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TR-466: Ethylbenzene (CASRN 100-41-4) in F344/N Rats And

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TR-466: Ethylbenzene (CASRN 100-41-4) in F344/N Rats And NTP TECHNICAL REPORT ON THE TOXICOLOGY AND CARCINOGENESIS STUDIES OF ETHYLBENZENE (CAS NO. 100-41-4) IN F344/N RATS AND B6C3F1 MICE (INHALATION STUDIES) NATIONAL TOXICOLOGY PROGRAM P.O. Box 12233 Research Triangle Park, NC 27709 January 1999 NTP TR 466 NIH Publication No. 99-3956 U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES Public Health Service National Institutes of Health FOREWORD The National Toxicology Program (NTP) is made up of four charter agencies of the U.S. Department of Health and Human Services (DHHS): the National Cancer Institute (NCI), National Institutes of Health; the National Institute of Environmental Health Sciences (NIEHS), National Institutes of Health; the National Center for Toxicological Research (NCTR), Food and Drug Administration; and the National Institute for Occupational Safety and Health (NIOSH), Centers for Disease Control. In July 1981, the Carcinogenesis Bioassay Testing Program, NCI, was transferred to the NIEHS. The NTP coordinates the relevant programs, staff, and resources from these Public Health Service agencies relating to basic and applied research and to biological assay development and validation. The NTP develops, evaluates, and disseminates scientific information about potentially toxic and hazardous chemicals. This knowledge is used for protecting the health of the American people and for the primary prevention of disease. The studies described in this Technical Report were performed under the direction of the NIEHS and were conducted in compliance with NTP laboratory health and safety requirements and must meet or exceed all applicable federal, state, and local health and safety regulations. Animal care and use were in accordance with the Public Health Service Policy on Humane Care and Use of Animals. The prechronic and chronic studies were conducted in compliance with Food and Drug Administration (FDA) Good Laboratory Practice Regulations, and all aspects of the chronic studies were subjected to retrospective quality assurance audits before being presented for public review. These studies are designed and conducted to characterize and evaluate the toxicologic potential, including carcinogenic activity, of selected chemicals in laboratory animals (usually two species, rats and mice). Chemicals selected for NTP toxicology and carcinogenesis studies are chosen primarily on the bases of human exposure, level of production, and chemical structure. The interpretive conclusions presented in this Technical Report are based only on the results of these NTP studies. Extrapolation of these results to other species and quantitative risk analyses for humans require wider analyses beyond the purview of these studies. Selection per se is not an indicator of a chemical’s carcinogenic potential. Listings of all published NTP reports and ongoing studies are available from NTP Central Data Management, NIEHS, P.O. Box 12233, MD E1-02, Research Triangle Park, NC 27709 (919-541-3419). The Abstracts and other study information for 2-year studies are also available at the NTP’s World Wide Web site: http://ntp-server.niehs.nih.gov. NTP TECHNICAL REPORT ON THE TOXICOLOGY AND CARCINOGENESIS STUDIES OF ETHYLBENZENE (CAS NO. 100-41-4) IN F344/N RATS AND B6C3F1 MICE (INHALATION STUDIES) NATIONAL TOXICOLOGY PROGRAM P.O. Box 12233 Research Triangle Park, NC 27709 January 1999 NTP TR 466 NIH Publication No. 99-3956 U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES Public Health Service National Institutes of Health These studies were supported in part by funds from the Comprehensive Environmental Response, Compensation, and Liability Act trust fund (Superfund) by an interagency agreement with the Agency for Toxic Substances and Disease Registry, U.S. Public Health Service. 2 Ethylbenzene, NTP TR 466 CONTRIBUTORS National Toxicology Program NTP Pathology Working Group Evaluated and interpreted results and reported findings Evaluated slides, prepared pathology report on rats (13 July 1995) P.C. Chan, Ph.D., Study Scientist D.A. Bridge, B.S. J.C. Seely, D.V.M., Chairperson J.R. Bucher, Ph.D. PATHCO, Inc. N. Barlow, Ph.D. R.E. Chapin, Ph.D. North Carolina State University J.R. Hailey, D.V.M. D. Dixon, D.V.M., Ph.D. J.K. Haseman, Ph.D. National Toxicology Program J. Mahler, D.V.M. J.R. Hailey, D.V.M. R.R. Maronpot, D.V.M. National Toxicology Program G.N. Rao, D.V.M., Ph.D. R.A. Herbert, D.V.M., Ph.D. J.H. Roycroft, Ph.D. National Toxicology Program A. Radovsky, D.V.M., Ph.D. C.S. Smith, Ph.D. National Toxicology Program G.S. Travlos, D.V.M. C.C. Shackelford, D.V.M., M.S., Ph.D. D.B. Walters, Ph.D. Experimental Pathology Laboratories, Inc. K.L. Witt, M.S., Oak Ridge Associated Universities M. Torii, D.V.M., Ph.D. National