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TR-541: Formamide (CASRN 75-12-7) in F344/N Rats and B6C3F1 Mice (Gavage Studies)

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TR-541: Formamide (CASRN 75-12-7) in F344/N Rats and B6C3F1 Mice (Gavage Studies) NTP TECHNICAL REPORT ON THE TOXICOLOGY AND CARCINOGENESIS STUDIES OF FORMAMIDE (CAS NO. 75-12-7) IN F344/N RATS AND B6C3F1 MICE (GAVAGE STUDIES) NATIONAL TOXICOLOGY PROGRAM P.O. Box 12233 Research Triangle Park, NC 27709 July 2008 NTP TR 541 NIH Publication No. 08-5884 National Institutes of Health Public Health Service U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES FOREWORD The National Toxicology Program (NTP) is an interagency program within the Public Health Service (PHS) of the Department of Health and Human Services (HHS) and is headquartered at the National Institute of Environmental Health Sciences of the National Institutes of Health (NIEHS/NIH). Three agencies contribute resources to the program: NIEHS/NIH, the National Institute for Occupational Safety and Health of the Centers for Disease Control and Prevention (NIOSH/CDC), and the National Center for Toxicological Research of the Food and Drug Administration (NCTR/FDA). Established in 1978, the NTP is charged with coordinating toxicological testing activities, strengthening the science base in toxicology, developing and validating improved testing methods, and providing information about potentially toxic substances to health regulatory and research agencies, scientific and medical communities, and the public. The Technical Report series began in 1976 with carcinogenesis studies conducted by the National Cancer Institute. In 1981, this bioassay program was transferred to the NTP. The studies described in the Technical Report series are designed and conducted to characterize and evaluate the toxicologic potential, including carcinogenic activity, of selected substances in laboratory animals (usually two species, rats and mice). Substances selected for NTP toxicity and carcinogenicity studies are chosen primarily on the basis of human exposure, level of production, and chemical structure. The interpretive conclusions presented in NTP Technical Reports are based only on the results of these NTP studies. Extrapolation of these results to other species, including characterization of hazards and risks to humans, requires analyses beyond the intent of these reports. Selection per se is not an indicator of a substance’s carcinogenic potential. The NTP conducts its studies in compliance with its laboratory health and safety guidelines and FDA Good Laboratory Practice Regulations and must meet or exceed all applicable federal, state, and local health and safety regulations. Animal care and use are in accordance with the Public Health Service Policy on Humane Care and Use of Animals. Studies are subjected to retrospective quality assurance audits before being presented for public review. NTP Technical Reports are indexed in the NIH/NLM PubMed database and are available free of charge electronically on the NTP website (http://ntp.niehs.nih.gov) or in hardcopy upon request from the NTP Central Data Management group at [email protected] or (919) 541-3419. NTP TECHNICAL REPORT ON THE TOXICOLOGY AND CARCINOGENESIS STUDIES OF FORMAMIDE (CAS NO. 75-12-7) IN F344/N RATS AND B6C3F1 MICE (GAVAGE STUDIES) NATIONAL TOXICOLOGY PROGRAM P.O. Box 12233 Research Triangle Park, NC 27709 July 2008 NTP TR 541 NIH Publication No. 08-5884 National Institutes of Health Public Health Service U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES 2 CONTRIBUTORS National Toxicology Program NTP Pathology Working Group Evaluated and interpreted results and reported findings Evaluated slides and prepared pathology report on rats (June 02, 2005) R.D. Irwin, Ph.D., Study Scientist Chairperson S.A. Elmore, D.V.M., M.S., Study Pathologist S.A. Elmore, D.V.M., M.S., ILS, Inc. D.W. Bristol, Ph.D. G.P. Flake, M.D. J.R. Bucher, Ph.D. National Toxicology Program L.T. Burka, Ph.D. R.A. Herbert, D.V.M., Ph.D. R.S. Chhabra, Ph.D. National Toxicology Program R.A. Herbert, D.V.M., Ph.D. G.D. Hill, D.V.M., Ph.D. A.P. King-Herbert, D.V.M. National Toxicology Program G.E. Kissling, Ph.D. K. Kuettler, D.V.M., F.T.A., Observer D.E. Malarkey, D.V.M., Ph.D. BASF, Germany D.P. Orzech, M.S. D.E. Malarkey, D.V.M., Ph.D. S.D. Peddada, Ph.D. National Toxicology Program C.S. Smith, Ph.D. G. Pearse, B.V.M.&S. National Toxicology Program G.S. Travlos, D.V.M. C.C. Shackelford, D.V.M., M.S., Ph.D. K.L. Witt, M.S. Experimental Pathology Laboratories, Inc. J.C. Peckham, D.V.M., M.S., Ph.D. Battelle Columbus Operations Experimental Pathology Laboratories, Inc. Conducted studies and evaluated pathology findings A. Veit, Observer Experimental Pathology Laboratories, Inc. M.R. Hejtmancik, Ph.D., Principal Investigator M.J. Ryan, D.V.M., Ph.D. Evaluated slides and prepared pathology report on mice (May 31, 2005) D.M. Sells, D.V.M., Ph.D. M.A. Hanes, D.V.M., Chairperson Experimental Pathology Laboratories, Inc. ILS, Inc. Provided pathology review M.F. Cesta, D.V.M. National Toxicology Program M.H. Hamlin, II, D.V.M., Principal Investigator K.J. Cimon, D.V.M., M.S. K.J. Cimon, D.V.M., M.S. Experimental Pathology Laboratories, Inc. J.C. Peckham, D.V.M., M.S., Ph.D. G.P. Flake, M.D. 