonstration) Official Frog Method (U. S. P. IX).—
Exact graded doses are injected into the ventral lymph-sac of weighed frogs
(best between 20 and 30 gm.). These are pithed at the end of one hour
(Famulener and Lyons method) or of twenty-four hours (Houghton
method). The two methods give very similar results. The heart is ex-
posed and inspected. The end-reaction is definite arrest of the ventricles
in systole with the auricles dilated. The dosage which just suffices to
produce this effect corresponds to "M. F. D." (minimum fatal dose). With
digitalis this should correspond to about 0.6 mg. per gram of frog.
Since this dose varies not only with the sample of the drug, but also
with the species of the frogs, the season, and other uncontrollable condi-
tions, the sample to be tested must always be compared with a sample of
known activity.
Ouabain (crystallized strophanthin)1 is used for this purpose. The
ordinary M. F. D. of this is about 0.00045 m£- Per gram of frog.
For exact work the details of the official process must be consulted.
The method may be demonstrated by injecting three weighed frogs (about
20 gm. weight) with a i : 50,000 solution of ouabain (i c.c. = 0.02 mg.),
giving respectively 0.25, 0.5, and 0.75 per frog (corresponding to 0.00025,
0.0005, and 0.00075 mg- Per gram, respectively). After an hour the frogs are
opened. The ventricle should be beating in the animal with 0.25 c.c.,
arrested in that with 0.75 c.c.; that with 0.5 c.c. may be either beating or
stopped.
Dilute some Tr. Digitalis with an equal volume of water, and inject into
three frogs, using respectively 0.2, 0.4, and 0.6 c.c. of the dilution per 20
gm. frog, corresponding to 0.5, i, and 1.5 mg. of Digitalis per gram. From
the M. F. D. calculate how much Digitalis corresponds to 0.0005 m£- °f
Ouabain.
The following doses are accepted as equivalent to the standard dose of ouabain
(0.0005 mg- Per gram of frog):
Dose (gm. or c.c.
Preparation. per gram of frog).
Digitalis 0.0006
Tincture 0.006
Strophanthus 0.000006
Tincture 0.00006
Convallaria 0.00018
Apocynum • 0.00005
Squills 0.0006
Question. — Describe the principle of the official Digitalis Assay.
Technical References. — U. S. P. IX; Hale and Service, 1911, Amer. Jour. Pharm., 83,,
97; Hale, IQII, Hyg. Bui., No. 74; Hamilton, 1912 (Heart-tonic Unit, H. T. U.), Amer.
Jour. Pharm., 84, 97; Houghton and Hamilton, 1909, Amer. Jour. Pharm., Oct.; Houghton,
1909, Lancet, June 19; Rowe, 1915 (Comparison One- and Twelve-hour Method), Jour.
1 Houghton proposed crystallized kombe strophanthin as standard (Amer. Pharm. Assoc.,
Bui. 6, 176), but this has not been accepted.
CHAP. XXXVI EXCISED AND FROG HEARTS 195
Amer. Pharm. Assoc., 4, 108; Committee, Jour. Amer. Pharm. Assoc., 1912, i, 1305;
Gottlieb, 1914, Muench. med. Woch., 813.
Minimal Fatal Dose. — Houghton, 1909, Lancet, June 19.
E/ect of Temperature. — Sollmann, Mendenhall, and Stingel, 1915, Jour. Pharmacol., 6.
'.•/ Fluctuations in Frogs.— Guthrie and Guthrie, 1914, Soc. Exp. Biol. Med., n,
144.
Seasonal Changes. — Vanderkleed, 1912, Amer. Jour. Pharm., 84, 14; Central Nervous
System, Donaldson, 1911, ref., Zbl. Bioch. Bioph., 12, 599.
Identification of Frogs by Spots. — Hatcher, 1909, Amer. Jour. Pharm., 23, 303.
Probability Curve. — Tigerstedt, 3.5, 36; Mathematical Methods in Biology, Abderhal-
den, 8, 573.
Experiment 2. (Optional) Focke's Method. — This is based on the acute cardiac death.
It i< ojH-n to the erituism that the time of observation is too short to insure complete
absorption. The details are described by Focke, Zs. exp. Path., 14, 262; Fuehner, Nach-
05: Pittenger, 45.
Experiment 3. (Optional) Guinea-pig Method of Reed and Vanderkleed. — This
determines the minimum hypodermic dose which is fatal in twenty-four hours per 250 gm.
of guinea-pig (for instance, o.i gm. of Digitalis). The details are described in Pittenger,
25. The same method may be used for a number of other drugs, and is official for Aconite
in the U. S. P. IX (see Experiment 6).
>:ical References.— M. F. D. of Cardiac Stimulants and Depressants for Gninea-
pigs. — Githens and Vanderkleed, 1910, Amer. Jour. Pharm., 82, 453. Seasonal Variations,
Vanderkleed, 1912, Amer. Jour. Pharm., 84, 14.
Experiment 4. (Optional) Estimation of Activity on Excised Heart. — The method is
adapted to special research problems rather than to routine assay.
References. — Straub, 1910, Bioch. Zs., 28, 395; Mendenhall, 1915, Jour. Pharmacol., 6;
Krailsheimer and Schmiedeberg, 1910, Arch. exp. Path., 62, 296.
Experiment 5. (Optional) Cat Method of Hatcher. — This determines the M. F. D.
for cats on slow intravenous injection. This "cat-unit" corresponds to o.i mg. of ouabain
per kilogram. In case of slowly acting digitaloids only a partly fatal dose is given, and
the reaction is completed with the ouabain. Details and doses, Hatcher and Brody, 1910,
Amer. Jour. Pharmacy, 82, 362; Jour. Amer. Med. Assoc., 54, 1050; Pit tender, 31.
Experiment 6. (Optional) Gold-fish Method of Pittenger and Vanderkleed. — Jour.
Amer. Pharm. Assoc., 4, 427, 1915.
Experiment 7. (Optional) Bio-assay of Aconite, Official U. S. P. IX Method. — This
< on-i.sts in the determination of the hypodermic dose just fatal to the guinea-pig in twelve
hours. The standard dose per gram of pig is 0.00004 c-c- °f the Fluidextract, 0.0004 °f
Tincture.
Technical References. — Fuehner, Xach weiss, 102; Arch. exp. Path., 66, 179. Other
bio-methods, including Squibb' s Taste Method, are described in detail by Ford, Ford and
Wine, KM 5. Amer. Jour. Phar., 87, 489.
EXERCISE IV.— (ALL GROUPS) EXPOSED FROG HEART
I>ORTER V, C)
Inspection of the exposed heart often reveals certain phenomena, es-
pecially irregularities, more satisfactorily than do tracing. The dru^s may
be applied directly or administered systemically, especially into the 1\ mph-
sac of the thi^h.
Exposure of Heart of Frog.— Pith, decapitate, or anestheti/.e tin- animal by injecting
0.2 gm. "I" t'rrthane (a « .« . of 10 per cent.) into the lymph-sac. In ten t<> fifteen minute-.
paralyzed, raise a fold «f -kin with fon eps. and * ut .m.iy .1 -trip. n<>:
U region. \\ ith ft i— ors divide the « enter «>f tin- -ternum from
above; the lowest « artilagimm^ portion i- . ut -onuvvhat to the left to avoid the median
The arms are pulled apart and fixed to a -mall hoard with pin-. Tin- heart • an
then be seen 1>« to be treated with reagent-, the peruardium should be
opened. The frog's heart will be seen t.. consist of t\\<> .uiri. le- ami a -ingle ventri.le.
mall, whitish hulhus aorUe, and from this the two .\
If the he.ir 1 up, it will he -een that the auricles are lontinued into the sinus
\ \\hitt- line m.irk^ the jun. lion of the two. The stimulation of this line
-timulate /.mirlia. If the heart i- to l>e handled considerably, it will he . on
pbcc .1 -ilk ligature around the frenum, the delicate fibrous band at ta. lung the
lower surface of the heart to the pericardium. This can then be divided and the heart
turned by the ligature.
I96
A LABORATORY GUIDE IN PHARMACOLOGY
Injection of Drugs. — The drugs are injected into the lymph-sac of the left thigh (which
is not fastened), inserting the needle about i cm. below the knee-joint of the extended leg,
and pushing it upward. After the injection is made the knee is flexed to prevent leakage.
Local Application. — Solutions may be applied directly to the heart with a pipet or
camel's hair brush. The application should be renewed every five minutes, just after the
observations.
Observations. — These should bear on the rate of auricles and ventricle; the size, rela-
tive strength, and duration of systole and diastole, the color and regularity. The results
should be plotted as curves, as shown in Fig. 38.
Tracings. — These may be taken either by (a) resting a light lever directly on the heart;
or (6) by attaching a small piece of cork to the muscle-lever, in place of the weight, and
resting this weight on the heart; or (c), by the suspension method, passing a fine thread
around or through the apex, and connecting with a muscle-lever; or (d) by connecting
one of the aortae with a small mercury manometer. When levers are used with the heart,
they should be light and well balanced.
Technical References. — Fuehner, 90; Heinz, i, 820; Robert, Intox., i, 193; Greene, 69;
Tigerstedt, 2.4, 123.
Ail nates after
Fig. 38. — Diagram of observations on the effect of digitalis, frog's heart.
Experiment i. (Group I) Digitalis, Inspection. — Anesthetize frog with
Urethane. After ten minutes inject into thigh lymph-sac Tr. Digitalis,
i c.c. After five minutes expose the heart and continue the observations
until the heart stops. Plot -curves as shown in Fig. 38. The effects consist
in an increased tone of the cardiac muscle; the beats are slowed (sometimes
there is a preliminary quickening) and strengthened. The systole particu-
larly increases, the heart becoming progressively smaller and whiter. The
contractions then become irregular and often peristaltic. The slowing
continues and affects particularly the ventricle, so that there may be several
auricular beats in each contraction of the ventricle. Finally the heart
stops in systole, i. e., as a small white lump. It may be necessary to apply a
20 per cent, infusion to obtain this result. If the ventricle be distended
by injecting 0.75 saline under pressure (with a hypodermic syringe), it
will again contract. The application of aqueous camphor solution, or
pricking with a needle, starts only a few beats.
(Optional) Local Application. — Instead of injecting the digitalis, it may be applied
locally as 5 per cent, infusion. Veratrin (2 per cent.) or BaCl2 (i per cent.) give effects
very similar to digitalis.
The results are sometimes atypical.
CHAP. XXXVI EXCISED AND FROG HEARTS IQ7
Experiment 2. (Group II) Digitalis Tracing. — Inject Urethane and Digi-
talis as in Experiment i ; but after exposing the heart, insert a hook in the
apex, connect with a heart-lever, and take slow-speed tracings.
(Optional) Ouabain. — In place of digitalis, in the above experiments, inject ouabain,
o.oi mg. (\ c.c. of i : 50,000).
Experiment 3. (Group III) Aconite Tracing. — Anesthetize frog, expose
lu-art, and take normal tracing. Inject into thigh 20 mg. of aconite (0.5 c.c.
of 4 per cent.): successively, increase of rate, cardiac peristalsis, diastolic
arrest .
Experiment 4. (Group IV) Aconite, Inspection. — Pith the brain of a frog,
explore the heart, open the pericardium, and apply a 4 per cent, infusion
•nite. Plot curves as shown in Fig. 38.
Aconite stimulates and then paralyzes successively the accelerators,
vagus, and muscle. If the results are typical the rate is first quickened, then
cd, then again quickened and irregular, and then gradually slowed, with
final paralysis in the median position. The primary quickening may be
absent. The secondary quickening is fairly constant and characteristic.
The most striking feature is the extreme irregularity and arhythmia of the
heart in the later stages. This may take the most varying forms. The
t\v<> sides of the ventricle often beat alternately, the blood being pumped
from one side to the other.
Experiment 5. (Group V) Comparative Toxicity of Anesthetics. — Excise
the hearts of three frogs. Place a heart in each of three watch-glasses
containing the following normal solutions: (a) Normal saline; (b} normal
saturated with chloroform; (c) normal saline saturated with ether,
that the chloroform stops very quickly, the ether heart much later.
The stoppage is in the median (paralytic condition), and is preceded by slow-
ing and weakening. If the hearts are at once removed to normal saline,
they may beat again.
The greater toxicity of the chloroform is emphasized by the fact that it is
much less soluble than ether, the saturated solution containing only a
: icth as much of chloroform as of ether.
Questions. — (a) Describe the effects of digitalis, aconite, and anesthetic-.
(b) Is ether or chloroform more dangerous?
EXERCISE V.— OPTIONAL EXPERIMENTS ON FROG HEART
Experiment i. Chloral and Camphor.— Injn t into ventral lymph--. 40 mg.
!-»r.il '0.4 < ,C. <>f 10 i>er cent.). In ten or fifteen minute> e\p«i>e the heart ami
tra«inn. When heart a. lion U \\eal. and slow, irrigate with N Mtmli. and then
with saturated solution of camphor in \. S. The heat is materially Mren^thened iHiihme.
1905, Arch. exp. Path. I'harm., 52, 347). The >timulati<m is >een only on depressed frog
hearts and not in mammaU Plant, 1^05. Jour. Pharma«ol
Experiment 2. Other Drugs for Lymph-sac Injections and Tracings.— Ahohol. i
lit.; Morphin. 10 per
'>stiKmin. o.i «
Experiment 3. Other Drugg for Local Application and Inspection. Antipyrin. i JMT
Inral. i |" 'iloroform. 0.5 per »ent.: Pot.i
ntiinin. o.i p< t . hnin. o.oi per
Experiment 4. Lymph Hearts of Frog.— Kohert, Int«\ . i. 195.
198 A LABORATORY GUIDE IN PHARMACOLOGY
EXERCISE VI.— (ALL GROUPS) PERFUSION OF TURTLE HEART
(REPORTER I, D)
Technic.— Arrange a Mariotte perfusion bottle (Fig. 39) with about 250
c.c. of Ringer's Fluid, connected with a rubber tube about 25 cm. long,
furnished with a pinch-cock and ending in a cannula of about 2 to 4 mm. end
diameter, for insertion into the vena cava. Fill the connections with the
fluid, have ready another 25-cm. rubber tube, ending in a cannula of i to 3
mm. end diameter, for the aorta. The other end of this tube is furnished
with a bent glass tube. Fasten the bottle on a stand about 20 cm. above
the table.
Draw out the head of the turtle and destroy the brain by a blow with a
hammer. Cut through the junction of the lower shell (plastron) with a
saw or bone forceps and remove it with a scalpel. Expose the heart and
remove the pericardium. The animal can
be supported on its back by a towel twisted
into a ring. Insert the cannulae into a vena
cava and into the aorta and tie all other
vessels. Excise the heart.1
Connect with the perfusion apparatus,
avoiding air-bubbles. Fix the cannulae in
a clamp so as to support the heart firmly.
Place a hook or clamp on the apex of the
heart and connect with a lever tracing on
slow drum. The level of the perfusion
fluid should be about 10 cm. above the
heart. Place the free end of the aortic
tube in a graduate, about 15 cm. above the
heart, and measure the outflow per minute
or other convenient period.
Experiment i. (Group I) Antipyrin and
Epinephrin. — Obtain normal tracing and
observations. Add Antipyrin2 to the per-
fusion bottle in the proportion of i : 4000
(2.5 c.c. of i per cent, per 100 c.c. of
Ringer's). When the contractions have
become very weak, inject slowly with a
hypodermic syringe into the vein-tube
about i c.c. Epinephrin, i : 100,000: stimu-
lation.
Experiment 2. (Group II) Aconite and Epinephrin. — Proceed as in Ex-
periment i, using Aconite, i : 500 (2 c.c. of Tincture per 100 c.c. of Ringer's).
The heart passes through the peristalsis to a final slowing. Inject Epinephrin
as in Experiment i : stimulation.
Experiment 3. (Group III) Alcohol and Epinephrin. — Obtain normal
tracing and observations. Add Alcohol to the perfusion bottle, raising the
concentration (with observations and tracings), progressively, through f , i,
and 5 per cent. (J, i, and 5 c.c. per 100 c.c. of Ringer's Fluid): the lower
concentration is inactive, the higher produces some depression. Inject
Epinephrin as in Experiment i.
1 The heart may be left in position. This is more convenient, but becomes disturbing if the
animal should move.
2 Or Phenol, i : 5000 (2 c.c. of i per cent, per 100 c.c. of Ringer's).
Tig. 39. — Perfusion of turtle heart.
CHAP. XXXVI EXCISED AND FROG HEARTS 199
(Optional) P/iysosti^min, 0.5 c.c. of i per cent, per 100 c.c. may be used
in place of Epinephrin.
Experiment 4. (Group IV) Potassium and Epinephrin. — Proceed as in
Experiment i, using Potassium Chlorid (5 c.c. of 10 per cent, per 100 c.c. of
Ringer's Fluid). When the heart is greatly weakened or arrested, inject
Epinephrin as in Experiment i.
Experiment 5. (Group V) Digitalis and Potassium. — Proceed as in Ex-
periment i, using Digitalis i : 10,000 (o.i c.c. of Tincture per 100 c.c. of
Ringer's). When the heart has gone into systolic standstill, see whether
it can he started by raising the pressure in the aortic tube.
Inject into the vein tube i c.c. of 10 per cent. KC1. If this does not
start the heart, see whether it can be recovered by perfusion with unpoisoned
Ringer's Solution.
Questions. — (a) Describe the effects of the drugs.
(b) Is the digitalis standstill due to paralysis of the cardiac muscle?
Why?
EXERCISE VII.— (DEMONSTRATION) VAGUS POISONS ON TURTLE
(REPORTER III, D)
Technic. — Destroy the brain of a turtle and remove the plastron, as
explained in Exercise VI. Draw out the head, so as to put the neck on the
stretch and fasten it in position by a nail. Cut away skin and fascia at
base of neck and dissect the vagus nerves: they emerge from the long
retractor muscles of the head just posterior to where the coracohyoid
muscles join in the median line. In the upper part of its course the \
lies internal to the retractors, then winds around to the front of these
muscles (Edmunds and Cushny, 139). It is accompanied by the sympa-
thetic, from which it can be distinguished by the results of electric stimula-
tion. (The right vagus is much more effective, Carrey, 1911, Amer. Jour.
Physiol., 28, 330.) The electrodes may be left in place. Attach a hook to
the apex of the heart, attach to a lever, and take normal tracing. (A special
turtle myocardiograph is described by Cushny, 1905, Arch. Intern. Pharma-
cod, 15, 493; Edmunds and Cushny, Lab'y Guide, p. 144.) Make the fol-
lowing experiments, taking tracing:
1 . Record effect of stimulation of vagus.
2. Paint heart with 0.5 per cent. Pilocarpin. When contractions have
become very slow, paint with
3. Atropin. o.i per cent.
4. Note that stimulation of vagus is now ineffective.
5. Again paint with Pilocarpin. and note that response to vagus returns
more or less perfectly.
i>e the heart, place in 0.75 XaCl, and keep for Exercise IX.
QUESTIONS
(a) Describe the effect of pilocarpin.
(b) How i> tin- atTected by atropin?
(c) What rtYect has atropin on the vagus?
' light does thi- throw {m the mechanism of the pilot
slowing?
2OO
A LABORATORY GUIDE IN PHARMACOLOGY
EXERCISE VIII.— (DEMONSTRATION) VAGUS POISONS ON FROGS
(REPORTER III, D)
The same experiments can be performed as in Exercise VII, but frogs
are less satisfactory, because the vagus trunks sometimes do not respond to
the stimulation. Frogs, however, are well suited for studying vagus ganglia.
These are reached by lifting the heart and stimulating the junction of the
auricles and sinus venosus.
Experiment i. — (a) Pith the brain of a frog, pin on board, expose heart,
and remove pericardium. Note that electric stimulation of the sinus venosus
stops the heart (stimulation of vagus ganglia).
(b) Apply atropin (i : 1000): In a few minutes stimulation of the sinus
produces no effect (paralysis of vagus endings). The atropin may cause a
quickening of the heart by stimulating the muscle.
(c) Wash off the atropin with normal saline. Apply muscarin (i : 1000)
(or physostigmin) : sinus stimulation is again effectual, and heart may be
slowed (stimulation of vagus endings and cardiac muscle).
(d) Wash with normal saline and repeat (b) : same effect. Atropin and
muscarin (or physostigmin) have antagonistic actions, and whichever is used
in larger quantities can overcome the effects of the other. This holds for all
peripheral structures upon which these alkaloids act.
Experiment 2. (Optional) Quantitative Estimation of Muscarin by Excised Heart. —
See Fuehner, 1908, Arch. exp. Path. Pharm., 59, 179 (Nachweis, 137).
Technical Notes on Cardiac Nerves of Frog. — The vagus trunk comes to the surface
at about the angle of the jaw, in company with the glossopharyngeal and hypoglossal
Fig. 40. — Dissection of vagus, frog: v, Vagus nerve; h, hypoglossal nerve; g, glossopharyngeal
nerve; b, brachial plexus; j, jaw.
nerves, lying between the two. By exposing this area the vagus can easily be seen pass-
ing to the heart (Fig. 40). It may be dissected out and placed on a ligature for stimula-
tion, but frequently it suffices to stimulate it in situ.
For the dissection of the accelerator nerve, see Stewart's Manual.
QUESTIONS
(a) Does atropin paralyze the vagus ganglia?
(b) Where, then, must its action be situated?
CHAP. XXXVI i \< IM 1) AND FROG HEARTS 2OI
(c) Since muscarin or physostigmin act after atropin, where could their
action be situated?
(d) Since atropin also acts after these, where must their actions be
located?
EXERCISE IX.— (ALL GROUPS) DRUGS ON STRIPS OF TURTLE'S
VENTRICLE
(REPORTER I, D)
Use the ventricle of the turtle used in Exercise VII. Grasp the left
angle of the base of the ventricle with forceps and cut around the apex to
the opposite side. This piece may be cut into two or three strips and
attached to a hea^vy muscle lever, weighted with i ,mn., precisely like a gas-
trocnemius preparation, keeping it immersed in 0.75 per cent. XaCl. Con-
tractions begin in ten to forty minutes. Take normal tracings, and add the
following drugs:1
Experiment i. (Group I) Alcohol, successively 2, 5, and 10 per cent.
{0.4, i, and 2 c.c. per 20 c.c. N. S.).
Experiment 2. (Group II) Strychnin, i : 10,000 (2 c.c. of i : 1000 per
20 c.c. N. S.); after ten minutes, caffcin, i : 1000 (2 c.c. of i : iooper 20 c.c.
X. S
Experiment 3. (Group III) Ouabain, i : 100,000 (2 c.c. of i : 10,000 per
20 c.c. X. S.): digitalis action.
Experiment 4. (Group IV) Potassium Chlorid, i : 200 (i c.c. of 10 per
cent, per 20 c.c. X. S.). When heart is weakened, add Epincphrin, i : 20,000
(i c.c. of o.i per cent, per 20 c.c. N. S.).
Experiment 5. (Group V) Calcium Chlorid, i : 200 (i c.c. of 10 per cent,
per 20 c.c. X. S.). When heart is weakened, add Epincphrin, i : 20,000(1
c.c. of o.i per cent, per 20 c.c. X. S.).
QUESTION
Describe the effects of the drugs.
TECHNICAL REFERENCES
«cne, 66.
ilniRx illicit may be used arc: Atropin. o.ooi and 0.002 per cent.; Barium
Chlorid, o.oi percent.; Chloral, o.o'i percent.; Chloroform. 0.05 and o.i per 06B1
o.i per cent.: Digitalis, 0.002 and 0.005 P^r ct>1 ' ° !K'r ''i'»t.; Kthcr. i. :, 4. and
6 permit.: Morphin, i JHT . ml.; N'i.otin. 0.05 percent.; Nitrite of Sodium, o.o: per
:iin, o.i IKT cent.: I'ilo( arpin, o.i prr trnt.: Ycratrin. 0.005 and 0.05 percent.
•.'mali Heart of Chick Embryos. <>r twenty-four to thirty -i\
hours, larcfully opened, the contents llo.ited in a dish, and the membranes cut .1
The heart-beat may be observed in a wat. h ula— . under tl >c,and drop applied,
fhe heart at thi- time d«>r- n«»t . ..ntain nerves. (Pickering* 18930
1 The preparation, after a normal tracing has been taken, may be immersed in the drug until
the effect starts, and then returned to the unpoisoned
202
A LABORATORY GUIDE IN PHARMACOLOGY
CHAPTER XXXVII
AUTONOMIC DRUGS: (A) PUPILS; (B) GLANDS; (C) BRONCHIOLES;
(D) ANAPHYLAXIS; (E) EXUDATIVE INFLAMMATION
(A) EFFECTS OF DRUGS ON THE PUPIL
Introduction. — The iris contains two sets of smooth muscle-fibers, the circular sphinc-
ters, and radial dilators (Fig. 41).
The sphincter muscle is innervated by fibers contained in the oculomotor nerve.
These terminate around the cells of the ciliary ganglion. From here the fibers pass on
as the short ciliary nerve.
Fig. 41.— Innervation of iris (adapted from P. Schultz): Solid line = sympathetic (dilator); fine
dotted line = oculomotor (constrictor) ; coarse dotted line = trigeminal.
The nerve-fibers for the radial muscles run in the cervical sympathetic nerve, and
terminate in the superior cervical ganglion. The fibers which arise here run through the
Gasserian ganglion (but without joining any cells), where they unite with the first branch
of the trigeminal, and run to the iris in the long ciliary nerve.
The pupils may, therefore, be affected through the following mechanisms:
(A) DILATOR MECHANISM.
1. Sympathetic center.
2. Sympathetic and long ciliary nerve.
3. Superior cervical ganglion.
4. Postganglionic fibers.
5. Endings in radial muscle.
6. Fibers of radial muscle.
CONSTRICTOR MECHANISM.
7. Oculomotor center.
8. Oculomotor and short ciliary nerves.
9. Ciliary ganglion.
10. Postganglionic fibers.
11. Endings in sphincter muscle.
12. Fibers of sphincter muscle.
Stimulation of "A" causes dilatation; paralysis, constriction through the unopposed
action of the constrictor mechanism.
Stimulation of "B" causes constriction; paralysis, dilatation through the unopposed
action of the dilator mechanism.
The action may be located as follows (the principal drugs giving these effects are added
in parentheses):
A. It is tried whether the drug acts also when applied to the cornea, and if so, whether
the effect is confined to this eye, or at least is much greater there. If this is the case, the
action must be on the endings or muscle. If the drug acts only when it is introduced sys-
temically, the action must be on the ganglia or centers. The ganglia are discussed below,
Central actions are usually confined to the dilator center (stimulated by asphyxia, depressed
in man by morphin).
B. Dilation of Pupil (Mydriasis). — The oculomotor trunk is exposed and stimulated:
1 . No effect. Peripheral constrictor paralysis. It remains to distinguish between the
ganglia, endings, and muscle, by stimulation of the short ciliary and of the sphincter
muscle. (Atropin paralyzes the oculomotor endings. What would be the result of these
stimulations?)
2. Oculomotor stimulation is effective. The dilation must be due to sympathetic
stimulation. The drug would be ineffective after section and degeneration of the sympa-
CHAP. XXXVII AUTONOMIC DRUGS 203
thetic. Stimulation of the ganglia can be shown or excluded by section of the long ciliary.
(Cocain stimulates the sympathetic center, ganglion, and endings. Epinephrin stimu-
lates the myoneural function.)
C. Constriction of the Pupil (Miosis). — The cervical sympathetic is stimulated:
1. \o effect. Sympathetic paralysis. The distinction between ganglia, endings, and
muscle is made by stimulating the long ciliary and the radial muscle. (Nicotin paraly/.c.-
thc ganglia after a preliminary stimulation.)
2. Sympathetic stimulation is effective. The constriction must be due to oculomotor
stimulation. This is generally in the endings (physostigmin, muscarin, pilocarpin). The
ganglia may be excluded by section of the short ciliary; the muscle by the fact that large
doses of atropin cause dilation
The localization of these actions requires rather complicated operations; but the local
effects and the antagonism can be readily demonstrated.
TECHNICAL REFERENCES ON SPECIAL SENSES
Pupils. — Robert, Intox., i, 212, 281; Fuehner, 144.
Subcorneal Inoculation. — Abderhalden, 3, 1285.
Cataract. — Robert, Intox., i, 215.
Chcmosis. — Ibid., i. _M5.
Iritis and Uveitis. — Guillery, 1915 (prodigiosus ferment), Zentr. Bioch. Bioph., 18, 71.
Light Sensation. — Tigerstedt, 3.2, i ; Color sensation, ibid., 42; Eye movements, ibid., 100.
Ophthalmoscopy— Tigerstedt, 3.3, 55.
Cranial Nerves, Operations. — Tigerstedt, 3.4, 101.
Special Senses. — Temperature, Tigerstedt, 3.1, i; Pressure, ibid., n; Pain, ibid., 30;
Odor, ibid., 46; Taste, ibid., 91.
Ear. — Innervation, Tigerstedt, 3.3, 181; Hearing, ibid., 204; Acoustics, ibid., 204.
EXERCISE I.— (DEMONSTRATION) LOCALIZATION OF ATROPIN ACTION
ON PUPIL1
(REPORTER V, B)
Technic. — Anesthetize dog. Tracheotomize. Divide both vanosympathetus and
arrange central end for stimulation (right side). Turn right side of heart upward. Make a
J.-shaped incision through skin, the vertical limb from sagittal suture to external canthus;
Fig. 42.— Pupillary nerves.
the horizontal limb from internal ranthus of right eye along upper border of orhil and
wliole lower l)order of /yu'uma. Stop all hemorrhage. C'ut away the upper , art ilairim>u>.
orbital border. Open the orl.ital < ap-ulc below the external rr.tu-. Divide and relle, t
the 1 •••lullx , 1,-aii a\\ay tatty tifltUC until optic net I >raw billing
forw. .rch for plan'- where short « iliarics leave the optic sheath. or
search direi tly for the « iliary ganglion. by drawing tlu- inferior rectus muscle outuard and
tra. tOTDUlbi u|«w;»r.: under llu- ^hort • iliary and long » iliary nerves.
(The IOM^ , iliary nerves also run on the npti< n« •
Confirm the detection by efectlil vtinml.ition the >h..rt cOUfiet OOOStlkl the pupil.
tlie l.,ni: , iliari-- If there is any dim, ulty in 1... itter. stimulati
:ral \ami^ may lie >ul»st it uted.
Experiments.— Ha\ in- (.mtirnn-d tin- effects of stimulation, in
!roj)s«»f Atroj)in (i : 1000) into tin- anterior chamlKT: tin- pupil dilates.
1 Jegorow, Arch i S86, 1 50.
204 A LABORATORY GUIDE IN PHARMACOLOGY
Stimulation of the short ciliary is now ineffective, stimulation of the long
ciliary still causes dilation. To show that the sphincter fibers themselves
are not paralyzed separate the points of the electrodes to the diameter of
the pupil, and thus stimulate, giving a circular motion to the electrodes:
the pupil constricts. If the animal is in good condition, inject a few drops
of Physostigmin (i : 1000), and see whether the excitability of the oculo-
motor is restored.
QUESTION
Give the evidence showing that atropin paralyzes the oculomotor
endings.
EXERCISE H.— (ALL A GROUPS) LOCAL APPLICATION OF MYDRIATICS
AND MIOTICS TO MAMMALIAN EYE
(REPORTER V, B)
General Method. — Drop a few drops of the solution into the eye of the
animal with a pipet. Note when the dilatation or constriction sets in —
about fifteen minutes (using the other eye for comparison) ; when it reaches
its maximum — about an hour; and when it disappears — about a day. Try
whether the light-reflex is preserved. Report the results, stating what con-
clusions are justified in each case. Cats are best adapted to the study
of drugs acting on the pupil. Dogs answer very well. Rabbits can also
be used, but are not quite as sensitive. It must also be remembered that
in rabbits the two eys react independently to light, so that the nose of the
rabbit must be pointed to 'the window if the eyes are to be compared.
Rabbits do not react to Dionin.
Experiment i. (Group I, A) Atropin, Pilocarpin, Physostigmin. — (a)
Place 2 drops of Atropin (i : 1000) into the eye of the animal: dilation.
The effect is confined to one eye. Light-reflex is absent. (Paralysis of
oculomotor endings.)
(b) In an hour drop Pilocarpin (i : 100) into the same eye: little effect.
(c) In fifteen minutes drop Physostigmin (i : 100) into the same eye:
constriction.
Experiment 2. (Group II, A) Physostigmin.— Into the eye of another
animal place 2 drops of Physostigmin (i : 100): constriction confined to the
one eye. Appears in fifteen minutes, maximum in about an hour. (Stimu-
lation of oculomotor endings.)
Experiment 3. (Group III, A) Pilocarpin.— Drop Pilocarpin (i : 100) into
the eye of another animal : constriction confined to the one eye, but not as
great as Physostigmin. (Peripheral stimulation of the oculomotor.)
Experiment 4. (Group IV, A) Cocain. — Drop i : 100 solution into eye of
another animal. Note the anesthesia and dilatation confined to the one
eye. The latter is not as strong as with Atropin, and the pupils still react
to light. (Stimulation of the sympathetic.)
Experiment 5. (Group V, A) Dionin (Ethylmorphin) . — Drop some 10
per. cent, solution on conjunctiva of dog or cat: hyperemia and edema.
QUESTIONS
(a) Which of the drugs are mydriatics?
(b) Which are miotics?
(c) Which is more powerful, atropin or cocain?
(d) Which is more powerful, pilocarpin or physostigmin?
(e) Give evidence showing that the actions are peripheral.
(/) Describe the effects of dionin.
CHAP. XXXVII AITOXOMIC DRUGS 205
EXERCISE HI.— (ALL B GROUPS) FROG PUPIL
(REPORTER V, B)
Frogs are not generally quite as satisfactory as mammals for the study of
pupil changes, but they illustrate some interesting phenomena of antago-
nism. They are also much more subject to epinephrin and may be used in
testing for this drug.
Technic of Excised Eyes. — Pith frog. Insert scissors in mouth and
cut off head, except lower jaws. Cut away lower lids. Cut head in two,
lengthwise. Pin each piece on cork, cornea straight upward. Cut rings
about 3 mm. high from rubber tubing of about same diameter as eye.
Place a ring on each eye. This forms a little cup, into which the solution is
dropped (Meltzer, 1909, Deut. med. Woch., Xo. 131).
Experiment i. (Group I, B) Pilocarpin and Physostigmin. — Prepare the
eyes as described. Leave in dark until pupils are dilated. Then apply to
one a few drops of i per cent. Pilocarpin ; to the other i per cent. Physos-
tigmin: both are constricted.
Experiment 2. (Group II, B) Atropin, Pilocarpin, Physostigmin. — Pre-
pare the eyes as described. Apply to both a few drops of i per cent. Atropin.
When pupils have dilated, wash and apply to one i per cent. Pilocarpin, to
the other i per cent. Physostigmin. The pupil of the Physostigmin eye
constricts, Pilocarpin does not.
Experiment 3. (Group III, B) Nicotin or Curare, Pilocarpin, Physostig-
min.— Prepare both eyes as described. Apply to both o.i per cent, solution
of Nicotin (or i per cent, curare) and leave in the dark. After half an hour
wash and apply to one i per cent. Pilocarpin, to the other i per cent.
Physostigmin. Pilocarpin constricts, Physostigmin very little (Dixon &
Maiden, 1908, Jour. Physiol., 37, 531).
Experiment 4. (Group IV, B) Cocain, Pilocarpin, Physostigmin. — Pre-
pare both eyes as described. Apply to both i per cent. Cocain: the pupils
dilate. Wash and apply to one i per cent. Pilocarpin, to the other i per
cent. Physostigmin: both constrict.
Experiment 5. (Group V, B) Epinephrin. — (a) Prepare the eyes as
described. Apply to both Epinephrin, i : 10,000: submaximal dilation.
Wash and apply to one Pilocarpin, i per cent., to the other Physostigmin,
i per cent.: both constrict. (Ehrmann, 1905, Arch. exp. Path. Pharm..
53 : 97 (Fuehner, 146) tried to elaborate thi> reaction into a quantitative-
method, but Metzer, loc. cit., finds it unsuited to this purpose.)
(b) Pith a frog and inject into lymph-sac o.oi mg. of Epinephrin (i c.c.
100,000): dilation of pupil.
Furtln-r characteristics of the Epinephrin mydria>i> are that it is sub-
maximal: tin- pupil- become round; they do not react to light (Metzer,
loc. i
QUESTIONS
(a) Which of the drills are mydriatics?
(b) Which arc- minti.
i date tin- eit'u ienrv of pilooarpin and of physostigmin
atropin. nic nt in or curare, cocain. and epinephrin.
(d) Assuming that atropin paraly/.r- the nrul. .motor ending, wlu-rc
'ii oi piloearpin and of ph\ IK- !<>< at
(e) How b this limited by the fed that atropin di; physostiizmin?
(/) Can the mechani-m of the- mydria-i- 1>\ nic <> tin, cocain, and <pi-
nephrin be the same as that of atropin? Why?
206 A LABORATORY GUIDE IN PHARMACOLOGY
(B) EFFECTS OF DRUGS ON (SALIVARY) GLANDS
Introduction. — The peripheral effects of drugs on the iris, on other forms of unstriped
muscle, on the vagus mechanism of the heart, and on glands are very similar. The only
important exceptions are the muscle of the arterioles and uterus, and the liver, mammary
gland, and kidney. The action is the more typical, the more the organ is normally under
nervous control.
The more important drugs act as follows:
Pure paralysis of endings: Atropin.
Stimulation of endings: Physostigmin, muscarin, pilocarpin.
Ganglia: These are first somewhat stimulated, and then depressed, by nicotin, coniin,
lobelia, spartein, curare, and cocain.
The glandular effects are studied most conveniently on the salivary glands. The
submaxillary gland of the dog has the additional advantage that it possesses a double
nerve supply. This may be utilized to prove that atropin acts on the endings and not on
the gland cells.
Glandular secretion may also be affected through the centers, directly or reflexly.
The salivation during apomorphin nausea is an instance of direct central stimulation.
EXERCISE IV.— (OPTIONAL) CHORDA TYMPANI EXPERIMENT
See Stewart's Manual, 450, or Practical Physiology, Beddard, etc., for technic. Insert
a cannula in Wharton's duct. Stimulate the cervical sympathetic: the gland becomes
pale and secretes a little thick saliva. Stimulate the chorda tympani: the gland flushes
and yields abundant thin saliva. Inject intravenously 30 to 40 mg. of nicotin for a dog,
or 10 mg. for a cat. Stimulation of the chorda is now ineffective, but stimulation at the
hilus of the gland (i. e., beyond the ganglion cells) causes secretion. The nicotin has
therefore paralyzed the ganglion. Inject 10 to 14 mg. of atropin for a dog, 5 to 15 mg. for
a cat. Stimulation at the hilus causes no secretion, although the gland flushes. The
atropin therefore does not act on the vasodilator endings, but it paralyzes the secretory
mechanism somewhere peripheral to the ganglion. Stimulate the sympathetic: this
causes secretion. The cells are therefore not paralyzed. The atropin must act on the
endings. Inject some 2 per cent, pilocarpin into the duct, so that it comes in contact with
the cells: secretion resumes, since the pilocarpin stimulation overcomes the atropin paral-
ysis.
QUESTION
State the evidence for the localization of the actions of nicotin, atropin,
and pilocarpin.
TECHNICAL REFERENCES
Circulation of Submaxillary. — Tigerstedt, 2.2, 159.
Distinction of Preganglionic and Postganglionic Fibers (Langendorff, 1892, Cbl. Physiol.,
5, 130). — Stimulation of preganglionic becomes ineffective soon after death; the post-
ganglionic remain excitable for one-quarter to three-quarters of an hour (Langley, 1893,
Jour. Physiol., 15, 181).
Experiments on Saliva. — Robert, Intox., i, 245; Examination, Abderhalden, 3, 257.
EXERCISE V.— (DEMONSTRATION) PILOCARPIN AND ATROPIN ON IN-
TACT ANIMALS
(REPORTER II, A)
The effects may be studied on intact cats or rabbits as follows:
If cats are used, the changes in pulse-rate should also be recorded.
If rabbits are used, the peristalsis may be watched through the abdom-
inal wall.
Experiment i. Pilocarpin. — Inject hypodermically into rabbit or cat
Pilocarpin, 5 mg. per kg. (0.5 c.c. of i per cent, per kg.): intense salivation
occurs in about half an hour. Peristalsis is greatly increased (diarrhea). The
pulse is first slowed, then quickened. The pupils may be constricted. Keep
the animal as control for Experiment 2.
Experiment 2. Pilocarpin Antagonized by Atropin. — Inject another
animal as in Experiment i. When salivation is marked, inject Atropin,
CHAP. XXXVII AUTONOMIC DRUGS 207
10 mg. per kg. (i c.c. of i per cent, per kg.). On comparing the animals
after about half an hour, it will be seen that the Atropin has checked the
salivation and peristalsis, quickened the pulse, and dilated the pupils.
Excitement and forced movements may accur.
QUESTIONS
(a) Describe the effects of pilocarpin on saliva, peristalsis, heart-rate,
and pupils.
(b) Which of these effects are antagonized by atropin?
TECHNICAL REFERENCES
Quantitative Antagonism of Pilocarpin and Atropin, Cushny, 1915, Jour. Pharmacol.,
6, 439-
EXERCISE VI.— (ALL STUDENTS) REFLEX SECRETION OF SALIVA
(REPORTER II, A)
Place a little dilute acetic acid in the mouth and note the increased
salivation. The inhalation of ether acts in the same manner.
TECHNICAL REFERENCES
Pulse-rate of Mammals (A. Reichert, 1909, Bioch. Cbl., 10, 170):
Horse 30-40 Large dog. 7 2- 82
Cow ....... 70-85 Small dog. 70- 90
Ox. 52-68 Cat.. 116-128
70-86 Rabbit . i .-0-140
Goat 70-90 Chicken 180-200
Bronchial Secretion. — Henderson and Taylor, 1910, Jour. Pharmacol., 2, 153; J. L.
Miller. 1014. Amer. Jour. Med. Sci., 148, 469.
Mm us, l''ro% Skin. — Robert, Intox., i, 189.
Sweat. — Robert, Intox., i, 268; Collection, human, Abderhalden, 3, 998, 1000.
Milk. — Robert, Intox., i, 272.
Secretin: Preparation and Tests. — Abderhalden, 3, 205, 418; 6, 487; 7, 65; Dale and
Laidlaw, 1912, Jour. Physiol., 44, XI.
Gastrin. — Preparation, Keetom and Roch, 1915, Amer. Jour. Physiol., 36, 353.
(C) EFFECTS OF DRUGS ON BRONCHIOLES
Introduction. — The bronchial muscles are affected by the autonomic
poisons in the usual manner. For instance, they arc constricted by physos-
tiumin, pilot arpin, and muscarin (stimulation of constrictor endings), and
by barium and histamin (direct stimulation of muscle). They are relaxed
by atropin (paralysis of constrictor endings) and by epinephrin and hordenin
;uilation of dilator endings). Violent constriction occurs in anaphylaxis
and in a>thma. This may l»e treated by atropin or epinephrin.
EXERCISE VII.— (DEMONSTRATION) BRONCHIAL TONE IN LIVING
ANIMALS
i i i II, A)
I hi- may be (Miniated by the variation of intrapleural pressure, \\ith
taut respiration.
tlu-tize a rabbit. Connect trachea for artificial respiration, inter-
posing an opni T-pieee for free escape of excess of air. The respiration
HUM U uniform in rate and volume. Cut through cervical cord and
destroy respiratory center. Connect jugular vein for injection. Through
208 A LABORATORY GUIDE IN PHARMACOLOGY
a flanged cannula connect one pleura with tambour and take slow tracing
of pulmonary excursions. An increase of these excursions is due to dilation
of the bronchial muscles, and vice versa.
Inject the following drugs into the jugular vein while taking tracings
(the doses are for average animals) :
1. Epinephrin, o.i mg. (i c.c. of i : 10,000): no effect.
2. Pilocarpifij i.mg. (i c.c. of i :iooo): constriction. During this
constriction inject:
3. Epinephrin, as in (i): relaxation.
4. Pilocarpin, as in (2); during constriction inject:
5. Atropin, 2 mg. (2 c.c. of i : 1000) : relaxation.
6. Histamin, o.i mg. (i c.c. of i : 10,000): constriction.
7. Atropin, as in (5); then Epinephrin, as in (i): no relaxation.
QUESTIONS
(a) Which drugs constrict, and which relax, the bronchi?
(b) What essential difference is there between pilocarpin and histamin?
(c) What light does this throw on the site of their action?
TECHNICAL REFERENCES
Methods involving the same principles are described by Dixon and Brodie, 1903,
Jour. Physiol., 29, 97; Golla and Symes, 1914, Jour. Pharmacol., 5, 92; D. E. Jackson,
ibid., 4, 7, 59; 5, 479.
EXERCISE VIII.— (DEMONSTRATION) TREATMENT OF BRONCHIAL SPASM
IN PERFUSED LUNG
(Method of Baehr and Pick, 1913, Arch. exp. Path. Pharm., 74, 41.)
(REPORTER II, A)
A guinea-pig of about 250 gin. is etherized lightly. Insert a tracheal cannula, con-
nected through a T-tube, with one limb open, with a respiration bellows of uniform action.
Remove sternum. Tie a cannula into pulmonary artery, pointing toward lung. Connect
through a T-piece with two perfusion bottles, one filled with glucose-free Tyrode solution,
the other with i per cent. Witte Peptone in Tyrode. Tie a cannula into the apex of the
ventricle for the outflow of fluid (this may be measured if it is desired to study the vascular
action). It is best t,o leave the whole preparation in the thorax. (The excursions of the
lung can be recorded by a lever.)
Adjust the perfusion bottles about 30 cm. above the lung, and start the
perfusion with Tyrode's fluid. Change to the peptone: the excursions
diminish promptly, the lungs remaining rigidly distended, due to bronchial
spasm. The condition is analogous to anaphylaxis or asthma.
Change to the plain Tyrode fluid, to which 0.05 per cent, atropin (5 c.c.
of i per cent, per 100 c.c.) has been added: the spasm is promptly relieved,
the lungs returning to their normal volume and excursions.
(The following drugs may be used. In place of Peptone: Histamin, i : 100,000; Pituit-
ary, 4 per cent, of the fluid; Pilocarpin or Physostigmin, i : 10,000. In place of Atropin:
Epinephrin, i : 100,000; other drugs are described in the original paper.)
QUESTION
What drugs would be efficient against the spasmodic attacks of asthma?
EXERCISE IX.— (OPTIONAL) REACTIONS OF EXCISED TRACHEAL MUSCLE
(See Trendelenburg, 1912, Arch. exp. Path. Pharm., 69, 106.)
CHAP. XXXVII AUTONOMIC DRUGS 2OQ
(D) ANAPHYLACTIC REACTION
Introduction. — The injection of proteins sensitizes animals toward
subsequent injections of the same protein. The phenomena differ quantita-
tively in different animals. In guinea-pigs the most conspicuous effect is
a bronchial spasm, analogous to that produced by peptone, histamin,
pilocarpin, etc.
TECHNICAL REFERENCES
Methods of Anaphylaxis. — Pfeifer in Abderhalden, 5, 525; Zinsser, Hopkins, and
Ottenberg, p. 182. For shock-sensitization of dog, 5 c.c. horse serum, hypodermic; after
twenty-one days about 5 c.c. by vein (Pearce and Eisenberg, 1910). Dog to egg-white,
Edmunds, 1913, Zs. Immun., 17, 127.
Preparation of Protein Poison (from Egg-albumen). — Vaughan and Wheeler, Jour.
Lab. Clin. Met!., i, 55, 1915.
EXERCISE X.— (DEMONSTRATION) ANAPHYLAXIS IN GUINEA-PIG; PRE-
VENTION OF ANAPHYLACTIC EMPHYSEMA BY ATROPIN
(REPORTER IV, A)
Two guinea-pigs are sensitized two weeks previously by hypodermic
injection of o.i c.c. of horse-serum. On the day of the demonstration one
of the animals (B) receives 3 mg. of atropin hypodermically, at least ten
minutes before the demonstration.
Etherize the atropin pig (B) lightly. Expose the jugular vein. Lighten
the anesthesia. With a syringe inject 2 c.c. of horse serum into the vein.
Tie vein and remove anesthetic.
Do the same to animal (A). In a very few minutes the animal (A)
becomes excited, dyspneic, and dies of asphyxia, usually within five minutes.
Open the thorax and note that the lungs are rigidly distended (Auer and
. 1009, Jour. Amer. Med. Assoc., 53, 6).
The atropin pig shows little or no effect. Kill (with chloroform), open
thorax, and note that lungs are normally collapsed (Auer, 1910, Amer. Jour.
Physiol., 26, 439).
QUESTIONS
(a) Define anaphyluxk
(b) How is it produced?
(c) What is the essential phenomenon in guinea-pigs?
(d) Explain how this is relieved by atropin.
EXERCISE XL— (OPTIONAL) ANAPHYLAXIS IN EXCISED UTERUS
le, IMI !, J«>ur. I'harmacol., 4, 167.)
li/.e ;i young virgin guinea-pig with o. i c.c. of horse serum fourteen days pre-
vinu>ly. Kill by blow on head. M\ -lied blood, and Irt it clot. Cut across
abdomen and perfuse aorta with 500(0 loooc.c. of l.o, kc's Dilution to free uterus of scrum.
lerus to 200 c.c. oxygenated I :i«>n warmed in bath, connect with
levers, and take .slow tr.-i. ing (see Chapter XXXIV, I Ml
1. Add to the solution successively 0.5 c.c. of various foreign non-specific sera — cat,
dog. sheep, ox. etc .: no elTei t .
2. Add horse serum so as to give a con. cnt rat ion of i : 5,000,000 (o. i 10,000
per 200 c.c.) : no cffc Miration to I : 1,000,000 by adding furl IK
filiation.
3. Change the l.o(kc solution, obtain normal tra. ing, and add horse serum i : 1000
(0.2 c.c. of undiluted srrumi: maxim. d CODl
4. A/ •'• the Locke solution, obtain t i.i. in/, and again add o. .• « .« .. tin i
of horM- M rum: n. . response (desenn . the I|MM- e(|iii\alent to .inti
C'l solution, obtain tra. in-, and add .\ ... . of horse serum: tonti
6. Change Locke's solution, obtain tra. ing, and add 4 c.c. of the guinea-pig serum:
contraction.
210 A LABORATORY GUIDE IN PHARMACOLOGY
QUESTIONS
(a) Does the anaphylactic sensibility induced by the injections of a specific serum re-
side in the blood or in the tissues? (Experiment 2.)
(b) Is the reaction of the tissue confined to the antigen? (Experiments i and 2.)
(c) Under what circumstances does desensitization occur? (Experiments 3 and 4.)
(d) Is the desensitization specific to antigen or does it apply to all sera? (Experiments
5 and 6.)
(e) Is the anaphylactic reaction qualitatively or only quantitatively different from the
reaction to ordinary serum? (Experiments 4, 5, and 6.)
(E) EXUDATIVE INFLAMMATIONS
These are somewhat allied to anaphylaxis; at least, local exudates are
among the phenomena of anaphylaxis. These are markedly influenced by
calcium, perhaps because this lessens cell permeability.
Suppuration involves positive chemotaxis and cell necrosis. It is
produced by bacterial and certain vegetable proteins; turpentine oil;
mercurials; croton oil; or 5 to 10 per cent, silver nitrate.
EXERCISE XH.— (DEMONSTRATION) CALCIUM ON DIONIN CHEMOSIS
(REPORTER IV, A)
Inject cat in morning hypodermically with Calcium Lactate, 20 mg. per
kg. (i c.c. of 2 per cent, per kg.). In afternoon drop some 10 per cent.
Dionin in eye: no result. Compare with Exercise II, Experiment 5.
(Chiari and Januschke used a drop of mustard oil. Analgesics also influ-
ence the reaction, Januschke, ref. Jour. Amer. Med. Assoc., 61, 522.)
EXERCISE XIIL— (OPTIONAL) PREVENTION OF PLEURAL EFFUSION BY
CALCIUM
(Chiari and Januschke, 1910, Wien. Klin. Woch., 23, No. 12; 1911, Arch. exp. Pharm.
Path., 65, 122.)
About twenty-four hours before the demonstration inject intravenously into two
lightly etherized dogs sodium iodid, i c.c. of 10 per cent, per kg. One dog (A) serves as
control. The other (B) receives at once, hypodermically, calcium lactate, 2 c.c. of i per
cent, per kg. This dose is repeated in six to twelve hours. In twenty-four hours the dogs,
if not already dead, are killed with chloroform, and thorax is opened: the control dog (A)
shows abundant pleural exudations, sometimes pulmonary edema and hydropericardium
(Boehm, 1876, Arch. exp. Path. Pharm., 5, 329). The calcium dog (B) is dry. (Thio-
sinamin, 0.13 gm. per kg. by vein, may be substituted for the sodium iodid.)
QUESTIONS
(a) What effect has calcium on inflammation?
(b) How may this be explained?
(c) Could calcium be useful in serum rash, etc.?
(d) Suggest why it is of little use in clinical pleuritic effusions.
EXERCISE XIV.— (OPTIONAL) SUSCEPTIBILITY OF CAT'S SKIN TO
CROTON OIL
This is increased by feeding with acid, diminished by Ca (Luithlen, 1911, Wien. Klin.
Woch., No. 20). The effect of the local application of magnesium sulphate and calcium
chlorid could also be tried.
O. Loeb and Loewe, 1916, Ther. Mon., 30, 74, advocate young pigs for experiments
with cutaneous irritants.
EXERCISE XV.— (OPTIONAL) SCARLET RED
Inject an oily solution under the skin of a rabbit. This causes epithelial proliferation
— although not cancer (B. Fischer, 1906).
CHAP. XXXVIII FATE OF DRUGS; IDIOSYNCRASY; EMETICS 211
EXERCISE XVI.— (OPTIONAL) EXPERIMENTAL PLEURISY
Pleurisy with fibrinous exudate may be produced in dogs by injection of i c.c. of oil of
turpentine into the pleural cavity (Opie, 1907, Jour. Exp. Med., 9, 391; 1908, ibid., 10, 423).
A leukocytic exudate is obtained in rabbits by the intrapleural injection of 10 c.c. of 5 per
cent, aleuronat suspension in 3 per cent, starch paste. The animal may be killed and
examined after twenty-four hours.
TECHNICAL REFERENCES
Permeability of Vessels. — Estimation by passage of iodid or ferrocyanid into peri-
toneum, Luithlen, 1913, Med. Klin., No. 42, p. 4.
Differentiation of Exudatcs and Transudates. — Acetic Acid Test, Barberio, 1914, Zentr.
Bioch. Bioph., 17, 450.
CHAPTER XXXVIII
FATE OF DRUGS; IDIOSYNCRASY; EMETICS. (A) ABSORPTION;
(B) EXCRETION; (C) DISTRIBUTION AND INTERACTION OF
DRUGS; (D) IDIOSYNCRASY; ATROPIN THYROID TEST; (E)
EMETICS; (F) ANTEMETICS.
(A) THE ABSORPTION OF DRUGS
Introduction. — Most drugs must be absorbed before they can produce
any action. This holds particularly for drugs which act systemically •, i. e.,
on the body cells (in contradistinction to the locally acting drugs, the effects
of which are confined to the place where they are applied, or to reflexes
originating from this point). The subject of absorption has therefore a
great practical importance. Absorption may occur from most of the sur-
faces of the body, but with very different facility. The inlact skin of
mammals is almost impermeable to watery solutions, but absorbs oils and
volatile substance. The skin of frogs, however, absorbs watery solutions
readily, being rather analogous to mucous membranes. In mammals the
most usual channels of absorption are the alimentary canal, the subcutaneous
and muscular tissue, and the lungs. The rapidity of absorption varies with
the nature of the drug and the place of administration. It is generally
proportional to the volatility and solubility of the drug. Volatile sub-
res are absorbed most rapidly from the lungs; watery solutions from
intramuscular and subcutaneaus injections; resins and oils from the intestinal
trad. The absorbability from the different portions of the alimentary
canal varies for different animals and drugs. It is generally most effect i\ e
from the -mail intestine; less so from the stomach and rectum. The un-
injured urinary bladder i- practically impermeable, while the inucosa of the
urethra i- a u<x>d absorbing surface. Most mucosae absorb readily.
Absorption is retarded 1>\ tin presence of f a ts or colloids, gums, proteins.
actives."
The doses in the comparative experiments must be calculated and
>ured very accurately. The injection >yringe must be wa>hcd with a
little water, which is then also injected.
TECHNICAL NOTES
Stomach-tube.— This consists of a stout, soft gum catheter (No. 10,
'.i>h scale, for dogs), attached to the injection bulb shown in Fig. 43.
212
A LABORATORY GUIDE IN PHARMACOLOGY
The mouth of the animal is held open with a perforated gag, the head
of the animal is bent forward, and the moistened catheter is passed well
back, when no difficulty will be found in making it enter the esophagus.
Care must be taken not to push it into the trachea, and it is well to note
that the animal does not breathe through the catheter. The accident may
also be discovered by the fact that the catheter
cannot be pushed as far and that the fluid flows
in with much greater difficulty. After making
sure that the tube has entered the stomach the
solution is poured into the bulb. If it does not
flow readily, it can be quickened by blowing.
Perforated Gag. — This is shown in Fig. 44. It
is made of hard wood of various sizes. The up-
rights are stiff wire rods, to prevent the animal
from turning its head. A copper wire may be
attached to one rod, brought behind the animal's
ears, and wound around the other rod, thus keep-
ing the gag in place.
The administration per rectum is done with the
same form of apparatus as is used in the stomach.
The catheter should be introduced as high as
possible. The anus is then closed with bulldog
forceps.
Hypodermic injections are generally made under
the loose skin of the flank, the animal being held
securely. The volume of fluid should be kept
below i c.c. for guinea-pigs, and below 5 c.c. for dogs. If it is necessary
to inject larger quantities, they should be given in fractions, distributed
over several parts of the body. The injection of irritant substances should
be avoided.
Fig. 43. — Injection bulb
(made of a capacity of 100
Fig. 44.— Perforated gag.
With dogs and cats the injection is usually made in the back or flank;
with rabbits and guinea-pigs, in the abdomen; with rats and mice, at the
root of the tail.
An ordinary (i c.c.) hypodermic syringe and strong "antitoxin" needle
answers for the smaller quantities; a 5-c.c. antitoxin syringe with an "as-
pirator" needle is used for dogs.
CHAP. XXXVIII FATE OF DRUGS; IDIOSYNCRASY; EMETICS
213
Intramuscular injections are generally made into the gluteal muscles.
Intrapcritoncal and intrnpleural injections are made by thrusting in the
small needle perpendicular to the surface of the body. In making an intra-
peritoneal injection the skin and muscles are pinched in the median line
below the umbilicus.
For intravenous injections a cannula is tied
into a vein, pointing toward the heart, and this
is connected with a buret containing the solu-
tion. The rubber connection should be short
to avoid dead space. It is closed by a Mohr
clamp. If the injection is to be made slowly,
a screw-clamp must be placed on the rubber
tube. The greatest care must be used to
avoid the entrance of air bubbles into the
vein. Before connecting, the rubber tube
should be completely filled with the solution,
and the cannula should also be filled (with a
pipet). If the volume of the injected fluid is
small, it may be introduced at air temperature;
if it exceeds 10 c.c., it should be brought to
body temperature. (Greene advises to sur-
round the buret with the jacket of a Liebig
condenser, through which water of the desired
temperature is circulated.)
The arrangement shown in Fig. 45 may be
used when large quantities of warm fluid are
to be infused in long experiments.
If a number of small injections of different
drugs are to be made in quick succession, it
may be more convenient to clamp the rubber
tube \ inch above the cannula, and to make
the injection with a hypodermic syringe, thrust-
ing the needle obliquely through the rubber
into the cannula.
The injections may be made either into the
femoral or jugular vein. The former is pre-
ferred, as the jugular injection introduces com-
plications by bringing the drug directly into
the heart in too concentrated a form. It may be
necessary in small animals in which it is difficult
to introduce a cannula into the femoral vein.
In umincsthct'rctl rabbits intravenous injections may be made by thrift ing
the needle of the hypodermic syringe into one of the ear vein>. \\hu h ha>
been previously rubbed with xylol and distended by pressure.
Injections into arteries require some pressure. This may be obtained by < onm •« tinn
the t<.p <.t tin- syrinirr with a pressure bottle; or, more convrmmily. with tin- lompressed
nk.
1 quantity l»c injected with an «>r«lin. into the central
nul r.i! artery. Th< H than with ii <• m<l more
than with hy|xi<lcrmi< . a<lminUt ration (Mayor, i.jos
Technical References. — Stomach-tube. — Alxlrrh.iMrn. J, i .\v, 5, uo; guinea-pig, ibid.,
3. *
Abderhalden, 3, 1286; Subcorncal, ibid., 1285.
/w/nr»rw. :. I >:. I'ittcnger, 125.
Fig. 4*— Arrangement for pro-
longed infusion of warm fluids.
214 A LABORATORY GUIDE IN PHARMACOLOGY
Injection into Rabbit's Ear. — Ibid., 3, 1186; 5, 23.
Infusion Under Constant Velocity.— W. Straub, 1911, Muench. Med. Woch., No. 28;
Sansum, Wilder, and Woodyatt, 1916, Proc. Amer. Soc. Biol. Chem., 3, 19.
Inoculation of Precise Amounts. — Rosenau, 1904, Hyg. Lab. Bui. No. 19.
Intestinal Absorption. — Robert, Intox., i, 252.
Syringes. — Abderhalden, 3, 1278; Pittenger, 121.
EXERCISE I.— (DEMONSTRATION) RAPIDITY OF ABSORPTION BY VARI-
OUS CHANNELS: EPINEPHRIN AND STRYCHNIN
(REPORTER V, D)
The evanescent action of epinephrin makes it particularly suitable for
illustrating this subject. Employing uniform doses, the height of the blood-
pressure rise varies directly, the duration of the rise inversely, to the rapidity
of absorption.
Experiment i. Epinephrin. — Arrange an anesthetized dog for blood-pressure tracing;
divide both vagi; connect central end of femoral artery and femoral vein for injection.
While taking slow tracings, inject uniform doses of epinephrin, viz., 0.05 mg. per kg. (0.05
c.c. or one drop of i : 1000 per kg.), as follows, always waiting with the next injection until
the blood-pressure has returned to normal:
(a) Intravenously.
(b) Under mucosa of nasal septum.
(c) Into central end of femoral artery.
(d) Intramuscular.
(e) Into peritoneum.
(/) Into vein.
(g) Into pleura.
(h) Under skin.
(i) Into stomach.
(j) Into vein.
Experiment 2. Strychnin. — Tie the vesico-urethral orifice; inject into bladder a twice
fatal dose of strychnin (1.5 mg. X kg.); in fifteen minutes repeat into ligated stomach; in
ten minutes repeat into intestines.
QUESTIONS
(a) Describe the effects of epinephrin on absorption.
(b) State the order of its absorbability by the various channels.
(c) Are all mucous membranes suited to the absorption of strychnin?
(d) Why was it necessary to tie off the bladder?
EXERCISE II.— (DEMONSTRATION) RAPID ABSORPTION FROM ORAL
ADMINISTRATION: NICOTIN AND HYDROCYANIC ACID
(REPORTER V, D)
While most drugs are absorbed relatively slowly when given by mouth,
the absolute rapidity varies greatly. With nicotin and cyanid the absorp-
tion is almost instantaneous.
Experiment i. Nicotin. — Place i drop of nicotin on the gums of a cat
(or 2 drops for a dog). Note heart-rate and time of evidence of the fol-
lowing symptoms: Excitement; salivation; retching; hyperpnea; prostra-
tion; convulsions; erection of hairs; arrest of respiration; arrest of heart.
Experiment 2. Hydrocyanic Acid. — Inject 2 per cent, of prussic acid
into mouth of rabbit1 or cat (i c.c.) or dog (5 c.c.). Observe as for nicotin
in Experiment i. The effects are very similar, but the heart-rate is not
quickened and the mucous membranes may not be cyanotic.
i Use rabbit of Exercise VIII.
CHAP. XXXVIII FATfc OF DRUGS; IDIOSYNCRASY; EMETICS 21$
Jnarirr
QUESTIONS University
(a) Describe the symptoms of poisoning by (i) nicotin, (2) hydrocyanic
acid.
(b) How soon do they appear with (i) and (2)?
(c) How rapidly are they fatal with (i) and (2)?
(d) Which stops first, heart or respiration, with (i) and (2)?
(e) What difference is there between the color of the mucosae with the
two poisons?
;) How is this explained?
) What symptomatic treatment would you suggest for these poisons?
(h) Discuss its probable efficiency.
EXERCISE m.— (DEMONSTRATION) RAPID ABSORPTION OF GASES BY
LUNGS
(REPORTER V, D)
The extensive surface of the alveolar capillaries insures rapid absorption
of vapors, provided that they reach the alveoli, and that the epithelium is
not impermeable to them.
Experiment i. Carbon-monoxid Poisoning. — Place a guinea-pig, rat, or
other small mammal under a bell-jar and pass coal-gas into the jar: the
animal shows almost immediately signs of asphyxia; uneasiness; inco-
ordinated convulsions (medullary type) ; coma; dilated pupils. The mucous
membranes, however, are not cyanotic. Remove from bell-jar and start
artificial respiration : prompt recovery.
QUESTIONS
(a) Describe the symptoms of coal-gas poisoning.
(b) Why are the effects so rapid?
(c) Which constituent of the gas is mainly responsible?
(d) How does it act?
(e) How do the effects differ from those of ordinary asphyxia by oxygen
deprivation?
SWhat should be the treatment for poisoning by the asphyxiant gas?
What would be the advantage, if any, of administration of oxygen?
EXERCISE IV.— (GROUP I) ABSORPTION OF STRYCHNIN FROM ORAL
AND HYPODERMIC ADMINISTRATION
D
Give to a rabbit (A) i.o mg. JUT kg. of strychnin sulphate d.o c.c. per
i ,'„ prr cent.) hypodermic ally, and to another rabbit (B> tin sime
amount by the stomach-tube. Tin- tir>t rabbit shows the typical itiyn linin
convulsions; the second shows very little effect. Draw a sketch of t In-
typical tetanic condition.
