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T-Butyl Alcohol National Toxicology Program Toxicity Report Series Number 53 NTP Technical Report on Toxicity Studies of t-Butyl Alcohol (CAS No. 75-65-0) Administered by Inhalation to F344/N Rats and B6C3F1 Mice Joel Mahler, D.V.M., Study Scientist National Toxicology Program Post Office Box 12233 Research Triangle Park, NC 27709 NIH Publication 97-3942 July 1997 United States Department of Health and Human Services Public Health Service National Institutes of Health Note to the Reader The National Toxicology Program (NTP) is made up of four charter agencies of the United States Department of Health and Human Services (DHHS): • the National Cancer Institute (NCI) of the National Institutes of Health; • the National Institute of Environmental Health Sciences (NIEHS) of the National Institutes of Health; • the National Center for Toxicological Research (NCTR) of the Food and Drug Administration; and • the National Institute for Occupational Safety and Health (NIOSH) of the Centers for Disease Control. In July 1981, the Carcinogenesis Bioassay Testing Program was transferred from NCI to NIEHS. NTP coordinates the relevant Public Health Service programs, staff, and resources that are concerned with basic and applied research and with biological assay development and validation. 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. NTP designs and conducts studies to characterize and evaluate the toxicologic potential of selected chemicals in laboratory animals (usually two species, rats and mice). Chemicals selected for NTP toxicology studies are chosen primarily on the bases of human exposure, level of production, and chemical structure. The interpretive conclusions presented in this Toxicity 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 toxic potential. The studies described in this toxicity study report were performed under the direction of NIEHS and were conducted in compliance with NTP laboratory health and safety requirements. These studies met or exceeded all applicable federal, state, and local health and safety regulations. Animal care and use were in accord and compliance with the Public Health Service Policy on Humane Care and Use of Animals. Single copies of this report are available without charge, while supplies last, from the NTP Central Data Management (telephone number 919/541-3419). NTP Central Data Management MD E1-02 NIEHS Post Office Box 12233 Research Triangle Park, NC 27709 National Toxicology Program Toxicity Report Series Number 53 NTP Technical Report on Toxicity Studies of t-Butyl Alcohol (CAS No. 75-65-0) Administered by Inhalation to F344/N Rats and B6C3F1 Mice Joel Mahler, D.V.M., Study Scientist National Toxicology Program Post Office Box 12233 Research Triangle Park, NC 27709 United States Department of Health and Human Services Public Health Service National Institutes of Health 2 t-Butyl Alcohol, NTP TOX 53 CONTRIBUTORS This NTP report on the toxicity studies of t-butyl alcohol is based primarily on 18-day and 13-week studies that took place from August 1986 through June 1987. National Toxicology Program Experimental Pathology Laboratories, Inc. Evaluated experiment, interpreted results, and reported findings Provided pathology quality assessment Joel Mahler, D.V.M., Study Scientist H. Roger Brown, D.V.M. John R. Bucher, Ph.D. Robert E. Chapin, Ph.D. Michael R. Elwell, D.V.M., Ph.D. Environmental Health Research and Thomas J. Goehl, Ph.D. Testing, Inc. Joseph K. Haseman, Ph.D. Provided sperm morphology and vaginal cytology evaluation Ghanta N. Rao, D.V.M., Ph.D. Teresa Cocanougher, B.A. Gregory S. Travlos, D.V.M. Dushant K. Gulati, Ph.D. Kristine L. Witt, M.S. Susan Russell, B.A. Oak Ridge Associated Universities Battelle Columbus Laboratories Analytical Sciences, Inc. Provided statistical analyses Principal contributors Richard Morris, M.S., Principal Investigator Arthur C. Peters, D.V.M., Principal Investigator Patricia M. Athey Sarah R. Lloyd, M.S. Douglas K. Craig, Ph.D. Nicole G. Mintz, B.S. Sondra L. Grumbein, D.V.M., Ph.D. Milton R. Hejtmancik, Ph.D. Biotechnical Services, Inc. Provided toxicity report preparation NTP Pathology Review Susan R. Gunnels, M.A., Principal Investigator Evaluated slides and prepared pathology report Gloria Gordon, M.A. John Seely, D.V.M. Lynn M. Harper, B.S. PATHCO, Inc. Michael L. Rainer, B.S. Michael R. Elwell, D.V.M., Ph.D. Waynette D. Sharp, B.A., B.S. NIEHS t-Butyl Alcohol, NTP TOX 53 3 PEER REVIEW The draft report on the toxicity studies of t-butyl alcohol was evaluated by the reviewers listed below. These reviewers serve as independent scientists, not as representatives of any institution, company, or governmental agency. In this capacity, reviewers determine if the design and conditions of these NTP studies are appropriate and ensure that the toxicity study report presents the experimental results and conclusions fully and clearly. James Bond, Ph.D. Gary P. Carlson, Ph.D. Chemical Industry Institute of Toxicology School of Health Sciences Research Triangle Park, NC Purdue University West Lafayette, IN 4 t-Butyl Alcohol, NTP TOX 53 CONTENTS ABSTRACT .............................................................. 