Prevention of the Green-Gray Discoloration in
Cooked Liquid Whole Eggs
PATRICIA W. GOSSETT and ROBERT
~ ABSTRACT
The prevention of :he green-gray discoloration of cooked liquid
whole eggs was stud ed. Acidic chelating agents were added to raw
liquid whole egg samples at pH 8.50. Samples were cooked at 100°C
for 20 mln, held over a steam bath for 60 min, and then measured
for color. Results shew the following approximate optimum concen-
trations will preven. the discoloration at p = 0.05: acetic acid,
0.19%; citric acid, C.17%; NaZEDTA, 0.029%; lactic acid, 0.27%;
malic acid, 0.22%; nlonosodium phosphate, 0.34%; propionic acid,
0.26%; and succinic acid, 0.27%. It was also found that the average
maximum pH attained by raw liquid whole eggs when stored at
3°C for 21 days was 7.93 + 0.19. A taste panel indicated that there
were no significant differences in overall acceptability with the
Na*EDTA and the lnonosodium phosphate treatments. The citric
acid treatment was llot significantly different from the control in
flavor acceptability. Other treatments were judged to be inferior
to the control.
INTRODUCTION
IT HAS BEEN OE SERVED that a green-gray discoloration
may develop in cooked liquid whole eggs. Tinkler and Soar
(1920) noted that the greenish-black color produced on the
surface of hard-coe,ked egg yolks was due to ferrous sulfide
(FeS) formation from the iron in the yolk and the hydro-
gen sulfide produc:ed from heating of the albumen. Baker
et al. (1967) fount i that the greenish-black discoloration on
hard-cooked egg folks was influenced by high cooking
temperature, long cooking time, pH of the yolk, long stor-
age time of eggs bl:fore cooking, and method of cooling the
cooked egg.
Salwin et al. (1953) suggested that the green color in
cooked whole egg or egg yolk formed under conditions of
heat and high pH The color formation was inhibited by
addition of a sm:Jl quantity of the tetrasodium salt of
ethylene diamine :etraacetic acid (Na4EDTA). Kline et al.
(1953) noted that dried whole eggs having a reconstituted
pH in the region 3.6-9.0 exhibited the greening phenome-
non. By adding Na,&EDTA at the level of 0.077% by weight,
the greening was inhibited. Trelease et al. (1952) adjusted
the pH of cooked albumen in canned hard-boiled eggs in
order to preserve the flavor, color, and texture of the heat-
treated eggs. The pH of the albumen was adjusted to be-
tween 5.16 and 6.,1 by soaking or canning the cooked eggs
in edible acids or as:idic buffer systems.
Gravani (1969) found that 0.25% citric acid, 0.04%
lactic acid, 0.07% EDTA, 0.30% sodium acid pyrophos-
phate, and 0.40% monosodium phosphate prevented
greening in cooker liquid whole eggs. Ziegler et al. (1971)
found the green color development was prevented by acid-
ifying eggs to aboi!t pH 6.8-7.1 with an edible acid, prefer-
ably citric acid. Ng (1971) reduced the gray or green dis-
coloration at the yolk-albumen interface of cooked egg
products by treatir g the raw egg yolk with an edible oxidiz-
ing agent such as hydrogen peroxide. Chin and Redfern
Authors Gossett and Baker are affiliated with the Departments of
Poultry & Avian Sciences and Food Science, Rice Hail, Cornell
