Document 3e7OKyv8q6k4vBODpQQmRzObE

PROPOSAL FOR A HISTORICAL PROSPECTIVE MORTALITY STUDY Of POLYCHLORINATED BIPHENYL EXPOSED WORKERS BACKGROUND The polychlorinated biphenyl* (PCB'a) have attracted much attention since the late 1960's when technological developments in gas chromatography and mass spectrometry made the detection of PCB's at low levels in the environment possible. PCB's have been in use sine# 1929 in transformers, lubricants, "carbonless" duplicating paper, fireproof sealante, plastics, adhesives, and many other products. Their resistance to biodegradation, photodegradation, and oxidation made them invaluable in scores of uses but also made them more stable as residual pollutants when released into the environment. PCB's are highly lipid soluble compounds. Lipid solubility enables a proportion of food chain PC8 residues to be stored in the fat depots of the body. The first reports of PCB's in the environment came from Sweden, where Jensen (1966)1 identified residues of PCB's . in fish from various Swedish waters. Risebrough et al (1968) found PCB and DDT in marine ecosystems of the Pacific Ocean. Early reports of wildlife effects included egg shell thinning and baby seal effects, but subsequent studies have failed to conclusively show PCB's as the causative- egent. The first report of possible PCB effects on human* ocourred in 1966, when over 1,000 Japanese were involved in an epidemic of Yusho, an acnelike skin disease which was traced to rice oil contaminated with PCB's. However, it was later shown that the rice oil also contained a high level of polychlorinated dibenzofurans (PCDF'e), possibly due to us* of the contaminating PCB as a high temperature heat transfer medium It was shown that PCDP's are relatively more concentrated in the liver, MOMS 056199 2- and the investigators' findings olaarly indicated the necessity to pay greater attention to PCDF'e for clarification of the nature of Yusho.3 While acute effects from relatively high PCB exposure levels are known and can be avoided by appropriate control measures, attention has more recently been focused on the chronic effects . resulting from long-term low levels of exposure to PCB's. Animal tests have proved to be equlvocable. The Environmental Protection Agency considers PCB's to be suspect'oareinogens based on its own animal tests. Monsanto's extensive two-year chronic oral toxicity testa of Arochlor 1242, 1254, and 1260 in both rata and beagles at dose levels of 1, 10, and 100 ppm concluded that PCB's were non- oarclnogenic. Hyperplasia, hypertrophy, and several benign tumors were observed in the livers of the rats, but there was no evidence of malignancy in any of the test animals.* Other investigators have shown liver tissue effeots ranging from hypepplasia to well-differentiated hepatocellular carcinomas in rats or mice fed with various PCB-containing diets. For example, Kimbrough et al (1972)5 fed rata with Arochlor 1254 and 1260, and found hyper trophy, lipid accumulation, and adenofibrosia in liver tissue, but found no malignancy. The same investigator fed ArochIorl254 to mice for 11 months and reported the induction of malignant hepatomas in the test animals.* The recent National Cancer Institute report 7 of its carcinogenicity tests on Arochlor 1254 concluded that the substance was not carcinogenic in Fischer 344 rats under the conditions of the bioassay. However, they allude to the non-significant occurrence of gastrointestinal tract carcinomas and hepatocellular proliferative lesions in their test animals as possibly being related to the Arochlor diet. There have been very few epidemiological studies of human populations HONS 056200 -3 exposed to low levels of PCB's over a long period, The Mobil oil Company released a study In 1976 conducted by Dr. Anita Bahn8 in which a significant number of melanomas and pancreatio cancers were found In exposed chemical workers. While these results received publication, A the study was severely criticized due to deficiencies in the identifi cation of the study cohort, failure to identify other potentially carcinogenic chemicals to which the workers and research personnel were exposed, and other problems which left the results of the Mobil study highly in doubt. A preliminary study of PCB-exposed workers was begun at Monsanto shortly after the Mobil study in response to the need for a valid epidemiological study of the effect of PCB's on humans. Hie study focused on workers at the Krurmrlch Plant, where PCB's were manufactured from 1936 until production ceased in 1977. No melanomas or pancreatic cancers were found, but an excess incidence of lung cancer deaths was seen. However, due to the preliminary nature of the study, several necessary ' steps to complete the epidemiology study remained unfinished, such as following up on the mortality experience of over 30% of the study cohort who ware lost-to-follow-up. In addition, no minimum limits on exposure time were used in the preliminary study, thus, no attempt was mads to distinguish between the workers exposed for years the worker exposed for a few days. The investigators involved in the preliminary Monsanto PCB study realized these deficiencies and advised that the study population should be further defined and evaluated so that a valid, comprehensive report could result. Therefore, due to the preliminary nature of the epidemiological data available and the inconclusive toxicological data, it is necessary to pursue further epidemiologic study in order to evaluate the chronic effects of PCB-exposure to man with more complete vital information on a verified, exposed worker cohort. HONS 056201 -4- PURPOSE OF THE STUDY This epidemiological study will investigate the mortality xperience of workers exposed to polychlorinated biphenyls in their manufacture. Through studying this cohort wt intend to (1) identify groups of workers that have PCB exposure of a sufficient latency to demonstrate any chronic health effects, and (2) analyze the mortality experience of these workers with attention given to malignant neoplasms especially of the lung, liver, skin, and pancreas. - . STUDY POPULATION The Monsanto plants involved in the U.S. manufacture of PCB's are the W.6. Krummrich plant in Sauget, Illinois, and the Anniston, Alabama plant. Since the preliminary study, upon . which the proposed study is based, involved the Krummrich plant alone, the proposed study will deal solely with the Krummrich workman population^ . * i The study population from the Krummrich plant consists of all male waged employees who have worked at least six months in a job involving exposure to PCB after December 31, 1944,and prior to December 31, 196S. This population includes active, retired, and terminated employees. Through interviewing plant personnel, it was determined that exposure levels of workers involved in the manufacture of PCB's did not vary considerably) therefore, all waged workers involved in the manufacture of PCB's will be considered to have a common exposure level. HONS 056202 Work hletorlea for all part and praaant Krumrieh employee! have baan obtalnad and tcreened for PCB~departnent work by tha haa aiau oaan euppnea to uie men, ana tht computer Hating baa baan croas-chackad with tha hand work hlatory aaarch In ordar to inaure eompleteneaa. It ahould b* notad that aome workere, e.g., maintenance, may hava had algnlflcant axpoauraa to PCS'a, but ara not lncludad In tha atudy dua to expoaure verification difficultiaa. por aach worker tha following Information haa been obtained* name, aoolal aacurity number, amployaa number, race, aax, data of birth, data of hire, data of termination (If applicable), and aa detailed a work hlatory aa la available. For thoaa known to hava died, tha data and place of death, and a copy of tha death certificate ara being obtalnad. MOWALITV FOLLOW-W 1. All active employea,and ratlreaa who ara currently receiving panalon checka will be returned to be alive aa of December 11, 1977. 2. All pereone known to have died on or after January 1, 1979, will be returned to be alive for the purpoeer of thle atudy. 1. Pereonnel foldere and panalon file! will ba eearched for death oartiflcatea or for pertinent information about the decedent if no death certificate la preaent. 4. The name of pereone for which there la no information on vital atatue will be reviewed with plant peraonnal for any information which they can provide, HONS 056203 5. Other follow-up procedure*, Such at Veteran's Administration files, folk Directories searches, telephone directories searches and state driver's license files will be used, if necessary, to determine the vital status of lost-to-follow-ups. (. A State death certificate will be obtained for every person known to have died. 7. An experienced, trained nosologist will review and code ' all death certificates for the underlying cause of death to the International Classification of Diseases. Seventh Revision. ANALYSIS .. Itie data will be analysed using the modified life-table method. In this method of analysis,the age, race, cause, time-specific mortality rates in a standard population are applied to person- years classified by age, race, and time-specific subgroups of the worker population at risk (i.e. exposed) over the years Of observation intervals. A standard mortality ratio will be calculated as the ratio of the sum of the observed deaths to the sum >of the expected deaths xlOO. The person-years of observation will be calculated by year of Observation and age at observation as shown in Appendix A. Person-years of observation will be accumulated from date of first exposure to PCB's to the earliest of the following eventsi 1) date of death, 2) December 31, 1977, the cut-off ascertainment date, or 3) date at which the study member was lost-to-follow-up. HONS 056204 -7The expected number of death* will be calculated using United States annual mortality rates, or county rates, if available, for every year from 1945-1977. if annual county mortality data is unavailable, the 1950-69 expected numbers of cancer deaths based on U.S. rates will be adjusted by cause for the local county death rates according to the ratio of the local county rate to the U.S. rate using U.S. Cancer Mortality by Countyi 1950-69. A standard mortality ratio will be calculated for certain selected causes of death (Appendix B). David C. Musch KONS 056205 APPENDIX A PERSON--TEARS OT OBSERVATION BT TEAR OF OBSERVATION ADD AGE 15-24 1945-49' ? '1950-54 1955-59 . 1960-64 . 1965-69 . 1970-74 25-34 35-44 AGE 45-54 55-64 65-74 75-84 85 +....... HONS 056206 /Vtzaoix. B Knnrlrh KB Study Cause of Death with I.C.D. Mosher (Seventh Revision) Malignant neoplaaas (140-205) Succal Cavity and Pharynx (140-148) Digestive Organs and Peritonea* (130-156A, 157-159) Biliary Passages and liver (155-156A) Other Parts of Digestive Systen (Residual) Respiratory Systen (160-164) Bronchus, Lung, and Trachea (162-163) Other Parts of Respiratory Systen (Residual) Cenito-Urlnary Systen (177-181) Leukemia and Aleukenls (204) Lynphosarcona and Other Heoplaans of lynphetlc and Hematopoietic Tissues (200-203, 205) All Other Heoplasns (Residual) Diseases of the Cardiovascular Systen (330-334, 400-468) Diseases of the Respiratory Systen (470-527) Diseases of the Digestive Systen (530-587) All Other Diseases (Residual) All External Causes (E800-E998) Expected P.H.R. REFERENCES 1. Jensen, S., New Scientists, Vol. 32, p. 612.. 2. Risebrough, R. K. et al, "Chlorinated Hydrocarbons in Marine Ecosystems," Nature, Vol. 220, No. 1098, 1968. 3. Nagayama, Junya et al, "Determination of Chlorinated Dibenzo- furans in Kanechlors and Vusho Oil," Bulletin of Environmental Contamination and Toxicology, Vol. 15, No. 1, 1976. 4. Report to Monsanto Company - "Two Year Chronic Oral TOxiclty With Arochlor 1242 in Albino Rats and Beagles" (Unpublished), November 12, 1971 and March 24, 1975. 5. Kimbrough, R. D., and Linder, R. E., and Gaines, T. B., "Morphological Changes in Liver of Rate Fed With Polychlorinated Biphenyls," Archives of Environmental Health, Vol. 25, pp. 354-64, 1972. 6.. itimbrough, R. D., and Linder, R. E., "Induction of Adenofibrosis and Hepatomas of the Liver in BALB/oo Mice by Polychlorinated Biphenyls (Arochlor 1254)", Journal of the National Cancer Institute, Vol. 53, No. 2, pp. 547r552, 1974. 7. National Cancer Institute, BIOASSAY OF AROCHLOR 1254 FOR POSSIBLE CARCINOGENICITY, Technical Report Series No. 38, 1978, DHEW Publication No. (NIH) 78-838. 8. Bahn, Anita K. , REPORT ON PAULSBORO, N.J. MOBIL OIL PLANT STUDY. April 27, 1976. (Unpublished) 9. Lawrence, Charles, "PCB7 and Melanoma," New England Journal of Medicine, Vol. 296, No. 2, January 13, 1977. HONS 056208 PINAL REPORT OF THE SUBCOMMITTEE ON THE HEALTH EFFECTS OF POLYCHLORINATED BIPHENYLS AND POLYBROMINATED BIPHENYLS JULY 19 7E. DEPARTMENT OF HEALTH, EDUCATION AND WELFARE WASHINGTON, I'.C. Attachment 1 7 1 RONS 056209 hRKATUM Table numbers proceed from 9 to 11. There is no Table 10. HONS 056210 CONTENTS Members and Consultants, Subcommittee on ths Haalth Effects of PCBs and PBBs......................................................... v Subgroups and Their Members.......................................................vll Purpose and Specific Objectives.................................................... INTRODUCTION........................................................................................ 1 GENERAL SUMMARY AND CONCLUSIONS...................................................A CHEMISTRY........................................................................................ 4 METABOLISM AND BIOCHEMICAL TOXICITY ..................................... 9 ANIMAL TOXICOLOGY.......................................................................12 HUMAN EXPOSURE............................................................................ 17 GENERAL RECOMMENDATIONS.................................................................25 DETAILED RECOMMENDATIONS...............................................................27 CHEMISTRY .......................................................................................27 METABOLISM AND BIOCHEMICAL TOXICITY................................... 28 ANIMAL TOXICOLOGY....................................................................... 30 HUMAN EXPOSURE................................. 31 CHEMISTRY OF PCBs AND PBBs........................................................... 34 Introduction.................................................................. 34 Chemistry of Chlorinated Biphenyls......................................33 A. Synthesis of Chlorinated Biphenyls..........................35 B. Non-metabollc Alteration of Chlorinated Blnhenyls...........................................................................37 Oxidation........................................................................... 37 Hydrolysis and Alcoholysis..........................................38 Photochemistry............................................................... 38 1 HONS 056211 C. Chemical Structure Related to Occurrence, Fate and Effect* of Chlorinated Biphenyls....................42 1). Determination of PCB Residues,..............................18 E. Chlorinated Dlbentofurans...........................................61 Chemistry of Brominated Biphenyls.............................................. 68 A. Comparison of Brominated Biphenyl with Chlorinated Biphenyls............................................. 68 B. Non-metabollc Alteration of Brominated Biphenyls ................................... 70 Oxidation and Hydrolysis......................................... 70 Photochemistry................................................................70 C. Brominated Biphenyls and Determination of Residues................................................................... 72 D. Possibility of Brominated Dlbenaofurane In Commercial PBB Mixtures..................................... 76 METABOLISM OF BIOCHEMICAL TOXICITY OF PCBs AND PBBs........ 75 Effects of PCBs on Biochemical Function ...................... 75 The Comparative Biochemical Toxicity of Aroclorl254 and Flreaaatar BP-6............................................................... 83 Metabolism of PCB and PBB Mixture ................................... 87 The Metabolism of Individual PCBs .................................... 96 ANIMAL TOXICOLOGY ......................................................................... 100 Introduction.................... '..................................................... 100 Acuta Toxicity ....................................................................... 100 Polychlorinated Biphenyls ........................................ 100 Polybrominatad Biphenyls..................................... 102 Sub-Acute and Reproductive Effects of PCBs, PBBs and Chlorinated Dibsnsofurana .................................... 107 M0NS 056212 . A. Mammalian Sub-Acute Effects of PCBs..................107 B. Avian Sub-Acute Effects of PCBs................ ..,.*113 C. Mammalian Reproductive Effects of PCBs....... 114 D. Mammalian Sub-Acute and Reproductive Effects of PBBa.................................................... H8 E. Chlorinated Dlbenzofurans - Mammalian Toxicity........................................................... -......... 122 F. Chlorinated Dlbenzofurans * Avian Toxicity ....................................................................... 123 G. Broroinated Dlbenzofurans - Mammalian Toxicity................................................... 124 H. Longterm Toxicity Including Tumorogenesl* .... 124 Chlorinated Biphenyl............... Decachloroblphenyl ................................. 124 129 Chlorinated Dlbenzofurans and Chlorinated Naphthalenea................................................. 129 Immunosuppressive Effects of PCBs snd PBBa ... 129 Mutagenicity and Teratology .................................. 130 Polychlorinated Biphenyls....................... 130 Teratology..................................... ..131 Chlorinated Dlbenzofurans ................................. 132 Toxicity of Chlorinated Naphthalenes ................ 133 HUMAN EXPOSURE TO POLYCHLORINATED BIPHENYLS.................140 Introduction ............................................................................. 140 Dietary r<posure to PCBs * *....................... 140 PCS Residue In Fish................................... 144 PCB Exposure in Rice Oil......................... .....160 Occupational Exposurs to PCBs........................ *................. 160 MONS 056213 Air and Water Exposure to PCBs 182 Realduea of PCB in Human Tissue and Milk......................184 HUMAN EXPOSURE TO POLYBROKINATED BIPHENYLS........................ 187 Introduction ....................... 187 Human Exposure ................ 188 iv HONS 05621*1 subcommittee on the health effects OF POLYCHLORINATED BIPHENYLS AND POLYBROMINATED BIPHENYLS Chairperson Albert C. Kolbya, Jr., M.D. Food and Drug Administration Members (M) and Consultants (C) Dr. John Buckley (C) Environmental Protection Agency Mr. Jerry Burke CM) Food and Drug Administration Dr. Frank Cordle (M) Food and Drug Administration Mr. Paul Cornelluasan (M) Food and Drug Administration Dr. David Firestone (M) Food and Drug Administration Dr. Laurence Flshbeln (M) Food and Drug Administration Dr. Gary Flamm (M) National Cancer Institute - Dr. Georgs Fries (C) U.S. Department of Agriculture Mr. Albart Gardner (M) Food and Drug Administration Dr. Larry Garthoff (M) Food and Dr- % Administration Ms. Helga Cerstnsr (C) Oak Ridge National Laborato.y Dr. Joyce A. Goldstein (C) Environmental Protection Agency Mi. Bitty Hscklay (C) National Marina Fisheries Servlca Dr. Charlas F. Jellnek (M) Food and Drug Administration Dr. Louis Kaaza (M) Food and Drug Adninlatration Dr. Ranata Kimbrough (M) Cantor for Dlaaasa Control Mr. Thomas E. Kopp (C) Envlronmantal Protactlon Agancy Dr. Yuoh Ku (M) Food and Drug Administration Dr. Richard L. Lahman (C) Dapartmant of Commarca Dr. William L. Marcus (C) Envlronmantal Protactlon Agancy Dr. H. 8. Matthavs (M) National Inatltuta for Envlronmantal Haalth Sdsncas Dr. J. D. McKlnnay (M) National Inatltuta for Envlronmantal Bsalth Sclancas Dr. Joaaph McLaughlin (C) Consumer Product Safety emailsalon Dr. John A. Moors (M) National Inatltuta for Envlronmantal Haalth Sclancas Dr. Irwin Pomaranta (M> Food and Drug Administration Dr. Richard A. Rhoden (M) National Inatltuta for Occupational Safaty and Haalth Mr. John A. Roach (M) Food and Drug Adninlatration Dr. Samual I. Shlbko (M) Food and Drug Adninlatration Dr. Raymond E. Shapiro (M) Food and Drug Administration vl MOMS 056216 Or. Richard H. Teake (M) Food and Drug Adalnlatratlon Dr. ViUlai Trotter (H) Food and Drug Adalnlatratlon *11 MONS 056217 SUBCOMMITTEE OM THE HEALTH EFFECTS OP POLYCHLORINATED BIPHENYLS AND POLYBROMINATED BIPHENYLS Chairperson: A. C. Kolbya, Jr., M.D. Subgroups and Thalr Manbara 1. Chemistry J. Burk* D. Firestone T. Kopp J, McKinney I. Pomerants, Chairperson J. Roach W. Trotter 2. Mataboltaa and Blochaalcal Toxicity C. Frlaa A. Gardnar L. Garthoff J. Goldataln Y. Ku H. Matthews, Chalrparaon J. Moora 3. Animal ToxlcltVand Carclnoaanaala J. Bucklay L. Fiahbaln G. Flaw L. Knara R. Kimbrough, Chalrparaon U. Marcus S. Shlbko R. Tasks 4. Htaaan Expoaura F. Cordis, Chalrparaon P. Comalluaaan vlll MOMS 056218 C. Jellnek B. Hackley R. Lehman J McLaughlin R. Rhoden R. Shapiro Staff Support H. Geratnar J. Moore R* Shapiro \ lx MONS 056219 Subcommittee on the Health Effects of PCBt end PBBe Purpose: 1. "Assemble, review and Interpret date that assess the health significance of polychlorinated biphenyls." 2. "Formulate recommendscions as to future research needs." Specific Objectives* 1. Chemistry: Assess the impurities that may be found in PCBs and PBBe, the "hardness" of the data giving their potential for accumulating in the food chain. 2* Metabolism and Biochemical Toxicity: Asses* the Metabolism of PCBs and PBBs end the poeelble contri bution of specific iaomerlc homologs to the observed toxicity; evaluate poeelble effect* at the cellular level; end define existing dose-response relationships for ths phenomena. 3. Animal Toxicity and Carclnogsnlclty Asaaaa the short and long term effects. Including carclnogenls, c.f exposu.- to PCBs and PBBs In various animal modal#, their relationship to known human effects, end the establishment of dose-response curvet. x HONS 056220 6. Human Exposure Quantity known human exposure*, accidental pulsed, and long range, chair relationship to possible overt symptomatology, and the establishment of possible dose-responea curves. xi MONS 056221 INTRODUCTION PCBa were reportedly first synthesized in 1881 by Schmidt; and Schulte, but were not available commercially until 1930* (Hubbard, 1964). From 1930 until 1970, when their distribution In the U.S, was voluntarily restricted by Monsanto to cloeed systems, the use of PCBe in a wide variety of industrial applica tions had steadily Increased. 1. Commercial PCBs are complex mixturaa. Arochlor 1254, for example, contains 69 different molecules (Zitko, 1971), which differ In the number and position of chlorine atoms. This is a chemical representation of a PCB molecule. C carbon atom, X * Cl (chlorine atom) or H (hydrogen atom). The baalc parent molecule contains two benzene rings connscted by a carbon-carbon bond. Whan all tha X* and X ara hydrogen, the molecule la called biphenyl. When several of tha hydrogen atoms are replaced by chlorine atoms, the molecule le called a PCB. X and X' ara used to indicate that tha chlorine atone are on different benzene rings. For example, the chemical structure for 2,5,2*,5'-tetrachlorobiphenyl Is: Positional isomers have tlu* same number of chlorine stoma, but they at located at different positions on the ring. 2, 5,2',5*-tetrachloroblphenyl end 3,4,5',4*-tetrachloroblphsnyl (TCB) ara isomers. They are different molecules with differ ent chemical, physical, and biological properties, Tha chemical structure for 3,4,3*,4'-TCB is: HONS 056ZZZ The problem of polychlorinated biphenyls (PCBs) became of national concern in 1971, when several Accidental contamination# of foods were reported. In addition, the extent of the environ mental contamination, and Its persistence, were not precisely known. Subsequently, various regulatory actions were taken by the concerned agencies Involved and, with the cooperation of the only U.S. producer, the situation waa felt to be under control. Then, In 1975, the reported high levels of contamination, with PCBs, of Hudson River fish, refocused national attention on this environmental contaminant. It was soon apparent that the actions and control measures of the early 1970*a had not succeeded In totally reducing, or even substantially alleviating the problems assoclatad with this environmental pollutant. On November 17-19, 1975, a National Conference on Polychlori nated Biphenyls, sponsored by the Environmental Protection Agency In cooperation with the Department of Agriculture, Council on Environmental Quality, Department of Health, Education and Welfare and Department of the Interior, was held in Chicago, Illinois. This confsrance brought together the latest data, and scientific expertise, to consider all aspects of the various problems asso ciated with PCBe, and the meant for their possible resolution. In his Introductory remarks to the Session on Health Effects and Human Exposure, Dr. David P, Rail, Saaslon Chairman, announced the formation of a Subcommittee on PCBs of the DHEW Committee to 2 hons S6ii3 Sw Coordinate Toxicology and Related Program*. The Subcommittee was given the charge to: 1. Assemble, review and Interpret data that assesses the health significance of polychlorinated biphenyls. 2. Formulate recommendations as to future needs. The accidental contamination of animal feeda with polybroroinated biphenyls (PBBs), and consequent human exposures, that occurred In the State of Michigan, haa been approached not only on Its own basis, but in comparative relationship to the PCBs, as well. Therefore, the scope of this Subcommittee was expanded to the Subcommittee on the Heelth Effects of PCBs and PBBs. Members of this Subcommittee came from within the Department of Health, Education and Welfare, aa well as including consultants from other Federal Agencies concerned with this difficult problem. The report which follows repressnts the culmination of this Subcommittee's efforts* 3 HONS 056224 GENERAL SUMMARY AND CONCLUSIONS CHEMISTRY The great complexity of PCB commercial mixtures has provided the difficult task of separating the component* of these mixtures and determining the complete Identity of the compounds. Efforts to define these mixtures have been quite successful over the past eevaral yaara (Hutatnger et al., 1974). Identification of the component chemicals in PCB residues presents another difficult problem. Little progress has been reported in this area. Since Che fate of components of commercial PCB mixtures entering the environment cannot be followed directly, inferencea must be drawn about chemical alteration of chlorinated biphenyl compound# In the environment mainly baaed on laboratory studies. The known resistance of aryl chlorides to chemical oxidation end hydrolysis presumably vaa a major factor In tha promotion of PCBs for industrial uses. Exposure to non-metabollc environmental agents la unllkaly to rasult in significant oxidation or hydrolysis of chlorinated biphenyls. However, since appreciable photoalteratlon of chlorlanted biphenyls hse been observed using either eunlight or sunlight-simulating lamps in the laboratory (Huttlnger et al., 1974) as energy sources, It Is highly probabl# that photochsmlcal changes occur in tha environment. These photocheaileal studies have Indicated that reductlva dechlorination to blphanyla 4 MONS 056225 of lower chlorine content is a predominant alteration route although formation of more polar compounds. Including a chlori nated dlbenzofuran, has also been reported (Hutzinger at al,, 1976; Crosby and Moilanen, 1973). The diverse conditions of the environment make it difficult to predict accurately the end products and rates of photoalteratlon of chlorinated biphenyls. From a comparison of carbon-halogen bond-energies, brominated biphenyls might be expected to be more readily altered by light than chloroblphenyle. But because of the lower volatility and different end uses of bromoblphenyls, they may not be ae readily exposed to light as the chlorinated analogs. A typical PCB residue from fish resembles the Aroclor 1254 mixture more closely than it does other Aroclora (Zltko at al., 1972; Velth, 1975). Considering the major components of Aroclor 1254 (Sissons and Welti, 1971) it appears that penta-, hex*-, and heptachlorinated biphenyls tend to concentrate at this trophic level in the biosphere, it is of considerable Interest, therefore. to carry out comparative accumulation, metabolism and toxicity studies emphasizing these higher chlorinated biphenyls using compounds of known chlorine substitution pattern. Individual chlorinated biphenyls are now quite accessible through synthesis and recent studies have begun to obtain these types of biological data. Studies using a group of five symmetrical hexachloroblphenyl Isomers in chicks (McKinney et si., In press) and in mice (Blocca, 5 HONS 056226 1975) indicated that distinct difference in toxicity ore potoilhle and these differences could be related to structure. The Isomers with chlorine substituents in the 4,4' positions appeared to accumulate more rapidly In adipose tissue, shoved increased acti vity in the liver and greater overall toxicity (McKinney, 1975; McKinney et al., in press)* Comparison of two pentachlorbiphenyls in laying chicks again indicated the one isomer with ^'-substitu tion to have greater toxic effects (Ax and Hansen, 1975; Ax et al., 1976). Structure-activity relationships, particularly for biphenyls known to be major components of commercial PCB mixtures, could be useful In assessing the potential hazard of these compounds. A major part of determining human exposure to PCB residues in foods is evaluating the adequacy of the analytical methodology used and the significance of the data obtained. The methods applicable to PCB determination are complex and to be judged adequate should give acceptably reproducible results in the hands of experienced analysts. The degree of auccass of Inter-laboratory collaborative studies helps measure the adequacy of a method. PCB residues are multicomponent mixtures. Many common chlori nated pesticides are extracted from samples aloug with the PCBe. Procedures for separation of PCB residues from Interfering pesti cides thcrfore bee ie important as a prerequisite to quantitation. Several different quantitation techniques have been used (Hutzinger et al,, 1974). Compartaon of PCB residue data la difficult where standardization of the quantitation procedure is lacking. 6 HONS 05622? It has been shown that recovery of PCB mixtures purposely added to food samples, using commonly applied analytical methodo logy, varies with the average level of chlorination. Recoveries tend to decrease with Increasing chlorine level (Stalling ec al., 1972). Analytical procedures for determination of PCB and PBB (the latter is predominantly a hexabromobiphenyl) residues are essen tially similar. They differ somewhat in cleanup of the extract and In the use of a higher column temperature for gas chromato graphic determination of the PBB residue. [The limit of quantltatlo^ for PCBe is generally 0.2 ppm for individual foods and about 0.05 ppm for Total Diet composites using Food and Drug Administration methodology. For PBB residues, the Halt of quantitation Is approximately 0.05 ppm in fata and 0.01 ppm in non-fatty foods. Interlaboratory studies of analytical methods for PBB determination have not been reported. The chlorinated dlbsnzofurans (Cl-DBFs) hava becoaa a focus of concern as the class of contaminant compounds In commercial PCBe most likely to contribute significantly to the toxicity of the PCB mixture. This concern presumably stems mainly from the demonstrated toxicity of 2,3,7,8-tetrachlorodibsnzo-p-dioxln end the simile Ity in structure between dlbenzo-p-dloxln and dlbenzofuran (NIEK8 Conference, 1973; also sse Appendix of Chemistry Report) and from the little toxicity data that has bean published for Cl-DBFs (Bauer et el., 1961), Recent date indicated 7 WOKS 056228 che toxicity of 2,3,7,8-tetrachlorodlbenzofuran approaches chat of ch analogous dlbenzo-p-dloxln at laaat In chicks and in guinea pigs (Moors at al., 1976). If th chlorinated dlbenzo-p-dloxlns can serve as an exanple, chlorinated dlbentofuran toxicity can be expected to vary with number and position of chlorine atoms on the parent ring system. Relationships between structure and toxicity will emerge as more Cl-DBFs are synthesized and tested for toxicity. A range of Cl-DBFs (from dlchloro through hexschloro) have been reported In various Aroclors (Roach and Pomerantz, 1974b; Bowes et al., 19?Sa) and two specific compounds, the 2,3,7,8tetrachloro- and 2,3,4,7,8-pencachloro-dibenzofurane, have been Identified (Bowes et al., 1975b). Quantitation of Cl-DBF contami nants in FCB mixtures, difficult procedure, suggests total Cl-DBF levels In the low psrts per million range (Nagayama et al., 1973; Bowes et al., 1973a). The presence of Cl-DBF In a synthesized. Individual, symmetrical chlorinated biphenyl has been reported (Moron et al., 1973). Chemical analysis of these Individual chloroblphenyls may be necessary prior to their use In toxicological and other biological studies. There have been no published positive findings of Cl-DBFe In environmental et pies or In foods other then rice oil. The finding of Cl-DBFe In such samples would not necessarily Implicate coamerclal PCBs as the source of these contaminants since Cl-DBFs (generally the higher chlorine levels) have been reported In other 8 ONS 056229 Industrial chemicala (Flreatona at al., 1972; Schwatz at al., 1974; Villanueva at al., 1974). It la also possible that Cl-DBFa nay ba foread or altered In the environment. METABOLISM MID BIOCHEMICAL TOXICITY The affect of PCBa and PBB on the hepatic alxed-functlon oxidase (MFO) enaynas has been tha most thoroughly studied of any blochanlcsl paranater that they are known to alter. On a molar basis, tha PBBs are approxlnately five-fold more potent than the PCBs in Inducing Increased levels of tha MPO enzymes (Farber and Baker, 1974). Among the PCBs It appears as if their potency Increases with Increasing chlorination and chlorlna substltutlon In tha para > ortho> neta positions respectively I(Echoblchon, 1975). However, this Induction of the MPO enxymee Is not unique to these compounds and the Induction observed la well within the range observed with many other xenoblotlcs. Tha PCBs are somewhat unique as MFO Inducers In that they Induce the formation of both Type I and Type II P-4S0 (Alvares at el., 1973), but this Induction may have been due to the fact that the conmerclal formulation used in the study was a mixture of twenty or more PCBe which ware metabolised to an even greater number of metabolites. In addition, many. If not moat co--terclal PCB formulation contain traca amounts of chlorinated dlbencofurane (Arakl, 1974). These compounds may bs up to 170 times more potent as MFO inducers than the PCBs. The chlorinated dlbensofuran concentration of Amarclcan 9 HONS 056230 PCD formulations la usually quits low whan they at* produced, howavar, tha effect of long-tera la*unknown. In any caaa, alnca Induction of tha MFO enzymes nay raault In lncraaaad horaona metabolism or carcinogen activation, exposure to tha PCBa and PBBs should be Halted on that basis alone. PCBs and PBBs administered at relatively high concentrations, usually 50 ppm or higher In the diet, to laboratory anlaale have been shown to cause prophyrla (Goldstein at al., 1975), disfunction of the thyroid (Bastoaaky, 1974), Inhibition of various entymee (Pardlnl, 1971), changes In the liver to body weight ratios (Certhoff at al., 1975) various disorders of the liver (Kimbrough st al., 1975), and to alter the level or utilisation of corticosteroids (Wasseraan st al., 1973) and vitamins A, D and S (Cecil at al., 1973; Wong at al., 1974; Combs st al., 1975). Certain of tha lass chlorinated PCBs have also been shown to have a mild estrogenic effect when tested In the lmsMtura rat and nouae (Xlhlstrom, 1973; Nelson, 1974). However, moat of these parameters have not bean studied In sensitive epedea or demonstrated at low exposure levels in laboratory animals. The available data Imply that the PCBa and PBBs containing six or fewer halogen atoms are readily absorbed from the gut of higher animals. The available data alao imply that the PCBs are not excreted to an appreciable extant prior to metabolism to more poler compounds, and that long term PCB storage Is la the skin and adipose tissue (Matthews and Anderson, 1975s). Studies of PB1 10 HONS 056231 metabolism are not yet available. Since tleeue samplea from birds end aannle with known exposures to PCBs usually contain only those PCBs with five or more chlorine atoms, It le assumed that the leas chlorinated PCBs have been metabolised and excreted. On the other hand there la little evidence chat fish can metabolite any PCB and an analysis of fish tissue usually shows a PCB pattern very similar to that to which the fish were exposed (Stalling and Mayer, 1972). Laboratory studies have demonstrated that the rate of PCS metabolism and thus excretion le approximately Inversely propor tional to tha degree of PCB chlorination so long as there are two adjacent unaubstltuted carbon atoms on the biphenyl molecule. When two adjacent unaubstltuted carbon atoms are not present the biological half-life of the given PCB may be a matter of years and accumulation of high tissue concentrations with continued exposure la Inevitable (Matthews and Anderson, 1973a). It should be noted that the major constituent of Plramaatsr BP-6 does not have two adjacent unsubstituted carbon atoms and that the corre sponding PCB has been shown to have an extremely long half-life In tha laboratory rat and probably the huuan population as well. It should also ba noted that metabolism can also result In further complications of the PCB problem because those PCBs which are most readily metabolized and excreted are thoae which are moat likely to be metabolized via arena oxide Intermediates. And where aa the reactivity of arena oxides varies grastly and often 11 KOAlS 056232 ultimately determines the toxicity, autegenlclty or carcinogenicity of the parent compound (Jerina and Daly, 1974), the PCBs offer such a range of degree and position of substitution that It would not be unlikely to find that one or more of these FOB metabolites would have the proper stability to be a mutagen or a carcinogen. Thus a move from the more highly chlorinated PCB formulations to the less chlorinated ones may help solve the long-term residue problem only to intensify other problems. ANIMAL TOXICOLOCV Gleaning the Information that Is now available on animal toxicology makes It obvious that different commercial mixtures of PCBs elaclt different toxic responses In animals, and that different animal epecles vary In their susceptibility to the toxic effects of PCBs. Reproduction la severely affected In mink at a dietary level of 5 ppm Aroclor 1254, and a slight effect Is still noted at a dietary level of 1 ppm (Ringer et el., 1972). In rhesus monkeys reproduction was reduced at a dietary level of 2.9 ppm of Aroclor 1248 (Allen 1975). In rats a dietary level of 20 ppm Aroclor 1254 depressed reproduction while In the same rat strain (Sherman), In a study conducted simultaneously In the same laboratory, a dietary level of 500 ppm Aroclor 1260 was necessary to reduce reproduction (Linder at al., 1974), In comparative studies done with European products, Phenoclor DB6 and Clophen A60, end the American product, Aroclor 12(0, the 12 MONS 056233 European product* war* aora toxic to chicken* than th* American product (Voa 4 Koenan, 1970). Tha difference In toxicity In thla etudy waa attributed to contaalnatlon of th* Kuropaan product! with chlorinated dlbaniofurana and parhapa chlorinated naphthalan**. However, If th* difference* In th* effect on reproduction by th* different Aroclora are coopered, th* contaalnatlon with chlorinated dlbaniofurana nay not b* th* declelv* factor. Hepatic porphyria haa been produced In a number of apaclea, nanely th* chicken, rabbit, Japan*** quail and rat, by a number of commercial mixture*, auch a* Clophen AAO, Fhenoclor UFA, Arodor 1016, 1242, 1234, and 1260 (To* and Koenan, 1970, To* and Been*, 1971, Xvaraon at al., 1973, Ooldateln at al., 1974, 1973). It (hepatic porphyria) ha* not been reported la th* monkey, mink or human (a** Tueho) mammalian apaclea quit* auacaptlbl* to th* toxic effect* of FCB* In other way*. Aroclor 1234 and Arodor 1242 produced hepatic porphyria In tha female rat at do*** lower than Aroclor 1014. Again, different commercial mixture* produce thl* toxic effect at different doa*|* lavela, Th* hepatic porphyria occur* concomitantly with an ineraaa* In ALA aynthataae in th* liver. Mixed function oxide*** are alao induced In th* liver and comparative atudlea with FCB laomer* have augieatad that If the 4,4' poaltlon* on tha biphenyl ring are occupied by chlorine atone tha affect 1* moat pronounced (tcoblchon and Conaau, 1973). The liver la th* prlnary target organ for FCi* In th* rat. larly change* Include hepatomegaly with a concomitant lncraaa* 1} MOMS 056234 In lacottl tnd^lMafc'r*tltllua, lipid acdnaulatlon at highef df*Mr* 'lrtdS*l<?20 j(p for^Aifoeldr 11254'-ad IPWP^gnd dPtia-WiTOll atruahffiJ'ShiWgiW^McKrtila <*ttMHb6drtt24MU>im MVaKkilH Wflll)il#frbM#lirta WfUMc bfftdfWai MfloJ^nMP of&alt4aetUVbf-t*HiLllgti*igi(l4tniu*fmtilittH.0i ddcaMyv bjMaftUgMdadgdttaiWdtM^MtftWa dttd <**U<MaMirdr<Wof((cfMdii> noted with either highl*ade#vdiP#ig#i,Je*flod#i'e'<iMw|*7n97>). These changea are not aa pronounced with Aroclor 1242 or 1016. ,a9lsqa lo ledeiun c nl Bsoubinq upjd ed eirydqioq plieqaH Slnllar changes have been reported In the prlaate (Allen, 197S). isdaun a \d ,2a1 bna Xlaop ecsnsqsl. .Jtddai .nerfotdo erfi xlsabii in<m aiep*i!v9itvMto<$Sv*'*iei/>t>wmi!is*0ii*tmmf > 1256 fltfwhWSih t&na&Ur M6*<iftPrft 1973^Wd .i(t## <WV^^Cat(iMorId^*(r?#vifnd^a,'W?J) b.u 'm4'vtroi?Ah"i2to>'<atatne8A>tt%M4 a&Ut 1242, >42MAitf)r^MiWiMdMr itm? aa -mpitMnfom *<!*Mr0tW<<W<kW*afofrMWHWMry leM*<Wf) tMWMf# 4#ln7aM>3tl<WmlliMl*#6MAi itf oU*#g*rMv#*A*ttf Mpa>*dWPltlM<*tl]7'lMM^ rtgW but mh waryeMgtd >6a Mtgxdupwuwgplita'WildMlMitedlMrvWd^MW^MWssu polluted'ValMlgwa^a '^uto7adWdiOaVpeittli^<*lUMMlai'v4Ma -v) l ewptUaMdW 6y2rltw^gC0vtldin**a4nil4tAyJleva^h*WtV>^d*S4<\ud><w an'WW'i((UK<ldl^tMRAP d!IMynd^^dK,%ffbdV,6Wr^c'eMi*'13 (2.(J%(wdrtvta> PCI alxtures do not raprasant a no affect level. It la presently Jar ndj nl agOI lot negro legrai yrawliq art) tJ vavlx edT not known what the no efface lavala ara. Other uncertainties vnanpni inaiiooonoo a rtllw qlegeeiolsqeri ebulonl Kvgnsrfo yil ^3 Cl 16 HONS 056235 contribute to these problems* The Yuiho oil which caused a poison ing outbreak In Japan was not only contaminated with PCBe but also with high levels of chlorinated dlbenzofurana* 1 In the primate, In addition to the effect on the liver, the gastric mucosa (Allen and Borback, 1973), the skin, and the Meibo mian glands are also affected at comparatively low dietary levels and the bone marrow Is depressed (Allen 1975) while the gastric mucosa of the rat Is only affected at exceedingly high doses (Kimbrough, 1974). Whether the dog also shows an affect on the gastric mucosa needs clarification* In the rabbit, atrophy of the thymus Is a toxic manifestation in addition to liver pathology (Vos and Beams, 1971), Fluid accumulation occurs In primates, chickens, and finches. The lymphatic system Is also affected In minks. The few studies conducted with some hexachloroblphenyl Isomers demonstrated that the 3,4,5,3',4',5' hexachloroblphenyl was the most toxic while 2,3,6,2',3',6' hexachloroblphenyl was the least toxic Isomer (Biocca et el., 1975). Penta-, hexa- and heptachloroblphenyle are preferentially retained In mammalian adipose tissue for extremely long periods of time (Kimbrough, 1975) (at fairly high concentrations In the rat for a recovery period of 16 months). Whether this has an Influence on the toxicity of PCB mixtures has not been determined. The toxic effects of the contaminants of PCBe have not been extensively studied. While it Is assumed thet chlorinated 15 HONS 056236 naphthalenes ir toxic within th* iiu do Mg* rang* as th* chorinatcd biphenyls, th* chlorinated dlbensofurans probably hava a greater toxicity. It 1* assuned that Z, 3,7,8 tetrechlorodlbensofuran la th* nose toxic of this group of compound*. This coapound also shows narked spaclaa variation. While th* single oral LDjo In guinea plga la between 5 and 10 ug/kg bodywelght, 1000 ug/kg TCDP given orally had no affect on rata. Mica are equally Insen sitive to th* toxic effect* of TCDF (Moore at al., 1976). For the bronlnated biphenyl* United toxicity data *r* only available on mixtures containing predoainately hex*- and octabronoblphenyl. Both nixtura* differ sufficiently In lsonarlc composition so that thalr toxic effect* nay quantitatively be quit* different and also different fron tha PCBa. Again th* problan of toxic contaminants ha* not been resolved. If toxic effects are similar to PCBa then, at least In son* specie* such e* mink and monkey, longterm low level exposure should result In neasuraabla toxicity. Additional anlnal studies are needed to rasolv* soma of thasa problems. Poor metabolism and excretion of the brominated blphanyla may load to long ratantloo of thasa compounds predominantly In adlpoaa tissue with accumulation to very high levels on continued exposure. Whether this would lead to sufficient raclrculatlon of th* chemicals to cause toxic effects on target organs In presently not known. 16 MONS 056231 HUMAN EXPOSURE Several reports (Jellnek & Cornelluaaen, 1975; Humphrey, 1976; Kuta and Yang, 1975; Dannla, 1975; Klelnhart, 1975; Heaae, 1975; Yobs, 1972; Kutz and Straeamman, 1975) provide evidence Chat would indicate that a oubetantlal proportion of the population of the United State* haa been exposed to PCB's. Minimal human exposure of the population to PCB's haa occurred from food, air and water, while significant human exposure appears to be limited to sports fishermen consuming fresh water fish from contaminated streams and lakes, and to occupational exposure In industrial workers. Jellnek 6 Cornelluaaen (1975), In reviewing data from the FDA Total Diet Study (1971-1975) report that all food claaaaa of the total diet have declined to no PCB occurrences except In these meat-flab-poultry composites. About 60 percent of these composite* continue to contain detectable residue* of PCB's, although only traces have been detected In the latter years. The fact that levels In these composites havs declined to only traces further support the Inference that tha meat, poultry and eggs no longer contain detectable PCB's and that the low level findings are probably dua to the flah in thaaa composite*. This would Imply that the PCB levels in the diet may hava "bottomed out" and may remain static until such time as thsrs Is a change In the PCB residues In fish. 17 HONS 056238 Data coop Had from atudiea sponsored by tha National Marina Fisheries Sarvlca (NMF5) alao support tha continuing praaanca of PCB raalduaa in fish. A compilation of PCB data, rapraaanting tha results of all tha measurements known to NMFS on PCB'* in fish usad in tha U.S. diet, indicates savaral important points: (1) while at one time or another, soma PCB measurements have been made on many flah, there la an Inadequacy of information on PCB residues in the moat Important fish items in the fish diet; (2) sampling and analyals have been sporadic and not designed to measure trends in human exposure; and (3) systematic surveys of neither the important items nor the species most likely to be contaminated have been undertaken. However, these survey data do show that in general U.S. fish eatera Include a wide variety of fish Items In their diet and that soma 93 percent of the U.S. population (197 million) consume fish, with the average annual consumption of fish balng IS pounds/year/fish eatar. At present. It is difficult to estimate all human expoaure to PCB from eating fish, either from the population as a whole or subgroups at higher risk of consuming large quantities of fish with higher PCI residues. Fragmentary evidence from NKFS data suggests that tha exposure of the population as a whole from PCB reeldues In Ingested fish la probably well below 19 meg/day/ consumer, based on PCB levels which are estimated to be below 1 ppm and 19 g of fish consumad/dey. This can be compared to 16 HONS 056239 the results of the FDA Total Diet Study where It has been estimated that the overall PCB dally intake le on the order of 5-10 ocg/day for the general population. The lover FDA estimate Is based on the methodology of the Market Basket Survey where fish are pur chased at the consumer level and would not be applicable to diets which Include a high consumption of fish from certain areas with high PCB residues. A recently completed study (Humphrey et al., 1976) has attempted to assess some of the consequences of human exposure to PCBs from the high consumption of fish from contaminated areas. The results of this study show that a group of sporte fishermen consumed an average of 24-25 pounds of flsh/person/ysar, with the highest Individual exposure for a two year period reported as 180 pounds per year. PCB residue# in cooked fish ranged from 0,36 to 5.38 pp Although there was a vide range of blood PCB levels for esch quantity of fish consumed, there was a highly significant correlation between the reported quantity of take Michigan fleh consumed end the concentration of PCB In the blood of study participants, with the higher reported fleh consumption being associated with higher PCB blood levela. The blood values ranged from a low of 0.007 ppm in the control group (fish consump tion less then 6 pounds/yeer) to e high of 0.366 ppm in the exposed group (fleh consumption 24-25 pounds/year). These Investigators calculated that the amount of PCB Ingested by the exposed group could average 46,5 mg/yeer end ranged from 19 HONS 056240 14,17 to 114,31 mg/year. While no systematic adverse health effects could be demonstrated in the exposed group when compared to controls, the Investigators caution that any long-term chronic effects are unknown at the present time. Additionally, It can be concluded that exposure similar to those reported In this special group will continue and there Is the likelihood that as sportsfiahlng becomes more popular, larger numbers of people may be exposed in a similar way. Although human exposure to PCBs from air and water is probably minimal, there seems little question that such exposure does occur. Samples of ambient sir collected in Florida, Missi ssippi and Colorado, show that PCBs were present at all locations. The average concentration at each of the three locations was approximately 100 nanograms per cubic meter of air. Studies of surface water from the major drainage basins of ths United States report the widespread occurrence of PCBs In both surface water and bottom sediments. Mean residue levels of PCBs In the surface water ranged from 0.01 to 0.05 mcg/Llter, with a maximum residue level of 20.0 mcg/llter. In Wisconsin, effluents from cooling water In aluminum foundries contained PCBs ranging from 11.5 to 335 ppb* Effluents from paper mills ringed from 0.01 to 25 ppb. Analysis of snow melt water from Wisconsin cities showed PCB residue levels of 0.17 to 0.24 ppb, suggesting that fallout of PCBs from the air may be an Important source of PCBs entering the waters of the state. 