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ORIGINAL COMMUNICATIONS

EIGHTH INTERNATIONAL

CONGRESS
OF APPLIED CHEMISTRY

Washington and New York
September 4 to 13, 1912

SECTION VIIIc: BROMATOLOGY

VOL. XVIII.

ORIGINAL COMMUNICATIONS
EIGHTH INTERNATIONAL

CONGRESS
OF APPLIED CHEMISTRY

Washington and New York
September 4 to 13, 1912

SECTION Vnic: BROMATOLOGY

VOL. xvni.

The matter contained in this volume is printed in exact accordance with the manuscript
submitted, as provided for in the rules governing papers and publioations.

La mati&re de ce volume a £t6 ImprimSe strlctement d'accord avec le manuscrit foumi et
les Fugles gouvernant tous les documents et publications.

Die in diesem Heft enthaltenen Beitr^e sind genau in Ubereinstimmung mit den una
unterbreiteten Manuskripten gedruckt, in Gem^ssheit der fuir Beitr&ge und Verlagsartikel
geltenden Bestimmungett.

La materia dl questo volume e stampata in accordo al manosoritto presentato ed in base
alle regole que governano i documenti e le publicazioni.

THE RUM FORD PRESS
CONCORD-N-H-U'S'A-

ORIGINAL COMMUNICATIONS

TO THE

EIGHTH INTERNATIONAL CONGRESS

APPLIED CHEMISTRY

APPROVED

BY THE

COMMITTEE ON PAPERS AND PUBLICATIONS

IRVING W. FAY, Chairman

T. LYNTON BRIGGS JOHN C. OLSEN

F. W. FRERICHS JOSEPH W. RICHARDS

A. C. LANGMUIR E. F. ROEBER

A. F. SEEKER

SECTION VIIIc. BROMATOLOGY.

Executive Committee.

President: W. D. Bigelow, Ph.D.

Vice-President: A. L. Winton, Ph.D.

Secretary: H. A. Baker, B.A.

Paul Rudnick

Chas. D. Woods, Sc.D.

Sectional Committee.

Charles S. Ash

E. H. S. Bailey, Ph.D.
H. E. Barnard, B.S.

F. W. Bedford, M.S.
Lucius P. Brown

R. E. DOOLITTLE, B.S.

Richard Fischer, Ph.D.
Elton Fulmee, M.A.
H. S. Grindley, Sc.D.
Rudolph Hirsch, B.Sc.
M. E. Jaffa, M.S.

C. Langley

C. F. Langworthy, Ph.D.
H. M. LooMis, B.S.
William McPherson, D.Sc,

Ph.D.
G. F. Mason, M.S.
L. H. Merrill, Sc.D.
Andrew S. Mitchell, Ph.C.
Harry Snyder, B.S.
John Phillips Street, M.S.
A. G. Woodman, B.S.

and the Sectional Executive Committee.

VOLUME XVIII
SECTION VIIIc: BROMATOLOGY

CONTENTS

Abb, Goro (see Takahashi, T.) ?*"=

AsBj Charles S.

The Relation of the Chemist to the Wine Industry 9

Ash, Charles S.

Interpretation of the Results of Wine Analysis 17

Baker, H. A.

Experiments on Feeding Guinea Pigs "Salts of Tin" in Measured
Quantities for Several Week» 31

Baker, H. A.

Special Adaptation of Iodine Titration Methods for the Estimation
of Tin, Especially in Connection with Determinations of "Salts
of Tin" in Canned Foods 35

Baker, H. A.

"Springers" in Canned Foods — Causes and Prevention 39

Baker, H. A.

Apparatus for Quantitative Extraction of the Gases in Canned
Food Containers 43

Baker, H. A.

The Disappearance of Oxygen in Canned Food Containers 45

Bartlett, J. M.

Eggs Preserved with Silicate of Soda 51

Bioelow, W. D.

Some of the Results of the Food and Drugs Act 57

Contents [vol.

BOEDAS, M. LB De. p^™

Sur I' analyse du Phosphore dans Us Cendres du Lait 65

BORDAS, M. LE Dr.

L'AcidiU originelle du Lait 67

BOEDAS, M. LE Dr.

De V action du Lait sur certains rSactifs 69

CtrsHMAN, Allekton S. and Fuller, H. C.

A Chemical Investigation of Asiatic Bice 73

De' Conno, Prof. Dr. E.

Sidla Maturazione Del Formaggio Pecorino 83

DUNNINGTON, F. P.

The Grinding of Corn-Meal for Bread 119

Frear, William

Food Standards, Their Nature, History and Functions 129

Fuller, H. C. (see Cushman, Allebton S.)

GooDNOw, E. H. (see Tolman, L. M.)

Hanson, H. H.

The Packing of American Sardines 131

Hebeet, M. Alexandee

Etude chimique des fruits de Sorindeia Oleosa 139

Hebeet, M. Alexandre

Sur la Composition de Divers Produits, Oraines ou Tubercules Amy-
lads ou Ficulents de I'Afrique Occidentale Frangaise 143

Ikeda, Kikunae

On the Taste of the Salt of Olutamic Add 147

Langwostht, C. F.

Progress Report of Nutrition Investigations in the United States 149

Langwortht, C. F. and Milner, R. D.

An Improved Form of Respiration Calorimeter for the Study of
Problems of Vegetable Physiology 229

LiNDET, M. L.

Sur le Role Antiseptique du Sel Marin et du Sucre dans les Conserves
Alimentaires 237

xviii] Contents vii

LooMis, H. M. Paoh

Salmon Canning Induatry of North America 239

LouHiE, H. L.

Proposed Method for the Estimation of Tin in Canned Foods . . . 247

MiLNER, R. D. (see Langwoetht, C. F.)

MUBAMATSU, S.

On the Preparation of "Natto" 251

Odake, S. (see Suzuki, U.)

Okttda, Y.

Contribution to the Chemistry of the Ripening of "Shiokara" 265

Okuda, Y.

Quantitative Determination of Creatine, Creatinine and Mono-
amino-adds in some Fishes, Mollusca and Crustacea 275

Olson, Geo. A.

The Effect of Modifying the Gluten Surrounding of Flour 283

Read, E. Alberta

A Method for the Detection of Color in Tea 301

Robin, M. Lucien

Recherche de Petites QuantitSs de Graisse de coco dans le Beurre de
V&che 305

RuDNiCK, Paul

The Chemist in the Service of the Packing House 309

Sawamura, S.

An Investigation on the Manufacture of Tea 313

Snyder, Harry

Wheat Flour. A Monograph 323

Stewart, A. W.

On Some Dried Milks and Patent Foods 329

Suzuki, U.

Uber die Chemische Zusammensetzung des "Salzbreies" von Bonito
{"Shiokara") 339

Takahashi, T. and Abe, Goro

On the Chemical Composition of "Saki" 349

viii Contents [vol.

TOLMAN, L. M. AND GOODNOW, E. H. P-^™

A Study of the Composition of Cider Vinegar Made by the Generor-
tor Process 359

WiNTON, A. L.

The Microscopical Examination of Vegetable Products as an
Adjunct to Their Chemical Analysis 361

YoNETAMA, C. (see Suzuki, U.)

THE RELATION OF THE CHEMIST TO THE WINE

INDUSTRY

Charles S. Ash
San Francisco, Cal.

I think that I am correct when I state that there are few, if
any, industries where it is so difficult for the chemist to prove
his necessity as in the manufacture of wine. This is due to the
following facts:

The most common difficulty is that people connected with the
wine industry have absolutely no idea of chemical science and
cannot see any use for the chemist in their business.

The wine industry is not what we would call a true manu-
facturing industry, as for example, the sugar industry; therefore,
the chemist has no value in a mathematical capacity. He does
not trace out losses in manufacture as does the sugar chemist.
It is apparent to any business man that if there are 10,000 lbs.
of raw sugar put into a refinery in a day and if 9,500 lbs. are
recovered in the refined product that he has a loss of 5 per cent.
The chemist here is of value to him to ascertain definitely the
loss of manufacture, to help him minimize that loss, trace up
leaks, improve processes of refining and many other incidentals,
which has made the sugar chemist renowned throughout the
world. This example holds good in any manufacturing process
where the raw material is converted into a finished article. Even
in the kindred industry of grain distillation, we have the obvious
necessity for the chemist; for it is a self-evident fact to the dis-
tiller that the man, who can increase the jaeld of alcohol from
a given weight of cereal, is of value to him. In the yeast industry
(another kindred industry) this example holds good. In
fact an instance has come to my knowledge where a chemist
has increased the yield of yeast for his employers over 30 per
cent. All of these things mean profit to the merchant and
profit is "Raison d'etre" of all enterprise. In other manufac-
turing industries, the chemist has proved his value by making
2 9

10 Original Communications: Eighth International [vol.

immense fortunes in by-products. In a strictly chemical line,
as in the manufacture of chemicals, as well as in the manufacture
of dyes and colors, the whole industry is absolutely dependent
upon chemical laws hidden or obvious. The innumerable new
dyes and colorings are the product of brains alone and would
never have seen the light of day but for the master minds, who
have built up these synthetical products. As grapes are
very poor in by-products, and as wine is a natural product, the
wine chemist was shut out from usefulness in this direction.

Compared to the above industries, the wine industry is the
most venerable; wine having been made by the Egyptians many
centuries before Christ. It is mentioned throughout the Old
Testament repeatedly. As wine has made itself spontaneously
long before such a thing as chemistry was ever dreamt of (that
is our conception of exact chemical science) it was considered
a natural product. About fifty or sixty years ago, when Pasteur
made his classical researches, considerable light was thrown on
the subject and for the next fifty years, his views made them-
selves slowly felt throughout the world. The first effect was in
the beer industry where yeast was planted in sterile medium and
the quality of the beer has depended greatly upon the quality of
the yeast. In the wine industry this was not the case, as the
process of making wine was roughly this : The grapes were crushed
and allowed to ferment. They fermented spontaneously on their
own yeast and made good or bad wine as the composition of
the grape, chmate, temperature during the fermenting period
and other conditions allowed. We hear to this day the most
abused phrase of "the especially fine vintage of 188 — ". This
was simply that the grapes ripened well and that the temperature
during the fermenting period was ideal. Such a thing as con-
trolling the temperature was unthought and unheard of.

When the chemist looked for employment in the wine industry,
he was asked what he could possibly do, what he could possibly
find out that an expert taster did not know. How could he
benefit the wine industry in any way? He replied that he could
analyze the wines, determine the amount of alcohol, total acids,
volatile acids, solids, tannin, glycerol, etc. This brought the
reply, "Well, if I did know, what good is it to me? I am glad

xvin] Congress of Applied Chemistry 11

to know that wine has, for example, 12 per cent alcohol, and has
a total acid of 6 parts per thousand and volatile acid of one part
per thousand but, after I do know it, what does it mean to me?
I am very pleased to know it but it does not do me any good.
It does not tell me whether the wine is good, bad or indifferent;
I can tell from my taste all those things. There is nothing I
cannot tell. I have been running this wine business for the last
fifty years without a chemist and made money, and I should be
able to run it a great many more years without one." What
he said was, in a way, absolutely true. This was the unfortu-
nate thing because a chemist, applying for a position in the wine
industry, is not a wine man and knows nothing about the nature
of wine. His value only starts to be of importance when he does
become a wine man. The fallacy of the wine merchant at this
time (fifteen or so years ago) was that he did depend on the
chemist, that, in fact, all over the world preservatives were used;
a chemical product itself, preservative values having been found
by chemists. Therefore, no matter how troublesome a wine
may have been, preservatives in a great or small quantity were
added, and the wine kept indefinitely. Chemists, however,
were finally employed in the wine industry. I think that the
fijst wine laboratory of any corporation in the United States,
and possibly one of the first in the world, was formed in 1895;
Before long the chemist was able to tell the merchant what 12
per cent alcohol meant, what the total acidity of .5 per cent
meant and what 2 J per cent of solids meant; and, furthermore,
what the expert taster could taste, what he could not taste and
could also prove the taster was correct or incorrect in his opinion
of a wine. For example, if he considered a wine sound that had
.150 grams volatile acid (as acetic) per 100 c.c; then the taster
was wrong, that the wine was sour that contained such a high
percentage of volatile acid. This soon became apparent to the
taster himself and a new crop of difficulties arose. He wanted
to be checked in his work and expected a determination of vol-
atile acid (or as we commonly call it, "volatile") could be done
almost as fast as he could taste wine and if he had 200 or 300
samples, he would expect them to be done in one day. He
wanted them running day and night, Sundays and hohdays, not,

12 Original Communications: Eighth International [vol.

of course because they were of any value but simply to prove his
opinion and the poor, lonely wine chemist, harassed as he was,
and upon trial, had very little satisfaction in finding one test, be-
sides that of alcohol, which was of some value to his employer.
But now other difficulties arose for the wine merchant. Pure
Food Laws were coming into effect in European countries. Sal-
icylic Acid (the common preservative used) had to be abandoned
and the chemist was instructed to look for a preservative that
could not be detected (and he is still looking). Then benzoic
acid was adopted as a preservative, as benzoates, at that time,
were very difficult of detection. Chemical science soon, however,
caught up to them so that they were of no value. However, dur-
ing this period, the chemist had become acquainted with the wine
business and learned a few relations between composition of the
wine to its keeping qualities and, therefore, was able to interpret
an analysis. He could anticipate, to a certain extent, whether or
not a wine would blow up, turn soiur or spoil in some way when
it reached a warm climate and, instead of finding a new preser-
vative, concentrated his thoughts on getting the wine into such a
condition that there would be no need of a preservative, and
that a preservative would simply increase the cost of the wine
and do it little, if any, good. This I think holds good in all food
industries; that the need for preservatives decreases as our
knowledge of the product increases. Through the efforts of the
writer, wine was shipped without preservatives as an experiment.
The first car was shipped out with many misgivings. It was
expected that every barrel would blow up, and the entire car
be lost. This was a very trying time for the chemist, as his
theories were on trial and a failure meant a return to the old
regime. The car, however, gave perfect satisfaction and nothing
was heard afterwards. From that time on, the amount of pre-
servatives decreased and methods for improving the vintage
increased; in fact, a chemist's knowledge took the place of pre-
servatives in this business. This was sometime before our
national Pure Food Law. The firms, not employing a chemist,
were found to be at quite a disadvantage, as their goods with
preservatives, were subject to seizure, both by municipal and
state authorities. This gave them undue notoriety and they

xviii] Congress of Applied Chemistry 13

were forced to hire a chemist to improve and control their methods
of manufacture and preparation of wine for the market. So much
for the troubles and difficulties of pioneer chemists in the wine
industry.

The chemist, having proved his value, now reached out to
other things. He now started to look into the many causes of
trouble. First, why some wines would not clarify, why wines of
some localities and some districts degenerated, why some wines,
shipped from the cellar in excellent condition, spoilt. These
were the problems that confronted and still confront the wine
chemist. He found out, however, that wines would not clarify,
usually either on account of disease or on account of the com-
position of the wine, usually insufficient tannin. Then he started
to improve the clarifying medium and to look into the reason
of the degeneracy of wine after shipment. He found certain
diseases which had to be, and were, overcome (these diseases,
of course, never took place when preservatives were used, as the
antiseptic action was too great for the micro-organisms to over-
come). He then started to look into reasons of locality to find
out why some cellars and some vintages turned out very poor
wine. This, of course, was not original in California alone, as
it was found in every other place throughout the world, and a
great deal of literature on the subject was being printed so, while
this did not take a great amount of original research, he acted
as a medium to distribute scientific knowledge to the cellar super-
intendents, so that their methods of handling the vintage would
be improved. This, also, at times, meant an outlay of money
and is always, in corporations, a difficult thing to obtain. It
was proven that wines, fermented at a high temperature, spoilt
while those, fermented at a low temperature, nearly always kept
sound. His labors then took on methods of controlling the
temperature of the vintage, improving and handling, and also
to ferment on cultivated yeasts of known virility and not leave
the fermentation to chance. The quality of the wine, as well
as its keeping properties were improved and the amount of spoilt
wine reduced to a minimum. It is apparent that a successful
business must turn out a uniform product. There is no difficulty
in turning out a uniform sugar for example, but with wine, every

14 Original Communications: Eighth International [vol.

vintage is slightly different and, if a brand of wine is established,
it is necessary to supply your customer with wine of exactly the
same type. Otherwise, the consumer, being used to one wine
objects to any other. The wine chemist has to help in the
production of uniform products. As the blending of wines is
the final operation (and as these blends compose at times 100
different wines) this is perhaps one of the most important duties
of a wine chemist. Blends are usually made up in sample,
analyzed, blended as near as possible to the composition of the
previous blend of this type, and the blends are then distributed
to the winery, which is to make them up and, after blending, are
sent back to the laboratory where the chemist analyzes them again
to check up and see whether these blends have been properly and
uniformly made.

The following samples show the method of checking the
blending. The analyses of the sample blend (made in the lab-
oratory) and the actual blend (made in the cellar) must agree,
otherwise, the blend is not uniform and must be reblended :

Per cent
Alcohol by

Grams

per 100 Cubic Centimeters

Volume

Total
Acidity

VolatUe
Acidity

Reducing
Sugar

Ta,Tinin

Winehaven Claret

Blend No. 485

(265,000 gal-

ena).

Sample Blend . . .

12.29

.500

.060

.145

.135

Finished Blend . .

12.37

.510

.060

.150

.140

Wahtoke Port

Blend No. ^82

(144,500 gal-

lons).

Sample Blend . . .

20.78

.390

.043

6.60

.070

Finished Blend . .

20.78

.390

.046

6.63

.079

Besides this the wine chemist has duties in common with all
chemists. He must analyze the water and soils of all the vine-
yards owned by his company, analyze the suppUes, such as used

xviii] Congress of Applied Chemistry 15

either in wines or in the vineyards, advise as to fertilizers to be
used and devise means to gather as many by-products as possible.
I may say in conclusion that the wine chemist, in spite of
temporary discouragements, is having more intimate relations
with the Wine Industry. He is, in fact, becoming quite friendly,
and he has hopes of being on the same good terms as his brothers
in the sugar, dyeing, oil, petrolemn, gas, soap and other industries,
which the chemist has made famous.

INTERPRETATION OF THE RESULTS OF WINE
ANALYSIS

By Charles S. Ash
San Francisco, California
Introduction.

It is evident that the analysis of any product is useless unless
it can be correctly interpreted; in other words, every analysis
that is made must be interpreted to be of any value. What
the diagnosis is to the physician, interpretation of analytical
data is to the chemist. Some of these interpretations are purely
mathematical. The value of sugar cane is dependent on sugar
content; ores are valued by the amoimt of metal they contain,
and, therefore, assays are interpreted with little or no diflB-
culty. The interpretation of analytical data on some other
products are, on the other hand, most diflScult and it is only by
a careful study of many analyses of these products of known
origin that we are able to show the true meaning of their chemi-
cal composition. In food products, we are quite content, as a
rule, to tell from analytical data whether the food in question
is pure or adulterated. As this interests most chemists, we will
confine ourselves almost entirely to this question.

The interpretation of wine analysis has been confined almost
entirely to the judgment of its purity; in fact, the only result
of these interpretations has been the formation of a set of stand-
ards to which a wine must conform in composition to be con-
sidered pure wine. These hard and fixed standards do not
accomplish the object of detecting adulteration on one hand
while, on the other, they often work real hardship on wines of
pure origin. Grapes of the same variety, grown in different soils
and in different climates, produce wines of absolutely different
taste, and composition. If this is true of the same variety of
grapes, under different conditions, what must be the difference
in composition of himdreds of varieties of grapes grown in
almost every condition of soil and climate in the world! This
we will discuss later.

18 Original Communications: Eighth International [vol.

There has been a tendency in the past, in writing of interpre-
tations of wine analysis, to quote the interpretations of previous
authors and this has influenced our already meager literature
on this subject to such an extent that we have had but little
original thought for many years, the last interpretation being
only a compilation of previous data. To avoid such a tendency,
the present writer will avoid previous literature on the subject,
preferring to treat it from an independent viewpoint. I thjnk
you will pardon the writer for his temerity in taking this stand
when he explains that yearly, for the past fourteen years, there
has been received in his laboratory from 15,000 to 30,000 sam-
ples of wine of known origin. All these wines are examined and,
at least, one-half analyzed.

We will, therefore, consider that the interpretation of wine
analysis has two objects, — one to judge the purity of wine and
the other to judge its quality and condition. We will take,
under these two heads, part 1, dealing with adulteration and
part 2, with condition, soundness and disease.

Part 1. — The object of all adulteration is to cheapen the
article in question or to increase its commercial value by artifi-
cial and false means. No one will adulterate unless it is profit-
able and no one will substitute an artificial product, which costs
more than the natural one. This, of course, is plain. In wine
then, as in every other product, adulteration aims to decrease the
cost of production or to increase its selling value. We, therefore,
will consider the various forms of adulteration which may be used.

Addition of water

1. — Increase of volume Artificial wines

Wines of foreign fruits
Addition of spirits and
sugar, or both

3. — Increase of stability I Preservatives

^. — Improvement of appearance . . . { Artificial coloring

2.— Increase of strength.

Artificial flavor, saccha-
rine, etc.

5. — Chan ging of taste

6. — Modified or fixed spoilt wine.

7.— To these forms of premeditated adulteration, we may still

have adulteration which will come imder the head of

XVIIl]

Congress of Applied Chemistry

accidental adulteration. Under this head, will come the
presence of heavy metals, as zinc, iron, copper or arsenic.

Dilution.

Of all forms of adulteration, the most common, the most
profitable and the most difficult of detection is dilution: the
simple addition of water. The first two facts are self-evident.
Why dilution is so difficult of detection needs some study.
Having already touched on the effect of climate, soil and variety
on the composition of the grape, we will go into further details.
Now, it hardly seems necessary to state that grapes, grown in
warm climates or grapes ripened to perfection, have more
sugar than unripened grapes. It is also obvious that grapes
that are unripe have a greater acid content than ripe grapes.

The following example shows the effect of climate on the
composition of the juice of the same variety of grape:

Variety Carignan

(1)

(2)

Grown in France (Midi)

Grown in Fresno, Cal.

Density at 60F

1.076
18.81
16.2
.840

.0997

Total Solids, Grams per 100 c.c.
Reducing Sugar " " " "
Total Acids as Tartaric " "

24.2
23.26
.590

Potassium Bi-Tartrate " "

.580

.376

The next example illustrates the effect of ripening on the

composition of the grape juice:

(3)
Variety Zinfandel

Sugar (% Balling)

Acidity as Tartaric

July 10
" 20

6
9

1.12
1.10

Aug. 1
" 15

14
20

.980
.700

" 20

.620

20 Original Communications: Eighth International [vol.

Now, the essential point I wish to bring out is this: that
grapes, high in sugar content must necessarily be low in acid
content and vice versa. Both conditions, that is, high sugar
content and high acid content rarely exist in the same grape.
On the other hand, low sugar content and low acidity also do
not exist in the same grape. This is a generality. This is very
important. It will, therefore, be seen that the resulting wine
from grapes of high sugar content, will be high in alcohol and
low in acidity while wines, made from grapes low in sugar con-
tent, would give a wine low in alcohol and high in acidity. To
repeat again, both high alcohol in a wine and high acidity, and
low alcohol and low acidity, do not exist. We have, therefore,
in cold countries the difficulty of obtaining grapes that ripen to
a sufficient sweetness so as to give wine of high enough alcohol
to preserve itself and a low enough acidity to be drinkable while,
on the other hand, in warm regions, we have the reverse trouble
of getting wines of high enough acidity and low enough in alcohol
to have sufficient character and flavor to be considered desir-
able wine. These wines, as you see, are pure wines of absolute
different composition and a hard and fixed standard made in
either country to suit the conditions of the native wine may
work hardship on the wines of the other country or, on the other
hand, if the standard for example, happens to be based on the
composition of the wine of the cold country, there would be
little difficulty in diluting the wine from the warm country and
still conform to the standard of pure wine.

I have spoken on the difficulty of a fixed standard. Let ite
take, for example, the American standard. This standard was
meant to be, and is, a liberal standard, which tries to embrace
all pure wines of the world. This standard is as follows:

Red

White

Alcohol % by volume

7 to 16
.14

.1%

.16

Not less than 1.60

" more " .10

tt tt tt 20

7 to 16

Volatile Acid as Acetic, Grama
Reducing Sugar "
Ash

per 100 CO

tc CC it

.12

1%
.16

Sugar Free SoUds "
Sodium Chloride
Potassium Sulphate "

it tt it
tt tt it
It (1 ((

1.40
.10
.20

xviii] Congress of Applied Chemistry 21

Let us say, for example, we have below an example of aver-
age, normal California Red Wine with the following composition:

Alcohol % by Volume 12%

Volatile Acid Grams per 100 cc 100

Sugar Free Solids 2.50

Ash 270

Sodium Chloride 005

Potassium Sulphate 015

This wine can safely be diluted one-half (2-3rds wine and l-3rd
water) and still be able to conform to these standards. The
composition of the diluted wine would then be as follows:

Alcohol % by Volume 8%

Volatile Acid, Grams per 100 cc . 067

Sugar Free Solids 1-67

Ash 180

This comes well within the standard of purity as laid down by
our government. On the other hand, these standards will work
hardship on some of the finest old wines produced. This we will
discuss when we take up the question of Volatile Acid of the
wine, as it is out of place under this heading.

To return ; we have shown what can be done imder the stand-
ards laid down by our government. There is nothing in these
standards showing the relation of alcohol to total acidity, much
less showing the composition of this acidity in natural wine.

In this case cited, the original acidity of the imdiluted wine
was .6 grams per hundred cubic centimeters. When this wine
is diluted, we find an acidity of .4 grams per hundred cubic
centimeters. A natural wine with an acidity of .4- grams per
100 cc. and an alcoholic strength of 8% hy volume does not exist.

The obvious question is then asked: why a diluted wine can-
not be acidified to raise the acidity to that of normal wine of
such an alcoholic strength. This certainly complicates matters.
The only available acids are, as we know. Citric and Tartaric
Acids. As Citric Acid is never present in grapes in quantitative
amounts, its presence in most cases will be indicative of manipu-

22 Original Communications: Eighth International [vol.

lation. We have now only Tartaric Acid. The natural non-
volatile acidity (fixed acidity) of wine is largely made up of
Potassium Bi-tartrate and not Tartaric Acid. Free Tartaric
Acid is only present in small quantities in natural wine. So, if
Tartaric Acid was added to this diluted wine we will have a
larger amount of free Tartaric Acid than Potassium Bi-Tartrate.
We cannot use Bi-Tartrate to acidify the diluted wine, as it is
only sparingly soluble. The following table is explanatory:

Natural

DiltUed

DUuUd

Wine

Wine
Acidified

Alcohol % by volume

12.0

8.0

Total Acidity (as Tartaric) Grams per

100 CO.

.600

.400

.800

Volatile " (as Acetic)

.100

.067

Sugar Free Solids

2.50

1.67

Ash

.270

.180

Potassium Bi-Taxtrate " "

.300

.200

.2000

Free Tartaric Acid " "

Trace

.400

Here, in the natural wine, we have .3 grams Potassium Bi-
Tartrate and a trace of Free Tartaric Acid, and in the diluted
wine, we have .2 grams Bi-Tartrate, while in the diluted acidi-
fied wine, we have .2 grams Bi-Tartrate and .4 grams Free
Tartaric Acid; the Free Tartaric Acid, being in excess of the
Bi-Tartrate, while it should only be one-third at the most.
Now, suppose we add the alcohol % by volimie and the total
acidity (grams per litre). We have in the natural wine a total
of 18, in the diluted wine 12, and in the acidified wine 16.
Whenever a Red Wine has a total acidity of less than 16, it
should be investigated. All this data is relative to the effect of
dilution on acidity. I know the difficulty of showing every-
thing in one example, but we have already gone to greater
length than I had wished to go. It is obvious to us all that
dilution will lower the percentage of soHds, the percentage of
ash and other constituents of the wine. I might state that the
alkaUnity of ash, figured in terms of Bi-Tartrate should ap-
proximately equal the percentage of Bi-Tartrate found in the

xviii] Congress of Applied Chemistry 23

original wine; in other words, the alkalinity is almost due
entirely to the Bi-Tartrate which is converted into Potassium
Bi-Carbonate by incineration. In conclusion, it would appear
that California red wines, having a sugar free solids less than
2.30 in conjimction with an alcohol plus acid total of less than
16, is to be regarded with suspicion. With white wines, a sugar
free solids of less than 1.60 and alcohol plus acid total of less
than 15, should also be looked upon with suspicion. Wines,
having a large amount of Free Tartaric Acid, in proportion to
the Potassium Bi-Tartrate, and coupled with low sugar free
solids would indicate that the wine had been diluted and acidified.
The relation of Potassium Bi-Tartrate to the free Tartaric Acid
is the same in white wines as it is in reds.

Artificial Wines

These wines are made by fermenting sugar, sucrose or glucose,
either alone or thrown over grape pomace. They are, in conse-
quence, very light in color or colorless, and have either to be
blended with natural wines or colored artificially. They are
also high in free Tartaric Acid and low in Bi-Tartrate and low in
solids and ash. They often have no Potassium Bi-Tartrate at
all, the acidity being due to free Citric or Tartaric Acid. Those
made from glucose are always high in solids, due to dextrin or
other unfermentable substances. Such substances are too easy
of detection to be mentioned here. Those made from sucrose
are always a superior article. They are, however, very expen-
sive and in countries where grapes are cheap (as in California)
cost more than the natural wine. For example, sucrose cost, say,
5c per lb.; grapes, having a sugar percentage of 22 per cent
will have 440 lbs. of sugar per ton of grapes; 440 lbs. of sugar,
in turn, at 5c. per lb. would cost $22 and, therefore, sugar would
be equal to grapes costing $22 per ton. The average price
of grapes in CaUfornia, year in and year out, will be from $12
to $30, the average cost being somewhere around $18. Now,
it is obvious that no one would use this form of adulteration
imless the price of grapes were well over $30 per ton. In colder
countries where the grapes do not ripen to a sufficient sugar

24 Original Communications: Eighth International [vol.

percentage, the sucrose is very often added to give the resulting
wine sufficient alcohol to preserve it. When this is not done to
excess, it is almost impossible (without a thorough knowledge of
the composition of natural wines of such district) to show this
form of adulteration. Sucrose is inverted by the action of the
yeast and acid, before fermentation. The inverted sugar is very
closely alUed to that of the natural sugar of the grape. I might
say in conclusion, all the artificial wines, that the writer has
seen, have come, of course from localities where grapes were
dear. Their acidity has been due almost entirely to free Tar-
taric or Citric Acid. They are often artificially colored and
preserved with some preservative. Most of these have been
made from glucose. Sometimes sweet wines, like Port, have
been made. These wines contain about 12 per cent alcohol and
as high as 20 per cent of total solids, the method of making
being that the glucose solution has fermented imtil fermentation
takes place no longer and the wines stick. Preservatives are
then added and the wine is colored; sometimes synthetical
flavors are also added. These wines never cloud and give no
trouble, during changes of temperature in climates, that all
natural wines do.

FoBEiGN Fruit

This form of adulteration I hardly think exists in the state of
CaUfornia on account of the low price of grapes and the com-
paratively high price of other fruits. In Europe, apples and
figs have been, at times, fermented and mixed with natural
wines. Such wines, by themselves, would easily be detected,
as very few other fruits, beside the grape, contain any Bi-
Tartrate. The absence of Bi-Tartrate or a very low percentage
of same in a comparatively normal composition otherwise,
would tend to show this form of adulteration. When more light
is shown on the acid composition of various fruits, no trouble
will be experienced in detecting such adulteration.

Increase of Strength

This is done by the addition of sugar, as had already been
described, or spirits, or possibly both. The addition of spirits

xviii] Congress of Applied Chemistry 25

cannot take place in American dry wines, as wines must have
4 per cent of total solids to be eligible to fortification. Neither
can taxpaid spirits be added to such wines, for it is against the
revenue laws to mix taxpaid and free spirits. Such a form of
adulteration would be criminal and liable to prosecution. Some
countries allow spirits to be added to wines for exportation.
This is readily detected on account of its abnormally high
alcohol together with high acidities. The addition of spirits
and then dilution by water afterwards, would make a double
dilution and would be detected under means already described
in the paragraph on dilution.

Increase of Stability

Whenever wines have been diluted or made from unsound
material, preservatives are sometimes added to give them
keeping qualities, without which they would get progressively
worse, so as to be impossible to market them. In this connec-
tion the use of preservatives, irrespective of whether they are
harmful to the human system or not, should be prohibited in the
wines for the reason that poor or putrid articles could be mar-
keted, which would be impossible to do without their use.

Improvement of Appearance

Whenever artificial or imitation wines are sold, either alone or
mixed with natural wines, the color is insufficient to meet the
popular demand. It is, therefore, imperative that these wines
be given the appearance of normal wines. To do this, they must
be colored. Aniline colors are used for this purpose almost en-
tirely, as vegetable colorings are not fast and either fade or
deposit when subjected to daylight. The usual clarifying
methods (the addition of albumen) will very often deposit
vegetable colors and, therefore, after wine has received a clari-
fication, vegetable colorings are almost entirely removed from
the wine. The above statement, in reference to the coloring of
wines, is applicable to red wines entirely. White wines are
rarely colored, though in isolated cases, the writer has seen
wines given green tints by artificial means.

26 Original Communications: Eighth International [vol.

Change of Taste

Sometimes flavors are added to natural wines to improve the
state, or give them either the appearance or taste of old wines
but, in most cases, this is a waste of money. Such form of
adulteration is very difficult of detection. However, a higher
percentage of esters than is normal in a wine, especially volatile
esters, will tend to show adulteration of this form. Saccharine
has been at times added to white wines to imitate wines of the
Sauterne type. This is very simple of detection, 1st — by the
method of detecting saccharine itself, and, 2nd., that the total
soUds of such a sweetened wine is no higher than the normal
solids of a dry white wine. Such sweetened wines are also
very low in reducing sugar and, therefore, cannot derive their
sweetness from natural sources.

Modified ob Fixed Spoilt Wines

This is the last form of premeditated adulteration. Spoilt
wines or sour wines, which have either been made from spoilt
grapes or have been spoilt by neglect or degenerated by some
disease, are often marketed after manipulation. The common
method of doing this is to neutralize the excess of acidity, either
by potassium, calcium or magnesium salts — usually calcium
carbonate is used to do this work. The resulting ash of such
wines is very high in hme salts, the total ash going considerably
over 1-lOth of the sugar free extract. The fixed acidity is usu-
ally very low in comparison to the volatile acidity. This is due
to the fact that such manipulation is rarely, if ever, successful,
and wines, having been neutrahzed in this way, usually are only
temporarily checked; the disease continues and more volatile-
acid is generated; the neutralization only neutralizing the acids
present at the time of neutralization.

We now come to the final form of adulteration, which we
will call Accidental Adulteration. That is the presence of
small amounts of heavy metals in wine. The presence of cop-
per, tin or zinc is due to the pipe line, which the wine is run
through. The amounts are very low indeed and it is almost im-
possible to avoid these contaminations. Carelessness, however.

xviii] Congress of Applied Chemistry 27

in allowing wines to stand in pipes until they corrode will very
often increase this small percentage to an enormous amount.
The presence of arsenic in wine can rarely be considered acci-
dental adulteration, as it is a natural constituent of grapes in
some localities. The largest amount ever seen by the writer in
California wines is one part in four million: usually one part
in twenty to fifty million is normal. Arsenic occasionally
is added to the wines, by the use of sulphur in fumigating
casks. This, of course, would be considered accidental adultera-
tion.

I wish to conclude, as I have begun, by wishing to avoid a
fixed standard applicable to all wines in the world. There is
only one way which we can tell with approximate certainty
whether a wine is pure or adulterated and this is to have for our
standard of pure wine the same standard as the country from
which the wine originates; for example, standards set down by
the Swiss government for Swiss wines, should be applicable to
Swiss wines; those set down by the German government for
their Rhine and Moselle wines, and other districts, should be
applicable to wines of that district; for French wines, those set
down by the French government as standards for their natural
wines of different districts, should be used on such wines. For
American wines, standards should be made by the United States
government or by the Pure Food Authorities of the different
states where the wine is grown. In this way, we will not demand
the same composition for Algerian wines as we do for German
wines, nor the same for Swiss wines as we do for California
wines. This I believe will be a very simple matter and, in my
opinion, is the only way of controUing this question. A unifi-
cation of analytical methods would also be a big factor, and
should be urged by such bodies as this Congress. If not, we
must follow the analytical methods of the country in question,
that is, if we are examining French wines, we must realize
that these standards have been based upon methods of official
French analysis, and results, using any other methods, will
either work hardship or may defeat the enforcement of a Pure
Food Law.

28 Original Communications: Eighth International [vol.

Under the Head of Soundness Part 2.

Just one word, before I close this already too lengthy paper,
in regard to unsound and diseased wines. Such wines can be
made from grapes and be pm-e in every way and still be imfit
for human consiunption. A parallel case would be in any putrid,
decayed or deteriorated food product.

This may be due to the following reasons, as the manufacture
of wine from rotten or diseased grapes, or the wines may be
made from soimd grapes, and become diseased from improper
methods of fermentation or neglect. To be brief, we will sup-
pose that the Must contains

Sugar 20 grams per 100 c.c.

Acidity as Tartaric 8-10 grams per 100 c.c.

The corresponding wine, fermented imder normal conditions,
would have a composition somewhat as follows :

Alcohol 11.5 per cent by volume

Total Acid 600 per cent

Volatile Acid 060 per cent

Reducing Sugar 120 per cent

However, if this wine should be fermented badly, or the tem-
perature rise too high, so that the yeast is either killed or Has
dormant, secondary fermentation will set in, and we will have a
wine of quite different composition, depending on the severity
of conditions or neglect in handling. A typical composition of
such a wine would be as follows :

Alcohol 11 per cent by Volume

Total Acid 700 per cent

Volatile Acid 200 per cent

Reducing Sugar 500 per cent.

Notice the difference of these two results made from the same
Must. Let us go backward and say that we have a spoilt wine
with this composition and see what we can tell about it from
analytical data. In the first place, notice that this wine has a
high volatile acid. This shows that it has been attacked by

xvni]

Congress of Applied Chemistry

micro-organisms and that secondary fermentation, in conse-
quence, has set in. The product of such fermentation (the high
volatile acid) is present. We then look at the reducing sugar and
find that it is high. We see that the conversion of sugar into
alcohol has not been completed and this confirms our opinion of
secondary fermentation. The wine is absolutely spoilt and
should not be used imder any conditions. This wine should not be
blended with any other wine, as it would simply contaminate the
entire blend. A great many mistakes are being made in trying
to work off wine of this kind in small quantities. This is an
absolutely bad practice, for nine times out of ten, the entire
blend is ruined. It is always best to make your first loss at once.
Wine of this type, of course, would make excellent wine vinegar,
and could be used for such purpose. In well-controlled wineries,
wine of this type is reduced to a minimum

We will now follow new, sound wine over a period of years and
see what changes we may expect in the volatile acid content.

Dry Red or
White

Sherry

Port, Angel-
ica Muscat,
etc.

Sound new wine

Sound wine 1 year old . . .
Sound wine 3 years old .

it (t e tl it

" " 8 " "
" " 10 " or

.05 to .08

.04 to .08

03 to .05

06
.09
.100

120 '

140

.100
.120
.140
.160
.170

.06
.08
.100
.120
140

.08

.100

.120

.160

.180

.04
.05
.07
.08
.100

.06

.07

.09

.100

.120

This represents the natural increase of acidity one would
expect. This is not intended as a fixed standard, but rather a
guide. From this it is seen that a yoimg wine, which would be
condemned as spoilt, may be a perfectly fine sound old wine.

EXPERIMENTS ON FEEDING GUINEA PIGS "SALTS
OF TIN" IN MEASURED QUANTITIES
FOR SEVERAL WEEKS

By H. a. Baker

New York

Seven normal young Guinea Pigs, of average weight of about
257 grams, were put in separate pens and was each fed daily,
except Sundays, one Gelatin Capsule containing 12.6 milligrams
of tin in the form of Hydrates mixed with Corn Meal.

The capsules were prepared in the following way:

Four grams of pure tin were dissolved in a small amount of
Hydrochloric Acid and evaporated almost to dryness. This was
then neutralized with Sodium Carbonate and dried, after which
it was mixed with fine Corn Meal. 55,000 miUigrams of the
mixture contained 4000 milligrams of tin.

201 Gelatin Capsules, weighing 11.438 grams, were filled with
this Corn Meal mixture and found to weigh 46.281 grams, so
that each full capsule weighed 230.25 milligrams. Subtracting
the weight of the empty capsule, 56.9 milligrams, we have as the
contents of each capsule 173.35 milligrams of Corn Meal mix-
ture, which contained 12.607 milligrams of tin.

These capsules were all standard size so that we consider the
amount of tin in each capsule to be the same, especially since
the amount of Tin Salts in the mixture was small.

The capsule was administered to the Guinea Pig by forcing
it down his throat, in order to make sure that the proper dose
had been given.

The Guinea Pigs were given an ordinary diet of Carrots, Let-
tuce and Crackers.

As will be observed from the table appended, the Guinea Pigs
showed different resistance toward this chemical. Five of the
Guinea Pigs were fed until death occurred. They lived the fol-
lowing number of days: 9, 10, 19, 19, 22, respectively. Two

32 Original Communications: Eighth International [vol.

other Guinea Pigs which had taken a capsule a day for twenty-
five days lived, but were after that fed no more capsules.
Twenty-three days after these two Guinea Pigs had been fed
no more capsules, they suddenly died from exposure from an open
window.

In addition to observing the doses which were fatal to these
Guinea Pigs, their livers were analysed in order to see whether
or not any tin had become stored up there. Examination showed
the livers of the dead Guinea Pigs to be somewhat bleached on
the outer edges.

It can be observed from the table that these Guinea Pigs accum-
ulated in their livers a small amount of tin each day, as long
as the dose was administered. This rate of accumulation was
about ^ of a milhgram of tin per day; the average rate for
five Guinea Pigs being .113 milligrams of tin per day.

Using this rate as a basis for calculating the amount of tin in
the livers of the Guinea Pigs numbers six and seven, at the end
of the twenty-fifth day, we would have 2.827 milligrams of tin
present in each liver. At the end of the twenty-third day, dur-
ing which period no more tin had been fed, we still found .3 and
.5 milligrams of tin in the livers of Guinea Pigs numbered six and
seven respectively, which means that the tin had been eliminated
from their livers at the average rate of .1099 milhgrams and
.1011 milligrams per day respectively. From the experiment, of
course, it cannot be known that this average rate of elimination
was the actual rate, and it probably was not. It is probable
that the rate of elimination varied according to the concentra-
tion of the amount of tin in the livers of the Guinea Pigs and
fell off as the concentration decreased.

It may be observed from the table that the average absorp-
tion in the livers of the amount of tin fed to the Guinea Pigs was
.898%.

These individual doses were very large, and no other experi-
ments have been carried on to determine what the effect of
smaller doses for longer periods of time would have been.

The exceedingly small amounts of tin involved in the deter-
minations were estimated by a very delicate iodimetric titra-
tion method, using N-lOO Iodine. We consider the accuracy of

XVIll]

Congress of Applied Chemistry

this titration to be ■within 3-10 of a cc, which means that the
determinations are within .2 milligrams of accuracy, probably
always tending somewhat toward high results.

TABLE SHOWING RESULTS AND ANALYSES FROM EXPERIMENTS
ON FEEDING GUINEA PIGS "SALTS OF TIN" IN MEASURED
QUANTITIES

By H. a. Baker

GUINEA PIGS

No. 1

No. 2

No. 3

No. 4

No. 5

No. 6

No. 7

No. days fed Salts of Tin

Amount in Milligrams of

tin as "Salts of Tin "fed

12.6

113.4

Death

12.7

260
4.88
1.05

.1166
.926

12.6
126.0
Death
9.8

215
4.56
1.015

.1015
.805

12.6
239.4
Death

10.0

251
3.98
2.30

.1210
.961

12.6
239.4
Death
8.9

298
2.99
1.80

.0947
.752

12.6
277.2
Death

10.0

306
3.27
2.90

.1318
1.045

12.6

315.0

Living

9.5

(Detei
23 day

248.5

3.82

25
12.6

Total Tin fed as "Salts of
Tin "in Milligrams. . . .

RESULT

315.0
Living

Weight of Liver in Grams .

Weight of Pig in Grams . .
Weight of Liver as percen-
tage weight of Pig

Tin found in Liver in Mil-

licTTflJIlS

9.8

rmined
8 later)

220

9.46

Rate of accumulation oj
tin in liver per day in
Milliirrams

Percentage total amount
of tin found in liver. . .

34 Original Communications: Eighth International [vol.

GUINEA PIGS NUMBERS SIX AND SEVEN

These were fed no more "Salts of Tin" after 25 days, but at the end of 23
days more they died one night from exposure to cold from an open window.

Guinea Pig
No. 6

Guinea Pig
No. 7

Estimated milligrams of tin in liver at end of 25
days, calculated from average rate of accumu-
lation of tin in livers of other five Guinea Pigs . .

Milligrams of tin found in livers after 23 days rest
from feeding "Salts of Tin"

2.83
.30
.1099

2.83
.50

Average rate of elimination of tin per day in mil-
herams

.1015

No physiological examination was attempted, so that the con-
clusions from this work are somewhat limited. The following
deductions, however, would appear to be patent :

First. — That daily doses of 12.6 milligrams of tin as Hydrates
were fatal to the Guinea Pigs in from nine to twenty-five days
upwards.

Second. — That tin was progressively accmnulated in the livers
of these Guinea Pigs when fed daily doses of 12.6 milligrams of
tin as Hydrates.

Third. — That the tin accumulated in the livers of the Guinea
Pigs by feeding, as above noted, was eliminated rather slowly
after feeding was discontinued.

It is desired to give credit to W. S. Sellars who performed the
major part of the Laboratory work in connection with this exper-
iment.

SPECIAL ADAPTATION OF IODINE TITRATION

METHODS FOR THE ESTIMATION OF TIN,

ESPECIALLY IN CONNECTION WITH

DETERMINATIONS OF "SALTS OF

TIN" IN CANNED FOODS

By H. a. Bakek
■New York City, N. Y.

This method being an adaptation of parts of several standard
and well known methods, no particular reference will be made
to the sources of the principles employed in the method.

The tin in the canned food products is obtained as a sulphide
precipitate from wet combustion, with Nitric and Sulphuric
Acids, of 100 grams food product.

The clear Sulphuric Acid residue is diluted, neutralized with
ammonia, and then rendered about 2% acid with Hydrochloric
Acid, after which it is thoroughly saturated with Hydrogen
Sulphide Gas. This precipitate is then filtered on a Gooch
Crucible with a false bottom. The precipitate may contain for-
eign substances, such as Lime, Phosphorus and Silica, some
Lead, or even small amounts of Iron, but none of these will
cause any trouble subsequently in the titration so that the
labor of separating the tin completely from the precipitate is
obviated.

After washing the precipitate three or four times in a Gooch
Crucible, it is transferred to a small porcelain dish by simply
forcing out the false bottom of the Gooch Crucible and its
asbestos pad and rinsing off the crucible.

The precipitate, mixed with asbestos, is now transferred to a
300 cc. Erlenmeyer Flask and boiled with strong Hydrochloric
Acid; Potassium Chlorate being added from time to time to
insure the complete breaking up and solution of the tin sulphide,
as well as the elimination of the sulphur. This is accomplished
in a very few minutes. A few strips of pure aluminum foil, free

36 Original Communications: Eighth International [vol.

from tin, are then added to the flask until all of the Chlorine is
eliminated. This flask is then attached to a large Kipp Appara-
tus, charged with pure marble and Hydrochloric Acid, delivering
Carbon Dioxide. The Carbon Dioxide from the Kipp Apparatus ,
is passed through a Scrubber, then led into the Erlenmeyer Flask
through a bulbed tube in the rubber stopper of the Flask, de-
livering the Carbon Dioxide near the surface of the liquid in
the flask. It is led out of the flask through a second bulbed
tube, the opening of which is near the top of the flask, and the
Carbon Dioxide gas escapes from the end of a glass tube about
10" long, immersed in water about 8" deep. This gives a water
seal to the dehvery tube and a pressure against which the Kipp
Apparatus must work. This obviates any violent flow of the
gas when not desired and permits a gas pressure in the Erlen-
meyer Flask.

Pure seamless black rubber tubing and f" glass are used to
form the connections specified.

When the flask is thus attached to the Carbon Dioxide In-
sulating Apparatus, as above mentioned, the air from the flask
and the tubing connections is first thoroughly dispelled by
lifting the delivery tubes out of the water cylinder seal so that
the Kipp Apparatus has practically no pressure to overcome.
A large amount of Carbon Dioxide is thus forced through the
system and air is completely distilled. The rubber stopper in
the Erlenmeyer Flask is then raised, and about one gram of
Aluminum Foil is dropped into the flask. This quickly reduces
the tin to the metalHc form and evolves a great deal of Hydrogen
Gas.

The flask is then placed on a hot plate and heated to boiling.
The aluminum disappears and the tin is changed into Stannous
Chloride. After a few minutes boiling, the flask is set off the
hot plate and then cooled in ice water, while still under Carbon
Dioxide insulation.

When the flask is first attached to the Kipp Apparatus, enough
water is added to dilute the Hydrochloric Acid so that its strength
is approximately 30 to 40%. After the addition of aluminum
foil and boihng, the acid strength will be approximately 25 to
30%.

xviii] Congress of Applied Chemistry 37

After cooling, as above mentioned, the contents of the flask
are ready for Iodine titration. This may be accomplished by
two methods; an excess of Iodine may be run into the Erlen-
meyer Flask directly by simply lifting out the rubber stopper
and running in the Iodine solution while Carbon Dioxide is
issuing from the flask. The excess of Iodine must then be
titrated back with Sodium Thiosulphate. Usually, however, it
is satisfactory to simply detach the flask from its rubber tubes,
wash down the tubing, rubber stopper and sides of the flask
with some air free water, add starch paste and titrate directly
and quickly with N-lOO Iodine solution until a faint blue color
is obtained. The asbestos which is in the flask will not interfere
with this titration.

The air free water is made by boiUng distilled water, adding
a small amount of Sodium Bicarbonate and then a slight excess
of Hydrochloric Acid.

Only one sample has been mentioned so far, but duplicates
are always run together, as the Kipp Apparatus is arranged to
handle two flasks at a time, simply by dividing the Carbon
Dioxide Gas by means of a "U" tube and connecting an arm to
each Erlenmeyer Flask. It is not only desirable but necessary
with this method, as with practically all titration methods, to
run duplicate samples. The N-lOO Iodine is standardized against
pure tin solutions or food mixtures, such as Apple Butter, con-
taining a known or added amount of salts of tin.

The advantages which may be found iji this method are :

First: — Only one filtration is required and that can be per-
formed very quickly on account of the fact that it is performed
under suction.

Second: — No perfect separation of the tin from other metallic
precipitates or impurities is necessary.

Third: — There is no delay at any point in the method, such
as long washings, waiting for filter paper to dry, or the loss of
time over very slow and careful burning of the precipitate.

Fourth: — The titration reading gives the amount of tin
directly, as no corrections are involved.

Fifth: — In the hands of competent operators, many more

38 Original Communications: Eighth International [vol.

analyses can be performed in a given time than with a Gravi-
metric method.

Sixth: — The accuracy of the method is very satisfactory, being
at least as accurate as any Gravimetric method.

Seventh: — This same method may be used directly for metals
containing tin without previous separation. Metals, such as tin
plate, solders. Babbitt Metals or Composition Metals may be
dissolved up directly with Hydrochloric Acid in an Erlenmeyer
Flask attached to the Kipp Apparatus and titrated precisely as
above described, by either direct titration method or the excess
method, using, of course, a strong Iodine solution. Aluminum,
Zinc, Iron, Lead, Antimony, Bismuth or small quantities of
Copper do not interfere with the method. When large quantities
of metals are dissolved directly in the Erlenmeyer Flask, the
addition of Aluminum foil is not necessary. If metals are dis-
solved up in contact with the air, or Potassium Chlorate is
necessary for complete solution, they may be reduced with
aluminum foil and then handled exactly as previously described.

"SPRINGERS" IN CANNED FOODS— CAUSES AND
PREVENTION

By H. a. Baker
New York, N. Y.

"Springers" is a trade term given to cans with bulging ends
which contain perfectly sound and sterile food products. They
are undesirable because the easiest test for the Housewife to
apply, to tell whether the container is sound, is to observe that
the ends of the cans are flat or drawn in slightly. It is desirable
that this test should always be appUcable and sufficient. There-
fore, canned foods should be so packed that no "Springers" will
be formed.

When a can is a "Springer" there is too much gas in it or not
enough space to hold the gas under negative pressure at all
weather temperatures.

The gases in the head space of these "Springers" are never
more than three; Carbon Dioxide, Nitrogen and Hydrogen.
Very often no Hydrogen is found. Oxygen is practically never
found.

The Carbon Dioxide is formed in practically all canned foods
during the time of processing. It is also formed excessively if
food products are not worked through quickly from the beginning
of their preparation to their final sterilization. This is true
particularly of fruit and vegetable products after they have been
peeled or their cells have been broken in any way or have been
subjected to heat.

If food products are allowed to stand in containers before
sterilization, an excessive amount of Carbon Dioxide Gas is
formed.

Nitrogen Gas is simply a residue from unremoved air. Hydro-
gen Gas, when formed, is the product of attack of fruit or vege-
table acids on the metallic container.

Changes in temperature of these cans produce changes in gas

40 Original Communications: Eighth International [vol.

pressure. At 85° Fah. we may have a well puffed can, at 60°
one in which there is practically no pressure and at 45° to 50°
there will be a vacuum. These changes occur with a decrease of
temperature because the gas itself contracts, the solid and liquid
contents of the can contract, and the solubility of the gas is
increased.

"Springers" are usually warm weather phenomena.

The general history of the formation of gas and its behavior in
a can are as follows:

First :— A certain amoimt of air is- left in the can at the time of
sealing, even if the cans have been "exhausted."

Second: — A certain amount of Carbon Dioxide is formed dur-
ing the processing or cooking of the food products. If they have
been allowed to stand after being prepared, an excessive amount
is formed. The Carbon Dioxide is usually 8 to 15% of the gas
in the head space at ordinary temper atiu-es. At higher tem-
peratures, more of the gas comes out of solution and can be
found in the head space.

Third : — The oxygen left in the can disappears, either by com-
bination with some element of the food product, such as Butter
Fat in Milk, etc., or by combinaton with metal, or 'by com-
bination with Hydrogen Gas formed by the action of Organic
Acids on metal.

All three of these reactions have been traced. This withdrawal
of Oxygen in the can tends to give the can a temporary vacuimi.

Fourth: — When organic acids are present, Hydrogen Gas is
formed in plain tin cans and helps to make pressure.

Fifth : — The influence of the increase of heat on the expansion
of the soUd, liquid and gaseous contents of the can, and the
decrease of solubility of the Carbon Dioxide Gas are usually
responsible for the appearance of "Springers."

Sometimes Hydrogen Gas Springers are formed which are
usually old samples and could not be classed as temperature
springers.

If very small head space is left, it requires but a slight expan-
sion of the contents to change the contoiu- of the can from a flat
to a bulging condition.

xviii] Congress of Applied Chemistry 41

These difficulties can be obviated if the following points are
observed:

First : — Sufficient space must be left in the can to receive the
gases which will be formed. This means evenly filled cans in
which the exact amount of head space has been determined for
each food article and process.

Second: — This head space must be "exhausted" adequately
so that enough vacuum is left to receive the gases that will be
formed and still leave the ends of the containers drawn in or under
vacuum.

Third: — Cans, after sealing, must be processed as soon as
possible to minimize the formation of Carbon Dioxide Gas, and
there should be no undue delay in working the food product
through the factory from the beginning of its preparation until
it is sterilized.

Fourth: — With highly acid food products, the metaUic con-
tainer should have a protective coating of enamel.

APPARATUS FOR QUANTITATIVE EXTRACTION OF
THE GASES IN CANNED FOOD CONTAINERS

By H. a. Bakek

New York City, N. Y.

Figure # 1 shows the apparatus ready for use.

Figure # 2 shows the apparatus actually in use, with the
gas being collected in a regular gas burette.

The apparatus consists of an extensible strap iron frame in
which a can may be set and clamped down by means of a screw
clamp.

Entering at the base of this steel frame is a hollow steel needle
which is observable in figure # 1. The rubber stopper shown
alongside of this needle, in use, is placed over the needle. It is of
such height that the rubber stopper must be considerably com-
pressed before the steel needle punctures the bottom of the can.
The steel needle is connected with a water supply in the cylinder
having air pressure in its top. A stream of water, imder about
fifteen poimds pressure, can thus be forced through the punc-
turing needle into the can. Adequate water pressure from any
other source would be satisfactory.

The strap iron holder, which is either screwed or clamped on to
a table, has a twisted iron shank so that it tips at an angle of
approximately 45°. This places the can in such a position that it
can be punctured and the gas drawn off from its highest point
as is shown in figure # 2.

In this figure, a regular Doremus Gas Extracting Apparatus
may be seen.

Any other kind of puncturing arrangement, based on the same
principle, would answer satisfactorily.

This clamp has a screw compression which works on the
bottom of the can, and a hollow steel puncturing needle through
which the gas is extracted from the top of the can. The punc-
turing needle is enclosed in a rubber stopper so that compression

44 Original Communications: Eighth International [vol.

and tight connection is necessary before the steel needle punctures
the top of the can.

The top of the gas extractor is connected with a regular gas
burette by means of capillary tubing with rubber connections.

The complete procedure for extracting the gas is as follows:
1st — After the can is in place, water is forced through the steel
puncturing needle in the base, and while the water is still flowing
through, the screw clamp from the top is tiu-ned down imtil the
compression in the rubber stopper is great enough to allow the
needle to puncture the base of the can. Water under pressure
may then enter the can. The Doremus clamp is screwed down
solidly, the hollow steel needle filled with water, the capillary
connecting tubes between the gas burette and the extractor are
filled with waller and connected, as shown in figure # 2. The
clamps on the rubber connections are then loosened, the gas
burette opened and the Doremus clamp screwed down until the
can is punctured. The gas then flows out through the capillary
tubing, displacing the water in the gas burette.

When all of the gas has been removed, some of the liquid in the
can will come over and sweep any gas in the capillary connections
into the gas burette so thaj; quick and complete extraction of the
gas from the head space of the sample is obtained.

THE DISAPPEARANCE OF OXYGEN IN CANNED FOOD

CONTAINERS

H. A. Baker
American Can Company, New York, N. Y.

There is always left a head space in the top of every can in
which food is packed. In this head space or chamber there is
always more or less air, even when an "exhaust" has been used to
withdraw the air. The analyses of probably 100 samples of gas
from these head spaces from sound cans containing nearly all
kinds of food products have practically never shown any oxygen
content. Slight traces have been found in cans containing foods
of less than two months age.

The analyses usually show Carbon Dioxide 8 to 15% with the
balance Nitrogen Gas. Hydrogen is also found to be present in
some instances, particularly with acid fruits.

It might be presumed, in many instances, that the Oxygen
combined directly with the food product in such oily foods as
Salmon, Evaporated Milk, Pork and Beans, etc., but the
absence of Oxygen in all classes of canned foods calls for another
explanation, and in order to trace the disappearance of Oxygen
and the appearance of Hydrogen Gas, sample cans were prepared
containing distilled water, dilute solutions of Citric Acid and Salt
and their gas changes were followed by analyses.

The analyses of many samples of gas, drawn from cans contain-
ing food products, have shown the presence of Hydrogen Gas,
particularly where acid fruits were concerned. This gas does
not usually appear until after the foods have been packed two or
three months, and inasmuch as some corrosion has necessarily
been taking . place during this time, the question arises why
Hydrogen Gas does not appear more quickly.

The following experimental pack and analyses were made in
an attempt to find out the facts.

Ten regular No. 2 cans were filled as follows:

46 Original Communications: Eighth International [vol.

Two cans | full with 500 cc plain distilled water. .

Two cans | full with 500 cc solution containing 1% Citric Acid.

Two cans f full with 500 cc solution containing |% Citric Acid
and 1|% Sodium Chloride.

Two cans § full with 300 cc solution containing i% Citric Acid.

Two cans § full with 300 cc solution containing i% Citric Acid
and 1|% Sodium Chloride.

These cans were sealed cold, leaving head spaces of approxi-
mately 100 cc and 300 cc of air at room pressure and temperature
in the cans filled with 500 cc and 300 cc hquid respectively.
They were cooked one hour at a temperature of 245° Fah. and then
allowed to cool in the air. This was done on October 9th, 1911.

Analyses of the gas from five of the cans were made 42 hours
afterwards; one analysis 21 days afterwards and four analyses
8J months afterwards.

The following are the gas analyses obtained:

1. Can f full containing 500 cc plain distilled water, sealed
cold, and processed one hour at 245° Fah. — after standing 42
hours had a gas content of the following analysis :

Carbon Dioxide Trace

Oxygen 13.20%

Hydrogen .00%

Nitrogen 86.80%

2. Duphcate of this sample, after standing 8§ months, gave
the following analysis:

Carbon Dioxide Trace

Oxygen .00%

Hydrogen . 00%

Nitrogen 100.00%

3. Can I full containing 500 cc of |% Citric Acid solution,
sealed cold, processed one hour at 245° Fah. — after standing 42
hours had a gas content analyzing as follows:

Carbon Dioxide -70%

Oxygen 9.65%

Hydrogen .00%

Nitrogen 89 . 65%

xviii] Congress of Applied Chemistry 47

4. Duplicate of this sample, after standing 21 days, gave the
following gas analysis:

Carbon Dioxide . 85%

Oxygen .40%

Hydrogen .00%

Nitrogen 98.75%

5. Can f full with 500 cc solution containing }% Citric Acid and
H% Sodium Chloride, sealed cold, processed one hour at 245°
Fah. — after standing 42 hours had a gas content of the following
analysis :

Carbon Dioxide . 65%

Oxygen 9.05%

Hydrogen .00%

Nitrogen 90.30%

6. Duplicate of this sample, after standing 8§ months, had the
following gas content:

Carbon Dioxide 1 . 20%

Oxygen .00%

Hydrogen .60%

Nitrogen 98.20%

7. Can I full, containing 300 cc | % Citric Acid solution, sealed
cold, processed one hour at 245° Fah.— after standing 42 hours
had a gas content analyzing:

Carbon Dioxide -40%

Oxygen 11.60%

Hydrogen .00%

Nitrogen 88.00%

8. Duplicate of this sample, after standing 8i months, had
a gas content analyzing:

Carbon Dioxide .20%

Oxygen .00%

Hydrogen .30%

Nitrogen 99.50%

9. Can J full, with 300 cc solution, J % Citric Acid and 1^%
Sodium Chloride, sealed cold, processed one hour at 245°
Fah,— after standing 42 hours, had the following gas content:

48 Original Communications:

Eighth International [vol.

Carbon Dioxide

.40%

Oxygen

12.00%

Hydrogen

.00%

Nitrogen

87.60%

10. Duplicate of this sample, after standing 8| months, gave

the following gas analysis:

Carbon Dioxide

.30%

Oxygen

.00%

Hydrogen

.80%

Nitrogen

98.90%

The following points in connection with these analyses are
significant:

1st. Oxygen disappeared in all cans in the course of time,
although the amount originally left in the cans was very excessive.

2nd. Oxygen disappeared in cans containing plain water, also
in cans containing acid and acid and salt solutions.

3rd. Hydrogen was not found in any gas analysis until all of
the Oxygen had disappeared, although steady acid corrosion had
been going on.

If a stronger solution of Citric Acid had been used, much more
Hydrogen would have been formed and probably the Oxygen
would have disappeared more quickly.

The following analyses were obtained on commercial samples
of food products :

Can, eighteen months old, containing Red Raspberries, furn-
ished gas of the following analysis:

Carbon Dioxide 8.40%

Oxygen .00%

Hydrogen 65.50%

Nitrogen 26 . 10%

Can, nine months old, containing Red Raspberries, furnished
gas of the following analysis:

Carbon Dioxide 10.90%

Oxygen .00%

Hydrogen 16.50%

Nitrogen 72.60%

xviii] Congress of Applied Chemistry 49

Can, eighteen months old, containing Strawberries, furnished

gas of the following analysis:

Carbon Dioxide

12.60%

Oxygen

.00%

Hydrogen

72.40%

Nitrogen

15.00%

Can, one year old, containing

Strawberries,

the following analysis:

Carbon Dioxide

13.20%,

Oxygen

.00%

Hydrogen

27.20%,

Nitrogen

69.60%

These containers were not properly protected by means of
enamel, so that corrosion had been very excessive.

No analyses of gases from canned food containers have ever
shown Hydrogen and Oxygen gas together, and inasmuch as
Hydrogen Gas must necessarily be formed continuously from the
beginning, when acid fruits are present, it would appear that the
Hydrogen, under the conditions obtaining in a tin container,
combines with Oxygen, and consequently cannot be found until
all of the Oxygen has disappeared.

It would appear, therefore, that Oxygen disappears in tin
food containers in at least the three following manners:

1st. By combining with the metals tin and iron, forming oxides.

2nd. By oxidizing tin or iron salts.

3rd. By combination with Nascent Hydrogen, when organic
acids act on the metallic container.

It is also probable, in some instances, that Oxygen combines
directly with the food product, during processing, particularly
with such foods as Evaporated Milk, canned Salmon, Pork and
Beans, etc. in which there are oily substances. In evaporated
Milk and Pork and Beans, there is some caramelization which
would also take up some Oxygen.

The analytical work reported in this paper was done by W. S.
Sellars.

EGGS P]

For several
used for pres(
Stations,* the
scientists. W
to learn, but
Thieriot in 1!
and obtaining
This method ]

52 Original Communicqitions: Eighth International [vol.

part NajO to four parts Si02. The silicates containing such
large proportions of Na20 as the first mentioned would be too
strongly alkaline for keeping eggs.* It has been found that eggs
kept in a strongly alkaline solution absorbed some of the alkali
and produce a jelly like condition of the whites. It is probable
that the Si02 is not readily deposited from such solutions and
the pores of the shell are not closed immediately, consequently
some of the solution finds its way through to the interior and the
property to which water glass owes its efficiency as a preservative
is lost. The writer has obtained very satisfactory results with a
siUcate containing 24 . 2 parts of Si02 to 8 . 89 parts NagO made

Six eggs weighed

when put in boI-

ution

Six eggs weighed

when taken out of

solution

No. 1

67.18 gms.
55.85 "
62.40 "

55.76 "
56.67 "

53.77 "

57 . 12 gms.
55 85 "

No. 2

No. 3

62 35 "

No. 4

55.75 "

No. 5

56 62 "

No. 6

53 75 "

341.63 "

341.44 "

into the glass by the dry process then dissolved by superheated
steam and made up to a syrup testing 38 degrees B. One part
of this syrup to nine parts of water makes a solution of about
0 . 045 specific gravity, in which fresh eggs readily sink and will
remain submerged. Eggs kept in this way are of better flavor
than cold storage stock. They never have the musty taste so
often found in the storage goods and about the only difference
between them and fresh eggs is a little lack of flavor. The
shells are hermetically sealed and no bacteria can get through
them, neither can oxygen, consequently if they contain any fife
when put in the solution it is very soon destroyed. Six fertile
eggs put in a jar of water glass were kept in an incubator for
6 days at a temperature of 103 degrees F. and an examination

* Bomtraeger (Oeslem. Chem, Ztg. 3, 1900, No. 12, p. 295.

xviii] Congress of Applied Chemistry 53

at the end of that time was made, showing that the embryo
had made no growth. There is very little change in the moisture
content, and, unlike eggs in cold storage, the weight remains
practically constant.

The writing of this paper and the limited investigation which
is here given was suggested by a newspaper article which con-
tained the statement that eggs preserved in water glass were
unfit for food because they contained quite a quantity of soluble
silica which if taken into the system was very dangerous and
liable to cause coagulation of the blood. Notwithstanding the
ridiculousness of the statement many people were alarmed and
ceased to use eggs preserved in the siUcate, and often inquiries
were made to learn if any investigations had been made to deter-
mine if silica passed into the egg content. The work, therefore,
was undertaken to determine principally whether eggs kept in
water glass contained any more siUca than fresh eggs. Some
work was also done to determine if any marked changes take
place in the nitrogen compounds. The most noticeable physical
change in the eggs is a thinning out of the white which after
the egg is kept 10 or 12 months does not coagulate so firmly
as does that of a fresh egg, and the white appears much more
watery.

The results of the investigation are given in the tables which
follow. To separate the yolks from the whites completely, par-
ticularly in the preserved eggs, it was found necessary to boil
the eggs before breaking them which, of course, caused some
loss of moisture. The methods of analysis used in the experi-
ments were those employed by the Bureau of Chemistry, U. S.
Department of Agriculture,* in the work on cold storage eggs.
The preserved eggs used weie put down by the writer in a 10
per cent solution of 38 degrees B. water glass syrup, containing
one part NajO to 2.7 parts SiOj, in April 1911. They were,
consequently, when examined about 11 months old. The other
lot was put in the same kind of a silicate solution in February
1912 and examined in April, consequently were about two
months in the solution.

* Bulletin No. 115, Bureau of Chemistry, U. S. Dept. of Agric.

54 Original Communications: Eighth International [vol.

Weight op Eggs, Whites and Yolks and Loss in Boiling

-otM

•s

•3

III'

Grams

Grama

Grams

Per cent

3 fresh eggs

171.272

166.838

17.93

88.51

59.301

2.59

3.81

3 w, g. eggs, one

193.353
175.102

190.199
170.304

22.835
18.071

97.86
98.683

63.115
52.738

1.63
2.74

4.93

3 fresh eggs

3.20

3 w. g. eggs, 2

months

175.320

169.550

16.91

96.81

51.92

3.29

5.52

Partial Analysis of Fresh and Preserved Eggs
Wet Basis

Nitrogen present as

3 a

11
11

t§

Per cent

Percent

Per cent

Percent

Fresh Eggs

White

87.40

0.785

0.005

1.78

1.585

0.195

0.115

0.08

Yolk

48.05

1.35

0.062

32.15

2.655

2.540

0.115

0.015

0.10

Preserved

eggs. 11

months. .

White ....

85.15

0.70

0.006

0.02

2.05

1.675

0.375

0.365

0.11

Yolk

52.80

1.30

0.060

29.70

2.37

2,21

0.160

0.008

0.152

2 months

in water

glass. .. .

White ....

86.43

0.73

0.012

0.054

2.018

1.783

0.235

0.172

0.063

Yolk

50.25 1.47

0.040

30.68

2.53

2.405

0.125

0.015

0.110

xvin]

Congress of Applied Chemistry

Partial Analysis of Fresh and Preserved Eggs
Dry Basis

Nitrogen present as

33
6

Fresh Eggs.

White

Yolk

Preserved eggs in
water glass. 11
months.

White

Yolk

Preserved eggs in
water glass
months

White

Yolk

Per cent

6.23
2.64

4.71
2.46

5.37
2.93

Per cent

0.04
0.119

0.04
0.113

0.087
0.080

Per cent

62.98

0.134
56.25

0.398
61.04

Percent

14.12
5.20

13.80
4.49

14.88
5.04

Percent

12.68
4.

11.29
4.19

13.14
4.81

Percent

1.54
0.225

2.58
0.30

1.732
0.23

Percent

0.912
0.029

2.46
0.15

1.267
0.03

Percent

0.64
0.195

0.74
0.288

0.464
0.22

In addition to the results given in the lables some work was
done in coagulating the albumen of the white with different
reagents. Three eggs of each kind were taken and the whites
separated as completely as possible from the yolks in the raw
condition. 10 gram samples of the whites were treated with
acidulated water and boiled. The whites yielded copious floc-
culent precipitates which were thrown on tared filters, washed
with hot water, dried and weighed.

The fresh eggs yielded dried albumen 10 . 0 per cent.

The 11 months old preserved eggs yielded dried albumen
10 . 28 per cent.

Samples treated with alcohol and allowed to stand several

hours yielded :

Fresh eggs, dried albumen 12.00 per cent.

Preserved eggs, dried albumen 12. 39 per cent.

66 Original Communications: Eighth International [vol.

Calculated from nitrogen content (n. x 6 . 25)

Fresh eggs = 11 . 37 per cent.

Preserved eggs = 11 .62 per cent.

It is obvious from the figures here obtained and those given
in the table that there is practically no difference in the total
coaguable proteid matter of the fresh or preserved eggs. There
seems to be a slight difference in the amount of nitrogen or pro-
tein coagulated by heat alone and is probably due to the pres-
ence of albumoses and peptones which are absent in fresh eggs
but appear to develop as the esgg age.* This same change was
noted in the case of cold storage eggs, and reported in a paper
published by Dr. H. W. Wiley and others, t

Conclusions

1. Eggs packed in the right kind of water glass (silicate of
soda) contain no more siUca or other ash materials than fresh

2. The moisture content remains constant and a preserved
egg weighs practically the same as when put in the solution.

3. The nutritive value as far as one can judge from the
chemical analysis is the same as that of a fresh egg. The quahty
is superior to most cold storage eggs, as the pores of the shell
are closed and no bad odors or flavors are absorbed.

' Allen's Commercial Organic Analyses, Vol. IV, p. 41.

t Bulletin 115, Bureau of Chemistry, U. S. Dept. of Agric., p. 32.

SOME OF THE RESULTS OF THE FOOD AND DRUGS

ACT

W. D. BiGELOW, Ph.D.

Chief of Division of Foods and Assistant Chief, Bureau of Chemistry,
U. S. Department of Agriculture, Washington, D. C.

The complete study of this question would involve the principles
governing the manufacture and sale of each particular kind of
food and drugs. It would necessitate the discussion of the
history and development of a large number of articles and of the
varied influences which lead to the adoption and subsequent
discontinuance of individual practices, in each. This would lead
us into a maze of details which would require a considerable vol-
ume for their adequate presentation. In a paper of this scope,
therefore, it is only possible to consider some of the general prin-
ciples involved and a few of the fundamental changes that have
resulted from the enforcement of the Act of June 30, 1906.
At the time of the passage of the Food and Drugs Act, notwith-
standing the creditable work that had been done by 27 States in
the enforcement of their laws, labels on foods were very frequently
so written as to deceive the consumer with respect to the charac-
ter, value or origin of the product.

When the labels purported to give the weight of the product
their statement was commonly exaggerated, sometimes being
the gross weight of the product and package and sometimes
having no relation to the weight of the product at all. Cans
known technically as No. 1, 2, 3, etc., were sometimes desig-
nated as "1 pound," "2 pound," "3 pound," etc., though holding
perhaps only two-thirds of that amount. Bottles of wine, oil
and other products measuring five to a gallon, — sometimes a
smaller amount, — were often labeled as quarts. Canned food
and bottled goods are still sometimes referred to in grocers'
lists and restaurants as "Pounds" and "Quarts," respectively,
but the practice of designating them in that manner on the label
and on the shipping case has been discontinued.

58 Original Communications: Eighth International [vol.

There are many difficulties encountered in marking the weight
accurately upon a package of some kinds of food and it is prob-
able that the misstatements that now exist on labels of food with
respect to weight are often unintentional. The fraudulent state-
ments which were prevalent five years ago have almost dis-
appeared.

Closely related to misbranding with respect to weight is the
practice in packing of canned food usually known as "slack
filling." In such case the can was only partially filled with the
food in question, the deficiency usually being made up with water
to prevent the collapsing of the can. For instance, peas or beans
were filled to within one-half inch, or possibly sometimes an inch,
of the point to which they should be filled and then water or weak
brine added to make up the proper volume. In packing cove
oysters — in the extreme illustration of slack filling — only one
and one-half ounces of oysters were sometimes placed in cans
capable of holding over five ounces and the cans were then filled
with brine. In tomatoes the cans were filled probably to within
an inch of the top or sometimes only half or two-thirds full and the
deficiency made up with water or perhaps weak brine. Steps
were taken to correct this abuse and during the last year success-
ful prosecutions have been maintained against packers of slack
filled cans. The Department has announced publicly that this
practice is fraudulent and it is believed that it has now been
entirely discontinued.

Foods have been misbranded commonly with respect to the
name of the place, (i. e. the country or region) in which they were
produced or manufactured. This practice has obtained for two
reasons: first, because of the desire of the packer in one locality
to take advantage of a favorable reputation of another locality
and so misbrand his goods as to the place of their manufacture;
second, because of a certain glamor which a foreign name pos-
sesses for many consumers. As an illustration of the first class
of abuses with respect to geographical name may be mentioned
the packing of "Maine Sweet Corn" in Maryland, of" Michigan
Apples" in Arkansas, of "California Canned Fruit" near the
Atlantic Coast, of "Minnesota Flour-" in the mills of Iowa and
Missouri.

xviii] Congress of Applied Chemistry 59

This form of misbranding gradually shades into the class where
the misuse of a geographical name causes a false impression with
respect to the material of which the food is made; for instance,
the term "Vermont Sirup" or "Ohio Syrup" means maple sirup
to the consumer because that is the only sirup made in Vermont
and Ohio. These terms have been used frequently on the label
of a cane sugar sirup colored and sometimes flavored in imitation
of maple sirup. This form of misbranding has been corrected
generally with reference to the staple articles of the United
States.

Material progress has been made in correcting this form of
misbranding in the case of foreign products and of foods manu-
factured in the United States in imitation of foreign products.
Some of the brands of coffee which were formerly labeled "Mocha
and Java," for instance, are now merely called "Coffee." Im-
porters have been required to discontinue, on imported foods,
the language of another country than that of their manufacture.
Progress has been made in the correction of the label of imported
wines which are commonly misbranded with respect to their
character or class. Progress has also been made in the correction
of the labels of certain products manufactured in the United
States in imitation of foreign products, such as macaroni and
tomato paste. Several kinds of cheese manufactured in imitation
of well known foreign varieties are now labeled with the place
of their manufacture. Rice grown in this country from Japan
seed is labeled as grown in the United States. A product formerly
called "Holland Rusk" with the label embellished with Dutch
scenery is now labeled as made in Holland, Michigan, and the
Dutch windmill has been removed from the label. This form of
misbranding again merges into adulteration as, for instance, when
cottonseed oil grown in the United States is placed in decorated
tins so labeled as to represent the product to be an Italian olive
oil. These practices still obtain to a certain extent, though to a
much less degree than formerly.