QUESTIONS
(a) Describe the Mrydmin symptom-, their onset and duration.
(b) Would a fatty or resinous substance also be absorbed more readily
I'mm hvpo.l, rmie than from oral administration? Why?
(c) Would it be probable that a definite ratio between hypodermic and
oral dosage could be established that would U- valid for all drugs?
2l6 A LABORATORY GUIDE IN PHARMACOLOGY
EXERCISE V.— (GROUP H) ABSORPTION OF CHLORAL FROM ORAL AND
RECTAL ADMINISTRATION
(REPORTER V, D)
Use two weighed cats. Administer to (A) chloral, 0.25 gm. per kg.
(2.5 c.c. of 10 per cent, per kg.), by stomach-tube; to (B) the same dose by
rectum. Note onset of symptoms — drowsiness, ataxia, anesthesia, etc.
Compare temperature and respiration at the end of the experiment.
QUESTIONS
(a) Describe the symptoms of chloral poisoning.
(b) Which is the more efficient channel of absorption?
(c) Would there probably be a constant ratio for all drugs?
EXERCISE VI.— (GROUP III) COLLOID ON ABSORPTION (STRYCHNIN
AND STARCH)
(REPORTER V, D)
Use two weighed cats. Administer by stomach-tube to cat (A) strych-
nin, i mg. per kg. (i c.c. of i : 1000 per kg.), diluted with 10 parts of
water. Administer to cat (B) the same dose, but diluted with 10 parts
of 25 per cent, acacia. Compare the onset and severity of the convulsive
symptoms.
QUESTIONS
(a) Describe the strychnin symptoms.
(b) What is the influence of the colloid on absorption?
(c) Would a given quantity of strychnin be as active when given in the
form of tincture of mix vomica as if it were given pure?
(d) Would acacia or starch paste be of any value in strychnin or similar
poisoning?
(e) What would be its limitations?
(B) THE EXCRETION OF DRUGS
Introduction. — Drugs may be excreted by various channels: gases
and volatile drugs are excreted mainly by the lungs ; metals by the intestinal
cells; most substances, however, especially salts and alkaloids, are excreted
in greatest quantity by the urine. The saliva, bile, skin, and milk may also
aid in excretion; but generally these play a very subordinate role.
A knowledge of the excretion of drugs has considerable practical im-
portance; it teaches how frequently the drug must be administered to main-
tain a continuous action; it also indicates how to hasten the elimination
of poisons.
The elimination of drugs by the urine and saliva was studied in Exercise
XV. This should be reviewed.
EXERCISE VH.— (DEMONSTRATION) PULMONARY EXCRETION (H2S)
(REPORTER I, A)
Hold a paper saturated with lead acetate before the nostrils of a rabbit
or cat; note that the paper is not blackened; now pass some H2S into the
rectum: the paper becomes blackened (the H2S being absorbed from the
rectum and excreted by the lungs). If the dose of H2S has been excessive,
the rabbit may show paralytic and convulsive effects. (The experiment is
CHAP. XXXVIII FATE OF DRUGS; IDIOSYNCRASY; EMETICS 217
not quite conclusive, for the gas might have reached the paper through
the esophagus.)
Not all gases, however, are capable of excretion by the lungs; for in-
stance, ammonia is not excreted by an uninjured lung.
(C) DISTRIBUTION AND INTERACTION OF DRUGS
Introduction. — The distribution of drugs within the body -follows special
laws, differing for individual drugs. It is also affected by disease, as illus-
trated by the use of fluorescein in ophthalmic diagnosis. Several drugs may
react within, as well as outside of the body, as shown by the calomel-iodid
experiment.
A considerable number of drugs undergo chemic changes during their
sojourn in the body, being oxidized, reduced, hydrolysed, combined, etc.
In some cases the substance is absolutely destroyed. Alcohol, for instance,
is almost completely oxidized to carbonic acid and water. In other cases
the changes are not so profound. The benzol ring tends to remain intact,
but the transformation of acetanilid into paramidophenol illustrates the
changes which occur in the side-chains. Benzol derivatives are further
excreted as paired compounds, with sulphuric and glycuronic acid.
EXERCISE Vin.— (DEMONSTRATION) IODID, MORPfflN, CALOMEL,
FLUORESCEIN
(REPORTER I, A)
A rabbit has received, two hours before the demonstration, 50 c.c. of
i per cent, sodium iodid by stomach-tube and 20 mg. of morphin per kg.
(0.5 c.c. per kg. of 4 per cent.) hypodermically. An hour before the demon-
stration some calomel was dusted on the conjunctiva of one eye. Calomel
is also applied to an eye of a normal rabbit.
The animal will present the symptoms of morphin poisoning. The
iodid as such produces no visible effects; but the calomel on the eye of the
iodid rabbit shows intense congestion and edema, and probably the yellow
color of mercuric iodid. The calomel has produced no effect on the eye
of the normal rabbit.
The eyes are washed. Into those of the iodid rabbit is dropped some
fluorescein solution (fluorescein, 2; sod. bicarb., 3; water, 100). This
is left for two minutes, and the eyes are then rinsed with water. Any
lesions of the cornea will be stained yellow, while normal tissue remains
unstained.
(The animal shall be killed before it recovers from the morphin.)
QUESTIONS
(a) Describe the effects of morphin.
(b) Why i> it dangerous to use calomel with iodid?
(c) Describe the fluorescein test.
(d) Why does fluorescein stain the ulcerated corneas and not the normal?
EXERCISE IX.— (OPTIONAL) DISTRIBUTION AND EXCRETION OF HEXA-
METHYLENAMIN
•c a dog with nutrpliin ;m<l rtlu-r. I'l.ur ..mmil.i into «>nc un tcr. Ad-
minUtrr hex. in ,n, 0.5 urn. |>rr kv... <li«.M,l\,-«l in water, l.\ itOOUM h b
•lie urine flow ini DOtC time \\hcn turbid-
ity first appears COITC*IM aiding to the beginning hrx.um tin It n.unin.
2l8 A LABORATORY GUIDE IN PHARMACOLOGY
At fifteen-minute intervals test for presence of hexamethylenamin in urine, blood, and
saliva (see page 70). Note the relative intensity of the reaction.
After two to four hours kill the animal and collect the bladder urine, the bile, the
pleural, peritoneal, cerebrospinal and synovial fluid, and the aqueous humor. Apply
tests for hexamethylenamin and for free formaldehyd. Formulate conclusions.
QUESTIONS
(a) In what situations is the hexamethylenamin found?
(6) How soon does it appear?
(c) What is its relative concentration?
(d) Where is formaldehyd formed from it?
TECHNICAL NOTES AND REFERENCES ON CEREBROSPINAL FLUID
Continuous Collection of Cerebrospinal Fluid (Dixon and Halliburton, 1913, Jour.
Physiol., 47, 218): "The skin at the back of the neck and about an inch from the occipital
process is severed for about i cm.; the sub-cerebellar cisterna is then punctured by means
of a trocar and wide cannula, shaped in the usual way, but with a blunt point. The easiest
way of performing this is to flex the animal's head as far as possible and insert the trocar
with its point directed to a spot mid way between the eyes: it should then pierce the occipito-
atlantoid ligament and enter the foramen magnum. The forward movement of the trocar
should cease as soon as active resistance to its movement ceases. On the withdrawal of
the trocar the clear cerebrospinal fluid gushes out entirely free from blood.
"There is no necessity to tie the cannula in any way, since it is firmly fixed in the com-
pact tissues in the back of the neck. A glass tube is now connected to the cannula by a
short rubber connection and the cerebrospinal fluid is allowed to drip into a glass capsule.
The fall of each drop is signalled electrically on the base line of the arterial pressure which
is simultaneously taken."
Obtaining Cerebrospinal Fluid. — Tigerstedt, 3.4, 133; Weed and Gushing, 1915, Amer.
Jour. Physiol., 36, 77; Chemic Examination, Abderhalden, 5, 215.
Lumbar Puncture. — Tigerstedt, 3.4, 8.
Artificial Hydrocephalus. — Frazier and Peet, 1914, Amer. Jour. Physiol., 35, 268.
EXERCISE X, A.— (OPTIONAL) ABSORPTION AND EXCRETION ON EFFECT
(POTASSIUM CHLORID)
Anesthetize a dog with morphin and ether. Arrange for blood-pressure tracing. Ligate
pylorus. Inject by stomach-tube KC1, 2 gm. per kg., diluted with water. Note that the
blood-pressure does not change materially within an hour. Now ligate the renal vessels
and repeat the KC1: the blood-pressure falls gradually.
QUESTIONS
(a) Why is the potassium ineffective by stomach?
(b) How can it be made effective? Why?
EXERCISE X, B.— (OPTIONAL) ABSORPTION INTO BLOOD AND LYMPH
Anesthetize an animal and place cannula into the ureters and thoracic duct. Inject
methylene-blue solution into the peritoneal or pleural cavity: the color appears in the
urine before the lymph (Starling and Tubby).
QUESTION
Does the absorption of the dye from serous cavities occur by the blood or lymph?
(D) IDIOSYNCRASY; ATROPIN; THYROID TEST
Introduction. — Idiosyncrasy is the term applied to an abnormal reaction
to a drug. The abnormality is generally quantitative only; but it may
appear qualitative by bringing into prominence some action of the drug
CHAP. XXXVIII FATE OF DRUGS; IDIOSYNCRASY; EMETICS 2IQ
which is ordinarily so small as to escape observation. Most instances of
idiosyncrasy may therefore be brought under the headings of exaggerated
susceptibility or tolerance. These may be congenital or acquired. Some
are readily explained by anatomic or physiologic peculiarities. Others
are due to differences in the absorption, excretion, or destruction of the
poison. Many phenomena of idiosyncrasy have not yet been satisfactorily
explained. The continued administration of a drug often alters the sus-
ceptibility of the patient to its action; this may be diminished (habituation)
or increased (cumulative action). Congenital idiosyncrasy may be individual
or racial. The student will probably encounter some examples of individual
idiosyncrasy in the course of his future work. The following experiments
refer mainly to racial idiosyncrasy.
EXERCISE XI.— (GROUP IV) ATROPIN ON DOG AND RABBIT
(REPORTER I, A)
On a dog (A) and rabbit (B) observe the normal pulse, pupils, and respi-
ration. Confirm also that the rabbit reacts to inhalation of ammonia by
temporary arrest of the heart (trigeminal-vagus reflex).
Inject each animal, hypodermically, with Atropin, 5 mg. per kg. (0.5 c.c.
of i per cent, per kg.). Repeat the observations from time to time. The
effects are very much greater in the dog than in the rabbit (the heart
rate is quickened by paralysis of the vagus endings; the respiration is
first increased, then diminished. The general symptoms are first ex-
citant, later depressant; the pupils are dilated through paralysis of the
oculomotor endings). Let the rabbit inhale a little ammonia while feeling
the heart-beat: the heart is not stopped, as it would be in normal animals.
It will have been noted that the pulse-rate is greatly quickened in the dog,
but scarcely, if at all, in the rabbit. This is because in the dog the heart
is normally kept slow by the tonic activity of the vagus center. This is
cut out by atropin. These tonic impulses are very weak or absent in the
rabbit, so that their abolition does not alter the heart-rate. The ammonia
experiment shows that the atropin has paralyzed the vagus in the rabbit as
WL-11 as in the dog.
The general resistance of rabbits is due, at least partly, to the more
rapid destruction of atropin in their tissues.
QUESTIONS
(a) Describe the effects of atropin on general behavior; pulse; pupils;
respiration.
(b) Why does the effect on the pulse clitTrr in dogs and rabbit-?
(c) How can it be shown that the vagus is paralyzed in rabbit-?
EXERCISE XII.— (OPTIONAL) DIGITALIS ON TOAD AND FROG HEART
Apply some (10 prr «-nt.) infu-ion of ,/;\-//<;//.v in 0.75 NuCl solution to the exposed
• if a piihcd toad and frnu. and noti.c that tin- rlTo t on tin- fn>n i- mu
(Observe that tl m. rr.isnl. prrM;ilti« \va\rs and arhvthniia
become apparent, and the heart may be arrested in systolic standstill as a small \\lntc
d secretes a poison \\ith an a. ti..n .•• to digitalis. The
ncc of this animal is therefore somewhat analogous to habituation.
220 A LABORATORY GUIDE IN PHARMACOLOGY
EXERCISE Xffl, A.— (OPTIONAL) ACETONITRIL TEST FOR THYROID
(HUNT'S METHOD)
The feeding of thyroid to mice greatly increased their resistance to acetonitril, pre-
sumably by diminishing the liberation of HCN. This serves as a qualitative and even
quantitative test for thyroid substance.
White mice are fed for some weeks on a uniform diet (oats and water). The M. F. D.
of acetonitril, hypodermically injected, is ascertained (beginning with 0.25 mg. per gm.,
fresh solution). This serves as a control. The thyroid preparation (say i mg. per day
per mouse) is made into pills with cracker-dust and syrup, and these are added, for about
ten days, to the diet of some mice of the same lot kept under the same conditions. At
the end of this time the M. F. D. of acetonitril is determined on these mice (starting with
3, 6, 9, 12 times the normal dose).
EXERCISE XHI, B.— (OPTIONAL) TADPOLE TEST
Feeding of thyroid to tadpoles hastens their development, but checks growth (Guder-
natsch, 1912, Arch. Entwickl., 35, 457). Marine and Feiss, 1915 (Jour. Pharmacol., 7,
57 2), perform the test by feeding five tadpoles with 50 mg. of powdered thyroid every other
day; on the alternating days the animals are fed with fresh sheep liver for two hours.
TECHNICAL REFERENCES
Acetonitril Test. — Hunt, 1909, Hyg. Lab. Bui. No. 47; Fuehner, 153.
Mice. — Keeping and breeding: Abderhalden, 3, 1268; Anesthesia, ibid., 3, 1281;
Injection, Fuehner, 148.
Thyroidectomy. — Abderhalden, 6, 560.
Thyroid Experiments. — Robert, Intox., i, 267.
Cretinism. — In young rats, by complete thyroidectomy, Basinger, 1916, Arch. Int.
Med., 17, 260.
(E) EMETICS
Introduction. — These illustrate phases of racial idosyncrasy, but the
subject has also a direct practical importance.
Emetics are divided into two classes : Those which stimulate the vomit-
ing center in the medulla directly (central emetics) and those which stimulate
it reflexly (local emetics). The central emetics act at least equally well
when they are injected hypodermically. Apomorphin is the principal
example. Local emetics act by irritating the sensory endings in the pharynx
or stomach. They are effective only if they are administered or excreted
by this channel. All irritants belong to this class; but only those are
practically useful which have only a slight toxicity, or which act so 'promptly
that they are expelled before absorption can occur.
If a drug produces vomiting when injected into the circulation, and
not when it is given by mouth, its action is surely central; and vice versa.
If it causes emesis in either case, the relative quantity and the time re-
quired are taken into consideration: if it is more efficient by the circulation,
its action is, at least mainly, central; and vice versa. The absolute dis-
tinction is made by ligating all the vessels of the stomach, exclusive of the
nerves: a centrally acting emetic will now be effective only when injected
into the circulation, a local emetic only when placed in the stomach.
Emesis, the act of vomiting, is preceded by nausea, and followed by
depression. The relative duration of these stages is of great practical
importance.
Observations to be Made. — The onset and duration and symptoms of
nausea; onset and frequency of emesis; pulse and respiration of normal
animal in nausea, just before, during, just after, and some time after
vomiting. Note how soon the animal will drink water and eat meat again.
Report the results. The animals should have been recently fed.
CHAP. XXXVIII FATE OF DRUGS; IDIOSYNCRASY; EMETICS 221
EXERCISE XIV.— (GROUP V) APOMORPfflN
(REPORTER II, A)
Inject Apomorphin hypodermically as follows, and observe effects:
Dog A — i mg. (o.i c.c. of i : 100) per kg.
Cat B — 5 mg.. (0.5 c.c. of i : 100) per kg.
Cat C — 50 mg. (5 c.c. of i : 100) per k
Rabbit D — 10 mg. (i c.c. of i : 100) per kg.
(Optional) Apomorphin as Hypnotic. — This effect may be produced, rather uncertainly,
by small doses (0.04 mg. per kg. for cats or dogs, hypodermically).
QUESTIONS
(a) Describe the phenomena of apomorphin-vomiting as witnessed in
Dog A and Cat C.
(b) Describe the phenomena of apomorphin-nausea as witnessed in
CatB.
(c) Describe the phenomena of apomorphin excitement as witnessed in
Rabbit D.
(d) Why does the rabbit fail to vomit?
(e) Why is the cat less susceptible to apomorphin? Is it generally
resistant to emetics?
(/) Is the action of apomorphin central or local? How could this be
proved?
EXERCISE XV.— (OPTIONAL) LOCATION OF APOMORPfflN ACTION
(See Eggleston and Hatcher, 1912, Jour. Pharmacol., 3, 551.)
EXERCISE XVI.— (GROUP I, A, H, A, m, A) LOCALLY ACTING EMETICS
(REPORTER II, A)
Administer the following solutions to cats (or dogs, with double dose)
by stomach-tube:
(Group I, A) Copper Sulphate, 25 c.c. of i per cent.
(Group II, A) Zinc Sulphate, 25 c.c. of i per cent.
(Group III, A) Tartar Emetic, 10 c.c. of J per cent.
(Optional).— Ipecac (i c.c. of fiuidrxtnu t '. Mu>tanl (teaspoonful) in warm
Ammonium ( 'arbonate (10 c.c. of 5 per cent, solution). Senega (2 c.c. of lluidcxtract).
QUESTIONS
Describe the phenomena of nausea and vomiting; onset and duration;
amount of depression.
F. ANTEMETICS
Kme-i- may IK- treated cither liy <leprc»inu the vomiting n-nter OF
(with the locally acting emetics) by protecting the Momach a^ainM local
irritation.
EXERCISE XVII.— (GROUP I, B AND II, B) PARALYSIS OF VOMITING
CENTER
II. A)
Experiment i. (Group I, B) Morphin and Apomorphin.-- Inject into
dog 10 mg. per kg. (J c.c. p« t > of morphin, subcutaneous! y.
222 A LABORATORY GUIDE IN PHARMACOLOGY
This will cause vomiting, probably by an action similar to apomorphin
(which is a derivative of morphin). After half an hour inject apomorphin
i mg. (o.i c.c. of i per cent.) per kg. hypodermically. This will be ineffective,
as the morphin stimulation of the vomiting center is followed by a profound
depression. All other emetics will be similarly ineffective. This is utilized
in experimental technic, when it is essential to have an irritant drug retained
in the stomach.
Experiment 2. (Group II, B) Morphin and Zinc Sulphate. — Proceed as
in Experiment i (dog or cat), but use 50 c.c. of i per cent, zinc sulphate
(25 c.c. for cat) by stomach- tube in place of the apomorphin.
QUESTIONS
(a) How does morphin affect emesis?
(b) Does it act against both central and local emetics?
(c) Would it be available as a therapeutic measure?
EXERCISE XVHI.— (GROUP HI, B) BISMUTH AND ZINC SULPHATE
(REPORTER II, A)
Administer to a cat, by stomach-tube, i gm. of bismuth subcarbonate,
suspended in 50 c.c. of mucilage of acacia. In ten minutes follow this by
25 c.c. of i per cent, zinc sulphate, also by stomach-tube: vomiting will be
delayed or prevented.
QUESTIONS
(a) Would bismuth be effective against both classes of emetics (central
and local)?
(b) Could it be used clinically? Against which conditions?
TECHNICAL REFERENCES ON DIGESTIVE TRACT
Operations. — London in Abderhalden, 3, 76.
Digestion Experiments on Animals. — Zunz, ibid., 3, 122.
Collection of Digestive Secretions. — Ibid., 3, 189.
Products, Collection, and Analysis. — Ibid., 6, 458.
Examination of Stomach Contents. — Abderhalden, 8, 44.
Indicators of Gastric Acidity. — Fowler, Bergheim, and Hawk, 1915, Soc. Exp. Biol.
Med., 13, 58.
Aseptic Technic. — Tigerstedt, i.i, 55; 3.4, 16; Hoskins and Wheelan, 1914, Amer.
Jour. Physiol., 34, 81. lodin as skin disinfectant, Mayers, 1911, Soc. Exp. Biol. Med.,
8, 53-
Digestive Fistula. — Permanent: Ibid., 6, 564; Thiry-Vella, ibid., 6, 466.
Eck's Fistula. — Abderhalden, 6, 528; Peet, Ann. Surg., Nov., 1914.
Pancreas Extirpation. — Asher, 1914, Zs. biol. Tech., 3. No. 6; Hedon, 1911, Arch.
Internal. Physiol., 10, 350.
Pancreatic Juice. — Abderhalden, 6, 488.
Pancreas Experiments. — Robert, Intox., i, 265.
Bile Secretion. — Ibid., i, 262; Exclusion from Intestine, Pearce and Eisenbrey, Amer.
Jour. Physiol., 32, 417.
Evisceration of Animals. — Barcroft and Brodie, 1904, Jour. Physiol., 32, 19.
Splenectomy. — Abderhalden, 6, 561.
CHAP, xxxix METABOLISM; DEPRESSANTS; IRRITANTS 223
CHAPTER XXXIX
METABOLISM; DEPRESSANTS; IRRITANTS. (A) TEMPERATURE;
(B) GLYCOSURIA; (C) METABOLISM; (D) CENTRAL DEPRESS-
ANTS AND TREATMENT OF DEPRESSANT POISONING; (E)
GASTRO-ENTERITIS; (F) NEPHRITIS; (G) REFLEX EFFECTS
OF IRRITANTS. ARSENIC ON BLOOD-PRESSURE
(A) EFFECTS ON TEMPERATURE
Introduction. — The temperature of an animal is determined by the
relation of heat dissipation and heat production. The heat-regulating
mechanism of warm-blooded animals is able to keep the temperature of
the body constant, notwithstanding all ordinary variations of external
and internal conditions. The temperature can therefore be altered only
by very violent changes or, more commonly, by disturbing the regulating
mechanism. Several centers are concerned in the latter. Successive
stimulation or section of the paths is necessary to distinguish which of
these is concerned in a given phenomenon. These experiments are rather
complicated.
By the use of the calorimeter and by the study of metabolism it is easy
to determine whether the change of temperature is due to altered heat
production or heat loss. The plethysmograph will show whether changes in
heat loss are due to an action on cutaneous vessels. The evaporation
of sweat may be excluded by atropin, which paralyzes the sweat-glands.
The drugs which increase temperature act generally on heat production
by increasing muscular movement. Cocain acts on the centers of the
caudate nuclei. The hypodermic injection of irritants, even of water,
and especially of albumose, produces some hyperpyrexia in rabbits. Bac-
terial toxins are the most efficient pyretics.
The drugs which lower temperature may do so by producing a general
depression of the central nervous system — a shock or collapse action.
Alcohol, chloral, morphin, etc., belong to this class. These lower the
temperature even in previously healthy individuals.
The typical antipyretics, on the other hand, lower the temperature
only when it is abnormally hi^h, /. r., in fever; and then only to normal.
The coal-tar antipyretics (acetanilid, antipyrin, etc.) act centrally, and
increase the heat dissipation by dilating the cutaneous capillaries. Quinin
diminishes the heat production by a direct action on the muscular metab-
olism.
Observations Required. — Rectal temperature (every hall -hour). The
observations should be made before giving the drugs, and the animal
should be kept and observed under perfectly uniform conditions, to exclude
accidental variations. (The effects are usually >een in two to three hours.)
The thermometer must be oiled and inserted aluay- to the same depth
(2 or s inches). With small animals the bulb should be warmed in the hand.
I\alil»it> have a more responsive temperature than cats, but these may be
substituted, using the same doses per kilogram. Plot curves of the tem-
perat
TECHNICAL REFERENCES
Temperature of Rabbits. / , Arch. exp. Path., 72, 97; Normal
Variation! S 3.
General Discussion. — Robert, Intox., i, 200, 280.
224 A LABORATORY GUIDE IN PHARMACOLOGY
Heat Puncture. — Tigerstedt, 3.4, 86; Aaronsohn and Sachs, 1885, Arch. ges. Physiol.,
37, 232; Gottlieb, Arch. exp. Path., 26, 422; Jacobj, Exp. Ther., 151.
Beta-Tetrahydronaphthylamin. — Jonescu, Arch. exp. Path., 60; Elliott, 1914, Quart.
Jour. Med., 7, 120, claims that rabbits show gastric ulcers a few hours after hypodermic
injection.
Calorimetry. — Abderhalden, 3/1158; 7, 658; Tigerstedt, 1.3, 150; Lusk, 1915, Arch.
Int. Med., 15, 793; Small Animals, Langworthy and Milner, 1916, Jour. Agr. Res., 6,
703-
Clinical, Gephart and DuBois, 1915, Arch. Int. Med., 15, 829; Human, Langworthy
and Miller, 1915, Jour. Agr. Res., 5, 299.
Surface Area Measurement. — Man, DuBois and DuBois, 1915, Arch. Int. Med., 15,
868; F. G. Benedict, 1916, Amer. Jour, Physiol., 41, 275.
Heating of Carotid Blood. — Stewart, 297.
Experimental Infections. — Rheumatic Arthritis, Klotz, 1914, Cleve. Med. Jour., 13,
210; Rothschild and Thalhimer, 1914, Jour. Exp. Med., 19, No. 5; Synovitis, Andrei, 1913,
Zentr. Bioch. Bioph., 16, 341 ; Pneumonia, Lamar and Meltzer, 1910, Soc. Exp. Biol. Med.,
7, 102; Wolfenstein and Meltzer, 1912, Jour. Exp. Med., 16; Kline and Winternitz, 1915,
ibid., 21, 304; Kline and Meltzer, 1915, Soc. Exp. Biol. Med., 13, 29 (unorganized sub-
stances); Sisson and Walker, 1915, Jour. Exp. Med., 22, 747 (Friedlander type); Syphi-
lis, Rabbits, Jacobj, Exp. Ther., 99; Trypanosomes, Abderhalden, 5, 1371.
Scurvy, Experimental. — L. Jackson and Moore, 1916.
EXERCISE I.— (GROUP I, A) CHLORAL (FALL OF TEMPERATURE BY COL-
LAPSE)
(REPORTER III, A)
Administer by stomach-tube to cat chloral, 0.5 gm. (20 c.c. of 2.5 per
cent.) per kg. : there is a fall of temperature, general depression, and partial
or complete coma. The respiration is slower and more shallow. (Depres-
sion of medullary centers.)
EXERCISE II.— (GROUP H, A) MORPHIN (FALL OF TEMPERATURE BY
DIMINUTION OF METABOLISM, AND PERHAPS BY A SPECIFIC EFFECT
ON TEMPERATURE CENTERS)
(REPORTER III, A)
Inject hypodermically into rabbit o.oi gm. per kg. (f c.c. per Kg. of
4 per cent, solution). The effects resemble those of chloral, but are not so
severe. (Test urine for sugar.) A respiratory tracing may be taken if the
animal shows Cheyne-Stokes respiration.
EXERCISE in.— (GROUP III, A) SANTONIN (FALL, THEN RISE)
(REPORTER III, A)
Inject into the stomach of a rabbit 0.5 gm. per kg. of Santoninate of
Sodium (10 c.c. per kg. of 5 per cent.) : there is at first a fall of temperature
due to the increased heat loss. Convulsions set in, and when these are
violent the temperature may rise on account of the increased muscular
activity. When the convulsions give place to paralysis there is a second
more profound fall of temperature. (Santonin illustrates typically the
effect of all convulsant poisons on temperature.)
EXERCISE IV.— (GROUP IV, A) COCAIN (RISE OF TEMPERATURE THROUGH
STIMULATION OF THE CAUDATE NUCLEUS)
(REPORTER III, A)
Inject hypodermically into rabbit, cat, or dog cocain, 25 mg. (0.5 c.c.
of 5 per cent.) per kg. : rise of temperature of i° to 2° C. The animal may
show great excitement and even violent convulsions.
CHAP, xxxrx METABOLISM; DEPRESSANTS; IRRITANTS 225
EXERCISE V.— (GROUP IV, B) BETA-TETRAHYDRONAPHTHYLAMIN
(REPORTER III, A)
Inject hypodermically into rabbit 25 to 50 mg. (^ to i c.c. of 5 per cent.)
per kg. : rise by strong cutaneous vasoconstriction and increased movements.
EXERCISE VI.— ALBUMOSE FEVER AND ANTIPYRETICS
(REPORTER III. \
Use rabbits or, if necessary, cats.
Experiment i. (Group V, A) Albumose (Rise of Temperature).— Inject
hypodermically i gm. per kg. (5 c.c. per kg. of 20 per cent.): rise.
Experiment 2. (Group V, A) Antipyrin (Little Effect on Normal Animals).
— Give o.i gm. per kg. (10 c.c. per kg. of i per cent.) hypodermically.
There is little, if any, effect.
Experiment 3. (Group V, B) Antipyrin in Fever (Regulation of Tem-
perature). — Give albumose, as in Experiment i, and follow this in two to
four hours by antipyrin (as in Experiment 2) . The temperature soon returns
to normal, while that of Experiment i remains high.
QUESTIONS
(a) State which drugs raise, and which lower, temperature.
(b) Are the antipyretics equally efficient in the absence of fever?
EXERCISE VH.— (OPTIONAL) ASSAY OF ANTIPYRETIC EFFICIENCY
(See Kiliani, 1910, Arch. Internal: Pharmacodyn., 20, 333; Fuehner, 157.)
(B) GLYCOSURIA
Introduction. — The presence of sugar in the urine may be due to
several different causes. These are discussed in text-books of physiology.
The presence of reducing substance in the urine, after the adminis-
tration of drugs, is often due to glycuronic acid, which is generally excreted
in paired combination with the drug. These urines reduce Fehling's
solution, but do not give the fermentation test.
The following are examples of drugs that cause the appearance of glycuronic acid:
Copaiba, Chloral. Menthol, Thymol, many volatile oils, Carbon -monoxid, Chloroform,
1 >\ali< Aiid. Benzaldehyd, Morphin.
True glycosuria tin \vhith tin- urine also gives the fermentation test) is caused by:
Phlorhi/in, Kpinephrin. I'ranium, Curare. Cyanids. Atropin. Amyl Nitrite. Chromates
and Bi.hromates, Men ury. Morphin, Cantharidin. extensive >alt injo lions. .
Many of thr>r a. l by producing asphyxia. I'hlorhi/in acts direUh on tin- kidney
TECHNICAL NOTES
Cathctcrization requires considerable- pra.ti.r in do^s and in female rabbits
easy in male rabbit-. A No. 5 bone tipped CUO male catfotH !• used. The in;-
rabbit- may be collet ted by CXprcs .\imi: The animal is ^ra-ped tirmly in the left hand, spas
to pu>h the abdominal organs toward the pelvis, when moderate pressure with tin-
hand. over the bladder, usually a. . omplMu •- the desired result. The urine and fcccs
may also be collected by placing the animals in suitable Metabolism cages.
TECHNICAL REFERENCES
Catheterization of Animals. — Stewart, 690; Abdcrhaldm, 3, 1045; Separation,
Tschermi. ho\\ski. KJOO. Bio, h. Cbl., 8.932.
Glycosuria. Stewart, (xjo; Abdcrhaldcn, 5, 1199.
226 A LABORATORY GUIDE IN PHARMACOLOGY
Sugar Estimation. — Abderhalden, 2, 167 (in blood, ibid., 5, 172); Shaffer, 1914, Jour.
Biol. Chem., 19, 285; Lewis and Benedict, 1915, ibid., 20, 61; Macleod, "Diabetes," 6,
ii, 16, 26; Clinical, Kleiner, 1914, Jour. Amer. Med. Assoc., 62, 1307; Comparison of
Methods, Fitz, 1914, Arch. Int. Med., 14, 133; Morris, 1916, Jour. Lab. Clin. Med., i, 252;
Normal Human Urine (qualitative), Folin, 1915, Jour. Biol. Chem., 22, 327; Determination
of Small Amounts in Urine, V. C. Myers, 1916, Proc. Soc. Exp. Biol. Med.
Glycuronic Acid.— Abderhalden, 2, 101, 139; 3, 949, 9^95 in blood, ibid., 5, 177; paired,
ibid., 6, 258.
Acetonuria, Experimental.— (Phlorizin with fasting), Jacobj, Exp. Ther., 143.
Diabetes Insipidus, Experimental.— S. A. Matthews, 1915, Arch. Int. Med., 15, 451.
EXERCISE Vm.— (GROUP II, A) MORPHIN (ASPHYXIAL CONVERSION
OF GLYCOGEN INTO GLUCOSE) (See EXERCISE II)
EXERCISE IX.— (OPTIONAL) PHLORHIZIN
(Renal action.) Inject hypodermically into a rabbit £ gm. of phlorhizin dissolved in
5 c.c. of warm water. Keep the animal in a cage arranged for the collection of urine. If
none has been passed in an hour, withdraw by a catheter, and demonstrate the presence
of sugar by Fehling's or Trommer's tests.
EXERCISE X.— (OPTIONAL) EPINEPHRIN
Inject subcutaneously into a rabbit i to 2 c.c. of i : 1000 epinephrin; in two hours
collect the urine and test for sugar.
(C) METABOLISM
Introduction. — Drugs may alter metabolism directly by acting on the
tissues or on certain nervous centers; or indirectly by influencing digestion,
absorption, or excretion; or by making the animal quiet or restless, etc.
The experimental investigation of nitrogen or carbon metabolism
entails extensive preparation and surveillance of the animals and time-
consuming analytic methods. The following experiments are, therefore,
optional.
EXERCISE XL— (DEMONSTRATION) ACID INTOXICATION
(REPORTER V, A)
Administer to a rabbit, by stomach-tube, 100 c.c. of i per cent. HC1
per kg.: unsteady motions, slowed heart and respiration, stupor, coma, con-
vulsions, air-hunger, but no cyanosis. Death may occur in twelve to
forty-five minutes. Just before death insert a cannula into the jugular
vein, toward the heart, and inject slowly a 0.5 per cent, solution of sodium
carbonate: recovery.
EXERCISE XII.— (OPTIONAL) EXPERIMENTS ON NITROGEN METABOLISM
Dogs or rabbits may be used. Arrange for the regular collection of urine. The
animals may be reduced to nitrogen equilibrium and then kept on a uniform diet; or they
may be starved until the nitrogen is practically constant. The urine may be examined for
total nitrogen and for urea. The following drugs may be tried:
Quinin: 0.05 per kg.
Antipyrin: 0.2 gm. per kg.
Water: large quantity.
The following drugs are important: Quinin diminishes nitrogen metabolism; the
coal- tar antipyretics also, but only in fever. Morphin diminishes carbon metabolism.
Phosphorus in toxic doses increases nitrogen metabolism, but diminishes urea. Acids
increase ammonia excretion at the expense of urea; alkalies the reverse. Salts and water
increase nitrogen excretion.
CHAP, xxxrx METABOLISM; DEPRESSANTS; IRRITANTS 227
TECHNICAL REFERENCES
Metabolism. — Robert, Intox., i, 208. Experiments on man, Abderhalden, 3, 994.
Utilization of food, ibid., 1002. Protein metabolism, ibid., 1005. Carbohydrates and
fats, ibid., 1009. Nuclein, ibid., ion. Salt, ibid., 1013. Water, ibid., 1014. Energy,
ibid., 1115. Caloric Requirements, ibid., 095. Respiratory, ibid., 1143. Intermediary,
ibid., 5, 1148. Sucklings, ibid., 3, 1016.
Respiration Chamber. — Man, Langworthy and Milner, 1915, Jour. Agr. Res., 5, 299.
Small animals, Rolls and Lpewenhart, 1915, Amer. Jour. Physiol., 39, 67; Benedict, 1915
(COi and oxygen), Jour. Biol. Chem., 20, 301.
Animals, Apparatus. — Cages, Feed, etc., Abderhalden, 5, 1035; Tigerstedt, 1.3, i.
Tread-mill, Abderhalden, 3, 1050.
Dogs (and Feed). — Abderhalden, 3, 1041; Pratt, 1909, Feeding, Amer. Jour. Physiol.,
24, 269.
Ruminants. — Abderhalden, 3, 1054; collection excreta, ibid., 6, 453.
Mice, Guinea-pigs, etc. — Feeding, Abderhalden, 3, 1269.
Rats. — Food and Growth, Osborne and Mendel, 1913, Jour. Biol., Chem., 15, 311;
Street, 1915, Jour. Amer. Med. Assoc., 64, 638.
Suine. — Metabolism cage, Forbes, 1915, Ohio Agr. Exp. Sta., Cir. 152.
Meat, Protein Content, Bedford and Jackson, 1916, Proc. Soc. Exp. Biol. Med., 13, 83.
Feces. — Delimitation, Abderhalden, 3, 999; 5, 333. (Charcoal, 0.2 gm., or carmin, 0.3
gm. in capsule before breakfast appears in stool of next morning.) Examination of human,
Abderhalden, 5, 331. Examination of herbivorous, ibid., 3, 263. Preservation, Howe,
Rutherford, and Hawk, 1910, Jour. Amer. Chem. Soc., 32, 1683. Fat, Abderhalden, 5,
363; Saxon, 1914, Jour. Biol. Chem., 17, No. 2. Bacteria, Abderhalden, 5, 359. Ash,
ibid., 5, 331.
Urine. — Collection, human, Abderhalden, 3, 999; 5, 281 ; pan for quantitative collection
of female urine, Folinvand Denis, 1915, Arch. Int. Med., 16, 195. Preservation, ibid., 5,
283; Gill and Grindley, 1909 (Thymol and cold), Jour. Ajmer. Chem. Soc., 31, 695. Gen-
eral Urine Analysis, Abderhalden, 3, 765; 5, 281.
Nitrogen. — Folin, 1915, Jour. Biol. Chem., 21, 195; Bock and Benedict, 1915, ibid., 20
\<>. i ; Gradwohl, 1916, Jour. Amer. Med. Assoc., 67, 809.
Urease, Dunning, 1916, Amer. Jour. Phar., 5, 809; for Urea in Urine, Fiske, 1916,
Jour. Biol. Chem., 23, 455; in blood, Marshall, 1913, ibid., 14, 283; 15, 487.
A mi no-nitrogen. — Van Slyke, 1915, Soc. Exp. Biol. Med., 13, 63.
Pur in Bases. — Graves and Rober, 1915 (nephelometric), Proc. Amer. Soc. Biol. Chem.
3, i?.
:n and Creatinin. — Morris, 1915, ibid., 3, 15; Janney and Blatherwick, 1915,
Jour. Biol. Chem., 21, 567 (in muscle and organs).
Phenols in Urine and Feces. — Folin and Denis, 1916, Jour. Biol. Chem., 26, 507.
Bence-Jones Proteinuria. — Folin and Denis, 1914, Jour. Biol. Chem., 18, 277.
Blood-gas Analysis. — Barcroft, 1914, "Respiratory Function of Blood."
Carbon Dioxid, Tension in Alveolar Air. — Marriott, 1916, Jour. Amer. Med. Assoc.,
66, 1594; Combining Pou'er of Plasma, Van Slyke, Stillman, and Cullen, 1915, Soc. Exp.
Biol. Med., 13, 39.
Birds. — Abderhalden, 3, 1058; Anesthesia, operations, urine secretion, Sharpe, 1912,
Jour. Physiol., 31, 75.
-Urine Collation. Denis, 1912, Jour. Biol. Chem., 13, 225. General Experi-
ments, Fiu-lincr, 52; Operative Teehnie. Alxk-rhalden, 3, 1103; Isolated heart, Beresin, 1913,
Anh. get. I'liy-i'.l.. 150, 549.
nr Animal*.— Al>drrh;ildrn. J, 1064.
' organisms.— Ahdi-rlialdon, 5, 1158.
Perfusion. — Ibid., 5, 1245.
Surviving Organs. — Bagttoni in Abderhalden, 3, 358.
(D) CENTRAL DEPRESSANTS
Introduction.— The effects of central depressants, as seen in mammal-.
are not a> sharply locali/.cd as in fro^s. Tin- symptoms usually U^in with
stupidity and <ln>\\ ^incss, with or without CM itcniciit ; ataxia. -Ircp. and
i. The respiration i- usually slowed, more than corresponds to the
muscular <|\iict. Thr rrllexes are usually diminished, but with morphin
they may he increased. The temperature tends to fall. Tin- detail- of
these actions determine^ their practical availability as analgesics, hypn.
or anesthetics.
228 A LABORATORY GUIDE IN PHARMACOLOGY
Depression of the brain interferes in the first place with the higher psychic processes;
this passes into sleep, and finally into anesthesia. Depression of the spinal cord leads to
loss of reflex excitability; depression of the medulla, to fall of blood-pressure, quickening of
the pulse, slowing of respiration, and fall of temperature. The location of the action of
the depressants is therefore indicated by the symptoms.
The readiness with which the successive stages may be produced and their duration
varies with each drug, and determines its uses in therapeutics.
Those which act mainly on the higher centers are used for the relief of pain (analgesics)
or for producing sleep (hypnotics) : Morphin, Cannabis, Alcohol, Chloral.
Those which act profoundly on the brain and spinal cord are employed for operative
anesthesia (general anesthetics) : Ether, Chloroform, Ethyl Chlorid, etc.
Paralysis of the medulla (Chloral, Chloroform) is only utilized in experimental technic;
but it is important as a source of danger in anesthesia. It is treated mainly by artificial
respiration.
It will be remembered that most central stimulants also produce some depression;
similarly, the depressants often cause some stimulation. Morphin may produce excite-
ment in certain individuals; it may also stimulate the vomiting and defecating and motor
centers. It always increases the reflex excitability of the spinal cord, and may even cause
typical strychnin spasms in the lower animals.
Alcohol and the general anesthetics produce a preliminary stimulation. This, how-
ever, is not due to a direct stimulant action, but to inhibition of restraining centers and to
reflex stimulation.
Observations. — In observing the effects, attention should be directed
especially to the general behavior of the animal (excitement, drowsiness,
ataxia, sleep, coma, etc.); to the respiration, pulse, and temperature; the
reflexes (patellar, ear, etc.) ; the pain reaction (sudden and gradual pressure
on foot), etc. These should be observed before and at intervals after the
administration. Care must be taken that the animal is not excited when the
normal observations are taken. If the respiration becomes irregular,
endeavor to obtain tracings (with a lever attached by a bulldog clamp to
the hair of the chest or abdomen) .
TECHNICAL REFERENCES
Central Depressants. — Robert, Intox., i, 223.
Psychologic Tests.— Tigerstedt, 3.5; Mental tests, Dana, 1913, Med. Rec., Jan. 4;
Binet Scale, Pop. Sci. Mo., Jan., 1914.
EXERCISE XIII.— MORPHIN
(REPORTER IV, A)
Experiment i. (Group II, A) Dog. — Inject hypodermically 10 mg. (i c.c.
of 4 per cent.) per kg. and carefully observe the effects. The animal will
probably vomit and pass feces and sometimes urine (stimulation of
medullary and spinal centers). The respiration may be temporarily
quickened, but will soon become slowed and more shallow (stimulation
and depression of the respiratory center). A tracing may be taken. The
pulse-rate will decrease (stimulation of vagus center). The temperature
falls (general lowering of metabolism). The pupils are variable (central
action). The animal becomes more quiet; does not move spontaneously,
and the movements are shivering. The hind legs are especially affected,
and may be dragged when the animal walks. The dog does not usually
fall asleep, but pain is felt less acutely. The reflexes, however, are not
diminished.
The effect is, on the whole, a central depression; the action differs from
that on man mainly by the absence of sleep, and by the presence of the diar-
rhea, by the variability of the pupils, and by the more pronounced motor
disturbances.
CHAP, xxxix METABOLISM; DEPRESSANTS; IRRITANTS 229
Experiment 2. (Group II, B) Cat. — Inject hypodermically 20 mg. (J
c.c. of 4 per cent.) per kg. The effect may be excitant, the animal running
about; the pupils dilate; however, analgesia is present.
Experiment 3. (Group II, A) Rabbit. — See Exercise II.
Questions. — (a) Describe the effects of morphin on the three animals.
(b) Which is most and which least susceptible to the narcotic action
(considering the dosage)?
(c) What are the most conspicuous qualitative differences in the actions?
Experiment 4. (Demonstration) Mouse Test for Morphin. — The hypo-
dermic injection of morphin into white mice is followed in two to twenty
minutes by a peculiar position of the tail, which is carried in a rigid,
usually s curve over the back. This is maintained for one or two hours.
The reaction is characteristic for morphin (above o.oi mg. for mouse of
15 to 20 gm.) ; it is also given by some of the other opium alkaloids and apo-
morphin.
Inject under back of white mouse morphin 0.5 mg. (0.5 c.c. of i : 1000)
and observe results.
Technical References.— Straub, 1911, Deut. med. Woch., 37, 1462;
Fuehner, Nachweiss, 150.
Experiment 5. (Optional) Synergism of Opium Alkaloids. (See W. Straub, 1912,
Bioch. Zs., 41, 4191-) — Inject hypodermically into white mice the following drugs, and
note whether they survive or die. The dosage refers to mice of 15 to 20 gm. :
(1) Morphin, 12 mg.
(2) Morphin, 18 mg.
(3) Narcotin, 10 mg.
(4) Narcotin, 2 mg., with morphin, 2 mg.
(5) Narcotin, 4 mg., with morphin, 4 mg.
(i), (3), and (4) should survive; (2) and (5) should die.
Questions. — (a) What effect has narcotin on the toxicity of morphin? (Compare i
and 2 with 3 and 4.)
(6) Is this a simple addition of the toxicity of the two drugs?
(c) What is this action called:'
Experiment 6. (Optional) Papaverin. — Inject cat, hypodermically, with 100 mg. per
kg.: narcosis.
Experiment 7. (Optional) Synergism of Morphin, Scopolamin, and Atropin, Cat. —
three cats hypodermically with morphin, each 20 mg. (i c.c. of 4 per cent.) per kjr.
Use Cat A. as control. Into Cat B inject Scopolamin, 0.5 mg. per kg. ($ c.c. of i : 1000);
into Cut (' injr. t Atropin, i mg. per kg. (i c.c. of i : 1000). Inject Cat D with Scopolamin
and Cat K with Atropin, using the same doses, both without Morphin. Compare the
rr>ults.
EXERCISE XIV.— CANNABIS
(REPORTER IV, A)
The administration of cannabis to dogs usually produces vomiting and
some excitement. In one or two hours this is followed by muscular in-
coordination (ataxia), and, finally, by lassitude, depression, and sleep.
Tin- individual susceptibility varies. Small, short-haired dogs (fox terrier- >
are nn»>t Miitable. The effects do not occur on hypodermic administration.
The activity is due to resinous constituents.
Experiment i. (Group III, B) Effects.— Administer a capsule containing
.ict of Cannabis Indie a 0.05 gni. per ktf. Thi> is done by drawing out
the tongue and placing the rap-tile l.aik a- far U po— il>le. On ;
the tongue the < apMile i- usually >\vallowed easily. If not. the- mouth i>
held >hut and the animal slapped on the throat. Observe the effects of
the cannal»i-
Questions. — (a) Describe ihe d the cannabis.
(b) Why i> it inattive hvpodennu ally?
230 A LABORATORY GUIDE IN PHARMACOLOGY
Experiment 2. (Optional) Bio-assay of Cannabis. — This is performed similarly to
Experiment i. Details, U. S. P. IX. The standard dose for producing muscular inco-
ordination in dogs is per kg.: fluidextract, 0.03 c.c.; extract, 0.004 gm.
Technical References. — U. S. P. IX; Pittenger, 98; Jour. Amer. Pharm. Assoc. (Com-
mittee), i, 1305, 1912; Houghton, 1911, Amer. Pharm. Assoc. Bui., 6, 176.
EXERCISE XV.— (DEMONSTRATION) MAGNESIUM AND CALCIUM
(REPORTER IV, A)
Magnesium produces a depressant action, with sensory and motor
paralysis, both central and peripheral. Calcium is also depressant, but
nevertheless it antagonizes the magnesium effects, so that the animal re-
covers immediately.
A rabbit has received intramuscularly magnesium sulphate (crystals),
1.75 gm. (7 c.c. of 25 per cent.) per kg. When paralysis is complete, 6 to
8 c.c. of 3 per cent, calcium chlorid is injected slowly into the jugular vein:
immediate recovery.1
QUESTIONS
(a) Describe the effects of magnesium.
(b) Could this be utilized clinically for anesthesia?
(c) Describe the effects of calcium on the magnesium rabbit.
(d) Explain the antagonism.
TECHNICAL REFERENCE
Meltzer and Auer, 1907, Soc. Exp. Biol. Med., 5, 33.
EXERCISE XVI.— (GROUPS III AND IV) ALCOHOL AND TREATMENT OF
ALCOHOL POISONING
(REPORTER IV, A)
Record respiration, temperature, and general symptoms (Pilcher, 1912,
Jour. Pharmacol., 3, 267). Cats are used.
Experiment i. (Group III, A) Alcohol Control. — Inject into cat by stom-
ach-tube Alcohol 4 c.c. (16 c.c. of 25 per cent.) per kg. Observe symptoms
and course for control.
Experiment 2. (Group III, B) Alcohol and Emesis. — Inject Alcohol as in
Experiment i. When symptoms are fully developed, or in about one-half
hour, administer Zinc Sulphate, 25 c.c. of i per cent., by stomach-tube.
Compare course with Experiment i.
(If one of the cats should have vomited spontaneously, it will not be
necessary to administer the emetic.)
Experiment 3. (Group IV, A) Alcohol-caffein Antagonism. — Inject
Alcohol as in Experiment i. When symptoms have fully developed, or in
about one-half hour, inject, hypodermically, Caffein, 20 mg. (2 c.c. of i per
cent.) per kg. Observe immediate effect and compare subsequent course
^7ith Experiment i.
Experiment 4. (Group IV, B) Alcohol-caffein Synergism. — Inject
Alcohol as in Experiment i. Follow this at once with a hypodermic injec-
tion of Caffein, 50 mg. (5 c.c. of i per cent.) per kg. Compare course with
Experiment i.
» This rabbit may then be used for Exercise II.
CHAP, xxxix METABOLISM; DEPRESSANTS; IRRITANTS 231
QUESTIONS
(a) Describe the phenomena of alcohol poisoning.
(b) Name two methods of treatment and compare their efficiency as to
immediate and ultimate improvement.
(c) Does the antidotal efficiency of caffein increase with the dose?
Explain.
EXERCISE XVn.— (GROUP I) CHLORAL POISONING AND TREATMENT
(REPORTER IV, A)
Chloral is a typical depressant. Cats are used. The effects increase
with the dosage as follows (the dosage refers to gm. per kg., administered
as 2.5 per cent, solution by stomach-tube to cats):
0.09 to 0.15: natural sleep.
o.i 8 to 0.25: light coma; recovery over night.
0.3 and higher: deep coma; recovery in one to four days.
0.35 to 0.50 (mean, 0.44): fatal.
Unless vomiting occurs (which is not infrequent), 0.5 gm. per kg. may
be accepted as surely fatal.
Observe drowsiness; equilibrium; pain; reflexes; respiration; pupils;
temperature.
Technical References. — Sollmann and Hatcher, 1908, Jour. Amer. Med.
Assoc., 51, 487.
Experiment i. (Group I, A) Symptoms of Chloral Poisoning. — See
Exercise I. Administer by stomach-tube a fatal dose of chloral, 0.5 gm.
(20 c.c. of 2.5 per cent.) per kg.
Experiment 2. (Group I, B) Chloral and Heat. — Proceed as in Experi-
ment i, but keep the animal warm. Compare the results.
Experiment 3. (Group I, A) Chloral and Caffein. — Inject Chloral as in
Experiment i. Fifteen minutes later give Caffein, 10 mg. (i c.c. of i per
cent.) per kg., hypodermically. Compare immediate and ultimate results.
Experiment 4. (Group I, B) Chloral and Strychnin. — Inject Chloral as
in Kxperiment i. After fifteen minutes begin treatment with Strychnin:
administer o.i mg. (o.i c.c. of i : 1000) per kg., hypodermically, and re-
peat every half-hour unless the animal becomes spasmodic. Compare
immediate and ultimate results.
Experiment 5. (Optional) Chloral and Antidotes in Rabbits. Administer to rabbits,
by Stomach-tube, Chloral. 0.5 gm. prr kg. \Yhrn light narcosis has >i-t in, try tin- follow-
ing drug-, I iy vein (doses arr prr kg.): immrdiatr n-vival with Corain. 5 mg. (may IK
01040 nig.; beta trtra hvdronaphthylamin, loto 20 mg.
No revival: IMu-n.il (hut twitdiings); Epim-phrin or Pituitary (Y. Airila, 1913, Arch.
Int. Pharmacod., 23, 453).
QUESTIONS
(a) Describe the effects of t hloral.
(b) How may these be treated?
(c) How do these methods compare in efficient
(d) Are all the symptoms relieved to the same degree?
(e) Suggest other methods of treatment.
(E) GASTRO-ENTERITIS
Introduction. The most important phenomena of poisoning by irritants
are caused by gastro-rnlenii^. The principal symptoms consist in very
severe abdominal pain; prol'u-e vomiting and diarrhea; and reflex collapse.
232 A LABORATORY GUIDE IN PHARMACOLOGY
If the irritant is also corrosive, the discharges are bloody or otherwise dis-
colored. The stools are generally very watery.
Observations Required. — Keep in cage and collect urine. During life
note the vomiting, watery diarrhea, and general depression. At autopsy
note the congestion of the abdominal organs, particularly the mucosa of
the alimentary canal. Observe the character of the contents, and look for
corrosions. Corrosions are most pronounced in the case of the mercury;
they are absent with colchicum. The latter causes intense congestion in
ridges. Observe that the arsenic produces its effects, even when it is given
hypodermically (note particularly the fluid contents). The animal usually
lives several hours or longer. The urine of mercury and arsenic will gen-
erally contain albumin and casts. The mercury and veratrin animals
may recover, but will show erosion of the stomach on autopsy.
As these experiments would be painful, they are to be performed on the
morphinized animals of Exercise XIII.
EXERCISE XVm.— PHENOMENA OF CASTRO-ENTERITIS
(REPORTER V, A)
Experiment i. (Group II, A) Colchicum. — Administer by stomach- tube
to morphinized dog (or cat) Fluidextract of Colchicum 0.5 c.c. per kg.: no
symptoms for several hours; but on the next day the animal will be found
dead, with evidence of bloody diarrhea and hemorrhagic congestion of
intestines. Autopsy.
The alkaloid of colchicum is practically inactive, but is converted in
the tissues of mammals into oxydicolchicin, which is the toxic agent. This
explains the long interval between administration and symptoms. The
drug is not at all corrosive. It has been suggested that it does not irritate
directly, but that it merely exaggerates the normal irritability of the
intestine.
Experiment 2. (Group II, B) Mercuric Chlorid. — Inject by stomach- tube
into morphinized cat Mercuric Chlorid 5 mg. (5 c.c. of i : 1000) per kg.
Notice the white (cooked) appearance and hardness of the gastric
mucosa at the autopsy.
Experiment 3. (Group II, A) Arsenic. — Inject hypodermically into
morphinized rabbit Sodium Arsenate, 50 mg. (i c.c. of 5 per cent.) per kg.
The symptoms and lesions of arsenic poisoning bear the closest resem-
blance to those of local inflammation of the alimentary tract. It can be
shown, however, that the direct irritant or corrosive action of the poison is
entirely inadequate to produce this inflammation, especially when the poison
is given hypodermically or intravenously. Its action is really due to
direct paralysis of the capillaries, with increased permeability. This is
also the main phenomenon of inflammation. The lesions are therefore
identical. A characteristic clinical feature of acute arsenic poisoning
consists in the "rice-water" stools, which consist of a profuse watery exudate
with shreds of desquamated mucosa.
QUESTIONS
Describe the symptoms and lesions of poisoning by colchicum, mer-
curic chlorid, and arsenic.
Experiment 4. (Optional) Veratrin. — i c.c. of i per cent, by stomach, rabbit. Vera-
trin is one of the very few alkaloids which are directly corrosive.
CHAP, xxxrx METABOLISM; DEPRESSANTS; IRRITANTS 233
TECHNICAL REFERENCES
Experimental Hepatic Cirrhosis. — Methods are described by: Pearce, 1906, Jour.
Exp. Med., January; Opie, 1910, Trans. Assoc. Amer. Physicians, 25; 1912, ibid., 117;
Grover, 1913, Jour. Amer. Med. Assoc., 61, 458; Lissauer, 1914, Arch. Path. (Virch.), 217,
56. •
Spontaneous Hepatic Cirrhosis of Rabbits. — Grover, 1915, Jour. Amer. Med. Assoc.,
64, 1487-
Fatty Degeneration of Liver. — Phosphorus, Abderhalden, 5, 1232.
EXERCISE XIX.— (OPTIONAL) MORPHIN ON COLOCYNTH DIARRHEA
Administer to cat, by stomach-tube, 10 c.c. of a 10 per cent, infusion of Colocynth.
After two hours decerebrate and expose intestines. They should be in violent peristalsis.
Inject hypodermically Morphin, 20 mg.: the peristalsis should be promptly arrested
(Padtberg, 1911, Arch. ges. Physiol., 139, 318; Takahashi, 1915, ibid., 159, 327).
(F) NEPHRITIS
Introduction. — The action of irritants is proportional to their concentra-
tion. This is greatest where they enter and leave the body — in the alimen-
tary canal and in the kidneys. During their passage through the body they
are generally diluted to such a degree that the irritation of other tissues
is seen only when they are administered continuously. It may then lead
to increased formation of fibrous tissue (arteriosclerosis and cirrhosis).
Nephritis, however, often occurs acutely and is produced by all absorbable
irritants.
Rabbits can be conveniently used for the production of experimental
nephritis. The presence of albumin, casts, and sugar should be sought
for in the urine, and the kidneys should be hardened, stained, and examined
histologically.
EXERCISE XX.— (DEMONSTRATION) URANIUM HYDROPS
(REPORTER V, A)
Inject hypodermically into rabbit 5 mg. (i c.c. of 5 : 1000) of Uranium
Nitrate. Repeat daily for three days.
EXERCISE XXI.— (OPTIONAL) OTHER NEPHRITIC POISONS
Arsenic— Mainly Glomerules. — Inject hypodermically 10 mg. per k<^.
of Potassium Arsenate: the urine becomes albuminous in ten min
The glomeruli are dilated, filling Bowman's capsule. The epithelium of
the convoluted tubules is affected to a varying degree; the straight tubules
are not involved.
Aloin — Mainly Epithelium of Convoluted Tubules. — Inject hypoder-
mic ally 2 c.c. per kg. of a 5 per cent, solution; repeat for two or three days.
The action is practically limited to the convoluted tubules.
Chromates As Aloin. -Inject hypodcrmically 30 mg. per kg. of Potas-
sium Birhmmatc: nephritis is plain in twenty-four 1m
Cantharidin— All Renal Elements.— Inject hvpo.lermically 5 mg. per
<lissolved in acetic et lu-n: albuminuria in ten minutes. ,
Mercuric Chlorid— Mainly Interstitial. —Inject hypodcrmically 10 c.c.
of i : 1000 solution daily: albuminuria in two to three days.
Oxalates Occlusion of Tubules by Crystals of Calcium Oxalate. — Inject
hypodermically 0.250^111. <>f Ammonium (Kaiate into a rabbit.
Chloroform, Phosphorus, Hydrazin. Fi^ke and Karsncr, 1914, Jour.
Biol. Chem., 18, 381.
234 A LABORATORY GUIDE IN PHARMACOLOGY
TECHNICAL REFERENCES
Production of Experimental Acute Nephritis. — Pearce, Harvey Lect., 1910; Sollmann,
1904, Jour. Amer. Med. Assoc., Nov. 26; MacNider, 1912, Jour. Med. Res., 26, 79.
Unilateral Nephritis. — Quinby and Fitz, 1915, Arch. Int. Med., 15, 303.
Experimental Chronic Nephritis. — Emerson, 1908, Arch. Int. Med.; Opie, 1912, Trans.
Assoc. Amer. Physicians, 27, 117; O'Hare, 1913, Arch. Int. Med., 12, 49; Karsner and
Denis, 1914, Jour. Exp. Med., 19, 270.
Renal Circulation in Nephritis. — Schlayer and Hedinger, 1907, Deut. Arch. klin. Med.,
90, i.
Protein in Urine. — Quantitative, Folin and Denis, 1914, Jour. Biol. Chem., 18, 273;
Quantitative Estimation by Biuret Reaction, Autenrieth and Mink, 1915, Muench.
med. Woch., 62, 1417; Comparison of Clinical Methods, Kahn and Silberman, 1914, N.
V. Mod. Jour.. Oct. 3; Comparison of Gravimetric and Nephelometer Methods, Mar-
shall, Banks, and Graves, 1916, Arch. Int. Med., 18, 250.
Plasma Proteins. — Quantitative Estimation, Cullen and Van Slyke, 1916, Proc. Soc.
Exp. Biol. Med., 13, 197.
Elastometer for Measuring Edemas. — Schade, 1912, Zs. Exp. Path. Then, n, 369;
A. B. Schwartz, Arch. Int. Med., 17, 396; Maver and Schwartz, ibid., 459.
(G) REFLEX EFFECTS OF IRRITANTS; ARSENIC ON CIRCULATION
Sensory reflexes produce marked changes in the circulation and respira-
tion. The effects differ for each region, but are naturally least marked
where sensation is least developed, i. e., in the gastro-intestinal tract.
EXERCISE XXII.— (OPTIONAL) TRIGEMINAL— VAGUS (KRETSCHMER)
REFLEX— CHLOROFORM, AMMONIA
Feel pulse of rabbit. Blow into nostrils the vapor of chloroform; and
when the animal has recovered, the vapor of ammonia: marked slowing or
temporary arrest of the heart.
EXERCISE XXIII.— (DEMONSTRATION) IRRITANTS ON BLOOD-PRESSURE
AND RESPIRATION
(REPORTER V, A)
Morphinize dog (20 mg. =| c.c. of 4 per cent, per kg.). Etherize. Insert
tracheal cannula with T piece and connect with tambour for respiratory
tracing. Connect carotid artery for blood-pressure tracing; femoral vein
for injection. Remove ether. Start slow tracings.
Experiment i. Tracheal Irritation. — Blow ammonia vapor into trachea:
little or no effect.
Experiment 2. Laryngeal Irritation. — Blow ammonia vapor into mouth
so as to reach larynx : marked disturbance of respiration and blood-pressure.
Experiment 3. Irritants in Mouth.— With a pipet flood the mouth with
5 per cent, acetic acid: marked disturbance (mainly from larynx).
Experiment 4. Corrosives in Stomach, Intestines, and Peritoneum. —
Make small opening into abdomen, expose stomach and intestines, and with
a pipet apply concentrated nitric acid successively to the interior of the
stomach and intestines, and to the visceral and parietal peritoneum: usually
but little effect.
EXERCISE XXIV.— (DEMONSTRATION) ARSENIC ON CIRCULATION
(REPORTER V, A)
Inject intravenously 50 mg. per kg. of Arsenate of Sodium (i c.c. per kg.
of 5 per cent.) : the intestines show capillary congestion and become filled
with fluid (paralysis of the capillary walls). The blood-pressure falls, but
CHAP. XL CONVULSANTS AND TREATMENT OF POISONING 235
rises at once if the aorta is temporarily compressed, showing that the cardiac
muscle is not injured (except by larger doses). Stimulation of the sciatic
or splanchnic nerve also causes a rise. Kill the animal and examine the
gastro-intestinal lesions.
QUESTIONS
(a) Which surfaces give the most, which the least, reflexes with irritants?
(b) What therapeutic use could be made of these reflexes?
(c) How can the danger of reflex arrest of the heart by chloroform in-
halation be minimized?
(d) What effects has arsenic on the circulation?
(e) Is the effect due primarily to depression of the heart? Of the vaso-
motor center? What then?
(/) Describe the autopsy lesions of arsenic,
(g) Describe the autopsy lesions of nitric acid.
TECHNICAL REFERENCES
Irritant Reflexes. — Heidenhain and Gruetzner, 1877, Arch. ges. Physiol., 16, 55;
Sollmann, 1907, Amer. Jour. Physiol., 20, 74.
CHAPTER XL
CONVULSANTS AND TREATMENT OF POISONING
(REPORTER III, D)
(A) CONVULSANTS
Introduction. — The effects of convulsant poisons are very similar in
frogs and in mammals. They can be localized by the same methods,
but the technic is naturally more difficult in the higher animals. Only the
symptoms will be studied in this exercise. The seat of the action is the same
as in the frog.
Spinal conwdsants produce increased reflex excitability, and then
tetanic opisthotonus. Strychnin is the principal example; caffein belongs
to the same group.
Medullary conmdsants produce clonic spasms with tendency to empros-
t h<>t onus. Nicotin and hydrocyanic acid belong to this group. They
act by producing asphyxia, which is the direct cause of the convulsions.
Veratrin, camphor, picrotoxin, ammonium, and some others act directly
on tin- (vi
Cerebral conmdsants act on tin- motor areas. They produce rhythmic
twitchings of muscles (chorciform contractions) or epilcptiform spa-m-.
These are sometimes seen in morphin poisoning. They are also produced
by absinth.
produce constant motion, but of a purposive type.
plainly due to excitement. The movements may as-unie various types:
there may be Dimply an iiirrea ity. as with atropin; or the animal
l>ecome maniacal, as sometimes with cannabis; or it may run constantly
in a circle.
The iriitral action may aUo remain localized in certain (It-finite «
Small doses of caffein, for instance, cause an increase of psychic activity
236 A LABORATORY GUIDE IN PHARMACOLOGY
and tendency to wakefulness. Apomorphin acts mainly on the vomiting
center; the antipyretics on the temperature. Drugs may also stimulate
the vasomotor, vagus, or respiratory center, etc.
The action of convulsants on mammals is often not sharply localized,
but involves different centers in succession, generally from the brain down-
ward. Cocain, phenol, and asphyxia are examples.
It will be noted, in the following experiments, that the stimulation is
generally followed by depression.
TECHNICAL REFERENCES
Stimulation of Motor Areas, Stewart, 962; Tigerstedt, 3.4, 107; Operations on Brain,
ibid., 79.