20 M0NS 056241 In Michigan! testing of $00 samples of Industrial affluents showed 22 percent had FCB residues greater than 0.5 mcg/llter, 8 percent greater than 1.0 mcg/llter, 6 percent greater than 10 ncg/ liter and 2 percent greater than 100 mcg/llter. With sludge dis posal taking place by Incineration, landfill and crop or pasture application, the continuation of FCBs In the environment teens obvious. Adverse human health effects resulting from FCB exposure have cone primarily from studies of occupational exposure and from human exposure through the ingestion of contaminated rice oil in Japan. Schwarts (1936) provided eome of the earliest reports of adverse health affects due to occupational exposure in the U.S., in which he described skin lesions and symptoms of systematic poisoning among workers who were reported to have Inhaled chlorodlphenyls. There have been numerous reports over the ensuing years describing cutaneous eruptions and of systematic manifestations as well, among marine electricians, machinists, capacitor and transformer manufacturing workers, and others occupationally exposed to FCBe. The skin lesions described by Schwarts In 1936 have coma to ba designated as "chloracne." Fart of the chloracne lesion resembles adolescent acne, but Is generally mors severe and the lesion distribution la Inconsistent with adolescent sens. Hare (1969) and Hasegawa at al., (1973) have reported dermato logic ailments which Include "brown chromodermatosis" of the 21 HONS 056242 dorsal joints of the hands and purple-like eruptions of the face and neck* However, Hasegawa et al., (1973) performed a health survey nt worker* In carbonless copy paper factories, two years after the use of I'Cb in such processes had ceased, and reported no dermal effects nor liver function, urine or blood test abnor malities. PCE blood levels were reported as 0.01 - 0.02 ppm. Typical clinical findings In the human exposure to PCB which occured in Japan In 1968, and which resulted from the Ingestion of rice oil contaminated with Kanechlor 400 Included chloracne and Increased pigmentation of the akin, Increased eye discharge, transient visual disturbance#, feeling of weakness, numbness in limbs and soma disturbance In liver function. Adult Yusho patients had protracted clinical disease with a slow regression of symptoms and signs. In the dose-response epidemiologic study, the average cumulative intake of PCB's leading to overt symptoms was 2,000 mg, with tha lowest dose leading to overt symptoms being 500 mg. However,' Xuratsune et al., (1975) have introduced a new factor Into the Yusho Incidence with the finding that tha rice oil contained chlorinated dlbenaofurans (Cl-DBF) at 5 ppm. Nageyama et al., (1975) reports that tha toxicity of (Cl-DBF) Is said to be from 200 to 500 times that of PCB. Whether or not the (Cl-DBF) con taminant is tha crucial toxic subetance producing the symptoms observed in the Yusho incident, or whether the exposure to the high levels of Kanechlor 400 produced the observed effects, or 22 HONS 0562**3 whether there wee n inter-arttve process taking place ta unknown. i I The data neceseary for determining reasonably accurate time and doee exposure to FIB In Individuals In Michigan la either unavailable or non-exlatent. Attempts to secure sccurate dietary Intake with the FBB levels in food and the duration of consump tion have been unsuccessful. It is hoped that data to be received from the Michigan Department of Public Mealth may provide some basis for crude estimates. While there appears to be no evidence at the moment to Indi cate acute health effecte from exposure to FBB, any chronic effects remain largely unknown. A large seals epidemiological atudy Is expected to get underway shortly to Identify all the farm family members from quarantined farms} a large group of study subjects secondarily exposed to FBB through the purchase of farm products on a regular basis from quarantined farms and a control group of individuals not exposed to PBB contamination. This study will continue efforts to identify eny acute or chronic effecte of FBB exposure through physical examination, biochemistry tests, and dietary histories. Efforts will continue to assess the original Flrsmaater BF-i for the presence. If any, of chemical contaminants which might present human health harards. In the meantime, a ten fold safety factor for FBBs when compared to PCBs appears both reasonable and acceptable based on all currently available scientific data. 23 MONS 0562^ Much work remains to be done concerning the toxicity of PCBs and PBBs and the association of these compounds with any demon strable adverse human health effects. To accomplish some of these needs a list of recommendations follows. 24 MONS 056245 GENERAL RECOMMENDATIONS More complete and additional metabolism etudlea with varloue Individual PCBe and PBBa ahould be undertaken. Attention to the formation of possible toxic metabolites of theae coapounde ahould be considered, aa well aa detecting and quantifying any specific biochemical parameters which may be affected aa a result of such exposures. Baaed on the above, lifetime feeding studies ahould be conducted with selected cosnerclal PCBa and PBBa mixtures, as well aa selected individual PCBa and PBBa. Why some species seea to be nore susceptible to the toxic effects of PCBs, and which are most closely related to man In their response should be determined. In addition, more experi mental data Is needed for the occupatlonel PCBe settings; l.e., via the respiratory or dermal route. Additional toxicological evaluations should be made of flah and other foods which contain high levels of environmentally accumulated PCBs and other con taminants. Appropriate toxicological studies should be undertaken with PBBa In order to be able to make some predictions on possible human effects. The quantitative evaluation of halogenated biphenyl exposure to man with reapect to blood and body fat levels, and any possible health affecta, needs further study. With reapect to PCBa, this may be accomplished by Identifying a population of Individuals consuming large amounts of flah contaminated with high levels of 25 HONS 056246 PCB*. In addition, thoso capped (lultuii I'tatly should he* luchuUni In these efforts. With respect to PBBs, appropriate studies should be initiated with Michigan fern families exposed through consuap* tlon of contaminated meat and dairy products. Individual chemical components, of PCB residues should be qualitatively Identified and procedures for improving quantitation of these residues should be Investigated. Commercial PCB (and PBB) mixtures need to be analyzed further to determine which chlorinated dibenzofurans (or, In the case of PCBs, whether any broalnated dlbenzofurans) are present as contaminants. Specific chlorinated dlbenzofuran compounds must be synthesized for use in development of analytical procedures and to aid in identification of contami nants. Analytical procedures art needed to permit examination of foods for praaenca of chlorinated dlbenzofurans. Toxicological studies should be carried out on chlorinated dlbenzofurans and bromlnated dlbenzofurans. 26 HONS 05624 7 DETAILED RECOMMENDATIONS CHEMISTRY Investigate procedures (such as Usbb and McCall's, J. Chromatog. Scl. 11, 366-373 (1973)) for possible improvement of the quantitation step in the analysis of PCB rssidues. Present procedures for analysis of PCBs yield a "PCB residue" that is cosmonly examined using electron capture GC. Individual components of this residue should ba identified and it should be determined whether any of the electron capture GC peaks can be attributed to Cl-DBFs or Cl-nephthalenes. Analyse commercial PCB mixture (e.g. Aroclors) to determine which Cl-DBFs are present as contaminants. Qualitative end where possible, quantitative studies should ba carried out. Synthesised, individual chloroblphenyle should be examined for presence of chlorinated dlbensofureas before metabolism or toxicity studies are carried out with the biphenyl. (The importance of this la shown by the reported formation of 2,3,7, 8-Clg DBF as a aide-product in the Ullmann coupling reaction used to synthesise the symmetrical 2,2',4,4',3,5'-hexachloroblpheoyl.) Synthesise, purify and characterise by physical and chemical means a range of Cl-DBFs to be used for analytical procedure development as reference standards for confirmation purposes end for suitable toxicological studies. 27 HONS 056268 Develop a suitable procedure for analysis of edible fiah for Cl-DBFs, emphasizing recovery and quantitation of 2,3,7,8-tettachlorodlbenaofuran. Analyte a commercial mixture of brominated biphenyls (Firemaster BP-6) for brominated dlbentofuran content. Analyze long-used heat exchange, transformer and caplcltor flulda for chlorinated dlbenzofurans. Synthesize specific 0j> tritium radiolabeled chlorinated biphenyls that may be needed for toxicity and metabolism studies. Study the reported conversion of certain chlorinated biphenyls to Cl-DBPs under sunlight or sunlight-simulating conditions. If feasible, extend the work to additional Cl-blphenyle. Simulating conditions applicable to heat exchange units and/or transformers, determine whether Cl-DBF content of PCB mixtures Increases when the PCBs are heated and/or exposed to air. Carry out laboratory atudles, simulating environmental conditions, to determine alteration products from Cl-DBFs whan Irradiated or heated. METABOLISM AND BIOCHEMICAL TOXICITY Long-term feeding studies approaching life-time ahould be conducted with the less chlorinated PCB formulations. These 28 MGNS 056249 studies should include at least two of the leas chlorinated PCS fonmlatlons and a series of carefully selected Individual PCBe. Further and more couplets metabolism studies should be done with the more highly chlorinated FCBs, for comparison with those done with FCBs having five or less chlorine atoas. The effect of chlorine position on PCB metabolism and arene oxide formation should be further elucidated. The formation of specific chlorinated dlbeneofurans as possible PCB metabolites should be studied, their blochealcal toxicity in mamaslien species determined and the magnitude of their toxicity ascertained. These studies will also Include those specific chlorinated dlbenxofurana identified In new and used FCBs. If possible, arena oxlda metabolites of several FCBs should be synthesised and their toxicology Investigated. Within an lsoaarlc aeries of FCBs, (s.g., tatra or hexa), metabolism of the more toxic members should ba coaparad to those which are lass toxic. Matebollam of Individual PBBs, comparable to those for FCBs, should be studied. These dete should be related to known toxicltlea. 29 HOMS 056250 The toxicology of known )YH mot.ibol lu\s should ho studied. Porphyrin excretion by the people and aninula exposed in the Michigan PBB incident should be checked. People who are suspected Co have received high expoeurea of PCBa or PBBs should be checked fot any changes In enzyme levels-- e.g., protein-bound Iodine, antipyrene metabolism, serum cholesterol, etc The adsorption, distribution and excretion studies of helogenated dlbenzofurans and PBBs should be studied. Further studies of possible PCB metabolism by fish. Study the degree of chlorination and length of exposure on the estrogenic effects and reproductive organs with particular emphasis on the lover chlorinated PCBa (e.g*, 1016). ANIMAL TOXICOLOGY Inhalation studies with "heated" PCB mixture# slantlacing occupational exposure and dermal toxicity studies should be conducted. Studies as to why some species seem to be more susceptible to the toxic effects of PCBa than others and of theso species, which are moat closely related In their response to humans should be undertaken. 30 MONS 056251 Subacute and chronic toxicity studies ahould b conducted with hexabromobiphenyl (Firemaster FF-1) in order to be able to mice some predictions on human PBB toxicity* The toxicity of fish containing high levels of PCBa which have been accumulated by environmental exposure should be evaluated. It should be established whether and which toxic impurities are present in used PCBs, such as tranaformar oil and capacitor fluid. Long-term feeding studies, approaching life-time, should be conducted with laboratory animals and selected comtrclel mixtures of PCBs and PBBs as well as selected individual PCBs and PBBs. Further and more complete metabolism studies should bs done with individual PCBs and PBBs. The formation of toxic metabolites of these compounds should be considered end cerefully investigated. Efforts to detect and quantitate specific biochemical mechanisms which are affected or altered by exposure to PCBs or PBBe should be contInusd. HUMAN EXPOSERS Polychlorinated biphenyls Recommend that a population of fish esters be Identified who are consuming eubetanclal amount# of fresh wster fish with high levels of PCBs. This study population should bs followed proepectlvely with adequate dietary histories, blood and body fat lsvsls 31 MDNS 056252 of PCBs,* health questionnaire, liver function tests and other biochemical studies. Recommend a similar study be carried out in an industrial population exposed to PCBs. Systematic studies to identify sub-groups of the total U.S. population who consume high levels of fish with more accurate measures of PCB exposure. Polybromlnated biphenyls Recomend that a large scale epidemiological etudy be carried out to (1) identify all farm family members from quarantined farms In Michigan, (2) Identify those lndlvlduele with secondary exposure to PBB contamination through the purchase of dairy producta from quarantined farms on a regular basis, and (3) a suitable control group of non-exposed farm families. These groups should be followed to assess any short and long term adverse health effects related to PBB exposure. Subgroups of these farm families from quarantined farms should be studied by: 1. Repeat PBB blood levels. 2. Matched blood-body fat PBB levals. ' 3. Pregnant femala-lnfant studies. 4. A battery of biochemical studies. 32 MONS 056253 Ancillary Studies Toxicology studies In non~human primates and rodents with hexabromlnated biphenyl and any broolnated contaminants of hexabrotnlnated biphenyls. Toxicology studies In several animal species (nice, rats, dogs, non-human primates using hexabromlnated biphenyl and several PCBs to determine species differences In response to the effects of these compounds. Toxicology studies feeding hexabromlnated biphenyls and selected PCBs singly and in combination to determine if there are different or additive effects when fad in combination. 3) MQNS 056254 CHEM1STKY OF PCBs AND PBBs I . Introduction A recent book by Kutzinger et al. (1974) reviews the chemis try of chlorinated biphenyls through the end of 1973. This wide-ranging review also Includes discussions of metabolism and determination of chlorinated biphenyls. The present report reviews areas of chloroblphenyl and brontobiphenyl chemistry held to be pertinent to the Subcommittee's concern for possible health effects of these types of compounds. No attempt at complete coverage of these subject areas is claimed. Findings reported since 1973 are particularly stressed. Non-metabolic alteration of the halogenated biphenyls is considered in terms of agents most available to Induce chemical change in the environment. Expected differences between chlorinated biphenyls and brominated biphenyls in their environ mental chemistry are commented on. It is possible that certain of the chlorinated biphenyl compounds will be shown to have relatively greater potential hazard for health. Therefore, occurrence and fate in the environment and menbolism and toxic effects in animate are rotated to chemical structure of thu biphenyls. For the general public, food is presumed to be the major V. HONS 056255 Vv' source of chlorinated biphenyl compounds. The important concepts related to analytical methodology for PCB (and PBB) residues, particularly in foods, are reviewed. Alternative means of quantitation of these residues and problems in the quantitation techniques are discussed. The pertinent aspects of chlorinated dibenzofurans, as known contaminants in commercial PCBs and as potentially significant environmental contaminants, are reviewed. In reviewing the chemistry related to health effects of the PCBs and PBBs, it oust be remembered that environmental contami nation by these two commercial chemical mixtures la vastly different in magnitude* The former have bean steadily released Into the environment, in many countries, presumably over decades, and are now found to be a pervasive, world-vlde contaminant. The number of chlorinated biphenyls reaching the environment probably number nearly 100 different compounds. The PBBs, encompassing a small number of chemical structures to begin with, are of concern due to a single, fairly recent contamination incident, apparently limited to the State of Michigan. U. Chemistry of Chlorinated Biphenyl* A. synthesis and analysis of Coimserclal Mixtures Commercial preparation of mixtures of chlorinated biphenyls by reaction of biphenyl with chlorine hae been described by ^ Hubbard (1964). A more recent discussion of this subject, 35 MOMS 056256 including the newer preparation designated as Aroclor 1016, is now available (Mieure ct al., 1976). Analysis of the chlorinated biphenyl content of various American commercial PCB mixtures (Aroclor) has been reviewed by Hut2inger et al. (1974). An estimated 4p to 60 different chlorinated biphenyl compounds are present in each of the higher chlorinated comnwrciul mixture. (There are 209 possible compounds obtainable by substituting chlorine for hydrogen on from one to ten different positions on the biphenyl ring system; see Appendix.) PCB commercial mixtures produced in the U.S. and elsewhere have been shown to contain classes of compounds other than the chlorinated biphenyls: chlorinated napthslenes and chlorinated dlbenzofurans (Cl-DBFs), for example (Vos et al., 1970; Roach and Pomerante, 1974 a,b; Bowes et al., 1975a). The possibility that naphthalene and dlbenzofuran contaminate the technical biphenyl feedstock used in preparation of the commercial FOB mixtures has not been excluded. Available toxicity information indicates that* of the identified types of contaminants in commercial PCB mixtures, Cl-DBFs pose the greatest potential hazard (See Section II.E on Chlorinated Dibenzofurans). Aa a result, the Cl-DBFs have become the focus of studios on contaminants in these mixtures. No report has appeared oi an attempted complete content analysis of the trace impurities in a commercial PCB mixture. 36 HONS 056257 B. Non-aetabollc Alteration of Chlorinated Biphenyls Chlorinated biphenyls, as Is typical of aryl chlorides generally, are quite stable to chemical alteration. Consideration of possible non-metabollc alteration routes for these compounds in the environment suggests air oxidation, aqueous hydrolysis, and photoalteration in sunlight as potential reactions to be Investigated. (Thermal decomposition would be of concern In connection with Incineration conditions meant to destroy waste chlorinated biphenyls). Study of these reactions under the complex and variable sets of conditions existing in the environ ment is difficult. Chemical reactivity of chlorinated biphenyls has therefore been studied almost entirely under the more con trolled conditions of the laboratory. Oxidation PCBe are fairly stable to oxidation under moderate conditions. PCBb art stable under conditions which easily oxidise DDE, and can be separated from PDB by oxidising DDE to the more polar dlchlorobensophenone prior to column chromatographic separation (Trotter, 1975). To effect the oxidation of DDE, the PCB-DDE solution is refluxad in 2.5E chromic acid-acetic acid on a steam bath. Some cf the lov ,r chlorinated biphenyls, however, are not recovered after the oxidation. It is reported (Welngarten, 1961) that mono**, di-, and trlchloroblphenyls are oxidised in 7.5t 17 HONS 056258 chromic acid-acetic acid to their respective (chloro-) benzoic acids. With vigorous oxidizing conditions in the environment* some PCB oxidation, especially of the lower chlorinated biphenyls may occur. It is difficult* however* to assess the total extent of possible environmental oxidation of PCBs. Hydrolysis and Alcoholysis PCBs are fairly stable to hydrolysis under moderate conditions. When refluxed with 21 KOH In ethanol* PCBs are stable (Trotter* 1975; Young and Burke* 1972). A halogen on a chloroblphenyl molecule is not easily displaced in a nucleophilic substitution reaction. Under vigorous conditions the 4 and the 4*4' positions of decachlorobiphenyl are found to be the most susceptible to chlorine displacement by hydroxide and methoxlde ions. Deca chlorobiphenyl can be hydrolysed to octachloro-44*-blphenylol when treated with aqueous alkali at high temperatures In an autoclave (Smith* 1948* Soclete d'Electro Chlmle* 1962). Heating 25-dlchlorobiphenyl with sodium methoxlde produces 2-chloro-5-biphenylol (deCrauw* 1931). Decachlorobiphenyl when treated with sodium methoxlde in pyridine ylelde the 4-aethoxyand 4.4'-dimethoxy-chlorobiphenyl (Blnna end Suachlttky* 1971). It is probable that the ron-metabolic hydrolysis or alcoholysis of PCBs in the cwironmen Is limited. Photochemistry Exact environmental conditions for the photochemistry of 38 HONS 056259 environmental contaminants, Including PCBs, are sometimes difficult to simulate In the laboratory. Limited understanding exists concerning the parallel of laboratory photoreactions and reaction rates under non-envlronmental conditions and possible environmental photolyses. The exact environmental conditions under which residues may photoreact can be diverse and difficult to evaluate. Unknown or discounted factors In the environment, such as possible senslters or quenchers, may be important. The frequency (energy) and Intensity of a laboratory photolysis light may not be comparable with environmental conditions. 3000 A probably represents tht practical lower limit of the ultra violet (UV) portion of sunlight (Crosby, 1969). Th* residue must be exposed to sufficient light to react appreciably. It Is necessary to consider the physical state of the residue exposed to light In the environment. The residue may exist se a solid or liquid (e.g., a film), a solution or a vapor. In photolyslng solutions the solvent can have an iaq>orcant effect. Photoreactions which proceed In one solvent may not proceed or may proceed at a different rate in another solvent. A muddy river containing a residue may yield different results compared to a laboratory photolysis In hexane. Laboratory photolysee under a variety of conditions which simulate moat, if not all, of the environmental conditions of photolysis could be extremely sign ificant and relevant. In the absence of these, we must rely on the questionable extrapolation of laboratory photolyses under non-envlronmental conditions. 39 HONS 056260 The photochemistry of PCBs has been described by Hutzlnger, et el.,(1972; 1974). In contrast to oxidation and hydrolysis, PCBs are fairly easily photoreacted under certain laboratory conditions. PCB photochemistry has been studied in various solutions, as s thin film, and as a vapor. In hydrocarbon solvents progressive reductive dechlorination of the PCB is the predominant photochemi cal reaction (Hutzlnger et al., 1974). Irradiation of solutions of chloroblphenyls (Hutzlnger et al., 1972) and trapped gas liquid chromatographic (GLC) effluents (Hannan, et ai.,1973) have shown that PCBs with higher chlorine content dechlorinate more readily. 0.1X hexane solutions of 2,2 * ,5,5'-tetrachlorobiphenyl and 2,2*,4,4't5,5'-hexachlorobiphenyl were each irradiated with 3100 A light In a Rayonet reactor (Hutzinger et al., 1972). After 24 hours of irradiation, 33X of the 2,2'(5,5'-tetrachloroblphenyl was unreacted while < IX of the 2,2'4,4',5,5'-hexachlotobiphenyl remained. In dechlorlnatlng PCBs by photoexcitation, ortho chlorines preferentially cleave (Ruzo et al., 1974). With PCBs containing only nets and para chlorines, meta chlorines are lost preferentially. The rate of dechlorination is faster in alcohol solvents, such as methanol (Hustert and Korte, 1972) than in hydrocarbon solvents. Irradiation of 2,2',4,4*,5,5'-hexachlorobiphenyl In methanol yields, in addition to reductive dechlorination, *-lng metnoxylatio.. (Ruzo et al., 1974). Photochemical studies of PCBs in aqueous solutions may offer a significant relevance to environmental PCB photochemistry. A 0.4X Aroclor 1254 suspension in vater-dioxane (7+3) In the presence of sodium bicarbonate with 40 HONS 056261 air bubbling through the mixture was irradiated with 3100 A light. Two thin layer chromatographic fractions of the Aroclor 1254 after irradiation corresponding to the "addition of water to chlorobiphenyls" and a "carboxy fraction" were found (Hutilnger et al., 1972). Presumably PCBs formed by reductive dechlorination were also produced. Black light irradiation of Aroclor 1254 as a thin layer film In the presence of water yields a "carboxy fractic. ' and a fraction whose mass spectrum indicates hydroxychloroblphenyls (Hutzinger et al., 1972). Sunlight irradiation of certain chloro biphenyls as a thin film without the presence of water gives reductive dechlorinated products (Hutzinger et al., 1974). Chlorinated terphenyls and quaterphenyls are also produced by the black light and sunlight Irradiation of certain chlorobiphenyls as a thin film (Hutzinger et al., 1974). Vapor phase photolysis of PCBs may be extremely relevant especially for the more volatile components of Aroclors with low chlorine content. Sunlamp Irrad iation of the vapor phase of a refluxing suspension of 2 2*,5,5'tetrachloroblphenyl and water yields "carboxy" products (Hutzinger et al., 1972). These laboratory photoreactions represer. chemical conversions which may proceed In the environment. The photoreaction rates under diverse c vlronmental conditions and the extent of PCB degradation or reaction In the environment are extremely difficult to assess. HONS 05626Z The reported low-yield conversion of certain chlorinated biphenyls to Cl-DBF is discussed under Chlorinated Dibenzofuran, Section II.E. C. Chemical Structure Related to Occurrence, Fate and Effects of Chlorinated Biphenyls the physical and chemical properties of chlorinated biphenyl compounds and commercial mixtures of PCBs vary greatly depending on the degree and position of chlorine substitution on the biphenyl ring system. Of particular Importance to their environmental occurrence and fate are the properties of volatility, water solubility, bloaccumulatlon, biodegrndability and photoatabllity. Volatility, water solubility and bloaccumulatIon are of more importance as mechanisms of introduction Into and transport within the environment. Biodegradation and photodegradation are more Important as mechanisms of removal. There is data (Mieure et al., 197$) which indicates that the higher chlorinated biphenyls are leas volatile and less water soluble than the lower chlorinated biphenyls. These two factors would tend to enhance the ratio of lower to higher chlorinated biphenyls In the environment. There is no clear data to assess the Involvement of chlorine position in volatility itMi water imlubility, bur is likely to be of less importance. Therefore, the lower chlorinated biphenyls arc more likely to volatilize Into and be selectively transported in the aqueous environment. 42 HONS 056263 The relationships between structure and bioaccumulation factors for PCB isomers are only beginning to be assessed (McKinney, X975; Suglura et al, 1975) with the availability of purified specific Isomers for study. However, It appears (Zltko et al., 1972; Vetth, 1975) that the peak patterns from gas chromatography found In environmental samples representing the biosphere most closely resemble the higher chlorinated com mercial mixture, Aroclor 1254. This suggests a selective accumulation of the mores highly chlorinated components of the commercial mixtures in biological material. This also indicate that the higher chlorinated biphenyls are able to find their way into the environment In spite of their poorer volatility and watar solubility. Pharmacokinetic studies (Matthews, 1975) with selected radiolabeled chlorinated biphenyl compounds have confirmed the trend toward increasing biological half-life with increasing chlorine number. However, these findings may be the result of both higher accumulation rate# and lower elimi nation rates for the higher chlorinated biphenyls. It may be of interest to note that fish from lower and intermediate levels of the food wsb have been found (Zltko et al., 1972) to contain lower amounts of hexa and higher amounts of tetra and pants then higher trophic level white and silky sharks and aquatic bird# This suggests that selectivity In bloaccumulatlon is also s function of blospeclv*. Again the effects of varying chlorine position are less 4) HONS 056261! clearly understood, and the best assessment of this comes from study of two or more members of an isomeric series of the chlorinated biphenyls. Recent work with five symmetrical hexachlorobiphonyl isomers in chicks (McKinney et al., in press; Goldstein et al., in press) and subsequently In mice (Blocca, 1975) has demonstrated that separate and distinct differences in isomer toxicity are possible and that the differences are related to chemical structure via effects of varying chlorine substitution on compound llpophlllclty and metabolism. The hexachloro isomers studied were 3,4,5,3' ,4*,5*-; 2,4,6,2',4',6*-; 2,3,4,2\3\4'-; 2,4,5,2\4' ,5'-; 2,3,6 2\3\6'- and 2,3,5 2', V,5*-. Those hexa-lsomers with 4,4'-aubstltutlon appear to be more rapidly accumulated and may be more slowly metabolized, i.e., these isomers showed greater accumulation in adipose tissue and increased activity In the liver and overall greater toxicity. Theae differences are believed to be associated with differences in molecular polarizability and to be measureable by spectroncopic and chromatographic techniques. Other workers (Ax et al., 1976; Ax and Hansen, 1975) have compared the toxicity of two pentachloroblphenyls in laying chicks. The isomer with 4,4''Subititutlon (2,4,5,3*,4*) shoved higher ''orago embryonic mortal Uy and teratogenicity in unhatched eggs but lover decreased fertility in >-he chick than 2,3,6,2*,3* the isomer not chlorinated in the 4-positlon. Although it is difficult to assesa the role of metabolism here, the data would suggest a 44 HONS 056265 more rapid accumulation of the 2,4,5,3'4'-isomer In the egg. Other workers (Bush et el.. 1974) have already obeerved a correlation between PCB content of egge and embryo mortality and teratogenicity. Various other workers (Ecoblchon and Comeau. 1975; Johnstone et al.. 1974; Hill et al. 1974) studying a range of chlorinated biphenyls have generally observed the Importance of degree of chlorination as well aa position of chlorine sub stitution (especially 4,4'-eubstltutlon) In overall biological effectiveness. Although some of tht compounds studied ere unrealistic aa components of comnercial PCB mixtures, they have served as models to demonstrate the importance of chlorine number and position. Nevertheless, of 50 possible components of Aroclor 1254 identlfed by Sissons and Welti (1971) as many as 19 (38Z) could have 44'-substitution. It la of lntarest to note here that one of the possible persistent Isomers found In Yusho patient tissue has bean tentatively identified (Kurateune, 1975) aa the 2.3.4.5,3*.V-haptachlorobiphenyl. This isomer has been prepared (McKinney* personal communication) and Its toxicty will be determined especially in relation to the highly toxic 3.4,5.3*.4*,S'-hex* leomer previously tested. ^ It has bean only recently that hydroxylated metabolites of PCBa have bean Isolated and Identified (Janaeon et al.( 197S) In environmental easiples. The study of the metabolism of the commercial mixtures (deFreitas and Norstrom, 1974) themselves has 45 HONS 056266 received little attention for obvious reasons. Recently, the metabolism of purified chlorinated blphneyls, some radiolabeled for quantitation purposes, has been studied in a number of biological systems. Although there are a large number of publications in the literature on thi9 subject* this report will concern Itself with those studies which seem to support general** Ities In terms of the effects of varying chlorine number and position on metabolism with particular reference to mammalian systems and to realistic components of Aroclors. The distribution and excretion of a series of four radio labeled (1<iC)chlorobiphenyls which have degrees of chlorination similar to and are themselves constituents of Aroclorn 1221, 1242, 1254, and 1260 have been studied (Matthews, 1975) in the male rat. These studies have clearly shown an increasing biological half-life with Increasing chlorine number for the series (4-; 4,4'-; 2,4,5,2',5'- and 2,4,5,2',4',5'-) which Is believed to be related to the rate of metabolism and the ease of formation of the arene oxide Intermediate. These compounds were also dosed at realistic exposure levels (0.06 to 6.0 mg/kg) which showed that little if any of the chloroblphenyi Is excreted in unchanged form. Most other metabolic studies (Hutzlnget et al., 1974) with selected chjorobiphenyls have dealt with much higher dose levels (generally resulting in excretion of much unchanged biphenyl) and unlabeled compounds which are of little quantitative value in assessing the effects of chlorine degree and position. Ah HONS 05626? There has been no similar study with selected chlorinated biphenyls vithln an Isomeric series to assess the effects of chlorine position on metabolism with the possible exception of an Incomplete study (Hass et al., unpublished) on the identifi cation of metabolites from the excreta of chicks (and mice) fed symmetrical hexachloroblphenyl Isomers. It Is essential that this be done with symmetrical Isomers (In order to simplify the probelm of interpretation) and preferably In two or more Isomeric series (tetras and hexas). The various metabolism studies have generally shown the occurrence of polar hydroxylated compounds as major components of the metabolite mixture. As the degree of chlorination Increases, hydroxylatlon can be concurrent or concomitant with dechlorination (Hutslnger et al., 1974). There la also svldsnce for the formation of methyl ethers (Safs^ 1975) and methylchloroblphenyls (Hass et al., unpublished) as metabolites. There Is an increasing body of evidence (Safe et al., 1975) to support the arena oxide intermediate In hydroxylatlon, especially (Chen et el.. Division of Nutrition end Disposition, in press) In cases where vicinal (1-2,-adjacent) unsubstituted carbons ere found In the molecule. The corresponding dlhydrodlol and/or catechol generally occur along with the phenolic metabolites. Althought it has not baen studied in mammalian systems, there Is at least one report (Baxter et al., 1975) that studies of 47 HONS 056268 Individual chlorinated biphenyls do not accurately predict the rates of metabolism of the same compound in simple mixtures. Therefore, the overall problem of metabolic degradation of the commercial mixtures may be complicated by the possibility that certain PCBs are potent enzyme inducers but poor substrates and vice versa with various degrees in between. This is further complicated by the (act that certain chlorinated biphenyls occur in optically active forms and only one enantiomer may be biologically active and undergo enzyme interactions. The existence of nine of the major, and ten of the minor, constituents of Aroclors 1242, 1254, and 1260 In optically active forms has been predicted (Kaiser, 1974). D. Determination of PCB Residues The determination of PCB residues has been reviewed in detail by the WHO Task Croup report on "Environmental Health Criteria for Polychlorinated Biphenyls and Terphenyls" (WHO Task Group, 1975) and in "Chemistry of PCBs" (Hutzinger et al., 1974). PCBs are lipophilic, quite similar In this respect to DDE, the metabolite of DDT. An analyses for PCB residues follow procedures the same as or similar to those used for multiple residues of orga tchlorine pesticides (WHO Task Group, 1975; Hutzinger et al., 1974; Food and Drug Administration, 1968-1975, Horwltz, 1975; Hearing Clerk, Dept. HEW, 1973). The individual steps that follow sampling are extraction of residues from the sample, isolation of residues from coextract Ives (cleanup), 48 HONS 056269 separation of PCBs from Interfering chlorinated hydrocarbon pesticides, quantitation, and confirmation of Identity. In general analytical methods capable of completely extracting residues of organochlorlne pesticides from sample substrates and of quantitatively recovering the pesticides through subsequent cleanup procedures are also capable of achieving quantitative analysis for PCBs. The llpophlllclty of chloroblphenyla which Increases with Increasing chlorine content may Influence their recovery through some analytical methods (Stalling et al., 1972). For example, recoveries through the frequently used cleanup step, partitioning of PCB from a petroleum ether or hexane solution of fat or oil to acetonitrile, exceed 95 percent for Aroclor 1242 but drop to 75-80 percent for Aroclor 1260 (Hearing Clerk, Dept. HEW, 1973). Overall ability of typical analytical methodology to recover PCBs added In vivo to food samples Is shown by two Interlaboratory studies con ducted within the Food and Drug Administration (FDA) and one study conducted for the Association of Official Analytical Chemists (AOAC) (Food and Drug Administration, 1971; Burke, 1972; Food and Drug Administration, 1973; Sawyer, 1973). Using method ology described in the FDA Pesticide Anelytlcel Manual and In the Book of Official Method* of the AOAC, average recoveries and co efficients of variation for determination of Aroclors sddsd es unknowns to dlffersnt foods ware: Aroclor 1254-flah, 74+9 per cent; Aroclor 1254-Infant chicken, 89 + 22 percent; Aroclor 1242chicken fat, 101 + 13 percent; Aroclor 1246-chlcken fet, 96+9 49 MONS 056270 percent;Aroclor 1254-flsh, 75 + 14 percent; Aroclor 1260~fish, 75 + 15 percent. The three studies* involving fish required separation of the PCBs from the DDT group before quantitation. Gas chromatography with electron capture detection is the most widely used procedure for determination of PCB residues. Cae chromatographic columns with methyl silicone liquid phases are widely used under conditions that typically separate the Aroclors and PCB residues Into about 15 peaks (fishbein, 1972). The 10 percent DC-200 (or 0V-101) column described in the FDA Pesticide Analytical Manual separates Aroclor 1254 into 14 peaks (Armour, 1972). These analytical columns do not give a highly defined representation of the Aroclor or PCB residue; several of the peaks in Aroclors result from mixtures of more than one chloroblphenyl (Sissons and Welti, 1971; Stalling and Huckens, 1971; Webb and McCall, 1972; Webb and McCall, 1973). A column prepared from purified Apiezon L has been shown to separate a 54 percent chlorine PCB mixture into over 40 peaks (Jensen and Sundstr&m, 1974). Separation of the PCB residue into three fractions by chromatography on charcoal prior to examination on the Apiezon L column has made it possible to characterize and quantitate nearly 60 technical PCB components. This method provides way to get much needed, more detailed information on the composlton of PCB residues and might be used in special studies. The increased time and complexity of this approach would probably limit Its application in regular monitoring analysis. MONS 056271 The need to confirm residue Identity end the procedures avsllable are similar for PCBs and pesticides. The multipeak gas chromatographic pattern of a PCB residue may be very similar to chat of a commercial Aroclor or, as In most biological samples, the residue peak pattern may be changed to varying degrees from that of a given Aroclor. The use of column chromatographic or chemical reaction procedures to separate PCBs from organochlorlne pesticides Increases the certainty of the gas chromatographic Identification. Procedures readily available to the residua lab oratory for confirming the Identity of PCBs Include: halogen specific gas chromatographic detectors (Hearing Clerk, Dept. HEW, 1973), stability of the residue peak pattern after refluxing the extract with alcoholic alkaline solution (Young and Burks, 1972), perchlorlnatlon of the residue to the decachloroblphenyl derivative (Berg et el., 1971; Armour, 1973), and thin layer chromatography (Fehrlngar and Westfall, 1971; deVos and Pact, 1971). Hass spectrometry Is not readily available to many laboratories conducting analyses for PCBs nor would the expense justify regular use in monitoring programs. However, the use of mass spectrometry Is encouraged for residues and samples which ara unusual or signi ficant and for tha occasional examination of a so-called routine sample. Two especially critical considerations ara associated with the determination of PCB residues: (1) their separation from potentially Interfering organochlorlne pesticides, particularly 51 HONS 056272 DDT, TDE,and J)Dli or the multicomponent chlonlone or toxaphene, and (2) quantitative measurement of the multicomponent PCB residue which, in biological organisms, 19 usually changed in relative amounts of chlorobiphenyl components from commercial Aroclors or which may result from mixture of PCB from different sources. To achieve reliable residue results it is essential that the analyst correctly make critical Judgements and interpretations in dealing with mixed residues of PCBs and organochlorine pesticides and in quantitation of the multicomponent PCB residue. There is no substitute for analyst experience In this analysis. PCBs are separated from certain pesticides in Lhe usual cleanup procedures, e.g., adsorption chromatography on Florlsll or alumina or by gel permeation chromatography (Food and Drug Administration, 1968-1973; Stalling et al. 1972; Holden and Marsden, 1969). When DDT* TDE, DDE, chlordane, or toxaphene are present, ancillary procedures designed to separate these chemicals from PCBa must be used. The DDT analogs, particularly DDE, are the pesticide residues most frequently encountered in samples and DDE is the most difficult to separate from the PCBs. The effective electron capture gas chromatographic response for DDE Is 20-30 times greater than for an equal weight of Aroclor 1254 and the response for DDT is only slightly less than that for DDE (Food and Drug Administration, 1968-1975). Without proper treatment these pesticides can readily interfere in the deter mination of PCBs. Some procedures utilizing column chromatography r>2 HONS 056273 on alumina or Florlsll separate DDT and TD8 and Intentionally collect DDE and PCBS in the same fraction (Holden and Marsden, 1969; Reynolds, 1969), which Is analyzed. In these cases the analyst.must exclude the DDE region of the gas chromatogram from the quantitative measurement of PCBe* Column chromatography on charcoal also has been proposed for separation of PCBe from DDE, DDT, and other organochlorlne pesticides (Berg at al., 1971). Column chromatography on silicic add la probably the most widely used procedure for separating PCBe from DDT and analogs (Food and Drug Administration, 1968-1975; Armour and Burke, 197D). The technique Is fairly lengthy and difficult to reproduce, requiring empirical standardization In each laboratory (Sawyer, 1973; Maaumoto, 1972; Edwards, 1974). The separation of DDE is probably not 100 percent effective but under ideal conditions practically all DDE can be separated; the usual inconsistency la for a portion of the DDE to be eluted from the column with the PCBa, requiring allowances to be made in PCB quantitation. The lower chlorinated PCBa present the greatest difficulty in separation from the DDT group by colutn chromatographic procedures. DDE can also be separated from PCBa by oxidation to the dlchlorobenzophenone, followed by a chromatographic separation of this more polar darlvatlve from PCBa. DDT and TDE may be dehydrochlorlnated to their respective olefine and similarly separated along with DDE (Mulhern at al., 1971; Collins et al., 1972; Trotter, 1974). This technique has not received as much application aa the chroma tographic procedures, probably because pesticides and lover 53 HONS 056274 chlorinated biphenyls are destroyed or changed, preventing their determination. A variation on this approach used sodium dlchrotnate plus a minute amount of sulfuric acid, rather than dehydrochlorlnation followed by oxidation with chromium trloxide in acetic acid. The dlchromate reagent is reported to convert DDE quantitatively to the dichlorobensophenone without affecting DDT, TDE, or any of the chlorobiphenyls (Jensen and Sundstrom, 1974). Quantitation of PCB residues is done in most laboratories by comparison of measurements made on the multicomponent electron capture gas chromatograms of the residue and a known quantity of reference material. The PCB residue in biological samples, as mentioned earlier, Is very likely to be changed In the relative amounts of chlorobiphenyl components from any one Aroclor or the residue may be a mixture of PCBs from different sources (Food and Drug Administration, 1966-1973; Jensen and Sundscrbm, 1974; Cook, 1972). The response of the electron cap ture detector varies with the number and location of chlorine atoms In the biphenyl molecule (Gregory, 1968; Zltko et al., 1971). With the electron capture system described In the FDA Pesticide Analytical Manual, the response/unlt/welght increase* about 6 fold fror Aroclor 1242 to Aroclor 1260 (Hearing Clerk, Dept. HEW, 1973). The halogen specific mlcrocoulometric and electrolytic conductivity detectors respond proportionally to the weight of chlorine present and in theory could provide a more accurate measurement of a PCB residue thnt la not exactly the 34 HONS 056275 same composition as the reference (Hearing Clerk, Dept. HEW, 1973). However, lower sensitivity, difficulty In maintaining optimum performance, and limited availability In residue labor atories has limited the use of these detectors. Unless the PCB residue and reference Aroclors are exactly the same In chloroblphenyl composition, the gas chromatographic determination cannot be considered accurate; the greater the difference between residue and reference composition, the greater the deviation between determined and the true residue level (Hearing Clerk, Dept. HEW, 1973; Beezhoold and Stout, 1973). The most frequently uBed approaches to quantitation have been selected for the reference the Aroclor with the most similar gss chromatographic pattern to the residue and*. (1) compared the response of a single peak In the residue with the response of the counterpart peak in the Aroclor reference; or (2) compared the total response for several peaks from the residue to the total response for tha counterpart peaks In tha reference; or (3) compared the total response for all peaks In the residue with the total response for all peaks in the reference; or (4) In a greater effort to duplicate the residue peek pattern, prepared a reference made up of one or more Aroclors to simulate the gas chromatographic pattern of the residue, and compared the total response for all peaks of the residue to the total response for all peaks In tha reference (Sawyer, 1973; Beethold and Stout, 1973), Response has been measured In terms of both peak height and area (Sawyer, 1973). The latter approach which utilizes an Aroclor or a mixture 55 HONS 0562?6 of Aroclors to duplicate the residue peak pattern is recommended by the AOAC for quantitation of PCB residues in certain foods (Hocvitz, 1975). This is a currently accepted and practical way to quantitate PCB residues. Perchlorinatlon of PCBs to decachlorobiphenyl has been suggested as a means to Improve precision and consistency in PCB determination (Berg et al., 1971; Armour, 1973). This approach will not Improve the accuracy of quantitation, however, because proportions of Individual chloroblphenyls In the PCB residue remain unknown; equal weights of individual chloroblphenyls of different chlorine content result in different weights of decachlorobiphenyl. Contaminations found in antimony pentachlorlde, the perchlorinatlon reagent, also detract from this procedure for quantitation purposes (Trotter and Young, 197S). An approach to quantitation which appears to have practical merit as well as offering improved accuracy in quantitation has been advanced by Webb and McCall (1973). The PCB residue la quantitated peak by peak in comparison to reference Aroclors which have been characterized as to the number of chlorines and the fraction of total Aroclor weight represented by each electron capture peak after separation on a widely used (methyl silicons liquid phase) gas chromatographic column. The availability of carefully characterized Aroclors and evaluation in pructice ;tre required to fully evaluate the merits of this procedure. HONS 0562?? Precision In the lnterlaboratory determination or PCBa la lightly Ie than with the common organochlorlne pesticides. In several Interlaboratory studies Involving biological samples and paperboard containing either added Aroclors or actual residues of PCB the coefficients of variation are about + ZO percent. The results of studies conducted by the AOAC end the FDA Involving samples containing added Aroclors have been mentioned above (Food and Drug Administration, 1971; Burke, 1971,. Food and Drug Administration, 1973; Sawyer, 1973); the recoveries of added Aroclors ranged from 74 to 101 percent and coefficients of variation from 9 to 13 percent. The levels of Aroclors added In these studies ranged from about 2 to 8 ppm with the exception of 0.2 ppm Aroclor 1254 added to chicken infant food. With samples containing actual residues, 9 laboratories In the AOAC study reported 9.2 ppm + 8 percent for a residue In chicken fat and 4.5 ppm + 20 percent for a realdue in Lake Michigan chubs. The residue in the chubs waa determined after separation of DDE, DDT, end TDE by colwu chromatography on silicic acid. In a study by 8 laboratories In cooperation vlth the International Council for the Exploration of the Sea, PCB residues determined In a fish oil varsged 1.97 ppm + 47 percent. A much better coefficient of variation, about + 11 percent, wae obtained when the seme fish oil was fortified with an additional 10 ppm PCB (International Council for Exploratoln of the See, 1974). In an AOAC study of the method for PCB In paperboard 11 laboratories analyzed a paperboard sample manufactured to contain Aroclor 1242 and reported 5.6 ppm + 57 MONS 0562 7e 16 percent (Finstervalder, 1974). The Inter- or Intra-laboratory precision of the determination of PCB in any sample type la improved by use of the same analytical procedures, especially in quantitation of the residue. The lower limit of quantitation for PCB residues will vary among laboratories depending upon the objectives of their analyses and upon the particular details of the analytical methods, especially the sensitivity of gas chromatographic detection and the sample size. The quantitation limit achieved will be higher than for organochlorlne pesticides because of lower effective detector response to the technical PCB mixtures; 20-30 times more Aroclor 1254 or 30-50 times more Aroclor 1242 than DDE Is required for the major peaks to produce the same peak height as DDE in electron capture gas chromatography (Food and Drug Administration, 1966-1975). Other factors which may restrict the attainment of low limits of quantitation and affect analytical reliability are interfering residues In the sample and laboratory contamination. Fish from some locations contain residues of organochlorlne pesticides and possibly other contaminants at levels that would cause Increasing difficulty as the PCB residue decreases below about one ppm. Contamination of laboratory equipment, reage ks, and samples with PCB from containers, previous samples, control runs, und unknown sources, must be carefully guarded against In analyses where a low level of qwantitatlon is necessary, for example with human blood or certain 33 MONS 056279 environmental samples (Trotter, 1975; Jensen et al., 1972; Glam and Wong, 1972). Perchlorlnatlon of the PCB residue to decachlorobiphenyl offers the possibility of about 25 fold Increase in sensitivity in the gas chromatographic determination (Armour, 1973). However, contaminants present In antimony pentachlorlde severely restrict or prohibit application of this procedure to determination of low levels of PCBa (Trotter and Young, 1975). Analytical methods used by the FDA for PCB residues also recover chlorinated napthalenes (Armour and Burka, 1971). If present alone, chlorinated naphthalenes would be recognised by the analyst. However, their presence in admixture with PCBa would probably go unrecognised unless amounts were greater than PCBs or the residue was examined by mass spectrometry. Chlorinated naphthalenes are oxidised by procedures used to differentiate DDE In the presence of PCBe and could be removed from Interfering in the PCB determination (Holmes and Walden, 1972). From examination of procedures used to analyse for chlorinated dibenzofurans In commercial PCB formulations and from analytical studies with chlorinated dlbensodloxlne it t m be Inferred that some chlorinated dibenzofurans may be recovered along with PCBa through anelyt''al methods using Florlsli and column chromato graphy (Vos, et al., 1970; Roach and Pomerants, 1974; Bowes, et al., 1975; Porter and Burke, 1971). The chlorinated dibensofurana have gas chromatographic properties similar to the PCBs (Bowes, 1975). 59 MONS 056280 It is very doubtful, however, that a relatively small amount ol chlorinated dihon/ot uron id the presence of PCBs would ) roro^nlzcd in the usual analysis lor PCBs. Little study has been made of residue analytical methodology for chlorobiphenylols (hydroxylated chlorinated biphenyls). Procedures used for their determination differ substantially from those used for PCBs (Bache and Llsk, 1973; Zltko et al., 1974). It Is not likely that chlorobiphenylols would be recovered through column chromatographic and separation procedures used for PCBs. In the discussion of PCB residues In foods given In the report of the Human Exposure Croup a large share of the Informa tion comes from the FDA's surveillance programs. The analytical methodology used in the analyses is described in the FDA Pesticide Analytical Manual, Vol. I, and in Official Methods of Analysis of the AOAC. Quantitation of PCB residues Is by gas chromatography with electron capture or halogen specific electrochemical detectors. The total response (area or peak height) for ths PCB residue is compered to the total response for the Aroclor reference (or mixture of Aroclors) having the most similar gas chromatographic pattern. Aroclor reference materials are each from a single master lot. Results are reported as ppm of the Aroclor(s) used for the quantitation reference. With certain exceptions ths limit of quantitation for PCBs (based on the electron capture detector response to Aroclor 1254) Is 60 HONS 056281 about 0.2 ppm for individual foods and about 0.05 ppm for Total Diet composite. Fish from certain fresh water locations are the food most frequently and consistently found to contain PCB residues. The residues In fish are most often similar in gas chromatographic peak pattern to Aroclor 1254 but usually with higher concentrations of the late eluting (higher chlorination) chlorobiphenyl components. E. Chlorinated Dlbenzofurans (See Appendix for structural Information and definitions) In considering the potential hazard to humane of the commercial chemical mixtures of chlorinated biphenyls called PCBs, one must consider which if any identified trace contaminant In these complex mixtures might contribute significantly to the overall hazard potential of the mixture. Although there is evidence for the presence of chlorinated napthalenes and possibly chlorinated terphenyis In PCBs, the chlorinated dlbenzofuran (ClDBF) contaminants are regarded as a greeter potential danger for several reasons. In using a chick bloassay to monitor fraction ation of commercial PCB mlxtrues, it was found (Vos et si., 1970) that the fraction most toxic to chicks, and far more toxic than the other fractions, contained Cl-DBFs and chlorinated naphthalenes. What limited toxicological information there Is available on these halogenated naphthalenes, terphenyis and dlbenzofurans suggests that only the Cl-OBPs may be more toxic than chlorinated biphenyl* by orders of magnitude (Bauer et ai., 1961; Moore et al,,1976). 