One of the prevalent forms of misbranding is the use on the
label of exaggerated claims regarding the strength and the nutri-
tive value of the product. This form of misbranding is especially
applicable to proprietary remedies but has also been practiced

60 Original Communications: Eighth International [vol.

largely with foods. Flavoring extracts were often labeled "Double
Strength," or "Triple Strength," although the products so labeled
were rarely beyond standard strength, and not infrequently
were entirely fictitious. Breakfast foods and infant foods carried
on their labels a glowing description of their miraculous nutritive
value and sometimes curative properties. Cereal preparations
of ordinary composition without any of the starch having been
removed were sold under labels representing them to be diabetic
foods. These practices have largely passed away as far as the
labels are concerned. Unfortunately, the law does not reach
posters and advertising matter sent through the mails and by
such means fraudulent statements regarding the quality and
nutritive value of some preparations is conveyed to the consumer.
The labels themselves, however, have been greatly improved.

The addition to foods of substances of lower value to serve as
a make weight and thus cheapen the food has largely been dis-
continued. At the time of the passage of the Food and Drugs
Act such practices had been made away with in a number of the
States but in other States and indeed in interstate commerce
they were still quite prevalent. As illustrations of this practice
may be cited rye flour and buckwheat flour, both of which con-
tained a substantial amount of wheat flour; spices which were
commonly loaded with cereal preparations, ground olive stones,
cocoanut shells, etc., a line of preparations being manufactured
and sold for the purpose of adulterating spices and pepper shells
and olive stones being imported into this country for that purpose.

At the time of the passage of the Food and Drugs Act there was
little pure maple syrup manufactured commercially and sold in
interstate commerce. Immediately after the law went into effect
there was practically no maple syrup to be had but brands of
so-called maple syrup which had formerly borne on the label
an offer of a large reward to anyone who would prove the presence
of any adulterant appeared under a new label, declaring the
contents of the package to be a mixture of maple syrup and cane
sugar syrup.

Cider vinegar was commonly diluted with water to reduce
its strength in acetic acid to the desired percentage and since this
dilution brought the solids content down to a lower figure than

xviii] Congress of Applied Chemistry 61

that in commonly accepted standards a quantity of boiled cider
was added. It only remained to add a larger quantity of boiled
cider and, of water and then to strengthen with distilled vinegar
to obtain much larger yields and this practice merged gradually
into the preparation of an entirely fictitious product manufac-
tured from distilled vinegar, with color added and solids in the
form of boiled cider. The detection of practices of this kind by
analytical means offered many difiicult problems which have
been partly solved and maple sirup and cider vinegar may be
cited as types of a large number of products which are now sold
in the pure state to a very much larger degree than was true at
the time of the passage of the Food and Drugs Act.

The two classes of substances relied on chiefly by the manu-
facturer in the preparation of fictitious products are colors and
flavors. It was a difficult matter to handle either of these classes
of substances in such a way as to imitate a natural food. The
manufacturer who uses them is likely to go to extremes and the
fictitious products he puts on the market are frequently of a hue
that is scarcely to te found in nature, whereas the flavors are also
commonly in excess. The improvement in natural products
that has attended the work of the last five years, accompanied
by the better information of the public regarding such matters,
has resulted in a growing aversion for fictitious colors and fiavors
and many lines of products which were formerly in demand are
now regarded by the public with disfavor. Moreover the whole-
someness of the colors employed has been considered. The
Department has authorized the use in foods of a Ust of 7 coal
tar colors and these must be manufactured in such a way as to be
free from arsenic and other deleterious substances. Manufac-
turers have to a large extent complied with this regulation.

To a much greater extent than ever before manufacturers are
giving attention to the question of the wholesomeness of sub-
stances used in the preparation of foods. Formerly this was not
the case. When it was desired to begin the use of a preparation
in the manufacture of foods the ordinary article of commerce was
frequently employed without any thought of its possible injurious
properties. When acid phosphate was employed, for instance,
in the preparation of a food or drug the acid phosphate of com-

62 Original Communications: Eighth International [vol.

merce was used and it was not known that it contained a consid-
erable amount of arsenic. Notwithstanding the fact that lead
pipes have been known for generations to be improper for con-
ducting water for household purposes, they were employed for
tartaric and citric acid which were intended to be used as foods
and a relatively large amount of lead thus found its way into the
product placed on the market for ordinary consumption.

When a confectioner desired to give a gloss to some of his
wares it occured to him that the product used by the painter
would meet his requirements and he took ordinary shellac with-
out considering whether the lac itself was injurious to health and
without thinking of the fact that the shellac of commerce contains
a considerable amount of arsenic. We even found a large shipper
of green coffee who desiring to polish his wares and give them a
faint yellowish shade, used the first yellow powder which came
to his attention and this happened to be chromate of lead.
When it was desired to prolong the life of certain foods and at
the same time make unnecessary the care in handling which
would otherwise be necessary among the preservatives suggested
and largely employed are some substances whose toxicity was
universally admitted; e.g. — formaldehyde and ammonium fluorid.

The important point is not that sopie practices of this nature
have been corrected and others are being corrected at this time,
but that there is a rapidly growing tendency on the part of manu-
facturers when considering the use of a new or unusual substance
or preparation in the manufactm-e of food to consider whether
it is injurious to health, either because of its nature or compo-
sition or because of certain impurities and whether for any reason
its addition to food is objectionable.

The removal of the manufacture of prepared foods from the
home to the factory has made great changes in our civiUzation
and made necessary precautions which were before unthought of.
One of the most prominent characteristics of civihzation is the
increased emphasis placed on cleanliness and sanitary conditions.
It is only this fact which has made possible in the home during the
last century the preparation of many of the domestic preserved
foods which are now most prevalent. The manufacture of these
foods in the factory, however, has not been confined to men who

xviii] Congress of Applied Chemistry 63

were qualified to enforce sanitary conditions such as are necessary
for the successful preparation of many articles of food. The
result has been that we have had placed on the market on the one
hand preparations in a more or less advanced state of decay and
on the other hand substances contaminated with pathogenic
organisms. Thus there have been cases of contaminated water
being bottled and sold as spring water and being used for the
preparation of soft drinks and for serving from soda fountains.

Tomato catsup has been prepared from the peeUngs and cores
of unwashed tomatoes, including a considerable part of rotting
material, and by a process and amid surroundings which caused
additional decomposition to take place during the course of
manufacture. Ripe olives and figs were often imported into the
United States in a wormy and decomposed condition. So little
attention was given to the matter that it was the custom of rail-
roads to sell unclaimed food products resulting from wrecks to
the highest bidder with a knowledge that they would be placed
on the market indiscriminately.

Badly contaminated water has been used for cleansing milk
cans in dairies and together with contaminated ice has not in-
frequently been added to the milk. The sanitary condition of
dairy stables has frequently been bad. Eggs so far advanced
in decomposition that they would not be used by a housekeeper
have been broken in large quantities and placed on the market
either dried or frozen. Oysters and clams have been taken from
contaminated water and placed on the market with the inevitable
result of spreading typhoid fever.

These sanitary problems offer difficulties which cannot be
overcome in a day but in all of them material progress has been
made. A number of the States, realizing the importance of
sanitary requirements, have enacted special sanitary laws whose
enforcement has done much to improve the conditions formerly
existing. Of still greater importance, however, is the fact that
manufacturers as a whole have become interested in the deside-
rata of a food manufacturing establishment from a sanitary
standpoint and the changes resulting in the cleanliness of their
estabhshments, as well as in their utensils and the character of
the raw material they employ, are among the most satisfactory

64 Original Communications: Eighth International [vol.

results of the recent enforcement of food legislation. Whereas
formerly only rule of thumb methods were employed we now meet
chemists, bacteriologists and microscopists in many general food
manufacturing establishments. It is frequently made the duty
of some special officer to study and be responsible for the sanitary
condition of the factory. The health of the employees is con-
sidered with reference to the influence it may have upon the food.
Cleanliness is more frequently reqxiired, as well as uniforms or
special factory clothes, and in some establishments manicurists
are employed.

The nmnber of prosecutions that have been successfully main-
tained for the violation of the law is of minor importance com-
pared with this change in the attitude of the manufacturers.

SUR L'ANALYSE DU PHOSPHORE DANS LES
CENDRES DU LAIT

M. LE De. Bokdas
College de France, Paris, France

L'exp6rience nous a d6montr6 que I'acide phosphorique exist-
ant k r^tat de phosphates dans les cendres d'un lait correspond
k la totality du Ph contenu dans ce liquide, c'est-3,-dire au Ph
mineral et au Ph organique: l^cithine, nucl^ine, etc. . . .
Cette particularity est tr6s importante k connaitre pour ^Adter
certaines erreurs d'appr^ciation sur la valeur alimentaire du lait.

II s'ensuit done, pour le cas particulier du lait de vache,
lorsqu'on fait les cendres de ce liquide on ne provoque aucune
disparition de phosphore par Taction du charbon sur les phos-
phates, et la mati^re grasse n'entraine aucune partie du phos-
phore k l'6tat de combinaison volatile. II est inutile d'ajouter
des sels de chaux, de baryte, de magn^sie, etc., comme le pr4-
conisent plusieurs auteurs, pour 4viter des pertes en phosphore
par calcination.

Le phosphore total d'un lait pent done 6tre dos6 directement
sur ses cendres. D'autre part, en precipitant le lait par I'acide
trichlorac^tique on determine le Ph mineral dans le lactoserum
et le Ph organique dans le coagulum.

L'ACIDITE ORIGINELLE DU LAIT

M. LE Dk. Bordas
College de France, Paris, France

Les auteurs ne sont pas tous d'accord sur la reaction k attribuer
au lait, pour les uns ce liquide, k V6tat frais, serait acide, pour
les autres, il serait amphotfere, c'est-^-dire possdderait une
reaction alcaline et une reaction acide.

En 6tudiant cette question nous avons constats que ces diver-
gences d'opinions r(5sidaient uniquement dans I'emploi d'indi-
cateurs qui ne r^pondaient pes aux conditions exp6rimentales.

Nous avons 6tabli que la phtal^ine du phenol est I'indicateur
de choix pour 6tudier la reaction du lait. Lorsque ce liquide est
pr6cipit6 par notre r^actif alcool 65° ac6tique k 1-1000 nous
constatons, en tenant compte de racidit6 du r&ictif, que I'acidit^
totale d'un lait frais se retrouve dans le coagulum et qu'elle est
due exclusivement k la cas^ine libre.

L'exp6rience nous a d6montr6 ^galement qu'il existe dans un
lait frais aucun acide libre, lactique, citrique, ni aucun sel k
fonction acide, que I'augmentation de l'acidit6 d'un lait pro-
vient tout d'abord de la cas^ine d6plac6e, de sa combinaison
calcique, par Taction de Tacide lactique form6 aux d^pens du
lactose et que I'acidit^ lactique n'apparait ensuite que lorsque
cet acide a r6agi sur les sels de chaux du lait.

DE L' ACTION DU LAIT SUR CERTAINS REACTIFS

M. LE Dr. Bobdas
College de France, Paris, France

Pour expliquer les ph6nom6nes p6roxydasiques obtenus au
sein d'un liquide on s'appuie en g6n6ral sur I'existence de sub-
stances diastasiques que certain consid^re comme une indi-
viduality d^finie. Or, jusqu'ici, il n'a pas 6t6 possible d'isoler
k I'^tat de puret6 les matiSres diastasiques actives et toujours
nous les retrovons k c6t6 d'616nients min^raux. II existe done
une relation 6troite entre tous ces 616ments et leur ensemble
constitue un systSme p^roxydasique que nous retrouvons dans
r^tude des diastases du lait.

Le but que nous poursuivons consiste k rechercher le m6ca-
nisme qui preside aux reactions color6es obtenues dans le lait
avec certains r^actifs.

Nous allons nous occuper, en particulier de Taction de la
paraphdnylfenediamine sur le lait, ce corps formant le r^actif le
plus sensible pour la recherche des p^roxydases du lait.

Dans cette 6tude nous avons 6t6 amends k reproduire artifi-
ciellement des ph6nom6nes p6roxydasiques k I'aide de sub-
stances prises souvent en dehors des mat^riaux existant dans
le lait. -

On sait que la paraph6nyl6nediamine par oxydation forme de
la quinone. Cette oxydation se produit d6ja en laissant exposer
k I'air une solution aqueuse de cette base qui devient plus ou
mains brune suivant le temps d' exposition. On arrive 6galement
k ce r^sultat par Taction d'un courant d'O ou par la decom-
position de TH*0* au sein d'une solution aqueuse de cette dia-
mine, mais on obtient aucune coloration bleue comme celle qui
se produit dans un lait frais additionn6 d'H^'O" et de para-
ph6nyl6nediainine. Cette coloration n'est done pas le r^sultat
d'une simple oxydation, il est en effet n^cessaire de faire inter-
venir une autre cause pour expliquer la coloration bleue.

70 Original Communications: Eighth International [vol.

Nous avons constats qu'elle 6tait due ^ Taction d'ua produit
interm^diaire entre la paraph^nyl^nediamine et la quinone sur
les sels de chaux.

II nous suffit par exemple, de verser une goutte d'une solution
de paraph^nyl^nediamine sur un b&ton de craie pour obtenir
imm^diatement une coloration bleue.

Ceci nous conduit k obtenir directement cette coloration avec
certains sels de chaux en m^me temps que ces sels nous servant
de catalyseurs de H'O^ pour I'oxydation de la paraph^nyl^ne-
diamine.

Prenons, en effet, du citrate de chaux on du phosphate tri-
calcique purs et sees, ajoutons 2 ou 3 gouttes d' H^O^ et melan-
geons, si on ajoute ensuite une goutte ou deux d'une solution
fraiche de paraph^nylSnediamine £i 2% on obtient imm^diate-
ment la coloration bleue en question qui se fixe sur le sel de
chaux. Si avant d'ajouter le r6actif on d^laye dans I'eau les
sels de chaux insolubles et oxygen^s on constate encore la colora-
tion bleue qui reste tou jours fix^e sur la chaux.

Cette reaction se divise done en deux phrases:

1° — Oxydation de la base par un ph^nomfene catalytique.

2° — Coloration bleue produite par la chaux du sel sur le
r^actif oxyde.

Nous avons toujours obtenu ces r6sultats avec des poudres
sfeches impr^gn^es d'H^O^ et renfermant de la chaux, et toutes
les fois que la chaux n'^tait plus en presence la reaction 6tait
negative.

Les experiences suivantes sont en effet tr&s concluantes.

Si dans deux petites capsules on place dans I'une de la pierre
ponce ordinaire et dans 1' autre la m^me pierre ponce trait6e par
I'eau r6gale, lav6e k I'eau et sech6e k 100°, puis qu'on r^pete
Texp6rience pr6c6dente, on constate que les grains de pierre
ponce ordinaire sont seuls color6s. Les mgmes r^sultats sont
^galement obtenus avec de la cas^ine priv6e ou non de sa chaux
comme nous I'avons signals dans un travail ant^rieur. Nous
ferons remarquer en outre, que cette coloration bleue ne se
produit que dans un milieu tr^s l^gferement acide.

En r6sum6, nous pouvons admettre, d'aprSs ces experiences,

xvin] Congress of Applied Chemistry 71

que I'oxydation de la paraph6nyl6nediainine produit vm laque
bleu-indigo en presence d'un sel de chaux.

Voyons maintenant ce qui se passe lorsqu'on recherche les
p6roxydases du lait au moyen de la paraph^nylenediamine.

D'apr^s les theories actuelles il existerait dans le lait frais-des
diastases capables de decomposer I'H'O* et I'oxygfene mis en
libert6 provoquerait la coloration bleu-indigo de la paraph6nyl-
Snediamine, d' autre part, on salt qu'un lait port6 k 80° perd la
propri6t6 de decomposer I'eau oxyg6nee.

La modification apport6e par la chaleur dans un lait frais est
due k la coagulation d'une partie de la mati^re prot6ique qui
empfeche la decomposition de I'H^O* ajoutee au lait, mais nous
sommes parvenus h, isoler neanmoins le catalyseur d'un lait
bouilli au moyen de la centrifugation comme nous I'avons
demontre ant6rieurement, c'est-^-dire, en recueillant le d^pdt
forme au fond du tube du centrifugeur et la cr^me qui surnage.
Ces deux parties du lait bouilli ou sterilise sont capables de
decomposer TH^O'' et oxyder la paraphenylfenediamine donnant
la coloration bleue en presence de la chaux.

Une experience nouvelle plus concluante encore nous a per-
mis d'obtenir une reaction positive avec la paraphenylfinedia-
mine sur le lait entier chauffe k plus de 80° mais homogeneise h
I'aide d'une machine pulverisant le lait k une pression de 200
atmosphkes. Cette operation ayant pour effet de donner k
toutes les molecules, des corps insolubles contenues dans le lait
une mfeme dimension. On retabUt ainsi, dans une certaine
mesure, I'etat colloidal primitif du lait et son catalyseur a pu
agir k nouveau sur I'H^O* et donner en presence de la para-
phenylSnediamine la coloration bleue.

Nous ecartons dans cette experience comme dans toutes
celles que nous avons faites sur les laits bouillis les causes
d'erreur dues k la presence de bacteries ou de muscedinees qui
peuvent exister lorsqu'on opere avec des laits alteres.

Nous avons constate que la reaction avec du lait homogeneise
est moins intense qu'avec du lait frais mais il n'en reste pas
moins etabli que nous pouvons redonner par un procede meca-
nique k du lait chauffe k plus de 80° son pouvoir peroxydasique.

Nous avons encore demontre avec ces laits que la coloration

72 Original Communications: Eighth International [vol.

bleue du r^actif employ^ est bien due a la presence des sels de
chaux du lait. En efEet, en prenant un lait fix6 et st6rilis6, nous
pouvons lui rendre son maximum d' action en introduisant dans
ce lait un catalyseur artificiel, soit une poudre pulv^rulente
insoluble sans action chimique comma la Sio" pure par exemple,
soit des traces d'une solution d' oxalate de fer. Dans ces condi-
tions la coloration bleue avec la paraphenylenediamine devient
trSs intense.

Toutes ces experiences d^montrent bien que les reactions
negatives avec le r^actif k la paraph^nyl&nedi amine dans im
lait chauffe k 80° ne sont dues qn'h un changement d'etat phy-
sique du lait et que les peroxydases ou les catalases qui ont 6t6
signages, et qu'aucun auteur n'a pu isoler h I'^tat de puret6
doivent 6tre consid6r6es jusqu'^ present comme des combinai-
sons organo-m^talliques jouant un role chimique et non bio-
logique.

A CHEMICAL INVESTIGATION OF ASIATIC RICE

By Allerton S. Cushman and H. C. Fxtller

Institute of Industrial Research, Washington, D. C.

Introduction.

The following paper contains a description and the results of
a complete chemical investigation of twenty-seven samples of
Asiatic rice, which was recently carried out at the instance of
the Siamese Government. The samples were collected in the
open market at Singapore and Shanghai and no effort was made
to prepare them in any way differently from those rices which
are ordinarily exposed for sale in the Asiatic market. The
relation of an exclusive rice diet on the etiology of beri-beri
disease has been much discussed for a number of years past.
This paper does not pretend to decide this controversy but is
offered as a contribution to the general knowledge of the chemical
constitution of rice. As far as the authors are aware the results
on the phosphate content of eastern rices is the most complete
as yet published.

Description of Samples.

The samples reached the Institute on October 30th, 1911,
and the box containing them was opened on October 31st. The
samples were contained in twenty-seven 10 pound cotton bags
numbered serially 1 to 27. No other distinguishing marks or
information was found.

The cotton bags were found to be frail and rotten and ia some
cases were broken through, so that the contents had partially
escaped. All the samples contained living weevils, and a few
worms and beetles were also found. The condition of the sam-
ples made it necessary to hand pick them to remove insects.
They were then immediately packed in glass bottles, stoppered
and labeled.

The appearance of the samples indicated that they represented
a medium grade of white or milled rices. On the trip from the
4 73

74 Original Communications: Eighth International [vol.

Far East the samples had evidently suffered desiccation with the
result that some of the grains had become abraded and broken.
As it was not believed, however, that the grain had suffered in
such a way as to affect the chemical analysis except in regard
to moisture content and the weight per 100 grains, it was de-
cided to be unnecessary to delay the investigation by awaiting
a new importation of samples from the Far East.

The Analytical Work.

The analytical work was carried out by the methods recom-
mended by the Association of Official Agricultural Chemists
of the United States, and comprised the following elements
usually sought: Moisture, Ash, Proteids, Ether Extract (mainly
Fat), Fibre, Starch and other Carbohydrates, Weight per 100
Grains.

The above determinations have usually been accounted
sufficient to fix the nutrition value of a given cereal. In view,
however, of a recently published claim that milled rices are
deficient in organically combined phosphorous, phosphate deter-
minations were carried out on each sample. The results have
been carefully checked and may be taken as accurate for the
samples worked on.

Tabulation of Results.

The results of the analytical work on the twenty-seven sam-
ples submitted are given in Table I, with the exception of the
phosphate contents which are tabulated separately in Table III.
Table II gives the results of analysis of two fresh samples of
South Carolina (U. S. A.) rices bought at a prominent grocery
house in Washington, D. C. These samples are denominated
Numbers 29 and 30. Sample 29 is the ordinary very white large
grained rice as sold in the United States at about ten cents a
pound. Sample 30 was sold for a sKghtly higher price and pur-
ported to be a "natural uncoated special pure rice." Table III
gives the phosphate content of all samples, reported as phos-
phoric anhydride, PaOs. In Appendix A are given the results
of an examination of various rices exhibited at the World's
Columbian Exhibition, at Chicago, in 1893, the analyses made by

XVIIl]

Congress of Applied Chemistry

the Division of Chemistry, U. S. Department of Agriculture.
Appendix A is preceded by an extract from Bulletin No. 13, and
is followed by a summing up of the results.

TABLE I

Resttlts"' of Analysis op Twenty-Seven Samples op Rice Submitted

TO THE Institute op Industrial Research by the Siamese

Legation, Washington, D. C.

Sam-

Ether

Starch

ple

No.

Weight of

Mois-

Ash

Ex-

Crude

Pro-

and Car-

100 Grains

ture

tract

Fibre

teida

bohy-

drates

1 . 565 grams

11.02%

0.46%

0.31%

0.40%

8.13%

79.68%

1.39

10.99%

0.51%

0.29%

0.60%

8.25%

79.36%

1.181 "

11.11%

0.56%

0.20%

0.29%

7.38%

80.46%

1.036 "

10.82%

0.46%

0.15%

0.20%

8.44%

79.93%

1.708 "

11.54%

0.40%

0.13%

0.82%

8.44%

78.67%

1.651 "

10.51%

0.49%

0.28%

0.83%

7.56%

80.33%

1.498 "

11.14%

0.50%

0.20%

0.72%

7.81%

79.63%

1.244 "

11.31%

0.48%

0.15%

0.47%

7.75%

79.84%

1.481 "

11 . 10%

0.55%

0.68%

0.66%

8.31%

78.70%

1.409 "

11.30%

0.41%

0.63%

0.43%

7.81%

79.42%

1.329 "

10.60%

0.49%

0.20%

0.21%

7.63%

80.87%

1.725 "

11.28%

0.47%

0.31%

0.27%

7.56%

80.11%

1.723 "

10.45%

0.45%

0.17%

0.60%

8.06%

80.23%

1.541 "

10.94%

0.44%

0.53%

0.76%

7.56%

79.77%

1.141 "

10.44%

0.54%

0.10%

0.31%

7.81%

80.80%

11.08%

0.85%

0.28%

0.44%

8.25%

79.10%

0.958 "

10.51%

0.74%

0.12%

0.16%

7.81%

80.66%

0.892 "

10.49%

0.60%

0.30%

0.32%

8.00%

80.29%

0.788 "

9.99%

0.48%

0.94%

0.33%

8.06%

80.20%

10.06%

0.55%

0.71%

0.51%

8.13%

80.04%

1.238 "

9.21%

1.23%

0.80%

0.77%

8.44%

79.55%

1.175 "

9.19%

0.72%

0.87%

0.56%

8.94%

79.72%

1.533 "

9.32%

0.57%

0.52%

0.45%

8.75%

80.39%

1.179 "

9.55%

0.77%

0.91%

0.47%

8.38%

79.92%

1.429 "

10.37%

0.58%

0.16%

0.23%

8.38%

80.28%,

1.413 "

10.04%

0.72%

0.59%

0.45%

7.63%

80.57%

1.581 "

10.81%

0.51%

0.44%

0.31%

8.63%

79.30%

76 Original Communications: Eighth International [vol.

TABLE II
Restji-t or Analysis of Two Samples op South Carolina Rice

Sam-
ple
No.

Weight of
100 grains

Mois-
ture

Ash

Ether
Ex-
tract

Crude
Fibre

Pro-
teids

Starch
and Car-
bohy-
drates

29
30

2.241 grams
2.238 "

10.23%
9.01%

0.47%
0.37%

0.42%
0.21%

0.29%
0.36%

9.00%
8.13%

79.59%
81.92%

TABLE III

Results of Phosphate DBTBEMnsTATioNS on Twenty-Seven Samples of
Rice submitted to the Institute of Industrial Reseabch by the
Siamese Legation, Washington, D. C.

Sample
No.

%P20.

Sample
No.

%PaO.

Sample
No.

%P20.

0.22

0.31

0.39

0.32

0.30

0.23

0.41

0.20

0.21

0.39

0.28

0.21

0.42

0.26

0.30

0.58

0.31

0.49

0.24

0.26

0.35

0.22

0.30

0.35

0.34

South Carohna rice .

29
30

0.29
0.24

Interpretation of Results.

A careful inspection of the results shows, that all of the analy-
ses of the samples submitted compare favorably in respect to
nutrition value with the samples given under the World's Fair
report which includes typical rice analyses as quoted by various
authorities (see Appendix A). The results also for the most part
compare well with the analyses of the South Carolina rices
given in Table II. The phosphorous content of the imported

xvin] Congress of Applied Chemistry 77

samples (Table III) shows considerable variation ; in some cases
it corresponds to the average for milled white rice which is re-
ported to be about 0.25%; in other cases it is as high as is
usually shown in rices treated by the parboiling process. It
would appear that the white rices as represented in the twenty-
seven imported samples show on the average as high a nutrition
value as the white rices from other sources. The moisture con-
tent and weight per 100 grains is somewhat low in the imported
samples, for the reason stated above.

Interpretation of the Analytical Results in Relation to the Etiology
of Beri-Beri.

It has recently been claimed by Doctors Fraser and Stanton
of the Institute for Medical Research, Kuala Lumpor, that the
low phosphorous content of white milled rices is a predisposing
cause of beri-beri. (See "The Lancet" London) Vol. 176, p. 451
(1909). It is further stated by Doctors Fraser and Stanton that:
"From epidemilogical considerations and from experimental
evidence it appears that Siam rice is considerably more potent in
its beri-beri producing powers than Rangoon rice."

Opposed to the conclusions of Doctors Fraser and Stanton
stands the opinion of Dr. Hamilton Wright, former Director of
the Institute for Medical Research, Federated Malay States, an
eminent investigator of the Etiology and Pathology of Beri-beri.
Dr. Wright's published opinion, based on years of study and
clinical experimentation is quoted below :

(An inquiry into the Etiology and Pathology of Beri-beri.
Hamilton Wright, M. D., Studies from Institute for Medical
Research, Federated Malay States, Vol. 2, No. 1, p. 58 (363).
"The theory of the causation of beri-beri that fits the above
facts and all others observed in British Malaya is that beri-beri
is due to a specific organism which gains entraace to the body
via the mouth, that it develops and produces a toxin chiefly in
the pyloric end of the stomach and duodenum, aad that the
toxin, being absorbed, acts atrophically on the peripheral ter-
minations of the afferent and efferent neurones. Further, that
the specific organism escapes in the fseces and lodges in confined
places through accident or the careless personal habits of those

78 Original Communications: Eighth International [vol.

affected by the disorder, and that in the presence of congenial
meteorological, climatic and artificial conditions of close associa-
tion from overcrowding, the organism becomes virulent and,
gaining entrance to the healthy body in food, etc., contaminated
by it, gives rise to an attack of the disease. The fact that the
germ remains so closely focal can, I think, be explained by its
being at once destroyed by the action of direct simlight or that
the presence of CO2 or some other gas is necessary for its virile
development. It seems from my observations here that the
active stage of the organism in the body is between three and
four weeks. I base this estimation on the facts that the prelimi-
nary feeling of oppression in the epigastrium ceases at the end
of about three weeks, and that it is rare to find the lesion of the
gastric and intestinal mucose in cases of only six weeks' standing."

Conclusion.

As far as the results of analysis can be interpreted in the light
of the information at hand, there would appear to be no reason
why the white milled rices from one section of the world should
be held more responsible for mal-nutrition than similar rices
from other sections.

APPENDIX A.

EXTRACT FKOM BULLETIN NO. 13, TJ. S. DEPABTMENT OF AGRI-

cuLTtJBE. Division of Chemistet

Foods and Food Adulterants. Investigations made under
direction of H. W. Wiley, Chief Chemist. Part 9. Cereals and
Cereal Products, Washington, D. C, 1898.

Rice may reach the analyst in three different states, viz.:
unhuUed, hulled, and poUshed. He may also have occasion to
examine the broken fragments used in polishing and hulling, the
waste in manufacturing rice bran and other products. The
most important of these products in the present connection is the
polished rice as it is found in commerce, ready for preparation
as food. Rice is a cereal in which the starchy matters predomi-
nate, and in which there is a marked deficiency of proteids and

XVIIl]

Congress of Applied Chemistry

oils as compared with other standard cereals. The composition
of rice, as determined by the analysis of samples exhibited at
the World's Columbian Exposition, and by standard authori-
ties, is best shown in the table of maxima, minima, and means,
as in the case of the other cereals which have been mentioned.
In the following table the items marked I, II, and III, repre-
sent data obtained at the World's Columbian Exposition, while
the means of all the samples there analyzed are given in another
part of the table.

Table of Maxima, Minima, and Means of Constituents of Rice

Kinds and Nos. of samples

p
MS

1. Rice in the hull (for-

eign) :

Maxima

Minima

Means

2. Unpolished rice (for-

eign):

Maxima

Minima

Means

3. Polished rice (foreign) :

Maxima

Minima

Means

Mean composition of pol-
ished rice, etc., as
given by Jenkins and
Winton.
Polished rice (10 analy-

Grams

a3.250

b2.842

2.979

C2.826

C2.260

2.466

b2.633

al.560

2.132

Per
cent

bll.52

a9.03

9.88

C12.57

clO.92

11.88

bl3.15

ell. 82

12.34

Per

cent

b8.40

a8.23

8.32

cTO.60

o7.27

8.02

blO.33

c5.42

7.18

Per
cent

b2.04

al.44

1.71

Per
cent

bll.47
b9.45
10.62

C2.26

cl.62

1.96

cO.54

c0.04

0.26

cl.OO

cO.87

0.93

aO.56

aO.27

0.40

Per
cent

a4.66

b3.26

4.12

cl.22

cl.04

1.15

aO.65

aO.28

0.46

Per
cent

a65.70

a65.01

65.35

C77.34

c73.35

76.05

081.66

b75.62

79.36

Rice bran (5 analyses) . .
Rice hulls (3 analyses). .
Rice polished (4 analy

ses)

12.40
9.70
8.20

10.00

7.40

12.10

3.60

11.70

0.40

10.90

0.70

7.30

0.20

9.50

35.70

6.30

0.40
10.00
13.20

6.70

79.20
49.90
38.60

58.00

a Guatemala.

b Johore.

Japan.

80 Original Communications: Eighth International [vol.

Table of Maxima, Minima, and Means of Constituents of Rice. — Continued

Kinds and Nos. of samples

Per

cent

ecQt

cent

Mean composition of rice.

etc., as given by Ko-

nig.

Unhulled rice (3 anal-

yses)

11.99
12.58

6.48
6.73

1.65
1.88

6.48
1.53

3.33
0.82

70.07

Hulled rice (41 analyses)

76.46

Polished rice (9 analy-

ses)

12.52

7.52

0.84

0,48

0.64

78.00

Means of World Fair sam-

ples.

Unliulled rice (4 anal-

yses')

2.929

10.28

7.95

1.65

10.42

4.09

65.60

Unpolished rice (6 anal-

2.466

11.88

8.02

1.96

0.93

1.15

76.05

Polished rice (14 anal-

yses)

2.132

12.34

7.18

0.26

0.40

0.46

79.36

The mean composition of the different classes of rice as shown
by the analyses of the World's Fair samples is almost the same
as that shown by the work of other analysts collated as indicated
above. A tjrpical unhuUed rice has about the following composi-
tion:

Weight of 100 kernels, grams 3 . 00 Crude fiber, per cent 9 . 00

Moisture, per cent 10.50 Ash, per cent 4.00

Proteids, per cent 7 . 50 Carbohydrates, other than

Ether extract, per cent 1.60 crude fiber, per cent 67.40

A typical hulled rice, but unpolished, has about the following
compositions :

Weight of 100 kernels, grams 2.50 Crude fiber, per cent 1.00

Moisture, per cent 12.00 Ash, per cent 1.00

Proteids, per cent 8.00 Carbohydrates, other than

Ether extract, per cent 2.00 crude fiber, per cent 76.00

xviii] Congress of Applied Chemistry 81

A typical polished rice has a composition represented by the
following numbers :

Weight of 100 kernels, grams 2.20 Crude fiber, per cent 0.40

Moisture, per cent 12 . 40 Ash, per cent 0 . 50

Proteids, per cent 7.50 Carbohydrates, other than

Ether extract, per cent 0.40 crude fiber, per cent 78.80

SULLA MATURAZIONE DEL FORMAGGIO PECORINO

Prof. Dr. E. De' Conno
Delia R. Universitd, Napoli, Italy

II pecorino h un formaggio grasso costitiiito di sostanze azotate
e grasse; si ottiene dal latte di pecora, che ne pu6 dare fino al
22 % (fresco) = la cagliata si cuoce e poi si foggia in pani cilin-
drici che vengono salati e si fanno stagionare almeno per nove
mesi.

La sua pasta k bianco-gialliccia, omogenea o con piccoli e
scarsi occhi = ha sapore ed odore piccanti, come i latticini pecorini
in genere e tanto piil pronunziati quanto piil il formaggio 6
stagionato.

II costituente principale del formaggio h la caseina, la quale,
durante il periodo della maturazione, per eflfetto di special!
fermentazioni, origina materie albuminoid! solubili, ammidi
prodotti ammoniacali, altre sostanze di natura ancora non ben
definita e forse anche sostanze grasse.

II formaggio contiene, oltre la caseina, acqua, sostanze grasse,
lattosio, sali mineral! (NaCl) in proporzioni molto variabil! a
secondo della provenienza de! modi di fabbricazione e dell'etS,.

Lo studio delle trasformazioni che il formaggio subisce quando
h abbandonato all'azione delle diastasi e dei microrganismi, i
quali fanno ad esso subire un cambiamento completo, fornisce
uno de! piil important! capitoli della chimica del formaggio.

Questo prodotto durante la maturazione subisce a poco a poco
diverse modificazioni. La massa caseosa prende im aspetto
untuoso, nello stesso tempo che s! sviluppano I'odore e il sapore
che caratterizzano il formaggio mature, nel quale I'analisi indica
piccole quantity di ammoniaca, di acid! grassi e di leucina.

In principio, nel formaggio fresco v! sono necessariamente
gl! element! del latte = caseina, burro ed anche del lattosio che
la premitura non ha completamente eliminato.

Era interessante ricercare c!6 che divengono quest! princip!
durante la fermentazione casica. La perdita d! peso che la
materia subisce in seguito a questa fermentazione, perfettamente

84 Original Communications: Eighth International [vol.

constatata dall'esperienza 6 dovuta alia distruzione totale o
parziale dell'uno o dell'altro di quest! principi? Quali sono
quelli che resistono, quali quelli che spariscono? La materia
grassa h realmente aumentata nell'invecchiamento del formaggio?

In conseguenza di uno studio sul formaggio di Roquefort il
Blondeau (1) affermd che nella maturazione del formaggio si
formavano-dei principi grassi a spesa della caseina, la qual cosa
fu poi confermata da Keinmerick (2) e da Fleischer (3) nonchfe da
Musso e Menozzi (4), i quali credono appunto che la formazione del
grasso nelle stracchino abbia origine indirettamente dalla caseina.

II Brassier (5) intanto poco dopo del Blondeau, in uno studio
sulla trasformazione della caseina nella maturazione dello stesso
formaggio Roquefort, criticando fortemente le esperienze del
Blondeau, che egli dice "ben lontane d'essere al riparo di ogni
critica" afferma che non vi § formazione di grassi a spesS della
caseina; e gli studi di Manetti e Musso sul parmigiano confermano
questa opinione. Anche le ricerche del Kellner (6) sulla quantity
di burro in confronto di quella dell'acido fosforieo e della calce
confermano la stessa cosa, la quale provano pure le determina-
zioni di E. Schulze ed U. Weidmann (7).

Tali quistioni io ho voluto riesaminare analizzando il formaggio
in diverse epoche, cominciando dal momento in cui fu coagulate
e pressato.

Ho esaminato il pecorino da un punto di vista chimico, seguendo
le trasformazioni che la caseina subisce dal momento della coag-
ulazione fino a quello nel quale il formaggio maturo e messo
in commercio per la consumazione.

Si comprende facilmente che la maggiore difficolta consisteva
nel preparare un formaggio fresco di costituzione sufEcientemente
omogenea ed ecco come ho proceduto :

Del latte di pecora fu lasciato per 24 h. in cantina, indi, separa-
tane la crema, fu fatto agire il presame a 35°. Dopo la presa del

(1) Ann. de Chim. et de phys (4), I. 208.

(2) Pfluger's Aichiv. f. d. gesammte Physiologie (1869), 409.

(3) Virchow's Archiv. f. pathol. Anatomie u. phys. (1871), LI. 40.

(4) Le stazioni sperimentali agrarie italiane (1877), VI. 201.

(5) Ann. de Chim. et de phys (4), V, 270; C. (1865), 888.

(6) Landw. Versuchsst (1880), XXV. 39.

(7) Landw. Jahrbucher (1882), XI. 587.

xviii] Congress of Applied Chemistry 85

caglio fu favorito lo scolamento del siero mettendo a sgocciolare il
siero sopra una tela e sottomettendolo poi ad una forte pressione.

II formaggio aveva cosi una consistenza ferma e secca e fu
diviso in 15 parti che furono gettate sotto la pressa in apposite
forme.

II n. I esaminato immediatamente, ha date la composizione
del formaggio appena coagulate e pressato, e gli altri, che hanno
ricevuto la salatura con sale in polvere uniformemente ripartito
nella pasta prima di essere gittata in forma, furono esaminati
susseguentemente alia distanza di un mese I'uno dall'altro. Al
nono mese esaminando il formaggio n. 10 ho ottenuto risultati
uguali a quelli avuti per il n. 9 esaminato nell'ottavo mese, il
che mi indica che la maturazione si era completata all'ottavo
mese appunto e la prima parte del lavoro 6 cosi esaurita.

Resta ora a vedere se questa maturazione che all'ottavo mese
sembra completa, sia veramente tale o giunta a questo punto
continua molto piil lentamente, tanto che I'esame del formaggio
non d^ sensibili variazioni di composizione nel breve periodo
di un mese. Inoltre, dato che il processo continui, esso ha un
arresto definitivo o si collega con altro processo che trasforma
diversamente i prodotti formati nella maturazione? Mi riservo
di rispondere a cid con altra nota poich^ ho in corso esperienze
in proposito.

Ho creduto opportune esaminare, con le norme ordinarie,
il latte che 6 servito alia preparazione del formaggio nonch^
il prodotto secondario della preparazione stessa, il siero. Riporto
quindi qui appresso i risultati ottenuti rispettivamente dall'uno
e dall'altro esame.

COMPOSIZIONE CENTESIMALE DEL LATTE
Acqua gr: 81.2817

Grasso .
Caseina. . . .
Lattalbumina .
Lattosio ... .
Ceneri

6.8107
5.2716
1.0433
4.7106
0.8220

" 99.9399

86 Original Communications: Eighth International [vol.

COMPOSIZIONE CENTESIMALE DEL SIERRO

Acqua gr: 89.2449

Grasso " 2.9703

Sost. proteiche " 1 .7645

Lattosio " 4. 1619

Ceneri " 0.1670

Acidity (espressa in acido lattice). .. . " 0.0359

" 98.9945

EsAME Del Formaggio N. I.
(fresco = appena coagulato e pressato)

Caeatteki Fisici = Massa bianca, secca e fragile, senza odore
nd sapore sensibili. Messo su carta bibula non lascia traccia di
corpo grasso.

La carta di tornasole indica reazione leggermente acida.

Deteeminazione Di Acqua = La determinazione dell'acqua
fu fatta pesando esattamente una certa quantity, di formaggio e
seccandola prima nel vuoto su H2SO4 e poi in stufa a 110° fino
a peso costante. Poich^ il pecorino ha una pasta abbastanza
dura, ho creduto conveniente operare la determinazione diret-
tamente su di esso, senza aggiunta di sabbia, come generalmente
si usa fare con altri formaggi.

Per quanto, sia nel vuoto su H2SO4, che in stufa a 110°, vadano
via altre sostanze volatili, come NHs;. ed altri prodotti di decom-
posizione eventualmente presenti, pure la determinazione si
pu6 ritenere esatta, perche questi sono in tale piccola quantity
che si possono trascurare.

I risultati finali delle determinazioni sono liuniti nel seguente
quadro :

Prove H2O %

I gr: 40.7518

11 " 41.4391

HI " 41.0574

Media " 41.0828

XVIII

Congress of Applied Chemistry

Determinazione Delle Sostanze Grasse e Degli Acidi
Grassi = Questa determinazione fu fatta estraendo con etere
pure ed anidro il formaggio ridotto in piccoli pezzi e mantenuto
in sacchetto di carta in apparecchio Soxlet.

Poichfe I'etere estrae dal formaggio, insieme al grasso, Tacido
lattico e gli acidi grassi eventualmente presenti, cosi Testratto
etereo, prima seccato e pesato, fu poi ripreso con etere, neutraliz-
zando I'acidit^ con soluzione di carbonato sodico. — In tal modo
i gliceridi restano nella soluzione eterea, mentre passano in quella
acquosa, i saponi alcalini degli acidi grassi e I'acido lattico alio
stato di lattato.

Separata la soluzione eterea, e ripetutamente lavata con acqua,
fu determinato esattamente il contenuto in grasso neutro, dis-
tillando I'etere dalla soluzione eterea e pesando il residuo dopo
averlo seccato a 110°.

Per differenza dall'estratto etereo si ha la somma degli acidi
grassi liberi e dell'acido lattico (formati nella maturazione),
dalla quale somma togliendo la quantity di acido lattico, che si
determina a parte, si ha anche il contenuto in acidi grassi.

RISULTADO DELLE DETERMINAZIONI

Nel prodotto naturaJe

Grasso neutro
per 100 nel
prodotto sec-
cato a 110°

Prove

Estratto
etereo

Grasso

neutro

Ac. latt.

acidi grassi

III

2.7391
2.7902
2.7454

1.8583
1.8824
1.8750

0.8808
0.9078
0.8704

3.1540
3.1949
3.1824

Media

2.7582

1.8719

0.8863

3.1771

La materia grassa, ottenuta per evaporazione dell' etere, aveva
la pill grande analogia col burro per il suo sapore e per la temper-
atura di fusione che era 28°— Credetti opportuno trattarla con
soluzione alcoolica di KOH bollente per esaminare i prodotti

88 Original Communications: Eighth International [vol.

della sua saponificazione: ottenni, dopo saturazione dell'alcale
con HCl diluito, delle laminette cristalline che, disseccate in
carta bibula, abbandonarono a questa una certa quantity, di
sostanza oleosa, restando una materia che io potetti far cris-
tallizzare sciogliendola in alcool. Le laminstte brillanti, micacee,
che cosi ottenni, ricordavano le lamine dell'acido margarico.
La sostanza oleosa, della quale s'era impregnata la carta non
poteva essere altro che acido oleico; ma la quantity, di materia
sulla quale operavo, era troppo piccola per permettermi d'ac-
quistare una nozione dompleta sulla natura della materia che
presenta tutti i caratteri del burro. Era importante consta-
tare che la quantity di grasso che si trova nel formaggio appena
preparato non oltrepassa il 2 %, e che esso non pu6 essere che
del biurro meccanicamente, trasportato mella preparazione.

Deteeminazione Dell'Acidita Rifebita in Acido Lat-
Tico = La determinazione fu f atta nel prodotto naturale adoper-
ando circa gr : 10 di campione per ogni prova. 1 gr : 10 di sostanza
venivano scaldati con acqua a pill riprese, decantando ogni
volta il hquido: i liquidi riuniti e filtrati furon portati a 200 cc,
e sopra 100 cc. (circa gr: 5 di sostanza) fu titolata I'acidita con
soluzione N — 10 di KOH, indicatore il tornasole

I risultati sono rif eriti neH'unito quadro :

Nel prodotto naturale

Acidita rif erita
ad acido lattice,
% nel prodotto
seccato a 110°

Prove

Estratto
etereo

Grasso

neutro

Ac. latt.

ac. gras

I
II

0.7930
0.9106

1.3459
1.5455

Media

0.8518

0.8863

0.0345

1.4457

Deteeminazione del Lattosio = Questa determinazione non
ha grande importanza e non si esegue che nel formaggio freschis-
simo. Poich^ ^ difficile estrarre completamente il lattosio con
acqua leggermente scaldata, a circa gr: 50 di formaggio furono

XVIIl]

Congress of Applied Chemistry

aggiunti cc. 60 di una soluzione di NaOH diluitissima, legger-
mente scaldata, e in adatto recipiente fu continuato il mite
riscaldamento per qualche ora.

Freddata la massa grassa fu forata e se ne separd la soluzione
acquosa, ripetendo a piil riprese il trattamento.

I liquidi acquosi separati si acidificano con acido citrico per
precipitare la caseina e si filtrano. II filtrato si porta a volume
noto e su una parte aliquota di esso si precede direttamente al
dosamento del lattosio col liquido di Fehling.

Riporto nel seguente quadro i risultati ottenuti:

Prove

Lattosio %

Nel prodotto naturale

Nel prodotto seccato a
110

2.1913
2.2100
2.1752

3.7192

3.7510

Ill

3.6919

Media

2.1921

3.7207

Determinazione Dell'Azoto Totale = L'azoto totale fu
determinato col metodo Kyeldahl adoperando come ossidante,
insieme airH2S04 cone, un miscuglio di p.l di ossido di mercurio
giallo, p.l di solfato di rame e p.8 di solfato potassico. Distrutta
la materia organica, furon precipitati alio stato di solfuro il
mercurio ed il rame, prima della distillazione con ossido di mag-
nesio, mediante apposita soluzione di solfuro sodico. Anche questa
determinazione f u f atta sul prodotto naturale con i seguenti risultati :

Nel prodotto naturale

Nel prodotto seccato a 110°

Prove

N totale
%

Sostanze
azotate %

N totale

Sostanze
azotate %

II
III

8.5198
8.4913
8.5027

53.2487
53.0706
53.1418

14.2909
14.4122
14.4316

89.3181
90.0762
90.1975

Media

8.5046

53.1537

14.3782

89.8639

90 Original Communications: Eighth International [vol.

Detehminazione Delle CENEEi=La determinazione fu fatta
sul prodotto naturale. Riporto i risultati nel seguente quadro :

Ceneri %

Prove

Nel prodotto natur.

Nel prodotto seccato a
110°

0.7618
0.7392

1.2930

1.2546

Media

0.7505

1.2738

II pecorino dunque, appena coagulato e pressato ha la seguente
composizione centesimale media:

Nel prodotto
naturale

Nel prodotto
seccato a 110°

Acqua

41.0828
1.8719
0.8518
0.0345
2.1921

53.1537
0.7505

Sostanze grasse

3.1771

Acido lattico

1 4457

Altri acidi grassi

0.0585

Lattosio

3 7207

Sostanze azotate

89.8639

Ceneri

1 2738

99.9372

99.5397

Le varie determinazioni sui diversi formaggi, corrispondenti
ai n. 2, 3, 4, 5, 6, 7, 8, 9 e 10 furon fatte (due o tre prove per ogni
determinazione) seguendo i metodi precedentemente descritti
per il n. 1, aggiungendo I'esame delle sostanze azotate, nonch6
quelle suUa natura del grasso formato dopo che il formaggio
(n. 6) aveva acquistato tutti i caratteri esteriori che doveva
comunicare ad esso la stagionatura —

Come si sa, le sostanze azotate del formaggio sono costituite
da sostanze proteiche solubili ed insolubili, dai loro prodotti di
decomposizione, da saponi e sali ammoniacali.

xviii] Congress of Applied Chemistry 91

Questi diversi prodotti sono stati valutati nel modo seguente:

Ho pesato esattamente circa gr : 20 di formaggio, e, dopo averli
pestati in un mortaio, vi ho aggiunto acqua a poco a poco, fino
ad avere un volume uguale a circa il doppio di quello del campio-
ne — Ottenuta cosi una pasta omogenea, ho lasciato riposare per
le ^ h. perch6 si fossero imbevute bene di acqua tutte le particelle
solide, indi ho aggiunto ancora dell'acqua agitando — Separata
cosi la materia grassa, seguitai ad aggiungere acqua e versai il
tutto in palloncino tarato da 250 cc. fino a raggiungere il volume =
Agitai e lasciai riposare per 15 h. Chiarificatosi il liquido fu
filtrate, raccogliendone cc. 200.

Su cc. 25 di questo liquido fu fatta la determinazione di azoto
col solito metodo di Kyeldahl = Quest'azoto costituisce quello
dell'estratto acquoso, cio6 quello delle sostanze proteiche solubili
e dei prodotti di decomposizione (basi amidiche e ammoniaca),
che son pure solubili. Per differenza dall'azoto totale si ha
quello delle sostanze proteiche insolubili.

Questi due dati hanno una grande importanza in quanto danno
il peso ed il corrispondente valore in azoto della sostanza organica
resa solubile dai processi fermentativi, ed il loro rapporto dk
il coefficiente di maturazione —

Le sostanze proteiche sono state separate dai prodotti di
decomposizione (basi ammidiche) impiegando I'acido fosfo-
wolframico. La presenza di quest'acido non impedisce la determ-
inazione di azoto col metodo Kyeldahl nel precipitato e nel
liquido filtrate.

Cc. 50 del liquido precedentemente preparato furono portati a
cc. 70-80 aggiungendo cc. 15 di HCl al 20 % e precipitando con
fosfo-wolframato sodico. Raccolto sul filtro e lavato il precipi-
tato con soluzione d'H2S04 (5 %), poi con alcool assoluto, fu
seccato, determinando I'azoto sul prodotto secco. Dalla quan-
tity, d'azoto trovata fu tolta quella corrispondente airammoniaca
contenuta nella soluzione acquosa del formaggio, la quale h
pure precipitata dall'acido fosfo-wolframico, e la differenza
rappresentava quindi I'azoto delle sostanze proteiche solubili nelV-
acqua —

L'ammoniaca si determina a parte, su altri cc. 50 della soluzione
primitiva, spostandola, all 'eboUizione, con ossido di magnesio.

92 Original Communications: Eighth International [vol.

II rapporto fra I'azoto delle sostanze proteiche dell'estratto
acquoso a freddo e quelle totale del formaggio rappresenta 11
coefficiente di solubilizzazione, cioS I'azionedeifermenti diastasici. —

La differenza fra I'azoto totale dell'estratto acquoso e quelle
delle sostanze proteiche precipitate con I'acido fosfo-wolframico
costituisce I'azoto dei prodotti di decomposizione.

II rapporto fra I'azoto non proteico dell'estratto acquoso e
quelle totale del formaggio rappresenta il coefficiente di decom-
posizione, cic6 I'azione diretta dei microrganismi.

L'azoto ammidico 6 quelle che risulta dall'azoto totale dell es-
tratto acquoso detratte della somma dell'azote delle sostanze
proteiche selubili e dell'azoto ammoniacale.

Le basi ammidiche, provenienti dalla decomposizione della
caseina, sono essenzialmente costituite di leucina e contengono
solo in piccolissima quantitsl tirosina, butilammina, amilammina,
ecc. fine ad arrivare all'ammeniaca. Per questo nel fare il
calcole delle basi ammidiche daU'azoto trovato, ie he petuto
considerar questo come proveniente integralmente dalla leucina,
senza tema di grave errore.

Sperimentalmente infatti ho potuto esservare il seguente fatto:

II residue del trattamento con etere per I'estrazione delle
sostanze grasse e degh acidi grassi, ripreso con alcel a 36°, mi
forni, dope evaporazione del solvente, dei cristalli madraperlacei
di leucina, misti a piccelissime quantity di altri cerpi colorati
in gialle, che nen he esaminate sia per la scarsezza del materiale,
sia perche, alio scope del lavoro, la lore conoscenza non importava
gran che, potendosi dedurre appressimativamente la lore natura
da quanto finera si sa suUa maturaziene.

Cio che resta del formaggio, dope I'estrazione con etere e
relatives eccamente e cestituito da tutte le sostanze azotate
organiche; ammoniaca, lattosio e sali minerali, naturali ed ag-
giimti con la salatura. II residue cosi costituite cede all'alcool a
36°, quando venga trattato con questo solvente, tutti i suoi cesti-
tuenti ad eccezione della caseina: conoscende quindi il % di azoto
totale, nonche quelle ammoniacale, ed i % di lattosio e di saU
mineraU naturali ed aggixmti nel formaggio, ho determinate
I'azote totale dell'estratto alcoolco (36°), e sottraendo da esse
I'azoto ammoniacale, determinate a parte, he potuto conoscere

XVIIl]

Congress of Applied Chemistry

I'azoto ammidico, il quale corrisponde quasi esattamente a quelle
contenuto nella quantity, trovata di leucina.

ESAME DEL FORMAGGIO N. 2

^ (fresco di un mese che ha subito la salatura)

Caratteri Fi8ici = I1 formaggio che ha subito la salatura,
coagulate e pressato da un mese, ha completamente cambiato
d'aspetto. Esso ha gi^ preso I'aspetto d'un corpo grasso e mac-
chia la carta bibula sulla quale 6 deposto. II suo sapore comincia
ad esser dolce e piacevole, il suo odore appena sensibile.

COMPOSIZIONE CbNTESIMALE =

Nel prodotto
naturale

Nel prodotto
seccato a 110°

Acqua

Sostanze grasse

Acido lattice

Altri acidi grassi

Lattosio

SostaBze azotate

Ceneri e cloruro sodico

37.5080
7.9349
0.8649
0.7899
2.1949

44.5937
5.8099

12.7398
1.3887
1.2683
3.5177

71.5966
9.3280

99.6962

99.8391

EsME Sostanze Azotate =

Nel prodotto

naturale

Nel prodotto
seccato a 110°

Azoto

Sostanze
azotate

Azoto

Sostanze
azotate

Proteiche (ca^eina) |?°^"I''|^.,.

Insolubih

Non proteiche sol ( Ammoniacali .....

Amrmdiclie Leucrna

Totali

0.2529
6.5037
0.1436
0.2348
7.1350

1.5806

40.6481

0.1743

2.970

44.6000

0.4060

10.4418

0.2305

0.3769

11.4552

2.5375

65.2612

0.3198

3.5266

71.6451

94 Original Communications: Eighth International [vol.

I diversi coefficienti di solubilizzazione, decomposizione e
maturazione, calcolati da questi dati, come 6 precedentemente
detto, sono i seguenti:

CoEFFiciENTE DI SoLUBiLizzAziONE (dovuta all'azione

delle disastasi) 0 . 0354

CoEFFiciENTE DI Decomposizione (dovuta alia azione

dei microrganismi) 0 . 0530

COEFFICIENTE DI MaTURAZIONE 0 . 0970

Le sostanze azotate di questo formaggio hanno, secondo i
dati analitici, la seguente composizione centesimale:

Caseina solubile gr 3 . 5439

" insolubile " 91 .2289

Ammomaca " 0.3908

Basi ammidiche (leucina) " 4.9260

" 100.0896

Da questi risultati pare dimostrato che il soggiorno all' aria
(per la maturazione) del pecorino ha per effetto di aumentare
la quantity di materia grassa, che esso contiene in proporzione
abbastanza considerevole; ma prima di esaminare la natura del
grasso formato in queste condizioni, trovo conveniente attendere
che il formaggio abbia acquistato tutte le quality che un soggiorno
pill prolungato all'aria deve comunicargh.

La quantity di caseina invece, che prima era tutta insolubile,
6 considerevolmente diminuita, e di essa una parte, per azione
delle diastasi, si e resa solubile, porzione della quale, per azione
dei microrganismi, si 6 decomposta in basi ammidiche fino ad
arrivare aH'ammoniaca.

ESAME DEL FORMAGGIO N. 3

(fresco di due mesi che ha subito la salatura)

Caeatteri Fisici = Questo formaggio ha I'aspetto di un corpo
grasso come il precedents : ma il suo sapore 6 piil dolce e piacevole.

XVlIl]

Congress of Applied Chemistry

il suo odore piil sensible, propriety che vanno sempre piCl accen-
tuandosi nei formaggi corrispondenti ai successivi numeri 4, 5,
6, 7, 8, 9 e 10.

COMPOSIZIONE CeNTBSIMALE =

Nel prodotto
naturale

Nel prodotto
seccato a 110°

Acqua

Sostanze grasse

Acido lattico

Altri acidi grass!

Lattosio

Sostanze azotate

Ceneri e cloruro sodico

34.1019
13.9999
0.8799
1.5399
2.1949
40.3287
6.2819

99.3271

21.4644
1 .3491
2.3610
3.3653

61.8284
9.6314

99.9996

EsAMB Sostanze Azotate =

Nel prodotto
naturale

Nel prodotto

seccato a 110°

Azoto

Sostanze
azotate

Azoto

Sostanze
azotate

Proteiche (caaeina) 1?°^''!'^

[ Insolub

\T„ ™„* „„i J Ammoniacali

Non prot. sol. . -j- ,. /t • \
lAmmidiche (Leucina)

Total!

0.4888
5.2244
0.2799
0.4595
6.4526

3.0550

32.6525

0.3399

4.2340

40.2814

0.7494
8.0099
0.4291
0.7044
9.8928

4.6837

50.0618

0.5210

6.5910

61.8575

COEPFICIENTI =

di solubilizzazione 0 . 0757

di decomposizione 0 . 1145

di maturazione 0 • 2351

Original Communications: Eighth International [vol.

CoMPOSiziONE Centesimale Delle Sostanze Azotate =

Caseina solubile gr. 7 . 5841

" insolubile " 81.0609

Ammoniaca " 0 . 8438

Basi ammidiche (leucina) " 10.5110

99.9998

ESAME DEL FORMAGGIO N. 4
(salato di tre mesi)
CoMPOsizioNE Centesimale =

Nel prodotto
naturale

Nel prodotto
seccato a 110°

34.0178
21.4021
1.9968
1.9331
1.1198
33.0625
6.2115

Sostanze grasse

32.6375

3.0451

Altri acidi grassi

2.9480

Lattosio

1.7077

Sostanze azotate

50.1889

9.4724

99.7136

99.9996

EsAME Sostanze Azotate

Nel prodotto
naturale

Azoto

Sostazne
azotate

Nel prodotto

seecato a 110°

Azoto

Sostanze
azotate

Proteiohe (caseina) I ?°'^!''J^;,. •

i Insolubui

Nonprot.sol. 1 Ammoniacali

I Ammidiche (leucma)
TotaU

0.8823
3.4827
0.3530
0.5720
5.2900

5.5143

21.7668

0.4286

5.3522

33.0619

1.3393
5.2868
0.5358
0.8683
8.0902

8.3706

33.0425

0.6506

8.1246

50.1883

XVIIl]

Congress of Applied Chemistry

COEFFICIENTI

di solubilizzazione 0 . 1667

di decomposizione . . 0 . 1748

di maturazione 0 . 3415

CoMPOsizioNE Centesimale delle Sostanze Azotate:

Caseina solubile gr. 16.7089

" insolubile " 65.8365

Ammoniaca " 1 . 2963

Basi ammidiche (leucina) " 16. 1884

" 100.0301

ESAME DEL FORMAGGIO N. 5

(salato di quattro mesi)

CoMPOsizioNB Centesimale =

Nel prodotto
naturale

Nel prodotto
seccato a 110°

Acqua

Sostanze grasse

Acido lattioo

Altri acidi grassi

Lattosio

Sostanze azotate

Ceneri e cloruro sodico

33.9765
25.1742
2.5612
2.1375
0.5882
29.3750
6.2750

38.1293
3.8793
3.2375
0.8909

44.3583
9.5043

100.1276

99.9996

EsAMB Sostanze Azotate =

Nel prodotto
naturale

Nel prodotto

seccato a 110°

Azoto

Sostanze
azotate

Azoto

Sostanze
azotate

Proteiche (caseina) 1?°^"™

Insolub

v„„ « * if Ammoniacali

Non prot. sol. . •.■ . /> • \
[Ammidiche (leucma)

Totali

1.0180
2.6925
0.3897
0.5998
4.7000

6.3625

16.8281

0.4732

5.6187

29.2825

1.5372
4.0658
0.5884
0.9057
7.0971

9.6075

25.4112

0.7144

8.4746

44.2077

98 Original Communications: Eighth International [vol.

COEFFICIBNTI =

di solubilizzazione 0.2166

di decomposizione , 0 . 2105

di maturazione 0 . 7455

CoMPOsiziONE Centesimale delle Sostanze Azotate =

Caseina solubile gr. 21.7279

" insolubile " 57.4681

Ammoniaca " 1 . 6159

Basi ammidiche (leucina) " 19. 1879

99.9998

ESAME DEL FORMAGGIO N. 6
(salato di cinque mesi)
CoMPOsizioNE Centesimale =

Nel prodotto
naturale

Nel prodotto
seccato a 110°

Acqua

Sostanze grasse

Acido lattico

Altri acidi grassi

Lattosio

Sostanze azotate

Ceneri e cloruro sodico

33.9551
27.2007
2.8578
2.2502
0.3238
27.0625
6.2744

99.9245

41.1852
4.3272
3.4071
0.4904

41.0895
9.5003

99.9997

EsAME Sostanze Azotate

Nel prodotto
naturale %

Nel prodotto
seccato a 110° %

Azoto

Sostanze
azotate

Azoto

Sostanze
azotate

Proteiehe (caseina) | ^^""^'^

I Insolub

Non prot. sol. 1 AmmoniacaU

Ammidiche (leucina) .
TotaU

1.0165
2.2456
0.4079
0.6600
4.3300

6.3531

14.0350

0.4953

6.1756

27.0590

1.5434
3.4096
0.6193
1.0021
6.5744

9.6462

21.3100

0.7520

9.3766

41.0848

XVIIl]

Congress of Applied Chemistry

COEFFICIENTI =

di solubilizzazione 0 . 2347

di decomposizione 0 . 2466

di maturazione 0 . 9282

CoMPOSizioNE Centesimale delle Sostanze Azotate =

Caseina solubile gr. 23 . 4786

" insolubile " 51 . 8681

Ammoniaca " 1 . 8304

Basi ammidiche (leucina) " 22.8227

99.9998

ESAME DEL FORMAGGIO N. 7

(saJato di sei mesi)

CoMPOsizioNE Centesimale =

Nel prodotto
naturale

Nel prodotto
seccatoa 110°

Acqua

33.9884
28.1554
2.9998
2.3025
0.1906
25.9875
6.3289

Sostanze grasse

42.6265

Acido lattice

4.5417

Altri acidi grassi

3.4860

Lattosio

0.2886

Sostanze azotate

39.4741

Ceneri e cloruro sodico

9.5818

99.9131

99.9987

EsAMB Sostanze Azotate =

Nel prodotto
naturale %

Azoto

Sostanze
azotate

Nel prodotto
seccato a 100° %

Azoto

Sostanze
azotate

Proteiche (caseina) I .. 1 u-v

XT X 1 f Ammoniaca

Non prot. sol. i . -j- -l n ■ \
[ Ammidiche (leucma)

Totali

1.0234
2.0395
0.4172
0.6779
4.1580

6.3962

12.7468

0.5066

6.3431

25.9927

1.5545
3.0979
0.6337
1.0297
6.3158

9.7156

19.2223

0.7695

9.6349

39.3423

100 Original Communications: Eighth International [vol.

Coefficient! =

di solubilizzazione 0 . 2461

di decomposizione 0 . 2633

di maturazione 1 . 0386

CoMPOsiziONE Centesimale delle Sostanze Azotate =

Caseina solubile gr. 24 . 6076

" insolubile " 49.0339

Ammoniaca " 1 . 9490

Basi ammidiche (leucina) " 24.4033

ESAME DEL FORMAGGIO N. 8

(salato di sette mesi)

CoMPosizioNE Centesimale =

99.9938

Acqua

Sostanze grasse

Acido lattice

Altri acidi grassi

Lattosio

Sostanze azotate

Ceneri e cloruro sodico

Nel prodotto

naturale

seccato a 110°

33.9432

28.5344

43.1968

3.0729

4.6520

2.3287

3.5253

0.1245

0.1885

25.8062

39.2319

6.2814

9.5092

100.0913

100.3037

EsAME Sostanze Azotate =

Nel prodotto
naturale %

Nel prodotto
seccato a 110° %

AzOto

Sostanze
azotate

Azoto

Sostanze
azotate

Tj . ■ . / . SolubiU

Proteiche (casema ,. , ,

Insolub

Non Drot 1 I Ammoniacali

I Ammidiche (leucina) .
TotaH

1.0294
1.9948
0.4216
0.6832
4 1290

6.4337

12.4675

0.5119

6.3927

25 8058

1.5649
3.0376
0.6409
1.0386
6.2780

10.0797

18.9537

0.7782

9.7181

39.6297

XVIIl]

Congress of Applied Chemistry

101

COEFFICIENTI =

di solubilizzazione 0.2493

di decomposizione 0 . 2675

di maturazione 1 . 0698

CoMPOSiziONE Centesimale delle Sostanze Azotate =

Caseina solubile gr. 24 . 9312

" insolubile " 48.3127

Ammoniaca " 1 . 9836

Basi ammidiche (leucina) " 24.7723

ESAME DEL FORMAGGIO N. 9
(salato di otto mesi)
CoMPOsizioNE Centesimale =

99.9998

Nel prodotto
naturale

Nel prodotto
seccatoallO"

Acqua

SoBtanze grasse

Acido lattico

Altri acidi grassi

Lattosio

Sostanze azotate

Ceneri e cloniro sodico

33.9415
29.1974
3.1319
2.3399
0.0529
25.0000
6.2820

99.9456

44.0852
4.7579
3.5547
0.0805

37.9783
9.5433

99.9999

EsAME Sostanze Azotate =

Nel prodotto
naturale %

Azoto

Sostanze
azotate

Nel prodotto
seccatoallO" %

Azoto

Sostanze
azotate

Proteiche (caseina) j i^oi^b.;.'.'.'. '. ! ^

,, , , f Ammoniacali

Non prot. sol. j ^^jdiche (leucina).

Totali

1.0297
1.8586
0.4249
0.6868
4.0000

6.4356

11.6162

0.5159

6.4263

24.9940

1.5642
2.8234
0.6439
1.0433
6.0748

9.7762

17.6462

0.7818

9.7621

37.9663

102 Original Communications: Eighth International [vol.

COEFFICIENTI =

di solubilizzazione 0 . 2574

di decomposizione 0 . 2779

di maturazione 1 . 1521

CoMPOsizioNE Centesimale delle Sostanze Azotate =

Caseina solubile gr. 25 . 7485

" insolubile " 46.4759

Ammoniaca " 2.0640

Basi ammidiche (leucina) " 25.7113

ESAME DEL FORMAGGIO N. 10
(salato di nove mesi)
CoMPOSiziONE Centesimale =

99.9997

Nel prodotto
naturale

Nel prodotto
seccato a 110°

Acqua

Sostanze grasse

Acido lattico

Altri acidi grassi

Lattosio

Sostanze azotate

Ceneri e cloniro sodico

33.9421
29.1810
3.1504
2.3595
0.0573
25.0000
6.1531

44.1749
4.7693
3.5720
0.0868

38.0818
9.3148

99.8434

99.9996

EsAME Sostanze Azotate =

Nel prodotto
naturale %

Azoto

Sostanze
azotate

Nel prodotto
seccato a 110° %

Azoto

Sostanze
azotate

Tj , . , / . -, i Solubih

rroteiche (casema) i ^ , ,

\ Insolub

T.T J. 1 f AmmoniacaU

Nonprot. sol. | . . ,. , „ . ^
[ Amimdicne (leucma)

Total!

1.0346
1.8401
0.4264
0.6989
4.0000

6.4662

11.5006

0.5177

6.5396

25.0929

1.5759
2.8029
0.6495
1.0646
6.0929

9.8493

17.5181

0.7885

9.9614

38.1174

xviii] Congress of Applied Chemistry 103

COEFFICIENTI =

di solubilizzazione 0 . 2586

di decomposizione 0.2813

di maturazione 1 . 1737

CoMPOSiziONE Centesimale delle Sostanze Azotate =

Caseina solubile gr. 25 . 8398

" insolubile " 45.9580

Ammoniaca " 2 . 0688

Basi ammidiche (leucina) " 26. 1332

99.9998

Riassumo nei seguenti quadri i risultati analitici di tut.ti i
formaggi esaminati sostituendo alle sostanze azotate totali,
calcolate dall'azoto trovato, quelle risultanti dall'esame di esse.

104 Original Communications: Eighth International [vol.

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xvin]

Congress of Applied Chemistry

105

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106 Original Communications: Eighth International [vol.

Dal confronto delle precedent! analisi si vede chiaramente
che nella maturazione la sostanza grassa aumenta, rapidamente
nei primi mesi e molto piil lentamente in seguito con una per-
centuale che dal 2 % circa nel formaggio fresco si eleva fino ad
oltre il 29 % nel prodotto mature. In relazione a quest'aumento
h la diminuzione, per quanto non proporzionale, delle sostanze
azotate, le quali, diminuendo, mutano anche natura, poichS
si solubilizzano e si decompongono fino ad arrivare all'ammo-
niaca, prodotto che viene in parte fissato dagh acidi grassi pro-
venienti dall'ossidazione dell'oleina, costituente, come dimos-
trero in seguito, della sostanza grassa contenuta nel formaggio.

In diminuzione, per la stagionatura, 6 pure il lattosio; ma
proporzionalmente in aumento 6 I'acido lattice, il che mostra
la trasformazione del primo nel secondo prodotto.

Osservando poi la composizione centesimale delle sostanze
azotate aU'inizio ed a fine maturazione si vede come queste,
essenzialmente costituite in principio di caseina insolubile, vadano
gradualmente trasformandosi, tanto che a fine maturazione
la caseina insolubile non costituisce che il 46 % circa delle sostanze
azotate totali, mentre il resto risulta trasformato parte in caseina
solubile, parte in ammoniaca e basi ammidiche (leucina) con
preponderanza di queste ultime.

I coefficienti di solubilizzazione e di decomposizione, in relazione
con le trasformazione che man mano subiscono le sostanze azotate,
crescono entrambi, ma non proporzionatamente I'uno all'altro,
e mentre aU'inizio della maturazione il primo ha un valore numer-
ico di poco superiore alia met^ del secondo, alia fine diventano
quasi uguali mantenendosi sempre per6 una lievissima prevalenza
del secondo sul primo; La loro somma inoltre, che a principio
rappresenta quasi il coefficiente di maturazione, nel prodotto
stagionato 6 inferiore alia metk del valore numerico di quest'ul-
timo coefiiciente —

Da quest'osservazione si pu6 trarre un'utilissima conclusione
circa il giudizio sul grado di maturazione di un formaggio, e
cio6 UN Prodotto E'Tanto Piu'Matubo Quanto Piu'Vicini
Fka Loro Sono I Valori Dei Coefficienti di Solubilizza-
zione E Di Decomposizione (il primo deve sempre essere inferio-

xviii] Congress of Applied Chemistry 107

re al secondo) B Quanto Pitj'La Loro Somma E'Inferiore
Alla Meta'Del Coefficiente Di Mattjrazione.

Dietro i suddetti risultati dunque non si pu6 avere piil dubbio:
della materia grassa si h formata a spesa della caseina, durante
la maturazione.

Prima di discutere il modo come, io credo, questa materia
abbia potuto formarsi ho cercato di precisarne bene la natm-a.

II formaggio n. 6 (salato di cinque mesi) possedeva tutte le
qualitS, che esso era suscettibile di acquistare. Untuoso al gusto,
presentava, tagliato col coltello, un aspetto grasso. Esso non
si divideva piil in frammenti e macchiava la carta sulla quale
si deponeva. Possedeva inoltre un debole odore non notato nei
formaggi precedenti, e credetti quindi opportuno fare I'esame del
grasso in esso contenuto.

Per preparare la materia grassa da servire per I'esame chimico
ho adottato il metodo del Labor atorio Chimico di Washington,
che qui appresso descrivo:

Gr. 300 di formaggio, ridotti in frammenti della grandezza
di un pisello, si trattano con 700 cc. di potassa (50 %) a 20° in
una bottiglia a bocca larga, promuovendo la dissoluzione della
caseina con una forte agitazione.

In dieci minuti la caseina 6 sciolta, ed il grasso viene alia super-
ficie in piccole masse. Scuotendo il recipiente si fa in modo che
il grasso si radimi in una massa sola, e con I'aggiunta, nel recipient
di acqua fresca, il grasso raggiunge il collo del vaso, dal quale
si pu6 togliere con un cucchiaio. In tale operazione esso non
viene attaccato, e, lavato con molta acqua per asportare tutto
il residue non grasso che vi si pu6 trovare, in poco tempo 6 com-
pletamente separato, e si pu6 preparare, filtrandolo ed asciu-
gandolo, come 6 prescritto per I'esame delle materie grasse —

Questo grasso, che si trovava presente nel formaggio, nella
proporzione del 27, 2007 %, sciolto in alcool bollente, ha abban-
donato, per raffreddamento, dei piccoU cristalli di una sostanza
bianca, che, purificata per diversi trattamenti con alcool, ha
fornito dei cristalli di aspetto madraperlaceo, costituiti di mar-
garina pura: fondono infatti a 41°, e sottoposti all'azione di una
temperatura elevata si decompongono dando luogo ad acroleina.

108 Original Communications: Eighth International [vol.

Per la sua caratterizzazione ho eseguito sulla sostanza le
determinazioni seguenti coi risultatti che qui sotto trascrivo :

Indice di saponificazione 198 . 0

Indice di acidit£t 0.0

Numero degli eteri 198 . 0

" dijodio 0.0

" degli acidi volatili 0.0

" degli acidi fissi 95 . 51

Questi caratteri sono abbastanza precisi per provare che la
sostanza grassa, formata a spese della caseina, contiene della
margarina; ma per maggiore sicurezza, ed a conferma di cid,
ho sottoposto ad analisi la sostanza che ero riuscito ad isolare,
nonch6 I'acido grasso proveniente dalla sua saponificazione.

ANALISI DKLLA SOSTANZA GEASSA FUSIBILE A 41°

I. Gr. 0. 2962 di sostanza danno gr. O. 8284 di CO2 e gr. 0. 3269
diHaO.
II. Gr. 0. 3062 di sostanza danno gr. 0. 8562 di CO2 e gr. 0.
3387 di H2O.
E calcolando il % :

Trovato

Calcolato per
CitHioiOs

C=76.26
H = 12.26
0= ...

76.25
12.28

76.41
12.26
11.33

100.00

Per ottenere I'acido grasso corrispondente ho saponificato
con KOH ed ho poi decomposta I'emulsione saponosa con H2SO4
diluito. L'acido grasso, cosi messo in liberty,, si riunisce alia
superficie del liquido e comincia ad ammassarsi verso 40°. —
Questa sostanza, sciolta in alcool bollente, d^, per raffreddamento,
delle pagliette madraperlacee, che fondono a 58° — 60°.

xvin]

Congress of Applied Chemistry

109

ANAL18I DELL'aCIDO PUSIBILE A 59°

I. Gr. 0.3292 di sostanza danno gr. 0.9100 di CO2 e gr. 0.3736 di

HsO.
II. Gr. 0.1905 di sostanza danno gr. 0.5270 di COj e gr. 0.2159 di
H2O.
Calcolando il % :

Trovato

Calcolato per

CitHmOi

C = 75.38
H = 12.60
0= ...

75.43
12.58

75.55
12.59
11.86

100.00

La sostanza grassa contenuta nel pecorino, dietro questi
risultati, non 6 che margarina, accompagnata da un altro corpo
grasso, maggiormente solubile in alcool, che si ritrova nel solvente
dopo che si 6 deposta, per raffreddamento, la margarina.

L'alcool evaporato a h.m. ha lasciato, come residuo, un olio
leggermente giallastro, di sapore dolce e untuoso, liquido alia
temperatura ordinaria. Quest'olio, scaldato, si decompone a
260° dando luogo a vapori di acroleina.

Su di esso ho eseguito, come sul precedente grasso solido, le
seguenti determinazioni coi risultati che trascrivo :

Indice di saponificazione 190 . 0

" di acidity 00

Numero degli eteri 190 ■ 0

" di jodio 83 .93

" degli acidi volatili 169.68

" " " fissi 0.0

Sottoposto ad analisi tha dato i seguenti risultati:

I. Gr. 0.2341 di sostanza danno gr. 0.6621 di CO2 e gr. 0.2453

diHaO.
II. Gr. 0.2785 di sostanza danno gr. 0.7901 di CO2 e gr. 0.2914

di H2O.

110 Original Communications: Eighth International [vol.

III. Gr. 0.1975 di sostanza danno gr. 0.5602 di CO2 e gr. 0.2074
diHjO.
Calcolando il % :

Trovato

Caloolato per
C«H,„iO.

III

C = 77.12
H = 11.64
0= ...

77.36
11.62

77.35
11.66

77.37
11.76
10.87

100.00

Quantunque la propriety, e I'analisi dimostrino che la sostanza
liquida che accompagna la margarina non ^ che oleina, ho voluto
saponificarla, e fare I'anaUsi dell'acido grasso proveniente da
questa saponificazione.

L'olio ottenuto dall'evaporazione dell'alcool, dal quale si era
separata la margarina, 6 stato saponificato con soda e I'emulsione
decomposta con H2SO4 diluito — Si § cosi ottenuta una sostanza
bianca leggermente giallastra, di sapors dolce e untuoso, che
resta liquida fino a — 10°.

Sottoposta ad analisi ha dato i seguenti risultati :
I. Gr. 0.2202 di sostanza danno gr. 0.6170 di CO2 e gr. 0.2405

diHaO—
II. Gr. 0.2005 di sostanza danno gr. 0.5620 di CO2 e gr. 0.2182
di H2O.

Calcolando il % :

Trovato

Calcolato per

CisHitOa

C = 76.41
H = 12.11
0- ...

