Document ZG58n6wQNQKbjYd2BXVrGo80

AR226-3018 y Study Title Repeated Dose Oral Toxicity: 14-Day Feeding Study with H-19683 in Male Mice Author John V. Sarver Study Completed On April 28, 1995 Performing Laboratory Haskell E. I. du Font de Nemours and Company Laboratory for Toxicology and Industrial Elkton Road, P.O. Box 50 Newark, Delaware 19714 Medicine Laboratory Project ID Haskell Laboratory Report Mo. 325-95 Page 1 of 23 Company Sanitized, Does not contain TSCA CBI DuPont SIS. 325-95 REPEATED DOSE ORAL TOKtCITY; 14-DAY FEEDING STUDY tfITH H-19683 IN HALE MICE GENERAL INFORMATION Substance Tested: Synonyms/Codes; Purity; Physical Form: Composition; Contaminants; C.A.S. Registry No.; Sponsor; Study Initiated/Completed; In-life Study Initiated/Completed; DuPont Chemicals/DuPont Polymers E. I. du Font de Nemours and Company Vilmington, Delaware 10/8/92 - 4/28/95 11/4/92 - 11/18/92 2 - Company Sanitized. Does not contain TSCA CBI DuPont HLR 325-95 REPEATED DOSE ORAL TOXICITY: 14-DAY FEEDING STUDY VITH H-19683 IN MALE MICE The purpose of this study was to evaluate the repeated dose oral toxicity of H-196P3 when incorporated into nutritionally adequate chow and fed to sale Crl;CD-l*BR mice for 14 consecutive days. Mice vere fed diets containing 0, 3, 30, 300, or 3000 ppa H-19683. Mice were weighed and observed for clinical signs of toxicity once each week. Food consunption was not measured. At the end of the 2 week feeding period, all nice were euthanatized and necropsied. The livers were weighed, and liver weight/final body weight ratios were calculated. No deaths occurred during the study. Other than a body weight effect ii mice from the 3000 ppm dose group, no adverse clinical signs of toxicity ve-e observed during the study. Significant decreases in mean body weight were observed in mice from the 3000 ppm dose group on test days 7 and 14. Statistically significant decreases in mean body weight gain were observed in this same dose group for the intervals 0-7 days on test, and overall. Significant increases in mean absolute and relative liver weights were observed in mice from the 300 and 3000 ppm dose groups killed at the end! of the 14-day feeding period. The no observable adverse effect level (NOAEL) was determined to te 30 ppm. The NOAEL was based on the increases in mean absolute and relative liver weight observed in the 300 and 3000 ppm dose groups and the decreases in ..ean body weights and body weight gains observed in the 3000 ppin dose group. Work by: ^Y^Ui If --Ai^-lf^ 4 f^-WS' Tracy A. Pillib^n Toxicology Technician Study Director: ^&^-^ L^ . ^CU^e^ --------John V. Sarver^ Toxicologist ^ '3^ ^ JVS/alw 3 -- -- Company Sanitized. Does not contain TSCA CB DuPont HLR 325-95 RgPEATED DOSE ORAL TOXICITY: 14-DAY FEEDING STUDY WITH H-196B3 IS MALE MICE jsrssaafKsiw The purpose of this study was to evaluate the repeated dose oral toxicity of H-19683 when incorporated into nutritionally adequate chow and fed to male Crl:CDBR mi:e for 14 consecutive days. Mice were fed diets contained 0, 3, 30, 300, or 3000 ppffl H-19683. MATERIALS AND HETBODS A. Test Substance The test substance, H-19683, vas supplied by DuPont Specialty Cheoicals, E. es t I. im a du ted F on to t de be Nem M^ ours lf-- and -- Coflp --^ a ny, Av Vi ail lm ab i l n e g t i