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 G.D. Hill, D.V.M., Ph.D. Prepared quality assessment audits National Toxicology Program K. Kuettler, D.V.M., F.T.A., Observer S. Brecher, Ph.D., Principal Investigator BASF, Germany D.E. Malarkey, D.V.M., Ph.D. National Toxicology Program G. Pearse, B.V.M.&S. National Toxicology Program J.C. Peckham, D.V.M., M.S., Ph.D. Experimental Pathology Laboratories, Inc. Formamide, NTP TR 541 3 Constella Group, Inc. Biotechnical Services, Inc. Provided statistical analyses Prepared Technical Report D.W. Crockett, Ph.D., Principal Investigator S.R. Gunnels, M.A., Principal Investigator L.J. Betz, M.S. B.F. Hall, M.S. K.P. McGowan, M.B.A. L.M. Harper, B.S. J.I. Powers, M.A.P. D.C. Serbus, Ph.D. 4 CONTENTS ABSTRACT . 7 EXPLANATION OF LEVELS OF EVIDENCE OF CARCINOGENIC ACTIVITY................. 10 TECHNICAL REPORTS REVIEW SUBCOMMITTEE........................................ 11 SUMMARY OF TECHNICAL REPORTS REVIEW SUBCOMMITTEE COMMENTS............. 12 INTRODUCTION . 13 MATERIALS AND METHODS . 17 RESULTS ............................................................................... 29 DISCUSSION AND CONCLUSIONS........................................................ 57 REFERENCES........................................................................... 61 APPENDIX A Summary of Lesions in Male Rats in the 2-Year Gavage Study of Formamide . 65 APPENDIX B Summary of Lesions in Female Rats in the 2-Year Gavage Study of Formamide . 79 APPENDIX C Summary of Lesions in Male Mice in the 2-Year Gavage Study of Formamide . 91 APPENDIX D Summary of Lesions in Female Mice in the 2-Year Gavage Study of Formamide . 107 APPENDIX E Genetic Toxicology . 121 APPENDIX F Clinical Pathology Results . 131 APPENDIX G Organ Weights and Organ-Weight-to-Body-Weight Ratios . 139 APPENDIX H Reproductive Tissue Evaluations and Estrous Cycle Characterization . 145 APPENDIX I Chemical Characterization and Dose Formulation Studies ........................ 149 APPENDIX J Ingredients, Nutrient Composition, and Contaminant Levels in NTP-2000 Rat and Mouse Ration........................................... 163 APPENDIX K Sentinel Animal Program.................................................... 167 Formamide, NTP TR 541 5 SUMMARY Background Formamide has a variety of uses, including as a softener for paper, gums, and glues. We studied the effects of formamide on male and female rats and mice to identify potential toxic or cancer-related hazards. Methods We deposited solutions containing formamide in deionized water directly into the stomachs of male and female rats and mice. Groups of 50 animals received 20, 40, or 80 milligrams of formamide per kilogram body weight five days per week for two years; groups of animals receiving water alone served as the control groups. At the end of the study, tissues from more than 40 sites were examined for every animal. Results The only effects observed in male and female rats were lower body weights in animals receiving the highest dose of formamide and a slight increase in bone marrow hyperplasia in male rats. There was an increased occurrence of liver hemangiosarcomas in all three groups of male mice receiving formamide and a less pronounced increase in hepatocellular adenomas or carcinomas of the liver in female mice. In male mice receiving 80 mg/kg of formamide, there was mineralization in the testes. Conclusions We conclude that exposure to formamide did not cause cancer in male or female rats. Exposure to formamide did produce hemangiosarcomas in the liver of male mice and may have been related to an increase in liver tumors in female mice. Increased incidences of bone marrow hyperplasia in male rats and testicular mineralization in male mice were also associated with formamide exposure. 6 Formamide, NTP TR 541 7 ABSTRACT FORMAMIDE CAS No. 75-12-7 Chemical Formula: CH3NO Molecular Weight: 45.04 Synonyms: Carbamaldehyde; formic acid, amide; formimidic acid; methanamide Formamide is used as a softener for paper, gums, and for 14 weeks. Additional groups of 10 male and 10 animal glues; as an ionizing solvent; and in the manufac- female rats (clinical pathology study) and five male ture of formic esters and hydrocyanic acid. Formamide and five female rats (plasma concentration study) were was nominated for reproductive and genetic toxicity administered the same doses, 5 days per week for up to evaluation by the Environmental Defense Fund and 14 weeks. All core study rats survived to the end of the for carcinogenicity evaluation by the National Cancer study. Mean body weights of females in the 40 mg/kg Institute because of the potential for human expo- group and males and females in the 80 and 160 mg/kg sure associated with its widespread industrial use, the groups were significantly less than those of the vehicle absence of data adequately characterizing its potential controls.
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