76.43
12.08

76.59 ,

12.05

11.36

100.00

xviii] Congress of Applied Chemistry 111

Questi risultati dimostrano che la sostanza grassa contenuta
nel pecorino fe un miscuglio di margarina ed oleina con prevalenza
della prima, giacchd dei gr. 27, 2007 di grasso contenuto in gr.
100 di formaggio, gr. 15, 4337 sono di margarina e gr — 11,7570
di oleina.

Dalle analisi precedenti si deduce che la caseina, nella matur-
azione, si h in parte trasformata in materia grassa, e cid che 6
evidente 6 la singolare coincidenza che la sostanza grassa del
burro 6 formata egualmente di margarina ed oleina presso a
poco nelle stesse proporzioni nelle quali queste due sostanze
costituiscono il grasso del formaggio, il che fa pensare che la
materia grassa del burro si sia formata, nell'economia, a spese
della caseina per reazioni analoghe a quelle che nel formaggio
han dato luogo alia sostanza grassa che ivi si trova.

I formaggi n. 7, 8, 9 e 10, specie questi ultimi — (9 e 10), con-
servati oltre il quinto mese, mostrano nei loro caratteri esteriori
delle profonde modificazioni. II colore non tarda molto ad
alterarsi, passa dal bianco al bruno, ed il formaggio prende un
odore sempre piil forte. Gambia anche il suo sapore e finisce
per acquistare un gusto forte e piccante.

Importava conoscere le cause di queste modificazioni, ed ho
voluto quindi esaminare anche il grasso del n.lO, come avevo
fatto per il n.6, poich^. avevo notato che dopo il quinto mese la
produzione di esso non aumentava piii sensibilmente, come nei
primi cinque mesi, pur rendendosi la percentuale delle sostanze
azotate sempre piil piccola.

La sostanza grassa nel formaggio n.lO e presente nella propor-
zione del 29 %. Di essa ho preparato un campione con lo stesso
metodo avanti descritto, e I'ho sottoposto alio stesso trattamento.
1 gr. 29 di sostanza grassa, di color giallo-carico, li ho trovati
costituiti di gr. 26.657 di margarina e di gr: 2,343 di oleina.

Questi risultati dimostrano che una parte del grasso che entra
nella costituzione del pecorino si e decomposta, mentre si^ e
formato del grasso nouvo, ed § sopratutto I'oleina che ha subito
una profonda modificazione a contatto dell'aria.

Bisognava quindi andare alia ricerca, separazione e dosamento

112 Original Communications: Eighth International [vol.

di questi prodotti di ossidazione dell'oleina e per far ci6 ho operato
nel modo seguente:

Ho scaldato gr. 500 di formaggio, ridotto a piccoli pezzi, con
circa 5 litri di acqua, a pill riprese, decantando ogni volta il
liquido=i liquidi riuniti e concentrati a un litro furon fatti
freddara e filtrati. II filtrato aveva preso una tinta giallastra e
possedeva inoltre un odore forte ed un sapore piccante che ricor-
dava quello del formaggio. Ho aggiunto al liquido acqua di
barite ed ho poi distillato : si 6 f ormato un abbondante precipitato
con sviluppo contemporaneo di odore ammoniacale. I vapori
sviluppati condensati erano alcalini e contenevano dell'ammon-
iaca, della quale ho constatato la reazione alcalina e I'effetto su
una bacchetta di vetro gabnata d'HCl.

L'ammoniaca era stata gi^ determinata nell'esame delle sos-
tanze azotate e non mi son quindi curato di determinarla ancora
qui: essa fu trovata in quailtit^, sufficiente per saturare tutti gli
acidi grassi presenti, pure a parte complessivamente determinati.

Ho evaporato dunque il liquido a piccolo volume, e durante
I'evaporazione si son depositati parte dei sali di bario che ho
cercato di separare gli unj dagli altri impiegando il metodo Lerch,
profittando cio6 della loro differente solubiUti in acqua calda —

Quando il liquido fu ridotto ad l/lO circa del volume primitivo
I'ho filtrato per separare i sali di bario depositati durante I'evapor-
azione, e nel filtrato si son prodotti, per il raffreddamento, degli
aghetti riuniti in fasci assai voluminosi del peso di gr: 5,9192;
i quali sottomessi ad analisi ban dato i seguenti risultati :

I. Grm: 0.4834 di sostanza danno gr. 0.6931 di CO2 e grm.

0.2663 di H2O
II. Grm: 0.4396 di sostanza danno gr. 0.6309 di CO2 e grm.
0.2430 di H2O

III. Grm: 0.9106 di sostanza fanno gr. 0.5773 di BaS04 corris-
pondente a gr. 0.33939 di Ba

IV. Gr: 0.8996 di sostanza danno gr: 0.5708 di BaS04 corrispon-
dente a gr. 0.33557 di Ba

Calcolando il % :

XVIUI

Congress of Applied Chemistry

113

Trovato

Calcolato per

Ba (PMn

0.).

III

C =39.09
H = 6.12

0 =

Ba=

39.13
6.14

37.27

37.30

39.23

5.99

17.46

37.32

100.00

Dai risultati di quest'analisi la sostanza solubile nell'acqua
boUente ridotta a piccolo volume era il sale di bario deU'acido
caproico, acido trovato nei prodotti d'ossidazione dell'acido
oleico.

Le acque madri non piil suscettibili di cristallizzazione le ho
trattate con H2SO4 ed ho ottenuto, per distillazione, un acido
incoloro che ricorda, per I'odore, il burro rancido, per cui ho pen-
sato che potesse essere dell'ossido butirrico, opinione che mi 6
stata confermata daH'analisi del suo sale di argento che ho
ottenuto precipitando una soluzione alcolica di AgNOs col liquido
acido raccolto nella distillazione.

II sale d'argento cosi ottenuto, lavato e seccato, pesava gr.

7.4432, il che corrisponde a gr. 3,3590 d'acido butirrico, acido

che entra nella composizione di questo sale cosi come lo dimostra

la seguente analisi.

I. Gr, 0.6292 di sostanza danno gr. 0.5665 di CO2, gr. 0.2056 di

HisO e gr. 0.3480 di Ag.
II. Gr. 0.4851 di sostanza danno gr. 0.4365 di CO2, gr. 0.1599 di
H2O e gr. 0.2684 di Ag.

Calcolando il % :

Trovato

Calcolato per

AgCHjO,

C =24.55
H = 3.63

0 =

Ag=55.30

24.53
3.66

55.32

24.61

3.58

16.43

55.38

100.00

114 Original Communications: Eighth International [vol.

Per questi dati sperimentali posso concludere che I'acido
butirrico e nel numero dei prodotti contenuti nel liquido filtrate.
I prodotti rimasti nel filtro I'ho addizionati con una certa
quantity di H2O e li ho messi a bollire, filtrando poi il^liquido
bollente. La mescolanza sulla quale operavo venne cosi divisa
in due parti = una solubile in H2O bollente, dalla quale la separai
per evaporazione a b.m., I'altra insolubile. Quest'ultima parte,
seccata e pesata, era gr. 3.2672 corrispondenti a gr. 2.3530 di
acido caprico, poiche il sale di bario sul quale operavo era del
caprato di bario come dimostra I'analisi:
I. Gr. 0.3564 di sostanza danno gr. 0.6527 di CO2 e gr. 0.2564

di H2O.
II. Gr. 0.3118 di sostanza danno gr. 0.5712 di CO2 e gr.
0.2246 di H2O.

III. Gr. 0.7115 di sostanza danno gr. 0.3456 di BaSOi corrispon-
denti a gr. 0.203178 di Ba.

IV. Gr. 0.7985 di sostanza danno gr. 0.3889 di BaSOi corri-
spondenti a gr. 0.2286 di Ba.

Calcolando il % :

Trovato

Calcolato per
Ba (CioHlOOj

III

C =49.94
H = 7.99

0 =

Ba=

49.95
8.00

28.55

28.63

50.10

7.93

13.37

28.60

100.00

Anche dell'acido caprico 6 stata notata da Redtenbacher la
presenza nei prodotti d'ossidazione, per NHO3, dell'acido oleico:

La parte solubile in H2O bollente fu ottenuta per evaporazione
del liquido: il suo peso era di gr: 3.2047.

Sottomessa ad analisi ha condotto ai seguenti risultati :

I. Gr. 0.3386 di sostanza danno gr. 0.5618 di CO2 e gr. 0.2186
di H2O.

xvni]

Congress of Applied Chemistry

115

II. Gr. 0.3058 di sostanza danno gr. 0.5079 di CO2 e gr. 0.1970
diHaO.

III. Gr. 0.7984 di sostanza danno gr. 0.4390 di BaS04 corris-
pondenti a gr. 0.25808 di Ba.

IV. Gr. 0.8125 di sostanza danno gr. 0.4472 di BaS04 corris-
pondenti a gr. 0.2629 di Ba.

Calcolando il % :

Trovato

Calcolato per
Ba (CB..O0.

III

C =45.24
H = 7.17

0 =

Ba=

45.29
7.15

32.32

32.35

45.39

7.09

15.14

32.38

100.00

I risultati di quest'analisi mi condussero ad ammettere che la
sostanza analizzata era il sale di bario dell'acido caprilico anch'esso
trovato nei prodotti d'ossidazione dell'acido oleico.

Riassumendo, i sali di bario che son riuscito a separare gli uni
dagli altri profittando della loro differente solubility in H2O
bollente, sono: Butirrato, Caproato, Caprato e Caprilato
DI Bario. Ho inoltre pesato questi differenti sali, i quali sono
presenti nelle seguenti proporzioni in gr. 500 di formaggio:

Butirrato d'argento gr. 7.4432 corrispondenti a gr. 3.3590 di ac.

butirrico
Caproato di bario gr. 5.9192 corrispondenti a gr. 3.7515 di ac.

caproico
Caprato di bario gr. 3.2672 corrispondenti a gr. 2.3530 di ac.

caproico
Caprilato di bario gr. 3.2047 corrispondenti a gr. 2.1820 di ac:

caprilico.

116 Original Communications: Eighth International [vol.

Calcolando il % in acidi liberi si ha:

Acido butirrico gr. 0 . 6718

" caproico " 0.7503

" caprico " 0.4706

" caprilico " 0.4364

" 2.3291

Dopo cio, poich^ nel formaggio (maturo) questi acidi sono
saturati dall'ammoniaca, si pu6 rappresentare la composizione
del formaggio n. 10 nella seguente maniera:

Acqua

Margarina

Oleina

Butirrato d'ammonio

Caproato "

Caprilato "

Caprato "

Acido lattico

Lattosio

Caseina solubile

" insolubile

Basi ammidiche (leucina) .......

Sostanze minerali (ceneri e Nacl)

• gr.

33.9421
26.6570
2.3430
0.8015
0.8602
0.4879
0.5752
3.1504
0.0573
6.4662
11.5006
6.5396
6.1581

99.5391

Sono dunque autorizzato a dire che nel pecorino conservato per
nove mesi a contatto dell' aria si trovano, indipendentemente dalla
margarina e dalla oleina, tutti i prodotti d'ossidazione di quest'ul-
tima sostanza, e poichS I'oleina che si trova nel formaggio di
cinque mesi 6 in buona parte sparita, bisogna concludere che gli
acidi butirrico, caprico, caprilico e caproico si originano dall'ossi-
dazione di questa sostanza.

Tutti questi acidi si trovano ugualmente nel burro invecchiato
con la differenza che nel formaggio essi sono saturati daU'ammoni-
aca, e sono appunto questi sali ammoniacali che danno al formag-
gio un sapore differente da quello del burro rancido, nel quale

xvin] Congress of Applied Chemistry 117

gli acidi sono gli stessi, ma alio stato libero, non satiirati da
alcuna base.

Nell'epoca in cui Chevreul intraprese il suo celebre lavoro sui
corpi grassi, egli si occupd dello studio del burro e trovd in questa
sostanza, divenuta rancida, gli acidi butirrico, caprico e caproico.
Malgrado lo stato imperfetto nel quale si trovava in quell'epoca
I'analisi organica egli seppe perfettamente distinguere questi
acidi e ne face anche uno studio abbastanza complete. Dopo
d'allora un gran numero di scienziati si 6 occupato dello stesso
argomento e ricordo specialmente Lerch, il quale dopo aver sap-
onificato il burro rancido lo distill6 con un eccesso d'HjSO*
diluito, guingendo cosi ad ottenere fino a cinque acidi volatili
(butirrico, caproico, caprico, caprilico e vaccinico).

M. Bromeis' ha studiato egualmente la costituzione del
burro e vi ha trovato, indipendentemente dalla margarina e
dall'oleina, dell'acido butirrico.

Dall'accordo di questi risultati con quelli che io stesso ho otte-
nuto si vede chiaramente che bisogna attribuire all'oleina I'irran-
cidimento del burro, come indubbiamente 6 all'ossidazione di
questa sostanza che bisogna attribuire la produzione dei differenti
acidi dei quali ho potuto constatare la presenza nel pecorino.
La sola differenza che sembra esistere h che nel burro questi acidi
sono alio stato libero, mentre nel formaggio essi sono combinati
all'ammoniaca.

Credo cosi d'aver dimostrato che la caseina si trasforma, nella
maturazione del formaggio, in una sostanza grassa avente la
piCl grande analogia col burro, poich6 si compone di margarina
ed oleina, e queste sostanze entrano nella sua costituzione presso
a poco nelle stesse proporzioni nelle quali esse si trovano nel

burro.

Resta ora a stabilire come questa trasformazione awiene,
ci6 che mi propongo di fare in una prossima nota.

Napoli, Istituto di Chimica Generale della R. University—
Maggio del 1912.

1 Ann. der Chem. und Phaxm. XLII, 46.

THE GRINDING OF CORN-MEAL FOR BREAD

By F. p. Dunnington
University of Virginia, Charlottesville, Va.

At the present time, when the high prices of food occasion so
much concern, and a conservation of all the resources of this
country awakens so much interest, it is somewhat amazing that
the United States produces such an enormous crop of Indian
Corn and yet in the larger portion of this Country it is consumed
in the form of bread to a very small extent.

Dr. Charles D. Woods, Director of the Maine Agricultural
Experiment Station, has compiled an excellent treatise on the
Food Value of Corn and Corn Products, published as Farmers'
Bulletin No. 298 by the U. S. Department of Agriculture 1907;
and in this he sets forth most plainly the advantages of compo-
sition, digestibility, wholesomeness, convenience and pecuniary
economy of corn as a food for man.

The Encyclopedia Britannica 11th Ed. p. 449 states: "As an
article of food maize is one of the most extensively used grains
of the world. Although rich in nitrogenous matter and fat it
does not make good bread."

It is generally understood, and so far as I have been able to
obtain reports, it appears that in the U. S. the considerable use
of corn as a bread is confined to the southern States and there
largely to the population of the coimtry, and smaller towns. In
many of these localities it is more largely used than is wheat.

The readiness with which it may be prepared and the rapid
and simple methods by which it may be cooked, as well as the
pleasant and satisfjdng character of the food, its composition
approaching to that of a complete ration, have much effect in
determining its uses as a staple food.

On the other hand, a considerable amount of corn meal is made
throughout the Central and Western States, especially at the

119

. 120 Original Communications: Eighth International [vol.

larger business centres — but it is used only sparingly; — often in
admixture with wheat flour and largely in the form of mush (or
hasty pudding). A few estimate its use as one fiftieth, and more
at one himdredth of that of wheat flour.

In 1890 the U. S. Secretary of Agriculture, Hon. J. M. Rusk,
endeavored to induce a larger use of corn a as bread stuff in
Europe and made some expenditures under the efficient man-
agement of Mr. Chas. J. Murphy as a Special Agent, but does
not seem to have succeeded, and there is probably even a smaller
proportional consumption now in the United States than there
was at that date in the form of bread, while the manufacture of
grits and other corn products has been largely increased.

It is my endeavor in this study to ascertain, why this apparent
inconsistency; that this cheaper, healthy food is so sparingly con-
sumed, where economy in living is of so great import.

The question seems narrowed down to the manner of making
the meal as being the factor which determines the extent of its
use; and hence this, the narrow range of the discussion in this
paper.

As to the grain itself, the comparatively high fat content, viz.,
5 per cent, constitutes a considerable portion of its food value,
while its presence adds to the care required in curing and keep-
ing the grain, and also to the difficulty of keeping the meal for
more than three or four weeks in most climates.

Hence it is that those supplying distant trade arid, most of the
larger mills, find it best to kiln-dry the grain, so destroying any
bacteria which may have infected it, and in grinding it, to re-
move the germ, so as to obtain a product freed in large measure
from these drawbacks, thus treating both yellow and white corn.

The grinding is conducted by water, steam, or electricity for
power, and we might judge their use as entirely a matter of indif-
ference, yet that is not wholly so, as I shall have occasion to men-
tion later on.

The mills employed formerly were only burr-stones (or occa-
sionally made of some local sandstone), but in recent years, most
of the larger mills use steel rollers, similar to those used for wheat
flour.

Samples of meal have been secured from most of the corn pro-

xvni] Congress of Applied Chemistry 121

ducing states, and to avoid advertising or embarassments, these
are designated by numbers, stating only the states from which
they come. In the endeavor to ascertain the different forms of
meal made, this collection of samples must not be considered as
representing the relative production, inasmuch as the very num-
erous small "custom mills" scattered throughout the rural dis-
tricts generally make corn meal in one very simple manner, with
a pair of burr-stones, driven slowly by water power.

The examination of these meals has been made as follows:
and since in bread making, any husk is always removed, all was
first passed through a sieve of sixteen meshes to the inch .

1st. Volume: 50 grms. of meal were jarred vertically, for 5
minutes, in a glass cylinder, 100 c.c. measuring 7 inches in height,
and from the volvune read off, the weight in pounds, of a bushel
of meal was obtained.

2nd. Site: 100 grms. of meal was shaken uniformly for 10
minutes in a nest of eight brass sieves, which time was longer
than necessary for most samples, but in a few instances, where
the sample was more oily, the smaller sizes were not sharply
separated. These sieves, to one inch, had meshes: 16, 20, 24,
30, 36, 40, 50. Any finer sizes would have been of no use. On
Table I, the per cent passing each number of mesh is indicated,
omitting fractions.

3rd. Fats: In some typical samples only, as indicated, a deter-
mination of the fat was made. Sudan III was used as a staining
in examining the meal under the microscope and proved very
satisfactory in bringing out the fat globules, but it afforded no
quantitative estimation. A comparative estimate of the amount
of freed fat was obtained by noting the length of time respec-
tively taken by each specimen to attain the same (dark brown)
color when soaking in a water solution of Osmic acid. About
.25 grms of meal in a watch glass was moistened by 3 c.c. of a
0.2 per cent solution of OsOs in from 10 to 30 minutes or more,
the uniform color was obtained. The comparison was made
with three pieces of cardboard giving near shades of a brownish
black color and the time was noted as the darkening sample
passed each of these shades of color; the average of these three
periods is the figure given on Table I.

122 Original Communications: Eighth International [vol.

And we may take the reciprocals of these figures as expressing
the amounts of free fat present.

4th. 'Cooking: While it may be that for each variety of meal
there is a mode of cooking to which it is specially adapted — in
order to obtain a comparison of the meals, the method of cook-
ing selected is of the simplest kind and one which conspicuously
brings out any imperfections or flavors.

Each sample was treated approximately as follows:

A. Meal, sifted 100 grms.

Salt 2 "

Water— about 90 "

Mixed quickly to a soft dough, formed to 3 or 4 small rolls,
placed on a pan and baked in a hot oven, at 440 Fahr. for 30
minutes.

B. Meal, sifted 100 grms.

Salt 2

Lard 10 "

Water — about 85 "

Mix with salt, "lightly" mix with lard— then with water and
bake as above.

One can expect httle satisfaction from single tests of this kind
not only because of the special difficulty of making ordinarily
good bread upon so small a scale, and of baking it uniformly
from day to day, but still more, in observing and expressing the
slight differences of taste that are presented.

In describing the character of the bread, the following abbre-
viations are employed in the Table:

g, good; p, poor; f, flavor; n, no flavor; s, sweet;
c, coars§; t, tough; d, dry; v, very;

w, white; W, exceptionally white; y, yellow; Y briUiant
yellow.

XVIIl]

Congress of Applied Chemistry

123

TABLE I.

Whole grain

■si

Sized by sieves mesh to inch

Kequired
color
min.

Cooked

burr ground

Color

Taste,
etc.

(1) Va.

(2) "

4.7

(3) "

7.4

(4) "

(5) "

vgfs

fat: 4.71

(6) Tenn.

1.6

4.4

(7) N. C.

3.3

vgf

(8) "

gfs

(9) Ala.

gst

fat: 4.39

(10) Del.

3.6

(11) Ark.

(12) Miss.

RoUer

ground

(13) Md.

(14) "

gft

(15) Ky.

gcs

(16) "

1.3

4.4

7.3

gcs

Fat: 2.20

(17) Mich.

gsf

(18) Iowa

7.4

24.

scf

(19) Kan.

3.7

vgf

Fat: 1.78

(20) Conn.

8.6

3.2

108

gcf

Fat: 1.75

Degermi-

nated

roller

ground

(21) Pa.

gcf

(22) Ky.

vgn

(23) Mo.

5.8

gen

(24) Mo.

Fat: 1.25

124 Original Communications: Eighth International [vol.

TABLE I

— Continued

Sized by aeves meah to inch

Hsquiied

Cooked

Whole grain

for
color

burr ground

min.

Color

etc.

(25) Iowa

(26) Neb.

gto

(27) lU.

(28) Mich.

4.7

g*f

Whole grain ground

at Univ. Va.

(29) lU. Agr. Dept.

fat: 5.66

(30) lU. Agr. Dept.

gnd

fat : low ground

(31) 111. Boone Co.

once:

(32) 111. Boone Co.

twice:

(33) lU. Boone Co.

thrice:

In order to compare the weights per bushel, it seemed well to
employ a uniform method of settling the meal, hence it was jarred
in the measure until it would settle no further. Specimens
containing much of fine powder settle very compact, therefore
in this respect these results cannot be compared with those usu-
ally observed.

It may be noted that some of the granular meals are compact
but others as (16) and (22) .which appear to be "cut meals" are
more bulky.

Some years ago, it was the custom of many steam driven mills
to grind corn by burr to a fine powder, in order to make a whiter
meal, resembling wheat flour; this made a very compact bread.
But no such meal is found among the samples examined, and
the making of such meal seems to be now generally discontinued
and replaced by the manufacture of a meal which is chiefly coarse
and contains but little of powder. The superior whiteness of

xvin] Congress of Applied Chemistry 125

some of these specimens is attained by selection of well-matured
white corn, "scouring" the grain when shelled, and removing
all husk and germ as soon as they are set free by the rollers em-
ployed— as seen in Nos. (19), (23), (25). Similarly from yellow
grain very clean specimens are Nos. (20) and (21).

It is to be noted that the nutty flavor of white corn (some-
what like that of a chestnut), as well as the peculiar flavor of
yellow corn are volatile bodies (i. e. odours) and are largely re-
tained by the fat. The flavor is therefore to a large ejttent re-
moved with the germ and is diminished by too much heating in
the grinding or by Kiln drying. It is also removed by too long
repeated grinding as is shown in No. 31 made by a single grind-
ing of good grain in a metal hand grist mill, while the same grain
was ground twice in making No. 32.

Nos. (29) and (30) were made of good corn and were run
through the metal grist mill three times, thereby losing all
"flavor." All the degerminated meals are found to be without
the nutty flavor although No. (22) is certainly excellent in all
other respects.

As to the kiln-dried meal of white corn, they too, generally
retain little flavor, but Nos. (19) and (22) make excellent bread
and have a pleasant sweet taste.

In examining this lot of selected samples, generally donated
by the makers, it is natural that in tasting the raw meal I found
but one that possessed any mustiness, and in this instance the
sample was obtained from a retail grocery store without any
inquiry as to its freshness.

The removal of the germ presents an important economical
feature. Taking the whole meal as containing 5 per cent, fat,
the degerminated meal will have about 1.5 per cent, fat, thus
100 lbs. of meal would so lose about 3.5 lbs. of edible digestible
fat, which will correspond, allowing for the increase in starch,
to a loss in food value of 30 cents or more. The reality of this
loss is brought out by the fact that when cooking the meal, or
other material to be eaten with it, one will ordinarily add this
much additional fat, at a cost of 10 cents or more per pound, to
replace that which has been removed.

As to the effect upon cooking.

126 Original Communications: Eighth International [vol.

The making of a proper dough largely depends upon the fine
particles retaining water sufficient to fully soften the enclosed
larger grains when heated in the oven, hot from the first. If
the meal is too largely coarse it can hardly be made to stick to-
gether, and the dough presents a rough surface, from which the
water may escape (dry out) before the starch and protein are
properly softened, so producing a very hard bread, insufficiently
cooked. An apphcation of this principle is presented in a favor-
ite method of cooking an "ash cake." The plain corn meal dough
is wrapped in a cabbage leaf and buried in hot ashes until done.
In this the flavor of the meal is peculiarly well preserved and
we may obtain a very appetising food. Hence it is that some
portion of the corn must be finely ground to obtain a meal for
general use.

If a meal is too fine, it may become too compact when made
into a dough, in such case the addition of a very little baking
powder, say 3 grms. to the foregoing receipt will sufficiently
open the dough to give good bread.

On the other hand very coarse and gritty meal is, in this re-
spect, better adapted to making mush or a batter bread in which,
while being cooked, it necessarily remains in contact with an
excess of water.

All of .the above samples of meal were also cooked according
to receipt B. and there was little variation in the good bread
obtained — tender, porous and appetising but varying with the
peculiarities of the grain as to texture, color, and flavor of the
meal.

The meal thus mixed with the fat should be but lightly pressed,
with no pressing or "working" (as must also be done in making
pastry with wheat flour), so subdividing the dough; and the re-
sulting bread will be excellent.

We have in corn a natural mixture of starch, protein, fibre
and fat, which, when simply ground and moistened by water,
gives a dough all ready for baking, the oil serving to separate
the mass and prevent it from becoming too compact or hard;
hence it is to be treated in a wholly different manner from wheat
and, to the writer, is not to be cooked with wheat flour with any
advantage.

xvin] Congress of Applied Chemistry 127

It seems therefore that the work done in refining corn meal
is, so far as its use for bread is concerned, not well directed, in
that the portion of the grain so removed is the very portion' in
which rests one of its chief advantages. It is true that meal
from whole corn will not keep, under most conditions, more than
3 or 4 weeks, but is it not profitable to supply this fresh meal to
obtain its several advantages?

In recent years there has been much and successful endeavor
to free wheat flour from all bran and husk, so securing its pure
whiteness while losing the advantages of the rougher fibre and
ash content. But I think I have shown that similar, processes
do not improve corn meal, and that there is by this process loss
of some of its most valuable constituents.

It appears from the favor with which oatmeal and many of
the numerous breakfast foods are received (so much so, that in
some cases these are sold at from 3 to 5 fold of the price of the
grain from which they are made), that there is a craving of many
stomachs for rougher food such as stimulates the processes of
digestion; and this is certainly in many instances the explana-
tion of the satisfaction with which corn bread is preferred by
many as a staple diet.

But when corn is well matiu'ed, kept to thoroughly dry on
the ear, and then, as it is needed, ground without heating, by
burr stones, slowly turned by a water wheel, it furnishes a sweet,
nutty flavored meal, which combines the most valuable of nutri-
ents, and when cooked in the simplest manner, furnishes a food
which is to many of mankind very acceptable, and to some, the
staple of Ufe.

(.Abstract:)

FOOD STANDARDS, THEIR NATURE, HISTORY, AND

FUNCTIONS

By William Fbear
State College, Pa.

Nearly all civilized lands have enacted general food laws cover-
ing all foods. They prohibit the sale, as normal, of products that
depart in certain, very generally defined ways from the normals
corresponding to the food names used; but do not define these
normals. In the absence of sufficiently complete, accurate, and
concise definitions of these normals, the executive officers of these
laws have been obUged to judge for themselves what the respec-
tive normals are, subject to the confirmation of the Courts.

A food standard is the expression of a food normal, and may
include chemical and physical limits indicative of kind and
quahty. The existing systems of standards, British, Bavarian,
German, Austrian, American, Swiss, Italian and Holland, are the
work of experts representing the executive branches of the
respective governments.

Nature: Food standards should correspond to the people's
concepts corresponding to the several food names, and, since the
laws with which they are to be used are quasi-criminal, should
correspond to the lowest quality, within the kind, acceptable
without notice of inferiority. They should represent present
usage, the concepts of the public, where they differ from those of
the trade, and those of home, instead of foreign coimtries. Since
foods are chiefly of domestic, rather than of factory production,
domestic usage should determine the normals, but due considera-
tion should be given to the requirements of commercial dis-
tribution, as contrasted with immediate, domestic consumption.

Matter: As their material, food standards should contain
what is necessary, (1) to distinguish the various normals from

129

130 Original Communications: Eighth International [vol.

each other; (2) to distinguish each normal from its substitutes,
imitations, and adulterated modifications. The data should be
both authentic and broadly representative. A standard may be
useful, even though incomplete; but incompleteness of chemical
and physical data should not lead to a definition of species broader
than the public concept therefor. Such data should represent
the products of the country in which the standard is to be used.

Consistency: Systems of food standards should be consistent;
but the criterion of consistency is external, not internal; con-
sistency with the people's concepts, not etymological consistency
is the aim. Even a given word varies in its meaning with the
context. Moreover, there is no simple mathematical formula
that can serve safely to fix the relation of the minimum to the
average of quality.

Form: A clear, concise definition, closely knit with the most
serviceable Umits, will better endure legal analysis than the
encyclopediac form of description with loosely appended chemi-
cal limits. The latter form is, however, superior educationally.

Function: Standards are practically essential as bases of
reference in the enforcement of general food laws, unify executive
and judicial decisions, reheve the trade and the public from
confusion and uncertainty. They do not interfere with variety
in production, nor should minimum standards tend to lower the
general average of excellence; since, however, such standards do
not in reality represent the absolute minima of quality, their
limits should be set with caution and common sense, and be
widely published before appUcation, lest honest, but vmskilled
or ignorant producers be injured.

International standards are practicable only for a limited num-
ber of products, but are desirable for these.

Provision should be made for the continuous addition of new
limits and standards as the need and data therefor appear; but
radical changes, made with frequency in existing standards, are
gravely unsettling to producing interests.

THE PACKING OF AMERICAN SARDINES
By H. H. Hanson

Scattered along the Maine coast from Portland to Eastport
there are about fifty-five different factories, whose combined
annual output is usually somewhere between 125 and 200 mil-
lions of cans of sardines, valued at from five to seven million
dollars according to the season. A large proportion of this
industry is located around Eastport and Lubec, in Passama-
quoddy Bay, where a majority of the inhabitants are de-
pendent upon it, either directly or indirectly, for their livelihood.

A study, not yet entirely completed, was undertaken for the
purpose of obtaining information on certain points of this
industry which were perplexing alike to the packers themselves
and to the food officials of the country. A comparison of the
American and Foreign packed sardines and a description of the
packing process is at once interesting and instructive as throwing
light on some of the points under consideration.

The name sardine, which comes from the island of Sardinia,
around which sardines abound, is not the name of a particular
species, but is applied to fish of the genus Clupea, various species
of which are canned in different parts of the world.

There are three important respects in which the Maine sar-
dines differ from the foreign sardines, of which the French pack
is generally recognized in this country as the most desirable.
First, the fish packed in France under the name sardine is the
Clupea ■pilchardus, while the fish packed in Maine under that
name is the Clupea harengus, two distinct species of the same
family which differ somewhat from each other both in appear-
ance and flavor. Second, French sardines are packed in olive
oil while the Maine sardines are put up in cottonseed oil. Third,
in handling the French pack the single fish is the unit and quahty
is at all times considered of paramount importance; while in
handling the Maine pack the hogshead is the unit and quantity
is always sought. In America the French sardine retails for from

131

132 Original Communications: Eighth International [vol.

thirty-five to sixty cents per can, while the Maine sardine retails
for the most part for five cents. The markets for these two gen-
eral grades seem to be well established and although some fancy
goods, which bring a high price, are put out it is quite certain
that the Maine packers cannot be brought to the point of adopt-
ing French methods of handling even though the product might
thus be improved in quality.

The fish are caught almost entirely in weirs, large circular or
oval traps having long wings extending out from the one opening
and arranged so as to guide the schools into it. These weirs are
large enclosures built mainly of brush topped with marlin and
are so located that there will be perhaps twenty feet of water in
them at low tide. Fish may be taken coming in on the flood
tide or going out on the ebb. It is not uncommon to take 100
hogsheads in a single weir, and even larger catches have at
times been reported.

When a catch has been made the weir-man closes the gate of
his trap and awaits the coming of the sardine boat. The com-
petition between the boats ruiming for the different factories is
often very keen. Not many years ago there was sharp bidding
between the skippers of these boats so that the prices paid for
the fish were often ridiculously out of proportion to their value.
As high as 130 per hogshead has been given. During the last
few years a more or less fixed price has been paid for the fish
and the boat which arrives first at the weir has first claim. At
the beginning of the season about $12 per hogshead may be
offered and later, when the fish become more plenty, the price
is dropped to about $6.

When the fish have been purchased a purse seine or net is
carried around the entire catch in the weir, drawn together at
the ends, and closed at the bottoiji, so that the sardines are
brougth into a compact mass, and may be scooped with hand
nets into dories and conveyed to the sardine boats lying just
outside. These boats range in size from remodelled fife boats
holding perhaps ten hogsheads to trim two-masters holding
eighty hogsheads. As the fish are scooped into the holds of
these boats salt is sprinkled over them, at the rate of about
100 pounds to the hogshead, and when the hold is full a race

xviii] Congress of Applied Chemistry 133

for the factory is begun, each skipper striving to land his catch
first. Arriving at the wharf of the factory the boat is drawn
under a windlass from which a tub is let down to the hold and
the fish are scooped into the tub, drawn up, and dumped into a
sluice as fast as it is possible to work. Along this sluice, flushed
with water, the fish run, sometimes through a contrivance
which separates the large from the small, into large wooden
pickUng tanks where they are kept in salt brine long enough to
give them a flavor and aid in preserving and hardening the
flesh. Time allowed in the pickle varies with the condition of
the fish and amount of salt they have already had in the boat.

From the tank most of the fish run to the flaking machine,
a mechanical arrangement for distributing the fish evenly upon
heavy wire screens upon which they go to the steam boxes for
cooking. In some factories this flaking is done by hand, but if
the machine is well made and properly run it will do the work
much faster, although it is inevitable, especially when the flsh
are fat and tender, that many are broken. From the steam
boxes the fish are run into a drier where they are kept long
enough to thoroughly dry and, upon emerging from this process,
and sufficient time having ^lapsed for them to cool, they are
taken at once to the packing tables. The packers are girls and
women of all ages who cut off the heads with shears and pack
the fish in cans so rapidly that although they are paid only about
sixteen cents per case of one hundred cans, they earn at times
three or four dollars a day.

The small fish are for the most part packed in oil in small
cans holding four ounces. These are called "quarter oils."
The larger fish are usually packed in mustard sauce in cans
holding eleven ounces. These are called "three-quarter mus-
tards,"

Usually the oil is placed in the can after the fish but the mus-
tard sauce is put into the cans first. After the cans are filled
they are taken to the seaUng machine and when the covers are
in position they are processed in a tank of boiling water from
one and a half to two and a half hours. This process is called
bathing. They are then cooled, cleaned, examined for leaks,
"fats" and "slacks," and finally packed in cases for shipment.

134 Original Communications: Eighth International [vol,

"Fats" are cans which have been carelessly filled too full and,
for that reason might afterwards be mistaken for "swells,"
which have spoiled by fermentation. Such cans are punched
and oil drawn out to reduce the swelUng, after which they are
again sealed and bathed. "Slacks" are those which do not
contain enough and these are punched, heated and oil drawn
in to fill them out.

There are a number of modifications of this general process
in use in various places. A few packers behead all fish before
cooking. A few fish are fried or baked instead of being steamed.
Some of the quarter-oils contain a bay leaf or a clove and there
are a few fish put up in tomato sauce. Several different styles
of ovens and drying machines are in use and there are many
different types of sealing machines, a new one now being per-
fected apparently being a great improvement over any other
•previously made. Whereas all other machines now in use seal
the cans by rolling the edges of the can and cover tightly to-
gether, the new machine seals them hermetically. This machine
also automatically introduces into the cans the proper amount
of oil, fluxes the edges, places the covers and, after tightly
sealing, turns them out at the end ready for the bath at nearly
double the rate of the old machines. An endless belt is arranged
to bring the cans to this machine directly from the packing tables.

There is always great waste of fish during the process. A
twenty per cent loss of the catch is always reckoned upon and
it sometimes runs as high as fifty per cent. When fish break in
large numbers during the process of preparation for the cans
as happens under certain conditions, the waste is large. When
large fish which would ordinarily go as "mustards" are cut to the
size of "quarter oils" the waste is again large for the fish are
sometimes cut in two in the middle in order to make them short
enough for the purpose. This latter practice is resorted to, how-
ever, only when particular orders for "quarter oils" must be
filled regardless of the size of the fish. An attempt to utiUze
some of this waste is being made in some factories by canning
it as "deviled fish," but most of the waste is sold at a dollar
and a half per hogshead to the fertilizer factories where the oil
is pressed out and the pomace used in fertiUzer.

xviii] Congress of Applied Chemistry 135

The two most important questions which have been studied
are the cause of swells, that is, cans swelled out by inside pressure
caused by fermentation, and the breaking of the fish during the
process of preparation for the cans.

By correspondence with packers and by investigation in the
factories it seems evident that the cause of the swells is imper-
fect sealing rather than imperfect or incomplete processing or
bathing. Experiments with four-ounce cans ("quarter oils")
and with eleven ounce cans ("three-quarter mustards") were
made at several factories and it was found that in from twenty-
five to thirty minutes after the cans were immersed in the boiling
bath the temperature had risen at the middle of the can to
100 degrees C. The smaller cans are bathed in the various facto-
ries from one and a quarter to two hours, and the larger cans from
one and a half to three hours, the time varying in the different
factories and with different conditions of the different catches.
This would seem to be long enough to thoroughly sterilize the
contents of the can. That this time is sufiicient for such ster-
ilization is indicated by the fact that in all cases where the
hermetically sealed cans are now put out there is practically no
complaint from swelled cans. This fact, taken in connection
with the other fact that with the ordinary roll sealed cans leaks
often occur and are found even before the goods have left the
factory, would seem to place the blame for the swelled cans
almost entirely upon the sealing machines which do not thoroughly
close the cans. Corroborative evidence is furnished by the fur-
ther fact that practically no living organisms were found on
opening thoroughly sealed cans.

There are at least six different causes contributing to the
breaking of the fish during the process of preparation for the
market.

First. Fish that have lain in a shallow bay or over mud flats
for several days will have softer flesh than those taken from
deeper, cooler water, and are, of course, much more easily
broken.

Second. Fish which are very fat are naturally more tender
than fish which are lean and will break more easily, especially
if they are not properly salted.

136 Original Communications: Eighth International [vol.

Third. The rough, careless handling which many of the fish
receive would break even the firmest fish before they were
finally placed in the cans.

Fourth. If fish are over salted they tend to break transversely
across the side after steaming and drying.

Fifth. When the fish have undigested food in them softening
and breaking commences quicker than otherwise.

Sixth. When the fish are kept too long out of water without
sufficient salt, as is sometimes the case, they naturally begin to
soften and decompose.

It has appeared to some that the breaking of sardines indi-
cates that they are unfit for food. That this is not so is evident
from the above. Of the six reasons given for the breaking only
the two last could be considered as evidence of unfitness for
food, and that the first of these is sufficient evidence of unfitness
is questioned. That those which have burst from the last cause
are unfit for food is, of course, unquestioned and, as their unfit-
ness is at once made evident by the odor, they should be sent to
the fertilizer factory. The breaks which occur in these fish are
somewhat characteristic of the cause of the breaking. For
example, as above stated, over salted fish after steaming and
drying tend to break transversely along one side; fat fish tend
to break along the backbone where the fat is deposited thickly
and the skin is tender; feedy fish, that is fish with undigested
food in them, soften very rapidly and tend to break along the
under side where the flesh is thinnest. Rough and careless
handling increases the tendency to break in all the other cases.
That the breaking along the under side of feedy fish does not
necessarily indicate unfitness for food would appear from the
fact that regardless of thorough salting and careful handling
the breaks begin to appear in about three hours after the fish
are taken out of the water, but fish so breaking give no evidence
whatever of either decomposition or decay, are as sweet and
palatable, and, aside from appearance seem as good as those
not broken.

Feedy fish, above mentioned, are usually full of either one or
the other of two kinds of food known to the fishermen as "shrimp "
and "red feed." This latter is the bane of the sardine industry

xviii] Congress of Applied Chemistry 137

as it has long been known that fish containing it deteriorate
much more rapidly than fish in any other condition. They often
begin to break open before reaching the factory, and, if they
contain much of this material, by the time they are ready for
the can they are broken so badly that a large percentage of the
catch is entirely unfit in appearance for packing. The study
of the subject of breaking involved the questions: "What is
'red feed'? Why does it cause this deterioration?"

This "red feed" we have identified as one of the copepods,
Temora longicornis, a microscopic crustacean of the family
Centropagidce. It is quite abundant in the region of Woods
Hole, Mass., during the winter months and is described by Dr.
William Morton Wheeler in a bulletin of the United States Fish
Commission for 1900. Dr. Wheeler speaks of it as an essentially
boreal form rarely seea in the above locality during the months
of July and August. These are the months in which it is most
abundant along the coast of Maine. Dr. F. H. Moore of the
United States Fish Commission in a report of his investigations
in the herring fisheries of Maine some years ago speaks of this
crustacean as one of the copepods but apparently the species
was not determined. He calls it in his bulletin "red seed" but
this is evidently an error either in printing or in information.

It has loag been recognized by sardine canners that fish con-
taining "red feed" deteriorate much more rapidly than those
containing any other kind of food but why this is so no one to
our knowledge has ever before attempted to determine.

Althougth the sardines were never more abundant on our
coast than during the season just passed, "red feed" was not
particularly troublesome, so that not enough was obtained upon
which to make thorough investigations, but two possible answers
to the second question, the cause of the rapid deterioration,
have suggested themselves. First, that the deterioration is
due to auto-digestion induced by some particular enzyme.
Second, and this seems more likely in the light of present knowl-
edge, that a methylamine is responsible for the trouble. This
compound has been identified in various fish and crustaceans.
It was reported many years ago as being present in the roe of
herring and it is not impossible that in this case "red feed" was

138 Original Communications: Eighth International [vol.

mistaken for roe. It has been reported very recently by Bigelow
and Bacon as being present in considerable amount ia the
shrimps which are canned so extensively in Mississippi. In their
investigation of this problem which was reported in the Journal
of Industrial and Engineering Chemistry for November 1911,
it was noted that this compound affected the hands of the
workmen and had a corrosive effect upon materials which came
in contact with the shrimp containiag it. A parallel case seems
to be found in the sardine industry, for at times when "red
feed" is abundant the hands of the operatives who work upon
the fish are made sore and at times also a strong odor of ammonia
is noted during some of the processes. Either auto-digestion
produced by enzymic action, or the presence of a methylamine,
would probably cause the rapid breaking down of the tissues
and the consequent softening and breaking of the sardines
containing "red feed" and it is hoped soon to further investigate
the question.

In closing it should be stated that the credit for a large part
of the foregoing should be given to A. M. Buswell, Instructor
in Industrial Chemistry in the University of Maine, who acted
as field agent during the summer of 1911, and thanks are also due
Dr. 0. A. Johannsen of the Maine Experiment Station for aid
in identifying the "red feed."

Maine Agricultural Experiment Station,
Orono, Me., U. S. A.

ETUDE CHIMIQUE DES FRUITS DE SORINDEIA

OLEOSA

Par M. Alexandre Hebert
Paris, France

I. — La matiSre premiere de cette 6tude consistait en fruits
s^ch^s au soleil de Sorindeia Oleosa A. Chev. qui nous avaient
6t6 adress^s par M. Augusts Chevalier et qui provenaient d'un
arbre commun au Soudan. Ce sont des fruits k noyau entour6
de pulpe et de la grosseur d'une cerise; ils ont deux usages et
ont 6t6 examines k deux points de vue :

1° — La pulpe ou p6ricarpe du fruit est tihs sucr^e; dans le
pays d'origine, on fait fermenter ces fruits pour en obtenir une
boisson analogue au cidre; 2° I'amande de la graine propreraent
dite, qui forme le noyau du fruit, est tr^s ol6agineuse; on en
extrait de I'huile et on en prepare du savon.

II convenait done de verifier, d'une part, la nature et la pro-
portion du sucre existant dans la pulpe du fruit; d'autre part,
la quantity et les propri^t^s de la mati^re grasse contenue dans
les amandes.

II.— Pour effectuer I'^tude chimique de ces fruits aux points
de vue qui nous int6ressaient, nous avons commencd par s^parer
les pulpes et les noyaux. A cet effet, 500 grammes de ces fruits
s^ch^s ont 6t6 mis en contact avec une quantity d'eau froide
suflBsante pour les recouvrir, aprSs 24 heures de s^jour, ils
s'^taient gonfl^s et ^taient d'une consistance telle qu'ils pou-
vaient ^tre malaxes dans I'eau sans risquer d'^craser les noyaux.
Ceux-ci, s6par6s ainsi des pulpes, ont 6t6 dess^ch^s k I'air et
mis de c6td pour un examen ult^rieur. On les a trouv6s en pro-
portion de 40 pour 100 des fruits sees accusant ainsi 60 pour 100
de pulpes.

liCs pulpes gonfl^s ont 6t€ 6puis6es a trois reprises par I'eau
froide pour dissoudre toutes les mati^res solables et notamment
les sucres qui s'y trouvaient. Finalement le r^sidu a €t4 press6

139

140 Original Communications: Eighth International [vol.

et le liquide provenant de ce pressurage a et6 joint aux liqueurs
d'^puisement. Celles-ci ont 6t6 d6f^qu6es par le sous-ace-
tate de plomb et le liquide filtr4 a 6t4 d^barrass^ de I'exc^s de
plomb par I'hydrog^De sulfur^. La solution incolore ainsi
obtenue a ^t^ concentr^e dans le vide au bain-marie k tr^s basse
temp^rat.ire jusqa'a consistance sirupeuse, puis abandonn^e k
elle-meme. Elle a refas6 de cristalliser, malgr^ to us les subter-
fuges habituels employes dans ce but: concentrations diverses,
reprises par I'alcool traitement au noir animal, etc. Le sirop
r^duisait ^nergiquement la liqueur de Fehling, donnait avec
I'ac^tate de ph^nyl-hydrajine une osazone cristallis^e en aiguilles
groupies en forme d'^ventail, fusibles k 200° et correspondant
aux propri^t^s de la ph^nyl-glucosazone, deviait enfin k gauche
le plan de polarisation de la lumi^re, mais cette deviation cor-
respondait k une quantity de sucre r^ducteur bien plus faible que
celle indiqu^e par le titrage k la liqueur de Fehling. Somme
toute, ces caractSres r^pondaient au sucre interverti.

D'autre part, on a trouv6 dans une quantite donn^e des
fruits sees, ^puis^s par I'eau froide comme nous I'avons indiqu^,
et par titrage k la liqueur de Fehling, une proportion de 22 pour
100 de sucres r^ducteurs et une quantity nulle de sucres non
r^ducteurs. Si nous admettons dans ces fruits, k I'^tat frais,
une teneur en eau igale k 90 ou 95 pour 100, teneur qu'on retrouve
g^n^ralment dans les fruits de ce genre, la proportion de sucres
r^ducteurs correspondraic k 1:10 ou 2:20 pour 100 des m^mes
fruits k retat frais.

II r^sulterait de ces i.xp^riences que les mati^res sucr^es des
fruits de Sorindeia Oleosa A Chev. seraient constitutes par du
sucre interverti, melange de glucose et de l^vulose, ce qui justi-
fierait leur emploi indigene pour la preparation d'une boisson
plus ou moins alcoolique, et du genre du cidre, mais qui, en tons
cas, ne peut certainment etre que trSs peu riche en alcool.

III. — Les noyaux, obtenus comme nous I'avons dit, et qui
constituaient 40 pour 100 des fruits sees, renferment 24 pour
100 de ces memes fruits sees en amandes. Celles-ci, aprSs broyage
et extraction k la benzine, lui abandonment une matiSre grasse
dont la proportion atteient 25 pour 100 des fruits sees.

xviii] Congress of Applied Chemistry 141

La matifere grasse obtenue est solide k la temperature ordi-
naire, de couleur brun^tre et pr^sente les constantes suivantes:

Density ^17° 0.889

Point de fusion 16-17°

Point de congelation 12-13°

Indice d'acidite 4.90

Indice de saponification 185.00

Indice de Reichert 7 . 92

Indice d'Hehner 91 .75

Indice d'iode 132.00

La graisse de Sorindeia Oleosa A. Chev., saponifiee par la
soude alcoolique et acidifi^e, fournit 92 pour 100 environ d'acides
gras, jaunitres, solides k la temperature ordinaire, fusibles k
39-40°. La separation des acides gras satures et incomplets
effectuee par I'epuisement k I'ether des sels de plomb, a donne
24 pour 100 d'acides incomplets, liquides, de couleur jaune
brunitre, et 76 pour 100 d'acides satures, solides, colores en
jaune brun, fondant k 44-45°. Ce point de fusion assez bas
indique I'existence, dans la graisse etudiee, d'acides gras rela-
tivement inferieurs. L'usage de la graisse de Sorindeia Oleosa
A. Chev. pour la preparation du savon se comprend ainsi par-
faitement, cette substance grasse d'une part, ne paraissant pas
comestible, et d' autre part, donnant des acides gras k point de
fusion trop bas pour servir k la fabrication de bougies ou m^me
de chandelles.

SUR LA COMPOSITION DE DIVERS PRODUITS,
GRAINES OU TUBERCULES AMYLACES OU FECUL-
ENTS DE L'AFRIQUE OCCIDENTALE FRANgAISE

Pab M. Alexandre Hubert
Paris, France

Au cours de sa demifere mission en Afrique occidentale fran-
gaise, M. Aug. Chevalier a rapports un certain nombre de pro-
duits, graines ou tuberciiles, de nature amylac^e ou f^culeate,
qu'il nous a remis pour en determiner la composition chimique,
en appr^cier la valeur nutritive et en fixer I'emploi industriel
possible. Ce sont ces diverses recherches que nous r&umons ici.

GRAINES. — Mais blanc du Dahomey. — Ce mais nous a 6t6
remis sous forme d'^pis dont une certaine quantity ^taient mal-
heureusement charangonn^s; nous avons pu cependant en
trouver quelques-uns intacts sur lesquels nous avons efifectu^
I'analyse. Nous avons s6par6 dans les 6pis les glumes et glu-
melles, les rachis et les graines dont nous avons determine la com-
position. Nos dosages nous ont conduit aux r^sultats suivants:

SicMs a I'air S^cMs d 110°

Poids moyen d'un 6pi entier 120 gr. 110 gr.

Decomposable en glumes et glumelles. ...16.6 16.6

graines 88 . 3 78 . 3

rachis 15.0 15.0

Analyse de la graine s^chee et moulue

Humidite restant 1 . 63%

MatiSres min^rales . . . . 1.96 dont 0.49 solubles dans I'eau.

Matififies grasses 3 . 70

Mati^res azot^es 11 . 55 dont 0 . 98 solubles dans I'eau.

Sucres r^ducteurs . . . . 0 . 36
Sucres non r^ducteurs . 0 . 95
Gommes, tannins,
acides v6g6taux. ... 0.24

Amidon 76.30

Cellulose 1.36

Vaaculose 1 . 90

Total 99.95

143

144 Original Communications: Eighth International [vol.

Cette graine peut done ^tre comparfe, au point de vue de sa
valeur, k nos produits indigenes. EUe est d'autant plus int^res-
sante qu'il s'en exporte d'Afrique des quantity importantes
dont I'introduction pourrait rendre service a diverses industries.

Voandzeia Poissonni A. Chev. — Ces grain es qui proviennent
d'Ouaga dougou (Mossi), pr^sentaient la composition ci-dessous.

Humidity 10.38

Matieres minerales 4 . 34

Mati^res grasses 1.91

MatiSres azot^es 21.41

Sucres r^ducteurs traces

Sucres non r^ducteurs 0.41

Amidon 48 . 77

Cellulose 12.74

, Total 99.96

Cette graine, riche en matieres azot^es, renferme moins
d'amidon que les graines amylac6es de nos pays;, elle peut
n^anmoins etre employee au point de vue alimentaire au moins
dans les contr^es d'origine.

TUBERCULES. — Ignames. — Ces tubercules qui nous ont
6te envoyfe k I'^tat dess^ch^, provenaient de la cote d'lvoire;
lis ont donn6 k I'analyse les r^sultats suivants :

Humidity 13.80

Matieres minerales 2 . 40

Matieres grasses 0 . 40

Matieres azot^es 5 . 75

Sucres r^ducteurs 1 . 00

Sucres non r^ducteurs 1 . 00

Amidon 73 . 80

Cellulose 1 . 25

Vasculose 0 . 60

Total 100.00

Ces tubercules sont assez comparables comme composition
k la pomme de terre. Ceux qui ont 6t6 exp6di6s en Europe ont

XVIII

Congress of Applied Chemistry

145

6t6 trouv^s de valeur au moins ^gale au manioc sec. Le com-
merce des ignames africains pourrait done prendre de I'exten-
sion comme produit alimentaire sous une forme quelconque.

DIEGEMTENGUERE (Vulg. Mossi).— Les tubercules de
cette plante qui nous ont 6t6 remis provenaient d'Ouagadougou
dans le Soudan frangais. Leur poids avait it^ d6termin6 k
I'^tat frais ce qui nous a permis de fixer leur composition exacte
k r6tat frais et k I'^tat sec:

Eau

Matidres min^rales . . .

Mati^res grasses

Matidres azotdes

Sucres rfiducteurs ....
Sucres non r6ducteiirs

Amidon

Cellulose

99.94

Etat frais

Etat sec

67.90%

0.00%

2.02

4.80

0.21

0.50

4.47

10.62

N6ant

2.69

6.40

28.80

68.40

3.85

9.15

99.87

Cette composition ratifie parfaitement I'emploi de cette plante
qui est cultiv^e au Mossi, dans la boucle du Niger, pour ses
tubercules alimentaires.

MOELLE d'ENCEPHALARTOS BARTER!.— Ce produit
est extrait d'une plante de la famille des Cycadac^es, dont la
tige est pourvue d'une moelle abondante qui poss^de la compo-
sition suivante :

Humidity 12.80

Mati^res min^rales 2 . 80

Mati^res grasses 0. 60

Mati^res azot^es 6 . 43

Sucres r^ducteurs 10 . 00

Sucres non r^ducteurs 1 . 10

Amidon 60 . 52

Cellulose 4.25

Vasculose 1-50

Total 100.00

146 Original Communications: Eighth International [vol.

Cette moelle est, comme on le voit, riche surtout en hydrates
de carbone : sucres et anaidon ; cette richesse justifie parfaitement
I'emploi indigene que I'on fait de cette moelle en fabriquant
une sorte de pain avec la f^cule qu'on en extrait.

(Abstract)

ON THE TASTE OF THE SALT OF GLUTAMIC ACID

KiKUNAE IkEDA

College of Science, Imperial University of Tokyo, Tokyo, Japan

The glutamates having the general formula C6H8NO4M' are
mostly soluble in water and all of them have a very distinct
peculiar taste, the quality of which differs from all other well
defined taste qualities hitherto known. Numerous food materials
present this taste, but so much overshadowed by others, that no
clear conception of this quality has hitherto been formed, although
it contributes largely to the flavor. For this taste quality the
name "glutamic taste" is proposed.

This taste is then demonstrated to be that of the monovalent
glutamate ion C^HsNO^. For this purpose the threshold value of
the taste has been carefully measured for the salts of sodium,
potassium, magnesium, calcium and barium. The value has
been found to be ^^^u normal for all the five salts. The taste-
imparting power of the glutamates is very great.

The author was led to the discovery of the taste of glutamates
by his investigation on the constituents of a certain sea-weed,
which is used in Japan as a flavoring. He isolated glutamic acid
from it and found that it is the salts of this acid that give the weed
its peculiar flavor.

There are numerous flavoring substances which give glutamic
taste, and among them meat-extract and allied preparations.
But from obvious reasons a pure glutamate is much to be pre-
ferred over them. Of all the glutamates of non-poisonous
metallic radicals the sodium salt is the most suitable. Within the
last three years the manufacture of this salt has arisen in Japan
and it is now rapidly becoming an article of general consumption.

There is hardly any doubt that the glutamate will come to be
manufactured in a large scale in Europe and America. As the
raw material for the manufacture is the hydrolytic products of
proteins, there is a prospect that the chemical industry of these
products will be greatly developed, bringing in its train numerous
problems of great interest.

147

PROGRESS REPORT OF NUTRITION INVESTIGATIONS
IN THE UNITED STATES

By C. F. Langworthy, Ph.D.
Office of Experiment Stations, Department of Agriculture

Introduction

For many years, continuous progress has been made in the
United States in the study of various questions concerned with
human nutrition. In this summary, the attempt has been made
to bring together articles on this subject which have appeared in
the United States, since the 7th International Congress of
Applied Chemistry, thus supplementing a paper of similar
scope presented at the 7th Congress.

A survey of the literature under consideration shows that a
considerable part of it represents work carried on under govern-
mental or institutional auspices, a considerable part represent-
ing the work of the United States Department of Agriculture and
other branches of the general Government and the agricultural
experiment stations. University laboratories and the labora-
tories of endowed institutions are also large contributors as are
also state boards of health.

In general, it may be said that judging by the amount of work
which is published annually, interest in the experimental study
of human nutrition is growing very rapidly. The fact is rec-
ognized that the record of work here presented is by no means
complete but it is believed that it is sufficiently extended to show
the character and scope of the work which is being done.

For convenience the material has been arranged under the
following heads: Studies of Food and Food Products; Special
Studies of Ash, Protein, and Other Food Constituents; Cooking
in Its Relation to Nutritive Value; Canning, Preserving, Han-
dhng and Storage; Dietary Studies and Dietetics; Digestion;
Metabolism; Respiration Calorimeters, Bomb Calorimeters, and
Experiments with Them; Foods and Their Relation to Problem
of Hygiene; and Cost of Living and Other Statistical Data.

149

150 Original Communications: Eighth International [vol.

Studies of Food and Food Products.

As is usuaEy the case, considerable attention has been given
to the proximate composition of food products and to the effect
of various processes of manufacture or handling upon nutritive
value and quality. Many hundreds of proximate analyses, more
or less complete in character, have accumulated during the
periods under consideration, in connection with inspection work
under national and state pure food laws and as a part of other
work undertaken for some special purpose aside from analysis.

Many milling and baking tests with different varieties of wheat
have been reported, this question being one which is of great
interest particularly in wheat-growing regions. E. F. Ladd and
Emily E. May (North Dakota Sta. Spec. Buls. 19, pp. 105-114;
24, pp. 179-194, fig. 1) have carried on extensive work of this
sort with durum wheat flour. Their studies showed that more
power was required to grind durum than Fife or Bluestem but
the yield was as large and the bread made equal to that produced
from other flours, though not quite so white in color. It was
found to hold moisture better than that from commercial flours.
A study of milling and baking of durum wheat flour was reported
by L. M. Thomas of the North Dakota Experiment Station.

The effect of climatic conditions on the composition of durum
wheat has been discussed on the basis of a number of analytical
and other studies carried on bj- the Department of Agriculture,
by J. A. LeClerc (U. S. Dept. Agr. Year Book, 1906, pp. 199-212,
pis. 2.) Several hundred analyses of spring and winter wheat of
different varieties grown in different States showed an average
protein content of 12.2 per cent as compared with 14.7 per cent
for over 100 samples of durum wheat analyzed by the author.

E. F. Ladd and C. H. Bailey (North Dakota Sta. Buls. 89, pp.
14-80; 93, pp. 204-253, dgms. 5) have reported an extended
study of the milling quality of wheats of different varieties and
crops.

Similar tests of wheat of different kinds and crops grown in
different localities have also been made by R. W. Thatcher
(Washington Sta. Bui. 84, pp. 48, figs. 3) and L. R. Waldron
(North Dakota Sta., Rpt. Dickinson Substa. 1910, pp. 43, 44),

xviii] Congress of Applied Chemistry 151

and with California wheats by G. W. Shaw and A. J. Gaumnitz
(California Sta. Bui. 212, pp. 315-394, figs. 18, dgms. 3), and
with a variety of wheats by F. D. Gardner (Roller Mill, 28 (1909),
No. 5, pp. 201-204).

It is interesting to note the discussion of the future wheat
supply of the United States by M. A. Carleton (U. S. Dept. Agr.
Yearbook 1909, pp. 259-272, figs. 2), which is based on a digest
of statistical data.

With respect to the effect of soaking and germination of wheat
on the distribution and yield of milling products, the quality of
flour, and bread-making properties, G. A. Olson (Amer. Food
Jour., 6 (1911), No. 4, pp. 36-39, figs. 4) found that water-soaked
wheat is not necessarily spoiled and can be used for milling pur-
poses, providing it has been thoroughly cleaned and dried. Using
small quantities of germinated wheat flour with other flour in-
creased the volume of the loaf, according to the author, without
impairing its texture. Each particular flour required a different
amount of germinated flour to produce the best results. Too
large an amount of diastatic flour is less beneficial than none.

Analyses of a number of sorts of gluten flour manufactured
in the United States and of foreign diabetic products were re-
ported in comparison with wheat flour by D. W. Fetterolf (Univ.
Penn. Med. Bui., 22 (1909), No. 7, pp. 217-222).

From an experimental study of the starch grain, H. Kraemer
(Amer. Jour. Pharm., 79 (1907), pp. 217-229, pi. 1, figs. 3) con-
cludes that "the starch grain consists of colloidal and crystal-
loidal substances, these being arranged for the most part in dis-
tinct and separate lamellae, that is, at the point of origin of growth
and in the alternate lamellae the colloidal substance preponder-
ates, associated with the crystalloid cellulose; whereas in the
other layers the crystalloidal substance, consisting for the most
part of granulose, occurs in greater proportion."

Several studies of cane sugar and maple sugar have been car-
ried on.

C. A. Browne, Jr., and R. E. Blouin (Louisiana Stas. Bui. 91,
pp. 103) have summarized a large amount of data collected dur-
ing recent years by the Louisiana Sugar Experiment Station,
which have to do with the composition of the stalk, sesd, root.

152 Original Communications: Eighth International [vol.

and leaves of sugar cane and of the plant ash. The work as a
whole is an exhaustive study of the chemical composition of sugar
cane, with reference to its use for sugar making, and of the phy-
siology of the growth and ripening of the cane. Experimental
work on sugar making is also reported.

From a study of the question of the influence of micro-organ-
isms upon the quality of maple sirup, H. A. Edson (Abs. in
Science, n. ser., 31 (1910), No. 791, p. 308) was able to show by
isolation and inoculation experiments that to certain groups of
micro-organisms is ascribable the abnormal type of sap of the
late runs characterized by green, red, milky, and stringy appear-
ance.

A. H. Bryan (U. S. Dept. Agr., Bur. Chem. Bui. 134, pp. 110,
pi. 1, figs. 4, map 1), in connection with a study of maple sap
sirup, repoits analyses of 481 samples of sirup of known purity
collected in maple-producing States in the United States and in
Canada, the data being gathered as a basis for comparing and
grading maple sirups.

Considering the 395 samples from the United States, the aver-
age moisture content was 34.19, sucrose 62.64, invert sugar 1.49,
ash 0.66, and undetermined material 1.02 per cent. The polar-
ization values were: Direct, at 20°C.,-f60.93; and invert, at
20°C.,— 22.16. The average values for the 86 Canadian sam-
ples were: Moisture content 34.34, sucrose 62.24, invert sugar
1.41, ash 0.62, and undetermined material 1.59 per cent. The
polarization values were: Direct, at 20°C.,-|- 59.33; and invert,
at20''C.,-23.17.

The results of a special study of the constituents of maple-
sirup ash are also reported. The average results for 100 sam-
ples from different States showed that the ash contained 38.07
per cent potash, 21.88 per cent lime, 5.39 per cent phosphoric
acid, and 1.59 per cent sulphates.

Considering the samples from both the United States and
Canada, the average basic lead value of 2.70, calculated to dry
substance, and the average neutral lead number was 0.79. The
average malic acid value determined by the modified calcium
chloride method was 0.84, and by the calcium acetate method,
1.01.

xviii] Congress of Applied Chemistry 153

Factors which influence the character of the sap and the sirup
and related questions are discussed.

Experimental work carried on in an attempt to isolate flavor-
ing substances present in maple sap is described by A. P. Sy
(Jour. Franklin Inst., 166 (1908), pp. 249-280); Abs. in Chem.
Abs., 2 (1908), No. 24, p. 3376), in a publication dealing with
history, manufacture, and analysis of maple products, and work
reported on the analysis of maple products.

Housekeepers and manufacturers of food products often ex-
press the opinion that there is a difference in the culinary quality
of cane sugar and beet sugar. The matter was studied by G. W.
Shaw (California Sta. Circ. 33, p. 4), of the California Experi-
ment Station. The sugar is being used for sirup making, for
canning fruit, and for jelly making. The beet sugar produced
more froth in making sirup, but investigation led to the conclu-
sion that this was due to the finer granulation of the beet sugar,
which caused more air to become entangled during the starting
than was the case with cane sugar. No differences were ob-
served in the keeping quality of canned goods or the jelly made
with the two sorts of sugar from his experimental data and
other evidence the author concludes that under commercial and
household conditions, beet sugar and cane sugar give equally
satisfactory results for these uses.

An exhaustive study was made of the composition of Ameri-
can honeys by C. A. Browne (U. S. Dept. Agr., Bur. Chem. Bui.
110, pp. 1-69, 89-93, pi. 1, fig. 1), of the Bureau of Chemistry,
and a microscopical study of honey pollen by W. J. Young (U. S.
Dept. Agr., Bur. Chem. Bui. 110, pp. 70-88, pis. 5).

A number of studies of meat, eggs, cheese, and other animal
foods have appeared.

The glycogen content of beef flesh and the factors which in-
fluence it were studied, in animals recently slaughtered, by P. F.
Trowbridge and C. K. Francis (Jour. Indus, and Engin. Chem.,
2 (1910), No. 1, pp. 21-24). The length of time which elapses
after feeding before the animal is slaughtered, the authors con-
sider an important factor in determining the amount of glycogen
which remains stored in the organs and muscles. Their results
indicate that there is a rapid enzymatic hydrolysis of glycogen

154 Original Communications: Eighth International [vol.

in flesh under many conditions, but that at 10°C. or lower, it
did not take place appreciably.

In connection with an extended study of market classes and
grades of meat, L. D. Hall (Illinois Sta. Bui. 147, pp. 147-290,
figs. 75; Abstract, pp. 15, figs. 4) has described and illustrated
by diagrams or figures the standard grades of beef, veal, mutton,
and pork as they are found in the Chicago wholesale trade. Tech-
nical terms are defined. The bulletin as a whole furnishes a
large amount of data on the subject which is of importance in
discussing meat in relation to dietetics as well as for other pur-
poses.

Some data of a similar character have been published by P. F.
Trowbridge (Missouri Bd. Agr. Mo. Bui., 9 (1911), No. 2, pp.
69-78).

W. D. Richardson (Jour. Amer. Chem. Soc. 29 (1907), No. 12,
pp. 1757-1767) reports the results of the examination of a large
number of samples of animal and vegetable foods with a view to
securing data regarding the occurrence of nitrates in vegetable
foods, cured meats, and elsewhere. He concludes that nitrates
are quite generally distributed.

F. C. Cook (U. S. Dept. Agr., Bur. Chem. Ciro. 62, pp. 7), of
the Bureau of Chemistry of the Department of Agriculture, has
reported a large number of analyses of beef extracts and yeast
extiacts of known origin. According to the author, "the yeast
extracts contain approximately 1 per cent ether-soluble mate-
rial and the beef extracts larger amounts. Cholesterol was not
found in the ether extracts, and sarcolactic acid only in the yeast
extracts.

"The phosphorus of beef is largely water-soluble, consequently
a considerable percentage of the ash of beef extracts is composed
of this constituent. Approximately one half of the sulphur of
beef is water-soluble. Yeast extracts derived from yeast rich
in phosphorus also contain a large amount in the ash. The
total amount present is larger than the ash content, showing that
some phosphoric acid is volatilized on ashing. The organic
phosphorus determined by the Siegfried-Singewald method gives
approximately the 1:10 ratio compared with the total as sug-
gested by those authors.

xviii] Congress of Applied Chemistry 155

"The total nitrogen of the beef extracts on the water-free and
fat-free basis averages 11.82 per cent, that of the yeast extracts
averages 7.44 per cent. The amino nitrogen figures for the beef
preparations are nearly double those of the yeast extracts.

"Although the water-soluble nitrogen of beef, which consti-
tutes 25 per cent of the total nitrogen, consists of approximately
two thirds and one third amino nitrogen, the samples of beef
extracts analyzed average 72 per cent of amino nitrogen and 28
per cent of protein nitrogen.

"The general appearance and odor of the two varieties of ex-
tracts are very similar. As a food both are extremely limited in
value. The beef extracts contain more nitrogenous extractives
than the yeasc preparations, otherwise their general composition
is much the same."

A large number of analyses of samples of meat extract, meat
juices, yeast extracts, and similar goods are reported and dis-
cussed by W. D. Bigelow and F. C. Cook (U.S. Dept. Agr., Bur.
Chem. Bui. 114, pp. 7-56), the methods followed being described.

Meat extracts, yeast extracts, and similar goods were also
tudised by J. P. Street, et. al. (Connecticut State Sta. Rpt. 1907-8,
pt. 9, pp. 573-716).

On the basis of numerous tests, I. A. Field (U.S. Dept. Com.
and Labor, Bur. Fisheries Bui., 28 (1908), pt. 1, pp. 243-257;
Doc. 655, 1910, pp. 243-257) reaches the conclusion that the
common sea mussel {Mytilus edulis) is nutritious, palatable,
and easily digested. From tests of culinary qualities, made
under a variety of conditions, of the smooth and horned dogfish,
he concludes further that the flesh of these fishes is cheap, pala-
table, nutritious, and easily preserved, and he believes further
that it is as digestible as that of other fishes.

In a paper on unutilized fishes and their relation to the fishing
industries, I. A. Field (U. S. Dept. Com. and Labor, Bur. Fish-
eries Doc. 622, pp. 50, pi. 1) discusses methods of proficably using
dogfish of different sorts, sand shark, toad-fish, etc., summarizes
data regarding the use of fresh, canned and dried dogfish, and
gives some results of tests of its culinary quality, which he be-
lieves indicate that such dogfish flesh is both palatable and
wholesome.

156 Original Communications: Eighth International [vol.

The uniformity with which copper was found in oysters ex-
amined by J. T. Willard (Jour. Amer. Chem. Soc, 30 (1908),
No. 5, pp. 902-904) led him to conclude that it is to be regarded
as a normal constituent.

J. T. Willard and R. H. Shaw (Kansas Sta. Bui. 159, pp. 143-
177) analysed all the eggs laid in 6 weeks by 4 lots of pure-bred
chickens. On an average the thickness of the shells was 0.0139
in. In addition to usual determinations, they report data re-
garding the percentage of phosphoric acid, the ash in the yolk,
and the ratio of phosphoric acid to ash. The average amount
of ash was 1.57 per cent and of phosphoric acid 1.43 per cent,
the ratio of phosphoric acid to ash being 1:1.09.

"It is evident that the ash consists almost entirely of phos-
phoric acid. This is doubtless produced almost entirely, if not
altogether, from the lecithin of the egg yolk."

Mary E. Pennington (Jour. Biol. Chem., 7 (1910), No. 2, pp.
109-132) has reported the results of an extended chemical and
bacteriological study of fresh eggs, which was reported at the
London Congress of Applied Chemistry, in June, 1909, and later
published in full.

L. L. Van Slyke and A. W. Bosworth (New York State Sta.
Tech. Bui. 4, pp. 1-16, 17-22) at the New York State Station
have studied some of the early chemical changes which take place
in the proteids and in the calcium and phosphoric acid compounds
of Cheddar cheese, and also the acidity of the water extract of
Cheddar cheese.

A. W. Bosworth (New York State Sta. Tech. Bui. 5, pp. 23-39)
has also reported the results of chemical studies of Camembert
cheese.

The manufacture of a food product called buttermilk is de-
scribed by J. L. Sammis (Wisconsin Sta. Bui. 211, pp. 3-17, figs.
7), in a bulletin of the Wisconsin Experiment Station and some
data given regarding its fat content, keeping qualities, etc.

An experimental study of the production of a dairy product
called "whey butter" has been reported by C. F. Doane (U. S.
Dept. Agr., Bur. Anim. Indus. Circ. 161, pp. 7).

G. A. Olsen (Jour. Biol. Chem., 5 (1908), No. 2-3, pp. 261-281)
reports data regarding a proteid found in milk, cream, and but-

xviii] Congress of Applied Chemistry 157

ter which he considers new. The chemical and physical char-
acter of this proteid are described.

The majority of investigations with fruits and nuts carried on
in the United States have had to do with the methods of culti-
vation, transportation, and shipment rather than with compo-
sition, food value, and use in the home.

Cactus fruits, particularly tuna or the fruit of the prickly pear,
which is used in southwestern United States and to a greater
extent in Mexico as a foodstuff, were studied with respect to its
composition and nutritive value, by R. F. Hare and D. Griffiths
(New Mexico Sta. Bui. 64, pp. 88, pis. 7, figs. 2).

In the study of the tuna as food for man, by D. Griffiths and
R. F. Hare (U. S. Dept. Agr., Bur. Plant Indus. Bui. 116, pp.
73, pis. 6), information is given regarding the use of the fruit for
jelly making and preserves as well as for other purposes.

Italian lemons and their by-products and methods of produc-
ing lemon oil and citric acid commercially are discussed in a sum-
mary of data by E. M. Chace (U. S. Dept. Agr., Bur. Plant Indus.
Bui. 160, pp. 35-50, pis. 3, figs. 2).

In connection with a summary of data on the dietetic value of
fruit, W. R. Lazenby (Trans. Mass. Hort. Soc, 1910, pt. 1, pp.
89-97) reports data regarding the water content of well-devel-
oped and undeveloped specimens. Less than 80 per cent
water was found in undeveloped strawberries, peaches, and ap-
ples, as compared with 90 per cent in fine but not overgrown
specimens. It is further stated that 92 per cent of water was
found in fine large peaches, in comparison with 84 per cent in
small peaches of the same variety.

Data were also recorded regarding the percentage of shell or
waste, and edible portion in nuts, and similar factors. According
to the author, there is a loss of nearly 2 per cent of the total weight
of kernels in milling or cracking some of the larger sorts of nuts.

Various topics concerned with the composition, nutritive value,
and use of fruit as food have been discussed in a summary pre-
pared by C. F. Langworthy (U. S. Dept. Agr., Farmers' Bui.
293, pp. 38, fig. 1).

The occurrence of sucrose in grapes was studied by W. B.
Alwood and his associates (Jour. Indus, and Engin. Chem., 2

158 Original Communications: Eighth International [vol.

(1910), No. 11, pp. 481, 482) with a number of varieties. The
quantities found in the juice of 3 well-known varieties ranged from
4.49 and 5.66 gm. per 100 cc. of juice. In the juice of a new
seedling it was considerably larger.

In a later report, W. B. Alwood (U. S. Dept. Agr., Bur. Chem.
Bui. 140, pp. 24) states that he and his co-workers have examined
practically all the wine and table grapes grown in eastern United
States, and with the exception of the varieties mentioned (Hayes,
Pocklington, and Worden and a seedling), they did not find
sucrose in appreciable quantities. Extended studies were also
made of varieties grown in other regions and the variations in
sugar and acid content studied during ripening. In Catawba
grapes the sugar more than doubled after the berries began to
color, while the acid was only about half as great. Similar data
are reported for many other varieties.

W. P. Kelly (Jour. Indus, and Engin. Chem., 3 (1911), No. 6,
pp. 403-405) has studied the composition of Hawaiian pineap-
ples and found them to vary considerably, the sugar content
ranging from 9.15 to 15.23 per cent, and the acidity from 0.22 to
1.16 per cent, and increasing generally as the sugar increased.
On the whole, Hawaiian pineapples show much the same average
composition as thos"; grown elsewhere.

"Green pineapples contain less acidity that the ripe fruit and
also a small percentage of fiber, reducing sugar, and sucrose.
Dextrin and starch do not occur in important quantities in pine-
apples at any stage. The reducing sugars and sucrose stand in
inverse ratio to that of the ripe fruit. In the ripening of pine-
apples gathered green, the most important chemical change chat
takes place is the conversion of reducing sugars into sucrose, but
the total sugar content appears not to be increased. . . .

"During the normal ripening of the pineapple, a rapid accu-
mulation of sugais and a slight increase in acidity take place.
When the fruit becomes approximately half ripe, it contains at
least three-fourths of its maximum sugars."

Bread, milk, vegetables, bananas, and rhubarb were included
by H. Ackroyd, (Bio-Chem. Jour., 5 (1911), No. 8-9, pp. 400-
406) in a study of the presence of allantoin in certain foods.

His general conclusions are that "the whole quantity of allan-

xviii] Congress of Applied Chemistry 159

toin excreted by man on a milk and vegetable diet may be derived
directly from the food. Milk, white bread, French beans, green
peas, all contain small quantities of allantoin, while none could
be isolated from eggs, bananas, or rhubarb."

The food value of nuts and the various ways in which they may
be used in the diet have been discussed by M. E. Jaffa (U. S.
Dept. Agr. Yearbook 1906, pp. 295-312, pi. 1, fig. 1; Farmers'
Bui. 332, pp. 28, fig. 1), in a bulletin published in connection
with the nutrition investigations of the Office of Experiment
Stations.

In connection with a study of pecan culture, the marketing of
pecans, and related questions by W. N. Hutt (Bui. N. C. Dept.
Agr., 30 (1909), No. 9, pp. 50, figs. 25), the use of pecans as food
is considered, and instructions given for cracking these nuts par-
ticularly for commercial purposes.

The care and marketing of vegetables have been more often
studied than their composition and nutritive value.

