Document 61pMLpvzBQn3DNjY0QvYdgGm

3M Specially Materials 3M Center St. Paul, MN 55144-1000 651 733 1110 BACK TO MAIN 3M VIA HAND DELIVERY January 18, 2001 Document Processing Center (740?) Office of Pollution Prevention and Toxics U.S. Environmental Protection Agency 401 M Street SW Washington, D.C. 20460 i. Attention: For Your Information Docket - Docket No. AR-226 Re: Information on Perfluorooctane Sulfonates and Related Compounds Dear Sir or Madam: This continues 3M 's voluntary submissions of data on perfluorooctane sulfonates and related compounds, as part of our ongoing dialog with EPA regarding fluorochemistry. ' Please find enclosed a supplemental submission containing a toxicokinetic study of perfluorooctane sulfonamide (FOSA). We just realized that this report had not yet been executed in final, and thus it was omitted from our recent December production. W e will provide an electronic copy of this material with a future submission. W e will continue to provide information on a regular basis as it becomes available; Very truly yours, Michael A. Santoro Director of Environmental, Health Safety and Regulatory Affairs Specialty Materials Markets January 18, 2001 Page 2 Enclosures - index and study cc: Dr. Charles Auer Dr. Oscar Hernandez BACK TO MAIN 3M . Building 236-1B-10 St. Paul, MN 55144-1000 651-733-6374 (phone) 651-733-1958 (fax) E-mail: masantoro@mmm.com ATTACHM ENT TO LETTER OF JANUARY 18, 2001 SUPPLEMENTAL SUBMISSION BACK TO MAIN TOXICOLOGY AND MEDICAL SURVEILLANCE FOSA Perfluoro o ctan esu lfo n am id e Pharmacokinetic 1. Final Report, Toxicokinetic Study o f Perfluorooctane Sulfonamide (PFOSA; T-7132.2) in Rats, 3M Strategic Toxicology Laboratory, 3M Reference Nos. T-7132.2; ST39, signed January 16,2001 (cover date August 11,2000). BACK TO MAIN FINAL REPORT Study Title: Toxicokinetic Study o f Perfluorooctane Sulfonamide (PFOSA; T-7I32.2) in Rats Authors: Andrew M. Seacat Ph.D. Toxicology Specialist 3M Medical Department, Corporate Toxicology Deanna J. Luebker M.S. Advanced Toxicologist 3M Medical Department, Corporate Toxicology Date: August 11,2000 Testing Facility 3M Strategic Toxicology Laboratory 3M Center 270-3S-05 Saint Paul, MN 55144 3M Strategic Toxicology ST39 T-7132.2 PFOSA PK study in rats BACK TO MAIN Summary: ` Purpose: The purpose o f this study was to assess the relative absorption, metabolism, and elimination kinetics of Perfluorooctane Sulfonamide (PFOSA) in the rat following a single oral dose. Methods: Two groups o f fifteen male rats each received a single dose of either 2% Tween 80, as the vehicle control, or 5 mg/kg PFOSA suspended in 2% Tween 80 by oral gavage. Perfluorooctanoate (POAA) was found at a level of 2.5 ng/ml in the dosing solution. The PFOSA was found to be 96 % pure Five rats from each dose group were sacrificed on days l, 4, and 29 post dose. Liver and sera were analyzed for PFOSA and its predicted metabolites using highpressure liquid chromatography/electrospray tandum mass spectrometry (HPLC/TrSMSMS). Results: The vehicle control group had no detectable levels of PFOSA or perfluorooctanesulfonate (PFOS) in liver or sera. The PFOSA dose group had decreasing levels of PFOSA, with evidence of conversion of PFOSA to PFOS in liver and sera throughout the study. Liver concentrations of PFOSA averaged 7.0 ppm, 3.7 ppm, and 0.1 ppm on days 1,4, and 29 post-dose, respectively, with an apparent liver half-life of elimination of 5.2 days. Serum concentrations of PFOSA averaged 0.3 ppm and 0.1 ppm on days 1 and 4 post dose respectively, and were undetectable by day 29 post-dose, with an apparent serum half-life of elimination estimated to be less than 4 cays. Liver PFOS concentrations were 28.2 ppm, 37.4 ppm and 23.8 ppm, representing 22.0%, 32.3%, and 25.2% of the PFOSA dosed, on days 1,4 and 29 post-dose respectively. Sera PFOS averaged 8.2 ppm, 8.6 ppm, and 4.6 ppm on days 1,4 and day 29 post dose respectively. The average total concentrations of PFOS equivalents (PFOS - PFOSA) in liver were 35.2, 41.1, and 23.9 ppm on days 1,4, and 29 post-dose, respectively. The average concentrations of PFOS equivalents in the sera increased from 8.5 ppm on day 1 post dose to 8.7 ppm on day 4 post dose and decreased to 4.6 ppm on day 29 post dose. The liver to sera PFOS concentration ratio increased throughout the study with values of 4.2. 