o n n, fo r Del mat awa ion re. ab o P ut urity the i was den t i t y , purity, compos i tTon-stabT^Ryand toxicity of the test substance was filed with Haskell Laboratory Information Services by the Sponsor when the substance was submitted for testing. Upon receipt, the test substance was assigned a unique H-sskell Number (H-19683). B. Test Species On 10/27/92, 30 male Crl:CD-l(ICR)BR mice, born approximately 9/21/92, were received from Charles River Laboratories, Inc., Montreal, Canada. On 11/4/92, the body weights ranged from 24.4 to 29.6 grams. The Crl;CD-lBR mouse was selected on the basis of extensive experience with this strain and its suitability with respect to hardiness, longevity, sensitivity, and low incidence of spontaneous diseases. C. Animal Husbandry All mice were housed singly in suspended stainless steel, wire-mesh cages throughout the study. Animal rooms were maintained on a 12-hour light/dark cycle and targeted at a temperature of 23 + 2C and a relative humidity of 50 10X. Water was provided ad libitua from an automatic watering system. During the quarantine period (see Section 0), all mice were fed (ground) Purina^ Certified Rodent Chow9 15002. During the test period, all mice were fed the diet of their respective treatment group (see Section F). 4 - - Company Sanitized. Does not contain TSCA CBI DuPont BLR 325-95 Haskell Laboratory has an animal health monitoring program. This program is monitored and administered analyzed for by the Laboratory Veterinarian. Water samples total bacterial counts and for the presence of are periodically and other contaminants. Additionally, samples from freshly coliforns, lead, washed cages and cage racks are periodically analyzed to assure adequate sanitation by the cagewashers. Data from this program are maintained Animal feed is certified by the manufacturer separately from study records. to oeet specified nutritional requirements and to be free of a list of specified contaminants. On the basis of these analyses, there is no evidence were present in the feed or water in amounts suggesting that contaminants which may have interfered with the results of this study. D. Quarantine Period Upon arrival at Haskell Laboratory, the mice vere removed from shipping cartons and housed 3 per cage in suspended stainless steel, wire-mesh mouse was identified by color-code cages. tail During the quarantine period, each and a temporary identification number affixed to its cage. The marKing ani-aals were weighed 3 times during the quarantine period and observed daily for any gross signs of disease or injury. On the basis of body weights and clinical observations, all mice were released from quarantine on 11/3/92 by the laboratory veterinarian. E. Assignment to Groups and Study Start After the quarantine period, 25 mice were selected for study freedom from any clinical use on the signs of basis of adequate body weight gain and disease or injury. The selected mice were divided by computerized, stratified randomization into S groups of 5 significant differences among mice each, group body so there were weight means. no The statistically of mice within each group was randomized. Each group of mice was order designated as either a control, low-, low-intermediate-, or high-concentration group. After assignment