Canned peas and beans of different grades were analyzed by
W. L. Dubois (U. S. Dept. Agr., Bur. Chem. Circ. 54, pp. 9),
in connection with commercial canning, and particularly with
reference to the use of soaked peas and beans in place of the
fresh vegetables. In general, the soaked peas had a higher
water and starch comeni, and a somewhat higher specific
gravity than the fiesh canned peas. The author is of the
opinion that such determinations may prove useful in connec-
tion with physical examinations in judging of the character of
such canned goods.

The crude fiber and the crude search content of the soaked
were higher than in the case of the fresh canned beans, though
the differences were less pronounced when the results were re-
duced to a dry matter basis.

The recorded data furnished some information regarding the
changes which take place during the growth and ripening of peas.

"As the pea matures the ash decreases, the starch increases, and
the crude fiber decreases as a rule, while the conclusions to be
drawn from the determinations of nitrogen and ether extract
are less decisive. In the peas from one locality the amount of
nitrogen decreases as the pea matured, whereas in the same vari-

160 Original Communications: Eighth International [vol.

ety from another locality this variation was not so apparent.
Similar changes in composition appear in the canned vegetables.
The analyses seem to indicate that during the process of canning
the peas take up from 2 to 10 per cent of water. It is difficult
from these results to draw any conclusions as to the changes
taking place during processing. The principal value of the work
. . . is to afford data for the comparison of commercial
grades."

Marine algse are important articles of diet of native Hawaiians.
In connection with the work of the Hawaii Experiment Stations,
Minnie Reed (Hawaii Sta. Rpt. 1906, pp. 61-88, pis. 4) studied
the economic importance and food value of a large number of
these marine algae, reporting cooking tests in addition to analyti-
cal work and studies of the value of seaweed mucilage, gelatin,
etc.

A digest of data on insoluble carbohydrates, particularly those
of marine algse, and a summary of digestion experiments car-
ried on in the author's laboratory with such foods in comparison
with raw ItaUan chestnuts, have been briefly reported by L. B.
Mendel (Zentbl. Gesam. Physiol, u. Path. Stoffwechsels, n. ser.,
3 (1908), No. 17, 641-654).

The character and nutritive value of carbohydrates of lichens,
algse, and related substances, particularly marine algae, as studied
by Mary D. Swartz (Proc. Amer. Soc. Biol. Chem., 1 (1910),
No. 5, pp. 257, 258; Trans. Conn. Acad. Arts and Sci., 16 (1911),
pp. 247-382), the hemicelluloses from 10 species of algse were
found to contain pentosans and galactans. The pentosans, with
one exception, were largely found insoluble in cold water, while
the gelactans were soluble and characterized by their gelatinous
nature. Small quantities of soluble pentosans were associated
with them in every case.

The resistance to bacterial action was studied, and digesti-
bility was studied in vitro, and in other ways.

They were found to be very resistant to the action of animal
and vegetable enzyms. Experiments showed that galactans
were not affected by the ordinary aerobic bacteria of the
alimentary tract, or by mixtures of soil and fecal aerobes,
of soil and fecal anaerobes, or of powerful putrefactive organ-

xvni] Congress of Applied Chemistry 161

isms such as Bacillus anthracis ^mptomatid and B. maliqni
aedematis. Pentosans, mannans, and levulans were found to be
gradually decomposed by soil and fecal bacteria and by putre-
factive anaerobes, sometimes with the formation of reducing
substances.

" When introduced parenterally, either subcutaneously or
intravenously, they are not retained or altered by che organ-
isms, but are gradually excreted in the urine. Feeding experi-
ments on dogs and human subjects show that those hemicelluloses
most readily attached by bacteria disappear most completely
from the alimentary tract. Galactans, which are unaffected to
any appreciable extent, are excreted in amounts averaging 75
per cent; pentosans and mannans, hydrolyzed by bacteria,
disappear almost entirely during the processes of digestion.

" The experiments give little justification for considering
these carbohydrates as typical nutrients for man."

A popular digest of data regarding the composition, food
value, digestibility, and place in the diet of potatoes and other
root crops used as food is prepared by C. F. Langworthy (U. S.
Dept. Agr., Farmers' Bui. 295, pp. 45, figs. 4).

Proprietary foods are made and marketed in large variety.
Their composition and food value seem to have been studied
much less frequently chan many other commercial food products
notwithstanding the fact that a knowledge of their real value
would seem to be particularly important as they are chiefly
recommended by the makers for use in infant feeding and in
invalid dietetics.

The composition and true nutritive value of a number of pro-
prietary foods and food products are discussed in a paper by
Graham Lusk (Jour. Amer. Med. Assoc, 49 (1907), No. 3, pp.
201, 202, 270), dealing with the general subjecc of the nutritive
value of such foods.

D. L. Edsall (Jour. Amer. Med. Assoc, 54 (1910), No. 3, pp.
193-196) also discusses this general question, as has J. Rowland
(Jour. Amer. Med. Assoc, 54 (1910), No. 3, pp. 196-201), who
pays particular attention to predigested foods.

A published paper gives data regarding predigested foods and
similar goods (Jour. Amer. Med. Assoc, 48 (1907), pp. 1612-

162 Original Communications: Eighth International [vol.

1614, 1694; 49 (1908), pp. 1294, 1295; abs. in Chem. Abs., 2
(1908), No. 12, pp. 1740, 1741).

G. F. Richmond and W. E. Musgrave (Philippine Jour. Sci.,
3 (1909), No. 2, pp. 87-90) report a study of the composition of
malted milk, particularly its fat content, which was found to be
8.18 per cent.

Experiment station investigators have given much attention
to the breeding of cereal crops, the influence of fertilizers on com-
position, and other related questions. This work is perhaps
more appropriately considered in coimection with agricultural
chemistry than with nutrition, though some of it, notably that
with wheat and with corn, has an obvious relation to questions
of human nutrition.

It is interescing to note that comparatively wide variations
are observed in the composition of light and heavy kernels of
wheat of the same variety, in the grain from well developed and
imperfectly developed ears of corn, and in the kernels in different
parts of the ear.

In a study of the improvement of com, by A. M. Soule and
P. 0. Vanatter (Virginia Sta. Bui. 165, pp. 91-185, figs. 48), it
was observed that many of the best yielding ears did not have
as high a protein content as the undesirable ones.

C. L. Penny (Delaware Sta. Rpt. 1904-1906, pp. 13-33)
found a wide range in protein content, the minimum being 6.25
in one crop and the maximum 12.69. The smaller kernels at
the end of the ear were found to contain on an average 0.3 per
cent less protein than the large and well formed kernels.

These matters have been extensively studied at the Illinois
Experiment Station. In a report of investigations regarding
ten generations of com breeding, L. H. Smith (Illinois Sta. Bui.
128, pp. 457-575, figs. 2) summarizes data covering the range in
protein and fat content. The results obtained show that start-
ing with a single variety it was possible in ten generations to
increase the protein content "from 10.92 per cent to 14.26 per
cent, a gain of 3.34 per cent, while by breeding in the opposite
direction it has been possible to reduce the protein content from
10.92 to 8.64 per cent, a reduction of 2.28 per cent, making a
total difference between the two strains of 5.62 per cent. It is

xviii] Congress of Applied Chemistry 163

further shown that the high-oil com has increased from 4.70 per
cent to 7.30 per cent of oil, while a low-oil com has decreased
from 4.70 per cent to 2.66 per cent, the difference between the
two strains in 1906 being 4.71 per cent.

"High protein and low protein seed were planted together on
one plat and high-oil and low-oil seed on another. These plats
were continued for 3 years, and the results secured did not indi-
cate that the soil influences the protein or the oil content.

"A study of the secondary effects produced by selection to
change the composition of the grain indicated that the change
in the composition of the grain has produced no very marked
effect upon the composition of other parts of the corn plant."

The composition of corn and corn products, including green
com, their nutritive value and place in the diet, and similar ques-
tions have been discussed in a popular summary by C. D. Woods
(U. S. Depc. Agr., Farmers' Bui. 298, pp. 40, figs. 2), published
in conneccion with the nutrition investigations of the Ofl&ce of
Experiment Stations, which contains some individual work re-
garding the composition and digestibility of hulled com and
com bread and some work regarding the composition of pop-
corn popped and unpopped.

Alice R. Thompson (Hawaii Sta. Rpt. 1908, pp. 51-58), of
the Hawaii Experiment Station, has reported the results of
studies of Japanese rice and Hawaiian-grown rice, both polished
and unpolished, and rice paddy and straw from imported and
Hawaiian rice and from rice grown under different conditions,
the nitrogenous constituents being determined in every case,
and proximate and ash analyses in the case of rice grain and rice
straw and paddy.

Little variation was noted in the chemical composition of the
different varieties of rice, and the author is of the opinion that the
claim for superiority of Japanese imported over Hawaiian-grown
rice is not substantiated so far as nutritive value is concemed.

Comparisons of the analyses of polished and unpolished grain
showed that the unpolished rice contained about four times as
much fat as the polished, as well as more protein, crude fiber,
and ash. Practically all the nitrogen of the rice grain was found
to be proteid nitrogen.

164 Original Communications: Eighth International [vol.

The question of the wholesomeness of polished and unpolished
rice and the more specific question of the possible relation of
pohshed rice to beri-beri are matters which have been given much
experimental study in recent years as a part of the general ques-
tion of the possible connection between the presence or absence
of particular mineral constituents, protein radicals, or other con-
stituents and the occurrence of the disease.

The matter of the possible relation of rice to beri-beri is of
particular importance in the regions of the Orient where rice is
the principal carbohydrate foodstuff, so naturally the question
has been studied as a part of the scientific work undertaken by
the Philippine Department of Science. H. Aron and F. Hoc-
son (Biochem. Ztschr., 32 (1911), No. 3-4, pp. 189-203), in an
investigation on rice as a foodstuff, have reported experimental
studies in which the balance of income and outgo of nitrogen was
determined on a rice diet supplemented by other foods chiefly of
vegetable origin, including such material as bananas, rice polish,
and phytin.

Analyses of a large number of samples showed that relatively
more phosphorus than nitrogen was lost by polishing rice. The
unpolished rice contained on an average from 0.7 to 0.8 per cent
P^O^, undermilled rice from 0.4 to 0.6 per cent, and overmilled
rice from 0.15 to 0.4 per cent.

The authors conclude thao an exclusive rice diet will not sup-
ply protein enough to meet man's demands, and that therefore
it must be supplemented by vegetable, or better, animal foods
rich in protein. Such a mixed diet is satisfactory from an hy-
gienic standpoint, provided the rice has noc lost too much phos-
phoius by overmilling or polishing. From their experimental
studies they conclude further that, for a man weighing 50 kg.,
a diet made up of rice supplemented by vegetable foods must
contain at least 75 gm. protein in order to meet hygienic require-
ments, and that a diet of rice supplemented by fish or meat must
contain at least 65 gm., of which at least | is supplied by animal
foods.

Information gained from practical experience with beri-beri
and unpolished rice in the Philippines was summarized by V. G.
Heiser (Philippine Jour. Sci., B. Med. Sci., 6 (1911), No. 3, pp.

xviii] Congress of Applied Chemistry 165

229-233), particularly regarding the efforts which have been
made to encourage the local use of unpolished rice and the suc-
cess which has attended it.

For purposes of convenience, "a rice containing less than 0.4
per cent of phosphorus pentoxid is regarded as polished and that
which contains a greater percentage of phosphorus pentoxid as
unpolished rice."

An attempt to secure legislation regarding the use of unpol-
ished rice in the Philippines is briefly discussed.

The question of cotton seed as human food has been consid-
ered by G. S. Fraps (Texas Sta. Bui. 128, pp. 5-15), who reports
analyses of cotton-seed flour, cotton-seed flour bread, and other
cotton-seed bakery products. The general conclusion is that
cotton-seed flour is rich in protein and that it may be used alone
or mixed with wheat flour for the preparation of appetizing foods.
In the author's opinion, there is no reason to believe that cotton-
seed flour will not prove a wholesome product when used in small
amounts.

In his discussion, the author draws attention to the fact that
cotton seed has more or less proved harmful when used as food
for domestic animals, particularly pigs, but he is of the opinion
that the quantities likely to be used would not prove harmful to
man. Nevertheless, he cautions against using too large amounts.

It is interesting to note that F. Russell (Ann. Rpt. Bur. Amer.
Ethnol., 26 (1904-5), pp. 66-92, figs. 7) states that cotton seed
was formerly used as foodstuff by the Pima Indians of southern
Arizona.

The widespread interest at the present time in the possibility
of using cotton-seed meal as a food for man lends a special in-
terest to the investigations which have been undertaken to de-
termine the reason why it proves harmful to domestic animals,
particularly pigs, when fed a considerable time in fairly g'^nerous
quantities. Such studies will probably be referred to in detail
elsewhere. It may be noted here that it seems to be the case
that the renal disturbances or other pathological conditions ob-
served when it is fed to pigs may be postponed or even in some
cases materially lessened by feeding a large proportion of green
fodder with the cotton seed.

166 Original Communications: Eighth International [vol.

Interesting investigations on the general question of the pois-
onous properties which cotton seed sometimes exhibits when fed
to farm animals have been carried on in the Bureau of Animal
Industry and reported by A. C. Crawford" (U. S. Dept. Agr.,
E. S. R., 22 (1910), No. 6, pp. 501-505). His conclusion is that
the poisonous principle is not an alkaloid but probably an inor-
ganic compound, namely, a salt of pyrophosphoric acid. Phos-
phoric acid has long been known to be present in cotton-seed
meal in considerable quantity, and has been suggested as having
a possible relation to its toxicity, but the methods of study fol-
lowed have not been such as to bring out this relationship or
lend support to the hypothesis.

The conclusions advanced are supported by a large amount of
data from a systematic series of laboratory studies and physiolo-
gical tests and have been further confirmed by feeding experi-
ments with dogs carried on by the Bureau of Animal Industry
which were not reported in the preliminary account of the work.

Not all cotton seeds exhibit poisonous properties, particu-
larly being influenced in this respect by variety and by method
of cultivation.

To quote from Dr. Crawford's conclusions, "the chief poison-
ous principle in certain cotton-seed meals is a salt of pyrophos-
phoric acid. In some, this salt seems to be a simple one, pre-
sumably inorganic, while in others, it is more complex, perhaps
an organic one. Probably this difference in the combinations of
pyrophosphoric acid may aid in explaining the variation in tox-
icity of different meals. In certain cotton-seed meals one would
expect to find salts of metaphosphoric acid entering into this
action. To be harmful, the pyrophosphates must be in such
a form that they can be absorbed, or the phosphoric acid ionized
in the gastro-intestinal tract. The harmlessness of certain cot-
ton seeds and meal is mainly due to the fact that in them the
phosphoric acid exists largely, if not entirely, as a compound of
ortho, and not as one of the other phosphoric acids. Small
amounts of pyrophosphates can apparently be borne without
injury. The amount of the salt which may be permitted in cot-
ton-seed meal should be determined."

("Jour. Pharmacol, and Expt. Ther., 1 (1910), No.5, pp. 519-548).

xviii] Congress of Applied Chemistry 167

Many summaries of data regarding the composition of foods
have appeared, such work not infrequently forming a part of
treatises on food and nutrition.

A set of fifteen colored food charts, prepared by C. F. Lang-
worthy (U. S. Dept. Agr., Office Expt. Stas. Food and Diet Chart
15), has been issued in connection with the nutrition investiga-
tions of the Office of Experiment Stations, which are designed to
show graphically the composition and energy value of the common
food materials and to summarize some general data regarding
the functions and uses of food.

Special Studies op Ash, Protein, and Other Food Con-
stituents

No new products of particular importance have appeared dur-
ing the period under consideration in this summary, though
many of more or less general importance have been studied,
including dairy products, fruits, meats, cereal grains and other
materials. Methods of analysis as usual have received a great
deal of attention.

Much work has been reported in connection with inspection of
food under government and state pure food laws. No attempt
can be made here to summarize this. As taken in connection
with other pure food work, it constitutes a subject in itself.

In addition to studies of the composition of food already cited,
a number of investigations have been reported which have to do
with some detailed study of food constituents. For instance, the
solubility relations of milk sugar, the vapor pressures of saturated
solutions of hydrated milk sugar, the influence of concentration
on the equilibrium between the forms of milk sugar, and other
similar questions were studied by C. S. Hudson (Jour. Amer.
Chem. Soc, 30 (1908), No. 11, pp. 1767-1783, figs. 2).

A bulletin by E. B. Forbes (Ohio Sta. Bui. 207, pp. 23-52),
of the Ohio Station, on the balance between inorganic acids
and bases in animal nutrition, endeavors to show the bearing
on practical animal nutrition of the relationship between those
mineral elements of our foodstuffs and of living animal tissues,
which in the body give rise to inorganic acids, and the various

168 Original Communications: Eighth International [vol.

means at the disposal of the animal for accomplishing protection
from these acids through effecting their neutralization. The rela-
tion of ash constituents to human nutrition in general is also
considered. The investigations are reviewed in detail and a
number of general deductions are drawn.

The available alkali in the ash of human and cow's milk in its
relation to infant nutrition was studied by J. H. Kastle (Amer.
Jour. Physiol., 22 (1908), No. 2, pp. 28^308). The salient
points of difference between the ash of the two kinds of milk, the
author points out, are: "Human milk contains relatively more of
its mineral matter in utilizable form than cow's milk; it can supply
the organism of the child with relatively larger amounts of
available alkali in proportion to the proteid than cow's milk; it
contains much less proteid; and it contains a more readily absorb-
able variety of fat."

The nature of the chemical combinations of potassium in the
tissues was investigated by W. Koch and C. C. Todd (Abs. in
Jour. Biol. Chem., 9 (1911), No. 2, pp. XV, XVI; Proc. Amer.
Soc. Biol. Chem., 2 (1910), No. 1, pp. 9, 10). The results thus
far obtained indicate that "sodium and potassium phosphatid
compounds exist in all the tissues of the body and are probably
of much more importance than the hitherto assumed ion-protein
combination."

H. S. Grindley and E. L. Ross (Jour. Biol. Chem., 8 (1910),
No. 6, pp. 483— i93) have studied the determination of organic
and inorganic phosphorus in meats. Judging from the data
which they recorded, it appears that the coagulation of the pro-
tein of the aqueous extracts of flesh by heat does not change
organic phosphorus to the inorganic form to any appreciable
extent.

The subject has also been studied by P. F. Trowbridge and
Louise M. Stanley (Jour. Indus, and Engin. Chem., 2 (1910),
No. 5, pp. 212-215; abs. in Analyst, 35 (1910), No. 412, p. 311).
The proportion of soluble organic phosphorus in total soluble
phosphorus in meat was found to vary considerably in different
animals and in different parts of the carcass of the same animal.
The lowest recorded value (26 per cent) was observed with an
emaciated steer, and the highest (91 per cent) with a fat show

xviii] Congress of Applied Chemistry 169

steer. " During cooking, a progressive splitting up of the organic
phosphorus compounds takes place, and in well-cooked meats
practically the whole of the phosphorus is present in inorganic
combination."

Data are presented by C. K. Francis and P. F. Trowbridge
(Jour. Biol. Chem., 7 (1910), No. 6, pp. 481-501; 8 (1910), No.
1, pp. 81-93) regarding investigations of the kind and amount
of phosphorus present in beef cattle in different conditions of
fatness. The results were not uniform enough to warrant general
deductions. No relation was evident between phosphorus and
total ash.

The question was also studied with reference to the kind of
phosphorus present in different cuts.

"The round cut of beef contains more phosphorus, in forms
which are soluble in cold water than any of the other cuts.
Phosphorus is found chiefly in the muscular or connective tissue;
the fats contain but little. The flesh of a thin animal contains
more soluble phosphorus than that of a fat animal. The quan-
tity decreases with increasing fatness even when it is expressed
on a moisture and fat-free basis."

The nature of the phosphorus compounds of the brain, both
normal and diseased, was studied by W. Koch (Jour. Amer. Med.
Assoc, 52 (1909), No. 18, pp. 1381-1383), the work in considerable
part dealing with the phosphorus supply in the diet. The phos-
phorus required for the growth of the brain the author concludes
is amply supplied by the phosphorus of our daily diet. "If
desired, the addition of phosphorus-rich foods, such as eggs,
sweetbreads (pancreas), liver, and some meats, can be made to
meet further requirements, and will far exceed in amount the
phosphorus obtained in less natural form from the prescribed
doses of any of the various drugs in commercial use. The use of
such foods is, however, limited by their richness and their tend-
ency, on account of their rich fat content to interfere with gastric
digestion.

"As far as the nervous system is concerned, the addition to
the diet of commercial phosphorus compounds, such as hypo-
phosphites, glycerophosphate, phytin, lecithin, etc., is to be
discouraged because, in the first place, there is no conclusive

170 Original Communications: Eighth InternaUonal [vol.

evidence that they have any effect on the growth of the brain,
and, second, the amount usually recommended means only a
very insignificant addition to the amount of phosphorus (even
in its special forms such as lecithin) taken with the daily food/'

The relation of brain phosphatids to tissue metabolites was
studied by W. Koch and W. W. Williams (Jour. Pharmacol,
and Expt. Ther., 2 (1910), No. 3, pp. 253-264). Some of the
conclusions follow, which were drawn from experiments with
substances which may be regarded as of food value to the
tissues, including amino acids, glycocoU and glucose, and with
substances having a characteristic physiological action, includ-
ing among others adrenalin, caffein, and theobromin :

"The changes in state of aggregation of lecithin produced by
sodium chlorid are the result of the independent action of the
sodium and chlorin ions, whose effects are in opposite directions.
Below the concentration of a physiological salt solution (0.12
molecular) the action of the chlorin ion, which decreases the
state of aggregation of the lecithin, predominates. Above the
concentration of a physiological salt solution, the action of the
sodium ion, which tends to increase the state of aggregation of
lecithin, comes more and more into prominence.

"It has been suggested that, when the phenomenon of chlorid
retention occurs, some change has taken place in the state of
aggregation of the cell lipoids which allows this action of the
chlorin ion to predominate to a still greater extent.

"Ammonia and bile salts possess the power of altering the
physical state of aggregation of lecithin to such an extent as to
permit of the conclusion that they can be of functional sig-
nificance in altering the permeability of cell membranes. . . .
"The ability of the tissue metabolites to combine with lecithin,
as measured by the changes in the physical state of aggregation
produced by their presence, is in some cases considerable, in
other cases entirely lacking. Thus hypoxanthin, creatin, creat-
inin, adrenalin, and ammonia salts show evidence of combi-
nation. Inosit is doubtful and urea is negative.

"The amino acids show varying powers of combination.
The dicarboxy-acids, like acids in general, tend to increase the
state of aggregation of lecithin."

xviii] Congress of Applied Chemistry 171

In connection with the nutrition investigations of the Depart-
ment of Agriculture, H. C. Sherman (U. S. Dept. Agr., Office
Expt. Stas. Bui. 185, pp. 80) studied iron in food and its functions
in nutrition, and reported the results of three metabolism experi-
ments in which the balance of income and outgo of nitrogen and
iron and other mineral constituents was determined, as well as
the results of two dietary studies undertaken with special refer-
ence to the iron content of the food consumed. Estimates were
also made of the amounts of iron taken per man per day in 20
dietary studies made in connection with earlier nutrition investi-
gations of the Office of Experiment Stations.

"Increase of iron," it is pointed out, "in the diet without a
corresponding increase of protein is readily accomplished by the
use of vegetable, fruits, and the coarser mill products of the
cereal grains. In the experimental dietary here reported, the
free use of such foods with milk but without meat or eggs resulted
in an increase of 30 per cent in the iron content of the diet, while
the protein, the fuel value, and the cost remained practically the
same as in the ordinary mixed diet obtained under the same
market conditions."

In continuation of the work on mineral constituents, H. C.
Sherman, A. J. Mettler, and J. E. Sinclair (U. S. Dept. Agr.,
Office Expt. Stas. Bui. 227, pp. 70) have studied calcium, mag-
nesium, and phosphorus in food and nutrition, reporting the
results of 6 experiments on the metabolism of these constituents
and a study of the amounts present in typical American dietaries.
In general, the investigations show the importance in the diet of
calcium, magnesium, and phosphorus and the possibility of
securing them by the use in proper proportion of ordinary food
materials.

To quote, "it is entirely feasible to increase largely the calcium
and phosphorus intake by making a more liberal use of milk in
the dietary. The same may, of course, be said of the various
milk products in which the calcium and phosphorus compounds
are largely or wholly retained, such, for example, as cheese,
junket, kumiss, buttermilk, or cream. This is probably the
simplest and more effective means of improving the dietary as
regards calcium and phosphorus compounds, without decreasing

172 Original Communications: Eighth International [vol.

its aooeptability or materially increasing its cost and with distinct
advantages in other directions."

The balance of acid-forming and base-forming elements in
foods, was studied by H. C. Sherman and J. E. Sinclair (Jour.
Biol. Chem., 3 (1907), No. 4, pp. 307-309). Peas, milk, and
prunes are the foods studied containing an excess of base-forming
over acid-forming elements. With beef, oatmeal, and ' entire
wheat grain the reverse was the case. It is obvious, the authors
note, that "by the free use of meats and breadstuffs on the one
hand or of fruits, vegetables, and milk on the other, the net excess
of acid or base introduced into the body through the food may
be varied at will within wide limits."

The very important work of T. B. Osborne and his associates
on the cleavage products of protein has been continued, the
studies reported having to do with the hydrolysis of excelsin
(Amer. Jour. Physiol., 19 (1907), No. 1, pp. 53-60, pi. 1); hordein
(Amer. Jour. Physiol,. 19 (1907), No. 1, pp. 117-124); legumin
from the pea (Jour. Biol. Chem. 3 (1907), No. 3, pp. 219-225);
glycinin from the soy bean (Amer. Jour. Physiol., 19 (1907), No.
4, pp. 468-474); the crystalline globulin of the squash seed
(Cucurhita maxima) (Amer. Jour. Physiol., 19 (1907), No. 4, pp.
475-481) ; amandin from the almond (Amer. Jour. Physiol., 20
(1908), No. 4, pp. 470-476); the proteins of maize {Zea mays)
(Amer. Jour. Physiol., 20 (1908), No. 4, pp. 477^93); gliadin
from rye (Amer. Jour. Physiol., 20 (1908), No. 4, pp. 494-499);
vicilin from the pea (Jour. Biol. Chem., 5 (1908), No. 2-3, pp.
187-195); legumelin from the pea (Jour. Biol. Chem., 5 (1908),
No. 2-3, pp. 197-205) ; fish muscle (Amer. Jour. Physiol., 23 (1908)
No. 2, pp. 81-89) ; viteUin from the hen's egg (Amer. Jour. Physiol,
24 (1909), No. 1, pp. 153-160); muscle of scallop (Amer. Jour.
Physiol., 24 (1909), No. 1, pp. 161-169) ; crystalized albumen from
hen's egg (Amer. Jour. Physiol., 24 (1909), No. 2, pp. 252-262); ox
muscle (Amer. Jour. Physiol., 24 (1909) No. 5 pp. 437-446); casein
(Jour. Biol. Chem., 9 .(1911), No. 3-4, pp. 333-353); and
wheat gliadin (Jour. Biol. Chem., 9 (1911), No. 5, pp. 425-
438).

These very important invesagations are too extended for
summary here.

xviii] Congress of Applied Chemistry 173

Supplementing his work on the cleavage products of protein
T. B. Osborne and H. G. Wells (Jour. Infect. Diseases, 8 (1911),
No. 1, pp. 66-124) have studied the biological reactions of the
vegetable proteins, using the globulin from castor bean, flax seed,
and squash seed, edestin from the hemp seed, excelsin from the
Brazil nut, proteins from the cocoanut, legumin from the vetch,
legiunin and vicilin from the pea^ vignin from the cowpea,
glycinin from the soy bean, gliadin from wheat and rye flour,
hordein from barley, and zein from maize. All of these were
found to produce typical anaphylaxis in sensitized animals, the
condition possessing all of the characteristics which are present
when anaphylaxis is produced with serum or other animal sub-
stances containing soluble proteins.

It was found that considerable differences in toxicity were pro-
duced by the various proteins. "The most toxic proteins, as
measured by the frequency of severe and fatal reactions, were the
globulin of the squash seed, vignin, excelsin, and castor-bean
globulin, which usually caused death when given in 0.1 gm. doses
to properly sensitized animals. Edestin caused the least severe
reactions of any of the proteins, while hordein and glycinin seldom
caused fatal reactions; nevertheless, the minimum sensitizing and
intoxicating doses of edestin and squashseed globulin are essen-
tially the same."

The expeiiments showed, furthermore, that where continuous
feeding was done with the proteins, the guinea pigs became
immune to the proteins and could not be sensitized to them.
There was a marked specificity shown within certain limits by the
proteins, and a close similarity, if not identity, of the legumins of
the pea and vetch and the close relation to the vicilin of the pea
was shown by the interaction of these proteins. The probable
identity of the gliadin from wheat and rye, or at least their near
reaction, was also established. "In some instances doubtful
results were obtained, for example, with some guinea pigs castor-
bean globulin and flax-seed globulin interacted strongly, while
with others similarly treated, no reactions were obtained."

The structure of proteids, enzyms and their relation to biological
problems, and related questions are discussed in a paper by R. H.
Chittenden (Science, n. ser., 27 (1908), No. 685, pp. 241-254).

174 Original Communications: Eighth International [vol.

D. D. Van Slyke and P. A. Levene (Proc. Soc. Expt. Biol, and
Med., 6 (1908), No. 1, pp. 11-13) have reported studies of the
cleavage products of plastein, the "protein-like substance or
substances precipitated from concentrated albumose solutions
by the action of enzyms." Their results "indicate that the
plastein is related to the higher albumoses, and apparently,
from the resistance of alkali, to the antialbumoses rather than
to the native proteins."

T. B. Robertson (Jour. Phys. Chem., 13 (1909), No. 6, pp.
469-489) reported data which showed that the concentration of
casein solutions can be very accurately studied by determining
their refractive indices. His investigations are discussed at
length. He later reported the results of studies of the refractive
indices of solutions of certain proteins including the para-nucleins
(Jour. Biol. Chem., 8 (1910), No. 4, pp. 287-295), serum globulin
(Jour. Biol. Chem., 8 (1910), No. 6. pp. 441-448), casen in alcohol-
water mixtures (Jour. Biol. Chem., 8 (1910), No. 6 pp. 507-511),
and gliadin (Jour. Biol. Chem., 9 (1911), No. 3-4, pp. 181-184).

Cooking in its Relation to Nutritive Value.

The chemical changes involved in cooking processes have been
investigated with a considerable number of materials.

Some data regarding army rations, field ranges, ovens, fireless
cookers, the kitchen touring car, portable gas cooker for army
use, etc., are included in a report of the U. S. Commissary General,
H. G. Sharpe (Rpt. Commis. Gen. [U. S. Army], 1909, pp.
11-15).

Suggestions for a diet kitchen equipment, particularly with refer-
ence to naval hospitals, are presented in a paper by W. Wierzbicki
(U. S. Naval Med. Bui., 4 (1910), No. 2, pp. 161-163, dgms. 2).

Studies of housekeeping efficiency as a private enterprise, by
C. Barnard (Housekeeping Expt. Sta. [Conn.] Bui. 11, pp. 20,
pis. 3) form the basis of a discussion of the increased efficiency
through correct house planning, the use of conveniences and labor
saving devices, the elimination of needless work, and similar
questions. Some data are recorded regarding the labor involved
in performing a definite household task by different methods.

xviii] Congress of Applied Chemistry 175

Studies of the supposed connection between protein coagula-
tion and the heat shortening of muscles were reported by E. B.,
Meigs (Amer. Jour. Physiol., 24 (1909), No. 1, pp. 178-186
dgms. 6), and are interesting not only from the standpoint of
physiological chemistry but also because of their possible bearing
on the changes which take place in animal foods during cooking
processes. The facts reported, as the author points out, do not
preclude "the possibility that the precipitation of protein from
its solutions and the shrinkage of animal tissues under the influ-
ence of heat may be fundamentally more or less similar processes.
They do show, however, that the shortening of striated muscle
at temperatures above 50° is independent of the coagulation
of myogen, and they make it seem probable that the heat short-
ening of most animal tissues is dependent, not on the aggregation
of the particles of coagulable protein, but on some other process."

Elizabeth C. Sprague and H. S. Grindley (Univ. 111., Univ.
Studies, 2 (1907) No. 4, pp. 37, pis. 4, dgms. 10) studied the
cooking of beef with a view to formulating methods which would
give imiform results. In connection with this work, tempera-
tures were recorded of the interior of the beef during cookery.
If the temperature ranges from 55 to 65°C., the beef will be imder-
done or rare and red in color. At a temperature of 65 to 70°,
it sill be medium underdone, and at a temperature of 70 to 80°,
it will be well done.

In connection with his work showing the palatability and
wholesomeness of spleens, and studies of their preparation for the
table, E. T. Williams (Amer. Med., n. ser., 2 (1907), No. 9, pp.
522, 523) points out that although the raw spleens do not
keep well, the cooked material, particularly boiled, has excellent
keeping qualities. Attention is directed to the high iron and
phosphoric acid content of spleens.

The economical use of meat in the home and many questions
which have to do with the nutritive value of meat and the pre-
paration of meat for the table, have been discussed in a popular
summary by C. F. Langworthy and Caroline^L. Hunt (U. S.
Dept. Agr., Farmer's Bui. 391, pp. 43 II).

The question of cooking the cheaper cuts of meat is considered
on the basis of the author's experimental study of the problem,

176 Original Communications: Eighth International [vol.

by C. Barnard (Housekeeping Expt. Sta. [Conn.] Bui. 6, pp. 17,
dgm. 1).

The results of an extended series of artificial digestion experi-
ments on starch of different sorts as affected by cooking were
reported by Edna D. Day (U. S. Dept. Agr., Office Expt. Stas.
Bui. 202, pp. 42, figs. 6) in a bulletin of the Office of Experiment
Stations. Different sorts of ferments were used with potato,
wheat, com, and other starches. The conclusion was reached
that potato, arrowroot, and probably tapioca and sago starches,
are made not more easily digestible by long continued cooking,
while the reverse is true wioh cereal starches, though the changes
occur very slowly. In general, the experimental data reported
are discussed with reference to household problems.

The effect of cooking on cellulose was studied by Edna D. Day
(Jour. Home Econ., 1 (1909), No. 2, p. 177), who did not find
that cell walls of potatoes when boiled or baked are ruptured,
as is generally seated to be the case. When cells from cooked
potatoes were examined, it was found that the middle lamella
which holds the cells together had dissolved and that the cells
had separated from each other, but the cell walls were not
ruptured. "If, however, saliva is added to these unbroken
cells, the starch filling them is very quickly digested, as shown
by the fact that they no longer give the blue color with iodin,
proving that the breaking of the cell wall is not at all essential
for ease of digestion."

Studies of the etiology of pellagra reported by W. H. Buhlig
(111. Bd. Health Mo. Bui., 5 (1909), No. 7, pp. 417-435, figs. 2),
particularly with reference to the possible connection of Indian
com with this disease, did not lead to definite results. In con-
nection with culture tests with moldy corn, some cooking tests
were made, as certain molds are known to be resistant to heat,
and the idea has been advanced thaD such enzyms may survive
cooking. Com meal mush and hominy made in the usual way,
by boiling about 2 hours, was found to be sterile.

In other experiments carried on in the same public institution
as Buhlig's work, the question was further studied by J. F. Siler
and H. J. Nichols (111. Bd. Health Mo. Bui., 5 (1909), No. 7,
pp. 437-478, figs. 8), who repeatedly found in corn meal and

xviii] Congress of Applied Chemistry 177

hominy a spore-bearing bacterium which survived steaming for
2 hours.

A brief note on the effects of adding sugar to acid fruit at the
beginning and end of the cooking period was published by Edna
D. Day (Jour. Home Econ., 2 (1910), No. 1, p. 94), as the result
of tests with cranberries, grapes, and apples, and also of the com-
parative sweetness of solutions of the same strength of cane sugar
and a mixture of levulose and dextrose.

The conclusion is reached chat "in cooking such fruits as
apples, cranberries, and grapes, while the product is slightly
less sweet if the sugar is added at the beginning than it is if it
is added at the end, still the difference is too small to be of
practical importance."

The problems of cookery at high altitudes (diminished air
pressure) are discussed on the basis of experiments, by Mrs. A.
Anderson (Boston Cooking-School Mag., 14 (1910), No. 8, pp.
372, 373, XVI, XVIII, XX). Data on this subject have been
summarized in a recent paper (Jour. Home Econ., 3 (1911),
No, 2, pp. 176-178).

According to experiments briefly reported by Olive G. Patter-
son and Clara C. Benson (Jour. Home Econ., 2 (1910), No. 6,
pp. 656, 657) on the setting of gelatin, this material may be freed
from its mineral matter and tyrosin-holding impurities without
affecting the gelatinizing power of its solutions. Boiling for 1
hour did not prevent gelatinization though long-continued boiling
diminished it. With respect to the effect of citric and acetic
acids, it was found that 4 per cent gelatin solutions containing
citric acid to a concentration of 1 per cent would gelatinize in the
cold after 15 minutes' boiling, but that after 10 minutes' boiling
of a 3 per cent solution wich 0.5 per cent citric acid the gelatin-
izing power had considerably decreased.

The question of bread has received less attention from inves-
tigacors than it did a few years ago.

The relation of yeast to flavor in bread has been studied
experimentally by Ruth A. Wardall (Jour. Home Econ., 2
(1910), No. 1, pp. 75-91), who concludes that the flavor of bread
can not be determined by yeast and possibly is not even affected
by it.

178 Original Communications: Eighth International [vol.

Since for obvious reasons the time allotted for fermenting
bread is short, she regards it as quite possible that an insufficient
opportunity to develop flavor is given.

Some experiments were also made upon the effects of adding
malt extract to bread dough.

The leavening agent in salt-rising bread was studied experi-
mentally by Winona Woodward (Jour. Home Econ., 3 (1911),
No. 1, pp. 100, 101), who isolated an organism which was not a
yeast.

H. A. Kohman (Nat. Assoc. Master Bakers [Proc], 13 (1910),
pp. 29-37, fig. 1) studied salt-rising bread making, reaching che
conclusion thao the fermentation is due to a bacterium and not
a yeast. The bacterium was isolated and studied in pure culture.

Among general discussions of bread may be mentioned a paper
by M. E. Jaffa (Nat. Baker, 14 (1909), No. 166, pp. 52, 54),
which devotes considerable attention to the use of raisins in
bread making.

A large amount of data regarding the character and nutritive
value of bread of different sorts and similar topics is included in a
popular summary, entitled "Bread and Bread Making," by
Helen W. Atwater (U. S. Dept. Agr., Farmer's Bui. 389, pp. 47,
figs. 7).

A large amount of data has been reported regarding the prin-
ciples and practice of ice cream making, by R. M. Washburn
Vermont Sta. Bui. 155, pp. 92, dgm. 1). The work is based on an
exhaustive study of the subject, particularly from a commercial
standpoint.

The question of cooking naturally involves the relative value
of different kinds of equipment and other similar topics.

Labor and money-saving appliances are discussed in a publi-
cation of the American School of Home Economics (Bui. Amer.
School Home Econ., Ser. 1, 1908, No. 11, pp. 47, figs. 54), and
numerous other discussions have appeared in journals and
reports which have to do with such topics.

The matter of fireless cookers has been studied by a number of
investigators.

Fireless cookers'' of special construction have been tested in
connection with experiments on the preparation of food made

xviii] Congress of Applied Chemistry 179

by the subsistence department of the U. S. Army. This work
and data regarding foods supplied in the Philippines are
reported by H. G. Sharpe (Rpt. Commis. Gen. [U. S. Army],
1907, pp. 10-14).

The value of different materials for the construction of fireless
cookers and the effects of amounts and density of material upon
the conservation of heat were studied experimentally by Ellen
A. Huntington (Bui. Univ. Wis., No. 217, pp. 38, figs. 10), the
article as a whole being an interesting contribution to the subject.

The construction and use of the fireless cooker from a practical
standpoint have been studied by Caroline B. Lovewell, Frances
D. Whittemore, and Hannah W. I^yon (Topeka, 1908, pp. 211,
figs. 11).

A similar summary is also included in the volume dealing with
fireless cookers prepared by Margaret J. Mitchell (New York,
1909, pp. XII + 315, figs. 18).

Canning, Preserving, Handling and Storage

The question of canning and preserving food, the changes
brought about by cold storage, and related matters have been
studied by a number of investigators.

An exhaustive summary of data regarding the question of
storage of food products in the District of Columbia is contained
in che U. S. House of Representatives report (Report of hearings
on H. R. 16925, to regulate the storage of food products in the
District of Columbia — Washington: U. S. House of Representa-
tives Committee on District of Columbia, 1910, pts. 1-14, pp.
1-279).

The gases contained in swollen canned goods were studied
experimentally by F. 0. Tonney and J. B. Gooken (Amer. Food
Jour., 3 (1908), No. 6, pp. 20-23, figs. 3). In general, the
authors note that the presence of nitrogen indicates putrefaction
and carbon dioxid, fermentation, the two processes being often
found to be distinct from each other.

0 .W. Shaw (California Sta. Circ. 33, pp. 4-8, figs. 3) compared
cane sugar and beet sugar with a view bO determining whether
or not there is ground for the belief that beet sugar is inferior to

180 Original Communications: Eighth International [vol.

cane sugar for jelly making and preserving purposes. His con-
clusion was that such is not the case and that beet sugar is entirely
satisfactory for such purposes.

Information regarding packing house methods, shipping, keep-
ing quality, and similar subjects is included in a summary
of the results of field investigations in pomology by G. H.
Powell (U. S. Dept. Agr.,Bur. Plant Indus. [Circ], June 7,
1907, pp. 4).

In a volume by G. T. Hamel, entitled "Modem Practice of
Caiming Meats" (St. Louis, 1911, pp. 100, figs. 19, dgm. 1), the
theories of canning are discussed, equipment described, and recipes
and formulas given. The subject is considered from the stand-
point of the small plant as well as from that of the large estab-
lishment.

The principles of canning are discussed a;nd directions for
canning a large number of fruits and vegetables and for
pickling and preserving meats and fish are included in a bulletin
by G. McCarthy (N. C. Dept. Agr., Biol. Div., 1907, pp. 37),
designed for the use of housekeepers.

An extended summary of statistics regarding canning and pre-
serving fruits and vegetables, fish, and oysters was presented by
E. K. Ellsworth (Bur. of the Census [U. S.J Bui. 61, pp. 9-48),
in a pubUcation of the U. S. Census Bureau.

The culinary qualities of dehydrated eggs, fruits, vegetables,
and milk were reported upon by H. A. Dent (Navy Dept., Bur.
Supplies and Accts., Mem. Inform. Off. Pay Corps) [etc.J, No.
85, pp. 626, 627), of the U. S. Navy Department.

The general question of canning vegetables in the home is
discussed in a popular summary prepared by J. F. Breazeale
(U. S. Dept. Agr., Farmers' Bui. 359, pp. 16, figs. 9), of the
Bureau of Chemistry.

A. W. Bitting (U. S. Dept. Agr., Bur. Chem. Bui. 119, pp. 37,
pis. 2, figs. 5) discusses the problem of catsup making in connection
with an experimental study of the spoilage of tomato catsup.

The cause of cloudy Hquor on peas was investigated by E. W.
Duckwell (Canner and Dried Fruit Packer, 29 (1909), No. 1,
pp. 34, 36), who reached the conclusion that it was caused by
starch from the peas and overheating in canning.

xviii] Congress of Applied Chemistry 181

A popular summary of data on canning peaches has been
published by H. P. Gould and W. F. Fletcher (U. S. Dept.
Agr., Farmers' Bui. 426, pp. 26, figs. 14), of the Bureau of
Chemistry.

A study of the preparation of sugared and dried pineapple has
been reported by H. C. Gore (U. S. Dept. Agr., Bur. Chem. Giro.
57, pp. 8, fig. 1).

An experimental study of packing prunes in cans, with satis-
factory results, was also reported by G. W. Shaw (California
Sta. Circ. 33, pp. 1-3).

Studies of jelly and jelly making have been reported by Nellie
E. Goldthwaite (Jour. Indus, and Engin. Chem., 2 (1910), No.
11, pp. 457-462, fig. 1). A number of small fruits were used as
well as orange juice, skins, and whole fruit.

According to the author's summary, "in what is usually a waste
product (the white inner skins of oranges and lemons) we have an
abundant source of pectin from which excellent jelly can be made
if properly acidified.

"It was noteworthy that the purest pectin yet prepared in
this research was obtained from oranges and lemons. It was
isolated . . . and was reprecipitated three times. By
long manipulation of the precipitated pectin (supported on a
very fine cloth suspended from the corners) the liquid was so
completely worked out of the substance that a powdery white
body, somewhat starch-like in appearance, was obtained. This
was dried in a current of dry hydrogen over sulphuric acid.

"Ash determinations of orange pectin so obtained showed less
than 0.5 per cent of ash — of lemon pectin about 3.5 per cent. .
. . No melting point of this pectin could be obtained, but the
substance, when out of contact with air, chars strongly at 170 C.
It is hoped to continue this work on the isolation and examination
of pure pectin."

A chemical-physical study of jelly maldng was carried on at the
Florida Experiment Station, by J. Belling (Florida Sta. Rpt.
1908, pp. CV-CIX), the investigations having particularly to do
with the influence of preliminary heating, final temperature, and
the percentage of water, sugar, and acid upon appearance and
quality of guava jelly.

182 Original Communications: Eighth International [vol.

"In boiling of guava jelly, some acid (the natural acid of the
ripe fruit) is absolutely necessary to change much of the sucrose
into invert sugar, and if this does not take place then the sucrose
crystallizes out. Too much acid (and probably too prolonged
boiling) seems to make the jelly sticky from the excess of invert
sugar, and also to alter the pectin so that it will not
gelatinize.

" The depth of color seems to be increased by additional amount
of acid, prolonged boiling, and higher temperature at which the
boiling is stopped."

f- Experiments of great value with reference to the handling and
marketing of fruits have been carried on at the Department of
Agriculture and elsewhere.

Many questions which have to do with the preparation for
shipment and marketing in fresh condition of fruits of different
sorts have been studied in a paper by A. V. Stubenrauch (U. S.
Dept. Agr. Yearbook 1909, pp. 365-374, pis. 3), of the Bureau of
Plant Industry.

The method of pre-cooling fruit for shipment is discussed by
G. D. Kellogg (Cal. Fruit Grower, 40 (1909), No. 1120, p. 1) in
comparison with results obtained by ordinary methods of shipment.

The problem of time and temperature in cold storage, partic-
ularly with reference to the Kieffer pear, was studied by G. H,
McKay (Proc. N. J. Hort. Soc, 32 (1907), pp. 127-135).

Experiments on the processing of persimmons to render them
nonastringent, carried on by H. C. Gore (U. S. Dept. Agr., Bur.
Chem. Bui. 141, pp. 31, pis. 3, figs. 5), though regarding as pre-
liminary, tend to show that carbonic acid gas may be substituted
for sake fumes for this purpose with Japanese persimmons, and
that, combined with the use of dry starch to prevent cracking of
the fruit during the processing, should lead to the perfection and
use of this method for the production of nonastringent persim-
mons which may be pared and eaten like an apple.

A. E. Vinson (Plant World, 10 (1«07), No. 11, pp. 259-262)
has studied exhaustively the composition of dates with special
reference to stages of ripening, the possibility of stimulating ripen-
ing, and related questions. As a part of his woik, a special study
of the endo- and ektoininvertase of the date isreported by A. E.

xviii] Congress of Applied Chemistry 183

Vinson (Jour. Amer. Chem. Soo., 30 (1908), No. 6, pp. 1005-
1020). The invertase of the date, he notes, remains "insoluble
in all ordhaary solvents throughout its green stages, but becomes
readily soluble on ripening. The change in the behavior of the
invertase towards solvents coincides very closely in point of time
with the passage of the tannin into the insoluble form."

The influence of chemicals in stimulating the ripening of fruits
has been studied further by Vinson (Science, n. ser., 30 (1909),
No. 774, pp. 604, 605), who found that dates could be success-
fully ripened by exposing them to a vapor of acetic acid for 12
or 15 hours. "At the end of this time they have become trans-
parent nearly to the seed and will then ripen naturally without
further treatment. The process can be accelerated by exposing
them to sunshine, or more rapidly by heating for some hours to
45°C. The process, it is anticipated, will permit the shipping of
dates green and ripening them at their destination as bananas
are now handled.

"After moderate treatment with acetic acid, the tannin of the
date has not yet become entirely insoluble but all astringency
disappears in the next few hours. The intracellular invertase,
however, passes into solution to quite an appreciable extent im-
mediately after the treatment, and probably other intracellular
or insoluble catalytic agents are released simultaneously."

The fresh ripe date is very soft and will not bear shipment.
The author believes that by treatment with acetic acid vapor,
it may be successfully ripened after shipment, a deduction of
much commercial importance.

Continuing his study of the stimulation of premature ripening
by chemical means, the author has studied the effect of many
other substances beside acetic acid vapor on the ripening of dates
and finds that a comparatively large proportion, including
among other proprionic acids, ethyl chlorid, chloroform, gasoline,
ether, acetone, and volatile oils. A. E. Vinson (Jour. Amer.
Chem. Soc, 32 (1910), No. 2, pp. 208-212).

Studies of the artificial ripening of dates by the aid of chemical
substances were later reported in greater detail by A. E. Vinson
(Arizona Sta. Bui. 66, pp. 403^35), and of ripening by incuba-
tion by G. F. Freeman (Arizona Sta. Bui. 66, pp. 437-456).

184 Original Communications: Eighth International [vol.

In a study of methods of canning meat with reference to pro-
per disposal of defective cans, C. N. McBryde (U. S. Dept. Agr.,
Bur. Anim. Indus. Rpt. 1907, pp. 279-296, fig. 1) draws a number
of general conclusions regarding putrefactive and fermentative
changes in the contents of cans, and similar matters.

Packing oysters with and without ice and similar questions
were studied by the Indiana State Board of Health, (Mo. Bui. Ind.
Bd. Health, 10 (1908), No. 11, pp. 134-136, fig. 1) with reference
to the pecuniary loss to the consumer and the melting of ice in
the oysters, analytical data being reported.

The possibility of preserving eggs with a number of substances,
including sodium silicate of different grades, was studied by R.
Berger (Jour. Indus, and Engin. Chem., 3 (1911), No. 7, pp.
493-495; Reprint, pp. 4; Pure Products, 7 (1911), No. 8, pp.
423-425), who reports data regarding the permeability of the
shells of preserved eggs and their loss in weight when kept in
the open air after preservation ia comparison with unpreserved
eggs.

Bacteriological studies which have to do with the infection
and preservation of eggs were carried on by G. H. Lamson,
Jr. (Connecticut Storrs Sta. Bui. 55, pp. 203-214, figs. 7) at
the Connecticut Storrs Station, with reference to the cause of
decomposition and sources of infection of eggs, the part played
by temperature, and the precautions to be observed in preserving
eggs.

The diastatic enzym of ripening meat was studied by A. W.
Peters and H. A. Mattill (Jour. Biol. Chem., 6 (1909), No. 2,
pp. XXIX, XXX). When muscle is autolyzed, the sugar
becomes greater, they conclude, provided the meat is fresh and
edible; otherwise, the amount diminishes.

The nature of so-called "black spots" on chilled beef was
studied by E. Klein (Meat Trades' Jour., 30 (1909), Nos. 1113,
p. 234; 1114, pp. 260, 261, figs. 2), who attributes much to the
growth of a fungus {Oidium carnis), which is described.

R. Hoagland (U. S. Dept. Agr., Bur. Anim. Indus. Rpt.
1908, pp. 301-314), of the Bureau of Animal Industry, has studied
the action of saltpeter upon the color of salted meat. The red
color of uncooked salted meat, to which saltpeter has been added

xviii] Congress of Applied Chemistry 185

as a preservative agent, is due, he concludes, to the presence of
NO hemoglobin which is formed by the action on hemoglobin of
nitric acid due to the reduction of nitrites within the meat.
The reduction of saltpeter to nitrites takes place in the meat
equally well in either an acid, or an alkaline medium. Neither
saltpeter nor nitrites exercise a color-preserving action in meat.
The brown color observed when meat is cured with an excessive
amount of saltpeter is due to the action of nitrites upon hemo-
globin.

In connection with a study of deterioration and commercial
preservacion of flesh foods, W. D. Richardson and E. Scherubel
(Jour. Amer. Chem. Soc, 30 (1908), No. 10, pp. 1515-1564)
report the results of a study of experiments on frozen beef, from
which the conclusion is drawn that no decomposition is shown
in frozen beef stored, for a long period, judging by the values
obtained for ammonia nitrogen, and hence that no bacterial
decomposition occurred in the stored meat. They conclude
further that the stored meat did not differ in flavor from the
fresh meat and that cold storage below -9°r. is an adequate
and satisfactory method for the preservation of beef for long
periods.

W. D. Richardson and E. F. Scherubel (Jour. Indus, and Engin.
Chem., 1 (1909), No. 2, pp. 95-102) also reported an extended
series of analyses of meat kept at room temperature with and
without preservatives and of meat stored at 2 to 4°C., and in
some cases afterward frozen and held at -9 to -12°C.

"While there are some contradictory figures in the analyses of
the samples which were frozen afucr being stored at 2 to 4°C.,
from the results the conclusion may fairly be drawn that freezing
of meats at -9 to -12°C. arrests bacterial decomposition, but can
not in any degree restore the product to its original condition."

The results of an extended study of the effects of cold storage
upon beef and poultry are reported by A. D. Emmect and H. S.
Grindley (Jour. Indus, and Engin. Chem., 1 (1909), Nos. 7, pp.
413-436; 8, pp. 580-597), as part of an experimental study of the
chemistry of flesh. The samples included uncooked refrigerated
beef stored for 22 days and for 43 days and frozen drawn and
undrawn fowl. Tests were also made of the relative losses in the

186 Original Communications: Eighth International [vol.

cooking of refrigerated beef held in cold storage for varying lengths
of time, and the chemical changes resulting therefrom. The
cooked meats from the sample stored 43 days were higher in their
moisture conteat than the sample stored 6 days and were therefore
juicier, higher in soluble and insoluble dry substance, in nitro-
genous, nonnitrogenous and total organic extractives, in fat, in
total ash, and in soluble inorganic, total soluble, and total phos-
phorus. "Further, the percentages of total nitrogen, insoluble
and total protein were practically the same as were those for the
samples from the 6-day storage meat. Therefore, the cooked
meats from the 43-day samples, judging from the chemical com-
position, were at least as nutritous as were those from the samples
stored for the shorter period of time."

The dietetic value of refrigerated foods as a whole is discussed
by S. Rideal (Cold Storage and Ice Trade Jour., 36 (1908), No.
4, pp. 32, 33) on the basis of experimental data. The action of
diastase, he concludes, is not entirely prevented by cold, but is
very much retarded. The tenderness and maturing of refiig-
eraoed meat he attributes not only to the action of sarcolactic
acid but also to the gradual and limited work of natural enzyms
(pepsin and trypsin) which cause a certain amount of pre-
digescion.

The changes which take place in chickens in cold storage have
been exhaustively studied by Mary E. Pennington (U. S. Dept.
Agr. Yearbook 1907, pp. 197-206, pis. 7), of the Bureau of Chem-
istry, which led to the general conclusion that both microscopic
study and the taste of the cooked fowl confirmed the fact that
microscopically visible degeneration does take place during long-
continued storage.

The effects of different methods of handling and storing poultry,
particularly with reference to the subject of drawing poultry,
and the storage of poultry and eggs have been discussed by Mary
E. Pennington (Ice and Refrig., 40 (1911), No. 2 pp. 59-62,
charts 6; Nat. Provisioner, 42 (1910), Nos. 4, pp. 16, 23, 24; 5,
pp. 23, 24; U. S. Dept. Agr., Bur. Chem. Circ. 64, pp. 42,
figs. 9).

A chemical study_of drawn and undrawn poultry kept in cold
storage was also reported by W. F. Boos (Aim. Rpt. Bd. Health

xvm] Congress of Applied Chemistry 187

Mass., 39 (1907), pp. 263-283), and a bacteriological examination
of such poultry by H. R. Brown (Ann. Rpt. Bd. Health Mass.,
39 (1907), pp. 285-336). According to Brown's conclusions,
"decomposition depends largely upon the presence of moisture
in the tissues, for moisture is absolutely essential to bacterial
growth. In freshly killed birds, ordinarily or properly drawn,
the surfaces quickly become dry. In cold storage birds, no
matter how they are drawn, the tissues will be moist, because
of the melting of the crystals of ice. If properly drawn, there
would be but few bacteria present capable of causing decom-
position."

Much attention has been devoted to the question of the relative
wholesomeness of drawn versus undrawn poultry, by E. W.
Burke (Amer. Food Jour., 3 (1908), No. 9, pp. 7-10).

A study of the effects of cold storage on eggs, quail, and chicken
was reported by H. W. Wiley, et al. (U. S. Dept. Agr., Bur. Chem.
Bui. 115, pp. 117, pis. 13), the general results being unfavorable
to this process when long continued.

The effect of low temperatures on ground chicken meat was
studied by H. W. Houghton (Jour. Indus, and Engin. Chem., 3
(1911), No. 7, pp. 497-505), in comparison with the original
chicken meat. The chemical changes which apparently take
place "are (1) slight variations in the case of moisture and other
ejrtract; (2) a small increase of ammonia, especially in the case of
the light chicken meat; (3) a decided increase of water-soluble
nitrogen, total solids, and organic extractives in the light chicken
meat, with a slight decrease of the same constituents in the dark
meat; (4) a decrease of coaguable nitrogen in both varieties of
chicken meat during the first 30 days, followed by a rise which
did not reach that of the fresh sample; (5) an increase of amino
acids in both kinds of chicken meat, with an increase and decrease
of the proteoses and peptones respectively in the light and dark
chicken meat."

Chicken fat has been studied extensively at the Bureau of
Chemistry. Mary E. Pennington and J. S. Hepburn (U. S.
Dept. Agr., Bur. Chem. Circ. 75, pp. 11) report the occurrence of
lipase in the crude fat of chickens, and find that it can resist
freezing for as long a period as 89 months, (pp. 1-7).

188 Original Communications: Eighth International [vol.

The oxidation of chicken fat by means of hydrogen peroxid was
studied by J. S. Hepburn (pp. 8-11), particularly with reference
to the effect of prolonged freezing.

"The changes in the fat of chickens during prolonged freezing
are similar to the changes called forth by oxidation of the fat with
hydrogen peroxid. The Hehner number and the saponification
number increase simultaneously, and aldehydes are formed.
The increase in saponification number may, therefore, be ascribed
to the formation of slightly lower homologues of the fatty acids
of fresh chicken fat, while the increase in Hehner number is
doubtless due to the formation of aldyhdes and ketones of high
carbon content. These changes in ^he chicken fat in situ are
probably produced by the action of enzyms."

The preparation of cod and other salt fish for the market was
studied by A. W. Bitting (U. S. Dept. Agr., Bur. Chem. Bui. 133,
pp. 63, pis. 6, figs. 4), of the Bureau of Chemistry, who also
reports the results of a study of the cause of reddening in fish.
This change was found to be due to a micro-organism, a remedy
being extreme cleanUness.

The bulletin gives information regarding the composition of
salt used in curing, losses in weight during curing, the amount of
salt taken up by fish marketed in different forms, and variations
in moisture and salt content due to season, style of packing, and
other conditions.

L. W. Thomas (North Dakota Sta. Spec. Bui. 24, pp. 179-194,
fig. 1) has reported a study of wrapped and unwrapped loaves,
with reference particularly to moisture content and keeping
quality. The general conclusions drawn are as a whole plainly
in favor of wrapping bread, though, as the author points out,
wrapping did not prevent loaves from becoming stale after 36
or 48 hours.

H. L. White continued this work by studying the moisture
and acidity of samples of wrapped and unwrapped bread. Ac-
cording to his summary, bread made under cleanly conditions
from good quality materials did not grow acid, when wrapped
or unwrapped, even after 108 hours. Bread wrapped while warm
and while hot showed a slight increase in acidity in the inside
of the loaf.

xviii] Congress of Applied Chemistry 189

C. A. A. Utt (Bui. Kans. Bd. Health, 7 (1911), No. 3, pp. 52-
60) has also reported tests of the effecc of wrapping bread upon its
quality. In general, the unwrapped bread, when kept for 4 or 5
days, lost about twice as much moisture as the wrapped loaf, while
the acidity remained practically the same. The wrapped bread
was in edible condition for twice the ordinary period.

As pointed out by H. G. Bell (Amer. Miller, 37 (1909), No. 4,
pp. 280, 281, fig. 1), ui a paper on stored flour, fungi and bacteria
are the chief destructive agencies. Flour of different grades was
studied with reference to the presence of bacteria and as pro-
tection against the growth of these low forms of Ufe, the author
suggests storage in well-lighted rooms.

The changes in the weight of stored flour and butter were
studied in detail by J. T. Willard (Bui. Kans. Bd. Health, 7 (1911)
No. 1, pp. 9-14). The greatest loss in flours stored for a year was
0.79 lb. per sack of about 48 lbs. Loss of weight in butter was
determined by the method of packing. Prints wrapped in parch-
ment paper and placed in paraffin cartons, packed in cases,
remained practically constant in weight. The loss in weight is
chiefly due to loss of moisture by evaporation or in other ways.

Changes which take place in the composition of unground
cereals during storage were studied by S. Leavitt and J. A. Le
Clerc (Jour. Indus, and Engin. Chem., 1 (1909), No. 5, pp. 209-
302). The investigations extended over 2 years. The results
demonstrated that "there is more or less change in all cereals
under the influence of aging. These changes seem to take place
whether the cereal is stored in the whole grain or is ground to a
fine powder before storage. In the latter case, however, the
changes take place more rapidly. We notice that the principal
products which seem most susceptible to change are first the
sugars and then the 70 per cent alcohol-soluble proteins, the 5 per
cent KjSOi-soluble protein and the water-soluble proteins
coagulated by so-called Stutzer's reagent.

"Com, barley, and oats are most subject to loss of sugar during
aging. On the other hand, many samples of wheat show a slight
loss the first year and then quite a rapid gain in the sugar content,
in some cases a gain 24 per cent of the total sugar present being
noted at the end of two years."

190 Original Communications: Eighth International [vol.

Dietary Studies and Dietetics

General dietary problems have been considered by many
writers with a view to making the work of the laboratory available
and useful to the housewife.

Methods of calculating the results of dietary studies and
similar topics are discussed in a publication of the American
School of Home Economics (Bui. Amer. School Home Econ.,
Ser. 1, 1909, No. 13, pi. 1, figs. 13), especially with reference
to the use of the so-called 100-calorie-portion-method of cal-
culating.

Emma S. Jacobs (Jour. Home Econ., 3 (1911), No. 2, pp.
162-168) in a discussion of family dietetics gives menus for what
she believes accurately planned dietaries for families, and a table
of weight and cost of protein and energy in different food mater-
ials designed to facilitate the computation of the nutritive value
of such dietaries.

Food customs and diet in American homes have been discussed
in a popular summary of data by C. F. Langworthy (U. S. Dept.
Agr., Office Expt. Stas. Circ. 110, pp. 32), which proposes dietary
standards for mineral constituents as well as for protein and
energy, and discusses dietary standards as distinguished from
physiological requirements.

Nellie M. Dickinson (III. Agr., 12 (1908), No. 5, pp. 142-145)
gives data regarding the preparation of a day's ration designed to
conform to dietary standards.

Information regarding dietary habits, food supply, and living
conditions of native tribes often appears in descriptive articles,
books of travel, reports of ethnological investigations, and other
publications not directly concerned with nutrition, which is of
importance in discussions of nutrition problems as well as of
general interest. As illustrations of such work the following
may be cited:

The food of natives of the upper Yukon has been described
by F. Schmitter (Smithsn. Misc. Collect., 56, No. 4, pp. 6, 7).
The diet of these natives consists of fish, game, and berries,
supplemented at the present time by vegetables bought at
local stores, though until recently they lived on animal food.

xvui] Congress of Applied Chemistry 191

The Mackenzie River natives, it is pointed out, live almost
exclusively on meat, and the author states that they are robust
and healthy.

A study of the food supply of Pima Indians has been reported
by F. Russell (Ann. Rpt. Bur. Amer. Ethnol., 26 (1904-5), pp.
66-92, figs. 7). These Indians subsist on a mixed diet in which
vegetable food predominates, buc it would seem probable that
the proportion of meat used was greater in the past than at
present.

Much information regarding the kind and amount of food used
by native Indian tribes in the southwestern United States and
northern Mexico is included in a physiological and medical study
of the Indians by A. Hrdlicka (Smithsn. Inst., Bur. Amer.
Ethnol. Bui. 34, pp. IX. + 460, pis. 28, figs. 2). Meat, corn,
some vegetables, and other foods make up a simple mixed
diet. Cooking processes are described as well as dietary habits
and customs.

In a volume on "Mexico" (New York and London, 1909, pp.
213-218, pi. 1) C. R. Enock reports data regarding the Mexican
peons, or country people. Corn meal, the native beans, fat,
and meat when it can be obtained, are the principle articles
of diet.

Of popular summaries which contain data of interest from the
standpoint of diet may be mentioned a paper by C. W. Furlong
(Harpers' Mo. Mag., 120 (1910), No. 716, pp. 217-229, pi. 1, figs.
7, map 1), which gives considerable information regarding the
character of the diet of the natives of Tierra del Fuego, which
consists almost entirely of the meat of wild animals, birds, the
blubber from stranded whale, fish, and mussels.

In a later paper by Furlong (Harpers' Mo. Mag., 122 (1911),
No. 732, pp. 813-827, pi. 1, figs. 9, maps 2) additional informa-
tion is given on the subjecc, parcicularly regarding the food cus-
toms and living conditions of the Tehuelches of the Patagonian
pampas. Apparently, these natives live very largely upon the
meat of mares and game.

Less work pertaining to the food consumption of fanulies
and groups has been reported ihan in some other branches of
dietetics.

192 Original Communications: Eighth International [vol.

From data regarding the food of a poor family in Buffalo and
one in Boston published by Emma 0. Lundberg (Survey 23,
(1910), No. 20, pp. 728-730) the protein and energy in the daily
food have been calculated.

As part of its nutrition investigations the Office of Experiment
Stations has reported the results of 4 dietary studies of farmers'
families in Vermont (J. L. Hills), 70 in mountain regions in Ten-
nessee (C. E. Wait), and 14 in Georgia (H. C. White). The cost
of nutrients and energy, peculiarities of the diet, adequacy of the
food supply, and similar questions are discussed, the results
being compared with earlier work of a similar nature. As a
whole the bulletin supplies a large amount of statistical and other
data regarding living conditions in rural regions, particularly
those remote from large centers of population where conditions
are very different from those which prevail in towns, cities, and
farms which are otherwise situated. (U. S. Dept. Agr., Office
Expt. Stas. Bui. 221, pp. 142, pis. 4).

The report of E. T. Wilson (Ann. Rpt. Isthmian Canal Com.,
1910, pp. 323-325), the subsistence officer in charge of the sub-
sistence department. Isthmian Canal Commission, contains
data regarding the kind and amount of food served to laborers
in the Panama Canal Zone. Using the average values for the
composition of foods as purchased, it has been calculated that
the European laborers' messes would supply 201 gm. protein
and 5,428 calories energy per person per day, and the common
laborers' kitchens 148 gm. protein and 4,680 calories energy.
The amounts actually eaten were not calculated, as no data
regarding the waste and refuse were available.

The scientific work organized under government auspices in
the Philippines has provided an opportunity for important stud-
ies of the native dietary. E. D. Merrill (Philippine Jour. Sci., B.
Med. Sci., 4 (1909), No. 4, pp. 219-223) has studied the principal
foods used by the natives of Taytay, and H. Aron (Philippine
Jour. Sci., B. Med. Sci., 4 (1909), No. 4, pp. 225-231) has studied
their food from a physiological standpoint. Rice is the staple
nonnitrogenous food and is supplemented by fish, fruits, and some
similar foods. The composition of a number of sorts of food was
determined.

xvin] Congress of Applied Chemistry 193

The matter has also been studied by V. G. Heiser (Ann.
Rpt. Bur. Health Philippine Islands, 1909, pp. 25-29), his
paper being entitled "Diet and Nutrition of the Filipino
People."

That the diet in public institutions may now be passed upon
by an expert in a manner profitable to the institution as well
as of interest to the investigator is one of the important, results
of the nutrition work of the last 25 years or more. This has been
obtained very largely as a result of the numerous investigations
of dietaries in general and public institutions dietaries in partic-
ulai, carried on as a part of the nutrition investigations of the
Office of Experiment Stations and related enterprises.

A report of work of this character issued recently by the De-
parcmeat of Agriculture contains the results of studies in a home
for old ladies and an orphan asylum in Philadelphia (Emma
Smedley and R. D. Mihier) and in orphan asylums, homes for
the aged, and a public home whose inmates are chiefly middle-
aged or aged people, in Baltimore (H. L. Knight, H. A. Pratt,
and C. F. Langworthy). On the basis of the data reported the
dietaries are critically considered and some changes suggested.
The general problem of the dietary of children and the dietary
of aged persons is discussed at length particularly with reference
to pubHc institutions. (U. S. Dept. Agr., OflSce Expt. Stas.
Bui. 223, pp. 98).

Considerable information is given regarding the character of
the diet in a state hospital for the insane in Illinois in connection
with a study of the occurrence of pellagra at the institution,
carried on by J. F. Siler and H. J. Nichols (111. Bd. Health Mo.
Bui., 5 (1909), No. 7, pp. 437-478, figs. 8). On an average the
simple mixed diet supplied approximately 30 gm. of protein and
2,000 to 2,500 calories per day.

In connection with an exhaustive and important investigation
of the methods of fiscal control of state institutions carried on
for the Sage Foundation, H. C. Wright (State Charities Aid
Assoc. [N. Y.] Pub. 122, 1911, pp. 353) reports the calculated
food suppUed per man per year and per man per day in insti-
tutions in New York, Indiana, and Iowa, including hospitals for
the insane, soldiers' homes, industrial schools, reformatories.

194 Original Communications: Eighth International [vol.

prisons, and institutions for the feeble-minded and for epilep-
tics. The results are summarized as follows:

Average Food peb Man peb Yeab and NtrTRiTivE Value of Dailt Ration
IN Public Institutions

Location of institutions

Food per man per
year

Food per man per
day-

Total
amount

Cost

Protein

Energy

New York

Pounds
1,227
1,176
1,423

$45.05
43.03
55.48

Grams

104.62

98.78

106.47

Calories
3,313
3,429
3,691

InfliaTia

Iowa

Numerous investigations of the food and diet of children have
been made during the period under consideration. Though all
that pertains to food from infancy to maturity is obviously of
interest and value, special studies of infant feeding have not been
included in this summary, since they are commonly regarded as
pertaining to the subject of medicine rather than to dietetics.

The bearing of food during early life upon the normal develop-
ment of the body is a question which has received attention from
a number of investigators, the problem having been studied with
young animals as well as with children.

In an investigation of this sort experimental studies with dogs
were reported by H. Aron, together with the results of general
observations on nursing children (Philippine Jour. Sci., B. Med.
Sci., 6 (1911), No. 1, pp. 1-52, pis. 4, dgms. 5).

He concludes that "a growing animal which receives only
sufficient food to keep its body weight constant, or to allow
slight increase, is in a condition of severe starvation. If by a
restriction of food the increase in weight is inhibited, the skeleton
grows at the expense of other parts of the body, expecially of the
flesh. Most of the organs retain their weight and size, while the
brain grows to reach its normal weight. The composition of the
body — when at a constant weight — undergoes remarkable

xviii] Congress of Applied Chemistry 195

changes. Fat is consumed more or less entirely, the quantity
of protein, especially of the muscles but not of the organs, is
diminished and a great proportion of the body tissues is replaced
by water; thus, this water and the increase of the skeleton
together replace the body materials lost. The caloric value of
1 gm. body weight of an animal which has undergone such a
process to its extreme limit may amount to only one-third of the
normal value.

"It is possible by supplying suitable amounts of food to main-
tain a dog in an emaciated condition, apparently in good health,
and at the weight of a puppy, for nearly 1 year, while its weight
at the end of the year should be 3 times as greac. If such an
animal is thereupon fed amply, it fattens and rounds out, but
does not reach the size of a control animal which from the begin-
ning has been normally fed. It is unable to make good the growth
suspended by the long restriction of food.

"The 'growth' principally depends on the tendency to grow
possessed by the skeleton. The skeleton loses its capability
of growing in more advanced age regardless of the size which the
animal has reached."

A paper of importance in considering the question of physio-
logical requirement and dietary standard has been published by
H. J. Waters (Proc. Soc. Prom. Agr. Sci., 30 (1909), pp. 70-98;
Separate, pp. 29, figs. 6), who discusses the influence of nutrition
upon the animal form, as the result of a large number of experi-
ments made with farm animals (beef cattle). In general, the
results clearly show that a decreased supply of nourishment in
the young animals hinders body development. The matter is
also taken up in a later paper (Quart. Rpt. Kans. Bd. Agr., 29
(1910), No. 113, pp. 59-86, figs. 7).

The relationship of food to physical development is discussed
by D. McCay (Philippine Jour. Sci., B. Med. Sci., 5 (1910), No.
2, pp. 163-170), on the basis of his investigations, his conclu-
sion being that there is a close relationship between the nutritive
value of the diet, and particularly its protein content, and physi-
cal development. This he believes is clearly brought out in a
comparison of the degree of nitrogenous interchanges of a num-
ber of native races, arranged according to the amount of nitro-

196 Original Communications: Eighth International {vol.

gen per kilogram of body weight. At the head of the list as
regards physical development are the Nepalese Bhutias, with
0.42 gm., and the Tibetan and Bhotan Bhutias, with 0.35 gm.,
respectively, of nitrogen per kilogram of body weight and with
a large amount of animal food in the diet, and at the bottom of
the list are the Bengalis and Ooriyas, with 0.116 gm. of nitrogea
per kilogram of body weight.

Few dietary studies with children have been reported in the
United States during the period under consideration.

The general question of the feeding of yoimg children has been
discussed in a popular way by Mary Swartz Rose (Teachers
Col. [N. Y.] BuL, 2. ser., 1911, No. 10, pp. 10) and by others.

The dietary studies made in orphan asylums by the Depart-
ment of Agriculture (U. S. Dept. Agr., Office Expt. Stas. Bui.
223) have been referred to elsewhere. The report of this work
contains a discussion of children's dietaries and proposed dietary
standards.

An extended summary of data regarding the daily meals of
school children was prepared by Caroline L. Hunt (Bur. of Ed.
[U. S.] Bui. 3, 1909, pp. 62, pis. 3, dgm. 1) for the U. S. Bureau
of Education. The paper is an important contribution to the
subject, not only from the material it brings together, but also
because of the suggestions it makes regarding the rational feedmg
of school children.

School luncheons have also been studied under the auspices of
the Women's Educational and Industrial Union (Ann. Rpt.
"Women's Ed. and Indus. Union, 29 (1908), pp. 34, 35).

Lillian D. Wahl (Charities and Commons, 20 (1908), No. 11,
pp. 371-374) has reported data on this subject, particularly
regarding attempts made to supply food lo children in some of
the New York public schools.

Contributions to the school luncheon problem are also made
by Marion Bell (Boston Cooking-School Mag., 12 (1908), No.
6, pp. 292, 293) in an article describing a luncheon cooked and
served in the Honolulu Normal School.

An important contribution to the general question is the study
of malnutrition of children in New York public schools made by
E. M. Sill (Jour. Amer. Med. Assoc, 52 (1909), No. 25, pp. 1981-

xviii] Congress of Applied Chemistry 197

1985). He found that 83 per cent of the 210 cases observed
depended practically for their diet on bread with tea or coffee.
The importance of a highly nutritious dietary, with a large amount
of protein, is recognized, and suggestions given regarding the
preparation of such a diet.

Continuing his work regarding the diet of undernourished
school children in New York City (Jour. Amer. Med. Assoc, 55
(1910), No. 22, pp. 1886-1891), Sill studied the dietary of 28
families with malnourished children in the thickly congested
districts of New York City and of 6 families in more comfortable
circumstances but where the children were also undernourished.

In the first group the food cost 19 cents per man per day, and
supplied 95 gm. protein, 68 gm. fat, and 407 gm. carbohydrates,
the fuel value being 2,614 calories. The families were engaged
in active or moderately active muscular work.

In the fairly well-to-do families the diet cost on an average
35 cts. per person per day, and supplied 149 gm. protein, 115 gm.
fat, and 569 gm. carbohydrates, the fuel value being 3,884
calories. The families were engaged in moderately active work.

Where the dietaries were up to or above the standard the
malnutrition was attributed to such factors as close quarters,
over crowding, eating candy between meals, tuberculous infec-
tion, enlarged tonsils, or other similar cause. The author states
that in his experience such childien contract disease much more
easily and have less resistance than well-nourished children.

Information regarding undernourished children in New York
City was collected by Frances Perkins (Survey, 25 (1910), No. 1,
pp. 68-72). According to her summary, "physical disabilities of
one kind or another are closely associated with malnutrition,
and make it doubly dangerous."

W. C. HoUopeter (Jour. Amer. Med. Assoc, 53 (1909), No. 21,
pp. 1727-1730), who has studied the character of breakfasts of
over 2,000 school children, has also contributed important infor-
mation concerning existing conditions with reference to chil-
dren's diet.

Numerous plans for providing proper food for school children
have been proposed, particularly with reference to needy or
undernourished children.

198 Original Communications: Eighth International [vol.

A 'luncheon project which has proved successful in Philadel-
phia is described by H. H. Bonnell (Starr Center Assoc. [Rpt.j
1909, pp. 18-20, fig. 1), the different articles being sold for a
penny.

The subject of penny luncheons for school children in the
thickly congested districts of Philadelphia is discussed further
by Alice C. Boughton (Psych. Clin., 3 (1910), No. 8, pp. 228-231,
fig. 1).

Data regarding the serving of penny lunches to school chil-
dren in Boston are reported by Ellen H. Richards (Jour. Home
Econ., 2 (1910), No. 6, pp. 648-653).

A school luncheon costing one cent per person, which fur-
nishes, in round numbers, 9 gm. of protein, is described by A.
L. Benedict (Dietet. and Hyg. Gaz., 23 (1907), No. 7, p. 404).

Some work has been done with older students and pupils.

Daily menus for the school year are presented, together with
the results of a dietary study for 1 month, in a report issued
by the Institute for Colored Youth (Teachers' Training School)
(Cheyney, Pa., 1909, pp. 48). "the work was done as a part
of a project to prepare at reasonable cost a rational diet with
protein and energy in accordance with commonly accepted
dietary standards.