OBSERVATIONS
Observe the respiration, general behavior, reflexes, and the onset and
type of the convulsions, and time of death.
EXERCISE I.— (GROUP H, A) STRYCHNIN HYPODERMICALLY
(Spinal convulsions.) Administer hypodermically to cat a fatal dose of
strychnin, 0.75 mg. (f c.c. of i : 1000) per kg.: increased reflexes, increased
respiration; convulsions, first on stimulation, soon spontaneously; sym-
metric, first clonic, then tetanic. Respiration arrested during spasms by
fixation of muscles; asphyxial symptoms: dilated pupils, cyanosis. Depres-
sion between convulsions. Convulsions start in from fifteen minutes to
one and one-half hours. Death occurs in from thirty minutes to three;
hours. Make a sketch-drawing of the tetanic animal.
EXERCISE H.— (GROUP HI, B) STRYCHNIN BY STOMACH
Administer by stomach-tube to cat a fatal dose of Strychnin, i mg..
(i c.c. of i : 1000) per kg. : effects as in Exercise I, but usually rather slower..
QUESTIONS
(a) Describe the course of strychnin poisoning.
(b) Is there as much difference in the toxicity, by stomach and hypoder-
mically, as was observed with rabbits (Chapter XXXVIII)?
EXERCISE HI.— (GROUP I, A) CAMPHOR CONVULSIONS (CEREBRAL AND
MEDULLARY)
Experiment i. — Administer by stomach- tube to cat or rabbit Camphor >
2 gm (10 c.c. of 20 per cent, in oil) per kg.: convulsions occur in about
half an hour or later. They are Violent, but asymmetric and irregular.
Try whether they can be controlled by inhalation of chloroform. They
usually run a long course.
Questions. — How do camphor convulsions differ from those of strychnin?
Experiment 2. (Optional) Camphor Toxicity Modified by Method of Administration. —
In guinea-pigs dry camphor by mouth is fatal with a dose of 0.14 to 0.18 gm. per 100 gm.
It is less toxic when dissolved in oil. Hypodermically, an oily solution is also less toxic
than a solution in alcohol or water; but the oily solution is more toxic hypodermically than
by mouth. Peritoneal injection is more toxic than hypodermic, the oily again being the.
least effective (Cairis, 1914, Jour. Pharm. Chem., 10, 224).
CHAP. XL CONVULSANTS AND TREATMENT OF POISONING 237
EXERCISE IV.— (GROUP I, B) TREATMENT OF EPILEPTOID (CAMPHOR)
CONVULSIONS BY BROMID
(Adapted from Januschke and Inaba, 1913, Zs. exp. Med., i, 129.)
On morning of previous day administer to cat or rabbit by stomach-
tube Sodium Bromid, 2 gm. per kg. (10 c.c. per kg. of 20 per cent.). Repeat
at six-hour intervals, giving the last dose an hour before the laboratory
period. Administer Camphor to this bromid-cat, as in Exercise III. Com-
pare the results. Calcium also suppresses the convulsions (Januschke
and Hirsch, 1913, Ther. Mon., 27, 777).
QUESTIONS
(a) Describe the bromid symptoms.
(b) Record the camphor results.
(c) How does the bromid suppress the epileptic convulsions?
EXERCISE V.— (OPTIONAL)
Experiment i. Veratrin (Stimulation of Medulla). — Inject hypodermically into a
rabbit i mg. per k<j. of \\ratrin salt (i c.c. per kg. of ^ per cent.). Repeat in twenty
minutes, if necessary: salivation, inco-ordination, irregular convulsions, animal jumps
ht up ("bucks"). Paralytic condition. If death should occur, the respiration stops
before the heart. (The commercial samples of veratrin vary considerably in their ac-
tivity, and it may therefore be difficult to hit upon the proper dose which is required to
produce the "bucking.")
Experiment 2. Absinthe (Epileptic Cerebral Convulsions). — Inject 0.03 to 0.05 of
Absinthe Essence per kg.
(B) TREATMENT OF POISONING
Introduction. — The main features of the treatment of poisoning consists
in:
(1) Chemic precipitation, neutralization, or destruction of the poison.
(2) Removal of the poison.
(3) Physiologic antidotes.
(4) General supporting measures.
All treatment must be as prompt as possible.
(1) Chemic Antidotes.— These have been discussed in Chapter XVI,
\\hich should be consulted.
(2) Removal of the Poison. — This is accomplished by washing, emesis,
lavage, catharsis, and diuresis.
(3) Physiologic Antidotes. — The effects of depressant drugs are counter-
acted by stimulants, and vice vena. It must be remembered, ho\ve\cr.
that the action of stimulants passes readily into depression, which would
increase the danger. Antidotes should therefore be given in rather moderate
doses. It should also be borne in mind that physiologic antidotes ran
only the -\ mptoms, and not the action of the poison. They are therefore
useful only when the symptoms are a direct source of danger. In the case
•ychnin, for instance, death is due to the direct depressant ai tion of the
drug, aided by the exhaustion consequent on the convulsions. Chloral,
re, or artificial respiration, by preventing the omvul>ion-. are able to
save an anima era 1 1 imes t he fatal dose, but they are quite ineffec t i\ e
against doses sufficiently large to kill by the direct depressant action of the
poison.
(4) General Supporting Measures. — The immediate cause of death
with most poisons consists in failure of the respiration. This should
238 A LABORATORY GUIDE IN PHARMACOLOGY
be carefully watched and supported by hot coffee. Should this prove
insufficient, artificial respiration must be instituted, and this before the
natural respiration has ceased. The patient should be kept warm. Pain
(from corrosives, etc.) should be controlled by morphin or the local use of
cocain.
The use of antidotes is well illustrated by Strychnin, as in the following
exercises.
TECHNICAL REFERENCES
Saline Infusion on Excretion of Toxic Substances. — Lenhartz, 1899, Deut. Arch. Klin.
Med., 64, 189.
Vividiffusion. — Abel, Rowntree, and Turner, 1914, Jour. Pharmacol., 5, 275; MacCal-
lum and Lambert, 1914, Soc. Exp. Biol. Med., n, 78.
EXERCISE VI.— CHEMIC ANTIDOTES
Experiment i. (Group IV, B) Strychnin and Permanganate. — Admin-
ister Strychnin by stomach-tube as in Exercise II. Follow within five
minutes by Potassium Permanganate, 15 c.c. of i per cent, per kg. Com-
pare the results.
Experiment 2. (Group V, B) Hydrocyanic Acid and Permanganate. —
Administer to cat, by stomach-tube, Hydrocyanic Acid, 2 mg. (2 c.c. of
i : 1000) per kg. (twice fatal dose). Follow this at once with Potassium
Permanganate, 15 c.c. of i per cent, per kg. The animal usually shows
severe symptoms, but survives.
QUESTIONS
(a) Report the results.
(b) What is the mechanism of the action of permanganate?
(c) What would interfere with its usefulness?
EXERCISE VII.— (GROUP V, A) ADSORBENT ANTIDOTES (STRYCHNIN
AND CHARCOAL OR CARAMEL)
Administer Strychnin by stomach- tube as in Exercise II. Follow at once
with a suspension of 25 gm. of Charcoal or of 25 gm. of Caramel. Compare
the results with Exercise II.
QUESTIONS
(a) Report the results.
(b) How do the charcoal and caramel act?
(c) How could their efficiency be increased?
TECHNICAL REFERENCE
Charcoal as Antidote.— O. Adler, 1912, Wien. Klin. Woch., 25, No. 21.
EXERCISE VHL— (GROUP IV, A) EVACUATION (STRYCHNIN AND LAVAGE)
Administer Strychnin by stomach-tube as in Exercise II. Five or ten
minutes later wash the stomach. Compare the results.
QUESTIONS
(a) Report the results.
(b) Would lavage be of much use after convulsions have set in?
CHAP. XLI RESPIRATION (AND BLOOD-PRESSURE) 239
EXERCISE IX.— (GROUP H, B) ARTIFICIAL RESPIRATION AND STRYCHNIN
Administer Strychnin hypodermically as in Exercise I. When the animal
becomes convulsive, start artificial respiration. The convulsions are sup-
pressed. Note that they return if the respiration is intermitted. Continue
the respiration until the animal is out of danger. Compare the results with
Exercise I.
QUESTIONS
(a) Describe the effect of artificial respiration on strychnin poisoning.
(b) Explain the effect.
(c) Should the artificial respiration be applied only during the convul-
sions, or how?
EXERCISE X.— (GROUP HI, A) PHYSIOLOGIC ANTIDOTE (STRYCHNIN
AND CHLORAL)
Administer Strychnin hypodermically as in Exercise I. Follow this at
once by Chloral, 0.25 gm. (10 c.c. of 2.5 per cent.) per kg., by stomach-tube.
(This dose produces light coma in normal animals.) Compare the results
with Exercise I.
QUESTIONS
(a) Describe the results.
(b) How does the chloral act as antidote?
(c) Would it be useful in other convulsions?
CHAPTER XLI
RESPIRATION (AND BLOOD-PRESSURE)
Introduction. — The respiratory center may be stimulated or depressed
by the direct or reflex action of drugs; or indirectly, for instance, by changes
in the circulation, by acidosis, etc. The respiratory movements may also
be altered by local changes in the lungs, air tubes, pleura, respiratory
muscles and nerves, etc.
TECHNICAL NOTES ON METHODS OF STUDYING THE RESPIRATORY
MOVEMENTS
The present chapter will deal mainly with modifications in the respira-
tory movements, their rate and amplitude, etc. These may be abserved
and counted directly, or they may be recorded by registering the t \cursion
of the chest walls or diaphragm or the passage of air from the lungs or pleura.
Respiratory Tracings.— These may be taken on a separate drum, moving
at the same speed as that used for recording tin- blood-pressure. The
lexer-. CtC., are adjusted SO that the excursion <>! the normal respiration
i height of i to i inch on the drum. The Inning should be marked
to show whether inspiration ( orre-pond- to the upstroke or downstrokc.
il method- \\ill he described; none is universally -at isfactory.
i. Trachea-tambour Method.— This is the simplest method, commonly
used in an< sthrti/rd animals. The trachea! eannula is connected by
wide tubing with a laru'e T piec c-. The second limb of the T bears a short
piece of tubing which can be narrowed by a screw-clamp. The third limb
240
A LABORATORY GUIDE IN PHARMACOLOGY
is connected with the recording tambour. The screw-clamp is adjusted
so that the lever-point makes the desired excursion. In place of the screw-
clamp a hole may be cut in the tubing, which can be partly occluded by a
piece of glass-rod inserted through the free end (Fig. 46). If the anesthetic
Fig. 46. — Trachea-tambour method.
is to be given, the open end of the T tube is inserted into the mouth of the
anesthetic bottle (not immersed).
This method has the advantage of simplicity and is not disturbed by
movements of the animal. It suffices to register the rate and changes in
l/ent
Fig. 47. — Organ-key bellows recorder. Actual size.
the depth of the respiration; but, because of the escape of air, it becomes in-
accurate if the respirations are slow or prolonged.
Recording Tambours. — The cheapest form consists of a home-made organ-key bellows
(Fig. 47), the sides of very thin leather or gold-beater's skin. A 3- or 4-cm. Marey's tam-
bour answers well. The 3-cm. Brodie bellows (made by C. F
Palmer, 6 Upper Tulso Hill, London, N. W.) is the most deli-
cate. All bear a straw and writing point about 6 inches long.
2. Trachea-bottie-tambour Method. — This avoids the in-
accuracy of the preceding method by interposing a large
closed bottle into which the animal breathes while the
record is taken, but it introduces the complication of more
or less asphyxia.
The arrangement is explained by Fig. 48. The bottle
should be as large as possible (a 5 -gallon glycerin can or large
jug answers). The connection between the trachea and
bottle should be as short and wide as possible. The vent is.
closed whenever tracings are taken, and opened between the
tracings. The greatest care must be used to avoid asphyxia.
It is advisable to disconnect the bottle occasionally and blow
air through it with bellows.
~Tra.ch.ea-
f **
Fig. 48.— Respiration bottle. 3. Mask-tambour Method. — The air may also
be taken from a mask fitting air-tight over the
mouth and nose and provided with a T piece, as in the first method.
A Henderson "tennis-ball cardiometer" is very satisfactory.
4. Nasal-tambour Method (Unanesthetized Rabbit). — This corresponds to the first
method, except that a cannula, expanded into an olive-shaped bulb at the tip, is inserted
into a nostril of the animal, and fixed with adhesive plaster, if necessary, and connected
with the tambour (Wolff, 1913, Arch. exp. Path., 74, 299).
CHAP. XLI
RESPIRATION (AND BLOOD-PRESSURE)
241
5. Double Tambour Method Stethograph . — This can also be used for non-anes-
thetized animals. A large tambour or other elastic reservoir is tied firmly to the chest
or abdomen. Its interior is connected to a recording tambour, with the interposition of
a T piece, by means of which the tambours can be moderately distended.
The receiving tambour may be given various forms. An efficient instrument may be
made by cutting off the top of a pound ether tin a centimeter below the rim, tying a rubber
membrane over this, and closing the stopper opening with a perforated cork, bearing a
glass tube.
The sleeve of the sphygmomanometer can be wound about the chest and connected
with the recording tambour; or a piece of bicycle tire will answer the purpose.
6. Lever Methods.— In these the motion is transmitted to an ordinary muscle-lever.
may be done (i) by taking a stitch through the skin and tying the string to the lever.
(2) A small incision may be made through the skin and muscle, on the right side, about the
lower edge of the diaphragm; the end of a glass rod or the bowl of a teaspoon is inserted
between the liver and diaphragm and the handle connected with the lever. (3) A knitting
needle may be thrust directly into the liver through the skin (danger of hemorrhage!). (4)
A special lever may be used, bearing a rod which rests on the chest and abdomen. This
does not require anesthesia. It is well adapted to obtaining tracings of the Cheyne-Stokes
respiration in deep anesthesia. The animal must be immobilized in all the lever methods.
Fig. 49.— Diagram of Dreser spirometer.
7. Respiratory Plethysmograph for Entire Rabbit.— This is described by Cu>hn\
Jour. Pharmatol., 4. ^n^; ami a -impler form by Cushny and Lieb, 1915, ibid., 6, 451.
8. Pleural Cannula Method. — In this the air is obtained from a flanged cannula in the
thorad. wall.
9. Spirometer Methods. — These measure the total volume of air breathed
. 49). The expired and in>pired air are separated by the valve. Gas-
meters may be used instead of the spirometer.
TECHNICAL REFERENCES ON RESPIRATION
General.— Robert. Iniox., i, 202, 243; £
Observation and Recording. — Tigerstedt, 2.2, 3; II I. 427, 433.
Bellows Recorder. 07.
Respiration Valves, (iuthrii \ssoc., 57, 887.
Spirometer. Drexr. iSs ,:h .. 26, 253; Arch. ges. Physiol., 1898, 72,
prn-. iS«)o. ibid., 78. 529.
Gas Meters.— Tigerstedt, 1.3, 144; for small quantities, Y. Henderson, Amer. Jour.
Physiol., 25, 385, 1910.
16
242
A LABORATORY GUIDE IN PHARMACOLOGY
Carbon Dioxid Test for Respiratory Excitability. — A. Loewy, 1890, Arch. ges. Physiol.,
47, 601 ; in man, Lindhard, 191 1, Jour. Physiol., 42, 337; Y. Henderson (holding of breath as
index of acidosis), 1914, Jour. Amer. Med. Assoc., 63, 318.
Respiration Experiments on Man. — Y. Henderson, 1914, Jour. Amer. Med. Assoc., 62,
1133; Higgins and Means, 1915, Jour. Pharmacol., 7, i.
Respiratory Metabolism. — Tigerstedt, 1.3, 71; Abderhalden, 3, 1143; Man, ibid., 7,
452; 8, 529; Alveolar Air, Comparison of methods, Boothby and Peabody, 1914, Arch. Int.
Med., 13, 497; Micros pi rometer (small organisms), Thumberg, 1905, Skand. Arch. Physiol.,
17, 74-
Alveolar Ventilation and CO2 Tension. — Man, Higgins and Means, 1915, Jour. Phar-
macol., 7, i; Animals, Macht, 1915, ibid., 7, 339.
TECHNICAL NOTES ON METHODS OF RECORDING THE ARTERIAL
BLOOD-PRESSURE
The usual methods of recording the blood-pressure of anesthetized
animals consist in connecting the carotid (sometimes femoral) artery with
a manometer which writes on a revolving cylinder (kymograph). The
general arrangement is shown in Fig. 50.
Fig. 50. — Arrangement for taking a blood -pressure tracing (Stewart): m, Manometer; kg,
mercury;/, float armed with writing-point; a, thread attached to a wire projecting from the drum
and supporting a small weight; the thread keeps the writing-point in contact with the smoked paper
on the drum; b is a strong rubber tube connecting the manometer with the artery; c, a pinch-cock on
the rubber tube, which is taken off when a tracing is to be obtained.
Taking an Ordinary Blood-pressure Tracing. — The manometer (Fig. 51),
containing clean mercury, is clamped to the table. A drop of very thin oil
is placed on the float. The arterial limb of the manometer bears a T-tube
CHAP. XLI RESPIRATION (AND BLOOD-PRESSURE) 243
(not shown in the figure). The horizontal limb is attached to a rubber
tube (which will connect with the artery). A screw-clamp and a strong
pinch-cock are placed on this tube. The vertical limb of the T is connected
with a bulb, placed 4 feet above the table, and filled with half-saturated
magnesium sulphate solution. The stop-cock between the manometer and the
magnesium must always be kept closed when the artery is open, else the solu-
tion will reach the heart and speedily kill the animal.
A drum is smoked and adjusted to the manometer with a guide-thread.
The opening of the arterial tube is raised (on a tumbler) to the position
which it would occupy in the animal. The pinch-cock is removed and the
screw-cock is opened. The magnesium cock is now opened, filling the con-
nections air-free. It is then shut off. This gives the zero pressure in the
manometer. The drum is adjusted so that the writing-point of the man-
ometer is about an inch from the bottom. A signal magnet is adjusted at
the same point. This traces the zero pressure abscissa line.
The artery tube is now clamped, and the magnesium cock opened until
the pressure has risen. The magnesium cock is then closed. The drum is
adjusted so as to move about 2 cm. per minute, and a minute's revolution
is marked off on the abscissa. This serves as a measure of the time for the
whole tracing. (This is not necessary if a time signal is used.) If a respiratory
or other tracing is also to be taken in the same drum, the writing-point is
adjusted on a vertical line from the manometer-point, about i\ inches from
the top of the drum.
The artery cannula may now be filled with magnesium and connected
with the artery tube (making sure that the magnesium cock is closed)
and the tracing started. The screw-clamp on the artery tube is tightened
until the excursions are of moderate degree (3 to 10 mm.). This gives a
more accurate record of the mean pressure, and also prevents the excessive
flow of magnesium into the artery. Injections, etc., are marked with the
signal.
A normal tracing should always be taken before the drug is injected.
Tracings should also be taken during the injection and whenever any
interesting phenomenon occurs. It may be advisable to stop the drum
between these periods, especially if a fast speed is used. This is not often
necessary in using the slow gear and the 10 X 2.2 cm. vane of the Harvard
kymograph, the most generally useful for pharmacologic work. Only a
single round of tracings should be taken on each paper. (It is sometimes
desirable to take both a slow and a fast tracing at the same time, joining
two manometers to the same carotid by a T piece and using two kymographs;
this is especially instructive with digitalis and aconite. The slow tracing
is made continuous, while the fast tracing is only taken at intervals.)
If clotting occurs, i. c., if the manometer ceases to pulsate, the artery
is clamped, the cannula detached and cleaned with a feather, and the
artery tube is llu-lu-d with magnesium.
The actual blood-pressure may be read from the tracing, being t
the vertical distance between the tracing and the abscissa.
TECHNICAL REFERENCES ON GENERAL TECHNIC OF BLOOD-PRESSURE EXPERIMENTS
Abdcrh;il'i . 845; 2, 15^; K««l>rrt. Int..\.. i, 225.
Comparative Vasomotor Reactions in Di/erent Arteries, Gunning, 1916, Ann |..ur.
I.. 41. i.
resents the excursions in but one limb of the manometer, the men ury
in the other limb is. of course, changed by the same amount. The pressure corresponds to the
difference between the two limbs, i. - t hat in one liml ..
244
A LABORATORY GUIDE IN PHARMACOLOGY
MANOMETERS: MERCURY MANOMETER
This consists of a glass tube, bent as shown in Fig. 51. No. 9 tubing is used for dogs,
No. 7 for rabbits. The straight limb is about ic inches high. It may be surmounted by a
T-tube for connection with the magnesium. The tube is mounted on a small board.
A cleat may be screwed to the back of this board, about its middle, project ing an inch on
one side. This is clamped to the table. It should
be leveled so that the vertical tube is plumb.
The board also bears a millimeter scale, with
arbitrary zero point. The manometer is filled
about one-half with mercury. The bent limb is
filled with 25 per cent, magnesium sulphate solu-
tion, and connected with a stiff rubber tube long
enough to reach to the carotid cannula. This
tube is closed with a pinch-cock (or a lead tube
and metal stop-cock may be substituted, but
with little advantage). The connecting tube is
also filled with magnesium solution by means of
a long-pointed pipet. The pressure in the man-
ometer is now raised to about the blood-pressure
of the animal (say 120 mm.). This may be ac-
complished simply by forcibly blowing into the
rubber tube, clamping near the manometer, and
again filling the tube, or the tube may be con-
nected by a T piece with a perfusion bottle filled
with a magnesium solution and raised to the de-
sired level.
For recording the excursions of the manom-
eter the straight limb bears a float, /. This
consists of a little cylinder of hard rubber, of the
shape and size shown in the figure. It should fit
snugly but rather loosely in the tube. It bears a
knitting needle, well centered. This again passes
through a hard rubber cap, c. At the upper ex-
tremity this needle carries a small flat piece of
cork, to which the writing style is attached. This
may be of parchment paper, celluloid, a needle,
or a quill pen. The writing-point should be
bent toward the drum. A few drops of engine
oil should be placed in the tube of the manom-
eter. The mercury must not mount above the
float. The writing-point is held against the drum
by a guide, consisting of a silk thread, suspended
from a wire, and loaded with a ic-gram weight.
The mean blood-pressure equals the differ-
ence between the readings taken at the highest
point reached by the mercury in each limb of
the manometer. It may also be obtained from
the tracing by doubling the distance between the
line of zero pressure and the tracing.
This figure for the mean-pressure is only cor-
rect if the excursions are small or if the systolic
anc1 diastolic variations are of equal duration.
If they are not, the excursions may be reduced
by a screw-clamp on the rubber tube; or the
mean-pressure can be calculated from the trac-
ing. A series of vertical lines are drawn from
the abscissa to the tracing, at equal intervals.
The mean length of these equals one-half the
Fig. 51. — Mercury manometer, one-
quarter actual size; /', section of float,
actual size (Brown).
mean-pressure. This calculation is scarcely necessary in most cases — a little judgment
will enable one to draw the line of mean-pressure approximately without their aid.
The excursions of the manometer with each heart-beat correspond to the pulse-press-
(The excursions of one limb, as seen on the tracing, must be multiplied by 2.)
ure.
The mercury manometer gives only a rough indication of this, the results being viti-
ated by the inertia of the mercury. It also gives a very imperfect picture of the details
of the individual pulse- waves. An elastic manometer (e. g., Huerthle's) is necessary for their
CHAP. XLI RESPIRATION (AND BLOOD-PRESSURE) 245
accurate study. The mercury manometer is especially useful on account of its simplicity
and for obtaining the mean pressure. Very good results are obtained by taking simul-
taneous tracings with both manometers, connecting the mercury with the carotid, and
Huerthle's with the femoral.
Technical References on Manometers. — Abderhalden, 5, 130; Tigerstedt, 2.4, i.
Mercury, Guthrie, Jour. Amer. Med. Asspc., Nov., 14, 1903.
Optical, Wiggers, 1914, Amer. Jo.ur. Physiol., 33, 384; 1915, Jour. Amer. Med. Assoc.,
64, 1305.
Principles of Registration. — Tigerstedt, 1.4, 51.
Purification of Mercury. — Abderhalden, 3, 560, 563.
Signal Magnet. — This is useful for marking the time of injections, stimulations, etc.
The Harvard instrument is efficient. The electromagnet is connected with a battery
(which may be placed under the table), with the interposition of a key, which is closed
whenever a mark is to be made on the drum. It may be kept closed during the duration
of the injection. The writing-point of the signal must be exactly on a vertical line with the
writing-point of the manometer.
Injection Signal. — A simple device for recording automatically the beginning and dura-
tion of injections is described by Chase and Schlomovitz, 1915, Jour. Pharmacol., 6, 561.
Interpretation of Membrane Manometer Curves. — Pilcher, 1915, Amer. Jour. Physiol.,
38, 209.
ANTICOAGULANT SOLUTIONS
Magnesium Sulphate, half-saturated (25 per cent, of crystals) is the most
satisfactory solution for dogs and rabbits. It does not answer quite as well
for cats, or where large pressure changes are anticipated. Care must be
taken, however, that it does not enter the heart, for it causes prompt
paralysis of this organ. The danger of this accident is not great unless too
high a preliminary pressure has been produced in the manometer. The
effects pass off very quickly unless the heart is stopped completely. Should
this occur, it is often possible to resuscitate the animal by artificial respira-
tion, injection of normal salt solution, and cardiac massage. (Magnesium
sulphate must never be used to fill the connection with the injection buret).
Devices for Lessening the Entrance of the Anticoagulant Solution into the Circula-
tion.— A bulb of about is-c.c. capacity, shaped as in Fig. 52, may be inserted horizontally,
next to the arterial cannula.
Fig. 52. — Magnesium bulb.
Other devices are described by Brooks and Luckhardt, 1915, Amer. Jour. Physiol.,
36, 104.
Other Anticoagulant Solutions.— Carbonate-bicarbonate Solution. — Sodium bicarbon-
ate, 4<> ^m.; Sod. ( arbon;i'< .vater, <|. s.. i liter.
Carbonate Solution. — Half -atur.ited; <|uitr toxic.
Sodiu '
Sodium Sulphate— Half saturated. The Sodium Citrate and Sodium Sulphate are
less dangerous, also less etn. i< nt.
iay be used in tin- i annula. as wrll as in the entire animal.
necessary to render the blood of an animal non-coagulable ; for instance,
• :lo\v from veins, or for j»ra< ti^ii ion.
The best method consists in the intravenous injeitimi ni /. For each kilo
of bo< 'he heads of three lee. lies are rubbed with sand and <• . .1 . of 0.9 per irnt.
•; This causes apparently no change in the ( ir. ul.it inn.
ment extracts is described in Abderhalden. 2, 900;
Tigerstedt, 2.4,325; Abel, Jour 1'harmacol., 5,
270. Merck's Extr. Sangisuga sic. comes
246 A LABORATORY GUIDE IN PHARMACOLOGY
in tubes of o.i gm., corresponding to three heads, and sufficient for i kg. of blood. Hirudin
requires i mg. for 5 c.c. of blood.
The same object may be accomplished by the Lewaschew-Pick method of defibrina-
tion. About 20 c.c. of blood per kg. of animal are drawn from an artery into a porcelain
capsule, defibrinated by beating with a glass rod, strained, warmed, and reinjected into a
vein. This is repeated every- half-hour until the blood yields no coagulum. Six or seven
defibrinations are needed for this end. Peptone is tess certain and causes a considerable
fall of blood-pressure; 0.3 to 0.6 gm. of Witte's peptone per kilo are injected intravenously
(as 5 per cent, solution).
TECHNICAL REFERENCES ON BLOOD-PRESSURE IN NON-ANESTHETIZED ANIMALS
Brooks, 1910, Jour. Amer. Med. Assoc., 55, 372; Heart, 2, 5; 1915, Amer. Jour. Physiol.,
36, 104; Van Leersum, 1911, Arch. ges. Physiol., 142, 377; Trendelenburg, 1913, Zs. exp.
Med., 2, i; Robert, Intox., i, 205.
TECHNICAL NOTES ON ORDINARY OPERATIVE ANESTHESIA
Operations Are to Be Made Only Under Complete Surgical Anesthesia. —
The method of anesthesia depends to some extent on the animal (see also
Chapter XLII).
The anesthetic may be administered either by inhalation or by injection.
Inhalation anesthesia is best adapted to relatively short operations; in-
jections are preferred when the conditions must be kept constant for some
time. The combination of both methods is often advantageous.
ANESTHETICS ADAPTED TO DOGS
Morphin-ether Anesthesia. — 10 to 20 mg. of Morphin per kg. (hydro-
chlorid or sulphate, J to \ c.c. per kg. of 4 per cent, solution) is injected
hypodermically (before the laboratory time) and followed in half an hour
or an hour by the inhalation of ether.
Fig. 53. — Ether cone, about one-half actual size.
If the larger dose of morphin has been used, the ether may usually be
withdrawn when the operation is completed, the morphin sufficing to keep
the animal narcotized, except when especially painful procedures are em-
ployed, when it can be again reinforced with ether. Young dogs should
receive relatively less morphin.
Ether Cone for Dogs. — This consists of a conical tin (Fig. 53). The
interior of the bottom, which is open except for the cross-pieces, is lined
with a small handful of cotton. Two copper wires are fastened behind
the ears of the animal to hold the cone in place.
Administration of Ether to Dogs. — The operator kneels over the animal,
holding it firmly behind the ears. A tablespoon of ether is poured into the
CHAP. XLI
RESPIRATION (AND BLOOD-PRESSURE)
247
cone, and this is fastened on the animal and tightened with a towel. More
ether is added as needed, and if the animal is not anesthetized in a reasonable
time, the holes are occluded with the hand. Complete muscular relaxation
is the best sign of adequate anesthesia. The anesthetist must keep his
attention constantly on the animal, and regulate the anesthetic and supply
air or artificial respiration as needed. During long operations the animal
must be kept warm with towels, etc.
u
Fig. 54. — WoulfFs bottle for giving anesthetic (also arranged for respiratory tracing).
When the trachea has been opened, the tracheal cannula is connected by
a tube with a WoulfT bottle (250 c.c.) containing cotton moistened with
ether. The concentration of the vapor may be varied by the distance of
the tube from the ether, or by limiting the intake of air.
Fig. 54 shows this arrangement adapted to respiratory tracings. D. E.
Jackson, 1912, Jour. Amer. Med. Assoc., 58, 475, describes a special ether
valve.
The ether may also be given by insufflation (Fig. 55; see also Chapter
55-— Ether by insufflation
I-'.tluT may, of course, also be used without morphin, but is much less
u tnrv and much more dangerous.
Other inhalation anesthetics may be substitute*! for the ether, bu( are leas satis i.
•M-ral use.
..r an I ( / \lixtnrr ..-.,ual parts of Alcohol, Chloroform, and I
• tin- 1>I<«>.I prc^un- and arc more dangemu-.
'•i.irt operation- It may be given without morphin by
spray < ."l in the bottom of a tumbler. whi< h i* then invrrtoi
thr mouth and \\rap; : t< r results are secured by
through a special mask. Kitrous Oxid is also useful for very short operations.
248 A LABORATORY GUIDE IN PHARMACOLOGY
Grehant Anesthesia. — This is one of the best injection methods, giving
a lasting anesthesia. The animal is given a hypodermic injection of o.oi
gm. per kg. of morphin (J c.c. per kg. of 4 per cent.), followed in half an
hour by 6 to 10 c.c. per kg. (according to age) of the mixture, diluted with
water to make a total of 200 c.c., administered by the stomach-tube. The
anesthesia is complete in five to fifteen minutes and lasts for eight to
fourteen hours. The mixture consists of chloroform, 50 c.c. ; alcohol and
water, each 500 c.c.
Morphin-chloretone Anesthesia. — This is similar to the Grehant, but cannot be used
if the animal is to recover from the operation. The morphin is injected as above. After
one to three hours 0.2 gm. per kg. of chloretone, dissolved in a small quantity of alcohol,
is injected through a stomach-tube.
Chloral. — This is but little used. The dose is 0.25 to 0.3 gm. per kg. by stomach; o.i
to 0.15 gm. per kg. by vein.
ANESTHETICS ADAPTED TO CATS
Morphin-atropin-urethane. — This has been the most satisfactory
cat anesthetic in this laboratory. The atropin is intended to prevent
reflex-vagus stoppage. It cannot be used if the vagus is to be studied.
The mixture contains morphin sulphate, i gm.; atropin sulphate, 20 mg.;
ethyl carbamate, 20 gm. ; water, q. s. 100 c.c. Of this solution, 3 c.c. per kg.
are administered by stomach or, preferably, by rectum a full half-h6ur
before the operation. The animals appear conscious, but may be tied and
operated without resistance or other signs of pain; but it is advisable to
give a little ether during the operation.
With a little experience cats can be handled without any danger, but
it is safer to wear gloves.
Morphin-chloretone. — Edmunds and Gushing place the animal in a box 35 cm. long,
1 8 cm. wide, and 18 cm. deep. The box is furnished with a sliding lid. A V-shaped cut
is made in the end of the lid and in the corresponding end of the box, so that the animal
may be securely clamped in this opening, allowing the head to protrude. The lid
is fixed with a nail; 40 to 60 mg. of morphin are injected with a hypodermic syringe into
the skin of the neck. This is followed by 0.3 gm. per kg. of chloretone dissolved in alcohol,
administered by a stomach -tube.
The chloretone (same dose in oil) may also be injected into the peritoneum.
Ether or Chloroform-ether (Equal Parts). — The animal is placed in a tight box or
bell- jar, into which are dropped sponges saturated with the anesthetic until the required
degree of anesthesia is procured.
ANESTHETICS ADAPTED TO RABBITS
Morphin-urethane. — Morphin, 5 mg. (J c.c. of 4 per cent.) per kg.,
hypodermically, with ethylcarbamate, 0.75 gm. per kg. by stomach or 0.5
gm. per kg. by rectum.
Or urethane alone, i gm. per kg. by stomach or 0.75 by rectum; or Chloral, 0.6 gm. per
kg. by stomach, 0.3 gm. per kg. per rectum; or Paraldehyd, i gm. per kg. by stomach; or
Chloretone, 16 c.c. of saturated watery solution per kg. (often fatal) may be substituted.
The analgesics may be supplemented after fifteen or twenty minutes
by light and careful etherization, but this is rarely necessary. Rabbits
bear chloroform very badly.
ANESTHETICS ADAPTED TO MONKEYS
Morphin, 30 mg. for small, 60 mg. for large animals, followed by ether.
CHAP. XLI RESPIRATION (AND BLOOD-PRESSURE) 249
ANESTHETICS ADAPTED TO SMALL ANIMALS
Mice, guinea-pigs, rats, etc.: Ether.
ANESTHETICS ADAPTED TO FOWL
Paraldehyd, 2 c.c. per kg. by rectum (Edmunds and Roth, 1908); or Atropin, 0.3 mg.
hypodermic-ally, followed immediately by ether (Pearl and Surface, 1009, Jour. Amer.
Med. Assoc., 52, 382).
DECEREBRATION
This was described in Chapter XXXIV, page 160.
INTRACEREBRAL MAGNESIUM CHLORID
Henderson, Jour. Pharmacol., i, 109.
SPINAL ANESTHESIA
Tigerstedt, 3.4, 8.
OPERATIVE TECHNIC
Animal Boards. — For convenience in operating the anesthetized animals should be
tied to a board. A number of complicated holders are in use, but the one illustrated in
lyg. 56 is cheap and answers almost every purpose. It should slope gently toward the
feet.
A number of sizes should be on hand for different sized animals (i by 4 feet for dogs;
8 by 30 inches for rabbits). The cross-piece is made of wire ,36 inch in diameter. It is
Fig. 56.— Dog board.
pu-hed back of the canine teeth. A 2-foot piece of stout twine1 is passed under the mvk,
behind the ears, the ends are brought forward, wound tightly around the wire, and tied
about the mouth. This holds the head very securely. In operating on the mvk the front
;<>uld be tied toward the abdomen; in operating on the chest, they are secured toward
the head.
In prolonged operation*, it may be advisable to heat the animal to prevent shock.
Technical References. — A m'nuil Holders. — Tigerstedt, 2.4, 1 1 .
^Brodie Operating Table.— Title -nirer, 116.
Operative Dissections.— The hair of the animal should be well clipped
over the field of operation. Scissors (6-inch) , curved on the flat, are efficien t .
The >nialler cut hairs are removed by a wet sponge. Tin- wound should
be kept as free from blood as possible. This frequently determine- the
•ilure of a delicate operation. Incisions should be made, it"
:!>lc, in tin- median line; the muscles and fasciae should be separated by
blunt desertion. Bleeding vessels are secured by hemnstats and tied.
Blood i- removed by spon.uin.u with -mall pieces of absorbent tot ton.
Practically no blood should be lost in operating on the neck, ijroin. or
abdomen. The wound may l>e spread by tcnaeuli, etc., or by \\ eights
iied with a cord to hooks.
.1 hemp No. 3 for dogs; dauntless flax No. 24 for rabbits.
250 A LABORATORY GUIDE IN PHARMACOLOGY
Operations on the Neck. — The forelegs are tied toward the tail. The
structures are most conveniently reached by a median incision, from the
lower end of the larynx to near the sternum. The tissues should be divided
by layers, keeping to the median line, until the trachea is reached. This
may be lifted with the fingers and cleaned with the forceps. Tracheotomy
is the first step in pharmacologic experiments, as it facilitates anesthesia
and artificial respiration. By feeling outward from the trachea, at the bot-
tom of the wound, the carotid artery may be felt pulsating. It is lifted to
the surface with the fingers, or by turning the edge of the wound outward.
The vagus nerve in the dog lies in the same sheath as the artery, and must
be carefully and gently separated from it. It should never be included
in the arterial ligature. In the dog the vagus trunk includes the sym-
pathetic and depressor fibers. These run separately in the rabbit, but
all in the immediate neighborhood of the artery; they may be recognized
by their size, the vagus being the largest, the depressor the smallest. (Illus-
tration in Heinz, I, p. 730.)
Stimulation and Division of Nerves. — Nerves must always be manipu-
lated gently. If it is desired to stimulate or divide the vagus or any other
line,
Leg Abdomen
Fig. 57. — Diagram of dissection of femoral vessels of dog (Brown).
nerve later in the experiment, a ligature may be passed under it and the
ends knotted. The nerve can thus be easily found and lifted from the
wound. In other cases it may be desirable to divide the nerve, securing
each end with a ligature. Nerves should always be protected from drying,
leaving them in the wound, if possible. In electric stimulation, good
contact of the electrodes should be secured. Stimulation of adjacent
structures may be prevented by slipping a strip of rubber-dam under
the electrodes, or by the use of "shielded" electrodes.
For the dissection of the Accelerator Nerves, see Practical Physiology, Beddard, etc.;
or Heinz, I, p. 726. A preliminary dissection is indispensable.
The external jugular vein is exposed by blunt dissection between the
skin and muscle. If offers no difficulty. It may also be reached directly
by a skin incision made about the middle of the neck, in a line drawn from
the angle of the jaw to the manubrium.
The thoracic duct may also be isolated in the base of the neck. It terminates in the left
subclavian vein. A practice dissection is necessary.
CHAP. XLI RESPIRATION (AND BLOOD-PRESSURE) 251
Exposing the Femoral Vessels. — These may be felt pulsating just below
Poupart's ligament on the outer edge of the stiff adductor longus muscle.
The artery lies partly behind and external to the vein (Fig. 57). The
cannulae should be introduced as high as possible. As the vessels give off
branches in this region, the dissection must be made carefully.
The Sciatic Nerve. — To expose this the hind leg is held up, and an
incision is made through the skin in the median ridge of the posterior sur-
face of the thigh. The muscles are separated with the fingers, keeping a
little outward from the middle line. The nerve is felt at the bottom of the
wound as a hard cord. The animal should be in deep anesthesia when the
nerve is handled.
Control of Hemorrhage. — Visible blood-vessels are clamped or tied;
capillary hemorrhage is arrested by packing with cotton or Pangawahr
Djambi; by pressure with the cut surface of a piece of muscle (V. Horsley,
Brit. Med. Jour., July 4, 1914); or by actual cautery.
References. — Tigerstedt, i.i, 40. The preparation of henwstatic tissue extract is de-
scribed by Hess, 1915, Soc. Exp. Biol. Med., 12, 117; Hirschfelder, 1915, Berl. Klin.
Woch., 976.
Technical References on Operative Technic. — Tigerstedt, i.i, i; 2.4, 322.
EXERCISE I.— (DEMONSTRATION) MORPHIN, ETC., ON VOLUME OF
EXPIRED AIR
(REPORTER I, D)
Arrange a Dreser spirometer (see Fig. 49). If anesthesia is permissible
a tracheal cannula should be used. When observations are to be made
without anesthesia a mask (cardiometer bulb) is applied. The rabbit is
tied on a board or confined snugly in a box, with only the head protruding.
Experiment i. Effect of Morphin on Normal Rabbit. — Connect ap-
paratus with nostrils. When animal has become accustomed to the ap-
paratus, close the side tube, starting the collection of the expired air and
the tracing. Collect the air for one minute; then disconnect from spi-
rometer, but continue respiratory tracing.
Inject hypodermically a therapeutic dose of morphin, 0.5 mg. (J c.c.
of i : 1000) per kg. Repeat observations at intervals.
Experiment 2. Morphin in Hyperpneic Rabbit. — Place a normal rabbit
in a box so that it can be heated by hot-water bottles.
normal observations. Heat the box until hyperpnea becomes
pronounced. Take observations. Inject morphin as in Experiment i.
Take observations.
Remove heat, and when temperature has become normal take obser-
vations.
Experiment 3. Toxic Dose of Morphin.— Inject hypodcrmieallv into
the rabbit of Experiment i a toxic dose of morphin, 40 mg. (i c.c. of 4 per
cent and observe results.
Experiment 4. Camphor After Morphin. — Use morphinized rabbit of
Experiment 3, After taking normal observation inject into peritoneum
camphor o.i gm. (0.5 c.c. of 20 per cent, in oil) per kg.
Experiment 5. Caffein After Morphin. — Use morphinized rabbit of
Experiment I, After taking normal ol»rr\ ation inject hypodermically
caffein, 10 mg. (i c.c. of i per eent.) per kg.
252 A LABORATORY GUIDE IN PHARMACOLOGY
QUESTIONS
(a) What are the effects of morphin on normal respiration — rate;
depth; minute volume; single volume?
(b) How are these effects modified in dyspnea?
(c) Under what conditions would morphin be most efficient therapeut-
ically?
(d) Describe the effects of toxic doses of morphin and state how they
differ from therapeutic doses.
(e) Describe the effects of camphor.
(/ ) Describe the effects of caffein.
(g) Would these be suitable antidotes for morphin?
(h) In what pathologic states would they be useful?
EXERCISE H.— (OPTIONAL) BRONCHIAL CHANGES
Dreser's method may be used to study the effect of bronchioconstrictors (pituitary)
and bronchiodilators (epinephrin, lobelin, etc.) on the respiratory volume. (See Chapter
XXXVII.)
EXERCISE HI.— (OPTIONAL) RESPONSE OF RESPIRATORY CENTER TO
C02
(See Loewy, 1890, Arch. ges. Physiol., 47, 601.)
EXERCISE IV.— (GROUP I1) RESPIRATION OF NORMAL RABBIT
(REPORTER II, D)
Use rabbit with mask-tambour-T-piece method (Tech. Note.), tracing
on drum. Observe the effects of the following drugs:
Experiment i. Auditory Reflex. — Ring bell near rabbit.
Experiment 2. Counter-irritation. — Rub some Capsicum-petrolatum on
skin.
Experiment 3. Chloral. — 0.5 gm. (20 c.c. of 2.5 per cent.) per kg., by
stomach-tube, when depression is pronounced.
Experiment 4. Hypodermic Irritation. — Inject water, i c.c. per kg.,
hypodermically.
Experiment 5. Caffein. — Hypodermically, 10 mg. (i c.c. of i per cent.)
per kg.
The respiration increases and the animal may come partly out of the
anesthetic (stimulation of the respiratory and other centers).
QUESTIONS
(a) How does the respiration respond to reflex stimuli?
(b) Describe the respiratory effects of chloral.
(c) Describe the respiratory effects of caffein.
(d) What measure could be used against respiratory depression?
(e) Which of these would act most promptly?
' Distribution of Work for Exercises IV to VI (Groups I to HI) :
Student A — Director.
Student B — Weigh animal, give injections.
Student C— Cleaning.
Student D — Reporter; calculate doses.
Student E — General assistant.
Student F — Respiratory tracing.
CHAP. XLI RESPIRATION (AND BLOOD-PRESSURE) 253
EXERCISE V.— (GROUP H1) RESPIRATION OF NORMAL RABBIT
(REPORTER II, D)
Arrange the experiment as in Exercise IV.
Experiment i. Hypodermic Irritation. — Inject water, i c.c. per kg.,
hvpodermically.
Experiment 2. Hypodermic Alcohol. — Inject 50 per cent. Alcohol,
i c.c. per kg., hypodermkally.
Experiment 3. Strychnin, Therapeutic Dose. — Inject hypodermically
a therapeutic dose of Strychnin, 0.2 mg. (0.2 c.c. of i : 1000) per kg.:
increased respiration (stimulation of respiratory center). Reflexes in-
creased (increased excitability of spinal cord).
Experiment 4. Atropin, Therapeutic Dose. — Inject hypodermically
Atropin, i mg. (i c.c. of i : 1000) per k<j.
QUESTIONS
(a) How does the hypodermic injection of whisky act on respiration?
(b) Describe the respiratory effect of strychnin.
(c) Describe the respiratory effect of atropin.
(d) In what conditions would these be therapeutically useful?
EXERCISE VI.— (GROUP HI2) RESPIRATION OF NORMAL RABBIT
(REPORTER II, D)
Arrange the experiment as in Exercise IV.
Experiment i. Ammonia Reflex. — Blow Ammonia vapor into nostril.
\« >tice respiratory standstill and stoppage of the heart (reflex stimulation
of vagus center by irritation of the trigeminal endings). On removing the
ammonia the respiration is increased (dyspnea) and the heart resumes.
Experiment 2. Morphin; Therapeutic. — Inject hypodermically 0.5 mg.
(J c.c. of i : 1000) per kg.
Experiment 3. Morphin; Toxic. — Inject hypodermically 20 mg. (J c.c.
of 4 per cent.) per kg. Respiration becomes slow and shallow (depression
of rv-piratory centers).
Experiment 4. Nicotin. — Inject hypodermically 0.5 mg. (| c.c. of
i : 1000) PIT k^.
QUESTIONS
(a) Describe the respiratory effects of irritant vapors.
(b) Describe- tin- ropiratory effects of morphin.
(c) DescriU- the respiratory effects of nicotin.
(<T) How could you counteract respiratory depression?
EXERCISE VII.— (GROUP IV3) RESPIRATORY AND BLOOD-PRESSURE
TRACINGS
D
Inject a dog hypodermic ally with a small dose of Morphin, 10 mg.
(} c.c. of i per cent.) p«r k- Aitn half an hour anesthetize with et In i .
1 Sec foot-note, page 252.
sSf< i>age 252.
> Dist : Work for Exercises VII and VIII (Groups IV and V) :
Student D— Din . tor m.l Reporter; calculates doses; takes notes; prepares report.
r itor.
• -uscilation; cleaning.
re traiinK.
Student F— Res crvation and tracings.
254 A LABORATORY GUIDE IN PHARMACOLOGY
Connect trachea for respiratory tracing (Fig. 54); carotid for blood-pres-
sure, and femoral or vein for injections (Tech. Notes). Determine the
effects of the following drugs and procedures:
Experiment i. Lactic Acid. — Intravenous, 2 c.c. of 0.6 per cent. ( = — )
per kg.
Medullary stimulation: increased respiration; slowed heart; moderate
rise of blood-pressure.
Experiment 2. Caffein. — Intravenous, 20 mg. (2 c.c. of i per cent.)
per kg.
Experiment 3. Camphor. — Vein, o.oi gm. (i c.c. of i per cent, in 40 per
cent, alcohol) per kg.
Experiment 4. Reflex Stimulation by Dilation of Anal Sphincter.
Experiment 5. Reflex stimulation by electric stimulation of sciatic
nerve with weak, moderate, and strong currents.
Experiment 6. Strychnin, Therapeutic Dose. — Hypodermic, 0.05 mg.
(0.05 c.c. of i : 1000) per kg.: often no effects. Sometimes slight increase
of respiration. Circulation little changed.
Experiment 7. Strychnin, Toxic Dose. — 0.25 mg. (J c.c. of i : 1000) per
kg., intravenous; repeated every ten minutes till death. (The dose which
is advised is tetanic in normal dogs, but the effect may be diminished by the
anesthesia.) Before the onset of the tetanus the respiration is increased,
but the circulation is little altered. With the sudden onset of the tetanus
the pressure rises abruptly (central vasomotor stimulation) , and then falls,
with the cessation of the spasm, to a point considerably below normal
(central vasomotor depression); the heart is quick during the spasm (in-
hibition of vagus) ; slow and strong after (vagus stimulation) . The respira-
tion is rapid during the tetanus, depressed in the intervals. Note that the
spasms can be brought on by jarring the table or by blowing on the animal.
The spasms become successively weaker and the blood-pressure does not
rise so high (depression of the convulsive and vasomotor centers). The
heart remains rapid (depression of vagus center) but strong. The respira-
tion ceases (paralysis of the center) and the pressure falls. Begin artificial
respiration at once: the animal can be kept alive almost indefinitely. The
heart and respiration remain fairly good. Note that the pressure varies
with the efficiency of the artificial respiration. Let the animal die. Note
the early onset of rigor (due to tetanus) .
QUESTIONS
Describe the effects of:
(a) Lactic acid (acidosis).
(b) Caffein.
(c) Camphor.
(d) Dilation of anal sphincter.
(e) Sciatic stimulation.
(/) Therapeutic doses of strychnin,
(g) Toxic doses of strychnin.
(h) What is the cause of death in strychnin poisoning?
(i) What are the possible causes of the blood-pressure rise during the
strychnin convulsions?
(j) How could these be distinguished?
CHAP. XLI RESPIRATION (AND BLOOD-PRESSURE) 25$
EXERCISE VIE.— (GROUP V1) RESPIRATORY AND BLOOD-PRESSURE
TRACINGS
(REPORTER IV, D)
Arrange the experiment as in Exercise VII, but record the respiration
by a lever connected with the thorax. The trachea is not opened. Deter-
mine the effects of the following drugs and procedures:
Experiment i . Ammonia Inhalation. — Let the animal breathe Ammonia
vapor.
Experiment 2. Ammonium Chlorid. — Vein, 0.15 gm. (15 c.c. of i per
cent.) per kg.: rate and force of respiration increased, blood-pressure rises;
heart variable (stimulation of medullary centers). Respiratory excursions
increased (heightened excitability of vagus center). The effects are quite
short (rapid elimination) and are not produced by oral administration.
Experiment 3. Mild Asphyxia. — Attach a long tube to the trachea so
as to increase the dead space.
Experiment 4. Severe Asphyxia. — Clamp the trachea until the respira-
tion just stops. Revive by artificial respiration. The respiration, especially
the inspiratory efforts, will first be increased (dyspnea, stimulation of the
respiratory center); then it will be lessened, with rare, gasping, powerful
respiratory efforts (depression of center) ; the blood-pressure rises during
the dyspnea (stimulation of vasomotor center) , and will then fall ; the heart
rate is greatly slowed, with typical strong vagus beats (stimulation of vagus
center). During the dyspnea the animal makes convulsive movements and
the pupils dilate (stimulation of the corresponding centers). The pupils con-
tract again when the paralysis occurs.
Experiment 5. Apnea. — Keep up a brisk artificial respiration for a few
minutes. Stop suddenly, and observe that the animal does not breathe for
some time, the circulation being good. This apnea is due to the fact that
there is not enough CO-2 in the blood to stimulate the respiratory center to
its rhythmic activity.
Experiments 6 and 7. Strychnin, Therapeutic and Toxic Doses. — See
Experiments 6 and 7 of Exercise VII.
QUESTIONS
Describe the effects of:
(a) Ammonia inhalation.
(b) Ammonium injection.
(c) Mild asphyxia.
(d) Severe asph v.\ i a .
(e) Apnea.
(/ to k) Strychnin as in Exercise VII.
EXERCISE IX.-(OPTIONAL) PULMONARY EDEMA
Produce pulmonary edema \>y the method^ mrnti<>nr<l l>r!<>\\. <>!>-< rvr the blood-
pressure, the au>< iiltalion < Manors. an«l ihr foaming: Pilorarpi". MiiMarin. Methyl Sol-
i inhalation. Partial • lamninu «»f aorta with ..vert- «• and
: measures are <lc^< ribed by Miller ,md Matthews. .\r« h. Int. Mr<!.. < >. t.. igpOj 1
Pearce, ibid., June, IQOO; H.dlion ami N'epper. ion. h. Hioph.. i ;. 886.
Try i t the following' methods of treatment: \Ym-so lion, oxygen insufflt-
••inephrin, nitrite-. atn>pin. a~pid"-peniiin. KtrOphantUo.
Edema of Perfused Lung. M \, Arch. gea. Physiol., 158, 509, 527.
' See foot-note, page 253.
256 A LABORATORY GUIDE IN PHARMACOLOGY
CHAPTER XLII
ADMINISTRATION OF ANESTHETICS ON CIRCULATION AND
RESPIRATION
(REPORTERS: C MEMBERS OF EACH GROUP)
Introduction. — Exercises I and II purpose to compare the rapidity and
duration of various anesthetics, simple and "combined" with morphin and
scopolamin.
Exercises III to VI illustrate the effects of the inhalation anesthetics on
respiration and circulation, and the modifications by asphyxia, reflexes,
etc., which are liable to arise during anesthesia. The experiments under
each exercise are so numerous that it will not always be possible to carry
them through; the groups will proceed as far as the condition of the animal
and other circumstances permit.
Exercise VII illustrates the treatment of the accidents arising in anes-
thesia.
TECHNICAL NOTES
Exposure of Kidney and Other Abdominal Organs. — To avoid shock
the exposure of the abdominal organs should be limited as much as possible
both as to area and time. The organs should be kept warm by packing
with cotton, and a can filled with hot water should be kept near the animals.
The abdominal incision is made by preference along the linea alba, toward
the symphysis pub is. This permits the exposure of loops of intestine, of
the spleen, of the bladder and uterus, and of the ureters where they end in
the bladder.
The ureters may be seen posteriorly by lifting out the bladder. (Not to
be confused with the spermatic cord !)
(In male animals the incision through the skin is carried just to one
side of the penis, the superficial veins are ligated and divided, and the dis-
section is carried along the fascia until the linea alba is reached.)
To expose the kidneys, from the median incision, it is necessary to
carry this to near the sternum and to make a second, transverse incision
along the lower border of the ribs. They may also be reached from the
back by an incision about 2 inches from the spine, from the lower border
of the rib obliquely downward. If the incision is made to follow the direc-
tion of the muscle, there need be very little bleeding. The spleen and in-
testine may be reached through the same incision.
Oncometry.— See Chapter XXXV.
Artificial Respiration. — This may be maintained in intact animals
by alternate rhythmic pressure on the chest and abdomen. Very little
force should be used. In operated animals the artificial respiration is
maintained through some mechanical apparatus connected with the tracheal
cannula.
The simplest device consists in a large bellows (15 by 22 inches, exclusive of the handles).
This may be arranged for foot power by fastening a spiral upholsterer's "lounge spring No.
2" between the handles. The spout is closed with a cork. An inch hole is bored in the
top. This bears a perforated cork, from which a tube leads to the tracheal cannula. A
T piece is inserted in the course of this tube, the free limb of the T being closed when the
air is driven into the lungs, and opened when it is expelled. This may be done with the
finger, but it is better to employ some automatic device. The T piece may be placed
directly in the cork of the bellows. The free limb is connected with a rubber tube which is
CHAP. XLII
ADMINISTRATION OF ANESTHETICS
257
tied to the handle in such a fashion that it is stepped on and closed when the bellows is
compressed (Fig. 58). (The spring may also be placed inside of the bellows.)
K. K. Hall has perfected a simple valve for this purpose (Fig. 59). It consists of a
metal T piece, with a steel plunger, well fitted and oiled, which is driven up by the bellows
and falls back in expiration. The excursions are controlled by short pieces of rubber
tubing inserted in the brass.
Trachea.
Fig. 58. — Bellows for artificial respiration.
In an emergency the operator can inflate the lungs by blowing into the
tracheal tube.
Artificial respiration should be performed at about the rate of the
operator's own breathing.
Bellows
Fig. SQ.— Hall's rcs|.r .. Actual nice.
The anesthetic may be continued during the artii'uial re-piration by
the air through the Woulff bottle. >lm\vn in Fig. 54 (taking care
to have the level of the anesthetic so low that it cannot ted into
the tube).
17
258 A LABORATORY GUIDE IN PHARMACOLOGY
Technical References on Artificial Respiration. — Tigerstedt, i.i, 42; Meltzer, 1913,
Jour. Amer. Med. Assoc., 60, 1407.
Pumps. — Pittenger, 106; Gates, 1915, Jour. Pharmacol., 6, 611; Hanzlik, 1916, Jour.
Lab. Clin. Med.
Interruption of Air-blast. — Gesell and Erlanger, 1914, Amer. Jour. Physiol., 33, Proc.
XXXIII.
Pressure Respiration. — Abderhalden, 6, 537.
INSUFFLATION RESPIRATION
A continuous supply of oxygen or compressed air (or glass-blower's
bellows) is connected through two T-tubes, provided with stop-cocks so
that the air may be passed either through an ether bottle or directly to a
catheter with the tips cut off (6 to 7 mm. diameter for dogs; No. 20 French
scale for animals of 9 kg.; Nos. 17 to 1 8 for 7 to 8 kg.).
A dog is anesthetized with morphin and ether. The trachea is opened
and the catheter pushed down until it meets a resistance (i. e., when it has
entered a bronchus). It is .then retracted about 4 cm., which brings it
just above the bifurcation of the trachea; the air current is started. (The
catheter may also be introduced through the larynx; see Meltzer, 1912,
Zentr. Physiol., 26, 204).
The current should be strong enough to have a pressure of 40 to 60 mm.
and to distend the lungs moderately. The opening in the trachea should be
large enough, and the catheter small enough, so that the air may escape
freely. If the stop-cock to the ether is opened and that to the air closed, the
animal receives "full ether"; if both cocks are left open, it receives "half
ether"; if the ether cock is closed and the air-cock opened, it receives pure
air (see Fig. 55, page 247).
References to papers of Meltzer and Auer, Jour. Exp. Med., 1909, n, 622; Jour.
Amer. Med. Assoc., 1911, 57, 521; Zbl. Physiol., 23, 443; ibid., 1912, 26, 204; Jour. Amer.
Med. Assoc., 1913, 60, 1407; ibid., 1914, 62, 1547.
CARDIAC TRACINGS
Simple but rather imperfect tracings are obtained with the intact
chest by acupuncture; 'or with opened thorax by simple levers; but spe-
cially constructed cardiographs or plethysmographs are needed for reliable
tracings.
Acupuncture. — The most convenient method consists in thrusting a knitting needle
through the left thorax, a little above the apex-beat, directly into the heart. This causes
practically no disturbance in the circulation. Another knitting needle is tied securely to
the long limb of a muscle-lever. The two needles are connected by a string. Raising the
string on the heart needle or lowering it on the lever needle will increase the excursions.
The best results are obtained by adjusting the string in the direction of the movement of
the heart needle.
Operation of Exposing the Heart for Cardiographic Tracings. — Dogs
serve the purposes of the experiment much better than cats. They should
be deeply anesthetized (morphin 20 mg. per kg. followed by ether) ; a tracheal
cannula inserted and the artificial respiration apparatus at hand; a motor-
driven bellows with rapid, small excursions gives excellent results; oxygen
insufflation may be employed.
The sternum is laid bare by a median incision extending from about
the second rib to the ensiform cartilage. Hemorrhage is best controlled
CHAP. XLII ADMINISTRATION OF ANESTHETICS 259
by the cautery, but hemostats may be needed. Artificial respiration is
then started. The pericardium is exposed by sawing through the sternum,
care being taken to follow the median line. The saw causes much less
hemorrhage than the knife. Hemorrhage is again best controlled by the
cautery. The sternal edges are separated by hooks attached to the operat-
ing board.
After carefully checking all hemorrhage the pericardium is opened and
the cardiometer applied.
If the animal is restive, so that the position of the cardiometer is dis-
turbed, curare should be used. A towel or large sponge soaked in warm
water should be kepi over the thoracic opening between observations.
Technical References. — Tigerstedt, 2.4, 327.
Tracings From the Exposed Heart. — The thorax is opened under arti-
ficial respiration, as described. The heart is exposed, the pericardium is
divided, and tracings taken. The opened pericardium may be stitched to
the sides of the chest, forming a little hammock for the heart. It is generally
advisable to curarize the animal.
Cardiomyographs. — A hook may be inserted in the apex of the heart
and connected with an elbow lever. However, this is so easily disturbed
by the respiratory and other movements that it is generally unsatisfactory.
The difficulty is largely overcome by the Cushny or Guthrie myograph.
Cardioplethysmographs. — These are conveniently constructed from soft-
rubber balls, as suggested by Y. Henderson (Amer. Jour. Physiol., 1906,
16, 335) : About a third of the ball is cut away and a septum of rubber-dam
cemented over the opening. An aperture is burned through this to fit
snugly to the auriculoventricular groove (several sizes will be needed for
different animals) . The opposite pole of the ball is pierced by a glass tube,
connected with a large tambour, tracing on a drum. Ordinary tambours
may be used for short tracings, but if the heart volume is liable to undergo
material changes the small amount of air may lead to serious pressure on
the heart. It is therefore better, especially if the total volume changes
are to be observed, to employ larger tambours of 10 to 12 cm. diameter.
(These can be constructed from the tops of ether cans.)
In applying the apparatus to the heart the pericardium is cut open
and the ventricle is slipped into the ball, so that the edges of the rubber-
dam fit about the auriculoventricular groove, excluding the auricles. The
cardiimieter is then connected to the recording tambour. The blood-
pressure is a good index of the "fit" of the cardiometer; when the rubber-
dam tit- tu<> tight ly the blood-pressure falls to a low level, while the auricles
are distended.
TECHNICAL REFERENCES
Acupuncture.- , 2.4, 172.
Cardiographs. -Tigcrstcdt, 2.4, 175; Heinz, i, 846; Stewart, 199; Cushny, 1910,
Cardioplethysmographs. : 47; Hm.lerson, 1906, Amrr. Jmir. 1'
16,325: Henderson and Barrinjrrr. [913, i!>i<l.. «i. .•<>.?; LchndnrlY, 1009 (separate .u
and . Arch. Exp. Path . 61, 418; Johannson and Tigerstedt, Skand. An h.
Physiol., i and 2; Santcsson, 1902, ibid., 12.
TECHNICAL REFERENCES ON ANESTHETIC APPARATUS
\- h. Internet. IM . .'2, 487; B<*>thl>v
and Sandifunl. n. c ;m<l Council's anesthesiomctcr),
Jour. Ph .irmu. •>!., 5, 369; Jackson, 1915, Jour. Lab. Clin. V
260 A LABORATORY GUIDE IN PHARMACOLOGY
EXERCISE I.— (GROUP IV, A1) ONSET AND DURATION OF ANESTHESIA IN
NORMAL RABBIT
Observe the narcosis, respiration, reflexes, and especially the time
relations. The animal should be allowed to recover completely between
the anesthetics.
Experiment i. Chloroform Reflex. — Blow chloroform vapor in nostril
of rabbit: temporary arrest of heart.
Experiment 2. Cocainization of Nose. — Fill nostril with cotton saturated
with 2 per cent, cocain. From time to time remove the cotton and try
reaction to chloroform vapor (or ammonia) until this is abolished.
Experiment 3. Nitrous Oxid Anesthesia. — Let rabbit inhale nitrous
oxid through a funnel. Observe effects. Note time of complete anesthesia.
Observe color of mucosae. Are muscles completely relaxed? Remove the
gas as soon as anesthesia is complete. Observe symptoms and time of
recovery.2
Experiment 4. Chloroform Anesthesia. — Pour about 5 c.c. chloroform
on a towel and let rabbit inhale until anesthetized. Observe as in Experi-
ment 3.
Experiment 5. Ether Anesthesia. — Let rabbit inhale about 5 c.c. of
ether, and observe as in Experiment 3.
Experiment 6. Morphin. — Inject hypodermically 10 mg. (f c.c. of
4 per cent.) per kg. Observe effects during half an hour.
Experiment 7. Ether After Morphin. — Repeat Experiment 5 and com-
pare the results.
QUESTIONS
(a) What is one of the dangers of early chloroform anesthesia?
(b) State several means for preventing this.
(c) Tabulate the relative effects of the various anesthetics as to: Res-
piration ; onset of anesthesia (abolition of pain) ; persistence of reflexes ;
muscular relaxation; cyanosis; duration of complete anesthesia after dis-
continuance of the anesthetic; time to complete recovery.
(d) Which of the anesthetics would be best adapted for short operations?
(e) Which would be most dangerous?
(/) What are the undesirable features of nitrous oxid?
(g) How do the morphin effects differ from the anesthetics?
(h) How does morphin modify the course of the anesthesia?
(i) Would its use be advantageous?
EXERCISE II.— (GROUP IV, B3) ONSET AND DURATION OF ANESTHESIA
IN NORMAL RABBIT
Observations as in Exercise I.
Experiment i. Ethyl Chlorid. — Place some cotton in the bottom of a
conical graduate which fits over the face of the rabbit. Pour about 2 c.c.
of ethyl chlorid on the cotton and apply to rabbit, wrapping the cone with
a towel. Observe effects. Note time of complete anesthesia. Observe
color of mucosae. Are muscles completely relaxed? Remove the cone as
soon as anesthesia is complete. Observe symptoms and time of recovery.
1 Distribution of Work for Exercises I and II, Group IV, A, B :
Students C and F — Director and Reporter; administration.
Students A and D — Reflexes and general symptoms.
Students B and E — Respiration; cleaning.
2 Nitrous Oxid as Animal Anesthetic, Dolley, 1914, Jour. Exp. Med., 19, 372.
3 See foot-note No. i.
CHAP. XLII ADMINISTRATION OF ANESTHETICS 261
Experiment 2. Rectal Ether. — Blow ether vapor into rectum. Observe
as in Experiment i.
