In conjunction with environmental pollution studies, it often is necessary to analyze field-collected eggs that are partially decomposed. The effect of putrefaction upon residue analysis, however, is not known and the experiment reported here was designed to determine if recoveries of DDT, DDE, and DDD differed between fresh and addled eggs and also if putrefaction causes appreciable degradation of DDT

Mulhern, Bernard M.

Reichel, W. L.

English

UNL | Libraries

University of Nebraska - Lincoln
DigitalCommons@University of Nebraska - Lincoln
US Fish & Wildlife Publications US Fish & Wildlife Service
1970
The Effect of Putrefaction of Eggs Upon Residue
Analysis of DDT and Metabolites
Bernard M. Mulhern
Patuxent Wildlife Research Center
W. L. Reichel
Patuxent Wildlife Research Center
Follow this and additional works at: https://digitalcommons.unl.edu/usfwspubs
This Article is brought to you for free and open access by the US Fish & Wildlife Service at DigitalCommons@University of Nebraska - Lincoln. It has
been accepted for inclusion in US Fish & Wildlife Publications by an authorized administrator of DigitalCommons@University of Nebraska - Lincoln.
Mulhern, Bernard M. and Reichel, W. L., "The Effect of Putrefaction of Eggs Upon Residue Analysis of DDT and Metabolites" (1970).
US Fish & Wildlife Publications. 525.
https://digitalcommons.unl.edu/usfwspubs/525
The Effect of Putrefaction of Eggs Upon Residue 
Analysis of DDT and Metabolites 
by BERNARD M. MULHERN and W. L. REICHEL 
Bueau of Sport Fisheries and Wildlife, Patuxent Wildlife Research Center 
Laurel, Maryland 
In conjunction with environmental pollution studies, it often 
is necessary to analyze field-collected eggs that are partially 
decomposed. The effect of putrefaction upon residue analysis, 
however, is not known and the experiment reported here was 
designed to determine if recoveries of DDT, DDE, and DDD differed 
between fresh and addled eggs and also if putrefaction causes 
appreciable degradation of DDT. 
Procedure 
Thirty-four fresh chicken eggs were randomly selected from 
"large" size eggs purchased on the open market. The samples, 
except for controls, were fortified with p,p'-DDT, p,p'-DDE, and 
p,p'-DDD according to the plan outlined in Table 1. 
The eggs in group C, D, E, and F were fortified soon after 
purchase. Fortification was accomplished by syringe injection 
of the standard solution (~g/~l) through the shell and well into 
the egg. The hole produced in the shell was covered with hot 
paraffin. The eggs in group C and D were frozen immediately. 
Those in groups E and F were allowed to rot in an outdoor open 
shed, and the rate of decomposition was checked periodically by 
noting the condition of extra eggs. A period of 4 weeks, during 
the month of September, was required to obtain satisfactorily 
"rotten" eggs, which then were frozen. 
One additional group of eggs (group A) was frozen fresh 
without injection and one group (group B) was rotted without 
injection and then frozen. These samples were fortified when 
they were opened later at the time of analysis. Two eggs from 
each of the fresh and rotted group were not fortified and served 
as background controls. 
In addition, five randomly selected fresh eggs from hens fed 
25 ppm DDT were frozen fresh and five were allowed to rot with 
the other sets. 
All specimens were stored in a freezer at -27oC for 3 years 
before they were individually analyzed. 
222 
Bulletin of Environmental Contamination & Toxieology, 
Vol. 5, No.3, 1970, published by Springer.Verlag New York Inc. 
Analysis 
Each specimen, including shell, was mixed with anhydrous 
sodium sulfate, extracted in a soxhlet, and cleaned up by 
