T h e Nutrient Content of High and
Low Quality Fresh Eggs. I. Total
Solids, Total Nitrogen, 1(—)
and Tryptophane
L. W. CHARKEY, ELIZABETH DYAR AND H. S. WILGUS, JR.
Colorado Agricultural Experiment Station, Colorado A. and M. College, Fort Collins, Colorado
(Received for publication June 21, 1947)
T HE apparent difference in quality
of the albumen of freshly laid eggs is
yet not adequately explained. Cole (1938)
reported that this difference in quality is
related to the size of the goblet cells of the
magnum of the oviduct. Conrad and Scott
(1942), however, were unable to confirm
this finding; they pointed out that their
observations were statistically non-signifi-
cant although based on a larger number of
birds (24 and 26, whereas Cole had used
only seven in each group).
It appears likely that the ability of a
hen or pullet to lay eggs of a given albu-
men quality is an inherent characteristic.
In any case a promising approach to the
problem of egg quality would seem to be
a study of the egg itself, aimed at the
determination of chemical or physico-
chemical differences existing between
eggs of high and low albumen quality.
It has been generally accepted that the
albumen quality of eggs is related to the
content and/or condition of mucin in the
egg white (Sharp, 1937; Conrad and Scott,
1939, 1942; Balls and Hoover, 1940).
Accordingly Conrad and Scott (1942)
studied the relationships between albu-
men quality and mucin content, and some
factors which might affect the colloidal
Scientific Journal Series Paper 247, Colorado
Agricultural Experiment Station.
626
state of the mucin. They were unable to
establish in these terms a satisfactory
explanation for variation in egg quality
and concluded that "something other than
the factors studied has a great influence
in determining the apparent quality of
a freshly laid egg."
Thus a more general study of the
chemical composition of eggs appeared to
be justified. The present report is a com-
parison of the total solids, total nitrogen
and 1(—) tryptophane in the separated
fractions of eggs of high and low albumen
quality.
EXPERIMENTAL
In order that a continuing supply of
fresh eggs of both high and low quality
would be available for analysis, two
groups of hens were selected from the
Colorado A &M College Poultry Hus-
bandry Department flock of New Hamp-
shire hens. The selections were based on
the albumen quality of eggs produced, as
determined by appearance, egg dimen-
sional measurements, and odor. The hens
were leg banded, and all eggs were col-
lected by trapnest so their source was
known. Once the characteristics of the
individual hens had been determined, the
identity of the two groups was maintained
only on the record of data. All birds were
afforded the same diet and treatment by
 NUTRIENT CONTENT OF FRESH EGGS
being housed and managed as a single
group.
The eggs were collected shortly after
being laid and were held at 10° C. until
examined the following day. Each egg
was then subjected to several measure-
ments to determine quality, although it
could be roughly assessed in advance by
referring to the group to which the hen
belonged. Eggs from the two groups were
removed from the refrigerator, allowed to
come to room temperature, weighed,
opened on a flat, level glass surface, and
immediately measured and observed for
odor or abnormalities.
Eggs were classified as high, medium,
or low albumen quality according to the
height of the firm albumen. Eggs whose
firm albumen measured more than 8.5
mm. in height were called high quality.
Those whose firm albumen measured less
than 6.5 mm. in height were classified as
low quality. The ones with firm whites
falling between these limits were medium
and were not used in these experiments.
All eggs were divided into the three main
fractions—yolk, firm albumen, and com-
bined inner and outer fluid albumen. The
chalaziferous membrane was discarded.
In order to have direct comparisons be-
tween high and low quality, the frac-
tions from an egg of high quality
and one of low quality were sampled
simultaneously and assayed simulta-
neously (except for total nitrogen deter-
minations) throughout the period of the
study.
Total Solids Determination
Because of the consistency of fresh egg
substance, the removal of moisture with-
out gross decomposition or loss of sample
presents a practical problem which is
best solved by using reduced pressure in
an apparatus permitting observation of
627
samples during drying. Since the methpd
used is a modification of the standard
A.O.A.C. procedure (1940) with condi-
tions more preceisly defined, it is given
here in detail:
The weighed samples, in dishes with the lids off-
set, resting on the edges of the dishes, were placed
in a vacuum desiccator which in turn was placed in
a drying oven at 70°C.The desiccator was connected
through openings in the sides of the oven to an
aspirator and by a second tube to a Milligan wash
bottle containing concentrated sulfuric acid. Be-
tween the desiccator and wash bottle a side tube was
provided so that rapid air flow could be provided at
only slightly reduced pressure at the start of the
drying period.
As rapidly as could be done without danger of
loss of sample by boiling over, the airflowand pres-
sure were gradually reduced by constriction of the
inlet tube. This in practice required about two hours.
The air flow was reduced finally to a few cubic
centimeters per minute, all of which was made to
pass through the sulfuric acid in the wash bottle by
complete closure of the side tube. This reduced the
pressure within the desiccator to 30-35 mm. Hg.
These conditions were maintained for 20 hours 4:1
hour, after which the samples were allowed to cool
in another desiccator and were weighed.
Total Nitrogen Determination
The dry residues from the total solids
determination were transferred (not quan-
titatively) to tared ashless filter papers,
re-weighed, folded into the papers and
secured by loops of pure copper wire.
Total nitrogen was determined directly
on these samples by the standard
A.O.A.C. macro Kjeldahl procedure
(1940).
Determination of l(—)tryptophane
1(—) tryptophane was determined by
the method of Woolley and Sebrell (1945)
using Lactobacillus arabinosus as the test
organism. Samples were prepared for
assay by a modification of the pepsin-
trypsin procedure of the above authors,
as follows:
Flasks containing weighed aliquots of
628 L. W. CHARKEY, ELIZABETH DYAR, AND H. S. WILGTJS, J R .

