Library of the University of Alberta, Edmonton, Alberta

Goldberg, W.D.

An investigation into Alberta grown
alkaloidal plants. 1935.

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AN INVESTIGATION INTO ALBERTA GROWN

ALKALOIDAL PLANTS

W. D. Goldberg
Department of Pharmacy

A THESIS

submitted to the University of Alberta
in partial fulfilment of the requirements for
the degree of

MASTER OF SCIENCE

Alberta

Edmonton,
April ,

1935

tfnefld’iflqea

Digitized by the Internet Archive
in 2017 with funding from
University of Alberta Libraries

https://archive.org/details/investigationint00will_0

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TABLE OF CONTENTS

page

I. Introduction . . . 1

II. Literature review (history) . 2

a. Atropa Belladonna . . 2

b . Hyoscyamus niger . . 5

c . Datura Stramonium . 8

d. Datura Me tel . 10

III. Object of investigation . 11

IY. Methods (a review) . 12

a. B.P. 1914 method . 13

b. B.P. 1932 method . 14

c. The method of Rosenthaler . 15

d. The method of Watkins and Palkin .... 15

e. The method of DeKay and Jordan . 17

f. U.S.P.X. method . 18

Methods (an examination) . 23

a. B.P. 3.914 method . 23

b. B.P. 1932 method . 23

c. The method of Rosenthaler . 28

d. The method of Watkins and Palkin .... 28

e. The method of DeKay and Jordan . 28

f. U.S.P.X. method . . 35

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TABLE OF CONTENTS (Continued)

Page

V. Experimental . . . . . 35

a. The test garden . . . 35

b . The seed . 37

c . The growth . 38

1 . Atropa Belladonna . . . 38

i. First year . . 38

ii . Second year . 39

2. Hyoscyamus niger . 39

i. First year . . . . . 39

ii. Second year . . 40

3 . Datura Stramonium . . . . 41

4. Datura Me tel . 43

d. Results of determinations . . . . 44

1 . Atropa Belladonna . . . 44

2. Hyoscyamus niger . 46

First year . 46

Second year . . 47

3 . Datura Stramonium . . . . 48

4. Datura Me tel . . . 49

VI. Discussion . . 50

a. The plant . 50

b. The results . . 51

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TABLE OF CONTENTS (Continued)

Page

1. Moisture contents and ash values .... 51

2. Alkaloidal contents . 54

3. Volatile bases . 56

c. The methods . 59

VII. Summary and conclusions . . . 61

VIII. Acknowledgments . . 62

IX. Bibliography . 63

-

AN INVESTIGATION INTO ALBERTA GROWN

ALKALOIDAL PLANTS

W. D. Goldberg

I. INTRODUCTION

George P. Koch (14) in his paper "The

medicinal

Cultivation ofAPlants" stated -

"War, with all its horrors, and terrible as
the results may be, does produce some good. It
stimulates production, compels efficiency, and
teaches us to be more self-reliant.”

This statement may well be taken as a reason
for the marked increase, as he ably pointed out, in the
cultivation of various medicinal plants in the United
States immediately following 1914, as a result of being
cut off from sources of supply.

This investigation limited itself to the growth
and examination of some of the more widely used plants of
the solanaceous group when grown under Alberta climatic
and soil conditions.

Much work has been done, elsewhere, on this
group of plants, particularly Atropa Belladonna, in regard
to ideal growing conditions, locale, methods of cultivation,
and the use of various fertilizers. The use of various

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methods of extractions and determinations, in efforts to
obtain the greatest possible alkaloidal yield, have also
been reported.

II. LITERATURE REVIEW

a. Atropa Belladonna

Belladonna leaf consists of the leaves and tops
of Atropa Belladonna, Linn, collected when the plant is
in flower, and dried.

Belladonna, or Deadly Nightshade, is a herbaceous
perennial with a fleshy, creeping root, from which arise
several erect, branching stems, to a height of about three
feet. The leaves occur usually in pairs and are usually
of unequal size, oval, pointed, of a dusky green on the
upper surface and paler beneath. The flowers are large
bel^ -shaped, pendant, and of a purplish color. The fruit
is a rounded berry.

The plant is a native of Central and south
Europe where it is found growing in shady places, along
walls and amidst rubbish. It grows vigorously, however,
under cultivation.

During the past fev<r years, much interest has been
shown in the cultivation of this plant, particularly in the
United States. Results show that a high grade of Belladonna

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can be grown there (Berneman (1), Karr (10,11), Koch (13),
Sievers (24).

Plants cultivated in California were found very
rich in active constituents (U.S. Dispensatory XXI) * .

The yield per acre of stems and leaves was somewhat less
than one ton. The California experiment also showed that
the alkaloidal content of Belladonna stems was equal to
that of the leaves (0.51-0.83%) (28).

Sievers (25) found that no relation existed
between the appearance of the leaf and its percentage
content of alkaloid. Specimens, however, he said, which
are musty or contain too much stem, should be rejected as
weak in alkaloidal content.

Koch (13) studied the germination of seeds of
Atropa Belladonna in parts of the United States and found
that the seeds germinate slowly. However, it was found by
workers at the University of Minnesota that the treatment
of seed with concentrated sulphuric acid for forty-five
seconds and washing repeatedly with distilled water, would
produce uniform germination in about fifteen days. Sievers
(23), however, disputed this result.

Koch (13) points out that Atropa Belladonna
germination is greatly enhanced by planting in a hothouse,
and when the plant has grown to a height of about four and
a half inches, over a period of about three months, it is
set out into the experimental field.

Published by J. B. Lippincott Co., Philadelphia.

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Karr (10), however, showed that the cultivated
plant contained very little more alkaloid than did the
wild. He stated that nitrogenous fertilizers tended to
lower alkaloidal content because of larger leaf surface.

This conclusion was reached by observations and determina¬
tions carried out on plants grown in England. Karr (11)
also found that in whatever latitude Atropa Bell a donna is
grown, it was conclusive that the composition of the soil,
the use of fertilizers and seasonal conditions made for
very small variations. His plants were grown in a chalky
soil .

As to moisture, not many workers indicate the
needs of Atropa Belladonna. Koch (13) found that two-thirds
as much weight of leaves and stems was harvested where
moisture to the extent of one-half the physical optimum of
the soil was applied, as was produced where the conditions
of moisture were at the optimum.

Goris and Deluard (7) cultivated Atropa
Belladonna in places both exposed to the sun and in the
shade and found that the plants exposed to the sun produced
three times as many leaves as those in the shade. The
alkaloidal content, they reported, was also much higher.

This finding agreed very well with the findings of Karr (10).

The B.P. directs that the leaves be gathered at
flowering time and should not contain more than 20 % of
stems. But with, of course, always the resulting alkaloidal
content in mind, Borneman (1) states that the plant can be

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harvested any time in the fall and some time before the
first sign of the leaves turning yellow, as at that time
there is a rapid deterioration of alkaloids. Also regarding
the amount of stem to be collected, most workers appear to
agree that the stems may be used as well as the leaves and
the alkaloidal content still not fall below the B.P.
requirements .

All workers are agreed that the leaves, on being
gathered, should be dried as rapidly as possible. This is
well shown by Borne man (1) and others. Koch (IS) states
that the leaves should be rapidly dried at a temperature of
55°-60° as at this temperature, the dried leaves yielded a
maximum of alkaloidal content.

b. Hyoscyamus niger

Hyoscyamus consists of the dried leaves and
flowering tops of Hyoscyamus niger (linn.) and contains not
less than 0.05% of the alkaloids of Hyoscyamus, calculated
as hyoscyamine .

There are about fifteen species of the genus
Hyoscyamus known. They are found in the Canary Islands,
Europe, Northern Africa and Asia. The plant is found in
north and east sections of the United States, occupying
waste plots, old gardens, cemeteries, etc. It is not a
native of North America, but was introduced from Europe.

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The plant occurs in two forms, annual and biennial.
It has a long, tapering, soft, whitish root resembling
parsley, for which it has been mistaken, with disastrous
results .

In England and on the continent, Europe, it grows
along the roads, around villages, in rubbish and in waste
places. Both varieties were formerly cultivated in England,
but now the biennial is chiefly grown.

Gieger and Hesse (1883) were the first to
demonstrate the existence of an alkaloid in Hyoscyamus niger .
Ladenburg (1880) made the statement that there were two
alkaloids present, and these were subsequently shown to be
crystallizable hyoscyamine, cx7H23°3N and the other,
amorphous hyoscine,

Hyoscyamine is the dominant alkaloid though some
hyoscine and atropine do occur.

Henry (9) finds the following total determination

of Hyoscyamus niger:

leaves 0.045%-0.08%

Roots 0.15 -0.17

Stems 0.06 -0.10

Tops 0.07 -0.10

J. A. Borneman (1), who carried out the cultivation
of various solanaceous plants in Pennsylvania, stated that
Hyoscyamus niger was the most difficult of all to grow. The
specimens obtained were usually beautiful, but the assays
were almost always disappointing and rarely gave an
alkaloidal content in excess of 0.06%. His conclusion was

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that unless it would he possible to bring the assay up to
standard it would not pay to cultivate the plant.

Experiments by Newcomb and Haynes (19) showed
that Hyoscyamus nlger cultivated in Minnesota yielded very
high percentages of total alkaloids * the range being from
0 .096$ to 0 .1561$. They recommended (as they did for
Atropa Belladonna) the treatment of the seed with concen¬
trated sulphuric acid before planting, and they found that
they obtained a fairly uniform germination in from twelve
to fifteen days.

Klan (IB) states that with the growth of the
germinating plant, the quantity of alkaloid in its organs
decreases. He states that the order of alkaloid content
in Hyoscyamus niger (both annual and biennial) is

1. Root

2. Flowering tops

3. Fruits

4. Leaves

5 . Stems

The plant growth is rapid and has a characteristic
fetid odor, which disappears on the drying of the leaf.

The growth is heavy and the leaves are large, though
varying in size, the lower leaves being considerably larger
than those above .

Patu (20) in his growth of the plant found that
mildew had completely covered the radicle leaves of the
first year Hyoscyamus, and found in these leaves a lowered
alkaloidal content.

