NATIONAL TOXICOLOGY PROGRAM Technical Report Series No. 427 TOXICOLOGY AND CARCINOGENESIS STUDIES OF TURMERIC OLEORESIN (CAS NO. 8024-37-1) (MAJOR COMPONENT 79%-85% CURCUMIN, CAS NO. 458-37-7) IN F344/N RATS AND B6C3Ft MICE (FEED STUDIES) U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES Public Health Service 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 Human Services (DHHS): the National Cancer Institute (NCI), National Institutes of Health; *theNational Institute of Environmental Health Sciences (NIEHS), National Institutes of Health; the National Center for Toxicological Research (NCTR),Food and Drug Administration; and the National Institute for Occupational Safety and Health (NIOSH), Centers for 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 relatingto 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 thisTechnical Report were performed underthe direction of the NIEHS and were conducted in compliance withNTP laboratory health and safety requirements and must meet or exceed all applicable federal, state, and local health and safety regulations. Animal care and use were in accordance with the Public Health Service Policy on Humane Care and Use of Animals. The prechronic and chronic studies were conductedin compliance with Food and Drug Administration (FDA) Good Laboratory Practice Regulations, andaall aspectsof the chronic studies were subjected to retrospective quality assurance audits before being presented for publicreview. These studies are designed and conducted to characterize and evaluate the toxicologic potential, including carcinogenic activity, of selected chemicals in laboratory animals (usually two species, rats and mice). Chemicals selected for NTP toxicology and carcinogenesis studies are chosen primarily on thebases of human exposure, level of production, and chemical structure. Selection per se is not an indicator of a chemical's carcinogenic potential. These NTP Technical Reports are available for salefrom the National Technical InformationService, 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). ~~ - ~ 2 Turmeric Oleoresin, NTP TR 427 CONTRIBUTORS National Toxicology Program NTP Pathology Working Group Evaluated sliaks, prepared pathology report on rats Evaluated and interpreted results and reported @din@ (30April 1991) C.J. Alden, Ph.D. R.M. Lovatch, D.V.M., Chair G.A. Boorman, D.V.M., Ph.D. Pathology Associates, Incorpo,rated D.A. Bridge, B.S. M.P. Jokinen, D.V.M. J.K. Dunnick, Ph.D. National Toxicology Program S.L. Eustis, D.V.M., Ph.D. C.M. Keenan, V.M.D. T.J. Goehl, Ph.D. Merck, Sharp & Dohme R.A. Griesemer, D.V.M., Ph.D. M.M. McDonald, D.V.M., Ph.D. J.K. Haseman, Ph.D. National Toxicology Program C.C. M.S., 'R.D. Irwin, Ph.D. Shackelford, D.V.M., Ph.D. G.N. National Toxicology Program Rao, D.V.M., Ph.D. P.G. Stromberg, D.V.M., Ph.D. R.C. Sills, D.V.M., Ph.D. Ohio State University D.B. Walters, Ph.D. K. Yoshitomi, D.V.M., Ph.D. KL.Witt, M.S., Oak Ridge Associated Universities Experimental Pathology Laboratories, Inc. EG&G Mason Research Institute Evaluated slides, prepared pathology report on mice Conducted studies, evaluated pathology findngs (28 March 1991) H.S. Lilja, Ph.D., PrincipalInvestigator J.C. Seely, D.V.M., Chair R.W. Fleischman, D.V.M. PATHCO, Incorporated G.A. Boorman, D.V.M., Ph.D. L.E. Sendelbach, Ph.D. National Toxicology Program R.L. Taber, Ph.D. J.F. Hardisty, D.V.M. National Toxicology Program Experimental Pathology Laboratories, Inc. M.P. Jokinen, D.V.M. Provided parhology quality assurance National Toxicology Program R.S. Miller, D.V.M. J.F. Hardisty, D.V.M., PrincipalInvestigator Chemical Industry Institute of Toxicology K. Yoshitomi, D.V.M., Ph.D. T.S. Peters, D.V.M. (observer) Food and Drug Administration Integrated Laboratory Systems C.C. Shackelford, D.V.M., M.S., Ph.D. Prepared quality aswance audits National Toxicology Program S.L. Smith, J.D., PrincipalInvestigator Biotechnical Services, Inc. Prepared Technical Report D.D. Lambright, Ph.D., PrincipalInvestigator P. Chaffin, M.S. G.F. Corley, D.V.M. A.B. James-Stewart, B.S. 3 ABSTRACT ................................................................... 5 EXPILANATION OF LEVELS OF EVIDENCE OF CARCINOGENIC ACTIVITY . .. .... .. no TECHBNIC~REPORTS wmmSUBCOMMITTEE . .. ... nn SUMWY OF TECHBNHCAILREPORTS REVIEW SUBCOMMI~EECOMMENTS . ... n2 INTRODUCTION . .. ni MATERIALS AND METHODS .. .. 