Document rxRQp64LXQ2XXNwd3Xz6mr6de

Telomer Research Program JU X AiA-aqt.ASS CI a ri a n t Report Title: 8-2 Telomer B Alcohol: Oral Gavage Range-Finding Study in Rats Author(s): Gregory S. Ladies, Ph.D ro 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 /V c1--z' 1 --1 Study Dates: Study Initiated: April 2001 Study Completed: September 2001 rsc w* an 10 Study Objective: The objective of this study was to set dose levels for a 90-day subchronic toxicity study with 8-2 Telomer B Alcohol. In addition, analytical methods were developed for looking at potential metabolites of 8-2 Telomer B Alcohol. 0^0 T>m ~x3<r> Om oO Materials and Methods: Young adult male (10/group) and female (5/group) Crl:CDR(SD)IGS BR rats were administered 8-2 Telomer B Alcohol daily by gavage at dosages of 0, 5, 25, and 125 mg/kg/day until the attainment of steady state blood fluorine levels, and allowed to recover for 75 days post-dosing (5/group, males and females). Serial bleeding for analysis of total fluorine and parent/metabolites was conducted during the dosing and recovery phases of the study. Blood fluorine levels reached steady state between test days 60-84. At steady state blood fluorine levels, the liver and kidneys of 5 males/group were weighed; females were not evaluated. At necropsy, liver kidney, and fat were saved from males and females for analysis of parent compound, PFOA, and PFNA. In addition, pooled plasma samples obtained from males and females over the duration of the in-life phase of the study were analyzed for parent compound, the 8-2 acid, PFOA, and PFNA. At steady state (test day 84) pooled plasma were analyzed for PFOA, parent compound, and total fluorine levels. Findings: In-Life Toxicology Parameters'. No test substance-related mortality occurred in the study. A test substance-related, toxicologically significant increase in the incidence of striated teeth was observed in the 125 mg/kg/day male and female dose groups. Mean final body weights and body weight gains were decreased in male rats in the 125 mg/kg/day dose group on test day 81. Body weights were similar to control at the end of the recovery period. Females body weights and body weight gains were not affected. Anatomical Pathology: Test-substance related and statistically significant increases in liver weight parameters occurred in males administered 125 mg/kg/day for 81 days. After 75 days of recovery, male rats dosed with 125 mg/kg/day showed reversibility of the organ weight effects. Does not contain TSCA CBI TRP (28-Apr-03) Female rats were not evaluated on test day 81. After 75 days of recovery, test-substance related and statistically significant increases in liver and kidney weight parameters occurred in females administered 125 mg/kg/day. Mean Final Body and Organ Weights for Male Rats Sacrificed on Test Day 81 Liver (g) Kidneys(g) Final body weight (g) 8-2 Telomer B Alcohol (mg/kg/day) 05 25 125 23.4 21.1 3.4 2.5 22.7 2.1 27.9 + 3.1 4.5 0.47 4.3 0.34 4.1 0.31 4.4 0.68 594.1 564.1 44.35 59.08 552.6 33.89 508.9 54.71# Liver (% BW) 3.9 0.28 3.7 + 0.20 Kidneys (% BW) 0.76 0.03 0.76 0.06 Mean + standard deviation. The n = 5. # Statistically significant difference from control (p = 0.05). 4.1 0.19 0.75 0.04 5.5 0.39# 0.87 0.11 Does not contain TSCA CBI 2 TRP (28-Apr-03) Mean Final Body and Organ Weights for Male Rats Sacrificed on Post Dose Day 75 Liver (g) Kidneys(g) Testes (g) Final body weight (g) 8-2 Telomer B Alcohol (mg/kg/day) 0 5 25 125 16.9 3.2 18.7 2.6 16.9 + 2.2 17.2 2.2 3.6 0.45 3.9 0.32 3.9 0.47 4.0 0.39 3.6 + 0.28 3.5 0.55 3.6 0.34 3.4 0.74 553.5 594.7 573.5 548.9 99.82 57.07 83.19 36.78 Liver (% BW) Kidneys (% BW) 3.0 0.13 0.65 3.1 0.14 0.67 + 2.9 + 0.13 0.69 3.1 0.32 0.74 Testes (% BW) 0.7 0.10 Mean + standard deviation. The n = 5. # Statistically significant difference from control (p = 0.05). 0.6 0.08 0.6 0.09 0.6 0.11 Mean Final Body and Organ Weights for Female Rats Sacrificed on Post Dose Day 75 Liver (g) Kidneys(g) Final body weight (g) 8-2 Telomer B Alcohol (mg/kg/day) 0 5 25a 125 10.3 1.7 11.1 1.3 11.3 2.5 12.7 1.2 2.2 0.21 2.4 0.04 2.4 0.37 2.7 0.17# 322.2 348.1 332.1 345.5 41.03 29.21 50.42 31.34 Liver (% BW) 3.2 0.14 3.2 0.33 3.4 0.37 Kidneys (% BW) 0.70 0.03 0.71 0.07 0.73 0.06 Mean standard deviation. The n = 4 unless otherwise noted; The n = 5. # Statistically significant difference from control (p = 0.05). 