Document KRogqqVRe7x7511Vvkz6BDyp0

Telom er Research Program AR226-1940 Report Title: 8-2 Telomer B Alcohol: Developmental Toxicity Study in Rats Author(s): Susan M. Munley, M.A. Contractor: E.I. du Pont de Nemours and Company, Haskell Laboratory for Health and Environmental Sciences, Elkton Road, P.O. Box 50. Newark, Delaware 19714-0050 Study Dates: January 30, 2003 (Study Initiation) - January 30, 2004 (Report Issuance) Study Objective The objective of this study was to evaluate the potential maternal and developmental toxicity of 8-2 Telomer B Alcohol when administered by oral gavage to pregnant rats daily from around the time of implantation to the end of gestation, days 6-20 of gestation (days 6-20G). Materials and Methods Groups of 22 time-mated Crl:CD(SD)IGS BR rats were administered formulations of the test substance in 0.5% aqueous methylcellulose over days 6 through 20 of gestation (G) at daily dose levels of 0, 50, 200, or 500 mg/kg. During the in-life portion of the study, body weight, food consumption, and clinical observation data were collected. On day 21G, all surviving dams were euthanized and dissected; the abdominal and thoracic viscera were examined grossly. The uterus of each pregnant rat was removed and dissected to permit examination of the uterine contents. The fetuses were removed, individually identified, weighed, sexed, and examined for external, visceral, head, and skeletal alterations. Findings (Report Summary Tables Attached) Under the conditions of this study, adverse maternal toxicity was produced at 500 mg/kg/day and consisted of maternal mortality, decreased body weights and body weight gains, and increased clinical observations of toxicity. One litter at 500 mg/kg/day consisted of one early resorption; persistent reductions in food consumption and body-weight gain and increased clinical observations indicative of toxicity were reported during the study for this female. Therefore, this litter consisting of a single early resorption is believed to be secondary to overt maternal toxicity in this dam. Developmental toxicity at 500 mg/kg/day consisted of increased skeletal variations (delayed pelvic bone ossification and wavy ribs). At 200 and 500 mg/kg/day, there were non adverse, test substance-related, transient reductions in maternal food consumption. In addition, there were slight increases in the incidence of delayed skull bone ossification at 200 and 500 mg/kg/day that were not considered adverse. Conclusion The no-observed-effect level (NOEL) for both maternal and developmental toxicity was considered 200 mg/kg/day. p Publications / Presentations: None Ol 2005 MAR -4 PM 12:01 Does not contain TSCA CBI 1 TRp (22-jan-04) Tables Explanatory Notes Notes