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NTP Technical Report on Toxicity Studies of Dibutyl Phthalate

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NTP Technical Report on Toxicity Studies of Dibutyl Phthalate National Toxicology Program Toxicity Report Series Number 30 NTP Technical Report on Toxicity Studies of Dibutyl Phthalate (CAS No. 84-74-2) Administered in Feed to F344/N Rats and B6C3F1 Mice Daniel S. Marsman, D.V.M., Ph.D., Study Scientist National Toxicology Program Post Office Box 12233 Research Triangle Park, NC 27709 NIH Publication 95-3353 March 1995 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. 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. 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 919/541-1371). NTP Central Data Management NIEHS Post Office Box 12233 Research Triangle Park, NC 27709 National Toxicology Program Toxicity Report Series Number 30 NTP Technical Report on Toxicity Studies of Dibutyl Phthalate (CAS No. 84-74-2) Administered in Feed to F344/N Rats and B6C3F1 Mice Daniel S. Marsman, D.V.M., Ph.D., Study Scientist National Toxicology Program Post Office Box 12233 Research Triangle Park, NC 27709 NIH Publication 95-3353 March 1995 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. United States Department of Health and Human Services Public Health Service National Institutes of Health 2 DIBUTYL PHTHALATE, NTP TOXICITY REPORT NUMBER 30 CONTRIBUTORS This NTP report on the toxicity studies of dibutyl phthalate is based primarily on studies that took place from October 1987 through November 1989. National Toxicology Program Experimental Pathology Laboratories, Inc. Evaluated experiment, interpreted results, and reported findings Provided pathology quality assessment Daniel S. Marsman, D.V.M., Ph.D., John Peckham, D.V.M., M.S., Ph.D. Study Scientist Gary Riley, M.V.Sc., Ph.D. John R. Bucher, Ph.D. Robert E. Chapin, Ph.D. Michael R. Elwell, D.V.M., Ph.D. Environmental Health Research and Testing, Inc. Joel Mahler, D.V.M. Provided sperm morphology, vaginal cytology evaluation, and reproductive H. B. Matthews, Ph.D. assessment by continuous breeding (rats) Joseph H. Roycroft, Ph.D. Leta Hommel Barnes Michael D. Shelby, Ph.D. Teresa Cocanougher, B.A. Gregory S. Travlos, D.V.M. Dushyant K. Gulati, Ph.D. Kristine L. Witt, M.S. Susan Russell, B.A. Oak Ridge Associated Universities Research Triangle Institute Battelle Columbus Laboratories Provided reproductive assessment by continuous breeding (mice) Principal contributors James C. Lamb IV, Ph.D. Perry J. Kurtz, Ph.D., Principal Investigator A. Davis Lawton, M.S. Arthur C. Peters, D.V.M., Jerry R. Reel, Ph.D. Principal Investigator Joanne M. Killinger, Ph.D. Lawrence E. Mezza, D.V.M. Analytical Sciences, Inc. Ronald L. Persing, D.V.M. Provided statistical analyses Michael J. Ryan, D.V.M., Ph.D. John D. Toft II, D.V.M., M.S. Sarah R. Kennedy, M.S. Steven Seilkop, M.S. Janet L. Teague, M.S. NTP Pathology Working Group Evaluated slides and prepared pathology report Biotechnical Services, Inc. Chair Paul K. Hildebrandt, D.V.M., Provided toxicity report preparation PATHCO, Inc. Michael R. Elwell, D.V.M., Ph.D. C. Michael Bailey, B.S. Pharm., National Toxicology Program Principal Investigator Jeffrey Everitt, D.V.M. Janet L. Elledge, B.A. Chemical Industry Institute of Toxicology Daphne D. Lambright, Ph.D. William F. MacKenzie, D.V.M., M.S. Waynette D. Sharp, B.A., B.S. Experimental Pathology Laboratories, Inc. Joel Mahler, D.V.M. National Toxicology Program Cynthia C. Shackelford, D.V.M., M.S., Ph.D. National Toxicology Program DIBUTYL PHTHALATE, NTP TOXICITY REPORT NUMBER 30 3 PEER REVIEW The draft report on the toxicity studies of dibutyl phthalate was evaluated in July 1994 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. Janardan K. Reddy, M.D. Mary Jo Vodicnik, Ph.D. Department of Pathology Lilly MSG Development Center Northwestern University Medical School Belgium Chicago, IL 4 DIBUTYL PHTHALATE, NTP TOXICITY REPORT NUMBER 30 TABLE OF CONTENTS ABSTRACT ................................................................................. 5 INTRODUCTION ............................................................................ 13 Physical and Chemical Properties, Production, Use, and Exposure ............................... 13 Metabolism and Disposition .............................................................. 14 Toxicity ............................................................................... 15 Study Rationale and Design ............................................................... 19 MATERIALS AND METHODS .................................................................. 23 Procurement and Characterization of Dibutyl Phthalate ........................................ 23 Dose Formulations ...................................................................... 23 Toxicity Study Designs ................................................................... 24 Genetic Toxicity Studies .................................................................. 34 Statistical Methods ...................................................................... 36 Quality Assurance ....................................................................... 37 RESULTS .................................................................................. 39 Maximum Perinatal Exposure Determination Feed Study in F344/N Rats ........................ 39 13-Week Feed Study with Perinatal Exposure in F344/N Rats .................................. 45 13-Week Feed Study in F344/N Rats ....................................................... 62 Continuous Breeding Study in Sprague-Dawley Rats ........................................... 71 Perinatal Peroxisome Determination Studies in F344/N Rats ................................... 73 Maximum Perinatal Exposure Determination Feed Study in B6C3F1 Mice ......................... 75 13-Week Feed Study in B6C3F1 Mice ....................................................... 81 Continuous Breeding Study in Swiss (CD-1® ) Mice ............................................ 86 Genetic Toxicity ........................................................................ 86 DISCUSSION ............................................................................... 87 REFERENCES .............................................................................. 93 APPENDIXES Appendix A Summary of Nonneoplastic Lesions in Rats and Mice .......................... A-1 Appendix B Organ Weights and Organ-Weight-to-Body-Weight Ratios ...................... B-1 Appendix C Hematology and Clinical Chemistry Results .................................. C-1 Appendix D Reproductive Tissue Evaluations and Estrous Cycle Characterization ........................................ D-1 Appendix E Continuous Breeding Studies .............................................. E-1 Appendix F Perinatal Peroxisome Determination Studies ................................. F-1 Appendix G Genetic Toxicology ...................................................... G-1 DIBUTYL PHTHALATE, NTP TOXICITY REPORT NUMBER 30 5 ABSTRACT
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