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Chemical Method For Determination of Carbon Dioxide Content in Egg Yolk and Egg Albumen
Chemical Method For Determination of Carbon Dioxide Content in Egg Yolk and Egg Albumen
INTRODUCTION
The safety, quality, and shelf life of shell eggs is a
function of carbon dioxide content. A commercial process
was recently developed for rapidly cooling shell eggs
using cryogenic CO2. The benefit of this new process over
existing cooling processes is that CO2 addition during
cryogenic cooling provides additional safety and quality
enhancements. In order for these benefits to be fully developed into a process that can be adopted by the egg
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KEENER ET AL.
FIGURE 1. Equipment used to determine CO2 concentration in albumen and yolk samples.
[1]
[2]
mg of CO2
=
g albumen
[mEq
mg of CO2
mEq
[3]
1
.
of alabumen
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age.5 In addition, an analysis with SAS PROC GLM6 evaluated the statistical equivalence of grouped means based
on sample size and added CO2 solution. Interaction effects
and non-normality concerns were also evaluated using
SAS software.6 Statistical significance and mean grouping
were based on P < 0.01.
[4]
YmEq
Normality
ZmL
[5]
Statistical Analyses
Means and standard errors were obtained for each sample pair using Microsoft Excel Statistical Analysis Pack-
5
Microsoft Excel, Version 97, Microsoft Corp., Redmond, WA
98052-6399.
6
SAS software, Version 6.12, SAS Institute Inc., Cary, NC 27513.
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KEENER ET AL.
TABLE 1. Summary of egg albumen CO2 concentration data1
Mass2 (g)
CO2 added
(mL)
Average CO2
content2 (mg)
Average albumen
CO2 concentration
(mg/g albumen)
0.00
1.99
3.99
6.04
8.05
10.04
0.00
1.97
4.03
6.04
7.99
9.9
0.00
2.12
4.01
5.99
8.02
9.99
0.00
0.00
0.00
0.00
0.00
0.00
2.00
2.00
2.00
2.00
2.00
2.00
4.00
4.00
4.00
4.00
4.00
4.00
0.82
2.57
6.14
9.37
13.53
18.40
1.69
5.12
8.57
11.18
15.06
17.89
3.56
7.27
10.27
15.26
16.79
19.56
...
1.57
1.74
1.68
1.78
1.91
...
1.68
1.78
1.62
1.71
1.65
...
1.83
1.67
1.94
1.64
1.60
0.014
0.014
0.035
0.000
0.063
0.07
0.124
0.063
0.035
0.049
0.077
0.000
0.049
0.077
0.127
0.120
0.036
0.280
0.040
0.401
3.28
0.160
0.601
0.120
0.200
0.481
0.320
1.121
0.280
0.361
1.803
0.200
0.441
0.194a
0.076a
0.016a
0.049a
0.314a
0.232a
0.029a
0.016a
0.067a
0.023a
0.068a
0.089a
0.284a
0.008a
0.023a
Mass2 (g)
CO2 added
(mL)
Average CO2
content2 (mg)
Average yolk
CO2 concentration
(mg/g yolk)
0.00
2.05
4.05
5.97
8.00
9.97
0.00
1.99
4.00
6.01
7.99
9.94
0.00
1.99
4.03
6.02
7.97
10.04
0.00
0.00
0.00
0.00
0.00
0.00
2.00
2.00
2.00
2.00
2.00
2.00
4.00
4.00
4.00
4.00
4.00
4.00
0.76
0.19
0.34
0.50
0.67
0.50
1.89
1.83
2.14
2.77
2.51
2.26
4.10
4.61
4.78
4.35
5.37
5.15
...
0.50
0.11
0.22
0.18
0.13
...
0.23
0.19
0.23
0.14
0.089
...