Agnes Hunt (111. Agr., 12 (1908), No. 5, pp. 146-148) reports
data regarding the nutritive value and cost of food served in
a students' boarding-house.

From data reported by P. R. Kellar (Cooking Club Mag.,
12 (1910), No. 11, pp. 10, 11), regarding the diet in a students'
boarding house in the University of Minnesota, the daily food
which cost 22 cts. per man per day was calculated to supply
105 gm. protein and 3,715 calories of energy.

Of special investigations of dietary problems the following
may be cited.

The possible relation of diet to fatigue, particularly a diet
containing the usual average amount of protein, is discussed
by I. Fisher (Bui. Com. One Hundred Nat. Health [Wash-
ington], 1909, No. 30, pp. VIIH-138) in a report on national
vitality, its wastes, and conservation. The author is of the
opinion, from experiments which he has made, that there is a

xviii] Congress of Applied Chemistry 199

relationship between protein consumption and the occurrence
of fatigue.
Studies carried on by P. A. Shaffer (Amer. Jour. Physiol.,

22 (1908), No. 4, pp. 445-455) with healthy men support the
belief that with sufficient food either an increase or a decrease
of muscular activity within physiological limits has per se no
effect upon protein metabolism, as indicated by the nitrogen
and sulphur partitions in the urine. The investigation as a
whole was undertaken to secure data regarding diminished
muscular activity and protein requirement.

The effect of an ash-free diet was studied experimentally by
H. W. Goodall and E. P. Joslin (Trans. Assoc. Amer. Physicians,

23 (1908), pp. 92-106) with healthy individuals, for experi-
mental periods of 13 and 9 days, respectively. The results
obtained do not indicate any marked changes in metabolism
ascribable to the ash-free diet.

A contest entered into for a wager, in which 48 men endeav-
ored to carry on the back a weight of 100 lbs., for approximately
10 miles, furnished results which were discussed with reference
to the strength and endurance of the men by C. F. Langworthy
(Science, n. ser., 33 (1911), No. 853, pp. 708-711). Of the
number who entered the contest 6 completed the task, while
the others dropped out at various stages. The information col-
lected from a number of the men showed that they lived on a
simple mixed diet. The energy expended in moving the body and
carrying the load over the course was calculated to be 1,137
calories on an average for the 6 successful contestants, of which
amount 707 calories would represent the energy expenditure for
motion of forward progression and 430 calories for energy ex-
pended in moving the load.

Similar calculations for individuals and for groups are reported.

It seems fair to conclude that the men who engaged in the
contest were, as regards their food, their occupation, and their
general living conditions, representative of a very large group
of our population who are living comfortably and meeting their
daily obligations in a creditable manner, who are, in fact, living
the average life of the average man, with its varied activities
and interests.

200 Original Communications: Eighth International [vol.

In so far as the recorded data throw light on the subject, they
indicate that the average man living the average life is capable
of meeting body demands of considerable severity — a conclu-
sion which perhaps few would question, but which it is inter-
esting to consider in the light of numerical data.

Digestion

Studies of thoroughness of digestion have been reported as
has work on various details of the general question. The ten-
dency seems to be toward the investigation of special topics
rather than the digestion as a whole. Experiments have also
been made with reference to ease of digestion, which involve
the use of the respiration calorimeter. (See p. 73).

Colloid-chemical aspects of digestion are considered by J.
Alexander (Jour. Amer. Chem. Soc, 32 (1910), No. 5, pp. 680-
687), who reports ultramicroscopic observations. The author's
general conclusion is that chemical analysis alone is not sufficient
to express the digestibility and availability of food and that our
consideration of such problems must be broadened.

The absorption of fat was studied by L. B. Mendel (Amer.
Jour. Physiol., 24 (1909), No. 5, pp. 493-496) who used samples
stained with Sudan III. He concludes that "when fat stained
with water-soluble dyes, like Sudan III, is fed, the pigments
readily pass into the lymphatic vessels and thereby reach the
blood stream. Since these compounds are soluble in free fatty
acids as well as in neutral fats, their presence in the lymph can-
not be taken as evidence either for or against the possibiUty of
the digestion of fats prior to their absorption."

The effect of the presence of carbohydrates upon the artificial
digestion of casein was studied by Nellie E. Goldthwaite (Jour.
Biol. Chem., 7 (1910), No. 2, pp. 69-81), the recorded data
showing, according to the author, that each of the carbohy-
drates tested (glucose, maltose, dextrose, dextrin, and galactose)
retarded the digestion of casein, the retardation being propor-
tional to the amount of added carbohydrate.

The results of experiments by C. H. Neilson and D. H. Lewis
(Jour. Biol. Chem., 4 (1908), No. 6, pp. 501-506, fig. 1) on the

xviii] Congress of Applied Chemistry 201

effect of diet on the amyloljrfcic power of saliva led them to
conclude that "there is a change either in the amount of ptyalin
or in its activity, or in the concentration of the saliva, which
enables more or less starch to be digested with a given quantity
of saliva according to the diet. . . . Whether this change
in the amylolytic power of the saUva due to diet should really
be called an adaptation to diet is immaterial."

G. Lusk (Amer. Jour. Physiol., 27 (1911), No. 5, pp. 467, 468)
has summarized data accumulated in connection with some of
his earlier experiments, which have to do with the questions as
to whether dextrose arises from cellulose in digestion, the con-
clusion being that such is not the case.

According to the conclusions reached by J. H. Pratt and L.
H. Spooner (Trans. Assoc. Amer. Physicians, 25 (1910), pp.
614-635) in a study of the internal functions of the pancreas
in carbohydrate metabolism, there is a rapid decrease in the
power to assimilate glucose after the onset of atrophy of the
pancreas.

Using dogs with Pawlow fistules, N. B. Foster and A. V. S.
Lambert (Proc. Soc. Expt. Biol, and Med., 4 (1906), No. 1,
p. 13) studied the influence of water on gastric secretion and the
chemical affinity of mucus for hydrochloric acid in the stomach.

"It was observed that with definite amounts of cracker meal
as food, the amount and rate of gastric secretion depend to
some extent on the amount of water given the dog with his
meal, i.e., when small amounts of water are given, the secretion
is slow and scanty. If larger quantities of water are mixed in
the food the secretion is more abundant.

"The degree of acidity of gastric juice depends upon the
amount of secretion. When this is considerable it is much more
acid than when the secretion is scanty. . . . The proportion
of free acid depends upon the amount of mucus secreted, since
mucus protein Hke other proteins combines with HCl. Mucus
in the presence of pepsin combines with HCl to a considerable
extent and undergoes digestion, with formation of proteoses."

The results of a large number of experiments carried on by
the Office of Experiment Stations cooperating with the Bureau
of Animal Industry on the digestibility of cheese of different

202 Original Communications: Eighth International [vol.

sorts have been reported in a summary prepared by C. F. Doane
(U. S. Dept. Agr., Bur. Anim. Indus. Circ. 166, pp. 22). In
the first series there were 184 tests with 65 young men serving
as subjects, and in the second series about 50 experiments.
American cheese made by the regular Cheddar process, with
varying amounts of rennet and cured for different lengths of
time and ripened under controlled conditions, was used in the
tests, as well as a number of other sorts of cheese, with a view to
determining whether thoroughness of digestion was influenced by
the kind of cheese, by the degree of ripeness, and by similar factors.

In general, cheese of all sorts was found to be very thoroughly
digested and little or no difference in the comparative digesti-
bility of cheese at different stagey of ripeness was observed.
It was also found that different kinds of cheese closely resem-
bled cheese made by the Cheddar process in thoroughness of
digestion, and, in general, "that all kinds of cheese, even the
very high-flavored and so-called condimental cheeses, have a
high food value."

Brief statements are also made regarding the experiments on
the ease of digestibility of cheese in which the respiration
calorimeter was used.

The digestibility of white of egg as influenced by the tem-
perature at which it is coagulated was studied by P. Frank
(Jour. Biol. Chem., 9 (1911), No. 6, pp. 463-470, dgms.2).
The progress of the hydrochloric acid action and the total
digestion is most rapid in the albumin not heated beyond 75°C.

The results are reported of 66 natural and 99 artificial digestion
experiments with meat undertaken by H. S. Grindley, T. Mo-
jonnier, and H. C. Porter (U. S. Dept. Agr., Office Expt. Stas.
Bui. 193, pp. 100), to determine the ease and thoroughness of
digestion of different kinds and cuts of meat cooked in a variety
of ways. As regards thoroughness of digestion, the results do
not indicate that very appreciable differences exist, and meats
of all kinds and cuts can be classed with the very digestible
foods, about 98 per cent of the protein and fat being retained
in the body on an average.

The erepsin of cabbage has been studied by Alice F. Blood
(Jour. Biol. Chem., 8 (1910), No. 3, pp. 215-225). It splits

xviii] Congress of Applied Chemistry 203

tryptophan from Witte's peptone and casein, and tyrosin from
peptone "Roche." It clots milk and liquefies gelatin. It does
not digest fibrin, coagulated egg white, or edestin in neutral,
acid, or alkaline solution, or in the presence of HCN. Other
characteristics are given.

An extended study of the digestibility and nutritive value of
legumes was carried on by C. E. Wait (U. S. Dept. Agr., OflBce
Expt. Stas. Bui. 187, pp. 55), of the University of Tennessee,
in connection with the nutrition investigations of the Office of
Experiment Stations. In general, kidney beans, white beans,
and cowpeas of different sorts were found to supply from 70 to
83 per cent digestible protein and from 87 to 96 per cent di-
gestible carbohydrates. When the digestibility of a diet con-
taining a considerable quantity of beans was considered, rather
than that of the beans alone, the values were higher. The
experiments as a whole demonstrate the great nutritive value
of cowpeas.

A number of foods, particularly tropical fruits and vege-
tables, were analyzed by L. H. Merrill (Maine Sta. Bui. 158,
pp. 219-238), of the Maine Experiment Station, and digestion
experiments with hulled corn reported. These show in general
that the digestibility of protein and the availability of energy
are low in comparison with results obtained with white bread.
Analysis of com before and after popping showed that except
for loss of water little change in composition was produced by
this process.

Studies of digestibility of carbohydrates of marine algae by
Mary D. Swartz have been referred to elsewhere.

Metabolism; Respiration Calorimeters, Bomb Calori-
meters, AND Experiments with Them

A fairly large proportion of the work reported during the
period under consideration has had to do with the metabolism
of nitrogen and other constituents, including numerous studies
of the metabolism of the income and outgo of energy made
with the respiration calorimeter, and with special studies of
body excretions, the influence of different food constituents

204 Original Communications: Eighth International [vol.

upon protein consumption, the effects of water drinking, and
similar topics.

The average daily excretion of uric acid of 10 men m normal
condition, ranging from 19 to 29 years, and fed a normal diet,
was determined by P. J. Hanzlik and P. B. Hawk (Proc. Soc.
Expt. Biol, and Med. 6 (1908), No. 1, pp. 18, 19, and found
to be 0.597 gm., a value somewhat lower than the generally
accepted average of 0.7 gm.

"The average daily protein ingestion from these same subjects,
when permitted to select their diet, was 91.2 gm. or 1.33 gm. for
such a period."

The metabolism of some purin compounds in the rabbit, dog,
pig, and man was studied by L. B. Mendel and J. F. Lyman
(Jour. Biol. Chem., 8 (1910), No. 2, pp. 115-143).

In the experiments with man hypoxanthin nitrate, xanthin,
guanin, and adenin were added on different days to a purin-free
diet. According to the authors, the examination of the urine
showed that all four purins produced a marked rise in urinary
acid and a small, yet noticeable increase in the elimination of
purin bases. The smaller and lighter of the two subjects ex-
creted, in every case, a larger percentage of uric acid and
purin bases than the other subject, and, according to the
authors, "may possess a more limited power for uric acid
destruction."

The effect of meat purins (largely free hypoxanthin), on the
elimination of purin compounds is illustrated by data cited
from a series of experiments by Hilditch, also made at Yale
University, in which meat was substituted for the milk and
eggs of a purin-free diet. The resulting increase in the excretion
of uric acid nitrogen, it is stated, is quite comparable with the
figures obtained in the experiment with pure hypoxanthin.

In discussing their work in comparison with that of earlier
investigators, the authors point out that the data which they
report "emphasize the fact that all of the familiar purins may
lead to an increase in exogenous uric acid in the urine of man,
with (quantitatively) little influence on the elimination of the
purin bases. They may be interpreted to support the most
prevalent view that uric acid is a stage in the metabolism of

xviii] Congress of Applied Chemistry 205

exogenous purins in the human body, a view rendered especially
plausible by the growing statistics on tissue enzyms."

The composition of dilute renal excretions was studied by A.
B. Macallum and C. C. Benson (Jour. Biol. Chem., 6 (1909), No. 2
pp. 87-104), the general conclusion being that the elimination
of water, potassium salts, and chloride from the body is not
due to filtration, but in the case of water, to the physiological
activity of the renal membranes involved, and in the case of the
salts, to forces which may be termed "secretory."

T. B. Barringer, Jr., and B. S. Barringer (Amer. Jour. Phy-
siol, 27 (1910), No. 1, pp. 119-121) have compared the total
nitrogen excretion of either kidney in normal individuals during
varying periods of time. In one case the excretions from the
2 kidneys were found to be equal in quantity. Six times they
varied by less than 10 per cent and 4 times from 10 to 20 per
cent.

"As regards the total nitrogen, in one case the quanitites were
equal. In 7 cases they varied by less than 1 gm. per liter. In
2 cases they varied by between 1 and 2 gm. per liter. The
nitrogen-urea plus ammonia-urea showed in 3 cases a variation
less than 1 gm. per liter and in 6 cases a variation of between 1
and 2 gm."

L. W. Riggs (Abs. in Jour. Biol. Chem., 9 (1911), No. 2, p.
XIX; Proc. Amer. Soc. Biol. Chem., 2 (1910), No. 1, p. 13)
studied the chemical composition of human sweat, using 45 sam-
ples obtained from persons in normal health and from nephritics.
The total nitrogen, nitrogen as urea plus ammonia, inorganic
acids, potassium, and chlorin were determined in the majority of
the samples.

Factors regulating the creatinin output in man were studied
by P. A. Levene and L. Kristeller (Amer. Jour. Physiol., 24 (1909),
No. 1, pp. 45-65), the experimental data apparently indicating
that the formation of creatin and creatinin represents two phases
in the catabolism of a single substance. The constant activity of
the creatinin output in normal men, the authors believe, is con-
ditioned by the high velocity of creatin combustion in health.

The process of acid excretion was studied critically by L. J.
Henderson (Jour. Biol. Chem., 9 (1911), No. 5, pp. 403-424, dgms.

206 Original Communications: Eighth International [vol.

3), who concludes that, as is the case with temperature and
osmotic pressure, normal neutrality or alkalinity is adjusted by a
mechanism within the body, but is maintained permanently by
exchanges with the environment.

In a study of the nutritive value of gelatin, J. R. Murlin (Amer.
Jour. Physiol., 19 (1907), No. 3, pp. 285-313) found that under
certain conditions, namely, supplying a large proportion of the
energy of the ration in the form of carbohydrates being especially
favorable, it was possible in experiments with man and dogs to
replace part of the proteid nitrogen with gelatin nitrogen for
maintaining nitrogen equilibrium at a fasting level.

Gelatin was one of the materials included in a study of the
elimination of total nitrogen, urea, and ammonia following the
administration of some amino acids, glycylglycin and glycylglycin
anhydrid by P. A. Levene and G. M. Meyer (Amer. Jour. Physiol.,
25(1909), No. 4, pp. 214-230). The experiments were made with
dogs. According to the authors, all of the excessive nitrogen
added as gelatin to a standard diet "is eliminated in the form of
urea. Thus, this experiment leads to the conclusion that either
diketopiperazins do not enter into the composition of the protein
molecule, or that the anhydrids of peptids within the protein
molecule offer less resistance than when in a free state."

The significance of glycocol and carbohydrate in sparing the
body's proteid was also studied by J. R. Murlin (Amer. Jour.
Physiol., 20 (1907), No. 1, pp. 234-258). A specific relationship
was shown to exist between carbohydrates ingested and the
elimination of nitrogen, carbohydrate not needed for combustion
being far more efficient for reducing nitrogen output than car-
bohydrate coming within the requirement for potential energy.
This fact, according to the author, indicates the importance of
abundant carbohydrates for convalescence and growth, and may
explain the almost universal craving for sweets especially in the
young.

From experiments with animals on mucic acid and carbohydrate
metabolism L. B. Mendel and W. C. Rose (Abs. in Jour. Biol.,
Chem., 9(1911), No. 2, p. XII; Proc. Amer. Soc. Biol. Chem.,
2 (1910), No. 1, p. 6) conclude that this acid "is presumably not
an intermediary oxidative product in the metabolism of galactose

xnn] Congress of Applied Chemistry 207

or galactose-yielding carbohydrates. The urinary oxalic acid is
only very slightly increased after the ingestion of large amounts
of mucic acid. This increase is by no means as large as would be
expected if mucic acid were a precursor of oxalic acid."

Some experiments on the influence of caffein on protein meta-
bolism of dogs have been reported by W. Salant and I. K. Phelps
(Jour. Pharmacol, and Expt. Ther., 2 (1911), No. 4, pp. 401, 402),
who also discuss demethylation in the body. The resistance to
caffein, the authors state, "was found to vary with the amounts
of the urinary purins eliminated."

W. Salant and J. B. Rieger (Jour. 'Pharmacol, and Expt. Ther.,
2 (1911), No. 4, pp. 400, 401) studied the elimination of creatin
and creatinin after the administration of caffein, with rabbits.
The results indicate that urinary creatin is increased after the
administration of caffein, the size of the dose being an important
factor. Neither the increased diuresis nor the diminished
appetite observed could, in the authors' opinion, be regarded as a
factor in accounting for the increased output of creatin.

On theoretical grounds, G. Lusk (Zentbl. Physiol., 21 (1907),
No. 26, pp. 861, 862) discusses the specific dynamic effect of por-
tein.

According to H. McGuigan's (Jour. Biol. Chem., 3 (1907),
No. 3, Proc, pp. XXXVII, XXXVIII) studies of sugar meta-
bolism in vitro, the clinical assertion is maintained that levulose,
is more easily oxidized than glucose and that it may be used in
the body when glucose can not. The order of ease of oxidation of
a number of sugars is as follows: Levulose, galactose, glucose,
maltose, and saccharose, levulose being the most easily oxidized.

H. McGuigan (Amer. Jour. Physiol., 21 (1908), No. 3, pp.
334-350) found that the living muscles of an animal, when per-
fused with dextrose, levulose, or galactose, caused a rapid oxida-
tion of these sugars. With maltose direct oxidation was not
noted. Other questions were also considered in this experimental
inquiry, which is a contribution to the question of the way in
which the animal body utilizes a carbohydrate food supply.

The income and outgo of nitrogen of a simple mixed diet are
determined and briefly reported by Clara C. Benson et al. (Jour.
Home Econ., 2 (1910), No. 6, p. 658).

208 Original Communications: Eighth International [vol.

An extended series of studies with fasting subjects has been
reported by P. B. Hawk and his associates, in which the following
matters have been taken up : Nitrogen partition and physiological
resistance as influenced by repeated fasting (Jour. Amer. Chem.
Soc, 33 (1911), No. 2, pp. 215-254, dgm. 1); the catalase content
of tissues and organs after prolonged fasting (Jour. Amer. Chem.
Soc, 33 (1911), No. 3, pp. 425-434) ; the nitrogen partition of two
men through seven-day fasts following the prolonged ingestion of
a low protein diet, supplemented by comparative data from the
subsequent feeding period (Jour. Amer. Chem. Soc, 33 (1911),
No. 4, pp. 568-598); the allantoin and purin excretion of fasting
dogs (Jour. Amer. Chem. Soc, 33 (1911), No. 10, pp. 1601-1622);
the influence of an excessive water ingestion on a dog after a pro-
longed fast (Jour. Biol. Chem., 10 (1911), No. 5, pp. 417-432);
distribution of nitrogen during a fast of one hundred and seven-
teen days (Jour. Biol. Chem., 11 (1912), No. 2, pp. 103-127, dgm.
1); the putrefaction processes in the intestine of a man during
fasting and during subsequent periods of low and high protein
ingestion (Jour. Biol. Chem., 11 (1912), No. 3, pp. 169-177);
hydrogen ion concentration of feces (Jour. Biol. Chem., 10 (1912),
No. 2, pp. 129-140); on the differential leucocyte count during
prolonged fasting (Amer. Jour. Physiol., 30 (1912), No. 2, pp.
174-181); and glycogen-free liver (Jour. Amer, Chem. Soc, 34
(1912), No. 6, pp. 826-828).

Hawk and his associates have also reported an extended series
of experiments on the relative effects of copious and moderate
water drinking with meals, some of the experiments being carried
on as a part of the series of fasting tests referred to above. The
following list of subjects studied show the character and extent of
the work: The influence of copious water drinking (Univ. Penn.
Med. Bui., 18 (1905), No. 1, pp. 7-25); the stimulation of gastric
secretion under the influence of water drinking with meals
(Jour. Biol. Chem., 9 (1911), No. 2, pp. XXIX, XXX; Proc.
Amer. Soc. Biol. Chem., 2 (1910), No. 1, pp. 23, 24); the meta-
bolic influence of copious water drinking with meals (Jour. Expt.
Med., 12 (1910), No. 3, pp. 388-410); the uric acid elimination
following copious water drinking between meals (Jour. Amer.
Chem. Soc, 32 (1910), No. 12, pp. 1686-1691); the excretion of

xviii] Congress of Applied Chemistry 209

chlorids following copious water drinking between meals (Re-
printed from Arch. Int. Med., 7 (1911), pp. 536-550); intestinal
putrefaction during copious and moderate water drinking with
meals (Arch. Int. Med., 7 (1911), No. 5, pp. 610-623); the activity
of the pancreatic function under the influence of copious water
drinking with meals (Arch. Int. Med., 8 (1911), pp. 382-394);
the allantoin and purin excretion of fasting dogs (Jour. Amer.
Chem. Soc, 33 (1911), No. 10, pp. 1601-1622); the utilization
of ingested fat under the influence of copious and moderate water
drinking with meals (Jour. Amer. Chem. Soc, 33 (1911), No. 12,
pp. 1978-1998); the distribution of bacterial and other forms of
fecal nitrogen and the utilization of ingested protein under the
influence of copious and moderate water drinking with meals
(Jour. Amer. Chem. Soc, 33 (1911), No. 12, pp 1999-2019), a
fecal output and its carbohydrate content under the influence
of copious and moderate water drinking with meals (Jour. Amer.
Chem. Soc, 33 (1911), No. 12, pp. 2019-2032); the influence of
an excessive water ingestion on a dog after a prolonged fast
(Jour. Biol. Chem., 10 (1911), No. 5, pp. 417-432); the allantoin
output of man as influenced by water ingestion (Jour. Amer.
Chem. Soc, 34 (1912), No. 4, pp. 546-550); and the hydrogen ion
concentration of feces (Jour. Biol. Chem., 11 (1912), No. 2, pp.
129-140).

As a whole the investigations, which are too extended to be
quoted in detail, were favorable to the use of water with meals.

The metabolism of inorganic and organic phosphorus was
studied with laboratory animals (rabbits) by F. C. Cook (U. S.
Dept. Agr., Bur. Chem. Bui. 123, pp. 63, pis. 3), of the Bureau
of Chemistry. The rabbits fed organic phosphorus eliminated a
smaller proportion of ingested phosphoric acid in the lu-ine than
those fed inoiganic phosphorus, and the average amount of cal-
cium absorbed from the intestinal tract was higher in the case of
the rabbits fed organic phosphorus, the results agreeing with the
theory that calcium and phosphorus fed in inorganic form unite
to form insoluble calcium phosphate which is eliminated by the
bowels in ingested form. The amount of metabolized magnes-
ium that was retained indicates that the rabbits fed inorganic
phosphorus, while metabolizing a smaller amount of magnesium

210 Original Communications: Eighth International [vol.

than did those fed organic phosphorus, retained a larger per-
centage of the amount actually metabolized.

The sulphur balance in metabolism was studied by A. E.
Taylor (Abs. in Jour. Biol. Chem., 9 (1911), No. 2, pp. IX, X;
Proc. Amer. Soc. Biol. Chem., 2 (1910), No. 1, pp. 3, 4) with 6
normal men, for periods of nearly 3 months. A condition of
equilibrium was not observed, the output being regularly and
notably higher than the intake. The author does not consider
that the results obtained are trustworthy, the presumption being
that errors were involved in the determinations of the sulphur
index.

According to data reported by A. 0. Shaklee and S. J. Meltzer'
(Amer. Jour. Physiol., 25 (1909), No. 3, pp. 81-112), shaking
may completely destroy the three proteolytic ferments— pepsin,
rennin, and trypsin. "They are destroyed more rapidly at
higher than lower at temperatures; . . . trypsin is more
easily destroyed than pepsin; . . . shaking produced by the
respiratory movements is capable of causing some destruction
of the ferments. Recent experiments by other investigators show
also that other ferments may be inactivated by shaking . . .

"The assumption is here made that the nature of the destruc-
tion of ferments is similar to that which takes place in the destruc-
tion of living cells, and that shaking affects a certain structure
which is common to living cells as well as to red blood corpuscles
and to ferments."

In experiments on the effects of respiratory movements,
ferments in rubber or glass containers of suitable construction
were introduced into the stomach and peritoneal cavity, a dog
and rabbits serving as subjects.

E. W Rockwood (Proc. Iowa Acad. Sci., 15 (1908, pp. 99-103)
has studied the nature of the uric acid ferments which it is believed
are concerned in the formation of uric acid from nucleins in the
liver.

The problem of nuclein syntheses in the animal body was
studied experimentally by E. V. McCollum (Wisconsin Sta.
Research Bui. 8, pp. 75-93; Amer. Jour. Phisiol., 25 (1909), No.
3, pp. 120-141) with young and old rats, with normal and special
rations, such materials as edestin, zein, glucose, purified butter

xviii] Congress of Applied Chemistry 211

fat, and cane sugar being used. These purin-free diets and diets
containing purin bases were also compared.

Some of the author's conclusions were as follows:

"The palatability of the ration is a most important factor in
animal nutrition. Without palatability the ration may possess
all the necessary food ingredients and yet fail to nourish an
animal properly. . . .

"Very young animals adapt themselves to a ration possessing
a low degree of palatability much better than do adults.

"Other things being satisfactory, all the phosphorus needed
by an animal for skeleton, nuclein or phosphatid formation, can
be drawn from inorganic phosphates.

"The animal has the power to synthesize the purin bases neces-
sary for its nuclein formation from some complexes contained in
the protein molecule, and does not necessarily use purin bases of
exogenous origin for this purpose."

A useful summary and digest of data on the elementary compo-
sition of nuccleic acids, their constituents, and related questions
has been published by P. A. Levene (Jour. Amer. Chem. Soc,
32 (1910), No. 2, pp. 231-240), and a similar compilation on
oxidases by J. H. Kastle (Pub. Health and Mar. Hosp. Serv.
U. S., Hyg. Lab. Bui. 59, pp. 164).

Methods and standards in bomb calorimetry have been dis-
cussed on the basis of experience by J. A. Fries (U. S. Dept. Agr.
Bur. Anim. Indus. Bui. 124, pp. 32; Pennsylvania Sta. Rpt. 1909,
pp. 321-345).

An adiabatic calorimeter for use with the calorimetric bomb
has been described by F. G. Benedict and H. L. Higgins (Jour.
Amer. Chem. Soc, 32 (1910), No. 4, pp. 461-467, fig. 1).

The metabolism as a statistical problem has been considered by
H. L. Rietz and H. H. Mitchell (Jour. Biol. Chem., 8 (1910), No.
4, pp. 297-326). Such questions as the application of the laws
of probability, together with the various mathematical methods
of reducing statistical data, the importance of such procedure in
metabohsm experiments, and related questions are considered.

Comparative physiology of purin metabolism, by H. G. Wells
(Trans. Chicago Path. Soc, 7 (1909), No. 8, pp. 244-248; Jour.
Amer. Med. Assoc, 53 (1909), No. 21, p. 1741). According to

212 Original Communications: Eighth International [vol.

his summary, "the invertebrates are able to convert adenin into
hypoxanthin and guanin into xanthin, showing the presence of
the enzyms, adenase, and guanase, but the metabolism proceeds
no further. Passing upward in the scale of animal life to the
birds and reptiles we find that nitrogen is excreted chiefly in the
form of uric acid. Mammals form uric acid only from the purins
and have the power of destroying some of the uric acid formed.
The enzyms that destroy uric acid seem to be the last formed in
development and are possessed by various mammals in varying
degrees and in the same animal often show an uneven distribution
in the various organs of the body. This uricolytic power is
relatively weak in man." The paper is followed by a discus-
sion.

The output of organic phosphorus in urine was studied by G.
C. Mathison (Bio-Chem. Jour., 4 (1909), No. 5-7, pp. 274-279)
under conditions of work and rest. In young adults, on an
ordinary diet, the organic phosphorus was usually found to be
more than 0.1 gm. per day. Occasionally it fell below this, while
in one case it reached 0.3 gm.

"The percentage of the total P2O6 present in organic combina-
tion varies considerably from day to day. In the cases examined
ii averaged 6 per cent of the total.

"The addition of a large quantity of organic phosphorus in the
form of glycero-phosphoric acid to the diet had no distinct effect
on the output of organic P2OB, while it increased the total PjOs
output. Glycerophosphoric acid was not broken down by gastric
or pancreatic digestion in vitro, so it was probably absorbed
unchanged.

"In the observations made, vigorous exercise was not followed
by increased output in organic P2O6.

"The N:P205 ratio was fairly constant in anyone individual
on a fairly regular diet. It differed greatly in different in-
dividuals, and also in the same individual when the diet was
irregular."

The construe Dion of improved forms of respiration calorimeters
and progress reports of investigations of the Carnegie Institution
of Washington aie contained in the reports of the Nuurition
Laboratory at Boston, which is under the direction of F. G. Bene-

xviii] Congress of Applied Chemistry 213

diet (Carnegie Inst. Washington Year Book, 6 (1907), pp. 130-133;
Carnegie Inst. Washington Year Book, 7 (1908), pp. 158-162,
pi. 1. dgms. 4).

The respiration calorimeters in use at the Nutrition Research
Laboratory of the Carnegie Institution located at Boston, Mass.,
are described in detail by F. G. Benedict and T. M. Carpenter
(Carnegie Inst. Washington Pub. 123, pp. VII-l-102, pis. 5, figs.
25). A general plan is given of the calorimeter laboratory, the
principles involved in the construction of the calorimeter are
considered, descriptions of different parts of the apparatus pre-
sented, and the calculation of results explained. The descrip-
tions are illustrated with diagrams and reproductions of photo-
graphs. An account is given of the routine followed in an experi-
ment with a man as subject. It is hardly possible to give an
adequate account of this work in abstract.

Control tests of a respiration calorimeter, by F. G. Benedict,
J. A. Riche, and L. E. Emmes (Amer. Jour. Physiol., 26 (1910),
No. 1, pp. 1-14), in which alcohol was burned in the respiration
chamber and the amounts of heat eliminated, water vaporized,
carbon dioxid produced, and oxygen consumed were compared
with theoretical values. The results showed that the agree-
ment between measured and theoretical amounts was very
satisfactory and that the apparatus has proved as accurate as
the usual analjrtical methods employed in the laboratory, with
which small amounts of material are studied chemically or
calorimetrically.

The direct determination of oxygen in experiments with the
respiration calorimeter has been compared with calculated val-
ues by F. G. Benedict (Amer. Jour. Physiol., 26 (1910), No. 1,
pp. 15-25), who concludes that the du-ect determination is accu-
rate, "and that experiments on man can be made in which the
direct determination of oxygen is fully substantiated by the indi-
rect determination. Personal experience would indicate that
the errors involved in the indirect determination of oxygen are
such as to preclude its use under conditions that ordinarily ob-
tam in even the most perfect forms of respiration apparatus, and
that accurate determinations of the oxygen consumption of man
are practicable only by means of the direct method."

214 Original Communications: Eighth International [vol.

The influence of muscular and mental work on metabolism
and the efiiciency of the human body as a machine were studied
by F. G. Benedict and T. M. Carpenter (U. S. Dept. Agr., Office
Expt. Stas. Bui. 208, pp. 100, figs. 3) in connection with the nutri-
tion investigations of the Department of Agriculture, the work
being done before the respiration calorimeter was taken from
Middletown, Conn., to Washington. The first of the two papers
included in the bulletin reports data of 19 experiments on the
effects of muscular work on metabolism and the efficiency of the
body as a machine, respiratory products and oxygen consump-
tion being measured in the usual way with the respiration cal-
orimeter, and the bicycle ergometer employed for measurements
of muscular work. The efficiency of the body was found to be
20 per cent; that is, for every calorie of muscular work produced
by the body a total of 5 calories is expended. A series of 44 expe-
riments was made to compare mental work (writing the answers
to examination papers which were regarded as difficult by the
subjects) and mental idleness, perhaps more propeily mental
occupation which involved no special mental effort, namely,
transcribing an amount of very simple material, which gave
them the same amount of arm motion required in writing the
examination papers. No constant differences in the heat out-
put in the different periods were noted, so it appears that the
experiments do not warrant the conclusion that mental work
such as was performed had a positive influence on metabolic
activity measurable by the very delicate methods employed.

Metabolism in man with greatly diminished lung area was
studied by T. M. Carpenter and F. G. Benedict (Amer. Jour.
Physiol., 23 (1909), No. 6, pp. 412-419) at the Carnegie Insti-
tution Nutrition Laboratory, a respiration calorimeter being
used. The only deduction which can be drawn from the exper-
imental data, according to the authors, is' that the reduction
of the area for oxygen absorption and carbonic acid elimination
in the lungs by about one-half did not materially alter the total
metabolism.

During the experiments with the respiration calorimeter made
at Middletown, Conn., by T. M. Carpenter and F. G. Benedict
(Amer. Jour. Physiol., 24 (1909), No. 2', pp. 187-202) several

xvin] Congress of Applied Chemistry 215

cases of illness were observed which, after careful invescigation,
were attributed to poisoning caused by mercury vapor, due to
the use of mercury valves in the ventilating air current.

Metabolism during fever was also studied by the same authors,
with the respiration calorimeter (Amer. Jour. Physiol., 24 (1909),
No. 2, pp 203-233).

"In general the carbon dioxid excretion was apparently greater
during fever than during control periods.

"The oxygen consumption during fever is in practically all
cases noticeably greater than during control. . . .

"While the data show a slight tendency for the respiratory
quotient to increase during fever, the complications attending
the ingestion of food, variations in muscular activity, and errors
in oxygen determination do not warrant any sweeping deduc-
tions from these data."

The recorded data indicate that in general "there was an in-
crease in the water of vaporization during fever over that during
the control period. Since, however, the control experiments
showed marked variations when compared with the fever expe-
riments during periods when there was no appreciable fever, it
is obvious that here again we can not draw any sweeping deduc-
tions regarding this point."

As regards heat elimination, the authors state that " in view
of the necessarily tentative nature of all deductions made from
these experiments, it has not been deemed advisable to attempt
to discuss the influence of fever on the various paths of heat
elimination."

The metabolism of man during the work of typewriting was
studied by T. M. Carpenter and F. G. Benedict (Jour. Biol.
Chem., 6 (1909), No. 3, pp. 271-288), who conclude that "it
seems reasonable to assume that the work of writing some 1,500
to 1,600 words per hour on the typewriter results in an increase
over the resting metabolism of some 10 to 14 gm. of carbon dioxid,
10 to 13 gm. of oxygen, and 20 to 30 calories of heat per hour.
Of these factors of metabolism, it is highly probable that the
truest factor is presented by the total energy exchange as di-
rectly measured, and hence taking into consideration all the data
furnished by these two experiments, we can tentatively say that

216 Original Communications: Eighth International [vol.

the wricing of 1,600 words per hour on the typewriter results in
a heat transformation over and above the resting metabolism of
not far from 25 calories. At present too little is known regard-
ing the energy transformation of various everyday actiAdties to
make any striking comparison, but [by other investigation] . . .
it has been computed that there is an hourly energy expenditure
of about 160 calories over and above the resting maintenance
requirement of a man of 70 kg. walking along a level road at a
rate of 2.7 miles per hour. It is seen, cherefore, that the work
of typewriting calls for very much less transformation of energy
than does that of ordinary walking."

An apparatus for studying respiratory exchange has been de-
scribed by F. G. Benedict (Amer. Jour. Physiol., 24 (1909), No.
3, pp. 345-374, figs. 6), which is similar in principle to that por-'
tion of a respiration calorimeter which has to do with gaseous
exchange and respiratory quotient measurements.

An important digest of data accumulated in experiments with
the respiration calorimeter, extending over 10 years, is included
in a bulletin on the metabolism and energy transformations of
healthy man during rest, by F. G. Benedict and T. M. Carpen-
ter (Carnegie Inst. Washington Pub. 126, pp. VIII -f- 225). Ten-
tative tables are given for computing the metabolism of normal
individuals with varying degrees of muscular activity.

The oable on page 217 gives the carbon dioxid eliminated, the
oxygen absorbed, and the heat produced per hour during various
activities, the data as to standing and very severe muscular exer-
cise being calculated, using as a standard the results obtained
with 55 men awake and sitting up.

"The results presented in this report are to be considered sim-
ply as indicating the normal metabolism of healthy young men
at rest and under several conditions of muscular activity. The
variations from the normal exhibited by the individual can be
seen by an examination of the tables. The attempt is made to
point out the cause of the variations in so far as possible, but
with so complex a process as the energy transformation and cata-
bolism in the body, it is clearly futile to attempt to predict with
great accuracy either the caGabolism or the energy transforma-
tions of a given individual. Approximate values that may prove

XVIIII

Congress of Applied Chemistry

217

Carbon Dioxid Elimination, Oxygen Absorption, and Heat PHODtrcrioN
PER HouB Hour during Various Acttvitibs

Degree of Muscular Activ-
ity

Num-
ber of
subjects

Average

body

weight

Carbon
dioxid
elimi-
nated

Oxygen

ab-
sorbed

Heat
pro-
duced

Man at rest, sleeping

Man at rest, awake, sitting
UD

17
55

Kg.
66.6

64.5

Grama
23

248

Grains
21

213

Calories
71

Man at rest, standing, cal-
culated

114

Man at very severe muscu-
lar exercise, calculated . .

653

of practical use can be obtained by means of some of the factors
outlined in this report. With more accurate and improved cal-
orimeters, there should be in the course of a few years the addi-
tion of many factors, at present entirely unknown."

The influence of the preceding diet on the respiratory quotient
after active digestion has ceased has been studied by F. G. Bene-
dict, L. E. Emmes, and J. A. Riche (Amer. Jour. Physiol., 27
(1911), No. 4, pp. 383-405).

In general, the conclusion was drawn that the respiratory
quotient determined 12 hours after a meal rich in carbohydrates
was higher than when the last meal contained only a small amount
of carbohydrates. The possibility of this high respiratory
quotient being due to the delayed absorption and combustion of
carbohydrates in the alimentary tract is discussed, but the auth-
ors believe that the evidence is rather against the theory.

"Obviously che previous body condition play a very impor-
tanc role. The extent to which the body storage of glycogen
has been drawn upon, the muscular activity of the day previous
to the experiment, possibly the temperature of the surrounding
air, the general diet of the individual for several days before —
in fact, anything which contributes to a disturbance of the stor-
age of glycogen in the body — ^may alter the influence of the in-

218 Original Communications: Eighth International [vol.

gestion of a carbohydrate-rich meal. If the glycogen storage in
the body is at a low poinc, the ingestion of a carbohydrate-rich
meal does not result in an increased respiratory quotient in
accordance with the amount ingested, as a not inconsiderable
proportion of the carbohydrate may be stored immediately as
glycogen. Until this glycogen storage has been replenished the
combustion of carbohydrate in the food may be delayed. On
the other hand, with individuals subsisting without food and
remaining quiet in a respiration chamber, the store of glycogen
may last for some time. From these data we may infer, then,
that muscular activity may play an important role in effecting the
storage of glycogen."

Other questions which have to do with the general subject are
discussed.

The respiratory exchange as affected by body position was
studied at the Carnegie Institution of Washington by L. E.
Emmes and J. A. Riche (Amer. Jour. Physiol., 27 (1911), No. 4,
pp. 406^13). In general, the authors found that the pulse rate
lying down was on an average 63, the carbon dioxid excretion
209 cc, and the oxygen consumption 236 cc. per minute. With
a subject in a sitting position the pulse rate was 71, the carbon
dioxid excretion 218 cc, and the oxygen consumption 254 cc.
per minute.

In their discussion of the data reported the authors point out
that for experimental purposes, when metabolism at a given
condition of body rest is to be determined, it is of value to know,
"as a result of experiments with the respiration apparatus, that
the metabolism of a subject when sitting absolutely quiet in a
chair, without extraneous muscular activity, represents a meta-
bolism 8 per cent greater than that of a subject lying on a couch,
with similar muscular rest. The difference in metabolism is then
due, primarily, to the difference in the internal muscular activity
necessitated by the sustaining of body parts. This is in con-
formity with the well-known fact that the pulse rate of an indi-
vidual when sitting is always noticeably higher than when he is
lying down. From these tests we could infer that if it were pos-
sible to so support the body of the subject in a sitting position
that the pulse rate would be no greater than when the subject

xviii] Congress of Applied Chemistry 219

was lying down, the metabolism would be essentially the same
in both positions."

Using a small apparatus for studying respiratory exchange,
G. Lusk (Amer. Jour. Physiol., 27 (1911), No. 5, pp. 427-437)
investigated the influence of cold baths on the glycogen content
of man.

"Immersion of normal men in cold baths at a temperature of
10° when the intestine is free from carbohydrates induces shiver-
ing, which causes a rapid utilization of body glycogen, as deter-
mined by a fall in the respiratory quotient to the fasting level.
In one very muscular individual this result could not be obtained.

"In one individual in whom the shivering had been severe, a
quotient of 0.67 and another of 0.62 were found during subse-
quent periods of rest, which correspond to those observed during
rest afcer a period of exhaustive exercise (glycogen formation
from protein).

"The greatest increase in heat production which was brought
about by the cold baths was 181 per cent above the normal.
The urine remained free from albumin and from sugar."

A respiration apparatus for the determination of the carbon
dioxid produced by small animals has been constructed at the
Boston Nutrition Laboratory of the Carnegie Institution of
Washington and described by F. G. Benedict and J. Homans
(Amer. Jour. Physiol., 28 (1911), No. 1, pp. 29-48, dgms. 2).

The effects on men at rest of breathing oxygen-rich gas mix-
tures were studied by F. G. Benedict and H. L. Higgins (Amer.
Jour. Physiol., 28 (1911), No. 1, pp. 1-28, fig. 1), with normal
individuals. The air mixtures contained 40, 60, and 90 per cent
oxygen, respectively, and the tests were made in a condition of
complete muscular rest 12 hours after the last meal was taken.
It was found "that there is no apparent difference between the
metabolism as indicated by the gaseous exchange (i.e., the
carbon dioxid output, oxygen consumption, and respiratory
quotient) and the metabolism when breathing ordinary air; that
there is no change in the respiration, either as to character, depth,
or frequency, as compared with the same factors when breathing
ordinary air; [and] that the pulse rate is lower with oxygen-rich
mixtures than when breathing ordinary air; furthermore, that

220 Original Communications: Eighth International [vol.

the higher the percentage of oxygen breathed (up to 90 per cent),
the lower the pulse."

Data regarding the relative ease of digestion of cheese as com-
pared with beef have been reported by C. F. Langworthy and
R. D. Milner (2. Cong. Internat. Hyg. AUment. Bruxelles [Proc],
2 (1910), Sects. 4-7, pp. 249-253). No constant differences in
the heat elimination per hour were noted when comparable
amounts of cheese and meat (beef) were added to a uniform
basal ration. So far as the reported data and resuhs of laoer
experiments not yet reported show, there were no marked
differences with respect to ease of digestion of these two staple
foods.

The success which attended preliminary experiments with
the respiration calorimeter on the possibilities of studying
physiological processes by means of the gaseous exchange and
heac output led to the construction of an instrument of suitable
size for this special work and other related problems. , C. F.
Langworthy and R. D. Milner (U. S. Dept. Agr., Office Expt.
Stas. Circ. 116, pp. 3) have described briefly the new calorimeter
of small size and also the large apparatus used for experiments
with man, as well as a micro-calorimeter for use in the experi-
mental study of very small quantities. The calorimeters as now
installed are equipped with recording and controlling devices of
special construction. The devices of this character used in
connection with the small respiration calorimeter, designed for
the study of vegetable problems, involve much that is new and
original, so that the calorimeter is very largely automatic in
operation. Important modifications have also been introduced
into the construction of the calorimeter itself which make for ease
of operation.

The work which has been done thus far with ripening fruit
has demonstrated that the respiration calorimeter is fully as well
suited to the study of certain fundamental problems of plant life
as to the study of similar problems of animal life.

The apparatus can also be used, it seems safe to conclude, in
studying such problems as the changes which take place when
meat or cheese or other similar products are cured or ripened, and
factors which influence these changes; that is, problems which are

xvm] Congress of Applied Chemistry 221

of commercial interest as well as of agricultural, domescic, and
scientific importance.

The construction of the small respiration calorimeter for use
in the study of problems of vegetable physiology is given by C.
F. Langworthy and R. D. Milner in a later paper (U. S. Dept.
Agr. Yearbook 1911, pp. 491-504).

The respiration of apples and its relation co their keeping
quality were studied by F. W. Morse (New Hampshire Sta. Bui.
135, pp. 85-92, figs. 2). The results of experiments in one
season, calculated on the basis of 1 kg. of fruit for 1 hour, showed
that the average exhalation of carbon dioxid was 18 mg. at
summer temperature, 8.1 mg. at cellar temperature, and 2.7
mg. at cold storage temperature (32 degrees). The apples
obtained in a second series of experiments were 13.2 mg. at
cellar temperature, 21.9 mg. at summer temperature, and 5.2
mg. at cold storage temperature.

The expired breath was studied with leference to the presence
of organic matter, by M. J. Rosenau and H. L. Amoss (Jour.
Med. Research, 25 (1911), No. 1, pp. 35-84, figs. 5). Using the
reaction of anaphylaxis, the authors conclude from experiments
in which the liquid obtained by condensing the moisture from the
expired breath of man was injected into guinea pigs, that the
presence of organic matter in expired breath has been demon-
strated.

"The logical conclusion from our results is that protein sub-
stances under certain circumstances may be volatile. It seems
unlikely that such a complex molecule should possess the power of
passing into the air in a gaseous form. The volatility, however,
now in question may resemble that solubility which deals with
particles in suspension in a physico-chemical state (colloidal
suspension). The protein may simply be carried over in 'solution'
in the water vapor.

"Our experiments are too few to state that albuminous sub-
stances such as egg white, milk, or blood serum in vitro is 'vola-
tile.' However, they are sufficiently suggestive to stimulate
further work along this line."

222 Original Communications: Eighth International [vol.

Foods and Their Relation to Problems ok Hygiene

The need for clean food is much more generally recognized than
was formerly the case. This matter and many others pertaining
to food in its relation to hygiene have been studied during the
period under consideration in the summary.

Studies of the influence of various dietary conditions on
physiological resistance have been reported by N. B. Foster
(Jour. Biol. Chem., 7 (1910), No. 5, pp. 379-419). The results
perhaps pertain more to pharmacology than to dietetics.

The influence of dietary alternations on the types of intestinal
flora was studied experimentally by C. A. Herter and A. I.
Kendall (Jour. Biol. Chem., 7 (1910), No. 3, pp. 203-236, pis.
3), who observed a marked bacterial degeneration following pro-
nounced physiological alternations in the flora of the intestines
as a result of changes in the diet. Experiments were made with
laboratory animals. The authors consider it probable that
analogous conditions would be found to exist in man.

The question has been further discussed by C. A. Herter
(Internat. Beitr. Path. u. Ther. Ernahrungsstor. Stoffw. u.
Verdauungskrank., 1 (1910), No. 3, pp. 275-281) in a paper.

The results of an extended study of fecal bacteria of healthy
men have been reported by W. J. MacNeal, L. L. Latzer, and
J. E. Kerr (Jour. Infect. Diseases, 6 (1909), No. 2, pp. 123-169,
flg. 1). In general, they conclude that "the direct quantitative
determinations of the fecal bacteria furnish evidence of the
extent and nature of the bacterial growth in the intestines. This
seems to be a delicate index of intestinal conditions."

Numerous studies on the effect of lactic acid ferments on
intestinal putrefaction have been reported, including a paper
by Helen Baldwin (Jour. Biol. Chem., 7 (1909), No. 1, pp. 37-48).

Coagulated milk and a large number of other preparations were
used by P. G. Heinemann (Jour. Amer. Med. Assoc, 52 (1909),
No. 5, pp. 372-376) in a study of lactic acid as an agent to induce
intestinal putrefaction.

The results obtained led the author to conclude "that so far as
the therapeutic effect is concerned, there is yet no convincing
evidence that sour milk prepared with commercial cultures is

xviii] Congress of Applied Chemistry 223

preferable to naturally soured milk. Yeasts were present in all
but one of the commercial preparations."

The bacterial condition of protected and unprotected foods
in restaurants, meat markets, grocery stores, bakeshops, and fruit
stores was studied by H. E. Barnard (Ann. Rpt. Bd. Health Ind.,
27 (1908), pp. 517-523, pis. 4), who found that foods kept in glass
cases were in every case practically free from dust and accom-
panying bacteria, while food on exposed tables and racks was
surrounded with air heavily laden with dirt and bacterial life.
It was also found that cleanliness of floors and utensils lessened to
a certain extent the number of bacteria present, and that on the
contrary counters and stands near sidewalks are always sur-
rounded with atmospheric dust and dirt.

The author's studies were concerned chiefly with the relative
number of bacteria found on the culture plates inoculated under
different conditions and the types of bacteria were not thoroughly
differentiated. He believes that pathogenic bacteria were
present.

G. W. Stiles (U. S. Dept. Agr., Bur. Chem. Bui. 136, pp. 53,
figs. 15) studied shellfish contamination from sewage-polluted
waters and from other sources, the observations made in many
localities being supplemented by bacteriological work.

According to the author, "there is undisputed evidence to show
that shellfish become contaminated when placed in sewage-
polluted water, and that Badlluscoli and B. typhosus will
survive for variable lengths of time in the liquor and the body
contents of such shellfish after their removal from infected
water. ... .

"Oyster beds should be protected from every possible source
of contamination, and they should be located in water proven
to be pure by repeated examinations. . . .

"The practice of floating oysters in water of questionable
purity should be absolutely prohibited because of the probability
of sewage contamination. . . .

"Like other perishable food products, oysters may become
unfit for use if stored or kept under insanitary conditions. This
spoilage, however, may take place wholly from the length of time
out of water. . . .

224 Original Communications: Eighth International [vol.

"The liquor in the shell surrounding the oysters contains more
bacteria than does an equal volume of meat from the same oyster.
This liquor, together with any sand in the gills of the oyster, can
be removed and the meat chilled at the same time by the use of
pure ice and water. This washing process can be done efficiently
within 3 to 10 minutes, depending upon the method employed.
Oysters should not be allowed to soak in fresh water, as they
increase in volume, change in appearance and flavor, .and decom-
pose more rapidly than those not soaked.

" [As shown by cooking tescs], steaming contaminated oysters
and clams in the shell, or coolcing them after shucking for 15
minutes at boiling temperature, practically destroys all organ-
isms of a questionable character, but since in practice shellfish are
never cooked for this length of time, cooking can not be depended
upon to remove this danger. . . .

"The investigations show that vast areas of valuable shellfish
grounds in this country are now reasonably free from sewage
pollution, but this territory will gradually diminish in size if
sewage is not properly cared for in the future. Comparatively
speaking, only a small acreage is now subject to serious pollution."

The absorption of aluminum from aluminized food was studied
by M. Steel (Amer. Jour. Physiol., 28 (1911), No. 2, pp. 94-102).
When alum was administered in aluminum-free foods to dogs or
when they were given biscuits baked with alum baking powder,
"aluminum in comparatively large amounts promptly passed
into the blood.

"Absorbed aluminum circulated freely, but as it did not show
any pronounced tendency to accumulate in the blood, its full
effects must have registered outside of the circulation."

When aluminum chlorid was administered intravenously, from
5.55 to 11.11 per cent of the aluminum passed from the blood
into the feces during the 3 days immediately following the injec-
tion. "AVhether the aluminum passed directly through the walls
of the intestine or was excreted by the Uver, or whether both
channels (or others) were followed, has not yet been ascertained."

The use of metallic containers for edible fats and oils was
studied by J. A. Emery (U. S. Dept. Agr., Bur. Anim. Indus.
Rpt. 1909, pp. 265-282), of the Bureau of Animal Industry, with

xviii] Congress of Applied Chemistry 225

vessels and sheets of tin plate, galvanized iron, copper, tin, lead,
zinc, aluminum, and iron, for the purpose of determining the
action of fats and oils upon metals, with particular reference to
the utility of these metals as containers.

The work shows that where an increase in the acid content of
the fat or oil was noted there was an increase in the solvent action
of the oil for metals, particularly where other favorable condi-
tions, such as heat, moisture, and exposure to the atmosphere,
were present. With cotton-seed oil, however, an exception was
noted, as this oil, when prepared with a corn oil of lesser or ap-
proximately the same acidity, showed little or no effect upon
metals.

"It [was] demonstrated that zinc, copper, and lead are some-
what readily acted upon, while aluminum, iron, and tin, in the
order in which they are named, have offered evidences of higher
resisting power and are the metals which would more satisfac-
torily meet the requirements of both manufacturer and con-
sumer."

Much attention has been given in the United States to the
discussion and study of pellagra, on account of its supposed rela-
tion to Indian corn. The agricultural aspects of the pellagra
problem in the United States were studied by C. L. Alsberg (N.Y.
Med. Jour, and Phila. Med. Jour., 90 (1909), No. 2, pp. 50-54).
Of important discussions may be mentioned the report of C. H.
Lavinder (Pub. Health and Mar. Hosp. Serv. U. S., Pub. Health
Rpts., 24 (1909), No. 37, pp. 1315-1321), and the paper on the
etiology of pellagra by the same author (N. Y. Med. Jour, and
Phil. Med. Jour., 90 (1909), No. 2, pp. 54-58), who has also re-
ported considerable data regarding pellagra and its possible rela-
tion to maize according to some recent views (Pub. Health and
Mar. Hosp. Serv. U. S. Pub. Health Rpts., 26 (1911), No. 8, pp.
199-208). Particular interest attaches to his discussion of Rau-
bitschek's photodynamic theory that the disease is ascribable
to the joint action of a substance present in corn meal fat and
sunlight.

W. H. Buhlig (111. Bd. Health Mo. Bui., 5 (1909), No. 7, pp.
417-435, figs. 2) and J. F. Siler and H. J. Nichols (111. Bd. Health
Mo. Bui., 5 (1909), No. 7, pp. 437-478, figs. 8) have made exten-

226 Original Communications: Eighth International [vol.

sive studies of the possible relation between corn in the diet and
the occurrence of pellagra. Final conclusions were not drawn.
In connection with this work some data were reported on expe-
riments in cookery as well as regarding institution dietetics.

Cost of Living and Other Statistical Data

The collection of statistical data has continued as an important
part of the general activity in nutrition. Much of the work in the
United States has been done under government or state auspices.

A select list of references on the cost of living and prices, by
H. H. B. Meyer (Washington: Library of Congress, 1910, pp.
V+107), has been published by the Library of Congress.

A large amount of statistical data on wages and prices of com-
modities has been reported in various Senate documents (Wash-
ington: U. S. Senate Select Committee, ,1910, vols. 1, pp. 658;
2, pp. Ill-f 659-875 — Hearings held before the Select Committee
of the Senate relative to wages and prices of commodities). Topi-
cal digest of evidence submitted in hearings held before the Select
Committee of the Senate relative to wages and prices of com-
modities (Washington: U. S. Senate Select Committee, 1910,
pp. XCV).

In connection with the work of the U. S. Census many studies
have been reported on the production and value of food products;
for instance, those on rice cleaning and polishing, by H. McK.
Fulgham (Bur. of the Census [U. S.] Bui. 61, pp. 49-58, dgm. 1),
and beet sugar, by Z. C. Elkin (Bur. of the Census [U. S.] Bui.
61, pp. .'>9-69); slaughtering and meat packing (Bur. of the Cen-
sus [U. S.] Bui. 83, pp. 7-41; and starch, by R. H. Merriam (Bur.
of the Census [U. S.] Bui. 64).

The Bureau of Labor has published a summary of retail prices
of food from 1890 to 1906 and discussed the data with reference
to the cost of living in the United States (Bur. of Labor [U. S.]
Bui. 71, pp. 175-328).

A large amount of data regarding the prices of foodstuffs in
different New Jersey cities and towns has been summarized in an
article on the cost of living in New Jersey (Ann. Rpt. Bur. Statis.
Labor and Indus. N. J., 30 (1907), pp. 141-157).

xviii] Congress of Applied Chemistry 227

Data regarding the prices of meat in the United States have
been summarized in the Yearbook of the Department of Agri-
culture, by the Secretary, James Wilson (U. S. Dept. Agr. Rpts.
1909, pp. 15-31; Rpt. 91, pp. 10-24; Yearbook 1909, pp. 15-31).

Much statistical data regarding Hawaiian honey are included
in a paper on Hawaiian bee keeping, by E. F. Phillips -U. S.
Dept. Agr., Bur. Ent. Bui. 75, pt. 5, pp. 43-58, pis. 6).

Information regarding the cost of living of wage-earners and
other similar material, including a paper on the preparation of
a rational diet at a reasonable cost, are incorporated in the report
of the Committee on Social Betterment, by G. M. Kober (Re-
ports of the President's Homes Commission. Washington, D.C.,
1908, [pt. 5], pp. 281, pis. 4; Reprint, pp. 281, pis. 4; see also
Alimentation and foods (U. S. Senate, 60th Cong., 2. Session
Doc. 644, pp. 121-157).

R. C. Chapin (New York, 1909, pp. 372, dgms. 16) has reported
an extended investigation of the standard of living among work-
ingmen's families in New York City, which presents and dis-
cusses a large amount of statistical data.

The cost of living in American towns has been exhaustively
studied by the British Government (London: Govt., 1911, pp.
XCn-F533, map 1; U. S. Senate, 62 Cong., 1. Sess., Doc. 22,
pp. XCII-l-533), the results being presented in a report by H. L.
Smith which was reprinted by the U. S. Senate, and summarized
in publications of the U. S. Senate (U. S. Senate, 62 Cong., 1.
Sess., Doc. 38, pp. 74) and of the Department of Commerce and
Labor (U. S. Dept. Com. and Labor, Bur. Labor Bui. 93, pp.
500-570).

Included in the study were 28 American towns on or east of
the Mississippi, with an aggregate population in 1910 of 15,500,-
000, in round numbers.

The range of price levels for rents was found to vary greatly,
being the highest in New York City. The prices of the prin-
cipal foodstuffs, such as bread, flour, meat, potatoes, and sugar,
did not show great range in the different towns, as was evident
from the fact that when each article is considered in its relative
importance the lowest level is 91 and the highest 109, with New
York midway counting as 100. "If the towns are grouped geo-

228 Original Communications: Eighth International [vol.

graphically the New England and southern groups show the
highest food price levels, the Midle West towns the lowest, the
position of the New England towns in regard both to wages and
rents being here reversed."

As regards retail prices of foods, the conclusion is that the
ratio between the United States and England and Wales is 138
to 100.

"One peculiarity shown by the budgets is the comparatively
small consumption of baker's bread in the average American
working-class family, the consumption being 8| lbs. weekly per
family as against 22 lbs. in the United Kingdom, the place of
bread being taken in the United States to some extent by rolls,
cakes, biscuits, eoc, on which the expenditure is about three
times as great as that shown in the average British budget. On
the other hand, the consumption of meat is much larger in the
United States, and the consumption of vegetables is also larger.
The budgets indicate in general that the dietary of American
working-class families is more liberal and more varied than that
of corresponding families in the United Kingdom."

In addition to general discussions the reporc contains the de-
tails of the family budgets and other statistical data collected.

CONCLCSION

In the foregoing summary of work in human nutrition which
has been carried on in the United States since the Seventh Inter-
national Congress of Apphed Chemistry, the attempt has been
made to give some idea of che general condition of nutrition in-
vestigations and to cite examples of investigations along the
proper lines of work into which the subject naturally divides itself.

That the list of investigations is by no means complete is rec-
ognized but it is believed that enough has been brought together
to show that progress has been continuous and to make it clear
that important contributions have been made not only to the
fund of available data of interest to the students of nutrition
and to practical workers, but also to mechods of investigations
as well as to the more important matter of fundamental theories
of nutrition.

AN IMPROVED FORM OF RESPIRATION CALORI-
METER FOR THE STUDY OF PROBLEMS OF
VEGETABLE PHYSIOLOGY

By C. F. Langworthy and R. D. Milner

Nvirition Investigations, Office 0/ Experiment Stations, Dept. of

Agriculture, Washington, D. C.

Theoretical considerations regarding ripening fruit led to the
attempt to study such questions of plant life by methods which
have given good results in investigations of topics pertaining to
the nutrition and energy expenditure of man. It was found that
ripening fruit (bananas) could be studied in this way, since when
they were kept during the active ripening period in the chamber
of the large respiration calorimeter described in a publication of
the Department of Agriculture,^ carbon dioxid and water vapor
were given off and oxygen was absorbed and heat liberated, all
in measurable quantities. In other words, conditions were
present which could be studied with great exactness with the aid
of this apparatus.

It was furthermore apparent from the results obtained in this
preliminary work that many other problems of plant 'life could
be studied by such methods and that results of both practical
and scientific value could be secured, since a knowledge of the
factors determined is of great importance in the consideration of
questions pertaining to vegetable metabolism and to the handling
and storage of fruit in the home and under commercial conditions.

The chamber of the respiration calorimeter used for experi-
ments with man is of such a size that it will accommodate seven
or more large bunches of bananas; that is, its capacity is too great
to make it useful for experiments with vegetable products, except
those which can be obtained in uniform condition and in fairly
large quantities. Furthermore, the entrance to the respiration
calorimeter and all its internal arrangements are designed with

'U. S. Dept. Agr. Yearbook 1910, p. 307.

229

230 Original Communications: Eighth International [vol.

respect to experiments with man and are not particularly well
suited to experiments with fruits or similar products. It was
obvious, therefore, that a smaller respiration calorimeter with
special equipment suited to experiments with plant products
would be a great convenience and in view of the fact that it would
be useful in the study of problems of interest to the Department
of Agriculture, such an instrument was constructed. In plan
and principle it corresponds to the large respiration calorimeter
used for experiments with man although some improvements in
grouping of accessory apparatus have been introduced and some
new automatic regulating devices which greatly lessen the labor
of conducting the experiments have been added, which make for
greater accuracy as well as ease of operation.

A respiration calorimeter is thus designated because such an
instrument combines in the same device a respiration apparatus
for the determination of gaseous exchange of the subject in the
respiration chamber during a given period, and a calorimeter for
measuring heat liberated in the respiration chamber. Though
there are conditions under which it is convenient to use the
apparatus either as a calorimeter only, or as a respiration appara-
tus only, the two operations are usually carried on simultaneously.
The two functions of the apparatus, however, are perhaps best
considered separately.

The Apparatus as Used for Studying Respiratory Exchange

The chamber of the apparatus is 18 by 18 by 36 inches inside
measurements, the walls being copper attached to a wooden
framework, and double for purposes which are explained later.
The top of the chamber has a cover that may be lifted off so that
material may be put inside. The edges of the cover are formed
so as to fit into grooves in the upper edges of the sides, and by
means of a special wax (mixture of beeswax and Venice turpen-
tine) pressed into the grooves the cover is sealed tight. In the
upper part of each of two opposite sides is a window 6 inches by
8 inches sealed into its frame, which forms part of the copper wall.
These afford opportunity to observe the fruit during ripening, or
to remove a sample if desired. In the third wall is an "outlet,"
likewise sealed air-tight, through which pass two f inch pipes for

xviii] Congress of Applied Chemistry 231

the passage of air into and out of the chamber. On the inside the
pipe carrying the ingoing air opens near the top of the chamber,
while that carrying outgoing air is extended to the bottom, so
that air passsing from one opening to the other must traverse the
chamber.

The chamber is of sufficient size to accommodate a large bunch
of bananas suspended from a framework supported on brackets
in the corners near the top ; and other brackets have been attached
at different levels so that trays, shelves, or other supports may be
used. Thus it is possible to place under investigation, at different
times, not only a bunch of bananas, but also larger or smaller
quantities of apples, or potatoes, or other materials. This
material may be packed in the calorimeter in a manner approxi-
mating commercial conditions, or, if desired, in some other
manner.

On the outside the pipe for outgoing air is connected with a
rotary compressor, operated by a small electric motor which
withdraws the air from the chamber and forces it any desired
rate through a purifying system of special gas washing bottles
arranged in series. The first two bottles of the series contain
sulphuric acid, which removes from the air all water vapor brought
out by it from the chamber. The next two bottles contain
granular soda lime (a mixture of caustic soda and quick lime)
which removes from the air all the carbon dioxid carried out of
the chamber. Following these is another bottle of acid which
catches moisture imparted to the dry air by the moist soda lime.
The final bottle in the series contains granular sodium carbonate,
which catches any sulphuric acid vapor or spray that might be
carried from the preceding bottle. The air then returns to the
chamber through the pipe for ingoing air. Just before entering
the chamber oxygen may be admitted to the ingoing air to replace
that used by the material in the chamber.

A small copper pipe, from the side of the chamber, is connected
outside with a rubber bag or a spirometer or similar device, the
purpose of which is to indicate the volume at any given time, and
to maintain atmospheric pressure within the chamber.

The different bottles of the purifying system are weighed on
a large sensitive balance to an accuracy of 0.05 of a gram. The

232 Original Communications: Eighth International [vol.

increase in weight of the first two, which contain sulphuric
acid, shows how much water was removed from the chamber
in the air current during a given experimental period, and that of
the two soda lime bottles and of the acid bottle following them
shows the amount of carbon dioxid brought out. Under ordinary
conditions, in an experiment with fruit, these would represent
the amounts actually produced during the period, though when-
ever necessary samples of the air may be analyzed to determine
the amounts residual in the air of the chamber, and allowance
may be made for any difference in these amounts at the begin-
ning and the end of the period.

With a ventilating system such as described above, as rapidly
as any gaseous substance is removed other gas must be introduced
to maintain atmospheric pressure in the chamber. Ordinarily,
oxygen is admitted, since oxygen is utilized by the ripening fruit
or other material from the air of the chamber. In case the
fruit were ripened in some inert gas, such as carbon dioxid or
nitrogen, this gas would be admitted instead of oxygen. The
gas admitted to the chamber is drawn from a supply under
pressure in a steel cylinder that is suspended on a sensitive
balance, and the amount admitted is determined very accurately
by weighing. In the case of oxygen, for example, from the
change in the weight of the cyhnder and in the amount residual
in the air at the beginning and the end of the period, the amount
used by the fruit during the period may be ascertained.

Experimental periods may be of any duration. Since the
ripening process continues for several days in the case of bananas,
it has been sufficient in these experiments to weigh the different
bottles in the purifying system once a day.

The Apparatus as a Calorimeter

The heat generated by the ripening fruit or other material in
the respiration chamber is carried out by a current of cold water
flowing through a coil of copper pipe. From the weight of the
water flowing through the coil during a given period, and the
mean temperature difference between the incoming and outgoing
water during the period, the quantity of heat carried out is
determined. This quantity and that carried out as latent heat

Photoeraph No. 1.— This is a view of the exterior of the respiration calorimeter for the
study of vegetable problems. The protective covering is in place and the window in
one side is shown.

Photograph No. 2. — ^This is a view of the respiration calorimeter showing how portions
of the protective covering may be removed. The cold-water pipes for cooling the air
in the space between the protective covering and the walls of the respiration chamber
are also shown. The cover of the respiration chamber is raised. On the wall panel
are shown switches for various electric currents, the preheater for warming, and the
bridge for determining the temperature of the water entering the heat absorbing sys-
tem, and other devices.

Photograph No. 3. — In the lower right-hand corner of this photograph is shown the
automatic temperature recording device with its pen which draws the Une represent-
ing temperature differences. Next to this on the left is shown the bridge pertaining
to this device, by means of which it is possible to vary the range of the records and
to test the accuracy of the recording device. Immediately above the recorder is
shown a device for the automatic control of the temperature of the water which
enters the heat absorbing system in the respiration chamber. At the left on the
same shelf is a device which automatically controls the heating of the air adjacent to
the top, sides, and bottom of the exterior wall of the respiration chamber and of the
ingoing air in the circulating air system. The bridge for this device is shown on the
shelf immediately below it.

xvni] Congress of Applied Chemistry 233

of water vapor in the out going air represent, with small correc-
tions for changes in temperature of the chamber and of material
inside, the amount of heat generated by the material in the
chamber.

The quantity of water that flows through the coil in the cham-
ber is ascertained by weighing. Water is kept flowing through
the coil within the chamber at as constant a rate as possible.
For this purpose the water is drawn from a constant level tank
on a shelf above the calorimeter. Since distilled water is used
in the circulating system, after it has passed from the calorimeter,
it is collected in a tank from which, by means of a small gear
pump, it is raised again to the constant level tank. This small
pump is driven by the motor which operates the rotary com-
pressor for the circulating air. The purpose of using distilled
water in this cooling system is to avoid difficulties due to a
presence of air in the water, which tends to collect in bubbles
in the pipes and thus form temporary obstructions, causing an
irregular flow of water from the heat absorbing system.

The temperature of the air in the calorimeter may be regulated
by controlling either the temperature of the water that enters
the calorimeter, or its rate of flow, or both. The most convenient
practice is to maintain a constant rate of flow and to regulate
only the temperature. To this end the water leaving the pressure
tank is cooled by passing it through a coil immersed in cold water
or brine and then is raised to the desired temperature by means of
an electric heating device introduced in the water circuit just
before it enters the calorimeter. The ingoing water is kept at
such temperature that the heat will be taken up just as fast as
it is liberated, so that the temperature of the air in the chamber
is kept constant.

This heating device consists of a pre-heater and of a final
heater. The pre-heater, which is operated manually, has a total
capacity of 5f° C, with a rate of 500 cubic centimeters of water
per minute, and heat may be added as needed in small increments.
To secure uniformity of temperature, the final heater is separated
from the pre-heater by a mixing bottle. The final heater, which is
automatic, has a much smaller range than the pre-heater, but it
can be adjusted within very narrow limits. The sensitive

234 Original Communications: Eighth International [vol.

portion of the final heater consists of a very delicate electrical
resistance thermometer, of a type designed by the National
Bureau of Standards, which comprises one arm of a Wheatstone
bridge, the slide wire of which is designed to cover by tenths of a
degree a range of temperature from 0° to 35°C., shown on the dial
of the bridge. This thermometer is placed in the water pipe
immediately above a small heating coil of electric resistance
wire, so that the water flows from the coil directly over the
thermometer. If the temperature of the water flowing over the
thermometer differs from the desired temperature, at which
the pointer of the bridge is set, the needle of the galvanometer
with which the bridge is connected is deflected accordingly; and
the automatic controller, of which the galvanometer forms a
part, alters the position of a sliding contact on a variable resist-
ance which is in series with the heater just below the thermometer.
This results in a change in the amount of electric current through
the heater, its heating effect is varied accordingly, and the tem-
perature of the water flowing past the thermometer is regulated
until it reaches the desired constant temperature.

The difference between the temperature of the water entering
the calorimeter and that leaving is determined by means of a
pair of electric resistance thermometers placed in the water line,
one just as it enters and the other just as it leaves the chamber,
and connected with a Leeds and Northrup temperature recorder.
This temperatiu-e recorder consists of a self-balancing Wheat-
stone bridge, two arms of which are formed by the resistance
thermometers. The amount of change necessary in the balancing
point of contact on the slide wire to balance the bridge at any
given time is indicated by a pen that is drawn back and forth on a
record sheet that moves forward at a rate of 2| inches per hour.
Since the balance is determined by the mechanism every five
seconds, a virtually continuous record of the temperature differ-
ences is drawn by the pen. The scale on the paper is 10 inches
wide and represents a total range of 2°. The scale is ruled with
100 lines, each representing 0.02°, but the distance between the
Unes is so wide that 0.01° is very easily read. The bridge part
of the apparatus is so constructed that the slide wire may be made

xviu] Congress of Applied Chemistry 235

to represent a temperature difference of from 0 to 2°, 1 to 3°,
2 to 4°, or 3 to 5°.

In order that the quantity of heat generated within the calori-
meter chamber may be accurately determined, it is necessary to
prevent either gain or loss of heat through the walls of the
chamber. To this end the respiration chamber has double metal
walls and the outer wall is kept at exactly the same temperature
as the inner wall, in which case there will be no transference of
heat between them. In order to accomplish this it is necessary
to provide means for determining any difference of temperature
between the two walls and for heating or cooling one wall until
it has the same temperature as the other. In this calorimeter
the outer metal wall is surrounded by a covering of heat insulating
material, between which and the metal wall is an air space about
1 inch across. In this space and surrounding the outer wall of
the chamber are a coil of copper pipe to carry cold water for
cooling it, and a coil of resistance wire to carry an electric current
for heating it. If the outer wall of the calorimeter is too warm it
may be cooled by passing cold water through the copper pipe,
or if it is too cold it may be heated by passing an electric current
through the resistance wire; but in practice it is found most
convenient to allow water to flow continually through the coil
of pipe and to vary only the heating. This is done auto-
matically.

On the inner and outer copper walls of the calorimeter cham-
ber are electric resistance thermometers which comprise the
two arms of a Wheatstone bridge, which have exactly the same
resistance when the walls are at the same temperature. As
the temperature of one wall varies from that of the other the
resistance of the arms of the bridge varies and this causes a
corresponding change in a mechanism which controls a variable
resistance in series with the heating system surrounding the
outer wall, and thus regulates the heating of the outer wall.
By this means the temperature of the exterior wall is maintained
automatically in balance with that of the interior wall. The con-
trolling mechanism regulates the temperatme of the top, sides,
and bottom of the chamber independently. The temperature of
the air entering the calorimeter is likewise maintained by it

236 Original Communications: Eighth International [vo

exactly the same as that leaving the calorimeter, so that no he;
will be carried in or out in the moving air current.

Possible Uses of the New Respiration Calorimeter

The control experiments and the experimental studi^ ^
ripening fruit (bananas) already undertaken have demonstrate
the great accuracy of this respiration calorimeter as an instrumei
of precision and have given interesting results regarding gaseoi
exchange and heat production which will appear in Departmei
of Agriculture publications.

Although the new calorimeter is arranged with special referenc
to experiments with fruits and other vegetable products, it is s
constructed that the respiration chamber can be removed ar
another substituted for it of the same size but with differei
interior arrangements, or of smaller size, should this be desirabl
In other words, it would be possible, with little additional labo
since no change in the recording and controlling devices and oth(
accessory apparatus would be involved, to adapt the apparati
to the study of additional problems, such, for instance, as tl
incubation of eggs and the changes which take place in curir
and storing meat products and cheese, or by making suitab
provision for the collection of excretory products and for tl
comfort of the subjects, it would be possible to adapt the calor
meter to experiments with laboratory animals, should the woi
of the Department make this necessary.

SUR LE ROLE ANTISEPTIQUE DU SEL MARIN ET

DU SUCRE

Par M. L. Lindet
Paris, France

U est facile de concevoir comment certains corps qui sont
pour nous des poisons, comme les composes de I'arsenic et du
mercure, peuvent arrSter le d^veloppement des microbes; mais
Taction du sel marin et du sucre, dont nous faisons im usage
journalier, me semble ne pas avoir €t6 suffisamment envisag^e.

Elle s'explique cependant par la facility avec laquelle les mi-
crobes se plasmolysent; ils cedent k une solution concentr6e de sel
ou de sucre une partie de leurs 616ments constitutifs, s'affaiblis-
sent, et ne pr6sentent plus la m^me capacity de reproduction.

J'ai voulu rechercher dans quelle mesure la composition des
microbes est capable de se modifier sous I'influence de solutions
sucr6es ou salines, de concentration variable, et j'ai choisi celui
des microbes qu'il est le plus facile de se procurer en masse, la
levure de distillerie; celle-ci provenait de la distillerie Springer
k Maisons-Alfort (Seine). Dans le but de mesurer la sensi-
bility du ph6nom6ne, je n'ai laiss6 la levure en contact de la
solution que pendant 24 heures, et j'ai dos6 I'azote, I'acide
phosphorique et la potasse dans les liquides filtr^s. J'ai rap-
ports les chiffres obtenus k la quantity de matiSres que la levure
contenait primitivement:

Azote

% des 416ments contenus
daas la levure:

Acide phos-
phorique

Potasse
(KO.)

Ttooin Eau pure . .

1.89
1.99
2.19
2.65
1.89

(t
ti

11.13

0.57
1.32
1.60
1.77
1.78
5.33

11.38

73.3

Solution de sel k 2%

75.4

Solution de sel k 4%

77.8

Solution de sel k 8%

82.1

T6moin Eau pure .

73.3

Solution de sucre k 20%

92.6

Solution de sucre k 40%

93.8

Solution de sucre k 80%

96.8

237

238 Original Communications: Eighth International [vol.

Evidemment les quantit^s d'azote et d'acide phosphorique
dont la cellule s'est appauvrie ne sont pas tr^s considerables,
surtout en presence de la solution de sel; mais il faut songer
qu'elles repr^sentent les mati^res les plus solubles de la cellule,
celles que la cellule mettra en jeu d^s les premiers moments de
son Evolution. La solubility des composes potassiques au con-
traire leur confSre un coefBcient de diffusion considerable.

L'etude au microscope des levures ainsi soumises k Taction des
solutions salines ou sucr^es r^veie avec nettete leur amaigrisse-
ment.

En presence de ces faits, il 6tait int&essant de rechercher
comment se reproduisent, sur bouillon de touraillons, g^latind
et Sucre des globules de levure qui out sejourne 48 heures au
contact des memes solutions. J'ai applique, pour la numeration
des levures la technique que j'ai expose dans un precedent
travail (Comptes-rendus de I'Academie des Sciences, 1910, T.
150, p. 802), et j'ai rapporte le nombre des colonies comptees au
mmg. de levure.

