Document 109YrB9n5YoL7yYQ8epGyQDL5

Pesticide Residues and Polychlorinated Biphenyl Levels in Diets, Urine, and Fecal Matter o( Preadolescent Girls (36347) Nelson O. Price, R. W. Young, and Jean K. Dickinson (Introduced by G. E. Bunce) Department of Biochemistry end Nutrition, Virginia Polytechnic Institute and Stale University, Blacksburg, Virginia Z40O1 Description of six earlier Southern Regionaf Studies with preadolescenl girls 7 to 9 years of age have been published (1-3). The 1970 study, including overall objectives, sub ject management, exact dietary compositions, and methodology of sample collection, was reported (4), Also included in some of the past work in these metabolic studies are the determinations of certain trace minerals in lhe\liets, urine, and feces (S--7). This work summaries the screening of weekly composite samples of diets, urine, and fecal matter for pesticide residues. This is the first work reported on polychlorinated biphenyl (PCB) residues from a human met abolic study where controlled conditions of the diets and sample collection were estab lished. The residue levels, including PCB, may hopefully be of great value as a basis for further invesligafion in this field with hu man subjects. The chlorinated aromatic compounds have been used extensively in industry since 1929 (8). In 1952, a bovine hyperkeratosis disease appeared in a number of cattle herds in Virginia. Highly chlorinated naphthalene was established as the causative agent (9) for this disease. This compound was found in certain petroleum ' products (lubricants) es sential for various types of farm machinery (10). The petroleum Industry eliminated the highly chlorinated naphthalene from their products and the hyperkeratosis disease of cattle disappeared in VA. Recently, Vos and Beems (11) have shown that the application of PCB to the skin of rabbits produced hy perplasia and hyperkeratosis which resembled the changes observed in animals exposed to the chlorinated naphthalenes. Polychlorinated biphenyls are structural analogs of the chlo rinated naphthalenes and are used as lubri cants (11). It would be reasonable to suggest a more vigorous effort to identify the occur rence of PCB in petroleum products. Procedure for sample collection and diet composition. Two replicate nutrition studies, each with sixteen 7- to 9-year-old girls, were conducted at Blacksburg, VA, during June, July and Aug., 1970. In each study, the first 8 days (two 4-day periods) were used for adjustment to the basal diet. The diet em ployed was considered typical of the foods consumed by lower socioeconomic groups in the southeastern United States. Each diet provided about 2000 kcal of energy, 24 g of protein, and was marginal in vitamin A, thia mine, riboflavin, and niacin equivalents. Af ter the adjustment period, the subjects were randomly assigned within weight groups to four modifications of the basal diet and were maintained on them for two periods, each consisting of 6 consecutive days. The modifi cations consisted of variations in calcium (300-1300 g/day) or nitrogen (L-lysine-HCl, 1.139 g; L-threonine, 0.63 g; L-methionine, 0.189 g; or ammonium citrate to provide an equivalent amount of nitrogen), but fhe data from each treatment have been pooled for this report since there were no patterns evi dent as a function of dietary variation. Composites of food, feces, and urine were made as follows: The complete diet for each day, per subject, was weighed and adjusted to 2000 g with ion-free water. From this daily composite, portions were transferred into a liter plastic container, capped, and frozen. At the end of each period, the daily aliquots were thawed, blended thoroughly, and urn- 1280 TABIj PCU typ 1242 1254 1200 1242, 125 (00% r * Aroclt pied. The 24 sured for eac made to 200 which a coni feces for eai weighed; ion the net weigh 1000 g for th< for the two samples for t! in Nalgene p! diets, 10 g of the urine wer lions and detei Methods i Microtek 220 and PCB dr inated biphen> DDT and its Armour and B ery of TCB urine, and feca Sensitivity for 0.001 ppm. The procedt lowed for the d< the feces and t in the initial e (a) Transfer a into a 250-ml o imately 25 ml of acetonitrile and fuge for 20 min glass wool into peat the extrac trile, shake, cen (c) Flash evapi removed, leavin Measure the wa P.<5 .~./V\.