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TR-565: Milk Thistle Extract (CASRN 84604-20-6) in F344/N Rats And

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TR-565: Milk Thistle Extract (CASRN 84604-20-6) in F344/N Rats And NTP TECHNICAL REPORT ON THE TOXICOLOGY AND CARCINOGENESIS STUDIES OF MILK THISTLE EXTRACT (CAS NO. 84604-20-6) IN F344/N RATS AND B6C3F1 MICE (FEED STUDIES) NATIONAL TOXICOLOGY PROGRAM P.O. Box 12233 Research Triangle Park, NC 27709 May 2011 NTP TR 565 NIH Publication No. 11-5907 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 MILK THISTLE EXTRACT (CAS NO. 84604-20-6) IN F344/N RATS AND B6C3F1 MICE (FEED STUDIES) NATIONAL TOXICOLOGY PROGRAM P.O. Box 12233 Research Triangle Park, NC 27709 May 2011 NTP TR 565 NIH Publication No. 11-5907 National Institutes of Health Public Health Service U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES 2 CONTRIBUTORS National Toxicology Program Dynamac Corporation Evaluated and interpreted results and reported findings Prepared quality assessment audits J.K. Dunnick, Ph.D., Study Scientist S. Brecher, Ph.D., Principal Investigator A. Nyska, D.V.M., Study Pathologist S. Iyer, B.S. J.B. Bishop, Ph.D. V.S. Tharakan, D.V.M. J.R. Bucher, Ph.D. R.S. Chhabra, Ph.D. NTP Pathology Working Group P.M. Foster, Ph.D. Evaluated slides and contributed to pathology report R.A. Herbert, D.V.M., Ph.D. on 2-year rats (January 22 and 31, 2008) M.J. Hooth, Ph.D. A.P. King-Herbert, D.V.M. S. Newbigging, M.Sc., D.V.M., D.V.Sc., Coordinator G.E. Kissling, Ph.D. ILS, Inc. D.E. Malarkey, D.V.M., Ph.D. N. Allison, D.V.M. Experimental Pathology Laboratories, Inc. J.H. Roycroft, Ph.D. D. Dixon, D.V.M., Ph.D. J.M. Sanders, Ph.D. National Toxicology Program B.P. Singh, D.V.M. S.A. Elmore, D.V.M., M.S. C.S. Smith, Ph.D. National Toxicology Program G.S. Travlos, D.V.M. G.P. Flake, M.D. N.J. Walker, Ph.D. National Toxicology Program K.L. Witt, M.S. R.A. Herbert, D.V.M., Ph.D. National Toxicology Program D.E. Malarkey, D.V.M., Ph.D. Southern Research Institute National Toxicology Program Conducted studies and evaluated pathology findings A. Nyska, D.V.M. ILS, Inc. C.D. Hébert, Ph.D., Principal Investigator J.C. Peckham, D.V.M., M.S., Ph.D. J.E. Heath, D.V.M. Experimental Pathology Laboratories, Inc. J.F. Mann, B.S., D.V.M. B.P. Singh, B.V.Sc., M.S. National Toxicology Program Experimental Pathology Laboratories, Inc. Provided pathology review M.H. Hamlin, II, D.V.M., Principal Investigator N. Allison, D.V.M. H.M. Kolenda-Roberts, D.V.M., Ph.D. J.C. Peckham, D.V.M., M.S., Ph.D. TherImmune Research Corporation Provided SMVCE analysis G.W. Wolfe, Ph.D., Principal Investigator H.S. Seung, M.S. Milk Thistle Extract, NTP TR 565 3 NTP Pathology Working Group SRA International, Inc. Evaluated slides and contributed to pathology report Provided statistical analyses on 2-year mice (December 6 and 13, 2007) P.W. Crockett, Ph.D., Principal Investigator P.E. Blackshear, D.V.M., Ph.D., Coordinator L.J. Betz, M.S. ILS, Inc. K.P. McGowan, M.B.A. D. Dixon, D.V.M., Ph.D. National Toxicology Program S.A. Elmore, D.V.M., M.S. Biotechnical Services, Inc. Prepared Technical Report National Toxicology Program G.P. Flake, M.D. S.R. Gunnels, M.A., Principal Investigator National Toxicology Program R.A. Herbert, D.V.M., Ph.D. B.F. Hall, M.S. National Toxicology Program L.M. Harper, B.S. K. Hobbie, D.V.M. J.I. Powers, M.A.P. ILS, Inc. D.C. Serbus, Ph.D. H.M. Kolenda-Roberts, D.V.M., Ph.D. Experimental Pathology Laboratories, Inc. D.E. Malarkey, D.V.M., Ph.D. National Toxicology Program A. Nyska, D.V.M. ILS, Inc. J.B. Nold, D.V.M. GlaxoSmithKline J.C. Peckham, D.V.M., M.S., Ph.D. Experimental Pathology Laboratories, Inc. B.P. Singh, B.V.Sc., M.S. National Toxicology Program 4 CONTENTS ABSTRACT ................................................................................................................................................................. 7 EXPLANATION OF LEVELS OF EVIDENCE OF CARCINOGENIC ACTIVITY ........................................ 11 TECHNICAL REPORTS REVIEW SUBCOMMITTEE ..................................................................................... 12 SUMMARY OF TECHNICAL REPORTS REVIEW SUBCOMMITTEE COMMENTS ................................ 13 INTRODUCTION ..................................................................................................................................................... 15 MATERIALS AND METHODS .............................................................................................................................. 25 RESULTS ................................................................................................................................................................... 35 DISCUSSION AND CONCLUSIONS ..................................................................................................................... 59 REFERENCES .......................................................................................................................................................... 61 APPENDIX A Summary of Lesions in Male Rats in the 2-Year Feed Study of Milk Thistle Extract ................................................................................................................. 71 APPENDIX B Summary of Lesions in Female Rats in the 2-Year Feed Study of Milk Thistle Extract ................................................................................................................. 85 APPENDIX C Summary of Lesions in Male Mice in the 2-Year Feed Study of Milk Thistle Extract ................................................................................................................ 99 APPENDIX D Summary of Lesions in Female Mice in the 2-Year Feed Study of Milk Thistle Extract .............................................................................................................. 111 APPENDIX E Genetic Toxicology ...................................................................................................................... 123 APPENDIX F Clinical Pathology Results .......................................................................................................... 135 APPENDIX G Organ Weights and Organ-Weight-to-Body-Weight Ratios ................................................... 143 APPENDIX H Reproductive Tissue Evaluations and Estrous Cycle Characterization ................................. 147 APPENDIX I Chemical Characterization and Dose Formulation Studies ................................................... 151 APPENDIX J Feed and Compound Consumption in the 2-Year Feed Studies of Milk Thistle Extract ............................................................................................................... 165 Milk Thistle Extract, NTP TR 565 5 APPENDIX K Ingredients, Nutrient Composition, and Contaminant Levels in NTP-2000 Rat and Mouse Ration .......................................................................................... 171 APPENDIX L Sentinel Animal Program ..........................................................................................................
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