0.52
0.30
0.13
0.22
0.15
0.028
0.014
0.141
0.035
0.056
0.042
0.077
0.120
0.063
0.000
0.007
0.014
0.014
0.021
0.014
0.440
0.120
0.320
0.160
0.160
0.080
0.120
0.841
0.481
0.080
0.120
0.240
0.200
0.280
1.082
0.401
0.561
0.240
0.066a
0.079a
0.028a
0.019a
0.008a
0.427a
0.123a
0.017a
0.016a
0.024a
0.138a
0.269a
0.066a
0.069a
0.024a
TABLE 3. Average CO2 concentrations determined for egg albumen, egg yolk, and CO2 solution samples
CO2 Concentration1
CO2 Solution (mg/mL)
Albumen + CO2
Yolk + CO2
CO2 Solution
CO2 Solution
Literature2
1.01
1.11
1.23
1.26
...
0.14
0.11a
0.11a
0.08a
Albumen (mg/g)
Yolk (mg/g)
1.66 0.03
...
...
...
1.90
0.09 0.02
...
...
0.00
987
Albumen blank
Yolk blank
Albumen
Yolk
CO2 Content1
(mg)
CO2 Concentration1
(mg/g)
Coefficient of
variation
(%)
0.938 0.033
0.158 0.006
8.64 0.0024
0.456 0.0000132
...
...
1.44 0.0004
0.076 0.0000022
3.5
3.8
0.028
0.0029
eggs were pooled, and five sample blanks, five 6-g yolk
samples, and five 6-g albumen samples were made. These
eggs were then tested using the prescribed method, and
the results are shown in Table 4. From these results, one
can observe that the CO2 concentration in the fresh egg
albumen and egg yolk is 1.44 mg/g and 0.076 mg/g,
respectively. The CV for the albumen and yolk were 0.03
and 0.003%, respectively. These results suggest that using
five 6-g sample averages will allow measurements to be
within 0.03% precision for albumen and 0.003% precision
for yolk. In practice, this method produced CO2 measurements in albumen and yolk with CV of 5% or less for 15
6-g samples. The large increase in sample size and CV
resulted from CO2 content differences between individual eggs.
In conclusion, these results suggest that the chemical
method presented here can be accurately and precisely
used to measure CO2 concentrations in egg albumen and
egg yolk samples. Emphasis should be placed on determining the background CO2 levels because they can
vary considerably with time. In our laboratory, this
method of CO2 measurement with 15 6-g egg albumen
and egg yolk samples has consistently produced CV of
5% or less. This developed method for determining CO2
content also has possibilities in other food systems because it has been shown to work in high protein (egg
albumen) and high lipid (egg yolk) products.
ACKNOWLEDGMENTS
The authors would like to acknowledge the assistance
of Roger Thompson of the USDA Fermentation Labora-
tory (North Carolina State University, Raleigh, North Carolina) who provided details on the method developed
for determining CO2 content in cucumber brines.
REFERENCES
Brey, W. S., 1978. Physical Chemistry and Its Biological Applications. Academic Press Inc., New York, NY.
Cotterill, O. J., F. A. Gardner, F. E. Cunningham, and E. M.
Funk, 1959. Titration curves and turbidity of whole egg
white. Poultry Sci. 38:836842.
Fleming, H. P., R. L. Thompson, and J. L. Etchells, 1974. Determination of carbon dioxide in cucumber brines. Assoc. Off.
Anal. Chem. J. 57:130133.
Heath, J. L., 1977. Chemical and related changes in egg albumen
during storage. Poultry Sci. 56:822828.
Mueller, W. J., 1958. Shell porosity of chicken eggs 1. CO2 loss
and CO2 content of infertile eggs during storage. Poultry Sci.
37:437444.
Official Methods of Analysis, 1995a. AOAC Official Method
940.17Carbon Dioxide in BeerManometric Method. Section 27.1.30. 16th ed. AOAC International, Arlington, VA.
Official Methods of Analysis, 1995b. AOAC Official Method
923.02Carbon Dioxide (Total) in Baking PowdersGasometric Determination. Section 25.1.02. 16th ed. AOAC International, Arlington, VA.
Official Methods of Analysis, 1995c. AOAC Official Method
920.194Carbonate and Bicarbonate in WaterTitrametric
Method. Section 11.1.17. 16th ed. AOAC International, Arlington, VA.
Reinke, R. C., and R. C. Baker, 1966. Relationship between carbon dioxide permeability and bacterial penetration in chicken
egg shell models. Poultry Sci. 45:13271334.