Colonies par mmg. de levure :

Temoia 4.514.000

Solution de sel a, 5% 4.370.000

Solution de sel k 10% 1.733.000

Solution de sel k 20% 600.000

Solution de sucre k 20% . .• 1 .525.000

II convient en outre de faire remarquer que les colonies de ces
differentes levures ont apparu sur la gelatine avec un retard
d'autant plus grand qu'elles avaient sejourne au contact de
solutions plus concentrees. Une fois apparues, elles n'ont pas
augmente sensiblement en nombre du jour au lendemain; mais
celles qui ont ete formees au debut ont grossi reguli^rement, k
fur et k mesure qu'elles retrouvaient dans le bouillon gelatine
les elements qu'elle avaient perdus.

J'ai commence des experiences analogues avec le ferment
lactique et avec les champignons; mais ces experiences sont
plus difficiles k reahser, et je demande credit pour quelque
temps.

SALMON CANNING INDUSTRY OF NORTH AMERICA

H. M. LooMis
Chief of the Food and Drug Inspection Laboratory, Bureau of
Chemistry, U S. Dept. Agriculture, Arcade Annex
Building, Seattle, Wash.

The salmon of the North Pacific Ocean has now become one of
the most important marine food products on this continent and
its popularity is fast increasing in Europe and other countries.
The catching and packing of salmon in the Northwest has devel-
oped into such a large industry that it ranks second only to the
lumber business. As the growth has been so rapid some apprehen-
sion has been felt that the fish might be gradually exterminated
but the Federal government and the governments of the various
states and of Canada are striving to overcome any such danger
by regulating the industry and by establishing hatcheries at
various favorable localities.

There are five principal varieties of salmon packed along the
Pacific Coast, each one of which is known by several names,
depending on the locality where it is caught. The fish with the
reddest flesh and most oil are held in the highest esteem by
consumers, and in the following list they are given in the com-
monly accepted order of quality.

1. Red Salmon, Sockeye, or Blueback.

2. Chinook, King or Spring Salmon.

3. Medium Red Salmon, Cohoe or Silverside.

4. Humpback or Pink Salmon.

5. Chum or Dog Salmon.

The 1911 pack of salmon amounted to 290,000,000 cans or six
million cases. Of this entire output Alaska produces nearly one
half, Puget Sound district about one quarter, British Columbia
one sixth, Columbia one twelfth and the balance is caught in
various rivers and bays along the coast of California, Oregon and
Washington.

239

240 Original Communications: Eighth International [vol.

For 14 years the United Kingdom has taken an average of
over 900,000 cases a year while Australia, the East Indies and
South America make continually increasing demands on the
product.

The habitat and history of the Sockeye and other salmon is
unknown from the time they first reach the sea as young fish
until they return to spawn and die — a period of about four years.
These fish are not caught by hook and line but in seines aod
traps of various forms. The traps are located along the shores
of the mainland or islands at points which the large schools of
fish pass on their journey from the ocean to the rivers, and each
trap consists of a row of piling, running out from the shore a
distance of several hundred feet. To this, nets are hung ver-
tically to a considerable depth below the water, which depth is
regulated by law. These serve to divert the course of the fish
into the trap proper, which are rectangular enclosures, formed of
nets supported vertically by piles and into which the fish are
directed by V-shaped openings. To empty the trap a sort of
apron net is raised horizontally until the fish are near the sur-
face and then a bail net is used to transfer the fish to a scow or
steamer. As many as 50,000 fish are sometimes taken in a trap
at once.

The Sockeye Salmon is comparatively small, weighing from
5 to 10 lbs., while other varieties of salmon are larger — the largest
variety, the Chinook, averaging 30 lbs.

Until very recently the older type of soldered can was used for
packing salmon but the solderless, so-called sanitary can, is
rapidly growing in favor and in 1911 about 1,700,000 cases of
fish were packed in the latter form of cans.

To manufacturing chemists it may be of interest to know that
in this industry alone 840,000 lbs. of hydrochloric acid, 180,000
lbs. of caustic soda, 6,000,000 lbs. of solder, 137,000,000 sq. ft. of
tin plate, and 375,000 gallons of lacquer are used.

Of the 325,000,000 lbs. of salmon caught last year about 225,-
000,000 were canned and the rest were cured in fight brine,
frozen, salted or smoked.

I wish to make acknowledgment to "The Pacific Fisherman"
for most of the statistics given above.

xviii] Congress of Applied Chemistry 241

At the cannery the fish are unloaded and carried by conveyors
to:

1. The "iron chink" — a machine which removes the heads,
tails, fins, and entrails.

The further steps in the canning process may be briefly enu-
merated:

2. Cleaning.

3. Washing.

4. Slicing by a machine which cuts the fish transversely into
pieces the right size to fit the cans.

5. Adding of salt to cans.

6. Packing of fish in cans. This is usually done by white
women in the United States and by Indians in Alaska.

7. Covers put on, crimped and soldered by machine.

8. Cans cooled and vent hole soldered by hand.

9. Cans tested for leaks by immersing in a hot water bath.

10. Placed in steam retort for 1-2 hour.

11. While still hot, covers are punctured, allowing most of the
air and some of the liquor to escape. The sound made by strik-
ing the can also serves to detect leaks.

12. Cans resealed.

13. Cans heated in steam retorts about one hour at 240° to
cook and sterilize contents.

14. Cans are scoured with caustic soda solution and washed.

15. Cans tested for leaks from the sound emitted in tapping
the cover.

16. Cans lacquered and labeled.

After the process of canning and before shipment every can
is usually tested several times for leaks.

In the United States canneries the labor is almost entirely
done by Chinese or Japanese men — the transferring of the fish to
the cans employing white women. In Alaska, owing to the
scarcity of white labor, most of the carmeries employ Orientals
and Indians exclusively.

It might be well to mention that the low form of soldered cans,
or "flats," are filled by hand, while the tall cans, or "tails," are

242 Original Communications: Eighth International [vol.

filled by machinery and the latter contain a product which is
inferior in appearance and price to the former.

In packing the so-called sanitary cans, the number of steps in
the process is considerably less and, while the cans are more expen-
sive in the first place, the saving in solder, labor, and other items
amounts to about 25c a case. With this form of can there are
no vents and the filled cans are run on conveyors through a steam
chest, (to heat the contents and expel air), covers are crimped
immediately and the cans placed in the retort for final cooking
and sterilization.

The methods in use for preparing canned salmon in the United
States are generally adapted to the production of a fresh, clean
and high grade product.

Since the passage of the National Food and Drugs Act and
promulgation of Food Inspection Decision No. 105, regarding the
labeling of canned salmon, misbranding is rarely resorted to and
the cans are generally labeled to show the variety of salmon con-
tained in them. The public is further safeguarded in the label-
ing of salmon by the provisions of the Alaska Fisheries Law of
June 26, 1906, requiring that in the labeling of canned salmon no
false or misleading statement or designation shall be made. The
enforcement of this law is in the hands of the Bureau of Fisheries.

A proper study of the composition of canned salmon requires the
analysis of many samples of known history, for the different
varieties of salmon vary in composition in different parts of the
body and at different seasons, the earlier runs of fish being much
fatter and finer than the later.

Only a few analyses are reported herewith, but they represent
the beginning of a more extended investigation of the composition
and quality of fresh and canned salmon at the Seattle Laboratory
of the Bureau of Chemistry. The fresh salmon were analyzed
within 24 hours after being taken from the traps and were kept
on ice as much as possible during that time.

The following notes on the analytical methods seem necessary:

Ammoniacal nitrogen: — For this determination two similar
methods were employed. The first method is substantially the
alcoholic distillation method of Richardson and Scherubel,

xvni] Congress of Applied Chemistry 243

(J. A. C. S., Vol. 30, page 1515), with certain modifications
proposed by W. B. Smith. 450 cc. of 95% alcohol by volume were
used instead of 60% alcohol, with 25 grams of material and 5
grams of freshly igaited maguesium oxide and the fish was added
directly to the distilliag flask without previous extraction. 750
cc. Kjeldahl flasks were used and three 150 cc. portions were
distilled into N-10 acid, making up the volume in the distilling
flask to 450 cc. with 95% alcohol between each portion. The
excess of acid is titrated with N-10 caustic soda and cochineal
indicator.

The second method was to place 25 grams of fish in a 500 cc.
Florence flask, add 5 grams magnesium oxide and 100 cc. of 95%
alcohol by volume and distill in a current of boiling 95% alcohol
vapor, using an apparatus like that shown on page 37 of Gatter-
mann's "Practical Methods of Organic Chemistry." 400 cc.
of distillate were collected in N-10 acid and the excess of acid
titrated as above.

In both methods blank determinations were made with the
reagents and the necessary correction applied.

244 Original Communications: Eighth International [vol.

CANNED SALMON
1911 pack

WATER

ETHYL
ETHER

EX-
TRACT

PRO-
TEIN

(Nx
6.25)

TOTAL
ASH

NaCl.

AMMONIACAL
NITROGEN

Richard-
son
method

Alcohol

vapor

method

No. 1.

Puget Soimd
Sockeye Sal-
mon

62.44

15.17

20.25

2.50

0.79

0.0403

0.0348

No. 2.

Puget Sound
Sockeye Sal-
mon

61.84

13.74

21.77

2.73

1.10

0.0437

0.0410

No. 3.

Alaska Me-
dium Red
Salmon

69.97

7.81

20.40

2.58

1.09

0.04965

No. 4.

Alaska Chum
Salmon

73.48

2.88

21.33

2.57

0.83

0.0563

0.0557

No. 5.

Alaska Pink
or Hump-
back Salmon

74.12

4.75

19.75

1.98

0.50

.0404

No. 6.

Alaska Red
Salmon

70.88

5.26

21.79

2.35

0.64

.0455

Each sample was average of two or more cans.

AH samples, except No. 2, were old form 1 lb. tall cans.

No. 2 was i lb. flat cans.

xviu]

Congress of Applied Chemistry

245

ANALYSES OF FRESH SALMON, EDIBLE PORTIONS

WATER

ETHYL
ETHER

EX-
TRACT

PRO-
TEIN

(Nx
6.25)

TOTAL
ASH

NaCl

AMMONIACAL
NITROGEN

Richard-
son
method

Alcohol

vapor

method

Puget Sound
Sockeye Sal-
mon, caught
May 7, 1912.

67.48

8.86

22.24

1.36

0.0121

0.0205

Puget Sound
Steelhead
or Sal-
mon Trout,
caught May
7, 1912

67.89

9.39

21.80

1.35

0.0135

0.0218

From a comparison of the fresh and canned Puget Sound sal-
mon there is evidently considerable reduction in water content
during the canning process. As all samples of canned salmon
were in good condition and gave no indication of deterioration
as far as the senses could detect it, the results on " ammoniacal
nitrogen" are also of interest, being two or more times greater
in the case of the canned product than in the fresh fish.

PROPOSED METHOD FOR THE ESTIMATION OF TIN
IN CANNED FOODS

By H. L. Lourib
Bureau of Chem., V. S. Appraiser's Stores, New York, N. Y.

Immediately before and subsequent to the issue of Food
Inspection Decision 126, which limits the amount of tin in canned
foods to less than 300 milligrams per kilogram, it was necessary
in the course of routine work in the New York Laboratory, for
a large number of analyses for tin to be made.

The first method used was the Munson Combustion Method
as given in Bulletin 107, U. S. Dept. Agriculture. This method
had to be discarded because of its length, and its doubtful ac-
curacy in the case of canned foods containing salt, as then there
was a loss of volatile tin salts during the combustion. It thus
became necessary to find a method that would fulfill two
conditions :

I. Accubacy: II. Rapidity:

The first method tried was practically that described in the
report to the Local Government Board of England, by Drs.
Buchanan and Schryver relative to the presence of tin in certain
caimed foods, published at London, 1908. In this method the
organic material is destroyed as in a nitrogen determination by
means of potassium sulphate and concentrated sulphuric acid.
While this proved accurate enough, it was discarded because of
its tediousness and the constant breaking of flasks. I attempted
a modification of this method by using potassium permanganate
in conjunction with the sulphuric acid. This was not entirely
successful because of the large amounts of permanganate necessary
and the constant attention it required. Finally a method was
tried using nitric and sulphuric acids to destroy the organic mat-
ter. This proved successful from the start, not only being
rapid, but also yielding practically 100% recovery, in the case of
known amounts of tin. The method was developed not only for

247

248 Original Communications: Eighth International [vol.

canned materials such as fish, vegetables, fruits, etc., but also
for foods high in sugars, such as maple syrup, molasses, jam, etc.,
Below are directions for each class:

Directions fob Material such as Fish

Place 25 to 100 grams of the well mixed and finely ground-
sample (the quantity employed depending upon the amount of
fat or oil present) into a kjeldal flask (800-1000 c.c), and add 25
to 50 c.c. of concentrated sulphuric acid, the amount depending
upon the weight of the charge. Place the flask on a hot plate or
on wire gauze over free flame; add about 30 c.c. of concentrated
nitric acid, raise the temperature to boil and heat till white fumes
are generated, then without cooling add 10 c.c. of nitric acid and
continue heating as before. Repeat the nitric acid addition until
the solution remains clear (usually straw color), after boihng
off the nitric acid fumes. The digestion can be easily accom-
plished in about one hour with three or four additions of nitric
acid. Let the solution cool, and dilute to about 400 c.c. with
water. Neutralize with concentrated ammonia, transfer the
solution to a beaker, rinse out flask with a little concentrated
ammonia, add to main solution, make slightly acid with sulphur
ric, and saturate with HzS gas. Let the precipitate settle on
steam bath, filter, wash with a Httle hot water saturated with
HaS, and then dissolve the precipitate in hot yellow ammonium
sulphide. Reprecipitate with acetic acid or hydrochloric acid,
filter on ashless paper, ignite, moisten with nitric acid, ignite and
weigh as stannic oxide. SnOa.

Directions for Material such as Strtjp

Weigh 50 to 100 grams in kjeldal flask (800-1000 c.c.) and add
about 100 c.c. of water and 150 c.c. concentrated nitric acid.
Boil until all the fumes are driven off, then add a few c.c. more of
nitric acid, and boil to see if there is any further action. Repeat
addition of nitric acid and boiling until there is no further action.
Then add concentrated sulphuric acid, a few c.c. at a time,
heating until all the nitrous acid fumes are driven off. When
20 to 25 c.c. of sulphuric acid have thus been added, boil until

xviii] Congress of Applied Chemistry 249

sulphuric acid fumes are driven off. Now add concentrated
nitric acid, five c.c. at a time, until the solution is clear. Then
proceed as in case of canned goods given above.

Note: Fifty c.c. of concentrated ammonia will nearly neutralize
25 c.c. concentrated sulphuric acid. Make usual tests for com-
plete precipitation in the filtrate from the first tin sulphide
precipitate. In the case of canned vegetables, as high as 100
grams may be taken without using more than 50 c.c. of sulphuric
acid. With fish it is best to take as many c.c. of sulphuric acid
grams as grams of fish. The rapidity of the digestion depends
on the temperature maintained — the higher the temperature, the
faster the material is oxidized.

ON THE PREPARATION OF "NATTO"

S. MURAMATSU

College of Agriculture, Morioka, Japan

There are several kinds of natto prepared in Japan, but here
I mean common natto which is a kind of vegetable cheese made by
fermenting boiled soya beans wrapped in rice straw and set in a
warm cellar for one or two days. Thus the product becomes
white and mucilageous by the development of bacteria. Natto
is consumed as an accessory after having been mixed with table
salt and several stimulants, amongst others the powdered mus-
tard is preferred. It is chiefly consumed in Tokyo and the
north-eastern districts of Japan and for the production of it Aizu
is the noted place. It is chiefly consumed in Tokyo in the
summer time, but in the north-east during the winter time, as
these are rather poor in vegetables at that season.

There exist several studies on natto so far as to its constituents
and the micro-organisms forming it, but no exact investigation is
known of about its preparation. So, its manufacturers suffer
under many difficulties of preparing natto of good quality; for
this reason, I was obliged to make a study of the method of pre-
paring it and several other points. Besides, I think it is very
useful to prepare natto of good quality and increase its consump-
tion by the people, as it is a very good and economical food stuff,
being cheap and containing much protein, especially in our coun-
try where rice is the principal food.

I. Soya Beans

Soya beans are the principal raw material of natto. There are
numerous varieties of soya beans cultivated in Japan, which,
for instance, we can distinguish by their color as yellowish
white, green, black, spotted, etc. I prepared natto with these
different kinds and could not find a more suitable kind than the
small yellowish white bean.

261

252 Original Communications: Eighth International [vol.

The beans which serve for the preparation of natto are first
sorted and all that are broken or imperfectly developed are
picked out; besides, it is better to sift them through sieves with
proper meshes to separate too small or too large ones. They are
then washed and allowed to steep in clean water for several hours,
after that they are boiled in a large iron kettle with sufficient
water for ca. 5 hours. Thus the beans become moderately soft
and their color darker.

Their constitution was as follows:

In 100 pts. air-dry beans: —
Moisture 7 . 14

Dry matter 92.86

In 100 pts. dry matter : —
Crude protein 50 . 156

Crude fat 22.453

Crude fibre 6.420

N-free extract 11 . 871 f Soluble in water 4 . 329

\ Insoluble in water 7 . 542
Ash 3.600

Total-N 8.025

Albuminoid-N 7 . 953 f Soluble in water — trace

I Insoluble in water 7 . 953

Non-albuminoid-N

0.072

II. Rice Stkaw

Rice straw is used for the wrapper of the boiled soya beans.
Fresh straw is preferable to old, as its smell is better than that of
the latter. The straw is cleaned by taking the muddy leaf away
from the imder part of the stem and washed with clean water;
afterwards it is well tied at its two ends, leaving several inches
apart and bundled after filling the bag with the beans. As to
the reason of using straw for the preparation of natto, it was
considered that the straw supplies the proper bacteria to the
beans but I do not think this the sole reason, for we can prepare it
another way, as, for instance, by setting it in a sterihzed Petri-
dish or in a basket. When it is made in a basket, which after
filling it with beans is put in a warm cellar covered with a straw

xvra] Congress of Applied Chemistry 253

mat, it is called hasket-natto. From this and other facts it is
reasonable to consider the principal objects of using straw for
the preparation of natto to be : —

1. To supply the good aroma of straw to natto.

2. To take away ammonia from natto.

3. To offer good ventilation of air to the loosely packed beans.

The bacteria which produce natto from soya beans are always
present on the surface of the beans and their spores being very
strong against high temperature, they are not easily killed by
boiling, as we can see from the following experiment: The grains
which were boiled for several hours are taken in sterilized Petri-
dishes after each hour and placed in the incubator at 42°C. By
this means, I found that the beans which were boiled for 8 hours
become natto rich in mucilage and with good aroma.

The fact that the hsisket-natto, which does not come in touch
with straw, does not sell as well as common natto, for, when we
prepare it in the straw bundle its flavor is always superior to one
which is made in Petri-dish, as it contains an aroma somewhat
like that of straw. So I think that the straw which is used as a
bag for the beans gives its good aroma to natto.

When the bacteria grow on the beans they produce so much
ammonia that we can perceive it by its peculiar smell. As the
straw absorbs ammonia, the smell of it is more feeble when we
use straw bundle than in the case of glass dish. We can under-
stand this fact when we see that the straw which has been used
as a bag always contains much more ammonia than the same
fresh material, and natto, made in the dish is richer in it than that
from bundle.

Amovmt of ammonia
In the fresh straw 0 . 035%

In the straw used as wrapper 0 . 065%

In natto made in a glass dish 0 . 235%

In natto made in straw bimdle 0 . 188%

For these reasons, natto prepared in straw bundle must have
better flavor than any other, by taking its flavor from straw
and giving off the disagreeable smell of ammonia to straw.

254 Original Communications: Eighth International [vol.

The bacteria producing natto want much oxygen for their
proper growth, as it is an obligate aerobe. So, when we prepare
it by heaping up many bundles the interior ones become inferior
in quaUty and also the interior beans of a large bundle become not
so viscous as the outer parts. For this reason, it is recommend-
able to use small bundles for the preparation of superior natto.

III. Cellar

The cellar for the preparation of natto is made with bricks or
with pillars surrounding them with thick layers of straw and
plastering the walls with mud; the entrance is furnished with a
thick door preventing the entering of air. Along the inside of
the wall a long shelf two feet wide is set up at the height of ca.
two feet and one or two large hearths are made on the floor for
the purpose of warming the room.

IV. The Pbepaeation of Natto

For the preparation of natto the soya beans are sorted at first
and all beans that are broken or imperfectly developed are picked
out. After washing with clean water, they are soaked for several
hours and boiled in an iron kettle until they become moderately
soft (ca. 5 hours). The boiled beans are put into the straw bun-
dle while they are still hot and the bundles are placed, standing
obliquely, on the shelf in the cellar, which is previously warmed
by charcoal to about 40°C. The cellar is then shut up carefully,
avoiding the ventilation of air; thus, the beans become natto after
one or two days and are ready for consumption.

V. The Microbes op Natto

As to the micro-organisms of natto several authors have made
investigations. Dr. Yabe isolated three species of micrococci
which formed yellow, orange, and white colonies respectively, and
a bacillus which is not motile, liquefying gelatine and producing a
greenish fluorescence. He attributed the production of the char-
acteristic aroma of natto to the development of the micrococcus
which produces yellow colonies; but no explanation was given
about the formation of the viscous substance.

xvni] Congress of Applied Chemistry 255

Dr. Sawamura isolated various kinds of bacilli and micrococci
from natto and regarded the following two bacilli as the chief
microbes for the production of natto.

Bacillus No. 1. is a motile and facultative aerobe. Natto
produced by this bacillus had a good taste and aroma, but its
viscosity was not so great as that produced by the other. The
author gave the name of Bacillus natto to this bacillus, considering
it as the chief microbe in the fermentation.

Bacillus No. 2 is a rarely motile and facultative aerobe. Natto
produced by this bacillus showed a stronger viscosity but a less
nice taste and aroma than that produced by the B. natto; he recog-
nised it as a variety of Bac. mes. vulgatus. Thus, he concluded
that for the formation of good natto both bacilli must be present.

Mr. Monzen isolated several kinds of bacteria, among them
one bacillus to which Dr. Omori gave the name of Bacillus visco-
sus natto and which he said, is the principal microbe that produces
strong viscosity. The two kinds of bacilli which he named
Bacillus odorans natto I and Bacillus odorans natto II, produce
good aroma in natto; and another one which he named pseudo-
monas odorans natto, produces also good aroma. The latter
three did not produce good natto, unless the material is inoculated
also with B. viscosus natto. Thus the author concluded that there
are necessary for the preparation of natto at least two kinds of
bacteria, one producing the peculiar aroma and the other strong
viscosity.

Mr. Muto isolated several bacteria and concludes that only
one bacillus belonging to B. subtiUs group is necessary for the
production of natto.

I isolated also several bacteria from natto, prepared in Tokyo,
Aizu, and Morioka, and found that these all contain the same
micro-organisms, amongst which the following three bacilli are
the principal ones; several other bacilli are not suitable for the
preparation of natto, as they produce bad color or smell and make
the natto unfit for eating. Two micrococci were foimd, one of
which was analogous to Mic. flavus, and the other producing a
translucent colony on agar plate-culture; but, both the micrococci
having no relation to the preparation of natto, I gave up their
further investigation.

256 Original Communications: Eighth International [vol.

Bacillus No. 1

This bacillus develops most energetically at high temperature
(40 — 50° C.) and produces the best quality of natto, providing
much mucilage and good aroma.
Form:

The cells grown in bouillon at 40° C. are 1 fi thick and 5-8 fi long.

It moves energetically, providing long cilia around its body.
Spore :

An oval spore is formed principally in one end of the cell, which
is 0.8 /tt thick and 1,.6 /* long; the formation of spore requires 4
hours at 42° C. and germination of it begins equatorial after 2|
hours at the same temperature.
Oxygen:

Obligate aerobe.
Coloring:

It is colored readily with aniline coloring matters and also
after Gram's method.
Bouillon culture:

Bouillon remains almost clear after its development, and a
strong folded film, colored shghtly grayish brown, is formed after
10 hours at 38° C.

Sugar bouillon becomes slightly turbid changing its reaction
to acidic at the beginning, which turns alkaline gradually; gas is
not formed.
Peptone-water culture:

It produces a grayish white film on its surface and the liquid
becomes slightly turbid.
Gelatine plate-culture :

Small white colonies are formed which liquefy it quickly.
Gelatine stab-culture:

It develops vigorously at the surface and liquefies gelatine in
the shape of a funnel; the liquefied part remains transparent and
a film is formed.
Agar plate-culture:

White and mealy-looking colony, that has a rough wristle at
its centre but delicate at its edge, spreading very rapidly at 40° C.
Agar slope-culture:

xvin] Congress of Applied Chemistry 257

Colony develops along the line and spreads rapidly all over the
surface with mealy appearance; the condensed water remains
transparent with a film on its surface, but no sediment.
Potato culture:

Elevated colony is formed in the beginning, which spreads soon
over the whole surface of the medium; the colour of the colony is
yellowish brown and it is folded with mealy appearance, the
medium becoming brown.
Milk culture :

It is coagulated at first and is dissolved again.
Hjs:

Is formed.
Indol reaction:

Is not obtained from old bouillon culture.
Reducing property:

It reduces methylene blue in bouillon but does not develop in
the glucose nitrate medium.
Ammonia:

Is formed in the culture of bouillon and Soya beans.
Enzym :

Diastase and proteolytic enzym of tryptic nature are recognised.
Behaviour to temperature:

It develops very vigorously at 50° C, but not at 60° C. It is
killed at 60° C. after two hours, and after one hour at 80° C.

The resistance of the spores against heat is very strong, for it
wants one hour to be killed in Koch's steam-steriliser.
Behaviour to several compounds:
Table salt:

In bouillon containing 15% NaCl it develops slowly, but not in
20%solution.
Alcohol :

It develops in bouillon containing 4%, but not in 5% alcohol.

The spore is not killed readily with alcohol, as it is yet alive
after ten days and more, when put either in 50% or absolute
alcohol at 20° C.
HCl:

It develops in bouillon containing 0.025% HCl, but not in 0.05%
The spore which is put in_3%^HCl is^^alive after one day, but not

258 Original Communications: Eighth International [vol.

after two days. In 4% HCl it is alive after one bour, but not after
two hours.
Acetic acid:

It develops in bouillon with same concentration as hydrochloric
acid.

The spore is not killed by glacial acetic acid after 10 days and
more.
NaOH:

It develops in bouillon containing 0.2% NaOH, but not in
0.3%.

The spore is killed when it is put in 35% solution after one day.
Phenol:

It develops in bouillon containing 0.1% Phenol, but not in
0.2%.

The spore is not killed after ten days when it is put in 5%
solution.
Corrosive sublimate:

It develops in bouillon containing 0.0025% HgCl2, but not in
0.005%.

The spore is killed after 50 minutes when it is put in 0.1%
solution, but it was alive after 40 minutes in the same solu-
tion.

This bacillus may be the same as those which Dr. Sawamura
represented as Bacillus No. 2 and Bacillus viscosus Omori, and
also that which Mr. Muto thought was the only bacterium which
produces natto, though there are several differences in its behav-
iour investigated by these authors.

Bacillus No. 2

This bacillus develops most energetically at high temperature
and produces natto of the best quality, forming much mucilage
and rather higher aroma than Bacillus No. 1.
Form:

The cells grown in bouillon at 40° C. are 0.8-1 /* thick and 4-10 /i
long.
Motility:

It moves vigorously providing long cilia aroimd its body.

xviii] Congress of Applied Chemistry 259

Spore:

An oval spore is formed in one end of the cell, and it is 0.8 /i
thick and 2 /* long.

The spore wants 4 hours at 42° C. for its formation and it germi-
nates equatorial after 2^ hours at the same temperature.
Oxygen:

Obligate aerobe.
Colouring:

The cell is coloured easily by aniline colouring matters and also
after Gram's method.
Bouillon culture:

Bouillon remains almost clear after its development and a
strongly folded film of slightly grayish brown is formed after
twelve hours at 38° C. Sugar bouillon becomes slightly turbid,
changing acidic at the beginning which turns alkaline gradually;
gas is not formed.
Peptone-water culture:

It produces a grayish white film on its surface and the liquid
becomes turbid slightly.
Gelatine stab-culture :

It develops on the surface quickly and liquefies gelatine in the
shape of a funnel; the liquefied part remains transparent and a
film is formed.
Agar plate-culture:

There is formed a white and mealy-looking colony with rough
wristle at its centre but delicate at its edgp, spreads very quickly
at 40° C.
Agar streak-culture :

Colony develops along the line and spreads rapidly all over the
surface with mealy appearance. The condensed water remains
transparent with a folded film on its surface but no sediment.
Potato culture:

Elevated colony is formed in the beginning, which soon spreads
over the whole surface of the medium; the colony is folded and
has brownish yellow colour and mealy appearance ; the medium
becomes brownish gray.
Milk culture :

It is coagulated at the beginning and is dissolved again.

260 Original Communications: Eighth International [vol.

Is not formed.
Indol reaction:

Is not obtained from old bouillon culture.
Reducing property:

It reduces methylene blue in bouillon and produces ammonia
by reduction of nitric acid in the glucose nitrate medium.
Ammonia:

It is formed in the cvdture of bouillon and soya beans.
Enzym:

Diastase and proteolytic enzym of the tryptic nature are
recognised. *

Concerning the behaviour against heat and several compounds
as formerly mentioned, there is not much difference with Bacillus
No. 1.

This bacillus may be the same as that which Dr. Sawamura
named Bacillus natto, thoiigh there are several differences in its
behaviour investigated by us. As this bacillus does not produce
any mucilage at low temperature (say, 35° C.) he thought it,
perhaps, to be one which produces aroma peculiar to natto; but,
as I mentioned already, this bacillus produces much mucilage at
higher temperature and makes good natto with high aroma.

Bacillus No. S

This bacillus develops most energetically at 40° C, and when it
is developed on boiled soya beans at this temperature, it produces
good natto with strong viscosity and good aroma; but its mucilage
is somewhat less than Bacillus No. 1 and Bacillus No. 2.
Form:

The cells grown in bouillon at 40° C. are 1.2 /i* thick and 6-10
IJ' long.
Motility:

It moves providing long ciha around its body.
Spore:

An oval spore is formed in one end of the cell, which is 1 A* thick
and 1.5 A* long.

The spore is formed after 4 hours at 42° C. and germinates
equatorial after 2| hours at the same temperature.

xviii] Congress of Applied Chemistry 261

Oxygen:

Obligate aerobe.
Colouring:

It is coloured readily with aniline colouring matters and also
after Gram's method.
Bouillon culture:

Bouillon becomes slightly turbid and a brittle film of sUghtly
grayish brown colour is formed after ten hours at 38° C. and
produces a small amount of sediment. The film is broken easily
by shaking and sinks to the bottom. Sugar bouillon changes to
slightly acidic at the beginning and turns slightly alkaline after-
wards.
Peptone-water culture :

It becomes sUghtly tmbid and forms a yellowish white film on
its surface. Gas is not formed.
Gelatine plate-culture :

Small white colonies are formed which Uquefy it quickly.
Gelatine stab-culture:

It develops on the surface at the beginning and Uquefies gela-
tine in the shape of a funnel, afterwards thoroughly.
Agar plate-culture:

White colony with rough wristle at its centre but delicate at its
edge, spreads very rapidly at 40° C.
Agar slope-culture:

The colony develops along the Une and spreads rapidly in the
shape of a feather; the condensed water is transparent with a film
on its surface, but no sediment.
Potato culture:

Yellowish gray colony is formed, somewhat elevated in the
beginning; it spreads soon over the whole surface of the medium.
The colony has strong viscosity and it is folded shallower than
Bacillus No. 1. and Bacillus No. 2., the medium becoming gray.
Milk culture:

It is coagulated and dissolved again.
HsS:

Is produced.
Indol reaction:

Is not obtained from old bouillon culture.

262 Original Communications: Eighth International [vol.

Reducing property:

It reduces methylene blue in bouillon, and ammonia is formed
by the reduction of nitric acid in the glucose nitrate medium.
Ammonia:

Is formed in the culture of bouillon and soya beans.
Enzym:

Diastase and proteolytic enzym of tryptic nature are recognized.

The behavior to heat and several compoimds is almost the
same as with Bacillus No. 1, although there are some differences.

This may be the same bacillus as Bacillus grossus, but as there
is no detailed description of it, I cannot make a precise comparison.

VI. The Application of Ctjltuked Bacteria for the
Preparation of Natto

As mentioned already, when we prepare natto in a glass dish at
ca.3.0°C. inoculated with Bacillus No. 1 it has some viscosity,
while others have not, but the aroma was inferior to that made in
straw bundles, for it does not touch with straw. At 45°C. all
bacilh produce natto of fine quaUty providing strong viscosity
and good aropia; the aroma produced by Bacillus No. 1 was the
best, while Bacillus No. 2 produces a rather strong smell of
ammonia, and Bacillus No. S being the worst; moreover, I pre-
pared natto according to the common way differing only on the
point of inoculating these bacilh separately and also mixing them
with one another. The result was that natto which was pro-
duced by the inoculation of Bacillus No. 1 was the best, as it has
much mucilage and fine aroma, while Bacillus No. 2 produced
an inferior and Bacillus No. 8 the worst quality. Natto produced
by the inoculation of mixed baccilli was not so good as that pro-
duced by each bacillus; so, there is no necessity that two or more
bacilli present for the formation of good natto. By the inoculation
of cultured bacteria we can entirely avoid failures and can
prepare good natto by selecting the bacteria. Otherwise, it is
sufficient to put it in the cellar for only one day, after which the
natto will be ready for consumption. So, I recommend to use
the pure culture of proper bacteria according to the following
way:

xvin]

Congress of Applied Chemistry

263

The bacteria developed on the slope culture medium of agar
are mixed with juice produced by the boihng of beans. This is
poured over the surface of boiled beans while they are still in the
kettle, the further process being the same as usual. There is no
necessity of mixing several bacilli.

VII. Natto as a Food Accessort

As natto is prepared from soya beans which are rich in protein
and carbohydrates, it contains much protein and carbohydrates;
the nutritive value of it is greater than that of boiled soya beans,
for it is rich in soluble matters produced by the micro-organisms.

The composition of natto differs exceedingly with age, but its
mean composition is as follows : (Compare with the composition of
boiled soya beans.)

Moisture
Dry matter

Crude protein
Crude fat
Crude fibre
N-free extract

Ash

Total-N

Albuminoid-N

In 100 pts. of fresh natto.

63.480

46.520

In 100 pts. of dry matter:

46.088

20.216

6.140

3.348

Soluble in water 2 . 495
Insoluble in water 0 . 853

5.010 "

7.374

5 . 458 r Soluble in water 1 . 141
\ Insoluble in water 4.317

Non-albuminoid-N 1.916

The micro-organisms which grow on the soya beans secrete
trypsin and diastase; so, when we take it together with several
foods rich in protein or starch, they may be digested more rapidly
than when they are taken alone.

I express many thanks to Dr. Sato, Director of our College,
who helped me in determining the quality of natto that I prepared,
and also to Mr. N. Nitta and Mr. Y. Tanaka who assisted me in
these investigations.

CONTRIBUTION TO THE CHEMISTRY OF THE
RIPENING OF "SHIOKARA"

By Y. Okuda
College of Agriculture, Imperial University, Tokyo

Although the isolation and identification of some nitrogenous
compounds in "Shiokara" has been undertaken about two
years ago by Prof. U. Suzuki, Yoneyama and Otake in this
laboratory, no chemical investigation abo\it the ripening process
of this interesting food material has yet been reported. So I
have tried to contribute something on this line. I have observed
that the autolysis and the action of microbes are two indispen-
sable factors for the preparation of "Shiokara."' Some trials
have also been made to isolate the enzymes which play an im-
portant r61e in this process, and finally, I have carried out some
quantitative determinations to see the chemical changes at
different stages of ripening.

I. Autolysis and the Action of Microbes

1). To see whether the autolysis is going on during the
ripening process of "Shiokara," very fresh organs' of a bonito
fish were minced with a meat-chopping machine, and rubbed
with some quartz sand in a mortar. 40g of the paste thus pre-
pared was divided into two equal parts, and put in the flasks
A and B. After adding 100 c.c. of water to each flask, A was
boiled for a few minutes to destroy the enzymatic action. Both
flasks were then shaken with enough toluol and a little chloro-
form, and kept for 4 days at ordinary temperature. No bac-
terial growth was observed during that time. The flask B was

' "Shiokara" made from the organs of bonito was used.
• The Btomach, the intestines, and the pyloric coecum.
10 265

266 Original Communications: Eighth International [vol.

now boiled, and the contents of both flasks were then filtered
and analysed with the following results : —

A (boiled)

B (not boiled)

Total soluble-N

1.697%
0.184 "
1.513 "

1.895%

Soluble Alb.-N

0.141 "

Non-alb.-N

1.754 "

2). The fresh "Shiokara" two days after preparation was
chopped and crushed in a mortar, 200g of the paste were di-
vided into two equal parts and put in two flasks of 1 litre capacity,
and stoppered with cotton plugs. After adding 500 c.c. water,
one flask was boiled. To each flask was now added enough
toluol and chloroform and after keeping for 4 days at room
temperature, the contents of both flasks were filtered and analysed.

A (Boiled)

B (not boiled)

Total aoluble-N

1.848%
0.056 "
1.792 "
0.604 "

2.052%

Soluble Alb.-N

0.038 "

Non-alb.— N

2.014 "

Amino-N (after fonnol method)

1.023 "

We see from the above two experiments that autolysis is going
on in the fresh organs of bonito fish, and also in the freshly pre-
pared "Shiokara."

3). The microbes, predominating in "Shiokara" seem to be
quite different at different stages of its ripening. In three prepa-
rations, made in April and two months old, we found immense
niunbers of yeasts, bacilli and cocci, but only few moulds, while
in a sample prepared early in October and about one and a half
months old, were found numerous yeasts, the other microbes
being relatively very few.

The isolation and identification of these microbes will be
reported afterwards.

4). 120g of the "Shiokara", which was two months old,
were well crushed and equally diA^ded into three Erlenmeyer

XVIIl]

Congress of Applied Chemistry

267

flasks containing each 100 c.c. of saturated natrium chloride
solution and treated in the following way : —

A. Control: — Not boiled, no antiseptics added.

B. Not boiled, toluol and chloroform added to prevent the
bacterial growth, but not the enzymatic action.

C. Boiled and antiseptics added to prevent both bacterial
and enzymatic action.

After keeping for ten days at 34 — 38°, they were boiled and
filtered, and the filtrates were analysed with the following
results: —

A
(Control)

(Not boiled,

but antiseptic

added)

C
(Boiled and an-
tiseptic added)

Total soluble-N

2.404%
0.049 "
2.355"

2.305%
0.090 "
2.215 "

2.305%

Soluble alb.-N

0.184"

Non-alb.-N

2.121 "

The above experiment shows that both autolysis and the action
of microbes are going on very slowly in the old preparations
compared to fresh ones. The investigation of Wehmer' on
salted herring has shown that the action of microbes upon
proteins is somewhat retarded in 5 per cent, common salt solu-
tion, but it does not entirely stop even in 30% solution. As the
concentration of the salt in "Shiokara" usually is 15%, there is
no doubt that the microbes can still play an important role on
the ripening process, especially at the early stage of its prepa-
ration.

II. Enzymes m 'Shiokara"

Trypsin, diastase and lipase were identified in the fresh organs
of a bonito fish and also in the fresh preparations of "Shiokara."
In the old preparation, however, their action seems to be much
retarded. This observation agrees well with the experiments
mentioned above.

'Wehmer: Abhandlungen des deutschen Seefischerei — Vereins, III, 1898.

268 Original Communications: Eighth International [vol.

1). Fresh organs. The stomach, intestines and pyloric coecum
of a fresh bonito were freed from their contents and rubbed with
some quartz sand in a mortar, and filtered through the cloth
filter. The faintly acid extract thus obtained has shown its
peptonifying power upon milk and fibrin, either in the faintly
acid reaction or after addition of 0 . 2% sodium carbonate. But
no action was observed in presence of 0.2% hydrochloric acid
in the medium, thus the absence of a pepsin is most profitable.

The existence of diastase was shown by the saccharification of
starch paste and glycogen in the neutral reaction.

For the detection of lipase, the minced and groimd organ was
extracted with a mixture of 90 parts of pure glycerine and 10
parts of 1% sodium carbonate, 10 c.c. of the mixture being used
for 1 g of the sample. The liquid was filtered through a piece of
cloth and exactly neutralized. By the addition of some milk or
olive oil to this extract, the increase of acidity due to the formation
of fatty acids by the action of Hpase upon neutral fats was ob-
served. Of course some toluol and chloroform being added to
prevent the bacterial growth.

2). "Shiokara" at different stages of ripening. The following
observation was made with the samples collected at different
stages of ripening: —

(a). " Shiokara," two days old.

Trypsin. Present, active.

Diastase. Do.

Lipase. Present, but the action was very weak,

(b). "Shiokara." 40-50 days old.

Trypsin. Present, but very weak.

Diastase. No reaction.

Pepsin. Do.

(c). "Shiokara," 50-60 days old.

Trypsin. Very weak.

Lipase. Do.

Diastase. No reaction.

3). Isolation of enzymes. For this purpose, about 200g of
the fresh sample, 3 days old, were finely minced and ground
with some quartz sand in a mortar and macerated with a little

xviii] Congress of Applied Chemistry 269

distilled water. The liquid was strained through linen cloth,
and after dialysing for about two hours to get rid of the greater
part of the common salt, it was poured in a mixture of absolute
alcohol and ether, the grayish white voluminous precipitate
thus produced was then collected on a filter, washed with abso-
lute alcohol and ether, and dried over sulphuric acid. The
crude enzyme preparation obtained in this way, when dissolved
in a little water has shown strong diastatic and tryptic action
while that of lipase was very weak. When the solution of this
crude enzymes was added to a solution of various amino-acid,
no liberation of ammonia was observed, showing the absence
of amidase.

The proteolytic enzyme which acts in weak alkaline as well as
in neutral or in faintly acid reaction, but not in a 0 . 2% hydro-
chloric acid solution, was also found by Blanchard* in several
fishes and by Roaf* in two crustaceae.

' Blanchard, Jahresberioht flir Their-Chemie, 13, 1883— Orig. Compt. rend.
96, 1241.

"Roaf, Jahresber. f. Tier Chem. 36, 1906 — Orig. Biochem. Journal, 1. 390-
87.

270 Original Communications: Eighth International [vol.

III. Chemical Changes During the Ripening Process

1). The sample^ used for this detefmination was prepared on
the 17th of June, 1911, and after 3, 6, 12, 25, and 40 days re-
spectively a portion was taken for analysis. Thus the following
results were obtained: —

Date of analysis

Days after
prepara-
tion

In 100 parts of fresh samples

Water

Dry matter

Total-N

Alb.-N

Ether-extract

Soluble matter

Non-alb.-N

Ammonium-N

Organic base-N

Othei-N

Total acid (as lactic)

NaCl (calculated from total
chlorine)

64.96
35.05
1.98
0.35
1.83
27.25
1.63
0.15
0.84
0.74
1.43

17.31

64.78
35.22
2.04
0.35
1.81
28.45
1.69
0.15
0.72
0.80
1.42

64.58
35.42
2.05
0.28
1.81
29.24
1.83
0.25
0.69
1.09
0.97

17.34

64.25
35.75

0.26
1.71
31.10
1.
0.18
0.63
1.17
0.96

63.99
36.01
2.07
0.14
1.74
31.14
2.01
0.13
0.64
1.43
0.98

17.71

In 100 parts of dry matter

Total-N

Alb.-N

Ether-extract. . .
Soluble matter . .

Non-alb.-N

Ammonium-N. .
Organic base-N .

Other-N

'Total acid

Common salt . . .

5.64
1.00
5.21

77.74
4.64
0.43
2.39
2.12
4.07

49.37

5.79
1.00
5.13
80.76
4.79
0.43
2.04
2.26
4.02

5.79
0.80
5.10

82.53
5.18
0.70
1.97
3.09
2.73

48.95

5.76
0.38
4.84

86.49
5.57
0.36
1.79
3.98
2.72

49.17

' This sample contained the stomach, intestines, pyloric coecum and very
little liver.

xviii]

Congress of Applied Chemistry

271

2). The second sample* was prepared on the 3rd of Oct.
1911, and after 1, 14, and 53 days respectively, a portion was
taken for analysis: —

In 100 parts of fresh
sample

In 100 parts of dry
matter

Days after
prepara-
tion

Date of analysis .

Water

Dry matter

Total-N

Total acid

Ether-extract .

Soluble matter. . .

Non-alb.-N

Ammonium-N . . . ,
Organic base-N . . ,

Creatinine-N

Creatine-N

Xanthine base-N ,

Other-N

Natrium chloride . .

64.39
35.61
2.29
0.95
6.84
25.61
1.59
0.10
0.81
0.01
0.02

0.66
13.61

63.00

36.99

1.05
6.89

27.14
1.85
0.12
0.73

Trace

Trace
0.06
1.00

13.94

60.33

39.67

1.14

28.90
2.07
0.14
0.71
Trace
Trace
0.03
1.21

0
100,

2.
19.
71.

1
37.

0.0

100.0

2.85

18.61

73.27

4.99

0.32

1.96

Trace

0.16

2.71

37.62

53
0.0
100.0

2.88

72.86
5.21
0.35
1.79
Trace
Trace
0.08
3.05

The results of the above two analyses may be summarized as
follows: —

Soluble organic matter
Alb.-N
Non-alb.-N
Anunonium-N

Monoamino-N

Organic base-N

Creatine-N

Creatinine-N

Xanthinbase-N

EtheT'^ztract

Total acid

(1)

(2)

Gradually increased

Do.

" decreased

" increased

Do.

Increased at first and

Gradually increased

decreased hence-

forward.

Gradually increased

Do.

" decreased

Do.

Gradually decreased
11 11

Somewhat decreased

11 u

Do.

Decreased

Somewhat increased

> This sample contained more liver than the former one.

272 Original Communications: Eighth International [vol.

Thus the results of two analyses resemble each other in general
respects, only the contradictory results were observed with
ammonia and with total acid. This may be due to the differ-
ences of materials and also the temperature during the experi-
ments.

3). I will add here some quahtative tests made about the
distillates obtained by the steam distillation of two shiokara-
preparations, in neutral as well as in acid reaction.

10 days after

61 days after

In the distillate

preparation

Alcohol

i+y (very little)

(— )*

Aldehyde

(-)

(— )

Acetone

(-)

(— )

Indol

(-)

(— )

Phenol

(-)

(— )

Formic acid

(+) (trace)

(+) (distinct)

In the residue

Lactic acid

(+) (distinct)

Succinic acid

(-)

In the water extract of
the natural sample

Tryptophan

(+)

StTMMABT of the REStTLTS

1). Various samples, examined at different stages of ripening,
gave all acid reaction chiefly due to lactic acid.

2). Autolysis is going on in the freshly prepared "Shiokara,"
and decreases gradually as the ripening process proceeds.

3). The enzymes found in "Shiokara" are diastase, lipase,
and trypsin. The last one acts not only in weak alkaUne solu-
tion but also even in neutral or in faintly acid reaction.

4). Micro-organisms play also some important r61e during the
ripening process.

'(+), indicates presence; (— ) absence.

xviii] Congress of Applied Chemistry 273

5). Temperature has also great influence upon the action of
enzymes and microbes.

6). During the ripening process, the increase of soluble matter
non-albiuninoid nitrogen, especially monoamino-nitrogen, and
the decrease of protein, organic bases, creatine, creatinine, and
purin bases were observed.

In conclusion I express my thanks to Profs. CJ. Suzuki and
S. Machida for their kind advices given during the work.

QUANTITATIVE DETERMINATION OF CREATINE,

CREATININE AND MONOAMINO-ACIDS IN SOME

FISHES, MOLLUSCA AND CRUSTACEA

By Y. Okuda
College of Agriculture, Imperial University, Tokyo

I. Creatine and Creatinine

For the determination of creatine and creatinine, the flesh
freed from bones, heads, fins, scales, and internal organs* was
chopped and extracted with water at 50-55° for one hour. The
residue was treated two times more in the same way. The whole
extract was now boiled for a short time to remove most of the
proteins by coagulation and filtered. The filtrate was evapo-
rated under diminished pressure to a small volume and was
divided into two portions. One portion of it served directly for
the determination of creatinine after Fohn's colorimetric method,
while the other portion was previously boiled with nearly 4 per
cent, sulphuric acid for two hoxirs, to convert the creatine pres-
ent into creatinine, and after removing the sulphuric acid by
means of barium hydroxide, it was subjected to the determination
after Fohn. From the difference of these two determinations
we can calculate the quantity of creatine originally present in the
flesh, 1 mg creatinine being equivalent to 1.16 mg creatine.
The results obtained were as follows :

' The case of clam was exception, as its whole body was used.

275

276 Original Communications: Eighth International [vol.

Name

In 100 parts of fresh
substance

Water
g

Creatine
g

Creati-
nine
g

In 100 parts of
dry matter

Creatine

Creati-
nine
g

Bonito (Gymnonsarda aifinis

Cantor)

Timnyfish (Thunnus schlegeli

Steined)

"Katsuobushi" (Steamed and

dried bonito)

Salmon (Oncorhynchus tshawyt-

scha Walbaum)

Snapper (Pagrus major)

Carp (Cyprinus carpio L) . . . .

Shark

Lobster (Palinurus japonicus

Gray)

Crab (Neptmius pelagicus M-

Edw)

Cuttle-fish (Sepia esculentaHoyle)
"Kakisxn-ume" (Chopped and

dried cuttle-fish)

Clam (Cytherea meretrix L) . . .

72.165

72.402

14.808

63.300
77.340
79.160
79.800

79.920

84.500
81.699

27.570
90.490

0.649

0.497

0.453

0.560
0.754
0.421
0.655

Trace?

Trace?
Trace

Trace
Trace

0.134

0.064

0.660

0.067
0.070
0.077
0.134

Trace?

Trace?
Trace

Trace
Trace

2.011

1.800

0.531

1.525
3.327
2.020
3.242

0.481

0.232

0.775

0.182
0.308
0.369
0.663

The materials used for the determination were very fresh,
except the salted flesh of salmon.

We see from the above result that all of the examined fishes
contained comparatively much creatine and creatinine,' on the
contrary in mollusca and Crustacea, the existence of these two
compovmds was doubtful, at least, they must be present only
in traces. In fresh fish we found generally more creatine than
creatinine, while in dried bonito the reverse was observed. It is
therefore possible that a part of creatine is transformed into
creatinine during the preparation of the food.

' Van Hoogenhuyze and H. Herploegh found per kilogramm flesh of ox,
sheep, pig and horse 4.4, 4.1, 4.5 and 3.8 g creatine respectively. (Zeitschr.
f . physiol. Chem., 1905, 46, 432.)

xvin] Congress of Applied Chemistry 277

It may be mentioned here that the water extract of clam gives
only slight yellowish red coloration instantly after addition of
picric acid and soda after Folin, thus showing that only a trace
of creatinine is present in it, but after standing for many hours
at room temperature, it takes a dark red color. After some
tests we found that the glycogen, originally present in the ex-
tract, is gradually acted upon by diastatic ferments of clam
itself, and the sugar thus resulted may impart this red coloration.
The presence of diastatic ferment in the clarn is easily shown in
the usual way.

II. MONOAMINO-ACIDS

For the determination of monoamino-acids Sorensen's formol
titration method was adopted. Of course, this method does not
hold good for every monoamino-acid, but in the case of fish
flesh, the quantity of the amino-acids being very little, the
method of Van Slyke is not conveniently applied.

I have made some preliminary tests also, and found that
the presence of organic bases, like arginine, lysine, histidine,
etc., more or less interferes with the result of the formol method,
so it is better to remove these bases previously. But the pres-
ence of creatine has apparently no effect upon this determi-
nation.

150 g minced fresh flesh, free from bones, heads, fins, scales
and internal organs was extracted in the similar way as men-
tioned above and the aqueous extract was boiled and slightly
acidified with acetic acid to remove coagulable proteins, fiiltered,
neutralized and evaporated by a low pressure to a small volume,
acidified with sulphuric acid and precipitated with phospho-
tungstic acid in the usual way. The filtrate of the phospho-
tungstic precipitate, after the removal of the phosphotungstic
and sulphuric acid by means of barium hydroxide, was evapo-
rated, in neutral reaction, again to a small volume and titrated
according to the usual formol method. Thus the following
result was obtained:

278 Original Communications: Eighth International [vol.

Substance

Water

N of Mono-

amino-acids

ing.

Remarks

InlOOg
fresh
flesh

InlOOg
dry
flesh

Carp I

76.609
76.789
73.516
69.371
76.787
81.998
75.975
81.671

0.022
0.024
0.011
0.022
0.016
0.035
0.146
0.089

0.094
0.103
0.041
0.072
0.069
0.194
0.608
0.485

Tunny, bonito, porgy and cuttle fish
applied to the above determination
were fresh. Spiny lobster, Crussian
carp and Carp I were still Uving when
they were analyzed. Carp II was ana-
lyzed standing 50 hours after death
at room temperature (12°C). The
increase of monoamino-acids after that
time was very insignificant.

Carp II

Tunny

Bonito

Porerv

Crussian carp. .
Spiny lobster . . .
Cuttlefish

We see from the above resiilts that the contents of monoamino-
acids are generally very little in fish, while moUusca and Crus-
tacea contain a little more.

III. On Different Forms of Proteins in the Flesh of Fish

For this purpose, the flesh was extracted^ with water, alcohol,
NaCl, and KOH, respectively, and the quantity of total and
albuminoid nitrogen in each extract was determined according
to Kjeldahl's method.

• 10 g fresh flesh was extracted with 100 c.c. of solvent for 24 hours at 10°C.

xvin]

Congress of Applied Chemistry

279

In 100 g fresh

In 100 g dried

Sum of each

flesh

N as 100

Flesh

Solvent

Total

Prot.

Total

Prot.

Total

Prot.

Crussian carp

0.746

0.476

4.144

2.644

17.171

13.583

(Carrassius aurar

0.2%KOH

2.003

1.793

11.127

9.960

46.077

51.125

tusL)

70%Alcohol

0.386

0.174

2.144

0.966

8.S99

4.961

10% NaCl

1.212

1.064

6.732

5.910

27.881

30.339

Carp

0.479

0.326

2.182

1.485

11.129

10.326

(Cyprinus

0.2% KOH

1.715

1.341

7.812

6.108

39.809

43.477

carpio L)

70% Alcohol

0.492

0.240

2.241

1.093

11.420

7.602

10% NaCl

1.622

1.250

7.388

5.694

37.649

39.605

Spiny lobster

1.600

0.736

6.659

3.063

23.808

22.816

(Palinurus

0.2% KOH

2.138

1.212

8.898

5.044

31.813

37.572

japonicus Gray)

70% Alcohol

0.934

0.119

3.887

0.495

13.898

3.689

10 % NaCl

2.048

1.158

8.544

4.819

30.474

35.898

Cuttle fish

H,0

0.932

0.351

5.085

1.915

19.984

(Sepia esculenta

0.2% KOH

1.775

1.223

9.864

6.673

38.059

Hoyle)

70% Alcohol

0.602

—

3.285

—

12.908

10% NaCl

1.354

7.387

—

29.032

The amount of proteins extracted by alkali was generally
much greater than that extracted by other solvents. The pro-
teins soluble in 10% NaCl, as globulins, were also much, water
soluble proteins as proteose and albumine not much and the
proteins as prolamins very little.

IV. Form of Nitrogen in Some Marine Animals
The analytical results are shown in the following table:

Summaries

1. All of the examined kinds of fish contained comparatively
much creatine and creatinine, but the flesh of moUusca only
trace, in the flesh of Crustacea the existence of these compounds

280 Original Communications: Eighth International [vol.

was doubtful. The quantity of creatine was generally much
more than that of creatinine, in all fresh fishes.

2. In all marine animals examined the quantity of organic
base nitrogen is much more than that of monoamino-acid nitro-
gen, and the amount of the latter is generally very little in fish,
but somewhat much in lobster and cuttle fish.

3. Most of proteins are soluble in dilute alkali solution, the
proteins soluble in 10 per cent NaCl were also much, this fact
must be cared on the preservation of fish.

The experiments have been made by the writer under the
direction of Professor Dr. U. Suzuki, and it is my pleasant
duty to thank him for his kind advices given during the progress
of the work.

XVIll]

Congress of Applied Chemistry

281

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THE EFFECT OF MODIFYING THE GLUTEN SUR-
ROUNDING OF FLOUR

Geo. a. Olson
Pullman, Wash.

The results included in this article are a part of our cereal investi-
gation work. This particular article deals with the modifying of
the gluten surroundings in flour with a view of searching for the
causes which affect the quality of the flour for baking purposes.

The oldest idea is that the gluten in flour holds the gas in panary
fermentation. The capabilities to distend depends upon the
physical qualities of gluten. Wood believes the quantity of the
gluten is modified according to the amount of salts and acid
present, which in turn influences the shape of the loaf, and treats
this subject fully in his article on " The Chemistry of the Strength
of Wheat Flour." *

The writer in the following experiments studied the effect of
adding directly definite amounts of acid, alkali, or salt to original
dialyzed, decanted, dough, gliadin-free and nitrogen-free with
gliadin added flours. The nitrogen components of treated and
untreated flour have also been included. The quantity of dry
gluten and the nitrogen in the gluten of these respective flours
was determined in each case.

In order to remove the salts from a flour by either dialysis
or decantation, it is obvious that the flour must be soaked in
distilled water and then dialyzed or allowed to stand and the
supernatant liquid removed after the flour has settled to the
bottom of the jars. This method at once brought up the question
as to whether or not a flour could be water soaked and remilled
into flour again. The first trial with dialyzed flour, however,
determined that such a procedure was possible.

Perhaps the greatest difficulty experienced in this undertaking
was the drying of the water-soaked flour. Aside from avoiding

• Joum. Agr. So. Vol. 2, No. 2, Apr. 1907, p. 139, ibid. Vol. 2, No. 3, Deo.
1907, p. 267.

283

284 Original Communications: Eighth International [vol.

too high temperatures, in the drying process, the possibilities
for fermentation were most favorable and in consequence the
flour while drying required the strictest attention of both an
assistant and myself. The tendencies for fermentation were
particularly noticeable in the flour which had been dialyzed,
in spite of the fact that they had been treated frequently with
small quantities of chloroform.

In addition to recovering the water-soaked floiu-, the dialysate
and water decantations were saved and either reduced to small
volume or evaporated to dryness.

The method of preparing the dialyzed, decanted, and dough
flours are described in the following paragraphs :

Dialyzed Flour. Approximately kilo lots of flour were thoroly
worked into paste with water and then dialyzed in a cool room,
frequently changing the dialysate for a fresh supply of distilled
water. Suflicient chloroform was used to check fermentation.
After a course of three days the dialysis was considered complete.
The colloidal material was then thoroly stirred and poured
thinly over glass window panes and allowed to dry (frequently
stirring) at a low temperature. When it solidified into lumpy
masses all danger of fermentation was apparently removed. The
lumpy masses were then worked out into thin layers and allowed
to dry over night without any attention. The following morning
these thin layers of flour crumbled readily and were then ground
into small pieces about the size of clover seed, when it was dried
further. When satisfactorily dry the work was considered com-
plete after milUng and finally bolting thru a lOxx bolting cloth.

Decanted Flour. Kilo lots of flour were worked into paste
and then diluted with four liters of distilled water for every kilo
of flour; after settling, the supernatant Uquid was removed, and
the operation frequently repeated for a period of three days, after
which the flour was finally dried in the same manner as described
for the dialyzed flour. Sufficient chloroform was used throughout
the experiments to allay fermentation.

Dough Flour. Not knowing what influence the water would
have upon the physical properties of the flour, it was necessary,
in order to obtain checks, to treat the flour with enough water
to make a dough, allowing it to stand for three to four hours.

xviii] Congress of Applied Chemistry 285

then rolling this dough out into thin sheets, crumbhng and mill-
ing as described for the dialyzed flour.

Plan of Investigation. After the treated flours were prepared
as described above, the next step consisted in planning a method
whereby the physical properties of the glutens could be studied.
After some consideration and thought it was decided that in
place of taking bits of gluten prepared from these flours and
subjecting them to acid, alkali or salt as Wood did, that the
flour be treated with these agents directly and the changes occur-
ing, if any, be noted, and the amount of dry gluten and the
nitrogen in the gluten be determined. It can readily be seen
that such a plan takes into account other things besides salts and
acids, since it would also determine the effect of adding these
reagents to the flour directly. Later baking tests of flour treated
similarly could be made and the influence of the reagents upon the
shape of the loaf be noted.

Gluten Determinations. Ten gram lots of the treated and origi-
nal flour were mixed with six cubic centimeters of either dis-
tilled water or N-10 normal hydrochloric acid, sulfuric acid,
phosphoric acid, sodium hydroxide, potassium hydroxide,
dipotassium acid phosphate, disodium acid phosphate, dicalcium
acid phosphate, sodium chloride, sodium sulfate, aluminum sul-
fate and magnesium sulfate, and worked up into small wads.
These wads of flour were then allowed to stand for one hour, after
which they were finally washed over silk with running water.
In some cases the particles of gluten cohered and were easy to
gather, while in other instances the gluten particles scattered and
fell upon the silk. All scattering glutens which fell upon the
silk, together with those that cohered, were gathered together,
washed, dried and weighed according to the usual method. The
length of time required for drying was 20 hours at maximum
temperature of the water oven. Nitrogen determinations of the
dry glutens were finally made by the Kjeldahl method. The
results of the percent of gluten, weight and percent of nitrogen
in the gluten, percent of total nitrogen and percent of total nitro-
gen calculated from the nitrogen in the gluten from the original
flour as 100, are given in the following table, which is subdivided
into four. separate parts according to the treatment of the flour.

286 Original Communications: Eighth International [vol.

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xvm] Congress of Applied Chemistry 287

Comparing the data obtained for dry gluten in the original dough,
dialyzed, and decanted flours as modified by either water, acid,
alkali, or salt, it will be noted that with the exception of sodium
sulfate none of the N/lO solutions of salt, acid, or alkali gave as
high results as was obtained with the water in the original flour.
In Table 1 (a) the prejudicial* influence upon the gluten increased
in the order named, disodium acid phosphate, sodium chloride,
magnesium sulphate, dipotassium acid phosphate, dicalcium
acid phosphate, aluminum sulfate, sodium hydroxide, potassium
hydroxide, sulfuric acid, phosphoric acid, and hydrochloric
acid.

It will be further noted from the results given in table 1 (b)
that the dough flour agrees fairly well with the original flour, the
widest variations occurring where the flour had been treated with
sulfuric acid, phosphoric acid, and hydrochloric acid. In the
same way the weight of nitrogen varied and the percent of total
nitrogen calculated on the basis of 100 for the original flour per-
haps illustrates more clearly the effect of the acid upon the
dough flour when compared with the original.

In regard to the dialyzed flour, when compared with the origi-
nal, water has affected the yield of gluten and the weight of
nitrogen to a slight extent. Sulfuric acid, phosphoric acid and
hydrochloric acid have been prejudicial even to a much greater
extent than was the case in the dough flour. Sodimn hydroxide
and potassium hydrocide were apparently beneficial,** the former
more so than the latter. Comparing salts of phosphoric acid,
the sodium salt increased the gluten and nitrogen content to a
greater extent than the potassium salt, while the calcium salt
was slightly prejudicial in this case, being an exception to that
observed in case of the dough flour. Sodium chloride was bene-
ficial when weight of the nitrogen contained in the gluten is
considered. On the basis of weight of nitrogen, alimiinum
sulfate was prejudicial, sodium sulfate was without effect and
magnesium sulfate was beneficial. Comparing the influence of

* Prejudicial refers to decreased amounts of dry gluten or nitrogen in the
gluten.

** Beneficial refers to an increased amount of dry gluten or nitrogen in the
gluten.

288 Original Communications: Eighth International [vol.

the reagents on the gluten content and weight of nitrogen with
water, it will be noted that disodium acid phosphate, magnesium
sulfate, sodium sulfate and sodium chloride were beneficial in
the order named, while dipotassium acid phosphate, dicalcium
acid phosphate, sodium hydroxide, potassium hydroxide, alum-
inum sulfate, phosphoric acid, sulfuric acid and hydrochloric acid
were prejudicial in the order named.

The dalysate from dialyzed flour (based upon the weight of
flour used) contained 0.655 percent total solids, of which 0.57
per cent was combustible and 0.084 percent was ash.

The flour obtained after decanting the soluble extract gave
the most remarkable results of any in the series. Neither water,
salts, nor acid yielded gluten, while sodium and potassium hydrox-
ides were beneficial to gluten formation. In addition, calcium
hydroxide, glycerol, alcohol and flour extract were tried and
only the glycerol and calcium hydroxide were found to be bene-
ficial.

Collectively, tlie results given in Table I (a, b, c and d) clearly
show that acid is more prejudicial than alkali of same normal
strength and the salts with one exception (aluminum sulfate in
case of the dialyzed flour) has practically a very slight effect upon
the physical properties of the gluten. On this basis, sodium was
least active and calcium most active on gluten disintegration.
There is no doubt but that the substances contained in the
water extract play an important part in modifying the physical
properties of the gluten. Whether the substances contained
in this extract, which play so important a role, are inorganic or
not must be determined. It appears that the (OH) radical tends
to produce coherence. This view is supported from the results
obtained in the dialyzed flour when compared with the original,
and in the decanted flour where other substances failed to bring
on coherence. Just what causes the disintegration of the gluten
complex has not as yet been satisfactorily established.

Physical Condition of the Gluten. The gluten in the flour
studied from physical appearances was fair. None of the salts
used showed any marked variation in the physical appearances
of the gluten other than was observed in the case of water.
N/10 acids tended to produce scattering glutens, which when

xvni]

Congress of Applied Chemistry

289

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290 Original Communications: Eighth International [vol.

gathered were dry and seemingly free from water, while alkalies
of the same strength affected the gluten in such a way that in
the process of washing out the starch the gluten rolled or washed
down with the water on to the silk in little bits, which when col-
lected together, (the gluten) had a very dry touch, indicating that
glutens treated in this way contain very little water. The
amount of water contained in N/10 acid and alkali prepared
glutens was not determined.

Influence of Acids, Alkalies and Salts on Gluten from Patent
Flours. Four patent flours were next studied with the object of
determining whether or not the addition of acid, alkali, or salt
to the flour modified the physical properties, the yield and the
nitrogen content of the gluten in these flours. The flour selected
for this series of investigations represent four different localities
with different climates, soil and types of wheat. The glutens
from these flours varied considerably in their physical qualities.

As in the case of the previous experiments, ten gram samples
were mixed with six cubic centimeters of either N/lO acid,
alkali or salt, and in addition to these Uke qualities of N/lOO
and N/l acid and alkali were tried. These different lots were
worked up into wads and the gluten collected by washing over
silk. The weight and the per cent of the nitrogen in the gluten
are recorded in Table II. The grams of nitrogen in the gluten
and the ratio of total nitrogen in the flour to nitrogen in the
gluten are recorded in Table III. In place of phosphoric acid,
lactic acid was used in these experiments. Potassium hydroxide
was not tried.

It will be noted from the data given in Table II that the acids,
alkali and salt affected the weight of gluten similarly as found in
Table I (a). "While the strength of N/IOO acid was sUghtly
prejudicial, increasing the strength to N/lO and N/1 had
marked effects upon the yield of gluten when compared to those
treated with distilled water and run as controls. The increased
or equal weight of gluten in the flour treated with N/l sulfuric
acid compared to N/10 sulfuric acid was due to the fact that
the former was poorer in nitrogen. Disodium acid phosphate
was beneficial in all cases. Sodium chloride and sodium sulfate
were beneficial in three out of four instances. It was impossible

xvin]

Congress of Applied Chemistry

291

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292 Original Communications: Eighth International [vol.

to obtain gluten from flour treated with N/1 sodium hydroxide,
N/lOO sodium hydroxide was less prejudicial than N/10. The
most pecuhar modification of gluten that took place was with
N/lOO sodium hydroxide in case of the Bridgeport bluestem
where the weight of the gluten increased. The per cents of
nitrogen in the glutens varied irregularly.

The data given in Table III show the actual amounts of
nitrogen entering into the gluten make-up. The ratio of total
nitrogen in the flour to that found in the gluten shows how re-
agents of identical normal strengths modify the nitrogen com-
plexes in the gluten. Particular attention should be called to
the effect of N/lOO acid. The ratio constant was remarkably
uniform for each regardless of source or natiu-e of the flour.
SUghtly higher ratios were obtained with hydrochloric acid
and sulphuric acid than with lactic acid. With N/10 solution,
sulphuric acid was the least prejudicial. Hydrochloric acid
disintegrated the gluten rapidly at this strength, while with
lactic acid practically no gluten was obtained. No results could
be obtained with either N/1 hydrochloric acid or N/1 lactic
acid. The accompanying cut illustrated the effect of N/lOO,
N/10 and N/l sulphxu-ic and hydrochloric acids upon the yield
of gluten.
(CUT) (1) (See legend on back of photo.)

The physical condition of the glutens resulting from washing
the floiu- after it had been treated with hydrochloric acid, lactic
acid, and sodium hydroxide of N/lOO, N/lO and N/l strengths
are tabulated in Table IV.

From the physical appearances of the gluten as affected by
either acid or alkaU, it may be said that alkali behaves similarly
to acid of the same normal strength.

Flour with Gliadin Removed. Since it is more difficult to
remove glutenin than gliadin from flour it was decided to study
the influence of acid, alkali, and salts on flour with the gliadin
removed. Flour was repeatedly extracted with cold 70 per cent
alcohol and finally dried, milled and bolted thru a lOxx bolting
cloth. As in previous experiments, 10 grams of flour and 6
cubic centimeters of water or specified reagent were used. The
amount of gluten obtained with water 0.14%, N/lO sodium

XVIIl]

Congress of Applied Chemistry

293

TABLE IV

Physical Properties of Gluten as Affected by Acid and
Sodium Hydroxide

HCl

H,SO.

CiH,0.

Na (OH)

N/1

disintegrated

soft, massive,
lack cohe-
siveness.

disintegrated

tough dough,
soapy and diffi-
cult to remove
gluten

N/10

separated in par-

same as for

disintegrated

separates into par-

ticles, firm and

N/10 HCl

ticles, firm and

seemingly dry

dry

N/100

excellent cohe-

same as for

soft, voluminous

sion, elastic

N/100 HCl

gluten, easy to
wash, elastic

hydroxide 0.712%, N/lO sodium chloride 0.100% and N/lO
hydrochloric acid none, was insignificant when compared with
flours treated by dialysis or decantation and as a result it was
considered futile to try other reagents on this gliadin-free flour.
These results, however, appear to indicate that gliadin does not
act as an acid, alkali, or salt, for if such was the case, it is reason-
able to believe that all of the glutenin could have been recovered
by the addition of an acid, alkali, or salt.

Nitrogen-free Flour with Gliadin Added. In the previous
paragraph, the effect of acid, alkaU, and salt upon gliadin-free
flour was discussed. In a similar way the same original flour
was treated with weak potassium hydroxide N/50 several times.
The supernatant liquid resulting after the flour stood for four
hour intervals was replaced with fresh lots of potassium hydroxide
and finally the treated flour was washed with frequent changes of
distilled water, then rolled into thin layers, dried and milled.
The amount of nitrogen components present after the above
treatment was determined and found to be a trace of globulin
and albumin nitrogen, amid nitrogen 0.119%, alcohol soluble
(gliadin) nitrogen 0.133%, total nitrogen 0.301%. The total
nitrogen in the original flour was 1.63%.

294 Original Communications: Eighth International [vol.

It appears from these results that water is more prejudicial
than either N/lOO hydrochloric acid or sodium hydroxide and
N/IO hydrochloric acid or sodium chloride. It is reasonable
to believe, however, that such was not the case, since the gliadin
in the course of preparation and purifying has been altered and
when dried and ground is made up of minute hard particles of
ghadin. Just as dried gluten takes up water only after long
periods of soaking, so it is with these hard particles of gliadin
which, when mixed with water, require long periods of time to
even swell. Thus when flour is incorporated with an addition
of these hard particles of gUadin and then mixed into wads with
water and gluten determinations made, the finest particles of
gliadin pass thru the fine silk and are lost. The only gliadin that
can be recovered are the partly swelled particles which are too
large to pass thru the silk. The presence of N/lOO hydrochloric
acid or sodium hydroxide and N/10 hydrochloric acid and sodium
chloride, undoubtedly, has increased the tendencies of the hard
particles of gUadin to absorb more water than was the case with
distilled water and as a result increased the number of particles
too large to pass thru the silk hence a larger yield. With larger
quantities of water than was used in these experiments more
gliadin would undoubtedly have resulted since gUadin takes up
water in time and with an increased amount accelerates water
absorption thereby increasing the number of larger particles.

Nitrogen Components of Treated and Untreated Flour. Aside
from studying the effect of acid, alkali and salts upon the yield
of gluten and the weight of nitrogen in the same, a systematic
study of the nitrogenous components in the flours mentioned above
was made. The first flours investigated were the dough, dialyzed,
and decanted compared with the original. All nitrogen deter-
minations were made according to the straight Kjeldahl method.
The total nitrogen, the percent of nitrogen of flour entering into
the gluten makeup, the percent of alcohol soluble nitrogen (using
70% alcohol by volume), the percent of nitrogen compounds
precipitated by phosphotimgstic acid from a one percent sodium
chloride soluble of flour and the percent of nitrogen in the salt
soluble extract not precipitated by phosphotungstic acid were
determined.

xvni]

Congress of Applied Chemistry

295

TABLE V
Nitrogen Components in Differently Treated Flovirs

Nitrogen
in Flour

Per Cent.

Nitrogen in
Gluten

Per Cent.

Gliadin
Nitrogen

Per Cent.

Edestin

and
Leuoosm
Nitrogen
Per Cent.

Amide
Nitrogen

Per Cent.

Glutenin
Nitrogen
baaed on

Flour
Per Cent.

Ghitenin
Nitrogen
baaed on
Gluten
Per Cent.

Original.
Dough . . .
Dialyzed .
Decanted

1.63
1.63
1.69
1.66

1.5218
1.5176
1.4658
1.4255*

0.602
0.609
0.6^3
0.756

0.378

0.056

0.594

0.4858

0.308
0.273

0.070
0.077

0.689
0.554

0.4648
0.3195

The results given in Table V show a higher nitrogen content
for the dialyzed and decanted flours, than was found in case of
dough and original flour. These differences in total nitrogen
in the dialyzed and decanted flours may be due to the renaoval
of nitrogen-free or nitrogen-poor material. The fact that
the decanted flour contains less nitrogen than the dialyzed flour
would tend to indicate that this was not the case; on the other
hand, it must be considered that small quantities of the more
highly complex nitrogen bodies are carried away with water
and with the large quantities of water used repeatedly for de-
cantation some of the higher nitrogen bodies have been affected
and their losses have resulted in diminishing the nitrogen con-
tent. The amount of nitrogen in the glutens (expressed in per-
cent of the nitrogen of the flour in gluten), also supports this
thought, being higher in amounts for the original and dough
than in either the dialyzed or decanted. Although, it was im-
possible to obtain any gluten with water (see Table 1, d) in the
decanted flour, the result 1.4295% nitrogen cannot be far off
from the actual amount present if a gluten could have been
made. This result has been calculated from the data given in
table 1, a & d, and confirmed by those given in Table V. The
increased amount of alcohol soluble nitrogen in the decanted
flour cannot be entirely accounted for by the removal of the
nitrogen-free and nitrogen-poor material. In this same flour
there is less precipitated nitrogen and more nitrogen not pre-
cipitated by phosphotungstic acid. The same is true to a less

* Calculated.

296 Original Communications: Eighth International [vol.

extent in case of the dialyzed flour. The glutenin was obtained
by difference between the nitrogen obtained in gluten and the
sum of alcohol and the salt soluble. Another column for glutenin
has been inserted for the benefit of those who subtract the alcohol
and salt soluble from the total nitrogen.

In Table VI the results given in Table V are calciilated as
proteins using the factor 6.25.

TABLE VI
Nitrogen Components Calculated as Proteins

Protein m
Flour

Per Cent.

GHadin
Per Cent.

Edeetin

and
Leucosin
Per Cent.

Amides
Per Cent.

Glutenin
baaed on

Flour
Per Cent.

Glutenin

based on

Gluten

Per Cent.

Gluten
Per Cent.

Original .
Dough . . .
Dialyzed .
Decanted

10.19
10.19
10.56
10.38

3.76
3.81
3.89
4.73

2.36

1.93
1.70

0.36

0.43
0.49

3.71

4.31
3.46

3.04

2.91
2.00

9.51
9.49
9.16
8.91

Owing to the increased amount of alcohol-soluble nitrogen
obtained in the decanted floiu- over that found in the original,
two other flours were treated by the decantation method. The
nitrogen components of these flours in conjunction with the same
flours imtreated were also made. The data for the original and
decanted flour given in Table V are included in Table VII for
comparison.

TABLE VII
Nitrogen Components in Patent Flour Compared with the Same Flour after

Decantation.

Total
Nitrogen

Per Cent.

Gluten
Nitrogen

Per Cent.

Gliadin

Nitrogen

Per Cent.

Edeslin
and ^
Leucosin
Nitrogen
Per Cent.

Amide
Nitrogen

Per Cent.

Glutenin
Nitrogen
based on

Flour
Per Cent.

Glutenin
Vitrogen
based on
Gluten
Per Cent.

Pullman, Original. . .

Decanted .

Bridgeport, Original . .

Decanted
Vancouver, Original . .

Decanted

1.18
1.20
1.79
1.74
1.63
1.66

0.987

1.526

1.5218
1.4255*

0.651
0.686
0.910
0.950
0.602
0.756

0.329
0.217
0.350
0.245
0.378
0.273

0.028
0.021
0.049
0.035
0-.0S6
0.077

0.172
0.256
0.481
0.510
0.594
0.554

None

0.217
0.486

* Calculated.

Increasing the strength of acid alters the physical quaUty of gluten.

XVIIl]

Congress of Applied Chemistry

297

It will be seen from the results given in Table VII that the
amount of alcohol-soluble nitrogen in the decanted flours were
again found to be higher than that found in the original flours.
The amount of amid nitrogen foimd was, however, the reverse
of that found in Table V. When the nitrogen in the gluten
makeup is assumed to be composed of the ghadin and glutenin
it will be noted that the Pullman flour is devoid of glutenin.
On the other hand, if the total nitrogen is considered instead of
the nitrogen entering into the gluten makeup there is found
0.172% of gluten.