Univ., Ithaca, NY 141’53.
328-Volume 46 (1981/--JOURNAL OF FOOD SCIENCE
C. BAKER
(1968) used small amounts of soluble phosphorus com-
pounds added to liquid whole egg to prevent color change
from the characteristic orange-yellow to a muddy brown
color.
The goal of this study was to find the optimum concen-
tration of various acidic chelators added to the raw liquid
whole egg that would prevent greening in cooked liquid
whole eggs. It was reasoned that the chelator would tie up
the iron thus making it less available for FeS formation.
The acidic nature of the chelator was needed to inhibit FeS
formation by possibly inhibiting hydrogen sulfide produc-
tion upon heating or by not favoring sulfide ion formation.
A preliminary study was done to determine the maxi-
mum pH that raw liquid whole egg would reach upon stor-
age as liquid whole egg and to see if storage time as shell
eggs prior to breaking affected the maximum pH attained.
EXPERIMENTAL
Eggs
All eggs were from a single strain of White Leghorn hens. Eggs
varied in age from 1-14 days old unless otherwise stated.
Preliminary determinationof maximum pH attained
by raw liquid whoie egg during storage
Shell eggs(0, 3, 6,9, 12, and 15 wk old) that had beenstoredat
3°C were studied. Five eggsper storage period were broken, blended
in a Waring Blendor at low speed for 5 set, strained, and stored in a
plastic container with a snap-on lid. The pH was measured on a
Beckman Expandomatic pH meter daily for 14 days and then every
2 days until a total of 21 days elapsed. Between pH measurements,
the liquid whole egg was stored at 3°C. The entire experiment was
repeated.
Sample preparation
Eggs were broken, blended in a Waring Blendor at low speed for
5 set, and strained. The eggs were adjusted to pH 8.50 with 2N
NaOH to insure geening in the control. Fiftygram samples of
liquid whole egg were weighed out into 150 ml beakers. Treat-
ments consisted of adding different volumes of a 5% solution of
acetic acid, citric acid, disodium ethylene diamine tetraacetic acid
(Na*EDTA), lactic acid, malic acid, monosodium phosphate,
propionic acid, or succinic acid to the liquid whole egg samples.
The mixtures were then stirred and pH measurements were made.
Twelve samples per treatment and at least three treatments per
chelator were prepared.
Cooking and holding procedure:
The samples were cooked without stirring for 20 min in a 100°C
water bath with a sheet of foil fitted loosely over the top of the wa-
ter bath. The samples were then held for 60 min over a steam bath
to approximate steam table treatment.
Color measurement
The cooked egg was removed from each beaker and a sample
was sliced approximately 8 mm from the bottom. The egg slice
was placed in a 60 x 15 mm plastic petri dish and the color mea-
sured on a Hunterlab Color Difference Meter (Model D25-2). The
meter was calibrated with a yellow standardtile which had.color
values of L = 78.2, a = -2.3, and b = 22.5. Duplicate readings of
the same sample varied approximately _+0.2 unit for L, a, and b
values. The average standard deviations for all treatments at the
 DISCOLORATION IN COOKED L lOUID EGGS. . .