Day C21 = Corrected final maternal body weight = final maternal body weight minus the gravid uterine weight plus the empty uterine weight. Abbreviations --= S.D. = S.E. = N= No Data Standard Deviation Standard Error Number in Group Does not contain TSCA CBI 2 TRP (22-Jan-04) TABLE 1 SUMMARY OF DOSING ANALYSES Nominal: Homogeneity Samples Test Day 1 Top Middle Bottom Dosing Concentration of 8-2 Telomer B Alcohol (mg/mL) 10 40 100 8.92 a (89.2) b 8.73 a (87.3) 10.0 (100.0) 40.5 (101.3) 40.0 (100.0) 39.8 (99.5) 96.5 (96.5) 98.3a (98.3) 94.8 (94.8) Average Measured Conc.c Average Percent Nominal Standard Deviationc Coefficient of Variationc 9.22 (92.2) 0.69 7% 40.1 (100.3) 0.37 1% 96.5 (96.5) 1.8 2% Stability Samples 5 Hour Room Temperature Concentration Verification Test Day 11 #1 #2 #3 9.56 (95.6) 7.28 a (72.8) 7.23 a (72.3) 8.71 d (87.1) 41.1 (102.8) 39.9 a (99.8) 39.7 a (99.3) 41.1 d (102.6) 104 (104.0) 100a (100.0) 101 a (101.0) 98.2d (98.2) Average Measured Conc. e Average Percent Nominal e Standard Deviation e Coefficient of Variation e Test Day 20 #1 7.74 (77.4) 0.84 11% 9.45 (94.5) 40.2 (100.5) 0.76 2% 35.0 (87.5) 99.7 (99.7) 1.4 1% 94.6 (94.6) #2 9.57 37.8 91.9 (95.7) (94.5) (91.9) Average Measured Conc.f Average Percent Nominalf Standard Deviation f Coefficient of Variation1 9.51 (95.1) 0.09 1% 36.4 (91.0) 2.0 5% 93.2 (93.2) 1.9 2% a Mean result of the original analysis and duplicate reanalysis of original sample. b Numbers in parentheses are the respective percent of nominal values. c Statistics based on the average measured concentration of the top, middle and bottom of each dosing level. d Result of the analysis for the third sample collected from the dosing preparations. e Statistics based on the average measured concentration of triplicate samples for the dosing level. f Statistics based on the average measured concentration of duplicate samples for the dosing level. Does not contain TSCA CBI 3 TRP (22-Jan-04) TABLE 2 CLINICAL OBSERVATIONS DAY OF GESTATION 6-21 OBSERVATION: GROUP : DOSE RATE (MG/KG/DAY): NO. EXAMINED : NO. SACRIFICED IN EXTREMIS NO. FOUND DEAD ALOPECIA DIARRHEA HUNCHED OVER PALLOR SORE STAIN CAGEBOARD STAIN FACE STAIN PERINEUM WEAK WET PERINEUM I 0 22 0 0 5 0 0 0 0 0 0 0 0 0 * Statistically significant trend (Cochran-Armitage test p < 0.05. II 50 22 0 0 2 0 0 0 0 0 0 0 0 0 III IV 200 500 22 22 0 4* 0i 6 10* 0 6* 0 6* 01 01 11 01 0 3* 0 5* 01 Does not contain TSCA CBI 4 TRP (22-Jan-04) TABLE 3 MEAN MATERNAL BODY WEIGHTS (grams)a DAY 0 GROUP I: 0 MG/KG/DAY GROUP MEAN S.D. S.E. N 249.4 10.26 2.19 22 DAY 4 265.7 11.68 2.49 22 DAYS OF GESTATION DAY 5 DAY 6 DAY 7 270.2 11.37 2.42 22 277.5 12.33 2.63 22 284.5 11.77 2.51 22 DAY 8 289.9 11.88 2.53 22 DAY 9 293.9 11.65 