4.8, and 5.4 or. days 1,4, and 29 post dose, respectively. Total PFOS equivalents in the liver and sera peaked on day 4 post dose at 35.4% and 6.0 % of the PFOS equivalents dosed, respectively. An average of 0.1 ppm POAA was found in the sera on day 1 and day 4 post dose, and equaled 31.2% and 24.1 %of the POAA residual present in the dose, respectively. Conclusions: Perfluorooctanesulfonamide was readily absorbed from the gut and found in the liver and sera following an oral exposure. PFOSA was progressively metabolized to PFOS, which accumulated in the liver as evidenced by the high concentration of PFOS detected in the liver, representing 22.0% 32.3% and 25.2% of the PFOSA dosed, on days 1,4 and 29 post dose respectively. The apparent half-live o f elimination o f PFOSA in the liver was approximately 5.2 days. The POAA present in the sera on days 1 and 4 post-dose could be accounted for by the residual POAA in the dosing solution, therefore, no direct evidence for metabolism o f PFOSA to POAA was found in this study. Page 2 o f 18 3M Strategic Toxicology ST39 T-7132.2 PFOSA PK study in rats BACK TO MAIN Introduction The objective of this study was to assess the potential for oral absorption, urinary and fecal clearance, and biological persistence of Perfluorooctanesulfonamide (PFOSA) in male Sprague Dawley rats after a single oral dose. Analysis of the serum and liver for PFOSA and potential metabolites of PFOSA was performed by LCMS. Urine and feces were also collected and may be analyzed by LCMS and perhaps other methods as deemed necessary. Previous studies on N-ethylperfluoroctanesuIfonamide (1), the N-ethyl derivative o f PFOSA, mistakenly concluded that PFOSA was the ultimate metabolite in rats (1,2); however, the gas chromatography analytical technique used in those studies was unable to detect PFOS. Dietary administration o f N-ethylperfluoroctanesulfonamidoethanol (N-EtFOSE) in rats, and in-vitro metabolism by hepatocytes results primarily in the formation o f PFOS with a minor amount of PFOSA (3,4, and 5). Furthermore, PFOSA has been identified as a potent direct uncoupler of mitochondrial respiration in-virro (6). Prior to the current study, direct metabolism of PFOSA to PFOS had not been demonstrated and the relative toxicokineiics o f PFOSA were unknown. Methods were recently validated for the quantitation,of PFOSA and its potential metabolite, PFOS, in serum and liver down to the low part per billion level (7). In-vivo studies were, thus, warranted to investigate the toxicokinetics of PFOSA. M aterials The test material was identified as Perfluorooctanesulfonamide (PFOSA); Lot number L-I0009. Initial analysis by GCMS determined that tne-staring material was over 99% pure (8). Qualitative and quantitative compositional results derived from :H and 19F-NMR analysis revealed that the isomer distribution was approximately 65.8% CF3 (CF2) X-S02-NH2 (Normal chain), 18.7% internal monomethyl branch and 11.2% Isopropyl branch (9). HPLC/MS characterization revealed low-level impurities of 9,600 ppm PFOS (0.96 %), and lower concentrations of several other amides (10). Perfluorooctanoate (POAA) was found in the dosing solution at a level of 2.5 fig/ml (0.25% of the nominal dose) (12). Based on the sum of the impurities, the purity of the PFOSA sample was determined to be approximately 96 % (8,9 & 10). Methods The protocol (11) and analytical methods (12) used in this study are briefly described below. Dose and Dosing Procedures: A single 5mg/kg