optimum concentration level for L, a, and b values were approxi-
mately 1.9, 1.7, and 2.1 units respectively.
The samples were also visually inspected and given a score of
(+) if discoloration were present and (-) if discoloration was absent.
Analysis of data
The proportion of chelator-treated samples that turned green
after cooking was estimated by:
differences between the sample products and the control with re-
spect to flavor and to texture, using a scale of 4 = very different,
3 = moderately different, 2 = slightly different, and 1 = no differ-
ence. The acceptability of the sample products and the control
was rated on a 9 point scale with 9 = excellent, 5 = neutral, and 1 =
poor with respect to flavor, texture, color, and overall acceptability.
The design of the taste panel evaluation was that of a randomized
block.

#of green samplespost-treatment RESULTS&DISCUSSION

P=
#of samplesper treatment
Assuming normal distribution, the p values can be converted to
Z values, the standard normal variate, in order to linearize the data.
The Z value corresponds to the area under the normal probability
curve from -m to Z. The Z values were plotted against the concen-
tration of acidic chelator used and a linear regression was performed
on the data. A level of 5% greening was arbitrarily set as being ac-
ceptable with p = 0.05 and Z = -1.64. At this Z level, the concen-
tration of chelator needed to produce greening at the 5% level could
be calculated.
For example, if 0.183% w/w of malic acid was used in a certain
treatment, the proportion of samples remaining green post-treat-
ment was 2/3 or p = 0.667. The corresponding Z value is 0.431.
If 0.192% w/w of malic acid was used in another treatment, the pro-
portion of samples remaining green was l/4 or p = 0.250, with a Z
= -0.675. In Figure 1, the fraction of green samplespost-treatment
in terms of Z values versus the concentration of malic acid is shown.
Note that at Z = -1.64 which corresponds to 5% greening, the con-
centration predicted is 0.22%. This was defined as the optimum con-
centration needed to prevent greening.
The final result is expressed in terms of weight %:
ml of chelator soln % liquid whole egg population should be equal to or less than
weight % = x -x 100
5O.Og+ ml of chelator soln 100 ml
Assume that 1 ml of chelator solution is approximately 1 g.
At the predicted optimum concentration for each chelator, 12
samples were prepared and measured for pH and color as described
previously.
Preliminary determination of maximum pH attained
by raw liquid egg during storage
Figure 2 shows the average pH per treatment plotted
against the number of days stored as liquid egg at 3’C.
Note that the eggs reached a maximum pH regardless of
treatment; zero wk old shell eggs (fresh) reached their
maximum slower than the other eggs that had been stored
longer as shell eggs. After 5 days of storage, the pH of fresh
eggs had risen from approximately 7.1 to 7.5. The reason
for the observed decline in pH is not known at this time.
The average maximum pH attained by raw liquid -whole
egg from two trials per treatment during 21 days of storage
at 3°C as liquid whole egg is shown in Table 1. The average
maximum pH attained does not differ significantly from
treatment to treatment as determined by a least significant
difference test. Therefore, regardless of the age of the shell
eggs, the liquid whole egg in this experiment reached an
average maximum pH of approximately 7.93 * 0.19. The
maximum pH that could be reached by 99.74% of the
the mean pH value f 3 standard deviations or 7.93 + 3
(0.19) = 8.50.
For the purpose of this experiment, the initial pH to
which the raw liquid whole eggs should be adjusted should

Taste panel analysis of the cooked egg products 8.1

The cooked egg products with the added chelators at the opti-
mum concentrations were evaluated by a 15-member taste panel 8.0
against a control egg sample with no chelator added. One day old
eggs were used as the control. The panelists were asked to compare
7.9

7.8

7.7

I 7.6
*
7.5 AS SHELL EGGS: .-
O---O 15 WEEKS -a
7.4 4: o--o 12 WEEKS
i s-4 9 WEEKS
E
7.3 1;: ~....a 6 WEEKS
:-- n 3 WEEKS
7.2 i *-..+ 0 WEEKS
j

7.1

7.0

WEIGHT % MALIC ACID DAYS STORED

AS LIQUID EGGS
I-Plot of the fraction of green samples post-treatment in
terms of i’ values versus the concentration of malic acid. Note that Fig. 2-Plot of the pH of liquid whole egg from shell eggs of various
at Z = - 1.64, tie predicted concentration is 0.2%. ages versus the number of days stored as liquid whole egg at 3°C.

