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Study NO.T-7098.1; DT35 Title: &zu 4 69 5 Pharmacokinetic Study of Perfluorooctanesulfonyl Fluoride {POSF) FollowinE a Single Oral Gavage Dose In Rats 3M Strategic Toxicology Laboratory Corporate Toxicology 3M Medical Department 3M Center, Building 270-SB-3 14 St. Paul, MN 55144 Final Report January 31,2004 Study Director: Andrew M. Seacat, Ph.D., DABT, Toxicology Specialist Study Toxicologist: Deanna J. Luebker, M.S., Advanced Research Toxicologist Sponsor: 3M Specialty Chemicals Division 3M Center Bldg 236 St. Paul, MN 55144 Page 1 of 11 T-7098.1; ST35 Final Report Summary Rats received a single oral dose of 5 mg/Kg perfluorooctanesulfonyl fluoride. Necropsies were - performed on days 1,4, and 29 post dose. POSF was metabolized to PFOS which was maximum in the liver on day 4 post-dose at approximately 11 ppm, or 10% of the dose. PFOS was present in sera at approximately one tenth of the level found in the liver. Thus, perfluorooctanesulfonyl fluoride (POSF) was absorbed and metabolized to perfluorooctanesulfonate (PFOS) by rats following and oral dose. Study Objective The objective of this study was to assess the potential for oral absorption, urinary and fecal clearance, and biological persistence of perfluorooctanesulfonyl fluoride (POSF) in male Sprague Dawley rats after a single oral dose. POSF is the starting material for the synthesis of a variety of perfluorooctane sulfonate (PFOS) based materials. Data gathered in this study help define the rate of metabolism of POSF to PFOS by the liver and provide data for proper risk characterization of POSF. Method Summary This study was performed in the 3M Strategic Toxicology Laboratory under a defined protocol (1) and classified as a "Class B Study" as explained in TOX SOP 0950, Strategic Toxicology Lab GLP Program Procedure (2). Briefly, the procedure was as follows. Thirty male Sprague Dawley rats (obtained from Harlan Laboratories), 6-8 weeks in age and approximately 150-250g, were divided into the following dose groups: Euthanasia day 1 post dose day 4 post dose day 29 post dose day 1 post dose day 4 post dose day 29 post dose Rats received either a 5 mlKg dose of vehicle control (2% Tween SO), or a 5 mLKg dose of a 1 mg/mL POSF suspension in 2% Tween 80 (final dose = 5 mgiKg) via oral gavage on day zero of the study. Three specified rats from groups 3 and 6 were housed individually in metabolism cages for portions of the study. Urine and feces were collected on days 1,2,4, 14, and 29-post dose. Necropsies were performed on days 1,4, and 29 post dose. At necropsy, liver and sera were collected from each animal. For metabolite analysis, these specimens were packed in dry ice and shipped to Kris Hansen, 3M Environmental Technology and Safety Services for FC 3 Page 2 of 11 j T-7098.1; ST35 Final Report analysis. Liver and sera were analyzed and the raw data was reported (3) . Urine and feces analysis was not performed and was canceled. Results Treatment with PFOS at 5 m a g had no effect on body weight or liver