61 MGNS 056282 The extremely high toxicity of some of the chlorinateddibenzo-p-dioxins (NIEHS Conference 1973; Schwetz, et al., 1973) a class of organic chemicals very similar in structure to the Cl-DBFs, also suggests the latter are likely to be highly toxic. Knowledge of the chemistry and toxicity of various chlorinated dlbenzo-p-dloxln structures and the relationship between toxicity and structure, although recently developed. Is more complete than for Cl-DBFs. Therefore, findings in the chlorodloxln field provide indicators to assess the difficulties likely to be encountered in the study of Cl-DBFs. Chlorinated dibenzofuran contaminants have been reported and confirmed In PCB mixtures manufactured in Germany and France (Vos et al., 1970; Bowes, et al., 1973a) In Japan (Roach and Pooerantz, 1976a; Nagayama, et al.' 1975) and in the United States (Aroclors ) (Roach and Pomerantz, 1976b; Bowes, et al., 1975a). An additional claim that a cl-DBF contaminated an Aroclor was not sufficiently supported by the evidence pre sented (Curley et al., 1975). In the PCBs of U.S. manufacture, tho range of Cl-DBFs found was from dlchloro through hexachloro (see references cited above) and sufficient evidence was obtained to show that 2,3.7,8-tetrachloro and 2,3,6,7,8 -pentachlorodlbenzofuran are preson* In Aroclors (Bowes et ol., 1975b). That tho Cl-DBFs reported were actual contaminants and not artifacts resulting from the experimental procedures used, was considered in two instances (Roach and Pomerantz, 1976b; Nagayama et al., 1975). 62 MONS 056283 Proper quantitation of the Individual Cl-DBF contaminants la very difficult. It requires full structural identification of the Cl-DBP contaminant, availability of the contaminant an a highly purified reference standard for quantitation purposes and evaluation of the procedure used to concentrate and separate the Cl-DBFs, (e.g. from chloroblphenyla) to determine the capability of the procedure to recover the specific Cl-DBFs in the PCB mixture. Reported attempts to quantitate the Cl-DBFs In PCB suggest contamination levels in the low parts per million range for total chlorinated dlbenzofuran* (Bowes et al., 1975a; Nagayama et al., 1975). Many of the reported values probably can ba taken as minimal in the absence of recovery data. Although the source of the Cl-DBF contaminants in conmaercUl PCB mixtures has not been determined, several possibilities may be considered. The simplest explanation would be the likely presence of dibenzofuran (parent compound) In the technical grads biphenyl subjected to the chlorination process. From a consideration of the procedures used In the commercial synthesis of PCBi (Hubbard, 1964) various chlorinated biphenyls, If aubstltutad in the ortho and ortho-prime positions with hydroxy groups and/or chlorine atoms, might ring close to frm the furanold ring by dehydration or dehytk'ochlorln .cion. It Is important to recall that Cl-DBFs have bean reported aa contaminants also In other chemicals of conDerclal Importance. Samples of pentachlorophenol (Schwatz et al., 1974) aa well as 63 MOMS 05628*1 lower chlorinated phenols (Firestone et al., 1972) and hexachlorobenzene, (Villanueva et al., 1974) have boon found to contain a range oC Cl-DBFs. The ppm level of Cl-DBFs in certain pentachlorophenols examined by Schwetz and co-*workers (1974) was considerably higher than the highest level thus far reported in commercial PCB mixtures; the chlorine level In the reported dibenzofuran contaminants of pencachlorophenol ranged from hexa to octa. It is now known that "PCB residues" obtained from environ mental samples usually do not simply represent an easily Identified original commercial PCB mixture. For example, there Is evidence from the study of electron capture gas chromatograms that certain chlorinated biphenyl compounds are preferentially Lost or concentrated In passing through environmental media. This has led to recent studies of the toxicity, metabolism and physiological effects of specific, individual chlorinated biphenyl compounds, synthesized by routes Intended to yield a slngla com pound of known structure. In addition to permitting the development of structure-activity correlations, these studies presumably were intended to avoid complications In Interpreting results from the tasting of the commercial chlorinated biphenyl mixtures (Vos and Notenboom-Ram, 1',2), now known to contain toxic Cl-DBFs as contaminants. Interpretalon of results from the testing of individual chlorinated biphenyl compounds, however, may not be entirely straight forward. In the Ullmann coupling reaction of (>t* HONS 056285 2,4,5-trichloroUwUvbonr.'m*, It h* been i r^ot t #,1 <)i*t the l 1 iaU t v toxic 2, l,7,H ti*t rm-Morodibenzof nr an lit formed In 31 yield In addition to the expected 2,2'4,4',5.5'-hexachloroblphenyl product (Moron, et al., 1973). This hexechlorobiphenyl Is a significant constituent of PCB mixtures (Sissons and Eltl, 1971; Tas and Klelpool, 1972; Jensen and Sundstrom, 1974) and has been singled out for biological studies by several workers (Vos end Notenboom-Ram, 1972; Johnstone et al., 1974; Hansell and Ecoblchon, 1974). Careful purification and, If necessary, chemical analysis of symmetrical chloroblphenyls prepared by the Ullmann coupling reaction (for dlbenzofuran content) Is sug gested as a prerequisite to biological testing of the biphenyl. There are two other aspects of the chemical relationship betven chlorinated biphenyls and Cl*DBFs that naed to be mentioned. Both deal with possible environmental alteration. Evidence hee been obtained for photochemical conversion of certain chlorinated biphenyls to Cl-DBFs in very low yields (Crosby and Mollanen, 1973). Although the scope of this study was limited, formation of Cl-DBP was shown to occur under conditions of sunlight irradiation and In the laboratory using sunlight-simulating conditions. Two different ortho-chlorinated biphenyls (the 2,5-dlchloro and 2,2*,55'-tetrachloro) were claimed to produce approximately 0.2X steady-state yields of a monochloro DBF. The limited reports regarding the photochemical decomposition of Cl-DBFs (Crosby and Mollanen, 1973; Hutzinger et al., 1973) indicate relatively 65 MONS 056266 rapid destruction of the compounds may take place In the environment. Since reductive dechlorination Is a major route of photochemical alteration of Cl-DBFs, the possibility of dechlorination of highly chlorinated DBFs to more toxic DBFs of lower chlorine concent must be considered. The extent to which specific chlorinated biphenyls, found aa major constituents of commercial PCB mixtures, can be photochemlcally converted to Cl-DBFs can only be determined by further experimental studies. Analysis of the Yusho oil (the rice oil contaminated by PCB-contalning heat exchange fluid implicated in the Japanese "Yusho'* poisoning incident) by Nagayaraa et al., (1975) for Cl-DBF content, led to a value of 5 ppm Cl-DBFs. This value is about 300 times the C1-D8P level expected in the Yusho oil if one simply assumes contamination of the oil by Cl-DBFs to be proportional to the Cl-DBF level found in unused Kanechlor 400 (the Japanese PCB mixture claimed to have been used as the heat exchange fluid) and to the level of PCB In the Yusho oil. This led Kuratsune et al., (1975) to suggest that Cl-DBF levels increased in the heat exchange fluid through use. Colng a step further, these questions may be raised: Is Cl-DBF concentration increased, generally. In PCB-contalning heat exchange fluids through use? Does such Increase occur in PCB-contalnlng trans former fluids or electrical capacitors through uae? A critical review is needed of the published data upon which Kuratsune's suggestion Is based (Nagayama et al., 1975), particularly 66 MONS 056287 sine* the results from a recent analysis of a portion of Yoeho oil (Trotter, 1976) raises questions about the concentration of chlorinated biphenyls in the Yusho oil. Further laboratory studies suggested by such a review and analyses of selected, used PCB-containing industrial fluids might help to answer the above questions. Considering the Cl-DBFe as the toxlcologlcally most signifi cant class of chemical impurities In the PCB mixtures, the questions may be asked: what portion of the observed effects from animal and human exposure to PCB mixtures can be attributed to the Cl-DBFs? It la doubtful this question can be answered in any quantitatively explicit way. Thera are analytical problems in Identifying and measuring accurately the amounts of chlorinated dlbenzofurant In various PCB mixtures. Even though only a relatively few of the 135 possible chlorinated dlbenzofurans may be Identified in PCB mixtures, the toxicity of these compounds can be expected to very with both number and ring position of chlorine atoms In the Cl-DBF molecule. Toxicity data on specific, high purity Cl-DBF* Is very limited because the compounds themselves are not readily available. In addition to these, essentially chemical, difficulties, toxico logical judgements would be difficult to make. To begin to evaluate the toxicological "burden" to be placed on Cl-DBF contaminants in commercial PCB mixtures, a practical approach would be to use Analytical proceduras that can recover and quantitate specific Cl-DBF compounds known to be highly toxic. For example, 2,3,7,8tetrachlorodlbenzofuran, already reported by Bowes (1975b) as 6? HONS 056288 present in PCBs, can serve a a measure of potential hazard until further toxicity data for Cl-DBFs become available. To date, there have been no published, positive findings of Cl-DBFs In environmental samples or In foods. Analytical pro cedures to detect Cl-DBFa In such samples should be developed, tested and applied to appropriate samples. If chlorinated dlbenzo-p-dloxin residue analytical findings can be taken aa an Indicator, procedures suitable for parts per trillion detection of Cl-DRFs will be required and quantitation will be difficult. No clear evidence for the presence of chlorinated dlbenzo-pdioxlns in commercial PCB mixtures has been reported. A recent publication suggesting the presence of such contaminants In some PCBe and in a synthesized chlorinated biphenyl (Ax and Hansen, 1975) almost certainly misinterpreted the significance of the experimental finding related to the dioxins. Ill. Chemistry of Bromineted Biphenyls A. Comparison of Bromlnated Biphenyls with Chlorinated Biphenyls Unlike PCBs, the chemistry and stability of PBBs have not been well studied and documented In the literature. It la difficult to assess the stability and the extent of possible chemical conversion of PBBs in the environment. PBBe can be compared chemically to the PCBs. Since both bromine and chlorine are 68 HONS 056289 ^ halogens, PBB and PCB chemistry should be similar in some respects. Bromine, however, Is a better leaving group In chemical reactions than chlorine. The described laboratory experiments concerning PBB alkaline hydrolysis and photolysis show that bromine is more labile than chlorine under comparable reaction parameters. PBBs may, therefore, be less stable in the environment if PBBs were under the same physical reaction para meters in the environment as PCBs. The physical state and other physical reaction parameters of PBB and PCB residues In the environment can determine reactivity. Liquids and vapors often react more readily than sollda. Aroclors are liquids, though some Aroclors are extremely viscous at room temperature. Some PCBs, especially the lover chlorinated components, are volatile. FlreMaster BP-6, however, is a solid and has an extremely low vapor pressure. The production, distribution, and usage of PBBs has not been as wide spread as PCBs. PBBs,unlike PCBs, may not be physically located In a position for chemical reaction. FlreMaster BP-6 haa been extensively used, for example. In thermo plastics, such as typewriter and business machine housings (Kerst, 1974). FlreMaster BP-6 has little tendency to migrate from the thermoplastic into which It Is Incorporated. The major portion of the products into which FlreMaster BP-6 is incorporated Is assumed to be burled eventually In refuse dumps. PCBs, on the other hand, are often used in transformers and capacitors and can be exposed to high temperatures which can accelerate possible V- chemical reactions. PCB residues are found in many diverse locations 69 MONS 056290 throughout most of the world. Many of these locations may offer conducive settings for chemical and metabolic conversions. PCB residues in a body of water may react photolytically, whereas PBB residues burled in a refuse dump would aot be In a position to absorb light. B. Non-roetabollc Alteration of Bromlnated Biphenyls Oxidation and Hydrolysis The stability of PBBs to oxidation has not been studied and documented In the literature. FireMaster BP-6 Is unstable to alkaline hydrolysis. After refluxing FireMaster BP-6 with 21 KOH in ethanol* GLC chromatograms show erratic degradation of hexabromoblphenyl (DET*DOtDCT*DCH*l976), the major component of FireMaster B7-6. The possible rate of PBB hydrolysis In the environment under milder conditions Is not known. Photochemistry Recent analysis of FireMaster BP-6 by GCMS using an OV 101 column demonstrated 12 peaks whose mass spectrum corresponds to 2 pentas, 4 hexas, 4 heptas and 2 octas. However* about 50 percent of the material Is 1 hexa Isomer with the next most abundant component being the hepta Isomer at about 20-25 percent Nuclear magnetic resonance and chemical studies have Identified the principal component of Firemastor BP-6 as 2,2'4,4,5,5,-hexabromobiphenyi (Anderson et al., 1974; Sondstrom et al., 1976). 70 HONS 056291 The photolytlc rates of reactivity of FlreMaater BP-6 and Z^^^'S^'-hexachlorobiphenyi in methanol solutions irradiated with 3000 A light were compared (Ruzo and Zabik, 1975). The rate of reaction of FlreMaater BP-6 waa determined preaumably by the decrease in the GLC peak of hexabroaobipenyl. Hexabromoblphenyl waa found to be seven tinea aa reactive aa the corres ponding PCB. Hexabromobiphenyl waa found to undergo photolytlc reductive debromlnation in methanol yielding penta- and tetrabromoblphenyls and alao> IX methoxylated product. Dltnethoxy tetrabromoblphenyl waa identified aa a product by mass spectrometry. The highly efficient photoreactivity of PBB may be due to: 1. enhanced lnteraystem crossing to a triplet state due to vlbronic coupling with the bromines, 2. aterlc Interference due to the bromines, and 3. the relatively low carbon-bromine bond energy. The aromatic carbon-bromine bond energy la 71 kcal/mole vs the aromatic carbon-chlorine bond energy of 86 kcal/mole (Keret, 1974). As with PCBs (Ruzo, et al., 1974), presumably ortho halogens pre ferentially cleave in PBBs upon photoexcitation. The possible rates and extent of photolytlc reactivity of PBBe In the environment are not known. Tb photochemistry of PBBs has not been studied In the vapor or solid states. Since FlreMaater BP-6 has an extremely low vapor pressure, vapor state photochemistry in tha environment would be extremely limited. Tha extent of thin film PBB photochemistry in the environment may also be limited. PBB contamination and resulting 71 HONS 056292 photochemistry in river waters, except in the vicinity of PBB production facilities, may be limited. PBBs Incorporated Into thermoplastics, which are presumably burled eventually in a refuse dump, are not likely to absorb much light for photolytlc reaction. C. Bromlnated Biphenyls: Determination of Residues Polybromlnated biphenyl (PBB) residues are analyzed for in foods by methods quite similar to those used for organochlorlne pesticides and polychlorinated biphenyls (PCBs) (Food and Drug Administration, 1968-1975; Fehrlnger, 1975; Food and Drug Administration, 1976). Recoveries of PBB added to samples In vivo through these procedures is over 80 percent. A method to Improve extraction efficiency of PBB residue from dry, high fat animal feeds has been reported (Fehrlnger, 1975). The major difference from methods for organochlorlne pesticides Is In the gas chromatographic determination which is done at higher temperatures or on columns with low liquid phase loads because of the lover volatility of PBBs. Methyl silicones or Silar-lOC, the liquid phases most frequently used, separate the technical PBB, FlreMaster BP-6, into 3-9 peaks depending on the chromatographic conditions selected. PCBs and organochlorlne pesticides which would be recovered through extraction and cleanup procedures together with PBBs have earlier retention times under these conditions and are separated from the 72 HONS 056293 major Pbb cimeiituwiu, hwAabtumobiphenyi. The retention times or hexabromoblphenyl and decachlorobiphenyl are not greatly different on methyl silicone liquid phases but are widely separated on Silar-lOC. Elution of the Florlsll column used for cleanup of extracts with petroleum ether rather than a mixture of petroleum ether and ethyl ether separates PBBa from most organochlorine pesticides prior to gas chromatography and Improves cleanup for the PBB determination. PBB residues are detected and quantitated with the electron capture gas chromatography detector. Quantitation is baaed on comparison of the size of the hexabromoblphenyl peak In the residue to the size of the hexabromoblphenyl puak in a known weight of the reference material, FlreHaater BP-6. FlreMaeter BP-6 la a technical mixture containing 60-70 percent hexabromobiphenyle (Sundstroa and Hutlinger, 1976). The accuracy of residue quantitation la affected by any change in the composition of the residue from that of the reference technical PBB mixture. The identity of PBB residues can be confirmed by thin layer chroma tography, photochemical alteration, halogen-specific gas chromatographic detection, and by different retention times on Sllar-lOC and methyl silicon gas chromatographic columns (Food and Drug Administration, 1976; Fehringer, 1974; Fehrlnger, 1973; Emey, 1975). Mass spectrometry has also been used to confirm the identity of the PBB residue (Ayers, 1975). 73 MONS 056294 The limit of quantitation for PBB residues, including capa bility for residue identity confirmation, is about 0.05 ppm in fats and 0.01 ppm in non-fats (Food and Drug Administration, 1976). Formalized interlaboratory studies have not been reported for analytical methods for determination of PBB residues. D. Possibility of Bromlnated Dlbenzofurans in Commercial PBB Mixtures There has been no report, so far, of the finding of bromlnated dibenzofuran (Br-DBF) compounds in comercial PBB mixtures. Examination of FlreMaster BP-6 for possible contamination by Br-DBF is in progress In several laboratories. 74 MONS 056295 METABOLISM AND BIOCHEMICAL TOXICITY OK PCBs and PBBs I. Effects of PCBs on Biochemical. Functions One of the major biochemical effects of PCBs Is the Induction of microsomal enzymes In the liver. Rlsebrough et el., (1968) suggested that PCBs had the capability to Induce the activities of microsomal enzymes. Subsequently, Street et el., (1969) demonstreted the Induction of liver enzymes in rats by PCB's. Since then, a great number of articles have been published on this subject. The enzyme systems studied have Included mainly hydroxylases, N- and O-demethylases and nitroreductases, and to a lesser extent nonspecific carboxylesterase, bromosulphophthaleln-glutathlone conjugating enzyme, p-nitrophenol UDP-glucuronyl transferase and EPN-detoxificatlon systems. The Induction of microsomal enzymes by coaaerclal PCBs has been demonstrated perorally In rabbits (Vllleneuve et al., (1971), rats (Lltterat at si., 1972a) and primates Allan et al., 1974) and via lntraperltoneal injection (Bickers et al., 1972) and akin application In rats (Bickers et al., 1975). Values reported for the threshold of enzyme Induction by PCB'e vary betveen 0.5-25 ppm. (Lltterat et al., 1972b; Vllleneuve et al., 1971; Turner et el., 1974). The time-course of microsomal eyzyme Induction was studied by Lltterat et al., (1974) In rats. They found that significant levels of enzyme induction occurred after 7 daya of feeding PCBa 75 HOMS 056296 in the diet. A single Orel dose in rets resulted in maximum enzyme activities at 24 hours. Bickets at al., (1974); 1975) have shown that maximum induction occurred within 2-6 days in rats after cuta neous exposure to Aroclor 1254 or microscope lomerslon oils con taining 30-452 chlorine. In order to study the effect of chlorine content of PCB's on enzyme induction, different comerclal PCBs were Investigated. Lltterst et al., (1972b) studied PCBs with chlorine contents from 42 to 60X. In rats the maximum activity of demethylase vas observed with the 54X chlorine PCB's; maximum activity for nitroreductase was obtained with the 60X chlorine PCBs. Chen et al., (1973) reported that 54X chlorinated PCBS gave maximum response of demethylase in rats. Similar results were reported by lntraperltoneal injection of various PCBS in rats (Ecoblchon et al., 1974). Goldstein et al., (1975) and Iverson et al., (1975) studied the effects of Aroclor 1016 and Aroclor 1242 on enzyme induction in rats. Both PCBs have approximately 42X chlorine, but the content of the penta-, hexa-, and hepta-chlorlna Isomers of Aroclor 1016 is reduced to 10X of that in Aroclor 1242. Their results showed that Aroclor 1242 was a much more potent Inducer than Aroclor 1016 in female rata. In comparing Aroclor 1232, Aroclor 1248 and Aroclor 1260, Schmoldt et al., (1974) found that Aroclor 1260 was the most potent enzyma Inducer in rats. The synthetic and lsomerlcally pure PCBs have been investi gated for enzyma induction, Chan et al., 91973b) found that both 76 HONS 056297 2,3,4,5- and 3,5,3',5'-tetrachloroblphenyla induced enzyme systems In male rata, but no significant differences were found between the two Isomers. Johnstone et al., (1974); Ecoblchon et al., (1975); Ecobichon (1975) compared the effects of position of chlorine atoms on the ring. They found that enhanced Induction of mono-oxygenases was observed for PCEs having chlorine atoms substituted at the 4and 4'- positions irrespective of chlorination at other positions. Substitution et the 2-posltion was next in Importance followed by substitution at the 3-poaition. They concluded that the position of chlorination was as Important as the degree of chlorination. There are two types of enzyme Inducers being reported In the literature. One group to which phenoberbltal belongs resulted In increased cytochrome P-450 content, es well as Increased benzpyrene hydroxylase and ethylmorphine demethylase activities in the liver. The second group Includes polycyclic hydrocarbons. This group stimulates the formation of cytochrome P-440 and an increase In hydroxylatlon but not demethylatlon. Alvares et al., (1973) reported that rats treated with PCBs produced an Increase In cytochrome P-448, hydroxylase, as well as demethylase. There fore, it appears that PCBs display Induction behavior typical of both groups* Host of the enzyme Induction studies were evaluated In vitro. Several workers Indeed have observed enzyme induction effects in vivo by demonstrating shortened barbiturate sleeping times (Bickers et al*, 1972; Johnstone et al., 1974; Villeneuve et al., 1972): 77 HONS 056298 CommtirrUl Vi'.Wh r known to contain email amounts of dtbenzoiuran* (DBF). DfiFa were found to be 170 times as potent as PCBS In inducing enzymes. Alvales & Kappas (1975) reported the Induction of hydroxylase and demethylase In placenta, as well as In the fetus, when preg nant rats were treated with FCBs Valnlo (1974) reported that Clophen A50 enhanced hydroxylase 5-fold In the lung and 8-fold In the kidney mlcrosomes, whereas the microsomes from duodenal mucosa exhibited no enhancement. Benzpyrene hydroxylase activity In skin was also Increased by FCBs (Bickers et al., 1975). Vos and Koeman (1970) found that sevaral tissues of PCB-treated chickens were strongly fluorescent under ultra violet light. They suggest that PCBs could Indues chemical porphyria in chlckans. Later, they demonstrated that mitochondrial ALA Synthetase activity Increased 20-fold and fecal porphyrin levels were significantly Increased In Japanese quail treated with PCBs (Vos, 1971; Vos et al., (1972). They suggested that the porphyria caused by PCBs Is due to the Increase of ALA synthetase, followed by overproduc tion of porphyrlne. Similar results In rate were obtained by Goldstein et al. (1974), and Bruckner et al., (1974). Goldstein et al,, (1974) suggested that the induction of ALA synthetase la probably not the primary cause of PCB-lndueed porphyria because of the delayed onset of porphyria as compered with ALA synthetase induction. They suggest that the ALA synthetase In the liver may ba ralatad to tha increase of cytochrome P-450 content. This 78 HONS 056299 idea was supported by Grote et al.( (1975). Aroclor 1242 was found Co be a more potent Inducer of liver porphyria than Aroclor 1016 (Goldstein et al., 1975). Both Aroclors have similar chlorine content, but Aroclor 1242 contains 9X homologs with five or more chlorines while Aroclor 1016 contains only IX of such homologs. Sinclair end Granlck (1974) showed that cultured liver cells respond rapidly to PCBs in accumulation of uroporphyrin. Simon et al., (1974) suggested that porphyria may be due to the destruc tion of the phospholipid structure of cell membrsne, resulting in Increased permeability of the substrate. Hlrayama et al., (1974) observed hypo-blllrublnaemla in Yusho patients and suggested that the induction of bilirubin UDP-glucuronyltransferase may be respon sible for the low plasma bilirubin content. However, recently Bastomsky et al., (1975) showed that the lowering of serum bilirubin concentration by PCBs is due to the reduced binding of bilirubin to plasma protein rther than to the enzyme induction. Wit (1972) has shovm that PBBs which are similar to PCBs are potent heptoporphy-rlnogenic chemicals. The chemical and structural similarity of DDT to PCBs led Jefferies at al., (1972) to study the effects of PCBs on thyroid functions. They found an Increase in thyroid weight in PCBtreated black-backed gulls as compared to controls. Hurst et al.. (1974) found no clear-cut effect of PCBs on thyroid gland size in quails. Byrne et al., (1975) reported that PCBs increased 79 HONS 056300 thyroxine levels in the blood and also promoted peripheral degrada tion of thyroxine in female minks. Bastomsky (1974) found a 4 to 5-fold Increase of biliary excretion of thyroxine In rats treated with FCBs. PCBs also elevated Iodine uptake by the thyroid and reduced serum PBI concentration. Recently Bastomsky and Wise, (1975) showed similar results in rats due to either intraperitoneal injection or skin application of microscope Immersion oil. Walker (1975) found that inclusion of Aroclor 1254 in fish food resulted in an Increase in fish thyroid activity. The effects of PCBs on ATPase and oxidative phosphorylation have been examined by a number of Investigators. Na4TCf-ATPaee and Mg*+ ATPase from several fish tissues were found to be Inhibited by PCBs. (Yap et el., 1971; Cutkomp et al., 1972; Desalah et al., 1972; Koch et al., (1972). Further study shoved that in vitro data were not in agreement with the in vivo data (Yap at al., (197i). The inhibition of ATPaae in fishes by PCBs was also reported by Kinter et al,, (1972) and by Davis et al., (1972). Sivalingan et al., (1973) investigated the effects of PCB on the oxidative phosphorylation of rat liver mitochondria. They shoved that the mode of inhibition appears to be different with PCBs of different chlorine content. PCBs with low chlorine content inhibited energy and electron transfer but PCBs with high chlorine content not only inhibited energy and electron transfer but also had an uncoupling effect. LaRocca et al., 80 MONS 056301 (1975) showed chat Che administration of commercial PCBs, as well as purified Isomers to rata Inhibited the total ATPase activity in liver, kidney and brain tissues. Pardlnl (1971) has demonstrated that a marked inhibition of respira tory enzyme systems occurred when heavy beef heart mitochondria were exposed in vitro to numerous PCBs. Chesney and Allen (1974) showed that the addition of PCBs to rat liver mitochondria in vitro caused an inhibition of oxidative phosphorylation and respira tion. Feeding rats with PCBs at the 1000 ppm level Increased the oxidative phosphorylation In liver mitochondria, however no affect was seen at the 100 ppm level. Sharp et al., (1974) reported that the inhibition of beef brain and rabbit kidney Na+lC^-ATPases can be reversed or prevented by phosphetldylserine or phosphetidyllnoeltol, but not by phosphatldychlollne and phosphatidylethanolamine, They suggested that acidic phospholipid# are required to stabilize the enzyme and thereby overcome the effect of PCBs. Rieebrough et al., (1968) suggested that PCBs had the capacity to enhance steroid-hydroxylating enzyme activity, thus affecting estradiol metabolism. In addition, certain low chlorine containing PCBs have estrogenic activity which is reflected by en Increase in glycogen content of the uterus of immature rats (Bltman end Cecil, 1970). Similar results were obtained by Fumiko (1972) in rate (Bltman end Cecil, 1970). Similar result# were obtained by 81 HONS 056302 Fumiko (1972) In rats and by Orberg, and Kihlstrom (197 ) In nice. Orberg et al., (1972) found that the estrous cycle of the mouse Increased In length after a single injection of PCBa. Linear and Peakall, (1973) reported an increased rate of metabolism of estra diol in vitro in the liver of American kestrels after the feeding of PCBs. Similar results were also reported In white leghorn cockerels and pullets (Novlckl and Norman 1972). Platonow and Funnell (1971) demonstrated that feeding PCB* to cockerels resulted In a decrease in testicular and comb growth. Decreased urinary excretions of estrogen and dehydroeplandroaterone were also observed in cockerels when PCBs were administered in high doses (Platonow et al., 1972). Ecoblchon and Mackenzie, (1974) studied the uterotropic activity of commercial and isomerlcally pure PCBs in rats. They found significant changes with a number of PCd mixtures but experiments with the isomerlcally pure PCBs were inconclusive. Changes were observed with 2-chloro and 2,2chloroblphenyls. Nelson (1974) reported that PCBs are effective inhibitors of the binding of ^H-estradlol to the rat uterus cytosol fraction in vitro. Recent data showed that PCB feeding caused neither significant chromosomal damage nor arreat in the rate of spermatogenesis in male rats (Dikshith et al., (1975). Flick et al., (196S) noted enlarged adrenals and small spleens in PCB-treated white leghorn cockerels. Wassermann and Wasaermann (1972) and Wessonoann et al., (1973) reported that rats receiving 200 ppra PCBs showed an increase in plasma corticosterone levels. 82 HONS 056303 These changes were concimitant with morphological changes in the zona fasciculate of the adrenal gland which indicated Increased activity. Sanders, et al.f (1974) have shown similar Increases in serum corticosterone in mice fed 60 to 1000 ppm PCBs. It le known that DDT decreases the utilization of dietary carotene and liver storage of vitamin A in rats and cattle (Phillips, 1963; Phillips, and Hidlroglou, 1965). Villeneuve et al., 1971) reported that Aroclor 1254 but not Aroclor 1221 reduced the liver concentration of vitamin A in pregnant rabbits. Decreased liver vitamin A concentrations were found in male and female rats and also Japanese quail after PCB feeding (Cecil et al., 1973). How ever, the laying female quail did not appear to be affected. The effects of PCBs on vitamin E have been studied by Combs at al., (1975). Their results indicated that dietary PCBs Increased the incidence of exudative diathesis in chicks and that this Increase can be overcome by increasing dietary vitamin E or selenium. The vitamin D mediated calcium metabolism vas found to be altered by the oral administration of PCBs to chickens (Wong et al., 1974). II, The Comparative Biochemical Toxicity of Aroclor 1254 and Plremaater BP-6 PCB'a have been reported to affect a large number of bio chemical systems. To the extent thet It has been reported, PBBs 83 HONS 056304 affect moat of these same systems in a similar fashion. However, quantitative and possibly qualitative differences do not appear to exist in the response of some biological parameters. These include thyroid hyperplasia (Norris et al.* 1973) and microsomal enzyme induction (Farber and Baker, 1970). A preliminary study was conducted to compare the relative effect of Aroclor 1254 (Aro) with Firemaster BP-6 (FM) (Garthoff et al., 1975). Tills was set up as a broad survey to give an estimate of the relative biological activity of these chemicals In may different systems and pin point areas where further more definitive work should be done. Table 1 summarizes the time sequence of the various parameters studied. Adult male rats were fed either 0, 5, 50, or 500 ppm Aroclor 1254 or Firemaster BP-6, mixed in the diet, for 2, 3, or 5 weeks. Due to the large number of parameters examined and the time restraints placed on the study, the experimental design did not permit a totally unambiguous comparison between ell of the effects caused by Aro and FM. Some apparent differences In biochemical responses determined at sacrifice may have been partly due to the fact that, for the three-week study, rats were exposed to FM for 48 hours longer than rats were exposed to Aro. Although it is felt that under the experimental conditions, this effect should be small, critical comparison of Aro and FM toxicity should be 84 MONS 056905 considered tentative until smaller experiments are conducted which can eliminate this difference in exposure tine. The following evaluations were made based on the assumption that this two day difference had no effect. Some parameters showed no consistent or dose related change with either chemical at any level. These Included relative kidney weight and testes weight, hematology, SGPT, SCOT, plasma BUN, plasma corticosterone* the rate of liver protein synthesis per g tissue, liver dry weight, lyophlliaable kidney, liver, or testes, the Incidence of chromosome abnormalities end the number of cells In mitosis from bone marrow of spermatogenial cells. Growth snd adipose tissue weight were equally depressed by Aro and FM when fed over e three week period at 500 ppm* The growth effect appeared to be due primarily to decreased food efficiency. Three parameters were more sensitive to Aro than to Fm. These included the decreased liver protein end liver RHA per g tissue and the decreased plasma glucose. On the other hand, six parameters were more sensitive to FM. These included liver growth, Increased liver lipid (total lipid, cholesterol* phospholipid, and neutral lipid), elevated plasma cholesterol, Increased microsomal ensyme activities, impaired mitochondrial function, and decreased liver RNA synthesis. 85 HONS 056306 Disruptions of the redox and energy states of the cell were produced by dietary exposure to Aro or FM. These effects were Indicated by studies wit Isolated mitochondria and frozen damped liver. In the latter case the concentrations of adenine nucleotides, pyridine nucleotides, and glycolytic Intermediates were determined. On a molar basis, hexabroraoblphenyl was reported to be about five tinea more potent than Arochlor 1254 in causing some Increased microsomal entyme activities (Father and Baker, 1974). A three-fold difference was observed between Flremaster BP-6 and Arochlor 1254 (Table 3). A pathology team autopaied rats after both three and five weeks exposure to Aro and FM (K&sta et al., 1975). Pathological analysis of the liver of rats fed Aro or FM for three weeks shoved no abnormalities on gross observation. No histopathological lesions were seen in controls, Aro groups, or the 5 and 50 ppm FM groups, after three weeks. In the 500 ppm FM group, minimal vacuolation and focal hepatitis were each seen in one rat. After five weeks exposure, gross observation of the liver indicated an increasing incidence of enlargement, friability, and prominent lobular pattern with increasing dose of both chemicals. Micro scopically the 5 ppb Aro group appeared normal while 2/7 rats fed 5 ppm FM had minimal cytoplasmic degeneration. All rats had this lesion at 50 and 500 ppm Aro or FM, with Increasing cellular lipid. This losion was recent, centrllobular and coalescing with no change in fibrous tissue or glycogen. The hlstopathology was qualitatively similar but slightly more severe in rats fed FM 86 MONS 056307 than in those fad Aro and la consistent with biochemical findings. The biochemical findings were all made after three weeks exposure at which time hlstopathologlcal changes were not evident. Even at five weeks the hletopathology was not severe enough to alter most of the clinical chemistry in the rat. In most parameters evaluated, FM either caused sn effect et a lower doae level or ceueed an effect In a shorter time or caused a greater effect at the same dose level, expressed as ppm In the diet, as Aro. When expressed on a molar basis FM becomes evsn more effective than PCB. Exceptions to this observation were noted at the 500 ppm dose level where differential effects of Aro and Fm were often eliminated. A sumaary of the major effects seen ere found In Tables 2, 3, and 4. In all but one caae the maximum responee of a parameter occurred at the highest doss level. Microsomal ensyms activities were, in general, greattr at the 50 ppm level and the decline at 500 ppm relative to the 50 ppm level was mors marked with FM. Ill. Metabolism of PCB and PBB Mixture Generally, the tlesues from animals and man containing PCB from environmental exposure have GLC patterns resembling those of PCB mixtures with more than 50% chlorination. This la In marked contrast to the major manufactured products that generally contain 42% or lees chlorine. While one cannot rule out the poaalbllty that differential usee favored Introduction of more highly 87 HONS 056308 TABLE 1 PARAMETERS EVALUATED IN STUDY EXPOSURE TIME (WEEKS) PARAMETERS TWO THREE FIVE Body weight Liver weight XXx X Xx Other tissue weights X Mitochondrial respiration X Liver composition X Liver protein synthesis X Liver RNA synthesis X Liver Microsomal enzyme activities X Liver Intermediary metabolites X Serum cholesterol X Serum enzymes XX Plasma corticosterone X Plasma protein X BUN X Cross pathology XX Kistopathology Cytogenetic analysis XX X 86 HONS 056309 ( f TABLE 2 RELATIVE BIOCHEMICAL RESPONSE TO Aro AND FM AFTER TWO WEEKS DIETARY EXPOSURE PARAMETERS Liver velehtr Body weight Mitochondrial respiration Aro TJCT.J- ppa FM 500 50 500 500 Maximum Effect Z Control Aro FM 170 183 152 192 OT 9 5 0 SNOW - lowest effective level that shoved a dose-response relationship. TABLE 3 RELATIVE BIOCHEMICAL RESPONSE TO Aro AND FM AFTER THREE WEEKS DIETARY EXPOSURE LElA PP Aro FM Maxiautn Effect Z Control Aro FM PARAMETERS Body weight gain 50 500 52 52 Liver weight 50 Body weight 50 203 214 Liver dry wt. NE3 Liver lipid 50 Liver cholesterol 500 Liver protein 500 50 5 50 NE NE3 138 107 156 226 232 84 NE Liver RKA Liver DNA Liver RNA snythesls 5 5 500 50 5 5 79 84 58 60 78 86 HONS 0 5 6 3 1 1 vO c ( TABLE 3 (Cont'd) LEL3 ppe Aro FH PARAMETERS para-Hitrobenxoate reductase activity 50 Plasaa glucose 50 5 NE3 Plasaa cholesterol 500 5 Maxiuua Effect X Control Aro FM 2882 5202 79 NE3 178 256 ^Lowest effective level that shovs a dose response relationship. ^Kaxlaua effect occurred at 50 ppa dose, ^ - Ho effect HONS 0 5 6 3 1 2 TABLE A RESPONSE OF RELATIVE LIVER VEIGH'l TO Aro AND FM AFTER FIVE WEEKS DIETARY EXPOSURE* PARAMETER Liver weight body weight LEL PP* Aro FM 500 50 Maximum Effect Z Control Aro FM 171 216 * Legend - see Table 3. HONS 0 5 6 3 1 3 'fOs> chlorinated materials in the environment, the observation has led to the general belief that the less chlorinated components are more readily metabolized. The environmental observations have been confirmed in a large number of experimental feeding studies in a nuofcer of manuallan and avian apecles. The abaence or diminished concentretion of the early eluting peeks have been reported In rate (Grant et el., 1971a, Curley et el., 1971), rabbit (Grant at al. 1971b), cow (Fries et el., 1972a), quail (Bailey and Bunyan, 1972), and htna (Fries et si., 1973b). The peaks not prasent in tissues and products generally are the early eluting peaks that correspond to the PCBs with lower degrees of chlorination. The obaervatlona are conaletent with the belief that the rate of metabolic attack on PCBs decreases with Increasing chlorination. Studies of the single PCB homologs of various degrees of chlorination have ahovn that thoee with five or leas chlorine atoms ere more radlly metabolized and excreted than the PCBs with higher chlorination (Berlin at al., 1975; Hutzlnger et al, 1972; Melvas and Brandt, 1973). The position of chlorine substitution also affects t..e retention and elimina tion of single homologg (de Freitas and Norstrom, 1974). Metabolism of Individual PCBs will be discussed more thoroughly In another section. 93 HONS 05631** Among the various food species there appears to be one exception to the goneral rule that the leas chlorinated PCBs are hydroxyl.tied. The PCB residues In fish closely resemble the PCB to which the fish was exposed. There was little change in the^ relative concentrations of the various homologs (Stalling and Mayer, 1972). Consistent with this observation It has been found that mono**, di~, and tetrachlorobiphenyls are not metabolized by trout (Hutzinger et al, 1972) Among the food producing animals (cattle, chickens) concentra tions of PCB residues in milk and eggs reach a "steady state" after approximately 8 to 12 weeks of continuous Intake (Fries et al. 1973a; Fries et al., 1973b). The ratio of residue levels in food products to levels In the animal's diet are greatest with PCBs of 54% chlorination. The residue levels in milk are approximately 4 times the residue level In the cow's diet. In hens, residue levels in eggs are approximately equal to the diet while residue level in the body tissue (fat basis) is approximately 6 times the dietary level. The ratio of residue level in product and tissues to Intake is lower with PCBs of greater than 54% chlorina tion. Since it is unlikely that there is greater metabolism of the most highly chlorinated PCBs, the results suggest that the absorption for the Cl tract is lover for the more highly chlorina ted PCBs (Smith et al., 1976). PCBs of four or fewer chlorines rarely occur in animal products when the animals are fed environmen tally realistic levels. 94 HONS 056315 Much less 10 known about the metabolism of PBB than PCBe. How ever, the observations that have bean reported tend to be consistent with the report* on PCB metabolism. While the PBB of moat Immediate concern for human health (BP-6, Michigan Chemical Co.) la a multicomponent mixture like the PC8s, It la much simpler than the ordinary commercial pCB mixtures. The major component la a single hexabromoblphenyl accounting for over 60Z of the total material. The only other component occurring in significant amounts ie a single heptabromoblphenyl. In general, the measures of retention in edible tlseuea and excretion in edible products are similar for the haxabromoblphenyl and the hexachloroblphenyls (Fries at al., 1976; Fries and Marrow, 1975). Heptabromoblphenyl retention and excretion is only 1/10 of that of the hexabromoblphenyl. In cattle that have been heavily contaminated with BP-6, the heptabromoblphenyl was barely detec table (silk or tiaauea) 6 to 8 months after the time of Ingestion even though the hexabromoblphenyl level was over 1000 ppm (Pries et si., 1975). Since the heptabromoblphenyl la not more likely to be metabolised by the animal than the hexabromoblphenyl, It la reasonable to assume that its absorption la lower or its elimination through bile la greater than the hexabromoblphenyl component. Over 60X of octabromoblphenyl la excreted in the face* as the parent compound whan it la fad to rats (Norris et al., 1974). This observation supports the conclusion that there Is lass absorption with increasing halogenation. 95 HONS 056316 IV. The Metabolism of Individual PCBs Quantitative studies of the distribution of PCBs Indicate that most, if not all, PCBs which contain six or fever chlorine atoms are efficiently abosrbed from the gut of higher animals (Albro and Flshbein, 1972; Matthews and Anderson, 1975a; Van Miller et al., 1975). Furthermore, only about one-tenth of the absorbed dose Is excreted prior to metabolism to more polar compounds (Matthews and Anderson, 1975a; Van Miller et al., 1975; Balin et al., 1975). A number of the less chlorinated PCBs (l.e., mono-, dl-, trl-, and tetrachloroblphenyls) are readily metabolised and excreted by birds and mammals. There is little evidence that fish can metabo lize any PCB. It has been adequately demonstrated that a number of maamutlian species and the pigeon can readily degrade and excrete monochloroblphenyls, primarily 4-chlorobiphenyl (Matthews and Anderson, 1975a; Block and Cornish, 1959; Hutzinger et al., 1972; Safe et al., 197Sa,b,c). However, the rate of metabolism drops sharply as the number of chlorines per molecule Increases. For example, the rate of metabolism and sxcretlon of several PCB Isomers has been shown to decrease as the number of chlorines increases in the following order: 4-chlorobiphenyl 4,4'-dichlorobiphenyl 4,4'-dichlorobiphenyl 2,5,2',5'-tetrachloroblphenyl 2,4,5, 2',5'-pentachlorobiphunyl (Matthews and Anderson, 1975a; Van Miller et al., 1975; Matthews and Anderson, 1975b). The rate 96 HONS 05631? of metabolism and excretion of 4,4'-dichlorobiphenyl la approxi mately one-half that of 4-chloroblpheayl, la each eeee the PcRe are metabolized primarily by hydroxylet ion and conjugation with glucluronlc acid (Matthawa and Andsrson, 1975a; Balia at al., 1975). The importance of tha route of excretion, i.e., urine or feces, la determined primarily by tha degraa of chlorination. Approximately 60, 30, and 7 percent of a alngls doe# of a mono-, a di-, and a pentachloroblphenyl, reapactlvely, vara excreted In the urine of rata. The remainder of the doae was excreted In the feces (Matthews and Anderson, 1975a). Once the number of chlorine atoms on the biphenyl molecule reaches four, the position of the chlorine atoms becomas a mors Important factor in determining the rate of metabolism end excre tion (Matthews and Anderson, 1975a; De Fraltas and Norstrom, 1974). As was observed with the chlorinated benzene* (Jondorf et al., 1955; Jondorf et al., 1958) those PCBa which have two adjacent unaubatltuted carbon atoms ere metabolised end excreted more rapidly than PCBs having the seme degree of chlorination, but which do not have two adjacent unaubatltuted carbon atoms (Matthawa and Anderaon, 1975a). It la clao likely that, as with the chlorinated benzenes, tha absence of two adjacent unaubatltuted carbon atoms has a greater effect ea the degree of chlorination Increases. For exampla, the initial half-life of 3,5,5' ,5'-tatrachlorobiphenyl la only about two-fold graater 97 HONS 056318 in the rat (Matthews and Tuey, 1976) than that of 2,5,2* 5'tetrachlorobiphenyl In the same species (Van Miller et al., 1975). On the other hand, the Initial half-life of 2,4,5,2*,4*,5'-hexa~ chlorobiphenyl appears to be infinite, whereas the initial half life of 2,4,5,2*,5'-pentachlorobiphenyl Is only about two days (Matthews and Anderson, 1975a). Tills particular hexachloroblphenyl has been shown to be In the highest concentration of any PCB found in the adipose tissue of the Swedish population (Jensen et al., 1974). The Importance of two adjacent unsubstituted carbon atoms to the metabolism of PCBs may be derived from their effect on the formation of arene oxide intermediates during the metabolism of thesa compounds by the hepatic mixed-function oxidases. Evidence for the formation of arene oxide Intermediates In PCB metabolism was first provided by Gardner et al., (Gardner et al., 1973) in their study of the metabolism of 2,5,2*,5*-tetrachlorobiphenyl and in a later study of 4-chloroblphenyl metabolism by Safe et al., (1975c) which was specifically designed to provide evidence of an arene oxide Intermediate. Metabolism of PCBs which do not have two adjacent unsubstltutad carbon atoms by hepatic mixed-function oxidases, via direct hydroxylatlon of the biphenyl molecule and/or the formation of an arene oxide between a chlorinated and an unsubstituted carbon atom could occur, but each of these mechanisms would be expected to be slower than arene oxide forma tion between two unsubstltuted carbon atoms (Daly et al., 1972; Jerlna and Daly, 1974). The latter mechanism would be expected 98 MONS 056319 to result in dechlorination and/or shifting Of a chlorine atom (Daly at al., 1972). The metabolism of selected hexachloroblphenyls has been reported (Jensen and Sundstrom, 1974; Hutzinger et al., 1974; Haas, unpublished), and in two incidents (Hutzinger et al., 1974; KcKinney et al., unpublished) dechlorination and/or chlorine shifting vas observed, but in all casea the rate of metabolism was quite elov. Thus, those PCBa which are more readily metabolized and excreted may alao more readily form arena oxides. Certain arena oxides have been implicated as potential carclnogene (Daly et al., 1972; Jerlne and Daly, 1974) ae have certain PCB formulations (Ito et el., 1973; Kimbrough and Linder, 1973; Kimbrough et al., 1975), but there la no published evidence which indicates that the leas chlorinated PCB formulations have been ao rigorously testad. 99 056320 HONS ANIMAL 7'OXICOLOGY Introduction A number of reviews on different aspects of the toxicology of the PCBs have been written recently and no effort has been made In this report to give a complete outline of the animal toxicology (Kimbrough, 1974; Peakall and Risebrough, 1975; Nelson et al., 1972; Flshbeln, 1974). The material summarised In this report consists predominantly of more recently published information on toxic effects of PCBs in mammals and experimental studies in birds. Since Nelson et al. (1972) discussed the toxic effects produced in fish by PCBs extensively, the reader is referred to this and the other reviews cited above for information on toxicity of PCBs in this species. The emphasis of the present review was placed on chronic toxic effects wherever possible. Unfortunately, very little information was available for halogenated dlbenzofurans and the bromlnatcd biphenyl mixtures. Acute Toxicity Polychlorinated Biphenyls (PCB) The acute oral Lf)^Q of the PCBs in rats, rabbits and mice ranges from 1 to 10 g/kR body weight (Tables There is some indication that young animals may be more sensitive than adults, 100 HONS 05632i and female* nor* sens1t1ve than male* (Table 5). According to the claaelflcatlon used by the American Industrial Hygiene Aaaoclatlon (Hodge A Sterner, 1949), the acute toxicity of the PCBs can be claeeed ae slightly toxic (0.5 to 5 g/kg body weight) to practically non-toxic (5-15 g/kg body weight). The symptoms assocatlated with administration of a toxic dose of PCI (Aroclor 1242) In rats consist of diarrhea, diminished exploratory behavior, decreased response to pain stimuli, chromodacryarrhea, adlpala, oliguria, anorexia, erythema of limbs, followed by ataxia, coma and death. Death occurred up to 14 days following administration of the toxic dose. 24 hours after administration of a single toxic dose of Aroclor 1242, gross pathology Indicated all organs appeared normal except the liver and kidneys. No inf lamat ion was observed In the abdominal and Intestinal mucosa. Hlstopathologlcal findings shoved large discrete sudanophlllc vacuoles In the hepatocytas. Widely scat tered foci of tubular epithelial cells were present in the kidneys (Bruckner, et al., 1973). However, Kimbrough (1974) reported ulceration of the gastric and duodenal mucosa In rats after single oral doses of 3,000 mg/kg Aroclor 1254 or 1260. Only one metabolite of PCI Is known to have been tested for the acute LD^q* The 5-OH metabolite of 2,4,3'4'-tetrachloroblphenyl was known to be more toxic than the parent compound (0.43 g/kgBW/2.15 g/kgBW (Table 5). Dermal toxicity of PCB la summarized In Table 7. 101 HONS 056322 Polvbroalnitad Biphenyls (PBBa) Polybromlnated biphenyls shov * low order of scute toxicity to the ret (Tabla 8). In the ceee of chicks, e J7X mortality wee reported when P8B was administered In the diet at a level of 100 ppm, for IS days. Seven out of eight guinea pigs fed SO ppm dietary PBB died (Voe end VanCerderen, 197}). Effecte noted included cortical atrophy of the thymus, adrenal enlarge ment, marked depletion of the follicles and perlerterlolar lymphocytes sheath of the spleen. Chlorinated Dlbenzofurane See subacute toxicity. Chlorinated Hapthalenea Sea and of chapter. 102 HONS 056323 -r TABLF 5 Co&pouod Tested Species Route g/kg body vcLOen Aroclor 12S4 " 1260 " 125* " 1260 - 125* * 1221 " 1262 " 12*0 " 1254 M 125* " 125* ** 125* jCaneclor-400 Kaneclo r-400 taneclor-400 Kaneclor-400 Kaneclor-300 iCanecior- 300 BP-200 (biyheayla of dicloride aed belov 2.4'-dichlorobiphenyl trlchioroMpheayl biphenyl of trichloride and belov 2,*.3* ,4*-tetracr.lorobiphenyl 5-OS derivative of 2,*.3'*'- teirachlorobipheoyl 23*3**'-pentachloroblpheny1 Rat(Adult-Sherman Strain) Rat(Adult-Sherman Strain) Rat(veanling " ") Rat(weanling " M) Rat(fesale " "') Rac(fecale " ") 8at(female H ") Rat Rat(Vistar-30-day-old M-F) Rat(Vlstar-60-day-old M-F) Rat(Vlstar-120-day-old M) Rat(Wistac-120-day-old F) Rat(Uistar Strain - Male) Rat(Ulstar Strain - Female) Mice(CFI Strain - Male) Mice(CFI Strain - Female) Rat(Wiatar Strain - Male) Rat(Vistar Strain - Female) Mice(dd-atralr.