Volume 46 /1981)-JOURNAL OF FOOD SCIENCE-329

Table l-Average maximum pH attained by raw liquid whole egg be a pH level at which greening is exhibited but still ap-
during 2 1 days of sto *age as liquid whole egg at 3” C proximate a realistic environment in which eggsexist. Kline
et al. (1953) noted greening took place at pH 8.6-9.0. Since
Wk of storage Average the maximum pH of liquid eggs was calculated to be 8.50,
as shell eggs max pH raising the pH to 9.0 may not approximate a realistic en-
0 7.88 f 0.06
vironment for the egg. A pH of 8.50 rather than 7.93 was
3 7.95 f 0.11 chosen as the initial pH to which the eggs were to be ad-
6 7.93 f 0.04 justed because it represented an uppermost alkaline condi-
9 7.90 + 0.01 tion which insured exhibition of the greening phenomenon.
12 7.99 f 0.13
15 7.94 f 0.05 Determination of optimum concentrations
Grand Average = 7.93 + 0.19 of acidic chelators
The concentrations of the various chelators tested were
chosen such that at those concentrations, a fraction of the
samples remained green post-treatment. Thus, p, the esti-
mate of the proportion of greening, could be calculated. By
extrapolation one could calculate the concentration needed
Table P-For each cfelator shown, the concentrations tested along
in order to have only 5% of the samples turning green with
with the resulting pti of the raw liquid whole egg, the proportion p = 0.05. In Table 2, the concentrations for each chelator
of greening at that concentration, the corresponding Z value, and tested, the pH at each concentration, the proportion that
thepredicted:koncenirationsatp =0,05arepresented turned green at each concentration, and the corresponding
Z values are shown. Note that as the concentration of
Core
chelator used is increased, the proportion of samples that
Chelator WIVI PH’ P z turned green post-treatment decreased.
Table 3 shows the conditions for prevention of the
Acetic acid 0.166 6.69 .t 0.01 0.500 0.000 green-gray discoloration in liquid egg at p = 0.05. The L,
0.17 6.58 + 0.03 0.167 -0.966 a, and b values for the cooked egg products for each chel-
0.19 6.46 f 0.04 0.083 -1.383 ator were shown to be significantly different at the 5%
0.21 6.35 f 0.03 0.001 -3.090 level from the control at pH 8.5 as determined by a least
0.19’ 6.42 f 0.07’ 0.050 -1.640
significant difference test.
Citric acid 0.08 7.42 c 0.03 0.999 3.090
The optimum concentration predicted from this study
0.10 7.26 t 0.04 0.667 0.43 1 for citric acid (0.17%) is less than that found by Gravani
0.1 1 7.27 k 0.05 0.917 1.383 (0.25%) and for monosodium phosphate (0.34%) is less
0.14 7.05 k 0.06 0.667 0.431 than that found by Gravani (0.40%) and within the range