Experiment 3. Ether Inhalation. — Administer about 5 c.c. on towel.
Observe as in Experiment i.
Experiment 4. Morphin-scopolamin. — Inject hypodermically, per kg.,
morphin 10 mg. 1 1 c.c. of 4 per cent.) and scopolamin § mg. (§ c.c. of
i : 1000). Observe effects during half an hour.
Experiment 5. Ether After Morphin-scopolamin. — Repeat Experiment
3, and compare the results.
QUESTIONS
As hi Exercise I, questions c to A, substituting morphin-scopolamin
for morphin.
EXERCISE m.— (GROUP I) INHALATION ANESTHESIA TRACINGS
Distribution of Work. — Student C — Director and Reporter; narcosis;
observations as below.
Student F — Chief Operator.
Student A — Assistant Operator; weighs animal; calculates doses; gives
injections.
Student B — Anesthetist; artificial respiration and resuscitation; cleaning.
Student D — Circulation observations as below.
Student E — Respiration observations as below.
Observations. — Student C — Narcosis: Reflexes (corneal); muscular re-
laxation; pupils; temperature.
Student D— Circulation: Pulse, blood-pressure tracing, venosity (color)
of blood. (Set up blood-pressure apparatus, pages 242-246).
Student E — Respiration: Count and tracings (set up apparatus, tracheal
tambour method, page, 239).
Accidents. — If the animal should stop breathing, resuscitate according
to Exercise VII (page 264).
Experiment i. Induction of Ether Anesthesia. — Observe pulse, etc., res-
piration, and temperature of normal dog. Pour 15 c.c. of ether into mask
and administer by inhalation. Observe behavior of animal and time till
complete anesthesia (muscular relaxation) .
Operation. — Tie animal to board. Connect carotid for blood-pressure;
trachea for respiration; and femoral vein for injection. For respiration,
numect T-tube, one limb with tracheal cannula, second limb with anes-
thetic bottle, third limb with tracing tambour.
Experiment 2. Reflexes Under Light Ether Anesthesia. — Diminish the
anesthetic unt il reflexes are fairly active, but without spontaneous struggling.
Aim to maintain this >tage. Stretch anal sphincter with artery forceps.
Experiment 3. Insufficient Aeration. — When animal has been brought
back t«» light anesthesia obstruct air passage by partially clamping tracheal
tube.
Experiment 4. Change from Light Ether Anesthesia to Chloroform. —
tore the air- way and bring hark to light am>tlu>ia. Then change
suddenly to chloroform. (In giving chloroform by a mask about 6 to 12
drops are required per minute.)
(Optional) According to S<li;i(iVr i ml S« li.irlirb. the f.ill . f blood-pressure is pr.-u tit ally
prevented by adding 10 per < ml. • •! ;il< <>h«>l t<« • lil.troforn
Experiment 5. Deep Ether Anesthesia. Change back to light ether.
Concentrate the anesthetic to deep ether anesthesia.
262 A LABORATORY GUIDE IN PHARMACOLOGY
Experiment 6. Reflexes Under Deep Ether Anesthesia. — Maintain a
uniform deep anesthesia. Stretch anal sphincter with forceps.
Experiment 7. Insufficient Aeration.— Obstruct air pressure by partially
clamping t radical tube.
Experiment 8. Change from Deep Ether to Chloroform. — Restore
air- way. When conditions have reached constant, change suddenly to
chloroform.
Experiment 9. Reflexes Under Light Chloroform Anesthesia; and
Experiment 10. Insufficient Aeration. — Analogous to Experiments 2 and 3.
Experiment n. Deep Chloroform Anesthesia; Experiment 12. Reflexes;
and Experiment 13. Insufficient Aeration. — Analogous to Experiments 5, 6,
and 7.
Experiment 14. Intravenous Ether Anesthesia. — Withdraw chloroform.
When reflexes return, inject into vein a saturated solution of ether in N. S.,
begin with J c.c. per kg., and regulate flow so as to maintain an even anes-
thesia.
(Optional) Chloroform may be also used by vein: i c.c. of 0.5 per cent, per kg.; see also
Hewitt, 359.
Experiment 15. Chloroform Poisoning. — Stop the ether till reflexes
return. Then let animal inhale chloroform till respiration stops.
Experiment 16. Resuscitation. — Follow Exercise VII, page 264.
QUESTIONS
(a) Tabulate the phenomena of light and deep ether and chloroform
anesthesia.
(b) What are the chief differences between ether and chloroform?
(c) What is their comparative safety?
(d) How do reflexes (operations, etc.) complicate anesthesia?
(e) How does partial asphyxia complicate anesthesia?
(/) Why is it dangerous to change from ether to chloroform?
(g) Is the change to chloroform safer from light ether anesthesia or from
deep ether anesthesia? Why?
(ti) What is the comparative safety of intravenous and inhalation ether
anesthesia?
(i) What are the phenomena of chloroform poisoning late in anesthesia?
(/) Do the clinical chloroform accidents usually occur in this way?
(k) How may chloroform accidents be treated?
(I) Does the same treatment apply to ether accidents?
EXERCISE IV.— (GROUP II) MORPHIN AND INHALATION ANESTHESIA
This is similar to Exercise III, page 261, except that a morphinized
animal is taken, and the kidney volume is also recorded.
Distribution of Work— Observations and Accidents.— As in Exercise III,
except that Student F also observes the kidney oncometer.
Experiment i , a. Morphin. — Observe pulse, respiration, and temperature
of normal dog. Inject hypodermically morphin, 10 mg. (J c.c. of 4 per cent.)
per kg. Repeat observations in half an hour.
Experiment i, b. Induction of Light Ether Anesthesia. — As in Experi-
ment i of Exercise III.
Operations. — As in Exercise III. Also expose kidney and place in on-
cometer, connected with water-manometer or recording tambour.
Experiments 2 and on. — As in Exercise III.
CHAP. XLII ADMINISTRATION OF ANESTHETICS 263
QUESTIONS
As in Exercise III, page 262.
What differences are introduced by the morphin?
From the comparison of the blood-pressure and oncometer changes
deduce whether the circulatory phenomena are vascular or cardiac.
EXERCISE V.— (GROUP V) MORPfflN-SCOPOLAMIN AND INHALATION
ANESTHESIA
This is similar to Exercise III, page 261, except that the animal receives
morphin and scopolamin, and that the volume of an intestinal loop is recorded.
Distribution of Work — Observations and Accidents. — As in Exercise III,
except that Student F also observes the intestinal oncometer and any
grossly visible vascular changes in the intestines.
Experiment i , a. Morphin-scopolamin. — Observe the pulse, respiration,
and temperature of normal dog. Inject, hypodermically, morphin 10 mg.
(} c.c. of 4 per cent.) per kg. and scopolamin § mg. (f c.c. of i : 1000) per
kg. Repeat observations in half an hour.
Experiment i , b. Induction of Light Ether Anesthesia. — As in Experi-
ment i of Exercise III.
Operations. — As in Exercise III. Also expose intestines. One loop
may be placed in an oncometer.
Experiments 2 and on. — As in Exercise III, page 262.
QUESTIONS
As in Exercise IV (morphin-scopolamin in place of morphin).
EXERCISE VI.— (GROUP HI) INSUFFLATION ANESTHESIA AND ANES-
THETIC ACCIDENTS
This illustrates the dangers during full anesthesia.
Distribution of Work. — Student C — Director and Reporter.
Student F — Chief Operator.
Student A — Assistant Operator; weighs animal; calculates doses; gives
injections.
Student B — Anesthetist ; artificial respiration and resuscitation ; clean-
ing.
Student D — Pulse-rate and blood-pressure tracing.
Student E — Cardiograph tracings.
Experiment i, Morphin. — Observe the pulse and respiration of a normal
dog. Inject, hyp<><lcrmi( ally, morphin 20 mg. (J c.c. of 4 per cent.) per kg.
Repeat observation^ in half an hour.
Experiment 2. Induction of Ether Anesthesia. — Administer ether by
cone till animal is deeply anesthetized.
Operation. — Tie animal on board. Insert cannula? into carotid, trachea,
and femoral vein. Start blood-pressure tracing. Start insulllation (page
258) with half ether. Kxpose heart and adjust cardioplethysmograph as
explained <>n pages 258, 259. Start tracing.
Experiment 3. Curare. Inject into vein ;-S rng. (| c.c. of A per cent.)
per kg.; note blno<l-pre»ure ehanges. If neeessary, repeat injection until
movement > im longer interfere with trac ing.
Experiment 4. Excess of Ether.— Change to "full ether."
> (none would on ur for >everal h«
Experiment 5. Excessive Insufflation Pressure.— O!>Mru< t tin mmlow
fr»m the trachea. Watch the blood-pressu lly. so that the effect
264 A LABORATORY GUIDE IN PHARMACOLOGY
does not become too severe. Remove obstructions and let conditions
return to normal.
Experiment 6. Asphyxia. — Interrupt the air current. Again watch
blood -pressure carefully and resume the respiration before it is too late.
Let conditions return to normal.
Experiment 7. Chloroform. — Substitute chloroform for ether, with both
tubes open ("half chloroform").
Experiment 8. Excess of Chloroform. — Change to "full chloroform. "
Before conditions become too dangerous return to "half chloroform."
Experiment 9. Asphyxia. — Repeat Experiment 6.
Experiment 10. Myocardial Deficiency. — Inject slowly into vein phenol,
i per cent., about 5 c.c. (=50 mg.) per kg. Stop before condition becomes
too dangerous.
Experiment n. Cardiac Dilation from Saline Infusion. — Inject normal
saline into vein. Vary speed of injection. Stop when dilation becomes too
great.
Experiment 12. Cardiac Failure from Excessive Epinephrin. — Inject
slowly into vein a i : 10,000 solution of epinephrin until heart stops. If
stoppage should not occur, follow with phenol as in Experiment 10.
QUESTIONS
(a) Describe the effects of morphin narcosis, reflexes, pain, pulse, res-
piration.
(b) Describe the phenomena of the induction of ether anesthesia.
(c) Describe the effects of curare on blood-pressure and heart.
(d) Describe the effects of excess of ether.
(e) Is ether anesthesia by insufflation a safe procedure?
(/) What are the effects and dangers of excessive intratracheal pressure?
Explain them.
(g) Describe the effects of obstruction of the air passages under ether
and under chloroform.
(h) Describe the phenomena on substituting chloroform for ether.
(i) Describe the effects of excessive chloroform on blood-pressure and
heart.
(k) Is the fall of pressure cardiac or vasomotor?
(/) What are the first danger signs?
(m) Why is chloroform insufflation more dangerous than ether?
(») Describe the effects of phenol on blood-pressure, heart, and motor
system.
(0) How does myocardial weakness modify the course of anesthesia?
(p) Describe the effects of saline infusion on the blood-pressure and
heart.
(q) Would saline infusion in collapse during anesthesia be beneficial or
harmful? Explain.
(r) Describe the effects of epinephrin on blood-pressure and heart.
(s) What is the danger of epinephrin in chloroform collapse?
(/) How would this be guarded against?
EXERCISE VII.— RESUSCITATION
If an animal is killed during anesthesia, proceed at once to resuscitation.
Experiment i. Reflex Stimulation. — Stretch the anal sphincter. If this
is not immediately effective, proceed to
tment
CHAP. XLIII VASOMOTOR DRUGS ^j,,
Experiment 2. Artificial Respiration.— If this does not succeefl proik£ityy er
perform of
Experiment 3. Cardiac Massage. — /. c.. Strong, rapid, rhythmic compres-
sion of tin- thorax (rate of at least 80 per minute). This must bp done 4e9f °
vigorously. Observe on the tracing that an artificial circulation um I
kept up in this manner. If the animal does not revive in two minutes,
continue the procedure, but at the same time proceed to
Experiment 4. Intravenous Epinephrin. — i mg. (i c.c. of i : 1000)
washed in with 50 c.c. of N. S. This aids resuscitation by stimulating the
heart and blood-vessels. If it does not succeed in two or three minutes,
proceed to
Experiment 5. Epinephrin into Carotid. — Connect cardiac end of carotid
with pressure bottle containing N.S., placed 3 or 4 feet above the table.
With a syringe inject i mg. (i c.c. of i : 1000) epinephrin into the connecting
rubber tube, while the saline is flowing in. The massage must be continued.
The epinephrin, administered in this way, reaches the coronary vessels
more directly.1
QUESTIONS
(a) Record the success or failure of these different methods of resuscita-
tion.
(b) Explain their mechanism.
EXERCISE Vin.— (OPTIONAL) MORPfflN-SCOPOLAMIN-ETHER
SYNERGISM ON MICE
W. Straub, 1912, Zs. biol. Technic., 2, 277; Fuehner, Deut. Med. Woch., 1913, No. 3.
EXERCISE IX.— (OPTIONAL) RESUSCITATION BY INTRAPERICARDIAL
INJECTION
Gunn and Martin, 1915, Jour. Pharmacol., 7, 31.
CHAPTER XLIII
VASOMOTOR DRUGS; TREATMENT OF CIRCULATORY COLLAPSE
H MEMBERS OF EACH GROUP)
Introduction (Interpretation of Blood-pressure). — The observation of
the blood-pressure is perhaps the most commonly used method for studying
changes in the circulation. However, it has certain limitations: tin- ordi-
nary method- permit only the observation of acute changes. These gener-
ally require toxic rather than therapeutic doses. Allowance must then be
i"or thi> fact.
In the second place, the changes in blood-pressure give only the sum of
the changes produced in ti it ion, hut do not usually >ho\v how and
where these effects take place. Changes in blood-pi M.I\ l>e either
lac or vascular. The mercury pr< a very imperfect
and often erroneou- imj>re»ion <>f the strength of the heart-beat. It i>
therefore necessary to distinguish between cardiac and \a-c ular c hanges by
:»eriment>. The cardiac effccti m.iv i,e registered with the myo-
> Guthrie, 1908, obtained better results by blocking the aorta.
266 A LABORATORY GUIDE IN PHARMACOLOGY
cardiograph. They may also be deduced from the vein pressure, oncometer,
or circulation time: these vary generally in the same direction as the arterial
pressure if the changes are cardiac; in the opposite direction if they are
vascular. However, the conclusions may be deceptive if the drug acts
unequally on different vascular areas. Vascular changes may also be dis-
tinguished by direct inspection.
Fairly definite conclusions may be drawn from the relation of the systolic
and diastolic pressure as recorded by a membrane manometer. The
diastolic changes are relatively greater with alterations of the vasomotor
tone or heart-rate; whereas the systolic changes are relatively greater with
alterations of the cardiac force or blood volume. Therefore, if (A) the
diastolic pressure rises relatively more than the systolic, this points to general
vasoconstriction or to quickened heart-rate. (B) The diastolic pressure
falls more than the systolic: general vasodilation or slower heart-rate.
(C) The systolic pressure falls more than the diastolic: cardiac weakening
or diminished blood volume (hemorrhage). (D) The systolic pressure
rises more than the diastolic : cardiac stimulation or increased blood volume
(transfusion). (Pilcher, 1915 Amer. Jour. Physiol., 38, 208.)
If the changes are cardiac it is necessary to distinguish between actions
on the cardiac muscle and on the nervous mechanisms, central and per-
ipheral.
Vascular changes may concern the arterial muscle or the vasocon-
strictor or vasodilator nervous mechanism. The vasodilator system is
only important in a few situations, which are not sufficient to affect the
general blood-pressure. It is therefore necessary to consider mainly the
vasoconstrictor nerves and the muscle.
Vasoconstriction. — The seat of the stimulation may be:
1. Central. — The drug has no effect if it is injected after destruction of the spinal cord.
The venous pressure and volume of the leg increases if the drug is injected after section of
the sciatic. (Strychnin, caffein, etc.)
The stimulation may also be reflex (counterirritants) or from convulsions or asphyxia.
These must be excluded by curare and artificial respiration.
A direct method for studying the reactions of the vasomotor center is described in
Exercise VI.
2. Peripheral. — (The drug is effective after destruction of the spinal cord.) The
stimulation may be in:
The Ganglia. — The drug does not act on excised organs. If the drug slows the stream
through excised organs, the action must be either on the endings (suprarenal) or on the
muscle-fibers (barium). The distinction between these is not easy. If the endings alone
are affected, the drug will not act on every organ, and it will fail to act after apocodein, or
after the organ has been excised for some hours. If the effect is on the muscle, it can be
obtained in all organs and for many hours after removal from the body, and after apo-
codein.
Simultaneous Action at Several Points. — The above experiments indicate only the
most peripheral structure on which the drug acts. If it affects a peripheral structure and
the center simultaneously, a positive distinction is possible only by maintaining a separate
artificial circulation through the center. By this means it has been shown that nitrites
paralyze the vasoconstrictor mechanism both centrally and peripherally. These experi-
ments, however, are so complicated that they are open to fallacies.
Vasodilation. — The paralysis may be:
1. Central. — Stimulation of the peripheral end of the splanchnic nerve raises the
blood-pressure; asphyxia, or central stimulation of the sciatic or of the cardiac depressor
does not alter the blood-pressure. The paralysis may be direct (chloral, chloroform) or
reflex (depressor stimulation, shock), or the result of extreme asphyxia or anemia. These
must be excluded.
2. Peripheral. — Stimulation of the splanchnic is ineffective. Paralysis, of the ganglia
(as by nicotin) is excluded by stimulating beyond them. If this is still effective, the action
must be on the endings, muscle, or capillaries. If it is on the endings, the effect of supra-
CHAP. XLIII
VASOMOTOR DRUGS
267
renal muscle will be abolished or diminished, but barium will still be effective. Paralysis
of the endings is produced by nitrites (probably), apocodein, large doses of ergot, etc. If
the muscle is paralyzed, even barium will fail to produce a rise.
With arsenic and some other metals there is a fall of pressure of vascular origin, but
the vasomotor mechanism responds well to direct or reflex stimulation. These drugs act
on the capillary walls. Capillary paralysis is also characterized by greater permeability
— intravenous injection of salt solution leading readily to muscular edema (Magnus, 1809).
Technical References. — Lateral pressure in different arteries, Dawson, 1006, Amer.
Jour. Physiol., 15, 244; Blood-pressure variations in normal dogs, Hoskins and Wheelan,
1014, Amer. Jour. Physiol., 34, 81; Percentilc Measurement of Vasomotor Reflexes, Porter,
ibid., 33, 373; Relation Blood-pressure and Respiration, Th. Lewis, 1908, Jour. Physiol.,
37, 213.
TECHNICAL NOTES ON VASOMOTOR NERVES
The splanchnic nerves may be stimulated by placing the electrodes,
spread fairly wide apart, about the hilus of the suprarenal gland. This
may be reached by the same incision as the kidney. To limit the stimu-
Fig. 60.— Dissection of left splanchnic nerve, dog.
In t inn ^trictly to the splanrhnirs. <>r to divide the-e nerves, practice dis-
sections are indispensable. In tin- rabbit the -planehnu- trunk- may I r
found in the thorax, about the tenth doisd vertebra, on each side of the
aorta. The left i- the more- racily found in the abdomen. It accomp.
the aorta until it terminate- in tin- l«>\\er « elia. -an.ulion. just alx)ve the left
suj)i e front of t lie aorta. The right spUnchnii i-M-|)arated
from the aorta, in tin- abdomen, by the vena cava. It terminates in the
i^lion at the level of the riiiht suprarenal irland. in front
of the vein (Figs. 60, 61; Burton-Opiz, 1908, Arch. ges. rhy>i«»l.. 123, 590).
268 A LABORATORY GUIDE IN PHARMACOLOGY
Technical References.— Tigerstedt, 2.2, 133, Jour. Physiol., 16, 163; Burton-Opitz,
Splanchnic and Renal Nerves, Arch. ges. Physiol., 123, 590; Splenic Vessels and Nerves,
ibid., 129, 190; Nerves of Portal Vein, Amer. Jour. Physiol., 36, 325; Duodenal Nerves,
ibid., 36, 203; Pulmonary Vasomotors, Cloetta and Anderes, Arch. Exp. Path. Pharm., 77,
Destruction of Nerves. — To complete the destruction of nerves, when these accom-
pany vessels, the sheath is painted with concentrated phenol (Bayliss, 1902).
Depressor Nerve. — In cats and dogs this is generally united with the
vagosympathetic. It may be stimulated by dividing both vagi and stimu-
lating the cephalic end of the mixed nerve. The results are usually satis-
factory hi cats, not in dogs.
Fig. 61. — Dissection of right splanchnic nerve, dog.
In rabbits the depressor runs separately from the vagus and sympathetic. It is the
most slender of these nerves, and lies a little to the inner side of the vagus (the largest
nerve) . It may be identified by the result of stimulation, and by its double origin from the
vagus and superior laryngeal. References: Tigerstedt, 2.4, 374.
TECHNICAL NOTES ON DESTRUCTION OF THE VASOMOTOR CENTER
The principal vasomotor center is situated in the medulla oblongata
extending from i or 2 mm. below the corpora quadrigemina downward to
within 4 or 5 mm. from the point of the calamus scriptorius, i. e., just above
the respiratory center. Subsidiary centers are also situated in the spinal
cord.
CHAP. XLIII VASOMOTOR DRUGS 269
The vasomotor center may, therefore, be practically excluded by cutting the spinal
cord between the calamus and the origin of the vasomotor nerves, which begin about the
second dorsal. If the section is made above the fifth cervical nerve, the respiration is also
arrested; if made between these regions, the chief vasomotor center is excluded, but the
respiration continues. In dogs, a knife thrust through the occipito-atlantoid membrane
will divide the medulla just about the lower limit of the vasomotor center.
It should be remembered that there are seven cervical and thirteen dorsal vertebrae,
the long spine corresponding to the first dorsal. The first pair of cervical nerves leave
through the atlas; the second pair between the atlas and the second vertebra; the third
pair between the second and third vertebrae, etc.; the eighth cervical between the last cer-
vical and the first dorsal vertebrae, etc.
Section of the Spinal Cord to Exclude the Vasomotor Center. — The
deeply anesthetized and tracheotomized dog is laid on the abdomen, with-
out tying. The neck is rendered prominent by drawing the head over a
sand-bag, block, or brick. An incision is made through the skin and muscles,
to the spine, from the occiput for a distance of 3 or 4 inches. Artificial
respiration is started. The cord is divided between the third and fourth
cervical vertebrae. This may be done without removing the vertebrae by
pushing a narrow-bladed knife between the articulations. It is more certain,
however, to expose the cord. This should be done as quickly as possible,
keeping closely to the middle line and to the bone, and the profuse hemor-
rhage controlled by packing tightly with small pledgets of cotton. The
vasomotor centers may be excluded with absolute certainty by destroying
the cord, passing a strong brass rod down the spinal canal.
The extent of the section or destruction must always be controlled by
sciatic stimulation and by subsequent autopsy.
If it is desired to paralyze the vasomotor center, the spine may be opened from the
third cervical vertebra upward, and packed with cotton saturated with 5 per cent, cocain
solution. This may again be rinsed off after a time.
Technical References for Operations on the Cord. — Tigerstedt, 2.4, 338; 3.4, 55.
Technical Notes on Study of Vascular Reactions. — Inspection of Blood-
vessels.— The vascularity of the organ (rabbit's ear, exposed intestine,
kidney, etc.) and the color of the venous blood are noted.
References. — Heinz, 2, 144; Robert, Into*., i, 232.
'.ry. — The temperature of an organ increased with the blood-flow. Refer-
ences: Tigerstedt, 2.4, 291.
Oncometry. — See page 169.
Vein Pressure. — The lateral pressure in the inferior cava is measured
by connecting tin- central end of the femoral vein with a manometer
shaped like the mercury manometer, but filled with \\ater. A little water
should IK- added from time to time to make sure that the vein is not plugged
by a clot. A t raring can be obtained by filling the mam -meter \\ith half-
•ated magnesium sulphate and connecting with a Hrodie hell
With some care a cork or hollow aluminum tloat and aluminum style can
I.e iitted directly to the manometer. A float recorder is described by
Hoskins, Gunning, and Berry, 1916, Amer. Jour. Physiol., 41, 517.
tedt, 2.4, 242, -
Peripheral Arterial Pressure (Wolf Method". Tin- femoral artery i^ linl. and a
rannul. i < <>nnei ted \\itli tin- />r rif>li< r.il end. I all in tliU \\ill itulit ate dilation «>f the
Vessels, and vi.e versa (Dossin. loii.Anh. Internal Pharmacod., 21, 447).
270 A LABORATORY GUIDE IN PHARMACOLOGY
Vein-flow. — In these methods the blood is generally defibrinated and reinjected, or
it is rendered non-coagulable by hirudin (see pages 245, 246). An outflow tube is then
introduced into the vein, terminally or by a T piece. The outflow is measured or counted;
it may also be estimated by the rate of rise of a tambour (see page 168).
Cerebral and Medullary Circulation. — This presents some special problems. Refer-
ences: Tigerstedt, 3.4, 131; isolation, Eisenbrey, 1910, Soc. Exp. Biol. Med., 7, 113; E. D.
Brown, 1915, Jour. Pharmacol., 6, 603; excised brains, see page 171; Brain volume, Tiger-
stedt, 3.4, 131.
Pulmonary Circulation. — Anderes and Cloetta, 1916, Arch. Exp. Path. Pharm., 79,
291.
Vessel Suture.— References: Abderhalden, 5, 815; Carrel, 1912, Surg. Gyn. Obst,
246; Guthrie, 1908, Jour. Amer. Med. Assoc., 51, 1658; human hair suture, ibid., 1910, 54,
349; Vessel-clamp, G. N. Stewart, 1910, ibid., 55, 647.
Transfusion. — Measurement u'ilh oiled syringe, Curtis and David, 1910, Jour. Amer.
Med. Assoc., 56, 35; Use of sodium citrate, R. Weil, 1915, ibid., 64, 425. (About i c.c. of
10 per cent, per 10 c.c. of drawn blood; its injection causes no disturbance and does not
change the coagulation time of the circulating blood.) Methods and apparatus, Jour.
Amer. Med. Assoc., 1916, 66, 1923.
Plasma pheresis (Plasma removal with return of corpuscles). — Withdrawal of blood
with re-injection of the corpuscles suspended in 0.6 per cent. NaCl. Much larger quanti-
ties can be withdrawn than in simple bleeding (Abel, Rowntree, and Turner, 1914, Jour.
Pharmacol., 5, 611).
Compression of Arteries. — Metal band, Matas and Allen, 1911, Jour. Amer. Med.
Assoc., 56, 233.
EXERCISE I.— (GROUP I) NITRITE AND EPINEPHRIN; RELATION OF
RESPONSE TO LEVEL OF BLOOD-PRESSURE
Distribution of Work. — Student B — Director and Reporter; calculates
doses; takes notes and prepares report.
Student E— Chief Operator.
Student F — Assistant Operator; weighs animal; gives injections.
Student A — Anesthetist; artificial respiration and resuscitation if neces-
sary; cleaning.
Student C — Pulse; blood-pressure tracings (pages 242-246).
Student D — Respiratory tracing (page 239).
Observations. — Heart- rate; blood-pressure tracing; respiratory tracing
(Stephen Hale experiment).
Apparatus. — Stephen Hale manometer: glass tubing 4 mm. diameter,
10 to 14 feet high, suspended vertically, with rubber connection to carotid.
The interior of the tube and connection should be well oiled, or leech extract
may be used. Mercury manometer with screw-clamp on connection for
blood-pressure tracing. Tracheal tube and tambour for respiration. Two
injection burets. Induction coil.
Animal. — Morphinized dog or cat with M. A. U. anesthetic.
Operation. — Weigh animal. Etherize and tie to board. Place cannulae
into carotid, trachea, and femoral veins with burets (one for epinephrin,
i : 1000; the other for nitroglycerin, i : 1000).
Experiment i. Epinephrin. — Open the carotid artery and let the blood
rise in the tube. When it has reached its maximum, measure the height of
the column in the systole and diastole.
Inject into vein Epinephrin, 0.05 mg. (^V c-c- of i : 1000) per kg.
Measure height of column.
Experiment 2. Amyl Nitrite. — When pressure has returned to normal,
let the animal inhale the nitrite. Measure height of column.
Operation. — Disconnect the manometer tube (and wash it before the
blood clots). Dissect the left splanchnic nerve and place on electrodes.
Connect carotid with mercury manometer and tighten screw-clamp till
CHAP. XLIII VASOMOTOR DRUGS 271
excursions are quite small. Connect trachea for respiratory tracing.
Start tracings.
Experiment 3. Splanchnic Stimulation. — While taking tracings stimu-
late splanchnic nerve with moderate shocks until the blood-pressure has
reached a maximum. Let pressure return to normal.
Experiment 4. Nitroglycerin. — Inject into vein Nitroglycerin 0.5 mg.
(0.5 c.c. of i : 1000) per kg. When the pressure has reached a minimum,
proceed to Experiment 5.
Xitroglycerin and nitrites produce considerable fall of blood-pressure
and increase of vein-pressure and oncometer (vasomotor paralysis); and
some quickening of the pulse (vagus depression). Respiration usually
increases. The effects pass off rapidly.
If the vagus was depressed before the nitrite was given — as denoted by
fast pulse — there may not be any further quickening.
Experiment 5. Splanchnic Stimulation During Nitroglycerin Fall. —
When the blood-pressure has reached its minimum under nitroglycerin
again stimulate the splanchnic until pressure ceases to rise. Let conditions
return to normal.
Experiment 6. Compression of Aorta. — Clamp aorta where it emerges
from diaphragm.
Experiment 7. Nitroglycerin During Compression of Aorta. — Leave
clamp on aorta. When pressure ceases to rise inject nitroglycerin as in
Experiment 4.
Experiment 8. Compression of Aorta During Nitroglycerin Fall. — Inject
nitioglycerin until pressure has fallen to the minimum of Experiment 4.
Then clamp aorta until pressure ceases to rise. Release aorta.
Experiment 9. Epinephrin. — When conditions have returned to normal,
inject into vein Epinephrin, 0.05 mg. (^ c.c. of i : 1000) per kg.: rise
in blood-pressure and fall in vein-pressure and oncometer (peripheral
vasoconstriction) ; slower pulse (vagus stimulation) and stronger heart
(stimulation cardiac muscle); respiration usually increased (higher blood-
pressure?). Cardiac slowing often precedes the vasoconstriction. Note
that the effects disappear rapidly.
Experiment 10. Epinephrin During Nitroglycerin. — When conditions
have returned to normal, inject nitroglycerin as in Experiment 4. When
the pressure has fallen to the minimum, inject epinephrin as in Experi-
ment 9.
Experiment u. Nitroglycerin During Epinephrin. — Inject epinephrin
very slowly, adjusting stop-cock so that a uniform, moderate rise of pres-
sure of 30 to 50 mm. is maintained. When this condition is reached, and
while the epinephrin is still running in, inject with a hypodermic syringe
into the vein rnniie< tion nitroglycerin. a> in Experiment 4. When press-
ure ha> fallen to minimum, discontinue the epinephrin and let conditions
return to normal.
Experiment 12. Nitroglycerin During Hemorrhage.- Insert cannula
into femoral artery and withdraw blood until pressure has fallen by 25 to
40 mm. Inject nit ro«jly< crin a- in l\\prriment 4.
Experiment 13. Strophanthus. ('Kan tpiiuphrin buret and through
it inject strophanthu^. i m-j < T\y c.c. of i : 100) per kg.
Experiment 14. Nitroglycerin During Strophanthus. — When pressure
Ceases to ri>e. inject nitroglycerin i -.periment 4.
272 A LABORATORY GUIDE IN PHARMACOLOGY
QUESTIONS
Describe the effects of:
(a) Epinephrin (Experiments i and 9) .
(b) Amyl nitrite and nitroglycerin (Experiments 2 and 4).
(c) Splanchnic stimulation (Experiment 3).
(d) Compression of aorta (Experiment 6) .
(e) Hemorrhage (Experiment 12).
(/) Strophanthus (Experiment 13).
How efficiently does nitroglycerin counteract rise of pressure produced
by:
(g) Compression of aorta? (Experiment 7.)
(h) Epinephrin? (Experiment n.)
(i) Strophanthus? (Experiment 12.)
(Compare these with the millimeter fall and with the level of pressure
reached in Experiment 4.)
(k) Where would the action of the nitroglycerin be located? (Epi-
nephrin acts on the myoneural junction; Strophanthus on the arterial
muscle) .
(/) Which vascular area is mainly affected by the nitroglycerin. (Com-
pare Experiments 7 and 4).
(m) How does hemorrhage affect the nitroglycerin fall? (Compare
Experiments 12 and 4.) Explain.
(n) How does nitroglycerin affect the response to splanchnic stimulation?
(Compare Experiments 3 and 5.) Explain bearing on location of nitro-
glycerin action.
(o) How efficiently does epinephrin counteract the nitroglycerin fall?
(Compare Experiments 9 and 10.) Explain bearing on location of nitro-
glycerin action.
(p) Summarize evidence as to location of nitroglycerin action.
(q) State some possible therapeutic applications of nitroglycerin.
OPTIONAL VASOMOTOR EXPERIMENTS
Experiment 15. Doses of Various Vasomotor Drugs Not Used in the Regular Experi-
ments.— Aconite: 15 mg. per kg., ineffective; 100 mg. per kg., fatal.
Berberin: i mg. per kg.
Camphor: 5 mg. per kg.
Cyanid Potassium: i mg. per kg., blood-pressure rise.
Digitalis: 50 mg. per kg., rapid therapeutic action; 100 mg. per kg., toxic.
Ergotoxin: 0.25 mg. per kg.
Ether: J to 2 c.c. of sat. sol. per kg.
Hydrastin : 5 mg. per kg.
Lactic Acid: 2 c.c. of 0.6 per cent, per kg.
Spartein: 5 mg. per kg.
Strychnin: 0.5 mg. per kg., tetanic; i mg. or over per kg., depression of vasomotor
center.
Experiment 16. Position on Blood-pressure. — Tigerstedt, 2.4, 302.
Experiment 17. Cerebral Compression. — H. Gushing, 1902, Grenzg. Med. Chir., 9,
793; Eyster, Burrows, and Essick, Jour. Exp. Med., n, 489; Tigerstedt, 2.4, 288.
Experiment 18. Depressor Stimulation with Vasomotor Drugs. — Sollmann and
Pilcher, 1912, Amer. Jour. Physiol., 30, 369.
Experiment 19. Thyroid Sensitization of Depressor and Epinephrin. — Asher and Flack,
1911, Zs. Biol., 55, 83 (one "tablet" in 10 c.c. of dilute alkali; filter; 2 c.c. of filtrate per kg.,
vein).
Experiment 20. Heating of Carotid Blood. — Stewart, 297.
Experiment 21. Clamping of Carotid Arteries; Traction on Cephalic End of Carotid. —
Sollmann and Brown, 1912, Amer. Jour. Physiol., 30, 88.
CHAP. XLIII VASOMOTOR DRUGS 273
EXERCISE II.— (GROUP II i PERIPHERAL VASOMOTOR DRUGS ON BLOOD-
PRESSURE AND KIDNEY VOLUME, WITH INSPECTION OF INTESTI-
NAL VESSELS
Distribution of Work. — Student B — Director and Reporter; calculates
doses; takes notes and prepares report.
Student E — Chief Operator.
Student F— Assistant Operator; weighs animal; gives injections.
Student A — Anesthetist; artificial respiration and resuscitation if neces-
sary; clean ing.
Student C — Pulse; blood-pressure tracings.
Student D — Kidney volume (page 169) and watch color of intestines.
Observations. — Heart-rate; blood-pressure tracing; kidney oncometer
record or tracing; color of intestines (anemia, congestion, etc.). If the on-
cometer is merely read from the manometer, the readings should be recorded
at the proper place on the tracing.
Apparatus. — Damped mercury manometer for blood-pressure tracing;
tracheal cannula; kidney oncometer with water-manometer (and recording
device, if desired). Injection buret; induction coil.
Animal. Morphinized dog, or cat with M. A. U. anesthetic.
Operation. — Weigh the animal. Etherize and tie on board. Place
cannulae into carotid, trachea, and femoral vein. Open abdomen and ar-
range kidney in oncometer. Draw out a loop of intestine for observation
(cover with warm towel). Connect carotid for tracing.
Injections. — Make all injections into vein; let conditions return as near
as possible to normal between injections.
Experiment i. Strychnin (Therapeutic Dose). — Inject Strychnin, 0.05
mg. (^V c.c. of i : 1000) per kg. Generally no noticeable result (this would
correspond to about -^ grain for a man) .
Experiment 2. Amyl Nitrite. — Administer by inhalation (see Exercise I,
Experiments 2 and 4, page 271).
Experiment 3. Epinephrin. — Inject 0.05 mg. (^ c.c. of i : 1000) per kg.
(see Exert ise I. Experiment 9, page 271).
Experiment 4. Pituitary.— Inject Pituitary Solution, o.i c.c. per kg.:
moderate but rather prolonged rise of pressure, often preceded by short
fall; intestinal vessels contract, kidney may dilate (peripheral constriction
of vessels; cardiac depression). Intestinal movements increased.
Experiment 5. Ergot.— Inject 250 mg. (i c.c. of 25 per cent.) pe
Effects variable: generally a moderate rise, which may be preceded by a
temporary fall.
During the fall the heart i> weakened and quickened, and the oncometer
:nini>he«l. The fall is therefore due to weaken ing of the heart . Ih
the rise the heart is strengthened; the oncometer may increase or remain
•nary. The onsequently due to strengthening of the heart,
with some vasoconstrict ion.
During the fall the heart is quickened and the respiration increased.
This is due to the low blood-pressure. The cardiac effect- can be repro-
duced on excised hearts and are therefore direct actions
The fall ot" pressure in not seen when the ergot is injected subcutaneously
or into the muscles.
Experiment 6. Tyramin. — Inject 2 mg. (2 c.c. of i : 1000) per kg.: ri>e
ofraood-prcssure by peripheral vasomot min, HUtamin and Cholin,
together with Ergotoxin are active constituents of Ergot).
18
274 A LABORATORY GUIDE IN PHARMACOLOGY
Experiment 7. Histamin. — Inject o.oi mg. (^ c.c. of i : 10,000) per kg.:
fall of blood-pressure.
Experiments. Cholin. — Inject 2 mg. (2 c.c. of i : 1000) per kg.: rise
of blood-pressure.
(Optional) This fall does not occur after the intravenous injection of atropin — i mg.
for cats.
Experiment 9. Cotarnin. — Inject 5 mg. (J c.c. of i : 100) per kg.: fall
of blood-pressure. (This substance has been tried as a hemostatic.)
Experiment 10. Hydrastis. — Inject 20 mg. (i c.c. of 2 per cent., filtered)
per kg. Short fall of pressure, followed by persistent rise. Both phenomena
are in part cardiac, in part vascular. The oncometer results are therefore
variable.
Experiment n. Hydrastinin.— Inject 5 mg. (i c.c. of i : 100) per kg.
Rise of pressure, mainly cardiac. (Hydrastinin is an artificial derivative
of Hydrastin, a hydrastic alkaloid; it has been suggested as a circulatory
stimulant, but has not found much application.)
Experiment 12. Nicotin. — Expose the vagus and find the smallest
stimulus which just stops the heart. Inject Nicotin, o.i mg. (yj c.c. of
i : 1000) per kg. The peristalsis is greatly increased. The respiration is
also increased and the animal may become convulsive. When the heart
has become quickened, note that stimulation of the vagus does not stop
the heart (depression of the vagus ganglion cells) . Very strong stimulation
may cause some slowing if the paralysis is incomplete.
Experiment 13. (Optional) Nicotin on Ganglia and Nerve-fibers. — Expose the superior
cervical ganglion of an anesthetized rabbit. Stimulation causes constriction of the ear
vessels and dilation of the pupil. Paint i per cent, nicotin on the nerve below the ganglion.
A stimulus applied central to this point is still effective, showing that the nerve-fibers are
not paralyzed by the poison. Paint the nicotin on the ganglion. Stimulation of the nerve
is now ineffective, showing paralysis of the ganglion.
QUESTIONS
(a) Tabulate the effects of the drugs on pulse-rate, blood-pressure,
kidney volume, and intestinal vessels.
(b) The organ volume or congestion varies in the same direction as the
blood-pressure if a change is cardiac, and inversely if it is vascular. On
this basis, state for each of these drugs whether the blood-pressure change is
cardiac or vascular.
EXERCISE m.— (GROUP III) PERIPHERAL AND CENTRAL VASOMOTOR
DRUGS ON BLOOD-PRESSURE, INTESTINAL VOLUME, AND RESPIRA-
TION: TREATMENT OF PEPTONE SHOCK
Distribution of Work. — Student B — Director and Reporter; calculates
doses; takes notes and prepares report.
Student E — Chief Operator.
Student F — Assistant Operator; weighs animal; gives injections.
Student A — Anesthetist; artificial respiration and resuscitation if neces-
sary; cleaning.
Student C — Pulse; blood-pressure tracings (pages 242-246).
Student D — Respiratory tracing (page 239) and intestinal oncometer
(page 169).
Observations. — Heart-rate; blood-pressure tracing; respiratory tracing;
intestinal oncometer, record or tracing; color of intestines (anemia, conges-
CHAP. XLIII VASOMOTOR DRUGS 275
tion, etc.). If the oncometer is merely read from the manometer, the read-
ings should be recorded at the right place on the tracing.
Apparatus. — Damped mercury manometer for blood-pressure tracing.
Tracheal cannula. Intestinal oncometer with water-manometer (and
recording device if desired). Injection buret.
Animal. — Morphinized dog, or cat with M. A. U. anesthetic.
Operation. — Weigh the animal. Etherize and tie on board. Place
cannulae into carotid, trachea, and femoral vein. Open abdomen and ar-
range intestinal loop in oncometer. Draw out a loop of intestine for ob-
servation (cover with warm towel). Connect carotid for tracing.
Injections. — Make all injections into vein; let conditions return as near
as possible to normal between injections.
Experiment i. Strychnin (Therapeutic Dose). — Inject Strychnin, 0.05
mK- (A c-c- of i : looo) per kg.: generally no noticeable result (this would
correspond to about ^ grain for a man).
Experiment 2. Sodium Nitrite.— Inject 5 mg. (^ c.c. -of 10 per cent.)
per kg. (See Exercise I, Experiment 4.)
Experiment 3. Epinephrin. — Inject 0.05 mg. (^ c.c. of i : 1000) per
kg. (See Exercise I, Experiment 9.)
Experiment 4. Alcohol. — Inject i c.c. (4 c.c. of 25 per cent.) per kg.
Experiment 5. Sodium Diethyl Barbiturate (Veronal). — Inject 0.2 gm.
(2 c.c. of 10 per cent.) per kg. (Jacobj and Roemer, 1911, Arch. exp. Path.
Pharm., 66, 241).
Experiment 6. Peptone Shock. — Inject Witte's Peptone slowly, 0.2 to
0.5 gm. (2-5 c.c. of 10 per cent.) per kg., until pressure remains below 40
mm. The oncometer also falls, but the large splanchnic veins appear con-
gested (probably loss of tone of splanchnic vessels). The condition is
probably similar to ordinary "shock."
Treatment of Shock. — Note efficiency or failure of the following pro-
cedures:
Experiment 7. Ammonia Inhalation. — Blow Ammonia vapor into nose:
little or no effect.
Experiment 8. Saline Infusion. — Inject warm Saline, 5 to 25 c.c. per kg.:
no improvement; on the contrary, increase of splanchnic congestion.
Experiment 9. Epinephrin. — Repeat Experiment 3: diminished re-
sponse.
Experiment 10. Strophanthus. — Inject i mg. (o.i c.c. of i : 100) per kg.:
fair response. When pressure ceases to rise.
Experiment n. Epinephrin During Strophanthus.— Make continuous
injection of Epinephrin: the blood-pressure can be maintained at an
effective level (Pearce and Eisenbrey, 1910, Arch. Int. Med., 6, 218).
QUESTIONS
(a) Tabulate the effects of the drugs of Experiment i to o <>n respira-
tion, pulse-rate, blood-pressure, intestinal volume, and intestinal vessels.
(b) The organ volume or congestion varies in the same dirt i turn a> the
blood-pressure if a change is cardiac, and inversely if it is vascular. On
thi> IMMS, state for each «.f these drugs whether the blood-pressure change
is cardiac <>r \a-dilar.
(c) What relations have the respiratory changes to the blood-pressure?
(d) Describe the results of the treatment of "shock" and discuss the
efficiency of the measures.
276 A LABORATORY GUIDE IN PHARMACOLOGY
TECHNICAL NOTES
Traumatic (Surgical) Shock. — The intestines of the anesthetized animal are exposed
and severely manipulated.
Toxic Shock (Diphtheria Toxin).— Dosage, etc., H. Meyer, Arch. Exp. Path., 60.
EXERCISE IV.— (GROUP IV) PERIPHERAL AND CENTRAL VASOMOTOR
DRUGS ON BLOOD-PRESSURE AND HEART (CARDIOPLETHYSMO-
GRAPH)
Distribution of Work. — Student E — Director and Chief Operator.
Student F — Assistant Operator; weighs animal; gives injections.
Student A — Anesthetist; artificial respiration and resuscitation if neces-
sary; cleaning.
Student B — Reporter; calculates doses; takes notes and prepares report.
Student C — Pulse; blood-pressure tracings.
Student D — Cardiac tracing.
Observations. — Heart-rate; blood-pressure tracing; cardioplethysmo-
gram.
Apparatus. — Damped mercury manometer for blood-pressure tracing.
Tracheal cannula. Cardioplethysmograph and insufflation (pages 259,
260). Injection buret. Induction coil.
Animal. — Morphinized dog.
Operation. — As in Chapter XLII, page 258. Also place cannula into
femoral artery.
Injections. — All intravenous. Let conditions return as near as possible
to normal between injections.
Experiment i. Strychnin (Therapeutic Dose). — Inject Strychnin, 0.05
mg. (fa c.c. of i : 1000) per kg.: generally no noticeable result (this would
correspond to about -fa grain for a man) .
Experiment 2. Nitroglycerin. — Inject 0.5 mg. (-fa c.c. of i : 100) per kg.
(See Exercise I, Experiment 4, page 271.)
Experiment 3. Hemorrhage. — Draw blood from femoral artery so as to
produce the same blood-pressure changes as with the nitroglycerin. Com-
pare results.
Experiment 4. Epinephrin. — Inject 0.05 mg. (fa c.c. of i : 1000) per kg.
(See Exercise I, Experiment 9, page 271.)
Experiment 5. Aortic Compression. — Compress aorta at diaphragm, so
as to produce the same blood-pressure changes as with the epinephrin.
Compare the results.
Experiment 6. Pituitary. — Inject Pituitary Solution, o.i c.c. per kg.
(See Exercise II, Experiment 4, page 273.)
Experiment 7. Phenol (Toxic Dose). — Inject 30 mg. (3 c.c. of i percent.)
per kg.: collapse. Pressure falls (vasomotor and cardiac paralysis, beats
fast and small (cardiac depression); respiration lessened (depression of
center) ; convulsive (stimulation spinal cord) .
Experiment 8. Chloral. — Inject 0.5 gm. (5 c.c. of 10 per cent.) per kg.:
fall of pressure, vasomotor and cardiac.
Experiment 9. Strophanthus. — Inject i mg. (o.i c.c. of i : 100) per kg.:
rise of pressure and cardiac changes similar to digitalis, but more prompt.
(See Chapter XLV, Exercise III, Experiment 4, page 286.)
Experiment 10. Arsenic. — Dissect left splanchnic and note effect of its
stimulation on blood-pressure. Also note appearance of intestine. Inject
Sodium Arsenate, 50 mg. (i c.c. of 5 per cent.) per kg.: the blood-pressure
CHAP. XLIII VASOMOTOR DRUGS 277
falls and the intestines appear congested. Stimulate the splanchnic; if
the action has not gone too far, there is a good response, showing that the
arterioles and their innervation are still effective. Compress the aorta:
the pressure rises, showing that the efficiency of the heart is maintained.
The paralysis is in the capillaries. Continue the observation of the animal,
and when the pressure falls further repeat the splanchnic and aortic tests.
The response will decrease, as in all conditions of low blood-pressure.
QUESTIONS
(a) Tabulate the effects on the blood-pressure and heart-rate, excursions,
ic and diastolic tone.
(b) In how far may the cardiac effects of each drug be the indirect
result of the blood-pressure changes? (Compare with Experiments 3
and 5.)
(c) Discuss the evidence for the location of the arsenic fall.
EXERCISE V.— GROUP V) REACTIONS OF THE VASOMOTOR CENTER
(PERFUSION METHOD)
Outline of Method. — (Sollman and Pilcher, 1910, Amer. Jour. Physiol.,
26, 233.)
The vessels of the spleen, kidney, etc., are ligated and perfused, leaving
the nerves uninjured. The circulation of the organ is thus completely
severed from that of the animal, and can only be influenced through the
vasomotor center. The vein-flow from the organ being recorded, a slowing
must correspond to central vasoconstriction, quickening of the flow to
central vasodilation.
Distribution of Work. — Student B — Director and Reporter; calculates
doses; takes notes and prepares report.
Student E— Chief Operator.
Student F — Assistant Operator; weighs animal: gives injections.
Student A— Anesthetist; artificial respiration and resuscitation if neces-
sary; cleaning.
Student C — Pulse; blood-pressure tracings.
Student D — Vein outflow.
Observations. — Heart-rate; blood-pressure tracing; outflow 1
Apparatus. — Bellows for artificial respiration. Damped mercury man-
ometer. Tra< heal rannula. Two-liter Mariotte liottle. suspended 4 feet
above animal, connected with Wolff bottle, resting in water-hath at 40° C.
Thi> in turn i- to be com ith the artery, tin- whole arrangement
being filled with Locke's fluid. Outflow tube, eonneeted with the vein,
and delivering into a "dipping bucket," connected with elect rir time-marker
on drum. Curare, j per cent.
Animal. — Morphini/.ed d<
Operation. — Weigh animal. Etherize. Tie on board. Place cannulae
in carotid, trachea, and femoral vein. Expose spleen ior kidr.ey) through
small incision. The largest artery and it- at eompanyinv: vein are n
All the remaining vessels and tissues are tied off in two masses \)\ ftl
ligatures. The reserved artery and vein are then cleaned with blunt
lie, avoiding injury to the ner\ .
The i i cannula i- next tied into the artery (again avoiding the
nerve-) and the per!iM«>n i- Parted. When the -pleen h. 'hat
flushed, the outflow rannula i> plated in the vein and Co: v ;ih the
278 A LABORATORY GUIDE IN PHARMACOLOGY
dipping bucket. Both cannulae point toward the spleen. They are fixed
in position. The tracings are now started. Should the animal struggle,
artificial respiration is started and curare injected (§ c.c. of J per cent, per
kg.)-
Injections. — These are all made into the femoral vein. Let conditions
return to normal between the experiments.
Experiments. — Determine the effects of the following procedures:
1. Asphyxia.
2. Hemorrhage, slight and severe (defibrinate the blood).
3. Reinjection of defibrinated blood.
4. Asphyxia.
5. Nitroglycerin, 0.5 mg. (fa c.c. of i : 100) per kg.
6. Epinephrm, 0.05 mg. (fa c.c. of i : 1000) per kg.
7. Strychnin (therapeutic dose), 0.05 mg. (fa c.c. of i : 1000) per kg.
8. Chloroform inhalation.
9. Caffein, 10 mg. (i c.c. of i per cent.) per kg.
10. Cevadin, 0.05 mg. (fa c.c. of i : 1000) per kg.
11. Atropin, 0.05 mg. (fa c.c. of i : 1000) per kg.
,12. Cevadin, as in 10.
13. Strophanthus, i mg. (fa c.c. of i per cent.) per kg.
14. Strophanthus, 5 mg. (0.5 c.c. of i per cent.) per kg.
QUESTIONS
(a) Tabulate the effects on heart-rate and vasomotor center.
(b) State which of the drugs owe their circulatory effects mainly to the
vasomotor center and which do not.
(c) Which drugs owe their action on the vasomotor center mainly to
asphyxia?
(d) What are the effects of low blood-pressure? How are they explained?
TECHNICAL REFERENCE
Outflow Recorder. — A simple syphon recorder is described by Gunn, 1913, Proc.
Physiol., Soc., Oct.
EXERCISE VI.— (OPTIONAL) CIRCULATION TIME
The efficiency of the circulation depends mainly on the velocity of the blood-stream,
the "mass-movement" of the blood. This may be measured in several ways, the methyl-
ene-blue method of G. N. Stewart being the simplest. A 2 per cent, solution, about j
c.c. per kg., is injected into the jugular vein. A stop-watch is used to time the interval
between its arrival at a given artery (for instance, in a loop of intestine) and its passage
from here into the corresponding vein. The recognition of the color change is greatly
facilitated by the use of transmitted light. Observations are made before and during the
actions of the drugs.
The following may be tried (intravenous injections) :
(1) Vagus stimulation.
(2) Epinephrin, 0.05 mg. per kg.
(3) Nitroglycerin, 0.5 mg. per kg.
(4) Alcohol, i c.c. per kg.
(5) Caffein, 10 and 50 mg. per kg.
(6) Strophanthus, i and 3 mg. per kg.
TECHNICAL REFERENCE
Tigerstedt, 2.4, 304.
CHAP. XLIV
CHANGES IN HEART-RATE, ETC.
279
EXERCISE VU.— (OPTIONAL) BLOOD-PRESSURE ASSAY OF SUPRARENAL
PREPARATIONS
One of the most reliable methods for estimating the strength of a suprarenal prepara-
tion consists in determining the dose required to produce a definite, moderate (30 to 60
mm.) rise of blood-pressure, and comparing this with a known preparation (about i c.c. of
i : 100,000 epinephrin per dog). Details are given in the U. S. P. Dried suprarenal gland
should contain at least i per cent, of epinephrin.
The most uniform results are obtained by pithing the brain and spinal cord (through
the orbit), dividing both vagi and sympathetics, and giving artificial respiration (Elliott,
1914, Jour. Physiol., 44, 374; R. L. Levy, 1916, Amer. Jour. Physiol., 41, 495.
TECHNICAL REFERENCES
U. S. P. IX; Jour. Amer. Med. Assoc., 57, 1149, 191 z; Jour. Amer. Phann. Assoc., i,
1305, ioi -; Pittenger, 52.
Other Methods.— See page 167.
CHAPTER XLIV
CHANGES IN HEART-RATE, ETC.
(REPORTERS: A MEMBERS or EACH GROUP)
Introduction. — The influence of the heart-rate on the filling and output
of the cardiac chambers is of great therapeutic importance.
Influence of Heart-rate on Output and Blood-pressure. — The minute-
output of the heart, and with it the blood-pressure, increases with the rate:
rapidly up to about 120 beats per minute; relatively less between 120 and
210 beats; and declines above 210 beats.
Y. Henderson, 1009 (Amer. Jour. Physiol., 23, 345), finds that the output of blood with
each heart-beat under normal conditions of the circulation depends mainly on the diastolic
filling, and that this varies inversely to the heart-rate. The minute-volume varies with
the heart-rate. The output per beat (the difference between diastolic and systolic volume)
V/ol
-30
A *°
ic
prr ^li/tM^f
/yv HO X74 ,M
HO *VO *«0 ' J™
Fl*. 62.— Relation of heart-rate to amplitude (output per beat) and minute-volume under
normal circulatory conditions (after Henderson). Tin- amplitude (shaded) corresponds to tl
ference between the diastolic and systolic volumes.
does ! ix h with pulse-rates to about 80 per minute— t<> thi- p«>int tin- tin
volnr rcascs with the rate. Above 80 the amplitude <lc. n MM •- pr
until, above .^40 beats per minute, it has fallen so much that the minut.
also decreased (Fig. 62).
280 A LABORATORY GUIDE IN PHARMACOLOGY
Control of the Heart-rate.— The rate of the heart is controlled by
inhibitory impulses of the vagus, by the augmentory impulses of the
accelerators, and by the state of the cardiac muscle. Any of these may
be affected by drugs, the nervous structures both peripherally and cen-
trally, directly and reflexly. The methods of analysis were discussed in
Chapter XXXVI, page 185.
It may be recalled that slowing by central or reflex stimulation of the
vagus will not occur if the vagi have been previously divided. Peripheral
vagus stimulation would occur after vagotomy, but would be abolished by
atropin.
(Juickcnhig by depression or inhibition of the vagus center will be re-
moved by stimulation of the vagus nerves. If the vagus endings have been
paralyzed, stimulation of the vagus trunk will be ineffective. -Stimulation
of the accelerator center may be excluded by section of these nerves.
Changes of blood-pressure influence the heart-rate mainly through the
vagus center, so that rise of pressure generally slows, and fall of pressure
quickens, the rate. The blood-pressure factor may be excluded, either by
comparing the effect with equal changes of pressure produced by hemorrhage
or aortic compression (Exercise V) , or by keeping the blood-pressure constant
by a compensating device (Bayliss, 1908, Jour. Physiol., 37, 272; Jackson,
1913, Jour. Pharmacol., 4, 291).
Dissection of Vagus Nerves. — This was described in Chapter XLL
Dissection of Accelerator Nerves and Stellate Ganglia. — (The dissection should be
practised in advance.) The dog is tied on its back, front legs drawn down. The operator
stands at the head. Median incision in neck extending an inch over the manubrium.
Cross incision on either side between first arid second rib. Isolate sternonrastoid inser-
tions and divide. Isolate pectoral insertions and divide. Clean median vein and divide
between double ligature. Pull external jugular outward. Divide internal jugular be-
tween double ligature. Follow carotids (pull in) and vagi to subclavian. Clean subcla-
vian and first two branches (vertebral and costocervical) from fat. Start oxygen and
curare and resect one or two ribs. Now isolate the nerves, beginning at left side. The
branches of the inferior cervical ganglion run, from out to in:
Small fibers.
Annulus behind.
Annulus before.
Vagus stem.
Accelerator.
Anastomosis with vagus.
Anastomosis with inferior laryngeal.
Follow the annulus to the stellate ganglion, pulling the subclavian up and aboral.
The ganglion is rather outward (inward and below from origin of vertebral artery).
Technical Reference. — Tigerstedt, 2.4, 353; Anderson, 1904, Jour. Physiol., 31, 2.
EXERCISE I.— (GROUP I) HEART-RATE ON BLOOD-PRESSURE AND
CARDIAC EXCURSIONS (CARDIOMYO GRAPH)
Distribution of Work. — Student A — Director and Reporter; calculates
doses; takes notes and prepares report.
Student D— Chief Operator.
Student E — Assistant Operator; weighs animal; gives injections.
Student F— Anesthetist ; artificial respiration; cleaning.
Student B — Pulse; blood-pressure tracings.
Student C — Cardiograph tracings.
Observations. — Heart-rate; blood-pressure tracing; myocardiograph
tracing from ventricle and, if possible, from auricle; inspection of heart.
Fast tracings should be taken at the critical points.
CHAP. XLIV CHANGKS IN HEART-RATE. ETC. 28l
Apparatus. — Damped mercury manometer for blood-pressure tracing.
Insufflation anesthesia (Chapter XL11. page 258). Myocardiograph
(Cushny, 1910, Heart. 2. i). Induction coil.
Animal.— Morphinized dog or M. A. U.1 cat.
Operation. — As in Chapter XLII, Exercise VI, page 263, but adjusting
the cardiograph instead of plethysmograph.
Injections. — All into femoral vein. Let conditions return to normal
between the experiments.
Experiment i. Weak Vagus Stimulation.
Experiment 2. Maximal Vagus Stimulation.
Experiment 3. Cevadin. — Inject 0.05 mg. (^ c.c. of i : 1000) per kg.:
marked slowing, or cardiac arrest, with prompt escape (veratrum viride acts
similarly).
Experiment 4. .Section of Vagi. — Divide both va.ni
Experiment 5. Cevadin After Division of the Vagi. — Repeat Experi-
ment 3 : no effect on heart-rate.
Experiment 6. Strophanthus. — Inject i mg. (T^ c.c. of i per cent.) per
k'j. Repeat even- ten minutes till death. (See Chapter XLV, Exercise II,
criment 4.)
Questions. — (a) Describe the effects of slowing the heart on the blood-
pressure and excursions, diastolic volume, and systolic volume of heart
(Kxperiments i, 2, and
(b) Describe the effects of tachycardia, ditto (Experiment 4).
(c) Describe the effects of cevadin (Experiment 3).
(d) On what structures does it act? (Compare Experiments 3 and 4.)
(e) Describe the effects of Strophanthus.
(/) How does the heart behave differently under Strophanthus and under
vagus stimulation?
Optional Strophanthus on Febrile Heart. — Induce fever (page 2:4) or heat the
carotid blood (page 272), and try the effect of Strophanthus.
EXERCISE II.— (GROUP II) HEART-RATE ON BLOOD-PRESSURE AND
CARDIAC EXCURSIONS (CARDIOPLETHYSMOGRAPH)
Read remarks under Introduction, page 279.
Distribution of Work— As in Kxcrci-r 1. page 280.
Observations.— Heart-rate; blood-pressure tracings; cardioplethysmo-
gram; inspection of heart.
Apparatus and Operations.— As in Chapter XLII. 1 \i-rcise VI, page
263. Induction coil.
Animal. Morphini/.ed <1<>L: or M. A. U. cat.
Injections. - All into femoral vein. Let conditions return to normal
between the exprrimc-
Experiment i. Weak Vagus Stimulation.
Experiment 2. Maximal Vagus Stimulation.
Experiment 3. Cevadin.— Inject 0.05 m i : 1000) pi
marked slowing or cardiac ih prompt escape (veratrum virid<
similarly).
Experiment 4. Spartein. mR. (} c.c «.f i per cent.) per kg.:
brief rise mre; niotc la-tin^ >!o\sin^ of rate; \\cakcnini; « ;
contractions.
> M. A. U. stands for morphin-atropin-urcthanc page 248.
282 A LABORATORY GUIDE IN PHARMACOLOGY
Experiment 5. Pilocarpin. — Inject i mg. (^ c.c. of i : 100) per kg.:
heart first slowed; later it may be quickened (peripheral vagus stimulation
and depression). Cats may show pulmonary edema.
Experiment 6. Digitalis.— Inject 50 mg. (i c.c. of 5 per cent.) per kg.,
as in Chapter XLV, Exercise III, Experiment 4.
QUESTIONS
(a) Describe the effects of slowing the heart on the blood-pressure and
excursions, diastolic volume, and systolic volume of heart (Experiments
i, 2, and 3).
(b) Describe the effects of cevadin, spartein, pilocarpin, and digitalis.
(c) Discuss whether their cardiac effects are referable simply to the
change of heart-rate.
EXERCISE in.— (GROUP III) HEART-RATE, ETC., ON BLOOD-PRESSURE
AND ORGAN VOLUME
Fast tracings may be taken at the critical points.
Distribution of the Work. — Student A — Director and Reporter; calcu-
lates doses; takes notes and prepares report.
Student D— Chief Operator.
Student E — Assistant Operator; weighs animal; gives injections.
Student F— Anesthetist; artificial respiration; cleaning.
Student B — Pulse; blood-pressure tracings.
Student C — Kidney oncometer; inspection of intestinal vessels.
Observations. — Heart-rate; blood-pressure tracing; kidney oncometer;
intestinal vessels.
Apparatus. — Damped mercury manometer for blood-pressure tracing.
Tracheal cannula. Oncometer. Induction coil.
Animal. — Morphinized dog or M. A. U. cat.
Operation. — Weigh animal. Etherize. Tie on board. Place cannula
in carotid, trachea, and femoral vein. Expose kidney and arrange in on-
cometer. Draw out loop of intestine for inspection. Start tracings.
Injections. — All into vein.
Experiment i. Weak Vagus Stimulation.
Experiment 2. Maximal Vagus Stimulation.
Experiment 3. Cevadin. — Inject 0.05 mg. (f0 c.c. of i : 1000) per kg.:
marked slowing or cardiac arrest, with prompt escape (veratrum viride acts
similarly).
Experiment 4. Dog* s Urine.1 — Inject about 3 c.c.: large fall of blood-
pressure.
Experiment 5. Ouabain (Crystallized Strophanthus) . — Inject 0.05 mg.
(^jV c.c. of i : 1000) per kg. Repeat every ten minutes till death. (See
Chapter XLV, Exercise III, Experiment 4.)
QUESTIONS
(a) Describe the effects of cardiac slowing on the blood-pressure, kidney
volume, and large intestinal veins (Experiments i and 2). Explain.
(b) Describe the effects of cevadin, urine, and ouabain on these functions
and on the heart-rate.
(c) Explain their actions.
1 Urine Depressor Substances. — Pearce and Eisenberg, 1910, Amer. Jour. Physiol., 26, 26.
Fecal Depressor Substances —Wallace and Sturtevant, 1914, Soc. Exp. Biol. Med., n, 114.
CHAP. XLIV CHANGES IN HEART-RATE, ETC. 283
EXERCISE IV.— (GROUP IV) HEART-RATE ON BLOOD-PRESSURE AND
URINE FLOW
The flow of urine depends largely on the flow of blood through the kidneys
and may, therefore, be influenced by the circulation.
Distribution of Work. — Student A — Director and Reporter; calculates
doses; takes notes and prepares report.
Student D— Chief Operator.
Student E — Assistant Operator; weighs animal; gives injections.
Student F — Anesthetist; artificial respiration; cleaning.
Student B — Pulse; blood-pressure tracings.
Student C — Urine flow; inspection of kidney.
Observations. — Heart-rate; blood-pressure tracing; urine flow (this may
be counted or registered with an automatic drop recorder); inspection of
kidney; color of kidney substance and of renal vein.
Apparatus. — Drum; manometer; ureter cannula; induction coil; injection
buret.
Animal. — Morphinized dog or M. A. U. cat.
Operation. — Weigh, etherize; cannulae in carotid, tracheal and femoral
vein. Expose kidney for observations. Insert ureter cannula.
Injections. — All into vein.
Experiment i. Weak Stimulation of Vagus.
Experiment 2. Maximal Stimulation of Vagus.
Experiment 3. Cevadin. — Inject 0.05 mg. (-fa c.c. of i : 1000) per kg.:
marked slowing or cardiac arrest, with prompt escape (veratrum viride acts
similarly).