-female) oral oral oral oral Intravenous oral oral oral oral oral oral orol oral oral orol oral oral oral oral Klce(dd-straln-female) Mice(dd-scrain-female) Mice(dd-strain-female) oral oral oral Mlce(CFI strain) Mice(CFI strain) In craper1tonea1 " Kice(CFl atrain) ** *-10 *-10 1.295 1.315 0.35S 4.00 11.3 4.25 1.3 1.4 2.0 2.5 1.30 (ml/Kg) 1.14 (ml/Kg) 1.875 (el/Kg) 1.57 (ml/Kg 1.15 1.05 6.36 Linder, et al. (1974) Panel on Hazardous Trace Substances (1972) Bruckner, et al. (1973) Grant * Phillips (1974) Ikeda, et al. (1970) Kitamura (1972) 7.86 3.06-4.25 9.27 " 2.15 0.43 Yamacoco & Toshiouta (1973) 0.65 o HONS 0 5 6 3 2 4 TABLE 6 Oral LDc.n (Rat) (Panel on Hazardous Substances, 1972) Compound Tested Aroclor 1221 Aroclor 1232 Aroclor 1262 Aroclor 1268 Aroclor 1260 Aroclor 1262 Aroclor 1268 (undiluted) (undiluted) (undiluted) (undiluted) (50Z soln in corn oil) (50X soln In corn oil) (50Z soln in corn oil) g/Kg body vt. 3.98 4.67 8.65 11.0 10.0 11.3 10.9 MONS 0 5 6 3 2 5 TABLE 7 Skin HiDsn Rabbita (Panel on Hazardous Substances. 1972> Coapound tested Aroclor 1221 Aroclor 1232 Aroclor 1242 Aroclor 1248 Aroclor 1260 Aroclor 1262 Aroclor 1268 (undiluted) (undiluted) (undiluted) (undiluted) (502 solo in corn oil) (50Z soln In corn oil) (33.3Z sola in corn oil) &/Kg body vt. >2.000 >1.26 > .794 > .794 >1.26 >1.26 >2.5 <3.169 <2.0 <1.269 <1.269 <2.0 <3.16 Compound Firemaster BP6 Octabromobiphenyl Octabromobiphenyl Hexabromobipheny1 Occabrooobiphenyl Species Rat Rac Bobvhite Quail Rabbit Rabbit TABLE 8 LD^n Polybrominated Biphenyls Route Oral Oral Oral Dermal Dermal LD50 21.5 g/Kg More than 2 g/Kg More than 12.5 g/Kg More than 5 g/Kg More than 10 g/Kg Reference Hill Top Research, Inc. <1970) Michigan Chemical. Company Aftosmis, et a^., 1972 " " MOMS 0 5 6 3 2 ? oo SUB-ACUTE AND REPRODUCTIVE EFFECTS OF PCB'S, PBB'S AND CHLORINATED DIBENZOFURANS A. Mammalian Sub-acute Effects of PCBs In contrast to the acute toxicity (oral and dermal) of the PCBs which is of relatively low order when the aubetancea are administered aa a single dose* aspects of sub-acute toxicity of both the PCBs and individual chlorinated biphenyls appear to be of far greater concern exhibiting species sensitivity and cumulative toxic effects following continuous exposure et low Levels. When groups of mice were given toxic rlcs bran oil containing 1,600 ppm PCBs or 5000 ppm of PCB mixture with 48X chlorine, the toxicity response wee similar in the two groups (Niehlzuml 1970). In e group of rate fed 2000 ppm of Phenochlor DP-6, deaths occurred between the 12th end 26th days with enlarged livers, atrophy of the spleens end e progressively Induced hepatic porphyria was noted (Voe end Koemtn, 1970). Administration of 1000 ppm of Arochlor 1254 in the diet of rets resultad in deaths between the 28th day end 55rd day (100X mortality) of feeding (Tucker end Crabtree 1970), 50X mortality at the same dosage in an 8-month study wee reported by Kimbrough et al. (1972). 107 MONS 056328 Two groups of 52 male Sherman strain weanling rata were fed 100 ppm Aroclor 1242 (6.6-3.89 mg Aroclor/kg body weight) and 100 ppm Aroclor 1016 (6.9-3.5 mg Aroclor/kg body weight) respectively for 6 months and 54 rats were fed ground chow. Groups of A rats were sacrificed at intervals. Microscopic examination of the liver showed evidence of lipid accumulation, enlarged liver cells, inclusions In a number of livers and hemorrhage and necrosis in one animal per experimental group each. No difference In effect between the two compounds was noted (Burse et al., 197A). Rats given 100 mg Aroclor 1242/kg body weight by stomach tube every other day for 3 weeks showed histopathologlcal changes only in the liver and kidneys aa foci of sudanophlllc vacuolation but no overt sign of toxicity (Bruckner et al., 1973). Rats (Sprague-Dawley) given 100 ppm of 2,5,2', 5'-tetrachloroblphenyl in the diet for 3 weeks showed less liver hypertrophy and fewer biochemical changes than rats given a similar dose of Aroclor 1248 (Allen et al., 1975). Administration of 100 mg/kg body weight of Aroclora 1242, or 1254 orally to rabbits once a week for 14 weeka resulted in enlarged livers with apparent destruction of the rough endoplasmic reticulum and strophy of the uteri, while Aroclor 1221 did not cause this effect (Roller and Zlnkl, 1973). 108 HONS 056^29 The umieual sensitivity of mink to PCBs wee reported by Aulerlch et el.. (1973) who noted 100X mortality within 6 month* following administration of 30 ppm PCB (10 ppm each of Aroclor 1242, 1248 and 1254) co adult mink. Feeding of Aroclor 1254 at levela of 5 and 10 ppm In the diet for 4 months led to a dose~ dependent retardation of weight gain of growing female mink. Platonov and Karstad (1973) produced 100X mortality In mink when they fed this epeclee 3.6 ppm Aroclor 1254 In the diet for 105 days. Rhesus monkeys are also very sensitive to PCB* (McNulty, 1975). For example, adult female rhesus monkeys (Hacaca Mulatta) have been fed diets containing 2.5, 5.0 and 25.0 ppm of Aroclor 1248 over periods ranging from 2 months for the higher PCB levela to 1 yeat for the two lower doses (Allen et al., 1974: Baraottl and Allan, 1975). Animals on the 25 ppm dose developed facial edema, alopecia and acne within 1 month, and 1 of 6 animals had expired ae a result of PCB Intoxication 2 months after removal from the diet. The total Intake of PCBs ranged from 250 to 400 mg/anlaal. As was the case with the higher doses, these animals developed anemia, hypoprotelnemla, bone marrow atrophy and severe hypertrophic hyperplastic gastritis. PCB concentrations In subcutaneous adipose tlesus averaged 127 ppm at termination of experimental diet and 34 ppm 6 months later. Surviving animals continued to show clinical signs and lesions of PCB intoxication 2 year* following PCB exposure (Allen, 1975). 109 M0NS 056330 When adult rhesus monkeys were ted Jiets containing 2.5 and 5.0 ppm Aroclor 1246 for 1 year, the females of the group developed periorbital edema, alopecia, erythema and acne from lesions that involved the face and neck within 1 to 2 months (Allen et al. 1974; Allen, 1975). Although males consumed more PCB, they exhibited only moderate periorbital edema and erythema. An abnormal dysplastlc growth pattern of the gastric mucosa was also noted in male rhesus monkeys fed 300 ppm Aroclor 1248 for 3 months (Allen and Norback, 1973). Cygomegalous or squirrel monkeys fed different dietary levels of Kanechlor 400 showed weight loss, palpebral edema, enlargement of the liver and pneumonia. The lowest effective total dose was 300 mg/kg bw given over 20-32 weeks (Nlshlzuml, 1970). A rhesus monkey fed 3 ppm Aroclor 1242 for 8 months died spontaneously and on autopsy was found to possess a gastric lesion manifested by extensive down growth of the epithelium deep into the subserosa and into the muscular wall of the sto mach itself (McNulty, 1975). One monkey fed ?,4,4*-trichlorobiphenyl and another fed 2,4,5,4'5*~pentachlorobiphcnyl at 10 ppm for 90 days were reported clinically well and autopsies showed no positive findings (Oregon State University, 1975; McNulty, 1975). 110 M0NS 056331 Third-litter sows received feed containing Aroclor 12*2 at a concentration of 20 ppm throughout gestation and nursing. Treated sows differed significantly froa control sows In the miober of live pigs farrowed. Treated sows also had more mummi fied fetuses. Performance of live-born pigs was not affected by feeding PCB to the sows during nursing. Pathologic changes in the sovs included hypertrophy of the liver and erosion of the stomach. Slight atrophy of the spleen end thyroid gland were observed In pigs from treated sows (Hansen et al., 1975). Dermal toxicity studies In rabbits of techinlal PCB samples that contain an average of 60X chlorine (Phenoclor DPS, Clophen A60, and Aroclor 1260) as well as fractions containing tetra and pentachlorodlbenxofuran have been recently described by Vos and Beems (1971). PCB-induced skin lesions were hyperplasia end hyperkeratosis of the epidermal and follicular epithelium fol lowing application of 118 mg of the three FCBe (5 times per week for 38 days) in the back skin of adult female New Zealand rabbits. Hlstopathology of thm liver Included centrolobular degeneration, centrolobular liver call atrophy, focal necrosis, and cytoplasmic hyalin degeneration. PCB-lnduced kidney lesions were hydropic degeneration of the convoluted tubules and tubular dilation with the presence of casts. Definitive hyperplasia ana hyperkeratosis of the follicular epithelium of the ear skin were seen after the topical application of fractions of Phenoclor and Clophan, while the fraction from Aroclor caused a minimal hyperplasia and hyper- 111 HONS 056332 keratosis of Che follicular epithelium. Other effects elicited by the dermal application of the PCBs including thymus atrophy and lymphopenia as well as elevated excretion of fecal copro porphyria and protoporphyria. From the response of the back skin and the liver of the rabbit to the three PCB mixtures, and from the response of the ear to the 25X diethyl ether-hexane fractions it was concluded that there were definite quantitative differences in toxicity, at least between the samples used In the above study and prior studies. The extent to which these samples are representative of the normal commercial output has not been established and emphasizes the difficulty in the evaluation of toxicity data of PCBs in which the samples may differ in the amount and nature of toxic impurities. Vos & Notenboom-Rao (1972) compared the toxicity of Aroclor 1260 with a single isomer 2,4,5,2\4*,5'-hexachloroblphenyl In New Zealand rabbits. Dermal applications of a total 120 mg Aroclor 1260 (S times/week for 28 days) resulted In early macro scopic skin lesions. The lesions in a 2,4,5,2*,4*,5'-hexachlorobiphsnyi group of rabblta treated similarly appeared later and were lesa severe. When groups of 5 male mice wore fed dietary levels of 10, 30, 100 and 300 ppm of the PCB isomers 3,4,5,3'4`,5*-hexnchloroblphenyl, 2,4,5,2',4',S'-hexachloroblphenyl and 2,4,6,2',4',6'-hexachlorobiphenyl 112 M0NS 056333 for 4 weeks It was noted that the 3,4,5, 3' ,4' ,3* f isomer of hexachloroblphenyl caused death In nice at a dietary Intake of 10 ppn (2.1 mg/kg bw) within 36-47 days of exposure while consumption of the other two Isomers resulted only In death at a dietary level of 300 ppm (64.3 mg/kg bw). Body weight was reduced at the lowest dose for the 3,4,5,3',4' ,5', Isomer and only at the highest dose for the other two Isomers. Atrophy of the thymus and hepatomegsly was noted at the lowest dose In the mice receiving the 3,4,5,3',4',5', Isomer and only this Isomer produced experimental porphyria. B, Avian subacute toxic effect of PCBs. Chickens fed 10 ppm Arodor 1242, or 100 ppm Aroclor 1254 showed diminished growth but a dietary level of 100 ppm Aroclor 1260 did not produce this effect (Kepllnger, et el., 1971). Symptoms of PCB poisoning In birds consist of tremor, stsxls, ruffling, loss of feathers. Edema of the subcutaneous tissues, and fluid accumulation of the abdominal and thoraxlc cavities are characteristic signs at autopsy. These findings are responsible for this disease being designated chick adema disease. Several outbreaks of chick edema disease caused by PCBs have been reported (Firestone 1973, Xohanowa 1969a, 1969b). Chicks (one day old white Jeghorn cockerels) were fed the following PCB Isomers: 2,3,6,2',3',6'- hexachloroblphenyl 2,4,6,2*,4',6'.hexachloroblphenyl, 2,3,4,2'3',4'-hexachloroblphenyl, 113 HONS 056334 2,3,6,2**3',6', hexachlorobiphenyl and 2,4,3,2*,4',5' hexachloroblphenyl for 21 days at a level of 400 ppm. The Isomer 2,4,5,2',4',5' hexachlorobiphenyl was also fed at the dietary level of 100 ppm. The 3,4,5,3',4',5' Isomer was the most toxic causing death of all chicks within 11 days after onset of exposure. These chicks had chick edema disease, atrophy of the thymus and loss of subcutaneous and visceral adipose tissue. The other Isomers also caused weight loss the 2,4,6,2',4',6' Isomer being the least effective. Liver enlargement was noted for all groups the 2,3,6,2',3* ,6' being the least effective and the 2,4,6,2',4',6* the most effective. The microscopic findings made In the liver were most pronounced In the group fed the 2,4,6,2', 4',6' isomer. (Since the chicks fed the 3,4,5,3*,4',5* isomer died relatively early in the experiment pathological findings in the livers of these chicks are not comparable to thoee made for the other Isomers.) When chicks vsrs fed 5% rice oil which had produced Yusho they developed chick edema In 17 daya. Similar symptoms vara produced when chicks were fed 400 ppm Kaneclor 400 (Goto et al., 1969). C. Mammalian Reproductive Effects of PCBs Oral administration of 0.025 mg/day of Clophen A60 In peanut oil to NMRI-mlce for 10 weeks lengthened the sstrus cycle from 6.6 + 2.5 days to 8.7 + 4.3 days (Orbsrg and Klhlstrom, 1973; Klhlstrom et al., 1973). Females treated analogously for 62 daya 114 MGNS 056335 v- and mated with uncreated malee exhibited a significant decrease in the Implantation rate from 87.OX In the controls to 79.SZ. Male castrated mice were given 0.25 gm PCB (Clophen A 60) In peanut oll/day by stomach cube for 28 days and 70 /pg testosterone propionate dally on days 19-28. The dry weight of the seminal veeclcles after this treatment period was compared to castrated mala mice given tescosterone but not PCB and was found to be slgnlflcanlty reduced. Groupe of 10 NMR 1 female mice were injected subcutaneouely with 50 mg/kg body weight of Clophen A 60 In peanut oil or only the vehicle on the day of parturition and thereafter once weekly for 3 weeks. The 209 offspring were bred when reaching sexual maturity by mating them in pairs as follows: 17 pairs of controls, 19 pairs where the suiles had received Clophen A 60 as a suckling. 23 pairs where the female had received Clophen A 60 as sucklings and 23 pairs were both sexes had received Clophen A 60 as sucklings. The controls had an average of 11.5 offspring per litter while the parents where both mates received PCBS ae sucklings had 8.8 offspring per litter. The resorption race for both groups was ths same but the number of Implantations ware reduced. Reproduction was not affected where only one mate received the PCR (Klhlstrom et al.,1975). Kepllnger et al.,(1972) reported low meting Indices and decreased survival of pups for rats receiving Aroclor 1262 at ^ 100 ppm, and decreased survival of pups receiving Aroclor 1254 115 HONS 056336 at 100 ppm. No reproductive effects were found with Aroclor 1260 at 1, 10, or 100 ppm, or with Aroclor 1242 or 1254 at 1 or 10 ppm. These studies suggest that In mammals reproductive effects decrease with Increasing chlorination. Sherman rats were exposed to polychlorinated biphenyls, Aroclor 1254 and Aroclor 1260. Rats exposed to Aroclor 1254 at dietary levels of 20 ppm or more had fewer pups per litter than the controls in the Fib and F2 generations (Linder et al., 1974). The 100 ppm exposure level of Aroclor 1254 Increased mortality in the Fib offspring and markedly decreased mating performance of the Fib adults. The 500 ppm dietary level of Aroclor 1260 reduced litter size and decreased survival in the FI litters. Dietary levels of 5 ppm Aroclor 1254 and 100 ppm Aroclor 1260 had no effect on reproduction in rats exposed through two generations* Liver weights were Increased in 21-day-old FI male weanlings at the 1 ppm level of Aroclor 1254 and in either sex of FI and F2 weanlings at 5 ppm or higher levels of both Aroclor 1254 and 1260. Histological changes in the liver and Increased liver weights were observed in adult rats exposed to the higher levels. Pregnant rata given Aroclor 1254 at the rate of 100 mg/kg/day on days 7--15 of gestation produced grossly normal litters, but only 10.IX of the pups survived to weaning. Reproduction and pup survival were not affected at dosage rates of 50 mg Aroclor 1254/kg bv/day or 100 tng Aroclor 1260/kg bv/day. 116 M0N$ 05633? Rabbits have been found to be toore sensitive to PCBs than rata in regard to fetotoxic and reproductive effects (Vllleneuve et al., 1971). In rabbits, no effects were reported at dosages of 6.25 or 10 mg Aroclor 1256 /kg/day, but at dosages of 12.5 to 50 mg/kg/dey, fetotoxic effects were noted. Abortions, maternal death and stillboms occurred, but no consistent skeletal abnor malities vers found. Ringer et al., (1972) found that 1 ppm of Aroclor 1256 caused a alight reduction In reproductive aucceaa in mink, but at 5 ppm Aroclor 1256 complete reproductive failure wee noted. In the same study 12 mink were fed a diet containing 30% Coho Salmon from Lake Michigan. Hone of the mink whelped while 11 of 12 controls whelped. Female adult rhesus monkeys fed Aroclor 1268 at a level of 25 ppm for 2 months noted at beginning of the 5th month experienced either fetal resorption or abortion during their second month of pregnancy (evidenced by regression In uterine site and reestablishment of menstruation (Allen et el., 1976). A third animal carried her fetue to term. This infant's weight wee considerably lover than the average rhesus Infant (375g vs 566 + lOlg) and had 25 A>g PCr-/g/(ppm) of adrenal and adlpost tlaaue. Adult female rtv.jue monkeys fed 2.5 and 3.0 ppm Aroclor 1268 wera bred to control malea following the establishment of a relatively steady tissue level of PCB at 6 months. Following 3 mating, 12.5% of the 5 ppm group and 37.5% of the 2.5 ppm group was pregnant 117 HONS 056338 compared to 90Z in the control group (Allen, 1975). The conception rate of females bred to PCB-fed males was equally as great as that In females bred to control males. I). Mammalian Sub-Acute and Reproductive Effects of PBBs Male Sprague Dawley rats maintained on diets containing 0, 0.1, 0.01 or IX octobromobiphenyl for 30 days did not exhibit overt toxicity during this period. Gross pathological studies revealed enlarged livers and some kidney changes (petechial hemorrhage, enlargement), while hlstopathologlcal changes in the IX group Included liver lesions consisting of centrilobular cytoplasmic enlargement and vaculation and kidney lesions conslating of hyaline degenerative cytoplasmic changes (Aftosmls et al., 1972). Thyroid hyperplasia was observed in all rats In all test groups. Rats were fed 100 and 1000 ppm octabromoblphenyl in their diets and then a number were sacrificed after 2 and 4 weeks of feeding and after 2,6 and 16 weeks of recovery on an octabromobiphenyl-free diet. The hepatocytea were markedly enlarged in rata fed 100 ppm and 1000 ppm of the PBB after 2 weeks of treatment. Histopathologic changea were characterized by hepatocellular hyper trophy with the pathologic changea localized mostly In the centralobtilar zone (Aftomis et al,, 1972b). The cytoplasmic Inclusions were more pronounced in the livers of rats fad 1000 ppm than 100 ppm of the PBB. There was some evidence of recovery since the Livers of the 100 ppm level animals returned almost to normal 2 weeks after 118 MGNS 056339 withdrawal of the compound. Preliminary 28-day feeding studies In rats with octabroooblphenyl at 100 and 1000 ppm demonstrated liver enlargement, hepatocellular alterations with cytoplasmic inclusions and accumulative effects of bromine In the fat, liver and muscle (Aftosmls et al., 1972). Norris et al., (1974) demonstrated hematological changes, liver enlargement and liver and kidney lesions at all levels of octabromoblphenyl (10, 000, 1000 and 100 ppm) In the diet of rats. When three groups of prlmagravld rats fed diets containing 100, 1000 or 10,000 ppm of octabromoblphenyl (BB-8) from day 6 through 15 of pregnancy and the pups delivered on day 20, anasarca was observed in one fetus at each of the two highest levels and gestroschlsls was observed In another fetus at each of these levsls. No other gross effects were observed In either the mother* or the pups and dose-related levsls of bromine were found in the fetuses (Aftomls et al., 1972a). When 1 g/kg of octobromobiphenyl was applied in corn oil under occuluded conditions to rabbits for 5 hrs/day for 10 days over a 2-week period, liver enlargement vaa found whllt at 0,lg/kg no enlargmant was found (Aftosmls, at si., 1972b). Hexabromoblphenyl (Flremastsr BP-6) fed to pregnant SpragueDawley rats and Swiss/ICR mice (100 and 1000 ppm) resulted in a dose dependent decrease in mean fetal weight. (Number of pregnant 119 HONS 056340 mice: 16 controls, 9 received 100 ppm and 12 received 1000 ppm. Number of pregnant rata: 12 controls, 7 received 100 ppm and 6 received 1000 ppm). Exencephaly was produced in the offspring of the mice that received both 100 (3/121)and 1,000 ppm (2/174 dosages while cleft palate (4/87) and defective kidneys (2/87) were noted in offspring at the 1,000 ppm level. Nonpregnant mice fed 1,000 ppm PBBs for 11 days showed marked Increase in liver else and weight (Corbett et al., 1975). Cows from the Halbert herd received feed containing PBB at a level of 2914 ppm at a consumption rate of approximately 15 lbs/head/day (200 cows) or 8 lbs/head/day (200 cows) for a period of approximately 16 days. Early signs of toxicity included anorexia, decreased milk production, increased frequency of urination, lacrlmatlon, some lameness, and shrinking of the udder of recently freshened cows. Upon removal of the contaminated feed appetite Improved. Cows bred 4-6 weeks prior to onset came back in heat suggesting early resorptions. Later signs exhibited by the cows included hematomas, abscesses, weight loss particularly In high producing cows, abnormal hoof growth, rough hair coat, alopecia, thickening of skin, dystocia, lack "tf udder development at freshening, metritis, and neg lglble milk production in cows freshening 3-4 months post-exposure. Cows also exhibited a lack of wound healing, general weakness and were highly susceptible to stress. Six months post-exposure non-lsctstlng cows showed depression of 120 HONS 056341 V appetite, prepartua weakness, failed to develop signs of labor and died without calving. Grose pathology included liver and renal degeneration, and hematomas and abscesses in the peritoneal and thoracic cavities. Twelve 6 to 18 month-old heifers and bulls were offered the contaminated feed ad libitum. Five of the anlmale were dead within 6 weeks. Younger animals became prostrate, went through a short period of coma and died. Signs and lesions Included testicular atrophy, abdominal adhesions, and massive liver abscesses. At S months only 2 animals remained alive. Theee became anorexic, would consume only milk and developed signs of hyperkeratosis over the entire body (Jackson and Halbert, 1974; Uelbom, 1975). The health status of 16 herds of dairy cattle exposed to low levels of polybromlnated biphenyl (PBB) was compared with that of IS control herds. Milk production of the contaminated herds was not slgniflcangly changed In 1972, 1973, and 1974, and was not significantly dlfferant from that of control herds in the same years. Mortality of adult cows and calves, the percentages of cows culled from the herds because of old age and low production, disease, or sterility, and the general health conditions were similar in the two groups. Strum concentrations of calcium, glucose, and cholesterol in contaminated herds were significantly different from those of the control herds, but the relationship to PBB exposure needs further investigation (Mercer et al., 1976, In Press). 121 ONS 056342 A study consisting of an investigation of records and herd history of herds accidentally exposed to PBBs was also done. Significant findings Included a decrease in production of 20 to SOX; severe weight loss at calving time; sterility; decreased growth rate in young animals; evidence of soreness and stiffness; poor response to therapy of common diseases; prolonged wound healing; abnormal hoof growth; calf losses; and malformed calves. (Demlng, 1975). Polybromlnated biphenyl was administered to four cows at a dose rate of 10 mg/head/day for 60 days. Although the study was not designed as a toxicology study, retrospective evaluation of the data provided no evidence of clinical problems among teat animals during the period of exposure to PBB nor for a year thereafter (Fries and Marrow, 1975). E. Chlorinated Dibengofurana - Mammalian Toxicity It is presently assumed because of our experience with the chlorinated dlbenxodloxlns, that the 2,3,7,8-tetrachlorodlbenzofuran is the moat toxic compound of this group of chemicals, while the dlbensofursn Itself or the octadlbenzofuran have little Inherent toxicity on an acute basis (Kimbrough, 1974). Bau. r ot al, in 1961 demonstrated the toxicity of mixtures of trl and tetrachlorodlbenzofurans. A single dose of 1 mg or 0,5 mg/kgbody weight given orally to rabbits caused severe and often fatal liver necrosis. Application of this material to the rabbit 122 MQNS 056343 ear cauaad severe hyperkeratosis at the application site. The single oral LDjq for tetrachlorodibensofuran for guinea plga la between 5 and 10 /ug/kg. Symptoms of toxicity were severe weight loss, atrophy of the thymus and spleen. Hemorrhages were observed in the adrenals, urinary bladder and single cell necrosis was noted In the liver. C. D. Strain rats given as much as 1000 /ug/kg bw of the TCDF failed to show any symptoms of toxicity and microscopic examination of their tissues did not reveal any abnormalities. Similarly oral single doees of up to 6000 /ug/kg body weight of TCDF failed to elicit any toxic effect In CS7B1 mice. A subcutaneous dose of 6000 /ug/kg bw given to these mice produced weight loss, hepatomegaly and atrophy of the thumus but no fatalities (Moore et al., 1976). While these findings Indicate that In mice TCDF la 30 fold lesa toxic than TCDD, the overall estimated difference la toxicity between TCDD and TCDF Is about tenfold (Bauer et al., 1961; Kimbrough, 1976; J. C. Vos, personal communication). F. Chlorinated Dlbensofurans - Avian Toxicity When 1 day old chicks were started on dally oral doses of 5 /ug/kg TCDF dally most of them died within 11.3 days. One of 6 chicks died g.,en dally oral doses of 1 /ug/kg bw. Symptoms of toxicity at the low as wall as the high doses consisted primarily of weight loss, decreased food consumption and general unthrlfty- ness. At autopsy the most striking findings were subcutaneous edema, ascites and hydropericardium and atrophy of the thymue. 123 HONS 056346 An Inflammatory exudate was noticed on microscopic examination In the lung and the pericardial surface of the heart showed Inflammatory changes. Congestion and hemorrhage into the gastrointestinal tract was also observed (Moore et al., 1976). G. Bromlnated Dlbenzofurana - Mammalian Toxicity When dally doeea of A/ug/rabbie of 2,3,7,8~tetrabromodlbenzo~ furans were applied to the rabbit ear for 5 days giving a total dose of 20yug/rabblt or about 5~6/ug/kg bw, the rabbits (A) developed hyperkeratosis of the treated ear and areas of liver cell necrosis (Kimbrough, personal coomunicatlon). Mo other information la presently available on the toxicity of bromlnated dibenzofurane. H. Longterm Toxicity Including Tumorogenesis Chlorinated biphenyls Mice When male BALB c/J mice were fed Aroclor 125A for 11 months at a dietary level of 0 or 300 ppm (A9.8 mg/kg body, weight) 22 of 30 mice In the experimental group and 2A of 50 In the control group survived* The livers of the teat animals were markedly increased In else, and showed a qualitative increase in porphyrin. A total of 9 experimental mice shoved 10 neoplastic nodules (hematomas, hyperplastic nodules) in their liver ranging in site from 0.1 - 0.5 cm In diameter. Other morphological changes observed in the livers of the experimental group including plcomorphltm. 12A HONS 056345 areas of necrosis, and foci of adenoflbroala. (Klabrough and Llndar, 1974). Nalthar tumors nor tha othar morphological changed vece noted In the controls. Only one of 24 surviving male KALB c/J mica fed Aroclor 12S4 for 6 months folloved by a control diet for 5 months had a hepatoma (neoplastic nodula) measuring 0.3 cm In diameter. Ito at al., (1973) fed male dd mica 500, 2S0t 100 and 0 ppm Kanechlor 500, 400 and 300 In the diet respectively. After one year, hepatocellular carcinomas were observed In 5/12 mica fad Kanechlor 500 at a dietary level of 500 ppm, tha othar 7 mice In this group had nodular hyperplasia (neoplastic nodules?). No metsstases ware observed. Feeding a, B, or X hexachlorocyclohexana (BHC) isomers with or without Kanechlor 500 at a dietary level of 250 ppm to mice for 24 weeks did not produce any tumors In the liver of animals receiving only the FCB (Ito et al., 1973). Liver tumors were observed in mica receiving 250 ppm a BHC alone or 250 ppm FCB and either 100 or 50 ppm a BHC or 250 or 100 ppm B BHC. Thus FCBs enhanced the tumor development of liver tumors of a or B BHC Isomers. Rats Dietary exposure of mala Sherman strain rats to levels of 500 ppm Aroclor 1254 (36.4 mg/kg body weight) for 6 months resulted In pronounced lipid accumulation In tha liver which persisted for a 10 months observation period following tha discontinuation of 12'. HONS 056346 the dietary exposure to PCBe. AdenofIbrosls (cholangloflbrosis) which was observed in the livers of rats ingesting the PCB lor 6 months was still present 10 months later. Whether this lesion is persistent could not definitely be determined since high levels of the higher chlorinated biphenyl Isomers were still present In adipose tissue and liver (Kimbrough, et al., 1973). Adenoflbrosls is a focal proliferation of glandular epithelium forming ducts often surrounded by extensive fibrosis. This lesion usually occurs concomitantly in rat livers with hepato cellular carcinomas and has been produced by many known hepatocarclnogens. It was first described by Edwards and White (1941). The extremely long retention of some PCB isomers in rats was again noted when male Sherman strain rats were fed 100 ppm Aroclor 1254 for 6 months and then allowed to recover for 16 months. 4.4 ppm of PCB derived material was present In the liver and 152 ppm in adipose tissue on s wet weight basis (Kimbrough, 1976). Continued dietary exposurs to commercial PCB mixtures In mammals results in their storage in adipose tissue and over an extended period high levels may be attained (Curley, et al., 1971; Burse, et al.t 1974), Blood levels are more a function of the dose, ths rate of metabolism, availability of binding sites In blood, chemical characteristics of the compound given, rather than the amount stored in adipose tissue (Smrek, et al., in preparation). I 2f> MONS 056347 la Another study, 200 experimental female Sherman strain rats were fed 100 mg/kg Aroclor 1260 In the diet (11.6 - 4.3 sg/kg body weight) for approximately 21 months and 200 female rats were kept as controls. A total of 184 experimental rate and 173 control* survived to sacrifice at 23 months. .Hepatocellular carcinomas were observed In 26/184 experimental rats and In 1/173 controls. None of the controls but 146/184 experimental rata had neoplastic nodules within the liver (hyperplastic nodules). Areas of hepatocellular alteration were noted In 28/173 controle and 182/184 experimental rats. The incidence of tumors In other organs was not altered by PCB Ingestion and metatasee from the liver tumors wars not observed (Kimbrough at si., 1975). AdenofIbrosls of the liver wee also noted In this study. In reporting these liver tumore the classi fication developed In a rat liver tumor workshop was followed (Squire and LeVltt, 1975). Liver tumors (multiple adenomatous nodules) vers also induced in 6/10 female Donryu rate with Kanechlor 400 but not in male rite. The dietary exposure varied throughout the study from 38-616 ppm (Kimura and Baba, 1973). The induction of liver tumors by known carcinogens In Sprague Dsvley rate such as 3'methyl-4*dlm*chylamlnoazobenzene, N-2-fluorenyl acetamide and dlethylnitrosemlne wee inhibited by Kanechlor 500 (Maklura et al.,1974). Groups of 50 male and female Charles River rate were fed dietary levels of 0, 1, 10 and 100 ppm of Aroclor 1242, 1254, and 1260 respectlvsly (Calandra, 1976), At 3, 6, and 12 months after 127 HONS 0563*8 onset of the experiment, 5 rats per each group and sex were sacrificed. The surviving rats were sacriflcied 24 months after onset of the experiment. The only body weight depression noted wee observed in the female ruts fed Aroclor 1254. The author reported that important histopathologic changes were only noted In the rats fed the different Aroclors for 24 months. The liver weights were Increased in the male and female rats fed 100 ppm of Aroclor 1260 and 1254 and only In females fed Aroclor 1242 at a dietary level of 100 ppm. It was noted that of a total of 20 surviving male and female rats (10 male and 10 female rats) fed 100 ppm Aroclor 1242 In the diet for 24 months 8/20 rsts showed liver lesions called nodular hyper plasia, 2/20 had hepatomas and 1/20 had a cholanglohepatoma. A total of 27 male and female rats (13 male and 14 female rats) survived to 24 months In the groups of rat9 fed 100 ppm Aroclor 1254 of these 13/27 hsd nodular hyperplasia of the liver 4/27 showed hepatomas and 2/27 showed cholangiohepatomas. Similar findings were made in the total 27 surviving male and female rats fsd 100 ppm Aroclor 1260 for 24 months. In these groups 7/27 male and female rats had nodular hyperplasia of the liver. 5/27 had hepatom.<s, and 2/27 had cholangiohepatomas. Croups of 4 male and 4 female dogs were fed dietary levels of 0, 1, 10, and 100 ppm Aroclor 1242, 1254, and 1260 respectively for 2 years, 1 female fed 100 ppm Aroclor 1242, 1 male fed 10 ppm Aroclor 1242, 1 female dog fed 100 ppm Aroclor 1254, 1 female 128 MONS 056349 V fed 10 ppm and 1 male and 1 female ted 100 ppm Arm'lor I2t0 died. At the dietary level of 100 ppm of either Aroclor 1234 or Aroclor 1260 the body weight gain was slightly decreased for both sexes. Aroclor 1260 also reduced thf body weight gain in the females at the dietary level of 10 ppm. In the beagles fed Aroclor 1260 at a dietary level of 100 ppm, the liver weight was Increased and the serum alkaline phosphatase was elevated. All but 1 dog that died had either chronic peritlonitis or chronic pneumonitis. Many of the dogs fed 10 or 100 ppm of different PCBs sheeted chronic Inflammation of a variety of organs. Male dogs fed Aroclor 1234 showed diffuse Hyperplasia of ths interstitial cells of the testes and aspermatogenesis (Monsanto, 1971). Pecachloroblphenyl No long term studies are available. Polybromlnated Biphenyls No long term studies are available. Chlorinated Dlbensofurana and Chlorinated Naphthalenes No long term studies are available. Immunosuppressive Effects of PCB and PBB Polychlorinated biphenyls (PCB) have been Implicated as immunosuppressants* Friend and Trainer (1970) found s decreased . resistance to duck hepatitis virus In PCB exposed ducklings. 129 MONS 056350 PCB has been shown to produce lymphoid atrophy In rabbits (Street and Sharma, 1975; Vos and Beems, 1971), chickens (Flick et al., 1975; Vos and Koeman, 1970) and guinea pigs (Vos and DeRolj, 1972; Vos and van Cenderen, 1973). In addition, PCBs were found to suppress humoral Immune responses to several antigens In rabbits (Roller and Thigpen, 1973, Street and Sharma, 1975) and guinea pigs (Vos and DeRolj, 1972, Vos and van Cenderen, 1973) and to suppress cell mediated Immune reactions In guinea pigs (Vos and van Cenderen, 1973). In all of these studies the PCBs studied have been commercial mixtures; the role of contaminants such as chlorinated dibenzofurans needs to be considered. The effect of HBfi on the immune system was studied in chickens and guinea pigs (Vos and van Cenderen). In chickens the atrophy of the lymphoid tissue was observed in bursal follicles and in the spleen. It suppressed the humeral immune response in guinea pigs. MUTAGENICITY AND TERATOLOGY Polychlorinated Biphenyls The mutagenicity and cytotoxicity of PCBs. Aroclor 1242 and 1254, were studied by Gren er al., 1975, a, b. The lack of mutagenic activity as measured by the dominant lethal test was reported for male Osborne-Mendel rats subjected to four different dosage regiment: (1) single oral intubations of 625, 1250 or 2500 mg/kg (for Aroclor 1242); (2) five daily doses of 125 or 250 mg/kg (for Aroclor 1242), 75, 150 or 300 mg/kg (for Aroclor 1 JO M0NS 056351 1254); (3) five daily doses of Aroclor 1254 at 150 mg/kg, then starved overnight prior to mating; (4) chronic feeding for 70 days at a dietary level of 25 or 100 ppm prior to mating (Green et al., 1975a). In a second study, bone marrow and spermatogonial cells were cytogenetically investigated from Aroclor-treated Osborne-Mendel rats. Aroclor 1242 was given orally at single doses of 1250, 2500, or 5000 mg/kg or as four dally doses of 500 mg/kg/day; Aroclor 1254 was administered orally as five dally doses of 75, 150, or 300 mg/kg/day. No statistically significant Increases in chromosomal aberatlons were found for the Aroclor-treated rats* As indicated by Green et el , (1975 b), while these studies were negative in regard to chromosomal mutations, they do not entirely rule out the possibility that PCBs may Induce point mutatlona. To date, there are no known reports In the literature concerning the Induction of point mutations by PCBs in laboratory model systems. The observations of Nilsson and Ravel, 1974, in Drosophila where no chromosomal breakage was found among Clophen A30- or A50-treated flies, are fully consistent with those of Creeo et al., (1975, a, b). TERATOLOGY Female NHRI nice, orally administered 0.025 mg/day of Clophen 60, showed evidence of an Increased estrus cycle from 6.6 + 2.5 days to 6.7 4*3 days and a decreased ova Implantation rate from 87.0 to 79.5X (Orberg and Klhlstrtfm, 1973). Kihletrb'm et al., (1974 a), also demonstratad a decreasa in the frequency of implanted ova 131 HONS 056352 among young animals nursed on milk containing PCB (Clophen 60). Both the male and the female contributed co the magnitude of rho reduction of implantations, the mechanism by which exposure ot the developing male to milk containing PCB leads to a reduction in the precent of ova implanted is unknown but need not necessarily involve dominant lethal events. No reports of frank terata were found, but a number of studies indicated the facility of PCBs for placental transport. Grant et al., (1971 b), showed that the concentration of PCB in feral tissue from oral administration of Aroclor 1254 was dose-dependent and that accumulation occurred in fetal liver where higher concentra tions were found than In maternal liver. Placental transport of PCB has been reported for the rabbit, ret, mouse, and cow (Grant el: al., 1971; Platonov and Chen, 1972; Berlin et al., 1975; Melvas and Brandt* 1973; Brandt and Ullberg, 1973). Evidence of placental transfer was also observed among Yusho patients. Wnile PCB has no known or clearly defined teratogenic effect in mammals, their easy passage across the placenta suggests the possibility of potential for some form of fetal toxicity. Chlorinated Dibensofurans No information available. Bromlnated Biphenyls 15 pregnant rats were force fed 100 mg hcxabromobiphenyl/kg 132 i MONS 056353 body weight on days 6, 8, 10, 12, 1A and 16 of pregnancy in corn oil. 1A control rata were given corn oil. The pupa were obtained by cesarian on day 19 of pregnancy. No cerate were observed and no difference between control and experimental rata vaa noted In any of the other parameters studied. 5 control and 6 PBB treated animals were treated with colchicine and bone marrow was studied cytogenetically. Animals treated with PBB and colchicine had higher metaphaae and mitotic indices than non-treated animals. Chromosome aberrations were not noted (Plscor and Werts, 1976). Toxicity of Chlorinated Naphthalenes The following information is derived from e review publlehed by the EPA, namely Environmental Hazard Assessment Report 'Chlorinated Naphthalenes', EPA-560/8-75-001*. For lltereture references please refer to this report. Animal Studies The following tables summarize the evaileble studies in laboratory animals. Three routee of exposure ere reported. *Thie document Is available through the National Technical Information Service, Springfield, VA 22151. 'Preliminary Environmental Hazard Assessment of Chlorlneted Naphthalenes, Silicones, Fluorocarbons, Ben&enepolycarboxylste* and Chlorophenols'. (NTIS accession number PB-238-07A/AS) 133 MONS 056354 Compound MonoDl- TrlTrlTri/Tetra Penta Peata/Hexa- Penta/Sexa Penta- HexaBexaHexaBepcachloroOctaOcta Species Rat Mouse Rat Rabbit Rat Rat Rat Guinea pig Rat Rat Rat Rabbit Rat TABLE 9 (a) Dietary Exposure Dose Level Days Exposed None reported 5g/Kg feed 15 2.5 ng/day 300 ag/day 15 ag/Kg body vt. 50 ag day 20 9-136 60 63 300 ag/day 33 100 ag/day 55 2.5 ag/Kg In oil 48 20 ag/Kg 63 ag/Kg 200 ag/ICg 1g 0.5, 2 or 5g/Kg 84 84 84 None reported Single dose 22 Effect Reported Increase in liver weight, growth impaired, rough coat. None. Fatty infiltration of liver. None. Jaundice/fatty degeneration of liver. Death with yellow livers and extreme fatty degeneration. Death with yellow livers and extreme fatty degeneration. Death-Severe weight loss fatty degeneration of liver. Weight loss. Weight loss. Death. Death in seven days. Decrease in liver but not plasma Vit. A. HONS 0 5 6 3 5 5 ( TABLE 10 (b) Inhalation Studies Compound TriTriTri- Penta- Specles Bat Bat Rat Rat Dose Level 0.05-0.2 -g/L 1.31 ag/L 10.98 mg/L 1.16 ag/L Days Exposed Effect Reported 2 hr/day for 20 days 16hr/day for 134 days 16hr/day for 102 days 16hr/day for 52 days No toxic effect. No toxic effect. Slight liver discoloration, 52 rats shoved increased fatty degeneration Jaundice, enlarged liver and 692 fatality. No studies have been reported for the other chlorinated naphthalenes. 9SC9S0 SNOW (c) Denial Studies Coapound Mono-- Mono- TriPentaPenta- Species Babbit(ear) Babbit(ear) Bat(skin) Mice (skin) Swine (skin) Babblt(ear) Babbit(ear) Cone Applied 90 *g/g Acetone 500 Bg/g Acetone Not given Not given 60 g/Lx3L 30 mg/g acetone/day 30 ag/g acetone Days of Application 5-7 5-7 2 hr/day/40-60 days 2 hr/day/40-60 days 6 days/veek-4 weeks 5 5 Effect Mild reddening. Severe reddening, no decrease None reported None reported Slight hyperkeratosis. Mild dermatitis, hair Decrease In sebaceous glands TOXICITY IN BIRDS Turkey Poults Penta/Kexachloronaphthalenea in the diet. (a) 5 ppm for 40 days caused 23X reduction in weight gain with 6.5% mortality. (b) 100 ppm - all poults died within 33 days. Histo pathologic examination showed enlarged and darkened livers. The LC^q was 20 ppm with an average decrease In weight of 51%. Chickens Penta/Hexachloronapthalene in che diet. (a) No effects reported at dietary levels of 4 and 20 ppm (35 days). (b) 100 ppm prevented egg production (35 days). (c)-2550 ppm causad 100X mortality (14 days). Effects reported Included enlarged fibrous livers, lack of feather pigmentation and pericardial and peritoneal edema. A fourfold increase In Vitamin A in the diet dacreaaed toxic effects. 136 MONS 056357 TOXICITY IN MMJMTIC ANJHAIS Ingestion 1b the most Important route for exposure of domestic animals. Cattle Highly chlorinated naphthalene poisoning In cattle Is known as bovine hyperkeratosis or X-dlsease. Detailed Information on toxic doses la lacking. Penta/hexa mixture at a dose level of S.5 mg/Kg body wt (orally) for 5 days caused a sharp drop In plasma Vlt. A by the end of the third day and depressed plasma Vlt. A for over 30 days. A single oral dose of 1 mg/Kg body wt of hexachloronaphthalene caused mortality within two weeks. The first effect of chloronaphthalene poisoning is an interference with the blotreneformatlon of carotene to Vlt. A. The Vlt. A. deficiency Is quickly followed by lnflanaatlon of the oral mucosa, lacrlmatlon, excessive salivation and Irregular food conaumptlon. Grosa physical effects that develop during the course of disease Include a general thickening of the skin caused by over development of the skin's hairy layer with loss of hair (hyperkeretoala). There may be degeneration or Irregular growth of the horns. Continued exposure results In anemia, dehydration, loss of weight, fever and death. Thera may be e< ere liver damage. Swine No toxic effects were observed at levels of 100 mg/Kg 137 MOMS 056358 body vc of the chlorinated naphthalenes (degree of chlorination was not stated). 150 mg/kg body vt caused a marked depression of plasma Vit. A, and death occurred at a dose level of 198 mg/ Kg body wt. Sheep Toxic effects are observed at doses of chlorinated napthnlene 10 X that required to produce toxicity in cattle. Sheep do not show cutaneous hyperkeratosis or an excessive drop in plasma Vlt. A as observed In cattle. Effects reported include nasal discharge, weakness, loss of weight, loss of appetite, ascites, necrosis and cirrohsls of the liver and cardiovascular effects. Toxicity in Man The most important route for mans occupational exposure to chlorinated naphthalene is via the inhalation route. Dermal absorption route may also occur. Penta/hexachloronaphthalene at a concentration of 30 mg/g acetone, applied to the backs of human volunteers ior a period of 9lx weeks caused typical chlorucne. Dally topical application of mono-dlchlorlnated naphthalene to the human ear In mineral oil suspension for 30 days caused no observable effect. Two clinically distinct but oftern concurrent syndromes have been described, namely, liver necrosis, and chloracne I 3H hons 056359 with Itching* A number of deaths involving acute atrophy oi the liver have been reported; with the course of the disease similar to other forms of severe liver damage. Autopsy of fatally exposed workers show severe yellow atrophy of the liver. Definite data relating to dose response effects are In general lacking. However, two dermal studies with man have been reported. SUMMARY The degree of toxicity of chlorinated napthalenes Increases with the degree of chlorination. The higher chlorinated naphthalenes (especially hexa and penta) have been associated with chloracns and liver damage In man. Dose response effects have not been estab lished. Cattle are more susceptible to the effects of chlorinated naphthalene chan other domestic animals. Tha effects observed Include development of Vlt* A deficiency, hyperkeratoele and liver damage* 239 HONS 056360 HUMAN EXPOSURE TO POLYCHLORINATED BIPHENYLS 1. Introduction Although the polychlorinated biphenyls (PCBs) have been used in a wide variety of industrial applications in the U.S. from 1930 until 1970, when their distribution was voluntarily restricted to closed systems, there is scant Information on human exposure to adverse human health effects in the U.S. In the U.S., minimal human exposure of the population to PCBs is Limited to food, air and water, while significant human exposure appears to be limited to sports fishermen consuming fresh water fish from contaminated streams and lakes; and to occupational exposure in Industrial workers. II. Dietary Exposure to PCBs The May 1972 Report of the Interdepartmental Task Force on PCBs 8utranarlzed the findings in food sampling programs con ducted by the Food and Drug Administration and the Department of Agriculture. Jelinek et al., (i 975), have provided an update to that summary and evaluated trends which is presented below. The 1972 Task Force report outlined three broad, sources of food contamination. These are restated here in order to show which sources have been effectively controlled and which sources appar ently persist, as reflected by results obtained In continued food sampling. HONS 056361 1. Environmental contamination - background levela of PCtte in fish from lakes and streams. 2. Industrial accidents - Isolated incidents Involving direct leakage and spillage or contact of PC8 fluids and other PCB containing materials on animal feeds, feed ingredients or food. 3. Food packaging materials - PCB migration to foods packaged in PCB contaminated paper products. Ae discussed In the 1972 Task Force report, there were a num ber of other positive findings In foods which could not ba traced to one of these broad sources prior to 1972 and tha source of such occasional findings remain speculative. However, In more recent years, essentially all positive findings of PCBs in the food sup ply have bean attributable to one or more of these three sources. Jelinek and Cornelluesen (1975) reported that for the 1969-1975 period, there has been a significant decrease in PCB levels in all foods with the exception of fish, where no particular trend has been noted. From that observation they concluded that procedure* instituted to exclude the use of PCBs In "open" applications have bean effective, but that more needs to be done to prevent their entry Into the aequatlc environment. Jelinek and Comelluasen (1975) presented date on FDA's sampling program showing a very conclusive decline In the FYs 1973-1975 HI MOWS 056362 period in Lernw 01 occurrence rale of I'Clis and maximum ioveiy found in milk, eggs, cheese, animal foods and components, processed fruits, and baby foods. Except for fish, occurrence rates in FY'75 had dropped to zero or less than 1% for all categories, and there were no findings in excess of the existing temporary tolerances. Similar data were presented covering USDA's sampling of meat and poultry which also showed a decline, to 0.3% positive and only 0.6% above the temporary tolerance. Jelinek and Corneliussen (1975) ilso reviewed I'DA Total Diet Study findings (FY'71-75) which showed that all food classes of the total diet have declined to no PCB occurrence and no calculable dally Intake of PCBs, except In the meat-fish-poultry composites. About 40% of these composites continue to contain detectable PCBs, although only traces have been detected in the latter years. The limit of quantitative detection in the Total Diet Study is about 0.05 ppm; positive findings below that level are generally reported as traces since they are not reliably quantitated. Occurrence of PCBs ended in 1973 in all other categories, notably in dairy product composites and the composites of grain and cereal products. This probably reflects the control measures effected on animal feeds and on the control of ingredients in recycled paper t >d for food packaging. What remains, baaed on the total diet study, is a continued low level occurrence (about 40% positive and only at trace levels) in the ment-fish-poultry HONS 056363 composites. The fact that levels in these composite* have declined to only trace* further supports the inference that the meet end poultry and eggs no longer contain detectable PCBs and that the low level findings are primarily due to the fish in those composite*. This would imply that the PCB levels in the diet may have "bottomed out" and may remain static until such time as there is a change in the levels for fish. Estimates of dietary Intake of PCBs bassd on the Total Diet Study must be taken as gross estimates because only traces are currently found and some type of estimated value must be assigned to those traces to calculate microgram per day Intakes of PCBs. Jellnek and Coraelluaaea used a set of consistent assumptions for those traces In computing ths dally Intakes for the FY-1971-75 period. Their calculations shoved e steady decline from IS meg per day down to 8.7 veg/day, with all of this coming from the meat-flshpoultry group In FY'74-75 (minor and declining portions attributable to other food cleeses in earlier year*.) Review of this Information Implies that, although the verlous assumptions for trace residue reportings and varying food consump tion data hava a significant effect on dally intake calculations, an overall estimate In the order of 5-X> aeg/day PCB dietary intake of PCBs for th gensrsl public seems ressonable. This flgurs would not bs applicable to diets which may Includt regular consumption of fish from certain location* with high PCB levala. 141 HONS 056364 III. PCB Residues in Fish Kevluw or the data gathered by various agencies on fish resi dues Indicates a serious Incompatablllty of sampling programs for obtaining data which reliably define trends in PCB levels. The Food and Drug Administration's primary concern is the levels which exist in the edible portion of commercially imported fish which are marketed interstate. Therefore, In moat cases, the heads, entrails, skin and fins are excluded. Furthermore, FDA sampling has not Included fish which are caught and consumed locally (freshwater fish) and the sampling has dealt with the most impor tant consumption species which are primarily of marine origin and which generally do not contain PCB levels as high as freshwater species from certain locations. In order to determine the human exposure to PCBs through dietary fish, one must know the levels of PCBs residues In the edible portions of fish, and the amounts of the various kinds of fish consumed by the population at large, and by special subpopula tions. During the past few weeks, information has been compiled by National Marine Fisheries Service-HOAA on the PCBs levels of these residues in marine sad freshwater fish and shellfish, and on dietary habits. Tabla 11 lists the 20 most important types of fish eaten in the U.S. today, and the mean daily amount of each type eaten by 144 HONS 056365 TABLE 11 v*.- . Fih and Shellfish Consumption in ths United States (September 1973-Auguat 1974) Total Tuna (mainly canntd) Unclaatlflad* (mainly braadad, incl. flah atlcka) Shrimp Ocaan Parch* Floundar Clana Craba/Lobatara Salmon Oyatara/Scallopa Trout** Cod* Baaa** Catfiah** Haddock* Pollock* Harrlng/Smelt/ Sardlnaa Pika** Halibut* Snappar Whiting All othar claaalflad Amount (mill.) Rank lba/vr) 2957. 1 634 2 542. Percent of total by vt. .100 21.4 16.4 Nuaibar of Actual Uaa (millions) 197. 130. 66. 3 301. 4 149. 5 144. 6 113. 7 110. 8 101. 9 88. 9 88. 11 78. 12 73. 12 73. 12 73. 15 60. 16 54. 17 35. 16 32. 18 32. 20 25. 152. 10.2 5.0 4.9 3.6 3.7 3.4 3.0 3.0 2.7 2.5 2.5 2.5 2.0 1.8 1.2 1.1 1.1 0.9 5.1 45. 19. 21. 16. 13. 19. 14. 9. 12. 7.6 7.5 .11 .11 10. 