Document Eq5Xk7KnG9e04rLajgZRDq5e4

3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 Table 7: Ambient-Temperature Stability of PFOS in Rat Serum After 24 Hours Run 11 QC (Theoretical Cone.) QC 1 (0.2 pg/mL) Mean RSD (%) %DEV from 0 Hour OHour 0.194 0.197 0.184 0.192 0.184 0.190 3.03 NA PFOS (pg/mL) 2.5 Hour 6 Hour 0.187 0.177 0.181 0.179 0.184 0.180 0.180 0.183 0.178 ' 0.189 0.182 0.182 1.91 2.52 -4.20 -4.45 24 Hour 0.190 0.187 0.191 0.182 0.190 0.188 2.01 -1.16 6.17 6.17 6.36 6.55 QC2 (6 pg/mL) 6.13 6.30 6.38 6.41 6.26 6.35 6.15 6.28 6.22 6.24 6.24 6.11 6.53 6.85 6.66 6.81 Mean RSD (%) %DEV from 0 Hour 6.22 6.25 6.28 1.82 0.797 1.86 NA 0.525 1.05 6.68 2.16 7.42 18.2 18.0 18.1 19.1 QC 3 (18 pg/mL) 17.1 18.0 18.0 19.4 17.6 17.7 17.6 19.2 .17.7 17.9 18.2 19.4 17.0 18.0 18.0 19.7 Mean RSD (%) %DEV from 0 Hour 17.5 17.9 18.0 2.84 0.719 1.26 NA 2.33 2.39 19.4 1.19 10.5 NA: Not Applicable RSD = (SD/Mean) x 100 %DEV from 0 Hour = [(X Hour Cone. - 0 Hour Conc.)/0 Hour Cone.] x 100 3M Environmental Laboratory ADVANCED B IO AN A LYTIC A L S E R V IC E S , INC. Page 26 of 50 99VDJA01 .MI.DOC Page 197 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Table 8: Freezer Stability of PFOS in Flat Serum at -20 C QC Nominal Concentration QC 1 (0.2 ug/mL) 0 Hour (Run 2)* 0.213 0.209 0.208 0.208 0.207 PFOS (tig/mL) 7 Days 13 Days (Run 6)* (Run 3)** 0.2215 0.211 0 .22 :2 0.210 0.218 0.217 0.219 0.214 Mean RSD (%) %DEV from 0 Hour 0.209 1.24 NA 0.221 1.32 5.72 0.213 1.55 1.84 6.02 6.22 5.97 QC2 5.71 6.15 5.84 (6 ug/mL) 6.00 6.39 6.07 6.06 6.27 5.99 5.69 Mean 5.89 6.26 5.97 RSD (%) 3.08 1.62 1.58 %DEV from 0 Hour NA 6.19 1.20 20.2 19.1 18.8 QC 3 18.6 19.5 18.2 (18 ug/mL) 18.9 19.8 18.8 18.2 19.3 17.9 18.9 Mean RSD (%) %DEV from 0 Hour 18.9 3.96 NA 19.4 18.4 1.37 2.42 2.52 -2.76 NA: Not Applicable RSD = (SD/Mean) x 100 %DEV from 0 Hour = [(Cone. - 0 Hour Conc.)/0 Hour Cone.] x 100 From Protocol FACT-TOX-110 From Protocol FACT-TOX-111 33 Days (Run 16) 0.245 0.234 0.228 0.232 0.235 3.17 12.3 6.45 6.13 6.58 6.37 6.38 2.97 8.28 18.6 18.8 18.7 18.7 18.7 0.395 -1.30 3M Environmental Laboratory ADVANCED B IO AN A LYTIC A L S E R V IC E S , INC. Page 27 of 50 99VDJA01.MI.DOC Page 198 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Table 9: Stability of PFOS in Rat Serum After Three Freeze/Thaw Cycles Run 10 PFOS (pg/mL) QC (Theoretical Cone.) QC 1 (0.2 pg/mL) 0 Cycle 0.187 0.186 0.182 0.189 3 Cycles 0.177 0.177 0.185 0.185 0.182 0.189 Mean RSD (%) %DEV from 0 Cycle 0.185 1.70 NA 0.182 3.12 -1.38 6.U 6.59 QC2 (6 pg/mL) Mean RSD (%) %DEV from 0 Cycle 6.44 6.32 6.19 6.31 6.34 1.66 NA 6.61 6.44 6.39 6.40 6.49 1.63 2.35 QC 3 (18 pg/mL) 16.9 19.2 17.3 19.1 17.5 19.2 18.0 19.6 17.4 18.7 Mean RSD (%) %DEV from 0 Cycle 17.4 2.31 NA 19.2 1.80 10.1 NA: Not Applicable RSD = (SD/Mean) x 100 %DEV from 0 Cycle = [(3 Cycle - 0 Cycle)/0 Cycle] x 100 3M Environmental Laboratory ADVANCED B IO AN A LYTIC A L S E R V IC E S , INC. Page 28 of 50 99VDJA01.MI.DOC Page 199 