acetonitrile partitioning and florisil column chromatography 
according to the method of Reichel and Addy (1). The pesticides 
in the clean extract were analyzed by electron capture gas 
chromatography on a 3% OV-17 column and residues were confirmed 
on 3% XE-60 column as described by Reichel et al. (2). 
Results and Discussion 
The results of the analysis of the fortified eggs are shown 
in Table 1 as averages of the five individual determinations in 
each group. The average background level of the control eggs 
was only 1.6 micrograms of DDE and 1.3 micrograms of DDT, and 
was not substracted from the values obtained for the experimental 
eggs. 
A statistical F-test analysis of the data showed no significant 
difference (P<O.05) in the recovery of pesticides from fresh and 
rotten eggs except that the percentage recovery of DDD from rotted 
eggs fortified with 200 micrograms was greater than from any other 
group. The higher values of DDE and DDD in the decomposed eggs 
(group E and F) suggest that there was some degradation of DDT. 
The results of analysis of eggs from hens fed DDT is shown in 
Table 2. Since these results are reported on a ppm basis, it was 
necessary to adjust the wet weight of the eggs to compensate for 
the loss of moisture. The fresh weight was estimated from the egg 
shell dimensions (length x breadth) using the formula suggested by 
Romanoff (3). A statistical F-test of the data showed no significant 
difference between the rotten and fresh samples in the levels of DDE 
or DDT. It is interesting to note that the fresh eggs lost more 
moisture during freezer storage ~n did the rotten eggs. 
These results show that putrefaction of eggs did not have an 
appreciable effect on the recovery of DDT and metabolites nor cause 
conspicuous degradation of DDT. 
Acknowledgments 
The authors appreciate and thank our colleagues for assistance 
in this study; namely, Dr. Lucille Stickel for help in setting up 
the study and preparation of eggs for analysis, Mr. William Stickel 
for providing eggs from DDT-dosed hens, and Mr. Robert Heath for 
assistance in statistical testing of the results. 
~ 
TABLE 1 
Percentage Recovery of Pesticides From Fresh and Rotten Eggs 
Group Condition of No. IJ.g of Each Time of DDE DDD DDT 
Egg Before of Insecticide Insecticide 
Freezing Eggs DDT. DDE. DDD Addition Mean Standard Mean Standard Mean Standard 
Added/Egg Error Error Error 
A Fresh 5 100 Before 102 6.2 86 1.3 84 .5.0 
Extraction 
B Rotten 5 100 Before 93 6.2 82 3.7 86 3.4 
Extraction 
-----------------------------------------------------------~-----------------------------------------------
C Fresh 5 100 
E Rotten 5 100 
D Fresh 5 200 
F Rotten 5 200 
*Significant difference at 5% level 
Before 
Freezing 
Before 
Rotting 
Before 
Freezing 
Before 
Rotting 
105 
113 
117 
120 
5.7 92 6.0 88 7.3 
9.8 85 5.7 83 1.6 
10.5 88 3.6 82 5.2 
3.3 100* 8.2 80 8.2 
TABLE 2 
Residues in Eggs From Hens Fed p,p'-DDT 
Group Wet Corrected Residues, ppm Corrected Wet Weight 
Weight Wet* 
Weight p,p'-DDE p,p'-DDD p,p'-DDT 
Fresh 29.40 52.03 5.21 11.41 
38.19 52.20 2.62 <0.05 9.95 
36.18 53.06 6.00 <0.05 10.98 
39.82 53.06 3.15 <0.05 6.79 
38.41 54.72 5.27 <0.05 9.35 
Mean 4.45 9.70 
Rotten 46.62 51.34 2.73 <0.05 6.31 
48.42 52.92 2.61 <0.05 6.81 
47.80 50.49 5.33 <0.05 10.04 
50.96 53.82 6.24 <0.05 11. 77 
43.67 48.11 2.78 <0.05 6.89 
Mean 3.94 8.36 
*Corrected for loss of weight as described in text 
References 
1. W. L. Reichel and C. E. Addy, Bull. Environ. Contam. and Toxico1., 
l, 174 (1968). 
2. W. L. Reichel, T. G. Lamont, E. Cromartie, and L. N. Locke, ibid, 
~, 24 (1969). 
3. A. L. Romanoff and A. J. Romanoff, The Avian Egg, p. 108 (1963), 
John Wiley and Sons, New York. 
225 