dose of PFOSA was administered via oral gavage to 15 male Sprague Dawley rats (obtained from Harlan Laboratories) on day zero o f the study. The PFOSA was prepared as a 1% (1 mg/ml) uniform suspension in 2% Tween 80. A volume of 5 ml suspension / kg body weight was administered to each rat. The vehicle control group, consisting of 15 male Sprague Dawley rats (also obtained from Harlan Laboratories), received 2% Tween 80 in deionized water at a volume o f 5 ml/kg on day zero of the study. Page 3 of 38 3M Strategic Toxicology ST39 T-7132.2 PFOSA PK study in rats BACK TO MAIN Specimen Collection: Urine and feces collections were made on days 1 - 4 post dose from the animals designated for sacrifice on day 29 post dose group. Five animals from each dose group were euthanized by CO2 on days 1,4, and 29 post dose and gross necropsy was performed. Sera and liver were collected from each animal and flash-frozen in liquid nitrogen. Specimen Handling: Specimens (urine, feces, liver, and serum) were maintained at -70*0 and sent to Kris Hansen, Ph.D., 3M Environmental Technology and Safety Services, for analysis. Liver and sera samples were analyzed by 3M Environmental for the parent compound and metabolites. The limits of detection (LODs) for PFOSA were 3.5 ng/ml in the liver and 1.51 ng/ml in the sera. The LODs for PFOS were 8.45 ng/ml in liver and 1.74 ng/ml in the sera (12). Urine and feces were retained for possible future analysis as appropriate. Results , Control Groups: Bodv and Liver Weight The average initial body weight SD of the fifteen vehicle control male rats was 273.7 7.3 g. These rats lost an average of 2.2 g body weight from day zero to day 1 post dose . From day zero to day 4 post dose, the average body weight gain was 24.2 g and from day zero to day 29 post dose, body weight increased an average of 114.1 g . Liver weights and relative liver to body weight percentages are shown in Table 1A. Liver and Sera Concentrations Control animals had no detectable levels of PFOSA or PFOS in liver or sera at any time during this study (data not shown). . PFOSA Dose Groups: Bodv and Liver Weight The average initial body weight x SD of the fifteen PFOSA dosed male rats was 270.0 3.6 g. These rats lost an average of 4.5 g body weight from day zero to day 1 post dose . From day zero to day 4 post dose, the average body weight gain was 20.0 g and from day zero to day 29 post dose, body weight increased an average of 100.9 g . Liver weights and relative liver to body weight percentages were equivalent to control values (Table IB). Liver and Sera Concentrations PFOSA and PFOS were detected in the liver and sera of all animals dosed with PFOSA. Liver and sera levels of PFOSA were highest on day 1 post dose and dropped significantly between Page 4 of18 3M Strategic Toxicology ST39 T-7132.2 PFOSA PK study in rats BACK TO MAIN days 1 and 4 post dose and again between days 4 and 29 post dose (Tables 2 and 3). The average liver concentrations of PFOSA decreased from approximately 7.0 ppm ro 3.7 ppm to 0.1 ppm from days 1 to 4, to 29 post dose, respectively (Table 2). The half-life o f elimination o f PFOSA in the liver was calculated from a log linear regression of the liver PFOSA concentrations to be 5.2 days. The sera concentrations of PFOSA were 0.3 0.1 ppm, 0.1 0.0 and <LOD (1.74 ng/ml) on days 1,4, and 29 post dose, respectively (Table 3). The half-life of elimination of PFOSA in the serum cannot be calculated from this data, but is estimated to be less than 4 days. Average PFOS liver and sera levels peaked on day 4 post dose and declined by day 29 post dose to lower levels than found on day one. Average SD liver PFOS levels increased significantly from 28.2 6.1 ppm on day 1 post dose to 37.4 3.6 ppm on day 4 post dose. Levels then decreased significantly to 23.8 4.1 ppm by day 29 post dose (Table 2). Average sera PFOS values were approximately 8 ppm on day 1 and day 4 post