to dosing groups, each mouse was assigned a 5-digit number was printed on a card affixed to the Houa:sskiedlel aonf imthael ncuammgaerbk.eerd. EoaEnchacthhmeoumseouwsaes'saltsaoilawssiitghnewdataer 3i-ndslgoliut bilde einntkif. icaTthieon number that was mice were approximately 44 days of age at study start. Mice not assigned to a test group were sacrificed by CO; asphyxiation. F. Concentration Selection and Diet Administration H-19683 was 3000 mixed in These the diet at concentrations of 0 concentrations were selected so (control), 3, 30, that the results of 300. this or study ppm. could be compared to the results obtained from an earlier study conducted with C-8. 5 - Company Sanitized. Does nol conla'inTSCA CBt DuPont BLR 325-95 During the test period, mice in each group were fed a diet of ground Purina* Certified Rodent Chow* 15002 that contained the following concentrations of H-19683: Group No. Male I III V VII IX Dietary Concentration* ___ppm___ t Rats/Group 0 (Control) 5 3 (Low) 5 30 (Lov-Intermediate) 5 300 (Intermediate) 5 3000 (High) 5 Weight/weight concentration of B-19683 G. Diet Preparation and Sampling For each diet concentration, thcehowapapnrdopmriixaeted abmy ohuanntdouf nHt-i1l96i8t3 awppaseafreirdst ahdodmeodgetnoeoauss. maTlhl eamhaonudn-tmoixfedgropuonrdtion was then added to the remainder of the ground chow weighed out for that diet in a Hobart low speed mixer and was mixed for 15 minutes. After mixing, the diet was visually inspected to ensure period it of was free of clumps. time. The diets for Control diets were also mixed for the same this study were prepared the day the study shotamrotegdenaenitdy, satonrdedstraebfirliigteyrawteerde utankteinl ufoser da. naSlyasmepslesat foa r lactoenrcednattreatiiofnd, eemed necessary by the study director. H. Body Weights and Clinical Observations All mice were weighed and observed for clinical signs each week throughout the study. Cageside examinations to dead mice were conducted twice daily. of toxicity once detect moribund or I. Food Consumption, Food Efficiency, and Intake of H-19683 Pood consumption was not determined during this study. J. Necropsy At the end of the 2-week feeding mice were euthanatized by chloroform livers were weighed at necropsy, and were calculated. period, all mice were necropsied. The anesthesia and exanguination. The liver weight/final body weight ratios Company Sanitized. Does not contain TSCA CBI ,, DuPont HLR 325-95 STATISTICAL AKALTSES Body weights, body weight gains, and organ weights were analyzed by one-way analysis of variance. When the statistic for differences aaong group means (F statistic) was significant, pairwise comparisons between and control groups were made with the Dunnett's test. Significance was judged at alpha = 0.05. a test test RECORDS AND SAMPLE KETEWTlOW All original records will be retained at Haskell Laboratory or at the Records Management Center, E. I. du Font de Nenours and Company, Wilnington, Delaware. RESULTS A. Diet Sample Analysis All diets prepared appeared to be thoroughly mixed. No clumps or other signs of non-uniform mixing were observed. Therefore, diet samples taken at the time of diet preparation were discarded without analysis. B' Body Weights and Body Weight Gains (Tables 1-2', Appendix I) Statistically