2.48 22 GROUP II: 50 MG/KG/DAY GROUP MEAN S.D. S.E. N 249.6 11.52 2.51 21 264.6 11.61 2.53 21 271.0 12.86 2.81 21 276.3 12.99 2.84 21 281.5 12.38 2.70 21 287.9 11.15 2.43 21 291.6 10.33 2.26 21 GROUP III: 200 MG/KG/DAY GROUP MEAN S.D. S.E. N 249.8 11.38 2.43 22 259.1 16.20 3.45 22 269.2 13.81 2.94 22 274.6 13.09 2.79 22 278.2 12.21 2.60 22 284.8 13.59 2.90 22 287.7 11.90 2.54 22 GROUP IV: 500 MG/KG/DAY GROUP MEAN S.D. S.E. N 249.7 12.02 3.00 16 263.5 16.57 4.14 16 269.8 16.81 4.20 16 277.8 16.17 4.04 16 271.9* 19.90 4.97 16 278.3* 20.79 5.20 16 284.2* 22.40 5.60 16 Does not contain TSCA CBI 5 TRP (22-Jan-04) TABLE 3 (Continued) MEAN MATERNAL BODY WEIGHTS (grams) DAY 10 GROUP I: 0 MG/KG/DAY GROUP MEAN S.D. S.E. N 299.6 12.50 2.67 22 DAY 11 306.8 14.74 3.14 22 DAYS OF GESTATION DAY 12 DAY 13 DAY 14 314.3 14.58 3.11 22 322.4 13.49 2.88 22 326.0 15.44 3.29 22 DAY 15 332.8 15.39 3.28 22 DAY 16 344.3 16.87 3.60 22 GROUP II: 50 MG/KG/DAY GROUP MEAN S.D. S.E. N 297.1 11.84 2.58 21 304.3 11.02 2.40 21 311.8 11.99 2.62 21 318.5 10.86 2.37 21 324.1 12.77 2.79 21 329.5 10.78 2.35 21 340.1 11.37 2.48 21 GROUP III: 200 MG/KG/DAY GROUP MEAN S.D. S.E. N 292.5 11.55 2.46 22 299.3 12.41 2.65 22 308.1 14.38 3.07 22 316.7 15.51 3.31 22 320.7 16.25 3.46 22 330.2 18.81 4.01 22 341.5 19.07 4.07 22 GROUP IV: 500 MG/KG/DAY GROUP MEAN S.D. S.E. N 287.8* 17.89 4.47 16 286.8* 21.52 5.38 16 296.0* 25.14 6.29 16 303.7* 23.68 5.92 16 313.2* 20.46 5.12 16 319.8* 23.03 5.76 16 331.2 23.55 5.89 16 Does not contain TSCA CBI 6 TRP (22-Jan-04) TABLE 3 (Continued) MEAN MATERNAL BODY WEIGHTS (grams) DAY 17 GROUP I: 0 MG/KG/DAY GROUP MEAN S.D. S.E. N 358.2 17.52 3.74 22 DAY 18 374.5 18.14 3.87 22 DAYS OF GESTATION DAY 19 DAY 20 DAY 21 DAY C21 391.1 20.22 4.31 22 409.0 21.87 4.66 22 433.8 25.95 5.53 22 330.3 20.07 4.28 22 GROUP II: 50 MG/KG/DAY GROUP MEAN S.D. S.E. N 354.1 12.58 2.74 21 369.5 13.83 3.02 21 385.2 14.86 3.24 21 401.6 15.38 3.36 21 427.1 18.14 3.96 21 332.4 14.87 3.24 21 GROUP III: 200 MG/KG/DAY GROUP MEAN S.D. S.E. N 356.8 21.47 4.58 22 373.2 23.80 5.07 22 389.2 26.02 5.55 22 405.3 29.39 6.27 22 426.5 32.75 6.98 22 325.7 23.20 4.95 22 GROUP IV: 500 MG/KG/DAY GROUP MEAN S.D. S.E. N 346.1 25.61 6.40 16 360.5 27.43 6.86 16 374.1* 28.59 7.15 16 386.2* 28.17 7.04 16 395.1* 34.57 8.64 16 300.4 29.09 7.27 16 a Data were excluded from females that were not pregnant, found dead, sacrificed in extremis, or had litters consisting entirely of resorptions. * Statistically significant trend (linear contrast of means); p < 0.05. Does not contain TSCA CBI 7 TRP (22-Jan-04) TABLE 4 MEAN MATERNAL BODY WEIGHT CHANGES (grams) a 0-6 GROUP I: 0 MG/KG/DAY GROUP MEAN S.D. S.E. N 28.1 8.30 1.77 22 DAYS OF GESTATION 4-6 6-8 8-10 10-12 12-14 