Aldrin and Dieldrin Residues in Human Serum Collected from Delhi Amit
1 Nair,
’Department Agricultural
P.
2 Dureja
&
M. K. K.
Fat, Milk and Blood
1 Pillai
Division of of Zoology, University of Delhi, and 2 Research Institute, New Delhi - 110012, India.
Agricultural Chemicals,
Indian
1 Aldrin and dieldrin residues were monitored in the fat, breast milk and blood serum from female residents of Delhi. 2 The average aldrin and dieldrin contents were 0.048 and 0.099 ppb in adipose tissue, 0.003 and 0.060 ppb in breast milk and 0.004 and 0.002 ppb in blood serum, respectively. 3 The older donors contained higher levels of aldrin and dieldrin in their adipose tissue. 4 Primagravidae contained more of these chemicals in their breast milk. 5 A positive correlation was observed between the aldrin concentration in adipose tissue and breast milk, and that in adipose tissue and blood serum. Similarly, a significant correlation was found between dieldrin in adipose tissue and blood serum. 6 The levels of aldrin and dieldrin were low in samples from residents of Delhi when compared to those in developed countries. Introduction The extensive use of toxic organochlorine pesticides is causing some concern. Howell’I first reported the storage of DDT in human fat. Since then, other organochlorine pesticides such as lindane, aldrin, dieldrin, heptachlor and its epoxide have been found in the breast milk, blood and adipose tissue of humans from the USA,2 Canada,; 3 Europe and Australia.~ Although there has been increased concern about the pesticide burden of the human body, only a few reports pertaining to the levels of DDT, HCH and HCB are available from India.6-1&dquo; Furthermore, little has been published about the levels of aldrin and dieldrin in human fat, blood and breast milk; only dieldrin levels in placental tissue extracts and breast milk have been cited.&dquo; This communication reports on the aldrin and dieldrin residues found in the fat, blood and breast milk of the women of Delhi.
Methods of human adipose tissue, breast milk and blood were collected from Irwin Hospital, Delhi during the 1989-90. All donors were females belonging to a lower socio-economic class and residing in those parts of Delhi exposed
Samples
to severe automobile
pollution. None of them employed and all were vegetarians. The donors of adipose tissue were between 18-42
were
years of age, while those who donated blood and breast milk were between 20-34 years. All donors had either undergone their first or second delivery. None of the donors had a history of previous abortions, or any serious recent illness. However, all of them had suffered from typhoid and jaundice in the past. Blood and breast milk samples were taken from donors with normal and healthy babies whose birthweight was over 2800 g. Adipose tissue samples were obtained from patients undergoing abdominal surgery. Breast milk samples were manually expressed from lactating mothers only after the infant was nursed (hind milk). The blood samples which were drawn from the median cubital vein of the donors, were immediately centrifuged (2500 rpm, 10 min) to obtain blood serum. In general, the Environmental Protection Agency (EPA) protocol was used for the analysis of aldrin and dieldrin.1 2-14 The adipose tissue samples (5 g) were macerated with sand and anhydrous sodium sulphate and the fat was extracted with repetitive extractions with petroleum ether and acetonitrile. 12Breast milk samples (5 ml) were shell-freezed in liquid nitrogen, lyophilized and Soxhlet extracted with n-hexane
Downloaded from het.sagepub.com at Bobst Library, New York University on May 8, 2015
44
..
,
for 6 h.’3 Blood serum samples (2 ml) were extracted with 6 ml n-hexane for 2 h in a slow speed rotatory mixer and then concentrated to 0.5 ml in a stream of nitrogen. 12 The extracted samples of adipose tissue and breast milk were concentrated in vacuo and cleaned over an alumina column with 60 ml n-hexane.’4 Residues of aldrin and dieldrin were qualitatively and quantitatively determined using a Hewlett Packard model 5890A gas chromatograph equipped with a 63 Ni electron capture detector and coupled with an integrator. A megabore capillary column 0.53 mm i.d and 10 m long, packed with 3% OV-17 was used. The operating conditions were as follows: the column temperature was initially programmed at 200°C for 3 min and was then increased at the rate of 10°C min-’ up to a final temperature of 250°C. The detector and injector were maintained at 300°C. Nitrogen with a flow rate of 50 ml min-’ was used as the carrier gas. The analytical standards of aldrin and dieldrin were obtained from the EPA. The residues of aldrin and dieldrin were further identified by using gas chromatography/ mass spectrometry (Joel model JMS DX-300).