dose, then decreased significantly to approximately 4.6 ppm on day 29 post dose (Table 3). Liver and Sera Concentrations as a Percentage of Dose The percentage of PFOSA dosed detected as PFOSA in the liver and sera peaked on day one post dose at 5.4% and 0.2 % respectively. By day 4 post dose, these levels had dropped to 3.2% and 0.0 % and by day 29 post dose the liver levels dropped to 0.1 % and the sera levels remained at 0.0% (Table 6). The percent PFOS equivalents dosed detected as PFOS in the liver and sera was 22.0% and 5.2 % on day 1 post dose and increased to 32.3% and 6.0% by day 4 post dose. On day 29 post dose, these levels had dropped to 25.2% and 4.1%, respectively (Table 6). Residual PFOS in the dose could account for 3.5%, 2.5%. and 5.6% of the PFOS found in sera, and 3.6%, 2.7%, and3.9 % of the PFOS found in liver on days 1,4, and 29 post dose, respectively (Tables 4 and 5). The percent PFOS equivalents dosed detected as PFOS equivalents in the liver and sera was 27.2 % and 5.3 % n day 1 post dose and increased to 35.4 % and 6.0 % by day 4 post dose. On day 29 post dose these levels decreased to 25.4 % and 4.1 % in the liver and sera respectively (Table 6). Sera PQAA Concentrations Perfluorooctanoate (POAA) was found at a level of 2.5 ng'ml in the dosing solution (12). Sera samples in the PFOSA dose group contained POAA at average values of 0.1 and 0.8 ppm POAA on days 1 and 4 post-dose respectively. These values equal 30.8% and 23.5%, respectively, of the POAA dosed (Table 7). Conclusions: No compound related effects were observed on body weigh:, liver weight, or liver/body weight ratios under treatment conditions. Both PFOSA and PFOS were found at high concentrations in the liver and sera of rats one day following an oral dose. Approximately 38% of the PFOSA dosed was converted to PFOS four Page 5 of 18 3M Strategie Toxicology ST39 T-7132.2 PFOSA PK study ir rats BACK TO MAIN days post-dose. PFOSA levels diminished in both sera and liver over time, with a liver half-life of approximately 5.2 days, and a much shorter residence period in the sera. Significantly more PFOS was found in the liver and sera at all time points than were present in the dose, therefore metabolic conversion of PFOSA to PFOS occurred. No direct evidence for metabolism of PFOSA to POAA was found in this study. The route of elimination o f PFOSA in this study was not determined. No glucuronide or other conjugates were analyzed for, and further investigation of possible conjugates in the urine and feces is warranted. It is possible that some o f the PFOSA was eliminated through the lung via expiration, but this has not been determined. The present results suggest that the PFOSA from the dose is progressively converted to PFOS and that the resulting PFOS is accumulated in the liver. Page 6 of 18 j M Strategic Toxicology ST39 T-7132.2 PFOSA PK study in rais Report Prepared by: Signatures Deanna Luebker, MS? Advanced Research Toxicologist ......... ft] , Andrew M. Seacat. Ph.D. Toxicology Specialist Report Reviewed by: / John L. Butenhoff, Ph.D., DABT, CITI Senior Laboratory Manager BACK TO MAIN .. - O l l i / M O I Date Olii 0! Date / Date o/ .... Page 7 of 18 3M Strategic Toxicology ST39 T-7132.2 PFOSA PK study in rats BACK TO MAIN References: _ 1. Grossman M.R. and Bowen J.M. (1990) Tissue analysis of iluorinated sulfonamide pesticide: an evaluation o f distribution, elimination, and potential for bioaccumulation in orally exposed rats. M.S. Thesis, Univ. of Georgia, Athens, GA. (also possibly published as: GrossmanMark R. and Bowen J.M. (1990) Tissue distribution and elimination of a iluorinated sulfonamide pesticide in rats. Fundam. Appl. Toxicol., but notfound). 2. Grossman M.R., Mispagel, M.E. and Bowen J.M. (1992) Distribution and tissue in rats during and after prolonged dietary exposure to a highly fluorinated sulfonamide pesticide. J Agric. Food Chem. 40. 2505 - 2509. .