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As, in referring to A. Belladonna, the writer
finds that workers seem agreed that the stems of Kyoscyamus
niger can be used as well as the leaves at harvest time,
and the official requirements still met. This was the
finding of Koch (15, 16).

c . Datura Stramonium

Stramonium consists of the dried leaves and
flowering tops of Datura Stramonium (Linn.) and Datura
tatula (Linn. ) .

Datura Stramonium (the thornapple) is an annual
plant of rank and vigorous growth, usually about three feet
high, but growing in exceptionally rich soil may attain a
growth of six feet. The root is large, greyish- white , with
numerous rootlets. The stem is round, erect, shiny and with
numerous large spreading branches. The leaves are large,
five or six inches in length, dark green on the upper
surface with a lighter green on the lower surface. The
flowers are large, solitary, with a funnel-like corolla.

The fruit is a large, fleshy, ovoid, 4-valved capsule,
thickly covered with very sharp spines and containing
numerous flattened seeds, all attached to a central placenta.
It opens at the summit at ripening, and on drying the seeds
are easily removed.

It is doubtful to what country Datura Stramonium
first belonged. European botanists have referred it to
North America, while American botanists return it to Europe.

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Some consider it as having originated in South America or
Asia, and it is probable that its original habitat is to
be found somewhere in the east.

It is reported to grow wild and abundantly in
Southern Russia, from the borders of the Black Sea east to
Siberia. In the United States, where it has probably come
by seed on board ship in dirt ballast, it is found in
waste spots, road sides and rank soil. Where the plant
grows abundantly, it may be recognized by its rank odor.

But notwithstanding its wide growth in the wild state, it
is being cultivated in order to obtain a drug of uniform
qual ity .

The American supply of the drug comes mainly from
Europe, chiefly Holland, England and Germany.

Koch (14) in his paper "The Cultivation of
Medicinal Plants” states that Stramonium is a weed in nearly
all parts of the United States, and as a weed it attains
considerable growth but is always better under cultivation.
Ordinarily, after seeding, he says, the plant requires no
special care, other than occasional cultivation.

But Miller and Meader (18) found that cultivation,
particularly on D. Stramonium and D. t a tula, with and with¬
out fertilizer, had increased the alkaloidal output. This
came as a contradiction to the findings of Karr on Belladonna
(10) when he found that the greatest growth was in unusually
dry and sunny seasons (0.68% alkaloids). Also in a patch
with much sun and no fertilizer, he obtained 1.035%
alkaloids .

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10

All parts of the plant are active. The parts,
according to most workers, may be gathered at any time from
the appearance of the flowers to the autumn frost. A.R.L.
Dohme (3) examined Stramonium to determine the value in
alkaloids of the various parts of the plant. He found that,
in general, the fresh parts yielded more than the dried,
and that the order of alkaloidal content in the organs
was as follows: stems, seeds, leaves, root.

Gieger and Hesse (1833) first isolated an
alkaloid which they called daturine, and which later was
found to be identical with hyoscyamine. There is also
present in the drug traces of scopolamine. George P. Koch
(13), during his various researches on the solanaceous
plants in regard to their relative alkaloidal content,
compared the relative values of the stem and leaves of
Datura Stramonium. He states that the whole plant, with or
without the root, can be used without fear that the total
alkaloidal value will fall below B.P. or U.S.P.X. standards.

d. Datura Me tel

The Datura leaves are the leaves of Datura Me tel
and Datura fastuosa which are annual plants occurring chiefly
in India and attain a height of three to four feet. While
the plant Datura Me tel Is indigenous to India, it has been
carried to almost all tropical regions. The leaves are

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slightly toothed or wavy-margined, and the flowers are
trumpet- shaped and white. Except for the leaves, Datura
Me tel very closely resembles Datura fastuosa .

In India, the leaves are used as an equivalent
to Belladonna and Stramonium.

Though not a great deal of work has been done on
Datura Metel, it contains chiefly hyoscine (scopolamine)
(9) with occasionally small amounts of hyoscyamine and
atropine .

The plant, though belonging to the Solanaceous
group of plant drugs, is not official, but forms the chief
source of hyoscine.

The alkaloid is most commonly extracted from the
leaves, which contain 0.25-0.55% of alkaloids, the other
parts of the plant yielding varying amounts of alkaloids
as follows:

Fruits 0.12%

Roots 0.1 - 0.22%

Seeds 0.23- 0.50% (28)

III. OBJECT OF INVESTIGATION

This investigation was prompted by the desire to
know if the plants, normally grown in central and southern
Europe and in southern England, could be successfully grown
in our normal Alberta climate, with its comparatively short
growing season, and to see if these plants would produce
alkaloidal contents, at least equal to the B.P. requirements.

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No added cultivation, other than periodic watering during
the dry season and occasional weeding, would be used.

It was then with a sense of questioning if Alberta
could produce at least some of its own medicinal plants
successfully that this investigation was undertaken.

The work of Newcomb and Haynes (19) mentioned
previously, in connection with solanaceous plants in
Minnesota, acted as an incentive for the investigation,
since the growing conditions of Minnesota and Alberta are
very similar.

IV. METHODS (A review)

Much work has been done on the extraction and
assay of the solanaceous plants, particularly A trope
Belladonna .

Many methods have been evolved, each method
giving the particular worker fairly consistent results,
but the results of the various workers remaining out of
agreement. This may be due to personal error or to
manipulation, as various workers using the same method
obtain varying results. Similarly Sievers (24) showed a
variation of the alkaloidal content of the leaves of the
same plant, ranging from 0.110$ to 0.766$.

The chief point of contention in the actual
method of assay of particularly the solanaceous plants
has been the relative instability of the alkaloids contained

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therein. Recent workers, among them Watkins and Palkin
(26), Dekay (27) and Jordan (2), Evans and Goodrich (5),
Evans and Davy (4) are satisfied that the alkaloids are
stable enough to permit of extraction by a hot ether, or
other solvent, apparatus. Schow and Bjerregaard (21),
whi^e working with the sterilization of various substances,
found that solutions of atropine could be heated at 120°
for twenty minutes without danger of decomposition. Dekay
and Jordan (2) formed the conclusion, based on their
experiments with the alkaloids, that the alkaloids are
much more stable than they are usually assumed to be.

When chloroform solutions of them are evaporated, they can
be heated at water-bath temperature for one or two hours,
without decomposition. Watkins and Palkin (26) found that
the alkaloids heated in acid or alkali on a water-bath
showed no apparent loss. Consequently, many new methods
have been evolved for the extraction of solanaceous plants.

Before going on to the newer methods, established
methods were first examined for their value, keeping always
in mind the fact that the B.P. still considers these
alkaloids too unstable in solution to be extracted by any
means other than the maceration and cold extraction.

a. B.P. 1914 Method

This method directed the maceration of the powder
in #60 powder, directly in the percolator, fitted with a

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14

tap and plugged with cotton. Maceration was allowed for
one hour, then percolation carried on slowly, into a
separator containing N/l sulphuric acid. The final
chi or o formic residue of alkaloids was evaporated and
titrated with 10 mils of 0.05 N sulphuric acid and 0.05 N
sodium hydroxide, using tincture of cochineal as indicator.

b. B.P. 1932 Methods

The new methods for A. Belladonna and D.
Stramonium are identical.

The powder is directed to be of a #60 fineness
and is macerated in a flask for one hour, in contact with
ammonia/ alcohol x and ether. It is transferred to a
percolator plugged with cotton and percolated for not more
than three hours or until total extraction of alkaloids is
effected. To the percolate is added 0,5 N hydrochloric
acid and the alkaloids shaken out in the usual manner. The
final chloroform solution of alkaloids is evaporated to
dryness and dried for half an hour at 100°. The residue
is titrated using 0.02 N sulphuric acid and 0.02 N sodium
hydroxide and methyl-red as indicator.

For Kyoscyamus, which contains a smaller amount
of alkaloids, much the same procedure is used, but since
forty grammes of drug are used, the standard percolator is
used. Maceration and percolation are similar to those used
for Belladonna, but percolation should not require more

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than two hours. Since, however, the alkaloids of Hyoscyamus
are more sensitive to heat than are the others, the mixed
acid solutions of alkaloids are made alkaline and evaporated
in vacuo at a temperature not exceeding 40°. It is then
extracted with chloroform as usual , the chloroform
evaporated off and the residue dried at 80° for two hours.
The residue is titrated using 0.02 N sulphuric acid and
0.02 N sodium hydroxide, and methyl -red as indicator.

c. The Method of Rosenthal er

For the total extraction of the drug plants, the
procedure as outlined by Rosenthaler in his "The Chemical
Investigation of Plants" (1930)* appeared to be the most
thorough, and is a method by which all constituents occurring
in the plant may be isolated and recognized.

d. The Method of Watkins and Palkin (27)

This method was devised particularly for the evalu¬
ation of Hyoscyamus. The alkaloids! yield by this method
has shown results in some cases as much as three times as
great as that obtained by the U.S.P. IX and X methods.

The essential steps in this mechanical extraction are
i. Treatment of the mass of crude drug with an
alkaline medium to liberate the alkaloid. Macerate over night.

* Published by G. Bel! and Sons, Limited, London.

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ii. Continuous extraction by means of the
device described, using ether as a solvent.

iii. Purification process of the alkaloidal
residue, during which chlorophyll, tars and other extract¬
ives are removed.

iv. Extraction of the alkaloids from aqueous
solution by means of another continuous device.

v. Titration of the alkaloids.

The concentration of ammonia in the original
maceration has been the subject of considerable enquiry,
and it may be in this connection that the varying results
occur with different workers.

It is conceivable that too low a concentration
of ammonia will not completely free the alkaloids from the
crude drug, while too great a concentration is not advisable
because of the instability of the alkaloid hyoscyamine .

With this in mind, Watkins and Pal kin extracted
several samples with all conditions being uniform, but
varied the concentration of ammonia.

They found that 7 cc. of 16 N ammonia proved the
most efficient, as may be seen from the following table
taken from their paper.