2.5 5.0 4.3 3.2 n.a. * Mainly inporta ** Fraah water H*4d mount par usar (Arana/day 16.7 6.1 10.0 8.3 9.7 6.6 7.6 10.6 6.7 7.8 12.3 8.1 12.0 12.1 8.6 6.6 6.7 17.4 8.0 9.3 9.7 n.a. HONS 056366 che subpopulations of actual users. The Table Includes all dietary fish, from sport fishing and from domestic and Imported commer cial fishing. These 20 items comprise 95Z of the fish products eaten. The results are taken from information in a recent Seafood Consumption Study by the National Purchase Diary of Schaumburg, Illinois, sponsored by the Tuna Research Foundation, and National Marina Fisheries Service data on the amount of fish and shellfish processed and distributed in the U.S. Several items in the Teble are worth noting for purposes of the present analysis. (1) Although ninety-three percent of the U.S. population (197 million) eat fish, the average annual per capital consumption of fish Is email: 1S.0 lbs/year--fish eater; (2) a large "Unclassified" fish fraction exists in the U.S. diet, ranking just below tuna In importance; (3) the mAjor portion of many of our most familiar types of sea food Is imported; and (4) freshwater species, led by trout, bass, and catfish, comprise about 9Z of our total fish diet. A compilation of PCB Data, representing the results of all the measurements known to NMFS of PCBf in fish used in the U.S. diet la presented In the Compendium of PCB data. It Includes data from government reports, and stats and private publications. An sffort vss made to Include current, unpublished data from marine laboratories. Tht compilation includes the results of a comprehensive literature search by the NOAA-Environaental Data Service (EDS). An EDS-ENDEX data search for unpublished date 146 HONS 05636? and a EDS-0AS1S search ware made, crossing marine food species with PCBs. Two significant sources of data--the 1975 FDA-Market Basket results, and the results of the extensive Fall 1975 New York State Department of Environment analyses were not available to NMFS at the time Compendium was published. However, FDA has Indicated that the 1975 Market Basket results do not differ dramatically from 1973/4 data. Preliminary results of the Hew York Study are Included in Table 13* Table 12 summarizes the PCB Information assembled In the Compendium relating to the 20 most Important kinds of dietary fish identified In Tablell. Although at one time or another, some PCB measurements have been made on many fish, Tablell points out the Inadequacy of information on PCB residues In the most important Items In the fish diet. Sampling and analysis have been sporadic and not designed to monitor treads In human exposure. Systematic surveys of neither the Important Items nor the species most likely to be contaminated have been undertaken. Very few recent (1975) measurements have bean reported. However fragmentary the data on PCB levels may ba, they do not Include a single measurement exceeding 1 ppm In any of the tan most important fish foods, except for Dover sole and crab taken In the Iroadlate vicinity of Los Angslss sewerage outfalls, and crab living on contaminated sediment* in Upper Chesapeake Bay, 147 HONS 056368 Rank// 1. 2. 3. 4. 5. 6. 7. 6. 9. 10. 11. 12. 13. 14. 15. 16. 17. 18. 19. 20. TABLE 12 Maximum Reported PCB Residue Levels In Fish Muscle (White Meat) Kind Range and Max. Level Reported (ppa) (see Compendium) Tuna Unclassified (mainly breaded) Shrimp Ocean Perch Flounder/Sole Clams Crabs/Lobster Salmon Oysters, Scallops Trout (excl. Lake Trout) Cod Bass Catfish Haddock Pollock Herring/Smelt/ Sardines (marine Pike Halibut Snapper Whiting 0-0.49 (1973) no data 0-0.167 (1971) 0-0.25 (1974) 0.6.3 (1972) 0-0.819 (1974) 0-4.9b (1972) 0-0.5 (1974) 0-0.43 (1974) 0-0.56 0-0.4 0.3-8.45 0-4.4d 0-1.16 0.0 (1972) (1974) (1974) (1974) (1974) (1974) 0* rt O O1 0-1.6 0-0.65 0-0.1 0.0 (1974) (1974) (1974) (1974) (1974) Mean Level : the U.S. Dl< Unknown* Unknown Unknown* Unknown* Unknown* Unknown* Unknown* Unknown* Unknown* Unknown* Unknown* Unknown* Unknown* Unknown* Unknown* Unknown* Unknown* Unknown* Unknown* Unknown* i These 20 Items compose 95% of the fish in the U.S. diet. * Probably well below 1 ppa aSingle Dover Sole taken near Los Angeles municipal sewerage outfall. The mean value of Sole samples taken In thla vicinity was 1.3 ppa in 1975. Mean values of other samples taken away from outfalls were less than 1 ppm In 1975. In any case, although Dover sole Is an Important food fish In norther California, It Is not taken commer cially in Southern California waters. ^Single yellovneck crab taken near Los Angeles outfall-mean of group Including this crab was lass than 1 ppm. A composite of Upper Chesapeake Bay crab contained 1.2 ppm In 1974. cTaken in 1974 from St. Croix River, Wisconsin--mean of 4 White Bass was 6.2 ppm. dTaken from Mississippi River, Minnesota--mean of a sample of 5 catfish was 2.5 ppm. H8 MONS 056369 No more than 1.6 ppm has been reported In Items 11 through 20 In importance, with the exception of freshwater baas and catfish living in river and lake areas known to be contaminated with PCBs. None theless, as emphasized in the Table, there have been no PCS measure ments on the important unclassified dietary fish items, and the analytical information is in no instance systematic enough to permit an estimate of the actual mean PCS values, hence human exposure. The strongest statement that can be made, based on the Incomplete data summarized in the Compendium, is that most of the important fish meat items eaten in the U.S. probably contain mean PCB residue levels well below 1 ppm. Fragmentary evidence also leads to the conclusion that the PCB levels in fish liver, roe and oil are approximately ten times that in the edible muscle and the PCB concentrations are greatest in the older larger oily fish, especially those that spend part or all of their lives in contaminated lakes, rivers or estuaries. T..bla 13 lists the location of the species where high PCB levels, exceeding the FDA 5 ppm guideline, have been reported. Although the trout, salmon and chub in Lakes Michigan and Ontario and tha Hudson River base, perch and eel constitute only e few percent at most of the national fish diet, in some ereee they are a significant nart of the fish diets. In order to discover what is presently known about U.S. flah consumption habits, a wide range of government and private sources 149 MONS 056370 TABLE U Areas Where at Least One Example of High* PCB Residue Levels Have Been Reported In the Listed Species Species Striped Bass Chub Carp Chain Pickerel, Alewife Coho Salmon Chinook Salmon Steelhead Trout Lake Trout Bass, Smalltnouth White Perch Alewife White Sucker Walleye Baas, Largemouth Yellow Perch American Eel, Crabs, Sturgeon Rock Bass, Catfish Bluefish Location Hudson River, New Jersey Lake Michigan Mississippi River, Minn., Lake Onandaga, NY Hudson River Lake Michigan, Lake Ontario, Hudson River Lake Michigan, Lake Ontario, Hudson River Lake Michigan, Lake Ontario Lake Michigan, Lake Ontario, Lake George, NY Lake Onandaga, Genessee River, NY Hudson River, Mohawk River, St. Lawrence River Lake Onandaga, Hudson River Hudson River Hudson River, Mohawk River Hudson River, Mohawk River, Black River, NY Hudson River, Mohawk River Hudson River Hudson River Hudson River Long Island Sound (muscle from one large oily fish contained 8 ppm) This Table includes recent data from New York State Department of Environmental Conservation not included in the NMFS Compilation. The Hudson River above Fort Edward, New York, is not contaminated with PCBs and the residue levels In fish are quite low. *Above the current 5 ppm FDA Guideline. i;>n HONS 056371 were consulted. A number of pre-1970 studies are available. Including a 1969 NMFS Survey of Fleh Purchaeea and a 1968-70 HFW Ten Slate Nutrition Survey. In addition, a few specialized reports have been published, Including annual FDA Regional Basket Surveys that do not include fish eaten in restaurants and Institutions or by sport fishermen, and surveys by the Sport Fishing Institute and a 1974 Texas A6M Analysis of Seafood Con sumption Patterns In Texas. However, the most up to date and comprehensive data ara available In the previously mentioned 1973 74 Seafood Consumption Study by the National Purchase Diary. Table 14 sunsaarlzes results of fish consumption demographic Information that defines the number of U.S. consumers who would exceed an arbitrarily set dietary micro-constituent level. To date, this kind of Information Is available only for total dietary fish and one subcomponent, tuna. Table 14 Indicates that 75 million !7.S. consumers sat more than the national average of 18.7 grams of fish per day and 29 million would exceed the present dietary lntaks limit of PCB (175mcg/day) If the 35 or more g/day of fish eaten was contaminated on average to 5 ppm, the current FDA guideline level. There Is, of course, no evidence thet the mean PCB level in fish la anywhere near 5 ppm. Nonetheless, should It In the future reach 1 ppm, then by an extrapolation from Table 14, an estimate can be made that about 200,000 U.S, consumers would be exceeding the 200mcg/day FDA limit. As 151 HONS 056372 TABLE 14 Number of U.S. Consumers Who Eat More Than X X (a/dav> 0 18.7 (national avg.) 35 50 100 150 Number (thousands of consumers) Total Fish Tuna 197,000 130,000 75,000 5,700 29.000 1,300 14,000 400 2,000 43 400 10 * Derived from graphs on pp, 21 and 23 of the Seafood Consumption Study, September 1973--August 1974 by the National Purchase Diary, Shaumburg, Illinois, 1975. Sponsored by the Tuna Research Foundation. 152 HONS 056373 another hypothetical caae, If the PCB level In tuna ehould ever reach 1 ppm, then about 5,000 tuna fleh consumers, those who eat In excess of 175 g/day, would exceed the 200ocg/day limit from tuna alone. Survey data do show that In general U.S. fish eaters Include a wide variety of fish Items In their diet. Thus, even If certain Items contain toxic micro-constituents at or near FDA guideline level, the great majority of consumers would receive only a small fraction of their average dietary Intake from these sources. Clearly, there Is a need for demographic Information, similar to that available on total fish and tuna, on the other principal foodflsh items In the U.S. diet; and a need to extend and better measure the hlgh-coneumptlon tell of the distributions, to get a correct picture of the number and type of consumers eating In excess of lOOg/day of any single fish product. IV. Conclusions At present, It Is difficult to estimate human exposure to PCB from eating fish, either for the population as a whole or for subgroups at higher risk. Fragmentary evidence suggests that the exposure to the population as a whole from PCB residues In Ingested fish -s probably well below 19mcg/day-consumar, based on PCB levels which are probably well below 1 ppm and 19g fiah consumed/day. 153 *0S 056379 Although It is important to know the total population exposure, the first priority should be given to defining the dietary intake of PCBs in special subpopulations at high risk by reason of high con sumption of fatty fish, or high sensitivity to the toxic action of PCBs. Such populations might Include pregnant women and young children, dieters, low income groups and sport fishermen living in coastal and lake regions where there is known PCB con tamination. Primate studies suggest that pregnant women and young children will be particularly sensitive to PCBs, and dieters, and in some areas low income groups may rely heavily on fish for food. For each subpopulation, the amount and kind of fish In the diet and the PCB residue levels in the fish eaten must be obtained in order to determine exposure. None of the necessary Information to define the human PCB exposure in subpopulations Is presently known. The first priority in systematically monitoring and analyzing for PCB residues in fish should therefore be among the oily fish such as salmon, trout, mackerel, shad, sardines, herring, bluefln tuna, sable fish, white fish, striped base, and blueflsh, that have been identified as significant food items In the diets of special subpopulations of consumers. Although onl, minimal dietary exposure to PCBs may occur through food, there is evidence of potentially significant exposure 154 HONS 056375 in those sub-groups of the population who regularly consume fresh water fish from waters which are contaminated with PCBs at lsvels which exceed the FDA temporary tolerances. These sub-groups of the population would Include sports fishermen and others who consume locally caught freshwater fish. In surveys conducted by the State of Mew York and FDA, samples of 17 species of fish were collected from 10 points on the Hudson River between Glens Falls and Alpine, 13 samples were collected at Glens Falls and 68 samples below Glens Falls. The data from these surveys indicate that 53 out of 68 samples of fish collected at or below Fort Edward (approximately 7 miles below Clans Falls) con tained PCBe in excess of 5 ppm. The average level of PCD contamina tion in these 68 samples was 31.3 ppm. In contrast, only trace levels were found in the 13 samples obtained at Glens Falls. A high proportion of flnflsh and eels from all sanpllng points below Glens Falls contained PCB levels in excess of 5 ppm. The Fort Edward sampling point had the highest levels for the special examined, with individual samples ranging from 20.1 to 178 ppm. Twenty-sight of the 33 samples of coho and Chinook salmon from Lake Ontario exceeded 5 ppm, with a range from 21 ppm to 26.6 ppm. Of the 6 samples of lake trout tested, results were in ths range of 10 to 15 pi ... All other species of fish sampled from Lake Ontario were at 5 ppm or less PCBs. The Great Lakes represent another area of the nation contamina ted with PCBe. The Michigan Water Resources Commission has reported 155 HONS 056376 that many surface water samples contained PCBs at concentrations above the detection limit of 10 ppt. (10 significant levels of PCBs have been found in rivers and streams discharging into the Creat Lakes. Effluents from wastewater treatment plants servicing industrialized communities have been found to be highly contaminated. Sampling surveys of Great Lakes fish have shown that most species tested contained detectable levels of PCBs and that residue levels were generally proportional to fish size (age) and highest in the predator species. Except for whitefish, the species of commercial or sport interest (trouts and salmons) from Lake Michigan were found to be highly contaminated with PCBs. Data obtained from lake trout collected from various areas of Lake Michigan show mean PCB levels ranging from 3.06 ppm to 11.93 ppm. The Michigan Department of Public Health has recently completed a study (Humphrey et al., 1976) which attempted to assess some of the consequences of human exposure to PCBb from the consumption of sportsfiBh caught in different areas of Lake Michigan. The study included-exposed and control subjects from five areas of Michigan bordering on Lake Michigan. Exposed study subjects were those individuals who consumed at least 24 to 26 pounds of Great Lakes fish per year. Control subjects were those individuals who consumed less than six pour of Creat Lakes fish par year. A preliminary assessment of t.ie findings in the study indicate that the moat frequently recorded quantity of fish consumed by the flONS 056377 study participants was In the 24-25 pounds per year range. The highest recorded fish consumption over the two year period of the study was 180 pounds per year and the highest single season consumption was 260 pounds. Mean PCB levels In whole Lake Trout are reported as 18.93 ppm in 1973 and 22.91 ppm in 1974, and 12.17 ppm in Coho Salmon in 1973 and 10.45 ppm In 1974. However, comparisons of PCB levels In raw vs. cooked fish Indicate that actual human exposure to PCB from fish consumption Is less than might be expected from the raw fish dats. This is not unexpected since preparation (trimming away fatty tissue) and cooking have been shown to decrease the concentration of PCBe. For example, the PCB level in cooked Lake Trout consumed by the study participants ranged from 1.03 ppm to 4.67 ppm; In cooked salmon from 0.48 ppm to 5.38 ppm; and In other cooked fish from 0.36 ppm to 2.06 ppm. These levels are decidedly lower than the level of PCB contamination reported in raw trout and salmon. PCBs vara found In 501 blood and breast milk specimens col* lectsd from study participants during the etudy. The values ranged from a low of 0.007 ppm in blood in the control group to e high of 0.366 ppm In the expoaed group. Although there was a vide range of blood values for each quantity of fleh consumed, thera was a highlv elgnlf.' ant correlation between the reported quantity of Lake Michigan fish consumed and the concentration of PCB in the blood of study participants. Using a natural log transformation 157 HONS 056378 of PCB values, the correlation between amount of fish consumed and PCB blood levels was significant (t*6.24, p 0.0001), with higher reported fish consumption being associated with higher blood PCB levels. No annual variation in PCB blood levels in humans could be demonstrated. The mean PCB blood values for the control and exposed groups did not appear to change markedly from 1973 to 1974. In addition, abstinence from Lake Michigan fish consumption for a period of 90 days or more did not change the PCB blood levels sig nificantly. PCB blood levels over the abstaining period shows variation but no steady decline in PCB. In fact, more subjects showod no change or a rise than showed a decline in PCB blood levels over time. The calculated quantity of PCB ingested by eating Lake Michigan fish averaged 46.5 mg/year and ranged from 14.17 to 114.31 mg/year. PCB Ingestion for each Individual was determined by proportioning the reported annual fish consumption by frequency of species eaten and the cooked fish PCB levels for those fish. The community average for cooked fish was used in Instances were cooked fish determination were not available for a study participant. Because fish consumption was found to vary from year to year the average annual consumptit for each individual for the two baseline years of study was used in each case. I 5H M0NS 056379 Results from this study Indicate the strong likelihood that the maximum allowable Ingestion of 1 meg/kg body veight/day recommended for protracted exposure to FCB Is being exceeded by the majority of the exposed group participants, and by Inference, by a substan tial number of Individuals participating In sport fishing and who consume quantities of contaminated fish. The calculated mean dally dose received by the exposed group la 1.7 meg/kg/day and ranges from 0.49 meg to 3.94 meg/kg/day. If the average annual rate of FCB Ingestion from fish indicated by these study results ware con tinued over the years, and if net accumulation occurs, the average sports fisherman consuming contaminated fish could receive a total FCB dose equal to the 200 mg safety limit in approximately 4.3 years. Under the same set of assumptions, individuals consuming greatet than average amounts of contaminated fish would reach tha total dose level sooner. No adverse health effects or group of symptoms could be iden tified In the exposed group that were clearly related to FCB exposure. This implies that exposure to FCBs from eating contamina ted fish at the levels observed and the presence of FCBs In these exposed persons has not caused any observable adverse health effects at the time of the study. However this does not exclude . ths possibility that affects too subtle for detection are occurring or ths possibility of long-tsrm adverse health effects. 159 HONS 056380 Considerable scientific Jnterosc lias centered on the Yusho incident in Japan where in 1968 human intoxication with Kanechlor 400, a PCB manufactured In Japan, was noted when a heat exchange leaked this PCB into rice oil (''Yusho" oil) which was consumed by Japanese families. The typical clinical findings included chloracne and increased pigmentation of the skin, increased eye discharge, transient visual disturbances, feeling of weakness, numbness in limbs, headaches and disturbances in liver function. Most of the babies born to mothers with Yusho had skin discoloration which slowly regressed as the children grew in size. Adult "Yusho" patients had protracted clini cal disease with a slow regression of symptoms and signs, suggesting a slow metabolism and excretion of the PCB in humans, probably resulting from a long biological half-life. A review of the literature extent in 1972 led to the following facts with respect to this tragic Incident in Japan: 1. The average PCB content of the rice oil In the doseresponse epidemiologic study was 2,500 ppm. 2. In this dose-response epidemiologic study, the average cumulative lntaie of PCBs, leading to overt symptomatology, was 2000 -g. In this same study, the lowest dose leading to overt symptomatology was 500 mg. 160 M0NS 056381 3. The toxicity seen was the result of the Ingestion of the PCBt contaminant in the rice oil. A aunsnary updating of the "Yusho" incident was presented by Professor Kuratsuna at the National Conference on Polychlorinated Biphenyls, Chicago, Illinois, November 19, 1975 (Kuratsuna et al., 1976). This report affects the original conclusions enumerated above. 1. PCBs Content of MYusho" Oil. The flret reported PCB determination wee made on a canned contaminated rice oil produced on February 5, 1968, and whose consumption was associated with overt ayvptomology. One group of Yusho patients was associated with ingestion of this contaminated canned rice oil produced or shipped on February 5 and 6, 1968 (Kuratsune at al., 1972). The detailed epidemiologic atudlee are given in the papers by Kuratsune et al., (1969) and Yoshlmura (1971) and sum marized in English by Kuratsune (1972), Kuratsune et el., (1971) and Kuratsune et el., (1972). Of 325 petlente queried, 166 of 170 ussd canned rice oil produced or shipped on February 5 and 6, 1968, and 143 of 155 used bottled rice oil shipped from February 5 to 15, 1968. Tsukamoto et al., (1969) determined that the organic chlorine * ntent of canned rice oils produced or shipped on February 5, 1968, was 1000 to 1500 ppm, by chemical and activation analysis. Since Kanechlor 400 had a 48 par 161 HONS 056382 cent chlorine content , ione a 11 r t hu t ed aM ttu <> ry.m i. chlorine to this specific eh 1 or i n.it Oil Mplicnv I , i ti,-n Mil-. canned rice oil contained, jj> an average, db()0 ppm iyMh. Kuratsune et al., (1969) reported that a qualitative GLC survey of 109 samples of bottled rice oil shipped between October, 196/, and October, 1968, showed "significant" PCBs content only in those samples between February 7 to 10, 1968. The largest amount was reported in the sample of February 7, 1968, but the quantity was not given. No bottled rice oil samples were available for February 5 and 6, 1968. However, in the paper by Kuratsune et al., (1971), the most marked contamination of bottied rice oil was for that shipped on February 7, 1968; and the maximum chlorine content was 462 ppm. One may assume that this number applies to the sample of bottled rice oil of February 7, 1968, giving this sample a PCBs content of 924 ppm. Kuratsune et al.t (1976), reporting the work of Nagayama et al., (1975b), gives the PCBs analyses for three samples of toxic "Yusho" oil, used by three independent families with "Yusho", as approximately 800 to 1000 ppm. These were GLC/MS measurements. Nagayama, et al,, (1975b), described these samples as follows; "Rice oil used hy patients with Yuso ("Yusho oil"); 3 samples of rice oil used by 3 independent families with Yusho in 1968. These samples had been kept in glass bottles with glass stoppers at room temperature In our laboratory until 1974 when the analysis was made." I 62 HONS 056363 Kuratsune (1976) pointed out that thaae thraa samples ware canned oil and considered to have been produced and shipped on February S or 6, 1968. This Is also suggested In footnote a, Table 5, In the paper by Kuratsune et al., (1976). It waa reported prevloualy that, baaed on organic chlorlna determinations, Tsukaaoto et al., (1969) reported an average PCS content of the canned oil of February 5, 1968, to be 2500 ppm. Now, based on BLC/MS, we have a PCB content of ca 1000 ppm for canned oil sample of February 5 or 6, 1968. What ara the posalble explanations for this discrepancy In values? One possibility Is beaed on the aesuaptlons essoclated with the method of measurement by Taukamoto et al,, (1969); namely, determination of total organic chlorine. Thus, any orgnlc chlorine preeent In the rice oil, which was not a chlorinated biphenyl, would be Included in the FOB analysis. Some possibilities include: chlorinated dlbenxofurans, chlorinated naphthalenes, chlorlnatad paraffins, other chlorinated allphatlcs and non-blphenyl aromatics. Another possibility la related to the distribution, with tlaw, of th-v contaminated rice oil samples. February 5, 1968, ccurred on a Monday. Tha first rice oil samples reported to contain ?CBe were produced on thla date (Taukamoto et al.; 1969). If tha laak first occurred sometime over the weekend (February 3 and 4, 1968), and 163 MONS 056384 production was resumed Monday morning, February 5, 1968, then one would expect the samples of this date to be higher in PCB content, Therefore, one would expect the first batches Monday morning to be highest with a dimunution of PCB content with time. Tsukamoto e_t a_l., (1969) reported the organic chlorine content of six canned oils produced Monday, February 5, 1968, as follows; 1020, 1170, 1070, 1080, 1030, and 1500 ppm. In the survey of bottled oils, Kuratsune et al., (1969) reported that for those samples where PCBs were found, the greatest amount was in the samples of February 7 (Wednesday); and after February 11. It was Inferred from the paper by Kuratsune et al., (1971) that the PCB content of the bottled rice oil samples of February 7, 1968, was 924 ppm. In addition, in Table 5, Kuratsune et al, (1976) gave a value of 134 ppm PCB for a rice oil produced on February 10, 1966. Thus, the available data does confirm a diminution of PCB contamination of the rice oil with time. This Is summarized below: Pate Source Feb. 5, 1968 canned Feb. 5 or 6, 1968 canned (?) Feb. 7, 1968 bottled , Feb. 10, 1968 bottled After Feb. 11 1968 bottled PCB content (ppm) Method 2000-3000 organic Cl ca 1000 GLC/MS 924 organic Cl 134 7 just detectable CLC (?) 164 HONS 056385 The resolution of the varying PCB values for the canned oil cannot be made. However, Kuratsune (1976) confirmed that the production of rice oil had been suspended for an unknown period of time prior to February 6, 1968, when It resumed. 2. Dose-Response There were 1291 "Yuaho" patients as of April 30, 1975 Oeme, 1975). Of this totAl group, the dose-response relationship was developed from a detailed analysis of 146 patients (Yoshltoura 1971; Kuratsune et el., 1972). These 146 patients all consumed contaminated canned rice oil produced on February 5, 1968 contained 2000 to 3000 ppm PCBe as Kanechlor 400 (Tsukaaota et al., (1972) estimated that the lowest dose producing symptoms was 0.5 ga. If the PCB content of this canned oil was ca 1000 ppm, as discussed In the previous section, then the average dose producing an overt effect was 0.8 gm. and the lowest dose producing an overt effect was 0.2 gm. 3. Causative Factors. Originally, rice oil contaminated with a heat exchanger, Kanechlor 400, a polychlorinated biphenyl, was associa ted with "Yuaho" symptomatology (see discussion In section 165 HONS 056386 1 above). PCBs were Identified in the contaminated rice oil consumed and in the blood and tissues of Yusho patients. Therefore, the effects seen were attributed to PCBs. In the review by Kuratsune et al. (1976), a new factor was introduced into the system; namely, the canned rice oil was also contaminated with chlorinated dlbenzofurans to the extent of S ppm. In addition, in this same paper by Kuratsune et al., (1976), they presented data of Bagayaxna et al., showing polychlorinated dlbenzofurans In the liver and adipose tissue of Yusho patients, while none was found in that of a control group. The ratio of PCBs to PCDFs in Kanechlor AGO, a Yusho oil of February 5 or 6, 1968, in adipose tissue and liver from a Yusho patient were 50,000; 200; 144; and 4 respectively. Thus, relative to Yusho oil, the liver with a PCB/PCDF ratio of 4 to 1, appears to selectively concentrate PCDFs relative to PCBs. Relative to PCBs, PCDFs are about 250 times higher In the rice oil than in Kanechlor 400 (Nagay~ma et al, 1975b; Kuratsure et al., 1976).* They also stated that PCDFs ov / be a factor in Yusho disease. In another should be noted that ve are comparing chlorinated dlbenzofuran content of an unused Kanechlor 400 to that after prolonged use in a heat exchanger. 166 HONS 056387 paper Nagayama ec al., (1975a) reported that it "la said that the toxicity of PCDF la said to b fro* 200 to 500 times that of PCB". At tha lower factor (200), con taminated rice oil would be expected to be twice as toxic ea K&nechlor 400, Also, in this same paper they stated that the symptoms seen in Yusho patients seemed to be more severe then would be expected Just from the PCB intake associated with the oil. If PCDF is 200 to 500 times mors toxic then PCB (Nageyama at al., 1975a), than contaminated rice oil would be 2 to 3*5 times more toxic then that expected from its PCB coo* tent, elone. Kuratsune at el., (1972), estimated that if the average amount of contaminated rice oil ingested by e "Yusho" exposed person, is related solely In terms of Its PCB equivalent, then the amount of oil neadad to be Ingested, on that basis, would be 1600 ml to 2800 ml. At s PCB content of 2500 ppm, this is an Ingestion of 4 to 7 gn of PCBs equivalent; whereas at a PCB content of 1000 ppm, this Is an Ingestion of 1.6 to 2.8 gm of PCBs equivalent. Conclusion The complexities end uncertainties associated with the most recant reports of the "Yusho" incident in Japan, make it extremely difficult to quantify possible human health effects resulting from exposure to PCBe elone. 167 HONS 056388 V. Occupational Exposure to PCBs The earl tost reports of adverse health effects due to exposure of workers to PCBs In this country arc probably those of Schwartz (1936) who described skin lesions and symptoms of systemic poisoning among workers who were said to have inhaled chloro diphenyls; their complaints included digestive disturbances, burning of the eyes, impotence, and hematuria. Patch tests with the chlorodiphenyls were negative, and Schwartz speculated that mechanical plugging of the follicles of the skin as the fumes solidified on it were respon sible for the skin lesions; the chlorine present in the products were thought to then exert an irritating effect on the plugged fol licles, and to thus cause suppuration. No quantitative data were reported, but a number of preventive practices were recommended. There have been numerous reports over the ensuing years (Schwartz and Bartow, 1942), (Greenburg et al., 1939), (Drinkes et al., 1937), (Good et al., 1943), (Schwartz, 1943) and (Meigs, et al. 1934) of cutaneous eruptions and of systemic manifesta tions (sometimes fatal) as well, among marine electricians, machinists, capacitor and transformer manufacturing workers, and others occupationally exposed to PCBs. However, In many of these reports the exposures arc described as having been to mixtures of chlorinated hy^ ocarbons, quite often of chlorinated naphthalenes and PCBs. 1 f>R MONS 056389 The skla lesions described by Schwartz (1936) have come to be deslgneted generally as "chloracne." Chloracne can be produced by a number of chemical compounds, including chlorinated dlbenzofurans and certain Isomers of the chlorinated dlbenzodloxlns (Kimbrough, 1972). Oily skin and large pores seem to predispose to the disease, while the opposite is the case for smooth, tender skin (Kimbrough, 1974). Chloracne also has occurred among workers engaged In the production of 2,4,5-T (Poland et al., 1971). Part of the chloracne lesion resembles adolescent acne, but It is generally more severe, and lesion distribution Is inconsis tent with, although it may be superimposed upon, adolescent acne. It la known that chloracne can be produced by either the systemic absorption of chlorlnatsd biphenyls or ths direct application of chloracnegenlc compounds to tha skin. Systemic effects sometime* result after occupational chloracne has manifested Itself; these may Include loss of appetite, nausea, edema of the face and hands, abdominal pain, vomiting, and burning and eoreness of the eyes. No fully satisfactory explanations have been made of the develop ment of chloracne. Chloreene Is generally very persistent, end there is the preferred control measure. An excellent review of this subject is that by Crow (1970). The U.S. /Ccupstlonel standards for PCBs are; 8-hour timeweighted average exposure limits of 1 mg/iP (skin) for the 42X chlorinated product, and 0.5 mg/m^ (akin) for tha SAX chlorinated product (CFR, Title 29, Part 1910.93). The National Institute for Occupational Safety and Health hee under way the preparation of L69 HONS 056390 criteria for a recommended standard for occupational exposure to polychlorinated biphenyls. In addition to an environmental exposure limit, this document will provide comprehensive recommendations for medical surveillance, safe work practices, and engineering controls to ensure employee protection during occupational expo sure over a working lifetime; it is scheduled for transmittal to the Department of Labor in late 1976 or early 1977. NIOSH also i9 undertaking certain studies of employee populations (mainly in U.S. capacitor factories) in order to better define the nature and extent of any chronic and/or life-shortening effects of exposure to PCB. Practically all of the currently available information on worker health and PCB exposure is found in foreign literature. For example, Karppahen and Kolho (1974) reported on relationships between the concentration of PCB*s in the blood of all, and in the adipose tissues of some, of 29 persons in "good health"* from *Kolho has stated that the capacitor plant workers received quar terly health examinations, and, in addition to the clinical examina tion made at the time of the Investigation, serum alkaline phos phatase, GOT, and GPT activities were determined; the 6 employees with the highest blood concentrations of PCBs also had BSP excre tion tests performed. All results were normal. In view of claims based on animal experiments that PCBs can induce liver microsomal enxyme activity, the authors also determined the half-time of ant'^yrin before and after phenobarbital induction (1 mg phenobarbital/kg tody weight/day/3 days) for 6 capacitor plant employees and 6 controls; no epzyme induction was observed. Further, because of claims that ICB's have effects on steroid metabolism, 4 capacitor plant workers were tested for ACTH (serum, presumably); all results were reported to be within the normal range (letter from S. Hemberg to A. C. Kolbye, Jr., 12/23/75. 170 HONS 056391 V three different employe groups Id Flnlend. None cf the employees Had no hlecory of occupational exposure to PCBe, six of them had handled PCB samples in an analytical laboratory, and the other eleven of them had been employed for 6 years in a capacitor fac tory where Aroclor 1242 was used as the impregnating fluid. It was stated that average PCB concentrations in the air of the capa citor factory had not exceeded "Internationally accepted limits**," and that special attention had been given to skin protection. Table IS shows the observed tissue concentratins of PCB's; the authors were unable to detect any biological effects of the approxi mately 50-fold larger PCB concentrations in the blood of the capa citor plant employees, compared to the "unexposed" control group. Ouv et al., (1974) conducted a survey to determine the degree of absorption and the health effects of exposure to "electrical grade" Aroclor 1242 (that "did not contain any lmpurltlea") for varying periods of time. PCB concentrations in the air of a capa citor factory in New South Hales, Australia*, were measured, and 34 occupationally axposed employees (15 sales, 19 females, ranging in age from 33 to 55 years) were examined and compared with volun teer controls (23 sales, 7 females ranging in age from 20 to 50 *Tha Joint 11/ 'WHO Committee on Occupational Health, in its sixth report (World Health Organization Technical Report Series No. 415, 1969), recommended for international adoption a "safe concentretlon zona" of 1 mg/a3 for chlorinated derivatives of diphenyl. *Aroclor 1242 is not manufactured in Australia (Ouv at al., 1974). 171 HONS 056392 TABLE IS Concentration of PCB's In blood Fat basis mg/.fcg Average Range In adipose tissje Fat basis ">R/kR Sample no Workers in capacitor factory 313 100-700 l 200 2 11 3 160 4 265 5 635 Persons handling PCB's in analy tical laboratory Persons without any special exposure to PCB's 53 5.4 33-71 3.6-9.9 not analyzed 1 2.3 2 1.5 From Karppanen and Kolho (1974) 172 HONS 056393 year*) having no history of occupational exposure to PCB'a. Study parameters Included occupational and medical histories, PCB-lnblood concentration estimates, and liver function (serum bilirubin, alkaline phosphatase, total protein, and GPT, and BSP excretion) testa. The authors noted that exposed workers tended to complain of eye, face, and akin "burning," that the PCB '"fume' ... has a pungent small which often causes persistent body odour," and that the' employees with higher blood concentrations of PCBs complained most often of the skin lesions (one case of chloracne, flva cases of "an eczematous rash on" the legs and hands), although there apparently was poor correlation of blood PCB's with the severity of the complaints. No significant health affects ware observed among those of the 34 workers whose blood PCB concentrations wars below 200 ppb. There was a statistically significant difference betveen the blood PCB concentrations of tha exposed group and the control group (P lets than 0.01). Table 16 shows tha concentrations of Aroclor 1242 In tha capacitor plant air prior to and after exhaust ventila tion system "Improvements" bed been made. (The Australian National Health and Medical Research Council recommended (Atmospheric Contaminants, 1970) axpoaure limit values of 1.00 mg/m^ for PCBs of 42X and 54X chlorine content, respectively). Table 17 shows that there was no lowering of blood PCB concentration among those workers tested two months after tha Installation of a more 173 HONS 056394 i'ABLF. J 6 Arocloro 1242 concentrations in the air inside capacitor plant before and after improvement of exhaust ventilation system Aroclor concentration No. Areas in the Impregnation Room in tng/m^ Before Af ter 1 Area in the unloading tank in front of exhaust register from operator's breathing zone 1.44 0.75 2 Area in the unloading tank not in front of exhaust register 2.22 0.7 3 Ceneral atmosphere near tank 1.08 0.18 4 Soldering area 0.32 0.08 From Ouw et al (1974) 174 HONS 056395 Mean blood Aroclor 1242 levels before and after improvement of exhaust ventilation and the recommen dation to use "suitable impervious" gloves. Group Before After Statistical differences Mean blood Aroclor levels In ppb. Retention times relative to Aldrln 1. 0.69 1.31 1.41 281.6 135.1 58.41 477.2 225.4 524.7 Not significant difference (P 0.01) Not significant difference O 0.01) Significant difference (P 0.05) From Ouv et al (1974) 175 HONS 056396 efficient exhaust ventilation system and the concomitant recom mendation to wear "suitable impervious gloves" in order to reduce PCB absorption through the skin, The authors suggested that a failure to adhere strictly to the glove recommendation might expla.n the continued elevation blood PCB levels. Another possibility might be that PCBs were continually mobilized from storage in adipose tissues* The Japanese Ministry of Labor (Regulation for the Prevention of Disturbances due to Specified Chemical Substances, under the Act for Safety and Health of Workers, of April 28, 1971) established an occupational environmental exposure limit for PCBs of 0.5 mg/ro^ at 25 C, one atm (Ad Hoc Committee, 1974). Japanese Importation of PCBs commenced around 1950, and early uses were as dielectrics in capacitors and transformers. In 1954, Kanegafuchi Chemical Industry Co., Ltd. started PCB production in Japan and It was at about this time that chloracne eruptions among workers were first reported. Slightly earlier (1953) incidents, however, were reported by Kara (1969), of changes attributable to PCBe. in blood and urine findings among capacitor factory workers. , According to an Ad Hoc Committee sponsored by the Japanese Environmental Agency (1974), PCB concentrations In the air of a Japanese :apacltoi factory (where the ma^or PCB constituent was "diphenyl pcntachloride") were found to range from 0.37 to 6.75 mg/m^; this was between the years 1953 and 1957, and prior to the 17b ttONS 05639? date when the aforementioned occupational exposure limit ve* promul gated. The highest concentration ves measured where PCBs were heated to drive out entrained/dissolved air. Admittedly, the analytical methodology for PCBe In those years was not as refined as that now In use, but It la thought likely (authors) that soma employees were exposed continually at concentratlona of several milligrams par cubic meter. The Janapese literature Is said to have contained no further reports on PCB-ln-air measurements until the spring of 1972, Just prior to the suspension of their usage. The production of PCBs was discontinued In Japan in June 1972, and their Importation was discontinued In the following September. At that time Rasegawa et al., (1973) reported PCB measurements In the air of one PCB (Kanechlors 200-600) production plant (0.005-0.02 ppm), four capacitor manufacturing plants (Kanechlor 300 used) (0.01-0.05 ppm), and one plant where PCBs (Kanechlor 300) were used as a heat exchange medium (0.002 ppm). Peak values of 0.17 ppm and 0.67 ppm, respectively, were found In a capacitor plant "alr-rlddance" process area and In a capacitor Impregnating area whera tank leak age had occurred. Estimated (preausiably 8-hour workday) timeweighted average exposures of affected employees vers 0.02-0.03 ppm. Airborne PCBs in the capacitor plants were reported to con sist of 70-801 vapor for material corresponding to Kanechlor 200, end of particulates larger the.. 0.1 micron for Kanechlor 300. The authors attributed the 3/1 vapor/partlculate mix to selective evaporation of low Bolling components of Kanschlor 300. 177 HONS 056398 Although no tliedaUlriitHiuLs wci'B available, llac^dWd el dl. , (1973) estimated that environmental conditions in carbonless copy paper manufacturing facilities (using Kanechlor 300) were roughly similar to those found in capacitor factories. Beginning in February 1971, PCBs were phased out as the microcap9ular solvent in this process, being replaced by "SAS" or KMC-oil;" however, during the period from November 1972 to January 1973, when occupa tional health surveys were being performed relative to SAS or KMCoil toxicity, PCB concentrations of 0.013-0.4 ppb were measured in the environments of these facilities. Ceneral environmental con centrations of PCBs at that time were less than 0.0005 ppb, and outdoor PCB concentrations around the facilities were 0.0043 ppb (Hasegawa et al., 1973) and 0.009 ppb (Sato and Hasegava, 1974). In an office room where carbonless copy paper was used, an air concentration 1.1 meg PCB/nr* was measured, and in the carbonless copy paper storage area of a post office, PCB concentrations of 8.7-21.1ccg/m^ were detected (Nishiyama et al. , 1973). The Japanese Ministry, in its "Regulations for Physical Examinations for the Prevention of Disturbances due to Specific Chemical Substances," promulgated on October 14, 1971, advised PCB users to examine employees for dermal andhepatic "symptoms" and their anamnesis, "subjective symptoms" such as anorexia and asthenia, and urine urobilinogen, in the first (pre-employment?) physical examination, and to "sut/ey" working conditions, blood tests and liver function testa in the "secondary" (periodic re-?) 178 Hons 056399 examination. A 1973 notice adlvsed those users of PCB's as hest exchange fluids to perform comprehensive health surveillance (Environmental Agency of Japan 1974). Yamamoto (1974) reported "positive" findings In 37 employees of 323 (total employment 706 In 51 establishments) who were given "special" physical examina tions in 1971. Hara (1969) reported a 20-30Z incidence of dermal "symptoms," consisting of "distinctive hair follicles" in exposed areas such as the face, neck, and forearm, and pimple-like skin eruptions of the face and neck, among employees In a capacitor factory that had been In operation between 1953 and 1963 (pantachloroblphenyls) 1953-1957; Kenechlor 300, 1958-onward). No other, e.g., hepatic, dysfunction was observed, and no quantitative information was reported In the reference from which the preceding report was obtained; however, in a follow-up survey of workers in capecltor factories, Bars st al (1974) reported that by one year after the suspension of PCB usage skin findings had become milder. Hasegawe et al., (1973) reported on a 1972 survey of capacitor factorise, in which they noted that pereons working in environments that contained 0.2-0.3 mg PCB's/tr* (0.02-0.03 ppm) shoved dermatolpgic ailments that Included "brown chromodermatosls" of the dorsal Joints of the hands . d fingers and nail bed, and "acneform exanthema." Sevsral cases of comedo or acneform exanthema of the jaw, back, and thighs were seen also. These signs were no longer observed one month after the cessation of the handling of PCB's. 179 HONS 056400 In a factory where Kanechlor 300 was employed as a heat exchange medium, the environmental concentration of PCB's was reported to bo 0.02/mj/m^, and no dermal ntani ft'Ktat ions were observed among Its employees (Hasegawa et al., 1973). Nor did Hashimoto (1974) observe any abnormalities among 236 workers in such a facility, to whom he administered ''examination centered around liver function tests." In 1972, when Hasegawa et al., (1973) performed a health sur vey of workers in carbonless copy paper factories, i.e., 2 years after the use of PCBs In such processes had ceased, they noted no dermal effects, and no liver function, blood, or urine test abnormalities were seen, with the exception of "slight abnormalities in lipid metabolism". PCB levels in the blood of these workers were reported as "approximately 0.01-0.02 ppm," or what amounted to a decrease to 10% of the levels during the period of PCB usage. The authors' data Indicated that PCB's collected from the work atmosphere had apparently degraded to a product containing one less chlorine atom than the average number of chlorine atoms in the PCB's handled in the factory, and that, conversely, the PCB's found In the workers' blood contained one more chlorine atom than did the PCB's handled in the factory, i.e., if the PCB's found in the workers' blood were Inhaled at the workplace, it can be surmised that the di- and trlchloro- biphenyls disappeared rapidly from the body, whereas the tetrachlorobiphenyl was metabolized slowly. I MU M0NS 056401 Haaegava et al., (1973) concluded that blood analysis Is a feasible method for determining body burdens of PCBs, and "pro bably mors useful than the technique presently used by extracting adipose tissue and using it for PCB;" urinalysis for "PCBs did not appeal to them aa a viabla monitoring method. They noted, how ever, that there wae no correlation between length of employment (and presumably, duration of ezpoeure) and the amount of PCBa accumulated in the blood, i.e., with continued exposure, PCB level# in the blood did not Increase linearly; in light of the demonstrated slow excretion rate of PCBs from the body, the authors conjectured that some different blotransformatlve mechanism comae into play after blood PCB levels exceed approximately 1 ppm (perhaps fat storage). This would stem to belle their confidence In blood analysis ae a reliable index of exposure. Kitamura et al., (1973) reported that the mean PCB level In the blood of 10 eepecltor factory workera was 0,62 ppm (0.32* 2.1 ppm) Immediately after the cessation of PCB usage, and 0.31 ppm 3 months later, when almost all of the subject were observed to have akin signs. It seems significant that the blood PCB con-* centratlon* observed In thla study vsre several fold hlghar than those seen by Haeegewa et el., (1973). Kitamura et al., (1973) attributed the skin effects to PCB'e accumulated In the workers* bodies during the period of PCB usage. No other abnormalities were reported. 181 HONS 056402 Sagaml et al., (1973) reported the case of a housewife in whom they observed skin effects several years after she had left 6 years of employment In a capacitor factory. She was found to have PCB concentrations of up to 0.13 ppm in her blood and of 42 ppm in samples her subcutaneous adipose tissue; this contrasted sharply with the findings of Hara et al (1974) that the blood PCB levels of 3 workers who remained at the same factory had decreased to 0.03 ppm and below, one year after the cessation of PCB usage (from levels of 0.03-0.3 ppm during the time of PCB usage (Hasegawa et al., 1973). N.B. Some capacitor manufacturers In the U.S. use epoxide-type alkylating agents as additives to prolong the service life of PCBs. EPA sec. 308 responses from the General Electric Co.f probably contain identifying data. VI. Air and Water Exposure to PCBs As indicated previously, nominal human exposure to PCB'e in die U.S. population may occur from air and water. Samples of ambient air were collected in suburban areas of Miami, Florida; Jackson, Mississippi; and Fort Collins, Colorado. Preliminary results (Kutz and Yang, 1975) for samples taken in April, May and June of 1973 show that PCBs were present at all locations. Although the data varied, the average concentrations at each of the three locations was approximately 1Q0 nanograms per cubic meter. Initial 182 MGNS 056403 identification of the PCBs. indicated that they were Boat conparable to the Aroclor 1254 standard. Dennis (1975) has reported that data gathered froa monitoring activities of surface weters and bottoa sediments of the major drainage basins of the United States indicate the widespread occurrence of PCBs in both surface water and bottom deposits. A preliminary assessment of PCB levels shows mean residue levels in water ranged from 0.01 to 0.05 mcg/llter. The 0.05 mcg/llter were found in the South Atlantic Slope and Eastern Gulf of Mexico drainage basin. In general, the loweat PCB residue levels were found in drainage basins west of the Mississippi. Klelnert (1975) has summarized the work completed by the Wisconsin Department of Natural Resources to identify some PCB sources in the environment in Wisconsin. Effluents froa cooling watsr in aluminum foundries contained PCBs renglng froa 11.5 to 335 ppb. Investigations revsaled the common source to be leaking hydraulic fluids containing PCBs which were ussd in die cast machines. Effluents from paper Bills that rscycle vastapapara has maaaureable discharges ranging from 0.01 to 25 ppb. Snow saaplss (Klelnert, 1975) were collected early in 1975. Analysis of the snow melt vater from Racine, Kenosha, Madison, end Milwaukee revealed concentrations froa 0.17 to 0,24 ppb. The author concludes that these values suggest that fallout of PCBs froa the air may be a principal source of PCBs entsrlng the waters 183 HONS 056404 of the. Statu. Hesse (1975) Indicates that similar to the studies in Wisconsin* not all industrial affluents tested in Michigan contain measureable PCE levels. In testing over 900 industrial samples, approximately 40 percent contained PCBs above the 0.1 mcg/liter laboratory sensitivity limit. Twenty-three percent of the indus tries tested had greater than 0.5 mcg/liter, 18 percent greater than 1 mcg/liter, 6 percent greater than 10 mcg/liter and 2 percent greater than 100 mcg/liter. Although much of the PCBs entering municipal waste treatment facilities are removed and become Incorporated into the waste sludge, a sampling of 58 municipal wastewater treatment plant affluents throughout Michigan in 1973 showed an average concentra tion of 0.52 mcg/liter. Concentrations of PCBs are much higher in the sludges. The average for ell plants was 15.6 mg/kg with individual values as high as 350 mg/kg. Since sewage sludges are conanonly disposed of by incineration, spreading on agricultural land, or placing in landfills, the addition of PCBs to the environ ment is obvious. VII. Residues of PCB*s in Human Tissue and Milk Yobs (1972) ' is reported that 31.1 percent of 637 samples of human adipose tissue collected from the general population as part of the Human Monitoring Survey during 1971 were positive for PCBs in measureable amounts. These samples were collected In 18 states HONS 056405 and the Dlatrlct of Columbia and poaitive sample* vara obtained froa aach of the sampling states and Dlatrlct. Tha percentage of PCB levels ranged from 34.2 percent non-detected, 33.3 percent lea* than 1 ppm, 27.3 percent 1-2 ppm and 5.2 percent greater than 5.2 ppm. Kutz and Straseman (1975) have deecrlbed tha reaulte of PCB monitoring during flacal year* 1973 and 1974 In which 35.1 percent and 40.3 percent, respectively, of the tleeuea collected contained levels of 1 ppm or more of PCBs on a net-veight basis. Analysis of the tlseue revealed that the compounds found In adipose tissue vers meet comparable to those prevalent In Arodor 1254 and Aroclor 1260. Additional analysis Indicated that tha most frequently encountered PCB residues vere petan-, here-, and heptachloroblphenyl compounds. Residues of PCBs have also been detected In a study of human milk collected In Colorado where 8 or 40 samples contained realduee ranging from 40 to 100 ppb (Savage etal., 1973). A study of adipose tissue samples collected at autopsy from Canadians (Prant at al., 1975) Indicates that tha majority of Canadians have adipose tissue residues of 1 to 2 mg/kg of PCB. All adipose tissues had datectabla levels of PCBs and 30 parcent of tha sasiplss had PCB realduaa greater than 1 mg/kg with a range 135 WOWS 056406 of 0.11 to 6.60 mg/kg. PCB residues in human milk from Ontario residents were found to be approximately 1 mg/kg on a fat basis. 