the fresh egg fractions for microbiological
assay were immersed in boiling water
(95°C.) until coagulation was complete,
as judged by solidification and the ap-
pearance of opaqueness. This required
about 60 seconds. Longer heating results
in further changes in which a translucent,
very tough collodion-like material is
formed. This condition is undesirable since
it hampers subsequent maceration of
samples. This heating and coagulation
procedure was included as a means of
Following digestion with pepsin, 0.3
gram of dipotassium phosphate and 0.3
gram of sodium acetate trihydrate were
added and the pH was adjusted to 4.6
with sodium hydroxide and hydrochloric
acid. Twenty mg. of takadiastase were
added as an aqueous suspension contain-
ing five mg. per ml. After the addition of a
few ml. of toluene, each flask was shaken
gently and incubated at 40°C. for 24
hours. The takadiastase digestion was
included in the procedure because it was

TABLE 1.—Content of total solids, total nitrogen, and l{—)tryptophane in

high and low quality egg fractions

Fluid albumen Firm albumen Yolks

high q. low q. high q. low q. high q. low q.

Total solids—percent 13.32 11.91 12.54 11.83 52.92 52.88

(24) (24) (24) (24) (25) (25)
Total nitrogen—percent
A. Wet basis (calc.) 1.81 1.62 1.71 1.58 2.60 2.60
B. Dry basis 13.62 13.54 13.60 13.35 4.92 4.87
(23) (23) (22) (22) (23) (23)
K~^tryptophane—mg./g.
A. Wet basis 2.06 1.48 1.78 1.59 1.80 1.72
(15) (15) (16) (16) (14) (14)
B. Dry basis (calc.) 15.88 13.11 14.22 13.13 3.40 3.25