~ 3. Mulvana D.E. and Henion J. 1996. Qualitative investigation of the in-vitro metabolism of T6292, T-6293, T-6294 and T-6295 by rat and human hepatocytes using ion spray LC/MS and LC/MS/MS. Analytical Report: 96ADEM01.3M, Advanced Bioanalytical Services, Inc. 45pp. 4. Poon G.K., Lowes S. 1998. Additional Characterization of metabolites of T-6292, T-6293 and T-6294 from rat and human hepatocytes by TurboIonSpray LC/MS and LC/MS/MS. Semi-quantitative analysis of T-6295 ir. rat and human hepatocytes incubated with T-6292, T-6293 and T-6294 by LC/MS/MS. Analytical Report: 96AGKP01.3M, Advanced Bioanalytical Services, Inc. 69pp. 5. Seacat A.M., Thomford P.J.. Hansen K.J. and Butenhoff J.L. Sub-Chronic Dietary N-Ethyl Perfluorooctanesulfonamido Ethanol Toxicity in Ra'ts. (in preparation, 8/11/2000), 6. Wallace K.B. and Starkov A. 1998. The effect of perfluorinated arylalkylsulfonamides on bioenergetics of rat liver mitochondria. Dept, of Biochemistry and Molecular Biology, University of MN School of Medicine. Duluth, MN 55812, LrSA. Supported by a grant from 3M Company 7. K.J. Hansen, L.A. Clemen, M.E. Ellefson, H.O. Johnson. (1999). Compound Specific Characterization of Organic Fluorochemicals in General Population Human Sera Samples. 3M Environmental Lab, St. Paul, MN 55133. 8. Payfer R.M. GC/MS analysis of PFOSA (L-10009). SAScC Analytical Request No. 59426. Report 9/24/99. 3M-SA&C Lab Building 236-2B-11. ' 9. Tom Kestner. Chemical Characterization of PFOSA, L-10009, by 1H and 19F-NMR Spectroscopy Requests # 59426.3M Specially Adhesives & Chemicals Analytical Laboratory / SMMD-236-2B-11, September 25, 1999. 10. DeRoos F.L. Characterization of PFOSA Samples, T-7132-1 (L-10009) and TN-A-1584. Request # A-151254. Report 10/7/99. Corporate Analytical Technology Center, Building 201-1-29, CATC - Chromatography Group. 'Page 8 of 18 3M Strategic Toxicology ST39 T-7132.2 PFOSA PK study in rats BACK TO MAIN 11. Seacat & Luebker. Protocol for Study No. T -7!32.2; ST-39, PHARMACOKINETIC STUDY OF PFOSA IN RATS. 3M Medical Department, Corporate Toxicology. October 1999. 12. Laboratory Report: Analytical Report ofData for PFOSA (T-7132.2) Pharmacokinetic Study in Rats (Sera and Liver). Laboratory Report No. FACT-TOX-145 (W2814). Testing Laboratory 3M Environmental Technology & Safety Services, 3M Environmental ' Laboratory, Fluorine Analytical Chemistry Team. Laboratory Contact: Kris Hansen, Ph.D. 13. Alman and Dittmer. Blood and Other Body Fluids. FASEB, 1971, p5 Page 9 o f 18 3M Strategic Toxicology ST39 T-7132.2 PFOSA PK study in rats List of Tables: Table 1: Biological Parameters 1A: Control Group IB: PFOSA Dose Group. Table 2: Liver PFOSA and metabolites in the PFOSA Group. Table 3: Serum PFOSA and metabolites in the PFOSA Group Table 4: Percent Dose; Liver PFOS equivalents in PFOSA Group. Table 5: Percent Dose: Serum PFOS equivalents in PFOSA Group. Table 6: Summary: Percent of dose in sera and liver. Table 7: Serum POAA in the PFOSA Group. , BACK TO MAIN Page 10 o f 18 3M Strategic Toxicology ST39 T-7132.2 PFOSA PK. study in rats BACK TO MAIN Table 1A: Biological Parameters. Control Group T im e Day 1 Day 1 Day 1 Day 1 Day 1 Dose Control Control Control Control Control Avg anim al # Initial Term inal BW Gain 9R03-xxx BW (g) BW (g) (9) 230 282.5 282.0 -0.5 231 272.1 268.4 -3.7 2321 277.4 277.1 -0.3 233 274.1 274.8 0.7 234 267.6 i 260.5 -7.1 274.7 272.6 -2.2 Liver Liver w t as w t(g) % of BW 12.0; 4.3% 11.2 12.4 4.2% 4.5% 11.51 4.2% 11.2 4.3% 11.7 4.3% SD 5.6 8.3 3.2 0.5 0.1% Day 4 Control 235 266.9 294.5 27.6 11.6 3.9% Day 4 Control Day 4 Control i 236 275.8 237 261.9 301.8 283.4 26.0 21.5 11.4 11.9 3.8% 4.2% Day 4 Control Day 4 Control 238 279.0 239 268.3 302.7 291.2 23.7 22.9 11.3 10.7 3.7% 3.7% Avg 270.4 294.7 24.3 11.4 3.9% SD 6.9 8.0 2.4| 0.4 0.2% Day 29 Control Day 29 Control Day 29 Control Day 29 Control Day 29 Control Avg 240 273.5 241 270.5 242 280.2 243 289.9 244 266.3 276.1 371.8 380.5 389.4I 444.0I 365.2 390.2! 