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TABLE I

Effect of ammonia concentration on yield
of alkaloid

Ammonia

Alkaloid

Quantity
cc .

Normality

Mgm.

Per cent

5

5

19.09

0.159

16*49

0.138

4

16

20.54

0.171

21.12

0.176

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16

20*54

0.171

20.54

0.171

9

16

20.54

0.171

21.12

0.176

These workers used 0*02 N acid and 0,02 N alkali
and used methyl red as an indicator.

e. The Method of DeKay and Jordan (2)

These workers, like Watkins and Palkin, used a
mechanical extraction apparatus in much the same manner as
did Watkins and Palkin, except that they used the regulation
Soxhlet apparatus instead of one specially devised. The
assay was essentially the same, except that these workers
extracted completely the volatile bases and determined

them

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They concluded, from their experiments, that the
alkaloids of Hyoscyamus (the subject of their paper) are more
stable than usually thought. They determined also, that
most procedures extract certain volatile bases which they
determined as triraethyl amine and a primary amine, and the
indication of the presence of dime thyl- amine , along with
the alkaloids.

DeKay and Jordan used 0.02 N acid and 0.02 N
alkali in their titration of the alkaloid, and methyl red
as indicator.

f. Abstract of Proposed Changes to U.S.P.X.

(29) and U.S.P.X. Method

This paper dealt with two new proposed methods
for assaying crude drugs, i.e.,

1. Aliquot part method (by maceration only),

2. Total extraction method (by maceration and
percolation) .

The second procedure more closely resembles the
B.P. method so it only will be considered.

The drug is placed in a small cylindrical
percolator (see Plate I, Fig. 1) previously prepared by
packing the outlet with cotton. Add solvent to completely
saturate the drug, mix, cover the percolator, allow to stand
five minutes and add the ammonia solution and mix again.

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Percolator Types

Fig. 1

Fig. 2

Plate I

20

After macerating for one hour, percolate slowly until
completely extracted. This method uses 0.10 N sulphuric
acid and 0.02 N sodium hydroxide, and cochineal or methyl
red, as indicator.

With the foregoing review, it appears then that
there are two procedures open to the worker, viz.,

1. Extraction by cold method (B.P. and U.S.P.X.),

2. Extraction by continuous or hot method.

It might be well then to examine step by step the basic
principles of a plant assay, with a view to determining
the value or objections to the various methods outlined.

1 . The crude drug.

The drug must be well divided, at least to a #60
powder and must represent the whole drug. It is in¬
sufficient to grind the powder, sift it through a #60
sieve and discard midrib or stem portions which refuse to
go through the sieve. They must be further ground and then
incorporated w ith the previous portion. It is further
important that the fine powder be thoroughly mixed.

It is evident that upon the completeness and
fineness of the sample rests the accuracy of the resulting
determinations .

In this respect, all of the foregoing methods

agree .

2 . The extraction.

Upon the differences of opinion as to the relative
stability of the solanaceous alkaloids rests the fact that

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there are so many varied procedures advanced for the assay
of a plant drug.

All procedures agree upon the maceration, the
hot methods, however, directing maceration over night,
while the cold methods macerate for not more than two
hours. With the proper concentration of ammonia, it is
conceivable that two hours should be sufficient to free all
alkaloids .

The tendency to the production of an emulsion
when a solution containing vegetable extractives is shaken
with an immiscible solvent, forms a great and common
difficulty. This difficulty, however, is now almost
wholly surmounted in most methods by the addition of
alcohol in the first shaking out with acid, instead of
acid alone. Emulsions in the hot method are not very
common. All procedures extract considerable coloring and
other matter in the original extractive, and this often
interferes in the final determination by coming through
in traces. The procedure by DeKay and Jordan (2) and
Watkins and Palkin (27) precipitates this coloring matter
by adding acid. The solution is then filtered and the
chlorophyll left behind.

The alkaloidal residue is not treated similarly in
the various procedures. All methods agree that there is
extracted, along with the alkaloids, an appreciable amount
of volatile principles and direct the heating of the final

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residue at varying temperatures, governed by the relative
stability of the alkaloid.

In this respect, independent workers agree that

the solanaceous alkaloids are more stable than they are

generally reported to be (vide p. 13). This thought is

coming into the methods of the B.P. and U.S.P.X. since the

residues are heated, in the case of Belladonna and

o

Stramonium, at 100 for one-half an hour.

The method outlined by DeKay and Jordan isolates,
weighs and calculates the actual percentage present of
volatile principle, while other methods simply heat the
residue to drive off these volatile principles and then
titrate .

The indicators in all cases suggested are cochineal
or methyl red and the use of one or the other is left to
the discretion of the worker. Self (22) to a certain extent
defends cochineal, though methyl red is preferred. To
obtain high accuracy with either indicator, Self suggests
that the volume of titration liquid be kept as low as possible,
especially in assays where the weight of alkaloid obtained
is very small. This expedient, he says, produces a much
sharper end-point.

Evers (6) suggested that Brom- phenol -blue would
prove the best indicator for titrating the solanaceous
alkaloids, and that methyl red ranked next. The range of pH
of Brom- phenol -blue is 2. 8-4. 6; the range of pH of methyl
red is 4. 2-6. 2. Mellon and Tigelaar (17), however, found

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23

that, though methyl red did not have quite the best pH range
for titration purposes, yet the error involved in the
titration of atropine by methyl red is so small it may be
disregarded.

On looking back over the above review of the
methods, one is struck with the apparent similarity in
procedures in all steps, save that of extraction. Here the
opinion and findings of the individual worker enter to make
up his technique and thus we find, as mentioned above, one
system extracting by the cold percolation process, and the
other extracting by the hot mechanical method. Modern
workers lean toward the latter procedure, insisting that
the alkaloids of Sol anaceae are not unstable.

Of the various methods described, the writer
decided to choose two to be used in the assays, in a compara¬
tive manner. The methods, also, were to consist of both the
cold percolation method and the hot, continuous method.

With a view of choosing the two most promising methods, the
above were considered in the order in which they appear.

(a) The B.P. 1914 method was used only on
Stramonium. It was discarded in favor of the B.P. 19 32
method for its advantages listed under (b).

(b) The B.P. 1932 Method. This method is out¬
lined on page 14 and was used in a comparative sense for
our plants and also as a check on various other methods.

The B.P. 1 9 32 method has several distinct
advantages and changes which tend toward greater accuracy
over the B.P. 1914 method. An examination of the two methods

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24

shows the following changes, with their advantages or
disadvantages •

i . Maceration - The B.P. 1914 directs "Into a

small stoppered glass percolator - introduce 10 grammes

of while the B.P. 1922 directs "Introduce 10 grammes

of powder into a flask". Both are then macerated for one
hour, but the B.P. 1932 powder must, of course, be trans¬
ferred to the percolator.

Maceration, by the two methods, it would seem
would be more complete in the latter, since it is much more
thorough to shake a flask than to shake a percolator, even
if it could be tightly corked. A second advantage of the
latter method is that no channelling can take place with
resulting incomplete percolation.

ii. Emulsification - Emulsif iciation in the
assay has been fairly well overcome in the B.P. 1932 method
by the addition of alcohol to the solvent. This, as pointed
out by Self (22), prevents emulsification, which occurred
commonly in the 1914 method.

iii. First acid extraction - At this point it
would appear that the extraction of alkaloids is more
complete since the ether- chloroform extract is run into the
acid. In the 1932 method, the acid is added to the extract
and shaken.

iv. Washing of acid solution - The B.P. 1932

directs that the acid solution containing the alkaloids be

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25

washed with chloroform, the chloroform run through acid
and then discarded, while this procedure is not carried
out in the B.P. 1914 method. This is a distinct advantage
since any coloring matter carried down from the original
volatile solvent extract is removed.

v. Washing of chloroform extract - The B.P.

1914 collects the chloroformic extract and directly dries
it, dissolves it in ether, dries and titrates it, while
the B.P. 1932 directs that the chloroform extract be
washed with water before being dried, alcohol added, and
titrated . This point is commented upon, as mentioned,
in the Pharmacy Journal (30), page 30.

In all determinations carried out in this
investigation, the final alkaloidal solution was washed
with water to eliminate any source of error in that direction.

vi. Titration - The B.P. 1914 directs the use of
0.05 N acid and 0.05 N alkali, while the B.P. 1932 directs
the use of 0.02 N acid and 0.02 N alkali. This is an
important step toward greater accuracy, and in some cases a
sharper end point.

vii. Indicator - The indicator cochineal, official
in 1914, is now replaced by methyl red, a much more accurate
and efficient indicator, as shown in the discussion by Self
(22), Evers (6), Mellon and Tigelaar (17) and others on

page 22.

In all determinations by the B.P. 1932 process, on
Belladonna and Stramonium, the percolator used was very
similar to that described in the U.S.P.X. method (Plate I,

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26

fig. 1). This percolator proved more convenient for small
quantities of drug.

In the extraction of Hyoscyamus by the B.P. 1932
process, the method is essentially that of Belladonna, with
modifications firstly for the large quantity of drug
necessary (since there is only a small amount of alkaloid
present) and secondly, the reported greater instability of
the alkaloids.

This is taken care of, in the case of the large
amount of drug, by using a larger percolator as shown in
Plate I, fig. 2, and which is of the standard type. Since
the quantity of drug is large, it is apparent that at the
acid extraction of the alkaloids, the volume will be large.
Before being extracted with chloroform, the mixed acid
solutions are made alkaline and evaporated in vacuo to
about 50 cc . at a temperature not exceeding 40°.

The alkaloids are shaken out with chloroform,
washed with water, the chloroform removed and the
alkaloids dried at 80° for two hours. This removes any
volatile bases present and so makes the final titration
more accurate. The alkaloids are titrated with 0.02 N
sulphuric acid and 0.02 N sodium hydroxide, using methyl
red as indicator.

In using the above method for Hyoscyamus, the
greatest objection to it is the length of time required
for completion. It is always advisable, where possible,
to complete an assay on the same day as started, since

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27

the alkaloids are of such a nature that standing over
night may completely change the final result. But with
the B.P. 1932 assay of Hyoscyamus, a complete assay is
impossible in one day.