Results The percentage recoveries of aldrin and dieldrin from spiked samples of adipose tissue, breast milk and blood samples are shown in Table 1. The average recovery was found to be more than 85%. However, the data presented in Table 2 have not been corrected for recovery. The residues of aldrin and dieldrin in human adipose tissue, breast milk and blood serum are given in Table 2. There was more aldrin and dieldrin in fat than in either breast milk or blood
Table 1 Percent recovery of aldrin and dieldrin from spiked human samples.
In addition, levels of dieldrin were twice that of aldrin. However, there was very little variation in the amounts of aldrin and dieldrin in the blood serum. No significant difference in the levels of aldrin could be observed between breast milk and blood serum. Breast milk had a 30 times higher dieldrin concentration than the blood serum.
serum.
’
Discussion The present survey indicates a considerably lower human burden of aldrin and dieldrin in women from Delhi than that reported from other countries.4.5.15,17 As with an earlier report from Australia, human fat contained more dieldrin than aldrin. Older donors contained higher amounts of chemical than younger ones. In breast milk, the mean levels of aldrin and dieldrin were found to be 0.003 ppb and 0.060 ppb, respectively, indicating bioconcentration of dieldrin in the breast. However, the levels of aldrin and dieldrin in milk were much lower than those reported for samples from Denmark or Australia. In the present study, it was also noticed that the levels of these chemicals in breast milk were dependent on the number of parturations. Women who had been delivered for the first time had more aldrin and dieldrin in their breast milk than those who had undergone their second delivery. In addition, older women with their first child had more of these pesticides in their bodies than younger ones. Unlike fat and milk, blood serum did not show any significant difference in aldrin and dieldrin levels; in addition levels in blood were much lower than those reported for samples from the USA 2,18 or
Canada.3
Although the use of dieldrin has been banned in India since 1975, its levels in adipose tissue and breast milk have been surprisingly higher than those of aldrin. Hence, whatever dieldrin has been detected may not actually be environmental in its origin but may be due to the epoxidation of aldrin to dieldrin - a common oxidation process in many biological systems. 16 A significant correlation was observed between aldrin concentration in the adipose tissue and milk (r =
Table 2 Levels of aldrin and dieldrin in human
adipose tissue,
breast milk and blood
Downloaded from het.sagepub.com at Bobst Library, New York University on May 8, 2015
serum
in
ppb.
45
0.763; P < 0.01) and also between adipose tissue and blood serum (r 0.503; P < 0.01). However, no correlation could be found between =
the amounts of aldrin in breast milk and blood serum. A significant correlation (r = 0.503; P < 0.01) was also noticed between the dieldrin concentration in fat and blood while the accumulation of dieldrin in fat and breast milk, or in breast milk and blood serum, did not show any significant correlation. The present data clearly establishes that aldrin and dieldrin residues in India are low when compared to those reported from developed countries, and are far below the maximum recommended value of 0.0001 ppm. It also shows
that human fat is a depot for aldrin and dieldrin, as it is for many other chlorinated pesticides, and can thus be used as an index for environmental contamination. The data clearly indicates the ability of breast milk to bioconcentrate dieldrin.
Acknowledgements The authors offer their sincere thanks to Dr Arunima Sahgal, Department of Zoology, University of Delhi who helped in preparing this manuscript. The award of Senior Research Fellowship to one of us (AN) (from the University Grants Commission, New Delhi) is
gratefully acknowledged.
References 1 Howell DE.
A
case
of DDT storage in human fat.
Proceedings of Academy of Science USA 1948; 29: 31. 2 Curley A & Kimbrough R. Chlorinated hydrocarbon insecticides in plasma and milk of pregnant and lactating women. Archives of Environmental Contamination and Toxicology 1969; 18: 156-64. 3 Mes J, Doyle JA, Adams BR, Davies DJ & Turton D. Polychlorinated biphenyls and organochlorine pesticides in milk and blood of Canadian women during lactation. Archives of Environmental Contamination and Toxicology 1984; 13: 217-23. 4 Andersen JR & Orback K. Organochlorine contaminants in human milk in Denmark. AMBIO 1984; 13: 266-8. 5 Brady MN & Siyali DS. Hexachlorobenzene in human body. The Medical Journal of Australia 1972; 1: 158-61. 6 Nair A & Pillai MKK. Monitoring of hexochlorobenzene residues in Delhi and Faridabad, India. Bulletin of Environmental Contamination and Toxicology 1989; 42: 682-6. 7 Zaidi SSA, Bhatnagar VK, Banerjee BD, Balakrishnan G & Shah MP. DDT residues in human milk samples from Delhi, India. Bulletin of Environmental Contamination and Toxicology 1989; 42: 427-30. 8 Kalra RL & Chawla RP. Occurrence of DDT and BHC residues in human milk in India. Experientia 1981; 37: 4045. 9 Agarwal HC, Pillai MKK, Yadav DV, Menon KB & Gupta RK. Residues of DDT and its metabolites in human blood samples in Delhi, India. Bulletin World Health Organization 54: 349-51. 1976; 10 Dale WE, Copeland MF & Hays WJ Jr. Chlorinated insecticide in the body fat of people in India. Bulletin World Health Organization 1965; 33: 471-7. 11 Siddiqui MKJ & Saxena MC. Placenta and milk as excretory
routes of
lipophillic pesticides in women.