14-16 11.8 3.75 0.80 22 12.4 4.74 1.01 22 9.7 4.60 0.98 22 14.7 5.79 1.23 22 11.8 4.38 0.93 22 18.2 4.48 0.96 22 GROUP II: 50 MG/KG/DAY GROUP MEAN S.D. S.E. N 26.7 6.12 1.33 21 11.7 3.67 0.80 21 11.5 5.27 1.15 21 9.2 4.23 0.92 21 14.8 6.61 1.44 21 12.3 5.72 1.25 21 16.0 7.95 1.74 21 GROUP III: 200 MG/KG/DAY GROUP MEAN S.D. S.E. N 24.8 5.77 1.23 22 15.4 7.95 1.70 22 10.2 6.47 1.38 22 7.7 5.14 1.10 22 15.6 5.52 1.18 22 12.6 6.31 1.35 22 20.8 5.71 1.22 22 GROUP IV: 500 MG/KG/DAY GROUP MEAN S.D. S.E. N 28.1 7.99 2.00 16 14.2 5.09 1.27 16 0.5* 9.68 2.42 16 9.5 9.26 2.31 16 8.2* 14.95 3.74 16 17.2* 10.08 2.52 16 18.0 8.65 2.16 16 Does not contain TSCA CBI 8 TRP (22-Jan-04) TABLE 4 (Continued) MEAN MATERNAL BODY WEIGHT CHANGES (grams) 16-18 GROUP I: 0 MG/KG/DAY GROUP MEAN S.D. S.E. N 30.3 4.18 0.89 22 DAYS OF GESTATION 18-21 6-21 6-C21 59.3 9.91 2.11 22 156.4 21.46 4.58 22 52.9 16.15 3.44 22 GROUP II: 50 MG/KG/DAY GROUP MEAN S.D. S.E. N 29.4 6.63 1.45 21 57.6 7.96 1.74 21 150.8 19.57 4.27 21 56.1 13.63 2.97 21 GROUP III: 200 MG/KG/DAY GROUP MEAN S.D. S.E. N 31.8 7.02 1.50 22 53.3* 11.41 2.43 22 151.9 28.17 6.01 22 51.1 19.49 4.16 22 GROUP IV: 500 MG/KG/DAY GROUP MEAN S.D. S.E. N 29.3 7.50 1.88 16 34.6* 24.96 6.24 16 117.4* 34.43 8.61 16 22.6* 30.68 7.67 16 a Data were excluded from females that were not pregnant, found dead, sacrificed in extremis, or had litters consisting entirely of resorptions. * Statistically significant trend (linear contrast of means); p < 0.05. Does not contain TSCA CBI 9 TRP (22-Jan-04) TABLE 5 MEAN MATERNAL FOOD CONSUMPTION (grams/day) a 4-6 GROUP I: 0 MG/KG/DAY GROUP MEAN S.D. S.E. N 22.8 2.78 0.59 22 DAYS OF GESTATION 8-10 10-12 12-14 14-16 16-18 CO 23.3 3.47 0.74 22 24.1 2.35 0.50 22 25.5 2.94 0.63 22 26.1 2.68 0.57 22 27.2 2.65 0.56 22 29.3 3.83 0.82 22 GROUP II: 50 MG/KG/DAY GROUP MEAN S.D. S.E. N 23.8 2.56 0.56 21 21.9 2.42 0.53 21 23.4 2.49 0.54 21 25.7 2.71 0.59 21 26.4 2.29 0.50 21 26.8 3.68 0.80 21 30.2 3.42 0.75 21 GROUP III: 200 MG/KG/DAY GROUP MEAN S.D. S.E. N 23.5 2.39 0.51 22 21.3* 2.40 0.51 22 22.1* 2.22 0.47 22 24.6 3.65 0.78 22 26.1 3.57 0.76 22 28.4 3.28 0.70 22 31.9* 2.96 0.63 22 GROUP IV: 500 MG/KG/DAY GROUP MEAN S.D. S.E. N 23.7 3.03 0.76 16 19.9* 5.91 1.48 16 21.4* 3.25 0.81 16 20.6* 6.76 1.69 16 25.8 4.28 1.07 16 27.9 4.44 1.11 16 31.8* 5.31 1.33 16 Does not contain TSCA CBI 10 TRP (22-Jan-04) TABLE 5 (Continued) MEAN MATERNAL FOOD CONSUMPTION (grams/day) DAYS OF GESTATION 18-21 6-21 GROUP I: 0 MG/KG/DAY GROUP MEAN S.D. S.E. N 28.1 3.84 0.82 22 26.4 2.64 0.56 22 GROUP II: 50 MG/KG/DAY GROUP MEAN S.D. S.E. N 28.2 2.47 0.54 21 26.2 2.00 0.44 21 GROUP III: 200 MG/KG/DAY GROUP MEAN S.D. S.E. N 28.4 3.88 0.83 22 26.3 2.46 0.52 22 GROUP IV: 500 