5 INTRODUCTION ........................................................... 9 Physical Properties, Production, Use, and Human Exposure ............................ 9 Absorption, Distribution, Metabolism, and Excretion................................. 10 Toxicity .............................................................. 12 Reproductive and Developmental Toxicity ........................................ 14 Carcinogenicity ......................................................... 15 Genetic Toxicity ........................................................ 16 Study Rationale and Design ................................................. 16 MATERIALS AND METHODS ................................................... 17 Procurement and Characterization of t-Butyl Alcohol ................................. 17 Vapor Generation System .................................................. 17 Concentration Monitoring .................................................. 18 Chamber Characterization .................................................. 18 Toxicity Study Designs .................................................... 20 Statistical Methods ....................................................... 27 Genetic Toxicity Studies ................................................... 27 Quality Assurance Methods ................................................. 31 RESULTS ............................................................... 33 18-Day Inhalation Study in F344/N Rats ......................................... 33 13-Week Inhalation Study in F344/N Rats ........................................ 35 18-Day Inhalation Study in B6C3F1 Mice ........................................ 41 13-Week Inhalation Study in B6C3F1 Mice ....................................... 43 Genetic Toxicology....................................................... 46 DISCUSSION ............................................................. 47 REFERENCES ............................................................ 49 APPENDIX A Organ Weights and Organ-Weight-to-Body-Weight Ratios .................. A-1 APPENDIX B Hematology, Clinical Chemistry, and Urinalysis Results ................... B-1 APPENDIX C Reproductive Tissue Evaluations and Estrous Cycle Characterization .......... C-1 APPENDIX D Genetic Toxicology............................................. D-1 5 ABSTRACT t-BUTYL ALCOHOL CAS No. 75-65-0 Chemical Formula: C4H10O Molecular Weight: 74.12 Synonyms: 2-Methyl-2-propanol, 2-methylpropan-2-ol, TBA, t-butanol, t-butyl hydroxide, trimethyl carbinol, trimethyl methanol t-Butyl alcohol is widely used in the manufacture of perfumes and a variety of cosmetics. It is also used as a raw material in the production of isobutylene, which may be used to produce methyl tertiary butyl ether, a common gasoline additive, or to produce butyl elastomers used in the production of automobile tires. The National Cancer Institute nominated t-butyl alcohol to the NTP for study as a result of a review of chemicals found in drinking water. In addition to the high annual production and the potential for occupational exposure, there is also a potential for human exposure to t-butyl alcohol by the inhalation route from its use as an additive in unleaded gasoline. Therefore, toxicity studies of t-butyl alcohol were conducted in male and female F344/N rats and B6C3F1 mice by whole-body inhalation. Animals were evaluated for hematology, clinical chemistry, urinalysis, reproductive toxicity, and histopathology. The genetic toxicity of t-butyl alcohol was assessed by testing the ability of the chemical to induce mutations in various strains of Salmonella typhimurium and L5178Y mouse lymphoma cells or sister chromatid exchanges and chromosomal aberrations in cultured Chinese hamster ovary cells, and by measuring the frequency of micronucleated erythrocytes in rat bone marrow and mouse peripheral blood. 6 t-Butyl Alcohol, NTP TOX 53 In the 18-day inhalation studies, groups of five male and five female rats and mice were exposed to t-butyl alcohol by
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