Human
Toxicology
1985 ; 4: 249-54. 12
Environmental Protection Agency Manual. Analytical methods for the analyses of pesticide residues in human and environmental sample. North Carolina: Health Effects Research Laboratory, 1980. 13 Snow BJ, Connor S & Koblintz R. Polychlorinated biphenyl congeners, p,p’-DDE and hexachlorobenzene in human milk in three areas of upstate New York. Archives of Environmental Contamination and Toxicology 1985; 14: 443-50. 14 Anonymous. Determination of residues of organochlorine pesticides in animal fats and eggs. Analysts 1979; 104: 42533. 15 Leoni V, d’Alessandrode Luca E & Biocca M. Aggiornamonto di dati sulla presenge dei pesticide claorogan ici nei teosuti adiposi umani. Nuovi Annali di 2. giene e Microbiologie 1977; 28: 115-24. 16 Kiigemagi U, Morrison HE, Roberts JE & Bollen WB. Biological and chemical studies on the decline of soil insecticides. Journal of Economic Entomology 1952; 51: 198-204. 17 Stacey CI & Tatum T. House treatment with organochlorine pesticides and their levels in human milk - Perth, Western Australia. Bulletin of Environmental Contamination and Toxicology 1985; 35: 202-8. 18 Griffith J & Duncan RC. Serum organochlorine residues in Florida citrus workers compared to the National Health and Nutrition Examination Survey Sample. Bulletin of Environmental Contamination and Toxicology 1985; 35: 411-17.
(Received
14 March
1991; accepted 5 August 1991)
Downloaded from het.sagepub.com at Bobst Library, New York University on May 8, 2015
1 Nair,
’Department Agricultural
P.
2 Dureja
&
M. K. K.
Fat, Milk and Blood
1 Pillai
Division of of Zoology, University of Delhi, and 2 Research Institute, New Delhi - 110012, India.
Agricultural Chemicals,
Indian
1 Aldrin and dieldrin residues were monitored in the fat, breast milk and blood serum from female residents of Delhi. 2 The average aldrin and dieldrin contents were 0.048 and 0.099 ppb in adipose tissue, 0.003 and 0.060 ppb in breast milk and 0.004 and 0.002 ppb in blood serum, respectively. 3 The older donors contained higher levels of aldrin and dieldrin in their adipose tissue. 4 Primagravidae contained more of these chemicals in their breast milk. 5 A positive correlation was observed between the aldrin concentration in adipose tissue and breast milk, and that in adipose tissue and blood serum. Similarly, a significant correlation was found between dieldrin in adipose tissue and blood serum. 6 The levels of aldrin and dieldrin were low in samples from residents of Delhi when compared to those in developed countries. Introduction The extensive use of toxic organochlorine pesticides is causing some concern. Howell’I first reported the storage of DDT in human fat. Since then, other organochlorine pesticides such as lindane, aldrin, dieldrin, heptachlor and its epoxide have been found in the breast milk, blood and adipose tissue of humans from the USA,2 Canada,; 3 Europe and Australia.~ Although there has been increased concern about the pesticide burden of the human body, only a few reports pertaining to the levels of DDT, HCH and HCB are available from India.6-1&dquo; Furthermore, little has been published about the levels of aldrin and dieldrin in human fat, blood and breast milk; only dieldrin levels in placental tissue extracts and breast milk have been cited.&dquo; This communication reports on the aldrin and dieldrin residues found in the fat, blood and breast milk of the women of Delhi.