The Effect of Putrefaction of Eggs Upon Residue Analysis of DDT and Metabolites

DigitalCommons@University of Nebraska

University of Nebraska - LincolnDigitalCommons@University of Nebraska - LincolnUS Fish & Wildlife Publications US Fish & Wildlife Service1970The Effect of Putrefaction of Eggs Upon ResidueAnalysis of DDT and MetabolitesBernard M. MulhernPatuxent Wildlife Research CenterW. L. ReichelPatuxent Wildlife Research CenterFollow this and additional works at: https://digitalcommons.unl.edu/usfwspubsThis Article is brought to you for free and open access by the US Fish & Wildlife Service at DigitalCommons@University of Nebraska - Lincoln. It hasbeen accepted for inclusion in US Fish & Wildlife Publications by an authorized administrator of DigitalCommons@University of Nebraska - Lincoln.Mulhern, Bernard M. and Reichel, W. L., "The Effect of Putrefaction of Eggs Upon Residue Analysis of DDT and Metabolites" (1970).US Fish & Wildlife Publications. 525.https://digitalcommons.unl.edu/usfwspubs/525The Effect of Putrefaction of Eggs Upon Residue Analysis of DDT and Metabolites by BERNARD M. MULHERN and W. L. REICHEL Bueau of Sport Fisheries and Wildlife, Patuxent Wildlife Research Center Laurel, Maryland In conjunction with environmental pollution studies, it often is necessary to analyze field-collected eggs that are partially decomposed. The effect of putrefaction upon residue analysis, however, is not known and the experiment reported here was designed to determine if recoveries of DDT, DDE, and DDD differed between fresh and addled eggs and also if putrefaction causes appreciable degradation of DDT. Procedure Thirty-four fresh chicken eggs were randomly selected from "large" size eggs purchased on the open market. The samples, except for controls, were fortified with p,p'-DDT, p,p'-DDE, and p,p'-DDD according to the plan outlined in Table 1. The eggs in group C, D, E, and F were fortified soon after purchase. Fortification was accomplished by syringe injection of the standard solution (~g/~l) through the shell and well into the egg. The hole produced in the shell was covered with hot paraffin. The eggs in group C and D were frozen immediately. Those in groups E and F were allowed to rot in an outdoor open shed, and the rate of decomposition was checked periodically by noting the condition of extra eggs. A period of 4 weeks, during the month of September, was required to obtain satisfactorily "rotten" eggs, which then were frozen. One additional group of eggs (group A) was frozen fresh without injection and one group (group B) was rotted without injection and then frozen. These samples were fortified when they were opened later at the time of analysis. Two eggs from each of the fresh and rotted group were not fortified and served as background controls. In addition, five randomly selected fresh eggs from hens fed 25 ppm DDT were frozen fresh and five were allowed to rot with the other sets. All specimens were stored in a freezer at -27oC for 3 years before they were individually analyzed. 222 Bulletin of Environmental Contamination & Toxieology, Vol. 5, No.3, 1970, published by Springer.Verlag New York Inc. Analysis Each specimen, including shell, was mixed with anhydrous sodium sulfate, extracted in a soxhlet, and cleaned up by acetonitrile partitioning and florisil column chromatography according to the method of Reichel and Addy (1). The pesticides in the clean extract were analyzed by electron capture gas chromatography on a 3% OV-17 column and residues were confirmed on 3% XE-60 column as described by Reichel et al. (2). Results and