weight (Table lA, B). Extraction and analysis of the tissues showed that PFOS was detected in the livers of all POSF dosed animals at all time points and was maximum in the liver on day 4 post-dose at approximately 11 ppm (Table 2). Trace amounts of perfluorooctanesulfonamidoacetate (PFOSAA, M556) were also reported for liver samples from POSF treated animals on days one and 4 post-dose (Table 2). The origin of the M556 in the liver samples is unknown, and all samples were near the practical quantitation limit for PFOSAA of 0.06 pg/g. Approximately 10 % of the dose was present in the liver as PFOS on day 4 post dose as calculated fiom the sum of the liver PFOS and liver PFOS equivalents from PFOSAA (Table 3). The PFOS levels in the sera of the rats were approximately one tenth of the level found in the liver sample extracts and were maximal on day one post dose (Table 4). No other metabolites were detected in the sera. Approximately 0.1 % of the dose was present in the sera as PFOS on day 1 post-dose (Table 4). Conclusions These data support the conclusion that perfluorooctanesulfonyl fluoride (POSF) can be absorbed by rats following a single oral gavage dose, and metabolized to perfluorooctanesulfonate (PFOS). , Page 3 of 11 4 L T-7098.1; ST35 Final Report k. List of Tables 1. Table 1: Biological Parameters 1A: Control Group 1B: POSF Dose Group. 2. Table 2: Liver PFOS and PFOSAA (M556) 3. Table 3: Liver PFOS equivalents and Percent Dose in Liver 4. ;Table4: Serum PFOS and Percent Dose in serum Page 4 of 11 \., T-7098.1; ST35 Final Report Table 1A: Biological Parameters. Control Group Time Dose Point animal # Initial Terminal Kidney Liver wt 9ROO-xxx BW (g) BW (g) Weight (g) (g) Day1 Omgkg 00 1 202 207 1.7 8.7 Day1 Day1 Day1 Day1 Omgkg Omgkg Omgkg Omgkg 002 204 213 003 188 196 004 203 207 005 200 207 1.7 9.3 1.6 8.4 1.7 9.5 1.6 9 Avg SD Range Min MU N 199 206 6 6 188 196 204 213 5 5 5 1.7 9.0 0.1 0.4 1.6 8 1.7 10 5 5 Day4 Day4 Day4 Day4 Day4 Omgkg Omgkg Omgkg Omg/kg Omgkg 006 193 206 007 198 215 008 204 228 009 204 220 010 191 209 1.6 8.2 1.7 8.6 1.8 9.5 1.7 9.1 1.7 9.1 Avg SD Range Min Max N 198 215 6 9 191 206 204 228 5 5 5 1.7 9 0.1 1 1.6 8 1.8 10 5 5 Day29 Omg/kg 01 1 196 275 2.1 11.9 Page 5 of 11 T-7098.1;ST35 Final Report Table 1: 'Table 1B: Biological Parameters. POSF Dose Group /Time Point Dose animal ## Initial Terminal 9ROO-xxx BW (8) BW (g) Kidney Weight (8) > Liver wt (g) Day1 Day1 Day1 Day1 Day1 5mg/kg 5mgkg 5mg/kg 5mg/kg 5mgkg Avg 03 1 21 1 215 032 199 203 033 204 213 034 197 205 03 5 212 216 205 210 1.6 10.0 1.6 8.7 1.9 9.8 1.5 8.7 1.7 8.6 1.7 9 ~ SD Range Min Max N 7 6 197 203 212 216 5 5 5 0.2 1 1.5 9 1.9 10 5 5 Day4 Day4 Day4 Day4 Day4 5mgkg 5mgkg 5mg/kg 5mg/kg 5mg/kg Avg SD Range N Min Max 036 215 240 037 195 216 038 200 22 1 039 203 229 040 199 224 202 226 7 9 195 216 215 240 5 5 5 1.9 10.2 1.7 9.0 1.7 9.3 1.9 9.8 1.7 9.3 1.8 10 0.1 0 1.7 9 1.9 10 5 5 Day29 Day29 Day29 Day29 Day29 5mg/kg 5mg/kg 5rngkg 5mg/kg 5mgkg Avg SD Range Min Max N 04 1 202 3 10 042 205 30 1 043 196 3 06 044 202 295 045 203 