17 RESULTS .................................................................... 25 DHSCUSSHON AND CONCLUSIONS . .. .. .. :. 47 REFERENCES .... .. ... ... .. ..... ! . 51 APPENDIXA Summary off Lesions in Male Rats in the 2-Year Feed Study OP Turmeric Oleoresin ...... .. .. ..... 57 APPENDIXB Summary of Lesions in FemaleRats in the 2-Year Feed Study of Turmeric Oleoresin . .. .. .. 107 APPENDIXC Summary off Lesions in Male Mice in the 2-Year Feed Study off ~nameric01eoresin ... .. .. .. .. 107 APPENDIXD Summary OP Lesions in Female Mice in the 2-Year Feed Study of ~urmeric~~eoresiaa. .. ... nsJ APPENDIXE Genetic Toxicology . .. ... .. 223 APPENDIXF Organ Weightsand Organ- eight-to-Bod~WeighP Ratios . ... 231 APPENDIXG Hematology, Clinical Chemistry,andUrinalysisResults . ... ... 237 APPENDIXH ChemicaU Characterization and Dose Formulations . .. .. 2511 APPENDIXII Feed and Compound Consumption . .. 263 APPENDIXK Sentinel Animal Program . .. 275 TURMlERIC OlLIEORlESlIN CAS NO.8024-37-1 Synonyms: curcuma oil; oil of turmeric; turmeric oil; curcuma longa oils; curcuma long oil; Curcumin (CURCUMIN CAS NO.458-37-7 Chemical Formula: q,H,O, MolecularWeight: 368.37 Synonyms: 1,7-Bis(4-hydroxy-3-methoxyphenyl)-l,6-heptadiene-3,5-dione;C.I. Natural Yellow 3; C.I.7530; Curcuma; diferuloylmethane; E 100; Haidr; Halad; Haldar, Halud; HSDB 4334;Indian Saffron: kacha haldi; Kurkumin; menta earth; Souchet; Turmeric Yellow; yellow ginger, yellow root; Yo-kin; Zlut Prirodni 3; NCLC613253 Turmeric oleoresin is the organic extractof turmeric, thanthat of the controls.Feedconsumption by a ground powder fromthe root of the Curcuma plant, exposed male and female rats was similar to that by and is added to food items as a spice and coloring the controls. Dietary levels of 1,000, 5,UOO, 10,000, agent.Turmericoleoresin,turmeric,andcurcumin 25,000, or 50,UOO ppm turmeric oleoresin were esti- (the major componentfound in turmeric) were nomi- mated to deliver average daily doses of 50, 250, 480, nated by the National Cancer Institute and the Food 1,300, or 2,600 mghg body weight to males and 60, andDrugAdministrationfor study becausethese 300, 550, 1,450, or 2,80 mghgto females. The chemicals are used in food items and curry powders, absolute andrelative liver weights of female rats and andthere was littleinformation on their toxic or the relative liver weights of male rats receiving 5,OOO, carcinogenicproperties. Pure curcumin was not lO,UOO, 25,UOO, and 50,OOO ppmwere significantly availablein sufficient quantitiesfor study, anda greaterthanthose of the controls. There were no turmericoleoresinwitha high curcumincontent biologically significant differencesinhematologic, (79% to 85%) was selected for evaluation. Toxicity clinical chemistry, or urinalysis parameters. Clinical and carcinogenicity studies were conductedby admin- findings included stained fur, and discoloredfeces and istering turmeric oleoresin in feed to groups of male urine of exposed animals, presumably due to the andfemale F344/BJ ratsand B6C3F, mice for parent compound or its metabolites. Hyperplasia of 13 weeks and 2 years. Genetic toxicology studies the mucosal epithelium was observed in the cecum wereconducted in Salmonella typhimurium and andcolon of maleandfemalerats that received cultured Chinese hamster ovary cells. 50,UOO ppm. n w m STUDY~ IN RATS Groups of 10 male and 10 female F344/Ed rats were nwmKSTUDY IN MICE fed diets containing 0, 1,000,5,000, 10,000, 25,UOO, or Groups of 10 male and 10 female B6C3F, mice were 50,OOO ppmturmericoleoresin.Allrats survived fed diets containing 0, l,OOO, 5,0,10,000, 25,000, or until the end of the study. The final mean body 50,000 ppm turmericoleoresin. There were no weight of males receiving 50,000 ppm was 5% lower deaths attributed to turmeric oleoresin and the final 6 Turmeric Oleoresin, NTP TR 427 mean body weight gains and final mean body weights Hematology and Clinical Chemistry of all exposed groups of male and female mice were In male andfemalerats receiving 50,OOO ppm the similar to those of the controls. Feed consumption hematocrit values, hemoglobin concentrations,and by exposed male and female mice was similar to that erythrocytecounts atthe 15-monthinterim evalu- by the controls. Dietary levels of l,OOO, 5,OOO, lO,OOO, ation were significantly lower than those in thecon- 25,OOO, or 50,OOO ppm turmeric oleoresin were esti- trols. In addition, platelet counts in