Discussion The results of the analysis of the fortified eggs are shown in Table 1 as averages of the five individual determinations in each group. The average background level of the control eggs was only 1.6 micrograms of DDE and 1.3 micrograms of DDT, and was not substracted from the values obtained for the experimental eggs. A statistical F-test analysis of the data showed no significant difference (P<O.05) in the recovery of pesticides from fresh and rotten eggs except that the percentage recovery of DDD from rotted eggs fortified with 200 micrograms was greater than from any other group. The higher values of DDE and DDD in the decomposed eggs (group E and F) suggest that there was some degradation of DDT. The results of analysis of eggs from hens fed DDT is shown in Table 2. Since these results are reported on a ppm basis, it was necessary to adjust the wet weight of the eggs to compensate for the loss of moisture. The fresh weight was estimated from the egg shell dimensions (length x breadth) using the formula suggested by Romanoff (3). A statistical F-test of the data showed no significant difference between the rotten and fresh samples in the levels of DDE or DDT. It is interesting to note that the fresh eggs lost more moisture during freezer storage ~n did the rotten eggs. These results show that putrefaction of eggs did not have an appreciable effect on the recovery of DDT and metabolites nor cause conspicuous degradation of DDT. Acknowledgments The authors appreciate and thank our colleagues for assistance in this study; namely, Dr. Lucille Stickel for help in setting up the study and preparation of eggs for analysis, Mr. William Stickel for providing eggs from DDT-dosed hens, and Mr. Robert Heath for assistance in statistical testing of the results. ~ TABLE 1 Percentage Recovery of Pesticides From Fresh and Rotten Eggs Group Condition of No. IJ.g of Each Time of DDE DDD DDT Egg Before of Insecticide Insecticide Freezing Eggs DDT. DDE. DDD Addition Mean Standard Mean Standard Mean Standard Added/Egg Error Error Error A Fresh 5 100 Before 102 6.2 86 1.3 84 .5.0 Extraction B Rotten 5 100 Before 93 6.2 82 3.7 86 3.4 Extraction -----------------------------------------------------------~-----------------------------------------------C Fresh 5 100 E Rotten 5 100 D Fresh 5 200 F Rotten 5 200 *Significant difference at 5% level Before Freezing Before Rotting Before Freezing Before Rotting 105 113 117 120 5.7 92 6.0 88 7.3 9.8 85 5.7 83 1.6 10.5 88 3.6 82 5.2 3.3 100* 8.2 80 8.2 TABLE 2 Residues in Eggs From Hens Fed p,p'-DDT Group Wet Corrected Residues, ppm Corrected Wet Weight Weight Wet* Weight p,p'-DDE p,p'-DDD p,p'-DDT Fresh 29.40 52.03 5.21 11.41 38.19 52.20 2.62 <0.05 9.95 36.18 53.06 6.00 <0.05 10.98 39.82 53.06 3.15 <0.05 6.79 38.41 54.72 5.27 <0.05 9.35 Mean 4.45 9.70 Rotten 46.62 51.34 2.73 <0.05 6.31 48.42 52.92 2.61 <0.05 6.81 47.80 50.49 5.33 <0.05 10.04 50.96 53.82 6.24 <0.05 11. 77 43.67 48.11 2.78 <0.05 6.89 Mean 3.94 8.36 *Corrected for loss of weight as described in text References 1. W. L. Reichel and C. E. Addy, Bull. Environ. Contam. and Toxico1., l, 174 (1968). 2. W. L. Reichel, T. G. Lamont, E. Cromartie, and L. N. Locke, ibid, ~, 24 (1969). 3. A. L. Romanoff and A. J. Romanoff, The Avian Egg, p. 108 (1963), John Wiley and Sons, New York. 225 

https://digitalcommons.unl.edu/cgi/viewcontent.cgi?article=1530&context=usfwspubs

The Effect of Putrefaction of Eggs Upon Residue Analysis of DDT and Metabolites

Abstract

Similar works

Full text

Available Versions

UNL | Libraries

DigitalCommons@University of Nebraska