Extract from Decanted Flour. In addition to having determined
the nitrogen components of both the control and the decanted
flours, the extract of the decanted flour was also studied in regard
to the total nitrogen, nitrogen precipitated and not precipitated
by phosphotungstic acid. The total soUds and total ash were
also made. These results are recorded in Table VIII.

TABLE VIII
Nitrogen Components in Decanted Liquid from Patent Flour

Total Nitrogen
Percent.

Edeatin and
Lewcosin
Nitrogen
Percent.

Amide Nitrogen
Percent.

Total Asli
Percent.

Total Solida
Percent.

Pullman

Bridgeport

0.094
0.081

0.079
0.074

0.015
0.007

0.36
0.38

3.00
2.74

The results given in Table VIII show that the total solids and
nitrogen components for the Bridgeport flour are slightly lower
than was the case in the Pullman flour. It may be of interest to
know what these extracted substances are, since all or some one
of them may be important in determining the baking qualities
of a flour. Although some work has been done by the writer
along this line of research, nothing definite has been determined
thus far.

In Table IX an attempt has been made to correlate some of the
data given in preceding tables. The ratio of total nitrogen to
gluten nitrogen, as affected by water, sodium hydroxide, disodium

298 Original Communications: Eighth International [vol.

acid phosphate, and sodium sulphate on the one hand, with the
ratio of gliadin to glutenin nitrogen on the other, are recorded.

TABLE IX

Ratio of Total Nitrogen to Gluten Nitrogen as Affected by Alkali,

Water and Salt and Ratio of Gliadin to

Glutenin Nitrogen

Ratio of
gliadin to-
gluten ni-
trogen

Total nitrogen to gluten nitrogen ratio

N/10
Na (OH)

Water

N/10
Na.H2
(P0.)=

N/10
Naj
(SO.)

Pullman. .
Bridgeport
Vancouver

1:0.264
1:0.628
1:0.986

1:0.57

1:0.636

1:0.743

1:0.837
1:0.852
1:0.934

1:0.87
1:0.88
1:0.91

1:0.87
1:0.88
1:0.95

When flour was treated with either water, alkaU or salt the
ratio of total nitrogen to gluten nitrogen decreased as the ratio
of gliadin nitrogen to glutenin nitrogen decreased; this fact is
clearly brought out in table IX. In other words, the more
glutenin nitrogen a flour contains the higher will the gluten
nitrogen be. On the other hand, this view is contradicted, since
the flour which had the gliadin removed practically yielded no
gluten when treated with acid, alkali, or salt. In the same way
decanted flour having nearly all of its glutenin and gliadin
yielded only a part of its gluten when treated with sodium hydrox-
ide, potassium hydroxide, calcium hydroxide, and glycerol.
From these observations there appears to be some as yet unknown
substance or physical change which is more important in caus-
ing a transformation of the physical properties of gluten than
either acid, alkali, or salt. Whether this be in the form of some
organic salt, acid, or alkali, or not is a problem for the future.

XYOi] Congress of Applied Chemistry 299

Conclusions

1. Flours were either made into dough with water, dialyzed
or decanted, then dried, remilled and bolted into flour again.
Gluten determinations were made, using different reagents in
order to note the differences in yield caused by modifying the
surroundings.

2. Mixing untreated flour with N/lO solutions of different
salts, acids and alkali was prejudicial to the yield of gluten.
The prejudicial influence increased in the following order: Sod-
ium phosphate, sodium chloride, magnesium sulphate, potassium
phosphate, calcium phosphate, aluminum sulphate, sodium
hydroxide, potassium hydroxide, sulphuric acid, phosphoric
acid and hydrochloric acid.

3. Flour treated with sufficient water to form a dough then
dried and remilled into flour again, has resulted in slightly modi-
fying the gluten of that flour when compared with the original.
N/lO solutions of sulphuric acid, phosphoric acid and hydro-
chloric acid were more prejudicial to gluten formation in dough
flour than in the original one.

4. The gluten from flour which has been dialyzed has been
affected to a greater extent than was the case with the dough
flour.

5. No gluten could be obtained from the residual flour by
decantation when mixed with either water, salts or acids. Mixed
with N/10 solutions of sodium, potassium, and calcium hydrox-
ides and glycerol, varying amounts of gluten were obtained.
The amount of gluten obtained decreased with the hydroxides
used in the order mentioned above in No. 2 of conclusions.

6. N/10 solutions of salts appear to have no effect upon the
physical appearance of the resulting gluten when compared
with similar ones which were mixed with water. N/lO acid and
alkah tend to produce scattering glutens which when gathered
appear to be rather free from water. The amount of water
held by such glutens was not determined.

7. Using patent flours from various sources it was found that
the glutens prepared from these behaved similarly to those pre-
viously obtained and treated in Uke manner.

300 Original Communications: Eighth International [vol.

8. Patent flours were treated with N/100, N/iO, and N/1
sulphuric acid, hydrochloric acid, lactic acid and sodium hydrox-
ide. N/100 strength solutions had the least efifect upon the
yield of gluten. N/lO strength solutions, the hydrochloric
acid yielded some and the lactic acid practically no gluten.
Using N/1 strength solutions non-cohering gluten resulted with
sulphuric acid and no yield with hydrochloric acid and lactic acid.

9. In determining the nitrogen present in the glutens resulting
from treating flour with N/lOO strength, solutions of sulphuric
acid, hydrochloric acid or lactic acid, gave fairly concordant
results when compared with one another and the ratio of total
nitrogen to gluten nitrogen was practically constant, regardless
of the kind of acid used.

10. Flour with the gliadin removed does not form gluten
either in the presence of water, acid, alkali or salt. This fact
indicates that gliadin does not behave either as an acid, alkali
or salt.

11. Nitrogen free flour with gliadin added to it does not
form gluten either in the presence of acid, alkali or salt. The
failure to recover the admixed, previously dried, gliadin is
undoubtedly due to the limited time the gliadin was exposed to
water; in addition to the small quantity of water used.

12. In studying the nitrogen components of the original
dough, dialyzed, and decanted flam's, it was found that the
dialyzed and decanted flours showed slightly higher total nitro-
gen and alcohol soluble nitrogen contents than was found in
either the original or dough flour.

13. An attempt was made to correlate the ratio of gliadin
nitrogen to glutenin nitrogen with the ratio of total nitrogen to
gluten nitrogen as affected by either water, sodimn hydroxide,
sodium phosphate and sodium sulphate. According to the data
obtained it appears that the ratio of total nitrogen to gluten
nitrogen decreased as the ratio of gliadin nitrogen to glutenin
nitrogen decreased. On the other hand, it must be considered
that gliadin-free flour yielded no gluten and decanted flour jdelded
only a part of its gluten. Accordingly there appears to be as
yet some unknown substance which is important in causing a
transformation of the physical properties of the gluten.

A METHOD FOR THE DETECTION OF COLOR IN TEA

By E. Albebta Read, Ph.D., M.D.

Bureau of Chemistry, Department of Agriculture, Washington,

D. C.

The following method was devised for the purpose of detecting
color on tea, the United States Treasury Department having
issued regulations prohibiting the entry into this country of
colored teas.

For the demonstration of color and facing on tea, the chemical
methods, as suggested by Allen (1), Leach (2), Villiers et Collin
(3), and The International Committee (4), have usually been
employed. The difficulty with such methods is in the small
amount of color used and the masking of the color reactions by
the solution of natural color in the tea.

The method suggested in this paper has the advantage in that
it can detect much smaller amounts than can be found by chemi-
cal methods, but at the same time overlooks traces of color which
would be found by a compound microscope.

Hilger and Mayrhofer (5) suggest a method of rubbing wet tea
leaves on white paper, to detect artificial color.

The method suggested in this paper has an advantage over the
use of wet tea leaves in that a larger sample can be used in a
single examination, and it is more easily and quickly handled.
In cases where there is a blending of colored and uncolored tea,
many of the wet leaves might be used without detecting the
color. These authors also suggest the sifting of the tea, but no

(1) Commercial Organic Analysis, 1911; V, 658.

(2) Food Inspection and Analysis, 1911; 375.

(3) Traitd des Alterations et Falsifications des Substances Alimentaires,
1900; 258.

(4) Report on the Unification of Analytical Methods for Food-products;
1912; 148.

(5) Vereinbanmgen zur Einheitlichen Untersuchung und Beurtheilung von
Nahrungs-und Genusamitteln fur das Deutsche Reich, 1-3, 1897-1902; 54.

301

302 Original Communications: Eighth International [vol.

mention is made in the article of crushing the particles on paper
to demonstrate the presence of color, and thus not only affording
a method surer and safer for the chemist but also one which can
be used by mfen untrained in Science; therefore, making it avail-
able for tea examiners at the ports and for the tea tasters employed
by the importers and dealers in tea. It also allows the accurate
handling of large numbers of samples within a short time, thus
preventing the detention of tea at the ports for any considerable
time and consequent financial loss to the importer.

The articles needed for testing the tea are sieves, 16 to 24
meshes to the centimeter, a spatula or case knife and a piece of
unglazed, white paper.

A small amount of tea, about 25 to 50 grams, is placed in a
sieve and shaken over a piece of white paper. If the tea is
tightly rolled, it should be slightly crushed, either before putting
into the sieve or by rubbing it against the sieve. The dust on the
paper is then crushed by dragging over it a spatula or case knife,
pressure being applied by the finger to the end of the spatula.
This crushes not only the tea dust, but any particles of color which
are present. The process of dragging the knife across the paper,
streaks the color, making it more easily seen. A lens with a
magnification of 8 to 12 diameters is useful in detecting the
smaller streaks. Sunlight is desirable; bright light is essential
for this work.

This method will detect any coloring as blue, turmeric or car-
bon. An apphcation of the method has been made by Mr. G.
F. Mitchell, Supervising Tea Examiner, Treasury Department,
to the detection of facing on tea. Black, unglazed paper is used
in place of the white paper. The facing leaves a white streak on
the black paper.

Microchemical Tests for Color:

A black streak would suggest carbon; the blue may be Prussian
blue, indigo or ultramarine; and a yellow streak suggests turmeric.
These may be identified as follows: The carbon, by its glossy
appearance; the blue and turmeric can be tested directly on the
paper or by mounting on a microscopic slide. To the blue
streak on the paper or to the particle on the microscopic slide, add
a drop of 40 % sodium hydroxide. Prussian blue will turn

xvra] Congress of Applied Chemistry 303

yellowish-brown; indigo or ultramarine will remain unchanged
in color. Ultramarine is discolored by acid; indigo remains
unchanged when treated with either acid or alkali. Turmeric
turns bright red when a drop of a mixture of equal parts of boracic
acid and concentrated hydrochloric are added to the yellow streak.
Concentrated sulphuric also turns turmeric bright red.

RECHERCHE DE PETITES QUANTITES DE GRAISSE
DE COCO DANS LE BEURRE DE VACHE

Par M. Lucien Robin
Chimiste au Laboratoire Municipal de Paris, Paris, France

La m^thode d'analyse du beurre que j 'expose dans iin premier
memoirs, est assez sensible pour d^celer 10% de coco. En y
apportant quelques variantes, on pourrait en augmenter encore
la sensibility, ainsi que j'ai pu le verifier d^jk par un certain
nombre d'exp^riences que je tiens k renouveler encore, et dont
les r^sultats me semblent assez int^ressants pour que je les
fasse connaltre.

Le beurre de coco renfermant beaucoup d'acides caprique,
caproSque et laurique dont la majeure partie se trouve dans le
groupe des acides gras insolubles dans I'eau, et ces acides ayant
un indice de saponification 6lev6, j'avais pens6 qu'en isolant
assez d'acides gras de ce groupe pour en determiner cet indice,
je trouverais peut-6tre 1^ un renseignement pr^cieux.

Un certain nombre de tentatives de ce genre m'ont bien fait
voir que I'indice de saponification des acides de ce groupe obtenus
avec le coco, 6tait relativement 61ev6, et le plus souvent sup6-
rieur k celui que donnent les mSmes acides retires du beurre
pur, mais que pourtant on ne pouvait esp6rer pouvoir s'en tenir
exclusivement k cette determination pour aflBrmer la pr&ence
de 5% de coco.

Mais il n'en est plus de mime, si Ton compare les rapports
6tablis comme je vais I'indiquer:

Appelons I S I'indice de saponification du beurre

i s celui de ses acides gras insolubles dans I'eau
I E les acides gras insolubles dans I'eau dos& sui-

vant ma m^thode (ler m^moire).
S E les acides solubles dans I'eau dos^s aussi sui-
vaat ma m^thode.
305

306 Original Communications: Eighth International [vol.

Si Ton 4tablit les rapports suivants:

g^ = Riet j^ =R2

on constate que les beurres purs doanent

B} toujours inf^rieur k BP

tandis que les beurres cocotfe k 5% seulement donnent le con-
traire : R^ sup&ieur k R^

si I'on ajoute 10% d'um melange k parties ^gales de coco et de
margarine, R^ reste encore le plus souvent superieur k R^ malgr6 la
margarine ou les deux rapports deviennent au plus 6gaux.

Mode operative

Je vais indiquer rapidement comment j'isole une quantity
suffisante d'acides gras insolubles dans I'eau pour determiner en
suite leur indice de saponification.

II consiste k suivre la technique expos^e dans le premier
m^moire, avec cette difference, que je pr616ve 10 grammes de
beurre fondu et filtr^ au lieu de 5 grammes; prends un ballon
jaug6 k 300 au lieu de 150, je saponifie avec 50 cent, cubes de
liqueur de saponification au lieu de 25, et j 'ajoute 34 cent, cubes
d'eau au lieu de 17, pour amener le titre alcoolique k 56°5.

Apr^s la filtration k la temperature de 15°, je dose les acides
gras solubles dans I'alcool a 56°5 et ceux solubles dans I'eau,
sur le filtratum, exactement comme je le fais savoir dans le ler
m^moire. Ce qui reste de liquide filtr^ est introduit dans un
b^cherglass de 250 et ^vapor^ au bain-marie jusqu'S, reduction
au volume de 45 cent, cubes. La liqueur concentr^e, sur laquelle
les acides gras insolubles dans I'eau surnagent, est vers^e etant
chaude, (car la filtration est alors plus rapide) sur un petit
filtre sans plis que Ton a pr^alablement echauff^ lui-m^me, en
le remplissant d'eau bien chaude.

Apr&s avoir lav6 le b^cherglass avec de I'eau chaude, pour
entrainer le plus possible d'acides gras qui y adherent, ceux-ci
sont lavfe aussi 4 ou 5 fois sur le filtre avec de I'eau bien chaude.

On laisse alors le filtre s'^goutter 2 ou 3 minutes, puis k I'aide
d'un tube de tr^s petit diam^tre, et a extr^mite l^g^rement

xvra] Congress of Applied Chemistry 307

effil6e, on pr616ve les acides gras retenus sur le filtre, pour les
introduire dans un petit ballon ta,v6 (1).

Une nouvelle pesfie fait connattre le poids de ces acides.

On les dissout par addition de 20 cent, cubes d'alcool neutre
k 90° 95° et aprSs agitation on prend I'acidit^ en presence de
2 gouttes de solution de phtal6ine, en se servant de potasse
d^ci normale dont on cesse I'addition, d^s que se manifeste le
virage au rose trSs faible.

Supposons avoir op6r^ sur 0 gramme 453 d'acides gras et que
leur saturation a n^cessit^ 20 cent, cubes de potasse d^cime;
I'indice de saponification sera donn6 par le calcul ordinaire k

savoir:

20X0.0056 „._,,. ,. , ._ ,.

— fTTco — = 247 d mdice de sapomncation.

Le tableau suivant donne une id4e des renseignements que
peut fournir I'application des operations que j'ai cities.

II est bien certain que si Ton peut ainsi d^coavrir 5% de coco,
on comprend qu'^ fortiori, il sera ais6 d'en trouver une teneur
plus 61ev4e; le rapport R^ pourrait ^tre utilement cherch^;
du rests sa determination ainsi que Ton a pu s'en rendre compte,
peut se lier k la pratique de ma m^thode rapide d'analyse des
beurres (voir ler m^moire).

Ri R«

B.P 38 43

B.C 40 35 abr^viation

B.CM 41 38 B.P. -beurre pur

B.P 34 37 B.C. - beurre -^ 5% de coco

B.C 36 34 BCM. -beurreH-5% de coco-J-5%

BCM 38 36 margarine

(1) II est bien de chauffer un peu ce tube, pour que les acides ne s'y solidi-
fient point.

THE CHEMIST IN THE SERVICE OF THE PACKING

HOUSE

Paul Rudnick
Chicago, III.

There is perhaps no other industrial line of work embracing so
great a variety of subjects as packing house chemistry. This
work may vary from the routine analysis of occasional samples
of a few fats and fertilizer materials such as blood, bone and
tankage, often performed under contract by a commercial
chemist, to the great variety of work performed in the laboratories
of the larger packing houses by a corps of chemists and trained
helpers.

Confining the subject under discussion to the larger labora-
tories, the work may be classified in a general way into three
fairly distinctive lines, namely analytical, research, and consult-
ing work.

The analytical work embraces two principal lines; first, the
examination of purchased materials, so far as they can be pur-
chased on specifications involving chemical control, and second,
the control of the finished manufactured products, often includ-
ing the various stages of manufacture as well. Under the head
of purchased materials may be mentioned, for example, steel,
iron and cement for construction work; paper, tin plate, wood,
fiber board, jute, and burlap, for containers or wrapping purposes;
and such raw materials as edible and inedible fats and oils,
phosphate rock, and potash salts, which enter into the manu-
facture of finished goods.

Very often unusual specifications have to be made for pur-
chased materials, as for example in the case of special coverings
or special containers for meats which have to withstand the
deteriorating infiuence of tropical, humid climates.

In the control of products manufactured in the packing house,
one of the principal aims in the case of edible products is, of course,
to comply fully with the letter and spirit of the foreign and

309

310 Original Communications: Eighth International [vol.

domestic laws, including the Meat Inspection Regulations, the
Foods and Drugs Act, and the laws of the various individual
states. It is also important, however, that the product shall
suit the consumer, who often enough seems to know better what
he doesn't want, than exactly what he does want. Local
customs, Ukes, dislikes and often prejudices on the part of the
consumer and retailer must be met, and as in other selhng transac-
tions, this is often possible only by an analysis or examination of
a sample of the product which suits the consumer.

In the long list of work on edible packing house products may
be included the control of the fresh, cured, smoked, and canned
meats, sausages, edible fatsi, and the processes by which they are
produced. To this may also be added the animal products,
used chiefly for medicinal purposes, such as pepsin, pancreatin,
desiccated glands and their active principles.

Aside from the routine analytical work there are often special
analytical problems presented in the control of manufacturing
processes. In the manufacture, for instance, of pemmican for
polar expeditions it is absolutely necessary to have not only a
well-balanced ration of protein, fat and carbohydrates, but the
percentage of moisture in the meat, the sweetness of the meat
and fat, the amount of salt, spices, sweetening, etc., must be
rigidly controlled. Again, in an emergency methods can and
must be changed or adapted to meet the situation. Some years
ago, for example, it became necessary to determine the absence of
boron compounds in not less than 500 samples of cured meat
daily. To incinerate and extract such a number of samples daily
was impossible with the equipment and help at hand. A number
of comparative analyses confirmed the idea that the qualitative
test for boric acid could be made on the brine in which the meats
had been cured by simple treatment with hydrochloric acid, and
direct appHcation of the Goske method, the results being just as
satisfactory as if the determinations had been made on the ash of
the meat. In this manner it was easily possible for one man to
carry on the work at the rate of 500 samples daily.

In the case of soaps, glues, fertihzers, inedible fats and oils,
and other inedible products such partial or complete analyses
are made as will assure the selling department that the product

xviii] Congress of Applied Chemistry 311

will fully meet with the requirements of the purchaser and his
specifications, or in the case there are no such requirements or
specifications that the products will prove acceptable for the
purposes for which they are intended. In a very large part of
the work the highest accuracy must be sacrificed to a certain
extent to the necessary speed. For instance, it is far more
important to avoid per diam penalties or so-called demurrage
charges on cars held on the track, to secure cash discounts on
materials bought subject to analysis, to avoid costly interruptions
in manufacturing processes, the adjustment of which depends
on analyses made on the finished product, or to furnish a tele-
graphic quotation on the day of the inquiry, than to employ
unnecessarily refined methods of analysis.

Where so much analytical work must be done, economy of
materials and of operation becomes a very important matter.
It is necessary, for instance, to arrange the apparatus, for determi-
nations which have to be made in great numbers, in the most
compact, simple and convenient form possible, so that the opera-
tor will not have to waste time in carrying out the different steps
of his work. The expense of reagents can often be materially
reduced. For example, the potassium sulphate for nitrogen
determinations can be bought from the wholesale druggist in the
powdered form in which it is used in medicinal preparations,
much more cheaply than the so-called chemically pure article of
the chemical supply houses. The only requisite in this case is
its freedom from nitrogen, and this must be determined in any
event by blank determinations. Again, ammonium nitrate,
which is used so largely in phosphoric acid determinations, can
be purchased at a relatively low price, if the specifications
regarding its purity be limited to absence of phosphates.

It may not be amiss to refer to a mistaken notion which seems
to have grown up regarding the value of such analytical work, a
notion which has crept even into undergraduate life, namely that
analytical work is something to be shunned because of the lack
of opportunity for advancement. Nothing coxild be further
from the truth. There is no more valuable training for the indus-
trial chemist and chemical engineer than that which he can
obtain in such work as has been described above. If he will

312 Original Communications: Eighth International [vol.

stop to realize what an important bearing even the simplest
routine operation may have on manufacturing processes and how
often large money values are involved and depend on the faith-
fulness with which such a simple routine determination is carried
out, then his work will take on a new meaning, and actual experi-
ence shows that he will make a better man in every way, other
things being equal, than the one who lacks such experience and
training. Advancement simply depends upon whether or not
he is content to remain a routine analyst.

Little can be said regarding the research work which falls to
the lot of the packing house chemist. Like all industrial research
work it is, from its very nature and purpose, confidential in
character, although more frequently than ever before, certain
phases of it may be of sufiScient scientific interest to bear publica-
tion without destroying the value of the general proposition at
hand. If this is so, it will doubtless be due to the fact that the
methods employed are becoming more and more scientific and
less empirical than they have been.

It is out of the question to indicate even in the briefest way
the variety of subjects considered in the consulting work required
of the packing house chemist, except perhaps, to say that the
executive heads, general and department superintendents, engi-
neers, architects, attorneys and many others daily refer questions
to the chemist for answer. Many of these subjects are of the
greatest interest, sometimes involving careful and extended
experimental work, at other times a carefully planned series of
analytical determinations or of microscopical or bacteriological
work. Occasionally, however, emergencies arise where decisions
must be made on the spur of the moment without time for a
search of the literature or for actual experiments, otherwise the
time for decision would be past.

In conclusion, I cannot emphasize too strongly a fact quite
commonly overlooked by the chemist, namely that his work is of
no value to the business man unless results, conclusions, judg-
ments, or opinions be reported in simple, clear and concise lan-
guage, avoiding as far as possible the use of technical, involved,
or indefinite expressions, or conditional statements.

AN INVESTIGATION ON THE MANUFACTURE OF TEA

By S. Sawamura
College of Agriculture, Imperial University, Tokyo

I. Effect of Steaming on the Activity of the Enzyms of
Tea Leaves

In green tea leaves there are present abundant oxydising
enzym, wherefore Mann in India holds an opinion that oxydising
enzym is one of the factors which determine the quality of tea.
In the manufacture of green tea, however, oxydising enzym
of tea leaves is killed by steaming, because when it is active
the green color of tea leaves can no more be retained. The
author^ foimd in another investigation that the formation of
some aroma of manufactured tea, which takes place usually
during the rolling of tea leaves, is due to the action of a certain
enzym on a certain compound of tea leaf. Hence if steaming
kills all the enzyms of tea leaves the production of aroma may be
more or less hindered. .

In 1909 I tried to know whether all the enzyms of tea leaves
lose activity by steaming in the usual manner. In these trials
green leaves were steamed in the usual manner, respectively
for 30 seconds, 50 seconds and one minute, and the steamed
leaves as well as the unsteamed were crushed and extracted
with 40% alcohol. The extracts are precipitated with ether-
alcohol and filtered. The precipitates were washed with alcohol
and again dissolved in water. The solution gave no reaction
with FejCle, proving the absence of tannin. Oxydising enzyms
were tested with guayak tincture, and guayacol and H2O2, by
which the solution obtained from unsteamed leaves showed the
characteristic reaction, while the steamed did not. Steaming for
30 seconds killed oxydising enzyms completely. In another trial
tea leaves steamed for 20 seconds were tested for the presence
of oxydases, and a faint reaction was observed. From these

' Bulletin of Agric. Exp. station, No. 1.

313

314 Original Communications: Eighth International [vol.

facts we know that the oxydising enzyms of tea leaves lose activity
when they are steamed only for 30 seconds.

I tried then to see whether the enzyms other than oxydase
lose activity by steaming for a short time. Preliminarily I
detected diastase in tea leaf by the following manner. Green
tea leaves were crushed in a mortar and extracted with 40%
alcohol. To the extract ether-alcohol was added, and the pre-
cipitate thereby formed was washed and again dissolved in water.
In this solution tannin was removed by hide powder and putre-
faction was prevented by the addition of thymol. It was filtered,
and the filtrate which gave no reaction with Fe2Cl6 and did not
reduce Fehling's solution, some boiled starch and thymol were
put in. The solution after having been kept at 40° C. for 4 days,
reduced Fehling's solution considerably. We confirmed by this
trial that diastase of tea leaves can be detected in this manner.

The tea leaves steamed for 30 seconds, in which oxydase was
completely killed, reduced also Fehhng's solution when treated
in the same manner. Hence we know that oxydase is much more
sensible than other enzyms such as diastase and it is highly
probable that some enzymatic actions take place in the first
stage of rolling tea leaves, and the production of some fine aroma
is due to them. In practice, therefore, steaming of tea leaves
must be so regulated as to kill only oxydising enzyms but not
other enzyms.

II. Effect of Rolling on the Solubility of Tea

Whether the object of rolling tea leaves in the manufacture
of green tea is to give tea a fine shape or to press out the juice
in order to accelerate the desiccation of the leaves, or to break
the cells in order to increase solubility is, as far as I know, not
yet decided. According to the investigation of Dr. Kozai'
the solubihty of green tea was little increased by the manufacture,
but Rombe and Roman's experiment^ showed on the contrary
the decrease of soluble tannin and thein.

To settle this question I made an experiment in 1905, in which

» Bulletin of College of Agriculture and Dendrology No. 7.
' Kdnig. Chemie der Nahrungs und genussmittel B. II.

XVIIl]

Congress of Applied Chemistry

315

fresh tea leaves, picked at a sheltered tea garden, were divided
into three parts, and one of them was steamed and dried without
rolling which served as control; the second part was prepared
into green tea (Gyokuro), and the third part into Tencha, which
is usually prepared without rolling the leaves. The infusion of
these three kinds of tea was found to be as follows: —

Control

Tencha

Gyokuro

Color

Flavor

Taste

light
weak
faint

deeper

Btronger

good

deepest

strongest

best

The reaction of the infusion with FejCle was not the same in
three kinds; that of Gyokuro produced deep black color, while
control and Tencha a very faint black color. The solubility of
tea was determined as follows: — 400 cc. of boiling water were
poured on 10 gr. of the powdered sample which had been kept
at 100° C. for an hoxir. It was filtered after leaving it to stand
for 5 minutes and washed on filter with 100 cc. of boihng water,
and soluble matters were estimated in it.
The composition of the control tea was as follows : —

In 100 pts. of air dry substance

water

6.215

In 100 pts. of dry substance

Crude protein

41.984

Albuminoids

28.252

Ethereal extract

9.042

Crude fiber

12.012

Nitrogen free extract

14.101

Thein

3.529

Tannin

15.968

Crude ash

6.883

Total nitrogen

6.717

Albuminous nitrogen

4.520

Thein nitrogen

0.934

Amide nitrogen

1.263

316 Original Communications: Eighth International [vol.

The soluble constituents of the three samples were as follows: —
IlSf 100 PTS. OP DRY MATTERS

Control

Tencha

Gyokuro

Dry matter

Tn.TiTiiTi

34.057
7.083
3.124
5.249

34.130
6.939
2.996
5.373

33.862
6.477

Thein

3.088

Ash

5.197

According to these results Gyokuro, which was prepared by
rolling the leaves, showed no greater solubility than the other
two. Soluble tannin decreased in Gyokuro probably in conse-
quence of oxydation during the rolling.

In the other experiment I determined solubiUty of three
samples in a different manner. 10 gr. of whole, not powdered
sample were put in a beaker, and after keeping it at 100° C. for
an hour 200 cc. of boiling water were poured on and filtered
through glass wool after leaving it to stand 5 minutes. In the
filtrate dry substance, crude protein, tannin, thein and ash were
estimated. They were as follows: —

Control

Tencha

Gyokuro

In 100 part of air

dry

substance

Water

8.375

In 100 pts. of

dry

matter there

were soluble

Dry matter. .

16.076

1.885

Tannin

0.659

Thein

1.975

Ash

3.405

7.953

21.190
2.141
1.312
2.243
4.411

7.638

29.233
2.313
5.492
2.804
4.385

xvinl

Congress of Applied Chemistry

317

In 100 pts. of each
constituent there
were soluble

Dry matter ....

Nitrogen

Tannin

Thein

Ash

Control

17.645
28.068
4.127
65.965
49.760

Tcncha

23.021
31.869
8.216
63.559
64.083

Gyokuro

31.656
34.427
34.374
79.453
63.708

The increase of solubility compared with the control was found
to be as follows: —

Gyokuro

Dry matter
Nitrogen. .
Tannin ....

Thein

Ash

14.111
6.359
30.247
23.488
13.958

We see that, when the whole, not powdered samples were used,
there were greater increase of solubility in the rolled leaves.
Hence we may conclude, that the rolling of tea leaves has the
effect of increasing easily soluble matter by crushing the cells
and pressing out the juice and making it dry on the surface of the
leaves.

Second experiment on the same subject was carried on in 1906
with tea leaves picked in unsheltered tea garden. The leaves
were divided into two parts, and one part was dried after steam-
ing and served as control, and the other part prepared into green
tea. The infusion of the two samples was foxmd to be as fol-
lows:—

Control

Green Tea

Color

For lighter
Nearly null
Faint

Common

Flavor

Good

Taste

Good

318 Original Communications: Eighth International [vol.

The composition of the original leaves was found to be as
follows : —

In 100 pts. of air dry substance

water
In 100 parts of dry substance

Crude protein

Ethereal extract

Tannin

Thein

Ash

Soluble matter

Total nitrogen

Thein nitrogen

6.008

33.209

25.656

18.889

3.266

5.719

44.525

5.313

0.864

The solubility which was determined in whole, not powdered
samples, was foimd to be as follows : —

Control

Green tea

Drv matter

9.879
0.969
4.883
1.995
1.383

26 692

1.410

TaTlTllTl

12 802

Thein

2.136

Ash

3 077

Control

Green tea

Increase in
Green tea

In 100 pts. of each constitu-
ent there were soluble

Dry matter

Nitrogen

Tannin

Thein

Ash

22.119
18.227
25.850
61.074
24.186

69.948
26.531
67.778
65.403
53.811

37.829
8.304

41.928
4.329

29.625

XVIIl]

Congress of Applied Chemistry

319

The result of this trial agreed with that of the former one,
showing the increase of easy solubility in the rolled leaves.
Hence we may conclude, that the chief effect of rolling tea leaves
is the increase of easy solubility of the constituents. The desic-
cation of the leaves will also be accelerated by rolling by press-
ing out the juice from the interior of the cells. From these facts
we are justified in testing tea-infusion to take whole, not powdered
sample, and to infuse it only for a few minutes. Total solubility
as was determined in the usual method is not of much use for
practical purpose.

III. The Effect of Fihinq on the Chemical Composition

OF Tea

Green tea as well as black tea are usually refired some days
later after the manufacture. By refiring the flavor is much
improved, but the infusion becomes usually darker in color.
In 1908 and 1909 I made some investigation on the effect of re-
firing on the quality and composition of tea. I kept respectively
green tea and black tea at various temperatures for one hour and
then analyzed. Tannin was estimated by Lowenthal's method
and thein by Mulder's method. Solubility was determined by
infusing 2 gr. of whole tea leaves in 400 cc. of distilled water for
2 hours, and after 100 cc. of water had been added it was filtered.
The temperature used for firing, the color and flavor of the in-
fusion and the color of the infused leaves were found to be as
follows : —

1908

1. GREEN TEA

Color of the

No.

Temperature

Color

Flavor

infused leaves

Control

Little lighter than

Weaker than

(not fired)

No. 3

No. 2

Greenish yel-

61°C

Nearly same as No. 3

Best

low

82°C

Best

Little too strong

101°C

Rather red

Little reddish

123°C

Reddish than No. 4

Bad smell

Reddish

140°C

More reddish No. 5

160°C

Reddish

Blackish brown

320 Original Communications: Eighth International [vol.

2. BLACK TEA

Color of the

No.

Temperature

Color

Flavor

infused leaves

Control
(not fired)

Lighter than No. 2

Weaker than
No. 2

62°C

Lighter than No. 5

Weaker than
No. 3

Brown

81°C

Lighter than No. 1

Best

101°C

Lighter than No. 3

5
6

iig-c

141°C

Most reddish
Lighter than No. 4

Bad smell

Blackish
brown

166°C

Lighter than No. 6

1909
1. GREEN TEA

Color of

Aroma of

Taste of

Color of the

Mo.

Temperature

infusion

infused leaves

Control
(not fired)

Faint

Weak

60°C

Best

Weak

Astringent

70°C

Lighter than
No. 4

Best

Good

Usual

80°C

Lighter than
No. 2

Good

Best

90°C

Reddish

Bad

Bitter

100°C

Worst

Most bitter

Little burnt

2. BLACK TEA

Control
(not fired)

Not clear

Faint

Weak

2
3

60°C
70°C

Light

Best

Good
Best

Usual

80°C

Best

Weaker than

No. 3

90°C

Worse than

No. 4

Bad

100°C

Blackish

xvm]

Congress of Applied Chemistry

321

The chief constituents of the tea \frere found to be as fol-
lows:—

1908

1. GREEN TEA

No.

Temperature

In 100
pts. of
air dry
subst.

In 100 pts. of dry substance

Water

Tannin

Thein

Solu-
bility

Soluble
tannin

Soluble
thein

1
2

Control

(not fired)

61°C

82°C

101°C

123°C

140°C

160°C

5.228

4.633
3.158
1.383
2.045
2.453
3.005

15.690

3.210

37.458

36.786
35.417
35.553
37.364
35.162
29.898

11.724

11.906
11.785
11.688
11.096
10.400
6.470

2.506
2 600

2.601

4
5

14.602

3.101

2.444
2.455

2.287

13.248

3.098

2.317

2. BLACK TEA

Control
(not fired)

62°C

81°C
101°C
119°C
141°C
156°C

4.445

3.985
3.703
2.293
4.875
2.230
1.460

8.575

3.075

27.393

27.027
27.415
27.020
26.281
24.957
22.757

4.045

4.151
4.390
4.450
3.682
2.594
1.961

2.518
2.676

2.290

7.247

3.130

2.705
2.477

2.443

5.797

3.135

2.500

322 Original Communications: Eighth International [vol.

1909
1. GREEN TEA

No.

Temperature

In 100
pta. of
air dry
Bubst.

In 100 pts. of dry substance

Water

Tannin

Thein

Solu-
bility

Soluble
tanniii

Soluble
thein

2
3

Control
(not fired)
60°C
70°C
80°C
90°C
100°C

5.157

4.512
3.903
3.168
2.105
1.844

15.857
15.844

3.077
3.134

37.557

37.252
37.463
36.455
35.569
24.666

11.621

11.883
11.936
11.711
11.236
11.451

2.191

2.425
2.483

4
5

15.418

3.209

2.536
2.442

14.586

3.061

2.487

2. BLACK TEA

Control

(not fired)

60°C

70°C

80°C

go-c

100°C

6.320

4.541
3.574
3.218
2.271
2.049

8.484
8.632

3.165
3.163

27.556

27.137
27.378
27.098
26.727
26.475

3.917

3.844
3.955
3.979
3.479
3.283

7.402

3.147

7.093

3.046

2.348

2.456
2.403
2.407
2.221
2.200

From these results we may conclude, that green tea is improved
in quality by being fired at 70° C. for one hour, and temperature
higher than 70° C. spoils both the flavor and color. The optimum
temperature for firing black tea hes little higher than for green
tea; viz. 80° C , and Uke green tea higher temperature spoils the
flavor and color. By reflring tannin and thein decrease more or
less, probably the former being due to oxydation and the latter
to volatilization. Solubility increases little when tea is not
strongly heated, but when temperature is high total soluble
substance and tannin decrease remarkably. Therefore in firing
tea temperature must of course be properly applied. If it is
too high, the quality of tea is much deteriorated.

WHEAT FLOUR. A MONOGRAPH

Harry Snyder, B. S.
Minneapolis, Minn.

In considering the chemical composition of a flour, its moisture
content and the basis upon which the results are reported, are
matters of first importance. Flour freshly milled may contain
from 12.50 to 13,50 per cent of moisture. When stored under
the best conditions its moisture content appreciably decreases.
Often the chemist receives for analysis a small, over-dried sam-
ple sent in paper envelope. This sample may contain less than
8% moisture. Comparison of such a sample with one freshly
milled is inconsistent unless the necessary corrections for dif-
ferences in moisture content be made. This difference in moist-
ure content, unless corrected, disturbs comparative analytical
results and is the occasion of much unfavorable comment as to
the value of a chemical analysis of flour.

The extent to which a variation of 6% in moisture may affect
results is large. A freshly milled flour with 13.50% moisture
and .40% ash, would when dried to 7|% moisture show nearly
.43% (.427) ash:

The maximum standard of 13.50 per cent of moisture adopted
by the U. S. Department of Agriculture is reasonable when the
wide range in moisture content of wheat is considered. As an
illustration of this range, 278 cars of wheat tested for moisture
by myself during the present year showed 48 cars with less than
13.5 %moisture and 230 cars with more than 13.5% moisture.

The question of the moisture content of flour is a disturbing
factor not only in chemical analysis but also in the matter of
weight of a package. Any loss of moisture causes a propor-
tional loss of weight. Since the Government has established
the maximum moisture content of flour at 13|%, that neces-
sarily establishes the minimum dry matter at 86|% and in turn
determines the approximate tolerance allowable for shrinkage in

323

324 Original Communications: Eighth International [vol.

weight. To illustrate: if a 98-lb. package of flour after storage
and handling weighs 97 lbs. and is carefully sampled and its
moisture content found to be 11|%, it means that the 97 lbs. on
an 11|% moisture basis contains 85.84 lbs of dry matter. If
the flour had been packed with 13.50% the 98-lb. package would
have contained 84.77 lbs. of dry matter. As it is, it contains
about one pound of dry matter or dry flour more than is called
for on the minimum dry matter basis and hence cannot be con-
sidered as short in weight. On the basis of dry matter it is not
short in weight and when packed it did not contain the maxi-
mum moisture content allowed by the Government standard.
Hence in the consideration of both weight and chemical compo-
sition of flour the moisture content is a matter of first import-
ance. All comparisons of composition should be made on a uni-
form moisture basis. The extent to which a flour sample may
dry cannot be anticipated and the moisture removed by drjdng
must in turn be added in bread-making. Thus the consumer is
in no way defrauded by the drying of flour provided it is packed
full weight and without excessive moisture, that is above 131%.
Indeed more water than that would endanger the keeping quali-
ties of the flour and entail loss on the part of the manufacturer.
Flour with either an excessively low or high moisture content is
not normal flour.

Over-dryness of flour may affect the analytical results by trans-
location of soluble ingredients. In the case of a sample of flour
freshly milled with .40% ash, about one-fourth is mechanically
combined^or chemically united with the gluten, one-fourth with
the starch and one-half is capable of being dissolved in distilled
water. When a flour dries an uneven distribution of the solu-
ble constituents may occur, depending entirely upon governing
conditions. Thus if a sample of flour be drawn from only one
part of a large flour package it may show an abnormal ash
content.

XVIIl]

Congress of Applied Chemistry

325

Example oj Translocation of Flour Ash.

Ash Content
of Flour

Moisture
Content*

Fresh Flour 140-lb. packages
Sample from exterior of pack-
age after storage 3 mos. in
flour store room

.37+

.41
.37

12.92 .37+
10.84 .40+

Sample from Center of pkg. .

11.58 .36+

Drying of a flour accompanied by translocation, and drawing
of the sample from a portion only of the package may affect
the ash results to the extent of .06 of a per cent. Not infrequently
do the results of two laboratories reporting on the same sample
of flour show as large and even a larger difference than .06%.

In the determination of the ash of flour other errors that
may occur result from : too high a temperature during combustion,
incomplete combustion, fusion and occlusion of particles of
carbon and failure to make the necessary distinction between
crude ash and pure ash. The temperature during combustion
should be appreciably below 675°C. and the combustion should
be continued until a light grey granular ash is secured, reasonable
constant in weight. The ash should not be fused and should be
corrected for carbon and combined carbon dioxide. The ash
from a refined flour can be obtained quite free from carbon and
combined carbon dioxide, so that there is no appreciable differ-
ence between the crude ash and the pure ash. But, if the com-
bustion is not made with care, the difference is large.

The main loss caused by high temperatures for combustion
is sulphur. This, however, affects the ash percentage less than
.01 per cent. There appears to be no loss of phosphorus during
the combustion process at low temperature as there is sufficient
alkaline matter to form non-volatile pyro phosphates. In fact
I have been uuable to obtain any difference in the phosphorus

•Ash on basis of uniform moisture content.

326 Original Communications: Eighth International [vol.

content of a flour from the analysis of the ash, and from the
analysis of the residue of the calorimeter where the flour is burned
in such a way as to preclude any possible loss of phosphorus
compounds.

No official method has yet been adopted by the Association
of Official Agricultural Chemists for the determination of the
ash of cereals. In the work on ffours and cereals by the U. S.
Department of Agriculture a just distinction has been made
between crude ash and pure ash along the lines laid down by
European Chemists.

Methods have been proposed for the complete analysis of the
inorganic constituents of plants in which calcium acetate is used
to prevent volatihzation of sulphur, and then corrections are
made for the lime and carbon dioxide introduced by its use. The
application to flour of such a method for obtaining ash is not
feasible as corrections for carbon dioxide &c. must be made on
each individual sample tested. This in turn calls for the com-
bustion of 100 grams of flour so as to get enough crude ash to
determine the quantities of impurities to be deducted. Such a
method would be impracticable in flour mill work. No common
factor could be assumed for correction as a more or less richly
carbonated ash of a variable degree of purity is obtained. The
acetate introduces one-fourth as much mineral matter as is
naturally present in the flour and its use may occasion the intro-
duction of a larger error than* it is intended to correct: viz.
volatilization of sulphur.

When made under uniform conditions and by one person, the
ash results indicate the mechanical uniformity of a flour to a
high degree and are of value in flour mill control work but
alone — the ash results are incapable of determining the bread-
making value of a flour.

When the ash results of two chemists working on the same
sample of flour are compared wide differences may be observed
because of variation in moisture content, occlusion, transloca-
tion, incomplete combustion and failure to make the necessary
distinction between crude and pure ash. These variations are
often so large, and unnecessarily so, as to cast much discredit on

* Through formation of carbonates.

xvni] Congress of Applied Chemistry 327

chemical tests and their application to determining the value of
flour.

Another source of confusion in the interpretation of a flour
analysis is the occasional use of the factor 5.7 for converting
total nitrogen into protein, instead of 6.25 and then no mention
being made of the factor used. In technical scientific investiga-
tion the use of special factors for protein determination is neces-
sary and unquestionable, but to use a special factor for wheat
and a general factor (6.25) for all other foods is inconsistent.
From a nutritive point of view the 6.25 factor for wheat is more
correct than 5.70 because the wheat proteins are concentrated
in nitrogen containing 17.50% against the general average of
16.25. It is the nitrogenous part of the molecule which gives
the unique food value to the proteins, and in the wheat proteins
the consumer gets more of this material. In fact the 5.7 factor
assigns too low a nutritive value to wheat. Wheat proteids are
too concentrated in distinctive nitrogenous material to be as-
signed so low a percentage value — when compared with other
foods where the proteins are of lower nitrogen content. It is
the quality of the protein that determines its nutritive value
as well as the amount, and in wheat the proteins are of strong
nitrogenous character.

The general method for the determination of crude fiber is not
satisfactory for determining the fiber content of flour, as the
action of the acid and alkali solutions for the removal of non-fiber
materials is not complete. By extracting the flour with
70% alcohol, after extraction with ether, better results are
obtained as the gliadin is removed, foaming is prevented and
the material is in better mechanical condition for extraction with
acid and alkali solutions. This extraction with alcohol is bene-
ficial in determining the fiber of wheat products.

In conclusion, it may be said that the chemist can do most in
the way of flour investigations by making a study of bread-making
processes and the factors which control them. Flour making is
distinctly a mechanical process and the whole tendency of modern
flour manufacture is in the direction of producing cleaner flom-.

Since wheat flour takes such an important part in the dietary
and because it supplies such a large amoimt of nutrients at com-

328 Original Communications: Eighth International [vol.

paratively low cost, it is consistent that the efforts of the chemist
be directed toward encouraging the farmer to raise the best of
bread wheats in the most approved ways through scientific
agriculture, and that a broader knowledge be secured and gen-
erally disseminated concerning the principles of bread-making
and the nutritive value of bread and foods in general. In this
work the food chemist must necessarily take the leading part.
If indifferent, he is not doing his duty.

ON SOME DRIED MILKS AND PATENT FOODS

By a. W. Stewaht, D.Sc.
West Hampstead, London, N.W., England

Whether dried milks, infants' foods and other milk food prepa-
rations are becoming more popular or that mothers find it more
convenient to bring up their children on these products, the
fact remains that there is a steady increase in the number of
these preparations. Although some are of a wholesome nature
there are on the market nevertheless a large number whose chief
ingredient is starch, contained in such a proportion as to be
totally unsuitable for infants. Artificial products cannot effec-
tually replace what nature has supphed. The characters of
human milk are such that its imitation seems almost irrealis-
able. It is true that by diluting cows' milk with water and
adding lactose and cream, an article can be produced much
resembling human milk, but it has not those properties that
render mothers' milk the ideal food for children. Sommerville
pointed out the value of dried milk for infant feeding on account
of its relative sterility and the absence of a dense clot in the
infant's stomach. When one studies the results of the analyses
of the infants' foods on the market, the unsuitabihty of the
majority of them is greatly in evidence. The AustraUan Food
Standards Committee 1906, recommended that "Infants' food
shall contain no woody fibre, no preservative substance and no
mineral substance insoluble in water; and unless described or
sold specifically as a food suitable only for infants over the age
of seven months shall, when prepared as directed by an accom-
panying label, contain no starch and shall contain the essential
ingredients of and conform approximately in composition to
normal mothers' milk." Apart from the high starch percentage
of a number of brands, there are not many which form even in
10% solutions anything approaching a homogeneous solution.
In several cases this was not possible, the result being a thick
12 329

330 Original Communications: Eighth International [vol.

pasty mass caused by the swelling and cohesion of the starch.
The small % of fat is an outstanding feature of inferior brands;
they are deficient in the chief body fuel namely fat. Abun-
dance of fat should be the main characteristic of the diet of
infancy just as abundance of carbohydrates is the chief feature
of the adult and laborious life. The frequent connection between
rickets and deficiency in fat is an undeniable clinical fact.
Again, the desiccating process destroys the enzjones always pres-
ent in raw milk and to which its anti-scorbutic properties are
generally ascribed.

Preparation

Until lately the process of separating the soHd and liquid
constituents of milk was too costly to render the manufacture
of "dried milk" a profitable industry. The machine used in the
"Just-Hatmaker" process appears to give the most satisfactory
results; it consists of two large metallic drums 28 inches in
diameter and 6 feet long, mounted horizontally in a framework
with a space of about fth inch between them. High-pressure
steam, admitted to the drums through axial pipes, raises their
surfaces to a temperature of 220° F. The milk is allowed to
flow in thin streams over the revolving drums, the heat of which
quickly evaporates the water. A coating of solid matter gradu-
ally forms which is scraped off by a knife and falls into a recep-
tacle. The milk is not boiled, though completely sterilised by
the heat. A slight decomposition of the proteins and fat prob-
ably takes place.

Classification

1. Dried milks and milk products; they consist of milk
evaporated and dried, with or without the addition of lactose
and fat. They are free from starch. The results are given in
table I.

2. Farinaceous and malted foods; in this are included foods
containing either malt or starch or both. In many cases the
diastase was not active. The results are given in table II.

3. Miscellaneous products, oats, barley, etc., for growing
children. The results will be found in table III.

xviii] Congress of Applied Chemistry 331

Methods of Analysis

Water: This was estimated by drying 10 grams of the sample
at 100° C. for 4-6 hours.

Ash: The residue was incinerated and weighed in the usual
manner.

Total Pid: Where it was desirable to estimate the phos-
phates, a titration with N-10 uranium nitrate using K4Fe (CN)6
as indicator was made.

Total proteins: This was estimated by the usual Kjeldahl
process. The factor N x 6 . 38 was used unless otherwise stated.
A verification of this factor was effected by D. Richmond (Ana-
lyst, 1908, p. 179). When the nitrogen is present as casein or
albumen the factor 6.38 should be used but when its origin is
uncertain 6 . 25 is recommended as a general factor.

Soluble albumen: 10 cc. of a 10% solution are digested with
20 cc. of saturated MgS04 solution and crystals of magnesium
sulphate are added until they no longer dissolve. An excess
does not matter, provided the MgS04 is free from Na2S04. The
solution is put aside till the next morning, when it is filtered and
washed with a little saturated MgS04 solution. The filtrate is
then treated with Almen's reagent (4 gram tannic acid, 190 cc.
50% alcohol, 8 cc. 25% acetic acid) and the precipitate allowed
to settle till the next day. It is then filtered, paper and con-
tents transferred to a Kjeldahl flask and the estimation of the
proteins carried out in the usual way.
Lactose and Carbohydrates. These were obtained by difference.

Fat: We utihsed 3 methods: (1) Leffmann and Beam, centri-
fugalisation of fat (2) Adams and Soxhlet process, the latter in
certain cases where the Adams process could not be applied
owing to the product not forming a solution with water (3)
Werner-Schmidt. The last named is the only reUable method
combining accuracy with rapidity. The Adams and Soxhlet
extraction were often found to be unrehable. This is in agree-
ment with McLellan {Analyst, 1908, p. 353) ; he found that the
incompleteness of the extraction of the fat is due to the coating
of the fat globules in the milk during the process of evaporation
with an impermeable substance which prevents the solvent
from penetrating. He found that it was possible to completely

332 Original Communications: Eighth International [vol.

extract the fat from dry milk in a soxhlet apparatus if the sample
was soaked overnight, then extracted for 8 hours and allowed
to soak again overnight and finally extracted for 1 to 2 hours.
That the results obtained by the Adams process are too low is
further borne out by Siegfeld {Molkerei Zeitung, 1909, N. 25),
Grunhut {Zeit f. Anal. Chem., 1911, p. 649), and Anton Burr
(Milchwirtschaftl. ZentroM., 7, p. 118). The latter found the
Werner-Schmidt and Rose-Gottlieb processes to give good
results, the last named requiring special conditions to be com-
plied with according to D. Richmond {Analyst, 1908, p. 389).

Physiological Fuel Value: The results are expressed in large
calories per 100 grams of the sample. The factors used were
fat 9, protein 4, and carbohydrate 4, these being the physiological
fuel values of food constituents.

Nutritive Ratio: is the ratio of proteid to carbohydrate and
fat, i.e., the ratio of nitrogenous to non-nitrogenous nutrients
compared on the basis of fuel values. It is deduced from the
formula (Sherman, Chemistry of food and nutrition 1911): —

carbohydrates-f-2| fat
proteins

No preservatives (formaldehyde, boric acid) were found in
any of the samples analysed.

XVIIl]

Congress of Applied Chemistry

333

■339

«3

Nutri-
tive
Ratio

*-4

CO t^

■ -^ eo ... • • CO

•3 §

CDCOCOrHCOCSOC^OOOlCt^^OOOO'^GONOO
OpCOiO(NTpSiCO>CCO>CO'^C<i'^000500
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N(NTtlOCOOO>'-i
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CO M ^ 1— * t^ t^ CO
N M t^ OJ in 00 (N

OIN(M(N'4lC0i-H(N

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iH»nOTI>5D00N»i5COt^CO'-tcOCOCOCOO5

OOI-^USOtIIOOIO

■"i-ioooooO"-<.-it~

O(M00C0t^>-iO«DO'l*00tO'*Ot0rti00c2O
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CO C^ W CO T-t i-t

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t^ »0 00 O

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334

Original Communications:

Eighth International [vol.

„, o

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XVIIl]

Congress of Applied Chemistry

335

Nutri-
tive
Ratio

1^ 00

r-
<— 1

1-
1—1

I— 1

1—1

1-H

wH i-H

TO ^ M OS

OO*-l»H»0^»-l-^t^OtD

CO t» U3 CO

6 00 -^ •*

oor-toooh-oOTHtot~oo5
eocor)<MMeoi*coco-<tieo

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CO CO CO CO

y-^ ^-v ^-x ^^

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,-,,-^.^,-.0.-<<N^r(<

to 00 ai

1-t IN N

/^tOt^00OST-Hi-(i-IC0»H

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W lO (N ■*

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la 00 00 tc

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CO ^ U5 1-1

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tOOTt<U5005r-ICOrHOtC

1> 00 ■* N

t^ IC to »o

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<3>00 -OINMt-lOOINr-l^

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lO S N S

S^?3°5!t^3SM05l3-

f- 1-1 t~ t^

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>nt>.Hi*0-*'003(NTi<c(:

lO 1-1 OJ CO

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336 Original Communications: Eighth International [vol.

o3 .3

-^ t* CD Tj<

000(NT-HCa'^0(NOSC<IOSOSC^
c01>-OtHc0I>CDcD05C0iCu0cD
WCOTdHCOCOCOCOCOCOCOCOCOCO

OCDCCTt<a'OiCQGOW3i-lT-l'«^i-(

1-^»oooo(^^c'^^cococo1-l^-Tt^
05 o o i-l

OOCOOOCDIOOOOSCDCONOSCD
r-llvOS ICi-Ht-HICtHt-Ii-I,-!

CO»OOOOSCDOcDOSi-lNcDOOCD
OOSOST-llOTHl01>05t>CDTt(rH

MOOiOOOOO

CO Ti4 lO CO l> CO

S fe o
3 § J .^ ^ .a as

m -g p g

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•3

xviii] Congress of Applied Chemistry 337

The question of starch in infants' foods

A short perusal of the results tabulated will show that nearly
50% of the samples analysed contained starch. Nearly all of
these milk food preparations are intended for infants' food, so
that this question is of capital importance in the bringing up of
children from birth. This subject was dealt with in an interest-
ing paper by Cautley. — (Lancet, Nov. 6, 1909), amongst whose
conclusions are : —

(I) A diastasic ferment is secreted by the salivary glands
and pancreas of new-born infants; the salivary secretion how-
ever is scanty in young infants and rarely appreciable before
the age of two months.

(II) Barley water contains about 2% of starch; mixtures
containing this percentgae of starch are not injurious but may
be beneficial, for the growth of lactic acid bacilli and the forma-
tion of lactic acid are thereby encouraged. These organisms
are of undoubted advantage in the prevention of the growth of
proteolytic bacteria.

(III) The evil effects of starch in early life are due to (a)
excess, (b) its admistration in the form of a more or less in-
soluble emulsion instead of as soluble starch (c) the substitution
of starch for the necessary protein, fat and salts.

Under the title "Patent Foods" an interesting pamphlet has
been written by R. Hutchison. The defects of artificial foods
may be summed up as follows : —

(I) They are often recommended when there is marked loss
of appetite; they do not promote it.

(II) It is often contended for these products that they are
more easily digested than natural foods and many of them exist
because they are pre-digested. The necessity for peptonizing
foods is greatly exaggerated. In pathological chemistry pepsin
is almost never absent from the gastric juice xmless HCL is
also absent. If HCL can be found in the stomach pepsin is
sure to be there too; there is therefore little necessity for pre-
digtisted foods.

(III) The claim often put forward for artificial foods that they
enable you to enrich the diet in certain constituents is largely
fallacious.

338 Original Communications: Eighth International [vol,

(IV) None of them is worth the money asked for it; some of
them contain a ricidulously small amoimt of nourishment at the
price. It is vastly more expensive to rear a child upon one of
them than upon fresh or even condensed milk.

iJBER DIE CHEMISCHE ZUSAMMENSETZUNG DES
"SALZBREIES" VON BONITO ("SHIOKARA")

Von U. Suzuki, C. Yoneyama und S. Odake,
Landwirtschaftliche Facultdt, Kaiserliche Universit&t, Tokyo, Japan

Zur Bereitung des "Salzbreies" wird der Magen und Darm
des Bonitos vom inneren Inhalt befreit, gut gewaschen, fein
zerhackt und mit viel Kochsalz vermischt, so dass es einen
dicken Brei gibt. Die Leber wird auch manchmal dazu gemengt.
Man lasst nun den so bereiteten Brei wochenlang bei Zim-
mertemperatur stehen und riihrt ofters um. Es tritt dabei all-
m&hlich die Reifung ein; es entwickelt sich ein eigentiimlicher
Geruch und Geschmack, und von vielen Leuten, besonders
von Sakekennern wird der Artikel als Delikatesse mit Vorliebe
genossen. Die an der Reifung des Breies teilnehmenden Mikro-
ben sind bis jetzt nicht untersucht, und die chemischen Vorgange,
die wahrend des Reifeprozesses vor sich gehen, sind auch noch
nicht naher erforscht. Nur vermutet man, dass sie den bei der
"ShOyu" Bereitung auftretenden ziemlich ahnlich sind. Durch
Einwirkung von Mikroben und Enzymen werden verschiedene
Stoffe, besonders Eiweissstoffe, allmahlich gelost imd abge-
baut, unter Bildung von Peptonen und Aminosauren, die zum
Teii weiter desamidiert, oxidiert oder reduziert werden. Es
entstehen dabei verschiedene Sauren, Alkohole, Amine u.s.w.
Die Zusammensetzung des Breies ist deshalb sehr komphziert.
Es kommen bei verschiedenen Reifestadien verschiedene Stoffe
zum Vorschein.

Wir beschranken uns vorlaufig mit der Untersuchung der
stickstoffhaltigen Bestandteile des kauflichen, gereiften Breies.
Das von uns imtersuchte Material war aus Odawara bezogen.
Es war grau-rotlich-braun gefarbt und reagierte ziemlich stark
sauer. Die quantitative Bestimmung gab folgendes Resultat:

339

340 Original Communications: Eighth International [vol.

In 100 Teilen frischen Breies

Wasser 65.13

Trockensubstanz 34 . 87

In 100 Teilen Trockensubstanz

Organische Stoffe 30 . 06

Asche 69.94

Chlor 29.80

(Als NaCl berechnet) 49. 18

In lOOg frischen Breies

Gesamt-N als 100

Gesamt-N

1.735

100.0

Eiweiss-N

0.472

27.2

Org.-Basen-N . . . .

0.447

25.7

Ammoniak-N

0.131

7.6

Anderes N

0.685

39.5

Zur Isolierung der stickstoffhaltigen Stoffe wurden 4 Kilo
Brei aus gepresst. Der Riickstand wm-de drei mal mit war-
mem Wasser (40-50°) extrahiert. Die vereinigten Ausziige, die
schwach sauer reagierten, betrugen rund 9 liter. Sie wurden
mit 20% iger Tannin losung gefallt. Der Tannin-Niederschlag
(B) wurde abgesaugt und mit Wasser gewaschen. Das Filtrat
vom Tannin-Niederschlag wurde mit verdiinnter Natronlauge
verstezt, bis es schwach alkalisch reagierte. Es entstand dabei
eine flockige Fallung (C) in reichUcher Menge. Man saugte
davon ab, und setzte dem Filtrat viel Baryt zu, um das Tannin
zu entfernen, saugte wieder ab und nach dem Entfernen des
Baryts mittelst Schwefelsaure dampfte man bei niederem
Druck stark ein. Es schieden sich dabei Tyrosin, Leucin und
anorganische Salze aus. Aus heissem Wasser umkrystallisiert,
erhielt man zuerst 3 g Tyrosin und von der Mutterlauge desselben
2 . 1 g Leucin. Beide Aminosauren wurden nochmals fiir sich
umkrystallisiert und analysiert.

xviii] Congress of Applied Chemistry

341

Tyrosin:

0.1604 g Subst. gaben 10.7 c.c. N (16° 760 ttitti)

CJIiiNOs Ber

7.70

Gef

7 78

Leucin:
0.1719g Subst. gaben 15.7 c.c. N (14° 758 m.m.).

CaHisNOj Ber

10 07

Gef

10.71

Die Mutterlauge von Leucin und Tyrosin wurde mit Schwefel-
saure angesauert und mit Phosphowolframsaure gefallt.

A Der Phosphowolframsaure-Niederschlag

Die aus diesem Niederschlag dargestellte alkalische Fliissig-
keit, die freie Basen enthielt, lieferte nach starkem Einengen
im Vakuum keine Krystalle, so wurde sie mit Kohlensaure
gesattigt und mit Quecksilberchlorid gefallt.

a) Der Quecksilberchlorid-Niederschlag wurde mit Schwefelwas-
serstoff zerlegt, im Vakuum ein gedampft und mit Pikrinsaure
erwarmt. Nach dem Erkalten schieden sich 8.5 g fast reines
Lysinpikrat aus, welches aus heissem Wasser umkrystallisiert
und analysiert wurde.

0.1414 g Subst. gaben 22.0 c.c. N (13° 766 mm)
0.1491 g " " 0.2094 gCOs 0.0612 gHjO

0.4149 g " " 0.2533 g Pikrinsaure

342 Origina Communications: Eighth International [vol.

7?"

Pikriosaure

CaHuNzO! CeHiNaOi Ber

38.40
38.34

4.53
4.56

18.67
18.53

61 07

Gef

61.05

Im Kapillarrohr erhitzt, zersetzte sich das Pikrat gegen 247°
(unkorr.).

Das Platinchlorid-doppelsalz des Lysins waren hygroska^
pische goldgelbe lange Prismen. Es schmolz bei 205° (unkorr.).

Fiir die Analyse wurde es im Vakuum bei 100° getrocknet.

0 . 3014 g Subst. gaben 0 . 1055 g Pt.

Pt.

CeHuNjOj, HsPtCls Ber.
Gef.

3.500
35.00

b) Das Filtrat vom Quecksilberchlorid-Niederschlag wurde
nach dem Entfernen des Quecksilbers durch Schwefelwasser-
stoff und der Salzsaure durcli Silbernitrat, mit einem Uberschuss
von Silbernitrat und Baryt versetzt. Der braune Niederschlag
lieferte 1 .3 g Lysinpikrat.

Die Analyse des gereinigten Salzes gab folgendes Resultat:
0 . 1313 g Subst. gaben 21 . 3 c.c. N (20° 760min)

CeHuNaOa, C6H3N3O7

Bar.

18 67

Gef

18 56

c) Das Filtrat vom Silbernitrat und Baryt-Niederschlag
wurde in bekannter Weise mit Phosphowolframsaure gefallt.
Aus diesem Niederschlag erhielt man wieder 6 g Lysin pikrat.

0.1374 g Subst. gaben 22.3 c.c. N (21° 763 mm)
0.1526 g " " 0.2160gCOi! 0.0672gH2O

xvni] Congress of Applied Chemistry 343

CiHuN.O» CeHiNiOr Ber

38.40
38.60

4.53
4.89

18 67

Gef

18.55

B. Der Tannin-Niederschlag (Trytophan)

Der vom wasserigen Extrakt des Salzbreies durch Zusatz von
Tannin erhaltene Niederschlag wurde mit 3% iger Schwefel-
saure wiederholt verrieben. Ein Teil ging dabei in Losnng.
Man filtrierte nun vom unloslichen Riickstand ab und setzte
dem Filtrate viel Baryt zu, um damit Tannin und Schwefelsaure
wegzuschaffen. Das vom dabei entstandenen Niederschlag
abgesaugte Filtrat wurde mit Schwefelsaure angesauert und mit
Phosphowolframsaure gefallt.

a) Durch Zerlegung des phosphowolframsauren Nieder-
schlags wurde eine Fliissigkeit erhalten, welche schone Trypto-
phanreaktion gab. Wird diese Fliissigkeit mit Essigsaure
angesauert und mit einigen Tropfen Bromwasser versetzt, so
entsteht eine rot violette Farbung. Beim Schiitteln mit Amyl-
alkohol geht der Farbstoff in das letztere Reagenz iiber. Um das
Tryptophan zu isolieren, wurde die Fliissigkeit mit so-\riel
Schwefelsaure versetzt, bis sie 5% der Saure enthielt, imd mit
Hopkinischem Reagenz gefallt. Es enstand dabei eine weisse
flockige Fallung, die absesaugt, mit 5% iger Schwefelsaure
gewaschen und mit Schwefelwasserstoff zerlegt wurde. Beim
Eindampfen des Filtrats im Vakuum schied sich ein Teil des
Tryptophans krystallinisch aus. Die Hauptmasse blieb jedoch
amorph, so dass das gereinigte Tryptophan nicht zur Analyse
ausreichte.

Aus dem Filtrate vom Quecksilbersulfat-Niederschlag des
Tryptophans wurde eine Base als pikrinsaures Salz isoliert.
Dies geniigte auch zur weiteren Untersuchung nicht.

b) Das Filtrat vom Phosphowolframsaure-Niederschlag lie-
ferte, nach der Estermethode verarbeitet, eine kleine Menge
Leucin.

344 Original Communications: Eighth International [vol.

C. Der Tannin =und Natronlauge Niederschlag

Wie oben erwahnt, lieferte das Filtrat vom Tannin Nieder-
schlag (B) durch Zusatz von verdtinnter Natronlauge, wieder
eine reichliche Fallung, die eine nicht unbetrachtliche Menge
Basen enthielt. Um die Basen zu isolieren wurde der Nieder-
schlag mit 5% iger Schwefelsaure verrieben, wobei ein grosser
Teil in Losung ging. Die braune Fliissigkeit wurde nun mit
einem Uberschuss von Baryt versetzt, vom dabei entstandenen
Niederschlag abgesaugt, mit Schwefelsaure angesauert und mit
Phosphowolframsaure gefallt. Nach Zerlegung des phospho-
wolframsauren Niederschlags in bekannter Weise erhielt man
eine alkalische Fliissigkeit, die freie Basen enthielt. Diese
Fliissigkeit wurde nun mit Kohlensaure gesattigt und mit
Quecksilberchlorid gefallt.

p) Der Quecksilberchlorid-Niederschlag (Histidin)
Aus diesem Niederschlag erhielt man eine alkalische Fliissig-
keit, die sowohl starke Paulysche Reaktion, wie auch Biuret-
reaktion beim Erwarmen gab. Bei Zusatz von Pikrinsaure
wurde 0.5g Histidinpikrat gewonnen, welches aus heissem
Wasser umkrystallisiert, im Vakuum bei 100° getrocknet und
analj'siert wurde.
0. 1430 g Subst. gaben 27.8 c.c N (24° 759 mm)
0.1517g " " 0.2034gCO2 0.0457gH2O

CHtNaO:, CHaNsO,

Ber

Gef

37.50
37.56

3.13
3.35

21.91
21.77

Die Analyse stimmt also mit dem Histidin pikrat iiberein.
Der Schmelzpunkt war jedoch viel hoher als beim gewohn-
lichen Histidin pikrat, welches aus den Spaltungsprodukten des
Eiweisses dargestellt wird. Im Kapillarrohr erhitzt, wurde es
gegen 200° braun und zersetzte sich gegen 210° (unkorr.) unter
Schaumen. Es handelt sich wahrscheinlich um eine Isomeric
des Histidins. Wegen Mangel an Material konnten wir das
optische Verhalten nicht untersuchen.

b) Das Filtrat vom Quecksilberchlorid-Niederschlag des

xvni]

Congress of Applied Chemistry

345

Ilistidins wurde mit Silbernitrat und Baryt gefallt. Aus diesem
Niederschlag isolierte man eine Base als pikrinsaures Salz,
welches 1.5g betrug. Das Pikrat bestand aus rot braunen
blattrigen Krystallen mit dem Schmelzpunkt 225° (unkorr.).
Die Analyse gab f olgendes Resultat :
0.1171 g Subst. gaben 24.2 c.c. N (20° 760 mm)
0.1372g " " 0.1715gCO2 0.0496 gHjO

0.2504g " " 0.1919g Pikrinsaure.

Pikrinsatire

CiH.J>J.O. (CHiNiO:), Ber

34.18
34.09

3.16
4.02

22.15
23.64

72 47

Gef

76 63

Die Analyse stimmt also mit dem Arginindipikrat, nur ist
der Gehalt an Stickstoff und Pikrinsaure etwas hOher.

c) Das Filtrat vom Silbernitrat und Baryt-Niederschlag
(b) vrarde wieder mit Phosphowolframsaure gefallt. Der
Niederschlag lieferte 2.34g Lysinpikrat mit dem Zersetzungs-
pimkt 245° (unkorr.).

D. Das Filtrat vom phosphowolframsauren Niederschlag

Das Filtrat vom phosphowolframsauren Niederschlag wurde
nach der Estermethode verarbeitet, indem die Phosphowolf-
ramsaure und Schwefelsaure durch Baryt entfernt und der
Uberschuss vom Baryt mittelst Schwefelsaure beseitigt und im
Vakuum stark eingedampft wurde. Der zuriickgebliebene
Syrup MTirde nun mit absolutem Alkohol versetzt, mit trockenem
Salzsauregas gesattigt und in bekannter Weise in die freien Estern
der Aminosauren verwandelt. Nach fraktionierter Destination
der Estern, wurden die f olgenden drei Fraktionen erhalten :

Temperatur

Estermenge

Aminosauren nach
der Verseifung

2
3

bis 75° (20mm)
75-100° "
fiber 100° "

9.0g
10.5
7.0

3.0g

7.8

6.0

346 Original Communications: Eighth International [vol.

Fraktion 1 bestand aus Alanin. Aus heissem Wasser umkrye-
tallisiert, bildete es farblose Nadeln mit slissen Geschmack und
zersetzte sich gegen 270°. Fiir die Analyse wurde es im Vakuum
bei 100° getrocknet.

0.1553g Subst. gaben 21.3 c.c. N (17° 760 mm)
0.1530g " " 0.2248gCO2 0.1040gH2O

C3H7NO2 Ber

Gef

40.45
40.07

7.87
7.55

15.88
16.03

Fraktion II. bestand auch hauptsachlich aus Alanin, nebst
einer Kleinen Menge Prolin.

Analyse des Alanins:

0. 1476 g Subst. gaben 20.4 c.c. N (21° 751 mm)

C3H7NO2 Ber.
Gef.

15.88
15.62

Kupfersalz des Alanins:

0.2114 g Subst. gaben 0.0703 g CuO.

(C3H6N02)2Cu Ber.
Gef.

26.22
26.57

xviii] Congress of Applied Chemistry 347

Fraktion III. bestand zum grossten Teil aus Leucin, nebst
Alanin und Prolin. Zweimal aus heissem Wasser umkrystalli-
siert wurde das Leucin in ziemlich reinem Zustande erhalten.
Es schmeckte schwach bitter und zersetzte sich gegen 280°.

Analyse des Leucine:

0.1467g Subst. gaben 13.4 c.c. N (19° 762 mm)

C6H18NO2 Ber.
Gef.

10.07
10.56

Das Prolin wurde isoliert, indem die nach der Verseifung des
Esters erhaltenen Aminosauren mit heissem absolutem Alkohol
extrahiert wurden. Der vereinigte alkoholische Extrakt wurde
eingedampft und der Riickstand nochmals mit absolutem
Alkohol extrahiert. Nach dem Verdampfen des Alkohols
wurde das Prolin in bekannter Weise in das charakteristische
Kupfersalz verwandelt, welches in heissem absolutem Alkohol
Mich war.

Die Ausbeute an Kupfersalz betrug 1 . 03 g. Fiii- die Analyse
wurde das gereinigte Salz im Vakuum bei 100° getrocknet :

0.1613g Subst. gaben 13.8 c.c. N (22° 752 mm)

(C6H8N02)2Cu Ber.
Gef.

9.60
9.59

Ferner wurde das Vorhandensein von Glutaninsaure im
Filtrat des phosphowolframsauren Niederschlags durch ihren
charakteristischen faden Geschmack ausser Zweifel gestellt.
Wegen Mangel an Zeit haben wir diese Saure nicht isoliert.

348 Original Communications: Eighth International [vol.

Aus 4 Kilo Salzbrei des Bonitos wurden isoliert:

1. Lysin pikrat 18 . 14g

2. Histidin pikrat 0. 50

3. Tyrosin 3.00

4. Leucin 4 . 06

5. Alanin 10.80

6. Leucin + Alanin 4.00

7. Prolin Kupfer 1.03

8. Tr3Tptophan Vorhanden

9. Arginin di pikrat(?) 1 . 50

10. Glutaninsaure Vorhanden

ON THE CHEMICAL COMPOSITION OF "SAKE"

By T. Takahashi and Goro Abe
College of Agriculture, Imperial University, Tokyo, Japan

On the chemical composition of Sak6 many reports have been
made: Some authors write on the composition which is con-
tained originally in the fermented mash and others on the
constituents which have been derived from the vat or cask which
contain sak6. In the factories of sak6 the fermenting vat and
the cask which is employed for the transportation of sak6, are
made from cryptomeria japonica; and therefore certain compo-
nents of the material of the vessels must be present and dissolved
in 8ak6 Ch. Kimoto' reported on sughi-oil in 1903 and
N. Nagai^ and T. Kimura made some researches on its chemical
composition and found protocatechin, a phinon-like substance,
vanillin and a kind of terpene and they have proved the presence
of the same substances in sak4. The terpene, mentioned by
the above authors, was by K. Keimatsu' proved to be a sex-
terpene and he also added as a new component of cryptomeria
japonica a phenol-like substance, having reducing property.
In the same year Yamamoto and Ishikawa* made studies on the
same reducing substance as regards its influence in determining
the reducing sugar in sak6.

On the proper composition of sak6 K. Keimatsu' made a
report on the furfurol in sak6 and one of us' made research on
the contents of furfurol and found that in young sak6, or shortly
after the fermented mash is pressed, there was no furfurol or, if

■Bulletin of the College of Agriculture Tokyo Imp. Univ. vol. 4. page
403.

•In a speech made at the Tokyo Chemical Society (1904) .

•Yakugakuzasshi (Journal of the Pharmaceutical Society of Japan, 1905,
March.)

•do: Sept. 1905.

•do: Decemb. 1904.

•T. Takahashi: The Journal of the Agricult. Society of Japan. May, 1905.

349

350 Original Communications: Eighth International [vol.

present, it was a trace; while in aged sak6, i.e., after the storage
during summer, it was always present in it. In the same year
H. Nishizaki' made a quantitative determination of furfurol,
but arrived at somewhat different results from the writer.
Recently H. Ito made experiments on this subject" with about
111 samples of sak6 and arrived at the same conclusion as the
writer. One of us* has made a report in respect to the quantity
of fusel oil and pointed out that the quantity of the substance
must be examined when we classify sake as regards quaUty.
Moreover, a small quantity of methyl alcohol was proved to be
present in ordinary sake', and a somewhat evident quantity of
methyl-lactate in certain samples of sak4 was found by one of
usi".

H. Nishizaki made a report on free acids" and sugars'* of sakd
and mentioned that the latter consisted chiefly of glucose, while
K. Suda^' already sixteen years ago made experiments on the
contents of the sugar of sak6 and reported that the sugar con-
sists chiefly of maltose. It seems to the writer that both authors
may be right; because they used only a limited number of
samples and arrived at altogether opposite results. If they had
examined a wider range of sanaples they would probably have
foimd out that their results were one sided. K. Keimatsu"
and Shimizu reported on the presence of acetaldehyde, fusel
oil, succinic acid, lactic acid and acetic acid in sak6.

On animo-acids, however, no one has reported yet and we
therefore present here the results of OTir investigations about
amino-acids and other components.

The general chemical composition of the samples was as follows:

'Journal of the Phaxmaceutical Society of Japan. Novemb. 1905.

''Journal of Tokyo Chemical Society. Vol. 32, No. 7. He proved
directly, chiefly by anilUn acetate, but the quantity was too small; he has
distilled sak6 under reduced pressure and below 35°C neutralized sak6.

sT. Takahashi. Journal of the Agricu. Society of Japan. Apr. 1905.

»T. Takahashi. Bulletin of the Agr. CoU. Tokyo Im. Univ. Vol. 6. No. 4.

loT. Takahashi. Bulletin of the Agr. Coll. Tokyo Im. Univ. Vol. 7. No. 4.

"Journal of the Pharmaceutical Society of Japan. May, 1905.

'2do: May, 1906.

"do: Apr. 1890.

'•do: Decern. 1905.

xvm] Congress of Applied Chemistry 351

1). Total— N 0.1865 %

2). Protein— N.

a). Stutzer's method 0.0072 %

b). Rumpler's method" 0.00672%

c). Precipitate by Pb-acetate and Pb-oxide 0.00447%

(Bungener u. Fries. Zeit. f. d. ges. B. 1894.69.)
d). Precipitate by basic lead acetate 0.00435%

3). Non-albuminoid — N.

a). Ammonia— N. (Wurster's method) 0.00629%

b). Organic base— N 0.0598 %

c). Other— N. (chiefly amino-acids) 0. 1131 %

4). Esters (as acetic ester) 0.0457 %

5). Total acid (as succinic acid) 0.2666 %

a) . Non-vol. acid (as succinic acid) 0 . 2596 %

b). Volatile acid (by difference)

(as acetic acid) 0. 00715%

c). Volatile acid (determined in the distillate)

(as acetic acid) 0.0216 %

The protein nitrogen determined by Stutzer's and Rumpler's
method is always higher than that of the nitrogen found in the
basic-lead-acetate precipitate of lead-acetate and lead oxide;
because the former two methods always precipitate in a certain
degree a part of albumoses and peptones beside proper proteins.
For this reason therefore, we must assume the presence of albu-
moses and peptones in sak6; but we can prove directly the
presence of both substances. If we take 100-200 c.c. of sak6
and after removing the protein by basic lead acetate and from
this filtrate after removing lead by H2S and evaporating to a
small volume a suSicient quantity of Zu-sulphate is added to
suturaled the solution, acidifying with sulphuric acid, there will
be found a precipitate of albumoses. From the filtrate of albu-
moses, after removing zinc by H2S and condensing in a small

"Rttmpler; Deutsch. Zeit. Ind. 1898, 1729.