MG/KG/DAY GROUP MEAN S.D. S.E. N 25.4 5.55 1.39 16 24.7 2.88 0.72 16 a Data were excluded from females that were not pregnant, found dead, sacrificed in extremis, or had litters consisting entirely of resorptions. * Statistically significant trend (linear contrast of means); p < 0.05. Does not contain TSCA CBI 11 TRP (22-Jan-04) TABLE 6 REPRODUCTIVE OUTCOME Group: Dose (mg/kg/day): I 0 II 50 No. Mated No. Pregnant No Delivered Early No. Deaths No. With Total Resorptions No. Litters Mean Corpora Luteaa 22 22 0 0 0 22 15.7 (1.61)b 22 21 0 0 0 21 16.0 (3.04) III 200 22 22 0 0 0 22 16.0 (3.40) IV 500 22 22 0 5* 1 16 16.1 (2.51) Implantations (Nidations) Resorptions: Total Early Late Dead Fetuses Live Fetusesc : Total Males Females Mean Fetal Weight: Total Males Females 14.7 (1.62) 0.0 (0.21) 0.0 (0.21) 0.0 0.0 14.6 (1.56) 7.2 (1.79) 7.5 (1.74) 5.55 (0.31) 5.68 (0.33) 5.43 (0.32) 13.9 (1.56) 0.7 (1.06) 0.7 (1.06) 0.0 0.0 13.2 (1.81) 6.0 (1.58) 7.2 (1.69) 5.66 (0.29) 5.83 (0.34) 5.51 (0.26) 13.9 (1.64) 0.4 (0.79) 0.4 (0.79) 0.0 0.0 13.5 (1.77) 6.7 (2.16) 6.8 (1.97) 5.77 (0.39) 5.94 (0.40) 5.59 (0.42) 13.4 (3.87) 0.4 (0.81) 0.4 (0.81) 0.1 (0.25) 0 .0 13.8 (1.95) 7.1 (1.84) 6.7 (1.89) 5.40 (0.44) 5.57 (0.47) 5.20 (0.39) Sex Ratiod 0.49 (0.11) 0.45 (0.11) 0.50 (0.14) 0.51 (0.12) a Statistical analyses are not conducted on mean corpora lutea data; these data are presented for information only. b Standard deviation is reported in parentheses. c Statistical analyses are only conducted on the mean total number of live fetuses per litter and total mean fetal weight. The means for males and females are presented for information only. d Number male fetuses/total number fetuses per litter. Note: The pregnancy rate data, adult mortality data, and the total resorption data were statistically analyzed using the Cochran-Armitage test. All litter mean data (except for fetal weight and sex ratio) were analyzed using Jonckheere's test. Fetal weight and sex ratio were analyzed using a linear contrast of least square means.* * Statistically significant trend; p < 0.05. Does not contain TSCA CBI 12 TRP (22-Jan-04) EXTERNAL No. Examineda No. Affected TABLE 7 INCIDENCE OF FETAL MALFORMATIONS Group : Dose (mg/kg/day): I 0 II III 50 200 IV 500 322 [22] 0[ 0] 277 [21] 0[ 0] 297 [22] 0[ 0] 220 [16 0[ 0] VISCERAL No. Examined No. Affected 167 [22] 0[ 0] 142 [21] 0[ 0] 156 [22] 0[ 0] 113 [16 0[ 0] HEAD No. Examined No. Affected 167 [22] 0[ 0] 142 [21] 0[ 0] 156 [22] 0[ 0] 113 [16 0[ 0] SKELETAL N o . Examined No. Affected 322 [22] 0[ 0] 277 [21] 0[ 0] 297 [22] 0[ 0] 220 [16 0[ 0] TOTAL NUMBER AFFECTED 0( 0) 0( 0) 0( 0) 0( 0) a Number examined and affected, including the number affected with the listed malformations are expressed