Experiment 4. Atropin. — Inject 0.05 mg. (-fa c.c. of i : 1000) per kg.:
heart quickens; moderate rise of pressure. Try efficiency of vagus stimula-
tion negative.
Experiment 5. Barium Chlorid. — Inject 20 mg. (2 c.c. of i per cent.) per
k.n.. and repeat every ten minutes until death: the effects on the heart re-
semble those of digitalis, but the vasoconstriction is much more prominent
and the pressure rises very high. The urine, however, is decreased, the
renal arteries being also constricted. The intestines show violent peristalsis.
This and the vasoconstriction are due to direct stimulation of the unstriped
muscle.
QUESTIONS
(a) Describe the effects of cardiac slowing on the blood-pressure and
kidney (Experiments i and 2).
(b) Ditto for cardiac quickening (Experiment 4).
(c) Describe the effects of cevadin, atropin, and barium.
(d) In how far are their effects explained by changes of heart-rate?
EXERCISE V.— (GROUP V) HEART-RATE AND RESPIRATION AS INFLU-
ENCED BY BLOOD-PRESSURE
Changes in heart-rate are sorm-tinu^ merely indirect results of ch.
of blood-pressure. Fast tracings should be taken at the critical point-
Distribution of Work. -Student A— Director and Report IT; tabulates
doses; takes notes and prepares report.
Student D— Chief Operator.
Student E — Assistant Operator : \v« -vj]\< animal; uivt -s injections.
Student F— Anesthetist ; artiiuial ropitation; cleaning.
Student B — Pulse; blood-pressure tracings.
Student C — Respiratory tracings.
284 A LABORATORY GUIDE IN PHARMACOLOGY
Observations. — Heart-rate; blood-pressure tracing; respiratory trarir j.
Apparatus. — Manometer; respiratory tambour; injection buret; drum.
Animal. — Morphinized dog or M. A. U. cat.
Operation. — Weigh. Etherize. Cannulae in carotid, trachea, and
femoral artery and vein. Small incision in abdomen to permit insertion of
finger to compress aorta near diaphragm.
Injections. — All into vein.
Experiment i. Nitroglycerin. — Inject 0.5 mg. (7V c.c. of i per cent.) per
kg. Pay particular attention to heart-rate. (See Chapter XLIII, Exercise
I, Experiment 4.)
Experiment 2. Hemorrhage. — Bleed animal so as to imitate the nitrite
fall of pressure.
Experiment 3. Nitroglycerin and Compression of Aorta. — Inject as in
Experiment i, but keep blood-pressure level by appropriate compression
of the aorta.
Experiment 4. Epinephrin. — Inject 0.05 mg. (?\ c.c. of i : 1000) per
kg. Pay particular attention to the heart-rate. (See Chapter XLIII,
Exercise I, Experiment 9.)
Experiment 5. Compression of Aorta. — Compress aorta so as to imitate
the epinephrin rise.
Experiment 6. Strophanthus and Hemorrhage. — Inject i mg. (TV c.c.
of i per cent.) per kg. Bleed when necessary to keep blood-pressure level
(compare with Exercise III, Experiment 5). Repeat every ten minutes till
death.
QUESTIONS
(a) In how far -may the nitrite tachycardia be explained by fall of blood-
pressure. (Compare Experiments i, 2, and 3.)
(b) Can the epinephrin slowing be explained by rise of blood-pressure?
^(Compare Experiments 4 and 5.)
(c) Can the strophanthus slowing be explained in this way? (Experi-
ment 6.)
CHAPTER XLV
MYOCARDIAL DEPRESSANTS AND TONICS
EXERCISE I.— (GROUP I) CARDIAC DEPRESSANTS ON BLOOD-PRESSURE
AND ORGAN VOLUME
(REPORTERS: F MEMBERS OF EACH GROUP)
Distribution of Work.— Student C— Chief Operator.
Student D — Assistant Operator; weighs animal; gives injections.
Student E — Anesthetist; artificial respiration; cleaning.
Student F — Director and Reporter; calculates doses; takes notes and
prepares report.
Student A — Pulse; blood-pressure tracing.
Student B — Oncometer; inspection of intestinal vessels.
Observations. — Heart-rate; blood-pressure tracing; kidney or spleen
oncometer; intestinal vessels. Draw out loop of intestine for inspection.
Start tracing.
Animal. — Morphinized dog or M. A. U. cat.
CHAP. XLV MYOCARDIAL DEPRESSANTS AND TONICS 285
Apparatus. — Damped mercury manometer; oncometer; injection buret.
Operation. — Weigh ; etherize ; tie on board. Cannulae in carotid, trachea,
and femoral vein. Expose kidney or spleen and place in oncometer.
Injections. — All into vein.
Experiment i. Aconite (Therapeutic Dose). — Inject 5 mg. (^ c.c. of
10 per cent.) per kg.: slight slowing of the heart (stimulation of vagus
centers) or no effect. Respiration increased (stimulation of center).
Experiment 2. Antipyrin. — Inject 100 mg. (i c.c. of 10 per cent.) per kg.
This illustrates the direct collapse action of "coal-tar" antipyretics.
Experiment 3. Phenol. — Inject 50 mg. (5 c.c. of i per cent.) per kg.
(See Chapter XLIII, Exercise IV, Experiment 7.)
Experiment 4. Veratnim.— Inject 5 mg. (^ c.c. of 10 per cent.) per kg.
(See Chapter XLIV, Exercise I, Experiment 3.)
Experiment 5. Aconite (Toxic Dose). — Inject 100 mg. (i c.c. of 10 per
cent.) per kg. : the heart is first slowed and strengthened (stimulation of
vagus and myocardium) ; then weak and rapid (paralysis of vagus) ; then
very irregular (overstimulation of myocardium); goes into delirium cordis
and stops. The action may require half an hour.
Experiment 6. Chloroform Rigor. — Inject some chloroform into the
peripheral end of one femoral artery : this causes immediate rigor of this leg.
QUESTIONS
(a) Describe effects of the drugs on blood-pressure and organ volume,
and intestinal vessels.
(b) How far are the effects cardiac? Explain.
(c) In what pathologic conditions would cardiac depressants be useful?
(</) How could one treat cardiac collapse?
TECHNICAL REFERENCES
Heart Weight. — Joseph, 1908, Jour. Exp. Med., 10, 521; Cardiac Stimulation, Tiger-
stedt, 2.4, 335; Reflexes, ibid., 374; Sounds, ibid., 195.
EXERCISE II.— (GROUP H) CIRCULATORY DRUGS ON ARTERIAL AND
VEIN PRESSURE
The vein pressure is mainly an index of the efficiency of the circulation.
It tends to rise when the heart is depressed; it t'ends to fall when the circula-
tion is improved (Capps and Mattht
Distribution of Work. — Student F — Director and Reporter; calculates
doses; takes notes and prepares report.
Student C— Chief Operator.
Student D — Assistant Operator; weighs animal: irivcs injections.
Student E — Anesthcti.-t ; artificial respiration; cleaning.
Student A — Pulse; blood-pressure tracing.
Student B — Vein pressure-.
Observations. -II ; blood-pressure i i in pie-Mire read-
(after each experiment let some water run into tin- \ein-manometer to
tlu>h blood out of the cannula). Transfer the readings to the proper places
on thi
Apparatus. Blood pressure; water manometer for vein; induction foil;
injection l»u:
Animal. — Morphinized dog.
286 A LABORATORY GUIDE IN PHARMACOLOGY
Operation. — Weigh, etherize, tie on board. Isolate vagus and place on
thread. Cannulae in carotid, trachea, and both femoral veins (cardiac end).
Connect one vein with water-manometer.
Injections. — Intravenous.
Experiment i. Weak Vagus Stimulation. — Rise of vein pressure.
Experiment 2. Maximal Vagus Stimulation. — Rise of vein pressure.
Experiment 3. Nitroglycerin. — Inject 0.5 mg. (^V c.c. of i per cent.)
per kg.: fall of vein pressure. (See Chapter XLIII, Exercise I, Experi-
ment 4, p. 271.)
Experiment 4. Epinephrin. — Inject 0.05 mg. (-jV c.c. of i : 1000) per
kg. : vein pressure may rise.
Experiment 5. Ergot. — Inject 250 mg. (i c.c. of 25 per cent.) per kg.
(See Chapter XLIII, Exercise II, Experiment 5, p. 273.)
Experiment 6. Barium Chlorid. — Inject 20 mg. (2 c.c. of i per cent.) per
kg. (See Chapter XLIV, Exercise IV, Experiment 5, p. 283.)
QUESTIONS
(a) Describe the effects of cardiac inhibition on vein pressure (Experi-
ments i and 2).
(b) Describe the effects of the drugs on arterial and venous pressure.
(c) Which of the drugs would be useful, and which harmful, in dilation
of the right heart or in venous hemorrhages?
EXERCISE III.— (GROUP III) CARDIAC STIMULANTS AND DEPRESSANTS
ON CARDIOMYOGRAM
Distribution of Work. — Student F — Director and Reporter; calculates
doses; takes notes and prepares report.
Student C— Chief Operator.
Student D — Assistant Operator; weighs animal; gives injections.
Student E — Anesthetist; artificial respiration; cleaning.
Student A — Pulse; blood-pressure tracing.
Student B — Cardiograph tracings.
Observations. — Heart-rate; blood-pressure tracing; myocardiograph
tracing from ventricle and, if possible, from auricle; inspection of heart.
Apparatus. — Damped mercury manometer for blood-pressure tracing.
Insufflation anesthesia (pp.. 258, 259). Myocardiograph (Cushny, 1910,
Heart, 2, i). Induction coil.
Animal. — Morphinized dog or M. A. U. cat.
Operation. — As in Chapter XLII, Exercise V., adjusting the cardio-
graph instead of plethysmograph.
Injection. — All into femoral vein. Let conditions return to normal be-
tween the experiments.
Experiment i. Caffein (Therapeutic Dose). — Inject 10 mg. (i c.c. of
i per cent.) per kg: increase of rate and excursions.
Experiment 2. Chloroform. — Let animal inhale Chloroform until there
is a marked fall of blood-pressure: cardiac depression.
Experiment 3. Spartein. — Inject 5 mg. (\ c.c. of i per cent.) per kg.
(See Chapter XLIV, Exercise II, Experiment 4, p. 281.)
Experiment 4. Digitalis. — Inject 50 mg. (i c.c. of 5 per cent.) per kg.
Therapeutic stage of digitalis action: Heart slowed, beats stronger (stimula-
tion of cardiac muscle and vagus) ; blood-pressure high (cardiac effect and
vasomotor stimulation); respiration increased (stimulation of center).
CHAP. XLV MYOCARDIAL DEPRESSANTS AND TONICS 287
When this action has been observed (waiting twenty minutes if necessary),
repeat the injection every fifteen minutes until death. Toxic stage of
digitalis: the effects of toxic doses of digitalis on the circulation are
extremely irregular, and may vary from moment to moment. The rate is
generally increased, but may be slowed at times. The irregularities usually
occur in groups; these are partly due to the influence of respiration (the
reflex excitability of the vagus being heightened), partly to arhythmia of
the auricles and ventricles. The effects are based on an increased excita-
bility of the cardiac muscle with systolic tendency, and on irregular activity
of the vagus. Death occurs suddenly, sometimes by vagus stimulation,
but more commonly by delirium cordis, the result of overstimulation of the
heart. The blood-pressure may remain high until the end, or it may fall,
according to the output of the heart and the persistence of the vasocon-
striction.
Experiment 5. Caffein Rigor. — Inject 10 c.c. of i per cent. Caffein into
the peripheral end of the femoral artery. Observe that this leg goes into
rigor before the other (drug rigor) .
QUESTIONS
Describe the effects of the drugs on the heart-rate, excursions, diastolic
and systolic volume.
EXERCISE IV.— (GROUP IV) CIRCULATORY DRUGS ON PRESSURE IN
PULMONARY ARTERY
The pressure in the pulmonary artery is determined mainly by the
pressure in the right ventricle, and is, therefore, proportional to the vein
pressure. It may also be influenced by the state of the pulmonary arte-
rioles, but this is usually a minor factor.
The pressure in the pulmonary artery may therefore rise by cardiac
insufficiency, by pulmonary vasoconstriction, or by extensive dilation of
the systemic vessels. Decrease of pressure has the opposite explanations.
Distribution of Work. — Student F — Director and Reporter; calculates
doses; takes notes and prepares report.
Student C— Chief Operator.
Student D — Assistant Operator; weighs animal; gives injections.
Student E — Anesthetist; artificial respiration; cleaning.
Student A — Pulse; blood-pressure tracing.
Student B — Tracings from pulmonary artery.
Observations. -Heart-rate; tracings of pressure in carotid and pul-
monary artery.
Apparatus. Two mercury manometers, writing above each other. In-
duction coil. Injection buret.
Animal. Morphi nixed dog.
Operation. — Weigh, etheri/e. tie on board. Place vagus on thread.
Cannula- in carotid, trachea, and femoral vein. Start artificial respiration.
Open chest as described on pp. 258, 259. Place cannula into cardiac end
of pulmn: i a lobe of the lung. Connect for tracings.
Injections. Intravet
Experiment i. Weak Vagus Stimulation.
Experiment 2. Maximal Vagus Stimulation.
Experiment 3. Nitroglycerin. Inject 0.5 mg. (fa C.C. of I per cent.) per
kg. (See Chapter X I.I 1 1 . 1 \ercise I, p. 271.)
288 A LABORATORY GUIDE IN PHARMACOLOGY
Experiment 4. Epinephrin. — Inject 0.05 mg. (^ c.c. of i : 1000) per kg.
(See Chapter XLIII, Exercise I, Experiment 9, p. 271.)
Experiment 5. Ergot. — Inject 250 mg. (i c.c. of 25 per cent.) per kg.
(See Chapter XLIV, Exercise IV, Experiment 5, p. 273.)
Experiment 6. Strophanthus. — Inject i mg. (^ c.c. of i per cent.) per
kg. Repeat every ten minutes till death. (See Chapter XLV, Exercise III,
Experiment 4, p. 286.)
QUESTIONS
(a) Describe and explain the effects of cardiac slowing on the pressure
in the pulmonary artery (Experiments i and 2) .
(b) Describe and explain the effects of the drugs on the carotid and
pulmonary arterial pressures, and on the heart-rate.
(c) Which of these drugs would be useful, and which harmful, in dilation
of the right heart?
(d) Ditto as to hemorrhage from rupture of a pulmonary artery?
EXERCISE V.— (GROUP V) CARDIAC DRUGS ON CARDIOPLETHYSMOGRAM
Distribution of Work. — Student F — Director and Reporter; calculates
doses; takes notes and prepares report.
Student C— Chief Operator.
Student D — Assistant Operator; weighs animal; gives injections.
Student E — Anesthetist; artificial respiration; cleaning.
Student A — Pulse; blood-pressure tracing.
Student B — Tracing from cardioplethysmograph.
Observations. — Heart-rate; blood-pressure tracings; cardioplethysmo-
gram; inspection of heart.
Apparatus and Operations. — As in Chapter XLII, Exercise VI, p. 263.
Induction coil.
Animal. — Morphinized dog.
Injections. — Intravenous.
Experiment i. Asphyxia and Recovery. — Arrest the flow of air. When
the heart is materially weakened, restore the flow.
Experiment 2. Strychnin (Therapeutic Dose). — Inject 0.05 mg. (^ c.c.
of i : 1000) per kg. (See Chapter XLIII, Exercise II, Experiment i, p. 273.)
Experiment 3. Potassium Chlorid. — Inject 10 mg. (i c.c. of i per cent.)
per kg. Repeat every ten minutes if necessary. The heart will be somewhat
weakened, slowed, and irregular (the pressure falling) and will stop rather
suddenly (paralysis of cardiac muscle). (Magnesium produces similar
effects.)
Experiment 4. Camphor. — Inject 5 mg. (J c.c. of i per cent, in 40 per
cent, alcohol) per kg.: usually little effect.
Experiment 5. Veratrum. — Inject 5 mg. (^ c.c. of 10 per cent.) per kg.
(See Chapter XLIV, Exercise I, Experiment 3, p. 281.)
Experiment 6. Strophanthus. — Inject i mg. (rV c.c. of i per cent.) per
kg. Repeat every ten minutes till death. (See Chapter XLV, Exercise
III, Experiment 4, p.- 286.)
QUESTIONS
(a) Describe the effects of the procedures and drugs on the blood-
pressure, heart-rate, excursions, and systolic and diastolic volume.
(b) Which of the drugs might serve as cardiac stimulants, as cardiac
depressants, and which are indifferent?
CHAP. XLVI INTESTINAL OSMOSIS-DIURESIS 289
ril.U'TKk XLVI
INTESTINAL OSMOSIS-DIURESIS—TREATMENT OF ACUTE
CARDIAC LESIONS
(REPORTERS: E MEMBERS OF EACH GROUP)
Introduction (Effects of Drugs on the Kidney). — The physiology and
pharmacology of the kidneys differ conspicuously from that of the typical
glands, such as the salivary: The kidney is not markedly affected by the
usual glanduar stimulants and depressants, such as pilocarpin and atropin.
It functionates quite well when the nervous connections are divided. Its
activity is most intimately connected with the state of the circulation. The
quantity of urine is influenced mainly by the nitration pressure, i. e., the
difference between the pressure in the glomerular capillaries and in Bowman's
capsule (cf. Chapter XXXV). This is determined by the systemic circu-
lation, by the state of the vessels within the kidney, and by the viscidity
of the blood. There is evidence that the kidneys possess an active vaso-
dilator as well as a constrictor mechanism. The composition of the urine
cannot be explained by a simple nitration theory. It necessitates the ac-
ceptance of unexplained forces. The changes occur by reabsorption and
also by secretion.
The mechanism of urine secretion may be explained by several alternative theories,
none of which is positively established to the definite exclusion of the others. The fol-
lowing working theory furnishes the most simple explanation of the phenomena: A |>
ical nitration of urine occurs in the glomeruli. The filtrate probably corresponds
protein-free plasma. The quantity of the filtrate depends mainly on the filtration pressure.
During the passage of the glomerular fluid through the urinary tubules a series of
changes occur by the operation of powerful forces which cannot yet be explained on a
physical basis. These cause the reabsorption of certain constituents and the secretion of
others. The extent of these changes is indicated by the departure of the composition of
the final urine from that of the protein-free blood plasma. It varies inversely to the rate
of urine flow (a more rapid flow leaving loss time for these changes). It is also influenced
by the composition of the blood, but in a manner which is not fully understood.
The absorption involves mainly the water and chlorids; to a less extent the sulphates
and phosphates; urea being the least absorbable constituent.
The secretion bears on the uric acid, certain pigments, and probably a variable pro-
portion of the urea and of other urinary constituents.
Diuretics (drugs which increase the urine flow) may be grouped into the following
(. la-.-e-:
Digitalis. — Acts by increasing the filtration pressure, through increased output of the
heart, with stronger pulse-pressure; through lessened venous pressure; through theabsorp-
tion of effusions, producing hydremic plethora. The diuretic tendcmy may be counter-
acted by constriction of the renal arterioles. It i>. therefore, but little diuretic in health,
y so in . ardiac disease, where the condition- l»r it- favorable a. lion are pr>
Irritant Diuretics. -Volatile oils, calomel, alcohol, etc.; probably some of th<
acids, and alkalic- -mall do-.es in. reasr the va->, ularity and thereby the tilt ration pressure,
possible that they also stimulate the secreting iger doses cause stasi- and
injury t<> the < ell-, and consequently lessened output of urine, with albuminuria, casts,
and eventually anuria.
Irritant diureti< - should not be used in nephi;1
Saline diuretics, including all sub-tames \\hi.h act by salt-action (watt-
salt solution-, glucose, urea, etc.).— These produce kydrtmic />/<•//; :ilutc
the blood. This in. rea-e- the nitration pressure by i: ;he total <|uantity of fluid;
by lessening the \i-. idity and thereby red in inn fri. tion in tin and < apili
the lessened viscidity also reduces the filtration re-i-tan. e. Stronger solutions further
in. re.t-e the tiltrat i'«n pressure by osnioti. -hrinkage of the renal .ells. It is possible that
some of these substances also stimulate the se. ret ing • elU ,,r depress the reab
ulant Diuretics (Can .eophyllin Thou in). T!
re. tly on the kidne\ ,!.iti.>n of the \c--el-. probably by shrinkii
cells, and i Itration pressure; but this is JM
19
2QO A LABORATORY GUIDE IN PHARMACOLOGY
of the diuresis. This is thought by some to involve a depression of the reabsorbing func-
tion; but it is more likely that they act by stimulating the secretory cells.
Drugs which constrict the vessels (suprarenal, barium, etc.) lessen the output of
urine, the resistance in the afferent arterioles being increased more than the general blood-
pressure. The effect of vasodilators is variable, according to whether they act more power-
fully on the systemic or on the local vessels. In excised kidneys, vasoconstrictor drugs
always lessen the urine flow, while vasodilators (cyanids) increase it.
TECHNICAL NOTES
Collection of Urine. — In operated animals cannulae are tied into the
ureters (taking care that these are not kinked) ; or irr rabbits, into the bladder
(see Chapter XXXV). In survival experiments a permanent bladder
fistula may be established (Schwarz and Wiechowski, 1914, Zbl. Physiol.,
28,440).
Diuretic Factors. — In exact experiments the urine flow is referred to the weight of the
animal, v. Schroeder selects the surplus excretion per 100 gm. of animal, calculated usu-
ally for one hour. Sollmann's factor relates to the maximal rate of secretion, being the
maximum number of cubic centimeters of urine secreted in forty consecutive minutes
per kilo of animal (Amer. Jour. Physiol., 1903, 9, 454).
TREATMENT OF CARDIAC LESIONS
The acute lesions produced experimentally are not strictly analogous in
their effects to the usual chronic clinical lesions. However, the principles
illustrated in these experiments are fairly applicable to both.
TECHNICAL REFERENCES TO CARDIAC LESIONS
General Technic. — Rosenbach, Arch. exp. Path., 9, i; Emerson, 1907,
Experimental Pathologic Lesions, N. Y. Med. Jour., April 20.
Temporary Valvular Lesions. — Wiggers and Du Bois, 1913, Soc. Exp.
Biol. Med., 10, 87.
Aortic Insufficiency. — Acute, Zollinger, 1909, Arch. exp. Path. Pharm.,
61, 193; Hasenfeld and Romberg, 1897, Arch. exp. Path., 37, 333.
Mitral Stenosis. — Hirschfelder, 1908, John Hopkins, Hosp. Bui., 19, 319.
Myocarditis. — Fleisher and Loeb, Arch. Int. Med., Feb., 1909; ibid.,
1910, 6, 427 (Epinephrin with Spartein or Caffein).
Experimental Surgery of Heart. — Werelius, 1914, Jour. Amer. Med.
Assoc., 63, 1338.
Electrocardiograms. — Tigerstedt, 2.4, 203; Interpretation, Eyster and
Meek, 1913, Arch. Int. Med., u, 204; Pardee, 1914, Jour. Amer. Med.
Assoc., 62, 1311; Waller, 1914, Harvey Lectures, p. 17. Protection of
string galvanometer against external electric disturbances, H. B. Williams,
1916, Amer. Jour. Physiol., 40, 230.
Pressure in Cardiac Cavities. — Tigerstedt, 2.4, 205; Heinz, i, 850.
Movements of Cardiac Valves. — Dean, 1915, Soc. Exp. Biol. Med., 13, 5.
DISTRIBUTION OF WORK
Student E — Director and Reporter; calculates doses; takes notes and
prepares report.
Student B— Chief Operator.
Student C — Assistant Operator; weighs animal; gives injections.
Student D — Anesthetist; artificial respiration; cleaning.
Student F — Pulse; blood-pressure tracing.
Student A — All other observations.
Animals. — Morphinized dogs.
Injections. — All intravenous.
nc STENOSIS
T °f
fliw (thi °r01
uepartmenl
CHAP. XLVI INTESTINAL OSMOSIS-DIURESIS 29»f
EXERCISE I.— (GROUP I) FIRST PART: DIURETICS; URINE FLOW, BLOOD-
PRESSURE, AND RESPIRATION; SECOND PART: AORTIC STENOSIS
WITH CARDIOPLETHYSMOGRAM
FIRST PART: DIURESIS
Observations. — Heart-rate; blood-pressure tracing; urine
be counted or registered with an automatic drop recorder);
kidney; color of kidney substance and of renal vein. Respiratory tracing.
Apparatus. — Manometer; ureter cannula; injection buret; respiratory
tambour; drum induction coil.
Operations. — Weigh; etherize; cannulae in carotid, trachea, and femoral
vein. Expose kidney for observations. Insert ureter cannula. Connect
tracheal tambour for respiratory tracing.
Experiment i. Sulphate Diuresis. — Inject 25 c.c. per kg. of warm sodium
sulphate (2.5 per cent, of dried or 5 per cent, of crystals). Collect the urine
after a few minutes. Rise of blood-pressure, stronger and usually slower
heart, increased oncometer and respiration. The effect is usually short,
and may be small, especially if the animal is in good condition. (Stimu-
lation of the medullary centers and cardiac muscle by the increased quantity
of blood, and by salt action.) The urine flow is promptly increased, and
remains high for a considerable time (dilution of blood, lessened viscidity,
increased quantity of blood in vessels, "hydremic plethora")- Note that
the carotid pressure is not increased sufficiently to account for the diuresis.
The volume of the kidney increases.
Some animals do not show any diuresis, especially if the kidneys have
been injured. Should this be the case, the ureter observations may be
abandoned, and replaced by myocardiogram, oncometer, or respiratory
tracings.
Test the urine for chlorids (HNOa + AgNO3), comparing it with the
original bladder-urine. The chlorid has almost disappeared (due to dilu-
tion of the plasma; the chlorid could be made to reappear by the injection
of sodium nitrate, iodid, bromid, or sulfocyanid. These act probably by
liberating the "combined" chlorid of the plasma).
The hypodermic or intravenous injection of normal saline solution or
the drinking of water increase the diuresis in the same manner as the sul-
phate solution. The latter would not be diuretic by mouth, as it is but
imperfectly absorbed.
Experiment 2. Epinephrin. — Inject 0.05 mg. (•£$ c.c. of i : 1000) per kg.
(See Chapter XLIII, Kxerdse I, Kxperiment 9, p. 271.)
Experiment 3. Spartein. — Inject 5 mg. (J c.c. of i per cent.) per kg.
(See Chapter XLIV, Exercise II, Experiment 4, p. 281.)
SECOND PART: AORTIC STENOSIS
Observations, Apparatus, and Operation. — (See Chapter XLII, Exercise
VI. p. *
Experiment 4. Aortic Stenosis.— Place screw-damp on aorta, as near
as possible to it- (.riirin. and tighten while taking tradnu'. ><> that the pulsa-
tions of the manometer are materially rediu ed. hut not abolished.
Experiment 5. Weak Vagus Stimulation.
Experiment 6. Strong Vagus Stimulation.
Experiment 7. Saline Infusion. — Inject slowly warm N. S., 25 c.c. per kg.
Experiment 8. Strophanthus.1 — Inject Strophanthus as in Chapter
XLIV, Exercise I, Experiment 6, p. 281.
1 Aortic Compression and Strophanthin, de Heer, 1912, Arch. get. Physiol., i .$
2Q2 A LABORATORY GUIDE IN PHARMACOLOGY
QUESTIONS
(a) Describe and explain the effects of the procedures of Experiments i
to 3 on the blood-pressure and ureter flow.
(b) Which of these would be useful in dropsy?
(c) Which in uremia?
(d) Describe and explain the effects of aortic stenosis.
(e) How is this modified by the procedures of Experiments 5 to 8?
(/) Which of these would be useful, which harmful, and which indiffer-
ent?
EXERCISE II.— (GROUP II) FIRST PART: URINE FLOW. SECOND PART:
HYDROPERICARDIUM
FIRST PART: URINE FLOW
Observations. — Heart-rate; blood-pressure tracing; urine flow (this may
be counted or registered with an automatic drop-recorder); inspection of
kidney; color of kidney substance and of renal vein.
Apparatus. — Drum; manometer; ureter cannula; drop-recorder; injection
buret; induction coil; dish, rod, funnel and strainer for defibrinated blood.
Operation. — Weigh; etherize; cannulae in carotid, trachea, and femoral
vein. Expose kidney for observation. Insert ureter cannula.
Experiment i. Absorption of Sodium Chlorid and Magnesium Sulphate.
—Make a 2-inch incision in linea alba, draw out a loop of intestine, and
ligature it in two places, about 25 cm. apart. Make an opening just inside
one of the ligatures. Strip the piece of intestine of its contents, insert the
end of a funnel into the opening, and allow a measured quantity of MgSO4
solution (3.6 per cent, of the dried salt at 1 10° C.) to flow in. Withdraw the
funnel and tie off the opened portion. Replace the loop of intestine and
draw forth another loop; treat this loop also, using i per cent. NaCl instead
of.MgSQ4, and sew up the wound. The NaCl and MgSO4 solutions have the
same freezing-point. Leave until all the other experiments are finished, then
open the abdomen, find the ligated intestines, and measure their contents:
the MgSO4 has not diminished as much as the NaCl, because the former salt
in not readily absorbed and retains water by salt action.
Experiment 2. Saline Diuresis. — Inject warm i per cent. NaCl, 25 c.c.
per kg. (See Exercise I, Experiment i.)
Experiment 3. Strong Vagus Stimulation.
Experiment 4. Theobromin-sodium Salicylate. — Inject 20 mg. (-£• c.c.
of 10 per cent.) per kg. The urine flow increases. Note that the changes
in the carotid pressure do not suffice to explain the diuresis. The effects on
the circulation are identical with those of carotid. The volume of the kid-
ney increases.
Experiment 5. Hemorrhage. — Withdraw about 25 c.c. per kg. of blood
from the femoral artery while taking a tracing. (The blood is to be whipped
vigorously with a glass rod for about ten minutes, or until thoroughly
defibrinated, strained through muslin, and heated to 40° C.)
The ureter flow stops as the pressure falls. The heart-beats are quick-
ened and weakened. The respiration is dyspneic.
The cardiac and respiratory effects are due to anemic depression of the
vagus and respiratory centers. The anuria is explained by the low blood-
pressure.
Observe the pressure for some five minutes after the completion of the
hemorrhage: there is a slight, but very imperfect recovery.
CHAP. XLVI INTESTINAL OSMOSIS-DIURESIS 2Q3
Experiment 6. Injection of Normal Saline Solution. — Urine Flow, Blood-
pressure. — Inject 25 c.c. per kg. of warm normal saline solution. The urine
flow and the blood-pressure recover considerably, but do not usually reach
the original level. The effect lasts for several hours. Note the much larger
effect as compared with saline injection in the normal animal.
Experiment 7. Injection of Defibrinated Blood. — Urine Flow, Blood-
pressure. — After fifteen minutes inject the warmed defibrinated blood: the
ureter flow and blood-pressure recover completely.
SECOND PART: HYDROPERICARDIUM
Observations. — Pulse- rate; blood-pressure tracing; inspection of heart.
Operation. — Start artificial respiration. Expose the heart as in Chapter
XLII, Exercise V. Tie a cannula into the apex of the pericardium. Con-
nect with reservoir of saline.
Experiment 8. Pericardial Pressure. — Study effects of increasing the
pressure by raising the reservoir. Leave this at a level which produces
fairly serious interference with the heart.
Experiment 9. Weak Vagus Stimulation.
Experiment 10. Strong Vagus Stimulation.
Experiment u. Saline Infusion.
Experiment 12. Strophanthus. (See Exercise I, Experiments 5 to 8.)
Experiment 13. When the animal is dead, complete Experiment I.
QUESTIONS
(a) Describe and explain the effects of the procedures of Experiments 2
to 7 on the blood-pressure and ureter flow.
(b) Which of these would be useful in dropsy?
(c) Which in uremia?
(d) Describe and explain the effects of pericardial effusion.
(e) How is this modified by the procedures of Experiments 9 to 12?
(/) Which of these would be useful, which harmful, and which indif-
ferent?
(g) Explain how Epsom salt increases the bulk of the feces.
EXERCISE in.— (GROUP III) FIRST PART: URINE FLOW AND KIDNEY
VOLUME. SECOND PART: MYOCARDITIS
FIRST PART: URINE FLOW AND KIDNEY VOLUME
Observations. — Heart-rate; blood-pressure tracing; ureter flow; kidney
volume; inspection of intestinal vessels.
Apparatus.— Drum; manometer; ureter cannula; oncometer; injection
buret; induction coil.
Operation.— Weigh; etherize; tie on board. Thread under \.
( annulae in carotid, trachea, and femoral vein. Expose kidney and place
in oncometer. Tie ureter cannula in other ureter. Place loop of intestine
for inspection.
Experiment i. Absorption of Sodium Chlorid and Magnesium Sulphate.
— (SeeExerci < II
Experiment 2. Hypertonic Salt Diuresis.— Inject slowly 10 per
NaCl, 2.5 c.c. j>er k^. Compare results with Experiment 2 of Exercise II
The same quantity of \a('l is u>ed, hut the concentration is different.
Experiment 3. Strong Vagus Stimulation.
2Q4 A LABORATORY GUIDE IN PHARMACOLOGY
Experiment 4. Theophyllin-sodium Acetate. — Inject 10 mg. (i c.c. of
1 per cent.) per kg. Results similar to theobromin (see Exercise II, Experi-
ment 4).
SECOND PART: ACUTE MYOCARDITIS
Operation.— Start artificial respiration and expose heart as in Chapter
XLII, Exercise V.
Observations. — Heart-rate; blood-pressure tracing; inspection of heart.
Experiment 5. Injection of Alcohol. — Inject 2 c.c. of 95 per cent. Alcohol
into myocardium. Repeat several times until the blood-preassure has
fallen markedly.
Experiment 6. Weak Vagus Stimulation.
Experiment 7. Strong Vagus Stimulation.
Experiment 8. Saline Infusion.
Experiment 9. Strophanthus.— (See Exercise I, Experiments 5 to 8.)
Experiment 10. When the animal is dead, complete Experiment i.
QUESTIONS
(a) Describe and explain the effects of the procedures of Experiments
2 to 4 on the blood-pressure and ureter flow.
(b) Which of these would be useful in dropsy?
(c) Which in uremia?
(d) Describe and explain the effects of acute myocardial degeneration.
(e) How is this modified by the procedures of Experiments 6 to 9?
(/) Which of these would be useful, which harmful, and which indif-
ferent?
(g) Explain how Epsom salt increases the bulk of the feces.
EXERCISE IV.— (GROUP IV) FIRST PART: URINE FLOW AND KIDNEY
VOLUME. SECOND PART: AORTIC ANEURYSM
FIRST PART: DIURESIS
Observations. — Heart-rate; blood-pressure tracing; ureter flow; kidney
volume; inspection of intestinal vessels.
Apparatus. — Drum; manometer; ureter cannula; oncometer; injection
buret; induction coil.
Operation. — Weigh; etherize; tie on board. Thread under vagus. Can-
nulae in carotid, trachea, and femoral vein. Expose kidney and place in on-
cometer. Tie ureter cannula in other ureter. Place loop of intestine for
inspection.
Experiment i. Glucose Diuresis. — Inject warm 6 per cent, solution,
25 c.c. per kg. (See Exercise I, Experiment i.)
Experiment 2.— Amyl Nitrite. — Administer by inhalation. (See Chapter
XLIII, Exercise I, Experiment 4.)
Experiment 3. Caffein. — Inject 10 mg. (i c.c. of i per cent.) per kg.
(See Exercise II, Experiment 4.)
Experiment 4. Hemorrhage. Experiment 5. Saline. Experiment 6. Re-
mjection of Blood. — See Exercise II, Experiments 5, 6, and 7.
SECOND PART: AORTIC ANEURYSM
Experiment 7. Aortic Aneurysm. — Tie into cardiac end of other carotid
a cannula, the free end of which communicates with a fairly strong but
extensible rubber bulb (made from the finger of a rubber glove, well oiled).
CHAP. XLVI INTESTINAL OSMOSIS-DIURESIS 295
Remove clamp from artery. This simulates a pulsating aneurysm. It
should be watched during the experiment.
Experiment 8. Weak Vagus Stimulation. Experiment 9. Strong Vagus
Stimulation. Experiment 10. Saline Injection. Experiment n. Strophan-
thus. — (See Exercise I, Experiments 5 to 8.)
QUESTIONS
(a) Describe and explain the effects of the procedures of Experiments
i to 6 on the blood-pressure and ureter flow.
(b) Which of these would be useful in dropsy?
(c) Which in uremia?
(d) Describe and explain the effects of aortic aneurysm.
(e) How is this modified by procedures of Experiments 8 to 1 1 ?
(/) Which of these would be useful, which harmful, and which indif-
ferent?
EXERCISE V.— (GROUP V) FIRST PART: URINE FLOW AND KIDNEY VOL-
UME. SECOND PART: CORONARY SCLEROSIS (CARDIOMYOGRAM)
FIRST PART: DIURESIS
Observations. — Heart-rate;, blood-pressure tracing; ureter flow; kidney
volume; inspection of intestinal vessels.
Apparatus. — Drum; manometer; ureter cannula; oncometer; injection
buret; induction coil.
Operations. — Weigh; etherize; tie on board. Thread under vagus.
Cannula? in carotid, trachea, and femoral vein. Expose kidney and place
in oncometer. Tie ureter cannula in other ureter. Place loop of intestine
for inspection.
Experiment i. Absorption of Sodium Chlorid and Magnesium Sulphate.
—(See Exercise II, Experiment i.)
Experiment 2. Saline Diuresis. — Inject, 25 c.c. per kg. of warm Locke
solution (without glucose). (See Exercise I, Experiment i.)
Experiments. Epinephrin.— Inject 0.05 mg. (J$ c.c. of i : 1000) per
kg. (See Chapter XLIII, Exercise I, Experiment 9, p. 271.)
Experiment 4. Pituitary. — Inject solution, o.i c.c. per kg. (See Chapter
XLIII, Exercise II, Experiment 4, p. 273.)
SECOND PART: CORONARY SCLEROSIS
Observation, Apparatus, and Operation for Myocardiogram. — (See
Chapu r XLIV, Exercise I, p. 281.)
Experiment 5. Coronary Sclerosis.— With a hypodermic syringe inject
a suspension of lycopodium1 into a coronary artery.
(Optional) For this may In- substituted:
••//»//;<>«.• Kin I Hi -limul.ition «.|" ;iuri. lc
Kin tri. >tirmil.iti..n "i tin- middlr third of the anterior coronary
arter> .or).
Experiments 6 and 7. Weak and Strong Vagus Stimulation.
Experiment 8. Inhalation of Amyl Nitrite.
Experiment 9. Strophanthus. Inject Strophanthus, as in Chapter
XLIY ; • rimrnt 6, |>. 281.
Experiment 10. \\lu-n the animal is dead, complete Exper inu nt i.
1 Lycopodium Suspauio*.— Lycopodium sports heated and shaken with normal saline, to form
296 A LABORATORY GUIDE IN PHARMACOLOGY
QUESTIONS
(a) Describe and explain the effects of the procedures of Experiments
. 2 to 4 on the blood-pressure and ureter flow.
(b) Which of these would be useful in dropsy?
(c) Which in uremia?
(d) Describe and explain the effects of coronary obstruction.
(e) How is this modified by the procedures of Experiments 6 to 9?
(/) Which of these would be useful, which harmful, and which indif-
ferent?
(g) Explain how Epsom salt increases the bulk of the feces.
EXERCISE VI.— (OPTIONAL) FATE OF INJECTED SALT SOLUTION
Anesthetize a dog. Place cannulae into trachea, carotid artery, femoral vein, both
ureters, and ileum.
Draw a sample (5 or 10 c.c.) of blood, defibrinate, and set aside for the determination
of the ratio of corpuscles and plasma (see Index).
Inject into the vein a 0.9 per cent. NaCl solution, 25 c.c. per kg., in ten minutes. At
the end of the injection draw another sample of blood (defibrinate), and again in half an
hour and in two hours. Collect the urine and the intestinal fluid during the same periods.
Kill the animal and measure the fluid in the intestines, pleura, and peritoneum. Note
whether the liver and lungs are edematous.
Determine the ratio of corpuscles in each of the blood samples. Assuming that the
original volume of blood was 75 c.c. per kg., calculate from these data the distribution of the
injected fluid at each of the periods (Sollmann, 1901, Arch. exp. Path., 46, i).
Sodium sulphate or hypertonic solutions may be used; or gelatin solution, which leaves
the vessels more slowly (1.5 gm. gelatin, melted with 100 c.c. of water, and mixed with 1000
c.c. of 0.9 per cent. NaCl and 2 gm. of sodium carbonate, Hogan, 1915, Jour. Amer. Med.
Assoc., 64, 721).
The Lymph may also be studied (Heinz, 2, 335).
EXERCISE VII.— (OPTIONAL) ANASARCA
The injection of excessive quantities of saline solution into normal animals produces
ascites, but not anasarca (Cohnheim and Lichtheim). True anasarca occurs if saline
solution is injected into an animal poisoned by arsenic (Magnus); or locally, if the
skin is irritated by iodin or hot water.
APPENDIX
APPENDIX A.— ARRANGEMENT AND GENERAL EQUIPMENT OF
LABORATORIES
THE LABORATORY ROOMS
THE pharmacology courses may be given in the chemic, pharmaceutic,
and physiologic laboratories if no other arrangement can be made; but
the efficiency of the teaching and research is undoubtedly enhanced by
separate rooms and equipment. The laboratory should consist of a chemic
and animal department, preferably in adjacent rooms. The materia medica
collection may be placed in the chemical room or in a convenient corridor.
Additional rooms for lectures, research, toxicology, storage, for the keeping
and observation of animals, etc., are highly desirable. They should be in
close vicinity; the animal rooms, however, will be less annoying in another
part of the building.
EQUIPMENT OF THE CHEMICAL DEPARTMENT
This should contain the chemic tables, lockers, and sinks for the students;
a fume-chamber; balance and druggists' scales; and a moderate equipment
of chemic apparatus.
The chemic tables may be of any of the varieties used in chemic labora-
tories. A height of 3 feet is convenient. A working space of 6 by 2 feet
and a single locker suffice for each pair of students. The lockers should
be of the height of the table, 2\ feet wide, with a shelf 9 inches from the top.
It is cheap and convenient to have i-inch iron rods fixed to the tops of the
tables for clamping retort rings, etc.
EQUIPMENT OF THE ANIMAL DEPARTMENT
This should be equipped with a large demonstration table and case of
demonstration apparatus; sinks; easily movable tables and lockers for
students' work; shelves for reagents; a chemic bench; drawers for supplies,
etc.
Tables for Animal Work.— These may be of pine, strongly built, 3 feet
high by 6 feet long and 2 feet wide; ij inch top; solid legs. Drawers are
rather ubjivtinnablr. T\v«> tal>le> are m • -ix students. Inoperative
experiment > tin two tables are set in the form of a T, the lower table beiim
used for operating, the upper one for apparatus.
Black Sttiin for Table Tops.— The clean table is given two coats of the
following solutions: No. i, applied hot, the second as soon as the first is dry.
This is followed by two coats of Solution No. 2 ; this is allowed to dry t '
mmhlv 'one to two days) and sand-papered lightly. It is then paraffined
with floor-wax.
297
298 APPENDIX
Solution No. i :
Copper sulphate i part
Potassium chlorate i part
Water 8 parts
Boil for five minutes.
Solution No. 2 :
Anilin hydrochlorate 3 parts
Water 20 parts
Or,
Anilin (liquid) 6 parts
Hydrochloric acid 9 parts
Water 5° Parts
The lockers (one for six students) may be placed at the side of the room
near the tables. There should also be an open shelf for special apparatus.
Apparatus. — It is advisable to buy as much as possible of manufactured
apparatus of the best quality which the resources will allow. The satis-
faction of working with instruments which give accurate and trustworthy
results, the training in exactness, and the practice with apparatus such as
would actually be used in research are advantages which offset, in most
cases, those of home-made apparatus. The latter, however, have some
valuable qualifications besides cheapness, especially in that they encourage
independence and ingenuity. A certain amount of home-made apparatus is
therefore very useful, especially if tune permits the students to manufacture
it themselves.
APPENDIX B.— EQUIPMENT OF CHEMIC LOCKERS (FOR EACH
PAIR OF STUDENTS)
2 Bunsen burners and tubing. i Mortar and pestle, 10 cm.
1 Retort stand. 2 Keys.
2 Retort rings. i Requisition pad.
1 Tripod. i Percolating tube.
i -liter wash bottle. i Water-bath with rings.
2 Evaporating dishes (10 cm.). i ico-c.c. graduate.
1 Evaporating dish (400 c.c.). i Pint percolator.
2 Funnels, 6 cm. i Pill tile.
1 Funnel, 12 cm. i Pill box.
5 Beakers, 25-150 c.c. i Powder box.
4 Flasks, 250 c.c. i Steel spatula.
2 Tumblers. i Horn spatula.
50 Test-tubes. i Thermometer, 0-100°.
2 Test-tube racks. i 25-c.c. Conic graduate.
2 Test-tube brushes. 3 Watch-glasses, ij inches.
2 Test-tube clamps. i Sponge.
2 Slide clamps. i Towel,
i Earthen jar.
The following are not charged :
Filter-paper; label paper; wire gauze; glass slides, tubing, rods, pipets,
etc.
APPENDIX C REAGENTS NEEDED FOR CHEMIC EXERCISES 2QQ
APPENDIX C.— REAGENTS NEEDED FOR CHEMIC EXERCISES
The reagents employed in pharmacology are so numerous that the
problem of keeping them conveniently accessible is quite serious. It will
be found convenient to divide them into three classes: (A) for every three
students; (B) for every six students; and (C) for every six students for
special experiments. (A) and (B) should be arranged in alphabetic order
on the shelves of the chemic tables. (C) may be arranged by the exercise
numbers, and kept on a side shelf when not in use.
It will be found very advantageous to number the containers and their
places, and to demand that every reagent be replaced in proper order as
soon as used.
A number of the solutions are perishable and should not be kept over
a year. These are marked * in the following lists. Others (**) should be
furnished fresh for each exercise. It is well to distinguish these by colored
labels (green for * and red for **) for the ready guidance of the laboratory
assistant. He can save himself some labor by keeping concentrated stock
solutions on a special shelf.
LIST A.— COMMON CHEMIC REAGENTS
Kept on shelves of chemic tables (50 to 100 c.c. of each). For three
students:
Acid, Acetic, 5 per cent. lodin in KI, i per cent, of iodin,
Acid, Hydrochloric, Cone., C. P. KI q. s. to dissolve
Acid, Hydrochloric, 5 per cent. Lead Acetate, 5 per cent.
Acid, Nitric, Cone., C. P. Litmus Paper.
Acid, Picric, Saturated Aqueous. Magnesia Mixture.1
Acid, Sulphuric, Cone., C. P. Magnesium Sulphate, powdered.
Acid, Sulphuric, 5 per cent. Mercuric Chlorid, i per cent.
Alcohol, Ethyl, 95 per cent. Mercuric-potassic lodid (Mayer's
Ammonia Water, 10 per cent. Reagent).2
Ammonium Sulphate, Powdered. Oleum Olivae or Gossypii (cotton-
Barium Chlorid, 5 per cent. seed).
Barium Hydrate, Saturated Potassic Bichromate, Saturated
Aqueous. (about 3^ per cent.).
**Bromin Water; Saturated Aqueous. Potassic Ferricyanid, 5 per cent.
Calcium Chlorid, i per cent. Potassic Ferrocyanid, 5 per cent.
Calcium Hydrate (Lime Water), Potassic lodid, 3 per cent.
: urated Aqueous. Silver Nitrate, i per cent.
Chloroform. Sodium Acetate, 5 per cent.
Cupric Sulphate, 5 per cent. Sodium Carbonate, 5 per cent.
Ether. Sodium Chlorid, crystal
Ferric Chlorid, o.i per cent. Sodium Hydrate, 10 per cent.
'Ferrous Sulphate, i per cent. Sodium Phosphate, > JUT i rnt.
Glycerin. Sodium Sulphate, powdered.
* (Green Label) should not be kept over a year.
'•<! Label) should be freshly made.
' .l/,jKww.i Mnlur,:
'
I
NHj (10 per cent.) 4
... 8
* Mercuric -polo* tic lodid (Mayer's Rr.i
HgCl, ' 5S|m.
K I
Wat <J. I, I.c
30O APPENDIX
LIST B.— ON TOP SHELF OF CHEMIC TABLES. FOR SIX STUDENTS
("Pd." stands for "powdered"). Approximate amounts (grams or cubic
centimeters) :
Acacia, granulated 20
Acetanilid 20
Acid, Boric, Pd 20
Acid, Phosphotungstic (10 per cent, in 4 per cent. HC1) 10
Acid, Sulphuric- ferric (i : 1000, Ferric Chlorid) 10
Acid, Tartaric, Pd. . . 10
Alum, Pd 10
Antipyrin 10
Bismuth Subcarb 20
Caffein i
Calomel 10
Chloral 5
Creta prepar. (Chalk) , . . 25
Ferric Ammon. Citrate, 5 per cent 25
Ferric Chlorid, Tr 25
Formaldehyd, Liq 20
Fuller's Earth 25
Gasolin 50
Glucose, Pd 20
*Guaiac, Tr 10
Hexamethylenamin 10
*Hydrogen Peroxid 20
lodin, Tr 10
Lead Subacetate Sol 25
Methyl Alcohol 20
Millon's Reagent1 20
Morphin Sulph o.i
Phenol Liq 25
*Phenol, 5 per cent 25
Potas. Arsenite, Liq 20
Potas. Bichromate, Pd " 5
Potas. Bromid, Pd 20
Potas. Chlorate, Pd 10
Potas. Nitrate, Pd 10
Potas. Oxalate, Pd 10
Potas. Permanganate, i per cent 25
Quinin Sulphate, Pd o.i
*Quinin Sulphate, o.i per cent, acidulated, aqueous 25
*Quinin Sulphate, saturated aqueous 25
Resorcin, Pd 0.5
Sand 100
Sod. Acetate, Pd 10
Sod. Benzoate, Pd 10
Sod. Bicarbonate 20
Sod. Borate, Pd 20
Sod. Nitrite, Pd 10
Sod. Salicylate, Pd 10
1 Millon's Reagent: Dissolve i part of metallic mercury in i part by weight of cold fuming
nitric acid, cool, and dilute with 2 parts of distilled water. Decant from the sediment. The solu-
tion contains mercuric and mercurous nitrate.
APPENDIX C REAGENTS NEEDED FOR CHEMIC EXERCISES 301
Sod. Thiosulphate (Hyposulphite), Pd 10
Spir. Nitrous Ether 20
Starch 100
Strychnin Sulphate, Pd o.i
Sugar, Cane, Gran 200
Talc, Purif 50
Tannin, Pd 10
Turmeric Paper
Turpentine Oil 50
LIST C.— SPECIAL REAGENTS, ARRANGED BY EXERCISES
This does not include optional experiments, demonstrations, or special
assignments.
The quantities are for six students.
CHAPTER I
*Nicotin, i per cent 5
*Salicin, i per cent 10
Licorice, Fldext 20
Licorice, Fldext, Acidulated 20
Soap-bark, Tr 10
**Rhubarb Infus., 5 per cent 25
**Cinchona Infus., 5 per cent 25
**Acacia, 10 per cent 25
Rosin, Pd 20
CHAPTER II
Cinnamon Oil, in drop bottles 5
*Quick-lime, in .}-gm. portions 3
Quart bottles 3
Alcohol
Peppermint Oil 5
Peppermint Herb, in o.i-gm. portions 3
Digitalis, in i.5-gm. portions 3
*Cinnamon Water 100
Arnica, in lo-gm. portions 3
Cinchona, Pd., in 2o-gm. portions 3
Cod-liver Oil 30
Syrup. 25
Glycyrrhiza, Pd., in 2-gm. portions 6
Excipient, or Glycerite Acacia.
Powder Papers. 60
Capsules, No. 3 60
Zinc Oxid, Pd., in j-^m. portion- 6
Bcnzoinated Lard, in lo-jrm. purti<ui>
Flaxseed, Ground 300
CHAP! i K in
*Strychnin Sulphate, i per i rut . 10
Spir. Ammoii. Anrni 25
25
*Acaciae, Mucil
Sod. Chlorid, Sat. Sol.. 25
302 APPENDIX
CHAPTER V
**Strychnin Sulphate, i : 50,000 30
**Sod. lodate, i per cent 10
**Starch Paste, 2 per cent 25
**Marquis Reagent 10
Ammonium Molybdate i
Morphin Tablets, f grain 12
Opium, Tr 10
Apomorphin Hydrochlorid .' o. i
Apomorphin Hydrochlorid, i : 500 10
Atropin o.i
**Epinephrin, i : 50,000 10
**Acoriite, i : 300 30
Veratrin o.i
CHAPTER VII
**Calx Chlorinata, 5 per cent 10
CHAPTER Vin
**Formaldehyd, i : 50,000 25
*Jorissen Phloroglucin Reagent 15
*Phenylhydrazin HC1, 0.5 per cent 10
*Sod. Nitroprussid, 5 per cent 10
**Milk 50
**Milk with Formaldehyd, o.i c.c. of the Solution per liter 50
*HCN, i : 1000 5
CHAPTER XIII
*Sugar, i per cent 50
Sod. Saccharin, o.i per cent • 50
Glycerin, 10 per cent 50
*Lactose, 10 per cent 50
*Glucose, 10 per cent 50
*Levulose, 10 per cent 50
Magnes, Sulph., 20 per cent 100
KBr, 5 per cent 100
Sod. Salicylate, 10 per cent 100
Chloral, 10 per cent 100
Quinin Bisulph., i per cent 100
Each of the following 25 c.c.:
Magnes. Sulph., 2% in water; in milk; in 5% acacia; in syrup.
KBr, 0.5% in water;
Sod. Salicyl., i% in water;
Ammon. Chlorid, 0.5% in water; "
Chloral, i% in water;
Quinin Bisulph., 0.1% in water;
Also above list in Syr. Citric Acid; Syr. Glycyrrhizae; Elixir;
Comp.Tr. Cardamom; Syr. Eriodictyon.
Saccharin, o.i per cent 25
Saccharin, o.oi per cent 25
Cod-liver Oil 25
Cod-liver Oil with 0.4 per cent. Oil Peppermint -. . 25
Cod-liver Oil with 0.4 per cent. Oil Lemon 25
a « « «
« « «
u t< «
it « «
-
APPENDIX C REAGENTS NEEDED FOR CHEMIC EXERCISES 303
L
of
10
Toronto
Cod-liver Oil, 50 per cent. Emulsion, not flavored,
Chalk
Chalk, 5; Milk-sugar, 5
Chalk, 5; Cane-sugar, 5
Chalk, 5 ; Cane-sugar, 3 ; Cacao, 2
Chalk, 5; Cane-sugar, 4; Cinnamon, i.
Euquinin (Quinin-ethyl Carbonate)
Quinin-Fuller's Earth Precipitate
Quinin Tannate
Quinin Alkaloid
Quinin Sulphate
Magnes. Sulph., 5 per cent 25
Sod. Sulph., 5 per cent 25
Sod. Phosphate, 5 per cent 25
Sod. Pot. Tartrate, 5 per cent 25
CHAPTER XV
Weighed drugs for each exercise.
CHAPTER XVI
*Strychnin Sulph., o.i per cent 10
*Morphin Sulph., o.i per cent 10
**Infusion Tea, 5 per cent 25
'"Infusion Coffee, 5 per cent 25
**Egg-white Solution, 1:5 25
Phosphorus, 10 small pieces, shot size.
CHAPTER XVII
25 c.c. of each of the following, in wide-mouth jars:
*Citric acid, i per cent, in water.
*Quassia, -f$ per cent, in water.
Ouinin Bisulph., ^ per cent, in water.
*Sugar, 5 per cent, in water.
*Ditto in 10 per cent, starch
paste.
Salt, 3 per cent, in water.
Methylene-blue, 1.5 : 1000 20
Animal charcoal o.i
CHAPTER XXII
Defibrinated blood 25
Cl, 0.9 per cent 15
*NaCl, 0.9 per cent. + -^ Per cent, saponin 15
*NaCl, 0.9 per cent. -j- -^ Per cent- saponin, 15 c.c., digested with 6
drops of i per cent, cholesterin.
*NaCI, 0.9 per cent., sat urutrd with rtlu-r . 15
*Urea, i per cent
Sod. Carbonate, 2 per cent 15
(II AFTER XXV
**Egg-white, i : 100 c.c. water.
ubrinaU-d blood.. 50
**M;mim;ili;r -kin.
'1C.
muscle.
304
APPENDIX
CHAPTER XXVI
Soap-bark 10
*Aconite, i per cent 10
**Egg-white i
CHAPTF.R XXIX
**Milk 50
Rennin i
*t*Barley Decoction, 10 per cent. . . 10
**Pancreatin, o.i per cent 10
*Formaldehyd, o.i per cent 10
*Sod. Citrate, i per cent 10
APPENDIX D.— CONTENTS OF LOCKERS FOR PHARMACODY-
NAMIC EXERCISES
Top Shelf
3 semicircular stands, 5 clamps.
2 induction coils, 2 electrodes.
i ether mask.
Second Shelf
i perfusion bottle, i Woulff bottle.
1 funnel, 2 flasks — 250 c.c.
2 tumblers, 2 beakers.
2 electric keys, i oncometer and
clamp.
2 evaporating dishes, 2 frog boards,
i mesentery board, i foot board,
i dissecting needle, parchment, wax
slides,
sandpaper, two 25 c.c. graduates.
Third Shelf
1 aneurysm needle, two 10 gm. lead
weights.
4 Mohr clamps, 3 bulldogs.
2 hemostats, i electric connection,
i cork with pins, i knitting needle,
i brass T, 2 pithing wires.
1 box with 2 glass Y, with i glass T,
and 5 vessel cannulae.
2 camel's hair brushes.
2 tracheal cannulae, i large screw-
clamp.
1 tracheal tube, i small screw-clamp.
2 heart levers.
4 muscle levers (2 straight, 2 elbow).
4 watch-glasses.
2 bundles ligatures, i suture needle.
2 feathers.
Wall
two 10 c.c. pipets in I25-.
two 10 c.c. pipets in ^.
i clinical thermometer,
i thermometer, i° to 100° C.
i blood-pressure pipet.
i electric signal marker,
i stirring rod.
i syringe, 10 c.c.
i syringe, i c.c.
4 needles in bottle.
1 femur clamp.
Bottom Shelf
2 Harvard kymographs and vane.
2 drums; one loo-c.c. cylindric grad-
uate,
i stomach bulb and tube.
1 small gag, i large gag.
2 clamps (G).
1 mercury manometer and guide.
Drawer, Left
2 towels, i sponge, i set ropes.
Cupboard
i saucepan, 10 test-tubes with rack1
and brush.
Wooden bench.
Bottles of saline (0.75 and 0.9 per
cent.), ether, and MgSO4-
Artificial respiration bellows,
i buret (25 c.c.), stand, clamp, and
tube.
APPENDIX E SOLUTIONS FOR PHARMACODYNAMIC EXERCISES
305
APPENDIX E.
No. of
bottles.
2
2
2
2
6
2
2
I
2
2
I
2
2
3
2
2
2
2
2
3
2
2
2
3
2
4
4
2
3
i
2
3
2
2
2
2
2
2
4
2
2
2
2
I
2
6
2
3
ALPHABETIC LIST OF SOLUTIONS NEEDED FOR
PHARMACODYNAMIC EXERCISES
I)rui;s.
Strength.
25 per cent.
. 5 per cent.
. 5 per cent.
0.5 per cent.
**Acacia
Acid, Acetic
Acid Fuchsin
Acid, Hydrochloric
**Acid, Hydrochloric, 0.5 per cent, in 10 per cent. Acacia
*Acid, Hydrocyanic 2 per cent.
*Acid, Lactic 0.6 per cent.
Acid, Nitric Cone.
Aconite (Tinct.) 10 per cent.
**Aconite 4 per cent.
**Aconitin i : 10,000
Alcohol 95 per cent.
Alcohol 50 per cent.
Alcohol 25 per cent.
Alcohol 10 per cent, in N. S.
Alcohol i : loo in N. S.
Alcohol i : 1000 in N. S.
*Alypin i per cent.
Ammon. Chlorid i per cent.
Ammonia Water
Amyl Nitrite
Antimonium Potas. Tartrate J per cent.
*Antipyrin 2 per cent.
**Apomorphin i per cent.
**Apomorphin i : 1000 N. S.
*Atropin Sulphate i per cent.
*Atropin Sulphate i : 1000
Barium Chlorid i : 1000
Barium Chlorid i per cent, in N. S.
*Beta-tetra-liydro-napthylamin 5 per cent.
** Bismuth Suspension
*Caffein i : 100
*Caffein i : 1000
*Caffein. . i : 5000
*Caffein i : 10,000
Calcium Chlorid 10 per cent.
Calcium Chlorid S per cent.
Calcium Chlorid 1.6 per cent.
Calcium Chlorid. . . . i per cent, in N. J
Calcium Chlorid., 0.15 per cent, in O.Q per cent. NaCl
*Camph<>r .. 20 per cent, in oil
miplmr i prr cent, in 40 per cent, alcohol
'.mphur SafdinN. S.
icum IVtrolatum
Mir]
*(Y\adin. i : 1000
Chloral HydraU- . 25 per cent.
Chloral H\drau- 10 per cent.
Sire.
( 30 c.c.)
(200 c.c.)
( 5 c.c.)
( 10 C.C.)
( 10 C.C.)
( 5c.c.)
( 20 C.C.)
( 50 C.C.)
( 15 C.C.)
( 5c.c.)
( i c.c.)
( IS c.c.)
( 5 c.c.)
( 50 c.c.)
( 25 c.c.)
( 25 c.c.)
( 25 c.c.)
( ic.c.)
(150 c.c.)
( 10 C.C.)
( 5c.c.)
( 50 c.c.)
( 25 c.c.)
( 10 C.C.)
( 5c.c.)
( 10 C.C.)
( 10 C.C.)
( 5 c.c.)
( 25 c.c.)
( 5 c.c.)
( 50 c.c.)
( 25 c.c.)
( 25 c.c.)
( 5 c.c.)
( 25 c.c.)
( Sc.c.)
( 10 C.C.)
(200 c.c.)
( 25 c.c.)
(200 c.c.)
( 5°
( 25 c.c.)
( 10 C.C.)
( ic.c.)
( 10
( 10 C.C.)
( logm.)
(100
( 25
300 APPENDIX
No. of
bottles. Drugs. Strength. Size.-
2 Chloral Hydrate 2 per cent. ( 5 c.c.)
2 Chloral Hydrate i per cent. ( 5 c.c.)
2 "Chloroform Sat'd in N. S. ( 20 c.c.)
6 "Chloroform (50 c.c.}
1 **Cholin i : 1000 ( 20 c.c.)
2 **Cocain Hydrochlorid 5 per cent. ( 2 c.c.)
2 **Cocain Hydrochlorid 2 per cent. ( 5 c.c.)
6 **Cocain Hydrochlorid i per cent. ( 2 c.c.)
2 Colchicum, Fid. Ext ( 5 c.c.}
2 Copper Sulphate i per cent. ( 50 c.c.)
1 **Cotarnin i : 1000 ( 10 c.c.)
3 *Curare \ per cent. ( 15 c.c.)
2 *Curare i : 1000 N. S. ( 5 c.c.)
3 Digitalis (Tinct.) 10 per cent. ( 10 c.c.)
2 **Digitalis 4 per cent. (100 c.c.)
2 **Digitalis i per cent. ( 10 c.c.)
2 *Dionin 10 per cent. ( i c.c.)
6 Epinephrin i mg. tablets (i tube)
2 Ergot, Fid. Ext ( 5 c.c.)
2 Ergot (Tinct.) 10 per cent. ( 10 c.c.)
2 *Ether Sat'd in N. S. ( 20 c.c.)
i *Fluorescein Sol ( 5 c.c.)
i **Glucose 6 per cent. (400 c.c.)
i Histamin Tablets ( i mg.)
1 *Hydrastinin 2 per cent. ( 10 c.c.)
2 Hydrastis (Tinct.) 10 per cent. ( 5 c.c.)
2 **Hydrastis 2 per cent. ( 10 c.c.)
i Lead Acetate Paper.
1 *Lycopodium Suspension ( 10 c.c.)
2 Magnesium Chlorid 2.1 per cent. (200 c.c.)
2 Magnesium Sulphate " 25 per cent. ( 10 c.c.)
2 Magnesium Sulphate 5 per cent. ( 5 c.c.)
4 Magnesium Sulphate 3.6 per cent, (dried) ( 25 c.c.)
2 Mercuric Chlorid. -. i : 1000 (15 c.c.)
2 *Morphin Hydrochlorid or Sulphate 4 per cent. ( 25 c.c.)
2 *Morphin Hydrochlorid i : 1000 ( 5 c.c.)
i **Muscarin o.i per cent. ( i c.c.)
1 Nicotin Undil. ( i c.c.)
2 *Nicotin i per cent. ( 5 c.c.)
2 *Nicotin i : 1000 N. S. ( 5 c.c!)
2 Nitroglycerin i : 100 ( 5 c.c.)
4 *Xitroglycerin i : 1000 ( 10 c.c.)
2 "Novocain i per cent. ( i c.c.)
2 *Ouabain . i : 1000 ( i c.c.)
2 **Ouabain i : 10,000 ( 2 c.c.)
3 **Ouabain i : 50,000 ( 2 c.c.)
2 **Peptone, Witte's 20 per cent. ( 25 c.c.)
2 "Phenol i per cent. (100 c.c.)
3 **Physostigmin Salicylate or Sulphate i per cent. ( i c.c.)
4 **Physostigmin Salicylate or Sulphate. . . . i : 1000 N. S. ( 5 c.c.)
2 *Picrotoxin '. i : 250 ( 5 c.c.)'
APPENDIX E SOLUTIONS FOR PHARMACODYNAMIC EXERCISES
307
No. of
bottles.
3
2
Drugs.
*Pilocarpin Hydrochl
*Pilocarpin Hvdrochl
Strength.
i per cent.
i : looo
Sire.
( sc.c.)
( IS C.C.)
4
2
2
2
2
Potassium Chlorid
Potassium Chlorid
Potassium Chlorid
Potassium Chlorid
Potassium Chlorid
. . . . 10 per cent.
i per cent.