minimizing the effect of the antitryptic
principle of egg white reported by Harte
(1945) to interfere in both raw and dried
egg white with in vitro enzymatic pro-
teolysis.
After the addition of 25 ml. of 0.1 N
hydrochloric acid and loosening of the ad-
herent coagulum by means of a policeman,
each sample was macerated by means of a
high speed cutting stirrer, the blades of
which could be passed into the digestion
flask, thus avoiding transfer of the con-
tents. Five mg. of pepsin (1:3000) were
added as a solution containing one mg.
per ml. in 0.1 N hydrochloric acid. Each
flask was shaken gently, stoppered with
cotton, and incubated at 40°C. for 20 to
24 hours. No toluene is necessary and none
was added, since it appeared to interfere
with the action of pepsin.
desired to assay the final preparations
for certain vitamins, as well as for trypto-
phane, and it was believed that proteolyt-
ic enzymes might fail to cleave certain
linkages by which vitamins are bound in
natural materials. The effectiveness of
takadiastase as used by Cheldelin el al.
(1942) in preparation of samples for vita-
min assays is well established.
Following digestion with takadiastase,
the pH was adjusted by means of sodium
hydroxide to 8.6. Six mg. of trypsin
(1:100) were added as an aqueous suspen-
sion containing one mg. per ml. After
addition of a few ml. of toluene when
necessary, each flask was shaken gently
and incubated at 40°C. for 16 to 20 hours.
After the above serial enzyme treat-
ments, yolk samples were extracted twice
with 25 ml. portions of ether. All samples
 NUTRIENT CONTENT or F R E S H EGGS 629

were then adjusted to pH 5.5 and steamed
for 15 minutes to drive off ether and tolu-
ene. The steamed samples were filtered
through Celite with suction, and the fil-
trates were diluted to convenient volumes
for assay.
The data were examined statistically
by Student's pairing method (Snedecor,
1940) after elimination of all data left
unmated by loss of samples. Table 2
states the degree of statistical signifi-
cance of differences found which are of

TABLE 2.—Summary of statistical comparisons: total solids, total nitrogen and ly—^tryptophane
content of high and low quality egg fractions

Values compared Means sSficance

Total solids—percent
Fluid albumen, high vs. low quality High 13.32 >999:1
Low 11.91

Firm albumen, high vs. low quality High 12.54 >999:1

Low 11.83

Yolk, high vs. low quality High 52.92 Not Sig.

Low 52.88

High quality eggs, fluid vs. firm albumen Fluid 13.32 > 99:1
Firm 12.54

Low quality eggs, fluid vs. firm albumen Fluid 11.91 Not Sig.
Firm 11.83
Total nitrogen— :nt (dry basis)
Fluid albumen, high vs. low quality High 13.62 Not Sig.
Low 13.54

Firm albumen, high vs. low quality High 13.60 >999:1

Low 13.35

Yolk, high vs. low quality High 4.92 Not Sig.

Low 4.87
/(—) tryptophane- f./g. (dry basis)
Fluid albumen, high vs. low quality High 15.88 >999:1
Low 13.11

Firm albumen, high vs. low quality High 14.22 ca. 16:1
Low 13.13

Yolk, high vs. low quality High 3.40 Not Sig.