The B.P. 1932 processes were used as outlined
in the text with the two following changes:

1. The liquid was percolated as directed, but
instead of adding the 0.5 N hydrochloric acid to the
percolate, the acid was placed in the separator prior to
percolation with the immiscible solvent. This was done
for two reasons:

(a) To avoid any possible leakage since the
acid layer would remain at the bottom, and

(b) To insure complete conversion of the
alkaloids to hydrochlorides.

2. All alkaloidal residues, on the addition of
the specified 0.02 N acid were warmed on a water bath to
insure complete solution. The flask was then cooled at
room temperature and the titration carried out.

The results obtained by the B.P. 1932 methods
proved quite consistent, and, as it is at present outlined,
the method may well be said to be one of the most efficient
of alkaloidal assays. Self (22), in his review of
alkaloidal assays of the B.P. 1932, stated that out of a
total of 46 alkaloidal assays in the B.P. 1932, 29 are
quite new and only three have remained practically as
they were. The changes, he says, are almost wholly of

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28

British origin, and it may be claimed that in the assay
section of the work the B.P. is stilj well in front. The
aims, of course, have been to make the assays simpler and
more acourate, but accuracy always remained the prime
object .

(c) The method outlined by Rosenthal er (see
"The Chemical Investigation of Plants" (1930)* was not used
in this investigation as it is a method for total extraction.

(d) The Method of Watkins and Palkin (27). This
method is essentially similar to that used by DeKay and
Jordan, which will be described later. The methods,
however, are different, in that Watkins and Palkin used a
specially devised apparatus while DeKay and Jordan used a
Soxhlet apparatus. Since the methods are alike, and

results may therefore be expected, and since the
DeKay and Jordan process provides certain advantages, the
Watkins and Palkin method was omitted in its favor.

(e) The Method of DeKay and Jordan (2). This
method was outlined by DeKay and Jordan after exhaustive
experiments, both on pure alkaloids and on the crude drug.

The extraction consists of mechanical extraction
by a Soxhlet apparatus, the drug having been macerated
over night.

The procedure follows:

Samples I and II (20 grammes each) in 60 powder
were placed in Soxhlet thimbles, dropped into place in the
extractors and moistened with a mixture of 7 cc. strong
solution of ammonia, 8 cc . of alcohol and 16 cc . of ether.

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29

It was well mixed with a stirring rod and macerated over
night. The following day, it was extracted with ether over
a water bath. The extraction required, as a rule, three
to four hours. The dark green extract, so obtained, was
evaporated over a water-bath, to about 1 2 cc . and then 8
cc . of 0.1 N sulphuric acid and 8 cc . water added. The
evaporation was continued until all the ether was removed.
At this point, the coloring principle was precipitated.

The liquid (acid solution of alkaloids) was filtered into
a separator and the precipitated chlorophyll residue re¬
dissolved in chloroform, 0.1 N sulphuric acid again added^
and again the chlorophyll was precipitated. The mixture
was again evaporated free of chloroform and the liquid
remaining again filtered into the separator through the
same -pafrer. The combined filtrate was made basic with
dilute ammonia (using litmus) and the alkaloids extracted
by shaking with successive portions of chloroform, until
completely extracted, as shown by a test with Mayer ?s
reagent .

The chloroform solution was evaporated to a low
volume and the residue dried at water bath temperature for
15 minutes. A brownish aromatic residue remained. This
residue was redissolved in chloroform, again evaporated and
again dried for 15 minutes. This process was repeated for
the third time .

At this point, the residue may be dissolved in
about 10 cc. of chloroform and then 15 cc . 0.02 N sulphuric

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acid added, the chloroform evaporated off and the titration
carried out, using methyl red as indicator, or the residue
may he treated for the removal of any volatile bases
present. The latter was the process used in this investiga¬
tion in most cases.

This process consisted of passing air through a
sulphuric acid wash bottle and a CaClg-NagCOg tower and
then through a CaClg U-tube, heated at 40°. This frees
the air of moisture and warms it. This warm dry air was
then slowly passed over the alkaloidal residue and
finally into the flask containing 0.1 N hydrochloric acid
(see Plate II, figs. 1 and £) .

The volatile bases thus carried over by the air
into the 0.1 N hydrochloric acid are there held. After
one hour, the hydrochloric acid solution is heated to
dryness and the residue weighed.

The a^katoidal residue is then titrated as
described above.

This method appeared to be a thorough procedure
for the extraction of alkaloids.

It provided a method for the hot extraction of
alkaloids and would, indirectly, indicate if the alkaloids
of the Solanaceae could be extracted by such a method
without loss.

The weighing of the drug into the thimble, the
maceration and extraction while in this receptacle without
the disadvantage of transferring from a flask to a
percolator, is obviously an advantage both from the views

of expediency and accuracy.

I . 1C '£

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31

The extract ion of* volatile bases (DeKay and Jordan)

A - HgSO^ drying bottle
B - Cadg-Na^CO^ tower
C - CaClg U-tube (at 40°)

D - A1 haloids'* residue (at 40°)
E - 0.10 N EC1
F - Trap containing HgO
G - To pump

Fig. 1

Fig . 2

Plate II

V'i;t .:-rov xO Xi: i;t03lJx9 ' IT

32

On obtaining the ether extract, the precipitation
of chlorophyll and other foreign extractives with acid
seemed an important feature, as the alkaloidal acid solution
should be in a comparatively pure state.

so the method of handling the alkaloidal
residue immediately prior to titration appeared to have the
advantage of bringing the alkaloids completely into solution
in the acid. This is pointed out by Self (22) who says
that if an alkaloid is difficult to dissolve in small
amounts of acid, the addition and subsequent evaporation
of a small volume of a solvent, such as chloroform, will
usually be found to facilitate solution.

Lastly, the means described of handling the
alkaloidal residue for the isolation and weighing of the
volatile bases seemed advisable in view of recent work,
particularly that of DeKay and Jordan, regarding these bases.

The whole procedure may be summarized as shown
on PI ate III .

After a few determinations with this process, it
was found advisable to make the following modifications:

(a) After the complete removal of the chloroform
by evaporation, the flask containing the alkaloidal acid
solution and the chlorophyll residue was cookd to room tempcmkre
whereby the precipitate was coagulated. Filtration was
then carried out by decantation. It was found that if the
solution was completely cooled after each evaporation,
very little chlorophyll was carried over.

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S3

Plan of DeKay and Jordan process
for a^kaloida! assays

I _

to acid layer

FiUer,

extract

— i —

make
wi th

al kal ine and
immiscibl e

'5UT7mr'TAYlR

ALKAL IKE - H ? 0

( alkal olds )

LAYER (non-al-

kaioida! matt®

L_

solvent and add
ings to solvent
_ _ _ _ I

wash-
1 ayer

Evaporate

ALKALOIDS and
VOLATILE BASES

Treat with warm
dry aiiy

Residue

Residue volatile

titrate

»

with air v^eigh

ALKALOIDS

VOLATILE BASES

Plate III

1x5 ' C u • . " to 3 ri

■

3 • .* \ 2 ' .{ 1:3 TOt

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. ■■

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* ub i a s

34

In all cases, after the original ether solution
was precipitated by the addition of water and 0.10 N
sulphuric acid, the precipitate of chlorophyll and other
non-alkaloidal extractive was quite bulky and on the first
filtration a considerable quantity of the tarry precipitate
was carried over on to the filter paper. This condition
was undesirable for three reasons, namely,

i. The probable loss of a portion of the
alkaloids in the precipitate, by being held back by the
gelatinous nature of the precipitate, and thus not being
thoroughly extracted.

ii. Much of the precipitate seeped through the

meshes of the filter paper and appeared in the acid solution

below, resulting, obviously, in a colored alkaloidal
residue, at the end.

iii. The precipitate in all cases closed the

meshes of the filter paper and made filtering almost im¬

possible, without repeated changes of papers.

(b) At the point of chloroformic extraction of
the ammoniacal solution of alkaloids, all chloroformic
extraction portions were passed through another separator
containing water. This held back any water-soluble foreign
substances undesirable in the alkaloidal residue, such as
traces of ammonium sulphate formed in neutralization. This
procedure agreed with a paper appearing in 1915 (30) in
which the writer suggested that a possible source of error

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35

in all alkaloidal assays is the carrying down of ammonium
salts, and recommended the washing of all final volatile
solvent solutions.

With these two main modifications, the process
was used throughout, as directed by DeKay and Jordan.

The process was used on all plants under
investigation in order to obtain a comparative alkaloidal
yield, after all volatile substances had been removed, and
to compare these yields with the B.P. process.

(f ) U.S.P.X. Method and Outline (29). This
method is essentially similar to the B.P. 1932 method with
the exception of the original technique, in that the drug
is placed directly into the percolator and there macerated
for one hour and then percolated. Since it was considered
that the U.S.P.X. and B.P. 1932 processes were so similar
that results should be comparable, the U.S.P.X. process was
omitted in favor of the B.P. 1932.

V. EXPERIMENTAL,

a. Test Garden

The test growing ground was a plot on the University
of Alberta campus, adjoining the gardens. It was normal
black soil, completely open to the sun, but well protected
from wind by an efficient protective growth of trees and
shrubs (see Plate IV, figs. 1 and 2). Pig. 1 shows the
north-west corner, and fig. 2, the east side, of the test
garden.

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The experiments'* garden

Fig. 1

Fig. 2

Plate IV

r

- 37 -

b. The Seed

The seeds used in this investigation were
obtained from various sources, in order to obtain offspring
of plants grown under different climatic conditions. The
seeds were obtained from the following sources:

1. The University of Minnesota, College of

Pharmacy.

3. Mr. I. Tice, Westlock, Alberta.

3. Seed obtained from the previous summer's
growth in this test garden.

In addition were used roots from the previous
year's growth.

c . The Growth

All plants were grown in the same test garden and
received the same amount of attention. The soil received
no fertilizer. The plants during their growth received no
special attention other than occasional watering and
weeding .