186 MCN$ 05640? HUMAN EXPOSURE TO THE POLYBROMINATED BIPHENYLS I. Introduction and Background The polybromlnated biphenyls (PBBs) In this report refers to either Flremsster BP-6 or hexabromoblphenyl manufactured by Michigan Chemical Corporation for usa aa a flame retardant for thermoplastics. This product la a mixture of bromlnated biphenyls with an average bromine content equivalent to about six bromine atoms per biphenyl molecule. Flremastsr BP-6 la a mixture of the following bromlnated biphenyls W): Tetrabromobiphenyl 2.OX Pentabrouoblphsnyl 10.6X Hexabromobiphenyl 62.81 Heptabroaobiphenyl 13.8X Other broooblphenyls 11.6Z The Michigan Chemical Corporation has stated that to their knowledge Flremsster BP-6 is the only polybromlnated biphenyl produced in consasrclal quantity in the U.S. Their production estimates are: Year 1970 1971 1972 1973 1974 (Projected) Pounds 20,000 200,000 2,300,000 3,900,000 4,800,000 187 HONS 056408 Firemaster BP-6 has been used as a flame retardant in the manufacture of typewriter, calculator and microfilm reader hous ings, radio and TV parts, miscellaneous small automotive parts and small parts for electrical applications. The use of Flremaater BP-6 has been restricted to those applications where the end-use product Is not exposed to either animal feed or food and there la no known use in flame retarding fabrics where human exposure would occur (1). The ultimate disposition of Firerooster BP-6 upon burial is uncertain. The Michigan Chemical Corporation has stated that in their opinion this material will eventually undergo oxidative/ biological degradation forming carbon dioxide, water and bromide ion (1). II. Human Exposure In October of 1973, Adverse health effects were observed in cattle in several dairy herds In the State of Michigan. At that time, the cattle refused to eat manufactured feed; milk production decreased; there was a loss in body weight and the cattle developed abnormal hoof growth with lameness; cattle and swine aborted; and farmers reported the inability to breed heifers after they consumed feed manufactured by Farm Bureau Services. A herd of some 100 head of cattle sent to : laughter during this time period, exhibited enlarged 1 Ivors. 168 HONS 056409 v- Analysis of samples of the suspected feed by laboratories of the U.S. Department of Agriculture at Beltsvllle, Maryland, revealed that the feed was contaminated with a flame retardant chemical* hexabromlnated biphenyl. Subsequent investigation revealed that the Michigan Chemical Corporation manufactured magnesium oxide* a dairy feed supplement sold under the tradename Nutrimaster* and a flame retardant, hexabromlnated biphenyl, sold under the tradename Flremeeter BP-6. Both of these products were distributed In brown paper bags with either the name Nutrimaeter or Flremaster stenciled across the top of the bag. Whan the top of the bag vaa torn off and discarded, Identification was essen tially lost. Aa the reault of a mix-up In bags, Flramastsr BP-6 was mixed with animal feed in place of the Nutrimaster* apparently in the same proportion of use for the Nutrimaster. It appears that thraa kinds of faad were initially involved in this episode with PBB levels as follows: Feed Mo. PBB i 405 2.4 410 1,790 407 4,300 Samples of milk collected from Individual farms soon after the PBB waa Identified es the contaminant ranged from 2.8 ppm on 189 MONS 056410 a fat basis to 270.5 ppm on a fat basis. Other products seized and destroyed included: Rutter Cheese Canned milk Range 1-2 ppm 1.4 - 15.0 ppm 1.15 - 1.62 ppm It has been estimated that between the onset of contamination in the fall of 1973 and the establishment of the quarantine of affected herds and flocks in the spring of 1974 over 10,000 Michigan residents have been exposed to PBB through the consumption of contaminated milk, meat and other dairy products. A considerable amount of variation In exposure has proabably occurred in both length of exposure and levels of exposure. As a group, the farm family members have been at greatest risk followed by those indi viduals who purchased dairy products from contaminated farms on a regular basis. In order to determine whether or not persons exposed to PBBcontaminated products had suffered any acute adverse health effects, the Michigan Department of Public Health undertook a series of studies in the summer and fall of 1974. Study participants for the exposed group were dairy farm residents from farms which had been quarantined by the Michigan Department of Agriculture. The exposed subjects were limited to those who had lived or worked on the quarantined dairy farms for more chan six months 9ince May of 1973. Non-expoeed subject were randomly selected from a Uet of dairy producers In the same geographical area where farms had not been quarantined. A total of 298 persons were interviewed In the study and physical examinations and/or blood samples were obtained for 110 persons in the exposed group and 104 persona from the control group. The Michigan Department of Public Health has reported (1975) that responaee to a set of 24 specific medical conditions revealed chat none of the health complaints occurred consistently In either of the study groups. Statistical analysis showed that none of the listed complaints was significantly more frequent In those persons with the highest PBB levels when compered to other study subjects. Physical examinations of adults and children shoved no unusual abnormalities of the heart, liver, spleen or nervous system. Urinalyses and complete blood counts did not reveal a significant excess of unusual abnormalities related to exposure or PBB levels. These studies shoved that blood levela of PBB were significantly hlghar In tha study subjects from quarantined farms at compared to thoee from tha non-quarantined farms; although soma suhjacts from the farms shoved low PBB blood levels (Table 18). Several exposed females delivered normal babies without complication. Teste shoved concentrations of PBB In breast milk to be considerably hlghar than that found in paired blood plasms (Table 19). 191 MONS 056412 Table IK Distribution of PBB Blood Levels, Michigan, 1974 PBB Blood Levels (ppm) 0 0.002 - 0.019 0.020 - 0.090 0.100 - 0.490 0.500 - 2.260 Quarantined Farms Adults No. % Children No. 7. -3 3.7 43 52. U e 19 23.2 10 U 13.4 3 6 7.3 7 _ 28.6 35.7 10.7 25.0 Nonquarantined Farms Adults Children No. % No. y. 21 28.4 _ 52 70.3 29 1 1.4 1 00 0 00 0 _ 96.7 3.3 0 0 TOTAL 82 100.0 28 100.0 74 100.1 30 100.0 Table 19 Comparison of PBB Concentrations In Human Breast Milk and Blood Plasma Dace PBB Levels (ppm) Breast Milk Blood Plasma 12/74 10.800 .082 6/74 22.700 .252 10/74 1.800 .014 3/75 .210 .003 3/75 92.660 1.06B Paired samples of adipose tissue and blood were collected In a group of 13 individuals entering the hospital for surgical procedures. The concentration of PBB in adipose tissue ranged from 61 to 370 times the PBB value found in blood plasma with nn average ratio of 175 to 1 (Table 20). 197 HONS 056413 TABLE 20 Comparison of FBB Concentration* In Human Adlpoie Tlssua and Blood Plasma Data 11/4 PBB Levels (PP) Adipose Tissue Blood Plasms .410 .002 6/74 - 1/75 1.400 .005 3/75 3.000 .012 3/75 174.000 1.068 3/75 .248 .002 3/75 .274 .003 4/75 .177 .003 4/75 .152 .003 4/75 1.140 .004 4/75 4/75 .808 .530 .004 .007 4/75 .210 .003 5/75 1.110 .003 Report* of health complaint* such as numbness, stomach pain, headache, fatigue and anxiety continue to be reported In the various near*paper* In Xichlgan. Reports have also appssrsd In ths prass that several physicians In Michigan hava raportad abnormal llvar function testa la patient* exposed to PBB. Attempts to verify these reports with physician* have baan unsuccessful. 191 HONS 056*14 Btche, C. A* and liak, D. J. (1973), A Qualitative Hithod for Detecting Hydroxylated Metabolite# of Polychlorinated Biphenyls Bull Environ Contaa Toxicol 9:315-317. Bailey, S., and Bunyan, P. J. (1972), Interpretation of Persistence and Effects of Polychlorinated Biphenyls In Birds, Nature 236:34-36. Bareotti, D. A., and Allen, J. R. (1975), Effects of Polychlorinated Biphenyls on Reproduction In the Primate, Fed Proc 34:338. Bastomsky, C. H. (1974), Effects of a Polychlorinated Biphenyl Mixture (Aroclor 1254) end DDT on Biliary Thyroxin Excretion In Rata, Endocrinol 95:1150-1155. Bastomsky, C. H., Solymoss, B., Zsigmond, G., and tfyse, J. M. (.1975), Mechanism of Polychlorinated Biphenyl-Induced Hypobiilrublnaemie, Clin Chin Acta 61:171-174. Bastomsky, C. H., and Wyse, J. M. (1975), Enhanced Thyroxine Metabolism Following Cutaneous Application of Microscope Ioneraion Oil, Rea Conaun Chen Pathol Pharmacol 10:725-733. Bauer, H. Schulz, K. H.f and Splegelberg, U. 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(1974a), The Findings of Chlorinated Dibenzofurans in a Japanese Polychlorinated Biphenyls Sample, Bull Envir Contain and Tox 12:338-342. Roach, J. A. C. and Pomerantz, I. H. (1974b), The Finding of Chlorinated Dibenzofurans in Aroclor PCBs of Recent Manufacture, Paper No. 53 presented at 88th Annual Meeting of Assn, of Official Anal. Chemists, Washington, DC, Oct 14-17. Ruzo, L., Zakik, M., and Schuetz, R. (1974), Photochemistry of Bioactive Compounds: Photoproducts and Kinetics of Poly chlorinated Biphenyls, J Agric Food Chem 22:199-202. Ruzo, L. and Zabik, M. (1975), Polyhalogenated Biphenyls: Photolysis of Hexabrotno and Hexachlorobiphenyls in Methanol Solution, Bull Environ Contact Toxicol 13:1B1-162. Safe, S., Hutzinger, 0., and Jones, D. (1975), The Mechanism of Chlorobiphenyl Metabolism, J Agric Food Chem 23(5):851-853. Safe, S., Ruzo, L. 0., Jones, D., Platonow, N. 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(1973), Collaborative Study of the Recovery and Cas Chromatographic Quantitation of Polychlorinated Biphenyls in Chicken Fat and Polychlorinated Biphenyl - DDT Combinations in Fish, JAOAC 56:1015-1023. Schmoldt, A., Benthe, H. F., and Fruehling, R. (1974), Induction of Rat liver Enzymes by Polychlorinated Biphenyls (PCBs) in Dependence on the Dose and Chlorine Content, Arch Toxicol 32:69-81. Schulte, E. and Acker, L. (1974), Identifizierung und Metabolisicr* barkeit von Polychlorierten Biphenylen, Natuwissenschaften 6_1: 79-50. Schwartz, L. (1943), An Outbreak of Halowax Acne ("Cable Rash") Among Electricians, JAMA 122(3) i 158-161. Schwartz, L., and Barlow, F. A. (1942), Chloracne from Cutting Oils, Pub Health Rpts 37(47):1747-1750. Schwartz, L. (1936), Dermatitis from*$ynthetic Resins and Waxes, Am J Pub Health 26:586-592. Schwet2, B. A., Norris, J, M., Sparschu, G. L. , Rowe, V. K., and Cehring, P. J. (1973), Toxicology of Chlorinated Dibenzo-pdioxins, Adv Chem Series 120:55-69. Schwetz, B. a., Keller, P. A., and Cehring, P. J. (1974), The Effect of Purified and Commercial Grade Pentachlorophenol on Rat Embryonal anci Fetal Development, Tox and AppI Pharm 28:151-161. Sharp, C. W., Hunt, D. G., Clements, S. T,, and Wilson, W. E. (1974), Influence of Dichlorodiphenyltrichloroethane, Poly chlorinated Biphrnyls and Anionic Amphiphilic Compounds on Stabiliziation of Sodium and Potassium Actlvitated Adenosine Triphosphatases by Acidic Phospholipids, Mol Pharmacol 10:119-129. Simon, N. and Sikloni, C. (1974), Influence of Environmental Factors on Porphyrin Metabolism, Berufs-Dermatosen 22:237-259. Sinclair, P. R., Cr.inick, S. (1974), Uroporphyrin Formation Induced by Chlorinated Hydrocarbons (Lindane, Polychlorinated Biphenyls, Tctrachlorodiben20-p-dioxin). Requirements for Endogenous Iron, Proten Synthesis, and Drug Metabolizing Activity, lllochcn Biophys Res Commun 61:124-133. HONS 056438 Sissons, D. and Welti, D. (1971), Structural Identification of Polychlorinated Biphenyls In Commercial Mixtures by Gas-Llquld Chromatography, Nuclear Magnetic Resonance and Mass Spectrometry, J. Chromatog. 60,:15-32. Sivalingan, P. M., Yoshida, T., and Inada, Y. (1973), Modes of Inhibitory Effects of Polychlorinated Biphenyls on Oxidative Phosphorylation of Mitochondria, Bull Environ Contain Toxicol 10:242-247. Smith, F. (1948), U. S. Patent 2,449,088; (1949), Chem Abstr 43: 813. Smith, L. W., Fries, G. F., and Welnland, B. T. (1973), Utilisation of Contaminated Dehydrated Poultry Excreta as Feed by Cove and Polychlorinated Biphenyl Residues in Milk, J Dairy Scl, in press. Smrek, A., Adams, S., Middle, J., and Kimbrough, R.: Pharmacody namics of Hirex in Goats: Effect of Reproduction and Lactation. Submitted to Arch Environ Health. Societe d'Dleccrorhimie, d'Electrometallurgie E+ dee Aclerlee Electriques d'Uglne (1962) Relglan Patent 613,066; (1962) Chem Abstr 57:16492. Squire, R, A. and Levitt, H. (1975), Report of Workshop on Classification of Specific Heaptocellular Lesions In Rats, Cancer Research 35:3214-3223. Stalling, 0. L. and Huckens, J. N. (1971), Gas-Liquid Chromatography - Mass Spectrometry Characterization of Poly chlorinated Biphenyl (Aroclora) and ^Cl-Labllng of Aroclors 1248 and 1254, JAOAC 54:801-807. Stalling, D. L. and Mayer, F. L. Jt. (1972), Toxicitiea of PCBs to Fish and Environmental Residues, Environmental Health Perspectives 1:159-164. Stalling, D. L. Ttndle, R. C., Johnson, J. L. (1972), Cleanup of Pesticide and Polychlorinated Biphenyl Residues In Fish Extracts by Gel Permeation Chromatography, JAOAC 55:32-38. Street, J. C. and Shams, K. P. (1975), Alteration of Induced Cellular and Humoral Immune Responds by Pesticides and Chemicals of F.nvlromncntal Concern: Quantitative Studies of Immunosuppression by ))DT Aroclor 1254, Carbary!, Cnrbofuran, and Methylparathlon, Tox Appl Phnrm 32:587-602 X0NS 056439 Street, J. C., Urry, F. M., Wagstaff, D. J., and Blau, A. D. (1969), Comparative Effects of Polychlorinated Biphenyls and Organochlorine Pesticides in Induction of Hepatic Microsomal Enzymes, 150th National Meeting, American Chemical Society, New York, NY. Sugiura, K., Haltorl, M., Baba, M., and Goto, M. (1975), Accumu lation and Excretion of PCBs in the Mouse, Chemosphere 4(3):181-168. Sundstrom, G., Hutzlnger, 0., and Safe, S. (1976), Identification of 2,2*,4,4',55'-Hexabroobiphenyl as the Major Component of Flame Retardant FireMaster^ BP-6, Chemosphere 5:11-14. Tas, A. C. and Kleipool, R. J. C. (1972), Characterization of the Components of Technically Polychlorinated Biphenyl Mixtures - II, Bull Envir Contaa and Tox 8,: 32-37. Trotter, W. (1975), Removing the Interference of DDT and Its Analogs In the Analysis for Residues of Polychlorlnatsd Biphenyls, JAOAC 58:461-465. Trotter, W. J. and Young, S. J. V. (1975), Limitation on the Use of Antimony Pentachloride for Perchlorlnetlon of Polychlorinated Biphenyls, JAOAC 58:466-468. Trotter, W. J. (1976), PCB Analysis of Rice Oil Associated with Yusho, Memo to Frank Cordle, FDA, March 11. Tsukamoto, H, et el. (1969), The Chemical Studies on Detection of Toxic Compounds In tha Rice Bran Oils Used by the Patients of Yusho, Fukuoka^ Acta Medics 60:496-512. Tucker, R. K., and Crabtree, D. G. (1970), Handbook of Toxicity of Pesticide* to Wildlife, Bureau to Sport, Fisheries, and Wildlife, Resources Publ. 84, U.S. Dept* of Interior, Washington D. C. Turner, J. C., and Green, R. S. (1974), Effect of a Polychlorinated Biphenyl (Aroclor 1254) on Liver Microsomal Enzymes In the Male Rat, Bull Environ Contaa Toxicol 12:669-671. Valnlo, H. (19/4), Enhancement of Microsomal Drug Oxidation and Glucuronldatlon in Rat Llvsr by an Environmental Chemical, Polychlorinated Biphenyl, (Them - Biol Interact 9:379-387. Van Miller, J. P., Hsu, I. C., and Allen, J. R. (1975), Distri bution and Metabolism of ^H-2,5,2',5-tatra-chlorobiphenyl in Rats, Froc Soc Exp Biol Med 148:682-687. HONS 056440 Veith, C. D, (1975), Baseline Concentrations of Polychlorinated Biphenyls and DDT in Lake Michigan Fish, 1971, Pest Monitoring _ Journal 921-29. Villeneuve, D. C. Grant, D. L., Phillips, W. E. J., Clark, M. L., and Clegg, D. J. (1971a), Effects of PCS (Polychlorinated Biphenyl: Aroclors 1221 and 1254), Administration on Microsomal Enzyme Activity In Pregnant Rabbits, Bull Environ Contain Toxicol 6:120-126. Villeneuve, D. C., Grant, D. L., and Phillips, W, E. J. (1972), Modification of Pentobarbital Sleeping Times In Rats Following Chronic PCB ingestion, Bull Environ Contam Toxicol 7:264-269. Villanueva, E. C., Jennings, R. W., Burse, V. W., and Kimbrough, R. D. (1974), Evidence of Chlorodlbenzo-p-dioxln and Chlorodibenzofuran in Hexachlorobenzene, J Agr Food Chem 22:916-917, Villeneuve, D, C., Grant, D. L., Khera, K., Clegg, D. J., Baer, H., and Phillips, W. E. J. (1971b), The Fetotoxicity of a Polychlorinated Biphenyl Mixture (Arocior 1254) in the Rabbit and in tl>e Rat, Env Physiol 1^:67-71. Vos, J. G. and Koeman, j. H. (1970), Comparative Toxicologic Study with Polychlorinated Blphenyla in Chicken with Special Reference to Porphyria, Edema Formation, Liver Necrosis, and Tissue Residues, Tox Appl Pharm 17,:656-668. Vos, J. C., Koeman, J. J., Van der Maas, H. L.f Ten Noever de Brauw, M. C., and De Vox, R. H. (1970), Identification and Toxicological Evaluation of Chlorinated Dibenzofuran and Chlorinated Naphthalene in Two Commercial Polychlorinated Biphenyls, Fd Cosmet Toxicol 8:625-633. Vos, J. G. and Beems, R. B. (1971), Dermal Toxicity Studies of Technical Polychlorinated Biphenyls and Fractions Thereof in Rabbits, Tox Appl Pharm 19:617-633. Vos, J. C. (1971), Polychlorinated Biphenyls as Inducers of Hepatic Porphyria in Japanese Ounil with Special Reference to -Amlnoleyullnic Acid Synthesase Activity, Fluorescence, Acid Residues in Liver, Toxicol Appl Pharmacol 20:232-240. Voa, J. C, and Notenboom-Ram, E. (1972), Comparative Toxicity Study of 2,4,5,2',4*,5'-Hexachlorobiphenyl and a Polychlorinated Biphenyl Mixture in Rabbits, Tqxt and Appl Pharm 23:563-578. HONS 056441 Vo, J. G. (1972), Toxicology of PCB's for Mammals and for Birds, Env Hlth Persp 1:105-117. Vo, J, C. and VanGenderen, H. (1973), Toxicological Aspects of Itnoumo-euppression. Aspects of Immunosuppression Published In Pesticides in the Environment, a Continuing Controversy, Ed. by W, B. Deichman, (8th International Conference on Toxicology and Occupational Medicine, Miami), Published by Intercontinental Med. Book. Co., New York, NY. Voa, J. G. and DeRoij, T. 1. (1972), Immunosuppressive Activity of a Polychlorinated Biphenyl Preparation on the Humoral Immune Response in Guinea Pigs, Tox Appl Pharm 21:549-555. Walker, C. R. (1975), Pre-1972 Knowledge of Non Human Effects of Polychlorinated Biphenyls, Nat Conf on Polychlorinated Biphenyls, Chicago, Illinois. Washerman, D., Wasserman, M. (1972), Ultrastructure of Adrenal Zona Fasciculata In Polychlorinated Biphenyls Receiving Rats, 17th Intern Congress Occ Health, Buenos Aires. Wasserman, D., Wasserman, M., Cucos, S., and Djavaherlan, M. (1973), Function of Adrenal Glad-sone Fasciculata in Rats Receiving Polychlorinated Biphenyls, Environ Res 6:334-338. Webb, R. G. and McCall A. C, (1972), Identities of Polychlorinated Biphenyl Isomers in Aroclors, JAQAC 55:746-752. Webb, R. G. and McCall, A. C. (1973), Quantitative PCB Standards for Electron Capture Gaa Chromatography, J Chromatogr Sci 11:366-373. Welngarten, H. (1961), Steric Effects in the Gooberg Reaction, J Org Chem 26730-733. Welborn, J, A., Senator and Chairman, A Report from the State Senate Special Investigating Committee, The Contamination Crisis in Michigan, July, 1975. WHO Task Croup (1975), Environmental Health Criter for Poly chlorinated Biphenyls and Terphenyls. - Wit, J. G. (1972), Enzyme Induction with Specific Reference to Chemical Porphyria, Third Technical Meeting on Occurrence and Significance of Chemicals in the Environment, Berlin, Germany. HONS 056442 Wong, R. G., Novicki, H. C., and Norman, A. W. (1974), Effect* of Polychlorinated Biphenyls on Calciferol (Vitamin D) Mediated Calcium Metabolism, Peadc Biochem Phvalol 4*170-177. Yamamoto, H., and Yoshlaura, H. (1973), Metabolic Studies on PCBs. III. Complete Structure and Acute Toxicity of the Metabolites of 2,4,3,4*-tetrachlorobiphenyl, Chem Pham Bull 2112237-2242. ----------------------------- Yamamoto, H. (1974), Statistics of Occupational Disease (Result of Speclol Medical Examination), New Occupational Health Handbook 1010-1013, The Institute for Science of Labour, In Japanese. Yap, K. H., Deuaiah, D., Cutkomp, L. K., and Koch, R. B. (1971), Sensitivity of Fish atpase to Polychlorinated Biphenyls, Nature (London) 233:61-62. Yobs, A. R. (1972), Levels of Polychlorinated Biphenyls in Adipose Tissue of tha General Population of the Nation, Environ Health Perspectives ,1:79-81. Yushlnura, T. (1971), Epidemiological Analysis of "Yusho" Patients with Special Reference to Sex, Age, Clinical Grades and Oil Consumption, Fukuoka, Acta Modica 62:104-108, Young, S. and Burka, J. (1972), Micro Scale Alkali Treatment for Use in Pesticide Residue Confirmation end Sample Cleanup, Bull Environ Contaa Toxicol 7J160-167. Zepp, R. L. Jr., Senders, O. T., and Kirkpatrick, R. L. (1974), Reduction of Pentobarbital-Induced Sleeping Tinea In PCB (Poly chlorinated BiphenyD-Treeted Cottontail Rabbits, Bull Environ Contam Toxicol 12:518-521. Zitko, V,, Hutzlnger, O., and Choi, P. M. K. (1972), Contamination of the Bay of Fundy - Gulf of Maine Area with PCBs, PCTs, Chlorinated DBF and DBD, Environ Health Parspect 1^47-50. Zitko, V., Hutzlnger, O., and Safe, S. (1971), Retention Times and Electron Capture Detector Responses of Some Individual Cbloroblphenyls, Bull Environ Contam and Toxicol 6:160-163. Zitko, V., Hutzlnger, O., and Safe, S. (1971), Retention Times and Electron-capture Detector Responses of Some Individual ChloroblphenyIs. Bull Environ Contam ToxlcoJ 6:160-163. Zitko, V., Hutzlnger, 0., and Choi, 1*. M. K. (1974), Determination of Pentachlorophenol and Chloroblphenylola In Biological Samples, Bull Environ Contam Toxicol 12:649-653. M0NS 056<i<i3 NAME Chlorlnetod biphenyls Appendix A CHLORINATED AROMATIC COMPOUNDS REFERRED TO IN CHEMISTPT REPORT STRUCTURE EXTENT Or CNLORIHATION NU?fBER OF x- 1-10 209 - Chlorinated naphthalenes \-l-8 75 Chlorinated dibeniofurans n*l$ 135 Chlorinated dibo n ze-p-'dloxi ns pantuchlorophenol hcxachlorobemen* Cl Cl Cl ci Cl 75 MOMS 058444 Attachment 2-1 Calculation of dosages received by monkeys in reference 16 The amount of food eaten in relation to body weight varies between different animal species, including man. To enable comparisons, food or food component intake frequently is standardized by expressing the amount eaten as a function of body weight. The latter is known as the dosage, while the amount eaten would be the dose. Table 1 lists mean body weights for all experimental animals. These range from 5.57 kg pretest to 4.84 kg at the end of 5 months on test. An overall mean body weight of 5.32 kg. was estimated by averaging the six figures listed. Table 3 shows total 6-month polychlorinated biphenyl intake as 85,29 mg for 2.5 ppm group and 175.36 mg for the 5.0 ppm group. Average daily intake was calculated by use of the following formula: pg/kg/day total mg intake x 1_____ x 1000 pq r body wt. (kg) 18C days mg At 2.5 ppm, the dosage was approximately 89 pg/kg/day At 5.0 ppm, the dosage was approximately 183 pg/kg/day For comparison, FDA estimated safe intake as 1 pg/kg/day (14) Thus, even though the monkey dietary levels of 5.0 ppm and 2.5 ppm were described as levels "equal to, and 50% of, the concentrations allowed in certain foods destined for human consumption" (16), it appears that the monkeys actually received about one hundred and two hundred time! the dosages which FDA estimated as safe from the human experience at yusho. Attachment 2-1 MONS 056445 EVALUATION OF CHANGES OF THE LEVEL OF POLYCHLORINATED BIPHENYLS (PCB) IN HUMAN TISSUE Final Report on FDA Contract 223-73-2209 Michigan Department of Public Health Lansing, Michigan Harold E. B. Humphrey, Ph.D. Project Director /'j/7 -m~30$7 - r. Attachment 2-7 1 HONS 05 b't'tb EVALUATION OF CHANGES OF THE LEVEL OF POLYCHLORINATED DIPHENYL IN HUMAN TISSUE SUMMARY OF riNDINGS A two year study was made of persons who regularly consumed Lake Michigan sport fish and randomly selected persons who did not consume such f.ish. A total of 161 adults from the shoreline communities of Traverse City, Manistee, Ludington, and South Haven participated. 105 of these participants consumed more fish than recommended by the Michigan Department of Public Health (no more than one meal per week or 24 pounds per year) and constituted the PCB "exposed" group. The 37 participants eating six pounds or less per year, 19 eating an intermediate amount, and 19 fish eaters from Algonac where sport caught fish are low in PCB were used for comparison groups. A short medical history, a dietary record, and blood specimens were obtained for all participants. The following conclusions were reached from these data: 1. PCB blood levels ranged from 0.007 ppm in persons who ate no fish to a maximum of 0.366 ppm fcr a person eating Lake Michigan fish. 2. There was a direct relationship between the size and quantity of sport species of fish eaten and the PCB levels found in human blood. 3. In 1973, thase who annually consumed 2>i or more pounds of sport fish from Lake Michigan had a mean blood PCB value of 0.073 ppm; persons annually eating H pounds or less had a mean level of 0.020 ppm; persons who ate no fish averaged 0.017 ppm; and persons eating 24 or more pounds annually from Lake St. Clair had an average blood level of 0.023 ppr.. The PCB blcod levels observed for the Lake Michigan study groups were similar in 1979. HONS 056447 4. The level of PCB found in the blood of participants did not change significantly from year to year, nor did it diminish significantly when fish consumption was eliminated for up to nine months. 5. The PCB levels observed in cooked fish were lower than expected from reported levels in raw fish. 6. Eighty-two percent of those who exceeded the state's - recommended maximum intake (no more than one meal per week or 24 pounds per year) of Lake Michigan fish received an estimated annual dose of PCB which was greater than that cited by the Food and Drug Administration as the maximum annual dose of PCB which should be received over a protracted period of time. This estimated intake ranged from 0.49 pg PCB/Kg body weight/day to 3.94 pg PCB/Kg body weight/day and averaged 1.7 pg/Kg body weight/day. The Food and Drug Administration recommends that PCB intake not exceed 1 pg/Kg body weight/day for long term exposure. 7. As a group, the exposed participants had no health problems or medical conditions that could be correlated with PCB blood levels, exposure to Lake Michigan fish, or known symptoms of PCB poisoning. CONCLUSION: Consumption of Lake Michigan fish contributed substantially to the PCB levels in the blood of humans. The dose received is insufficient to produce a detectable effect on human health at present. It has not been determined whether long term exposure to PCB contaminated fish will result in a continuing accumulation of PCB's or eventually produce identifiable health effects in humans. The data gathered to date appear to justify continued surveillance of this situation in Michigan. The data also justify a continued recosmendation that intake of Lake Michigan salmon and lake trout be linilcd to less than one meal per week or 24 pounds per year. MOMS 056446 TABLE OF CONTENTS INTRODUCTION .................................................................................................. 1 PCB PROBLEM IN MICHIGAN ....................................................... . 3 THE MICHIGAN PCB STUDY.............................................................................. 6 rield Survey Design .................................................................... 8 Study Findings .......................................................... 14 Analytical Methodology ........................................................... 47 Chosen Analytical Procedures ............................................ SI Special Tests ................................................................................... 54 APPENDIX A..........................................................................................................60 APPENDIX B..........................................................................................................66 APPENDIX C.......................................................................................................... 68 APPENDIX D . . . ...................................................................... 75 APPENDIX E...................................................................................................... 7 9 BIBLIOGRAPHY .................................................................................................. 03 1 MONS 056449 LIST or TABLES Humber 1 2 3 4 5 B 7 8 9 10 11 12 Title Page PCB Contamination in Whole Fish............................................. 5 1974 Catch and Effort Report for Lake Michigan . . 7 FCB Study, Participation by City...........................................12 Correlation of Fish Exposure with FCB Values in Blood Over a Two Year Period, Traverse City Group..........................................................................................................18 Correlation of Fish .Exposure with PCB Values in Blood Over a Two Year Period, Ludington Group..........................................................................................................20 Correlation of Fish Exposure with PCB Values in Bicod Over a Two Year Period, Manistee Group..........................................................................................................21 Correlation of Fish Exposure with PCB Values in Blood, SouthHaven and AlgonacGroups ...................... 22 Year to Year Comparison of Mean PCB Values by City......................................................................... 27 Evaluation of Mean PCB Values, Control Group ... 28 Correlation of PCB Levels in Humans and Length of Abstinence From FishConsumption......................................30 PCB Levels in Cooked Fish Consumed by Humans ... 33 Cooked Fish Survey, Mean PCB Concentrations by Conmunity........................................................................................... 34 ii HONS 056450 LIST OF TABLES (continued) Number 13 1M 15 16 17 18 19 20 Title Page Estimated TCB Ingestion rrom Fish, Annual Dose and Mean Blood Values for Persons Consuming 2M or More Pounds per Year........................ 36 Calculated PCB Ingestion and Dose From Consumption of Great LakesFish .......................................... M2 Correlation of Blood PCB Levels with Number of Years of Fish Consumption.................................................... 43 Exposed and Control Participant Responses to Health Conditions ......................................................................... MS Mean and Standard Deviations of PCB Values and Sample Sizes for Each Condition by Reporting Status ..................................................................................................... M6 Best Specimen Comparison...........................................................59 Blood Samples Tested by the DegradationDerivatization Technique ........................................................... 67 Participant Physical Data.................................. 69 ii i HONS 056451 LIST OF FIGURES Number 1 2 3 4 5 6 7 8 9 10 Title Page Location of PCB Exposed 6 Control Participants . . 9 Frequency Distribution of Reported Great Lakes Fish Consumption, Exposed Croup, Traverse City, Manistee Ludington........................ ...........................................16 Correlation of PCB Levels in Human Blood with Fish Consumption................................................................................. 24 Scatter Plot of Pounds of Fisli Consumed v s the Natural Log Of PCB bevels in Human Blood........................25 Correlation of PCB Levels in Human Blood with PCB Ingestion From Fish, Traverse City Group ... 40 Correlation of PCB Levels in Human Blood with PCB Ingestion From Fish, Manistee, Ludington, and South HavenGroups .................................................................. 41 Analytical Precision, Plot of PCB Values for Replicate Tests ........................................................................... 56 Chromatograms for, (A) 1.5 ng Aroclor 1254 and (B) 1.34 mg Cooking Oil....................................... . . . . 57 Chromatogram for Aroclor 1260 (0.6 ng).............................58 Change in Blood PCB Levels Over Time for Persons Exposed toLake Michigan Fish............................ 77 iv HONS 056452 LIST or FIGURES (continued) Number 11 12 13 Title Page Change in Blood PCB Levels Over Time for Persons Exposed to Lake Michigan Fish...........................78 Change in Blood PCB Levels Over Time for Persons Who Abstained From Lake Michigan Fish Consumption................................................................................. 81 Change in Blood PCB Levels Over Time for Persons Who Abstained From Lake Michigan Fish Consumption ..................................................... 82 MONS 056453 EVALUATION OF CHANGES OF THE LEVEL OF POLYCHLORINATED BIPHENYL IN HUMAN TISSUE INTRODUCTION Polychlorinated biphenyls (PCB) have been manufactured and used for over forty years in a wide variety of industrial applications which require a substance with chemical and thermal stability. Approximately 40% of these uses may have resulted in loss of PCB to the environment. PCB residues began to be identified in animal and environmental samples in the late 1960's and it was soon confirmed that this chemical was a persistent and a ubiquitous world wide environmental contaminant (1, 2). PCB's have been detected in manufactured products, air, water, snow and sediment samples as well as aquatic biota and terrestrial animals and birds. A consequence of this situation has been concern about the indirect contamination of some foods destined for human consumption and the resulting exposure of humans to PCB. Reports of the detection of low levels of PCB in tissues from the general population (3, 4) have confirmed that human exposure is a reality. The human health effects from PCB exposure, especially long term low level exposure, are not clearly understood. Information on PCB exposure for humans has been chiefly confined to acute effects from industrial exposure and an extensive food contamination episode in Japan where a disease called Yusho has been described (7). Although reports on the toxic symptoms observed with Yusho have generally been accepted as attributable to PCB poisoning, the suspected existence of other toxic contaminants in Japanese PCB preparation has raised questions on the cause of some of these observed effects. . MOMS 056454 ^Scientific information on the characteristic!) and toxicity of PCB's was reviewed, in 1972 (2 ), and currently by the Food and Drug Administration (FDA) with respect to the adequacy of the temporary tolerances for PCB (9). The body of data reported to date indicates that the toxicity of PCB varies with the extent of chlorination of the biphenyl molecule. These compounds are poorly metabolized and tend to accumulate and be retained in the lipid portion of animal tissues for prolonged periods (5, 10, 11). The accumulation and storage of a chemical would theoretically make it available, under appropriate conditions, for recirculation within the body allowing continuous or repeated exposure of target organs long after the original dose has ceased or when multiple low doses are received and accumulated to a critical level. The significance of the observed accumulation and storage of PCB in tissue is not fully known. However, recent reports of the continued presence of some clinical signs in Yusho patients years after the incident (5, 12) and the detection of significant levels of PCB in rat tissues ten months after exposure (11) tend to support the storage continued exposure contention. Moreover, recent studies reporting acute health effects including reproductive failure in primates fed low levels of PCB (13, 19), the induction of liver tumors in rats from PCB ingestion over prolonged periods (15), and the partial conversion of tetrachlorobiphenyl to a more acutely toxic monohydroxy-metabolite in rats (16) suggests that long term or low level exposure to PCB or its metabolic byproducts is not fully understood and may be of significance. The existing body of toxicity data for PCB has been sufficient for the FDA to regard these compounds as potentially serious health hazards in food. Temporary tolerances for I'CU's wore established by the FDA in 1973 (8). The determination of tolerance levels was predicated on the assumption that PCB exposure from food was sporadic and would diminish with time. Subsequent surveillance data has confirmed this to bo true 2 MONS 056A55 except for fresh water fish found in certain areas of the nation (9)., It has been concluded that frequent consumption of fish containing PCB in excess of 5 parts per million (ppm) poses a risk to public health. Data for the evaluation of this risk to humans has not been available. Concern about the risk and the need to obtain information on humans exposed to PCB's from consumption of contaminated fish was the basis for establishing the research contract reported "below. THE PCB PROBLEM IN MICHIGAN The Great Lakes surrounding Michigan represent one of the areas of the nation contaminated with PCB. The Michigan Water Resources Commission has reported that many surface water samples contained PCB at concentrations above the detection limit of 10 parts per trillion (ppt). Significant levels of PCB have been found in rivers and streams discharging into the Great Lakes. Effluents from wastewater treatment plants servicing industrialized communities have been found to be highly contaminated. Although efforts have been made to eliminate point source industrial losses of PCB, recent monitoring reveals that PCB contamination continues to be a problem. It has been suggested that highly contaminated bottom sediments in sewers and receiving streams may represent a reservoir for the continued release of PCB. It has been concluded that PCB continues to tie discharged into the Great Lakes from many small but diffuse sources including atmospheric fallout. PCB's were first detected in Great Lakes fish in 1969 warn substances interfering with the analysis of DDT in Coho salmon sampler, were determined to be PCB (3). Subsequent environmental monitoring has identified 3 HONS 056<|56 PCD contamination in fish from all of the Great Lakes. The problem is most severe in Lake Michigan and least severe in lakes Huron (except for Saginaw Bay) and Lake St. Clair. Concern for PCB contamination has focused on Lake Michigan because of the large investment in the rejuvenation of this lake for sport fishing through fish stocking programs. Statistical sampling surveys of contaminants in Great Lakes fish began in 1972. This program has shown that most species tested contained detectable levels of PCB's and that contamination levels were generally proportional to fish size (age) and highest in the predator species. Except for whitefish, the species of commercial or sport interest (trouts and salmons) from Lake Michigan were found to be most highly contaminated with PCB. Moderate and larger sized specimens have been found to "be well in excess of the PDA tolerance level of 5 ppm. A portion of the data obtained for fillets of lake trout from Lake Michigan during the 1979 Great Lakes Environmental Contaminants Survey is exemplified below: Lake Zone (Size) Pounds No. Fish Mean PCB (ppm) MM9 5-10 3 3.06 MMS 5-10 7 9.01 MM 6 5-10 . 20 9.08 MM7 5-10 11 11.86 MM8 5-10 6 11.93' The lake zones refer to the statistical sampling areas of Lake Michigan from which the specimens were taken and are identified in figure 1. The values shown indicate a trend Of increasing 'C.B levels as samples arc obtained further south in the luke along more heavily populated and industri alized areas of the State. M MONS 056457 Unfortunately PCB contamination of fish in the Great Lakes has not decreased to date. Table 1 shows data from a three-year study of two species of Lake Michigan fish by the U.S. Fish and Wildlife Service Laboratory. As shown, the PCB levels in lake trout appear to have risen and have not declined in Coho salmon over the study period. TABLE 1. PCB CONTAMINATION IN WHOLE FISH* Collection Length Sample Mean PCB Species E Location Year (inches) Size Value (ppm) Lake Trout near South Haven 1972 1973 . 1974 20 - 20 20 - 28 20 - 28 9 12.86 t 4.75 30 18.93 t 2.08 30 22.91 i 3.75 Coho Salmon near l.udington 1972 1973 1974 20 - 32 20 - 32 20 - 32 10 10.93 t 2.12 29 12.17 1 0.77 30 10.45 i 0.97 ftData from the Great Lakes Fishery Laboratory, U.S. Bureau of Sport Fisheries ard Wildlife. The continued discharge of PCB into the' water and trend analyses such as these indicate that PCB contamination of Great Lakes fish is a long term problem. Sport fishing represents an uncontrolled source of PCB contaminated fish for distribution and human consumptior . The State and federal inspection system controls exposure to PCB from commercial sources by seizure of contaminated fish before it reaches the market. Sport fishermen and those who consume sport caught Great Lakes trout and salmon represent the population most likely to be at risk to significant PCB exposure from fish. The Michigan Department MONS 056458 TABLE 2. 197M CATCH AND EFFORT REPORT FOR LAKE MICHIGAN Lake Trout SPECIES Rainbow - Coho Steel head Brown Trout Salmon Chinook Sa1mon total No. Caught/ Year Mean Weight/ F i sh Pounds of f1sh Caught/ Year . No. Pounds/ Fisherman/ Year 739,260 271,980 96.480 502.200 264,420 1,874,320 5-6 8-10 ' 5 6 15 - 4,065,930 2, <1*47.820 482,600 3,013,200 3,966,300 3,975,650 10.6 6.4 1.3 7.9 10.4 3 >.6 * Data from the Michigan Department of Natural Resources 197fi Annual Sport Fishing Survey. Information obtained from statistical quarterly census of the 381,600 licensed fishermen In the 18 Michigan counties bordering Lake Michigan. G HONS 056459 Natural Resources indicates that 381,000 licensed fishermen reside in the 18 counties bordering Lake Michigan. That department's 1978 sport fishing survey (Table 2) shows that this group catches about 18 million pounds of trout and salmon from Lake Michigan annually. As indicated, the average size of the fish being caught fall into the range which is significantly contaminated with PCB. Statistically, each of these fishermen obtains 36 pounds per year (3/8 pounds per week)of Lake Michigan fish for consumption. This is over three times the reported national per capita consumption of fish from commercial sources (11.8 pounds) but about half that reported for the Japanese (67.6 pounds) (Source: Production Yearbook, 1970, vol. 28, Pood and Agriculture Organization of the United Nations). Although the habits of individuals will vary it appears that as a group these sport fishermen consume more fish than the average and are successfully catching the PCB contaminated species from Lake Michigan. THE MICHIGAN PCB STUDY The !'ood and Drug Administration Contract 223-73-2209 with the Michigan Department of Public Health represents an attempt to evaluate human exposure to PCB from consumption of sport caught Lake Michigan fish. The study was principally designed to provide information on the levels and changes in the level of PCB's in humans. It was not intended to be a full toxicological evaluation of the human health effects of PCB exposure. The major objectives of the study were: 1, To conduct research on the analysis of PCB's in order to develop a sensitive laboratory method for detecting PCB concentrations in human blood. 2. To identify exposed and control population groups in selected geographical locations where PCB levels in fish are excessive. 7 HONS 056460 3. To select participants for the study and collect suitable blood specimens using accepted epidemiologic techniques. <i. To evaluate the concentration and variation of PCB's in representative groups of humans. 5. To estimate the fish consumption characteristics of the selected exposed group. 6. To determine the PCB levels in fish being consumed by humans. 7. To determine by laboratory analysis the concentration of PCB's in participants' blood specimens in order to evaluate A. The PCB ingestion frpm fish B. The PCB levels in the blood of humans who eat fish and who do not eat fish C. The variation of PCB levels in blood due to seasonal fish consumption habits D. The metabolic half-life of PCB's present in the blood of humans 8. To attempt to correlate levels of PCB in tissues with levels of PCB ingestion. IT CLP SURVEY DESIGN: The greatest opportunity for exposure to PCB's,as discussed above,was believed to lie in the consumption of certain sport caught species of Lake Michigan fish. Persons from four Michigan communities, South Haven, l.udington, Maniste .iikI Traverse City, all on the shoreline of Lake Michigan (sec figure 1) wore selected for the study. These communities are points of origin for Great Lakes sport fishing and span the principal areas of Lake Michigan where l'CB contaminated HONS 056461 9 MONS 056*62 fish have been identified. In addition, AJgonac, a shoreline community on Lake St. Clair where l'CB levels in fish are not excessive was included for a comparison of fishermen from a different geographical location. Exposed and control subjects for the study were personally selected by field workers following introductory publicity in the local news media of each community. Adults (age 18 and above) who caught and regularly ate sport fish from Lake Michigan were sought and encouraged to participate. Exposed participants, those consuming at least 24-26 pounds of Great Lakes fish per year (an annual average of at least one 8 oz. meal per week) were identified through a voluntary and referral system. Control participants, those consuming less than six pounds of Great Lakes fish per year (an annual average of zero to no more than one 8 oz. meal per month) were identified by contacting households randomly selected from the directory for each community. All potential participants, whether voluntary or randomly selected, were visited and personally interviewed in order to determine their eligibility for the study utilizing the above fish consumption criteria and a questionnaire developed for the study. Those judged eligible by the interviewer, as determined by the responses to Die fish consumption questions, were invited to participate in the study. Persons in agreement were then briefed on the purpose and extent of the s.tudy, were fully interviewed, given a bound log for recording their subsequent consumption of Great Lakes fish, and were asked to freeze portions of their fish meals. In each case the interviewer explained the importance and method for these procedures. Periodic contacts were made in order to maintain participant interest in the project ani to schedule blood sample collections. Specimen collections for laboratory analysis were scheduled at the local health department offices at tines convenient for the participants. Whole blood samples (20 cc) were collected by venipuncture by the 10 HONS 056i 63 Michigan Department of Public Health staff. During these appointments, questionnaire information was updated and an informed consent statement was signed by the participant. Exposed and control populations in South Haven, Ludington, Manistee and the Traverse City area were identified and selected for the study as described above. Blood specimens were collected from 1S2 persons during the Fall of 1973 at the end of the fishing season on the Great Lakes. This sampling provided a baseline for evaluation of PCB values after consuming sport fish through a whole fishing season. The classification and location of these participants are shown in Table 3. The representative number of randomly selected unexposed controls to be sought from each community was determined by consultation with statisticians. The group classified as "intermediate" represents persons who qualified as fish eaters when initially interviewed, but failed to actually consume a sufficient quantity of Groat Lakes fish during the 1973 season or the 1974-75 season to be classified as exposed fish eaters. Blood specimens were collected during the Spring of 1974 (April sample, Table 4) from seventeen of the Traverse City area fish eaters who had been sampled the previous Fall. These collections were made prior to the resumption of sport fishing for the 1974 season and represent an attempt to evaluate the seasonal variation in the level of PCB's in exposed persons. Consumption of frozen'or canned Great Lakes fish during the winter months represents a problem for the evaluation of seasonal variation. It was originally believed that consumption o.:' sport fish would cease shortly after the fresh supply was exhausted at the end of the fishing season. However, work with the study participants revealed that fresh caught sport fish are preserved, frozen or canned, to a much greater extent than expected. Thus, the cessation of fish consumption is not 11 MOMS 056464 TABLE 3 . PCB STUDY, PARTICIPATION BY CITY CITY Number Participants 1973 Number Participants 197^4-75 Total Number Par11c1 pants Over Life of Study Inter Inter 9 Indi Se K Med k Total Control mediate Exposed Total Control mediate Exposed viduals M F Age Traverse City 53 19 Manistee 21 7 ludlngton 28 10 South Haven 31 2 Algonac 1$ 0 5 29 60 13 10 37 74 32 42 49.! 2 12 21 5 3 13 25 13 12 45.! 8 10 20 7 5 8 33 15 18 41.( 4 25 31 21 10 42.! 5 14* 19 13 6 51. TOTALS 152 38 24 25 18 58 ,82 38 45.: 'Exposure hare constitutes consumption of fish from Lake St. Clair where PCB levels arc relatively low. 12 MONS 056465 abrupt but instead tapers off slowly as the preserved supply is exhausted or continues at a reduced level until the following fishing season. The 101 participants for the second <1974-75) phase of the study are listed in Table 3 by location. The July 1971 specimen collection (Tables 4, 5, G) provided a second baseline sample for the study. A portion of these participants are persons who were included in the Fall screening from the previous year (1973). This provided for a year to year comparison of PCB levels in certain individuals. The balance of the 58 fish eaters were recruited during the Spring of 1974. Emphasis was placed on recruiting persons with a history and expectation of catching and consuming an amount of fresh fish well- in excess of the required 24-26 pounds per year. This effort was in response to the realization that the quantity of fish consumption among Great Lakes fishermen is not consistent from year to year. Fish consumption appears to be dependent on the weather (number of trips onto the lake) and on success once at the fishing grounds. One of the problems encountered during this study was the inability of some exposed participants to catch and consume the anticipated quantity of fresh fish throughout the study. In some cases this necessitated a downward reclassificatioq of the participant. The 1974-75 phase of the PCF Project provided a second year comparison, a close monitoring of individual fish consumption over an entire fishirg season, and an evaluation of the change in level of FCB's in blood through a series of specimen collections after the voluntary termination o: fish consumption at the end of the season. Completion of the abstinence study was complicated by the staggered dates when individuals concluded one fishing season and began the next. In addition, the request for participants to cease eating sport fish represented a 13 MOMS 056466 break in their normal habit of consuming frozen fish through the winter. These problems resulted in the elimination of some participants from this portion of the study, because they failed to abstain from eating fish for a sufficient period of time. STUDY FINDINGS: The study was designed to include 140 to 160 participants. This goal was achieved as 182 persons participated during the 2 years of the study. Table 3 shows the classification of participants in each community for the 1973 and 1974-75 phases of the study. The size of the control group was based on the number needed to be statistically acceptable for data evaluation. The intermediate exposure classification was necessitated by the realization that a difference existed between estimated fish consumption based on recall responses to the questionnaire and actual consumption of fish during the study period. As explained previously some persons who were alleged to consume over 24-26 pounds of Lake Michigan fish annually failed to do so and were reclassified from the exposed to an intermediate (7 to 23 pounds per year) group. Many of the first year participants continued in the study to its conclusion. Those who elected to drop out and those who were reclassified as intermediate exposure after the first year were replaced by additional exposed participants for the second year of the study. Table 3 also shows that the distribution of males and females was nearly equal ard that the median participant age was about 4$ years. The age, sex, height, and weight data for each participant is provided in Appendix C. 14 MOMS 05646? The quantity of fish consumed by individuals in the exposed classification was as a group unimpressive but correlated with the calculated mean catch as reported in the sport fishing survey (Table 2). The reported popularity and catch success of sport fishing on hake Michigan and the deliberate effort to enlist reputed fishing enthusiasts in the study led to expectations that the exposed group would consume large quantities of fish. However, a plot of the distribution of mean annual fish consumption over the two fishing seasons of the study shows that the most frequently recorded quantity of fish actually consumed was in the 24-35 pounds per year range (Figure 2). The number of participants reporting a consumption of greater than 53 pounds was low. The highest reported consumption over the two year period was 180 pounds per year and the highest single season consumption was 280 pounds per year (data for annual consumption from Tables 4, 5, 6 7). The annual consumption for each fisherman based on a two year average was lower than that based on a single season because of the aforementioned inconsistency in fishing success from year to year. Although the reported size and species of fish being consumed indicates a significantly contaminated raw food source is being obtained, the average Michigan fisherman apparently does not consume high quantities of fish. The study did confirm that these people eat more fish than the reported national per capita average of 11 pounds per year and that some individuals consume significant amounts of contaminated fish. . PCB's were found in 500 blood and one breast milk specimens collected from participants during the study. The values ranged from a low of 0.007 ppm in the control group to i high of 0.386 ppm in the exposed group. A baseline sample for 1973 (November) and for 1974 (July), a spring sample in 1974, and a series of samples commencing in 1974 and ending in 1975 represent the planned collection schedule for the stulv. The annual fish consumption and the concentrations of I'CB detected in blood collected from participants at the specified times during the study are shown in Tables 4 (Traverse City), 15 HONS 056466 30 Number o f P a rtic ip a n ts Pounds of Lake Michigan Fish Consumed Per Year FIGURE 2. FREQUENCY DISTRIBUTION OF REPORTED GREAT LAKES FISH CONSUMPTION, EXPOSED GROUP, TRAVERSE CITY, MANISTEE C LUD1NGTON 1G HONS 056469 S (Manistee), 6 (ludington), and 7 (South Haven and Algonac). The participants are listed in these tables by ascending order of annual fish consumption as reported for the 1974 baseline year. The raw laboratory data for all PCB tests run are recorded in Appendices A and B. The 35 test results shown in Appendix B are for specimens which were depleted during the development of the final analytical test procedure. These results have been segregated and are not presented in Tables 4,5,667 because it was not possible to retest these specimens with the more precise procedure which was adopted for use in the study. There was a highly significant correlation between the reported quantity of Lake Michigan fish consumed and the concentration of PCB in the blood of participants in the study. Examination of the blood level and fish consumption data for persons from four communities bordering Lake Michigan (Tables 4, 5, 6 6 7) revealed 117 participants who reported eating fish from the lake in 1973. A plot of the levels of PCB in blood vg pounds of fish consumed for this group (Figure 3) indicates a wide range of blood values for each quantity consumed. Using a natural log transformation* of PCB values the data was again plotted (Figure 4) and the correlation between these variables determined to be 0.51 which is highly significant (t = 6.24, p<<.001). Thus higher reported fish consumption was associatec. with more PCB. In addition, a regression analysis of other variables (from Appendix C) showed that being older,and male (height and weight correlated significantly with PCB and with lining male) was significantly associated with higher PCB levels (pc-: 001). The direct relationship between the concentration of PCB in blood and increasing quantities of Lake Michigan fish consumed was observed in all four of the study communities *A log transformation is used when variance increases as values increase (10). Statistical computations were provided by the Office of Vital and Health Statistics, Michigan Department of Public Health. 