98.3I 110.01 109.2 154.1 98.9 114.1 13.9 14.4 14.2 17.5 14.3 14.9 3.7% 3.8% 3.6% 3.9% 3.9% 3.8% SD 9.2 31.4 23.0 1.5 0.1% Page 11 of 18 3M Strategie Toxicology ST39 T-7132.2 PFOSA PK study in rats BACK TO MAIN Table 1B; Biological Parameters. PFOSA Dose Group Time ! Dose anim al # Initial Term inal BW Gain Liver Liver w t as Day 1 Day 1 | 9R03-xxx BW (9) ;5 mg/kg 245 263.2 15 mg/kg 246 266.1 BW (g) 257.8 259.2 .(9) _ w t (g) % o f BW -5.4 10.41 4.0% -6.9 10.2 3.9% Day 1 5 mg/kg i 247 266.5 264.1 -2.4 10.9 4.1% Day 1 |5 mg/kg Day 1 )5 mg/kg 248 267.2 249 268.7 264.0 264.3 -3.2 10.1 -4.4 10.7 3.8% 4.0% lAvu 266.3 261.9 -4.5 10.5 4.0% ISD 2.0 3.1 1.8 0.3 0.1% Day 4 |5 mg/kg 250 266.6 286.3 19.7 11.3 3.9% Day 4 5 mg/kg 251 272.1 294.6 22.5 11.7 4.0% Day 4 5 mg/kg 252 271.7 294.5 22.8 12.1 . 4.1% Day 4 |5 mg/kg 253 271.8 291.4 19.6 11.3 3.9% Day 4 |5 mg/kg 254 272.9 288.4 15.5 11.7 4.1% . |Avg 271.0 291.0 20.0 11.6 4.0% |SD 2.5 3.7 2.9 0.3 0.1% Day 29 I5 mg/kg 255 275.2 385.4 110.2 16.7 4.3% Day 29 5 mg/kg 256 273.6 380.3 106.7 14.4 3.0% Day 29 5 mg/kg 257 271.4 373.9 102.5 13.9 3.7% Day 29 5 mg/kg 258 274.5 371.4 96.9, 14.6 3.9% Day 29 5 mg/kg 259 268.7 356.7 88.0 12.7 3.6% Avg 272.7 373.5 100.9 j 14.5 3.9% SD 2.6 10.9 _____ *LZ 1.5 0.3% Page 12 o f) 8 3M Strategic Toxicology ST39 T-7132.2 PFOSA PK study in rats BACK TO MAIN Table 2: Liver PFOSA and metabolites in the PFOSA dose Group animal tt 9 R 0 3 - Liver (PFOS) Liver [PFOSA] Liver PFOSA PFOS Time Dose XXX ppm ppm eq p p m 1 Day 1 5 mg/kg 245 23.7 6.02 6.02 Day 1 5 mg/kg 246 32.3 7.02 7.02 Day 1 5 mg/kg 247 36.2 6.24 6.24 Day 1 5 mg/kg 248 21.2 6.66 6.66 Day 1 5 mg/kg 249 27.8 8.81 8.81 Avg 28.2 6.95 6.95 SD 6.1 1.11 1.11 Range Min 21.2 6.02 6.02 Max 36.2 8.81 8.8! n 55 5 5 Day 4 5 mg/kg 250 36.8 3.52 3.52 Day 4 5 mg/kg 251 32.6 3.47 5.47 Day 4 5 mg/kg 252 42.6 3.96 . 3.96 Day 4 5 mg/kg 253 36.9 3.45 3.45 Day 4 5 mg/kg Avg 254 38.3 37.4 * 4.11 3.70 w 4.11 3.70 SD 3.6 0.31 Range Min 32.6 3.45 0.31 3.45 Max 42.6 4.11 4.11 n 55 5 5 Day 29 5 mg/kg 255 20.3 .... 0.09 0.09 Day 29 5 mg/kg Day 29 5 mg/kg 256 20.6 257 22.0 0.02 0.10 0.02 0.10 Day 29 5 mg/kg . 258 26.2 0.14 0.14 Day 29 5 mg/kg Avg 259 29.9 2 3 .8 ** 0.30 0 .1 3 * f ** 0.30 0.13 SD Range Min . 4.1 20.3 0.11 0.02 0.11 0.02 Max 29.9 0.30 n 55 5 0.30 5 Assum ing a 1:1 ratio of PFOSA:PFOS equivalents; Liver PFOSA PFOS equivalents =Liver PFOSA * = significantly different from day 1 values by a two tailed T-test (p<0.05) ' = significantly different from day 4 values by a two tailed T-test (p<0.05) PQL - practical quantitation limit in liver (PFOS = 30 ng/g; PFOSA 10ng) MDL = method detection limit in liver: (PFOS = 15 ng/g, PFOSA = 5 ng/g) Page 13 of 18 j M Strategic Toxicology ST39 T-7132.2 PFOSA PK study in rats Table 3: Serum PFOSA and m eta b o lite s in the PFOSA Group . BACK TO MAIN Time 5ay l Day 1 Day l Day 1 Dose 5 mg/kg 5 mg/kg 5 mg/kg 5 mg/kg an im al n 9 R 0 3 - Sera (PFOSJ XXX ppm m 6.5 246 10.8 247 5.4 248 6.7 Sera [PFOSA] ppm 0.26 0.44 0.26 0.33 Sera PFOSA PFOS eq ppm1 or 0.44 0.26 0.33 Day 1 5 mg/kg Avg SD Range Day 4 n. 5 mg/kg 249 Min Max 5 250 7.3 8.2 .1-8 6.7 10.8 5 7.8 0.28 0.31 0.08 0.26 0.44 5 0.06 0.28 0.31 0.08 0.26 0.44 5 0.06 Day 4 5 me/kg 251 7.5 0.06 0.06 Day 4 5 mg/kg 252 11.3 0.08 0.08 Day 4 5 mg/kg 253 8.6 0.06 0.06 Day 4 Day 29 Day 29 Day 29 Day 29 5 mg/kg Avg SD Range n 5 mg/kg 5 mg/kg 5 mg/kg 5 mg/kg 254 Min Max 5 255 256 257 258 7.9 8.6 1.6 7.5 n .a 5 3.6 4.0 4.0 4.1 0.06 0.06 9 0.01 0.06 U.Uts 5 <LOD <LOD <LOD 10D 0.06 0.06 0.01 0.06 CTF 5 Day 29 5 mg/kg Avg SD lange n 259 Viin Max 7.4 4.6 1.5 3.59 7.36 5 * j ** 0.0 0.0 5 5 1. Assuming a 1:1 ratio of PFOSA :PFOS equivalents, Sera PFOSA PFOS equivalents =Sera PFOSA concentration = significantly different from day 1 values by a two tailed T-test (p<0.05) * ' = significantly different from day 4 values by a two tailed