Benzyl Acetate
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NATIONAL TOXICOLOGY PROGRAM Technical Report Series No. 431 TOXICOLOGY AND CARCINOGENESIS STUDIES OF BENZYL ACETATE (CAS NO. 140-11-4) IN F344/N RATS AND B6C3F, MICE (FEED STUDIES) U.S.DEPARTMENT OF HEALTHANDHUMANSERVICES PublicHealthService 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 HumanServices (DHHS): the National Cancer Institute (NCI), National Institutesof Health; the National Institute of Environmental Health Sciences (NIEHS), National Institutes of Health; the Nation(a1 Center for 'Toxicological Research (NCTR),Food and Drug Administration; and the National Institute for Occupational Safety and Health (NIOSH), Centersfor 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 andweire conducted in compliance with NTP Iaboratory health and safety requirements and must meet or exceed ,all applicable federal, state, andlocal health andsafety regulations. Animal care anduse were in accordance with the Public Health Service Policy on Humane Care and Useof Animals. The prechronic and chronic studies wereconductedincompliancewith Food andDrugAdministration (FDA) Good LaboratoryPractice 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 inlaboratoryanimals (usually two species,rats and mice). Chemicals selected for NTF' toxicology and carcinogenesis studies are chosen primarily on the bases of human exposure, level of production, and chemical structure. Selection per se is not an indicator of a chemical's carcinogenic potential. TheseNTPTechnicalReportsare available for salefrom the NationalTechnicalInformation Service, U.S. Department of Commerce, 5285 Port Royal Road, Springfield, VA 22161 (703-487-4650). Single copies of this Technical Report are available without charge while supplies last from Central Data Management, NIEHS, P.,O. Box 12233, MD AO-01, Research Triangle Park, NC 27709 (919-541-1371). NTP TECHNICAL REPORT ON THE TOXICOLOGY AND CARCINOGENESIS STUDIES OF BENZYL ACETATE (CAS NO. 140-11-4) IN F344/N RATS AND B6C3Fl MICE (FEED STUDIES) NATIONAL TOXICOLOGY PROGRAM P.O. Box 12233 Research Triangle Park, NC 27709 September 1993 NTP TR 431 NIH Publication No. 93-3162 U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES Public Health Service National Institutes of Health 2 Benzyl Acetate, NTP TR 431 CONTRIBUTORS National Toxicology Program NTP Pathology Working Group Evobroledondinterpreteddandrtynmed@a%p Evaluated slides,prepared pathologyreport on rats (29 May 1991) KM. Abdo, Ph.D. C.J. Alden, PbD. T.M. Monticello, D.V.M., Ph.D., Chair Pathology Associates, Inc. G.A. Boorman, D.V.M., Ph.D. M.A. Bryant, D.V.M. D.A. Bridge, B.S. U.S. Army Medical Reserves Instructor S.L. Eustis, D.V.M., PLD. B.F. Hamilton, D.V.M., Ph.D. T.J. Goehl, Ph.D. Experimental Pathology Laboratories, Inc. R.A. Griesemer, D.V.M., Ph.D. M.P. Jokinen, D.V.M. J.R. Hailey, D.V.M. National Toxicology Program J.K Haseman, Ph.D. M.M. McDonald, D.V.M., PbD. G.N. Rao, D.V.M., PbD. National Toxicology Program C.C. Shackelford, D.V.M., M.S., Ph.D. S. Qureshi, B.V.Sc., PbD. B.A. Schwetz, D.V.M., Ph.D. Sandoz, Ltd. D.B. Walt.ers, PbD. C.C. Shackelford, D.V.M., M.S., Ph.D. National Toxicology Program KL. Witt, M.S., Oak Ridge Associated Universities A. Steber (observer) University of Wisconsin Southern Research Institute Conducted studies, evaluated pathologywgs Evaluated slih,prepared pathologyrepon on mice (15 Augurt 1991) J.D. Prejean, Ph.D., Principal Investigator D.R. Farnell, D.V.M., P&D. PK Hildebrandt, D.V.M., Chair H.D. Gila, D.V.M., PbD. PATHCO,Inc. J.S. Heath, D.V.M. S. Botts, M.S., D.V.M. Experimental Pathology Laboratories, Inc. M.P. Jokinen, D.V.M. Experimental Pathology Laboratories, Inc. National Toxicology Program Providkdpathologyquality asncrmrce J.R. Hailey, D.V.M. National Toxicology Program J.F. Hardisty, D.V.M., principal Investigator M.M. McDonald, D.V.M., Ph.D. S. Botts, M.S., D.V.M. National Toxicology Program B.F. Hamilton, D.V.M., Ph.D. KT.Morgan, B.V.Sc., M.R.C.V.S., Ph.D. Chemical Industry Institute of Toxicology Dynamac Corporation E. Sander, M.D. (observer) Prepmd quality assurance ma% Bayer AG C.C. Shackelford, D.V.M., M.S., Ph.D. S. Brecher, Ph.D., PrincipalInvestigator National Toxicology Program Biotechnical Services, Inc. hepared Technical Repon D.D. Lambright, Ph.D., PrincipalInvestigator G.F.Corley, D.V.M. P.S. Keightley, B.k T.A. King-Hunter, B.S.H.S. H.S. Lindsay, B.A. 3 CONTENTS ABSTRACT ................................................................... 