. . . . i : 100 N. S.
. .. i : 1000 N. S.
. i : 10,000 N. S.
( 10 C.C.)
( 50 c.c.)
( 25 c.c.)
( 25 c.c.)
( 2C. c.c.)
2
2
Potassium Permanganate
Quinin Hvdrochlorid
i per cent,
i : loo N S.
( 5° c.c.)
( 2S C C )
2
3
2
Ouinin Hvdrochlorid
*Ouinin Hvdrochlorid
**Quinin-urea Hvdrochl.
. . i : 1000 N. S.
. i : 10,000 N. S.
i per cent.
( 25 c.c.)
( 25 c.c.)
( I C.C.)
6
Ringer's Solution
(dOO C C )
2
2
Ringer's Solution
Ringer's Solution.
without Ca
without K
( 10 C.C.)
( IO C C.)
2
Ringer's Solution. with
CaClj 0.8 • looo
( IO C C.)
2
Ringer's Solution
. Triple strength
( IO C.C.)
2
*Saponin
i : looo N. S.
( 5 c.c.)
2
*Scopolamin Hydrobromid.
I ' IOOO
( 2 C C.)
2
Silver Nitrate
i per cent.
( «; c.c.)
2
Sod. Acid Phosphate.
2 per cent.
(200 c.c.)
i
Sodium Arsenate
5 per cent.
( IO C.C.)
2
Sodium Chlorid
. . Powder
( i em.)
2
Sodium Chlorid . .
10 per cent.
( co c.c.)
2
Sodium Chlorid
c ner cent.
(CQO C C.)
2
4
2
Sodium Chlorid
*Sodium Citrate
Sodium Citrate
2 per cent.
5 per cent.
... 2.7 per cent.
(200 c.c.)
( 25
Uoo c.c.)
2
2
Sodium Fluorid
*Sodium Nitrite
... 0.5 per cent.
. 10 per cent.
( 25
( ^
2
2
*Sc>(lium Nitrite
*Sodium Nitrite
i per cent.
... o.i per cent.
( 5
( s;
2
Sodium Phosphate
. . 2.1 per cent.
(200 c.c.)
I
*Sodium Santonin
5 per cent.
( 2^ C.C.)
2
2
Sodium Sulphate
Sodium Sulphate
2.5 per cent.
i.() per cent.
(400
(200 c.c.)
I
2
2
Sodium Verona 1
•<•;:.
*Siuvain
. . . 10 per cent.
i pi-r iTiit.
i per cent.
( 25
( 15 c.c.)
( I
2
C
phanthufl
phanthus.
i : 100
. i : 1000
( sc.c.)
( 1C
2
6
2
2
2
2
2
2
2
2
ilphau- or Nitrate)
• linin (Sulphate or Nitrate)
*Strychnin (Sulphate or Nitrate
**Sugar, Cane.
"Tannin,
**Tannin
*Theobrom. Sod. Salic, or A« «-i
Theobrom. Sod Salic, or Acet
>l<mm. Sod. Salic, or Acet
i : 100
i : looo
i : 10,000
i pel i-ent.
10 per cent.
I JUT iTIlt.
10 per cent.
i : ioo
i : looo
i : 10,000
( 5
( 20
( 5
(20C
( 5
( Sc.c.)
( 5
( 25
( 25 c.c.)
308 APPENDIX
No. of
bottles. Drugs. Strength. Size.
1 *Theophyllin Sod. Acetate 10 per cent. ( 10 c.c.)
2 *Tropacocain i per cent. ( i c.c.)
1 "Tyramin i : 1000 ( 20 c.c.)
2 **Urea 1.9 per cent. (200 c.c.)
6 *Urethane. . . / 10 per cent. ( 2 c.c.)
2 *Veratrin i : 10,000 ( i c.c.)
6 **Veratrin i : 1,000,000 ( 25 c.c.)
2 Veratrum (Tinct.) 10 per cent. ( 5 c.c.)
4 Zinc Sulphate i per cent. ( 50 c.c.)
TWO (2) LITER BOTTLES
Locke's Solution — Glucose free (2 bottles).
Ringer's Solution.
Sodium Chlorid, 10 per cent.; 2 per cent.; i per cent. (5 bottles); o.o per
cent.; 0.75 per cent.
Tyrode's Solution (Glucose free).
APPENDIX F
TABULATION OF ANIMALS
309
1
§
1
sr
!
a.
£££M«OO<»^» O»cn .* U) tt »H
1
(M to to
(A (A C»> CM
£*t6t t $ s <& s !!!!-
<&<&££
f
oonnnnonoooononooooonooo
D= Demonstration.
C- Class.
( ' : •* : : •<> : ^ :
Total.
y?
...
• « o . ->» :
New.
t^ j
Sl
X i
o « • ^j •
Survive.
• • •
(^tntatntn; K>; ; ; «; « ; M M M N ; ; ; ; ; ; ;
Total. g,|
.... e ••
Cn(/icn(yi] o! ! ! O! **! Mto^wi | ! ! '. '. \
N... *|
'. '. '. oo 3 '. ON o^cn *>;;;;;;;!;;
Total.
5, :*.o:ooo«^*-::::::::::
v
::;^4*.:tn4».cn*.....:
Survive.
K> c*j K> K> o^c*» 4»- ; >-« ; ; ; *« *^ *» *V •
Total.
9 o-^oo>o«^ io::;---©::-1
New. |
K> u> M o 4^ o ^ ! o; ; ; "-ooo; ; •->
Survive.
2 !!tnK>tn;;!!!;;
Turtles.
§o r ' -vi ^»t/i • o o o M ^
Ordinary. •«
<*» ! ! c*» ! "-1 ! ! ! M
Lui*.
^ <- »
0
- • c 1 . . .
i f "
I
: sifi
i **$*
1
f
3io
APPENDIX
APPENDIX G.— SOLUTIONS AND MATERIALS NEEDED FOR
INDIVIDUAL PHARMACODYNAMIC EXERCISES
CHAPTER XXXII.— LOCATIONS OF ACTIONS, ETC.
Groups or •
Demonstration. Exercise. Animals.
Demonstrations: 1 7 Frogs.
II Frog.
IV 5 Frogs.
V Rabbit.
Frog.
All A Groups:
I Frog.
Ill Frog.
V
Groups: I, A
Ill
V
II, A
III
V
III, A
III
V
IV, A
Ill
* » f *»
V
V,A
III
V 2 Frogs.
All B Groups:
II Frog.
V Frog.
I, B
II
y
'IV . .
V
II, B
i i . u
II
IV
V
m, B
II
IV
Solutions.1 '
Strychnin, i : 1000 (J).
Acetic Acid, 5% (10).
Acid Fuchsin, 5% (2).
Picrotoxin, i : 250 (1.5).
Veratrin, i : 10,000 (£).
Caffein, i : 100 (i).
0.75% NaCl (200).
Ether (10).
Curare, \% (2).
Nicotin, i : 1000 (i).
Tobacco (5).
Cocain, i% (£).
Quinin-urea HC1, i% Q).
Tr. Aconite (10).
Ethyl Chlorid (10).
Strychnin, i : 1000 (}).
HC1, 0.5% do).
Cocain, i% (2).
HC1, 0.5% in 15% Acacia
do).
HCN, 2% (i).
Alcohol, 10% (*).
Stovain, i% (i).
Urethane, 10% (2).
Quinin-urea, HC1 i% (i).
Morphin, 4% (£).
Magn. Sulph., 25% (i).
Strychnin, i : 10,000 (£).
Epinephrin, i : 1000 (i).2
Epinephrin, 0.1% with Co-
cain, i% (i).
(HC1, 0.5%, use that of A
Groups.)
(Cocain, i%, use that of A
Groups.)
Morphin, 4% (2).
Curare, i : 1000 N. S. (5).
Physostigmin, i : 1000 N.
S. (3)-
Novocain, i% (i).
Alcohol, 25% (2).
Nicotin, i : 1000 N. S. (5).
Physostigmin, i : 1000 N.
S. (3);
Novocain, i% (i).
Chloral, 2% (i).
Magnes. Sulph., 5% (5).
Physostigmin. i : 1000 N.
Special apparatus.
Pipet and needle.
Tenaculum.
Oil-bath.
Strong scissors.
Fine scissors.
Fine forceps.
Fine ligatures.
Inductorium (single
shocks).
Aortic cannula.
Perfusion bottle.
Small cannula.
Bell-jar.
Cotton.
Bell-jar and aspira-
tor.
Mounted bristles.
Basin.
Quinin-urea HCl, i% (i).
1 The figures in parentheses are the cubic (.entimeters used in the experiment.
2 Tablets.
APPENDIX G
INDIVIDUAL PHARMACODVNAMIC EXERCISES
Groups or
Demonstration.
All B Groups:
IV, B
Exercise. AnimaU.
Solutions.
IV Saix>nin, i : 1000 N. S. (3).
\' Tn.paoKain. i' 'c (i).
\. I; II . tiagnes. Sulph., .•;' (i).
Apomorphin, i : 1000 N. S. (3).
Alypin. i%(i).
Total Animals Needed. — Demonstrations: 14 frogs, i rabbit (s).1 Class Work: 22
frogs.
CHAPTER XXXm.— MUSCULAR CONTRACTIONS
Groups or
Demonstration. Exercise. Animals.
Demonstrations: III 2 Frogs.
IV 3 Frogs.
A II Groups:
\ <>r B;.
CroU;, I
(inmp II:
Croup ill
II
VII
II
MI
Solutions.
Caffein, i : 10,000 X >
Caffein, i : 1000 X. S. (25).
Ouinin HC'l. i : 10.000 X.
S. (25).
Quinin HC1, i : 1000 X. S,
(25).
Alcohol, i : 100 N. S. (25).
VI Large Frog.
VIII 3 Frogs. NaCl, 10% (5).
IX
X 2 Frogs.
I Frog.
I
VII
I
Ringer's* Solution (5).
Ringer's Solution without
Ca (5).
Ringer's Solution without
K(5).
Ringer's Solution, triple
strength (5).
Ether, sat'd in N. S. (10).
Tannin, i% (5).
/.in. Sulphate, i% (5).
Copper Sulphate, ic'c (5).
Tannin, 10% (5).
Kpincphrin, i : 1000 (5).
Veratrin, i : 10,000 (i).
Caffein, i : 10,000, i : 1000,
i : 100 N.S. (
CaCU, 1% in N. S. (25).
Sod. Citrate, 5% (25).
Theobromin Sod. Salir..
i : 10,000. i : looo. I : IOO
N. S. (25).
Sod. Citr.
Barium Chl.irid. i' , in N.
S. (25).
Uuinin IK 'I. i : 10,000,
i : looo. i : 100 \
(25).
KCI. .',
im Chlorid, i% («).
B;iri»imC'hl..ri.l. .' , in N.S.
(s; — survives.
Special apparatus.
Muscle lever, induc-
tion coil, and ky-
mograph, set up
for tracing as pat-
tern.
Maximal load spring.
Arrangement for Kx-
ercise IV.
Perfusion bottle set
up, with water.
Aorta cannula.
Stand and hook.
Operating instru-
ments.
XI. i, ne I.xperi-
ment.
Lung arranged as
pattern.
APPENDIX
Groups or
Demonstration.
Group IV:
Exercise. Animals.
Solutions.
I KCI, i : 10,000, i : 1000,
i :iooN. S. (25).
VII Sod. Citrate, 5% (25).
KCI, 0.1% in N. S. (25).
Group V: I Alcohol, i : 1000, i : 100,
i : 10 N. S. (25).
VII Sod. Fluorid, 0.5% (25).
Calcium Chlorid, i% in N.
S. (25).
Total Animals Needed. — Demonstration: i large frog, 10 ordinary frogs.
Work: 10 frogs (half class).
Class
Groups or
Demonstration.
Demonstrations:
CHAPTER XXXIV.— SMOOTH MUSCLE
Exercise. Animals. Solutions.
All Groups:
Group I:
Group II:
Group III:
Group IV:
I Decere-
brated
Rabbit.
V Female
Rabbit.
Sheep's
Carotid.
VI to
IX
VI,
VIII,
IX
VII
VI
Viand
VIII
VII
IX
VI
VII
VIII
IX
VI
VII
VIII
IX
NaCl (*).
Physostigmin,
(i).
Barium Chlorid, i% (4).
Atropin, &% (4).
Nicotin, i% (i).
Normal saline (200).
Pilocarpin, &% (15).
Pituitary Solution (1.5).
Warm Tyrode Solution
(2000).
Warm Normal Saline, 0.9
(3000).
Oxygen.
2 Cylinders of Tyrode Solu-
tion.
Sod. Nitrite, 10% (i).
Epinephrin, i : 10,000 (J).
Pilocarpin, i : 1000 (i).
Atropin, i : 1000 (i).
Barium Chlorid, 10% (2).
Sod. Sulphate, 1.9% (200).
Atropin, i% (i).
Pituitary Solution, (£).
Sod. Citrate, 2.7% (200).
Epinephrin, i : 10,000 (J).
Pilocarpin, i% (i).
Atropin, i% (i).
Magnesium Chlorid, 2.1%
(200).
Quinin HC1, i% (i).
Barium Chlorid, i% (5).
Nicotin, i% (i).
Atropin, i% (i).
Calc. Chlorid, 0.15% in 0.9
NaCl (200).
F. E. Ergot (i).
Tr. Digitalis (i).
Special apparatus.
Operating instru-
ments.
Board.
Injection buret,
connections, and
clamp.
Hypodermic syringe.
Tracheal cannula.
Vein cannula.
Respiration bellows.
Bell-jar.
CO« apparatus.
Induction coil.
Ligatures.
Water-bath at 40° C.
Aortic cannula.
for
Dish and rods
defibrinating.
Lever, etc., set up as
pattern.
Water-bath at 40° C
Air current.
APPENDIX G
INDIVIDUAL PHARMACOD\NAMIC EXERCISES
313
Groups or
Demonstration. Exercise. Animals. Solutions.
Group V: VI . Barium Chlorid, i' 'c (5).
Atropin, i' , (i).
VII XaCl. :', ;oo).
Mil Tr. Hydrastis (i).
IX Physostigmin, i% (i).
Total Animals X ceded.— Demonstrations: Decerebrated rabbit (f).1 Class Work:
Female rabbit (f) (Half class).
Groups or
Demonstration.
•nslralions:
CHAPTER XXXV.— PERFUSION EXPERIMENTS
Exercise. Animals.
I White-
Rabbit.
II Rooster.
IV Large Frog.
Solutions.
Xicotin, 1% (i).
F. E. Ergot (5).
Sod. Nitrite, 0.1% (i).
VII
2 Morphin-
ized dogs.
All Groups:
Group I:
Group II:
(irmip HI
p IV:
Group V:
Groups n.m
l\ \
vn
IX
vii
VII
VII
vn
\n
IX
I\
Epinephrin, i : 5,000,000(1).
Digitalis, i : 100 (i).
Ringer's Fid. (500).
Ether (100).
Norm. Saline (2000).
NaCl, i% (1000).
cept Group I.)
(Ex-
Amyl Nitrite (i).
!, 2% (2000).
NaCl, 5% (500).
Calc. Chlorid, 1.6% (200).
Sod. Citrate, 2.75% (500).
Kpim-phrin, i : 1000 (i).
HCN, 2% (2).
Tr. Digitalis (i).
Chloral. 10% (i).
Barium « lilorid. i% (5).
Drl'ihriiKitrd blood (200).
II* V ', (2).
(2).
Tr Digitalis (i).
i o.ooo (5).
\itrilc. i : 100 (5).
li-. i : 100 (5).
Chloral
Barium (hl<. rid. i : 1000(5).
Special apparatus.
Hypodermic syringe.
Ma riot te bottle with
connection and
clamp, on stand.
Forceps, coarse and
fine.
Strong and fine scis-
sors.
Fine ligatures.
Frog board.
Drop-counter.
Aortic cannula.
Vein cannula.
Perfusion bulb.
Operating instru-
ments.
5 cannulae (renal ar-
tery); carotid and
femoral cannula.
Dish, rods, and
strainer for
blood.
Perfusion stand with
connections.
Oncometer bulbs.
ictcr.
I >• n Whitr ral)l>it (s); Rooster (s); Large frog.
Class -s).
' Fatal.
APPENDIX
Groups or
Demonstration.
Demonstrations:
CHAPTER XXXVI. -EXCISED HEARTS
Exercise. Animals.
I Morphin-
ized dog.
Solutions.
Kllier, 100.
Warm Locke Fluid, 3000.
( Kygen.
Strychnin, i : 5000 (5).
Caffein, i : 5000 (5).
Chloroform, sat'd in N. S.
(5)-
Epinephrin, i : 10,000 (5).
KC1, i : 100 (5).
Camphor, sat'd in N. S. (5).
Digitalis, i : 100 (5).
II Large Frog. Ringer's Solution (50).
Ca-free Ringer (10).
Ringer with Calcium Chlo-
rid 0.8 : 1000 (10).
Aconitin, i : 10,000 (i).
Potassium Chlorid, 10%
(0.5)-.
Strychnin Sulph., i : 1000
(i).
Strychnin Sulph., i : 100
(i):
Caffein, i : 100 (i).
Epinephrin, i : 10,000 (i).
Ill 6 Frogs of Ouabain, i : 50,000 (1.5).
abt. 20 gm.
Tr. Digitalis (i).
diluted £ (1.2).
V Fr6g.
VI Turtle.
VII Turtle.
VIII Frog.
Class Work:
All groups:
Groups I, II:
Groups III, IV
IV
VI
IV
IV
Frog.
Turtle.
Groups I, II,
III, IV:
Group I:
VI
VI
Group II:
IX
VI
IX
Group III:
VI
IX
Ringer's Solution (2000).
Pilocarpin HC1, 0.5% (i).
Atropin Sulph., 0.1% (i).
Muscarin or physostigmin,
0.1% (i).
Urethane, 10% (2).
Ringer's Solution (250).
Digitalis, Tr. (i).
Aconite, 4% (5).
Epinephrin, i : 100,000 (i).
Antipyrin, i% (2.5).
Alcohol (10).
Tr. Aconite (2).
Strychnin, i : 1000 (2).
Caffein, i : 100 (2).
Alcohol (7).
Ouabain, i : 10,000 (2).
Special apparatus.
Langendorff appa-
ratus.
Injection buret and
funnel.
Operating instru-
ments.
Bone-forceps.
Ligatures.
Cannulae for carotid,
femoral, trachea,
and aorta.
Dish, rods, funnel,
and strainer for
blood.
Hypodermic syringe.
Straub-Fuehner can-
nula,etc.,Ex. II, 2.
Pithing needle.
Pipet and needle.
Heart lever and
stand set up for
pattern.
Perfusion bottle and
cannulae, set up
for pattern.
Hammer.
Bone-forceps.
Saw.
Turtle-lever and
drum.
Induction coil.
APPENDIX G
INDIVIDUAL PHARMACODYNAMIC KXLRCISES
315
Groups or
Demonstration.
Group IV
Group V:
Exercise. AnimaU.
VI. IX
IX
IV
Solutions.
KCI. 10', (7j.
Kpinephrin. o.i( ( (2).
Chloroform in X. S. (10).
10).
VI Tr. Digitalis (o.i).
KC1, 10' t (i).
IX Cak. Chlorid, 10^ (2).
Kpinephrin, o.i' . < (2).
Total Animals Needed. — Demonstrations: Morphinized dog (f); 9 frogs (i large, 2
medium, 6 of about 20 gm.); 2 turtles. Class Work: 5 frogs; 5 turtles (for half class).
Demonstration.
nitrations:
CHAPTER XXXVII.— PUPILS, ETC.
Exercise. Animals. Solutions.
I Morphin- Ether (200).
ized dog. Atropin, i : 1000 (2).
.
Physostigmin, i : 1000 (2).
Ill
Special apparatus.
Board.
Operating instru-
ments.
Induction coil.
Injection syringe.
Vein cannula.
Rubber rings (20).
V
j Cat- or
Pilocarpin, i' f (3).
Hypodermic syringe.
Rabbits.
Atropin. i% (10).
VI
Dilute acetic acid (200).
VII
Rabbit.
Kpincphrin, i : 10,000 (2).
Board.
Tracheal cannula.
Pilocarpin, i : 1000 (2).
Motor bellow s.
Histamin, i : 10,000 (i).
T-piece.
Pleural cannula.
Tambour.
Kymograph.
Jugular cannula.
Pithing rod.
VIII
Guinea-pig.
Tyrode solution (Glucose
Pipet.
free) (500).
Peptone, i% in Tyrode
Pulmonary artery
(250).
cannula.
Perfusion bottles.
connections, and
stand.
X
2 Sensitized
Horse serum (2).
guinea-
Chloroform (20).
pigs.
XII
Calcium
Dionin, 10% (|).
cat.
Xormal cat.
Class Work, A Groups:
II Cat.
II II
III II
I\ II Cat.
\ 1 1 Cat.
'.\'ork, R Groups:
III Fro-:.
Group- I. Ill
Group M
Group III
P.
( iroup \
Atropin. o.i'
Piltuarpin. i
Physostigmin. i
:-pin. i' ,
n, i%(i).
Dionin, 10% (|).
rpin.
'(i).
(i).
(J).
Atmpin, |% (J).
(i).
; 'A).
Fpinrphrin. i : 10.000 (J).
i-phrin. i : 100
V ceded.— I ' '-bit (0; 2 cats cr
rabbits (s); 3 guinea-pigs (f); 2 cats (s). Class Work: 5 cats (s); 6 frogs.
APPENDIX
CHAPTER XXXVIII.— ABSORPTION, ETC.
Groups or
Demonstration.
Exercise. Animals.
Solutions.
Special apparatus.
Demonstrations
I Morphin-
Ether (200).
Mercury manometer
ized dog.
and connections.
Epinephrin, i : 1000 (5).
Double kymograph.
Strychnin, i : 100 (3).
Carotid and tracheal
cannulae.
2 Femoral cannulae.
Hypodermic syringe.
Injection buret.
II 2 Rabbits
Nirotin (i).
Pipet.
or cats
HCN, 2% (5).
(sick).
Ill Rat, cat, or
Bell-jar with coal-
guinea-pig.
gas.
VII Rabbit or
Lead acetate paper.
H2S apparatus.
cat.
Rectal tube.
VIII Rabbit with
Fluorescein Solution (i).
iodid, mor-
phin, and
calomel.
Rabbit with
calomel.
Class Work:
Group I
IV 2 Rabbits.
Strychnin, i : 1000 (5).
XVI Cat or dog.
Copper Sulph., i% (50).
XVII Dog.
Morphin, 4% (3).
Group II
V 2 Cats.
Apomorphin, i% (i).
Chloral, 10% (15).
XVI Cat or dog.
Zinc. Sulph., i% (50).
XVII Cat or dog.
Morphin, 4% (3).
Zinc Sulphate, i% (50).
Group III
VI 2 Cats.
Strychnin, i : 1000 (6).
Acacia, 25% (30).
XVIII Cat.
Bismuth Suspension (50).
Zinc Sulph., i% (25).
XVI Cat or dog.
Antim. Potas. Tart., \%
(40).
Group IV
XI Dog and
Ammonia vapor.
rabbit.
Atropin, T% (10).
Group V
XIV i Dog.
Apomorphin, i% (20).
2 Cats.
i Rabbit.
Total Animals Needed. — Demonstrations: Morphinized dog (f)
2 rabbits or cats (s) ; 2 rabbits (s). Class Work: (s) 3 rabbits; 5 cats;
; 2 rabbits or cats (f ) ;
4 dogs or cats; 3 dogs;
(f): i rabbit; i
cat.
CHAPTER XXXIX.— TEMPERATURE. ETC.
Groups or
Demonstration.
Exercise. Animals.
Solutions.
Special apparatus.
Demonstrations
XI Rabbit.
HC1, i% (300).
Stomach-tube.
Vein cannula.
Buret and stand.
XIII White
Morphin, i : 1000 (£).
Hypodermic syringe.
mouse.
XV Rabbit.
Magn. Sulph., 25% (15).
Operating instru-
ments.
Calc. Chlorid, 3% (10).
XX Uranium
rabbit.
XXIII Morphin-
Ether (200).
Blood-pressure.
ized dog.
Acetic acid, 5% (10).
Pipet.
Nitric acid (50).
Injection buret.
XXIV
Sod. Arsenate. <;% do).
APPENDIX G
INDIVIDUAL PHARMACODYNAMIC EXERCISES
317
Groups or
Demonstration.
Class Work:
Group I
Group II
Group in
Group IV
Exercise. Animals.
4 Cats.
Rabbit,
dog, and
cat.
2 Cats, i
dog, and
i rabbit.
2 Cats, i
rabbit or
cat.
i Rabbit.
Solutions.
Chloral, 2.5% (200).
Caffein, i% (3).
Strychnin, 0.1% (i).
Morphin, 4% d
F. E. Cplchicum (5).
Mercuric Chlor., i : 1000 (15).
Sod. Arsenate, i% (3).
Sod. Santonin. 5' t (25).
Ext. Cannabis (0.5) in cap-
sules.
Alcohol, 25% (25).
Zinc Sulphate, i% (25).
Cocain, 5% (2).
Beta-tet ra-hydro-naph thy 1-
amin, 5% (3).
Alcohol, 25% (25):
Caffein, i% (15).
Group V 3 Rabbits. Witte Peptone, 20% (25).
Antipyrin, 2% (25).
Total Animals Needed. — Demonstrations: Morphinized dog (f); rabbits, 3 (f); white
mouse (s). Class Work: (f) 15 cats; 2 rabbits; i dog; i rabbit or cat; (s) 4 cats; 4 rabbits;
i dog.
CHAPTER XL.— CONVULSANTS, ETC.
Groups or
Demonstration.
Class Work:
Group I
Exercise. Animals.
Solutions.
Camphor, 20^ in oil (60).
Chloroform (10).
Groups II to V
Group III
Group IV
Group V
i Cat or
rabbit.
1 Bromid
cat or
rabbit.
2 Cats each. Strychnin, i : 1000 (5).
Chloral, 2.5% (30).
Potas. Permanganate, i% (50).
Charcoal (25).
Hydrocyanic acid, i : 1000 (10).
Potas. Permang., i% (50).
Total Animals Needed.— Class Work: 2 rabbits or cats (f); 4 cats (f); 4 cats (s).
Demonstrations:
Groups I II,
and 1 1 1
Group I
Group II
III
Group* IV, \
CHAPTER XLI.— RESPIRATION
Exercise. Animals. Solutions.
I 2 RahhiN. Morphin. i : 1000 (3).
M< •• ; 'i ;•:. :' , (3).
Camphor. JQ' , in oil (1.5).
Caffein, i% (
Rabbit.
IV
VI
Ml.
MM
' , (50).
um Petrolatum (i).
Chloral
in,
nin. o.i' \ (3).
Atropin.
Ammonium Mmvini; l».«t tlr.
Morphin. i : iocx> (2).
Morphi' ;
*)•
Special apparatus.
Trnnis-ball mask.
I K.il)l)it hoard-.
ma>k.
M .it IT »x>ttles.
'IVnni- hall mask.
Bell.
izccl dog.
APPENDIX
Groups or
Demonstration.
Group IV
Exercise. Animals.
VII
Solutions.
Liu-tic Acid, 0.6% (20).
Caffein, i(o (20).
Camphor, i% in 40% Alcohol (10).
Strychnin, 0.1% (10).
Group V VIII Ammonia in blowing bottle.
Ammon. Chlorid, i% (150).
Strychnin, 0.1% (10).
Total Animals Needed. — 5 Rabbits (s); 2 morphinized dogs (f).
Groups or
Demonstration.
Group IV
Groups I, II,
III, and V
Group IV, A
Group IV, B
Groups I, II,
andV
Groups II, V
Groups II, III,
and V
Group V
Group III
CHAPTER XLIL— ANESTHESIA
Exercise.
Animals.
2 Rabbits.
Dogs.
II
III, IV
V
VI
Solutions.
Epinephrin, i : 1000 (i).
WannN.S.(coo).t
Chloroform in blowing bottle.
Cocain, 2% (5).
Nitrous oxid.
Chloroform (10).
Morphin, 4% (5).
Ethyl chlorid (2).
Ether in blow-bottle.
Morphin, 4% (2)-
Scopolamin, i : 1000 (2).
Chloroform (25).
Ether, sat'd in N. S. (25).
Morphin, 4% (5).
Scopolamin, i% (2).
Curare, |% (15).
Phenol, i% (100).
Epinephrin, i : 10,000 (200).
Special apparatus.
Total Animals Needed. — 2 Rabbits (s); 4 dogs (f).
Rectal catheter.
Oncometer.
Compressed air or
oxygen.
Apparatus for in-
sufflation, with
ether and chloro-
form.
Catheter.
Ca rdioplethysmo-
gram.
Saw.
Cautery.
Groups or
Demonstration.
All Groups:
Group I
Groups II, III
Group IV
Group V
Groups:
I, II
I, IV, V
I, III, IV, V
II, III, IV, V
II, IV
II
II
II
CHAPTER XLIIL— VASOMOTOR DRUGS
Exercise. Animals.
Morphin-
ized dog.
(Smallest
dog for
Group II)
Solutions.
Epinephrin, i : 10,000 (10).
Special apparatus.
Amyl Nitrite (2).
Nitroglycerin, i : 1000 (10).
Strophanthus, i : 100 (i).
Strychnin, i : 1000 (i).
Pituitary Sol'n (i).
Ergot, 25% (10).
Tyramin, i : 1000 (20).
Histamin, i : 10,000 (2).
Stephen Hale man-
ometer.
Oncometers.
Cardioplethysmo-
gram.
Vasomoton perfu-
sion.
APPENDIX G
INDIVIDUAL PHARMACODYNAMIC EXERCISES
319
Groups or
Demonstration.
Groups:
II
II
II
II
II
III
III
III
III
III
IV
IV
IV
V
V
V
V
Exercise. Animals.
Total Animals Needed. — 5 Morphinized dogs (f).
Solutions.
Cholin, i : 1000 (20).
Cotarnin, i : 100 (10).
Hydraslis. 2% (10).
llydra>tinin. i : 100 (5).
Nicotin, o.i Vc (s).
Sod. Nitrite, 10% (5).
Alcohol. 2595 (50).
Veronal. Sodium. 10' 'L (25).
Peptone, 10' < (50).
Ammonia in blow-bottle.
Phenol, i', (50).
Chloral, 10' < (50).
Arsenate Sodium, 5% (50).
Chloroform (10).
Caffein, i% (10).
Cevadin, i : 1000 (i).
Atropin. i : 1000 (i).
CHAPTER XLIV .-CHANGES IN
Groups or
Demonstration.
All Groups:
Exercise.
Group I
Group II
Groups I
III. IV
Group* I. V
Group II
Group II
Group II
III
Group IV
Group IV
p V
Group V
II,
Animals.
Morphin-
ized dog.
Solutions.
Cevadin, i : 1000 (i).
HEART-RATE, ETC.
Special apparatus.
Cardiomyogram.
Cardioplet h y smogram .
Total Animals Needed. — 5 Morphinized dogs (f).
Strophanthus, i : 100 (5).
Spartein, i : 100 (10).
Pilocarpin, i : 100 (2).
Di^itali*. 5 : 100 (too).
Ouabain, i : 1000 (i).
Atropin, i : 1000 (8).
Barium chlorid, i : 100 (30).
XitroL'lycerin, i : 100 (i).
Kpinephrin. i : 1000 (i).
CHAPTER XLV.— MYOCARDIAL DEPRESSANTS AND TONICS
Exercise. Animals.
Mprphin-
ized dog.
Groups or
Demonstration.
All Gr
Group III
Group I
p I
Group I
Group I. V
Group I. Ill
II. IV
II. IV
Group II IV
( iroup 1 1
Group 111
(iroup III
(Iroup III
(iroup IV. V
p V
p V
p V
Total Animals Needed—
Solutions.
v onite, 10' ( (15).
Phenol. i% (75).
(i).
CUorofo
\itroirl\-i erin, i% (2).
piiriii. i : looo (i).
;% (10).
;n Chlorid, i% (25).
CatTein, i% (25).
Spart.-in.
ili-. 5% (ioo).
•I»h;inthus 1% (O.
Str\-« hum. i ioo
ium . hlorid. i'"o ^50).
( 'a ni ph.. r. T , in 40% Alco-
Morj)hinized dogs (0-
Special apparatus.
Cardiomyoj»ram.
Cudioplethymognm.
320 APPENDIX
CHAPTER XLVI.— DIURESIS; CARDIAC LESIONS
Demonstration. Exercise. Animals. Solutions. Special apparatus.
All Groups: . . Morphin- Warm Saline (500).
ized dog. Strophanthus, i : 1000 (10).
Group I Cardioplethysmo-
gram.
Group V Cardiomyogram.
Group I Sod. Sulphate, 2.5% (400)
(dried).
Groups I, V Epinephrin, i : 1000 (i).
Group I . . Spartein, i% (10).
Groups II, III, NaCl, i% (25).
V
Groups II, III, MgSO4, 3-6% dried, (25).
V
Group II NaCl, i% (400).
Group II Theobromin Sod. Salic.,
10% (5).
Group III NaCl, 10% (40).
Group III . . Theophyllin, Sod. Acet.,
10% (10).
Group III . . Alcohol, 95% (15).
Group IV . . Glucose, 6% (400).
Group IV . . Amyl Nitrite (5).
Group IV . . Caffein, i% (15).
Group V . . Locke's Solution (no Glu-
cose) (400).
Group V . . Pituitary Solution (i).
Lycopodium Suspension
(10).
Total Animals Needed, — 5 Morphinized dogs (f).
APPENDIX H.— DOSES FOR ANIMALS
The drugs are arranged alphabetically; in the case of salts, by the more important
ion. In the case of crude drugs the dose refers to fluid preparations. The "just fatal"
doses have generally been worked out with considerable accuracy, but may vary some-
what with different samples of the poison and with each lot of animals. The doses
marked with an asterisk (*) have been confirmed by the author; the others were compiled
from pharmacologic literature.
Doses of drugs not contained in this list may be ascertained by consulting the original
papers cited in the Manual of Pharmacology.
M. F. D. = minimum fatal dose (average).
It is convenient to remember that a dose of i mg. per kg. corresponds to about 0.05 gm.
or i grain for an adult man.
Abrin.
M. F. D.: Rabbit, vein, per kg., o.oi mg.
Absinth.
Epileptic Convulsions: Dog, per kg., 0.03 to 0.05 c.c. of Essence (Ossipow, 1914, Ref.
Zbl. Bioch. Bioph., 17, 303).
Acetanilid.
Urine: Man, 0.2 gm. (Chap. 15, IX).*
Toxic Dose: Dog, stomach, per kg., 0.7 gm.: cyanosis and methemoglobinemia, fatal
in nine hours. Rabbit, stomach, per kg., 0.2 gm.: slowed heart and respiration;
paralysis of legs; recovery in three hours.*
Acetate, Sodium.
Urine: Man, iogm.: Alkaline (Chap. 15, VI).*
M. F. D. (usual): Dog, vein, per kg., 3 gm.
Not dangerous: Dog, vein, per kg., 35 c.c. of 1.94 per cent., crystals.* *
Acetphenetidin.
Urine: Man, 0.3 gm. (Chap. 15, IX).*
Acid, Acetic.
Fatal: Dog, stomach, per kg., 0.3 gm.
Reflex: Frog, 5 percent. (Chap. 32, III).*
i For fatal dose of a series of Sodium Salts, see Sabbatani.
APPENDIX II DOSES FOR ANIMALS 321
Acid, Hydrochloric.
Acido>i>: Rabbit' stomach, per kg., i gm. (looc.c. of i per cent.): slowed heart and
respiration, ascending paralysis, convulsions, death in twelve to forty-live min-
utes (Chap. 39, C).* Guinea-pig, rectum. 10 to 50 c.c. of i per cent.: slowed heart
and respiration, convulsions, fall of temj>erature, death by respiratory failure in
twelve to forty-nve minutes, early rigor.*
Acid, Lactic.
Me.lullarv Stimulation: Dog, vein, per kg., 2 c.c. of 0.6 per cent. (Chap. 41, VII, i).*
Acid Phosphate, Sodium.
>;<;/>. vein, 10 per cent. (Spiro).
Aconite.
Fatal Dose: Dog, hypodermic, per kg., 40 mg.: nausea, inco-ordinated movements,
irregular heart and slowed and irregular respiration, convulsions in twenty-five
minutes, death in thirty-four minutes.*
M. I . I).: (/'///H«J-/>/\'. hypodermic, i>ergm., 0.04 mg. (Chap. 36, III, 7).
;.u Arrest: Mammals, vein, per kg., 100 mg., i c.c. of 10 per cent.) (Chap. 45,
I, 5>-*
Therapeutic Dose: Mamtnais, vein, per kg., 5 mg. (^ c.c. of 10 per cent.) (Chap. 45,
Heart tracing: Frog, lymph-sac, 0.5 c.c. of 4 per cent. (Chap. 36, IV, 3).*
Aconitin (Cry-1
M. 1 I).: Dog, hypodermic, per kg., o.i mg. Rabbit, hypodermic, per kg., 0.5 mg.
(tuinra-pis, hypodermic, j>er kg.. 0.06 mg. (Merck's, 0.05 mg., Engelhardt).
Pigeon, hypodermic, per kg., 0.22 mg. Frog, hypodermic per gm., 0.016 mg.
Acrolein.
M. F. D.: Mammals, stomach, per kg., 0.15-0.2 gm.
Adalin.
Hypnotic : Dog. stomach, per kg., 0.25 gm. (Gensler, 1915).
Adrenalin. See F.pinephrin.
Albumose. See Peptone, \\ tile's.
Alcohol, Amyl.
Fall Blood-pressure: Dog, vein, per kg., 5 c.c. of 2 per cent. (Salant, 1909).
M. F. D.: Rabbit, stomach, per kg., 1.7 to 2.0 gm.
:• otic: Rabbit, stomach, per kg., 0.8-1.25 gin-
Alcohol, Butyl.
M I •'. D.: Rabbit, stomach, per kg., 2.1-2.5 g™-
Narrotir: Rabbit, stomach, per kg., 1.0-1.5 gin.
Alcohol, Ethyl.
Ergograph: Man, 20 to 40 c.c. of 20 per cent. (Chap. 33, V).
Ordinary Dose: Mammals, vein, per (4 c.c. of 25 per tent.; this, concentra-
tion does not precipit ate !>!<>< >d > (Chap. 4^. 111,4).*
M. F. D.: Rabbit, stomach, per kg., 6.25-7.25 gm. Cat, peritoneum, per kg., 8 c.c.
Paralytic Dose: Cat or Cninra-pJK. stomach or peritoneum, per kg., 4 c.c. (16 c.c. of
25 per cent.) (Chap ;o. XVI).* (Details, Pilcher, 1912, Jour. Pharmacol., 3,
267.) Rabbit, stomach, per kg., 5 c.c. (10 c.c. of 50 per cent.).* (Ami;
Antidote) stomach, per kg., 2.5 to 4 gm.: narcotic; recovery in one t
hours; 4.5 to6gm.: narcotic; recovery in >ix to ten hours.
Respiratory Stimulation: Rabbit, hypodermic, per kg.. 0.5 c.c. (i c.c. of 50 percent.)
-•).*
I )ose: Frogs, lymph sac, 2 c.c. of 25 p< ..; II, 3).*
Reflexes: Fro^ ac, 0.5 c.c. < f 10 per . ent. (Chap. 32, III, 2).*
Alcohol, Methyl.
M I h JbftMf, ttonach, per kg., 7-9 gm.
Narcotic: Rabbit, «.toma« h. per k-j.. J.3 5.5 gm. Dog, stomach, per kg., 4 C.C.
(sleep lasting several days).
Aleuron.
Exudatc: Mammals, pleural or In podt-rmii . 10 i •.« • -pension in 3 per
starch paste (Chap \\ I
Aloin.
•'•//. hy|MKlermic, per kg., 2 c.c. of 5 per \ \ I
Dog, hypodern * Nider, iui
Alum.
' Absorption: 0.25-0.5 per
Aluminum Salt* (Calculated as Metal).
Ml l> /'• tto, Cat, 150 mg.; ditto, Rabbit,
mg.
at
322 APPENDIX
r Ammonium Carbonate.
Emetic: Dog, stomach, 20 c.c. of 5 per cent. (Chap. 38, XVI).*
Convulsions: Rabbit, hypodermic, per kg., 0.4 gm. Frog, lymph-sac, '25 c.c. of i per
cent. (Chap. 32, 1, 9).*
Ammonium Chlorid.
Medullary Stimulation: Mammals, vein, per kg., 0.15 gm. (15 c.c. of i per cent.)
(Chap. 41, VIII, 2).*
Anesthesin.
M. F. D.: Dog, vein, per kg., 0.4 gm.; peritoneum, per kg., 0.75 gm.; ditto, Cat or
Guinea-pig, 0.9 gm. Rabbit, stomach, per kg., 1.15 gm.
Anilin.
Toxic: Frog, 2 drops in mouth.*
Antimonium Potassium Tartrate.
Emetic: Dog or Cat, stomach, 0.003 to o.i gm. (Chap. 38, XVI).* (Details, Pitten-
ger, 1913.)
Fatal: Rabbit, vein, per kg., 0.15 (6 c.c. of 2.5 per cent.), fatal in twenty-four hours.
Antipyrin.
Urine: Man, 0.3 gm. (Chap. 15, X).*
Antipyretic: Rabbit or Cat, hypodermic, per kg., o.i gm. (i c.c. of 10 per cent.)
(Chap. 39, VI).* Rabbit or Cat, stomach, 0.5 gm. (Gottlieb, 1890).
Ordinary Dose: Mammals, vein, per kg., o.i gm. (i c.c. of 10 per cent.) (Chap. 45,
I, 2).*
M. F. D.: White Mice, hypodermic, per gm., i mg. (Hale, 1910).
Apocodein Hydrochlorid.
Xerve Paralysis: Mammals, vein, per kg., 40 to 50 mg. (as i per cent.). Local, i per
cent. Perfusion, inject 2 c.c. of i per cent.
Apocynum.
Emetic: Dog, stomach, per kg., 0.2 gm.*
Cardiovascular: Dog, hypodermic, per kg., 0.35 gm.*
M. F. D.: Frog, lymph-sac, per gm., 0.05 mg.
x Apomorphin Hydrochlorid.
Emetic: Dog, hypodermic, per kg., i mg. (o.i c.c. of i per cent.); effective in two
and one-half to ten minutes (Chap. 38, XIV)*; just effective, 0.2 mg. per kg.
(Eggleston and Hatcher, 1912; also with other methods of administration).
Cat, much higher.
Hypnotic: Dog, hypodermic, per kg., 0.04 mg. (often unsuccessful).*
Excitant: Rabbit, hypodermic, iomg.*
M. F. D.: White Mice, per gm., 0.4 mg. (Hale, 1910).
Arsenate, Sodium.
Cardiovascular: Mammals, vein, per kg., 50 mg. (i c.c. of 5 per cent.) (Chap. 43, IV,
10).*
Nephritis: Rabbit, hypodermic, per kg., 10 to 35 mg. (1-3.5 c.c. of i per cent.) (Chap.
39, XXI).* Dog, hypodermic, per kg., 1-20 mg. (MacNider, 1912).
Enteritis: Rabbit, hypodermic, per kg., 50 mg. (i c.c. of 5 per cent.) (Chap. 39, XVIII,
Fatal: Rabbit, hypodermic, 5 c.c. of 5 per cent.*
Arsenic Acid.
M. F. D.: Rabbit, hypodermic, per kg., 12.4 mg.; fatal in two and one-half days
(Kionka, 1911).
Arsenic Trioxid.
M. F. D.: Rabbit, hypodermic, per kg., 8.33 mg.; fatal in four days (Kionka, 1911).
Arsenite Potassium, Liquor (Fowler's Solution).
Effects: Rabbit, stomach, 0.6 c.c. per kg., may be fatal inside of twelve hours; 1.5 c.c.
may be survived. Dog, i c.c. per kg., vomiting, may be fatal; 3.5 c.c. may be
survived; if vomiting is prevented by morphin, 0.5 to i c.c. may be fatal inside
of twelve hours.
Aspidospermin.
Respiratory Stimulation: Dog, hypodermic or vein, per kg., 2.5 to 8 mg.
Atoxyl.
Fatal, Dog, hypodermic, per kg., 20 mg. (Details, Dietrich, 1910, Merck's Rep.,
24, 117.)
< Atropin Sulphate.1
M. F. D.: Rabbit, per kg.: stomach, 1.4-1.5 gm.; hypodermic, 0.5-0.75 gm.; vein,
0.07-0.075 gm. Dog, per kg., hypodermic, 0.14-0.4 gm.; vein, 0.06-0.07 gm.
Cat, per kg.: hypodermic, 0.03 gm. Guinea-pig, per kg.: hypodermic, 0.6 gm.;
vein, 0.085 gm- R°t, Per kg.: hypodermic, 2.5 gm.
i Doses for different animals: Cloetta, 1905; Heffter, 1911. Wilberg, 1914.
APPENDIX H DOSES FOR ANIMALS 323
» Atropin Sulphate (Continued).
Mydriatu : Cat, per kg.: stomach, 0.5 mg.; hypodermic, 0.04 mg.; vein, 0.02 mg.;
rectal, 0.7 mg. iHauhcr and Kggleston, 1914).
Respiratory stimulant: Rabbit, hypodermic, per kg., i mg. (i c.c. of i : 1000) (Chap.
41. V;.*
Antagonist to Pilocarpin (intestines and bronchi, etc.): Rabbit, vein, per kg., 1-2
mg. (i : i : ioooj (Chap. 34, 1, 10; Chap. ;;. VII .*
Antagonist to Cholin: Mammals, vein, per kg., i mg. (Chap. 43, II, 8).*
is: Dog, vein, per kg., 0.05 mg. & c.c. of i : 1000) (Chap. 44, IV, 4).*
(Details, Sollmann and Pilcher, 1914.)
Dog, hypodermic, per kg., 0.2 mg., not toxic; effect on pupil and
heart (*); 10 mg., vomiting (.*); 40 to 80 mg., severely toxic, but usually not
fatal.* M. I . I). lie- between 20 and 400 mg. Cat. hyi>odermic. mg. j •
0.02, no effect on pupil; 0.04, good dilatation; 0.05 just paralyzes vagi. Frog,
lymph >u« . i mg., little effect; 2 mg. motor depression with recovery; 10 mg.
per 20 gm., fatal; 50 mg., ordinary dose; 100 mg., fatal (*).
Barium Carbonate.
Fatal: Dog, stomach, 4 gm.
Barium Chlorid.
Cardiovascular: Mammals, vein, per kg., 20 mg. (2 c.c. of i per cent.) (Chap. 44,
IV- 5)-*
Peristalsis: Rabbit, vein, per kg., 10 mg. (i c.c. of i per cent.) (Chap. 34, I, n).*
Benzoic Acid.
Fatal: Frog, lymph-sac, per gm., 2.5 mg. (three hours) (Impens).
Berberin.
:iovasc -ular: Mammals, vein, per kg., 1-5 mg. (Chap. 43, 1, 15)* (Williams, 1908).
; session: Mammals, vein, per kg., 2-20 mg. (Marfori, 1890).
Vagus Paral>>i>: Frog, lymph-sac, 10 mg. (Marfori, 1890).
Beta-tetra-hydronaphthylamin.
Pyretic: Rabbit, hvjxxlermic, per kg., 25-50 mg. (\ to i c.c. of 5 per cent.) (Chap.
39, V).* (Details. C'oleman, 1907; Ott and Scott, 1907; Jonescu, 1909.)
Bichromate, Potassium.
Fatal: Dog, stomach, 0.06 to 0.4 gm.
Bismuth Subcarbonate.
Antemet tomach, i gm. (Chap. 38, XVIII).*
Bromid, Sodium.
Antispasmodic: Cat or Rabbit, stomach, per kg., 2 gm. (10 c.c. of 20 per cent.) (Chap.
40, I \
Bromural.
ts: Cats, stomach, per kg.: Mean fatal, 0.45-0.5 gm.; Deep coma, 0.4 gm.;
Light coma, 0.25-0.3 gm.; Sound natural sleep, 0.1-0.15 K01- (Sollmann and
Hati her, 1908).
Hypnotic: Dogs, stomach, per kg., 0.25 gm. (Gensler, 1915).
Brucin Hydrochlorid.
Hypodermic, per kg.: Dog Rabbit Pigeon Mouse
Inert ; mg. 6.25 mg. 6.O mg.
Convulsive 4.5 mg. 7.5 mg. 26.5 mg. 40.3 mg.
7.5 mg. 8.6 mg.
M II) 18.5 mg. 42.2 mg. 108.2 mg.
Dot. -kg.: 4.25-1(1 mg., increased rcll. tank.
Cadmium Salts.
.irgc). hyixHlermic, 0.3 gm. Rabbit. >tomaih. 0.02-0.04 gm. Frog,
lymph -.t. . per irm.. i mg.
. Cafle.n I'M, \!k,,],
p \ '
('in illation and Kc-; ! \nlid. -tc M,:n:n:
10-20 mg « \\ I. \\ 11
Ch.tp. 41. I. c; Chap. ;i. \ II
19.)
XT kg., 40-100 mg. (4-10 >
.\ I '
I and kigi.r: Fru,;. lymph-sac, 10 mg. (i c.c. of i per cent.) (Chap. 32,
I. i
5 mg. (§ c.c. of i p< III, 5).*
ta: Frog, |-i c.c. of i per I i ?).*
324 APPENDIX
Caffein (Continued).
Fatal Dose, per kc:.: Vein Hypodermic Stomach
Dog o.i i ,urni. 0.14 gm.
(sometimes) (sometimes)
Cat. . . '. 0.15 gm.
Rabbit . . 0.16 gm. 0.28 gm. 0.36 gm.
Guinea-pig.. 0.28^111.
(Salant and Rieger, 1910.)
Calabarin.
Fatal Convulsions: Rabbit, hypodermic, per kg., 20 mg.
^ Calcium Chlorid.
M. F. D. Dog, vein, per kg., 0.444 gm. (4 c.c. of m/8) (Joseph and Meltzer, 1909).
Antagonism to Mg.: Rabbit, vein, 0.180-0.240 gm. (6-8 c.c. of 3 per cent.) (Chap. 39,
XV).*
Coagulation Time: Rabbit, hypodermic, o.i gm. (i c.c. of 10 per cent.): effect in one
to three hours (Coleman, 1907).
Calcium Lactate.
Airuinst Inflammation and Effusions: Dog or Cat, hypodermic, per kg., 20 mg. (2 c.c.
of i per cent.) (Chapter 37, XII and XIII).*
Calomel.
Purgative: Dog, stomach, per kg., 0.16 gm. (Valeri, 1909).
Systemic Effects: Dog, stomach, per kg., 0.21 gm. (Valeri, 1909).
Diuresis and Enteritis: Rabbit, hypodermic, per kg., 5-10 mg. (1-2 c.c. of 0.5 per
cent, in sod. thiosulphate).
Camphor.
Stimulant Dose: Mammals, vein, per kg., 5 to 30 mg. (£ to 3 c.c. of i per cent, in
40 per cent, alcohol) (Chap. 41, VII, 3; 45, V, 4).* (Details, Gottlieb, 1905.)
Rabbit, hypodermic, per kg., o.i gm. (| c.c. of 20 per cent.) (Chap. 41, I, 4).
Convulsant, Ordinary Dose: Cat or Rabbit, stomach, per kg., 2 gm. (10 c.c. of 20 per
cent, in oil) (Chap. 40, III).* Frog, o.i gm. (i c.c. of 10 per cent.) (Chap. 32,
I, 9) (also Curare Action, Chap. 32, IV, 10).
Minimum Convulsant Dose, per kg. : Stomach Hypodermic Intramuscular.
Dissolved in oil : Dog 0.5 gm. 0.75 gm. 0.5 gm.
Dissolved in alcohol: Dog. ... 0.5 gm. 1.5 gm. 0.75 gm.
Dissolved in oil: Cat 0.25 gm. 0.5 gm. 0.5 gm.
Dissolved in alcohol: Cat 0.5 gm. 0.5 gm. 0.7 gm.
(Hatcher and Eggleston, 1914, Jour. Amer. Med. Assoc., 63, 469.)
M. F. D.: Frog, lymph-sac, per gm., 3.2 mg. (as 10 per cent, in oil) (Grove, 1910).
Guinea-pig, mouth, per 100 gm., 0.15-0.18 gm. (Cairis, 1914, Jour. Pharm.
Chem., 10, 224; also other methods of administration.)
Cane-sugar.
Comb: Rooster, pectoral muscles, per kg., 10 gm.: blueing in one-quarter to one-half
hour.
Cannabis.
Narcosis: Dog, stomach, per kg., 0.05 gm. of Extract (Chap. 39, XIV).*
Test Dose: Dog, stomach, per kg., 0.004 gm. of Extract (Chap. 39, XIV), Dog,
stomach, per kg., 0.03 gm. of Fldext.
Ataxia: Cat, hypodermic, £ c.c. of Tinct. with \ c.c. water (Dixon).
Cantharidin.
Vesicant: Man, 0.15 mg. local.
Nephritis: Mammals, hypodermic or vein, per kg., i to 10 mg. (in acetic ether),
severe (Chap. 39, XXI).* Glomerular only, o.i to i mg. per kg.
Caramel.
Antidote: Cat, stomach, logm. (Chap. 40, VII).*
Cascara.
Urine: Man, i c.c. of Fldext. (Chapter 15, XII).*
Cephaelin.
M. F. D.: Mammals, hypodermic, per kg., 30 mg.
Emetic: Dog, stomach, per kg., i mg.
Cerium Oxalate.
Innocuous: Rabbit, stomach, per kg., 0.7 gm. (Bachem, 1907).
Cevadin. See also Veratrin.
Vagus Stimulation: Mammals, vein, per kg., 0.05 mg. (fa c.c. of i : 1000) (Chap. 44,
1,3)-*
:ent
APPENDIX II DOSES FOR ANIMALS 325
ogy
Cevadin (Continued).
Convulsive: Rabbits, hypodermic, per kg., 3 mg.*
M. F. D.: Rabbit and Guinea-pig, hypodermic, per kg., 3-6 mg.*
Fatal: Rabbit, stomach, per kg.. 10 mg.*
Charcoal.
Antidote: (.'.;/. stomach, 10 gm. (Chap. 40, VII).*
Chloral.
Ordinary Dose (Anesthetic, nanotii , temperature, antidote, etc.): Cat, stomach, per
kg., 0.25 gm. (2.5 c.c. of 10 per cent.) (Chap. 39, I, XVII; 40, X).* Dog,
stomach, per kg., 0.25 to 0.3 gm.; vein, per kg., o.i to 0.15 gm. (Chap. 41. TV
Rabbit. >tomach. per kg., 0.6 gm.*; rectum, per kg., 0.3 gm. (Chap. 41, IV, 3).*
Successive Effects: Cat, stomach, per kg., 0.09 to 0.15 gm.: natural sleep; 0.18 to
0.25 gm.: light coma: 0.30 gm. up: deep coma; 0.42 to 0.45 gm.: mean fatal dose.
(Sollmann and Hatcher, 1908.)*
Circulatory Depression: Mammals, vein, per kg., 0.5 gm. (5 c.c. of id per cent.)
(Chap. 43, IV, 8).*
Respiratory Depression: Rabbit, stomach, per kg., 0.5 gm. (20 c.c. of 2.5 per cent.)
(Chapter 41, IV, 3).*
"ti< : Frog, lymph-sac, 0.02 gm. (i c.c. of 2 per cent.) (Chap. 32, II, 4).*
Cardiac Depression: Frog, lymph-sac, 0.04 gm. (0.4 c.c. of 10 per cent.) (Chap. 36,
V, i).*
Fatal: Frog, lymph-sac, o.i gm. (i c.c. of 10 per cent.).
x Chloralosc.
Anesthetic: Dog, stomach, per kg., o.i gm. (Pawlow).
Chlorate, Potassium.
Fatal: Rabbit, stomarh, per k^., 4 pn.: methemoglobin cyanosis, respiratory paral-
\ -is. convulsions, death in lour hours.
Chloretone.
Anesthetic (after morphin): Dog, stomach, per kg., 0.2 to 0.25 gm. (in alcohol) (Chap.
41, TN).* Cat, stomach, per kg., 0.3 gm. (in alcohol) (Chap. 41, TN).* Rabbit,
stomach, per kg., 0.15 to 0.2 gm. (in alcohol) (Chap. 41, TN *
Circulatory Depression: Dog, vein, per kg., 0.5 c.c. of saturated watery solution.*
., Chlorid, Sodium.
me Infusion: Mammals, vein, per kg., 25 to 100 c.c. of 0.9 per cent.*
Diuretic: Mammals, vein, per kg., 2.5 c.c. of 10 per cent. (Chap. 46, III, 2).*
Fatal : Mammals, vein, per kg., 10 to 30 c.c. of 10 to 33 per cent.; death in four to five
minutes.* Dog, vein, per ktf., 64 c.c. of m/8. (Joseph and Melt/er. 1909.)
^ Chloroform.
Ane-thetif: Mammals, vein, per kg., i c.c. of 0.5 per cent. (Chap. 4- ' \'-
Narcotic: Frogs, lymph-sac, 0.2 gm. (i c.c. of 20 per cent, in oil) (Chap. 32, II, 6).*
Fatal: Frogs, lymph-si*-, 0.45 gm.
Kidney Lesions (Fiske and Karsner, 1914).
^holin.
Ordinary Dose: Mammals, vein, per kg., i to 2 mg. (i to 2 c.c. of i : 1000) (Chap. 43,
II. 8).* (Details. Abderhalden and Mueller, 1911: Busquet and Puthon. 1012).
Chromate, Potassium. See Hi< Inornate.
Fatal: Rabbit, hyi>odermir o.j-o.4 gm.; stomach, 2 gm.
Nephritis: Dog, hypodermi.. per k^., 2.5-50 mg ; vein. ; mg. (MacNidcr.
1012). Rabbit, hypodermi. , per kg., 30 mg. U'hap. .19, XXI).* Guinea-pig,
hy|Nidermi«. per ktf.. 50 mg. (nearly fatal. npluil>. KM
•"sits: Pigeons, 10 mg.; Hens, 10 to 20 mg. (hyjxxlermii. repeated sc\er.«l
da\
Citrate. Sodium (CrystalsJ.
Trim-: Man. -t..m;i. li. 10 gm.. alkaline reai lion (Chap
M.I- l> 7>«>c. veii <; Kin. Fro£, lynip'1 nig.
Cobalt, Nitrate.
Mi I' , bypodomlc, per kg ., (o to 75 oof . P/V
t.. 10 rm- |H,iU-rmi. . |«-r ki: . i.S to 4 mg.
• rid.
<-s for mammal-, (irode, 1912. '
-;.i/j, hy|x)d- rkg., .'S mg. (o.s
MI l» K :'" .0.1 to O.i -i. 0.01 to 0.022 ^
in, 0.015 to o . 0.05 to 0.06 gin.
Frog, lymph-sac, 3 i
al Notes.
326 APPENDIX
Cocain Hydrochlorid (Continued).
Intravenous Anesthesia: Rabbit, ear vein, per kg., 10 mg. (i c.c. of i per cent.)
(Kilter, 1909; Chap. 32, V, n).
Effects: Dog, hypodermic, 2.5 mg. X kg., raises temperature by 0.2° to 0.5° C. for
two hours; 10 mg. X kg. by i° to 2° for three to four hours; 20 mg. X kg., 2° to 4°
for six to seven hours; 15 to 20 mg. X kg. emesis, mydriasis, convulsions, and
paralysis, with recovery; 25 mg. X kg., sometimes fatal; 80 mg. X kg., some-
times recovery. Rabbit, hypodermic, 20 mg. X kg., ordinary dose for hyper-
pyrexia (0.25 to 0.8° in one to three hours) (*); 30 mg. X kg., slight trembling;
50 mg. X kg., considerable rise of temperature; 60 mg. X kg., convulsions,
paralysis, recovery; 100 mg. X kg., sometimes fatal; 130 mg. X kg., sometimes
recovery; 540 mg. X kg., surely fatal.
Cocculus.
Fatal:. Dog, hypodermic, per kg., 0.4 gm.
Convulsant: Frog, lymph-sac, per gm., 2 mg.
v Codein.
Respiration: Mammals, hypodermic, per kg., 5 to 10 mg.
M. F. D.: Cats, hypodermic, per kg., 60-90 mg. (Mueller, 1908). Rabbit, hypodermic,
per kg., 60 mg.
Toxic: Frog, lymph-sac, 10 mg. (i c.c. of i per cent.) (Chap. 32, II, 6).*
Colchicin.
Leukocytosis: Rabbit, hypodermic, per kg., 5 mg. (i c.c. of 0.5 per cent.); maximum
in twelve hours (Coleman, 1907).
M. F. D.: White Mice, per kg., 1-25 mg. (Fuehner, 1910).
Colchicum (Seed).
(Colchicum root requires about twice these doses).
Fatal Gastro-enteritis: Dog or Cat, stomach, per kg., 0.5 c.c. of Fldext. (Chap. 39,
XVIII, i).*
Colocynth.
Purgative: Cat, stomach, 10 c.c. of 10 per cent, decoction: liquid stools in one to four
hours.
Coniin.
Fatal: Cat, hypodermic, 0.4 gm., fatal in one hour; 0.05 gm., fatal in nine hours.
Rabbit, hypodermic, per kg., 90 mg. Pigeon, hypodermic, per kg., 40 mg.
Mouse, hypodermic, per kg., 75 mg.
Paralytic: about three-quarters of the fatal dose.
Curare Action: Frog, lymph-sdc, 10 mg. (i c.c. of i per cent.) (Chap. 32, IV, 10).
Conium.
Ineffective: Dog, hypodermic, per kg., 0.5 gm.*
M. F. D.: Guinea-pig, hypodermic, per kg., 0.5 gm.* White Rat, hypodermic, per
kg., 40 gm.* Frog, lymph-sac, per gm., 0.06 gm.*
Convallaria.
M. F. D.: Guinea-pig, hypodermic, per kg., 0.08 gm. Rat, hypodermic, per kg.,
32 gm. Frog, hypodermic, per gm., 0.18 mg.
Copaiba.
Urine: Man, i gm.*
Copper Sulphate.
Emetic: Cat, stomach, 25 c.c. of i per cent. (Chap. 38, XVI).* Dog, stomach,
50 c.c. of i per cent.*
Coriamyrtin. *
Convulsive: Dog, hypodermic, per kg., 0.15 nig. Rabbit, hypodermic, per kg., 0.75 mg.
Fatal: Cat, hypodermic, per kg., 0.25 mg. Guinea-pig, hypodermic, per kg., 2.5 mg.,
Frog, hypodermic, per kg., o.i mg.
Cotarnin, Hydrochlorid or Phthalate.
Circulatory: Mammals, vein, per kg., 5 mg. (£ c.c. of i per cent.) (Chap. 43, II, 9).*
y Curare.
Paralytic: Mammals, vein, per kg., 3 mg. (f c.c. of \ per cent.), repeated every ten
minutes as needed (Chap. 32, IV, 6; 42, VI, 3).* Frog, lymph-sac, 2.5-5 mK-
(| to i c.c. of 5 per cent.) (Chap. 32, IV, i)*; immersion of muscle: jV per cent.
(Chap. 32, IV, 4)-*
Curarin.
Paralytic: Mammals, vein, per kg., 0.5 to 3 mg. Frog, lymph-sac, per gm., 0.00025
to o.ooi mg.
Cyanid, Potassium.
Toxic: Mammals, vein, per kg., 5 mg. (£ c.c. of i per cent.) (Chap. 43, I, 15).*
M. F. D.: Rabbit, hypodermic, per kg., 1.9 mg. (no effect below i mg.). Mouse,
hypodermic, per kg., 4.4. mg. Pigeon, hypodermic, per kg., 1.5 to 2.4 mg.
APPENDIX H DOSES FOR ANIMALS 327
Cystisin.
Fatal: Cat, hypodermic, 30 to 40 mg.
Delphinin (Heyl).
Vagus Paralysis: Rabbit, hypodermic, 75 mg. (1.5 c.c. of 5 per cent.).
Fatal: Dog or Cat, 0.03 to o.i gm.
.- Digitalis.
C irculatory, Ordinary: Mammals, vein, per kg., 50 mg. (i c.c. of 5 per cent.) (Chap.
45, III, 4).*
Circulatory, Toxic: Mammals, vein, per kg., 100 mg. (2 c.c. of 5 per cent.) »
111,4).* Frog, lymph-sac, 25 mg. (0.5 c.c. 015 per cent.) (Chap. 36, 1\ '
M. F. D.: Frog, lymph-sac, per gm., 0.6 mg. (Chap. 36, III).* (Hatcher, 1912):
Cat, vein, per kg., o.i gm. Dog, vein, per kg., 0.125 gm. Rabbit, vein, per kg.,
0.2 to 0.25 gm.
Digitaloid Drugs.
M I . D.: Cats, Frogs, and Guinea-pigs (Chap. 36, III). Cats, vein, (Hatcher, Anli.
Int. Med., Sept., 1912).
Digitoxin.
M. F. D. (Hatcher, 1912): Cat, vein, per kg., 0.3 mg. Dog, vein, per kg., 0.5 mg.
Rabbit, vein, per kg., 0.75 to i mg.
Dionin.
Respiration: Rabbit, hyixxlermk, per kg., 6 mg.
Fatal: Rabbit, hypodermic, per kg., 100 mg.
Local, F\e: 10 per cent.
Diphtheria Toxin.
. er, Arch. exp. Path., 60.)
Diurctin. See Tlicobromin.
Emetin.
Just Emetic: Dog, hypodermic, per kg., il mg.; vein, 5 mg. per kg.
Fatal: Mammals, hypodermic, per kg., o.i gm.; vein, per kg., 0.02 gm. Frog, lymph-
sac, 10-20 mg.
Paralysis: Frog, lymph-sac, 5 mg.
L«>«al: Dog's conjittu •//;•<;, i : 500; irritant.
Epinephrin (Adrenalin).
Blood-pressure: Ordinary Dose: Mammals, vein, per kg., 0.02 to 0.05 mg. (^ to ^
c.c. of i : 1000) (Chap. 43, I, 9).*
Minimal Rise: Atropinized dog, vein, per kg., o.oooi mg (Toujan, 1905). Rabbit,
vein, per kg., 0.0003 nig- (Cameron, 1905).