significant decreases in mean body weight of mice from the 3000 ppm dose group were observed on test days 7 and 14. Statistically significant decreases in mean body weight gains of this same group were observed for the interval 0-7 days on test and overall. C. Clinical Observations One mouse in the 3000 ppm dose group exhibited pallor of the ears on test days 7 and 14. No other clinical signs of toxicity were observed. D* Necropsy (Tables 3,4; Appendix II) Gross observations at necropsy revealed liver discoloration in one mouse from the 300 ppi dose group and in 4 nice from the 3000 ppm dose group. Significant increases in mean absolute and relative liver weights were observed in mice fron the 300 and 3000 ppm dose groups. No other gross observations were noted. 7 - Company Sanitized. Does not contain TSCA CW fret wn Hal ie* fed dit eontaininc coacrnti-atloM of 0, 30, 300, U Siniic*nt body v(fet ctfectc vr or 3000 pp 19683 for days. obfrvd in lee roa the 3000 pr done (roup. Sicnificwt lncrftc ia MU th 300 ad livr aluolutc end relative The wights cr obfrvcd io ice froB observable advrcc ffct lvl ob--rvd dutiox 3000 ffu da-- croups. no this study was 30 ppR. Company Sanitized. Does not contain TSCA CBjl OuPont BLR 325-95 ----^ Company Sanitized. Does nal eonlatnTSCA CW I J ----- REPEATED ORAI. nOSF. TOXTCTTY: 14 DAY PF.F.DTNf; STUDY WTTH H T)6R1 TN MAI.F. MTCR 1-ABLE 1 MEAN BODY WEIGHTS OF MALE NICE MEAN BODY WEIGHTS (g) I I I GROUP: I CONCENTRATION (PPH); 0 3 DAYS ON TEST 0 26.7( 0.5) 26.7< 1.9) 7 29.9( 1.5) 30.6( 1.5) 14 31.7( 1.3) 31.9( 1.3) V 30 27.2( 1.1) 29.6( 1.2) 31.5( 1.3) VII 300 27.8( 1.6) 29.8( 1.2) 31.0( 1.3) IX 3000 27.8( 0.6) 26.3( 2.4)* 28.7( 1.7)* ""Standard deviation is reported in parentheses Statistical aethods: One-way AOV and Dunnett's tests were performed on weekly interval data. (See Statistical Analyses) * Statistically significant difference from control at alpha = 0.05. Company Sanitized. D REPEATED ORAL DOSE TOXICITY: 14-DAY FEEDING STUDY WITH H-19683 IN MALE MICE TABLE 2 MEAN BODY WEIGHT GAINS OF HALE MICE MEAN BODY WEIGHT GAINS (g) GROUP: I CONCENTRATION (PPM)s 0 DAYS ON TEST 0- 7 3.2( l.2) 7-14 1.8( 0.5) 0-14 5.0( 0.9) III 3 3.9( 1,5) 1.4( 0.6) 5.3( 1.6) V 30 2.3( 1.1) 1.9( 0.6) 4.2( 1.2) VII IX 300 3000 2.0( 1.1) 1.2( 0.4) 3.2( 0.8) -1.5( 2.6)* 2.4( 1.5) 0.9( 2.0)* ""Standard deviation is reported In parentheses Statistical methods: One-way AOV and Dunnett's tests were performed on both weekly and summary interval data. (See Statistical Analyses) * Statistically significant difference from control at alpha = 0.05. Company Sanitized. Do DuPont BLR 325-95 REPEATED DOSE ORAL TOXICITY: 14-DAY FEEDING STUDY WITH H-19683 IN MALE NICE TABLE 3 MEAN FINAL BODY AND ORGAN WEIGHTS FROM MALE MICE MEAN FINAL BODY WEIGHTS (grans) GROUP CONC. (PPM) MEAN FINAL BODY WEIGHTS I 0 III 3 V 30 VII 300 IX 3000 31.7 ( 31.9 ( 31.5 < 31.0 ( 28.7 ( 1.3) 1.3) 1.3) 1.3) 1.7)# MEAN ABSOLUTE ORGAN WEIGHTS (grams) GROUP CONC. (PPM) LIVER I 0 1.811 (0.112) III 3 V 30 1.907 (0.154) 2.241 (0.344) VII 300 2.871 (0.336)# IX 3000 4.365 (0.685)# MEAN RELATIVE ORGAN WEIGHTS (X of body weight) GROUP CONC. (PPM) LIVER I 0 5.7043 (.1324) III 3 V 30 5.9781 (.5546) 7.0975 (.7963) VII 300 9.2486 (.9889) IX 3000 15.1789 (1.920)# Standard Deviation in parentheses # - Significantly different (p<0.05) from control group by Dunnett's Test 12 Company Sanitized. Does not contain TSCA CBS DuPont BLR 325-95 REPEATED DOSE ORAL TOXICITYs 14-DAY FEEDING STUDY WITH B-19683 IN HALE MICE TABLE 3 (continued) MEAN FINAL BODY AMD ORGAN WEIGHTS FROH HALE MICE MEAN FINAL BODY WEIGHTS (grans) GROUP CONC. (PPH) MEAN FINAL BODY WEIGHTS I 0 III 3 V 30 VII 300 IX 3000 31.7 31.9 31.5 31.0 28.7 (0.000) (0.803) (0.768) (0.444) (0.002)# TEST - HOMOGENEITY TEST - TREND BARTLETT'S TEST 0.008 0.002 0.972 MEAN ABSOLUTE ORGAN WEIGHTS (grams) GROUP CONC. (PPH) LIVER I 0 1.811 (0.000) III 3 V 30 1.907 (0.697) 2.241 (0.092) VII 300 2.871 (0.000)# IX 3000 4.365 (0.000)# TEST - HOMOGENEITY TEST - TREND BARTLETT'S TEST 0.000 0.000 0.011 Values in parentheses - P value of Student T Test comparison of treatment mean to control mean. - Significantly different (p<0.05) from control group by Dunnett's Test Homogeneity - P value of F Test of whether group means are equal. Trend - P value of F test of whether there is a dose-related change in group means. Bartlett's Test - P value of Test of Homogeneity of Variance. 13 Company Sanitized. Does not contain TSCA Gffl DuPont HLR 325-95 REPEATED DOSE ORAL TOXICITTi 14-DAY FEEDING STUDY WITH H-19683 IN MALE MICE TABLE 3 (continued) MEAN FINAL BODY AND ORGAN WEIGHTS FROM MALE MICE MEAN FINAL BODY WEIGHTS (gracs) GROUP CONC. (PPM) MEAN FINAL BODY WEIGHTS I 0 III 3 V 30 VII 300 IX 3000 31.7 31.9 31.5 31.0 28.7 (0.000) (0.803) (0.768) (0.444) (0.002)# TEST - HOMOGENEITY TEST - TREND BABTLETT'S TEST 0.008 0.002 0.972 MEAN RELATIVE ORGAK WEIGHTS (% of body weight) GROUP COKC. (P?X) LIVER I 0 5.7043 (0.000) III 3 V 30 5.9781 (0.688) 7.0975 (0.051) VII 30C 9.2486 (0.000)# IX 30C'? 15.1789 (0.000)# TEST - HOMOGENEITY TEST - TREND BARTLETT'S TEST 0.000 0.000 0.001 Values in parentheses - P value of Student T Test comparison of treatment mean to control mean. # - Significantly different (p<0.055 from control group by Dunnett's Test Homogeneity - P value of F Test of whether group means are equal. Trend - P value of F test of whether there is a dose-related change in group means. Bartlett's Test - P va3ue of Test of Hoaogeneity of Variance. - 14 - Company Sanitized. Does r.ol contain TSCACTO REFLATED ORAL DOSE TOXICITY: 14-DAY FEEDING STUDY WITH H-19683 IN HALE MICE TABLE 4 INCIDENCES OF GROSS OBSERVATIONS IN MALE MICE SITE/LESION; LIVER LARGE 5 5 DISCOLORATION GROUP DESIGNATION; 1 CONCENTRATION (PPM): 0 NUMBER IN GROUP: 5 III V VII 3 30 300 5 5 NOTE: 0 0 01 14 0 INCIDENCES REFLECT THE NUMBER OF ANIMALS WITH A GIVEN LESION. DuPont BIB 325-95 APPENDIX I INDIVIDUAL BODY WEIGHTS - 16 - Company Sanitized. Does nol contain TSCA CBI DuPont HLR 325-95 REPEATED DOSE ORAL TOXICITY: 14-DAY FEEDING STUDY WITH H-19683 IN HALE MICE APPENDIX I INDIVIDUAL BODY WEIGHTS (g) OF HALE MICE GROUP I - 0 PPM DAYS ON TEST: 0 7 ANIMAL 90620 90621 90622 90623 90624 26.4 27.3 27.1 26.7 26.2 30.4 31.0 31.5 28.5 28.3 14 31.5 32.6 33.4 30.8 30.3 GROUP III - DAYS ON TEST: 0 3 PPM 7 ANIMAL # 90625 90626 90627 90628 90629 28.2 26.4 29.0 25.3 24.4 30.7 31.6 32.1 28.3 30.2 14 31.6 32.2 33.6 30.1 32.2 GROUP V - 30 PPM DAYS ON TEST: 0 7 ANIMAL 90630 90631 90632 90633 90634 26.3 26.0 28.7 27.3 27.8 30.2 28.6 31.3 28.7 29.0 14 31.7 30.6 33.5 31.4 30.1 - 17 - Company Sanitized. Does nol conlam T5CA CBS - ^ifO^^tW I'--- DuPont HLR 325-95 REPEATED DOSE ORAL TOXICITY; 14-DAY FEEDING STUDY WITH H-19683 IN HALE MICE APPENDIX I (Continued) INDIVIDUAL BODY WEIGHTS (g) OF MALE MICE GROUP VII - 300 PPM DAYS ON TEST: 0 7 14 ANIMAL 90635 90636 90637 90638 90639 28.6 26.1 29.6 25.2 2".6 29.9 31.4 30.4 28.2 29.2 31.0 32.3 32.1 29.1 30.7 GROUP IX - 3000 PPH DAYS ON TEST; O 7 14 ANIMAL 90640 90641 90642 90643 90644 28.1 28.7 27.9 2'.! 