T-test (p<0.05) M D l = method detection limit (PFOS = 4.75 ng/ml; PFOSA = 2.89 ng/ml) LOD = limit of detection (PFOS = 1.74 nq/ml: PFOSA = 151 na/mO Page 14 o f 18 BACK TO MAIN TM Strategic Toxicology S I 39 T-7132.2 l'I'OSA I'K simly in rats Table 4: P ercent D ose Liver PFOS equivalents in PFOSA G roup Time Dose Day 1 Day 1 Day 1 Day 1 Day 1 5 mg/kg 5 mg/kg 5 mg/kg 5 mg/kg 5 mg/kg Avg SD Range Day 4 Day 4 Day 4 Day 4 Day 4 n 5 rnglkg 5 mg/kg 5 mg/kg 5 mg/kg 5 mg/kg Avg SD Range Day 29 Day 29 Day 29 Day 29 Day 29 n 5 mg/kg S mg/kg 5 mg/kg 5 mg/kg 5 mg/kg Avg SD Range n animat # 9R03-xxx 245 246 PFOS eq concentration in liver (ppm )1 29.71 39.3 1Total PFOS eq in w hole liver (ug)* 309.1 401.1 PFOS eq in % PFOS eq d o se (u g )1 d o s e d found ir liver * 1316.0 23.5 1330.5 30.1 Residuai Vof PFOS in liver PFOS in do se rom residual a i ! PFOS in______________________ d o s e (%)* 12.6 4.09% 12.8 3.18% 247 42.4 462.6 1332.5 34.7 12.8 2.77% 248 27.9 281.4 1336.0 21 1 12.8 4.56% 249 36.6 391.7 1343.5 29.2 12.9 3.29% \ 35.2 369.2 1331.7 27.7 12.8 3.58% 6.2 73.4 10.1 5.5 0.1 0.73% Min 27.9 281.4 1316.0 21.1 12.6 2.77% Max 42.4 462.6 1343.5 34.7 12.9 4.56% 55 5 55 5 5 250 40.3 455.6 1333.0 34.2 12.8 2.81% 251 36.1 422.0 1360.5 31.0 13 1 3.09% 252 46.6 563.4 1358.5 41.5 13.0 7.31% 253 40.4 456 0 1359.0 33.6 13.0 2.86% 254 42.4 406.2 1364.5 36.4 13.1 41.1 478.6 * 1355.1 35.3 * 13.0 2.64% 2.74% 3.8 54.2 12.6 3.9 0.1 0.29% Min 36.1 422.0 1333.0 31.0 12.8 2.31% Max 46.6 563.4 1364.5 41.5 13.1 3.09% 5 5.0 5.0 5.0 5.0 5 5 255 20.4 340.4 1376.C 24.7 13.2 3.80% 256 20.6 296.9 1368.0 21.7 13. 4 42% 257 22.1 307.2 1357.C 22.6 13.0 4.24% 258 26.3 384.6 1372.E 280 13.2 3.43% 259 30.2 383.6 1343. 28.6 12 9 23.9 a * 342.5 * 13 63.4 25.1 A* 13.1 4.2 41.2 13.2 3.1 o.i 3.36% 3.87% 0.47% Min 20 4 296.G 1343. 21.7 12.9 3.36% Max 30.2 384.6 1376() 28.r 13.2 4.42% rC i r 5 7 PFOS equivalents mtWcr (ppm) = Liver PFOS (ppm) + Liver PFOSA PFOS equivalents (ppm); 2. lolal PFOS equivalents in whole liver (ug) = PFOS equivalent concentration in liver (ppm) * liver weight (g); 3. Assuming a \ \ ratio of PFOSA.PFOS equivalents. PFOS equivalents in dose (ug) = Initial BW (g) * dose (mg/kg) = BW*5; 4. % PFOS eq dosed as PFOSA in liver =(PFOS equivalents in whole liver (ug)/ PFOS equivalents in dosejMOO; 5. Restduat PFOS in dose (ug) * dose'O.96% = (5mg/kg*BW)0.0096; 6. % of I'FOS in tiver ironr residual I'FOS in dose = (residual I'FOS in dose / lolal PFOS eg in iivei+V I4)*100; * Significantly different from day 1 values by a two failed T-lest (p<0.05); Significantly different from day 4 values by a two tailed T-tesI (p<0.05); PQL = practical quantitation limit (PFOS = 30 ng/g; PFOSA 1 0 n g ) ________________________________ ______ __________ .-- ----- Page 15 oF I ft BACK TO MAIN 3M Strategic Toxicology ffl'39 T-7132.?. I'FOSA PK. study in rats Table 5: P ercen t D ose: Serum PFOS equivalents in PFOSA Group Time Day 1 Day 1 Dose animal PFOS eq cone PFOS c q Liver/serum Total PFOS It 9RD3- in Serum co n e in PFOS ratio Eq in whole XXX (ppm) ' liver (ppm) (ppm/ppm) fiver (ug) S mg/kg 5 mg/kg 24b 246 72 11.2 29.7 39.3 4.1 309.3 3.5 401.4 Serum Total PFOS Liver/ Scrum PFOS eq % PFOS eq R esidual % of PFOS in volume Eq in Serum (ml)* (u g )1 PFOS Eq ratio4 in dose dosed Found PFOS in seru m from (u g )4 in serum* d o se residual PFOS (ug)T I n d o s e (%)* 8.3 60.0 5.2 1316.0 4.6 12.C 21.1% 8.4 94.1 4.3 1330.5 7.1 12.8 13.6% Day 1 5 mg/kg 247 96 42.5 4.4 462.9 8.5 81.9 5.6 1332.5 6.1 12.8 " ' 15.6% Day 1 5 mg/kg 248 7 0 2 7 9 4.0 281.7 8.5 59.6 4.7 1336.0 4.5 12.8 21.5% Day 1 5 mg/kg 249 7.6 36.6 4.8 392 1 8.5 65.0 6.0 1343.5 4.8 12 9 19.8% Avg B.5 35.2 4.2 369.5 8.5 72.1 5.2 1331.7 5.4 12.8 17.7% SO 1.8 6 2 0.5 73.4 0.1 15.3 0.7 10.1 1.1 0.1 3.5% Range Min 7.0 27.9 3 5 281.7 8.3 59.6 4.3 1316.0 4.5 12.6 13.6% Max 11.2 N 55 42.5 5.00 4.8 462.9 8.5 5 55 94.1 5 6.0 1343.5 55 7.1 12.9 55 21.5% 5 ay 4 ay 4 Day 4 Day 4 Day 4 5 mg/kg 250 5 mg/kg 251 5 mg/kg 252 5 mg/kg 253 5 mg/kg 254 Avg SD Range Min 7.8 7.5 11.4 8./ no 8.7 1.6 7.5 40.3 36.1 40.6 40.4 42 4 41.2 3.8 36.1 5.1 455.8 9.2 72.5 4.8 422.2 9.5 71.7 4.1 563.6 9.5 10B.2 4.7 456.1 9.4 81.6 5.3 496.4 9.3 74.2 4.8 47B.8 9.4 81.7 0.5 54.2 0.1 15.3 4.1 422.2 9.2 71/ 6.3 1333.0 5.9 1360.5 5.2 1358.5 5.6 1359.0 6 ./ 1364.5 5.9 1355.1 0.6 12.6 5.2 1333.0 5.4 12.8 17.6% 5.3 13.1 18.2% 8.0 13. 