288 202 300 3 9 196 288 205 310 5 5 5 2.3 13.0 2.2 11.4 2.2 11.7 2.1 11.0 2.0 10.6 2.2 12 0.1 1 2.0 11 2.3 13 5 5 Page 6 of 11 7 T-7098.1; ST35 Final Report Table 2: Table 2: Liver PFOS and M556 in the POSF dose Group Time Point Dose POSF animal # 9ROO-xxxLiver [PFOS] ppm Liver [M556]ppm Liver M556 PFOS eq (ppm) Day 1 5 m a g 03 1 2.35 0.125 0.11 Day 1 5 mg/kg 032 3.41 0.0957 0.09 Day 1 5 m a g 033 13.80 0.08 15 0.07 Day 1 5 mg/kg 034 3.73 0.1 14 0.10 Day 1 5 m a g 03 5 11.20 SE Avg 6.90 0.10 0.09 SD 5.22 0.02 0.02 Day 4 5 mg/kg 036 6.76 0.0166 0.01 Day 4 5 mg/kg 037 6.79 0.033 1 0.03 Day 4 5m a g 038 8.22 SE Day 4 5 mg/kg 039 12.50 0.0125 0.01 Day 4 5 mg/kg 040 19.70 SE Avg 10.79 0.02 0.02 SD 5.50 0.01 0.01 Day29 Day29 Day29 Day29 Day29 5mg/kg 5mg/kg 5mg/kg 5mg/kg 5mg/kg 04 1 2.55 <PQL 042 3.30 <PQL 043 7.54 <PQL 044 5.47 <PQL 045 2.57 <PQL Avg 4.29 SD 2.17 1. Liver M556 PFOS equivalents = Liver M556 concentration * (MW PFOSMW M556) = Liver M556 * ( 499/556) PQL = practical quantitation limit in liver for M556 = 0.060 ug/g SE = sample evaporated, no sample remaining Page 7 of 11 T-7098.1; ST35 Final Report Table 3: I I Table 3: Liver PFOS equivalents and Percent Dose in Liver Time Point IDose POSF Ihimal# PFOS eq in Total PFOS eq in PFOS eq. In % PFOS eq dosed Page 8 of 11 T-7098.1; ST35 Final Report lDay4 5 m g k g 03 7 0.094 7.0 0.7 968.2 Day4 5mgkg 03 8 0.0712 7.1 0.5 992.5 : b a y 4 5m*g 03 9 0.121 7.4 0.9 1009.4 lDay4 5mg/kg 040 0.119 7.2 0.9 989.6 I Avg 0.10 7.3 0.7 1005.2 I SD 0.02 0.3 0.2 Day29 5rngkg 04 1 0.0523 10.0 0.5 37.1 1004.0 lDay29 5mglkg 042 0.0928 9.7 0.9 1020.4 Day29 5mg/kg 043 0.2 1 9.9 2.11 975.6 Day29 5mgkg 044 0.152 9.5 1.4) 1002.5 ~Day29 5mgkg 045 0.0671 9.3 0.61 1009.4 Avg 0.11 9.7 1.11 1002.4 SD 0.07 0.3 0.61 16.5 1. There are 32.3 mL Plasma per Kg of rat. ~2T. otal PFOS equivalents in sera (ug) = PFOS equivalent concentration in sera @pm) * serum volume (ml) 3. PFOS equivalents in dose (ug) = Initial BW (g) * dose (mgkg) * MW PFOS/ MW M556 = BW*5*(499/556) 4. % PFOS equivalents dosed in serum = (PFOS equivalents in sera (ug)/ PFOS equivalents in dose (ug)) * 100 0.07 0.05 0.09 0.09 0.07 0.02 0.05 0.09 0.2 1 0.14 0.06 0.1 1 0.07 1 Page 9 of 1I DT 35; T-7098.1 Final Report Signatures: Prepared By: Advanced Research Toxicologist Study Toxicologist Andrew Seacat, Ph.D. Toxicology Specialist Study Director Date / Page 10 of 11 ! . DT 35;T-7098.1 Final Report References 1. 3M Medical Department, Study No. T-7098.1. 3M Strategic Toxicology Protocol Number: DT35. Title: PHARMACOKINETIC STUDY OF POSF IN RATS. 2. 3M Medical Department, Corporate Toxicology Strategic Toxicology Laboratory Standard Operating Procedure (TOX SOP) No. 0950 - GLP Program Procedure, 1999. 3. Hansen K. Laboratory Report Final Report of Data for POSF (T-7098.1) Phannacokinetic Study in Rats. Laboratory Report No. FACT-TOX-116. 3M Testing Laboratory 3M Environmental Technology & Safety Services 3M Environmental Laboratory Fluorine Analytical Chemistry Team (FACT) 2-3E-09 935 Bush Avenue, St. Paul, MN 55106. Page 11 of 11