3.6 + 0.05# 0.78 0.02# Does not contain TSCA CBI 3 TRP (28-Apr-03) ppm F in blood Blood fluorine analyses: Blood fluorine levels reached steady state between test days 60-84. Total Blood Fluorine Analysis in Male Rats Dosed with Telomer 8-2 Alcohol Total Blood Fluorine Analysis In Female Rats Dosed with Telomer 8-2 Alcohol Does not contain TSCA CBI TRP (28-Apr-03) r Analysis o f 8-2 Telomer B Alcohol, PFOA, and PFNA_ in Serum and Selected Tissues: Pooled plasma samples obtained from males and females at steady state (test day 84) were analyzed for PFOA, parent compound, and total fluorine levels. The level of parent detected did not exceed 0.074 ppm in either males or females. The following plasma levels of PFOA were detected in males and females dosed with 0, 5, 25, and 125 mg/kg/day 8-2 Telomer B Alcohol. Dose Group (mg/kg/day) Male PFOA (ppm) Male PFOA converted to ppm F Male total plasma F (ppm)a Female PFOA (ppm) Female PFOA converted to ppm F Female total plasma F (ppm)a 0 0.07 0.05 NM 0.0 0.0 NM 5 2.19 1.51 NM 0.08 0.05 NM 25 8.47* 5.83 8.11 0.54* 0.37 5.88 125 65.60* 45.16 53.24 1.85* 1.27 23.20 `'Determined using Wickbolc Torch; *mean of two separate runs; NIV - not measured Based on the data presented above, the PFOA in males accounts for a large portion of the fluorine present in the plasma, however, this is not the case for females. While what is in the blood in males is predominately PFOA, it is not possible to detennine what percentage of the 8-2 Telomer B Alcohol dose is converted to PFOA (e.g., information is lacking on mass balance and biodistribution). Further, it is less certain as to what the metabolites may be in females as both parent and PFOA were present in low levels and did not account for the total fluorine levels in the blood. Tissue samples (fat, liver, and kidney) obtained at necropsy from males and females at steady state (test day 81) were analyzed for the concentration of parent compound, PFOA, and PFNA. The data are summarized below. Tissue Analyses from Range-Finder Study (Test Day 81) 8-2 Telomer B Alcohol (mg/kg/day) 5 25 125 C8-2A (ppm) Fat Liver Kidney 0.07 (0.01) 0.02 (0.00) ND 0.34 (0.13) 0.13 (0.03) 0.01 (0.00) 1.37 (0.55) 0.77 (0.53) 0.07 (0.07) PFOA (ppm) Plasma Fat Liver Kidney 2.19 0.09 (0.04) 6.96 (1.94) 1.64 (0.83) 8.47 0.35 (0.12) 18.68 (10.38) 8.43 (4.14) 65.60 1.90 (1.66) 30.69 (20.68) 29.11 (17.38) PFNA (ppm) Fat Liver Kidney 0.00 (0.00) 0.51 (0.09) 0.00 (0.00) 0.00 (0.00) 0.91 (0.23) 0.11 (0.09) 0.01 (0.01) 1.19(1.25) 0.86 (0.62) Does not contain TSCA CBI 5 Q TRP (28-Apr-03) Serum samples were collected throughout the in-life phase of the study and pooled within each dose group (per sex) for analysis of parent compound, PFOA, PFNA, and the 8-2 acid (CioHiChFn). Since there is no standard available for the 8-2 acid, we are unable to quantitate or positively identify this species in the plasma (fat, liver and kidney were not analyzed). However, there is an unknown at mass/charge ratio of 477, which is consistent with the 8-2 acid. This 477 species appears to have a dose response relationship, further supporting the premise that this species is related to the metabolism of the 8-2 alcohol. Since it is not possible to quantitate, we have presented the data as area counts rather than absolute concentrations. Although we cannot quantitate, if we assume that the response factor for this unknown metabolite is approximately equal to the response factor for PFNA, then the concentrations would be in the low ppm range (i.e. more similar to PFNA than PFOA). The data are presented below. Does not contain TSCA CB1 6 7 TRP (28-Apr-03) Plasma PFOA Concentration in Male Rats Plasma PFOA Concentration in Female Rats -0 mg/kg/day -5 mg/kg/day -25 mg/kg/day -125 mg/kg/day Does not contain TSCA CBI 7 TRP (28-Apr-03) P la s m a 8-2 T e lo m e r B A Ic o h o l C o n c e n tra tio n in M a le Rats 5 mg/kg/day 25 mg/kg/day 125 mg/kg/day Does not contain TSCA CBI 8 TRP (28-Apr-03) P la s m a PFNA C o n c e n tra tio n in M ale Rats - 0 mg/kg/day ----- 5 mg/kg/day - - - 25 mg/kg/day -12 5 mg/kg/day 2.00 n _ 1.50 - EO) 3. 1.00 - L 0.00 - 0 Plasma PFNA Concentration in Female Rats - 0 mg/kg/day - 5 mg/kg/day -- r ~ - 25 mg/kg/day -- X r ~ -1 2 5 mg/kg/day Test day Does not contain TSCA CBI 9 /o TRP (28-Apr-03) 2 0 0 0 0 0 0 0 -, 18000000 16000000 - 14000000 - 12000000 - 2 10000000 - < 8000000 6000000 - 4000000 - 2000000 - 00 P la s m a M e t4 7 7 C o n c e n tra tio n in M a le Rats Test day ----- Series2 --A--- Series3 X - Series4 P lasm a M et4 77 C o n c e n tra tio n in F em ale Rats 2 0 0 0 0 0 0 0 -, 18000000 16000000 14000000 12000000 2 10000000 < 8000000 - 6000000 4000000 2000000 - 0- 0 7 14 21 28 35 42 49 56 63 70 77 Test day m-- 5 mg/kg/day -- 25 mg/kg/day -- 125 mg/kg/day Does not contain TSCA CBI 10 1/ TRP (28-Apr-03) Conclusion: Based on the data from this range-finder study, dose levels of 1, 5, 25, and 125 mg/kg/day were selected for the 90-day subchronic toxicity study. Publications / Presentations: None. Does not contain TSCA CBI 11 TRP (28-Apr-03)