0.16 6.91 f 0.03 0.083 -1.383 found by Chin and Redfern (1968) (0.3-0.7%). A possible
O.lJa 6.83 f 0.03’ 0.050 -1.640 reason why the values from this study are lower than values
determined by Gravani is that the pH of the control samples
Na*EDTA 0.01; 8.36 f 0.03 0.750 0.675 of this study was 8.50 whereas those of Gravani’s were pH
0.013 8.33 + 0.02 0.917 1.383
0.02 2 8.36 f 0.04 0.833 0.966
9.1.
0.021 8.38 f 0.02 0.083 -1.383 Taste panel analysis of the cooked egg products
0.02 3a 8.40 f 0.01’ 0.050 -1.640
Statistical analyses of the taste panel data consisted of
Lactic acid 0.14 7.13 f 0.02 0.999 3.090 performing an analysis of variance on the control and the
0.17 6.76 f 0.04 0.500 0.000 eight treatments for flavor difference, texture difference,
0.19 6.66 * 0.07 0.333 -0.431 flavor acceptability, texture acceptability, color acceptabil-
0.21 6.65 + 0.05 0.500 0.000
ity, and overall acceptability. A one-tailed Dunnett’s test
0.23 6.60 f 0.02 0.583 0.210
0.27’ 5.74 f 0.02’ 0.050 -1.640
was then performed at the 5% and 1% significance level by
comparing each treatment mean with that of the control
Malic acid 0.18 6.76 f 0.04 0.917 1.383 (Table 4).
0.19 6.60 f 0.01 0.667 0.431 Table J-Conditions for prevention of green-gray discoloration in
0.21 6.51 i 0.02 0.250 -0.675 liquid whole eggs
0.22s 6.53 f 0.04’ 0.050 -1.640
Color of
Monosodium 0.19 7.46 zi 0.05 0.999 3.090 pH of liquid Cone cooked producta
phosphate 0.24 7.32 + 0.03 0.833 0.966
Chelator egg & chelator wlw L a b
0.28 7.19 * 0.04 0.667 0.431
0.34” 7.08 t 0.02e 0.050 -1.640 Acetic acid 6.42 r 0.07 0.19% 78.8 -1.0 33.0
Citric acid 6.83 f 0.03 0.17 78.9 -1 .o 32.9
Propionic acid 0.22 6.57 f 0.04 0.999 3.090 Na2EDTA 8.40 + 0.01 0.029 72.5 0.3 27.4
0.24 6.58 + 0.12 0.500 0.000 Lactic acid 5.74 t 0.02 0.27 79.7 -1.8 31 .l
0.25 6.53 k 0.06 0.417 -0.210 Malic acid 6.53 f 0.04 0.22 73.3 -1 .J 26.2
0.26s 6.47 + 0.03’ 0.050 -1.640 Monosodium 7.08 f 0.02 0.34 80.0 -2.8 32.3
phosphate
Succinic acid 0.16 6.76 i 0.03 0.667 0.431 Propionic acid 6.47 f. 0.03 0.26 74.4 -1.5 28.6
0.19 6.54 f 0.04 0.500 0.000 Succinic acid 6.15 t 0.02 0.27 74.8 0.6 28.1
0.21 6.42 r 0.03 0.250 -0.675
0.2J3 6.15 + 0.06c 0.050 -1.640 Control 8.50 + 0.02 0 47.0 7.6 3.0
Controfb 7.07 f 0.12 0 62.0 1.4 13.9
a Predicted concentration that gives greening at P = 0.05; found by Controlc 7.07 f 0.12 0 76.7 -0.2 30.6
calculation
b Each pH listed is avcwaged frOm 12 samples. a Measured on Hunterlab Color Difference Meter
’ Found experiment;llly at the predicted concentration that gives b Fresh egg sample, cooked 20 min at 100°C
greening at p = 0.05 ’ Fresh egg sample, cooked 10 min at 100°C