Maximal Rise: Cat, vein, per kg., 0.03 mg. (Elliott, 1905). Rabbit, vein, per kg.,
0.047 ing- (Pruszynski, 1905).
Progressive Rise: Atropinized dog, vein, per kg. (Hunt, 1901, and others):
0.000085 mg. =
5
mm. rise.
0.00025
mg.
•
7
mm.
H
0.0005
mg.
•
15
mm.
0.0007
mg.
20
mm.
0.0017
mg.
i
25
mm.
0.004
mg.
-
45
mm.
0.0055
mg.
65
mm.
0.03
mg.
150
mm.
Bromhial Relaxation: Rabbit, vein, o.i n t i : 10,000) (( ! VM *
:'»/•//. hy|M.<lc-rnii( , i to : r \ '
M>'<" "i" i : 100. mi:. of Jv
Sulphate «.r «-i Cam-in.
Pupil: /T«S ' i : 1000 (Melt/er and \iier. 1004).
M. I 1 1 o.i to o.. :.< . PIT kj:..
in. per kg.. 0.5 to o.S mg
0.2 too.f. r :«Tmii . per kur.. .'.5 to ionm. il'atun. njoj. I
pig, • .:.. o.i to o.: n rmi« . |»et '.
Ergot.
, 0.025 to 0.25 gm. < ,
R00>!' '
190
Uterus: ( th, 1^08).
M.I I' urn., 50 mp.
328 APPENDIX
Ergotoxin.
Circulation: Mammals, vein, per kg., 0.25 to 0.5 mg. (i to £ c.c. of i : 1000) (Chap.
43, I, 15)-
Ether.
Anesthetic: Mammals, vein, per kg., i to i c.c. of saturated solution (Derouaux,
1909) (Chap. 42, III, 14).*
Stimulant: M annuals, hypodermic, 5 c.c.*
Eucain, Beta-, Hydrochlorid.
M. F. D.: Rabbit, hypodermic, per kg., 0.4 to 0.5 gm. (Vinci). Guinea-pig, hypo-
dermic, per kg., 0.3 to 0.35 gm. (Vinci).
Ferrocyanid, Sodium.
Non-toxic: Dog, vein, per kg., 35 c.c. of 7.5 per cent, crystals.*
Filmaron.
Anthelmintic: Dogs, 0.2 to i gm., in capsules, followed by purgative (Gmeiner, 1907;
Merck's Rep., 21, 108).
Fluorid, Sodium.
M. F. D.: Dog, vein, per kg., 0.05 to o.i gm.; hypodermic, per kg., 0.15 gm. Rabbit,
vein, per kg., 0.14 gm.; hypodermic, per kg., 0.15 gm.; stomach, per kg., 0.5 gm.
Frog, lymph-sac, 40 mg.
To kill epithelium, 0.03 to 0.3 per cent.; preservative, 0.2 per cent.; muscle twitchings,
0.5 per cent.
Formaldehyd.
M. F. D.: Dog, vein, per kg., 0.07 gm.; hypodermic, per kg., 0.35 gm. (twenty-four
hours). Rabbit, vein, per kg., 0.09 gm.; hypodermic, per kg., 0.22 to 0.5 gm.
(several days). Guinea-pig, hypodermic, per kg., 0.8 gm. Frog, lymph-sac,
0.8 mg.
Formate, Sodium.
M. F. D.: Dog, stomach, per kg., 4 gm.; vein, per kg., 3 gm. Rabbit, somewhat
larger (Fleig, 1907).
Fuchsin, Acid.
Convulsions: Frog, lymph-sac, per gm., 0.03 c.c. of 5 per cent. (Chap. 32, 1, 7).*
Gelseminin Hydrochlorid.
Toxic: Frog, lymph-sac, 20 mg.
Gelsemium.
M. F. D.: Guinea-pig, hypodermic, per kg., 1.75 to 6 gm. White Rat, hypodermic,
per kg., 2.2 gm. Frog, lymph-sac, per gm., 6.5 to 15 mg.
Glucose.
Diuretic: Mammals, vein, per kg., 25 c.c. of 6 per cent. (Chap. 46, IV, i).*
Glycerin.
Muscular: Frog, lymph-sac, 0.5 to i c.c.
Gold (Calculated as Metal).
M. F. D.: Dog, hypodermic, per kg., 0.4 gm.; ditto, Cat, 0.45 gm.; ditto, Rabbit,
0.36 gm.; ditto, Frog, 0.30 gm.
Grehant Anesthetic.
Anesthetic: Dogs, per kg.: Morphin, hypodermic, ? of 4 per cent.; Grehant Mixture,
stomach, 6 to 10 c.c. (Chap. 41, TN).* (Mixture contains 5 per cent, of chloro-
form in 50 per cent, alcohol.)
Guanidin.
(Fuehner, 1907, Arch. exp. Path. Pharm., 58, i.)
Hedonal.
Rabbit, stomach, per kg.: sleep, o.i to 0.2 gm.; anesthetic, 0.25 gm.; toxic, 0.35 gm.
(Cataldi, Wien. med. Presse, 1906, No. 50.)
Helleborein.
M. F. D.: Frog, lymph-sac, per gm., 0.004 nig.; ditto, Toad, 0.185 to 0.244 mg.
Helleborus Niger.
M. F. D.: Guinea-pig, hypodermic, per kg., 0.2 gm.; ditto, White Rat, 20 gm.; ditto,
Frog, 0.3 gm.*
Heroin.
Respiration: Rabbit, hypodermic, per kg., 0.5 mg.*
Hexamethylenamin.
Excretion: Man, stomach, 0.5 gm. (Chap. 15, V).* Dog, stomach, per kg., 0.5 gm.
(Chap. 38, IX).*
M. F. D.: Guinea-pig, hypodermic, per kg., over 10 gm. (Frothingham, 1909).
> Hirudin.
Coagulation: Mammals, vein, per kg., 20 to 50 mg. Direct addition to blood,
i : 6000.
APPENDIX H DOSES FOR ANIMALS 329
X Histamin.
Bronchi and Circulation: Mammals, vein, per kj;., o.oi to o.i me. (A to i c.c. of
i : 10,000) (Chap. , II . ;).*
Fatal: Rabbit, vein, per kg., 0.5 mg. (Oehjne, 1913).
flordenin.
M I . D.: Dog or Rabbit, vein, per kg., 0.25 gm. (Martinet, 1910).
Hydrastin Hydrochlorid.
emulation: Mammals, vein, per kg., 5 mg. (5 c.c. of i : 1000) (Chap. 43, I, 15).*
\\ illiai Marfori, 1800.)
Convulsions: /•><»$;. lymph-s;u . i to _> c.c. of i : 1000 (Chap. 32, I, 9).*
Hydrastmin Hydrochlorid.
Circulation: Mammals, vein, per ktf.. i to 5 mg. (^ to $ c.c. of i per cent.) (Chap.
11. M ' Marfori, 1800; Williams, 1907.)
Pararyti l-r^. lymph-sac. 5 m^.; fatal, 15 mg. (Marfori, 1890).
Hydrastis.
Cin ulation: Mammals, vein, per k^.. jo mg. (i c.c. of 2 per cent.) (Chap. 43, II, 10).*
(Details, Williams. iqoy.)
InetTective: Dog, hypodermic, per kg., 0.2 gm.; ditto, stomach, 0.5 gm.
\uUi\e: h'rog. lymph-sac, i c.c. of 5 percent.
Hydrazin Sulphate.
M! Convulsions: Rabbit, hypodermic, per kg., 0.315 gm.
Kidney Le>i«m>: Cat, hyixxlermu , per kg., o.i gm., forty-eight hours before use
-ke and Karsner, 1914).
Hydrocyanic Acid (9 of Cyanid «>i" IN.ta-sium).
M 1 I ' Cat. per kg., 0.8-1 mg. (Lehmann).
Surely Fatal: Cat. stomach, per kg., 2 mg. (2 c.c. of i : 1000) (Chap. 40, VI.
r Rabbit. mouth, i c.c. of 2 per cent.; ditto, Dog, 5 c.c. of 2 per cent.
;i.2j.*
Hyoscin. See Scof>olamin.
„ Hyoscyamift.
MvJ^sis. Cat. hypodermic, per kg., 0.02 mg.
Mi^Funi \"a«u> I'araly>i>: C'<;/, hypodermic, per kp., 0.025 mg.
TOXM:: H "////<• Miff, hypodermic, per 12-15 gni., 10 mg.
I atal: \Vhitc Mice, hypodermic, per 12-15 K01-? 20 mg.
M<>tor depression: I''rog, lymph-sac, |>er 10 £m., j mg.
1: l:ro£. lympl' o gm., 10 m^r.
Hyoscyamus.
M.I I) Guinea -/>/£, hyjxxlermic, per kg., rogm.* Frog, l>rmph-sac,pergm., iomg.*
Hyposulphite. Sec ThiosHlph
lodid, Potassium.
:cti<»n: Man, mouth, o.,^ gm. (C'hap. 15, I).*
lodid. Sodium.
, \cin. per k^., 35 c.c. of 2.2 per cent.*
IMrural ctTu-ion: Do£, vein. p. • to per cent. (Chap. 37, XIII).*
Dei ire-ion: Rabbit. -t<>ma. h. 50 . .. . <>i i |>er cent.*
;/•/'//. hy|Midermi« . : « .» . «»f tincture.
X.ibbit. hy|H»(lermic, 0.075 Km-
lodoform.
Hypnoti, Rabbit, hypodermic, per k>:.. I mn in oil.
RMnt, stuma' h. i to .• urn.
Ipecac.
I .' h. per k«.. 0.2 to o..^ ^'m. (Chap. .;S, X\'I).
Iroi
Ml I' />.<.;. hyp...lenni. . 2 to 50 mg.; dit '. 25 mg. Frog, lymph-SEC,
per V
Isopral.
(Sollmann and Ilatiher, 1908) mral sleep,
; «m.: lik'ht «oma, o.n to o.iS gni.; deep ( onia, above 0.18 gm.; mean M.
1 1 » . to 0
Juniper Oil.
;
Laudanin.
Con- '. hyjuMlrr: . ;o mi:.
Ml I ' /)/•!;. hyjH,, ^o nig.
33° APPENDIX
Laudanosin.
M. F. D.: Rabbit, hypodermic, per kg., 68 mg.
Lead Acetate.
Intestinal Spasm: Mammals, vein, per kg., 5 to 8 mg. (Hirschfelder, 1915).
Leeches.
Coagulation: Mammals, vein, per kg., 3 heads in 6 c.c. of normal saline.
Lobelia.
Circulatory: Dog, hypodermic, per kg., 0.33 gm.*
M. F. D.: Guinea-pig, hypodermic, per kg., 10 gm.* Frog, lymph-sac, per gm.,
55 mg-*
Lobelin Sulphate.
Respiratory Stimulation: Rabbit, hypodermic, per kg., 2 mg.
Phrenic Paralysis: Rabbit, vein, per kg., 8-12 mg.
Reflexes: Frog, lymph -sac, 3 mg.
Curare Action: Frog, lymph-sac, 10 mg. (Chap. 32, IV, 10); immersion of muscle, 0.2
per cent.
M. F. D.: Pigeon, hypodermic, per kg., 54 mg.
Magnesium Chlorid.
M. F. D.: Dog, vein, per kg., 0.223 gm. (2.35 c.c. of M/8). (Joseph and Meltzer,
1909.)
X Magnesium Sulphate (Crystals).
Anesthetic: All animals, hypodermic, per kg., 1.5 to 1.75 gm. (6 to 7 c.c. of 25 per
cent.). (Rabbit, Chap. 39, XV; Frog, 0.8 c.c. of 25 per cent, per gm., Chap.
32, II, 6.)* (Curare Action, immersion of muscle, 5 per cent., Chap. 32, IV, 4.)*
Fatal: All animals, hypodermic, per kg., 2 gm.
Manganese (Calculated as Metal).
M. F. D.: Dogs, hypodermic, per kg., 10 to 13 mg.; Cat, ditto, 6 to 7 mg.; Rabbit,
ditto, 5 to 6 mg.
Mercuric Chlorid.
Gastro-enteritis: Cat or Rabbit, stomach, per kg., 5 mg. (5 c.c. i : 1000) (Chap. 39,
XVIII, 2).*
Nephritis: Dog or Rabbit, hypodermic, 5 to 10 mg. (5 to 10 c.c. of i : 1000) (Chap.
39, XXI; MacNider, 1912).
Methylene-blue.
Excretion: Man, mouth, 0.15 gm. (Chap. 15, VII, A).*
Fatal: Dog, vein, per kg., 0.125 gm. (25 c.c. of 0.5 per cent.); stomach, per kg., over
i gm. (Tanfiljeff, 1907).
y; Morphin Hydrochlorid or Sulphate.
Respiration: Sedative: Rabbit, hypodermic, per kg., 0.5 mg. (? c.c. of i : 1000)
(Chap. 41, I, i).* Cat, hypodermic, per kg., 2.5 mg.
Maximal: Rabbit, hypodermic, per kg., 2.5 to 10 mg.
Toxic: Rabbit, hypodermic, per kg., 40 mg. (i c.c. of 4 per cent.) (Chap. 41, 1, 3).*
Temperature: Rabbit, hypodermic, per kg., 100 mg. (2.5 c.c. of 4 per cent.) (Chap. 39,
II).* Dog, hypodermic, per kg., 10 to 150 mg.
Gastric Spasm: Dog, hypodermic, per kg., 6 to 7 mg.; Cat, ditto, 8 mg.
Antemetic: Dog, hypodermic, per kg., 10 mg. (£ c.c. of 4 per cent.) (Chap. 38, XVII).*
Excitement: Cat, hypodermic, per kg., 40 mg.
Constipation: Cat, hypodermic, per kg., 40 mg. (milk diarrhea). Rabbit, hypodermic,
per kg., 20 mg. (salt crystal to intestine; reappears with 60 mg.).
Glycosuria: Rabbit, hypodermic, per kg., 50 to 100 mg. (Chap. 39, VIII).*
Narcotic and Preliminary Anesthetic: Dog, hypodermic, per kg., 10 to 20 mg. (J. to
£ c.c. of 4 per cent.) (Chap. 39, XIII; 41, TN).* Cat, hypodermic, per kg., 20 to
60 mg. (£ to i£ c.c. of 4 per cent.) (Chap. 39, XIII; 4i,.TN).* Rabbit, hypo-
dermic, per kg., 5 to 20 mg. (J to % c.c. of 4 per cent.) (Chap. 41, TN).*
Mouse Test: Ordinary: White Mouse, hypodermic, per 15 to 20 gm., 0.05 mg. (0.5 c.c.
of i : looo) (Chap. 39, XIII, 4)-*
Minimal: While Mouse, hypodermic, per 15 to 20 gm., o.oi mg.
Reflexes: Frog, 10 mg. (£ c.c. of 4 per cent.) (Chap. 32, III, 4).*
Narcotic, Tetanic: Frog, 50 mg. (ij c.c. of 4 per cent.) (Chap. 32, II, i).*
M. F. D.: Dog, vein or hypodermic, per kg., 0.4 gm. (Lentharz). Cat, hypodermic,
per kg., 0.04 to 0.08 gm. (G. H. Mueller, 1908). Rabbit, hypodermic, per kg.,
0.2 to 0.32 gm. (Stockman, 1891; Joffroy and Lervaux): stomach, per kg., 0.7 to
i gm. Guinea-pig, hypodermic, per kg., 0.7 gm. White Rat, hypodermic, per
gm., 0.42 mg. (Hunt, 1907). White Mouse, hypodermic, per gm., 0.6 mg.
(Hale, 1910).
APPENDIX H DOSES FOR ANIMALS 331
* Morphin-atropin Anesthesir..
Cat, hypodermic, per kg., Morphin, 20 mg. (J c.c. of 4 per cent.), with Atropin, i mg.
(i c.c. of i : looo) (Chap. 39, XIII, 7)-*
Morphin-atropin-urethane Anesthesia.
Cat, rectal or stomach, per kg., 3 c.c. of the M. A. U. Mixture (I'c.c. «« 10 mg. of
Morphin, 0.2 mg. of Atropin, and 0.2 gm. of Urethane) (Chap. 41, TN).*
Morphin-scopolamin Anesthesia.
Dog or Rabbit, hypodermic. per kg., Morphin, 10 mg. (J c.c. of 4 per cent.); with
Scopolamin, 0.67 mg. (| c.c. of i : 1000) (Chap. 42, II, 4; V).* (Details, Boyt-
cheff, 1907.)
Cat, hypodermic, per kg., Morphin, 2° mg. (J c.c. of 4 per cent.), with Scopolamin,
0.5 mg. (i c.c. of i : looo) (Chap. 39, XIII, 7).*
Muscarin Sulphate.
Cardiac: Dog, hypodermic, per kg., 2 mg.
Toxic: Cat, hypodermic, per kg., \ to | mg.
Bronchial Constriction: Cat, hypodermic, per kg., \ mg.
Fatal: Cat, hypodermic, per kg., i to 2 mg. in two to twelve hours; 3 to 5 mg. in ten
to fifteen minutes.
;s Stimulation: Frog, lymph-sac, 0.5 mg.
M I I ). : /-rot;, lymph-sac, per iogm., 0.12 to 0.23 mg. Toad, lymph-sac, per iogm.,
0.21 to 0.27 mg.
Mustard.
Kmetic: Dog, stomach, teaspoon.
Naphthol, Beta-.
Anthelmintu : Dog, stomach, per kg., 0.06 gm.; Cat, ditto, o.oi gm.
Fatal: Cat, stomach, per kg., o.i gm.
Narcein.
Respiratory sedative: Cat, stomach, o.i gm.
Narcotin.
Light Narcosis: Dog, hypodermic, per kg., 50 mg. Mouse, hypodermic, per 15 to
20 gm., 10 mg.; not fatal (Chap. 39, XIII, 5).* Frog, lymph-sac, 50 to 70 mg.
Fatal: Cat, 3 gm.
Neuronal.
Hypnotic: Dog, stomach, per kg., o.i gm. (Gensler, 1915).
Nickel. Same as Cobalt.
, Nicotin.
Stimulant, Circulation: Mammals, vein, per kg., o.i to 0.5 mg. (^ c.c. to | c.c. of
i : 1000) (Chap. 43, II, 12).*
Emetic: Dog, vein, per kg., 0.35 mg. (Eggleston, 1916) (Chap. 43, II, 12;.*
Stimulant, Respiration: Rabbit, hypodermic, per kg., 0.5 mg. (j c.c. of i : 1000)
(Chap. 41, VI, 4).*
Stimulant, Peristalsis: Rabbit, hypodermic, per kg., lomg. (i c.c. of i : 100) *
34, H).*
Dilation of Ear Vessels: Rabbit, hypodermic, per kg., 10 m^'. (i c.C, of i : 100)
(Chap. 35, 1)-*
Sympathetic I'araly-i-: Rabbits or Cats, vein, |>er kg.. 5 to 10 m^.; local, I per
(both uncertain in dogs) (Chap
Fatal Convulsions: Dog, mouth. : <lrop>. undiluted; Rabbit or ('<;/. ditto, i drop
'[>.*
•C. lymph-sac, i nur. (i C.C. of i : 1000 (Chap. .;-'• IV, 8).*
C- Immersion. /,, per <rnt. (Chap, s . IV, 4).*
M. 1 I) ( . pi • leg., vein, I mg.; hy|M>dermi» . ; mv: . 10 mjr. K<ibbit,
per k«.. vein, 10 mg.; hypodermic. ,*o m>:.; M,,mai h. <o m.
vein. I.2J niK'.: hy|MMlermu. IOB h. <>vcr ^oo mjr. (Hate her and MgRleft-
toll. 1914).
Nitrate, Sodium.
'tit.*
Nitrite, Sodium.
Vascular ,10 to i of 10 JHI 111.
2).* Do-siri. ,,
M.I D Wk \i - ' , , 0.15 mg. (Hale, 1910).
Fatal. Mcthrmo^lohin A',;/./..-/. 1 :(inr<i-pig. ditto,
fng.
Spii >is: Frog, lymph-sai . |xrr gm., 0.55 i
332 APPENDIX
A Nitroglycerin.
Vascular: Mammals, vein, per kg., 0.5 mg. (210 c.c. of i per cent.) (Chap. 43, I, 4).*
Minimal: Rabbit, vein, per kg., 0.05 mg. (Edmunds and Roth, 1908).
/v. Novocain.
M. F. D.: Cat, peritoneum, per kg., 0.45 gm. Dog, peritoneum, per kg., 0.4 gm.;
vein, per kg., 0.2 gm.
Nuclein.
Leukocytosis: Rabbit, hypodermic, 0.5 c.c. of $ per cent, (in eight hours; Coknian,
1007).
Oleate, Sodium.
Hemolysis: Dog, vein, per kg., 10 c.c. of i per cent.
Optochin.
M. F. D.: Frog, lymph-sac, per gm., 0.30 mg. White Mice, ditto, 0.5 mg. (Smith and
Fantus, 1916).
Ouabain (g-Strophanthin).
Circulation: Mammals, vein, per kg., 0.05 mg. QV c.c. of i : 1000) (Chap. 44, HI, S)*
M. F. 1).: Cat, vein, per kg., o.i mg. (Chap. 36, III, 5). Dog, vein, per kg., 0.125 to
0.175 m8-; Rabbit, 0.2 mg. (Hatcher, 1912). Frog, lymph-sac, per gm., 0.0005
mg. (Chap. 36, III, i).*
Heart: Frog, lymph-sac, £ c.c. of i : 50,000 (Chap. 36, IV, 2).*
Oxalate, Sodium.
Kidney Deposits: Rabbits, hypodermic, 0.25 gm., fatal in few hours.
M . F.'D.: Guinea-pig, hypodermic, per kg., 0.4 gm. (Chap. 39, XXI). Chicken, ditto,
0.5 gm. Turtle, ditto, 0.26 gm. Frog, ditto, 0.5 gm.
Anticoagulant: Blood, 0.2 to i : 300.
Oxalic Acid.
Fatal: Rabbit, stomach, 2 to 4 gm. (fatal in one-quarter to one-half hour). Guinea-pig,
hypodermic, o.i gm. Frog, lymph-sac, 40 to 80 mg.
Oxydimorphin.
M. F. D.: Dog, vein, per kg., 60 mg. (dissolve in 0.2 per cent. NaOH); hypodermic,
not fatal in any dose.
v Papaverin Hydrochlorid.
Respiration: Cat, hypodermic, per kg., 40 mg.
Narcotic: Cat, hypodermic, per kg., 100 mg. (Chap. 39, XIII, 6).
M. F. D.: Cat, hypodermic, per kg., 128 mg. (G. H. Mueller, 1908).
Paraldehyd.
Anesthesia: Rabbit, stomach, per kg., i gm. (Chap. 41, TN)* (Mansfeld, 1905).
Vasomotor Paralysis: Rabbit, stomach, per kg., 2 gm.
Anesthesia: Foul, rectum, per kg., 2 c.c. (Edmunds and Roth, 1908).
Paraphenylendiamin.
Eye Changes: Dog, hypodermic, 75 mg. per kg. (Troell, 1916).
Pellotin.
Fatal: Mouse, per kg., 68 mg. (Pincussohn, 1907).
Peptone, Witte's.
Temperature: Rabbit or Cat, hypodermic, per kg., i gm. (5 c.c. of 20 per cent.) (Chap.
39, VI).*
Anticoagulant: Mammals, vein, per kg., 0.25 to 0.5 gm. (2.5 to 5 c.c. of 10 per cent.).*
Shock: Mammals, vein, per kg., 0.2 to 0.5 gm. (2 to 5 c.c. of 10 per cent. (Chap. 43,
III, 6).* (Details, Pearce and Eisenbrey, 1910.)
Permanganate, Potassium.
Antidote: Mammals, stomach, per kg., 15 c.c. of i per cent. (Chap. 40, VI, i).*
Gastritis: Rabbit, stomach, per kg., 0.2 gm. Dog, ditto, o.i gm.
Fatal: Rabbit, stomach, per kg., 0.6 gm. Dog, ditto, 0.4 gm.
Peronin.
Respiration: Rabbit, hypodermic, per kg., 15 mg.
Phenacetin. See Acetphenetidin.
Phenol.
Circulation: Mammals, vein, per kg., 30 mg. (3 c.c. of i per cent.) (Chap. 43, IV, 7)*;
stomach, per kg., i gm.* (recovery by lavage).
Convulsions: Frog, lymph- sac, 10 mg. (i c.c. of i per cent.) (Chap. 32, 1, 9).*
M. F. D.: Cat, hypodermic, per kg., 0.09 gm. (as 2.5 percent.). Rabbit, hypodermic
or stomach, per kg., 0.6 gm. Guinea-pig, hypodermic or peritoneum, per kg.,
0.25 to 0.5 gm. White Mouse, hypodermic, per kg., 0.35 to 0.6 gm. Frog,
lymph-sac, per gm., o.i to 0.6 mg. (as 5 per cent.).
Phenolsulphonephthalein.
Excretion: Man, intramuscular, 0.6 mg. (i c.c. of the solution) (Chap. 15, VII, B).*
APPENDIX H DOSES FOR ANIMALS
Phenylhydrazin Hydrochlorid.
Fatal: Rabbit, hy[x>dermic, per kg., 0.14 gm., death in twenty minu
death on second d.
Phlorhizin.
I)iabete-' \Iammals, hvpodermic, ix?r kg. o.^ mg., minimal effect.' o.i«
Pha,^gav
University
:es; c
Toronto
""->- T»I im 1
effect. Rabbit, hypodermic . IH.T _:m., class work (Chap.
7")/i<r \***ir% i\*»r L-cr /•% T irm f n*\t Huncr^rrkiic^ *
Phosphate, Sodium.
Harmle»: /;.•-. vein. IXT kg.. 35 c.c. of 5 per cent, crystals.*
Phosphorus.
Fatty Liver: Mammals, stomach, per kg., i to 20 mg., in oil or mucilage. Frogs,
>tomach, i to 4 mg. (Details. Ahderhalden, 5, 1233.)
Kidney Lesi<- .-.ml Kar>ner, 1914.
X Physostigmin Salts.
Circulation and Intestine: Mammal*. hypodermic, per kg., 0.5 to 2 mg.*
Antidote to Magnesium: Rabbit, vein, |*r kg., i mg. (Joseph and Melt/er. 1009).
Muscular Fibrillation: Rabbit, vein, JHT kg.. 5 mg. (5 c.c. of i : 1000) (Chap. 32,
Antidote to Curare: Rabbit, vein, per kg., 8 mg. (Magnus, 1908).
M F. D.: Dog, hypodermic, per kg., 4 to 5 mg. Cat, ditto, ,; mg. Rabbit, ditto, 3 mg.
'•a- pig, ditto, 5 mg.
Fatal: /-'rot;, lymph-sac, o.^ mg.
Picric Acid.
Rabbit, vein, per kg., 0.15 gm.; hypodermic, per kg., 0.2 gm.
Picrotozin.
Just Convul>ive: Dog, vein, per kg., 0.3 mg.; hypodermic, per kg., 0.75 mg.; stomach,
per kg., 2.25 mg. Guinea-pig, vein, per kg., i mg.; hypodermic, JKT kg., 5 mg.:
stomach, per kg., 50 mg. Rabbit, vein, per kg., 1.5 "mg.; hyrxxlermii . ;
5 mg.; stomach, per kg., 20 mg. (Hatcher and Fggleston. 1014. Jour. Ann
Assoc., 63, 469).
Convulsion^: ( <;/, hypodermic, per kg., i mg. Guinea-pig. hyjx»dermii . per kg.. 5 mg.
. lymph-sac, <> mg. ' 1.5 c.( . of i : 250) (Chap. ,^-\ I. 8).*
Ml 1 » / ' . . hypodermic, per kg., i .5 mg. Guinea-pig, hyjxxlermic, per kg., 16 mg.
Fata! o mg.
< Pilocarpin Hydrochlorid.
Systemic Fife. t> Mammal*, vein. [>er kg., i mg. d\> c.c. of i per cent.) (Chap. 44,
41, 5).* Mammals. hypi>dermic, per kg., 5 mg. (0.5 c.c. of i per cent.)
V, i).*
Emetic Effects: Dog, vein, per kg., 0.7 mg. (Eggleston, 1916).
Bronchial Constriction: Rabbit, vein, per kg., i mg. (i c.c. of i : 1000
VII).*
Peri.-tal-i-: Rabbit, vein, per kg., 3 mg. (3 c.c. of i : 1000) (Chap. 34, 1, 8).*
Pipcridin.
is: Dog, hypodermic, per kg., 20 mg.
Pituitary Solution.
Cin ulation and Trine: Mammah. vein, per kg., o.i c.c. (Chap. 43. II, 4).*
:.il-is: Rabbit, vein, per kg.. 0.5 i .1 . (Chap. 34, 1, 9).*
Potassium Chlorid.
Cin ulation: Mammals, vein, per kg., lomir. u C.C. Ol i penent.i (Chap. 45, V, 3).*
Frog, lymph -ac , i ; mg. (0.3 C.C of 5 per cent.) (Chap. 32, III, 5).
Pyridin.
Rabbit, hypodcrmi. . JKT kg.. .\5 mg.
Almo-t Fatal: Frog, lymph-sti, o.i gm.
Pyrocatechin.
Prcssor: Dog, vein. |K r 1
>ns: Dog, mg.*
Pyrogtllol.
kg., o.: gn kg., 0.125 gm.
> imin Hydrochlorid.
n. mouth, o..' gm.*
1
Leuk
Ml i Mhitf Mi<i. hyp«
0.7 gm. iSmill- :K-rkg.. o;
•Mnith and l-'ant -.
334 APPENDIX
Rhubarb.
Excretion: A/aw, mouth, i c.c. of Fldext. (Chap. 15, XII).*
C'athartic: Dog, stomach, 5 gm.
Ricin (Merck's).
M. F. D.: Rabbit, vein, per kg., 0.03 mg.; hypodermic, per kg., 0.07 mg.
x Salicylate, Sodium.
Excretion: Man, mouth i gm. (Chap. 15, III; also other salicylates).*
Just Emetic: Cat, hypodermic, per kg., below 0.6 gm.
Convulsive: Cat, hypodermic, per kg., 0.9 to i.i gm. Wild Rat, ditto, 0.65 to 0.75 gm.
Rabbit, ditto, 1.14 to 1.6 gm.
Surely Fatal: Cat, hypodermic, per kg., below 0.9 gm. Wild Rat, 0.65 gm. Rabbit,
ditto, 1.6 gm. (Waddell, 1911, Arch. Inter. Med., 8, 748).
M. F. D.: Dog, vein, per kg., i gm. Guinea-pig, hypodermic, per kg., 2 gm. Frog,
lymph-sac, per gm., i mg. (Blanchier, 1879).
Salol.
Excretion: Man, mouth, 0.3 gm. (Chap. 15, III).*
Salts.
Fatal Doses: Guinea-pigs, vein, Amberg and Helmholtz, 1915, Jour. Pharmacol., 6,
595-
Salvarsan.
Tonic: Rabbit, vein, per kg., 6 to 40 mg.
Albuminuria: Rabbit, vein, per kg., 50 mg. Dog, ditto, 25 to 50 mg.
Fatal Enteritis: Rabbit, vein, per kg., 100 mg. Dog, ditto, 50 to 100 mg.
Acute Death: Rabbit, vein, per kg., 200 mg. (Kochmann, 1912, Muench. Med.
Woch., 59, 1 8).
Santonin.
(For injection, dissolve in dilute NaOH, or use Sodium Santoninate.)
Excretion: Man, mouth, 0.05 gm. (Chap. 15, XI).*
Convulsions and Temperature: Rabbit, stomach, per kg., 0.5 gm. (10 c.c. of 5 per
cent.) (Chap. 39, III).*
Convulsions: Dog, hypodermic, per kg., 0.5 gm.
Fatal: Cat, hypodermic, per kg., i gm. Rabbit, ditto, 2.5 gm.
Sapotoxin.
Fatal: Mammals, vein, per kg., i to 2 mg. Cat, hypodermic, per kg., 40 mg.
Scilla.
Emetic: Dog, stomach, per kg., 2 gm.*
Circulation: Mammals, hypodermic, per kg., i to 10 mg.*
M. F. D.: Guinea-pig, hypodermic, per kg., 0.4 gm.* While Rat, hypodermic, per
kg., 20 gm.* Frog, hypodermic, per gm., 0.6 mg. (Chap. 36, III).*
Scillain.
Fatal: Dog, per kg., i mg.
Scopolamin. See Morphin-scopolamin.
Effects: Frog, lymph-sac, 10 mg. (i c.c. of i per cent.) (Chap. 32, II, 6).*
Senega.
Emetic: Dog, stomach, 5 gm. (Chap. 38, XVI).
Senna.
Excretion: Man, mouth, 2 gm. (Chap. 15, XII).
Silicate, Sodium.
M. F. D.: Mammals, stomach, per kg., 1.5 to 2 gm.; vein, ditto, 0.07 to 0.3 gm. Frogy
lymph-sac, 0.025 to °-1 g™-
Silver Nitrate.
Fever: Rabbit, hypodermic, 2 c.c. of 2 per cent.
Solanin.
Fatal: Rabbit, per kg., o.i gm.
Spartein Sulphate.
Systemic Effects: Mammals, vein, per kg., 5 to 25 mg. (£ to 2.5 c.c. of i per cent.)
(Chap. 44, II, 4)-*
M. F. D.: Various animals, hypodermic, per kg., o.i to 0.15 gm. Rabbit, vein, per
kg., 40 to 60 mg.
Squill. See Scilla..
Stovain.
M. F. D.: Rabbit, per kg., hypodermic, 0.18 gm.; peritoneum, 0.03 gm.; vein, 0.025
to 0.05 gm. (Baylac, 1905;.
Strophanthin (Amorphous or Kombe).
(i mg. of Amorphous Strophanthin is about equivalent, in effect, to f mg. of Ouabain
or 20 mg. of Strophanthus.)
APPENDIX H
DOSES FOR ANIMALS
335
Strophanthus.
Pressor: Mammals, vein, per kg., i mg. (ft c.c. of i per cent.) (Chap. 43, I, 13).*
Fatal: Mammals, vein, per kg., 5 mg. (J c.c. of i per cent.) (Chap. 44, 1, 6).*
M 1 D ('•:/. vein, per kg., 3 mg. Frog, lymph-sac, per gm., 0.006 mg. (Chaj>.
Ill . A'.;/ and Rabhit, (iunn, 1913.
^ Strychnin Salts: Usual Experimental Doses.
A • per kg.) .
Respiratory Stimulant: Rabbit, hypodermic, o.? mg. (0.2 c.c. of i : 1000).
Antagonist to Chloral: Cat, hypodermic, per k^., o.i mg. (o.i c.c. of i : 1000), repeated
(Chap. 39, VII).*
Surely Fatal Dose for Antagonism Experiments: Cat, hypodermic, per kg., 0.75 mp.
(| c.c. of i : 1000) (Chap. 40, I)*; stomach, per kg., i mg. (i c.c. of i : 1000)
.[>. 40, II).* Rabbit, stomach, per kg., 6 mg. (6 c.c. of i : 1000)*; hypo-
dermic, per kg., 0.6 mg. (0.6 c.c. of i : 1000).*
Therapeutic Stimulant: Anesthctiztd Mammals, vein, 0.05 mg. (A c.c. of i : 1000)
P.4i,vn,6)>
Toxic Stimulant: Anesthetized Mammals, vein, 0.25 mg. (\ c.c. of i : 1000) (Chap. 41,
VII, 7).*
Tetanic: Anesthetized Mammals, vein, 0.5 mg. (\ c.c. of i : 1000) (Chap. 43, I, 15).*
Vasomotor Depressant: Anesthetized Mammals, vein, i mg. (Chap. 43, 1, 15).*
Reflexes: Frog (leopard), lymph-sac , 0.02 mg. (0.2 c.c. of i : 10,000) (Chap. 32, III, 5).*
Tetanus: Frog (leopard), lymph-sac, 0.25 mg. (J c.c. of i : 1000) (Chap. 32, V, i).*
Strychnin Salts: Effects on Non-anesthetized Animals.
doses are mg. per kg. Anesthetized animals require considerably larger doses,
varying with the anesthesia.)
Stomach.
Rt-ct.il.
Hypodermic.
Vein.
Other channels.
No perceptible effect :
Dog:
Rabbi!:
0.05
0.4
0.05
0.2
Hyperexcitability
hreckhaft"):
Dog:
Cat:
Rabbit:
Guinea -pig:
on:
0.075
O.I
40
0.075
O.I
0.075
0.08
O.2
3-0
0-5
0-075
O.O2
0-3
Intramusi ular, 0.08
Intramuscular, 3.0
Dog:
•M:
<se:
Frog:
0.175
o.i too. -M
0-57
o.i to 0.24
O.2() to O.4
0.615
0-5
0-5
>>it:
nea-pig:
>ard) :
Toad:
0.47
3-0
0.25
0.58
0.25
0.4
0.4
i.o t-
1.6
0.155
10 to 15 pi
K» than t'
Just Fatal:
Dot:
Cat:
nea-pig:
'•'
Rir
°3-9
lea than
f.O
4.24
2.0
0.35 t" 0.7 5
0-75
0.45 to 0.6
4-5 to 4.75
0.78
2.0
0.4
/°-3 *°
I 0.35
o.;'.
Bladder, 5.5
33 APPENDIX
Sulphate, Sodium.
Diuretic: Dog, vein, per kg., 25 c.c. of 2.5 per cent, (dried) (Chap. 46, 1, i).*
Sulphocyanid, Potassium.
M. F. D.: Pigeon, hypodermic, per kg., 0.5 to 0.75 gm.
Sulphocyanid, Sodium.
Xon-toxic: Dog, vein, per kg., 35 c.c. of 1.2 per cent.*
M. F. D-.: Rabbits, vein, per kg., 0.4 to 0.6 gm. (Corper, 1915).
Suprarenal, Dried.
Pressor: Mammals, vein, per kg., 10 c.c. of i per cent.*
Tartrate, Sodium.
M. F. D.: Dog, vein, per kg., 0.02 gm.
Tetra. See Bcta-tetni-hydronaphthylnmin.
Thallium Salts.
Fatal: Dog, per kg., stomach, 0.5 to i gm.; hypodermic, 0.15 gm. Rabbit, per kg.,
stomach, 0.5 gm.; hypodermic or vein, 0.04 to 0.06 gm.
Thebain Nitrate.
Convulsive: Frog, lymph-sac, 10 mg. (i c.c. of i per cent.) (Chap. 32, II, 6).
M. T. I).: Cut, hypodermic, per kg., 8 mg. (G. H. Mueller, 1908). Rabbit, per kg.,
hypodermic, 21 mg.; vein, 5 to 15 mg.
Theobromin-sodium Acetate (Agurin) or Theobromin-sodium Salicylate (Diuretin).
Diuretic: Man, mouth, 2 gm. Mammals, vein, per kg., 20 to 50 mg. (£ to £ c.c. of
10 per cent.) (Chap. 46, II, 4).* Rabbit, stomach, per kg., 0.5 to i gm.*
Theophyllin-sodium Acetate (Theocin).
Diuretic: Mammals, vein, per kg., 10 mg. (i c.c. of i per cent.) (Chap. 46, III, 4).*
M. F. D.: Dog, vein, per kg., o.i gm. Guinea-pig, ditto, 0.2 gm.
Thiosinamin.
Pleural Effusion: Mammals, vein, per kg., 0.13 gm. (Chap. 37, XIII).
Thiosulphate, Sodium ("Hyposulphite").
M. F. D.: Rabbit, hypodermic, per kg., 1.5 to 2 gm.
Thorium Nitrate.
Non-toxic: Dog, stomach, 25 c.c. of 5 per cent.* Rabbit, stomach, per kg., i gm.*
Tobacco.
0.02 gm. is about equivalent to i mg. of nicotin.
Toluylendiamin.
Fatal Hemolysis. Dog, hypodermic, per kg., 40 mg.
Turpentine Oil.
Diuretic: Dog, stomach, i c.c.
Pleural Effusion: Dog, pleura, i c.c.
Fatal: Dog, stomach, 8 to 30 gm.
Tyramin.
Circulation: Mammals, vein, per kg., 2 mg. (0.2 c.c. of i per cent.) (Chap. 43, II, 6.)*
Uranium Salts.
Hydrops: Rabbit, hypodermic, 5 mg. (i c.c. of \ per cent.), daily three days (Chap.
39, XX)* (Fleckseder, 1906).
Nephritis: Dog, hypodermic, 2 to 15 mg. (MacNider, 1912).
M. F. D. (calculated as metal): hypodermic, mg. per kg.: Dog, 1.66; Cat, 0.41; Rabbit,
0.83; Rat, 0.41; Goat, 1.66; Birds, 40 to 44.
Urea.
Non-toxic: Dog, vein, per kg., 35 c.c. of 0.9 per cent, in isotonic NaCl.
Urethane.
Anesthetic: Rabbit, per kg.: stomach, i gm.; rectum, 0.75 gm.; after morphin, stomach,
0.75 gm.; rectum, 0.5 gm. (Chap. 41, TN).* Cat, stomach, 0.75 gm. per kg.
Dog, ditto, 1.5 gm. per kg. Frog, lymph-sac, 0.2 gm. (2 c.c. of 10 per cent.)
(Chap. 32, TN).*
Hepatic Degeneration: Rabbit, rectum, per kg., 0.6 gm.*
Fatal: Rabbit, rectum, per kg., i.o gm.
Urine, Dog's.
Depressor: Mammals, vein, per kg., 3 c.c. (Chap. 44, III, 4).*
Valerian Oil.
Antispasmodic: Rabbit, hypodermic, per kg., 0.5 gm. (prevents convulsions when
given two hours before Ammonium Carbonate, 0.4 gm. per kg., hypodermic).
Veratrin Sulphate. See also Cevadin.
Muscular: Frog, lymph-sac, 0.05 mg. (0.5 c.c. of i : 10,000) (Chap. 32, I, 10; 33, II).*
Immersion, i : 100,000 (Chap. 33, II).*
Convulsions: Rabbit, hypodermic, per kg., 2 mg. (2 c.c. of i : 1000) (Chap. 40, V).*
Gastric Ulcer: Rabbit, stomach, i c.c. of i per cent. (Chap. 39, XVIII, 4).*
APPENDIX H DOSES FOR ANIMALS 337
Veratrum Viridc.
Vagus Center: Dog, vein, per kg., 5 mg. (\ c.c. of i per cent.).*
Convulsive: Frog, lymph-sac, 5 mg. (0.5 c.c. of i per cent.) (Chap. 32, I, 10).
M. F. D.: Guinea-pig, hypodermic, per kg., 45 mg.
Veronal, Sodium.
Circulation: Mammals, vein, per kg., 0.2 gm. ( .' » .» . «'i" 10 J*T vi-nt.. (Chap. 4 III
s).*
M I . D.: hypodermic, gm. per kg.: Cat, 0.3 to 0.35; Rabbit, 0.4; Frog, 1.5 (Roehmer,
1911).
Yohimbin.
Erection: Mammals, hypodermic, per kg., 0.5 m£.
M. F. D.: Mammals, hypodermic, per kg., 0.5 mg.
Zinc (Zinc-sodium Pyrophosphate or Zinc Valerate, Calculated as ZnO).
I\ir.ily>U: /T(\'. : to i: m^r.
M. I . I ).: Rabbit, hyixxk-rmir or vein. IKT k^.. o.oS to 0.09 gm. Dog, vein, IKT ki:.,
0.07 to o.i :? i*m.
Zinc Sulphate.
Krm-t'u : /;<».«;. stonuuh. 50 i .1 . (»t" i \KT cent. Ca/, ditto, 25 c.c. (Chap. 38, XVI).*
I-'alal: Frog, lymph-sac, per gm.. i to ^ mp.
Zygadenus.
M. I . D.: Rabbit, hypodermic, per kg., 0.6 gm.*
INDEX
ABDERHALDEX'S test, i -M
Abdominal organs, exposure, 256
Abelin test (salvarsan), 74
Absinthe convulsions, 237
Absorption of drugs, 211-216
of salts by intestines, 289, 292
i on reflex time, 14 j
<>n taste, 85
1 orator nerves, frogs, 200
mammals, dissection, 280
operation, 250
-timulation, 184
A. C. E. mixture, 247
Acetanilid, excretion, 94
ts, etc., 63
Acetates, excretion, 92
70
arid, on saliva, 207
tests, etc., 70
Acetone and related substances, tests, etc.,
67
Aceto-nitrile test for thyroid, 220
Acetonuria, experimental, 226
A. ctphenetidin, excretion, 94
tests, etc., 63
Acetyl-morphin. See Heroin.
isolation and tests, 56, 57
> 1 -salicylic acid, tests, etc., 64
Acid as flavors, 85
free, tests, 79
fu< -h-in convulsions, 139
intoxication. 226
on gastric sphincters, 162
on retlcx irritation, 234
on respiration and blood-pressure, 254
radi« al-. inoriranii . tests, etc., 70-84
At idity, a. tu;il. potential and tot,
•ri< , indi. ators, 222
urine, 92
.'26
tc and aconitin, bio-assay, 10=;
blood-pressure and oncometer, 285
perfused, 191, 192
Langendorff, 188
, 198
taste-assay, 120
tests, 59
I 20, 146
Acoustics, 203
V P. Ifin, tc '
A. u|Hin< ture of heart, 258, 259
' solution.
Administration to frogs, 134
Adrenalin. See Epincphrin.
Adsorbents as antidotes, 238
Adsorption, chemical i hanges, 104
experiments on, 97
Agar, diffusion, 102
Agglutination, in
Agglutinin experiments, no
Aggregation of colloids, 108
Air blast, interruption of, 258
Albumose. See Peptone.
on bronchioles, 208
on temperature, 225
Alcohol, blood-pressure and respiration, 275
caffein antagonism, 230
i irculation time, 278
heart, frog, 197
perfused, 193
Langendorff, 188
turtle, 198, 201
mus« le. !«, 158
myocarditis, 294
reflex time, 142
respiration, 253
symptoms, frog, 141
mammals, 230
tests, etc., 66
treatment of poisoning, 230
Aldehyd reactions, 68
Aleuronat suspension, 211
Aliphatic derivative-, tc-ts, 66
Alkali metals, tests. 77, 78
reserve, 92
Alkalies, caustic, tests, 79
incompatibilities, 48
hiking. 1 10
on aiidosis, 226
i-trii -phii
Alkalinity. at tual. potential, and total, 92
Alkaloids, adsorption, 97
antidotes. 96
estimation, 36
isolation
react i
tests, special, 54
Aloes and aloin. nephritis. 233
tests, 38
in urinr.
Alum and aluminum, tests, etc., 73
Alveola in, 80, 242
Amooceptor
339
340
INDEX
Amebic dysentery, 1 29
Amin bases, preparation, 59
Amino-acids, 127
Amino-nitrogen, estimation, 227
Ammonia on respiration and blood-pressure,
253, 254
on shock, 275
on vagus, 234
Ammonium, convulsions, 140
emesis, 221
respiration and blood-pressure. 255
tests, etc., 77
Ampouls, 45
Amyl alcohol, on blood, in
tests, etc., 67
nitrite. See Xitrite, amyl.
Anal sphincter, dilation, on respiration and
blood-pressure, 254
Analgesic activity, comparison, 141
Analgesics, on inflammation, 210
Anaphylaxis, 209, 210
Anasarca, 296
Anesthesia. See Narcosis.
Anesthesia, accidents, 263
blood-pressure and respiration, 261, 262
bottle, 247
cardiogram, 263
frogs, 135
local, 145-148
on inflammation, 210
rapidity and duration, 260, 261
resuscitation, 264
salted frog, 141
spinal, 249
Anesthesiometer, 259
Anesthetic apparatus, 259
mixtures, evaporation, 99
Anesthetics for operative experiments, 246-
249
on circulation and respiration, 256-265
on frog-heart, 197
Aneurysm, aortic, 294
Anilin and derivatives, tests, etc., 63
Animal boards and holders, 249
work, assignment, 22
Animals, experiments on, 131
needed for exercises, 309
Antemetics, 221, 222
Anthelmintics, 129, 130
Antibodies, in, 112
Anticoagulant solutions, 245
Antidotes, animal experiments, 230, 231,
237-239
test-tube experiments, 95-97
Antigen, no
Antimony, emesis, 221
isolation, 52
tests, etc., 73
Antipyretics, 223, 225
Antipyrin, blood-pressure and oncometer,
2g5.
excretion, 94
heart, turtle, 198
metabolism, 226
temperature, 225
tests, etc., 63
Antiseptics, 121-124
Antithrombin, 125
Aorta, frog, 137
perfusion, 175
Aortic aneurysm, 294
cannulae, 170
compression on blood-pressure, 271
on cardiogram, 276
on heart-rate and respiration, 284
insufficiency, 290
stenosis, 291
Apex preparation, mammalian, 190
Apnea on blood-pressure, 255
Apo-atropin, tests, 58
Apomorphin, emesis, 221
hypnotic, 221
isolation, preparation and tests, 57
muscle, 144
Aqua cinnamomi, 41
Arecolin, preparation, 59
Aromatic derivatives, tests, 62
waters, 41
on taste, 85
Arrhenius' hypothesis, 101
Arsenic, circulation, 235, 276
isolation, 52
nephritis, 233
splanchnic stimulation, 276
symptoms in mammals, 232
tests, etc., 73, 74
Arterial injections, 213
pressure, on kidney perfusion, 176
peripheral, 269
rings, 1 66
Arteries, excised, elasticity, 166
vasomotor reactions, 243
Artery, compression, 270
suture, 270
Arthritis, 224
Artificial circulation scheme, 181, 182
respiration, 256-258
for resuscitation, 265
in strychnin poisoning, 239
Ascaris, 129, 130
Aseptic technic, 222
Ash, determination, 40
Asphyxia on blood-pressure and respiration,
255, 261-264
on cardiogram, 264, 288
on vasomotor center, 278
Aspidium, 130
Aspirin, tests, etc., 64
Assays, alkaloidal, 53
Assignment of experiments and reporters, 22
Astringents, 169
on frog mesentery, 176
taste, 1 20
Atophan, tests, etc., 65
Atoxyl, tests, etc., 74
Atropin, blood-pressure, 283
bronchioles, 208, 209
comparison of dog and rabbit, 219
estimation, isolation, and preparation, 58
heart, frog, 197
Langendorff, 188
turtle, 201
INDEX
341
Atropin, intestines, 161, 162, 164
morphin synergism, 229
on cholin, pressor effect, 273
pupil, 203-205
respiration, 253
>aliva, 206
symptoms, 206, 219
urine flow, 283
uterus, 165
vagus mechanism, frog, 200
turtle, 199
-•motor center, 278
Auricle, frog, isolated, 193
Auricular fibrillation, 295
Autol
Autonomic poisons, 163-167
Azo-dye reaction, 62
BMTI:KI\. cultures and media, 122
in feces, 227
Baehr and Pick method of lung perfusion,
208
Balanced saline solutions, 172
Barium, arteries, 166
blood-pressure, 283
heart, frog, 106
turtle, 201
intestines, 161, 162, 164
perfusion, 178, 179
skeletal muscle, 157
tests, etc., 78
urine flow, 283
uterus, 165
Barley water, 42, 125
Bayliss-Starling, intestinal reflex, 161, 163
Be. kmann apparatus, 106
Belladonna bases, distinction, 58
Bellows, artificial respiration, 258
recording, 240.
Bence- Jones proteinuria, 227
Benzidin reaction, 113
Benzoates and benzoic acid, conversion into
hippuric, 92
tests, etc., 64
Berberin, estimation and tests, 57
Bernard, curare experiment, 143
Betain. te^ts, 59
Betanaphthol. tests, etc., 62
Beta-oxybutyri. a. id .67
Beta-tetrahydronaphthylamin, 224, 225
in rhloral |x>isoninR, 231
Bi'.irl.
Bile.
tines, 222
secretion, 222
-cale, 228
Bio-assay- 1 95
anthelmintics. i ^o
anlij' v, 225
•MO
t-H-195
. 270
perfused frog heart 195
Bio-assays, pituitary, 167
smooth muscle, 166-167
suprarenal, 167, 279
thyroid, 220
Birds, anesthesia, 227, 249
operations, 227
urine secretion, 227
Bismuth, antemetic, 222
mixtures, 44
tests, etc., 74
Bladder cannulae, 170
contractions, 166
Bleeding, rabbits, 1 26
Blood, alkalinity, 92
analysis, 125
coagulation, 125
drawing, 1 1 1
experiments on, 1 1 1
flow, human, 180
thermometry, 269
gases, 80, 227
injection, on blood-pressure and urine,
293. 294
kidney perfusion, 178
plasma, 125
platelets, in
precipitants, 116
•quantity, determination, in, 125
tests for, 113
total in body, in, 125
human, 180
viscosity, 108
Blood-corpuscles, agglutination, 1 1 1
count, no, 126
crenation. 1 1 1
experiments on, no
for molecular concentration, 105, 106
hiking, loo-in
microscopic changes. 1 1 1
osmotic resistance, 106, no
ratio to plasma, no
stroma, no
Blood-pressure, compensator, 280
different arteries, 267
human, 170, 180
interpretation, 265
methods, clinical, 179, 180
. 176
manimaK. ane-thetized, 242-246, 265
non-anesthetized, 265
on heart, 183
on heart -rate, 283, 284
percentile changes, 267
jMMti.m OB
relation t«> respiration. .^7. 283, 284
vari.itions in normal dogs, 267
Blood-serum, 125
Blood-vessels, reactions, 166
Blue -print-.
Boards, animal. 240
Body fluids. ..ill.-
--jx.int determination, 40, 53, 107
, 79
raeger's reaction, 38
Br.iin. . ir. illation. 270
compression, 272
342
INDEX
Brain, lipoids, preparation, 71
operations on, 236
perfusion, 171
removal, frogs and pigeons, 135
volume, 270
Brodie operating table, 249
Bromid on convulsions, 237
Bromids and bromin tests, etc., 80
Bronchial muscle, 207-209
spirometer method, 252
secretion, 207
spasm, 208, 209
Brucin, tests, 55
Brunner-Pettenkofer reaction, 37
Buchner press, 127
Bulbs for injection, 212
Burnam's test, 69
Butter, artificial colors, 89
Butyric acid, tests, etc., 71
CACODYLIC acid, tests, etc., 74
Caffein, in alcohol poisoning, 230
blood-pressure, 254, 286, 294
cardiogram, 286
circulation-tune, 278
convulsions, frogs, 140
heart, frog, 197
perfused, 191
Langendorff, 187, 188
turtle, 201
in chloral poisoning, 231
kidney perfusion, 178
volume, 294
muscle, skeletal, 148, 153, 155
reflex tune, 142
respiration, 251, 254
rigor, frog, 140
mammals, 287
tests, etc., 55
urine flow, 294
vasomotor center, 278
Cages, metabolism, 227
Calcium, against chemosis, 210
against convulsions, 237
against pleural effusion, 210
against skin irritation, 210
kidney perfusion, 177
magnesium antagonism, 230
on arteries, 166
on heart, frog, 158
perfused, 191
turtle, 201
on muscle, skeletal, 157
tests, etc., 78
Calomel on bile, 1 23
Calomel-iodid, eye, 217
Calorimetry, 224, 227
Camphor, bromid on, 237
blood -pressure, 288
calcium pressure, 237
cardiogram, 288
channel of administration, 236
curare action, 145
estimation, 40
on heart, frog, 197
Camphor on heart, Langendorff, 187
on respiration, 251
symptoms in mammals, 236
Cannabis, bio-assay, 230
symptoms in dogs, 229
Cannulae, 169-171
Cantharides on skin, 119
tests, etc., 6 1
Capillaries, paralysis and permeability, 267
Capsules, 45
Caramel, adsorption, 98
antidote, 238
tests, etc., 89
Carbohydrates, metabolism, 227
tests, etc., 39
Carbolic acid. See Phenol.
Carbon, in lungs, 80
Carbon dioxid, production, 126
tension, 227, 242
test for respiratory excitability, 242, 252
disulphid, tests, etc., 72
in lungs, 80
monoxid, hemoglobin, 113
preparation and tests, 113
symptoms, 215
Carbonate solutions against clotting, 245
Carbonates and carbonic acid, tests, etc.,
80
Cardamom as flavor, 86
Cardiac. See Heart.
depressants, mammals, 284
dilation, 264
lesions, 290-295
massage, 265
tonics, 284
tracings, frogs, 190-197
mammals, 258, 259
turtle, 198-201
Cardial sphincter, 163
Cardiographic tracings, 258, 259
Cardiomyographs, frogs, 196
mammals, 259
Cardioplethysmographs, frcgs, 192
mammals, 259
Carmin, tests, etc., 89
Carmin-nbrin, no
Carotid artery, clamping, heating, and trac-
tion, 272
operation, 250
Carron oil, 44
Cascara, excretion, 94
Casein, 125
emulsion, 44
Cat method of Hatcher for digitaloic's, 105
Catalase, 126
Cataplasma lini, 46
Cataract, 203
Cathartics, doses for man, 120
emodin, excretion, 94
salts, taste, 87
Catheterization of animals, 225
Cats, anesthetics, 248
Celiac ganglion, 267
Cellulose, 39
Central depressants, frogs, 140-142
mammals, 227-231
INDEX
343
Central nervous system, frogs and other
cold-blooded animals, 135
Cerebral circulation, 270
compression, 272
Cerebrosids, preparation, etc., 71
Cerebrospinal fluid, 218
Cevadin. See Veratrin.
blood-pressure, 281-283
cardiogram, 281
kidney volume, 282
59
urine flow, 283
vasomot 1 78
Charcoal adsorption, 97
as antidote, 238
issay, 167
Chemic antidotes on animals, 238
in test-tube, 95-97
rcises, 35
lockers, 298
Chemosis, 203
Chemotaxis, 129
Chicory. dele, tion, 55
Chloral, absorption, 216
anesthetic, 248
blood-pressure and cardiogram, 276
heart, frog, 197
perfused, 193
turtle, 201
in strychnin poisoning, 239
perfusion of kidney, etc., 178, 179
respiration, 252
symptoms, frogs, 141
mammals, 231
tests, etc., 68
rnent, 231
Chlorates, tests, etc., 80
Chloretone, anesthetic, 248
Chlorid, e\< retion, 93
t<-ts, etc., 81
Chloroform anesthesia, 247, 260-264
blood -pressure, 262-264, 286
by vein, 262
t ardiogram, 264, 286
ether an«Mht-Ma, 248
heart
perfused, 193
I ! >rtT, 187
turtle
mu
nan o-i-. frog, 141
nephritis, 233
poisoning, 262
i, 262-264
285
119
on vagus, 234
, 278
•I tests, 40
On- ol.ite. U
110
Cholin. ,.n . ir. ul.iti.m, 273
test-
Chorda tympani experiment, 206
Chromates, nephritis, 233
tests, etc., 74
Chromogen reaction, 58
Chrysophanic acid, 38
Cilia, 158
Ciliary nerves and ganglion, 202, 203
Cinchona alkaloids, tests, etc., 60
Cinchonin, tests, etc., 60
Circulation, artificial schema, 181, 182
frog, microscopic, 175
rate, human, 180
time experiments, 278
Cirrhosis, hepatic, 233
Citrate, excretion, 92
in transfusion, 270
intestine, 164
kidney perfusion, 177
muscle, skeletal. 157
solution, anticoagulant, 245
tests, etc., 70
Clark's solution.
Class reporters, i
Cleavage products, 127
Clonic convulsions, 138
Clotting, prevention. 245
Coagulation of blood, 125
of milk, 125
solutions to prevent, 245
anesthesia, 146, 147
Cocain against na-al retlcx, 260
epinephrin synergism, 147
heart, turtle, 101
in chloral jxnsoning, 231
intravenous anesthesia, 148
muscle, 144
on temperature, 224
pupils, 204, 205
Mit»titute<. ane-thesia, 146, 147
tests, etc., 57
>ymptom>. mammals, 224
test . 58
Cochineal 89
Codein narcosis, frogs, 141
. BtC.. 56
C« Mi-liver oil. bases, test
di-gui-ing of taste, 87
enubfcn
Coefficient, distribution or partition, 99
Coffe. . 55
Col. hi. iii. tMtS, etc., 59
Col. hi< urn. sympt
Collodion, 42
ulcs and membranes. 10;
Colloids, adsorpti
aggregation, prop. • 108
beofption,
on ' 142
\-nth diarrhea. 233
Mthin. li-t
'03
• lards, 40
neters, 40
. 88
344
INDEX
Colors, coal-tar, 88
detection of, 88-90
Compensating device for blood-pressure, 280
Complement, 1 1 j
Conductivity measurement, 107
Congealing point, 40
Congo- red, diffusion, 10^, 104
Coniin, curare action, 145
preparation, 59
Constant pressure, 168
Contraction curve, form, 153
Convulsants, 135
on temperature, 224
symptoms in mammals, 235-237
Convulsions, location and type, frogs, 137
Copaiba, excretion and tests, 94
Copper, astringent, 159
emetic, 221
tests, etc., 74
Cornea, anesthesia, 146
Coronary circulation, 182
obstruction, 295
perfusion, 171
sclerosis, 295
Corrosives, chemic experiments, 115-118
gastro-intestinal, reflexes, 234
Cotarnin, blood-pressure and kidney vol-
ume, 273
uterus, 165
Cranial nerves, operations, 203
Creatin and creatinin estimation, 227
Crenation, in
Creosote and cresols, tests, etc., 62
Cretinism, 220
Croton oil, on skin, 119, 210
Curare action, 142-145
on blood-pressure and heart, 263
on frogs, 142-144
on pupil, 205
on rabbit, 144
on paper, 142
physostigmin antagonism, 144
Curarin, preparation, 142
Curcuma, tests, etc., 89
Current of rest, 149
source, street, 136
Cyan-hemoglobin, 114
Cyanids, anesthesia, 147
kidney perfusion, 178
symptoms, 214
tests, etc., 72
treatment, 238
Cytisin, tests, 59
DECAPITATION of frogs, 135
Decerebration of mammals, 160
on convulsions, 139, 141
Decoctions, 42
Defibrination, to render blood non-coagu
lable, 246
Delirium cordis, 182, 295
Demulcents. See Colloids.
on reflex time, 142
Depressants, 132
central, frogs, 140-142
Depressants, central, mammals, 227-231
protoplasmic, 158
Depressor nerves, dissection, 250, 268
stimulation, influence on vasomotor
drugs and thyroid, 272
Desiccation, 40
Destruction of organic matter, 52
Dextrin, 39
mucilage, 150
Dextrose. See Su^ir.
Diabetes insipidus, 226
Dialysis, 102
Diastase, 127
Didactic course, schedule, 32
Diethylendiamin, tests, etc., 72
Diffusion, 101
coefficient, 102
Digestion experiments, on animals, 161, 222
Digestive fistulae, 161, 222
products and analysis, 127, 222
secretions, collection, 161, 222
tract, operations, 161, 222
Digitalis, arteries, 166
bio-assay, 193-195
blood-pressure and cardiogram, 282, 286
circulation of frog foot, 175
heart, frog, 196, 197, 219
Langendorff, 188
turtle, 199, 219
infusion, 42
perfusion of kidney, etc., 178, 179
principles, isolation and tests, 60, 61
Dilution, effect on taste, 84
Dilutions, calculation, 134
Dionin chemosis, 205
calcium on, 210
isolation and tests, 56
Diphtheria toxin, shock, 275, 276
Dipping bucket, 168
Dispensing, 41
Dissections, operative, 249
Dissociation coefficient, 107
Distribution of drugs, 217
Diuresis, 289-296
Diuretic coefficient, 290
Dogs, anesthetics for, 246-248
feeding, 227
Dosage, accurate, 134
Doses, calculation, 134
for animals, 320
minimum fatal, 134
Drastic purgatives, determination, 38
Dreser spirometer, 241
Drop recorders, 168
Drums, 150
Drying, 40
powders, 123
Duodenum, nerves, 268
Dusting powders, 123
Dyes, adsorption, 97
EAR, innervation, 167, 203
perfusion, 167, 175
Earthy metals, tests, 77, 78
Eck's fistula, 222
INDEX
343
Edema, measurement, 234
pulmonary, .'55
KtTusion, pleural, 210, 211
Egg experiment (osmosis), 102
Elastometer, 234
Mle- triv stimulation, 136
Kle. tnxardiograms, 290
Kle> trodes, 136
Electrolytes on coagulation, 108
rotytk determination of metals, 53
rometers, 149
Electrophysiology, 149
Elixir, 41
as flavor, 86
Emetics, 220-222
in treatment of poisoning, 230
Emodin principles, assay, tests, etc., 38
in urine, 61, 94
Emprosthotonus, 138
Em unification, avoidance, 51
Emulsin, 37
Kmulsion colloids, 108
Emulsions, preparation, 44
Energy, metabolism, 227
Epileptic convulsions, 235, 237
Epinephrin, absorption, 214
artery, 166
astringent, 159
bio-assay, 167, 279
blood-pressure, 270-278, 286, 288, 291
bronchioles, 208
i ardiai dilation, 264
t ardiogram, 276
chemic tests and estimation, 58
circulation time, 278
cocain synergism, 147
frog perfusion, 175
glycosuria, 226
heart, frog, perfused, 191
Langendorff, 187
turtle, 198, 199, 201
in shock, 275
intestines, 164
kidney volume, 273
mesentery, frog, 1 76
nitrite prc-Mire, 271
on skin, 119
perfiiMon, kidney, etc., 178, 179
pulmonary . ir. illation, -88
pupils, 167, 205
respiration, 275
resuscitation, 265
thyn.id relation, 272
urine flow
uterus, 165, 167
vasomotor center, 278
i pressure, 286
\02
• t of < hcmio lockers, 298
for pharmacodynamics, 304
Erepsin, 127
.i«ay, 166, 173
blood- pn . 286, 288
176
kidney volume, 273
Ergot, pulmonary circulation, 288
rooster-comb, 173
uterus, 165, 166
vein pressure, 286 •
Ergo toxin, arteries, 166
Eriodictyon on taste, 86
Eserin. See Physostigmin.