5 EXPLANATION OF LEVELS OF EVIDENCE OF CARCINOGENIC ACTIVITY ............... 9 TECHNICAL REPORTS REVIEW SUBCOMMITTEE ................................... 10 SUMMARY OF TECHNICAL REPORTS REVIEW SUBCOMMITTEE COMMENTS ........... 11 INTRODUCTION .............................................................. 13 MATERIALSANDMETHODS .................................................... 19 RESULTS .................................................................... 27 DISCUSSION AND CONCLUSIONS ................................................ 49 REFERENCES ................................................................ 53 APPENDIX A Summary of Lesions in Male Rats in the 2-Year Feed Study ofBenylAcetate .................................................. 59 APPENDIX B Summary of Lesions in Female Rats in the 2-Year Feed Study ofBenzylAcetate .................................................. 101 APPENDIXC Summary of Lesions in Male Mice in the 2-Year Feed Study ofBenzylAcetate .................................................. 139 APPENDIX D Summary of Lesions in FemaleMicein the 2-Year Feed Study ofBenylAcetate .................................................. 183 APPENDIX E Genetic Toxicology ................................................. 225 APPENDIX F Organ Weights and Organ-Weight-to-Bodyweight Ratios .................... 241 APPENDIXG Hematology and Clinical Chemistry Results ............................... 249 APPENDIXHPancreatic EnzymeLevels ............................................ 255 APPENDIXI Chemical Characterization and Dose Formulation Studies ................... 259 APPENDIX J Feed and Compound Consumption in the 2-YearFeed Studies ................ 273 APPENDIX K Ingredients. Nutrient Composition. andContaminant Levels in NIH-07 Rat and Mouse Ration ..................................... 279 APPENDIX L Sentinel Animal Program ............................................ 285 ABSTRACT BENZYL ACETATE CAS NO. 140-114 ChemicalFormula: CJ-I,,O, Molecular Weight: 150.17 Gynongms: acetic add benyl ester,acetic acid phenyl methyl ester,(acetoxymethyl)benzene,acemaytoluene, benzyl ethanoate, phenylmethyl acetate Benzyl acetate is used as a flavoring agent in foods, 50,OOO ppm (0, 230, 460, 900, 1,750,or 3,900 mg/kg as a fragrance in soaps and perfumes, as a solvent for body weightfor males and 0,240,480,930,1,870,or cellulose acetate and nitrate, and as a component of 4,500 mgkg for females) benzylacetate for 13 weeks. printing inks and varnish removers. The NTP previ- Nine male and nine female rats receiving 50,OOO ppm ously studied the toxicology and carcinogenicity of benzyl acetate died or were killed moribund between this chemical in F344/Nrats and B6C3F, mice using weeks 2and 8 of the study. The mean body weight the gavage route of administration and corn oilas a gain and the final mean body weight of 25,OOO ppm vehicle.Benzyl acetate increased the incidences of males were significantlylower (P10.01)than those of pancreatic acinar cell adenomas in male rats and the the control group. Feed consumption by exposed incidences of hepatocellular adenomas and fore- rats, except the 25,OOO and 50,OOO ppm males and stomach neoplasms in male and female mice. 50,OOO ppm females, was similar to that by the Because of the confounding effect of corn oil on the controls. The reduced feed consumption by 25,OOO incidences of pancreatic neoplasms and because of and 50,OOO ppm males and 50,OOO ppm females may controversy over the use of the gavage route of have been due to toxicity or decreased palatability. administration, the NTP decided to restudy benzyl Tremors and ataxia occurred only in the 50,OOO ppm acetate using the dosed feed route of administration. rats. These findings were first observed on day 15 in In these repeat studies, male and female F344/Nrats nine males and six females and continued until the and B6C3Fl mice received benzyl acetate (at least end of the study. Cholesterol levels in 1v00 and 98% pure) in feed for 13weeks and 2years. Genetic 25,OOO ppmfemales and triglyceride levels in toxicology studies were conducted in Salmonella 25,OOO ppmfemales were lower than those in the lyphimurium, cultured Chinese hamster ovary cells, controls.