1? HONS 056<i70 TABLE M. CORRELATION OF FISH EXPOSURE WITH PCB VALUES IN'BLOOD OVER A TWO YEAR PERIOD. TRAVERSE CITY GROUP. / l.D.f 1356 9365ft 0)li0>' Dl'il* .0316' 10316 1318 0319 .0320 . 1368 -032 1 >< -0366ft 5031(2 -03N3 0329ft ;03IS .-03|(6 ;03<(7 0317 0313 0333 0332 ."i)366 J37I 0335 0380 10368ft 0369ft 0327'' :0355 0308ft 0334 0310 037N '337 .'306ft 0363 1353 5392 ' 363 .'.<26 .'301 369 ' i06'ft 122 '370 :369 M09 MO? 16 1973 Fish Consumed 0 lbs. 0 lbs. 0 lbs. 0 lbs. 0 lbs. 0 lbs. 0 lbs. 0 lbs. 0 lbs. D lbs. 1 lbs. 1 lbs. 1 lbs. 0 lbs. 2 lbs. 2 lbs. 0 lbs. 2 lbs. !< lbs. lbs. 28 lbs. 32 lbs. 29 lbs. 65 lbs. 18 lbs. 17 lbs. 17 lbs. 18 lbs. 37 lbs. 22 lbs. 60 lbs. 17 lbs. 35 lbs. 29 lbs. 25 lbs. 27 lbs. 27 lbs. 60 lbs. 32 lbs. 57 lbs. 33 lbs. 3'i lbs. 67 lbs. 56 lbs. 33 lbs. 51 lbs. 29 lbs. 1976 Fish Consumed 0 lbs. 0 lbs. 0 lbs. 0 lbs. 0 lbs. 0 lbs. 1 lbs. 1 lbs. 2 lbs. 2 lbs. 2 lbs. 3 lbs. 5 lbs. 8 lbs. 8 lbs. 9 lbs. 12 lbs. 16 lbs. 16 lbs. 18 lbs. 22 lbs. 22 lbs. 22 lbs. 26 lbs. 25 lbs. 30 lbs. 30 lbs. 30 lbs. 31 lbs. 32 lbs. 33 lbs. 36 lbs. 36 lbs. 39 lbs. 60 lbs. 61 lbs. 61 lbs. Nov.'73 0.033 0.019 0.061 0.027 0.0)1 0.016 0.010 0.013 0.016 0.032 0.015 0.018 0.016 0.022 0.039 0.017 0.015 0.026 0.007 {(ft in': 0.063 0.065 0.020 0.022 0.060 0.020 0.061 0.028 0.060 0.068 0.093 0.075 0.038 is in't 0.055 0.038 0.005 0.037 BLOOD SPECIMENS - Total PCD in parts per mi 11 i on Apr.' 7J1 Jul.176 Jon.'75 Feb.'75 8P-lu25_ May 1 7 0.027 ft* ft* 0.018 0.019 0.011 0.012 0.015 0.015 0.023 0.022 0.019 0.061 0.019 0.022 0.029 0.010 i: it . frit 0.023 0.025 0.061 0.023 0.038 0.051 0.021 0.036 0.016 0.025 0.056 0.032 0.060 0.019 0.026 0.066 0.03! 0.061 0.037 0.077 0.061 0.036 0.016 0.028 ft It 0.033 lV-,Y 0.063 0.053 0.023 0.058 0.061 0.061 0.063 0.103 0.015 0.065 it ft it 0.036 it it 0.0S0 0.036 0.033 0.068 itit 0.037 0.052 0.026 0.053 0.076 0.090 0.012 0.058 0.033 0.036 0.013 0.077 0.036 0.058 0.056 0.066 0.061 0.060 0.039 0.070 0.097 0.012 0.052 0.026 0.036 0.019 0.061 0.090 0.053 0.063 0.062 0.032 0.056 0.086 0.013 0.076 0.013 0.066 0.063 0.029 0.052 0.021 0.033 0.035 0.060 0.066 0. 130 0.013 0.065 0.02 'J 0.03? 0.069 0. OVi 0.035 Nil win p . . 15 HONS 056471 (TABLE 11 Continued) LM >326 -.362 1372 1312 1300* >303' .?304* 1361 0328 1330 1351 0323 0)58 0331 0357 1)11 0356 1350 1373 3367 336*1 1339 1338 1973 F1 sh Consumed 1974 Consumed 32 lbs. i8 lbs. I>8 lbs. SI lbs. 51 lbs. 51 lbs. 53 lbs. 62 lbs. 68 lbs. 2k lbs. 67 lbs. 162 lbs. 260 lbs. 130 lbs. 75 lbs. 93 lbs. 90 lbs. i62 lbs. 60 lbs. 85 lbs. 158 lbs. M lbs. kk lbs. k5 lbs. 47 lbs. 53 lbs. 53 lbs. 56 lbs. 60 lbs. 68 lbs. 79 lbs. 90 lbs. 98 lbs. Ill lbs. 120 lbs. 120 lbs. 131 lbs. 140 lbs. 147 lbs. 152 lbs. 162 lbs. Nov.'73 0.044 0.065 0.060 it if 0.057 0.047 0.030 0.081 0.050 0.152 0.096 It ft 0.083 0.114 BL000 SPECIMENS " Total 1PCB in Darts Apr.'7k Jul'74 Jan.'75 Feb.'75 Apr.'75 0.034 ft* 0.059 0.034 0.045 0.064 *ft ft it O.O69 0.031 0.062 0.043 0.057 0.036 0.047 0.049 0.027 0.045 0.043 0.037 0.044 0.142 0.115 0.022 0.083 0.071 0.007 *** 0.065 0.048 0.042 ft* 0.058 0.131 0.134 0.026 0.068 0.064 ft* 0.064 0.093 0.011 0.072 0.120 0.040 0.058 0.040 0.080 0.078 0.177 0.101 0.021 0.067 0.106 0.076 0.123 0.086 0.078 0.042 0.056 0.045 0.152 0.140 0.073 0.059 0.088 0.081 0.037 0.036 0.135 0.121 0.059 0.064 0.095 May '75 0.07k 0.032 0.061 0.042 0.045 0.092 0.071 0.147 0.172 0.028 0.068 0.121 0.068 0.086 0.014 0.082 0.083 > Chronological order of fish consumed is based on 1973 data. The 1974 questionnaire was not completed. '* Sample was depleted during Initial analysts, using decachloroblphenyl techniques, See Appendix (b) for decachloroblphenyl blood values. Sample lost (broken, not received, etc.) ' 19 HONS 056472 TABLE 6. CORRELATION OF FISH EXPOSURE WITH ,'CB VALUES IN BLOOD OVER A TWO YEAR PERIOD, LUDINGTON GROUP. I.O.f .'020) >0200* ;o206 10207* 30200 20201 70209 70202 .'0205 10208* 70219* 70220* 70221>' '.0212* 70218 7021k 70225* 70215 7022/* 70230 10217 7022k* 70222* 70223* 70229 70216* 70228 70231 70210 0021 1 ,'0213 i'0226 10232 1973 r i sn Consumed 0 lbs. 0 lbs. 1 lbs. 1 lbs. 3 lbs. 3 lbs. 2 lbs. 3 lbs. 2 lbs. 6 lbs. 8 lbs. 8 lbs. 8 lbs. 9 lbs. 22 lbs. 31 lbs. Ik lbs. 26 lbs. 18 lbs. 18 lbs. 29 lbs. 19 lbs. 26 lbs. 26 Iks. 29 lbs. 27 lbs. 27 lbs. 28 lbs. 2k lbs. 2k lbs. 205 lbs. 205 lbs. 197k Consumed 0 lbs. 0 lbs. 1 lbs. 1 lbs. 1 lbs. 2 lbs. k lbs. 10 lbs. 12 lbs. 15 lbs. 18 lbs. 19 lbs. 26 lbs. 27 lbs. 28 lbs. 33 lbs. 39 lbs. 60 lbs. 60 lbs. 60 lbs. Nov.173 0.011 0.017 0.021 0.018 0.037 0.029 0.016 0.015 0.020 0.030 0.0k6 0.016 0.023 0.026 0.023 0.089 0.05k 0.062 0.079 0.060 0.118 0.063 0.036 0.031 0.065 0.065 0.100 0.0k5 BLOOD SPECIMENS " Total PCB In parts per mill Jul. '7k Dec.'7k Jan.'75 Mar.'75 ^ELlLH 0.01k 0.021 0.0k2 0.031 0.016 0.018 0.029 0.022 0.066 0.078 0.055 0.071 0.026 0.069 0.063 0.0k8 0.029 0.059 0.0k7 0.026 0.06k 0.052 0.021 0.031 0.031 0.076 0.077 0.109 0.0k5 0.050 0.019 0.033 0.026 0.085 *** 0.08k 0.035 0.053 0.021 0.022 0.025 0.070 0.066 0.090 0.079 0.017 0.017 0.053 0.070 0.066 0.030 0.021 0.021 0. Ok 6 0.08k 0.065 0.069 0.023 0.055 0.06k 0.023 rti'r/t 0.02 5 0.03 k 0.065 0.066 0.053 * Chronological order of fish consumed Is based on 1973 data. The 197*1 quest lonna i re . j-* not completed. Semple was depleted during Initial analysis, using deeacblorobiphenyl techniques. Appendix (B) for decachloroblphenyl blood values. ** Sample lost (broken, not received, etc.) 20 MONS 056473 TA3LE 5 . CORRELATION OF FISH EXPOSURE WITH PCS VALUES IN ?! COO OVER A TWO YEAR PERIOD, MANISTEE GROUP 10252 -225 i 10256 12230 12233 12255 >0254 0262 0204 -0209 10267 0273 0271 '2208 102 OC* 025! i0270e 0253 0266 0257 0272 2265 0271* 0263 0253 1973 Fish tonsured 0 Tbs. 0 Tbs. 5 Tbs. <6 T bs. <6 lbs. ] b*. 5 lbs. 17 1 bs. 24 lbs. 60 lbs. 22 lbs. 55 Tbs. 26 lbs. 32 lbs. 29 Tbs. 53 lbs. 27 lbs. 30 Tbs. 35 lbs. 48 lbs. 48 1 bs. 79 lbs. 66 lbs. 137i Fish Consumed 0 lbs. 0 Tbs. i Tbs. 1 lbs. 2 Tbs. 2 > UJ. 2 Tbs. 13 Tbs. 15 lbs. 23 lbs. 24 Tbs. 24 lbs. 24 Tbs. 25 1 bs. 28 Tbs. 29 lbs. 30 1 bs. 32 lbs. 35 lbs. 45 Tbs. 48 Tbs. 51 lbs. 64 lbs. Nov-'73 0.014 0.011 0.028 0.039 0.025 0.053 0.027 0.047 0.037 0.025 0.090 0.062 0.094 0.050 0.038 0.249 0.124 0.105 0.151 0.247 0.128 Jol'74 0.035 O.OIO 0.026 0.041 0.029 0.041 0.024 0.023 O.O83 0.022 0.019 0.064 0.117 0.259 0.110 0.096 0.089 0.123 0.239 0.167 0.169 BLOOD SPECIMENS ~ Total PC3 in parts per pillion 10-2-74 10-22-76 Nov.'76 Dec.'76 Jan.'75 Feb.'75 Mar.'75 *a ** 0.069 0.062 0.133 0.019 ** 0.128 0.153 0.020 ** . ** 0.166 0.048 0.025 0.023 0.082 0.027 0.070 0.072 0.097 0.127 0.165 0.206 ** 0.15! 0.057 0.036 0.020 0.091 0.070 0.074 0.227 0.144 0.145 0.185 0.060 07024 0.020 0.042 0.02T 0.018 0.03*> 0.077 0.059 0.331 0.138 0.123 0.099 0.125 0.133 0.131 0.056 0.063 0.244 0.128 0.133 0.180 0.218 . 0.173 0.047 0.035 0.020 0.093 0.C26 0.016 0.070 0.089 0.264 0.102 0.085 0.127 0.229 0.162 0.145 r- Chronological order of fish consumed- is based on 1973 data. The 197** questionnaire was not completed. Sample was depleted during initial analysis, using decachlorobiphenyl techniques. See Appendix (8) for dccachlorobiphenyl blood values. 1- Sample lost (broken, not received, etc.) HONS 056474 tmJLt / . eemr.LLiM t u.i ur Man iami-.i.il in Ill I SOUTH HAVEH AND ALGONAC GROUPS. 1.0. if i00?t 0072 70073 70065 70066 00106 70121 70105 "0115 70070 70126 70067 70111" 70)30 70036 JOI24 :i0119* 70053 70139'' 70140* J0120 .70055 70035 7006 4 10104* 0114 70122 .)0100* 70128 0062 :3116a SOUTH HAVEN______________ 1372 1973 1373 Blood Fish Consumed Fish Specimens Consumed Total PCB(ppm) 28 lbs. 38 lbs. 26 lbs. 51 lbs. 31 lbs. 35 lbs. 26 lbs. 70 lbs. 23 lbs. 26 lbs. 42 lbs. 68 lbs. 2$ lbs. 30 lbs. 26 lbs. 50 lbs. 30 lbs. 50 lbs. 36 lbs. 36 lbs. 24 lbs. 32 lbs. 26 lbs. 43 lbs. 48 lbs. 45 lbs. 60 lbs. 63 lbs. 55 lbs. ' 38 lbs. 80 lbs. <3 lbs. <6 lbs. <10 lbs. 13 lbs. 17 lbs. 17 lbs. 24 lbs. 24 lbs. 26 lbs. 26 lbs. 26 lbs. 26 lbs. 26 lbs. 28 lbs. 28 lbs. 30 lbs. 39 lbs. 40 lbs. 42 lbs. 45 lbs. 55 lbs. 58 -lbs. 64 lbs. 69 lbs. - 0.027 0.029 0.053 0.026 0.035 0.031 0.064 0.045 0.025 0.068 0.059 0.037 0.040 0.034 0.025 0.056 0.164 0.034 0.032 0.018 0.048 0.326 0.043 0.123 0.056 0.029 0.150 0.035 0.082 0.366 0.118 . __ __________ ALGONAC I.D.it' 1972 Fish Consumed 1973 1973 Blood Fish Specimens Consumed Total PCB(p| 10029 11029 10166 10033 10130 10013 10111 10171 10172 10129 10118 10100 10035 10150 10085* 10110 10138 10084* 10168 26 lbs. 26 lbs. 13 lbs. 26 lbs. 31 lbs. 26 lbs. 52 lbs. 39 lbs. 52 lbs. 36 lbs. 26 lbs. 48 lbs. 45 lbs. 77 lbs. 126 lbs. 11 lbs. 11 lbs. 13 lbs. 16 lbs. 20 lbs. 24 lbs. 26 lbs. >26 lbs. >26 lbs. 28 lbs. 34 lbs. 38 lbs. 42 lbs. 26 lbs. 57 lbs. 52 lbs. 135 lbs. 0.023 0.026 0.025 0.030 0.019 o.on 0.021 0.017 0.033 0.030 0.021 0.038 0.012 0.013 0.017 0.020 0.032 0.042 0.026 * Chronological order of f: sh consumed Is based on 1977 data. The 1973 questionnaire wai not completed. 22 HONS 056475 ?CB V a lu e (p a rts p e r m illio n ) Pounds of Fish Consumed Per Year FIGURE 3 CORRELATION OF FCB LEVELS IN HUMAN BLOOD WITH FISH CONSUMPTION 23 KONS 056476 Pounds o f F is h Consumed P e r Y ear 102 82 U 3 9 9 6 62 M M2 22 2r 1.95 e o o 09 0 '9 o c 99 09 9 i9 9 0 9 0 e O 0* 9 <30* o o 6O C9 09 9 6 9 0 C 9 O 9 o 0(5* o C9 9 _L O- 9 O 09 o 9 9 O 0 2.66 3.38 M .09 M . 80 Natural Log of PSB Values (in parts per billion) 5.52 FIGURE M. SCATTER PLOT OF POUNDS OF FISH CONSUMED vs THE NATURAL LOG OF PCB LEVELS IN HUMAN BLOOD HONS 05647? bordering bake Michigan. The mean level of PCD for participants grouped in the major fish consumption categories is shown for each community in Table 8. These data clearly show that the presence of PCB in human blood increases significantly for persons in each ascending consumption category regardless of geographic location. Apparently any exposure to contaminated fish causes an elevation of PCB blood levels. The control group included persons who ate no fish and those who ate up to six pounds per year. A comparison of control participants who ate no fish from Lake Michigan with those who ate an occasional fish revealed significantly higher (p<<,002) PCB levels for the latter grcup (Table 9). It has been reported that the general population is exposed to PCB from a variety of sources and this is reflected in detectable levels in blood (17) and adipose (3, 9). The study confirms such a conclusion as shown by the values'found in the control subjects. It appears that an additional exposure through consumption of contaminated fish, even in small amounts, contributes to the existing body burden of PCB in humans. A substantial exposure to contaminated fish makes a highly significant contribution to the PCB body burden. A comparison of PCB blood values for exposed persons (those who consume 24 or more pounds of Lake Michigan fish per year) in the four communities bordering the lake were compared to the blood values for the zero consumption controls. Using the 1973 baseline data from Tables 4,5,667 the mean PCB values for these two groups were found to be 0.077 ppm (n = 71, range 0.025 to 0.366 ppm) and 0.017 ppm (n = 16, range 0.007 to 0.042 ppm) respectively. The difference between these mean values is highly significant (p<<.001)* indicating a positive correlation between exposure to contam inated fish and the accumulation of PCB in the human body. '''A Cochran adjustment to the t-test was used because of the large difference in the variances of the PCB values for the two groups (18). 25 MONS 056478 The significantly higher PCB levels found in humans is the direct result of an additional exposure to PCB from eating contaminated fish from Lake Michigan. Participants from Algonac, a community bordering Lake St. Clair on the opposite side of the State, who consumed fish from Lake St. Clair, had a mean PCB blood level of 0.023 ppm (n=14, range 0.011 to 0.ON2 ppm)(1973 data from Tables 4, 5, 6 S 7). The difference between the mean PCB values for these two exposed groups is highly significant (p<<.001).* The mean PCB value for the Algonac fish eaters compares closely with that of the Lake Michigan control group (Table 8). Pish in Lake St. Clair do not contain the high levels of PCB reported for Lake Michigan sport fish. Thus, fish eating per se does not cause the observed elevation of PCB in humans but consumption of the more highly contaminated spepies in Lake Michigan does. No annual variation in PCB levels in humans could be demonstrated. The mean PCB values for the control and exposed groups did not appear to change markedly from year to year (1973 baseline v8 1974 baseline, Table 8). The exposed group included individuals reporting many combinations of size, species (contamination level) and quantities of fish consumed. Considering these individual variations in PCB exposure the similarity of the annual mean values strongly suggest that PCB levels in exposed persons as a group do not change significantly from year to year. The PCB blood values for a selection of participants who maintained their usual fish consumption habits during the study period are individually illustrated in figures 10 and 11 (Appendix D). ftA Cochran adjustment of the t-test was used because of the largo difference in the variances of the PCB values for the two groups (18). 26 HONS 056479 TABLE 8 . YEAR TO YEAR COMPARISON OF MEAN PCB VALUES BY CITY City Mean Total PCB Value (ppm) 1973 Participant Group Control Inter mediate Exposed Mean Total PCB Value (ppm) 1974 Participant Group Inter Control mediate Exposed Traverse City 0.017 0.026 0.060 0.019 0.036 0.061 Manistee 0.025 0.068 0.109 C.028 0.029 0.120 Ludington 0.021 0.048 0.062 0.024 0.058 0.055 South Haven 0.028 0.036 0.083 Algonac - 0.024 0.023 Group Mean First 3 Cities 0.020 0.044 0.073 0.023 0.041 0.075 27 HONS 056480 TABLE 9 . EVALUATION OF MEAN PCB VALUES, CONTROL GROUP FISH CONSUMPTION MEAN PCB VALUE (ppm) 1973 MEAN PCB VALUE (ppm) 1974 Ate No Fish 0 lbs./year 0.017 16 0.016 . n = 10 Ate Some Fish 1-6 lbs./year 0.024 H CM II G 0.026 n = 16 . ________ 28 KONS 056481 No seasonal variation in the level of PCB in humans was observed. Initially it was believed that exposure from fish consumption would follow the seasonal pattern of summer fishing and winter lay off. Examination of the dietary data collected during the study shows that this was not the case. Instead, as discussed previously, fish are consumed throughout the year with the higher rates during the active fishing period and lower rates during the winter when preserved fish from the previous season are eaten. The absence of a seasonal consumption pattern was reflected by the absence of statistically significant changes in PCB blood levels during the year (Tables 4,5,6 and Appendix D). The April 1974, November 1973, and July 1974 specimen collections were intended to illustrate blood levels just prior to the resumption of fishing, near the end of fishing, and in the middle of a fishing season respectively. The PCB levels determined for these collections were not significantly different. To test this further an artificially created seasonal comparison was established by comparing blood values for a selection of participants who reportedly ate eight more fish meals over the three month period prior to one specimen collection then in a like period prior to another specimen collection. Fifteen subjects met this arbitrary criteria. A paired t-test of the differences in blood PCB for the high and low consumption periods was not significant (t = 0.99). Abstinence from Lake Michigan fish consumption did not significantly change the level of PCB in the.blood of persons in the exposed group. Forty-four participants from Traverse City, Manistee, and Ludington were selected and asked to abstain from consumption of Lake Michigan fish at the conclusion of the 1974 fishing season. Thirty-six persons were able to maintain an abstinence period of 90 days or more. The 1974 annual consumption, length of abstinence, and PCB blood levels at various times during abstinence for these participants are shown in Table 10. Tho length of abstinence ?9 MONS 056462 Tarsiclpaot "A Ivflojcaa 00318 03715 50723 C0778 03710 00211 Naatata* * 00262 03766 CC269 00267 CC263 00261 0023/ 00265 00263 Travaeaa Cts* *<5332 C0333 O-^f.6 V0555 0C 352 OC353 00)25 00)05 03370 00)02 CC326 ;3->2 00)12 03330 C0)6 W331 00335 00339 00)36 TVla 10. Carralatlaa 7CX tarala la Ki Md Icafth of Abatlaanca froa Mah Cawaaptlaa Aaeval flab Coerasp^lao a Akytlaaan Ibj/r*ar Oatra #0*ra --PC8 Caaceocratiw la glaad farm) Kwubar f Daya Aftar Taralneeloa of flab Coaai 60 75 90 IQS 120 133 150 ;o 12.74/3-75 3 15 11-74/3-75 127 s .2-74/3-75 8S 26 11-74/4-75 158 27 12-74/4-75 126 .033 33 12-74/4-75 121 .005 39 12-74/4-75 121 le .059 .021 .026 .332 .01? J370 JJ66 .017 .021 .086 .065 13 12-76/3-75 108 15 10-75/3-75 167 23 10-75/3-75 176 26 12-74/6-75 176 .082 25 10-74/3-75 176 .069 28 12-74/3-75 108 .072 30 12-74/6-75 176 .097 32 10-74/3-75 148 *5 11-76/2-75 87 51 11-74/3-75 124 .070 .059 .138 .020 .09* .070 .063 .128 .123 .918 .093 .056 1-75/5-75 131 8 1-75/5-75 8 li-74/4-75 132 1* 11-74/5-75 160 30 1-73/5-75 138 .090 30 1-75/5-75 138 .074 31 1-75/4-75 90 .058 36 12-74/5-75 153 36 9-74/5-75 249 .013 .019 41 12-76/5-75 153 .036 .030 46 1-75/4-75 90 .069 .057 64 9-74/3-75 278 47 1-75/5-73 139 .062 .067 56 12-76/6-75 104 79 12-76/5-75 153 .060 .177 .065 .152 00 12-74/3-23 104 .101 .160 129 11-74/5-75 152 12-75/3-75 151 162 1-75/5-73 141 .060 .032 .916 .064 .086 .056 .076 .013 .038 .058 .073 .036 .135 .121 .095 t*4 S 976 rapart *1"* <<'&laroblpSaf* l tchlsi>a. 9a Appaodtx <*> far <acacttlara>lp>iaoy 1 vaWaa. *0NS 056463 ranged from 85 days to 278 days. The date of initiation of abstinence was staggered because of the individuality of the end of the fishing season. In the same manner the end of the abstinence period was staggered and determined by the beginning of the 1975 fishing season or the desire of individuals to avoid wasting their store of frozen catch from the previous season. As discussed previously, this portion of the Study represented a change in the normal habits of most of these subjects, a matter which took considerable self restraint. An examination of the blood levels over the abstaining period (Table 10) shows variation but no pronounced decline in PCB for the group. In fact, more subjects showed no change or a rise than showed a decline in PCB level over time. PCB blood values for selected subjects who abstained from eating fish for part of the study period are individually illustrated in Figures 12 and 13 (Appendix E). An analysis of variance for 24 subjects with a beginning blood PCB level of at least 0.040 ppm showed no significant decline in levels over time. A paired t-test using the initial and final abstaining blood values showed an average net increase in PCB and no statistical significance in the recorded change during abstinence from Lake Michigan fi6h. It must be concluded that abstinence from eating contaminated fish did not reduce the PCB blood levels in these subjects. Thus, an evaluation of the half-life of the concentration of PCB's in humans could not be made within an abstinence period of 90 to 278 days. Cooked Lake Michigan fish ccnsumed by study participants contained PCB's but at levels lower than those reported for raw whole fish. Participants were asked to record their fish consumption in dietary logs and save a portion of their fish meals for analysis. Questionnaire and dietary log data were used to estimate the annual fish consumption reported in Tables 4, 5, 6 6 7. This data also 31 HONS 056484 indicated that Lake Trout, Stcelhoad and Salmon (Coho or Chinook) represented the perferred and most frequently consumed species of fish for most of the participants. An exception was found in South Haven where chubs and perch were eaten more frequently. The PCB levels found in cooked fish collected from participants during 1973 and 1979 are shown in Tabic 11. This data has been averaged and arranged by fish category and community in Table 12. These results represent tests on composites of collected samples of Lake Michigan fish eaten by individuals during the study. The PCB level in cooked Lake Trout ranged from 1.03 to 9.67 ppm, in cooked salmon from 0.98 to 5.38 ppm, and in other cooked fish from 0.36 to 2.06 ppm. These levels are decidedly lower than the level of PCB contamination generally reported for raw whole Trout and Salmon (Table 1). This is not unexpected since preparation.(trimming away fatty tissue) and cooking (loss of fat drippings)* have been shown to decrease the concentration of PCB's (19). The results indicate that actual human exposure to PCB is less than that expected from data for raw whole fish. . The calculated quantity of PCB ingested by eating Lake Michigan fish averaged 96.5 mg/year and ranged from 19.17 to 119.31 mg/year for 91 of the exposed group participant (Table 13). PCB ingestion for each individual was determined by proportioning the reported annual fish consumption by frequency of species eaten and the cooked fish PCB levels (Table 11) for these fish. The community average (Table 12) was used for instances where cooked fish determinations were not available for a participant. Because fish consumption was found to vary from year to year the average annual consumption for the two baseline years of the study was used in each case. f'Tests on two samples of cooking oil used for deep fat frying of fish showed 0.89 and 0.76 ppn PCB. 32 HONS 056485 .TABLE )1. PCB LEVELS IN CQOKLD i'lbli CUNoUiti^u ot uuiuu.u PARTICIPANT I.Dj PCB 1254 CONCENTRATION (parts per mill Ion) . LAKE TROUT SALM >N OTH ER* 1973 1974 1973 197*4 1973 1974 00210 00213 00214 0021? 00223 00225 00227 00231 002IJ 00257 00259 00261 00263 00266 00268 00270 00273 00300 00308 00309 003H 00312 00323 00325 00327 00328 00330 00332 00334 00336 00339 00350 00365 00035 00062 00104 00105 00114 00119 00120 00121 00130 4.51 2.83 3.53 1.63 4.00 3.68 3-75 1.03 I.87 2.48 5.13 3.66 3.44 1.58 3.17 2.23 1.63 3.24 5-17 3.65 2.84 1.48 4.38 2.07 1.97 1.69 4.17 2.35 2.43 1.69 1.63 0.48 1.85 3.86 3.01 . 2.96 3.48 1.87 2.99 3.34 4.89 3.18 2.54 2.64 1.39 0.96 3.66 4.63 2.81 0.76 1.46 O.98 1.32 3.18 1.49 0.90 1.05 1.81 1.69 2.84 4.11 0.73 0.73 0.86 0.62 0.56 1.66 1.17 0.36 4.67 2.84 1.04 3.47 3.18 3.28 3.48 1.66 2.31 1.83 2.32 I.85 2.18 1.33 2.99 2.75 1.29 4.63 3-45 5.38 1.56 2.06 1.87 0.97 0.63 1.52 1.98 0.56 5.63 4.01 5.62 1.45 2.72 2.39 0.47 ^Includes: Pike, Whltefls'), Smelt, Chubs, Menominee and Perch 33 MONS 056466 TABLE 1?. COOKED 1'ISH SURVEY, MEAN PCB CONCEifii 10 BY COMMUNITY LOCATION Traverse Clty: Mean No. Samples Manistee! Mean No. Samples Ludlnpton: Mean No. Samples South Haven: Mean ' No. Samples LAKE TROUT PCB 125k (ppm) 1973 197'* l973-7k FISH SPECIES SALMON PCB 125k (ppm) 1973 197k 1973-7k OTHER* PCB 125k PPm) 1973 197k 197 3-7k 2.80 2.22 II 10 2.52 21 2.78 3.kg 37 3.28 10 . I.k3 1.11 68 1.25 Ik 2.85 2. kO 56 2.60 ` 3-7k 2.20 II 3 k 2.86 7 0.98 0.59 22 0.78 k 3-36 k.02 63 3.58 9 2.50 2.65 7k 2.55 II 1.06 0.86 31 0.99 k A. II 6 2.51 6 1.17 2 "'Includes: Pike, Whlteflsh, Snelt, Chubs, Menominee and Perch HOHs 056467 Table 13 shows that PCB ingestion depends not only on the quantity of fish eaten but also on the species and contamination level of the fish which compose the meals. Thus a person consuming a given quantity of fish could receive a varying amount of PCB depending on the species and the size of fish eaten. In general, PCB ingestion corresponded with the quantity of fish consumed. The correlation between PCB ingestion from fish and human PCB blood levels was significant. An annual PCB dose was determined for each participant listed in Table 13 and compared to the corresponding mean (1973-74 baseline) PCB blood levels as illustrated in Figures 5 (Traverse City group) and 6 (Manistee, Ludington and South Haven groups). The correlation coefficients for'these plots are 0.347 and 0.316 respectively, indicating significance at the 5% probability level. The dose-blood level relationship confirms the pounds of fish consumed - blood level relationship shown previously in Figure 3 and Table 8. Interpretation of the meaning of these correlations requires caution because of the number of uncontrolled variables which complicates accurate estimation of PCB ingestion. The PCB level in each fish eaten, the exact number of pounds constmed annually, the species and quantity of fish consumed in previous years, and the conditions controlling the appearance and level of PCB in human blood are: all factors which contribute to the high degree of scatter seen in the data plots. . The maximum allowable ingestion recommended for protracted exposure to PCB is being exceeded by the majority of the exposed group participants. It has been estimated that 1 pg/Kg body weight/day represents the maximum allowable dose for protracted ingestion of PCB's (8). This was based on extrapolation from the lowest total dose (500 mg) reported to produce an effect in man and the assumption that PCB exposure may be tolerated over a long period if daily exposure is hold to minimal levels. The calculited mean daily dose 35 HOAS 056468 TABl-E 13. ESTIMATED PCB INGESTION FROM FISH, ANNUAL DOSE AND MEAN BLOOD VALUES FOR PERSONS CONSUMING 24 OR MORE POUNDS PER YEAR I.D. f Body Weight TRAVERSE CITY: 00306 00363 00337 00329 00355 00353 00352 00339 00325 00335 00369 00336 00365 00370 00309 00326 00351 00301 00372 00362 00312 00369 00300 00304 00322 00361 00328 00330 00323 00358 00356 00364 79 77 97 82 75 64 74 79 64 61 57 77 75 82 100 71 61 ' 70 54 70 59 136 64 57 66 70 64 54 75 73 100 77 1973-75 Mean Annual Fish Consumption Rg/yr. 11.3 11.8 12.2 12.7 12.7 13.2 13.6 14.1 14.5 14.5 15.0 15.9 15.9 16.3 17.0 17.2 19.1 20.4 20.4 20.8 21.8 21.8 23.1 23.1 23.1 24.0 26.3 28.1 30.8 35.8 44.5 47.2 Calculated Annual1'1 PCB Ingestion From Fish mg PCB/yr. Annual PCB Dose mg/Kg/vr. Mean Blood** PCB Value --LPi>J!>_____ 14.17 0.18 23.85 0.30 24.77 0.26 18.65 0.23 22.33 0.28 37.14 0.59 38.43 0.52 36.31t 0.48 33.82+ 0.53 37.48 0.61 30.01 0.53 31.09+ 0.40 23.66+ 0.32 46.11 0.68 28.01+ 0.28 40.16 0.57 26.97 0.44 35.57 0.51 35.57 0.67 69.30 0.99 29.09+ 0.49 38.28 0.28 40.65+ 0.64 40.65 0.71 65.33 0.99 48.37 0.68 78.98+ 1.24 . 45.39+ 0.83 44.65+ 0.60 44.79 0.62 78.15 0.78 94.91 1.23 0.040 0.04 0 0.041 0.050 0.043 0.056 0.098 0.047 0.070 0.042 0.036 0.037 0.015 0.034 0.048 0.052 0.081 0.038 0.045 0.034 0.065 0.033 0.060 0.067 0.068 0.065 0.048 0.036 0.054 0.131 0.064 0.009 * Values for PCB concentrations in fish provided from tests on cooled composite samples provided by participants or from community averages for such samples. t Determined from tests on fish samples provided by the participant AA Average of the November 1973 and July 1979 blood values 36 HONS 056489 TABLE 13. (continued) i.d.i? Body Weight Kg 1973-75 Mean Annual Fish Consumption Kg/yr. TRAVERSE CITY (continued) 00350 00373 00339 00311 00331 00338 84 68 63 102 82 81 48.5 50.8 53.9 57.2 57.2 36.1 Calculated Annual1'1 PCB Ingestion From Fish mg PCB/yr. 7 5.23t 89.22 90.57t 100.47t 92.27 102.80 Annual PCB Dose m^/Kg/yr. 0.90 1.31 1.44 0.99 1.13 1.25 Mean Blood PCB Valu (pom) 0.106 0.064 0.078 0.072 0.143 0.117 MANISTEE 00273 00271 00288 00260 00261 00270 00266 00257 00269 00272 00269 00265 00274 00263 00258 82 69 75 65 ` 91 73 76 66 104 84 94 104 70 106 82 10.9 10.9 11.1 11.8 13.6 13.2 13.2 14.1 14.5 15.9 18.6 21.3 21.8 29.5 24.5 17.53t 29.43 44.53t 23.02 35.68t 45.52t 25.OOt 43.64t 35.62 4 2.93 36.2 9t 74.42 35.06 103.67t 76.03 0.22 0.42 0.60 0.35 6.39 0.63 0.33 0.66 0.34 0.51 0.38 0.71 0.50 0.98 0.93 0.022 0.019 0.062 0.094 0.084 0.038 0.110 0.096 0.023 0.089 0.259 0.137 0.239 0.207 0.148 LUDINGT0N 00217 00223 00229 00216 00228 00231 00210 94 79 57 67 102 64 68 - 10.9 11.8 12.5 12.3 12.3 12.7 12.9 36.88+ 29.65+ 24.02 30.60 23.79 51.34+ 37.18+ . 0.39 0.37 0.42 0.46 0.23 0.81 0.54 . 0.071 0.036 0.021 0.031 0.031 0.031 0.076 Values for PCB concentrations in fish provided from tests on coo .cd composite samples provided by participants or from community averages for such samples. t Determined from tests on fish samples provided by the participant Average of the November 1973 and July 1974 blood values 37 MONS 056490 TABLE 13. (continued) I. D. # Body Weight Ke 1973-75 Mean Annual Fish Consumption Ka/yr. LUDINGTON (continued) 00211 00232 00226 00213 64 93 70 81 14.3 27.2 60.1 60.1 Calculated Annual* PCB Ingestion From Fish ' mg PCB/yr. Annual PCB Dose mg/Kg/yr. 40.99 109.01 49.76 78.29+ 0.64 1.18 0.70 . 0.96 Mean Blood** PCB Value (ppm) 0.077 0.107 0.045 0.050 SOUTH HAVEN 00066 00115 00121 00070 00106 00111 00036 00130 00119 0006S 00120 00035 00126 00055 00140 00139 00124 00063 00064 00067 00105 00104 00114 00122 00120 00116 00062 52 52 61 74 73 91 57 73 84 75 61 75 61 74 68 55 67 89 79 82 95 79 82 101 86 79 84 10.9 11.3 11.3 11.8 11.8 11.8 12.2 12.7 13.6 14.5 14.5 15.4 15.4 16.3 16.3 17.2 17.7 18.1 19.9 21.3 21.3 21.8 22.7 26.8 27.2 36.3 38.1 . 18.56 35.72 41.75+ 29.16 31.87 27.66 29.80 28.48t 42.49t 24.75 57.68+ 37.S3t 26.31 56.66 38.36 40.44 41.50 42.56 46.82 36.37 57.61+ 44.29+ 71.4 5+ 45.66 46.44 114.31 84.58+ 0.36 0.69 0.68 0.40 0.44 0.31 0.53 0.39 0.51 0.33 0.94 0.50 0.43 0.76 0.56 0.73 0.73 0.4 8 0.59 0.46 0.61 0.56 0.88 0.45 0.54 1.44 1.01 ' . 0.035 0.025 0.064 0.068 0.031 0.040 0.025 0.034 0.164 0.026 0.048 0.043 0.059 0.326 0.018 0.032 0.056 0.034 0.123 0.037 0.045 0.056 0.029 0.150 0.082 0.118 0.366 f` Values for PCB concentrations in fish provided from tests on cooked composite samples provided by participants or from community averages for isuch samples. t Determined from tests on fish samples provided by the participant *>' Average of the November 1973 and July 1974 blood values 38 HOMS 056491 PCB C o n c e n tr a tio n - B lo o d (ppm ) FIGURE 5 . CORRELATION OF FOB LEVELS IN HUMAN BLOOD WITH PCB INGESTION FROM FISH, TRAVERSE CITT GROUP 39 KONS 056492 PCB C o n c e n tr a tio n - B lo o d (ppm ) 0.366 0.296 0.227 0.157 0.088 0 S. '3 '*. o 5 o 0 0.018 jo........-j 0.220 0.M6M 0.708 0.952 1.196 Annual PCB Dose from Fish (mg/Kg/yr.) 1. 0 FIGURE 6 . COP.RELATION OF PCB LEVELS IN HUMAN BLOOD WITH PCB INGESTION FROM FISH, MANISTEE, LUDINGTON, AND SOUTH HAVEN GROUPS 9 0 HONS 056493 received by the exposed group listed in Table 13 is 1.70 pg/Kg/day and ranges from 0.49 to 3.94 pg/Kg/day (Table 14). Except for those in the lower range these levels clearly exceed the recommended dose established for long term continuous exposure. Lake Michigan fish consumption has been shown to be intermittently continuous through the year. This provides repeated doses of PCB which appear sufficient to maintain a PCB equilibrium within humans. Sport fishing has become increasingly popular in the past eight years and activity is expected to continue. Consumption of salmonids from the Great Lakes is and will continue to represent a source of prolonged exposure to PCB. If the average annual rate of PCB ingestion from fish shown in Table 14 is continued over the years, and if net accumulation occurs,the average fish eater could receive a total PCB dose equal to the 200 mg safety limit in 4.3 years. The lowest total dose reported to produce an overt effect in humans (S00 mg) would be reached in 10.7 years. An individual who habitually consumed large quantities of contaminated fish would reach these total dose levels sooner, 1.75 years and 4.3 years respectively if the top range value is used. Such a person would reach the average total dose received by those with Yusho symptoms (2,000 mg) in 17.5 years. If it is proven that such total dose accumulations actually occur in persons who consume significant quantities of contaminated Great Lakes fish, then the long term exposure to PCB contaminated fish could prove: important. Segregation of exposed participants on the basis of reported prior history of fish consumption revealed that PCB blood levels rise with the length of tine fish have been consumed (Table 15). All of the groups received comparable PCB doses during 1973-74 yet the mean blood PCB levels for this period were directly related to the extent of prior fish consumption. Higher blood levels were (associated with a longer history of consuming Lake Michigan fish. Although the details and quantities of prior fish consumption are not 41 HONS 056494 TABLE 14. CALCULATED PCB INGESTION AND DOSE FROM CONSUMPTION OF GREAT LAKES FISH* PCB Ingestion Annual mg/year Daily ' pg/day PCB Dose Annual mg/Kg/year Daily pg/Kg/day Mean 46.S00 0.130 Mean 0.620 1.700 Range: low 14.170 high 114.310 0.038 0.313 Range: low 0.180 high 1.440 '`Calculated from data on 91 exposed subjects in Table 13. 0.490 3.940 . .. M2 HONS 056495 TABLE 15. CORRELATION OF BLOOD PCB LEVELS WITH NUMBER OF YEARS OF FISH CONSUMPTION No. Years Lake Michigan Fish Eaten 1-3 4 -6 7-9 10 or more Mean PCB Dose mg/Kg/year 0.58 0.62 0.53 0.59 Mean Blood PCB No. ...... _____________ Subjects* 0.046 0.055 0.068 0.082 3 35 6 43 ''From Traverse City, Manistee, Ludington and South Haven h 3 HONS 056496 known the data strongly suggest that long term exposure to PCB contaminated fish results in a net accumulation of PCB in human tissue. Furthermore, the absence of any data indicating a decline in PCB levels suggests that a PCB equilibrium may exist and that recirculation of PCB may be occurring in humans. This would cause target organs to be continuously exposed to PCB for years. Exposed and control participants were asked if IT n they had experienced any of 17 health conditions. Many of . the conditions described the signs and symptoms reported for Yusho disease. There was no significant difference between the number of exposed and control persons responding "yes" ! to any condition (Table 16). No health conditions or group ' ., i of symptoms could be identified that were clearly related to PCB exposure. This implies that exposure to PCB from eating' contaminated fish and the presence of PCB in exposed persons had not caused an observable acute effect in humans at the time of the study. This does not exclude the possibility that effects too subtle for detection by our survey are occurring. The blood PCB levels for those responding "yes" to a health condition were not significantly different from those responding "no" to the condition (Table 17). Standard t-tests and a Behrens-Fisher technique for unequal variances were used to evaluate the data. A significant number of exposed subjects or subjects with high PCB levels would be expected to report a given health condition if such a condition was a manifestation of the presence of PCB in humans. The failure of this to occur suggests that there was no common illness related to PCB exposure or tha~ PCB blood levels were not related to the appearance of any of the conditions investigated. M'l HONS 056497 TABU: 16. EXPOSED AND CONTROL PARTICIPANT RESPONSES' TO HEALTH CONDITIONS Health Conditions i l i. Stomach pains or vomlttlng for more than one week. 2. Kidney and/or bladder problems. 3. Stroke. k. General Fatigue 5. Persistent headaches. 6. Weakness or loss of movement In any part of body. 7. Difficulty with coordination. .8 Fainting, dizzy spells or loss of consciousness. 9. Numbness or tingling In arms or legs. 10. Loss of memory (for names or numbers). II. Shaking, unsteadiness, especially of hands and arms. 12. Problems with speech or writing. 13. Convulsions or a seizure Ik. Persistent blurred vision or unusual sensitivity to light. 15. Discharge from eyes or swelling of the eyelids. 16, Persistent skin eruptions or unusual skin pigmentation or Itching.' 17. Increased perspiration of palms. 18. Other . Fish Eater Yes No 1 75 k 72 0 76 7 69 5 71 1 75 2 7<t 5 71 5 71 0 76 1 75 0 76 0 76 3 73 1 75 1 75 1 75 7 69 Non-Fish Eater Yes No Fisher's Probab11ity 1 37 2 36 0 38 2 36 1 37 1 37 0 38 1 37 0 38 0 38 0 0' 0 CO 38 38 0 38 1 37 0 38 0 38 . 0 38 0.56 0.68 .. 0.37 0.35 0.56 0.MM 0.13 __ 0.67 --- 0.29 0.56 0.67 " 0.67 0.C5 Any of the above (at least one). 2M 52 7 31 0.10 "Fisher's exact test of the difference between "yes" proportions for the two groups. MS HOHS 056498 I TABLE 17. MEAN AMD STANDARD DEVIATIONS Or PCB VALUES AND SAMPLE SIZES TOR EACH CONDITION BY REPORTING STATUS 1 y * Condition Reporter Having Condition Standard Mean of Jeviatlon of PCB Values PCB Values Sample . ..(ppm).... (ppm) S 1 zc Reported Not Having Condi t Ion Standard Mean of Deviation of PCB Values PCB Values Sample . (ppm) (ppm) Size 1. Stomach 2. Kidney/Bladder J. Stroke 4. Fatigue 5. Headaches | 6. Weakness/Loss of Movement 7- Coordination 8. Falntlng/Dlzzlness 9. Numbness/Tingling 1 10. Lost of Memory II. Shaklng/llnsteadlness 12. Speech/Writing | 1}. Convulsions | IM. Blurred Vision 15. Eye Discharge 1 16. Skin i 17. Balm Perspiration ! 18. Other j 19. At Leatt One of Above ii 0.058 0.036 O.OM2** 0.0M7 0.056 0.0MI 0.036 O.0M7** 0.015** 0.051 --0.088** 0.0M7 0.057 0.028 0.067** 0.051 0.055 0.020 0.026 0.051 0.03M d.0M8 0.029 0.028 0.006 0.036 --0.102 0.02M 0.032 0.009 0.077 0.0M6 7 0.0M9 IM 0.051 -IM 0.051 19 0.050 8 0.0M9 2 0.050 12 0.051 15 0.050 M 0.051 M 0.050 1* 0* -M 0.0M9 6 0.050 8 0.0M9 2 0.050 IM 0.0M8 6M 0.0M9 . * No test possible; only one or no cases. i'(i Unequal variances. These were tested using a standard F-ratlo test. 0.0M3 0.0M5 00 0.0M5 0.0M2 123 116 129 116 111 O.OMM 122 0.0MM 128 0.0M5 118 O.0M5 115 O.OMM 126 O.OMM 126 -- 129 -- 130 O.OMt 126 O.OMM I2M O.OMM 122 O.OMM 128 0.038 116 0.0MI 66 MS WONS 056499 RESIDUE ANALYTICAL METHODOLOGY: The development of a suitable method for detefction and quantitation of PCB's in human blood involved the following investigations: (1) extraction, (2) separation from organochlorine pesticides, (3) derivation, and (4) gas chromatography. Practicality of the analytical scheme, time involved per sample, and reliability of data obtained were important factors in the choice of procedures. Extraction, separation by silica gel chromatography and direct GLC determination of Aroclors (PCB's) requires about half as much laboratory time as the combination of extraction, separation by chemical degradation, perchlorination, and GLC determination of decachlorobiphenyl. At best, either scheme can provide only a good estimate of the degree, of exposure of the individual to the PCB compounds actually present in the sample. Accumulation of PCB's in the biological system appears to be higher in the case of pentachlorobiphenyl and more highly chlorinated biphenyls (20). An extraction procedure for pesticide residue analysis of blood developed specifically for Corrmunity Pesticide Study programs (21) was investigated initially. Burke (22) questioned whether the two-hour extraction with hexane on the rotary mixer is efficient. Trial runs indicated less than 45% of Aroclor 1260 was recovered-from sheep plasma fortified at the 100 ppb level. Some residue analysts had pretreated blood samples with 60% sulfuric acid to release pesticide residues bound to protein sites (23). Our preliminary investigation of this method produced about 55% recovery of Aroclor 1260 from spiked sh ep plasma. The procedure currently used for extracting PCB's from blood samples incorporates a triple solvent system (i.e., methanol, ethyl ether and hexane), is highly reproducible, and is more quantitative. Since PCB's are fat soluble chemicals 47 HONS 056500 and the lipids in blood exist mainly as conjugates with protein (24), a method designed to disrupt lipoprotein complexes was required for efficient extraction of blood lipids and, hence, PCB's. Plasma proteins are denatured with methanol, and lipids are extracted with a mixture of ethyl ether and hexane. Recoveries of PCB's (as Aroclors) greater than 88% have been achieved, and a better match of the profiles for extracted Aroclors with those for reference standards have been observed with this method. Separation of PCB's from organochlorine pesticides is necessary with either direct analysis or derivation in order to reduce or eliminate interference by DDT and its isomers. Although numerous separation techniques have been reported in scientific journals, efforts were focused on charcoal column chromatography, chemical degradation of organochlorine pesticides, and silica gel column chromatography. These are more suited to detecting the small quantities of PCB found in human blood samples than macro methods such as the silicic acid-Celite column chromatography (25). The charcoal column technique described by Berg et al. (26) appeared to meet our requirement for handling micro quantities of PCB's (5 micrograms or less). A 6 mm i.d. Chromaflex column (Kontes) is plugged with glass wool and filled with 1.1 gram Fisher No. 5-690 charcoal as an acetone slurry. An acetone solution of the sample is added on the column. The DDT group pesticides are eluted with 90 ml of Solvent A (25% acetone in ethyl ether) and PCB's with 60 ml of benzene. Under these conditons, some PCB's eluted in the first fraction but increasing the weight of charcoal from 1.1 gram to 2.5 grans reduced this effect. Although the separation of PCB's appeared to be satisfactory (80%), the elution required as much as three hours per sample, probably due to the formation of vapor pockets. A micro scale procedure for the quantitative separat;on of PCB's from DDT and its analogs was studied in a cooperative effort by Dr. Trotter (27). Certain organochlorine pesticides li a HONS 056501 arc dehydrochlorinated with ethanolic potassium hydroxide to their respective olefins (28) which are then oxidized by chromic acid in acetic acid to dichlorobenzophenone (29). The latter is separated from PCB's by Florisil column chromatography. Trotter (30) achieved recoveries greater than 80% in the ppm range (based on a 5-gram sample) and recoveries of 60% or more in the ppb range for Aroclor 1254 and 1260. Chemical degradation has little effect on certain pesticides such as dieldrin, oxychlordane and Mirex which may not 'be at levels sufficient to interfere with the quantitation of PCB's in human blood. Our chemical degradation experiments with 0.5 |ig to 1.0 pg amounts of Arochlor 1254 and 1260 indicated that recoveries of 65-90% can be achieved. In an effort to seek a more simplified and less time consuming technique, attention was turned to the use of a micro column of silica gel (31). Woelm silica gel, activity grade I, deactivated with 3% (v/w) distilled water, was sandwiched between layers of anhydrous sodium sulfate in a 7 mm i.d. Chroriaflex column. This column was tested with standard solutions of Aroclor 1254 (2.0 pg) and organochlorine pesticides by elution with hexane. With the exception of p,p' DDE, pesticides were found to be completely separated from Aroclor 1254 (about 95% recovery). However, the presence of DDE did not interfere with direct analysis of PCB's. Elution of the PCB fraction can be accomplished in 20-25 minutes. Perchlorination of PCB's to decachlorobiphenyl (DCB) with antimony pentachloride (32) was .evaluated for improved sensitivity of PCB analysis. Conversion of a multicomponent residue to a single compound offered several advantages. This procedure would provide a qualitative and quantitative confirmatory method for PCB's. Also, it would minimize the necessary analytical judgments involved in the measurement of gas chromatographic tracings of a multicomponent residue. Considerable effort was devoted to adapting this procedure for 49 NONS 056502 routine analysis of PCB's in human blood samples, and several problems became evident. At less than 50 ppb PCB levels, conversions were less reproducible and reagent blanks became significant, e.g., 2-10 ppb DOB for a 5-gram sample. Antimony pentaohloride has been found to contain a bromide contaminant which produced bromononachlorobiphenyl during perchlorination (33), but several lots of a high purity grade (99.99%) received in this laboratory did not show any evidence of this contaminant. . The suggestion that PCB levels in blood samples can be determined directly (27) appeared to be more practical for this survey because the gas chromatograph response can be adjusted to permit detection levels as low as 10 ppb total PCB's (Aroclor 1254 and 1260) and fewer steps would be involved in preparing samples for gas chromatographic analysis. The GLC column used for the determination of individual Aroclors was 4%SE-30/6%OV-210. Estimation of PCB residue levels in sample extracts was based on the heights of five peaks in comparison with respective peaks derived from reference Aroclors. The GLC peak having a relative retention time of 1.26 (p,p' DDE=1.00; T-203oC) cn this column was distinct and was selected for the estimation of Aroclor 1254 levels. The analytical procedure incorporating extraction, separation by silica gel chromatography, and direct determination by gas chromatography was the final method of choice primarily because of its superior analytical proc.uctivity rate. The output by this procedure was approximately 4 samples per man-day, whereas only 2 samples per man-day could be processed according to the analytical scheme comprising extraction, spearation by chemical degradation, perchlorination, and GLC determination of decachlorobiphenyl. The analyst had less confidence in the latter procedure as yields of the derivative were variable, particularly at the low I'CB levels often found 50 HONS 056503 in blood samples. Also at low residue levels, reagent blanks contributed a significant proportion of the decachlorobiphenyl product and were not always dependable. CHOSEN ANALYTICAL PROCEDURE: I. Reagents: A. Solvents: Hexane, ethyl ether and methanol suitable for use in pesticide residue analysis or distilled-inglass (Burdick and Jackson Laboratories, Inc. 1953 S. Harvey St., Muskegon, Mi. 99442). B. Silica gel: Woelm, activity grade I (ICN Pharmaceutical Inc.). Deactivate with 3% (v/w) distilled water and store in tightly sealed glass container. C. Sodium sulfate: Anhydrous, granular (Mallinckrodt No. 8024 or equivalent). Store at 130 in glass stoppered bottle. '' D. Antimony pentachloride: Anhydrous, 99.99% (Research Organic./lnorganic Chemical Corp., No. SB-03). E. Reference standards: Aroclor 1254 (FDA 370) and decachlorobiphenyl (FDA 1056) obtained from Division of Chemical Technology, Bureau of Foods, FDA, Washington D.C.; Aroclor 1260 from P.T.S.E.L., U.S.E.P.A., Research Triagle Park, N.C. II. Apparatus: A. Chromatographic column; Chromaflex glass column, 7 mm i.d. (Kontes No. K-420100-0022). Prepare columns by plugging with small wad of clean glass wool, filling to depth of 1 cm with granular anhydrous NajSOi,, adding 3.0 grams absorbent, and topping with another 2-3 cm layer of NarSOi,. 51 HONS 056504 B. Concentrating apparatus: Evaporative concentrator complete with modified micro Snyder column, joint 19/22 (Kontes No. K-S69250); concentrator tubes sizes 1025 and 2525 (Kontes No. K-570050). C. Gas chromatograph: Tracor Model MT220 with tritium foil parallel-plate design electron capture detectors; glass, U-shaped columns, S' x 4 mm i.d. 1. Pack first column with 4%SE-30/6?.OV-210 on 80-100 mesh Gas Chrom Q. 2. Pack second column with 54CV-210 on 80-100 mesh Supelcoport. 3. Pack third column with 1WV-101 on 100-120 mesh Gas Chrom Q. 4. Columns 1 and 2 are used for direct analysis of PCB's and Column 3 for decachlorobiphenyl. 5. Operating conditions: Nitrogen flow, 100 ml/min. (Columns 1 and 3) and 40 ml/min. (Column 2); column and detector temperatures, 203C; injector temperature, 225C. III. Extraction: Transfer 5.0 ml plasma or serum into clean culture tube (20 x 150 mm) having Teflon-lined screw cap. Add 4 ml methanol and nwirl on Vortex mixer about 10 seconds. Add 5 ml hexane-ethyl ether (1:1) and attach screw cap securely. Mix contents 10 minutes, using rotary mixer at 50-55 rpm. Centrifuge for 2-5 minutes at 2000 rpm. Transfer upper layer to 25 ml concentrator tube. Repeat extraction twice with 5 ml hexane-ethyl ether. Add carborundum chip, attach micro Snycier column and concentrate extract to about 0.5 ml using a water bath at 80-90C. 52 HONS 056505 IV. Separation on Silica Gel: VJash silica gel chromatographic column with 5 ml hexane. Just as the last of the hexane enters sodium sulfate layer, quantitatively transfer concentrated extract to the top of the column using a disposable glass pipet. Elute PCB's from column with 19 ml hexane. Collect a first fraction, 4 ml, and a second fraction, 15 ml, in graduated centrifuge tubes. Discard the first fraction. - The second fraction contains PCB's and p,p' DDE. Reduce volume to 1.0 ml with a slight stream of nitrogen directed upon hexane solution. If transferred to a 10-ml concentrator tube, the extract may be reduced to 0.5 ml. V. Gas Chromatographic Analysis: Inject a 5-8 pi aliquot of concentrated eluate, and, if necessary, adjust to' suitable volume. Heights of peaks having the following relative retention times (p,p DDE=1.00) are measured and compared with respective peaks for reference standards of Aroclor 1254 and 1260: 1.26; 1.42; 1.69; 1.85 and 2.62 (4%SE-30/6%OV-210 column at 203C). Aroclor 1251 is calculated on the basis of peak 1.26. Peak 1.86 is considered to be derived from Aroclor 1260. After subtracting apparent Aroclor 1259 portions, i.e., peak heights, of peaks 1.42, 1.69 and 2.62, the mean Aroclor 1260 level of these peaks is then calculated. No corrections were made to reflect 100% recovery of PCB's. The limit of detection is 10 ppb Aroclor 1254 and 6 ppb Aroclor 1260. VI. Controls and Reagent Blanks: Each batch of 12 samples consists cf one control and one reagent blank. Quality control samples are prepared by fortifying sheep plasma with ethyl acetate solutions of Aroclor 1254 (66-132 ppb) and Aroclor 1260 (16-30 ppb). Recoveries were 91% Aroclor 1254 (range 80-105%) and 85% Aroclor 1260 (range 75-96%). PCB's were not detected in sheep plasma blanks used in this study. S3 HONS 056506 VII. Confirmation by Perchlorination: The procedure by Armour (32) was followed for qualitative confirmation of PCB's in certain blood samples. VIII. Precision: Sixty-seven blood samples were analyzed in duplicates by the direct analysis procedure. The total levels for the first reading were correlated with those of the second reading as a test of reliability (Figure 7). The correlation coefficient was 0.972 which indicates a highly significant agreement for the replicate tests and a high degree of reliability of analytical data obtained in this study. SPECIAL TESTS: I. Figures B and 9 are illustrations of the chromatograms of hexane extracts following chemical degradation of samples containing Aroclor 1254 and 1260 respectively. Plot B in Figure 8 represents a sample of cooking oil used by one of the study subjects for fish preparation. As 6hown the PCB found in the cooking oil closely resembles the GLC tracing for the Aroclor 1254 standard (plot A . Figure 8). The absence of pronounced peaks indicative of Aroclor 1260 in the oil used to cook Lake Michigan fish and the Aroclor concentrations found in the blood of participants (Appendix A) jndica:e that those who consume Lake Michigan fish are primarily exposed to the Aroclor 1254 form of PCB. II. Best Specimen: Fasting and nonfasting blood specimens were collected from subjects in the PCB Study and an investigation involving a related chemical, polybrominated biphenyl (PBB). There 54 MONS 05650? was no significant difference in the chlorinated or brominated biphenyl concentrations for the two types of blood specimens (Table 18). Thus either type of sample would be satisfactory for the detection of PCS. III. Breast Milk Specimen: Traverse City participant 00324 who consumed an average of 28 pounds of Lake Michigan fish during 1973-74 gave birth to a child in January of 1975. A breast milk specimen collected during January contained 4 ppm PCD, , Aroclor 1254 (fat basis - the specimen had 2% fat content). In contrast, a blood specimen collected in January contained 0.053 ppm total PCB. This shows that body tissues with a significant lipid content will contain . PCB's which are fat soluble at levels greater than those found in blood. It also suggests that pregnant or nursing women should consume Lake Michigan fish with caution. 