as Fetuses[Litters] or Fetuses (Litters). b For ease of reading, zeros have been replaced with dashes for the listed malformations. No significant trends were detected (Jonckheere-Terpstra Trend test); p < 0.05. Note: Statistical analyses are only conducted on the individual endpoints. The overall total and totals by exam type are presented for information only. Does not contain TSCA CBI 13 TRP (22-Jan-04) TABLE 8 INCIDENCE OF FETAL VARIATIONS Group: Dose (mg/kg/day): I 0 II III 50 200 DEVELOPMENTAL VARIATIONS EXTERNAL No. Examineda No. Affected 322 [22] 0[ 0] 277 [21] 0[ 0] 297 [22] 0[ 0] VISCERAL No. Examined No. Affected 167 [22] 0[ 0] 142 [21] 0[ 0] 156 [22] 0[ 0] HEAD No. Examined No. Affected 167 [22] 0[ 0] 142 [21] 0[ 0] 156 [22] 0[ 0] SKELETAL No. Examined Rib - Rudimentary Cervical - Supernumerary Skull - Fused No. Affected 322 [22] 277 [21] 297 [22] -- 25 (11) -- 20 (13) 2( 1) 32 (15) 25 (11) 20 (13) 34 (16) TOTAL WITH DEVELOPMENTAL VARIATIONS 25(11) 20(13) 34(16) VARIATIONS DUE TO RETARDED DEVELOPMENT EXTERNAL No. Examined3 No. Affected VISCERAL No. Examined Kidneyb Small Papilla - Size 1 Small Papilla - Size 2 Papilla - Size 3 No. Affected 322 [22] 0[ 0] 277 [21] 0[ 0] 297 [22] 0[ 0] 167 [22] 142 [21] 156 [22] 2( 2) 4( 3) 6( 6) 12[ 9] -- 2( 2) 5( 5) 7[ 7] 1( 1) 1( 1) 3( 3) 5[ 5] IV 500 220 [16 0[ 0] 113 [16 0[ 0] 113 [16 0[ 0] 220 [16 1( 1) 34 (10) 1( 1) 35 (11) 35(11) 220 [16 0[ 0] 113 [16 2( 2) 1( 1) 2( 1) 5[ 3] Does not contain TSCA CBI 14 TRP (22-Jan-04) TABLE 8 (Continued) INCIDENCE OF FETAL VARIATIONS VARIATIONS DUE TO RETARDED DEVELOPMENT (Continued) HEAD No. Examined 167 [22] No. Affected 0[ 0] 142 [21] 0[ 0] 156 [22] 0[ 0] 113 [16] 0[ 0] SKELETAL N o . Examined Pelvis - Retarded Ossification Rib - Wavy Skull - Retarded Ossification Sternebra - Retarded Ossification Vertebra - Retarded Ossification No. Affected 322 [22] 1( 1) --22(: 8) 5( 3) 3 7(:i2) 56( [15] 277 [21] ----- 31(12) 2( 2) 30( 8) 56[13] 297 [22] ----- 45(16)* 3( 2) 34(13) 72[19] 220 [16] 9( 5)* 5( 2)* 55(12)* 8( 5) 12( 6) 73[14] TOTAL WITH VARIATIONS DUE TO RETARDED DEVELOPMENT 66(:i7) 63(15) 76(19) 78(15) TOTAL NUMBER FETUSES WITH VARIATIONS 89( 21) 80(19) 99(20) 106(15) a Number examined and affected, including the number affected with the listed variations, are expressed as Fetuses[Litters] or Fetuses (Litters). b Statistical analyses are conducted on the combined incidences of small renal papillae; the details for each size are presented for information only. * Significant trend (Jonckheere's test); p < 0.05. Note: Statistical analyses are only conducted on the individual endpoints. overall total and totals by exam are presented for information only. The Does not contain TSCA CBI 15 TRP (22-Jan-04)