The moisture needs of the plants provided quite
a problem, as rainfall in central Alberta is not regular,
and during some periods in the l ifetime of Atropa Be"! ladonna ,
rainfall was completely absent. The plants, as mentioned,

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,

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38

were watered when necessary, and as far as was possible
without tests, were kept at optimum growing conditions.

1 . Atropa Bel i adonna .

i. First year growth. The seed, obtained
from University of Minnesota stock, was treated as suggested
with concentrated sulphuric acid, for 45 seconds, repeatedly
washed in a sieve with distilled water, and sown in the
greenhouse on April 28th.

While the seedlings were still very small, they
were transferred to the test garden, late in June . The
plants were late in developing, but they withstood the
severe frost (temperature 25°F.) which occurred on August
25th.

The plant produced several erect stems which did
not produce a particularly dense foliage. At maximum
growth, the plant was about three feet tall and produced
characteristic flowers.

The leaves were collected on August 26th while
at apparent maximum growth and when the plant was in flower.
They were cut by hand, and avoiding as much as possible too
much stem (the B.P. directs a maximum of 20$ stem) . Only
clean, entire leaves were collected.

The freshly cut leaves were placed in large paper
bags and without being pressed or unduly handled, transferred
to the drying room in the basement, quickly spread in a
thin, where possible single, layer on paper, on the floor.

The room was maintained at an almost constant temperature

of 50° .

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.

39

Upon being thoroughly dried, the leaves were
again gathered and ground in a Wiley power mill to a
uniform #60 powder, weighed, the whole well mixed, bottled
in amber, air-tight bottles and stored.

ii. Second year growth. All of the 1933 plants
allowed to remain in the field over the winter were winter¬
killed. Two plants which had been potted and placed in the
greenhouse the previous fall were set out in the field and
grew to a height of about two and a half feet. In spite
of the frost on August 25th and al so numerous September
frosts, these plants produced a few small seed-pods, two
of which ripened. The leaves were all so completely frozen
that they could not be collected.

2 . Hyoscyamus niger .

i. First year growth. The seed for the 1934
grov/th of this plant was obtained from Mr. L. Tice of
Westlock, Alberta, from plants grown by him in 1933.

The seeds were treated with concentrated sulphuric
acid for one minute and 45 seconds before sowing.

Sowing was carried out on May 8th. The seedlings
appeared about May 24th. The growth of the plant was fairly
rapid.

The plant had a characteristic fetid odor and a
heavy growth of the characteristically indented leaves was
produced. The plant as a whole was low in stature with no
upright stem.

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40

The size of the leaves varied considerably. They
were of a pale green color and while growing, had a soft
and unpleasantly clammy surface. This stickiness was due
to the soft hairs found near the veins, particularly on
the under surface, which possess glandular heads secreting
a resinous substance.

Two collections of leaves of the first year
plants were made, the first on August 1st and the second
on August 10th.

They were transferred to large paper bags and
thence to the drying room where they were spread evenly
and dried. They were then gathered, ground to a #60 powder,
well mixed, bottled in well-closed containers and stored.

ii. Second year growth. The plants of the
second year were grown from the roots of the first year
growth, allowed to remain in the test garden over the
winter, so that the conditions for growth for both first
and second year plants may be said to be the same.

The roots of the plants sown in 1933 (which was
L. Tice stock) left in place over the winter as mentioned
above appeared to winter well.

The young shoots appeared above the ground about
April 24th. The roots, soon after, were thinned out by
transplanting, this work being carried out between April
27th and May 3rd.

The growth, as in the first year, was fairly
rapid, but in this case the plant was considerably bulkier,

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41

with a greater number of leaves and with a strong upright
stem. At maximum growth, the characteristic flowers
appeared in large numbers.

The plant, at the period of flowering, attained
a growth of slightly more than three feet as may be seen
in Plate V, fig. 3. The leaves had the characteristic
indentations and were of a dull green color (Plate V,
figs. 1, 2, 3).

On July 20th, the leaves were gathered, keeping
in mind the limitations as to stems, directed by the B.P.
The leaves were cut closely, transferred to large paper
bags and spread in the drying room. When thoroughly dry,
they were broken and passed through a Wiley mill to produce
a #60 powder, thoroughly mixed, placed in well-closed
containers and stored.

3 * Datura stramonium.

The seed for this plant was obtained from the
University of Minnesota stock. It was not treated with
concentrated sulphuric acid, as was Atropa Bel 1 aflonna, but
was sown directly into the test garden on May 18th.

The growth of the plant was fairly rapid. The
stems were round, very smooth and at maximum growth, woody.

The plant appeared to be more hardy than the
others, and the growth seemed to bear out the statement of
Koch (14) who stated that the plant required very little

attention.

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.

Hyoscyamus niger (second year)
Flowering plants

42

Plate

43

The pT exits, however, were cultivated as regularly
as possible and watered occasionally. The growth of weeds
was kept at a minimum. The plant produced quickly an
abundant growth of light green foliage, with its deeply
indented leaves and foul-smelling odor.

It grew to a height of about three feet, when the
flowers appeared with their funnel shaped corollas. In
rapid succession, the flowers gave way to the large 4-cel led
prickly walnut-shaped fruits.

The leaves were collected on August 16th, trans¬
ferred to the drying room, dried, collected and ground to
a #60 powder. The powder was well mixed, transferred to
air-tight containers and stored.

Shortly after the leaves had been gathered, the
life of the whole plant ended and the fruits began to open.
The whole plant was then cut down, gathered, dried and the
seeds threshed from the open capsules and a large yield of
seed was obtained.

4 . Datura Me tel.

The seed was obtained from stock of the University
of Minnesota, and sown on May 18th.

The growth of the plant was rapid and produced an
ahundance of leaves of a yellowish green co*»or, which were
irregular in shape. The odor of the plant, like Datura
Stramonium, was unpleasant and characteristic. On the
epidermis of the leaves occurred scattered simple as well

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44

as small glandular hairs. At maximum growth, the plant
produced white flowers.

No cultivation, other than occasional weeding
and watering, was carried out on the plants.

The leaves were collected on August 1 6th in much
the same manner as Datura Stramonium and the other
solanaceous plants.

Here again, as little stem as possible was
gathered and only the healthy clean leaves were used. The
leaves were dried in the drying room at the constant
temperature. When dry, they were gathered, ground to a #60
powder, well mixed, bottled in air-tight containers and
stored .

d. Results of Determinations

1 . Atropa Belladonna.

As will be noticed from the tabulated results,
leaves of various years growth were used in the determination

Since a considerable amount of 1931 and 1932 leaf
was on hand, it was used for the first several determinations
until the technique in alkaloidal assays was perfected. The
final results were taken from determinations carried out on
leaf of 1933 and 1934 crops.

The first determination of 1933 leaf was that of
moisture content. Then a determination of ash content was
carried out, and finally alkaloidal assays by the Delia y
and Jordan method and the B.P. 1932 method.

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■ . . •

45

1. Atropa Belladonna (first year)

TABT.E II

Moisture and ash

content

Crop

Moisture

%

Ash Remarks

%

1933

1933

1934

1934

7.795

8.298

6.954

7.0923

12.7155

11.8155

TASTE HI

Alkaloidal content

Date

Crop Method

Alkaloids

%

Volatile

bases

E

May 12/34

1933

B.P

tt

tt

tt

May 15/34

t»

tt

tt

tt

tt

Jan. 4/35

tt

tt

tt

tt

tt

tt

1934

tt

tt

tt

n

Jan. 10/ 35

1933

»

n

tt

tt

n

1934

tt

tt

tt

tt

Jan .16/35

1933

DeK.&

tt

tt

tt

tt

1934

tt

tt

tt

tt

Jan. 18/35

tt

tt

tt

tt

«

0.2448
0.2873
0.4376
0.4446
0.3688
0.3617
0 .3633*

0.3798
0.3454+

0.3400+

0.3410+

0.3394+

Jord. 0.3293**

0.3275**

0.3314++ 0.0045

0.2560++o 0.0035

0.3294**

0 .3306 **

* At ether-alcohol addition to leaf, became almost
solid. 1934 leaf evidently drier than 1933.

+ Residues not dried. Titrated by DeKay and Jordan
method. Evidently lowers results.

** Volatile bases not removed and residue not divided

++ Volatile bases removed but residues not divided.

0 Partly lost.

** volatile bases not removed and residue not divided

Snedneo

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•: ' h' r

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46

2. Hyoscyamus niger (first year).

The stored powder representing Hyoscyamus niger
(first year) was well mixed by turning it out on clean
paper and alternately lifting the corners. It was then
mixed with spatulas and replaced in the containers, from
which samples were taken for the various determinations.

2. Hyoscyamus niger (first year)

TABLE IV

Moisture and ash content

Crop

Moisture

Ash

Remarks

%

1o

1934

10.768

13.515

TABLE V

Alkaloidal content

Date

Crop

Method

Alkaloids

%

Volatile

bases

<

Feb .22/35

1934

B.P.

0.03413

t»

tt

tt

0.03430

tt

ft

DeK.& Jord.

0.0664 *

0.0056

ft

tt

tt

0.06312+

ft

tt

tt

0 .06443*

0.0084

ft

tt

tt

0.06434+

Feb. 26/35

tt

B.P.

0.035015

ft

tt

tt

0.03378

ft

tt

DeK.& Jord.

0.06263

0.0072

ft

tt

tt

0.06189

0.0136

+ Volatile

bases

not removed

•

* Volatile

bases

removed .

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47

Hyoscyamus niger (second year)

TABIE VI

Moisture and ash content

Crop

Moisture

Ash

Remarks

%

%

1934

11.236

14.587

n

10.940

14.590

TABLE VII

Alkaloidal content

Date

Crop

Method

Alkaloids

%

Volatile

bases

%

Feb. 1/35

1934

B.P.

0.02145*

Feb. 5/35

tt

tt

0.04741

tt

It

tt

0.04727

Feb. 7/35

ft

«

0.04110

tt

ft

w

0.04147

ft

tt

DeK.& Lord.

0. 06034*

0.0047

tt

It

tt

0 .06080*

0.0056

Feb. 7/35

tt

tt

0.07899**

ft

tt

«

0.08012**

Feb. 14/35

ft

B.P.