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Table 10: Reproducibility of Reinjecting Extracted Samples Containing PFOS after 27 Hours in Reconstitution Solution QC1 (0.2 pg/mL) Mean RSD (%) RE (%) QC2 (6 pg/mL) Mean RSD (%) RE (%) QC3 (18 pg/mL) Mean RSD (%) RE (%) NA: Not Applicable RSD = (SD/Mean) x 100 RE = [(Mean/Theoretical)-1] x 100 PFOS (ug/mL) t=0 Hours t=27 Hours (Run 10) (Run 12) 0.187 0.187 0.186 0.182 0.182 0.188 0.189 0.188 0.182 0.190 0.185 0.187 1.70 1.61 -7.50 -6.33 6.44 6.39 6.44 6.02 6.32 6.19 6.19 6.55 6.31 6.22 6.34 6.28 1.66 3.23 5.62 4.59 16.9 17.3 17.5 18.0 17.4 17.4 2.31 -3.29 17.3 18.2 18.1 17.3 17.6 17.7 2.40 -1.73 3M Environmental Laboratory ADVANCED B IO AN A LYTIC A L S E R V IC E S . IN C . Page 29 o f 50 99VDJA01.MI.DOC Page 200 3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 Table 11: Extraction Recovery of PFOS from Rat Serum Theoretical Cone. Pre-Extraction Post-Extraction in serum (pg/mL) Response (Area Ratio) Response (Area Ratio) % Recovery* PFOS 0.03160 0.02880 110 Spike 1 0.02850 0.03010 94.7 (0.25 pg/mL) 0.02930 0.02950 99.3 Run 14 0.02790 0.02930 95.2 0.03050 0.02960 103 Mean 0.02956 0.02946 100 PFOS Spike 2 (4 pg/mL) Run 14 0.4597 0.4751 0.4776 0.4724 0.4772 . 0.5270 0.5304 0.5335 0.5298 0.5392 87.2 89.6 89.5 89.2 88.5 Mean 0.4724 0.5320 88.8 PFOS Spike 3 (16 pg/mL) Run 14 1.649 1.679 1.719 1.535 1.612 1.874 1.857 1.869 1.887 1.864 88.0 90.4 92.0 81.4 86.5 Mean 1.639 1.870 87.6 Tetra-H-PFOS Spike IS (4 pg/mL) Run 13 Mean 0.4068 0.3884 0.4043 0.3921 0.4078 0.3999 0.4460 0.4423 0.4445 0.4512 0.4412 0.4450 91.2 87.8 91.0 86.9 92.4 89.9 (Individual Response for Pre-Ext./Indivic ual Response for Post-Ext.) x 100 3IVI Environmental Laboratory ADVANCED B IO AN A LYTIC A L S E R V IC E S , INC. Page 30 of 50 99VDJA01.MI.DOC Page 201 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 8. FIGURES Figure 1: Full-Scan Single MS Mass Spectrum of PFOS Spectrum from PFOS Q1 ' 3.19e7cps 3IVI Environmental Laboratory ADVANCED B IO AN A LYTIC A L S E R V IC E S , INC. Page 31 of 50 99VDJA01 .MI.DOC Page 202 3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 Figure 2: Full-Scan Single MS Mass Spectrum of Tetra-H-PFOS Spectrum from Tetra-H-PFOS Q1 ' .94e7 cps 3M Environmental Laboratory ADVANCED B ID AN A LYTIC A L S E P V IC E S , IN C . Page 32 of 50 99VDJA01.MI.DOC Page 203 3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 Figure 3: Mass Chromatograms of PFOS and its Internal Standard in Control Blank Rat Serum Extract m/z = 427.0 1.07e3 cps Tims, min All quantitation results and figures were prepared from non-smoothed data. Analvte PFOS Tetra-H-PFOS (IS) Ion m/z = 499.0 m/z = 427.0 Retention Time 3.1 2.9 3M Environmental Laboratory ADVANCED B IO AN A LYTIC A L S E R V IC E S , IN C. Page 33 of 50 99VDJA01.MI.DOC Page 204 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Figure 4: Mass Chromatograms of PFOS in a Rat Serum Extract Sample Containing Internal Standard Only (Zero Sample) m/z = 427.0 2.34e5 cps 90 80 