&. ido/. I osw 031201 STLCOPCB4015163 <`y, e chlo. lubrisuggest occur- id diet studies, s, were June, he first for et em foods >ups in h diet M g of thiats. Afis were ups to d were , each modifialcium >e*HCl, tionine, dde an )e data ed for ns evi- e were >r each ijusted s daily into a 'jtn. At liquols 1 sam- PESTICIDE AND PCB RESIDUES 1281 TABLE I. Rocovory (%) of Polychlorinated Blphonyla (PC)* Jn 2>iot-n, Focoe, and Urine, Diets (%) Focee (%) Urine (%) PCB types 1 2 8 Av 1 2 8 At 1 2 8 At 1242 62 68 72 67 1254 mo 66 68 72 68 62 68 65 65 1242,1254 68 67 60 68 (60% mixture) 70 68 72 70 65 60 70 68 68 61 64 64 70 66 67 67 71 68 60 68 60 68 ' 62 66 66 60 70 68 65 62 68 66 Arochlor (PCB) t/poe, 1242, 1254, 1260 supplied by Monsanto Chemical Co., 8t. Louis, MO. pled. The 24-hr urine collections were mea sured for each subject and the total volume made to 2000 ml with ion-free water, from which a composite sample was taken. The feces for each subject were collected and weighed; ion-free water was added to bring the net weight of the composite to a total of 1000 g for the two 4-day periods and 15000 g for the two 6-day periods. All composite samples for the residue analyses were stored in Nalgene plastic bottles. Ten grams of the diets, 10 g of the fecal matter, and 50 ml of the urine were used for the residue extrac tions and determinations. Methods and sample preparation. A Microtek 220 GLC was used for the residues and PCB determinations. The polychlor inated biphenyls (PCB) were separated from DDT and its analogs using the method of Armour and Burke (12). The average recov ery of PCB from spiked samples of diet, "'urine, and fecal matter was 67.0% (Table I). Sensitivity for all residue determinations was 0.001 ppm. The procedure of Samuel (13) was fol lowed for the determination of the residues in the feces and diet, with some modifications in the initial extractions and preparations: (a) Transfer a 10-g sample of fecal matter into a 250-ml centrifuge bottle. Add approx imately 25 ml of ion-free water and 150 ml of acetonitrile and shake for 1 min. (b) Centri fuge for 20 min at 1500 rpm. Decant through glass wool into 500-ml Erlenmeyer flask. Re peat the extraction with 100 ml of acetoni trile, shake, centrifuge, and decant as before, (c) flash evaporate until, the acetonitrile Is removed, leaving only the water layer, (d) Measure the water layer, transfer to a 500-ml separatory funnel, and add an equal amount of methylene chloride. Shake well and allow to settle for 15 min. Transfer the methylene' chloride layer through an anhydrous Na2SO< column into a 500-ml Erlenmeyer flask. Ex tract with three more portions of methylene chloride, add to the original extract, evap orate to dryness on a flash evaporator, (e) Add 50 ml of ethyl acetate to the sample and pour onto a column which is prepared by making 15 g of 1 part acid-washed attapulgus clay and 1 part Celite 545 (John Manville) topped with anhydrous NajSOi. Attapulgus clay is substituted in place of charcoal (Norit A) (14) in cleanup!section. The procedure of Teasley and Cox (14) was followed for the determination of res idues in urine. The PCB's were separated from DDT and its analogs as previously staled. Results of the residue analysis are shown in Table II and are expressed: (^tg/day/subject) and (maximum percent age of intake excreted). Summary and Conclusions. The pesticide residue levels in the diets, urine, and fecal matter were not affected by supplementation of the diets with amino acids, ammonium citrate, or calcium lactate. Pesticide residues were found in all samples of the diets, urine, and feces. With the exception of the chlor inated cyclodienes, excretion of all residues was greater in the feces, suggesting the feces as the primary route of excretion. Excretion of the PCB's was predominately through the feces with 6.2% of the total excretion appear ing in the urine. With the exception of DDT and its metabolites, excretion of residues was 9 to 12%. Maximum excretion of DDT and its metabolites was approximately 32%. DSW 031202 STLCOPCB4015164 1 >282 PESTICIDE AND PCB RESIDUES TAlll/B II. Ktimk*' nml Muxiniiiiii IVrc.mitngc of Intake of Potlel<lc Residue* mill Pnlyrldurinnted Diphenyl (PCB) I/ovcIh in Dicta, Urine, mid Kocnl Matter of Proadolesrent Girin, 7 to 0 Years of Age." Lindane Keltlinne Cliloriimted e.ye.lodiene pesticides* Arochlor (polye.hlor. blphonyl)* DDT and metabolites* Sample - (s'B/dnj'/ Max typo subject) (%) (s-gAIny/ Max subject) (%) Oig/day/ Max subject) (%) (fig/dny/ Max subject) (%) (A/d*y/ Max subject) (%) Dint Uriito Feces Tr*-1.8 Tr-0.00 Tr-0.12 3.12 6.21 Ti-1.8 Tr-0.00 Tr-0.12 3.12 6.21 Tr-1.8 TM1.10 Tr-0.12 6.24 6.21 Tr-840.0 Tr-6.8 0.69 Ti-87.0 10.86 Tr-18.0 Tr-2.1 11.80 Tr-3.7 20.74 16. Dr' Conlani. ' 17. Gu (1970). 