0.0438

tt

tt

tt

0.0433 *

ft

tt

DeK.& Jord.

0.0763

0.0040

tt

ft

tt

0.06052*

0.0068

+

Check lost.

*

Volatile bases

removed .

**

Volatile bases

not removed.

a>.-T • • e '

'

. r

0~ . r

^ r

.:. ‘ • r J

i'I: rc ■ : r. '" r..": r;.

\.

r v , “

,

orr;i . c

,

*

.

; r . : . :j

,

,

.

> :g f

tf

. 'r’

ft

■

ft

,

'r? Ht

r .

.

ff

;

.

- V r .

t*

.. - r.

. j: ; ' r jforsrfC

. : 5 : r 1 . r C

■*

' ■ r '

3. Datura Stramonium

TABLE VIII

Moisture and ash content

Crop

Mois ture

%

Ash

%

Remarks

1932

4.755

13.8875

Water insoluble

1934 .

5.4800

13.852

ash 49 .3534%

TABTE IX

Alkaloidal content

Date

Crop

Method

Alkaloids

Volatile

%

bases

%

Feb. 15/32

Penick

B .P.1914

0.3013

tt

tt

tt

0.2526

Mer. 1/32

tt

tt

0.28562

ft

tt

tt

0.27612

tt

Local * 31

tt

0 .18346*

tt

tt

tt

0.1822 *

Nov. 23/ 34

1934

B. P.1932

0.2545

ft

w

tt

0.2519

Nov .28/ 34

tt

DeK.& Jord.

0.20115

0.0170

»»

«

«

0.19805

0.0100

Dec. 14/ 34

tt

B.P.1932

0.20356

tt

tt

tt

0.1884 +

Dec. 18/ 34

tt

tt

tt

DeK.& Jord.

tt

0.2982

0. - **

0.0125

Dec . 26/ 34

tt

B.P.1932

0.170 ***

tt

w

tt

0.154 ***

tt

«

0.22512

tt

tt

«

0.22512

tt

tt

DeK.& Jord.

0.2529

0.0120

tt

tt

n

0.2532

0.044

* First Alberta crop.

+ Partly lost due to faulty separator.

m* Sample B lost completely, A divided and part with
volatile bases showed higher alkaloid(.302 q pggc

).2944

*** Results low carried down considerable coloring matter.
Titration difficult.

•j j. 0 A 3 8 birti 91 J 'J'J -• i oM

r

. .

3riifSal; croiO

O'^.S - [

r

r

■c3. '

■ .

r.

' r.

v: r. :

v. Ta;; . 0

r r .

gc^Oa.

Ad r ,

- .

QVJLO
gcI.C
,• ro ■ A . o
r . . .

• 1 G H . ' ■

... ,

botite.l qciO etfad

rv r.

n

4f

ft

r t

. i ' C , '

'

r. .

,

ft

■AG A r/:j .£

. '

t >q

n

ft

tt

£ * £

«

-■£*> r

tt

n

♦t

tt

tt

tt

H

ff

ft

ft

ft

r.

5\t .08

tt

.

.

It

• V r. :

: , er.of-u

a : > ,...■ ; . v •<

11

ft

tt

. c ; o id .6 $ *1 e cf r 2* 3 a ixg

,

.

- - . , /

***

f ; g?Id rw< a ..

. :j r:jai 1‘ii.o ru i Ac^icT IT

49

4. Datura Metel

TABLE X

Moisture and ash content

Crop

Moisture

Ash

Remarks

*

. %

1934

7.104

14.99

tt

6.978

14.9112

TABLE XI

Alkaloidal content

Date

Crop

Method

Alkaloids

fo

Volatile

bases

*

Feb. 7/35

1934

B. P.1932

0.2050

tt

tt

tt

0.2144

Feb. 8/35

tt

tt

0.2138

Feb. 14/35

tt

DeK.& Jord.

0 .1947'*

0.0088

tt

tt

tt

0.2389+

«

tt

tt

0.2234*

0.0076

tt

tt

«

0.2263+

Feb. 15/35

tt

w

0.2270**

tt

tt

tt

0.2291**

Volatile bases removed.

+

Volatile bases not removed.

Residue not divided and volatile bases not removed

r -

.

..

a8i:.

* r
r>. , ■ r

rwJto'

r.

r

r o r ..

■ ■- v

8©SBCf

..

. .

i c r : . r.

.Don ere.

,

r .

*

,

**0VS3, o

r . . o

.

ft

n

ft

.

- . . .

it

: r • . r .

: ■ . : " ' ’ rCv

'

o, a

50

VI. DISCUSSION

a. The Plant

In all cases, the plants under observation grew
and matured with a growth similar to that described for
the official B.P. plants found in southern Europe.

It was found that:

1. All plants were free of insect life and

mi Idew.

2. The roots of Atropa Belladonna, at least,
cannot resist the winter climate, in the field. They
should be dug up and kept dormant in a warmer storage house
and replanted in the spring for the second year*s growth.

3. The roots of Hyoscyamus niger , left in the
field, wintered well. Seeds planted for the first year on
May 8th produced, from the wintered roots, shoots on the
following April 24th. The plants produced by these wintered
roots were hardy and had a healthy, rapid growth.

4. Datura Me tel is the least suitable of the
plants under observation, for Alberta. Only one plant
produced seed pods, though it is an annual. All plants
grown in the greenhouse, produced seed pods.

5. Atropa Belladonna must be started in the
greenhouse, as the roots, as mentioned above, are killed in

winter .

IG

¥

J :

r

r

. .

itBiif bnuclt sbw SI
slit ' ■ fq II

. wsbf tm

c

. •• r , , « -}:-uTo ’ .-• .i 1 ■■ ' : •: •

f

. r

f

. - ; * r r 5 . i i v < . r ■ n

... ,r. ; 3 v : x .. 3 AS aeri < l» b Duhotq rfcfB x^U

r

. i . ( t r . : if I

'

3 ■ t : r ! :>y.rc V: :■ . 33 -:.^d r„ -■ .3 . - i - v-: ••:••; 3 ;v r.o

'

. BiK 38 . . ? nl

-

,

51

6. The roots of Hyoscyamus and Stramonium are

hardy.

b. The Results

1 . Moisture contents and ash values.

Ash contents were fairly consistent and all
remained within the B.P. limits as shown. Various other
values are given for ash values as follows:

Atropa Bell adonna - Youngken* "Pharmacognoscy" ,
maximum 20%.

Datura Stramonium - Youngken "Pharmacognoscy" ,
maximum 20%.

In all cases the results obtained will be tabulated
against the standards required by the B.P. and various other
results, in a comparative manner.

TABLE XII

Moisture and ash values of solanaceous plants

pi ant

Crop

Moisture

Ash

1o

B.P. ash
require¬
ments

Atropa

Bell adonna

1935

8 .047

12.7155

Maximum 15%

ft

1934

7.0232

11.8155

rt

Hyoscyamus niger (1st year)

1934

10.7680

13.515

n

rt

n (2nd year)

1934

11.088

14.589

tt

Datura

Stramonium

1932

4.755

13.8875

Maximum 20%

ti

tt

1934

5.480

13.852

tt

Datura

Me tel

1934

7.041

14.9006

*

Published by P. Blakistonfs Son & Co., Philadelphia, (1921)

*

. Y&'icjrf

r

# ,

-

'7 7 0 0 >7'a;i

. r

rr7 L 73

f

i .» -ii 9*1 6W

1777 7 axici'ic V

. .

a 8

... it .s 1 i 1 X 1 »

,

r

;j ■. P‘T : 7 -■-=7 7 8 7,7r.-::V

•‘,oe<: . mo—O:

• ■ < •■>• ... *?

- 7'7'7' :

, r r . . • ;j-

. •. , . , '• . ...7;. ■ ■ ~ : :J . _;00;v...

,705 liixjmixaxa

a, ■;•- 7/n .af T f .7 :; b Si! i o 0 C C- 8 J Sdt 7 37 '0 r r:l ill

, *

: : / .: ' ■ . : 7": : :T i { 8j r L* ■' ■ 8 1

I IX

.T

r

. .

la . &

luloio l

qo‘iO

1 0 7 r ,

-81X77 t 7

Su

. .7 0 . ., . __

- r ■

■ " . _

OO'O.S

,77 r

7,7777 '.77 r r •’ .'■ 0 7 77i -77.0.

7 G r 7- * 'r

OOSOaO

o-.-o: r

5?

o ra . 7 r

o 60 v . o r

■

( is.; sx

r i 777 Zil 7 7773 10807 1

.77 7, ?■ r

MOT

(?B»X

fin

j IT «

r Ots lOil-iXal

0 777a 7 r

'.70.

iilL'i OCO’lO 10 . 71 70 30

rt ■

007,01

,

7^00 r

w t»

, r

r - , V-

r-7>:0“ 7173 7,7

, ; rs-Qf j ... .7

f. Cioiri

, - 01 _

no8 a r

77 0 7 7,7 lO:

,

52

Due to the slight variations in the moisture
content of the drugs, and particularly to the urgent need
in the Department of Pharmacy for an efficient drying oven,
it is suggested that an oven with a maximum tempera ture
range of 100° and capable of accommodating considerable
fresh drug, be constructed.

The efficient drying of a drug is a very

important factor in atkaloidal determinations as it has

been suggested (Bentley "Text Book of Pharmaceutics"*

page 260) that the alkaloids exist in the plant in the

form of alkaloidal tanno-glucosides , which undergo partial-

decomposition when, on the death of the cell, the

permeability of the plasma membrane changes and the acid

cell sap enters and forms alkaloidal salts. This reaction

is not complete, so that it is necessary to completely

separate the alkaloids from the primary colloids. The

decomposition in most cases takes place by means of the

enzymes present, and these act best at a temperature of
o

about 50 C.

* Published by Bailliere, Tindall and Cox, Tondon.
(1933) .

H-r 3.'.- ■ ' C £ i \\ 3 ’ J i r< '1 Oj 0 .C/C

r

t v-u Ssr-jic i. tie ns 101 ^oBicxiafn ‘io Jnaifi^nsqaCI ©rftf ni

itruiTtxxBin s - j iv. rtevo its 3&M3 bs^sesgi/s si ;ti
td" ter o v j 1 ■■: c r : ' ' 0 )f >| .