2'. 3 27.5 26.7 22.1 27.0 28.1 28.6 28.3 26.7 28.4 31.4 18 Company Saniiized. Does notcorila1nTSCACB| DuPont HLR 325-95 APPENDIX II INDIVIDUAL ANIMAL FINAL BODY AND ORGAN WEIGHTS Explanatory Notes STD. DEV. = Standard Deviation - 19 - Company ffanffizerf. Does nof contain T5CA CBJ ^ DuPont HLR 325-95 REPEATED DOSE ORAL TOXICITYs 14-DAY FEEDING STUDY WITH H-19683 IM HALE MICE APPENDIX II INDIVIDUAL ANIMAL FIHAL BODY AND ORGAN WEIGHTS FINAL BODY AND ABSOLUTE ORGAN WEIGHTS (grans) FROM MALE MICE GROUP 0 PPM ANIMAL NUMBER FINAL BODY LIVER 90620 90621 90622 90623 90624 31.5 32.6 33.4 30.8 30.3 1.783 1.903 1.951 1.705 1.711 GROUP MEAN STD. DEV. 31.7 1.3 1.811 0.112 GROUP III PPM ANIMAL NUMBER FINAL BODY LIVER 90625 90626 90627 90628 90629 31.6 32.2 33.6 30.1 32.2 GROUP MEAN STD. DEV. 31.9 1.3 GROUP V - 30 PPM 2.098 1.914 1.946 1.906 1.669 1.907 0.154 ANIMAL NUMBER FINAL BODY LIVER 90630 90631 90632 90633 90634 GROUP MEAN STD. DEV. 31.7 30.6 33.5 31.4 30.1 31.5 1.3 2.370 1.909 2.736 2.265 1.924 2.241 0.344 20 ^Company Sanitized. Does not contain TSCA CBS DuPont P.-& 325-95 REPEATED DOSE ORAL TOXICITYs 14-DAY FEEDING STUDY WITH H-19683 IN MALE MICE GROUP VII - 300 PPM ANIMAL NUMBER FINAL BODY LIVER 9063J 90636 90637 90638 90639 GROUIP MEAN STD. DEV. 11.0 32.3 32.1 29.1 30.7 31.0 1.3 2.886 2.541 3.306 2.540 3.082 2.871 0.336 GROUP IX - 3000 PPM ANIMAL NUMBER 90640 90641 90642 90643 90644 GROlIP MEAN STD. DEV. FINAL BODY LIVER 28.6 28.3 26.7 28.4 31.4 28.7 1.7 4.343 5.014 3.294 4.266 4.909 4.365 0.685 - 21 Company Sanitized. Does not contain TSCA CB1 DuPont HLR 325-95 REPEATED DOSE ORAL TOXICITYs 14-DAY PE2DING STUDY rfITH H-196L3 IN MALE MICE APPENDIX II (continued) TWDTVIDUAL ANIMAL FINAL BODY ANO ORGAN WEIGHTS RELATIVE ORGftN WEIGHTS JX of body weight) FROM MALE MICE GROUP I - 0 PPM ANIMAL NUMBER LIVER 90620 90621 90622 90623 9062.4 5.6603 5.8374 5.8413 5.53^7 5.6469 GROUP MEAN STD. DEV. 5.7043 0.1324 GROUP III - ANIMAL NUMBER 3 PPM LIVER 90625 90626 90627 90628 90629 6.6392 5.9441 5.7917 6.3322 5.1832 GROUP MEAN STD. DEV. 5.9781 0.5546 GROUP V - 30 PPM ANIMAL NUMBER LIVER 90630 90631 90632 90633 90634 GROUP MEAN STD. DEV. 7.4763 6.2386 8.1672 7.2134 6.3920 7.0975 0.7963 - 22 - Company Sanitized. Does not contain T8CA CBI DuPont HLR 325-95 REPEATED DOSE ORAL TOXICITY: 14-DAY FEEDING STUDY VITH H-19683 IN MALE NICE APPENDIX II (continued) INDIVIDUAL ANIMAL FINAL BODY AND ORGAN WEIGHTS RELATIVE ORGAN WEIGHTS (X of body weight) FROM MALE MICE GROUP VII - 300 PPM ANIMAL NUMBER LIVER 90635 90636 90637 90638 90639 9.3097 7.8669 10.2991 8.7285 10.0391 C-FOUP MEAN S-3. DEV. 9.2486 0.9889 ;aoup ix - 3000 PPM ANIMAL NUMBER LIVER 90640 90641 90642 90643 90644 15.1853 17.7173 12.3371 15.0211 15.6338 G?:'UP MEAN 5~3. DEV. 15.1789 1.9204 - 23 - Cofftpany Sanitized. Does not contain TSCA CBI