18. Ha I., Arch I 19. Hu Technolo: rds.). 2n< New Yor 32 subjects, 40 days (2 duplicate experiments); Tr = trace. 1 Aldriu, dicldrin, endriu, lieptnrlilor, hcptnehlor epoxide. Bum of PCB type* I (1242), II (1854), III (1200). 4 DDT mid metabolites, p.p'-PDB, p,p'-PDP, 0,p'-DDD, 0,p'-DDT, Traco (<.001 ppm). I'CB's were found in all eight diets analyzed. and Foods at Virginia Polytechnic Institute and Four contained traces and four contained 200, 420, S80 and 840 pg/day/subject. The FDA has not yet established tolerance levels for PCB. Albert Kolbye, deputy director for the FDA's Bureau of Foods has, however, recently slated that PCB is regarded as mod erately toxic but certainly less toxic than DDT. He suggested that 150-300 pg/day would be within the safe range as far as temporary dietary intake is concerned (15). Eighty-eight percent of the ingested PCB's were not excreted. Presumably these have been retained in the body. The comparable Stale University, Blacksburg, VA for providing the samples and other technical data; technicians Patri cia L. Marshall and Sylvie R. Bcalty, who prepared and extracted the samples (or analysis; and Dr. R. E. Webb for his suggestions in the preparation of the data for this article. 1. S. Regional Nutr. Res. Proj. (S-28) Bull. No. 64, Blacksburg, VA (1959). 2. S. Regional Nutr. Proj. (S-28) Bull. No. 94, Blacksburg, VA (1962). 3. S. Regional Nutr. Proj. (S-64) Bull. No. 129, Blacksburg, VA (1967). 4. S. Regional Nutr. Proj. (S-64) Bull. No. 170, Blacksburg, VA (1970). percentage for DDT and its metabolites was 5. Engel, R. W., Miller, R. F., and Price, N. O., in 67.5%. It is difficult to explain the presence of high levels of PCB found in these diets though PCB has been identified in such sources as sediments, river water, sewage out falls, ink, newsprint, plastic bags, plastic pro ducts, poultry and animal tissues (16-23). The plastic containers used for storage may be a possible source of contamination, though "Zinc Metabolism" (A. S. Prasad, cd.), Chap. 18, p. 326. Thomas, Springfield, IL (1966). 6. Engel, R. W., Price, N. O., and Miller, R. F., J. Nutr. 92, 197 (1967). 7. Price, N. O., Bunco, G. E., and Engel, R, W., Amer. J. Clin. Nutr. 23, 258 (1970). 8. Henninger, D., The National Observer, Oet. 9, 9 (1971). 9. Bell, W. B., Va. J. Sci. 3, 169 (1952). 10. Engel, R, W., Price, N. O., Linkous, W. N., this possibility has not been tested. The and Bell, W. B., Va. Agr. Exp. Sta. Tech. Bull. 121 diets did consist of 20% fat which is consid (1953). ered normal for low income diets of the southeastern United States. Fats in the diets i of the subjects before the study were primari\ ly animal, whereas fats in the prepared exIperimental diets were chiefly vegetable. 11. Vos, J. G., and Bcems, R. B., Toxicol. Appl. Pharmacol. 19, 617 (1971). 12. Armour, J., and Burke, J., J.A.O.A.C. 53, 761 (1971). 13. Samuel, B. L., J.A.O.A.C. 49, 346 (1966). 14. Teasley, J. I., and Cox, W. S., US. Dept. The author* gratefully acknowledge the Southern H.E.W., 1,11C, 4b (1965). J Regional Nutritional Project (S-64) and It* Techni 15. Kolbye, A. C., Chem. Eng. News 49, 12 cal Committee; the Department of Human Nutrition (1971). DSW 031203 s'gwstll b STLCOPCB4015165 iilvrlilurinntcil Ours of Ago.* DDT and metabolites* ig/'lny/ Max .ilijrat) (%) 0-18,0 Tr--2.1 11.80 Pr-3.7 20.74 1 / PESTICIDE AND PCB RESIDUES J 283 16. Devos, R. H., and Peel, E. W., Bull. Environ. Contam. Toxicol. 6, 1648 (1971). 17. Gutafson, C. G., Environ. Sci. Technol. 4, 814 (lMO). 18. Hayet, J. W., Jr., Dale, W. E., and Pirkle, C. I., Arch. Environ. Health 22, 119 (1971). 19. Hubbard, H. L., in "Encyclopedia of Chemical Technology" (R. E. Kirk and D. F. Othmar, eds.), 7nd ed., Vol. J, p. 289. Wiley (Interadence), New York (1964). 20. McCunc, E. L., Savage, J. E., and O'Dell, B. L., Poultry Sci. 41, 295 (1962). 21. Peakall, D. B., and Lincer, J. L., Bioscience 20, 9S8 (1970). 22. Reynolds, L. M., Bull. Environ. Contam. Tox icol. 4, 128 (1969). 23. Risebrough, R. W., Rieche, P,, Herman, S. G., Peakall, D. B., and Kirren, M. N., Nature (London) .220, 1098 (1968). Received Nov. S, 1971. P.S.E.B.M., 1972, Vol. 139 institute and providing the hnicians PatriBrall}', who r analysis; and st ions in the 28) Bull. No. Bull. No. 94, Bull. No. 129, Bull. No. 170, Price, N. 0,, in . ed.), Chap. .6), Idler, R. F., J. Engel, R. W., rver, Oct. 9, 9 nkous, W. N., ech. Bull. 121 Toxicol. Appl. 3.A.C. S3, 761 (1966). US. Dept. News 49, 12 v \S / \.' DSW 031204 STLCOPCB4015166