. •• ; ^T3:;oo ©d . r*r- 3 ffseTl

/lev a i auif) a ic viiv-io :;n to - ri9 e-.I?

r

f

c

r - ' r r
< T r

LOB S 0 • '

• . : r..; : r r x . •. r r c

f

. Ol B2LC B

. : i o 8 xtB 3ivi 4 cf &ps.C q 3 : • ; • t? 0 3 s 01 : til nci 3 i: e o qmo 0 a b

v., red' a is serf uos ex.: r a tixtea©iq semv^ne

o

. 0 Go 3 nod s

• jv curve T

. xoU 'a\a r f a: x. i.i f s-ioi r

iaS

r

(seex)

*

53

2 . Alkaloidal contents.

TABLE XIII

Average alkaloidal contents of Solanaceae (B.P. method)

Plant

Crop

Method

Alkaloid

%

B.P.

require¬

ment

Other

references

Atropa

Belladonna 1933

B.P.

0.3797

0.30

0 .40$*

" 1934

0.3059

0.30

0 .40$**

Hyoscyamus niger 1934
(1st yr

tt

.)

0.0343

0.05

0.063$*

tt

" 1934

( 2nd yr

tt

.)

0.04406

0.05

0.093$**

Datura

Stramonium 1934

ft

0.2321

0.25

0.22**

Datura

Me tel 1934

tf

0.2111

—

0.40$*

0.50$**

* Henry "Plant Alkaloids", Published by J. and A. Churchill
London. (1924).

** Greenish "Materia Medica", Published by J. and A. Churchill,
London. (1920).

TABLE XIV

Average alkaloidal contents of Solanaceae ( DeKay and

Jordan method)

Plant

Crop

Method

Alkaloid

%

B.P.

require¬

ment

Other

refer¬

ences

Atropa Belladonna

1933

DeK .& Jord.

0.3284

0.30

tt tt

1934

tt

0.3305

0.30

Hyoscyamus niger

1934

tt

0.06342

0.050

(1st yr.)

Hyoscyamus niger

1934

" (bases

0.06449

0.050

0.0868$

(2nd yr.)

removed)

DeK.& Jord

Hyoscyamus niger

1934

" (bases

0.07956

0.050

0.1313 $

(2nd yr.)

not removed)

ibid.

Datura Stramonium

1934

DeK.& Jord.

0.2407

0.25

" Me tel

1934

" (bases

0.2208

removed)

tt «

1934

" (bases

0.2268

not removed)

,

.

r

'

'I:?:.- J O

r

r' ' ' .

r

; : r

r ; r: ' r ■■; v. oca;

f •;

r

?r bilS )

O V., r . i. ■ : OTJO J'iiJJ

: c- r re?5 : om/GsG

t T flfiuT

ic

! * ^ tit 9H

. no fine t

a fills

d u1

i

.

[

fa o d f e m f ( .3 b id.

,

*

an ft c r fad aqcTCfA

j.o G. •'

0 la r

$ ? f f

:GoGo(G 0

ft

*163 i n s JLrnrs'G o e o^H

■

8 9KBC } f?

r

•; 1 ' .

! . tv GoG )

)

(1)0 -/MOST >tOiI

G ’ r

too i ■ ■ b Jiiio b o a ov, :
(.to GfiG j

' -

Bin*

308BO;

( b evened

oo G r

roi'O t

*

G 08 0 0 j "

{ COVGn.GO' 0 Oil

0 C' r

*t tt

54

2 . Aikaloidai contents.

A comparison of the results obtained on the
plants by the two methods with the official requirements
and results obtained by some other workers shows that by the
B.P. 1932 method,

1. Atropa Bel i adonna is the only plant which
produced alkaloid in excess of B.P. requirements, and very
little lower than percentages tabulated from various reports
in the text "Plant Alkaloids" by Henry (9) and "Materia
Medica" by Greenish.

2. Hyoscyamus niger (both years) falls con¬
siderably below the B.P. requirement.

3. The second year Hyoscyamus growth is much
richer in alkaloid than that of the first year.

4. Datura Stramonium falls very little below the
B.P. standard, but is richer in alkaloid than the
commercis1 samples which Greenish reports as containing an
average of 0*22% alkaloids.

5. That Datura Me tel (which did not produce seed)
falls considerably below other standards as shown in the
tabi e .

The same plants when assayed by the DeKay and
Jordan method show that:

1. Belladonna and Hyoscyamus both show an
alkaloida1 content considerably in excess of the B.P.
requirements .

2, The second year’s growth of Hyoscyamus is
considerably richer in alkaloids than that of the first year.

. . - • :■ ■ ; " r A.i C Aa

■ i3 r .& r DA ' I A- ' *1 : 1 "i mov A

. ... .. r , .. ; ;; , , : .. . ,A ■. : ovA t : s^'b A

'

t I J* ' A . A r . ' . S

A: ; ' - ■' .laaaaaLQa; * r

1 t r •%* ■

r-®r

r

, ••. I • a\ ■ , o ’• .so 1 .c :A;r

- e r " • ' re .■ . T^;v in C'i'inzxozc'Clj

■i : ; ■ ": : ;• •'■•: , A . A: r ’A." r ’ : r:. f~ T- J 8

#

r.d

, ... . - . a - - • i ■ : •, , : At -A. j -a . aaaa . A-S

i

; . -• .- V r ' ' ' '

. . A 3$

... . ..'.A .. . ,.7V. /A I. ATT r' OniBB - A'

;;} .. 1 Doiijan n&biol

A . : A: ,A(' ' ... A ; ,aaa T- ... " rA

: v.-. : ; a, -AT A A a t ; A AAA. -- r-,.: A,

.

*

t , ., A' a a. A -A..I - A..r-.: A; xti 'AAiA v r ci A Ab i snoo

55

3. Stramonium falls just below the B.P. re¬
quirements, and

4. Datura Me tel , considerably below commercial

samples.

Analysing then, the various tables, the following
results are seen for the various plants.

TABTE XV

Alkaloidal contents of solanaceous plants when
assayed by B.P. 1932 and DeKay and Jordan methods

Crop

Atropa

Bella¬

donna

Hyos-

cyamus

niger

first

year

Hyos-

cyamus

niger

second

year

Datura

Stra¬

monium

Datura

Metel

Growth

1934

Develop¬
ed late

- Abun¬
dant
growth

Fairly

rapid

and

heavy

Hardy

and

rapid

Flower¬

ing

slow

Ash %

B.P. maximum

1934

11.815

15.00

13.515

15.00

14.589

15.00

13.852

20.00

14.9006

Moisture %

1934

7.0232

10.768

11.088

5.48

7.041

Alkaloids

B.P. method

tt

DeKay & Jordan
method

B.P. minimum

1933

1934

1933

1934

0.3797

0.3059

0.3284

0.3305

0.30

0.0343

0.06342

0.050

0.04406

\ 0.0645
0.050

* 0.2321

0.2407

0.25

0.2111

0.2208

From this composite table will be seen that:
1. In all cases Atropa Belladonna contained

alkaloid in excess of B.P. specifications.

r ' r

‘1 .

■: : . :.r “ v . ..

(■ C

r r

T^d ba^B83B

r •'»

'

6 C ■£’■ ■ »

*cQ

r

■r: r

: - r

r

1' • ' r - ' rA

5c . ' a it # ,

•o^ici .$ ^B-lad
'to 'St ~-oi
pur -yd .

r ri

. rrj., ' rr ,'U . r

3890X0

r . rc

56

2. Stramonium and Datura Me tel are below the

usual , as shown by both methods.

3. Hyoscyamus is below the requirements by the
B.P. process and above by the DeKay and Jordan process.

4. Ash value in all plants is within B.P.

1 imits .

3 . Volatile bases .

In order to determine what effect, if any, the

separation of volatile bases had on the remaining
alkaloidal residue, several determinations were carried out
with the following technique:

Two check samples A and B were weighed out as

usual and the regular DeKay and Jordan extractions cerried
out to the point of the last chloroformic alkaloidal
solution. At this point, instead of each solution being
evaporated down with chloroform and acid, three times, the
solution was transferred to a graduated 50 cc. cylinder and
made up to 50 cc . with chloroform. Each sample was then
accurately divided in half, each portion being transferred
to a separate flask. Thus the samples became

A^ (25 cc}

Ag (25 cc}

B^ (25 cc.)

Bg ( 25 cc}

each flask representing one-half of the original sample.
The A^ and B^ samples were each freed of volatile bases,

- V: -

r r ;;

. r - .. : i: aunj.^v osc : . ♦ %

- .

, , £ t r t r *

.80 mi r

, v ' " " rj :C'J.rcj" *

e j xl w.m w. j. , *i m c:r *i eb-ic ."1

i. sm .. semo o fm rov 'lo itcid'a‘i3.Q3S

. r

: -7 i: .’osi - : • ; r fo •? !:■ c-i-
' : : ~ .7 c

”• r *•' 0 0 -7 7 r .737 r

r r r T • r

.; t . loTOJTdo rffrlw mob beiBiQqevQ

■:■■■ -r rv .00 Co be

/ 3 . . : - r ■■ •

•/ ot .uofi -to i on -01^ sow nolto

'

IOlO*IC

. s Vi - '

: ■ - t fa

■ r . : ■ ■ . 37 7 .. T . C: : r7 " ~ r: w or: C t

*- C ; r

i.