55 WOKS 056508 24 5 T o t a l PCB (p p b ) Run # 1 . 198 151 104 57 & *#*> oo CJ> o e 10 10 _u 57 104 1S1 Total PCB (ppb) Run #2 198 FIGURE 7 . ANALYTICAL PRECISION, PLOT OF PCB VALUES FOR REPLICATE TESTS. 245 56 HONS 056509 I Fig.O.A, 1. Sng Aroclor 1254. B, 1. 34mg cooking oil (00268). GLC parameters: see Fig. 9. MONS 056510 24 min I II II I Fig. 9. Aroclor 1260 (0. 6 ng). GLC parameters, --column: 6'x4 5% OV-210; col. T, : 168C. Arrow: p.p' DDT elution position. Flow: 70ml N.j/mln. $8 HONS 056511 TABLE 18. BEST SPECIMEN COMPARISON HALOGENATED BIPHENYL VALUE (ppm) Fasting Blood Nonfasting Blood 0.01S O.OS1 0.001 0.001 0.066 0.03S 0.088 0.005 0.022 0.042 0.025 0.017 0.050 0.001 0.001 0.065 0.038 0.102 0.005 0.025 0.040 0.025 HONS 056512 59 APPENDIX A FINAL LABORATORY DATA FOR PCB TESTS ON BLOOD SPECIMENS COLLECTED DURING THE STUDY 60 HONS 056513 parts per million Collection PCB PCB PCB IDA Date 1256 1260 Total parts per nit 11 i or. Col lection PCB PC8 PCB ID# Date 1256 1260 Toto 1 00035 00035 00053 00055 00062 0006l| 00065 00066 00067 00070 00071 00072 00073 00100 00106 00105 00106 00111 00116 00H5 00115 00119 00120 00121 00122 00126 00126 00128 00130 00139 00160 00200 00200 00201 00201 00202 00202 00203 00203 00206 00205 00205 00206 00206 00207 00208 00:09 00209 00210 00210 29-10-73 29-10-73 29-10-73 29-10-73 27-10-73 27-10-73 29-10-73 29-10-73 29- 10-73 29-10-73 27-10-73 29-10-73 29-10-73 29-10-73 29-10-73 29-10-73 29-10-73 29-10-73 29-10-73 29-10-73 29-10-73 29-10-73 29-10-73 29-10-73 29-10-73 29-10-73 29-10-73 29-10-73 29-10-73 29-10-73 29-10-73 01-11-73 11-07-76 01-11-73 11-07-76 01-11-73 29-07-76 01-11-73 11-07-76 01-11-73 01-11-73 11-07-76 01-11-73 11-07-76 01-11-73 01-11-73 01-11-73 29-07-76 01-11-73 11-07-76 0.030 0.020 0.026 0.218 0.255 0.091 0.018 0.027 0.027 0.065 0.022 0.022 0.038 0.027 0.062 0.032 0.025 0.033 0.019 0.018 0.075 0.126 0.060 0.069 0.116 0.036 0.068 0.066 0.025 0.023 0.012 0.027 0.033 0.023 0.026 0.012 0.015 0.009 0.011 0.015 0.015 0.022 0.016 0.016 0.016 0.022 0.012 0.012 0.068 0.057 0.013 0.005 0.010 0.108 0.111 0.032 0.008 0.008 0.010 0.023 0.005 0.007 0.015 0.008 0.016 0.013 0.006 0.007 0.010 0.007 0.063 0.038 0.008 0.015 0.036 0.020 0.011 0.018 0.009 0.009 0.006 0.010 0.009 0.006 0.005 0.003 0.003 0.002 0.003 0.002 0.005 0.007 0.005 0.005 0.006 0.008 0.006 0.006 0.017 0.019 0.063 0.025 0.036 0.326 0.366 0.123 0.026 0.035 0.037 0.068 0.027 0.029 0.053 0.035 0.056 0.065 0.031 0.060 0.029 0.025 0.118 0.166 0.068 0.066 0.150 0.056 0.059 0.082 0.036 0.032 0.018 0.037 0.062 0.029 0.031 0.015 0.018 0.011 0.016 0.017 0.020 0.029 0.021 0.021 0.018 0.030 0.016 0.016 0.065 0.076 . 00210 00210 00210 00210 00210 00211 00211 00211 00211 0021 1 0021 1 00212 00213 00213 00213 00213 00213 00216 00216 00216 00215 00215 00215 00215 00215 00215 00216 0021V 0021'/ 0021V 0021V 0021V 00217 002111 00218 00216 00218 00216 .00219 00219 00220 0022 i 0022;' 00223 00226 00225 00226 0022(1 00221, 00227 18-12-76 21-01-75 10-03-75 15-06-75 21-05-75 01-11-73 11-07-76 21-01-75 10-03-75 15-06-75 21-05-75 01-11-73 01-11-73 11-07-76 18-12-76 22-01-75 21-05-75 01-11-73 30-07-76 19-12-76 21-11-73 11-07-76 19-12-76 21-01-75 12-03-75 20-05-75 01-11-73 01-11-73 29-07-76 18-12-76 21-01-75 10-03-75 20-05-75 01-11-75 29-07-76 18-12-76 21-01-75 10-03-75 20-05-75 01-11-73 01-11-73 01-11-73 01-11-/3 01-11-73 01-11-73 01-11-73 01-11-73 11-07-/6 18-12-76 01-11-73 0.065 0.052 0.053 0.066 O.O63 0.050 0.060 0.052 0.050 O.O69 0.068 0.022 0.068 0.081 0.062 0.066 0.067 0.066 0.050 0.051 0.050 0.066 0.052 0.067 0.052 0.065 0.023 0.062 0.067 0.036 0.029 0.035 O.O63 0.015 0.015 0.017 0.021 0.018 0.015 0.038 0.0)2 0.017 0.067 0.026 0.083 0.039 0.035 0.036 0.027 0.055 0.020 0.018 0.017 0.020 0.021 0.015 0.017 0.016 0.016 0.016 0.018 0.006 0.032 0.028 0.022 0.026 0.019 0.023 0.016 0.018 0.012 0.012 0.011 0.012 0.012 0.010 0.008 0.018 0.026 0.016 0.018 0.017 0.021 0.008 0.007 0.009 0.008 0.008 0.008 0.008 0.006 0.006 0.016 0.010 0.030 0.01 s 0.010 0.009 0.008 0.026 0.085 0.070 0.070 0.086 0.086 0.065 0.077 0.066 0.066 0.065 0.066 0.026 0.100 0.109 0.086 0.090 0.066 0.089 0.066 0.069 C.062 0.078 0.063 0.059 0.066 0.055 0.031 (.060 0.071 0.068 0.067 0.052 0.066 O.023 0.022 0.026 0.029 0.026 0.025 0.066 0.016 0.023 0.063 0,036 0.110 0.056 0.065 0.065 0.035 0.079 G1 HONS 056516 parts per million Col 1cction PCB PCB PCB w __ Date 1255 1260 Total 00228 00228 00228 00228 00228 00229 00229 00229 00229 00229 00229 00230 00231 00231 00231 00231 00231 00231 00232 00232 00232 00232 00232 00232 00250 00251 00251 00252 00252 00253 00255 00255 00255 00255 00256 00256 0025/ 00257 00257 00257 00257 00257 00257 00258 00258 00258 00258 00258 00258 00258 11-07-75 18-12-75 22-01-75 12-03-75 21-05-75 11-07-75 18-12-75 22-01-75 12-03-75 15-05-75 21-05-75 11-07-75 11-07-75 17-12-75 21-01-75 12-03-75 16-05-75 20-05-75 11-07-75 17-12-75 21-01-75 12-03-75 16-05-75 20-05-75 02-11-73 02-11-73 15-07-75 02-11-73 15-07-75 02-11-73 02-11-73 15-07-75 02-11-73 31-07-75 02-11-73 15-07-75 02-11-73 15-07-75 02-10-75 22-10-75 13-12-75 19-02-75 26-03-75 02-11-73 15-07-75 22-10-75 05-11-75 19-02-75 26-03-75 05-06-75 0.025 0.027 0.019 0.015 0.016 0.016 0.015 0.016 0.012 0.016 0.017 0.050 0.023 0.020 0.019 0.053 0.033 0.018 0.039 0.052 0.062 0.022 0.052 0.039 0.029 0.009 0.007 0.009 0.023 0.020 0.023 0.025 0.026 0.031 0.018 0.015 0.087 0.075 0.107 0.103 0.105 0.102 0.107 0.100 0.130 0.116 0.130 0.105 0.138 0.108 0.006 0.006 0.003 0.003 0.005 0.005 0.005 0.005 0.005 0.005 0.006 0.015 0.008 0.006 0.006 0.010 0.013 0.007 0.011 0.011 0.017 0.008 0.017 0.013 0.010 0.002 0.003 0.005 0.012 0.005 0.005 0.005 0.007 0.010 0.010 0.011 0.018 0.021 0.026 0.025 0.022 0.021 0,026 0.028 0.039 0.037 0.036 0.027 0.035 0.050 0.031 0.033 0.022 0.017 0.021 0.021 0.019 0.021 0.017 0.021 0.023 0.055 0.031 0.026 0.025 0.053 0.056 0.025 0.050 0.053 0.079 0.030 0.069 0.052 0.039 0.011 0.010 0.015 0.035 0.025 0.027 0.029 0.033 0.051 0.028 0.026 0.105 0.096 0.133 0. 128 0.127 0.123 0.133 0.128 0.169 0.153 0. 166 0.131 0.173 0.158 . parts per million Col lection PCB PCB PCB ip/y Date 1255 1260 To to I 00259 00259 00259 00259 00259 00259 00260 00261 00261 00261 00261 00261 00261 00261 00261 00262 00262 00262 00262 00262 00262 00262 00263 00263 00263 00263 00263 00263 00265 00265 00265 00265 00265 00265 00265 00265 00265 00265 00265 00265 00265 00265 00266 00266 00266 00266 00266 00266 00266 00267 02-11-73 15-07-75 15-01-75 19-02-75 26-03-75 05-06-75 02-11-73 02-11-73 15-07-75 02-10-75 13-12-75 15-01-75 19-02-75 26-03-75 05-06-75 02-11-73 15-07-75 13-12-75 15-01-75 19-02-75 26-03-75 05-06-75 02-11-73 15-07-75 15-01-75 19-02-75 26-03-75 05-06-75 02-11-73 15-07-75 11-12-75 15-01-75 19-02-75 26-03-75 05-06-75 02-11-73 15-07-75 13-12-75 15-01-75 19-02-75 26-03-75 05-06-75 02-11-73 31-07-75 13-12-75 15-01-75 19-02-75 26-03-75 05-06-75 02-11-73 0.179 0.193 0.153 0.255 0.165 0.170 0.067 0.036 0.081 0.053 0.053 0.055 0.052 0.055 0.066 6.035 0.030 0.036 0.055 0.055 0.032 0.036 0.185 0.121 0.139 0.095 0.152 0.115 0.027 0.016 0.018 0.028 0.016 0.015 0.022 0.119 0.091 0.128 0.112 0.096 0.150 0.096 0.092 0.081 0.076 o.m 0.105 0.096 0.076 0.065 0.070 0.066 0.076 0.087 0.080 0.096 0.027 0.016 0.036 0.019 0.019 0.020 0.017 0.018 0.023 0.012 o.on 0.012 0.012 0.016 0.010 0.011 0.062 0.066 0.065 0.038 0.066 0.067 0.010 0.008 0.007 0.008 0.008 0.007 0.013 0.032 0.032 0.037 0.033 0.029 0.060 0.031 0.032 0.029 0.021 0.033 0.036 0.036 0.028 0.025 0.269 0.259 0.227 0.331 0.266 0.266 0.096 0.050 0.117 0.062 0.072 0.076 0.059 0.063 0.089 0.067 0.061 0.068 0.057 0.060 0.062 O.O67 0.267 0.167 0.185 0.133 0 218 0 162 0 037 0 026 0.025 0.036 0 026 0.071 0.035 0.151 0.123 0.165 0.165 0.125 0.180 0.12? 0.126 0.110 0.097 0.166 0.1.33 0.178 0.102 0.090 62 HONS OS6515 parts per million Collection PCB PCB pee ID# Da te 1256 1260 Total 00367 00267 00267 00267 00267 00268 00268 00268 00268 00268 00268 00268 00268 0026? 00269 00269 00269 00269 00269 0026? 0026? 00269 00270 00271 00271 00272 00272 00272 00273 00273 00273 00276 00276 00276 00300 00301 00301 00301 00301 00302 00302 00302 00302 00302 00303 00306 00305 00305 00305 00305 31-07-76 11-12-76 15-01-75 26-03-75 06-06-75 02-11-73 15-07-76 02-10-76 11-12-76 15-01-75 19-02-75 26-03-75 06-06-75 02-11-73 15-07-76 22-10-76 05-11-76 11-12-76 15-01-75 19-02-75 26-03-75 06-06-75 02-11-73 15-07-76 06-06-75 31-07-76 19-02-75 06-06-75 15-07-76 11-12-76 06-06-75 15-07-76 11-12-76 06-06-75 13-11-73 13-11-73 26-06-76 10-01-75 12-02-75 13-11-73 10-01-75 12-02-75 09-06-75 28-05-75 10-01-75 13-11-73 10-01-75 12-02-75 09-06-75 28-05-75 0.062 0.058 0.063 0.069 0.068 0.050 0.068 0.053 0.056 0.055 0.062 0.066 0.057 0.023 0.020 0.017 0.018 0.020 0.018 0.018 0.016 0.016 0.026 0.015 0.012 0.072 0.077 0.060 0.016 0.021 0.019 0.162 0.120 0.130 0.051 0.036 0.023 0.028 0.023 0.071 O.O63 0.039 0.066 0.036 0.032 0.065 0.011 0.016 0.010 0.016 0.021 0.026 0.028 0.025 0.025 0.012 0.016 0.016 0.016 0.015 0.015 0.012 0.013 0.002 0.003 0,002 0.002 0.003 0.002 0.002 0.002 0,006 0.012 0.006 0.006 0.017 3.022 3.025 0.006 0.006 0.007 0.097 0.086 0.099 0.009 0,006 0.005 0.005 0.003 0.016 0.013 0.011 0.016 o.oto 0.011 0.012 0.002 0.003 0.003 0.011 0.083 0.082 0.091 0.096 0.093 0.062 0.066 0.069 0.070 0.070 0.077 0.056 0.070 0.025 0.023 0.019 0.020 0.023 0.020 0.020 0.018 0.020 0.038 0.019 0.016 0.089 0.099 0.085 0.022 0.027 0.026 0.239 0.206 0.229 0.060 0.038 0.028 0.033 0.026 O.O85 0.056 0.050 0.058 0.066 0.063 0.057 0.013 0.019 0.013 0.025 parts per nil!jon Col 1ectIon PCB pee PCB ID# Dote__ _ 1256 1260 Total 00306 00307 00309 00309 00309 00309 003)0 00310 00310 00310 00311 00311 00311 00312 00312 00312 00312 00312 00312 00313 00313 00316 00315 00315 00316 00316 00317 00317 00318 003 i 8 00319 00319 00320 00320 00321 00322 00322 00322 00323 00323 00323 00323 00326 00326 00326 00326 00325 00325 00325 00325 13-11-73 13-11-73 13-11-73 16-07-76 09-01-75 27-05-75 13-11-73 16-07-76 10-01-75 27-05-75 13-11-73 26-06-76 16-07-76 13-11-73 01-08-76 09-01-75 12-02-75 28-05-75 09-06-75 13-11-73 01-08-76 13-11-73 13-11-73 01-08-76 13-11-73 16-07-76 13-11-73 16-07-76 13-11-73 16-07-76 13-11-73 16-07-76 13-11-73 16-07-76 13-11-73 13-11-73 16-07-76 09-01-75 13-11-73 01-08-76 09-01-75 27-05-75 13-11-73 01-08-76 09-01-75 27-05-75 13-11-75 01-08-76 09-01-75 12-02-75 0,031 0.018 0.029 0.066 0.066 0.055 0.0)7 0.019 0.022 0.017 0.073 0.062 0.050 0.066 0.066 0.068 0.029 0.038 0.029 0.006 0.009 0.010 0.028 0.030 0.013 0.008 0.020 0.023 0.006 0.008 0.009 0.010 0.010 0.010 0.027 0.060 0.061 0.060 0.035 0.060 0.055 0.056 0.033 0.066 0.069 0.029 0.058 0.069 0.066 0.036 0.009 0.006 0.009 0.012 0.012 0.016 0.003 0.006 0.006 0.006 0.023 0.021 0.018 0.019 0.018 0.016 0.018 0.023 0.016 0.001 0.001 0.001 0.011 0.011 0.003 0.003 0.006 0.006 0.006 0.006 0.006 0.005 0.006 0.005 0.005 0.015 0.019 0.017 0.015 0.018 0.023 0.017 0.008 0.012 0.009 0.010 0.017 0.016 0.016 0.018 0.060 0.022 0.038 0.058 0.053 0.069 0.020 0.023 0.026 0.021 0.036 0.083 0.068 0.055 0.066 0.062 0.067 0.061 0.065 0.007 0.010 0.011 0.039 0.061 0.016 0.011 0.026 0.029 0.010 0.012 0.013 0.015 0.016 0.015 0.032 0.055 0.080 0.077 0.050 0.058 0.078 0.071 0.061 0.053 0.053 0.039 0.075 0.065 0.053 0.052 63 MOMS 056516 parts per ml 11 ion Col lection PC \i PCB PCB IDA ____Date___ 1254 1260 Total 00355 00355 00355 00355 00355 00356 00356 00356 00356 00356 00356 00357 00357 00357 00357 00358 00358 00358 00358 00358 00358 00359 00359 00360 00360 0036I 00361 00361 0036I 00361 00362 00362 00362 00362 00362 00362 00363 00363 00363 00363 00363 00366 00364 00364 00365 00365 00365 00365 00365 00365 16-07-74 09-01-75 12-02-75 09-04-75 28-05-75 16-07-74 09-01-75 12-02-75 24-04-74 09-04-75 28-05-75 24-04-74 16-07-74 09-01-75 20-05-75 24-04-74 16-07-74 09-01-75 12-02-75 09-04-75 28-05-75 24-04-74 01-08-74 24-04-74 01-08-74 24-04-74 24-04-74 17-07-74 10-01-75 12-02-75 24-04-74 16-07-74 09-01-75 12-02-75 09-04-75 28-05-75 24-04-74 01-03-74 12-02-75 09-04-75 27-05-75 24-04-74 01-08-74 28-05-75 24-04-74 16-07-74 10-01-75 12-02-75 09-04-75 28-05-75 0.032 0.040 0.033 0.033 0.135 0.047 0.048 0.053 0.051 0.043 0.049 0.015 0.018 0.013 0.019 0.100 0.091 0.1)9 0.108 0.097 0.106 0.026 0.025 0.017 0.020 0.031 0.028 0.045 0.028 0.029 0.025 0.025 0.024 0.027 0.019 0.025 0.026 0.040 0.025 0.021 0.025 0.006 0.008 0.012 0.011 0.011 0.008 0.007 0.010 0.009 0.011 0.012 0.011 0.010 0.038 0.017 0.019 0.020 0.020 0.016 0.019 0.007 0.008 0.008 0.009 0.042 0.040 0.058 0.044 0.038 0.041 0.007 0.008 0.002 0.003 0.012 0.011 0.020 0.012 0.013 0.009 0.009 0.007 0.009 0.008 0.007 0.010 0.021 0.014 0.011 0.015 0.001 0.003 0.002 0.003 0.004 0.004 0.005 0.003 0.004 0.043 0.052 0.044 0.043 0. 173 0.064 0.067 0.073 0.071 0.059 0.068 0.022 0.026 0.021 0.028 0. 142 0.131 0.177 0.152 0.135 0.147 0.033 0.033 0.019 0.023 0.043 0.039 0.065 0.040 0.042 0.034 0.034 0.031 0.036 0.027 0.032 0.036 0.061 0.039 0.032 0,040 0.007 0.011 0.014 0.014 0.015 0.012 0.012 0.013 0.013 parts per ni11 ion Col lection PCB PCB PCO IDA Date 1254 1260 Total 00366 00366 00366 00366 00366 00367 00367 00367 00368 00369 00369 00369 00370 00370 00370 00370 00370 00371 00371 00371 00371 00371 00372 00373 00373 00373 00373 00373 00374 00374 00374 10013 10029 10033 10035 10084 10085 10100 10110 10111 10118 10129 10130 10138 10150 10166 10168 10171 10172 II029 24-04-74 16-07-74 10-01-75 12-02-75 09-04-75 16-07-74 10-01-75 28-05-75 16-07-74 17-07-74 IO-OI-75 12-02-75 01-08-74 10-01-75 11-02-75 08-01-75 27-05-75 01-08-74 10-01-75 11-02-75 08-04-75 27-05-75 01-08-74 01-08-74 09-01-75 12-02-75 09-04-75 28-05-75 10-01-75 12-02-75 09-04-75 07-11-73 07-11-73 07-11-73 07-11-73 07-11-73 07-11-73 07-11-73 07-11-73 07-11-73 07-11-73 07-11-73 07-11-73 07-11-73 07-11-73 07-11-73 07-11-73 07-11-73 07-11-73 07-11-73 0.015 0.020 0.016 0.014 0.016 0.057 0.074 0.057 0.013 0.028 0.027 0.027 0.023 0.023 0.030 0.036 0.022 0.018 0.028 0.019 0.019 0.028 0.031 0.040 0.049 0.036 0.041 0.044 0.040 0.037 0.039 0.007 0.020 0.023 0.009 0.032 0.013 0.029 0.016 0.018 0.016 0.022 0.012 0.024 0.009 0.021 0.020 0.014 0.024 0.018 0.003 0.003 0.005 0.002 0.003 0.036 0.049 0.029 0.010 0.008 0.009 0.007 0.011 0.013 0.011 0.020 0.010 0.007 0.008 0.006 0.007 0.009 0.014 0.024 0.027 0.023 0.023 0.024 0.005 0.004 0.004 0.004 0.003 0.007 0.003 0.010 0.004 0.009 0.004 0.003 0.005 0.008 0.007 0.008 0.004 0.004 0.006 0.003 0.009 0.008 0.018 0.023 0.021 0.016 0.019 0.093 0.123 0.086 0.023 0.036 0.036 0.034 0.034 0.036 0.041 0.056 0.032 0.025 0.036 0.025 0.026 0.037 0.045 0.054 0.076 0.059 0.064 0.063 0.045 0.041 0.043 0.011 0.023 0.030 0.012 0.042 0.017 0.038 0.320 0.021 0.02) 0.030 0.019 0.032 0.313 0.0?5 0.026 0.017 0.033 0.026 64 HONS 05651? parts per million Collection PCB PCB PCB ID# Date ' 1256 1260 Tota 1 00325 00325 00326 00326 00326 00326 00326 00326 00327 00328 00328 00328 00328 00328 00328 00329 00330 00330 00330 00330 00530 00330 00330 00331 00331 00331 00331 00331 00331 00331 00332 00332 00332 00333 00333 00333 00336 00336 00336 00336 00335 00335 00335 00335 00335 00336 00336 00336 00336 00336 09-06-75 28-05-75 13-11-73 01-03-76 09-01-75 12-02-75 09-06-75 28-05-75 13-11-73 13-11-73 26-06-76 16-07-76 09-01-75 12-02-75 27-05-75 13-11-73 13-H-73 26-06-76 16-07-76 10-01-75 12-02-75 09-06-75 28-05-75 13-11-73 26-06-76 16-07-76 10-01-75 12-02-75 09-06-75 28-05-75 09-01-75 09-06-75 28-05-75 09-01-75 09-06-75 28-05-75 13-11-73 01-08-76 12-02-75 27-05-75 13-11-73 01-08-76 12-02-75 09-06-75 27-05-75 13-11-73 16-07-76 12-02-75 09-06-75 27-05-75 0.056 0.051 0.033 0.066 0.053 0.061 0.035 0.056 0.017 0.032 0.025 0.033 0.062 0.039 0.027 0.017 0.020 0.031 0.030 0,028. 0.036 0.025 0.031 0.103 0.083 0.098 0.075 0.098 0.085 0.121 0.035 0.027 0.066 0.029 0.025 0.026 0.029 0.039 0.036 0.038 0.035 0.033 0.066 0.053 0.063 0.025 0.026 0.061 0.019 0.026 0.018 0.016 0.011 0.015 0.016 0.016 0.016 0.018 0.005 0.015 0.012 0.015 0.016 0.017 0.015 0.005 0.010 0.013 0.012 0.012 0.011 0.011 0.016 0.066 0.032 0.036 0.026 0.062 0.036 0.051 0.016 0.013 0.017 0.009 0.007 0.007 0.011 0.016 0.015 0.016 0.008 0.008 0.012 0.013 0.016 0.012 0.013 0.017 0.010 0.010 0.076 0.065 0.066 0.059 0.069 0.057 O.O69 0.076 0.022 0.067 0.037 0.068 0.058 0.056 0.062 0.022 0.030 0.066 0.062 0.060 0.065 0.036 0.065 0.152 0.115 0.136 0.101 0.160 0.121 0.172 0.051 0.060 0.061 0.038 0.032 0.031 0.060 0.053 0.051 0.052 0.063 0.061 0.056 0.066 0.077 0.037 0.037 0.058 0.029 0.036 . parts per million Col lection PCB PCB PCB ID# Date JL116 1260 TotaJ_ 00337 00337 00337 00337 00337 00338 00338 00038 00338 00338 00338 00339 00339 00339 00339 00339 00360 00361 00362 00362 00363 00363 00366 00365 00366 00366 00367 00367 OO360 00369 00350 00350 00351 00351 00351 00352 00352 00352 00352 00352 00352 00353 00353 00353 00353 00353 00353 00356 00356 00355 13-11-73 16-07-76 12-02-75 09-06-75 27-05-75 13-11-73 01-08-76 09-01-75 12-02-75 09-06-75 27-05-75 13-11-73 16-07-76 09-01-75 12-02-75 28-05-75 13-11-73 13-11-73 13-11-73 01-08-76 13-11-73 16-07-76 13-11-73 13-11-73 13-11-73 16-07-76 13-11-73 16-07-76 13-11-73 13-11-73 09-01-75 28-05-75 13-11-73 09-01-75 28-05-75 13-11-73 16-07-76 10-01-75 19-02-75 16-06-75 27-05-75 13-11-73 16-07-76 10-01-75 19-02-75 16-06-75 27-05-75 13-11-73 01-08-76 26-06-76 O.023 0.03! 0.033 0.032 0.032 0.081 0.079 0.052 0.052 0.061 0.060 0.058 0.056 0.061 0.063 0.061 0.036 0.026 0.011 0.013 0.010 0.016 0.009 0.015 0.013 0.016 0.012 0.018 0.033 0.016 0.076 0.088 0.059 0.061 0.068 0.070 0.076 0.068 0.072 0.062 0.095 0.038 0.068 0.058 0.053 O.O63 0.052 0.030 0.023 0.033 0.005 0.028 0.010 0.061 0.007 0.060 0.010 0.062 0.007 0.039 0.033 0.116 0.061 0.120 0.026 0.078 0.029 0.081 0.036 0.095 0.028 0.088 0.025 0.083 0.018 0.072 0.025 0.086 0.025 0.088 0.021 0.082 0.007 0.061 0.003 0.027 0.007 0.018 0.009 0.022 0.006 0.016 0.005 0.019 0.006 0.015 0.006 0.019 0.006 0.017 0.005 3.019 0.003 9.015 0.006 0.022 0.012 0.065 0.006 0.020 0.032 0.105 0.033 0.121 0.022 0.0 31 0.019 0.020 0.026 0.002 0.023 O.C03 0.029 0.103 0.022 0.0-;n 0.025 0.0)7 0.026 O.i ; 0.035 0. r0 0.010 ,0.0-3 0.015 0.- ,J 0.016 0.0/' 0.01/ 1 0 0.011 0.0 Vt 0.016 0.6 .0 0.003 C '.. ' 1 0.006 0.0.7 0.003 o.c.i 05 HONS 056518 APPENDIX B LABORATORY DATA FOR PCB TESTS UTILIZING PRELIMINARY ANALYTICAL PROCEDURES 66 MONS 056519 TABLE 19. BLOOD SAMPLES TESTED BY THE DEGRADATIONDERIVATIZATION TECHNIQUE1 Traverse City Sample ID# 00300* 00302* 00303* 00305* 00332'"' 00333** 00350* Total PCB as Decachloroblphenyl in Parts Per Ml 11 Ion Nov. 1973 Apr. 1976 Sample Date May 1976 6-25-76 6-26-76 July 1976 0.015 0.025 0.026 0.015 0.028 0.020 0.016 0.006 0.032 0.008 0.023 0.017 0.017 0.016 0.0)5 0.005 0.018 0.025 0.006 0.017 Manistee Sample IDIf 00257 00258 00261 00262 00268 00269 Total PCB as Decarhloroblphcnyl In Parts Per Ml 11 Ion 10-2-76 Sample Date 10-2-76 Nov. 1976 0.075 0.060 0.032 0.022 0.038 0.008 0.076 0.072 0.030 0.036 0.036 O.O67 0.079 O.O67 0.016 + Recovery of decachlorobiphenyl by this technique was &$% * Single extraction used ** Triple extraction used Accuracy questionable 67 MONS 056520 APPENDIX C PHYSICAL DATA FOR STUDY PARTICIPANTS 08 HONS 056521 TABLE 20. PARTICIPANT PHYSICAL DATA - LUDINGTON I.D.# 00200 00201 00202 00203 00201 00205 00206 00207 00208 00209 00211 00210 00212 00213 00219 00215 00216 00217 00218 00219 00220 00221 00222 00223 00229 00225 00226 00227 00228 00229 00230 00231 00232 00233 Sex M r F 'F F M F F F F F M F M M F F M F M F M M M H M F M M F F F M M Age . 51 99 38 28 58 27 23 69 60 53 98 50 90 38 99 .93 91 93 93 60 99 18 26 53 39 36 38 37 33 29 31 33 92 Height Weight 71 61 63 66 62 68 69 66 68 63 63 66 69 72 72 62 69 79 61 72 69 73 71 71 70 ' 65 70 67 62 69 72 72 f 200 155 159 135 220 158 125 165 192 197 192 150 130 178 160 120 197 208 115 199 150 170 17 210 170 155 160 225 125 102 205 213 HONS 056522 69 TABLE 20. PARTICIPANT PHYSICAL DATA MAM I STEP I.D.* 00250 00251 00252 00253 00254 00255 00256 00257 00258 00259 00260 00261 00262 00263 00264 00265 00266 00267 00268 00269 00270 00271 00272 00273 00274 Sex M F M F F F M F M M F M F M F M M r M F M F M F M Height WgiGllt 68 62 38 70 43 67 43 6 5 59 62 54 63 62 62 53 65 65 72 66 73 64 65 56 69 54 07 51 76 51 61 23 75 62 67 59 63 43 69 36 65 45 69 42 64 45 69 49 64 48 68 180 152 143 138 120 148 115 140 180 208 14 4 200 140 234 114 230 167 123 165 230 160 153 185 180 + 155 70 MONS 056523 TABLE 20. PARTICIPANT PHYSICAL DATA - TRAVERSE CITY I.D.# 00300 00301 00302 00303 00304 00305 00306 00307 00308 00309 00310 00311 00312 00313 00314 00315 00316 00317 00318 00319 00320 00321 00322 00323 00324 00325 00326 00327 00328 00329 00330 00331 00332 00333 00334 00335 00336 00337 00338 00339 00340 00341 00342 00343 00344 00345 00346 00347 00348 00349 00350 Sex F F F F F F M F F M F M r F F M F F M M F F F M F F M F F M F M M F M F M F M F M M M F M F M F M F M Age 78 50 63 67 73 41 56 53 64 38 36 53 55 21 .58 33 56 26 43 22 56 47 31 31 58 58 52 41 57 47 50 58 52 41 41 63 56 57 69 60 60 27 27 23 23 53 51 29 26 42 Height Weight 58 140 66 155 70 1C5 64 14 2 60 125 69 172 68 175 63 222 75 220 69 200 70 225 60 130 57 71 176 67 130 67 145 71 147 72 165 65 130 62 120 64 145 69 165 65 180 61 140 68 152 61 62 140 66 160 62 120 67 180 69 175 67 135 69 125 05 135 70 170 05 212 72 179 65 139 70 170 66 135 71 160 68 138 72 170 64 125 65 140 62 110 71 160 66 117 70 185 HONS 056524 TABLE 20. (continued) U2JL 003S1 00352 00353 00354 00355 00356 00357 00358 00359 00360 00361 00362 00363 00364 00365 00366 00367 00368 00369 00370 00371 00372 00373 00374 Sex F M F F F M F M M F M F M F M F M M F M F F M F Age . 40 45 44 51 48 47 54 69 36 34 ' 62 66 28 22 64 62 58 43 58 35 35 38 58 21 Height 65 69 67 67 66 74 66 67 74 62 68 67 69 65 68 67 67 70 63 70 64 61 70 66 Weight 135 162 140 130 165 220 ieo 160 300 160 155 155 170 170 165 158 180 170 125 180 120 118 150 130 72 HONS 056525 TABLE 20. PARTICIPANT PHYSICAL DATA - SOUTH HAVEN E-p-y 00035 0003G 00053 00055 00062 00064 00065 00066 00067 00070 00071 00073 00106 00105 00114 00115 00120 00122 00124 00126 00128 00130 00116 00104 00140 00139 00119 00111 00121 00116 00072 _ss*_ M F M M M M M F M M F M F M M F F M M r M F M M F M M M M M F 28 24 23 51 53 44 34 39 57 29 48 36 37 45 47 '50 46 55 39 62 59 41 35 61 61 45 33 38 41 48 Heip.ht 72 165 65 125 72 195 71 164 73 185 70 174 72 165 65 115 72 180 72 162 63 120 67 169 66 160 77 210 72 63 110 68 135 70 222 72 126 71 192 65 160 71 175 72 172 66 140 68 150 70 185 70 200 70 165 70 175 67 140 73 HONS 056526 TABLE 20. PARTICIPANT PHYSICAL DATA - ALGONAC I.D. 10013 10029 11029 10033 10035 10100 10110 10111 10118 10129 10130 10138 10150 10166 10168 10169 10170 10084 10085 Sex F F M M M M M F M M F M M F F M M M M Age 51 59 55 72 66 52 34 31 68 64 50 73 33 48 48 56 71 69 35 Height 68 68 68 70 67 74 71 66 69 65 62 68 72 67 66 66 70 68 67 Height 128 185 161 176 161 220 220 125 220 155 107 193 188 146 242 170 160 150 180 HONS 05652? APPENDIX D SELECTED ILLUSTRATIONS OF THE CONCENTRATION OF PCB IN THE BLOOD OF PARTICIPANTS UHO CONSUMED GREAT LAKES FISH DURING THE STUDY . HONS 056528 7S APPENDIX D Illustrations of the correlation between PCB levels in blood and time for persons consuming 26 or more pounds of Great Lakes fish per year. PCB values from Tables 4, 5 6 6. Legend for Figures 10 and 11. Participant 00323: Male 31 years, 1974 weight 165 lbs., 1973-75 mean annual fish consumption 64 lbs./yr. Participant 00328: Male 41 years, 1974 weight 140 lbs., 1973-75 mean annual fish consumption 58 lbs./yr. Participant 00336: Male 63 years, 1974 weight 170 lbs., 1973-75 mean annual fish consumption 35 lbs./yr. Participant 00213: Male 38 years, 1974 weight 178 lbs., 1973-75 mean annual fish consumption 132 lbs./yr. Participant 00324: Female 31 years, 1974 weight 180 lbs., 1973-75 mean annual fish consumption 29 lbs./yr. Participant 00226: Female 36 years, 1974 weight 155 lbs., 1973-75 mean annual fish consumption 132 lbs./yr. Participant 00312: Female 55 years, 1974 weight 130 lbs., 1973-75 mean annual fish consumption 49 lbs./yr. Participant 00363: Male 28 years, 1974 weight 170 lbs., 1973-75 mean annual fish consumption 65 lbs./yr. Participant 00557 Female 54 years, 1974 weight 180 lbs., 1973-75 mean annual fish consumption 190 lbs./yr. Participant 00309 Male 38 years, 1971. weight 220 lbs., 1973-75 mean annual fish consumption 42 lbs./yr. Participant 00337: Female 56 years, 1974 weight 212 lbs., 1973-75 mean annual fish consumption 33 lbs./yr. Participant 00310: Female 36 years, 1974 weight 200 lbs., 1973-75 mean annual fish consumption 22 lbs./yr. ic HONS 056529 U. 1VU 0.100k _I_________ January 1974 X-. July ___ - ..1 January 1978 _____________ L July PC8 LEVEL IN BLOOD (ppm ) January 197M July TIME January 1978 July FIGURE 10 . CHANGE IN BLOOD PCB LEVELS OVER TIME FOR PERSONS , EXPOSED TO LAKE MICHIGAN FISH 77 HONS 056530 0.120 0.100 0.080 0.060 00312 0.040 0.020 00363 00357 III.............................................. ... ..........L January 1974 July ' January 1975 July ' 0.120 - 0.100 - PCB LEVEL IN BLOOD (ppm ) 0.080 0.060 0.040 0.020 1I ................................. ..........1IL January 1974 July January 1975 TIME July FIGURE 11. CHANGE IN BLOOD PCB LEVELS OVER TIME FOR PERSONS EXPOSED TO LAKE MICHIGAN FISH 70 HONS 056531 APPENDIX E SELECTED ILLUSTRATIONS OP THE CONCENTRATION OF PCB IN THE BLOOD OF PARTICIPANTS WHO ABSTAINED FROM EATING GREAT LAKES FISH DURING 1975 7 9 MONS 056532 APPENDIX E Illustrations of the correlation between PCB levels in blood and time for persons who abstained from consuming Great Lakes fish for various periods of time. PCB values from Tables *4 , 5 6 6. Legend for Figures 12 and 13. Participant 00338: Male 57 years, 1974 weight 179 lbs., 1973-75 mean annual fish consumption 80 lbs./yr. Participant 00312: Female 55 years, 1974 weight 130 lbs., 1973-75 mean annual fish consumption 49 lbs./yr. Participant 00210: Male 50 years, 1974 weight 150 lbs., 1973-75 mean annual fish consumption 24 lbs./yr. Participant 00330: Female 47 years, 1974 weight 120 lbs., 1973-75 mean annual fish consumption 62 lbs./yr. Participant 00352: Male 45 years, 1974 weight 162 lbs., 1973-75 mean annual fish consumption 30 lbs./yr. Participant 00356: Male 47 years, 1974 weight 220 lbs., 1973-75 mean annual fish consumption 100 lbj./yr Participant 00355: Female 48 years, 1974 weight 165 lbs., 1973-75 mean annual fish consumption 28 lbs./yr. Participant 00267: Female 59 years, 1974 weight 123 lbs., 1973-75 mean annual fish consumption 24 lbs./yr. Participant 00217: Male 43 years, 1974 weight 208 lbs., 1973-75 mean annual fish consumption 24 lbs./yr. Participant 00218: Female 43 years, 1974 weight 115 lbs., 1973-75 mean annual fish consumption 10 lbs./yr. 80 MONS 056533 PCB LEVEL IS BLOOD <ppm> 0.120 0.100 0.080 0.060 00352 00356 0.040 0.020 00355 Abstinence Periods 0.120 ---- J---------------------------- 1----------------------------- 1L January July ' January 1974 1975 July 0.100 0.080 0.060 0.040 0.020 January 1974 July TIME January 1975 July FIGURE 12. CHANGE IN BLOOD PCB LEVELS OVER TIME FOR PERSONS WHO ABSTAINED FROM LAKE MICHIGAN FISH CONSUMPTION HONS 056534 V.LnJ 0.100 0.080 0.060 0.040 0.020 .. 1.............................._________ January 1974 July .. . ___1 January 1975 ______ 1 July PCB LEVEL IN BLOOD (ppm) TIME FIGURE 13. CHANGE IN BLOOD PCB LEVELS OVER TIME FOR PERSONS WHO ABSTAINED FROM LAKE MICHIGAN FISH CONSUMPTION MONS 056535 BIBLIOGRAPHY 1. Risebrough, R.W.i et al, "Polychlorinated Biphenyls in Global Ecosystem," Nature 220, 1098, (1968). 2. Env. Hlth. Persp. 1, 3-185, (1972). 3. Price, H.A., 6 R.L. Welch, "Occurrence of Polychlorinated Biphenyls in Humans." F.nv. Hlth. Persp. 1 , 7 3-7 8 , (197 2 ) 9. Yobs, A.R., "Levels of Polychlorinated Biphenyls in Adipose Tissue of the General Population of the Nation." Env. Hlth. Persp. 1, 79-80, (1972). 5. Kimbrough, R.D., "The Toxicity of Polychlorinated Polycyclic Compounds and Related Chemicals." CRC Crlt. Rev. Toxicol. 2, 445-498, (1974). 6. National Conference on PCB's, November 19-21, 1975, Chicago, Illinois. 7. Kuratsune, M., T. Yoshimura, <J. Matsuyaka, 6 A. Yamaguchi, "Epidemiologic Study on Yusho, A Poisoning Caused by Ingestion of Rice Oil Contaminated with a Commercial Brand of Polychlorinated Biphenyls." Env. Hlth. Persp. 1, 119-128, (1972). ' 8. Federal Register 38 (129), 18096-18103, (1973). 9. Federal Register 41 (39), 8409-8410, (1976). 10. Allen, J.J!., D.H. Norback, 6 I.C. Hsu, "Tissue Hodificaticns in Monkeys as Related to Absorption, Distribution, and Excretion of Polychlorinated Biphenyls." Arch,, Env ConJ^om^ 2 , 86-92 , (1974). 83 MOMS 056536 11. Kimbrough, R.D. , R.E. Linder, V.W. Burse, 6 R.W. Jennings, "Adenofibrosis in the Rat Liver." Arch Env. Hlth. 27 390-395, (1973). 12. Kuratsune, M. , Y. Masuda, J. Naguyama, "Some Recent Tindings Concerning Yusho." Presented at the National Conference on PCB's, November 19-21, 1975, Chicago, Illinois. 13. Barsotti, D.A., $ J.R. 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E. Zabik, "Effects of Cooking on Concentrations of PCB and DDT Compounds in Chinook and Coho Salmon from Lake Michigan." J, Fisheries Res. Brd of Canada (5), 702-706 , 1973. 20. Hutzinger, 0., S. Safe, and V. Zitko, Th Chemistry of PCB1s, CRC Press, Ino., Cleveland, Ohio, 1974. 21. Thompson, J. F., ed., Analysis of Pesticide Residues in Human and Environmental Samples, See. 5, A, (3)(a), E.P.A., Research Triangle Park, N.C., (1972). 22. Burke, J. A., Division of Chemistry and Physios, F.D.A., Washington, D.C., private communication, August 1973. 23. Henderson, S. Of., J. G. DeBoer, and H. M. Stahr, Anal Chem. 43, 445--***47, (1971); and Griffith, F. D. , and R. V. Blanks, J.A.O.A.C. 57, 595-603, (1974). 2M. Henry, Richard J,, M.D., Clinical Chemistry: Principles and Techniques, Hoeber Medical Div., Harper E Row, 832, (1964). 25. Armour, J. A., J. A. Burke, J.A.O.A.C. 53, 761-768, (1970). 26. Berg, 0. W., P. L. Diosady, and G. A. V. Rees, Bull Environ. Contam. Toxicol. 7, 338-347, (1972). 27. Burke, J. A., Division of Chemist-y and Physics, F.D.A., Washington, D.C., private communications, February 1974 . 28. Young, S. J. V., and J. A. Burke, Bull Environ Contam. Toxicol. 7, 160-167, (1972). 85 MONS 056638 29. Mulhern, B. M. , et al., J.A.0.A.C. 54 , 548-550 , (1971). 30. Trotter, W. J., Division of Chemistry and Physics, F.D.A. Washington, D.C., private communication, August 1874 31. Goerlitz, D. r., and L. M. Law, JA.OA.C 57, 175-181, (1974). 32. Armour, J. A., J.A.O.A.C. 56, 987-993, (1973). 33. Huckins, J. N. , J. E. Swanson, and D. L. Stalling, J.A.O.A.C. 57, 416-417, (1974). 86 MONS 056539 PBB-PCB BIBLIOGRAPHY Aftoxmls, J.G., et al. The Toxicoloqy of Brominated Biphenyls 1. 0ra 1 Toxicity and Embryotoxicity, 11T Skin, Eye and Inhalation Toxicity and an Acut Test MethodJIT Body Tat. Presented at tTuTSociety of Toxicology HeitTngs at Williamsburg, Virginia, (March 8, 1972). Allen. J. R. Response of the Honhuman Primate to Polychlorinated Biphenyl Exposure. Tederation Proceedings, 34: TOT-IW, (dWT 1975). Allen, O.R. and Horback, U.H. Atsuko, Yamaguchi "Pathobiological Responses of Primates to Polychlorinated Biphenyl Exposure." To be published In Proceedings of the national Conference on Polychlorinated Biphenyls, (November 19, 1975). Investigaten Concerning Babies Born From Women Who Consun)e3~0|TCont*aiiiinated'Tilth Clilorobi phenyl . FukuokaTgakkai "Shi Fukuoka Acta MecTi ca, 62:1, 117 122, (1971). Translation by Leo Warner Associates. (Included in MDPH's PBB in Human Breast 1-ii 1 k: Background Material) ' Bernstein, I.A., Hopwood, M.L., "PBB Scientific Advisory Panel Report to William lrey, N.S,, Oehme, F W., Tephly, G. Hi 1Uken, Governor of Michigan on Polybrominated T., VanDuuren, B.L. Biphenyls (PBB)", (May 24, 1976). Biros, Francis J., et al. Polychlorinated Biphenyls in Human Adipose Tissue FuYTetTrT of EnvIronmentarTontamlnatron and Toxfcolo Vol. 5, No. 4: 317-323, (1970). Carter, Luther 0. Michigan's PBB Incident: Chemical Hix-up leads to Disaster.- Science 192:240-243, (April lb, i'976). Corbett, T.H., et al. Toxi cl ty of PolybVominated Biphenyls (fi romcster BP-fFIn Rodents. EnvironmentaTResearch 10:35039B7(t575). ~ MOWS 056540 Courtney, K. Diane and Moore John A. Curley, A., et al. ` Curley, A., et al. . Deatrlck, Richard W. Dent, O.G., Netter, K.O., and Gibson, J.E. Ficsor, Gyala and Wertz, Gary F. Fllonow, A.B., Food and Drug Administration, Bureau of Veterinary Medicine. -1- Teratoloqy Studies with 2,4.5Trichlorophenox.yacetic Acid and 2.3.7,8Yetrachlorodibenzo-P. dioxin. foxicoloov an<T Appl led"Pharmacology 20; 396-403 (1971) Polychlorinated Biphenyls: Distribution Storaqe in "Body Fluids and Tissue's of. ' Sherman Rats. Environmental Research. Volume 4, number 6, (December, 1971). Polychlorinated Biphenyls: Evidence of Transplacental Passaqe is the Sherman Rat. TtfTCosmet. loxTcoirTolTTl: '47T-47S7' (1973). "Long Term Human Health Implications of PBB Exposure." Memo to James Terrlan, M.D. (January 21, 1976). Effects of Chronic Administration of Polybromlnated Biphenyls on Parameters Associated with Hepatic Drug Metabolisn. Res.. Common, them. Pathol. Pharmacol, F3: 75-82, (1976). "Polybrom'nated Biphenyl Honteratogenic, C-Mitosis Synergist in Rat. Abstract" Presented at Environmental Mutagen Society Meeting, (March, 1976). To be published In Mutation Research. Jacobs, L. W. , and Mortland, M.M. "Fate of Polybrominated Biphenyls (PBBs) in Soils: II. Retention of llexa bromobiphenyl in Four Michgan Soils." For Submission to Journal of Agriculture Food Chem., (April, 1976). "Michigan State Dairy Herd Survey - A Report on Herd Health Status of Animals Exposed to Polybromlnated Biphenyls (PJ3)." Division of Veterinary Research, Beltsvillo, Maryland (April 25, 1975). HONS 056541 -3- fries,`George F. Fries, G.F,, et. al. Garthoff, L.H., et. al. Getty, S.H. and Hillman, D. Goldstein, J.A,, Hickman, P. and due, D.L. Hillman, D., Bolenbaugh, D., and Convey, E.M. Hooting, Alan L. "Estimates of Possible Human Exposure to PBB in Michigan." Pesticide Degradation Laboratory, Agricultural Environmental Quality Dlstitute, Beltsvi1le Agricultural Research Center, Beltsvilie, Maryland. IIfeat_M_!iixrQSQEilJj!tym2_Iiid.u.cin3 ruai_on J3Pl_.axidJieldjJja_iimijialLQn from Cows. Journal of Dairy Science, 54(3):364-368, (1971). "Biochemical Changes Caused by Ingestion of Arochlor 1254 (A Commercial Polychlorinat Biphenyl Mixture) or Firemaster BP-6 (A Commercial Polybrominated Biphenyl Mixture). U.S. food and Drug Administration Official Report. "An Investigation of a Suspected . Polybrominated Biphenyl (PBB) Toxicosis in the Sdington Herd of Dairy Cottle: Final Report." Departments of Large Aninnl Surgery and Medicine and Dairy Scieice; Michigan State University, (April 21, 1976). Experimental Hepatic Porphyria Induced by Polychlorinated biphenyls. Toxicol >qy and /Applied Pharmacology 27 : 437-448, (1974) ' "Iodine Toxicity and Thyroid Disturbance in Dairy Cattle." Department of Dairy Science, Michigan State University, East Lansing, Michigan, (1976). "The Michigan PBB Problem." Presented at the Spring Meeting of the Central States Association of Food and Drug Officials, St, Joseph, Michigan, (May 8, 1975). HONS 056542 Jackson, T.F. and Halbert, F.L. Jacobs, L.W., Chou, S.F. and Tledje, J.M. Kimbrough, Renate D, Kimbrough, Renate D. Kimbrough, R.O., et al. Kimbrough, R.D., et al. Kimbrough, R.D., et al. -4 - A Toxic Syndrome Associated with the Feeding of Pol'ybrominated BiphenylContaminated Protein Concentrate to Dairy Cattle. Journal of the American Veterinary Medical Association, Vol. 165, No. 5: 437-439, (1974). "Fate of Polybromlnoted Biphenyls (PBBs) in Soils: I. Persistence and Plant Uptake." For Submission to Journal of Agricultural Food Chem. , (April, 1976). "Pathological Findings Associated with Chronic Experimental Exposure to PCBs." Toxicology Branch; Center for Disease . Control, U.S. Public Health Service; Department of Health, Education, and Welfare; Atlanta, Georgia; (November 7, 1975). The Toxicity of Polychlorinated Polycyclic Compounds and Related Chemicals. Reprinted from the Toxicity of Polychlorinated Polycyclic Compounds and Related Chemicals. Critical Review Toxicology, Volume 2, Issue 4: 445-198, (1974). Induction of Liver Tumors In Rats by Polychlorinated Biphenyl Arochlor 1260. Journal of the National Cancer institute, 55: ' 1455-1459, (1976). Liaht Microscopy and Ultrastructure of Liver of Rats Fed Polych 1_orJnated_Bijhen_yis. Abstract'. Toxicology'and Applied Pharmacology 22(2): 315-316, (June, 1972). Morphological Changes in Livers of Rats fed T5Ty^^ na teOi phdnyns^LlgluTirb ro fc, Vy and ultnistructure. Archives o't IhvT:., Inta' "andTUTfrastructure. Health, Volume 25: 354 364, (November, 1972). HONS 056543 Lee, K.P., et al. Lee, K.P., et al. Meester, Walter D. Michigan Chemical Corporation. Michigan Department of . Agriculture Michigan Department of Agriculture Michigan Department of Agriculture Michigan Department of Natural Resources Michigan Department of Public Health -5 - Bromine Tissue Residue and Hepatotoxfc Effects of Octa bromo biphenyl in Rats. Toxicology and Applied Pharmacology, Volume 34: US127, (1975). Octa bromo biphenyl-induced Ultrastructural Changes In Rat liver. Archives of EnvironrnenTafliedlth", 31 : 465-471, (September 1975). "Critique on the Michigan Department of Public Health Stuc(y on the Short-term Effects of PBB on Health." (June 6, 1975. "Report on Operator Exposure to firemaster . B-p6 During Production." (June 12, 1975). "State of Michigan-Environmental Impact Statement - Disposal of Polybromlnated Biphenyl (PBB)-Contaminated Cattle in Kalkaska County," (August, 1974). Hearing lranscript (May 29, 1975). ' Hearing Transcript (June 10, 1976). "Technical Environmental Review DocumentPBB Contaminated Animal Disposal Site in Oscoda County, Michigan." Resource Recovery Division, (August 17, 1976). "Alleged Presence of Additional Toxic Factors in Farm Bureau Services Feed." HONS 0565** Michigan Department of Public Health. Michigan Department of Public Health ' Michigan Department of Public Health. Michigan Department of Public Health Michigan Department of Public Health Michigan Department of Public Health Moore, R.W., Dannan, C. and Aust, S.D. 6- ' "An Epidemiological Study of Human Health Effects Associated with the Consumption of Milk, Meat, or Other Foods Contaminated with Polybrominated Biphenyl-Addendum to Original Response to R.F.P. #223-75-2250. (Later became part of Center for Disease Control Contract #200-76-0625). "Evaluation of Changes of the Level of Polychlorinated Biphenyls (PCB) In Human Tissue." Final Report on FDA Contract #223-73-2209, (1976); "Knowledge Gained in the Six Months Since ' the Michigan Department of Public Health's Short Term Effects Report." P8B in Human Breast Milk: Background Material, (August 18, 1976). "Polybrominated Biphenyls: An Agricultural Incident and Its Consequences-An Epidmniolcgic Investigation of Human Exposure." Presented at 9th Annual Conference on Trace Substances in Environmental Health, (June 9-12, 1975). "Summary Comment on Michigan Department of . Public Health Study of Individuals Exoosed to PBB." Induction of Drug Metabolizing Enzyme; 1 n Rats Pursing from Mothers Fed PoVybronin'a'ted TSTphehyTs. Abstract .'"""Feci Proceedings, 35: 708(1976). HONS 056545 Norris, O.M., et al. O'Keefe, Patrick PBB Research Council Preache, M. and Gibson, J.E. Ringer, Robert K. Scrlpps, Stan Senate Special Investigating Committee , Vos, O.G., et al. -7- Toxicology of Octabromobiphepyl and Decabromodipheny1 Oxide. Environmental Health Perspectives, 11 : 153-161 , (1975) ' ''Preliminary Report-Analysis of Environmental Samples for Trace Components from Polybrominated Biphenyl Fire Retardants" (January, 1976). Letters of Response to Governor Hilliken's Request for Information Relative to the ' PBB Problem. "Protocols for Teratology, Prenatal ExposureBehavlorial Testing, Postnated Exposure ' Behavlorial Testing and Pre-and Postnatal Exposure-Behavioral Testing." Conclusions and Data "The Biological Performance of Chickens ' Following Polybrominated Biphenyl Administration in the Feed." Michigan State University, (1976). "Polybrominated Biphenyl-An Overall View." Michigan State University, (1975). "The Contamination Crisis In Michigan: Polybrominated Biphenyls." A Report from the Senate Special Investigating Committee on Polybrominated Biphenyls, (July, 11)75). Identification and Toxicological Evaluation of Chlorinated Dllienzofuran and Chlorinated Naphthalene in Two Commercial Polychlorinated Biphenyls Fd. Cosmet. Toxicol., 8:6:;5-623 .TlSToTT" MQNS 056546 Willett, t ,B. Willett, L B. and Irving H.A. . .Willett, L B. and Irving, H.A. Yobs, Anne R. Yoshimura, Takesumi . -8- "Polybrominated Biphenyls: A Study of Metabolism and Clearance In the Bovine." Specific Protocol and Technical Progress Reports No. 1, No.2, and No. 3 for FDA Contract No. 223-75-7015, (July 1, 1975March 31 , 1976). Distribution and Clearance of Polybromlnated Biphenyls by Cows. ~Abstract. Journa"Fof ' 'Dalry Science, 551757, (1575). "Distribution and Clearance of Polybrominated Biphenyls By Cows." Submitted for publication to Journal of Dairy Science, (1975 ' . levels of Polychlorinated Biphenyls in ' Adipose fTssuse oF the'General' Population of the Nation. EnvironmentaT "Realth ... Perspectives, 1:79-81, (April, 1972). Epidemiological Study on Yusho Jabies Born to HotEers wholfad ConsumecT'OTl ContininaTed by PCB. Fukuoka Igaku ZassfvT, 5571, 74-80, (1974). Translation by NIH. (Included in MDPH's P3B in Human Breast Milk: Batkground Material) * MONS 056547 ADDENDUM - PBB-PCB BIBLIOGRAPHY Bahn, Anita K., et al. ' Community Nutrition Institute Robertson, Larry W, Melanoma After Exposure to PCB's. The New England JourhaTof Medicine:' 450, (August 19, 1976), "Studies Implicate Fat, PCBs in Human Cancer." Communlty Nutrition Institute Weekly Report VI, 38:7, (September 23, 1976). "PBB: Animal Health Effects." Senate Comnlttee on Health, Social Services MONS 0565-48 SECOND ADDENDUM TO PBB-PCB BIBLIOGRAPHY Doguchi, M. and Fukano, S. Dunckel, Allen E. "Accident Affords Chance to Study Human Toxicology". Hedicaljjews, JAMA, Volume 236 Number 14: 1550^151)1, (October A, 1976). Residue Levels of Polychlorinated Tcrphenyls. Polychlorinated Biphenyls and ODT in Human HI?0-*!. Bulletin of Environmental Contamination antT Toxicology. Vol. 13, Number 1 : 57-63, (197a An Updatinq on the Polybrominated Biphenvl Disaster in Kichiqan. JAYmA. Volume 167. Number' 9`: 838-841, (November 1 , 1975). "Final Report of the Subcommittee on the Health Effects of Polychlorinated Biphenyls and Polybrominated Biphenyls." Department of Heal: Education and Welfare, (July, 1976). Grant, Donald L., et al. PCB Residues in Human Adipose Tissue and Milk. National Conference on Polychlorinated Biphenyl Chicago, (November 19*21, 1975). Hesselberg, R. and Scherr, D. PCB's and p,p' DDE in the Blood of Cachectic Fat'derit's. BuTTeTirTof En'vironrnentaP Contamination and Toxicology. Volume 11, Number 3: 202-205, (1974). Oellnek, Charles F., and Cornellussen, P.E. Kutz, F.W., and Strassman, S.C. Masuada, Yoshlto, et al. Levels of PCB's in the U.S. Food Supply. National Conference on Polychlorinated Biphenyl Chicago, (November 19-21, 1975). Residues of Polychlorinated Biphenyls in the General 'opulation of the United States. National Conference on Polychlorinated Biphenyl Chicago, (November 19-2), 1975). Comparison of Polychlorinated Biphenv1s in Vusho'T'aTents and Ordinary Persons.^'duTfotin of Environmental Contamination and Toxicology. Volume li, Number 3: 213-216, (1974). ' HONS 0565*9 SECOND ADDENDUM TO PBB-PCB BIBLIOGRAPHY McGill, A.E.J ., et. al. Hus 1 a1 C.J., et. al Methods of Estimation Dietary Exposure of the Gener'aT~Populatr6n~ to' Organochlorine Inre'cTfci < Res [dues: "Diet TEnalyses'1965-67. OournYTofTi AOftC. Tolume 55, Number 6: 1245-1258, (1972). Presence of PC8, ODE and DOT in Human Milk in "Nova Scotia . Canada. Bulletin of Envi ronmcntaTTontaroinatk and Toxicology. Volume 12, Number 3: 258-207, (1974). Nlsbct, Ian C .T. Ramachandram, M., et. al. Environmental Transport and Occurrence of PCB'r In 1975. National Conference on Polychlorinated 8iphanyls, Chicago, (November 19-21, 1975). DDT and its Metabolites in Human Body Eat in Tncfia, TIorld Heafth BulTetTn. Volume-'49": ?T7L?38, (1973). Savage, E.P., et. al. . Siyali, D.S., and Strieker, P. Pestjcities in People. Pesticides Monitoring Journal-. "VoTuiii/TTiumber 1: 1-5, (June, 1973) "Erroneous Blood Pesticide Levels Due to Sample Containers with PCBs". The Medical Journal of Australia: 832-833, (October 277197;.') Trout, Paul E The View of the Paper Industry on the Occurrenr of PCB's in the Environment and the Need for Regulation, National Conference on Polychlori.r Biphenyls, Chicago, (November 19-21, 1975), Wassermann, M , , et. al. ' Storage__of Organochlorine Insecticides in Adjn.- Tissue of"Ugandans'! tTulTetin of EnviioiiitentaT `Contamination "ancTToxicology. Volume 12, Numb. 4:501-508, (1974). . West, Robert 1 ., et. a). Hexa_h 1 or whonejConcentratiojns_Jn_H up-i i n_ Mi 1 k. "BuTi etin "of Envi rorTmantaYTontaminatTc n "and Toxicolog/, Volume 13, Number 2: 16/-169, (19 HONS 056550 Ootober 11, 1937 Experimental -orfc In animals shows that prolonged exposure to Aroolor vapors e/ volved at high temperatures or by reseated oral Ingestion will lead to systemlo toxlo effeots. Repeated bodily contaot with the liquid ri Aroolors may lead to an none-fona skin eruption. Suitable draft ventilation to control the vapors evolved at elevated temperatures, ~ as well as proteotlon by suitable garments " from extensive bodily oontaot with the liq uid Aroolors, should prevent any untoward ^effeet." In talking with Dr. Kelly before theee three paragraphs were written, we agreed that they might as well be phrased so that they oould be used not only In the Aroolor booklet, but quoted In correspondence as that may be neoessary. L.A. tfatt ( ' Attachment 3-2 > HONS 056551 PCB POISONING AND POLLUTION B4iui bj> JCentaro Hiouchi Fukuoka University Hospital, Japan 1976 8 KODANSHA LTD. Tokyo ACADEMIC PRESS Now York Son FroncUco London A Svbildigpy of Mortout Iroct Jo*OF>victi, Pvblitht'i HONS 056552 KODANSHA SCIENTIFIC BOOKS Copyright 0 1976 by Kodantha Ltd. All right* reaerved No pan of thb book may be reproduced in any form, by photoitat. microfilm, retrieval syitem, or any other mean*, without the written permiMion of Kodaruh* Ltd. (except In theca* of brief quotation for critkbm or review) I.S.B.N. CM2-M7WO-2 Library of Congee# Catalog Card Number 76*241M Cf-^iiuW i; KODANSHA LTD. 12*21, Otowa 2*chome, Bunkyo-ku, Tokyo 112, Japan tnd ACADEMIC PRESS, INC. ID Fifth Avenue. New York, N.Y. 10003 ACADEMIC PRESS, (LONDON) LTD. 24/28 Oval Road, London NW I PlONTlt) IN JaFAN HONS 056553 Fig. 7.15 Swelliti| of the eyelid* and browniih akin pigmentation of a new-born baby. Fig. 7.9 Nail pigmentation ($. K,, aged Fig. 7.10 Pigmentation of the palpebral 5, female). conjunctiva and timbal conjunctiva. Fig. 7,|| Oingisal pigmentation (N. K., F*g. 7.li I'igmcntaiion uf die lips, aged 36, female). HONS 05655<i Contributors Kentaro Hiouchi, Fukuoka University Hospital, Nishi-ku, Fukuoka 814, Japan Chistto Hirayama, Third Department of Internal Medicine, Faculty of Medicine, Kyushu University, Fukuoka 812, Japan. MaiaMro Kikvcni, Department of Pathology, Faculty of Medicine, Fukuoka University, Fukuoka 814, Japan Hiromu Koda, Department of Dermatology, Faculty of Medicine, Kyushu University, Fukuoka 812, Japan Masanori Kuratsuni, Department of Public Heath, Faculty of Medicine, Kyushu University, Fukuoka 812, Japan Yosbito Masuda, Dailcht College of Pharmaceutical Sciences, Fukuoka 815, Japan Hiroya Tanam, Food Hygiene Laboratory, Sagami Women's University, Sagamlhart-ihl 228, Japan Roy Tatsukawa, College of Agriculture, Ehime University, Matsuyama* shi 780, Japan KHchi Ueda, Laboratory of Hygiene and Oral Health, Showa University, Tokyo 142, Japan Harukuni Uram, Department of Dermatology, Faculty of Medicine, Kyushu University, Fukuoka 812, Japan Shin'ichi Yoshmara, Faculty of Pharmaceutical Sciences, Kyushu Uni* versity, Fukuoka 812, Japan Hidetoshi Yoshimura, Faculty of Pharmaceutical Sciences, Kyushu Uni versity, Fukuoka 812, Japan HONS 056555 Preface Recently, the various health hazards and dangers of indiscriminate discharge of industrial wastes into the environment have been emphasis* td, Also, strong warnings have been issued, both officially and unoffi cially, against polluting the environment with toxic or otherwise harmful Industrial products. However, It Is distressing that, so often, severe cases of poisoning are required before the dangers are fully recognized or effec tive countermeasures can be implemented or enforced. The present book coven the broad field of PCB poisoning and pollu tion In Japan. It emphasized the case ofdirect human poisoning by oral Intake of PCB-contaminated rice-oil (Yusho), and also includes sections on the experimental and toxicological aspects of PCB poisoning, and on the general pollution of the natural environment by PCB's. The medical description of Yusho covers epidemiologic, pathologic, clinical and therapeutic studies, and there are detailed descriptions of the dermal symptoms (which resemble chloracne) and other major clinical manifestations of the disease, in both adults and children. The sections on toxicology cover various studies, mostly animal experiments, on the physiological and other effects of PCB's, and on the absorption, distribu tion and metabolism of PCB's In the animal body. The sections devoted to environmental pollution In Japan describe both the mlcroanalytlcal methodology for PCB's, as adopted in Japan, and also the extent of pollu tion of the atmosphere, freshwater and marine environments, soil, agricul tural products, fish, birds, and man. As editor for this project, I wish to express my appreciation to all authors for their contributions, and would also like to take this opportu nity to thank the staff of Kodaniht for their editorial and linguistic assist ance in the preparation of the English manuscripts comprising this book. July, 1976 ' K. Hiouchi HONS 056556 Contents Contributor!, v Prefect, vii PART t: PCD POKONINO Chapter I. Outline, 9 Chapter 2. Epidemiologic Studies on Yusho, 9 2.1. Introduction, 9 2.2. Clinical Symptoms, 10 2.$. Descriptive Epidemiologic Studies, 11 2.4. Analytical Epidemiologic Studies, 13 2.3. Dose-Response Relationship, 13 2.6. Toxic Agent, 16 2.7. Amount of Kanechlor 400 Ingested by Patients, 16 2.9. Babies Born to Patients and to N'on-affetted Wives of Patients, and Babies Breast-fed by Patients, 16 2.9. Growth of Yuiho Children, 22 2.10. PCB's in Yusho Patients, 22 2.11. Deaths among Yusho Patients, 22 Chapter 3. Toxicological Aspects 1: the Toxicity of PCB's, 25 3.1. Introduction, 23 3.2. Acute Toxicity for Experimental Animats, 26 3.3. Subacute and Chronic Studies, 26 3.4. Biochemical Reactions, 26 3.3. Endocrine Effects, 31 3.6. Immunosuppressive Activity, 32 3.7, Carcinogenesis, 32 3.6. Effects on Successive Generations, 34 HONS 05655? 3.9. Toxicological Significance of Coexistent Byproducts of Hislier Toxicity. S3 ' 3.10. Comparative Toxicity of Synthetic Isomers, 36 3.11. Possible Health Effects: Present and Future, 38 Chapter 4. Toxicological Aspects II: the Metabolic Fate of PCB s and Their Toxicological Evaluation, 41 4.1. Introduction, 41 4.2. Absorption, Distribution and Elimination of PCB's, 42 4.9. Metabolism of PCB's, 49 4.4. Toxicological Evaluation of the Metabolism of PCB's, 62 Chapter 5. The Pathology of Yusho, 69 5.1. Introduction, 69 3.2> Autopsy Findings of Vusho''adents, 70 5.3. Cutaneous Changes In Yusho Patients, 77 5.4. Cai-chromatographic Analysis of the PCB Content of Visceral Organs in Yusho Patients, 78 $.3. Discussion, 82 Chapter 6. Clinical Aspects of PCB Poisoning, B7 6.1. Introduetion, 87 6.2. Clinical Manifestations, 88 6.3. Effects on Fetal and Infant Life, 6.4. Laboratory Findings, 100 6.5. Treatment, 102 99 Chapter 7. The Dermal Symptomatology of Yusho, 7.1. Introduetion, 105 7.2. Clinical Symptoms, 106 7.3. Treatment, 118 7.4. Clinical Course, 120 7.3. Mechanisms, 121 7.6. Conclusion, 122 ' 105 PART ti: PCB POLLUTION Chapters. PCB Microanalysis, 127 8.1. Apparatus, 127 8.2. Reagents, 127 8.3. Standards, 128 ' 6.4. Preparation of Sample Solution, 8.5. Measurement of PCB's, 132 128 Chapter 9. PCB Pollution of the Japanese Environment, 9.1. Introduction, 14? 9.2. Production, Use and Properties of PCB's 146 9.3. Environmental PCB Pollution in Japan, 153 Subject Index, 181 147 HONS 056556