12.1% 6.0 13.0 16.0% 5.4 13.1 17.6% 6.0 13 0 " 15.9% 1.1 0.1 2.5% 5.3 12.8 12.1% Max 11.4 46 .F) 5.3 5G3.6 9.5 1QB.2 6.7 1364.5 8.0 13.1 18.2% N 55 5 5 55 5 55 55 5 Day 29 5 mg/kg 255 3.6 20.4 5.7 340.4 12.4 44.7 7.6 1376.0 3.2 13.2 29.6% ay 29 5 mg/kg 256 4.0 20.G 5.7 296.9 12.3 48.9 6.1 1368.0 3.6 13.1 26.9% Day 29 5 mg/kg 257 4.0 22 1 5.5 307.3 12.1 46.8 6.3 1357.0 3.6 13 0 26.7% Day 29 5 mg/kg 258 4.1 26.3 6.4 364.6 12.0 49.5 7.8 1372.5 3.6 13.2 ........ 26.6% Day 29 5 mg/kg Avg 259 7 4.6 tt 30.2 23.9 * 4.1 303.6 11.5 5.4 342.6 12.1 84.8 55.3 4.5 1343.5 6.5 1363.4 6.3 12.9 4.1 13.1 ' 15.2% 23.7% SD Range Min Max 1.5 4.2 3.6 20.4 7.4 30.2 0.8 41.2 0.4 4.1 296.9 11.5 6.4 384.6 12.4 16.6 44.7 84.8 1.3 13.2 4.5 1343.5 7.8 1376.0 1.3 0.1 3 2 12.9 6.3 13.2 5.6% 15.2% 29.6% N 55 5 55 5 5| 5 55 5 1. PFOS equivalents in sera (ppm) = sera PFOS (ppm) + sera PFOSA PFOS equivalente Ippm); 2 1lieie are 32.3 mL Plasma pet Kg o( rat. (9); 3 Tolsi PFOS equivalents in sera (uij) = PFOS equivalent concentration in sera (ppm) * serum volume (ml); 4. Liver/ Serum total PFOS Eq ratio = total PFOS equivalents in wfiole liver (ug)/ total PFOS equivalents in serum (ug); 5. Assuming a 1:1 ratio ol PFOSA.PFOSA PFOS equivalents, PFOS equivalents in dose (ug) = Initial BW (g) *dose (mg/kg) = BW*5; 6.% PFOS equivalents dosed In serum - (PFOS equivalents in sera (ug)/ PFOS equivalents in dose (ug))MOO; 5. Residual PFOS in dose (ug) - dose`O.96% = (5mglkg'BW)0 0096; 6. "/.o l f'FOS in sera from residui! I'FOS m dole *=(residuai PFOS in dose/ miai I'FOS cq in icr)M00;** Sign, ditferent from dy4 values by a two tailed T-test (p<0.05); MDL - method detection limit in sera: (PFOS = 4.75 ng/ml; PFOSA 2.89 ng/ml) ______________ l'nnc IAt\i IV 3M Strategic Toxicology SI 39 T-7132.2 PI-'OSA PK. study in rats BACK TO MAIN Table 6 - Summary: % + SD of Dose or of PFOS Equivalents Dosed Detected in Liver & Sera _________________________________________ Day 1 post dose day 4 p o s t dose day 29 p o st d o se % P F O S A d o s e d d etected as P F O S A in: liver seta fiver & sera 5.4 i 0.9 0 .2 0.1 5.6 0.9 3.2 0 .3 0.0 0.0 3.2 0 .3 % P F O S E a u iv ale n ts d o se d d etected as PFOS in: liver sera liver & sera j 22.0 + 5.2 5.2 1 1 27.1 6 .2 32.3 3.7 6.0 .1 1 38.3 4.8 NA = not analyzed PFO S equivalents (ppm ) - PFOS (ppm ) 4 PFOSA PFOS equivalents (ppm) 0.1 .0.1 0 0 0.0 0.1 0.1 25.2 + .3 0 4.1 1 .2 29.3 3 .8 . _. 3M Strategic Toxicology ST39 T-7132.2 I'FOSA PK study in rats Table 7: Serum POAA in th e PFOSA G roup. i oi at 7o 1UAH POAA in dosed animal H Sera lolul POAA Tound us 9R03- POAA Serum in dose POAA in Ti m* Dose X X X (ppm) (ug) (ug) sera Day 1 Day 1 5 mg/kg 5 mg/kg 245 0.11 0.90 3.29 27.3% ' 246 0.15 1.23 3.33 37.0% Day 1 5 mg/kg 247 0.13 i .09 3.33 32.8% Day 1 5 mg/kg 248 0.1 0.88 3.34 26.3% Day 1 5 ing/kg Avg SD Range 249 Miu 0.12 0.12 0.02 0.10 1.02 1.02 0.15 0.88 3.36 3.33 0.03 3.29 30.5% 30.8% 4.3% 26.3% Max 0.15 1.23 3.36 37.0% n 5 5 5 55 Day 4 5 mg/kg 250 0.07 0.63 3.33 18.9% Day 4 5 mg/kg 251 0.07 0.64 3.40 18.9% Da y 4 5 mg/kg 252 0.11 1.00 3.40 29.4% Da y 4 5 mg/kg 253 0.10 0.95 3.40 28.0% Day 4 5 tng/kg 254 0.08 0.76 3.41 22.2% Avg 0.08 0.80 3.39 2 3 .5 % SD 0.02 0.17 0.03 5.0% Range Min 0.07 0.63 3.33 18.9% Max 0.1 1 1.00 3.41 29.4% n 5 55 5 5 Lirnil of detection (LOD): POAA 24.7 ng/ml NA. Not analyzed * based on work by Kris Hansen finding 2.5 ug/rn) POAA in dosing solution 1There are 32.3 mL Plasma per Kg of rat.(13) BACK TO MAIN f'agc IH oi IH