Methods of human adipose tissue, breast milk and blood were collected from Irwin Hospital, Delhi during the 1989-90. All donors were females belonging to a lower socio-economic class and residing in those parts of Delhi exposed
Samples
to severe automobile
pollution. None of them employed and all were vegetarians. The donors of adipose tissue were between 18-42
were
years of age, while those who donated blood and breast milk were between 20-34 years. All donors had either undergone their first or second delivery. None of the donors had a history of previous abortions, or any serious recent illness. However, all of them had suffered from typhoid and jaundice in the past. Blood and breast milk samples were taken from donors with normal and healthy babies whose birthweight was over 2800 g. Adipose tissue samples were obtained from patients undergoing abdominal surgery. Breast milk samples were manually expressed from lactating mothers only after the infant was nursed (hind milk). The blood samples which were drawn from the median cubital vein of the donors, were immediately centrifuged (2500 rpm, 10 min) to obtain blood serum. In general, the Environmental Protection Agency (EPA) protocol was used for the analysis of aldrin and dieldrin.1 2-14 The adipose tissue samples (5 g) were macerated with sand and anhydrous sodium sulphate and the fat was extracted with repetitive extractions with petroleum ether and acetonitrile. 12Breast milk samples (5 ml) were shell-freezed in liquid nitrogen, lyophilized and Soxhlet extracted with n-hexane
Downloaded from het.sagepub.com at Bobst Library, New York University on May 8, 2015
44
..
,
for 6 h.’3 Blood serum samples (2 ml) were extracted with 6 ml n-hexane for 2 h in a slow speed rotatory mixer and then concentrated to 0.5 ml in a stream of nitrogen. 12 The extracted samples of adipose tissue and breast milk were concentrated in vacuo and cleaned over an alumina column with 60 ml n-hexane.’4 Residues of aldrin and dieldrin were qualitatively and quantitatively determined using a Hewlett Packard model 5890A gas chromatograph equipped with a 63 Ni electron capture detector and coupled with an integrator. A megabore capillary column 0.53 mm i.d and 10 m long, packed with 3% OV-17 was used. The operating conditions were as follows: the column temperature was initially programmed at 200°C for 3 min and was then increased at the rate of 10°C min-’ up to a final temperature of 250°C. The detector and injector were maintained at 300°C. Nitrogen with a flow rate of 50 ml min-’ was used as the carrier gas. The analytical standards of aldrin and dieldrin were obtained from the EPA. The residues of aldrin and dieldrin were further identified by using gas chromatography/ mass spectrometry (Joel model JMS DX-300).
Results The percentage recoveries of aldrin and dieldrin from spiked samples of adipose tissue, breast milk and blood samples are shown in Table 1. The average recovery was found to be more than 85%. However, the data presented in Table 2 have not been corrected for recovery. The residues of aldrin and dieldrin in human adipose tissue, breast milk and blood serum are given in Table 2. There was more aldrin and dieldrin in fat than in either breast milk or blood
Table 1 Percent recovery of aldrin and dieldrin from spiked human samples.
In addition, levels of dieldrin were twice that of aldrin. However, there was very little variation in the amounts of aldrin and dieldrin in the blood serum. No significant difference in the levels of aldrin could be observed between breast milk and blood serum. Breast milk had a 30 times higher dieldrin concentration than the blood serum.
serum.
’
Discussion The present survey indicates a considerably lower human burden of aldrin and dieldrin in women from Delhi than that reported from other countries.4.5.15,17 As with an earlier report from Australia, human fat contained more dieldrin than aldrin. Older donors contained higher amounts of chemical than younger ones. In breast milk, the mean levels of aldrin and dieldrin were found to be 0.003 ppb and 0.060 ppb, respectively, indicating bioconcentration of dieldrin in the breast. However, the levels of aldrin and dieldrin in milk were much lower than those reported for samples from Denmark or Australia. In the present study, it was also noticed that the levels of these chemicals in breast milk were dependent on the number of parturations. Women who had been delivered for the first time had more aldrin and dieldrin in their breast milk than those who had undergone their second delivery. In addition, older women with their first child had more of these pesticides in their bodies than younger ones. Unlike fat and milk, blood serum did not show any significant difference in aldrin and dieldrin levels; in addition levels in blood were much lower than those reported for samples from the USA 2,18 or
Canada.3
Although the use of dieldrin has been banned in India since 1975, its levels in adipose tissue and breast milk have been surprisingly higher than those of aldrin. Hence, whatever dieldrin has been detected may not actually be environmental in its origin but may be due to the epoxidation of aldrin to dieldrin - a common oxidation process in many biological systems. 16 A significant correlation was observed between aldrin concentration in the adipose tissue and milk (r =
Table 2 Levels of aldrin and dieldrin in human
adipose tissue,
breast milk and blood
Downloaded from het.sagepub.com at Bobst Library, New York University on May 8, 2015
serum
in
ppb.