330-Volume 46 (i’981)-JOURNAL OF FOOD SCIENCE

 DISCOLORATION IN COOKED LIQUID EGGS. . .

Table 4-Results of taste panel evaluation: one-tailed Dunnett’s tests

Mean difference Mean acceptability

Treatment Flavor Texture Flavor Texture Color Overall

Control 1 .o 1 .o 7.4 7.1 6.4 7.3

Acetic acid 2.9”” 3.4** 4.9** 4.5** 6.9 4.7**
Citric acid 1.6” 2.1** 6.5 5.9” 7.2 6.2”
Na2EDTA 2.1 l * 1.4 6.2’ 6.2 6.4 6.3
Lactic acid 2.8** 3.1” 4.5** 4.4** 7.7** 4.3””
Malic acid 1.9”” 2.2”” 6.0”” 5.9” 7.4* 5.7”
Monosodium phosphate 1.7’ 1.7** 6.2* 6.5 7.7’1 6.5
Propionic acid 2.5”” 2.3** 5.7** 5.8* 7.3* 5.3””
Succinic acid 3.9** 2.8”’ 1.41” 4.5** 7.4” 1.5”’
l Significantly different at the 5% level
**Significantly different at the 1% level

The results from the Dunnett’s test indicated: SUMMARY

Flavor difference-All the added chelators were judged (1) The maximum pH attained by liquid eggs from shell
as being different in flavor from the control at the 1% level eggs of various ages averaged around 7.93 2 0.19 during 2 1
except the citric acid and monosodium phosphate treat- days of storage as liquid whole egg at 3’C. 99.74% of the
ments which differed at the 5% level. population of liquid whole eggs should have a maximum
Texture difference-The Naa EDTA treatment did not pH of 8.50 or less when stored as liquid whole eggsat 3°C.
differ significantly from the control in texture. All other (2) The optimum concentrations of the following acidic
chelators were found to be different at the 1% level. chelators needed to prevent greening with p = 0.05 were
Flavor acceptability-The citric acid treatment was not found to be: acetic acid, 0.19%; citric acid, 0.17%; Naz-
significantly different in flavor acceptability from the EDTA, 0.029%; lactic acid, 0.27%; malic acid, 0.22%;
control. All other treatments differed from the control at monosodium phosphate, 0.34%; propionic acid, 0.26%;
the 1% level except the Na,EDTA and monosodium phos- and succinic acid, 0.27%.
phate treatments which differed at the 5% level. (3) The Naz EDTA (0.029%) and the monosodium phos-
Texture acceptability-The NazEDTA and monosodium phate (0.34%) treatments were rated as being as acceptable
phosphate treatments were not significantly different from (overall) as the control sample. The citric acid (0.17%)
the control with respect to texture acceptability. All other treatment was evaluated as being as acceptable as the con-
treatments were judged as being significantly less acceptable trol sample with respect to flavor.
than the control at the 5% level except the acetic, lactic,
and succinic acid treatments, which were different at the REFERENCES
1% level.
Color acceptability-All treatments were found to be as Baker, R.C.. Darfler, J., and Lifshitz. A. 1967. Factors affecting
the discoloration of hard-cooked egg yolks. Poultry Sci. 46: 664.
acceptable as the control with the malic, propionic, and Chin, R.G.L. and Redfern, S. 1968. Egg compositions containing
succinic acid treatments being significantly more acceptable soluble phosphorus compounds effective to impart fresh egg color.
U.S. Patent 3, 383.221.
at the 5% level, and the lactic acid and monosodium phos- Gravani, R.B. 1969. The formation and prevention of a greenish-
phate treatments being more acceptable at the 1% level. black discoloration in cooked liquid eggs. MS. thesis, Cornell
University, Ithaca, N.Y.
Overall acceptability-The NaaEDTA and monosodium Kline, L., Sonoda, T.T., and Hanson, H.L. 1953. Relative chemical,
phosphate treatments were not significantly different from functional, and organoleptic stabilities of acidified and glucose-
free whole egg powders. Food Technol. 7(11): 456.
control in overall acceptability. The citric acid treatment Ng. W. 1971. Method of improving color of cooked egg products.
was significantly less acceptable at the 5% level while all U.S. Patent 3, 598,612.
other treatments were determined to be less acceptable at Salwin, H., Bloch, I.. and Mitchell, J.H. Jr. 1953. Dehydrated sta-
bilized egg. Importance and determination of PH. Food Technol.
the 1% level. 7(11): 447.
In spite of the harsh condition under which the eggs Tinkler, C.K. and Soar, M.C. 1920. The formation of ferrous sul-
phide in eggs during cooking. Biochemical J. 14: 414.
were tested, that is, fresh eggs of about pH 7.1 with added Trelease. R.D.. Sampson. G.O.. and Alstrand, D.V. 1952. Canning
acidic chelators designed for eggs at pH 8.5, the taste panel of hard-boiled eggs. U.S. Patent 2, 593,223.
evaluation indicated two or three possible chelators that Ziegler, H.F. Jr., Seeley, R.D.. and Holland. R.L. 1971. Frozen egg
mixture. U.S. Patent 3,565,638.
were acceptable. Na, EDTA and monosodium phosphate MS received 6/5/80: revised S/30/80; accepted g/5/80.
prevented discoloration and were as acceptable as the con-
trol when added to liquid eggs. The citric acid treatment
was found to be as acceptable as the control with respect to
flavor.

Volume 46 (1981)-JOURNAL OF FOOD SCIENCE-331