* ■ 0 ■ ' '

>a

sa

'.no as)

(.00 <5S )

, l . ' r . r

r ‘ . I-

. i r ; ; • r- . ;-i'i ;. ” e r ■ .. ' i

57

while Ag and Bg were titrated directly. Diagrammatical"? y
this diviaiQn and titration may be shown as

Sample A
I

total chloroformic extract (50 cc.)

i - - ^ - 1

A-j_ (25 cc.) Ag (25 cc.)

i

warm dry air

| ^ I titrate

titrate weigh

I *

Alkaloids Volatile bases Alkaloids

The DeKay and Jordan process appears to remove,

by use of the warm air apparatus, all the volatile bases

present. This can be seen by consulting Table VII, page

47, where on February 7/35, two determinations were carried

out and directly titrated without extracting the volatile

bases. The results obtained were:

0.07899%

0.08012%

Average 0.07956%

This result compares favorably with the other
results in which the residues were originally treated with
warm air, and gave an

Average 0.06449%

In the determinations in which the residues A and
B were equally divided into A-[, Ag and B^ , B^ as outlined
on page 56, it was found that the portions titrated without
volatile bases removed were consistently higher as evidence

t

(.00 05}

JO

3

r>\ «r>,

.00 ' ) r

-ii.3 fib m i.s io

g ^

r v

o-t eiBeqqB

r .

• , a u*

*xcT* on - ->G. snT

•-*. • ;• re o 1 3 ■ tin or a;i ;t ’i c oa u yd

r

■

r i x rov S'. 1 ' •• i‘i o.r; j ucn'J

n

\o midst no ©lo.iw
■: •; io v; rd'CS,Tl£> DiT.6 &vq

isirw b 9ii

".■ T r C> ■'

>s&d

f

V.- iS- .

j

3 O ‘

31

r

b il^fes

Juc;

£*0.11'

a sorb is si . erl^ iiox/lw nl
ir.;ec. 03 .L r

f& ' )i io<

VK 3 ri ■ 3 b .1.

J iz. 3 v 3 f 5 30 t -> i 3 mfl£W

. , ( . i non

a n c jt ^ exi x one o s b a ri $ n I

r

.■ tjjlx bnuc-l asw cM fad amsq no
n aw ik v . . K : n eo e ri r ; rc. r

Table V (page 46) in which the four readings of two
determinations are recorded.

Sample 0.06312%

z'.

A, treated with air
0.06476% 1

Ag not treated with air

B1 0.06434%
B2 0.06443%

B treated with air
0.06438% 1

B2 not treated with air

Here the average alkaloid when bases are removed is 0.06373%
and the average alkaloid when bases are not removed is 0.06543%.

Since this investigation was interested

essentially in total alkaloid determinations, the
volatile bases and other residues brought down with the
alkaloids were not identified. But in view of the various
reports on substances brought down, it would prove an
interesting investigation to enquire into the exact
nature of all compounds extracted during a normal assay
by the B.P. or mechanical methods, of a solanaceous
plant. It may disclose many compounds brought down,
which at present may be the cause of such various and
differing results obtained, as pointed out by Sievers (24).

As mentioned, DeKay and Jordan (2) outlined

their process with the object of isolating and weighing
the volatile residue, which they definitely determined as
di- and tri-methyl amines and pyridine. Goris and
Larsonneau (8) isolated from Atropa Belladonna many
volatile bases, an aliphatic amine, pyridine, N-methyl
pyrolline, N-methyl pyrollidine, etc., all present in
small quantities. It is possible, moreover, that many

more compounds exist in the alkaloidal residue, besides

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the above mentioned.

o. The Methods

Finally, the methods used may now be critically
examined, with a view of determining their values or
disadvantages .

1 . The B.P. 1952 method.

There is no doubt that the B.P. 1932 assay
methods for the solanaceous plants are tremendous improve¬
ments over the B.P. 1914 methods. The advantages of the
former over the latter have been fully dealt with on
pp. 23-28, the only factor in the procedure which may lead
to error is the technique of titration.

All alkaloidal residues should be warmed slightly
on a water-bath, after having had the addition of 0.02 N
sulphuric acid.

All alkaloidal assays by the B.P. process carry
down small amounts of colored residues, and if this be
warmed while in contact with the acid, a greater degree of
solution of alkaloids is affected. It should be then
cooled to room temperature and titrated.

2. The DeKay and Jordan method (2).

This method in all cases save Atropa Belladonna
1933 crop (see Table XV, p. 55) gives a much higher
alkaloidal content than does the B.P. process. This may be
due to a number of factors.

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i. Maceration. The extraction of the alkaloids
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complete, since maceration is carried on over night. Also
there is no chance of loss of drug, since there is no
transfer from flask to percolator as in the B.P. method.

ii. Extraction. The extraction of freed
alkaloids should, by this process as well as in the B.P.
process, be complete, since the extractive is tested for
absence of alkaloids in both cases.

iii. Titration. This process appears to have
the better technique as applied to the titration of
afkaloidal residues. There should be complete solution

of alkaloid, and therefore complete titration of alkaloids
present .

Though, as reported, various workers have had
difficulty in obtaining consistent results by various
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VII. SUMMARY AND CONCLUSIONS

Observations and determinations were carried out
on the following Alberta grown plants:

Atropa Belladonna

Hyoscyamus niger (first year)

Hyoscyamus niger (second year)

Datura Stramonium

Datura Me tel

with a view of determining with what success these plants
could be grown in central Alberta, from the viewpoint of
both growth and alkaloidal content.

For the determination of the alkaloidal content
of the various plants, two methods were used: (a) The B.P.
1932 method being an example of a cold percolation process;
(b) The DeKay and Jordan method, being an example of a hot
continuous extraction.

From the investigation the following conclusions
were drawn.

1. The above plants can be successfully grown
in central Alberta.

2. The roots of Atropa Belladonna cannot resist
the winter climate.

3. The roots of Hyoscyamus niger can resist
winter climate.

4.

The moisture contents were fairly uniform.

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5. The ash values were within the B.P. limits.

6. Atropa Bell adonna produced alkaloid in
excess of B.P. requirements.

7. Hyoscyamus niger (both years) falls below
the alkaloidal requirements of the B.P. except by the
DeKay and Jordan method.

8. Second year’s growth of Hyoscyamus niger
is richer in alkaloid than that of the first year.

9. Datura Stramonium falls just below the
alkaloidal requirements of the B.P.

10. Datura Me tel falls below the results given
for commercial samples, and the growth is not particularly
successful in Alberta.

11. By both methods used, and from results
obtained, the solanaceous alkaloids are more stable than
they are variously reported to be.

12. To avoid inaccurate results, plants contain¬
ing volatile bases should be treated for their removal.

13. Using both methods on the same plants, the
DeKay and Jordan method shows a much higher result, due
probably, to more complete extraction.

VIII. ACKNOWLEDGMENTS

The writer wishes to acknowledge his indebtedness
to to. A. W. Matthews, Assistant Professor of Pharmacy at
the University of Alberta, for his valuable suggestions,
criticisms and assistance given during the period of the
investigation .

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IX. BIBLIOGRAPHY

BORNEMAN, I. A. Successful cultivation of medicinal
• plants in Pennsylvania. Am. J. Pharm. pp . 546-
554. Dec. 1912.

DEKAY and JORDAN. The assay of Hyoscyamus. J. A.

Ph. A. p. 316. Apr. 1934.

DOHME , A.R.L. A.S.D. XXI from Proc . A. Ph. A. 1894.
p. 231. 1094.

EVANS and DAVY. The assay of Hyoscyamus. J. A. Ph.

A. p* 388. May 1934.

_ and GOODRICH. Washington Belladonna and methods

of assay. A. Ph. A. Sci . Sect. p. 824. Sept.
1933.

EVERS, NORMAN. The titration of alkaloids. J. A.

Ph. A. p. 676, and J. Soc . Chem. Ind. XI: 233.

1921.

GORIS and DETUARD. Bull. Sci. Pharmacol. XXIV: 74.

__ and IARSONNEAU. J. Pharm. Ghim. 23:175.

(Br. Ph. Ass’n) . 1921.

HENRY, T.A. The Plant Alkaloids, p.63 (text). 1924.

KARR, E.H. The effect of cultivation on the alkaloidal
content of Atropa Belladonna. Chem. and Drug
81:432. 1912.

_ . Cultivation of Belladonna at B. W. Materia

Medica farm. J. A. Ph. A. p. 497. 1913.

KXAN , Z.F. Influence of period of vegetation and

development of plant on the alkaloidal content of
Hyoscyamus niger. J. A. Ph. A. p. 1163. Nov.
1931.

KOCH. G.P . Atropa Belladonna. J. A. Ph. A. p. 390.
1919.

. Cultivation of medicinal plants. J. A.

Ph. A. p. 275. 1919.

. Hyoscyamus niger. Am. J. Pharm. 11:68-83.
1919.

Yimrooijaia

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* ♦ • , . . .

, • r .ri •

0>' ^ rr

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,

r r .T fi

HO I Au 003 baa

,

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r ••• r

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64

16. _ . Distribution of mydriatic alkaloids in

Hyoscyamus niger . Am. J. Pharm. 91:68. 1919.

17. MEIXON and TI GEL AAR . Titration of certain alkaloids.

J. A. Ph. A. p. 676. July 1932.

18. MILLER, F.A. and MEADER , J.W. J. Ind. Eng. Chem.

5:1014 from B .Ph.C . 1914. 1913.

19. NEWCOMB and HAYNES. A.J.P. J XXXVIII:!. 1916.

20. PATU. Abstract from Pharm. Zentralh. LXIV:122.

21. SCHOU and BJERREGAARD. Dansk. Tids. Farm 6, 185-193.

22. SELF, P.A.W. Alkaloidal assays of B.P. 1932. Ph.

J. and Ph. p. 222. Mar. 1933.

23. S IE VERS, A.F. The germination of Belladonna seeds.

J.A.Ph.A. pp. 483-505. 1914.

24. _ . The distribution of alkaloids in the

Belladonna plant. Am. J. Pharm. 86:97-112. 1914.

25. . Belladonna alkaloids. A.J.P. 88:193.
1916.

26. WATKINS and PALKIN. Automatic devices for extraction

of alkaloids. J.A.Ph.A. p. 1099. 1925.

27. _ . The quantity of alkaloids in

Hyoscyamus and a new method for its evaluation.
J.A.Ph.A. p. 1039. Nov. 1927.

28. ANONYMOUS • Pacific Pharmacist 1909-1910, p. 295.

1909.

29. _ Abstract of proposed changes to U.S.P.X.

J.A.Ph.A. p. 1051. 1924.

30. _ . J. Pharm. 40:485. 1915.

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