Low 3.25

RESULTS AND DISCUSSION interest and of possible value in the expla-

Table 1 shows a summary of the find-
ings, containing the average values for
each class of sample. Numbers in paren-
theses indicate the number of individual
measurements represented by each aver-
age value shown. Since these data may
be considered from different viewpoints,
they are given on both the wet and the
dry basis.
nation of variation in egg quality. All
statistically significant differences found
are listed.
It is noteworthy that yolks from eggs
of high albumen quality were not found
to differ in any respect from yolks of low
albumen quality eggs.
Both the firm and the fluid albumen
fractions of high quality eggs contained
630 L. W. CHARKEY, ELIZABETH DYAR, AND H. S. WILGUS, J R .
more solid matter than did their low
quality counterparts. In high quality
eggs the fluid albumen contained signifi-
cantly more solid matter than did the
firm albumen. It is important to note that
in low quality eggs no such difference
was observed. The distribution of water in
such eggs was at a level throughout the
entire white of the egg. The results re-
ported here confirm those of St. John
(1936), who reported a significantly higher
moisture content in the firm albumen
than in the fluid albumen of selected
"non-watery" eggs. In his "watery"
(low quality) eggs he found a small differ-
ence in the opposite direction, of doubtful
statistical significance. These data may
indicate the breakdown, in low albumen
quality eggs, of a water regulating
mechanism present in eggs of higher
quality.
The distribution of total nitrogen (and
presumably of protein) corresponds to
that of total solids, as appears in Table 1.
This is in conformity with the fact that
the solids content of egg white is very
largely protein. However, in the firm
albumen fraction, whose amount and con-
sistency is the principal criterion of egg
quality, there is more nitrogen contained
in a gram of high quality solids than in a
gram of low quality solids. It seems proba-
ble that the type or composition of the
protein complex varies between high and
low quality eggs, thus accounting for a
greater water-holding capacity in low
quality eggs. The relatively firm, less
hydrated solids of high quality eggs ap-
pear to be richer in content of nitrogen.
Finally an indication has been ob-
tained that the content of one of the spe-
cific protein building units, tryptophane,
is higher in high albumen quality eggs
than in eggs of low quality. The statistical
significance of the difference found in the
fluid albumen comparison is beyond ques-
tion. The firm albumen comparison gave a
smaller difference of less certain statistical
significance, but again in favor of the high
quality eggs. In the yolks no significant
difference was shown.
This variation in apparent tryptophane
content is subject to several possible ex-
planations. First it is conceivable that
there is variation in the tryptophane con-
tent of "specific" proteins. A second ex-
planation, which appears more likely, is
that variation exists in the ratio between
amounts of the several proteins present.
Third, the microbiological assay medium
used in the measurement of tryptophane
may have been deficient in some unrecog-
nized factor stimulatory to L. arabinosus,
and which is provided in higher amounts
by high quality eggs than by low quality
eggs. Fourth, there may have been some
free tryptophane present, in larger amounts
in high quality eggs than in low quality
eggs.
Confirmation of the finding of variable
egg content of tryptophane, and the ex-
planation of this variation, are the sub-
jects of further studies which are in
progress.
CONCLUSIONS
1. Yolks from hens' eggs of high al-
bumen quality do not differ from those of
low quality eggs in content of total solids,
total nitrogen, or 1( —) tryptophane.
2. Both the fluid and the firm albumen
fractions from high quality eggs have
greater contents of total solids than do the
corresponding low quality fractions.
3. In high quality eggs the fluid al-
bumen contains more solid matter per
gram than does the firm albumen. No
such difference was found in low quality
eggs.
4. The nitrogen content of fresh egg
 NUTRIENT CONTENT OF FRESH EGGS
white fractions corresponds roughly to the
content of total solids, but there is more
nitrogen per gram of high quality firm
albumen solids than per gram of low
quality firm albumen solids. In addition
an indication has been obtained that
fluid and firm albumen fractions from
high quality eggs contain higher levels of
l(-)tryptophane than do the corre-
sponding fractions from low quality eggs.
5. It appears likely that the type or
composition of the protein complex
varies between eggs of high and low
albumen quality, especially in the firm
albumen fraction, resulting in a reduced
water holding capacity in high quality
eggs. The solids from such eggs are higher
in nitrogen content than are the solids
from relatively watery, low quality eggs.
ACKNOWLEDGEMENT
We wish to express our appreciation for
the large amount of technical assistance
rendered by Mr. Eldon G. Hill during the
course of this study.
631
REFERENCES
Assoc, of Official Agr. Chem., 1940. Methods of
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Ibid. p. 308.
Balls, A. K. and S. R. Hoover, 1940. Behavior of
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and Eng. Chem. 32: 594-596.
Cheldelin, Vernon H., Margaret A. Eppright, Es-
mond E. Snell and Beverly M. Guirard, 1942.
Enzymatic Liberation of B Vitamins from Plant
and Animal Tissues. Univ. of Texas Publ. 4237:
15-36.
Cole, R. K., 1938. Histological Basis for the Differ-
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Conrad, R. M. and H. M. Scott, 1939. Changes in
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, 1942. Differences between High and Low
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Harte, Robert A., 1945. On the In Vitro Proteolysis
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Sharp, Paul F., 1937. Preservation and Storage of
Hens' Eggs. Food Res. 2:477-498.
Snedecor, George W., 1940. Statistical Methods.
Iowa State College Press. Ames, Iowa.
St. John, J. L., 1936. What Is a "Watery" Egg?
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