Toxicology Program D. Wolf, D.V.M., Ph.D. IIT Research Institute Chemical Industry Institute of Toxicology Conducted 2-year studies, evaluated pathology findings Evaluated slides, prepared pathology report on mice (12 December 1995) C. Aranyi, M.S., Principal Investigator D.T. Kirkpatrick, Ph.D. J.C. Seely, D.V.M., Chairperson A. Snelson, Ph.D. PATHCO, Inc. S. Asanoi, Ph.D., Observer National Toxicology Program G.A. Boorman, D.V.M., Ph.D. Experimental Pathology Laboratories, Inc. National Toxicology Program Provided pathology quality assurance R. Cattley, D.V.M., Ph.D. Chemical Industry Institute of Toxicology J.F. Hardisty, D.V.M., Principal Investigator J. Hellman, D.V.M. C.C. Shackelford, D.V.M., M.S., Ph.D. National Toxicology Program R.A. Herbert, D.V.M., Ph.D. National Toxicology Program Dynamac Corporation A. Radovsky, D.V.M., Ph.D. Prepared quality assurance audits National Toxicology Program C.C. Shackelford, D.V.M., M.S., Ph.D. S. Brecher, Ph.D., Principal Investigator Experimental Pathology Laboratories, Inc. R.C. Sills, D.V.M., Ph.D. National Toxicology Program Ethylbenzene, NTP TR 466 3 Analytical Sciences, Inc. Biotechnical Services, Inc. Provided statistical analyses Prepared Technical Report R.W. Morris, M.S., Principal Investigator S.R. Gunnels, M.A., Principal Investigator S.R. Lloyd, M.S. L.M. Harper, B.S. N.G. Mintz, B.S. A.M. Macri-Hanson, M.A., M.F.A. E.S. Rathman, M.S. 4 CONTENTS ABSTRACT ............................................................. 5 EXPLANATION OF LEVELS OF EVIDENCE OF CARCINOGENIC ACTIVITY ............. 10 TECHNICAL REPORTS REVIEW SUBCOMMITTEE ................................ 11 SUMMARY OF TECHNICAL REPORTS REVIEW SUBCOMMITTEE COMMENTS .......... 12 INTRODUCTION ......................................................... 15 MATERIALS AND METHODS ................................................ 21 RESULTS ............................................................... 29 DISCUSSION AND CONCLUSIONS ............................................ 47 REFERENCES ........................................................... 51 APPENDIX A Summary of Lesions in Male Rats in the 2-Year Inhalation Study of Ethylbenzene ................................................ 57 APPENDIX B Summary of Lesions in Female Rats in the 2-Year Inhalation Study of Ethylbenzene ................................................ 99 APPENDIX C Summary of Lesions in Male Mice in the 2-Year Inhalation Study of Ethylbenzene ................................................ 131 APPENDIX D Summary of Lesions in Female Mice in the 2-Year Inhalation Study of Ethylbenzene ................................................ 167 APPENDIX E Genetic Toxicology ............................................. 201 APPENDIX F Chemical Characterization and Generation of Chamber Concentrations ......... 211 APPENDIX G Ingredients, Nutrient Composition, and Contaminant Levels in NIH-07 Rat and Mouse Ration ................................... 221 APPENDIX H Sentinel Animal Program ......................................... 225 5 ABSTRACT CH2CH3 ETHYLBENZENE CAS No. 100-41-4 Chemical Formula: C8 H10 Molecular Weight: 106.16 Synonyms: EB; ethylbenzol; phenylethane Ethylbenzene is mainly used in the manufacture of 2-YEAR STUDY IN RATS styrene. Ethylbenzene is also a major component of Groups of 50 male and 50 female F344/N rats were mixed xylenes used as solvents in agricultural and exposed to 0, 75, 250, or 750 ppm ethylbenzene by home insecticide sprays, rubber and chemical manu­ inhalation, 6 hours per day, 5 days per week, for facturing, and household degreasers, paints, adhe­ 104 weeks. sives, and rust preventives. Ethylbenzene is also used as an antiknock agent in aviation and motor fuels. Survival and Body Weights Ethylbenzene was nominated for study by the National Survival of male rats in the 750 ppm group was Institute for Occupational Safety and Health (NIOSH) significantly less than that of the chamber controls. and the Occupational Safety and Health Admin­ Mean body weights of 250 and 750 ppm males were istration (OSHA) because of its potential for wide­ generally less than those of the chamber controls spread human exposure and because of its structural beginning at week 20. Mean body weights of exposed similarity to benzene and toluene. Male and female groups of females were generally less than those of F344/N rats and B6C3F mice were exposed to 1 chamber controls during the second year of the study. ethylbenzene (greater than 99% pure) by inhalation for 2 years. Genetic toxicology studies were con­ ducted in Salmonella typhimurium, mouse lymphoma Pathology Findings cells, cultured Chinese hamster ovary cells, and In male rats exposed to 750 ppm, the incidences of mouse peripheral blood erythrocytes. In previously renal tubule adenoma and adenoma or carcinoma reported 13-week toxicity studies in which F344/N
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