Ether anesthesia, 246, 260-264
insufflation, 263
rectal, 261
vein, 262
> ilia, 158
• irculation and respiration, 261-264
conductivity of nerve, 147
nine, 246
heart, frog, 197
perfused, 193
Langendorff, 188
turtle, 201
laking, no
muscle, 153, 154
narcosis, frogs, 141
seeds, 158
tests, etc., 67
Ethyl carbamate. See Urcthane.
chlorid, anesthesia, 260
freezing, 148
morphin. See Dionin.
Evisceration, 222
ability of muscle and nerve, 155
Excretion, 90-95, 216, 218
Explosive incompatibilities, 46
Extract, estimation, 40
preparation. 42
Exudates and transudates.
Exudative inflammations, 210
Eye, 202-205
movements, 203
FAT in feces, 227
metabolism, 227
tests, etc., 71
Fatigue, human, 155
muscle, 154, 155
Fatty acids, tests, etc., 71
Feces, 227
on blood-pressure, 282
Feeding bulb, 212
of animals for metabolism, 227
ral vessel- n, .\^O, 251
Ferments, 124-126
. l-.l- .Hd as group reagent, 39
Fever, 224, 225
FibrinoKi-n, 125
l-'i-h. experiments on, 227
Fistula*, digestive, 161
Eck's, 222
Flavors, 84-87
Flaxseed poultice, 46
Hui< I extracts, 42
Fluorescein, eye, 217
renal tot
Fluorwcencr
MIL .rid. muscle, 157
tests, etc., 8 1
346
INDEX
Food, colors and preservatives, 88
utilization, 227
Formaldehyd, tests, etc., 68, 69
Formates and formic acid, tests, etc., 70
Freezing, anesthesia, 148
Freezing-point, determination, 106
Fresenius-Babo method, 52
Froehde's reagent, 55
Frog, administration of drugs, 134
anesthesia, 135
aorta, 137
perfusion, 175
behavior, 135
blood-pressure, 176
boards, 135
brain (structure and destruction), 135
central depressants, 140-142
decapitation, 135
foot, anesthesia, 146
circulation, 175
identification by spots, 195
keeping, 134
mesentery, circulation, 176
needles, 136
perfusion, 167, 173
pithing, 135
sciatic nerve, 135
sensory paralysis, 146
skin secretion, 207
species, 134
Fuchsin convulsions, 139
Fuller's earth, 98
Fusel oil, tests, etc., 67
GAG, 212
Gall-bladder, contractions, 166
Gallic acid, tests, etc., 38
Galvanometers, 149, 290
Gas meters, 241
Gas-balance, Waller's, 259
Gases, absorption, 215
analysis and preparation, 80
blood, 227
intestinal, 80
local anesthetics, 147
work with, 114 .
Gastric acidity, 222
content, 127
movements, 161
sphincters, 162, 163
Gastrin, 207
Gastrocnemius, preparation, 137
tracings, 149
Gastro-enteritis, 231-233
Gastro-intestinal tract, weight, 161
Gels, 1 08
General reactions of plant constituents, 35
Genitalia, male, 166
Gentian as flavor, 86
Germination of seeds, 158
Glands, 206, 207
action current, 149
Glass tubing, 170
Glucose. See Sugar.
on blood-pressure and urine, 294
Glucose, sweetness, 84
Glucosids, general reactions and prepara-
tion, 37
special tests, 60
Glycerin, sweetness, 84
tests, etc., 71
Glycerites, 41
Glycophosphates, tests, etc., 81
Glycosuria, 225, 226
Glycuronic acid, 225, 226
Glycyrrhiza as flavor, 86
Goethlin's solution, 172
Goldfish method for digitaloids, 195
Grehant anesthesia, 248
Guaiac reagent, 126
test for blood, 113
Guaiacol, tests, etc., 62
Guinea-pigs, feeding, 227
M. F. D. of cardiac drugs, 195
Gums, reactions, 39
HALE manometer, 270
Hamburger's blood-corpuscles method, 106
Hartung's frog-heart method, 192
Hearing, 203
Heart. See Cardiac.
analysis of effects, 185
chick embryos, 201
delirium, 182
excised, 182-201
experimental surgery, 290
frog, 190-197
exposed, 195-19?
perfusion for bio-assay, 192, 195
tracings, 196, 197
innervation, 183
irregularities, 182
lesions, 290-295
limulus, 182
lung-kidney preparation, 171
mammalian, acupuncture, 258, 259
excised, 186-190
exposure, 258
tracings, 258, 259
methods of study, 184
nerves, frog, 200
mammals, 250, 280
turtle, 199
reflexes, 285
sounds, 285
standstill, causes, 185
stimulation, 285
tonics, bio-assay, 193-195
turtle, 198-201
valves, movements, 290
weight, 285
Heart-rate, control, 280
influence on output, 183, 279
investigation, 185
mammals, 279-284
influence of blood-pressure, 280
Heat-puncture, 224
Heating of carotids, 224, 272
Hedon and Fleig's solution, 172
Hehner's test, 69
INDEX
347
Hematin, 115
Hematocrit, 106
Hematoporphyrin, 115
Hemochromogen .
HemoKl'>t>i". i 1^-115
Hemolysins, 112
Hemolysis, 106, 109-111
Hemorrhage, blood-pressure, 271, 276, 292
cardiogram, 276
control of .
heart-rate and respiration, 284
kidneys and urine, 292, 294
vasomotor center, 278
Hemostatic tissue extract, .^5 1
Hepatic cirrhosis, 233
Herapathite reaction, 60
Heroin, isolation and tests, 56, 57
K'thylenamin. distribution and excre-
tion. o.\ 117, 218
tests, etc., 69, 70
Hippuric acid, formation, 92
tests, etc., 65
isohn's reaction, 38
Hirudin, 246
Histamin, blood-pressure and kidney vol-
ume, 273
bronchi, 208
uterus, 165, 167
wheal, 119
Holders for animals, 249
Hordenin, preparation, 59
Ho well's solution, 172
Huerthle manometer, interpretation, 261
Hunger contractions, 161
{hint's thyroid test, 220
Hydrastin, hydrastinin, and hydrastis: circu-
lation, 273
convulsions, frog, 140
isolation and tests, 57
uterus, 165
Hydrazin, nephritis, 233
Hydrocephalus, 218
Hydro, yanu a« id. See Cyanids.
Hydrogen j>eroxid, tests, etc., 79
sulphid, excretion and symptoms, 216
I, etc., 83
Hydrogen-ion concentration, 92
Hydropericardium, 293
Hyoson. See Scopolnmin.
Hyperpm-a. nvrphin against, 251
•toni< solutions 101
heart, 158
. 164
kidney perfusion, 177
• »55
on blood-pressure and kidneys, 293
Hyrxx hlorites, tests, etc., 81
.
• m respiration.
. tests, etc., 8 1
I : ..lutions, 101. Set- II
18-220
Iml)il>ition. 104, 108
Iminazolylethylamin. See Histamin.
Immiscible solvents, 51
Immunology. 1 1 j
Incompatibility, 46
Indicator method for CO,, 80
Indol, tests, etc., 62
Indophenol reaction, 63
Inductoria, 136
Infants, metabolism, 227
Infections, experimental, 224
Infiltration anesthesia, 148
Infusions, 42
constant velocity, 214
warmed, 213
Infusum digitalis, 42
Injection. See Infusions.
bulbs, 212
precise amounts, 214
Inorganic poisons, isolation, 52
Insects, 130
Insecticides, 130
Inspection of blood-vessels, 269
Insufflation anesthesia and respiration, 258
experiments, 263
Interaction of drugs, 217
Intermediary metabolism, 227
solvents, 99
Interruption of air-blast, 258
Intestinal absorption, 214
osmosis on, 289, 292
antiseptics, 122
Intestines, 156-165
excised, 163-165
perfusion, 179
Intramuscular injections, 213
Intrapericanlial injection. 265
Intra[>eritoneal and intrapleural injections,
213
Intravenous injection, 213, 214
Inulin, 39
Intervertebrates, 130
muscle, 1 66
Invertin, 127
lodin-calomel effusion, 210
eye, 217
excretion, ui. 04, 95
incompatibilities, 47
tests, etc., 82
lodin as antidote, 96
(ompouix1 •!], 91
liberation by saliva, 91
lips. 120
-kin ilisinle. tion, 222
-t.tin, 118
. 82
lion, ipl
:tion, 2O2
IOJ
75
plnsiol,..- no
reflex effects on < in illation .uxl rcspira-
Irrit.i! 120
348
INDEX
Isolation of poisons, 49
Isometric contraction, 155
Isopurpuric acid reaction, 65
Isotonic solutions, 101
JORISSEN test, 69
Jugular vein, operation, 250
KEYS, cut-out, 136
Kidneys, drugs on, 289-296
exposure, 256
function, 93
perfusion, 176
nerves, 268
section, 103
Kobert's reagent, 55
Kretschmer reflex, 234
Kymographs, 150
LABORATORY rooms, 297
Lactic acid on blood-pressure and respira-
tion, 254
tests, etc., 71
Lactose, sweetness, 84
Laking 109-111
Langendorff hearts, 186-189
Lantern slides of curves, 152
Laryngeal irritation, 234
Lassaigne's test, 36
Lavage, gastric, in poisoning, 238
Laxatives on man, 120
Lead on peristalsis, 162
tests, etc., 75
Lecithin emulsion, 44
preparation, estimation, etc., 71
Leech extract, 245
Leeches, 130
Leucin, 127
Leukocytes, 129
Levers, muscle, 149-151
Levulose, estimation} 39
sweetness, 84
Lewaschew-Pick method of defibrination,
246
Lewen-Trendelenburg, frog perfusion, 173
L'Hermite experiment, 103
Liebermann's test, 69
Lifting power of muscle, 155
Ligatures, 171
Light, 129
Light sensation, 203
Lime-water, 41
Liniments, 44
Linseed poultice, 46
Lipase, 127
Lipins, preparation, 71
Lipochrome, 40
Lipoids, preparation, 71
Liquor calcis, 41
Liquors, 41
Lithium, tests, etc., 78
Liver, cirrhosis and fatty degeneration, 233
function tests, 93
Liver, perfusion, 171
Lloyd's reagent, 98
Lobelia (lobelin), curare action, 145
Local anesthesia, 145-148
Localization of actions, 132
Locker equipment, chemic, 298
pharmacodynamic, 304
Locke's solution, 172
on blood-pressure and kidneys, 295
Lumbar puncture, 218
Lung, absorption by, 215
astringents, 159
excretion by, 216
perfusion, 208
Lycopodium suspension, 295
Lymph hearts, frog, 197
Lymph-sac, frogs, 134
MACERATION, 42
Magnesia mixture, 299
Magnesium, absorption, 292
bulb, 245
curare action, 143, 145
intestines, 164
intracerebral anesthetic, 249
local anesthetic, 147
narcosis, frogs, 141
mammals, 230
solution against clotting, 245
tests, etc., 78
Magnesium-Ca antagonism, 230
Magnet, signal, 245
Male fern, 130
genitalia, 166
Malic acid, tests, etc., 71
Manganese, tests, etc., 76
Manometers, 244, 245
Marine animals, experiments, 227
Mariotte bottle, 198
Marking board, 152
Marquis' reagent, 55
Marsh test (arsenic), 52, 74
Mask-Tambour method, 240
M. A. U. anesthetic, 248
Maximal load, 155
Mayer's reagent, 36, 299
Mean blood-pressure, 244
Measurement of solutions, 135
Meconic acid, tests, 56
Medullary circulation, 270
Melting-point determination, 53, 107
Membrane manometers, interpretation, 245,
266
permeability, 103
Mental tests, 228
Mercuric chlorid, nephritis, 233
symptoms in mammals, 232
potassic iodid, 299
Mercury manometers, 244, 245
purification, 245
tests, etc., 76
Metabolism cages, 227
drugs on, 226, 227
nitrogen, 226, 227
of surviving organs, 127
INDIA
349
Metabolism, respiratory, 242
Metals, antidotes, 06
isolation, 53
special tests, 73-77
time limit test, 77
Methemoglobin, 114
Methyl alcohol, tests, etc., 66
salicylate, tests, 64
Methylene-blue, absorption in blood and
lymph, 218
•el ion, 92
for circulation time, 278
Methyl-xanthins, isolation and tests, 55
Meyer method, arteries, 166
MI. D., 134
.ne>the-ia, 220
feeding, 227
keeping and rearing, 220
test for morphin
Micro-organisms, metabolism, 227
Milk.
artificial colors, 89
secretion, 207
on taste, 85
Millon's reagent, 300
Minimum fatal dose, 134
Miosis, 203-205
Mistura cretae co., 43
Mitral ,-tenosis, 290
Mitscherlich apparatus, 50
Mixtures, preparation, 43
Moisture, estimation, 40
Mol, mol-ions, 101
Molecular concentration, determination,
101, 105. 107
weight, determination, 107
micro-determination, 107
Monkeys, anesthetics, 248
Monocellular organisms, 127, 129
Morphin, antemetic, 221, 222
on anesthetics, 260-262
atropin synergism, 229
atropin-urethane anesthetic, 248
on colocynth diarrhea, 233
glycosuria, 226
heart, frog, 197
perfused, 193
188
turtle, 201
isolation, preparation and tests, 55
local anesthesia, 147
mouse test, 229
on temperature, 224
reflex tune, 142
lopb I'toms in frogs, 140
in mammals, 228
tetanus, frogs, i .; i
uterus, 165
. 1 10
Morphin-'-- icsia, 246
Morphin narcosis syncrgism
Morphin-scopob > sthesia, 261, 263
rirism, 229
Moq. rgism, 265
.'36
Motor endings, depression, 142, 145
nerves, 137
paralysis and stimulation, fro^z
M milage, dextrin, 150
Mucilages, 41
Mucous membranes, 117
astringents, 159
irritation, 120
Mucus secretion, o
Murexid reaction, 55
Muscarin, estimation on frog, 200
frog vagus, 200
perfused frog heart, 192
Muscle, chemical coagulation, 118
corrosion, 117
levers, 149-151
skeletal, 148- 1
smooth, 159-167
tracings, 149, 150
Muscle-nerve preparation, 137
Muscular contraction, 148-157
Mustard, emesis, 221
on skin, 119
Mydriasis, 202-205
Myocarditis, 290, 294
NARCOSIS. See Anesthesia.
Narcotic activity comparison, 141
tests, 56
Narcotics, symptoms in mammals, 229
Nasal tambour method, 240
Neck, operations on, 250
Nephelometer, 108
Nephritic poi>on->. :;;, 234
test-meal, 93
Nerve, conduct ivity. 147
experiments on, 140
lipoids. prepar.it ion. 71
osmotic effects, 155
Nerve-fibers paraly-i-. 147
Nerves, destruction oi .
divi>ion and Simulation, 250
Nessler reagent, 77
Neutral prim iple>. isolation. 51
Neutralization of laustic acid and alkali, 96
Nicotin, blood-pressure, 273
curare a. tion.
ganglia and nerve-nl>ers. iervical symj>a-
thetic. KM
hear; 188
turtle. :oi
inte-tine . 164, 273
on respiration. 253
pupil, loi. .'oo
sahv..
.
mammal v
uterus, 165
Nitrates, test?, etc., 82
. 82
Nitrite, .ur.xl. Mood-pressure, 27
350
INDEX
Nitrate, amyl, on coronary obstruction, 295
on kidney volume, 273, 294
on man, 179
Nitrites, arteries, 166
blood-pressure, 270, 278
in saliva, 91
intestines, 162
perfusion, frog, 175
spleen, etc., 179
respiration, 275
turtle heart, 201
tests, etc., 82
Nitrobenzol, tests, etc., 65
Nitrogen, estimation, 227
Lassaigne test, 36
Nitroglycerin. See Xitritcs.
blood-pressure, 271-278
cardiogram, 276
circulation time, 278
heart rate and respiration, 284
on epinephrin pressure, 271
on pulmonary circulation, 287
tests, etc., 82
vasomotor center, 278
Nitrous oxid anesthesia, 247, 260
estimation, 82
Normal saline solutions, 171
frog, 137
Note taking, 19, 131
Xovocain anesthesia, 147
Nuclease, 127
Nuclein metabolism, 227
OBJECTS of laboratory instruction, 17
Observations, 131
Oculomotor nerve and experiment, 202, 203
Odor, 203
Oils, volatile, preparation and properties, 40
Ointments, 45
Oleate, sodium, hemolysis, no
Oleic acid, tests, etc., 71
Oleoresins, 43
Oncometers, 169
Operative technic, 249-251
digestive tract, 161
Ophthalmoscopy, 203
Opisthotonus, 138
Opium alkaloids, isolation and tests, 56, 57
Opsonic index, 129
Optical manometers, 245
Organic acids, excretion as carbonates, 92
matter, destruction, 52
poisons, isolation, 50
Organs, surviving, metabolism, 227
Osmometers, 101, 102
Osmosis, 101
on absorption, 289, 292
Osmotic effects on muscle and nerve, 155
pressure, 101-103
Ouabain, blood-pressure and kidney vol-
ume, 282
heart, frog, 197
turtle, 201
Outflow recorder, 278
Oxalate nephritis, 233
Oxalates, tests, etc., 71
Oxidase, 126
Oxidation velocity, 126
Oxydimorphin, tests, 57
Oxygen, estimation, 80
pressure, 168
Oxy hemoglobin, 113
PAIN, sensation, 203
Pancreas, extirpation and other experiments,.
222
juice, 127, 222
Papain, 127
Papaverin, symptoms in mammals, 229
tests, 56
Para-hydroxy-phenyl-ethylamin. See Ty-
ramin.
Paralydehyd, anesthetic, 248
tests, etc., 68
Partition coefficient, 99
of phosphatids, 71
Pelletierin, tests, 61
Penetration of antiseptics, 1 24
Pengawaher Djambi, 251
Pepsin, 127
Peptone. See Albumose.
against clotting, 246
shock, 275
Percolation, 43
Perfusion, 167-179
ear, 167
fluid, 171, 172
for heart, 184
for metabolism, 168, 227
frog aorta, 137
vessels, 167
heart, frog, 190-195
excised mammalian, 186-190
turtle, 198, 199
kidneys, 173, 176-179
organs, 176
Pericardial injections, 265
pressure, 293
Peristalsis, 160-162
Peritoneal injection, 213
Permanganate as antidote, 96, 238
Permanganates, tests, etc., 76
Permeability of cells, 106
of vessels, 211
Peronin, isolation, 56
Peroxid, tests, 19
Phagocytosis, 129
Pharmaceutic incompatibility, 49
preparations, 41
testing, 53
Pharmacodynamics course, schedule, 21
Pharmacy course, schedule, 20
Phenacetin, tests, etc., 63
Phenol, abstraction by solvents, 99
and oncometer, 285
blood-pressure, 264, 276
burns, treatment, 120
cardiogram, 264, 276
estimation and isolation, 62, 227
turtle heart, 198
INDEX
351
Phenol, tests, 62
Phenolphthalein, tests, etc., 65
Phenolsulphonephthalein, excretion test, 93
Phenyl >alicylat<
Phenyl-cim honinic avid. tots, etc., 65
Phenyl-quinolin carboxylic acid, tests, etc.,
65
Phenylsulphonates, tests, etc., 62
Phlorhi/in. a. clone, 226
Phosphates, intestines, 164
tests, etc., 83
Phosphatids. preparation, etc., 71
Ph.»sph«>ru-. te<ts, etc., 50, 76
total, estimation, 83
Phosphorus nephritis, 233
PhvMolonic standardization. See Bio-assays.
Physostijrmin, arteries, 166
bronchi, 208
heart, frog. 197
LanijendorfF, 188
turtle, 201
intestines, 161
pupil, 204, 205
tots, 58
uterus, 165
vagus, 200
Physostigmin-curare antagonism, 144
Phytosterin, tests, 71
Picric acid, stain, 118
tests, etc., 65
Picronolic acid, 55
Pit ro toxin, convulsions, frogs, 139
tests, 6 i
Pigeons, removal of brain, 135
Pitzmcnts, animal, 40
I'ills, 45
Pilot arpin, blood-pressure and cardiogram,
282
bronchi, 208
heart, 197
inte-tines, 162, 164
pupils, 204, 205
i. 206
symptoms, mammals, 206
• 59
uterus, 165
rtlc, 199
tests, etc., 72
.lion, 59
Pithing, mammals, 269
Pituitary, bio-assay, 167
blood-pressure and can lion ram, 276
162, 164
kidneys, 295
fi 273
uterus, 165, 167
meral reactions, 35
«.*o
.. 125
ptoti
volume, li
ipharcresis. .-70
•lysis, ion
PI t! human, 180
. 169
Pleural cannula, 241
effusion, 210, 211
injection, 213
Pleurisy
Pneumonia, 224
Poisoning, treatment of, on animal-
231, 237-239
Poisons, isolation, 49
Polygraphs, 180
Polypeptids, 127
Portal vein, nerves, 268
Position on blood-pressure.
Potassium, absorption and excretion, 218
blood-pressure, 218, 288
cardiogram, 288
heart, frog, 158, 197
perfused, 191
Langendorff, 187
turtle. IQU, :oi
reflex time, 142
skeletal muscle, 153, 154, 157
tests, etc., 78
Poultices, 46
Powders, disguising of taste, 87
preparation, 44
Pre- and post-ganglionic fibers, distinction,
206
Precipitins, 112
Preservatives in food, 88
Pressure, constant, rhythmic, 168
in cardiac cavities, 290
respiration, 258
sensation, 203
Protein, estimation in serum and urine, 234
hydrolysis, 127
metabolism, 227
poisoning, preparation, 209
Proteins, 40
precipitant-. 116
Proteolytic ferments, 127
Proteoses, 127
Protoplasmic depressants. 158
poisons, 127, 129
Protozoa, 128, i
Psychic influences on blood-pressure, 180
Psycholoiri. tots, 228
Pulmonary artery pressure, 287
circulation, 270
edema, 255
vasomotor nerves, 268
Pulse, 1 80
1'ulse pressure, 244
Pulse rate of mammals, 207
PUIK ture, heat, 224
Pupillary nerves, 202, 203
Pupils drugi on, 202-205
Purin bases, estimation, 227
\\ lori. (ontrol. 162
.dlol, tc>N, etc., 6a
04
Hi. I48
352
INDIA
Quinin on emigration of leukocytes, 1 29
on metabolism, 226
on muscle, 144, 148, 153, 155
tests, etc., 59, 60
uterus, 165
Quinin-urea hydrochloric! anesthesia, 147
RABBITS, anesthetics, 248
ear, injection, 214
Radio-activity, 129
Rain worms, 130
Raoult's law, 101
Rare elements, tests, etc., 77
Rat- food, and growth, 227
Reagent lists for chemic exercises, 299-304
for pharmacodynamic exercises, 305-
3i9
Recording devices for plethysmograph, 180
Rectal administration, 212
Reduced hemoglobin, 113
Reference books, list, 19
Reflex effects of irritants, 234, 235
stimulation for resuscitation, 264
time, 137, 142
Reflexes on respiration, 252
Refractometers, 108
Registration, photographic, 149
principles, 149, 245
Reighaarmethode, 146
Reinsch's test (metals), 73
Relation of laboratory and didactic instruc-
tion, 1 8
Renal nerves, 268. See also Kidneys.
function tests, 93
Rennin, 125
Reporters, assignment for experiments, 22,
131
Resins, properties, 39, 43
Resorcin, tests, etc., 62
Respiration, artificial, 256-258
carbon dioxid test, 252
chamber, 227
excised tissues, 126
experiments, 239-255
on man, 242
insufflation, 258
methods of studying, 230-242
pressure, 258
pumps, 258
relation to blood-pressure, 267
tracings, 239-241
valves, 241, 257
volume, drugs on, 251
Respiratory metabolism, 227, 242
Resuscitation, 264, 265
Rheumatic arthritis, 224
Rhubarb, excretion, 61, 94
estimation and tests, 38
Rhythm, contractions of skeletal muscle, 157
Rhythmic pressure, 168
Ricin, in
Rimini test, 69
Ringer's solution, 172
on perfused frog heart, 191
Rubefaction, 119
Ruminants, excreta, collection, 227
metabolism experiments, 227
Rusch's solution, 172
SACCHARIN, sweetness, 84
tests, etc., 65
Salicyl and derivatives, excretion, 91, 95
incompatibilities, 48
tests, etc., 63
Salicyluric acid, 64
Saline diuresis, kidney perfusion, 178
infusion, cardiac dilatation, 264
excretion of toxic substances, 238
fate, 296
in aortic aneurysm, 295
stenosis, 291
in blood-pressure, 292, 294
in coronary obstruction, 295
in hydropericardium, 293
in myocarditis, 294
in shock, 275
on urine, 292
solutions, 171, 172
Saliva, 127
secretion, 206, 207
Salol, tests, 64
Salt action, heart, 158
intestines, 161, 164
kidney perfusion, 177
muscle and nerve, 155-157
excretion, 83
metabolism, 227
solutions giving same freezing-point as
i per cent. NaCl, 172
Salted frog, anesthesia, 141
Salvarsan, tests, etc., 74
Santonin,' convulsions, 224
excretion, 94
on temperature, 224
on worms, 129
tests, etc., 6 1
Saponin, bio-estimation, no
laking, no, in
muscle, 144
tests, etc., 37, 38
Scarlet red, skin, 2iov
Schedule of courses, 204
Schoenbein reaction (cyanid), 72
Sciatic nerves, frogs, 135
mammals, operation, 251
stimulation on respiration and blood-
pressure, 254
Scopolamin narcosis, frogs, 141
tests, etc., 58
Scopolamin-morphin on anesthesia, 261, 263
synergism, 229
ether, 265
Seasonal variations in frogs and guinea-pigs,
iQ5
Secondary contractions, 154
Secretin, preparation and tests, 127, 207
Seeds, germination, 158
Selective solvents, 98
Semipermeable membranes, 101
Senega, emesis, 221
INDEX
353
Senna, excretion, 94
Sensory paralysis, peripheral, 145-148
Serun .
frog perfusion, 175
hemolysis. 1 1 1
proteins, estimation, 234
uterus, 209
Shock, peptone, :~$
surgical, 276
toxii..
treatment,
Signal, injei tioi.
maiinet. 151.
Silitates. tests, etc., 83
Silk peptone, 127
Silver, astringent, 159
ts, etc., 77
Single shock, 136
Skin, chemical corrosion, 117
irritants, 119
cat. 210
•35
r^ing of drums. 150
Smoking, hunger contractions, 163
th muscle. 159-167
Snakes, central nervous system, 135
Soap-bark, sneezing, 120
Sodium salts. See under the respective
anions.
ft, etc., 78
Solanin, preparation and tests, 58
Solubility, determination, 40, 53
Solution strengths, 135
Solution-affinity, 103
Solutions, freezing-point of i percent. NaCl,
172
needed for pharmacodynamic exercises,
alphabetic, 305-308
by chapters, 310-319
Solvent-, selci live, 98
Sparu-in. blood-pressure and cardiogram,
281, 286
on urine, 291
preparation, 59
i.il senses, 203
\>\-. blood, i is
Speed of kymographs. 150
180
1 80
Spinal anesthesia, 249
operations, 269
section, 269
r<>gs» 135
Spirits, 42
C piperita?. 42
Spiromrtcr, 241
•51
Splarn hnii nerves, dissection, 207
stimulation. MIMM| pressure, 271
vessels, frogs.
Spier- . 222
nerves and vessels, 268
ii'oratii.n. ;
S<|uil>l> assay, 195
38
Stain for tables, 297
:iiKr. \ital. 100
- on skin, 118
Stands for levers, etc., 150
Starch tests, 39
Stas-Otto method, 50
Stellate ganglia, dissection, 280
Stethograph, 241
Stimulants, 132
Stomach, blood-supply, 161
contents, examination, 161
movements, 161
Stomach-tube, 211, 213
Stop-cocks, perfusion, 168
Stovain anesthesia, 147
Straub-Fuehner method, frog heart per-
fusion, 100-193
Strentgh of 2-y current, 136
Stromuhr, 168
Strontium, tests, etc., 78
Strophanthin tests, 60
Strophanthus, blood-pressure, 271
cardiogram, 276, 281, 288
circulation time, 278
febrile heart, 281
in aortic aneurysm, 295
stenosis, 291
in coronary obstruction, 295
in hydropericardium, 293
in myocarditis, 294
on pulmonary circulation, 288
respiration, 284
shock, 275
uterus, 165
vasomotor center, 278
Strychnin, absorption, 214-216
bio-assay, 138
blood-pressure, 254, 273, 276, 288
cardiogram, 276, 288
curare action, 145
frogs, convulsions, 138
heart, frog, 197
perfusion, 191
l.angendorff, 187, 188
turtle, 201
in chloral |x>isoning, 231
kidney volume, 273
on respiration. 253, 254
reflex tim.
symptoni> in mammals, 236
itment of (MUMming, 238, 239
vasomotor .enter. 278
Subcorncal injc. tion-.
Sul>m.i\ill.ir\ irland. 206
SucklitiKs. metalxdism, 227
M and tests, 39, 226
Sulphates, blood-pressure, 291
intestines, 164
against clotting, 245
test
urinr
Sulphids. !r '
Sulphites, t, •
354
INDEX
Sulphonal, tests, etc., 68
Sulphur dioxid, tests, etc., 83
total, estimation, 83
transformation into sulphids, 126
Sulphuric acid, tests, etc., 83
Suprarenal. See Epinephrin.
Suppuration, 210
Surface area, 134, 224
tension, 108
Surviving organs, 127
metabolism, 227
Suspension colloids, 108
Sweat, collection and secretion, 207
Sweetening agents, 84
Swine cages, 227
Switchboard, 136
Sympathetic dissection, mammals, 250
Synergism, morphin-scopolamin or atropin,
229
opium alkaloids, 229
Synovitis, 224
Syphilis, 129, 224
Syphon recorder, 278
Syringes, 214
Syrup on taste, 85
Syrups, 41
Syrupus, 41
Systematic pharmacology, schedule, 32
TABLES for animal and chemic work, 297
Tablets, analysis, 54. See also under indi-
vidual drugs.
Tadpole test for thyroid, 220
Tambours, recording, 240
Tannin as antidote, 96
as astringent, 159
incompatibilities, 49
tests, etc., 38, 39
Tapeworms, 130
Tartar emetic, emesis, 221
tests, etc., 73
Tartrates, tests, etc., 71
Taste, 203
Tea, analysis, 55
antihemolytic action, no
Tellurite, potassium, bacterial indicator, 122
Temperature, drugs on, 223-225
on muscle, 154
on rabbits, 223
sensation, 203
Testing, pharmaceutic, 53
Tetanizing currents, 136
Tetanus, 138
Thalleioquin reaction, 60
Thebain convulsions, frogs, 141
Theobromin, blood-pressure, 292
muscle, 153
tests, etc., 55
urine, 292
Theopyhilln, blood-pressure and kidneys,
294
preparation and tests, 55
Thermometry, 269
Thigh lymph-sac, frog, injection, 196
Thiosulphates, tests, etc., 83
Thoracic duct, operation, 250
Thrombin, 125
Thymol, tests, etc., 62
Thyroid, acetonitrile test, 220
estimation of iodin, 82
experiments on, 220
sensitization of depressor and epinephrin,
272
tadpole test, 220
Thyroidectomy, 220
Time tracing, 151
Tinctures, 42
Tissue cultures, 1 29
juice, 127
Tissues, osmotic changes, 103
Tobacco smoke, frog, 145
Tongue, anesthesia, 146
Tonic convulsions, 138
Toxicity, local, 119
Toxicologic analysis, 49
Trachea operation, 250
Trachea-tambour method, 239, 240
Tracheal cannulae, 170
irritation, 234
muscle, excised, 208
Tracing paper, 150
Tracings, demonstration of, 152
Transfusion, 270
Transplantation of organs, 129
Tread-mill, 227
Trigeminal reflex, 234
Tripolar block, 146
Trommer's test, 37
Tropococain anesthesia, 147
Trypanosomes, 129, 224
Trypsin, 127
Tryptophan, 127
Tubing, glass, 170
Tuerck's reflex time method, 142
Turmeric, tests, etc., 89
Turtle, central nervous system, 135
heart, 198, 199, 201
Tyramin, blood-pressure and kidney volume,
273
uterus, 165
Tyrode's solution, 163, 172
Ty rosin, 127
ULTRAFILTRATION, 102
Ultramicroscope, 108
Unguentum zinci oxid, 45
Uranium hydrops, 233
Urea, estimation, 227
index, 93
intestines, 164
laking, no
Urease, 127, 227
Ureter cannulae, 170
contractions, 166
pressure, 176
Urethane, anesthetic, 248
frog, 135
reflex time, 142
Urinary antiseptics, 122
I Urine, analysis, 227
INDEX
Urine, bird, 227
collection 290
depressor substances, 282
flow, drugs on, 289-296
pigments, 93
preservation, 227
protein i>timati«m.
Urobilin and urobilinogen, 93
id, isolation, 68
Urochrone, 93
Urticaria. 119
Uterus, anaphylaxis, 209
autonomic drugs, 165
serum, 209
Utili/ation of food, 227
U veil is, 203
VAGI section, blood-pressure and cardiogram,
281
Vagus, dissection, frog, 200
mammal, 250
turtle, 199
poisons, 199-201
on frog, 200
on turtle, 199
reflex, 234
stimulation, 183
cardiogram, 281
circulation time, 278
in hydropericardium, 293
in myocarditis, 294
in pulmonary circulation, 287
kidney volume, 282, 293
on aortic aneurysm, 295
stenosis, 291
on blood-pressure, 281, 286, 292, 293
on coronary obstruction, 295
on heart, frog, 200
turtle, 109
urine, 283, 292
vein pressure, 286
Valerates, tests, 70
Valvular lesions, 290
Yan't Hotf's theory, 101
Varnishing of tracings, 152
Vascular reactions, 269
Vasoconstrktioii and vasodilation, analysis,
266
Vasomotor center, destruction, 268, 269
perfusion method, 277
drugs on, 278
drugs on blood-pressure, etc., 265-279
< .xes, 267
Vein. injc. tion, 213, 214
manometer, 269
collision, kidney perfusion, 176
pressure, 269
human, 180
\ \-in-flow, 168, 270
Venous pulse, 180
Ventricle strips, turtle, JQI
Veratrin. ga>tric corrosion, 232
heart, frog, 196
Derfused, 193
Langendorff, 188
turtle, 201
muscle, 149, 153
on skin, 119
symptoms, frogs, 140
mammals, 237
tests, 59
vasomotor center, 278
Veratrum, 281
blood-pressure and cardiogram, 282, 288
oncometer, 285
Veronal, blood-pressure and respiration, 275
tests, etc., 68
Vesication, 119
Vessel-cannulae, 169-171
Vessel-clamps, 270
Vessel suture, 270
Vessels, permeability, 211
Viscosity, 108
Vital staining, 100
Vitamins, isolation, 59
Yitalis1 reaction, 58
Vividiffusion, 238
Volatile oils, properties, etc., 40
poisons, detection, 50
Volume reduction of gases, 80
Vulpian's reaction, 58
WALLER gas balance, 68
Warm perfusion, 168, 188
Water excretion, 93
heart, 158
kidney perfusion, 177
laking, no, in
manometer, 269
metabolism, 226, 227
perfusion, 155
Water-rigor, 15$
Waters, aroma ti
Williams' apparatus l'r»u he.-.rt, 190
Wound antiseptics, 123
Writing points, 150
YEAST, 128
Young's test, 38
I, 222
ointment, 45
Zymase, 128
SURGER.Y
and
ANATOMY
W. B. SAUNDERS COMPANY
WEST WASHINGTON SQUARE PHILADELPHIA
9, HENRIETTA STREET COVENT GARDEN, LONDON
Elsberg's Surgery of Spinal Cord
Surgery of the Spinal Cord. By CHARLES A. ELSBERG, M. D.,
Professor of Clinical Surgery, New York University and Bellevue
Hospital Medical School. Octavo of 330 pages, with 153 illustrations.
Cloth, $5.OO net. Published July, 1916
There is no other book published like this by Dr. Elsberg. It gives you in
clear definite language the diagnosis and treatment of all surgical diseases of the
spinal cord and its membranes, illustrating each operation with original pictures.
Because it goes so thoroughly into symptomatology, diagnosis, and indications for
operation this work appeals as strongly to the general practitioner and neurologist
as to the surgeon.
Cullen on the Umbilicus
Embryology, Anatomy, and Diseases of the Umbilicus. By
THOMAS S. CULLEN, M. B.f Associate 1 'n>i<-sor of Gynec
Hopkins University. Octavo of 680 pages, with 269 illustrations.
Cl< >th, $7.50 net. Publish* M«y. we
In Dr. Cullen' s new work you get chapters on embryology, anaton
in the newborn, hemorrhage, granulation tissue at the umbilicus, umbilical p<
gastric mucosa at the umbilicus, Meckel's diverticulum. im< sts, p*— «
omphalomesenteric duct, prolapsus of the bowel, concretions, abscess, etc.
The editions you find here may not be the latest.
Write us about any books in which you are interested
SAUNDERS' BOOKS ON
Albee's Bone-Graft Surgery
Bone-graft Surgery. By FRED H. ALBEE, M. D., Professor of
Orthopedic Surgery at the New York Post-graduate Medical School.
Octavo of 417 pages, with 329 text-illustrations and 3 colored plates.
Cloth, $6.5O net. Published November, 1915
ORIGINAL
This book presents Dr. Albee's original applied technic for bone-graft work. The suc-
cessful outcome of any procedure to restore the skeletal architecture depends not only upon
a proper operative technic, but in many cases in a greater degree upon the skill with which
the postoperative external fixation dressing is applied and in the convalescent management
of the case. Dr. Albee here gives you his own successful technic and his own methods of
dressing and management, all illustrated with original pictures.
Dr. Albee is a firm believer in the autogenous graft, and in making it he uses the most
improved instruments and tools, all of which are shown you, and their use in actual work.
This is the only book going fully into this important question of bone surgery, a field of sur-
gical endeavor that is attracting pronounced attention over the entire surgical world.
Smithies and Ochsner's
Cancer of the Stomach
Cancer of the Stomach. By FRANK SMITHIES, M. D., Gastro-
enterologist to Augustana Hospital, Chicago. With a chapter on the
Surgical Treatment of Gastric Cancer, by ALBERT J. OCHSNER, M. D.,
Professor of Clinical Surgery, University of Illinois. Octavo of 500
pages, illustrated. Cloth, $5.75 net. Published January, 1916
A STUDY OF 921 CASES
This work gives you the information gleaned from a study of 921 operatively and path
©logically demonstrated cases of gastric cancer.
This new work is the first monograph upon this subject for more than a decade, and
represents some ten years' study of cases at the University Hospital of Ann Arbor, The Mayo
Clinic, and the Augustana Hospital of Chicago. The wonderful advances made within this
time are of the greatest importance to the clinician, the pathologist, and the surgeon. Dr.
Smithies presents these advances in a most practical way. The chapter on Operative Treat-
ment, by Dr. Ochsner, gives you the most approved and successful technic, illustrating the
various operations with original pictures.
SURGERY AND ANATOMY
Hornsby and Schmidt's
The Modern Hospital
The Modern Hospital. Its Inspiration ; Its Construction ; Its
Equipment; Its Mangement By JOHN A HORNSBY, M.D., Secre-
tary, Hospital Section, American Medical Association ; and RICHARD
E. SCHMIDT, Architect. Large octavo of 644 pages, with 207 illus-
trations. Cloth, $7.OO net March. 1913
HOSPITAL EFFICIENCY
" Hornsby and Schmidt " tells you just exactly how to plan, construct, equip,
and manage a hospital in all its departments, giving you every detail. It gi*«s
you exact data regarding heating, ventilating, plumbing, refrigerating, etc. — and
the costs. It tells you how to equip a modern hospital with modern appliances.
It tells you what you need in the operating room, the wards, the private rooms,
the dining room, the kitchen — every division of hospital housekeeping. It gives
you the duties of the directors, the superintendent, the various staffs, their relations
to each other. It tells you all about nurses' training-schools — their management,
curriculum, rules, regulations, etc. It gives you hundreds of valuable points on
the business management of hospitals — large and small.
Howell Wright, Superintendent City Hospital, Cleveland
11 To me the book is invaluable. I have a copy on my desk and scarcely a day nMBfl
but what I consult it and find what I want."
Allen's Local Anesthesia
Local Anesthesia. By CARROLL W. ALLEN, M. D., Instructor in
Clinical Surgery at Tulane University of Louisiana. Octavo of 608
pages, illustrated. Cloth, $6.00 net.
COMPLETE IN EVERY PARTICULAR
This is a complete work on this subject. You get the history of local
anesthesia, a chapter on nerves and sensation, giving particular attention to /a/*
— what it is and its psychic control. Then comes a chapter on osmosis and
diffusion. Each local anesthetic is taken up in detail, giving very special atten-
tion \ncocain and novocain, pointing out the action on thenervo the
value of adrenalin, para sed by cocain anesthesia, control ot
You get Crile's method of administering adrenalin and salt solution, the exact
way to produce the intradermal whenl. to pinch the flesh for the insertion of the
needle — all shown you step by step. You get full discussions of paraneural,
intraneural, and spinal > intravenous and intra-arterial anesthesia, and
Hackenbuck's regional anesthesia by circumferential injections. You get indica-
an article on
prodn sthesia. Then the production of local anesthesia in the various
taken up in detail. Spinal analgesia and epidural injections are
sidered in a monogragh of 45 pages. PubUahtd October. 1014
SAUNDERS* BOOKS ON
The New Keen's Surgery
Surgery: ITS PRINCIPLES AND PRACTICE. Written by 8 1 eminent
specialists. Edited by W. W. KEEN, M. D., LL.D., HON. F.R.C.S., ENG.
AND EDIN., Emeritus Professor of the Principles of Surgery and of
Clinical Surgery at the Jefferson Medical College. Six octavos of 1050
pages each, containing 3100 original illustrations, 157 in colors. Per
volume: Cloth, $7.50 net.
VOLUME VI GIVES YOU THE NEWEST SURGERY
ALL THE ADVANTAGES OF A REVISION AT ONE-FIFTH THE COST
We have issued a Volume VI of " Keen " — the volume of the newest surgery-
In this way you get all the advantages of a complete and thorough revision at but
one-fifth the cost. It makes Keen's Surgery the best, the most up-to-date surgery
on the market.
In this sixth volume you get the newest surgery — both general and special —
from the pens of those same international authorities who have made the success
of Keen's Surgery world-wide. Each man has searched for the new, the really
useful, in his particular field, and he gives it to you here. Here you get the
newest surgery, and fully illustrated. Then, further, you get a complete index to
the entire six volumes, covering 125 pages, but so arranged that reference to it is
extremely easy. If you want the newest surgery, you must turn to the new
" Keen ' ' for it. Volume VI published March, 1913
Keen's War Wounds
Treatment of War Wounds. By W. W. KEEN, M. D., LL.D.,
Major, Medical Reserve Corps, U. S. Army. i2mo of 169 pages, illus-
trated. Cloth, $1-75 net. Published August, 1917
TREATMENTS BEING USED IN FRANCE
This work, based on a report to the National Research Council, reviews
the latest information. It is obtained by direct communication from the war
hospitals in France. It gives the formulas, preparation, application, and results
of Carrel-Dakin's solution, eupad. eusol, and other antiseptics being used with
such marked success. It takes up the removal of foreign bodies, treatment
and prevention of tetanus, gas infection and gas gangrene, head wounds, ab-
dominal wounds, ambrine and No. 7 paraffin for burns. It is an important
book, instructive from cover to cover.
SURGERY AND ANATOMY
Crandon and Ehrenfried's
Surgical After-treatment
Surgical After-treatment. A Manual of the Conduct of Surgical
Convalescence. By L. R. G. CRANDON, M. D., Assistant in Surgery,
and ALBERT EHRENFRIED, M. D., Assistant in Anatomy, Harvard Medi-
cal School. Octavo of 831 pages, with 265 original illustrations.
Cloth, S6.OO net. Published May, 1912
SECOND EDITION— PRACTICALLY REWRITTEN
This work tells how best to manage all problems and emergencies of surgical
convalescence from recovery-room to discharge. It gives all the details com-
pletely, definitely, yet concisely, and does not refer the reader to some other
work perhaps not then available. The post-operative conduct of all operations
is given, arranged alphabetically by regions. ' A special feature is the elaborate
chapter on Vaccine Therapy, Immunization by Inoculation and Specific Sfra,
by Dr. George P. Sanborn, a disciple of Sir A. K. Wright. The text is illustrated.
The Therapeutic Gazette
"The book is one which can be read with much profit by the active surgeon and will be
generally commended by him."
Papers from the Mayo Clinic
Collected Papers of the Mayo Clinic. K\ \Vn i i AM 1 M \
CHARLKS II. MAYO, M. D., and their ASN • •> Clinic,
Roci ipers of 1905-1909, 1910, 1911, 1912, 1913. Each
an octavo of about 800 pages, illi; volume: Cloth, $5.50
net. 1916 Papers (June, loir,: Cloth, $6.50 net.
THE NEWEST SURGICAL METHODS
These volumes give you all the clinical tea- U the important papers of
W. J. and < . II. Mayo and their associates at The Mayo I'lin:
the advances in operative terhnir. in methods of diagnosis as developed at
great clinic. This new volume, although called the 1916 volumt. v
papers th.it did net appear until «v// into /<?//, quite a few beii ;led for as
late a 1 June. You shoul- volume to your Mayo files.
Bulletin Medical and Chirurtfical Faculty of
" Much of the work done at the M.iyo Clinii
making to character. • • • Represents a most substantial block of
of the work done at the Mayo Clinic and recorded in these papers has been epoch-
modern surgical progrov
A Collection of Papers (published previous to 1909). By
WILLIAM). " I! M \v M. D. Two octavos
of 525 pages each, illustrated. Per set : Cloth, $10.00 net
SAUNDERS' BOOKS ON
Moorhead's
Traumatic Surgery
Traumatic Surgery. By JOHN J. MOORHEAD, M. D., Associate
Professor of Surgery, New York Post-Graduate Medical School and
Hospital. Octavo of 760 pages, with 520 original line-drawings.
Published February, 1917. Cloth, $7.OO net.
FOUR PRINTINGS IN TEN MONTHS
Here is a new book on just this side of your practice — a work for the general
practitioner, the surgeon, the mining, railroad and industrial physician, those
having to do with Compensation Law, accident insurance and claims, and legal
medicine. To those medical men engaged in or preparing for military service
this work is proving of great value. For instance, it gives you at first hand
the open air and sunlight treatment of wounds and Dakin's solution, its formula
and application — treatments the European War has brought forward so em-
phatically.
DaCosta's Modern Surgery
Modern Surgery — GENERAL AND OPERATIVE. By JOHN CHALMERS
DACOSTA, M. D., Samuel D. Gross Professor of Surgery, Jefferson
Medical College, Philadelphia. Octavo of 1515 pages, with 1085 illus-
trations. April, i9i4 Cloth, $6.oo net.
SEVENTH EDITION
A surgery, to be of the maximum value, must be up to date, must be com-
plete, must have behind its statements the sure authority of experience, must be so
arranged that it can be consulted quickly ; in a word, it must be practical and
dependable. Such a surgery is DaCosta' s. Always an excellent work, for this
edition it has been very materially improved by the addition of new matter to the
extent of over 250 pages and by a most thorough revision of the old matter.
Many old cuts have been replaced by new ones, and nearly 1 50 additional illus-
trations have been added.
Rudolph Matas, M. D., Professor of Surgery, Tulane University of Louisiana.
" This edition is destined to rank as high as its predecessors, which have placed the learned
author in the fore of text-book writers. The more I scrutinize its pages the more I admire the
marvelous capacity of the author to compress so much knowledge vn so small a space."
SURGERY AND ANATOMY
Seudder's
Treatment of Fractures
WITH NOTES ON DISLOCATIONS
The Treatment of Fractures: with Notes on a few Common
Dislocations. By CHARLES L. SCUDDER, M. D., Assistant Professor of
Surgery at Harvard Medical School. Octavo of 734 pages, with 1057
original illustrations. Polished Buckram, $6.00 net.
Published June. 1915
THE NEW (8th) EDITION. ENLARGED
WITH 1057 ILLUSTRATIONS
The fact that this work has attained an eighth edition indicates its practical
value. In this edition Dr. Scudder has made numerous additions throughout
the text, and has added many new illustrations, greatly enhancing the value of
the work. In every way this new edition reflects the very latest advances in the
treatment of fractures.
J. F. Binnie, M.D., University of Kansas
" Scudder's Fractures is the most successful book on the subject that has ever been pub.
lished. I keep it at hand regularly."
Scudder's Tumors of the Jaws
Tumors of the Jaws. By CHARLES L. SCUDDER, M. D.t Assistant
Professor of Surgery at Harvard Medical School. Octavo of 395 pages,
with 353 illu , 6 in colors. Cloth, $6.50 int.
Published February, 1912
WITH NEW ILLUSTRATIONS
ftr. Scudder in this book tells you how to determine in each case the./brmr of
new growth present and then points out the host treatment. As the tendency of
malignant disease of the jaws is to grow into the accessory sinuses and toward
the base of the skull, an intimate knowledge of the anatomy of these sinuses is
essential. Dr. Scudder has included, therefore, sufficient anatomy and a number
of illustrations of an anatomic nature. Whether general practitioner or
you need this new book because it gives you just the information you want
SAUNDEKS* BOOKS ON
Cotton's
Dislocations and Joint Fractures
Dislocations and Joint Fractures. By FREDERIC JAY COTTON, M. D-,
First Assistant Surgeon to the Boston City Hospital. Octavo of 654
pages, with 1201 original illustrations. Cloth, $6.00 net.
Published July, 1910
TWO PRINTINGS IN EIGHT MONTHS
Dr. Cotton's clinical and teaching experience in this field has especially fitted
him to write a practical work on this subject. He has written a book clear and
definite in style, systematic in presentation, and accurate in statement. The
illustrations possess the feature of showing just those points the author wishes to
emphasize. This is made possible because the author is himself the artist.
Boston Medical and Surgical Journal
" The work is delightful, spirited, scholarly, and original, and is not only a book of refer-
ence, but a book for casual reading. It brings the subject up to date, a feat long neglected."
The Surgical Clinics of Chicago
The Surgical Clinics of Chicago. By leading Chicago surgeons.
Issued serially, one octavo of 200 pages, illustrated, every other month
(six volumes a year). Per Clinic Year (February to December): Cloth,
$14.00 net; Paper, $10.00 net.
SURGERY FROM THE CLINICAL SIDE
This new bi-monthly considers all departments of surgery from the clinical
side, giving particular emphasis to differential diagnosis and treatment. It gives
you the actual word for word clinics of 40 great teacher-surgeons of Chicago,
representing all the important hospitals of that great center of post-graduate instruc-
tion. You get the day-in and day-out teachings of these men. You get 'their
tried and proved methods of diagnosis; their operative technic; their plans of man-
agement; the benefit of their years of experience, with a wealth of clinical material
unequalled for variety and quantity. Add to the matter of the books the illustrations
by Tom Jones, and the result is practically applied t absolutely fresh teachings, em-
bodying all the new methods.
SURGERY AND ANA TOMY
Kelly & Noble's Gynecology
and Abdominal Surgery
Oynecology and Abdominal Surgery. Edited by HOWARD A.
KELLY, M.D., Professor of Gynecology in Johns Hopkins University;
and CHARLES P. NOBLE, M.D., formerly Clinical Professor of Gyne-
cology in the Woman's Medical College, Philadelphia. Two imperial
octavo volumes of 950 pages each, containing 880 original illustrations,
some in colors. Per volume: Cloth, $8.00 net.
Volume I published May, 1907; Volume II published June, 1908
WITH 880 ILLUSTRATIONS-TRANSLATED INTO SPANISH
This work possesses a number of valuable features not to be found in any
other publication covering the same fields. It contains a chapter upon the bac-
teriology and one upon the pathology of gynecology, and a large chapter devoted
entirely to msdical gynecology, written especially for the physician engaged in
general practice. Abdominal surgery proper, as distinct from gynecolo^
fully treated, embracing operations upon the stomach, intestines, liver, bile-ducts,
pancreas, spleen, kidneys, ureter, bladder, and peritoneum.
American Journal of Medical Sciences
11 It is needless to say that the work has been thoroughly done ; the names of the author*
and editors would guarantee this, but much may be said in praise of the method of present
and attention may be called to the inclusion of matter not to be found elsewhere.1'
Cushing's Brain Tumors
Tumors of the Nervus Acusticus and the Syndrome of the
Cerebellopontile Angle. By HARVEY CUSHIN... M. D., Surgeon-in-
Bent Hrigliam Hospital, Boston. Octavo of 296 pages,
with 262 illustrations. Cloth, $5.00 i Pabuaii«4 n o*«&b«r. i«i?
A STUDY OF 65 CASES
< 'ushing presents here an exh uly of tumors of the acoustic nerve.
He gives you his own technic, and the results of study and obser\ some
cases — a thorough presentation of the subject, embracing history, analysis
of symptoms, physical exam ii •rphology, histology, and operative technic.
You are . only the surgical aspects, but the . . symptomatic, and
pathologic as well The illustrations are particularly note wort!
io SAUNDERS* BOOKS ON
Moynihan's
Abdominal Operations
Abdominal Operations. By SIR BERKELEY MOYNIHAN, M. S.
(LONDON), F. R. C. S., of Leeds, England. Two octavos, totaling
nearly looo pages, with 385 illustrations. Per set: Cloth, $11.00 net.
Published October, 1914
THIRD EDITION. ENLARGED
This new ( $d ) edition was so thoroughly revised that the work had to be reset from
cover to cover. Over 150 pages of new matter and some 85 new illustrations were added,
making 385 illustrations, 5 of them in colors — really an atlas of abdominal surgery. This
work is a personal record of Moynihan' s operative work. You get his own successful methods
of diagnosis. You get his own technic, in every case fully illustrated with handsome pic-
tures. You get the bacteriology of the stomach and intestines, sterilization and preparation
of patient and operator. You get complications, sequels, and after-care. Then the various
operations are detailed with forceful clearness, discussing first gastric operations, following
with intestinal operations, operations upon the liver, the pancreas, the spleen. Two new
chapters added in this edition are excision of gastric ulcer and complete gastrectomy, giving
the latest developments in these operative measures.
Moynihan's Duodenal Ulcer
Duodenal Ulcer. By SIR BERKELEY MOYNIHAN, M. S. (LONDON), F. R. C. S.,
Leeds, England. Octavo of 486 pages, illustrated. Cloth, $5.00 net.
Published March, 1912
For the practitioner, who first meets with these cases, Mr. Moynihan fixes the diagnosis
with precision, so that the case, if desired, may be referred in the early stage to the sur-
geon. The surgeon finds here the newest and best technic as used by one of the leaders
in this field.
" Easily the best work on the subject ; coming, as it does, from the pen of one of the roas-
ters of surgery of the upper abdomen, it may be accepted as authoritative." — London Lancet.
Moynihan's War Surgery
American Addresses on War Surgery. By SIR BERKELEY MOYNIHAN,
M. S., F. R. C. S., Surgeon General, A. M. S. I2mo of 143 pages. Cloth,
$1-75 net. Published November, 1917
The experiences of this English surgeon, who has been in active service since the very
beginning of the war, are contained in these lectures delivered on his recent visit to
this country. He gives you treatments of war wounds — preparation, technic of use, and
relative values of Carrel- Dakin's antiseptic, Bipp, Wright's physiologic solution, Ruth-
erford Morison's method — repair of injuries to lungs, pleura, knee-joint, and periph-
eral nerves. He gives you here a vital message from the front.
SURGER Y AND ANA TOMY 1 1
Fenger Memorial Volumes
Fenger Memorial Volumes. Edited by LUDVIG HEKTOE*, M. D.,
Rush Medical College, Chicago. Two octavos of 525 pages each. Per
Cloth, $15. 00 net. Published May. 1912
LIMITED EDITION
These handsome volumes consist of all the important papers written by the late
Christian Fenger, for many years professor of surgery at Rush Medical College,
Chicago. Not only the papers published in English are included, but also those
which originally appeared in Danish, German, and French.
The name of Christian Fenger typifies thoroughness, extreme care, deep re-
search, and sound judgment. His contributions to the advancement of the world's
surgical knowledge are indeed as valuable and interesting reading to-day as at
the time of their original publication. They are pregnant with suggestions.
Fenger' s literary prolificacy may be judged from this memorial volume— over
looo pages.
Owen's Treatment of Emergencies
The Treatment of Emergencies, By HUBLEY R. OWEN, M. D.,
Surgeon to the Philadelphia General Hospital. Octavo of 350 pages,
with 249 illustrations. Published June, 1917 Cloth, $2.00 net
METHODS AND PRINCIPLES
Dr. Owen's book gives you not only the actual technic of the procedures,
but also the underlying principles of the treatments, and the reason why a
particular method is advised. You get chapters on fractures of all kinds, con-
tusions, and wounds. Particularly strong is the chapter on gunshot wounds,
which gives the new treatments that the great European War has developed.
You get the principles of hemorrhage, together with its constitutional and local
treatments. You get chapters on sprains, dislocations, burns, sunburn, chilblain,
asphyxiation, convulsions, hysteria, apoplexy, exhaustion, opium poisoning,
uremia, electric shock, bandages, and a complete discussion of the various
methods of artificial respiration, including mechanical devices.
Radasch's Anatomy
Manual of Anatomy. By HENRY E. RADASCII, M. D., Assistant
Professor of Histology and Biology, Jefferson Medical College. Octavo
of 489 pages, with 329 illustrations. Cloth, $3.50 net Published AUCUM, i«i?
i) Radasch's new h.ui. II), .ok is complcic in both text and illustra
Every effort has l> omy both easy and in*
foresting, the many illustrations contributing markedly to this end.
12 SURGERY AND ANATOMY
Bryan's Surgery
Principles of Surgery. By W. A. BRYAN, M. D., Professor of Surgery
and Clinical Surgery at Yanderbilt University, Nashville. Octavo of 677
pages, with 224 original illustrations. Cloth, $4.00 net.
Dr. Bryan here gives you facts, accurately and concisely stated, without which no
modern practitioner can do modern work. He shows you in a most practical way the
relations between surgical pathology and the resultant symptomatology, and points out
the influence such information has on treatment. Published November, 1913
Mumford's Practice of Surgery
The Practice of Surgery. By JAMES G. MUMFORD, M. D., Instructor in
Surgery, Harvard Medical School. Octavo of 1032 pages, with 68 1 illus-
trations. Second Edition published June, 1914. Cloth, #7.00 net.
Fowler's Operating Room Third Edition, Reset
The Operating Room and the Patient. By RUSSELL S. FOWLER, M. D.,
Surgeon to the German Hospital, Brooklyn, New York. Octavo of 611
pages, illustrated. Published March, 1913. Cloth, #3. 50 net.
Whiting's Bandaging
Bandaging. By A. D. WHITING, M. D., Instructor in Surgery at the Uni-
versity of Pennsylvania. I2mo of 151 pages, with 117 illustrations. Cloth,
$1.25 net. Published November, 1915
Nancrede's Essentials of Anatomy Eighth Edition
Essentials of Anatomy, including the Anatomy of the Viscera. By CHAS.
B. NANCREDE, M.D., Professor of Surgery and of Clinical Surgery, University
of Michigan, Ann Arbor. Crown octavo, 430 pages; 154 cuts. With an
Appendix containing over 60 illustrations. Based on Gray's Anatomy.
Published October, 1911. Cloth, $1.2 5 net. In Saunders* Question Compends.
Martin's Essentials of Surgery seventh Edition
Essentials of Surgery. Containing also Venereal Diseases, Surgical Land-
marks and Minor and Operative Surgery, and a complete description, with
illustrations, of the Handkerchief and Roller Bandages. By EDWARD
MARTIN, A. M., M. D., Professor of Clinical Surgery, University of Pennsyl-
vania, etc. Crown octavo, 338 pages, illustrated.
Published 1897. Cloth, $1.25 net. In Saunders Question Compends.
Metheny's Dissection Methods
Dissection Methods and Guides. By DAVID GREGG METHENY, M. D.,
L. R. C P., L. R. C. S. (EoiN.), L. F. P. S. (GLAS.), Associate in Anatomy,
Jefferson Medical College, Philadelphia. Octavo of 131 pages, illustrated.
Published November, 1914 . Cloth, $1.25 net.
SURGERY AND ANA TOMY
Crile and Lower's Anoci-Association
Anoci-Association. By GEORGE W. CRILE, M. D., Professor of Surgery,
and WILLIAM E. LOWER, M. D., Associate Professor of Genito-Urinary Sur-
gery, \Yestern Reserve University. 275 pages, illustrated. Cloth, $3.00 net.
Anoci-association is the new way of anesthetizing. It prevents shock, it robs surgery
of its harshness, it diminishes postoperative mortality, it lessens the likelikood of
nausea, vomiting, gas-pains, backache, nephritis, pneumonia, and other postopera-
omplications. You get anoci-association and blood-pressure and tne technic
of nitrous-oxid-oxygen anesthesia. Published July, i«u
Crile's The Kinetic Drive
The Kinetic Drive: Its Phenomena and Control. By GEORGE \V. CRILE,
M. I)., Professor of Surgery, Western Reserve University, Cleveland. < >
of 71 pages, illustrated. Published May, 1016 Cloth, $2.00 net.
In this book Dr. Crile analyzes the mechanism by which the present-day industrial
and commercial "speeding" is accomplished, and relates it to the speeding due to
other stimuli, such as infections, auto-intoxication, physical injury, etc.
Keen's Addresses and Other Papers
Addresses and Other Papers. Delivered by WILLIAM W. Ki i -\, M. K
LL.D., F. R. C. S. (Hon.), Professor of the Principles of Surgery and of din
ical Surgery, Jefferson Medical College, Philadelphia. Octavo volume of
441 pages, illustrated. Published May, i<x>5 Cloth, $3.75 net
Keen on the Surgery of Typhoid
The Surgical Complications and Sequels of Typhoid Fever. By WM. W.
KEEN. M.D., LL.D., F.R.C.S. (Hon.), Professor of the Principles of Surgery
and of Clinical Surgery, Jefferson Medical College, Philadelphia, etc
Octavo volume of 386 pages, illustrated. Published 1898 Cloth, $3.00 net.
Dannreuther's Minor and Emergency Surgery
Minor and Emergency Surgery. By WALTER T. DANNREUTHER, M. D., Sur-
geon to St. Elizabeth's Hospital and to St. Bartholomew's Clinic, New York
City. 1 2mo of 225 pages, illustrated. Cloth, $i .25 net. Published Oeu. 1011
Bier's Hyperemia second Edition, June. 1900
Bier's H> peremic Treatment in Surgery, Medicine, and the Specialties :
A Manual of its Practical Application. By WILLY MLYKR, M. D., Professor
of Surgery at the New York Post-Graduate Medical School and Hospital ; and
PROF. DR. VICTOR SCHMIEDEN, Assistant to Prof. Bier, University of Berlin,
Germany. Octavo of 280 pages, with original illusti. UK. ns. doth. $3.00 net.
• nd this work to all those who are interested in the treatment of infection*, cither atttt* or
chonic, for it i> the only authoritative treatise we have in the English language."— A/w >W* Sf*t*
Journal of Mtdicint.
Morris* Dawn of the Fourth Era in Surgery
Dawn of the Fourth Era in Surgery and Other Articles. By
I. D., New York Post-Graduate Medical School and
Mtal i 2 mo of 145 pages, illustrated. August, 1010. 11.25 net
14 SAUNDERS* BOOKS
American Illustrated Dictionary The New (9th) Edition
The American Illustrated Medical Dictionary. With tables
of Arteries, Muscles, Nerves, Veins, etc. ; of Bacilli, Bacteria, etc. ;
Eponymic Tables of Diseases, Operations, Stains, Tests, etc. By W. A.
NEWMAN BORLAND, M.D. Large octavo, 1179 pages. Flexible leather,
I5.oo net; with thumb index, $5.50 net. Published October, 1917
Howard A. Kelly, M.D., ProfcssorofGynecology, Johns Hopkins UnivtrsHy, Baltimore.
"Dr. Borland's dictionary is admirable. It is so well gotten up and of such con-
venient size. No errors have been found in my use of it."
Golebiewski and Bailey's Accident Diseases
Atlas and Epitome of Diseases Caused by Accidents. By DR.
ED. GOLEBIEWSKI, of Berlin. Edited, with additions, by PEARCE BAILEY,
M.D. Consulting Neurologist to St. Luke's Hospital, New York City.
With 71 colored figures on 40 plates, 143 text-cuts, and 549 pages of
text. Cloth, $4.00 net. In Saunders' Hand- Atlas Series. Published IQOO
Helferich and Bloodgood on Fractures
Atlas and Epitome of Traumatic Fractures and Dislocations
By PROF. DR. H. HELFERICH, of Greifswald, Prussia. Edited, with ad-
ditions, by JOSEPH C. BLOODGOOD, M. D., Associate in Surgery, Johns
Hopkins University, Baltimore. 216 colored figures on 64 lithographic
plates, 190 text-cuts, and 353 pages of text. Cloth, $3.00 net. In Saun-
ders' Atlas Series. Published June, 1002
American Pocket Dictionary New (ioth) Edition
The American Pocket Medical Dictionary. Edited by W. A.
NEWMAN DORLAND, A. M., M. D., Editor "American Illustrated Med-
ical Dictionary." 707 pages. Full leather, limp, with gold edges, -£1.25
net; with patent thumb index, $1.50 net. Published October, 1917
Zuckerkandl and DaCosta's Surgery |d^
Atlas and Epitome of Operative Surgery. By DR. O. ZUCKER-
KANDL, of Vienna. Edited, with additions, by J. CHALMERS DACOSTA,
M.D., Samuel D. Gross Professor of Surgery, Jefferson Medical Col-
lege, Philadelphia. 40 colored plates, 278 text-cuts, and 410 pages of
text. Cloth, $3.50 net. In Saunders* Atlas Series. Published 1902
I
•
CO
University of Toronto
Library
DO NOT
REMOVE
THE
CARD
FROM
THIS
POCKET
Acme Library Card Pocket
U«der P«t "Rcl. Index File-
Made by LIBRARY BUREAU
Live Music Archive
Librivox Free Audio
Metropolitan Museum
Cleveland Museum of Art
Internet Arcade
Console Living Room
Books to Borrow
Open Library
TV News
Understanding 9/11