The difference between contents of nitrogen determined by Stutzer's method
and that of the precipitate made by lead-acetate is shown from the above table
to be 0,0027%, and this must be the least quantity of the nitrogen of albumose
and peptones.

352 Original Communications: Eighth International [vol.

volume, this 'will contain peptones in solution, which will be
proved easily by Biuret reaction.

(I) Organic Bases

10 litres of the samples were evaporated imder reduced pres-
sure and at 60° C. to a small volume, almost equal to i of the
original volume. After this operation the protein-substance
was removed by basic lead-acetate and researches were made
about bases according to Kossel's method; but of histidine only
a trace was found and the characteristic crystals of the chloride
were not obtained, giving Pauly's diazo-reaction (1904). Ar-
ginin was not found. Picrate of lysin, about 1.1 grms, was ob-
tained as fine needles and platy crystals, which melted at 230° C.
(uncorr.), so that it contained some impurities.

(II) MONO-AMINO-ACIDS

Ten litres of sak6 were used for the isolation of mono-amino-
acids after E. Fischer's well-known ester method.

a) Under the pressure below 20 m.m.
(Esters prepared from 10 1. sak^.)

First fraction 60°-92° C. 1 . 0 grms.

Second fraction 92°-150° C. 1 .5 grms.

Third fraction 150°-235° C. Trace.

b) Under the pressure below 18 m.m.
(Esters prepared from 10 1. sak^.)

First fraction, below 41° C. 0.6 grms.

Second fraction 41°-60°C. 1.0 grms.

Third fraction 60°-98°C. 3.0 grms.

Fourth fraction 98°-150°C. 4.5 grms.

From the first fraction of series (a), 0.2 grms of alanin, 0.2
grms of leucine, and 0 . 1 grms of prolin were obtained. In the
second series 0.2 gr. of alanin from first fraction and 1 g. of leu-
cine, trace prolin from third fraction, were obtained. The
Alanin obtained from first series was added to the same from
the second series and after purification analysis was carried out.
It was very sweet, having a melting point of 243-245° C. (uncorr.)

xviii] Congress of Applied Chemistry 353

and decomposed at the point with evolution of gas. The result
of analysis was as follows : —

Substance taken: — 0.972.
Nitrogen = 12 c.c. (at 15° C. 760 m.m.)
Calculated as C3H7NO2 N = 15 . 67%
Found N = 14.51%

Prolin was bitter in taste and its copper salts contained two
components, one of which dissolved in absolute alcohol, while
others do not, indicating the presence of active and inactive
prolin.

Leucin was bitter in slightest degree so that it was washed
with absolute alcohol. It gave a melting point of 290-293° C.
(uncorr.), changing in brown at 270° C. already. It decomposed
at the melting point with emission of gas. The analytical result
was as follows:

Substance taken : 0 . 1226 grms.

Nitrogen 10.7 c.c. (at 11° C. 758 m.m.)

Substance taken : 0 . 1102 grms.

Cou 0.2165 grms.

Calculated as CeHuNoa Found.

N = 10.85 10.46

C = 55.80 53.57

H = 10.07 9.73

The above result tells us that the substance was not pure.

Phenylalanin and glutannic acid were not found, but trace
of aepartic acid was found.

Leucinimid. (CijH2N202). After all esters were evaporated,
the residue was treated with acetic-ester, which dissolved a part
of it. When the dissolved part was evaporated, there remained
hexagonal or quadratic plate, which tasted very bitter. The
platy crystals- were dissolved again in acetic ester and after
evaporaton ether and ethyl-alcohol were added, but there were
found no crystals. So we could not prove the presence of
leucinimid.

354 Original Communications: Eighth International [vol.

Tyrosin. The presence of this substance was easily proved
by Millon's reaction by the filtrate from the precipitate of pro-
tein— substance of sak^. But for the isolation of this substance
we followed Brown" and Willer's operation, which they em-
ployed for the isolation of this substance from malt. Thus 1 grm.
of tsTosin was obtained from three litres of sake. While, as in
other cases, 10 litres of the sample was evaporated to a small
volume and after separation of protein, the filtrate free from
lead, was evaporated to almost one-fifth of the original volume
of sak^ and left standing over night in a cool place, there ap-
peared the characteristic silky crystals of tyrosin amounting
almost to 6 grams. The isolated tyrosin gave strong red colora-
tion by Millon's reagent, faint reaction of Pauly's'^ diazo-
reaction strong by Wurster's reaction" and Denige's reaction."
The analytical result was:

Substance taken: 0. 1048 grms.

Nitrogen 6.8 c.c. (at 10° C. 762 m.m.)

Calculated as CgHnNOa N = 7-.777%

Found N=7.79%

Cystin. This substance was proved very easily in the filtrate
which was obtained in removing the protein-substance of sak6,
but the quantity was too small to isolate it.

Tryptophane, (C11H12N2O2). This substance was obtained
from 110 c.c. of sak6 by Hopkins' and Cole's'"' method. The
crystals were platy and bright ; giving red coloration with bromine-
water and precipitated by phosphotungstic acid. A few crystals
were mixed with caustic potash and after fusing and subjected
to the dry distillation, pyroll reaction was observed in this dis-
tillate. However, the presence of tryptophane in sak6 is hmited
only to young sak6, or not aged sak^.

" Brown u Wilier. Woch. f. Brauerei, 1907. Nr. 11. S. 139.
" Beautiful red color by acetic acid and natrium-nitrite (c. f . Ph. 1. 1903
(1888).
IS Hoppe-Seyler's zeit. f. ph. ch. 42, 517 (1904).

1. Wine-red by form aldehyd and HiSO. (comptes rendus, 130, 583, 1900).
"Journal of Phys. 27, 418 (1902), 29, 451 (1903).

xviii] Congress of Applied Chemistry 355

On this fact one of us" has reported already. On the question,
that why this substance disappears in the aging process of
8ak6, H. Ito*' has made some observations and arrived at the
conclusion that the tryptophane in yoimg sak6 is assimilated
or rather decomposed by so-called aging yeast^: — ^WilUa anomala
varieties.

The Substances which dissolved in Ether

1.) Succinic acid.

Ten litres of sakd were evaporated to about 400 c.c. under
reduced pressure below 70° C. The syrup thus obtained was
extracted with ether, using Kumazama and Sudo's extraction
apparatus and dried. From ether extract, the ether was evap-
orated for a long time in sulphuric acid containing desiccator,
then there appeared long platy or mono-crinic prismatic crystals
in a brown colored syrup. The crystals were separated by
filtration and after washing very quickly with a small quantity
of cold absolute alcohol, there remained rather small quantities
of the crystals, which melted at 181°-182.5° C. (uncorr.).
The substance behaves very strongly acidic to Utmus, tasting
characteristic to succinic acid, and gave pyroU reaction when
subjected to Neuberg's prove (1900-1901). Such pure sub-
stance obtained from ten litres of sak6 was about 3 grams."*

From another 18 litres, 4.7 grams of the substance was ob-
tained as raw product, which was brown in color, so that it was
first neutralized with a 5% solution of natrium hydroxide, and
after evaporating to syrup, absolute alcohol was poured on there
appeared almost spontaneously fine crystals of natrium succinate.
The natrium salt thus obtained was washed repeatedly with
absolute alcohol. It was silky white and gave a brown precipi-
tate with ferric chloride.

" T. Takahashi. Journal of the Tokyo Chemical Society of Japan, Vol. 32,
No. 3, 1911. Also, Journal of the College of Agr. Tokyo Imp. Univ. Vol. 000,
No. 0.

"H. Ito. Journal of the Tokyo Che. Soc. of Japan, Vol. 32. No. 7, 1911.
Also, Journal of the College Agr. Tokyo Imp. Univ. Vol. 00, No. 00.

" T. Takahashi's report. Journal of the College of Agr. Tokyo Imp. Univ
Vol. 1, No. 3.

" The inactive form of zinc salt contains 18.17% of water of crystallization.

356 Original Communications: Eighth International [vol.

2). Lactic acid.

The syrup obtained after removal of succinic acid crystals
was neutralized with zn-carbonate to attain a thick pasty mass
from which, after standing overnight, the crystals of zinc lac-
tate were separated by "nutsche" and washed with absolute
alcohol. The salt thus obtained was treated with H2S to sepa-
rate zinc, and the colorless solution of free acid thus obtained
gave intense reaction of Ueffermann's proof. Zinc salt was re-
prepared from this pure lactic acid and after drying well in the
desiccator, the water of crystallization was determined, drying
several hours at 105-110° C. The water^^ of crystallization
amounted to 17.0%, almost equal to that of the inactive form
of lactic acid.

3). Tyrosol. OHC6H5CH2CH2OH.

For the isolation of tyrosol we followed Fe. Ehrlichi's^'
method. Ten litres of the sample were evaporated to almost
400 c.c. and after basifying with NaHCO^, ether extract was
made by Sudo and Kumagawa's extraction apparatus. The
yield was 4 grams as raw product. It was dissolved again in
absolute alcohol and decolorized with animal black. It be-
haved very strongly to Millon's reagent and diazobenzol sul-
fonic acid and faintly to ferric chloride and Denigfe's-Moerner's
proof. Tyrosol-di-benzoate prepared by the addition of benzoyl
chloride and natroulye, melted at 113° C. (upcorr.).

SUMMAEIES

The above statements are summarized below:

The jdeld of the observed substance from 10 litres of Sak6:—

1). GlycocoU Not found.

2). Alanin 0.2 grams.

3). Leucin 0.6 grams.

4). ProUn (active and racenic) 0.1 grams.

5). Phenylalanin Not found.

" Ber. I. Deut. ch. G XLIV, Heft I. S. 139-146, 1911, u Bioch. Zeit,
36 Band, 15 Heft. S. 477, 1911.
"F. Ehrlich, Ber. d. Deut. ch. Ges. 1911, XLIV, Heft I. S. 143.

xnv] Congress of Applied Chemistry 357

6). Glutamic acid Not found.

7). Aspartic acid Trace (?)

8). Leucin-imid Not found.

9). Tyrosin 6.0 grams.

10). Cystin Trace.

11). Tryptophane (only present in young

sak6) 1.0 grams.

12). Lysin 0 . 25 grams.

13). Ristidin Trace.

14). Arginin Not found.

15). Tyrosol 4 grams (raw products).

16). Succinic acid 3 grams (in the least).

17). Lactic acid (inactive) 2 grams (in the least).

18). Albmnoses and peptones present.

The yield of amino-acid is too small in comparison to the
nitrogen contents of amino-acids obtained from calculation.
The main cause is that the pressure, under which we have made
fractions was rather high (18-20 mm.), but beside this sakd
contains rather high percantage of carbohydrate, which makes
it diflBcult to isolate the esters. In conclusion the writer must
offer many thanks to his assistant M. Sato, for his faithful help
during this research.

(Abstract)

A STUDY OF THE COMPOSITION OF CIDER VINEGAR

MADE BY THE GENERATOR PROCESS
L. M. TOLMAN AND E. H. GOODNOW

Bureau of Chemistry, U. S. Department of Agriculture, Washington,

D.C.

This paper is the result of factory experiments on a very large
scale of the conversion of fermented cider into vinegar in the
ordinary commercial type of vinegar generator.

Experiments were carried on for a period of several months,
and some 40,000 to 50,000 gallons of hard cider were run through
a series of generators, samples being taken from the material go-
ing on to the generator and from the finished product, and analy-
ses made in detail to show the changes which took place. Work
was undertaken largely because the existing data published, re-
garding American cider vinegars was based wholly upon the analy-
ses of vinegars made in a small way, and fermented under the
slow fermentation process, which takes place in a barrel or cask.
It was found at the beginning of this investigation that the results
of analyses on this type of vinegar showed such a wide variation
that it was practically impossible to detect any forms of adulter-
ation. And it was concluded from our analyses of ciders existing
at that time that this wide difference in results was due to the
method of manufacture; and as the method of manufacture in this
country at the present time is practically confined to the generator
process, it was found that it was necessary to have data upon this
product.

Results of the investigation showed that vinegars made by the
generator process were practically as uniform in composition as
the cider from which they were niade, showing a very different
condition from that found to exist in the existing data on cider
vinegar.

359

THE MICROSCOPICAL EXAMINATION OF VEGETABLE

PRODUCTS AS AN ADJUNCT TO THEIR

CHEMICAL ANALYSIS

By a. L. Winton
U. S. Food and Drug Inspection Laboratory, Chicago, III.

In solving the problems of man and nature the analytical
chemist too often limits himself to chemical or physico-chemical
methods. He is an analj'tical chemist in the strict sense of the
word and not an analyst, which implies a man of broader training
and experience, utilizing the principles of other sciences as means
to his end. He turns his back on the methods of vegetable and
animal histology, physiology and bacteriology, asserting with
satisfaction that he is a specialist and as such must limit his field
of activity.

This attitude of the analytical chemist may be traced to a mis-
apprehension as to the province of a specialist. Such a worker
must be hmited only in the field of appUcation and not in training
or the methods employed. An occuhst, for example, Umits him-
self to defects of vision and diseases of the eye and allied organs,
but in order to properly carry out the work of his specialty he
must have broad medical training and be conversant with the
general principles of optics, bacteriology, chemistry and perhaps
other sciences. SpeciaUsts in other sciences, both pure and
applied, must also have good general training if they are to
achieve distinction in their limited fields; otherwise they are in
much the same position as the mechanic who, instead of mastering
his trade, learns to operate one machine, thus becoming a mere
automaton.

Botany and chemistry are generally considered incompatibles.
The student of chemistry sometimes takes up bacteriology as a
minor subject, but comparatively seldom studies advanced bot-
any, even though he intends to speciahze in food analysis, textile
chemistry, paper technology or some other subject dealing chiefly
" 361

362 Original Communications: Eighth International [vol.

with materials of vegetable origin. No physiological chemist
would think of pursuing his investigation of animal materials
without a working knowledge of animal anatomy, yet agricultural
and food analysts and others dealing with vegetable materials
too often limit themselves to a knowledge of chemical constitu-
ents, ignoring the relation of composition to histological structure.

This is most remarkable, since the methods of vegetable his-
tology, as well as of chemistry, are invaluable in solving problems
relating to the nature or constituents of foods, drugs, fibers and
other products of vegetable origin. Sometimes one line of
investigation alone is useful, sometimes the other, but often each
throws some light on the subject, and the corroboratory results
obtained by such widely differing means furnish an indisputable
chain of evidence.

Let us look more closely into the natm^e and relation of these
two applied analytical sciences.

Chemical analysis deals with chemical constituents; microscop-
ical analysis deals largely with the form of some of these constitu-
ents. Chemical analysis determines the amount of fiber, starch,
protein, oil, etc.; microscopical analysis determines the shape,
size, and other characteristics of the cells and cell contents.
Chemical analysis usually stops with the mere determination of
the amount of chemical constituents; microscopical analysis
goes further and names the particular product from which they
were derived. Chemical analysis answers a question only in
scientific terms; microscopical analysis, in terms which all can
imderstand.

In many cases, the best idea of a material is gained by following
out both lines of investigation. By chemical analysis we learn
the percentage of protein, fiber, starch, etc., but not the ingredi-
ents from which they were derved; by microscopical analysis
we learn the ingredients, but usually gain only an approjdmate
idea of their proportion. Given the results of both analyses,
we may often calculate with some exactness the percentage of
the different materials present.

If, for example, we find in a sample of wheat bran 11 instead
of 16 per cent, of protein, and 15 instead of 8 per cent, of fiber,
we know it is not pure bran but we do not know the adulterant;

xviii] Congress of Applied Chemistry 363

if we find corn-cob tissues under the microscope, we learn the
adulterant but not the amount. Knowing that the material is a
mixture of bran and ground corn-cob, and knowing the average
percentage of protein and fiber in both, we are in a position to
calculate from the results of the chemical analysis the relative
amounts of these ingredients.

Again, if we find in ground mace 40 per cent, instead of 20 per
cent, of fixed oil, we know it is not pure mace; if we find imder
the microscope a large amount of tissues of the Bombay mace,
a material worthless as a spice containing about 60 per cent, of
fixed oil, we learn the adulterant. Knowing all this, and knowing
the average percentage of oil in true mace and Bombay mace,
we have the data for calculating roughly the percentage of each in
the mixtiu-e.

Still again, if in a textile fabric we find a certain percentage of
organic fiber insoluble in boiling alkali, we know that the fabric
is not all wool. If under the microscope we identify this insoluble
fiber as cotton, we have found the missing link in the chain of
evidence.

In the analysis of complicated mixtures, we must often rely
entirely on microscopical examination. For example, chemical
analysis of a mixture of wheat, buckwheat and corn flours gives
us little information, and it is only after the characteristic starch
granules and tissues of each have been found under the microscope
that we gain a definite idea of the nature of the constituents.

Again, in the examination of paper, the microscope is our sole
dependence in learning the nature and approximate percentages
of the fibers employed, chemical analysis serving merely to
determine the kind and amount of sizing, coating and other non-
fibrous constituents.

Among some condimental cattle foods examined by the writer
some time since was one, the chemical analysis of which disclosed
but one proximate constituent, viz., common salt; the micro-
scope, however, disclosed linseed meal, corn meal, wheat feed,
mustard hulls, cocoa shells, malt sprouts, fenugreek and tiu-meric.
In such a case, dependence must be placed entirely on the micro-
scope, except for noineral ingredients.

Chemical analysis of another sample demonstrated the presence

364 Original Communications: Eighth International [vol.

of ground bone, carbonate of lime, iron oxide and free sulphur;
microscopical examination disclosed linseed meal, wheat feed and
charcoal. This is a striking example of a material in which half
the constituents (all mineral) can only be detected by chemical
analysis; the other half (all vegetable) by the microscope.

Many other equally striking examples of the interdependence
of these two applied analytical sciences might be cited.

The point now arises as to who is to carry on these two hnes of
investigation so different in details but so similar in purpose.

One plan is for a chemist to make the chemical analysis and a
botanist the microscopical examination. This plan has the
advantage that each can confine his attention to one specialty,
but it had the disadvantage that the close partnership between
the two, which is essential to the best results, outside of large
institutions, is both difficult and expensive. Such a division of
labor would usually be as impracticable as to divide the work
of a chemical laboratory between a chemist and a physicist, the
former conducting the precipitations and other chemical processes
the latter, polarizations, determinations of specific gravity,
refractive index and the like.

The rational plan is for one man to master both lines of research.
Such a man need not execute all the details, but he should be
thoroughly acquainted with them and should interpret the
results. We will call him an analyst, not a chemist or a botanist,
and his laboratory an analytical laboratory, not a chemical or
botanical laboratory. His equipment should consist of the
necessary apparatus for a wide variety of chemical work and
a complete microscopical outfit, including micro-reagents and a
set of standard specimens of economic seeds, roots, barks, fibers,
woods, etc.

But in order to have workers in this field, we must have
suitable courses of instruction in our schools of science. The
subject has a recognized place in many continental imiversities,
particularly in the schools of medicine, pharmacy and hygiene,
but outside of a few institutions, receives httle attention in
America.

The student who seeks to prepare himself for this field should
take both chemical and botanical studies. In chemistry, he

xviii] Congress of Applied Chemistry 365

should study the branches taught in a well-regulated chemical
course — elementary chemistry, qualitative and quantitative
analysis, organic and physical chemistry, and so on. In botany
he should take up successively elementary botany, systematic
botany (at least of the phanerogams) and vegetable anatomy and
physiology. These studies are all on the curriculum of every col-
lege and school of technology, although the student of chemistry
does not usually take all the botanical studies named. Without
a certain amount of botanical training, however, a chemist is
no more fitted to take up microscopical analysis than a botanist
without chemical training is fitted to work at quantitative
analysis.

After his preliminary studies in chemistry and botany, the
student is ready to take up a course in the methods for the
chemical and microscopical examination of the various raw
materials and of the products derived from them. This course
should be so arranged that the student will carry along his chemi-
cal and histological practice side by side, as he must do after-
wards in practical work. For example, in studying the cereal
grains, he should devote part of his time to the methods of deter-
mining water, ash (including ash analysis), protein, fiber, starch,
fat, pantosans, etc., and another part to a systematic study
of the starches and the histological elements of the bran coats
both in sections and in powdered form. In like manner, he should
take up a chemical and histological study of leguminous seeds,
oil seeds, spices, tea, coffee, cocoa, drugs, fibers, etc.

His work in the chemical laboratory should teach him not only
the strictly chemical methods but also the use of the polariscope,
the spectroscope and other physical apparat\is, and his microscop-
ical instruction should fit him not only to differentiate organized
forms but other characteristic elements, such as fat crystals,
mineral crystals, and the hke.

After such a course, he should be able not only to undertake
investigations in physiological or plant chemistry but also the
laboratory work of an official food department or a custom house,
a flour mill, a brewery, a sugar refinery, a candy works, a fruit
cannery, a drug mill, a textile mill, a paper mill, etc.

It is my firm belief that courses similar to that outlined should

366 Original Communications: Eighth International [vol.

be conducted in all our leading universities and schools of tech-
nology, and the student should be taught the use of the micro-
scope in conjunction with the balance in solving the analytical
problems which every day become more nimierous and intricate.

GENERAL INDEX

TO THE TWENTY-FOUR VOLUMES OF

ORIGINAL COMMUNICATIONS

Volume

Section

Analytical Chemistry.

Inorganic Chemistry.

Ilia

Metallurgy and Mining.

Illb

Explosives.

IIIc

SiUcate Industries.

Organic Chemistry.

IVa

Coal Tar Colors and Dyestuffs.

Industry and Chemistry of Sugar.

India Rubber and other Plastics.

Fuels and Asphalt.

Fats, Fatty Oils and Soaps.

Paints, Drying Oils and Varnishes.

Via

Starch, Cellulose and Paper.

VIb

Fermentation.

VII

Agricultural Chemistry.

Villa Hygiene.

Vlllb Pharmaceutical Chemistry.

VIIIc Bromatology.

Vllld Biochemistry including Pharmacology.

Photochemistry.

Electrochemistry.

Physical Chemistry.

XIa

Law and Legislation Affecting Chemical
Industry.

Xlb

Political Economy and Conservation of
Natural Resources.

ORIGINAL COMMUNICATIONS

EIGHTH INTERNATIONAL

CONGRESS
OF APPLIED CHEMISTRY

Washington and New York
September 4 to 13, 1912

SECTION Vllld: BIOCHEMISTRY INCLUDING
PHARMACOLOGY

VOL. XIX

ORIGINAL COMMUNICATIONS

EIGHTH INTERNATIONAL

CONGRESS
OF APPLIED CHEMISTRY

Washington and New York
September 4 to 13, 1912

SECTION Vllld: BIOCHEMISTRY INCLUDING
PHARMACOLOGY

VOL. XIX

The matter contained in this volume is printed in exact accordance with the manuscript
8ubmitted» as provided for in the rules governing papers and publicationB.

La mati^re de ce volume a Sti! imprimfte strictement d'aocord avec le manuacrit foumi et
lea regies gouvemant tous les documents et publications.

Die In dieaem Heft entiialtenen BeitreLge sind genau in X^ereinstimmung mit den ims
unterbreiteten Manuskripten gednickt, in Gemdssheit der fiir Beitrftge und Verlagaartikel
geltenden Bestimmungen.

La materia di questo volume e stampata in accordo al manosoritto presentato ed in base
alle Tegole que govemano i dooumenti e le publicazioni.

THE RUMFOBD PRESS
CONCORD-N-H'TT.S-A-

ORIGINAL COMMUNICATIONS

TO THE

EIGHTH INTERNATIONAL CONGRESS

APPLIED CHEMISTRY

APPROVED

BT THE

COMMITTEE ON PAPERS AND PUBLICATIONS

mVING W. FAY, Chairman
T. LYNTON BMGGS JOHN C. OLSEN

F. W. FREMCHS JOSEPH W. RICHARDS

A. C. LANGMUm E. F. ROEBER

A. F. SEEKER

SECTION Vllld. BROCHEMISTRY INCLUDING PHAR-
MACOLOGY

Executive C!ommitteb

President: John J. Abel, M.D.
Vice-President: William J. Gies, Ph.D.
Secretary: John A. Mandel, Sc.D.

Reid Hunt, M.D., Ph.D.
Thomas B. Osborne, Sc.D., Ph.D.

Sectional Committee

Carl L. Alsberg, M.D.
Silas P. Beebe, Ph.D., M.D.
Stanley R. Benedict, Ph.D.
Harold C. Bradley, Ph.D.
Russell H. Chittenden, Ph.

D., Sc.D., LL.D.
Albert C. Crawford, M.D.
H. D. Dakin, Sc.D.
Edward K. Dunham, M.D.
Charles W. Edmunds, M.D.
Otto Folin, Ph.D.
Harby-S. Grindley, Sc.D.
Robert A. Hatcher, M.D.
P. B. Hawk, Ph.D.
L. J. Henderson, M.D. .
Andrew Hunter, M.A.
Walter Jones, Ph.D.
Joseph H. Kastle, Ph.D.
P. A. Levene, M.D.
a. s. loevenhart, m.d.
John H. Long, Sc.D.
Graham Lusk, Ph.D., Sc.D.

and the Sectional

John J. Macleod, D.P.H.
Wm. deB. MacNideb, M.D.
John Marshall, M.D.,Sc.D.,
LL.D.
Albert P. Mathews, Ph.D.
S. J. Meltzer, M.D., LL.D.
Lafayette B. Mendel,Ph.D.
Frederick G. Novy, Sc.D.,
M.D.
Franz Pfaff, Ph.D., M.D.
Alfred N. Richards, Ph.D.
T. Brailsford Robertson,
Sc.D., Ph.D.
William Salant, M.D.
Philip A. Shaffer, Ph.D.
Torald Sollman, M.D.
A. E. Taylor, M.D.
Frank P. Underhill, Ph.D.
Carl Voegtlin, Ph.D.
Geo. B. Wallace, M.D.
Chas. G. L. Wolf, M.D.
Horatio C. Wood, Jr., M.D.
Executive Committee.

VOLUME XIX

SECTION Vllld: BIOCHEMISTRY INCLUDING PHAR-
MACOLOGY

CONTENTS

Alorich, T. B.

The Iodine Content of the Smali, Medium and Large Thyroid

Glands of Sheep, Beef and Hogs 9

ALBBEna, C. L. and Black, 0. F.

Biochemical and Toxicological Studies on PenicUlium Stolonif-

erum — Th<mi 15

Bbko, William N.

The Effect of Sodium Chlorid and Cold Storage upon Activities of

Proteolytic Enzymes 25

Black, 0. F. and Alsberg, C. L.

Biochemical and Toxicological Studies on PenicUlium Stolonif-

erum — Thom 15

Bloor, W. R.

Fatty Add Esters of Glucose 29

BuNZEL, Herbert H.

Qiuintitative Oxidase Measurements 37

Carles, P.

Les Phosphates et Le Sou De Froment Dans L' Alimentation

Animate 45

Carles, P.

Entretien Du Tissu Dentaire Par Une Alimentation Appropriee 49
Clark, Ernest D.

The Origin and Significance of Starch 55

Cooper, E. Ashley and Morgan, Gilbert T.

The Influence of the Chemical Constitviion of Certain Organic

Hydroxyl and Aminic Derivatives on their Germicidal Power 243

Crillat, M. a.

Influence Des Impuretis Gazeuses De L'Air Sur La VitalitS Des

Microbes 71

Crohn, Burrill B.

Experiences with Duodenal and Stool Ferments in Health and
Disease 73

6 Contents {vol.

Dubois, Raphael

Mecanisme Intime De La Production De La hwmiere Physiolog-

ique: Lvciferase, Lttciferine, Ludferesceine 83

Dubois, Raphael

Les VacuoUdes De La Purpurase et La Theorie Vaeuolidaire .... 91
Dubois, Raphael

Pharmacologie et Chimie Biologigue Atmolyse et Atmolyseur. ... 95
Ehhlich, Felix

Ueber Einige Chemische Beabstionen Der Mihroorganismen und
Ihre Bedentung fur Chemische und Biohgische Probleme 99

Eetzer, Lewis W.

The Chemical Changes Taking Place in Milk under Pathological
Conditions Ill

Fohtbscue-Bbickdale, J. M.

The Aryl Arsenates: their Pharmacology Considered from the
Experimental and Practical Standpoints 115

Foster, Lawrence F. and Hawk, P. B.

The Utilisation of Ingested Protein as Influenced by Undermas-
tication ("Bolting ") and Overmastication {" Fletcherizing ") 131

Fourneau, E. and Ochslin, K.

Chlorure de L'Adde Dichloroarsinobenzoique. Ethers des Acides
Benzarsineux et Benzarsinigue 136

Gebber, M. C.

Etude Comparie des Prisures de I'Amainte Phallmde et de I'Ama-
doumer — Relations Entre Les PrSsures Des Basidromycibtes et Des

Vigitaux Supirieurs 137

GrvENS, Maurice H. and Hunter, Andrew

Purine Catdbolism in the Monkey 149

Hawk, P. B., and Foster, Lawrence F.

The Utilization of Ingested Protein as Influenced by Undermas-

tication (" Bolting ") and Overmastication (" Fletcherizing ") 131

Hawk, P. B. and Howe, Paul E.

The Utilization of Individual Proteins by Man as Influenced by
Repeated Fasting 145

Herles, Franz

SchneUes Verfahren zurBestimmung der Hamsaure im Ham. ... 141
Howe, Paul E. and Hawk, P. B.

The Utilization of Individual Proteins by Man as Influenced by
Repeated Fasting 145

Hunter, Andrew and Givens, Maurice H.

Purine Catabolism in the Monkey 149

XIX

Contents 7

JowBTT, H. A. D. AND Pyman, F. L. and Remfhy, F. G. p.

The Relation Between Cliemical Consti.tUion and Physioloffical
Action as Exemplified by the Glyoxalines, Isoquinolines and Add

Anodes 153

lewis, Dean D. and Miller, Joseph L.

The Relation of the Hypophysis to Growth and the Effect of Feeding
Anterior and Posterior Lobe 231

LiNDET, M. L.

Sur Les Elements Mineraux Conlenus Dans La Caseine du Lait . . 199

Malvbzin, Philippe

La Question de I'Adde Sidfweaux Dans Les Vins Blancs 209

Marshall, C. R.

The Influence of Hydroxyl and Carboxyl Groups on the Pharma-
cological Action of Nitric Esters 211

Marshall, C. R.

The Pharmacological Action of BromnStrychnines 217

Maze, P.

Relations de la Plante avec les EUments FertUisants du Sol: Loi
du Minimum et Loi des Rapports Physiologigues 225

Menzb, G. a.

Srnne New Compounds of the Choline Type 229

Miller, Joseph L. and Lewis, Dean D.

The Relation of the Hypophysis to Growth and the Effect of Feed-
ing Anterior and Posterior Lobe 231

Morgan, Gilbert T. and Cooper, E. Ashley

• The Influence of the Chemical Constitutton of Certain Organic
Hydroxyl and Aminic Derivatives on their Germicidal Power 243

NicLoux, Maurice

Dosage et Moyen de Caraderiser de Petites Quantities d'Alcool
Methylique dans le Sang et les Tissus 259

Novi, Ivo

II Calcio e U Magnesia del CerveUo in Diverse Condizioni FisioU
ogiche e Farmacologiche 261

OCHBLIN, K. AND FOURNEAU, E.

Chhrure de I'Adde Dichtoroarsinabenzoique. Ethers des Acides

Bemarsineux et Benzarsinigue 1^"

PlCCININI, GuiDO M.

La Importama Fisiohgica del Manganese NeU'Organismo Ani-
mate... .. 263

Ptman, F. L. and Jowett, H. A. D. and Rbmtry, F. G. P.

The Relation Between Chemical ConstUution and Physiological
Action as Exemplified by the Glyoxalines, Isoguinolines and Add
Amides ^^

8 Contents [vol.

Reed, Howabd S.

The Enzyme Activities Involved in Certain Plant Diseases 265

Remfky, F. G. p. and Pyman, F. L. and Jowett, H. A. D.

The Relation Between Chemical Constitution and Physiological
Action as Exemplified by the Glyoxalines, Isoquinolines and Acid

Amides 153

Sauton, B.

Sur la Nutrition Minirale duBacille Tuberculeux 267

SCHtJLTZ, W. H. AND SEIDELL, AtHEKTON

Subcutaneous Absorption of Thymol from Oils 271

SCHULTZ, W. H. AND SEIDELL, AtHERTON

The Determination of Thymol in Dog Feces 281

Seidell, Atherton and ScHtFLTz, W. H.

Subcutaneous Absorption of Thymol from Oils 271

Seidell, Atherton and Schultz, W. H.

The Determination of Thymol in Dog Feces 281

Wolff, M. J.

Sur la Risistance de la Peroxydase a I'Ammoniaque et sur Son
Activation par Contact avec I'Alcali 287

THE IODINE CONTENT OF THE SMALL, MEDIUM

AND LARGE THYROID GLANDS OF SHEEP/

BEEF AND HOGS

By T. B. Aldrich

[From the Research Laboratory of Parke, Davis & Co., Detroit,

Mich.)

It is conceded by a majority, if not all writers on the subject
of thyroid therapy, that the thyroid gland (or its preparations)
to be physiologically active, must contain at least some iodine,
furthermore that this iodine to be of the greatest value thera-
peutically, must be combined or associated with some protein or
organic complex found in the gland. Presumably the iodine is
the more important constituent, the two, however, associated
or combined seem to give the best therapeutic results and to-day
the efficiency of a thyroid preparation is generally measured, or
should be by its iodine content. In fact for some time a number
of pharmaceutical houses have been putting out thyroid prep-
arations with a guaranteed percentage of iodine. Since then the
iodine content of a thyroid preparation is a measure of its thera-
peutic efficiency, it is desirable to select if possible those glands
which contain the most iodine, providing other factors are equal,
and from those animals, whose thyroids contain the most of
this constituent.

The following work was taken up with the object of deter-
mining the iodine content and thereby the therapeutic efficacy
of some thyroid glands, especially the small, medium and large
thyroid glands of sheep, beef and hogs by means of the method
employed by Hunter (Journal of Biolog. Chemistry 1909-1910,
VII, p. 321) which is very accurate and detects the presence of
very small amounts of iodine, that are incapable of being de-
tected by the older method of Baumann (J. Physiol. Chem-
1896, 21, p. 489; Ibid, 22, p. 1) which has been the method usu-
ally employed heretofore. (See Rigg's work J. of American
Chem. Soc. 1910) XXXII, p. 692; Ibid, 1909 XXXI, p. 710).

'The iodine content of some mixed sheep thyroids was also determined.

10 Original Communications : Eighth International [vol.

The glands received came from the Chicago stock yards, were
placed in Mason jars as soon as removed from the animals, sub-
sequently placed in cold storage and shipped packed in ice.
They were all received in the best possible condition. Six lots
were obtained as follows:

(1) Mixed sheep thyroids (Lots A and B).

(2) Small, medium and large sheep thyroids (Lot C).

(3) " " " " " " (LotD).

(4) " " " " beef " (Lot E).

(5) " " " " hog " (LotF).
In lots A, B and C the glands were not counted.

After freeing from superfluous tissue and weighing, the glands
were ground very fine, defatted and desiccated in the usual man-
ner and eventually reduced to a very fine powder by passing
through a 60 mesh sieve.

The following information obtained by one* of our staff from
a packer, relative to thyroids, may be of interest at this point.

(1) Sheep thyroids are subject to great variation in size. The
sex factor is not the determining factor for the size of the gland,
nor has the condition of nutrition of the sheep any decisive bear-
ing on the size of the gland. They run from the size of an almond
to the size of a lemon.

(2) Steers' thyroids are larger than cows', this variation in
size being a constant factor; size of gland varies again with the
condition of the animal. Well-nourished cattle have larger thy-'
roids than poorly fed ones.

(3) In hogs the thyroids vary little in size and present only
slight variation in general appearance.

The thyroids of cattle are removed after head has been sev-
ered; same is true of hogs. Cattle thyroids are often cut, those of
hogs not.

The following method of assaying the iodine, somewhat
abbreviated was employed. (For details see Hunter, The
Journal of Biological Chemistry 1909-1910, VII, p. 321).

Exactly one gram of the body was taken, placed in a nickel
crucible (125 c.c), 14 grm. of the following oxidation mixture
added (106 parts Sodium carbonate, 75 parts Potassium Nitrate,

XDc] Congress of Applied Chemistry 11

and 138 parts Potassium carbonate) and the two intimately mixed
by means of a nickel spatula. Over this was dusted 4 gm. of the
oxidation mixture. The nickel crucible was then heated over a
flame until contents of the same was perfectly white. This more
or less fused mixture was dissolved in water and brought into an
Erlenmeyer flask (800 cc). After cooling 35 cc. of Sodium
Hypochlorite solution was added, and while holding the flask
in a slanting position in cold water and agitating at the same
time, 65 cc. of 42|% phosphoric acid was added. The solution
should, after the addition of the acid, be colored slightly yellow
from the slight excess of chlorine liberated. The mixture was
then boiled briskly, a funnel with a short stem being placed in
neck of flask to avoid any loss. When all the free chlorin was
expelled, recognized by holding filter paper moistened with starch
solution containing Potassium iodine in the steam (blue color if
present), the flask containing now about 70-80 cc. was allowed
to cool and brought up to about 200 cc. by the addition of water.
To this cold solution 10 cc. of a 1% Potassium Iodide solution
was added, which causes the liberation of 6 times the amount of
iodine originally in the product to be assayed, according to the
following equation:

KIO, ■ HIOs + 10 KI + 11 HCl = 11 KCI + 6 H,0 + 12 I

This liberated Iodine is immediately titrated with a standard
solution of Sodium thio sulphate solution approximately N/200,
a few drops of starch solution being added toward the end of the
reaction.

The number of Cc. of Sodium thio sulphate used multiplied
by the iodine factor then divided by 6 gives the amount of iodine
in the original sample.

A blank test using casein, or some other body free from iodine
was made to insure the absence of iodine in the reagents, and the
usual precautions employed in analytical methods observed.

The following table gives the number (except in the lots A,
B and C), total weight of glands, average weight of glands and
iodine content in each lot with average iodine content and per-
centage in each gland, where the number of glands is known.

12 Original Communications: Eighth International [vol.

Sheep Thtkoids (Lots A and B)

Av. iodine in

each moist

gland

mg. %

No. of
Glands

Total Wt. of
glands

gms.

Av.
Wt. of
glands

gms.

Iodine %

(moist gland

tissue)

(A)
(B)

2925 (MX)i
5120 "

.032
.022

.025

Sheep Ththoids (Lot C)

(1)
(2)
(3)

6000 (S)
5200 (M)
5805 (L)

.027
.018
.01 J

.019

Sheep Thyboids (Lot D)

(1)

397 (S)

540

1.36

.044 1

.04

(2)

192 (M)

650

3.38

.028 [-0.028

.027

(3)

48 (L)

675

14.00

.015 J

2.1

.015

Beef Thyboids (Lot E)

(1)

98 (S)

620

6.32

.039]

2.47

.04

(2)

53 (M)

405

7.64

.030 1^0.036

2.40

.031

(3)

34 (L)

425

12.20

.038 J

4.70

.038

Hog Thyroids (L

3T F)

(1)
(2)
(3)

108 (S)
70 (M)
40 (L)

725
765
735

6.7
10.9
18.37

.054]

.048 [0.047

.041 j

3.6
5.2
7.5

.05

.047

.04

'The letters (S) (M) (L) and (MX) stand for small, medium, large and mixed
glands.

xDc] Congress of Applied Chemistry 13

The average iodine content of the mixed sheep thyroid glands,
Lots (A), (B), (C), and (D) where over 50 lbs. were employed,
is about . 025% while in some cases with selected glands 0.044%
has been obtained and .027 where relatively large amounts
(6000 gms.) were used.

The beef thyroids (Lot E) gave an average of .036% iodine,
and in selected cases nearly .04%. The hog .047% average, in
selected glands over .05% iodine.

It will be noted that the greatest variation in iodine content
in the different sized glands of the same animals, is in that of the
sheep where it varies from .01%-.027% in Lot C and from .015-
.044 in Lot D. This variation being no doubt due to the greater
prevalence of goiter in sheep.

Next to sheep the iodine content of the hogs' thyroids vary
the most .041-.054% while in the beef we have the least varia-
tion .030-.39%.

The hogs' thyroids^ contain the highest percentage of iodine,
with the small sheep glands in one case higher, in the other lower
than the mixed beef glands. Assuming the iodine content to be
a measure of the therapeutic activity, the mixed thyroids of the
hog are superior to the beef and the latter superior to sheep
thyroids, small sheep thyroids being about equal to mixed beef
thyroids.

Weight for weight the small glands of all the animals studied,
nearly without exception contain the most iodine (excepting
beef where the large and the small have nearly the same .038%).

In general the larger glands contain the most iodine and the
ratio of the iodine content of the small, medium and large glands
is approximately as follows:
In the sheep 2:3: 7
" " beef 1: 1:2
" " hog 3:4:6

The mixed glands arranged according to their iodine content
stand about in the following ratio :

'It is interesting to note that Baumann found very Utile iodine in pigs' and
hogs' thyroid glands; very much less than in beef and sheep.

14 Original Communications: Eighth International [tol.

Sheep Beef Hogs

5 7 9

From whatever standpoint we take we must conclude from
the above that the employment of either hogs or beef thyroids
for therapeutic purposes would be more rational than the em-
ployment of sheep glands, even if small selected sheep glands
are employed, thus eliminating the goiterous glands.

*It is a pleasure to thank Dr. Baeslack, of our staff, for looking after the
oollection of the glands and also for the information relative to the san>e.

BIOCHEMICAL AND TOXICOLOGICAL STUDIES ON
PENICILLIUM STOLONIFERUM — THOM

By C. L. Alsbebg and 0. F. Black
United States Department of Agriculture, Washington, D. C.

Whether molds or the products of their growth have an inju-
rious effect on animals is a question which has not yet been
conclusively settled. The literature contains many records of
alleged intoxications due to these fungi. Certain diseases of men
and domesticated animals have been attributed to this cause.
Though, obviously, the solution of this problem is urgent, few
serious attempts have been made to identify chemically the
alleged toxic substances. Chemical studies of this kind have
been undertaken in the Poisonous Plant Laboratory of the Office
of Drug Plant, Poisonous Plant, Physiological and Fermentation
Investigations, of the Bureau of Plant Industry, U. S. Depart-
ment of Agriculture. The present paper is the second of this
series of studies.

The genus Penicilium was chosen for study because, owing to
the mvestigations of Thom* it is now well systematized. The
necessity of using pure cultures of identified molds in an investi-
gation of this kind is obvious. Nevertheless, in most previous
investigations these factors have been neglected. Many of the
studies on molds deal with the action of unidentified mixtures
of molds on complex Substrata like maize or wheat. In many
instances in which pure cultures growing upon simple media were
studied, the identity of the species of mold employed can no
longer be established. This is due to the fact that these investi-
gations were, ordinarily, not conducted with the help of a trained
mycologist. Such help is absolutely essential, for the diffictilties
of distinguishing between species are ordinarily underestimated

'Thom, C. H., " Cultural Studies of the Species of PeniciUiuin," Bulletin
118, Bureau of Animal Industry, United States Department of Agriculture,
Washington, D. C.

16 Original Communications: Eighth International [vol.

by the clinical bacteriologist. In the present series of investiga-
tions the molds were isolated by Dr. Erwin F. Smith and iden-
tified by Dr. Chas. Thom. Without such aid these studies could
not have been undertaken.

In the first study of this series it was found that Penicillium
puberulum Bainier, produces a phenolic acid of the empirical
formula CsHio04, for which the name penicillic acid was sug-
gested.^ This acid gives a brownish red color with ferric chloride,
reduces Fehling's solution and yields a deep red dye when acted
upon by ammonium hydroxid. It is also somewhat toxic and
antiseptic. The lethal subcutaneous dose is from .2 to .3 grams
per kilo of body weight. It was not possible to identify peniciUic
acid with any known compound. In its general properties it
resembles very greatly certain of the lichen acids found in lichens.

In the present paper a similar study upon a closely related
organism, Penicillium stoloniferum, Thom, is reported. This
organism was isolated from a specimen of spoiled Italian maize
which was very kindly secured by Dr. C. H. Lavinder, of the
Hygienic Laboratory of the Public Health and Marine Hospital
Service while studying pellagra in Italy.

The examination of the specimen of Italian spoiled maize was
undertaken because as stated in a former publication^ it seems
to differ from American spoiled maize in its behavior toward the
ferric chloride reaction of Gosio. In Italy this reaction for pheriohc
substances is regarded as one of the most reliable tests for the deter-
mination of deterioration of maize. According to Schindler it
is not highly esteemed in the Tyrol. ' American spoiled maize
when tested by the method recommended by Gosio* only occa-
sionally gives the reaction. The color obtained in the few posi-

'Alsberg, C. L., and Black, O. F., " Biological and Toxicological Studies
upon Penicillium puberulum, Bainier. Proceedings of the Society for Experi-
mental Biology and Medicine, IX, p. 6, (1911)."

^Alsberg, C. L. and Black, O. F., "The Determination of the Deterioration
of Maize with Incidental Reference to Pellagra. Bulletin 199, Bureau of
Plant Industry, U. S. Department of Agricvdture, Washington, D. C, 1910."

'Alsberg, C. L. and Black, O. F. op. cit. page 27.

'Gosio, B.: Alterazioni Del Grantureo E Loro Profilassi. Page 35. Rome
1909. Tipografia Nazionale Di G. Bertero E C.

xix] Congress of Applied Chemistry 17

tive cases has always been red or brownish red, never violet or
green as described by Italian investigators. Recently the test
has been improved in this laboratory so as to render it more
delicate. The essential improvement in the procedure as now
conducted consists in extracting directly with chloroform. Fifty
grams of ground corn or meal are placed in a stoppered flask and
covered with chloroform. After two hours the chloroform is
filtered off and concentrated to a bulk of 10-15 cubic centi-
meters. This is transferred to a small separatory funnel or test
tube and covered with about 5 cubic centimeters of water con-
taining a trace of ferric chloride. If substances like penicillic
acid are present the characteristic color develops in the aqueous
layer. Conducting the tests in this way a greater number of
samples of obviously deteriorated maize show the reaction than
was the case with the old test. Nevertheless a positive result
seems to be less frequent in American maize than in Italian maize.

The Italian spoiled maize mentioned above gave an intense
ferric chloride reaction of a violet color. Moreover, when grown
on Raulin's medium it gave the same characteristic reaction. It
is certainly a remarkable fact that the first sample of spoiled
Italian corn examined gave the violet color described by Italian
authors, whereas no American sample has been found giving a
similar tint.

It was, therefore, decided to isolate, if possible, the substance
responsible for the ferric chloride reaction. For this purpose the
organism from Italian spoiled corn was grown on Czapek's
medium and on Raulin's medium. It was found that the organ-
ism grew more rapidly upon the latter. Therefore, for the prepa-
ration of material Raulin's medium only was used.

The substance responsible for the ferric chloride reaction was
isolated by the following procedure. The culture fluid and the
mycelium were transferred to an evaporating dish and rendered
weakly alkaline with sodium carbonate. The contents of the
dish were then heated to boiling and filtered hot. The mycelium
remaining on the filter was thoroughly expressed. It was then
again extracted with water, rendered weakly alkaline with sodium
carbonate. The combined extracts were evaporated to a small
bulk over a free flame and filtered hot. To the clear filtrate a
2

18 Original Communications: Eighth International [vol.

slight excess of hydrochloric acid was added. An abundant pre-
cipitate was produced which consisted of a mixture of needle
clusters and amorphous material. The precipitate was separated
by filtration and washed with cold water. After drying spon-
taneously it was extracted with hot toluene and the hot extract
filtered. Only the crystalline portion of the precipitate dissolved.
The amorphous dark brown material which remained on the
filter was discarded, for it did not give a color reaction with
ferric chloride. The toluene extract, on cooling and spontaneous
evaporation, precipitated in the form of needles, the material
giving the ferric chloride reaction. These were still sHghtly col-
ored but were finally obtained white, either by decolorizing with
boneblack in hot toluene solution or by dissolving in alcohol and
adding alcoholic potassium hydroxide to form the potassium salt
which is insoluble in alcohol. This salt was then washed free from
color with alcohol. From the potassium salt the free acid was
recovered in the form of white needles by dissolving the salt in
water and precipitating with hydrochloric acid.

The substance thus obtained consists of white needles with a
melting point of 140°, uncorrected. The name mycophenolic
acid is provisionally suggested for it. It is almost insoluble in
water but freely soluble in alcohol, in ether or in chloroform. It
is somewhat less soluble in benzene and only moderately soluble
in cold, though very soluble in hot toluene. With ferric chloride
it gives a violet color in aqueous solution, though its solubility
in water is not sufficient to render the color intense. In alcoholic
solution it gives a bright green color with ferric chloride. It does
not react with Millon's reagent. It does not give Lieberman's
reaction and could not be diazotized. It does not reduce Fehl-
ing's solution nor ammoniacal silver nitrate. It is fairly resistant
to sodium, ammonium and potassium hydroxide and hydro-
chloric, sulphuric and acetic acid, not being affected by boiling
in 10 per cent, solutions of any of these reagents. It does not
contain water of crystallization. Its salts of potassium and
sodium are very soluble in water. The former is soluble in dilute,
but insoluble in absolute alcohol. The latter is soluble in abso-
lute alcohol but may be precipitated in crystalline form by adding
ether. The salt of barium is only very slightly soluble in water

XIX

Congress of Applied Chemistry

and forms clusters of minute needles. The copper, lead and silver
salts are amorphous and insoluble in water. In characterization
of the substance the facts collected in Table I were ascertained by
analysis of the free acid, by titritation of the alcoholic solution of
the free acid with N/10 sodium hydroxide, using phenolphthalein
as indicator, and by the determination of the barium content of
the salt on ignition in platinum with sulphuric acid.

Table I. Analyses of Mycophenolic Acid

Weight of

Substances

Grams

CO2
Grams

HO2
Grams

C
Percent

H
Percent

BaSOi
Grams

Ba
Percent

N/lONaOH

Cubic
Centimeters

0.2316
0.2044
0.2494
0.1990

0.5419
0.4770

0.1315
0.1161

63.81
63.64

6.30
6.31

0.1256

29.65

11.53

Average

63.725

6.305

Calculated for CnHaoOe Carbon 63.74 Hydrogen 6.25 %

Found Carbon 63.72 Hydrogen 6.30 %

Calculated for Ba (CjHigOs) Barium 29.15 %

Found Barium 30.09 %

A molecular weight determination by the elevation of the boil-
ing point in chloroform solution gave the results in Table II.

Table II. Ebulioscopic Determination of the Molecular Weight
of Mycophenolic Acid

Weight of
substance

Weight of

Rise of

solvent

boiling

point

Grams

30.32

0.065°

30.32

0.060°

Molecular
weight

Grams

0.1641 30.32 0.065° 308

0.1578 30.32 0.060° 321

Average 314.5

Molecular Weight calculated for CivjHaoOj 320

Molecular weight found from titration 345.4

Molecular weight found from barium content of salt 328
Molecular weight found from boiling point elevation 314.5

20 Original Communications: Eighth International [vol.

The formula CnHzoOe may therefore be assigned to myco-
phenolic acid. It does not readily decompose carbonates at
ordinary temperatures. It is apparently a dibasic acid, or at any
rate, combines with two atoms of a monovalent base. Whether
the base combines entirely with carboxyl groups or with phenol
groups has not been determined. The acid seems to form two
series of salts. Presimiptive evidence on this point was obtained
by the following experiments.

0.2 grams of free acid were suspended in water and one equiva-
lent of potassium hydroxid added. Unfortunately, this was not
suflBcient to put the substances completely in solution, so that a
slightly greater quantity of the alkali had to be iised. This
solution was then treated with one equivalent of barium chloride.
On standing in the dessicator a crystalline barium salt formed.
This salt was evidently different from the normal barium salt
which is so insoluble that it precipitates at once. It was also of
different appearance under the microscope, consisting of a few
small needles in clusters, which apparently were the normal salt
and more abundant larger single needles, apparently the acid
salt. The presence of the normal salt in small quantities under
the conditions of the experiment was probably due to the fact
that an excess of alkali had to be used in dissolving the sub-
stances. The barium content of this preparation was deter-
mined, 0.207 grams yielding 0.0692 grams of BaSOi, equivalent
to a barium content of 20.2%.

Calculated for BaCCnHisOe 28.1 per cent.

Calculated for BaCCnHigOs)! 17.7 per cent.

Found 20.2 per cent.

Apparently, as shown by the microscope, the preparation
consisted of a mixture of two salts

It has not been found possible to identify the substances with
any known compound. In very many respects it resembles the
class of substances found in lichens and classed vaguely as lichen
acids. To find a substance of this class in molds is not surprising
since lichens are symbiotic forms composed of fungi and algae.
Mycophenolic acid also resembles very greatly a substance iso-
lated by Gosio from a species of Penicillium. The formula

xix] Congress of Applied Chemistry 21

CjHioOj calculated for the latter by Gosio is based on a single
combustion. As far as may be judged from Gosio's records, it
is probably not identical with mycophenolic acid, though resem-
bling it greatly. However, Gosio's characterization of the sub-
stance was based on a very small quantity of material, so that it
cannot be regarded as final. The chief points of difference be-
tween the substance described by Gosio and mycophenolic acid
are the percentage composition and the bahavior with ferric
chloride. Gosio's substance gives an intense blue color with
ferric chloride in alcoholic solution. Mycophenolic acid gives a
violet color in aqueous solution, while in alcoholic solution with a
trace of ferric chloride it gives a violet color which becomes
bright green on addition of an excess of the reagent.

In one particular mycophenolic acid resembles Gosio's sub-
stance but differs from penicillic acid. It is not toxic. Ten
milligrams were dissolved in water, with the aid of a little sodium
carbonate and injected subcutaneously into a mouse. No un-
toward effects whatever were noted. From penicillic acid,
furthermore, it differs in being present chiefly in the mycelium
in the early stages of growth. In old cultures it is found both
in the culture fluid and in the mycelium, perhaps because with
the gradual production of basic substances it is dissolved. The
question whether toxic phenolic substances are found in the cul-
ture fluid or only in the mycelium is one that has been much
discussed by students of pellagra. When the substances are
insoluble acids with soluble salts like mucophenolic acid, their
distribution is probably only a question of the reaction of the
medium. When the reaction is acid they will be found in the
mycelium as lichen acids incrust the lichen thallus. When the
medium contains available bases they will become more or less
dissolved in the medium.

With the advancing age of the culture mycophenolic acid
gradually increases in quantity until under the conditions em-
ployed in these experiments the maximum yield is obtained in
about two weeks. After that time the quantity present is
apparently constant. When grown in rectangular quart bottles
known in the trade as "long Blakes " turned on their sides in
order to have a large surface and charged with about 250 cubic

22 Original Communications: Eighth International [vol.

centimeters of culture fluid the yield at the end of about two
weeks averages per bottle about 0.07 grams of the crude acid.

Since P. stononiferum is found so commonly in the United
States it is not easy to understand why it is so rarely, if ever,
causes spoiled maize in the United States to give the ferric chlo-
ride reaction. The first explanation to present itself was that
the American organism might be a different strain or perhaps a
" physiological variety."

To solve this question Dr. Thom very kindly furnished a
specimen of his type culture. This was grown side by side with
the Italian organism. It grew rather more slowly than the latter
and there were slight differences in appearance. The cultures
gave a good ferric chloride reaction very similar in shade to that
given by the Itahan organism. When, however, the attempt
was made to separate mycophenolic acid from the cultures of the
American organism none could be found. In its place was found
a quite different substance or mixture of substances. As this
material has not yet been obtained in satisfactory crystalUne
form not much can at present be said of its properties.

The different biochemical behavior of the two strains from the
two continents is certainly suggestive. Whether these two strains
are really physiologically difJerent can not as yet be decided.
The American organism used is an old one, having been propa-
gated by Dr. Thom in the laboratory for a number of years.
Possibly this long artificial propagation has altered its behavior.
It is proposed to continue the investigation of this problem by
comparing the two cultures on hand with a number of new re-
cently isolated strains.

No extended physiological studies were undertaken on P.
stononiferum. A few observations were made incidentally. The
organism always produced alcohol as shown by applying the
iodoform test to the distillate. No quantitative determinations
were made but the amount of alcohol formed as judged by the
iodoform test seemed to be decidedly less than that produced by
P. puberulum. P. stoloniferum produces a small amount of oxalic
acid. To detect it the medium was concentrated to a syrup and
mixed with clean sand and plaster of paris. The hardened mass
was pulverized and extracted with ether in a Soxhlet apparatus.

xix] Congress of Applied Chemistry 23

The oxalic acid, identified by the melting point and insolubility
of the calcium salt, crystallized in the extract. Oxalic acid seems
to be present in somewhat larger amounts and at an earlier stage
of growth than in cultures of P pvherulum. Finally the myce-
lium of P. atoloniferum seems to be very rich in mannitol.

Summary

From cultures of Penidllium stoloniferum Thom obtained from
a sample of spoiled maize from Italy a new phenolic acid of the
formula C17H20O6 was isolated in crystalline form. It resem-
bles the lichen acids, is not toxic and is one of the substances
causing the ferric chloride reaction of Gosio in deteriorated maize.

THE EFFECT OF SODIUM CHLORID AND COLD STOR-
AGE UPON THE ACTIVITIES OF PROTEO-
LYTIC ENZYMES

Bt William N. Berg

Frmn the Dairy Division Research Laboratories, Bureau of Animal
Industry, Washington, D. C.

At low temperatures and in the presence of sodium chlorid
the activity of a proteolytic enzyme may be inhibited, if the
amount of enzyme is small. If the amount of enzyme is large,
proteolysis takes place rapidly and apparently is not interfered
with by the low temperatures and sodium chlorid. These obser-
vations were made during the course of some investigations
on the chemical changes taking place in cold storage butter. A
detailed account of these and related investigations is soon to
be published by the Dairy Division, Bureau of Animal Industry.

The Inhibiting Effect of Cold Storage and Sodium

Chlorid on the Activity of Galactase

in Buttermilk

Buttermilk obtained from a churning pasteurized or of unpas-
teurized ci'eam may contain galactase, a proteolytic enzyme
very similar in its general characters to trypsin. When butter-
milk is preserved with chloroform and kept at room temperature,
the galactase will slowly digest the proteins present. Some
quantitative data are given in a previous publication from this
laboratory.!

In buttermilk containing 18% of sodium chlorid placed in
cold storage, (at 0 F or minus 18 C.) for as long as nine months,
no proteolytic action was detected.

For the detection of proteolytic action, water soluble nitro-
gen was determined in the buttermilk before and after storage,
as follows:

'Rogers, L. A., Berg, W. N., and Davis, B. J., Circular 189, Bureau of
Animal Industry, 1912, p. 315.

26 Original Communications : Eighth International [vol.

Transfer 200 cc of the sample to a 500 ce volumetric flask.
Dilute with water to about 450 cc. Add 20 cc 10% acetic acid.
This will flocculate the casein. A cc more or less of the acid
will make no difference when sodium chloride is present. Make
up to the mark, filter on a 32 cm folded filter (S & S No. 595 or
588) and determine total water soluble nitrogen in two 200 cc
portions of the clear filtrate.

By this method, the results for water soluble nitrogen in
buttermilk containing 18% of sodium chloride and placed in
cold storage for 9 months, were the same, practically, before and
after storage. This indicates that the action of the enzyme
was inhibited under those conditions.

The Effect of Cold Storage and Sodium Chlobid on the

Activities of Pboteolytic Enzymes in Sterilized

Skimmilk

Digestive mixtures were prepared as follows: Skimmilk was
sterilized by heating for two hours at 94, 99 C. in a steam steri-
lizer. The skimmilk was quickly cooled to 35 C. and to three
3 liter portions in separate containers, 540 grams of sodium
chlorid was added, making the salt concentration approximately
18%. To one of these mixtures there was added 3 grams of pan-
cretin, dry, U. S. P., to the second, 3 grams of pepsin, dry, U.S. P.
and to the third 15 grams of a dry proteolytic enzyme prepara-
tion obtained from cultures of lactic acid bacteria which also
digested protein. The enzyme was precipitated from the
cultures (by L. A. Rogers) with alcohol in the manner usually
used for such preparations. The enzyme preparation was
tested before hand and found to liquefy gelatine.

Controls were likewise prepared, which differed from the
before described mixtures only in the fact that the enzyme
preparations were added to them while the shimmilk was near
the boiling point.

The mixtures were kept in sealed cans having a capacity of
1 liter. Each can contained 600 cc of the sample, and was kept
in cold storage for 9 months at 20 F (minus 7 C). Water soluble
nitrogen was determined in these mixtures before and after
storage by the method used for galactase.

xix] Congress of Applied Chemistry 27

It was found that under these conditions, the pancreat in
(trypsin) was very active, pepsin acted a little more slowly while
the bacterial enzyme preparation showed very little, if any
activity. The controls showed no change. In the trypsin
mixture 2-3 of the total nitrogen present was rendered water
soluble. In the pepsin mixture 1-3 of the total nitrogen was
rendered water soluble. When these mixtures were allowed to
stand at room temperature, further digestion took place. In
the trypsin mixture practically all the protein became water
soluble.

It is to be noticed that both pepsin and trypsin acted vigorously
in different portions of the same substrate.

However, the claim is not made that sodium chlorid does not
exert an inhibiting influence. Under certain conditions it does.
Experiments were made in this laboratory in the spring of 1909,
in which the speed of digestion of casein in several pepsin-acid
solutions was compared with that in the same solutions to which
20 grams of sodium chlorid had been added to 100 cc of acid solu-
tion. The presence of the salt almost completely inhibited the
action of the pepsin-acid during the experiment — 40 minutes'
digestion. It is of course probable that digestion would have
taken place had the digestion period been several months.
The method of comparing speeds of digestion was that de-
scribed by Gies.'

The results show that whether sodium chlorid does or does
not inhibit proteolysis depends upon the amount of enzyme,
to a very large extent.

'Berg, W. N., and Gies, William J., Journal of Biological Chem. Vol. 2,
pp. 489-546, 1907.

FATTY ACID ESTERS OF GLUCOSE

By W. R. Bloor

{From the Laboratories of Biological Chemistry of Washington
University, St. Louis, Mo.)

This paper is a preliminary report on the preparation and
properties of a new class of compounds — the fatty acid esters
of glucose. The interest of compounds of this type is threefold :
(1) the relationship which has been shown to exist between car-
bohydrates and fats in metabolism; (2) the possibility of the
natural occurrence of these and similar compounds; (3) the pos-
sible usefulness of such compounds in the study of fat metabol-
ism. A relationship between fat and carbohydrates in metabol-
ism has been repeatedly noted. In the absence of a sufficient
supply of available carbohydrate, as in starvation or severe dia-
betes, the fats are incompletely burned and the unbumed resi-
dues are excreted as B-oxybutyric acid and its derivatives, dia-
cetic acid and acetone. The fact has been crystallized in the
statement attributed to various investigators that " fats can burn
only in the fire of the carbohydrates." The acidosis may be
decreased or made to disappear if carbohydrates can be fed and
utilized.

That the condition may be reciprocal, i.e., that the fats, under
certain conditions may help in the metabolism of the carbo-
hydrates has, so far as I know, never been suggested but it seems
something more than a coincidence that the carbohydrate of
oats— the grain which has the highest per cent, of fat of all ordi-
nary grains — should be the best utilized by diabetics.

The nature of the relation of carbohydrates to the combustion
of fats has been the subject of many theories, the most reasonable
of which is that the glucose acts as a catalyser, either by furnish-
ing readily available oxygen, or by the formation of a compoimd
with the fatty acid which is more readily burned than the fatty
acid alone.

30 Original Communications: Eighth International [yol.

Glucose esters of the fatty acids have so far not been found in
nature. As may be seen from a study of the properties of the
compounds already prepared they are so much like the fats and
lipoids in their solubilities, etc., that they may well have escaped
detection. Compounds of galactose with fatty acids and other
substances are well known to occur in the brain substances
(cerebrosides).

Compoimds of this sort are of interest also because they may
be useful in solving the problem of the absorption and transpor-
tation of fats. If absorbed vmchanged they could be readily
recognized in the chyle and if injected into the blood stream could
be readily traced. Some work along this line has already been
done using analogous compounds— the mannite esters of the
fatty acids."

Glucose esters of butyric acid (di-butyrate) and stearic acid
(di-stearate) have been prepared by Berthelot^ by direct com-
bination at high temperatures. The jdeld was slight and his
description of the compounds is limited to their physical appear-
ance and two or three solubilities. Because of the small yield
and the instability of glucose at high temperatures this method
does not lend itself to the preparation of the compounds in large
quantity.

The synthesis, adopted depends on the action of the chlorides
of the fatty acids on glucose in solution in pyridin, the pyridin
acting both as solvent and catalyser somewhat as follows':

CeHsN

Cli

RC — CI =

CsHsN (intermediate addition

\ product)

C.R

'Bloor, Journal of Biological Chemistry, XI, p. 429.
^Berthelot Annals de Chemie et de Physique, (3) 60. 96. (quoted from
Beilstein Kandbuch der organischen Chemie, 3rd ed. vol. 1, p. 1049).
'Einhorn and Hollandt-Liebig's Annalen, 301. 95 (1898).

xixl Congress of Applied Chemistry 31

CI H

/ /

C.H.N -f- C.H„0« = RCo. OCHiiOs -^ GH.N

\ \

C.R CI

II
0

Process.

25 grm. of dry glucose is dissolved with the aid of heat in five
to ten times its weight of dry pyridin, the solution cooled, and an
equimolecular amount of the chloride of the fatty acid added in
small portions with cooling. The mixture is allowed to stand over
night, then poured into iced dilute sulphuric acid. The esters
separate and float on top, and are freed from the liquid (in the
case of the higher fatty acids) by filtering on a suction fuimel.
The mass is then boiled out several times with water, until it is
free enough from electrolytes to form a colloidal solution. It is
caused to separate from the colloidal condition by the addition
of sodium sulphate, let cool and the solid cake removed and dried.
The mixture is first fractioned with ether to separate the higher
esters and then with alcohol.

The following is a brief description of the compounds which
have been separated. Because of the great difficulty in making
the separations the data given are regarded as only approxi-
mately correct.

General properties of the esters.

The compounds all reduce Fehling's solution and are optically
active, the optical activity being less than that of glucose. They
form colloidal solutions with water (best made by pouring the
hot alcoholic solutions into water). They are readily saponified
by acids or by alkalies (watery or alcoholic). They do not fer-
ment with yeast. The presence of glucose was shown by sapon-
ification with alcoholic hydrochloric acid and preparation and
identification of the osazone. Some of the compounds possess
the property, in common with glucose, of forming sodium com-
pounds when treated in alcoholic solution with sodium ethylate.'
The compounds precipitate out as a gummy mass.

'Honig, Rosenfeld: Berichte der Deutsch. Chem. Gesellsch. 10. 1871.

32 Original Communications: Eighth International [vol.

Stearic add esters.

Yield from 20 gm. glucose and 30 gm. stearyl chloride — 35
gm. of crude esters.
Monostearate m.p. 110°. Specific rotation -5- 36.25°.

Slightly soluble in cold ether and alcohol; readily soluble in
hot alcohol and ether; separates slowly from cold alcohol.
The separation is hastened by the addition of ether.
Distearate m.p. 90-95°. Dextro rotatory

Slightly soluble in cold ether and alcohol; readily soluble in
them hot; separates quickly from alcohol on coohng.
Tristearate m.p. 60°. Specific rotation -f- 12.00°.

Soluble in cold ether; soluble in hot alcohol from which it sepa-
rates on coohng.

Lauric^addifisters.

Yield from 25 gm. glucose and 26 gm. lauryl chloride = 38 gm.
crude esters.

Monolaurate — well crystallized in shining rhombic plates, m.p.
110°. Specific rotation -^ 30°.
Slightly soluble in cold alcohol and ether; readily soluble in
them hot. The separation from alcohol in the cold is aided
by the addition of ether.
Dilaurate oc m.p. 55°. Specific rotation -H 21.8°.

Somewhat soluble in cold ether. Soluble in hot alcohol from
which it separates on cooling.
Dilaurate ^ m.p. 33°. Specific rotation -^ 30°.

Readily soluble in cold ether. Soluble in hot alcohol from
which it separates on cooling.
Butyric acid esters.

Yield from 25 gm. glucose and 12 gm. normal butyryl chloride
-H 3 gm. of mixed esters.

This synthesis is evidently not adapted to the preparation^ of
the butyric esters. The ester mixture agrees with Berthelot's'
description of the dibutyrate. " Very bitter liquid, somewhat
soluble in water, very easily in alcohol and ether." Strongly
dextro rotatory^ (h- 31°).

'Berthelot: loc. cit.

xix] Congress of Applied Chemistry 33

Animal experiments.

The material for the animaJ experiments was prepared from
the fatty acids of cocoa-nut oil after separation of oleic, palmitic
and stearic acids. ^ The fatty acid mixture used had a mean molec-
ular weight of from 200-210 and a melting point of 30°-36° C.
depending on the sample. Acid chlorides were prepared from
this product by the method of Krafft and Burger." The glucose
esters were prepared from the acid chlorides in the way described
above. The ester mixture so prepared and which was used for
the animal experiments had a m.p. of 41° but remained soft and
sticky at 30°. It was readily and completely soluble in cold
ether.

Specific rotation -7- 21°.

Feeding experiments.

The animal used for the feeding experiments was a cat, weight
2 k.

5 gm. of the ester mixture together with 25-30 gm. of lean meat,
5 gm. cotton seed oil and 2-3 gm. bone ash was fed every third
day. In the intermediate days and on the first two days of the
experiment the animal received the diet without the ester and
containing 1-2 gm. of wood charcoal in place of the bone ash. It
was hoped in this way to get sharply divided feces corresponding
to the feeding periods. As may be seen from the results it was not
possible in this experiment. Charcoal feces very often had a core
of bone ash feces and the two were otherwise so mingled that only
an approximate separation was possible. The feces from each
period were extracted with ether in a Soxhlet extractor for 3-4
hours and the extract examined polarimetrically for unabsorbed
esters.

Preliminary period. Two days.

50 c.c. ether extract, reading in 1 dcm. tube = -h 0.03° corres-
ponding to a weight of ester of 0.10 gm.

'For a description of the method of separation see Bloor: Jour. Biol. Chem,
XI, p. 421 (1912).

'Krafft and Burger: Berichte der Dutsch. Chem. Gesellsch. 17, 1378

(1884).

34 Original Communications: Eighth International [vol.

This figure was used as correction in the other periods.
First two ester periods (bone ash) extracted together.

65 c.c. of ether extract
Polariscope reading in 1 dcm. tube + 1.05°
Corresponding to a weight of ester of 3.40 gm.
Correction for blank 0.10 gm.

Corrected weight 3.30 gm.

First two control periods (charcoal)

28 c.c. of ether extract
Polariscope reading in 1 dcm. tube + 0.20°
Corresponding to a weight of ester of 0.28 gm.

Correction for blank (4 days) 0.20 gm.

Corrected weight 0.08

Summary of first two periods Ester fed 10 gm.

Ester recovered 3 . 40 gm.
Absorbed 6.60 gm.

Per cent, absorption 66%.

Third period (some diarrhoea)

Ester feces 98 c.c. of ether extract
Polariscope reading 0.50°
Corresponding to a weight of ester of 2.45 gm.

Corsection for blank 0.10 gm.

Corrected weight 2.35

Control feces 65 c.c. ether extract
Polariscope reading 0.03°
For third period Ester fed 5 gm.

Ester recovered 2.35 gm.
Absorbed 2.65 gm.

Per cent, absorption 53.0

Fourth period

Ester feces 120 c.c. of ether extract
Polariscope reading in 1 dcm. tube 0.12°
Corresponding to a weight of ester of 0.72 gm.
Correction 0. 12 gm.

Corrected weight 0.60 gm.

xix] Congress of Applied Chemistry 35

Control feces 40 c.c. ether extract
Reading in 1 dcm. tube 0.20°
Corresponding to a weight of ester of 0.40 gm.
Correction for blank 0.10 gm.

Corrected weight

0.30 gm.

For fourth period

Ester fed

5 gm.

Ester recovered

0.90 gm.

Absorbed

4.10 gm.

Per cent absorption 82%
Summary of experiments :

First two periods Absorbed 66%

Third period " 53% (diarrhoea)

Fourth period " 82%

Average absorption 67%

Injection experiments.

The material used for the injections was a colloidal solution of
the esters in water made by pouring a hot alcoholic solution into
water, filtering hot and boiling until the alcohol had evaporated
and the solution had reached a concentration of about 10%.
The milky suspension so obtained could be flocked out by the
addition of acids or of strong salt solutions, but could be diluted
with several volumes of normal salt solutions without separation
taking place for several hours. The ester mixture used in pre-
paring the solution was prepared for use by washing the ether
solution with dilute alkali until free from fatty acids, then sev-
eral times with distilled water.
Intraperitoneal injections.

The animal used was a young rabbit (Belgian haire) weighing
1.5 K. Two injections were made on succeeding days of (1)
5 c.c. of suspension containing 0.5 gm. of ester and (2) 10 c.c. of
solution containing 0.9 gm. of ester. The animal showed no bad
offects. Postmortem^ examination two weeks later showed that
part of one injection had lodged between the muscular coat and
the peritoneum. Microscopic examination of the cheesy mass

'I am indebted to Doctor W. S. Thomas of theTDepartmentTof Pathology
of this School for the postmortem examination.

36 Original Communications: Eighth International [vol.

showed it to consist practically entirely of leucocytes. Extrac-
tion of the substance with ether showed that the ester had dis-
appeared. Below the mass, the peritoneum was united to the
intestines by many adhesions. Scattered through the intestines
and also in the diaphragm and one edge of the liver were many
encapsulated masses of the same nature. Except for the above
all organs were normal.

Intravenous injections.

Made on rabbits.

Experiment I. Large (3 k.) rabbit with light brown spots.
Tap c.c. of solution containing 1 gm. of ester was injected into
the lateral ear vein during about 30 minutes. After the injection
the animal appeared normal and was put back into the cage. Ten
minutes later (about 40 minutes after beginning injection) it was
down and kicking convulsively and one minute later respiration
had ceased, although the heart continued to beat for a short
time longer. Autopsy showed a marked injection of the vessels
on the left side of the pons. Otherwise no abnormality.

Experiment II. Young rabbit weight 1.5 k. (the same one as
was used for the intraperitoneal injections above). First injec-
tion— 10 c.c. of solution containing 0.8 gm. of ester injected into
the lateral ear vein, the injection lasting fifteen minutes. After
the injection the animal behaved normally and showed no imme-
diate bad effects; nevertheless although fed liberally it rapidly
lost weight during the next few days. Second injection, five days
later — 6 c.c. of solution containing 0.9 gm. of ester injected into
the median ear vein, the injection lasting 15 minutes. After
the injection the animal appeared dull and hstless. No other
signs were noted for the next three hours but it died during the
night.

Discussion of the animal experiments.

The feeding experiments show that the glucose esters are
quite well utilized in the intestine. The injection experiments,
although too few in number to allow accurate deductions, indi-
cate that the substance is probably not well borne when injected
either intraperitoneally or intravenously. In the peritoneum
it seems to act as an irritating foreign body while intravenously
even if we regard the death of the first animal as an accident
its effects on the animal are injurious.

QUANTITATIVE OXIDASE MEASUREMENTS

By Heebeht H. Bunzel'

U. S. Dept. of Agric, Wash., D. C.

The very voluminous literature on the r61e and importance of
oxidizing enzymes in many vital processes of plants and animals
makes a thorough study of their behavior, function, and distribu-
tion necessary. They play an important part in certain patho-
logical conditions, and in numerous industrial, and agricultural
problems. As specific examples may be mentioned the work
done by Woods, in the Bureau of Plant Industry, on the mosaic
disease of tobacco, the work of Palladin and his school on the
respiration of plants, and the casual relationship between the
oxidases and color production as shown for plants by Palladin,
and for animals by Gortner. They also play an important part
in the darkening of tea, and the manufacture of Japanese lacquer.

Nearly all of the experiments made thus far have not been
carried on quantitatively because of the lack of satisfactory
methods.

The method described in this paper is based on oxygen absorp-
tion. For this reason a constant temperature is essential. The
apparatus in which the oxidations are carried out is shown in
the text figure. Eight cubic centimeters of the solution of the
substance to be oxidized are measured in the pipette G and allow-
ed to run into the compartment B. The plant juice, the oxidiz-
ing power of which it is desired to study, is measured in pipette
F and run into compartment A. Basket H holds 1 cc. of normal
sodium hydroxide to absorb the carbon dioxide formed in the
process; M is a manometer charged with mercury to indicate the
pressure within the oxidase apparatus. The whole apparatus is
clamped to a specially constructed shaking machine. In the

'From the Bureau of Plant Industry, U. S. Department of Agriculture,
Office of Drug Plant, Poisonous Plant, Physiological and Fermentation Inves-
tigations.

Original Communications: Eighth InternaMonal •! [vol.

Apparatus for Measuring Oxidase Action

air-thermostat the temperature is brought to 37° C. and mam-
tained at that point to within 0. 1 ° throughout the experiment.
Half an hour after the temperature of 37 ° is reached, all stop-
cocks but one are closed, and the shaking machine set in oper-
ation. The plant juice mixes with the oxidizable material and
the reaction begins. From time to time the shaking is inter-
rupted and the manometer is read. In the course of several
hours the oxygen absorption is completed, as indicated by no
further change of pressure within the flask. The ultimate
reading expresses the oxidase content of the juice or extract with
respect to the particular substance used. As a unit an oxidase
solution is chosen of such a strength that one liter of it will be
capable of bringing about the consumption by pyrogallol of the
equivalent of one gram of hydrogen.

xcc] Congress of Applied Chemistry 39

Hitherto pyrogallol, tyrosin. hydrochinone, guaiacol, benzidine,
and alphanaphthol have been investigated. The concentration
of the material to be oxidized has no effect on the end result pro-
vided it is used in excess. The carbon dioxide produced is
absorbed by the alkali in the basket and may be determined at
the end of the experiment by means of a special apparatus
devised for the purpose. The result obtained is directly pro-
portional, or at least nearly proportional, to the concentration
of the oxidase present.

An application of the method to potato juice is given in Table
1. In each one of the experiments 8 cc. of a 1% Pyrogallol
solution and 2 cc. of potato juice were used.

Table

Time of Manometric

Reading

Manometric Reading

P. M.