45
0.763; P < 0.01) and also between adipose tissue and blood serum (r 0.503; P < 0.01). However, no correlation could be found between =
the amounts of aldrin in breast milk and blood serum. A significant correlation (r = 0.503; P < 0.01) was also noticed between the dieldrin concentration in fat and blood while the accumulation of dieldrin in fat and breast milk, or in breast milk and blood serum, did not show any significant correlation. The present data clearly establishes that aldrin and dieldrin residues in India are low when compared to those reported from developed countries, and are far below the maximum recommended value of 0.0001 ppm. It also shows
that human fat is a depot for aldrin and dieldrin, as it is for many other chlorinated pesticides, and can thus be used as an index for environmental contamination. The data clearly indicates the ability of breast milk to bioconcentrate dieldrin.
Acknowledgements The authors offer their sincere thanks to Dr Arunima Sahgal, Department of Zoology, University of Delhi who helped in preparing this manuscript. The award of Senior Research Fellowship to one of us (AN) (from the University Grants Commission, New Delhi) is
gratefully acknowledged.
References 1 Howell DE.
A
case
of DDT storage in human fat.
Proceedings of Academy of Science USA 1948; 29: 31. 2 Curley A & Kimbrough R. Chlorinated hydrocarbon insecticides in plasma and milk of pregnant and lactating women. Archives of Environmental Contamination and Toxicology 1969; 18: 156-64. 3 Mes J, Doyle JA, Adams BR, Davies DJ & Turton D. Polychlorinated biphenyls and organochlorine pesticides in milk and blood of Canadian women during lactation. Archives of Environmental Contamination and Toxicology 1984; 13: 217-23. 4 Andersen JR & Orback K. Organochlorine contaminants in human milk in Denmark. AMBIO 1984; 13: 266-8. 5 Brady MN & Siyali DS. Hexachlorobenzene in human body. The Medical Journal of Australia 1972; 1: 158-61. 6 Nair A & Pillai MKK. Monitoring of hexochlorobenzene residues in Delhi and Faridabad, India. Bulletin of Environmental Contamination and Toxicology 1989; 42: 682-6. 7 Zaidi SSA, Bhatnagar VK, Banerjee BD, Balakrishnan G & Shah MP. DDT residues in human milk samples from Delhi, India. Bulletin of Environmental Contamination and Toxicology 1989; 42: 427-30. 8 Kalra RL & Chawla RP. Occurrence of DDT and BHC residues in human milk in India. Experientia 1981; 37: 4045. 9 Agarwal HC, Pillai MKK, Yadav DV, Menon KB & Gupta RK. Residues of DDT and its metabolites in human blood samples in Delhi, India. Bulletin World Health Organization 54: 349-51. 1976; 10 Dale WE, Copeland MF & Hays WJ Jr. Chlorinated insecticide in the body fat of people in India. Bulletin World Health Organization 1965; 33: 471-7. 11 Siddiqui MKJ & Saxena MC. Placenta and milk as excretory
routes of
lipophillic pesticides in women.
Human
Toxicology
1985 ; 4: 249-54. 12
Environmental Protection Agency Manual. Analytical methods for the analyses of pesticide residues in human and environmental sample. North Carolina: Health Effects Research Laboratory, 1980. 13 Snow BJ, Connor S & Koblintz R. Polychlorinated biphenyl congeners, p,p’-DDE and hexachlorobenzene in human milk in three areas of upstate New York. Archives of Environmental Contamination and Toxicology 1985; 14: 443-50. 14 Anonymous. Determination of residues of organochlorine pesticides in animal fats and eggs. Analysts 1979; 104: 42533. 15 Leoni V, d’Alessandrode Luca E & Biocca M. Aggiornamonto di dati sulla presenge dei pesticide claorogan ici nei teosuti adiposi umani. Nuovi Annali di 2. giene e Microbiologie 1977; 28: 115-24. 16 Kiigemagi U, Morrison HE, Roberts JE & Bollen WB. Biological and chemical studies on the decline of soil insecticides. Journal of Economic Entomology 1952; 51: 198-204. 17 Stacey CI & Tatum T. House treatment with organochlorine pesticides and their levels in human milk - Perth, Western Australia. Bulletin of Environmental Contamination and Toxicology 1985; 35: 202-8. 18 Griffith J & Duncan RC. Serum organochlorine residues in Florida citrus workers compared to the National Health and Nutrition Examination Survey Sample. Bulletin of Environmental Contamination and Toxicology 1985; 35: 411-17.
(Received
14 March
1991; accepted 5 August 1991)
Downloaded from het.sagepub.com at Bobst Library, New York University on May 8, 2015