70 60 50 40 30 20 - 10 3 m/z =499.0 > Js Q Time:, min 3.78e2 cps Time, min All quantitation results and figures were prepared from non-smootbed data. Analvte PFOS Tetra-H-PFOS (IS) lea m/z = 499.0 m/z = 427.0 3.1 2.9 3M Environmental Laboratory ADVANCED BIO AN A LYTIC A L S E R V IC E S . IN C . Page 34 o f 50 99VDJA01 .MI.DOC Page 205 3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 Figure 5: Mass Chromatograms of Calibration Standard 1 in Rat Serum Extract Containing PFOS (0.0:5 pg/mL) and the Internal Standard m/z = 427.0 3.82e5 cps All quantitation results and figures were prepared from non-smoothed data. Analvte PFOS Tetra-H-PFOS (IS) Ifin m/z = 499.0 m/z = 427.0 Retention Time 3.1 2.9 ADVANCED B IO AN A LYTIC A L S E R V IC E S , INC. 3M Environmental Laboratory Page 35 of 50 99VDJA01.MI.DOC Page 206 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Figure 6: Mass Chromatograms of Calibration Standard 10 in Rat Serum Extract Containing PFOS (20 pg/mL) and the Internal Standard m/z = 427.0 3.52e5 cps I 90 - oi 80 70 60 - 50 40 - 30 - 20 - 10 1 12 Timi:, min 3 4 All quantitation results and figures were prepared from non-smoothed data. Analvte PFOS Tetra-H-PFOS (IS) Ion m/z = 499.0 m/z = 427.0 Retention Time 3.1 2.9 3M Environmental Laboratory ADVANCED B IO AN A LYTIC A L S E R V IC E S , INC. Page 36 of 50 99VDJA01.MI.DOC Page 207 3IM Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 9. APPENDIX A: QUANTITATION OF PERFLUOROOCTANESULFONATE (PFOS) IN RAT SERUM BY TURBO ION SPRAY LC/MS AUTHORS: David J. Anderson, M.S. Amie J. Prince, B.S. Holly D. Ross, M.S. TABLE OF CONTENTS A.1.0 Chemical Structure(s)........................................................................................ ..38 A.2.0 Specimen(s)................................................... 38 A.3.0 Assay Principle................. 38 A.4.0 Compounds...... ............... 38 A.5.0 Chemicals.................................................................. ..........^.............................38 A.6.0 Materials and Equipment..................................................................................... 39 A.7.0 Preparation of Solutions......................................................................................42 A.8.0 Preparation of Quality Control Samples..............................................................45 A.9.0 Liquid-liquid Extraction Procedure.................................................................... 46 A. 10.0 Instrument Conditions........................ 47 A. 11.0 Calculations....................................... 49 3M Environmental Laboratory ADVANCED B IO AN A LYTIC A L S E R V IC E S , INC. Page 37 of 50 99VDJA01.MI.DOC Page 208 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 A.1.0 CHEMICAL STRUCTURE^) OII CgFn-----S-----0 `K+ II O Potassium Perfluorooctanesulfonate MW =538 O C6Fi3-----C-----C-----S-----OH h2 o IH, IH, 2H, 2H-Perfluorooctane Sulfonic Acid MW = 428, Internal Standard A.2.0 SPECIMEN(S) This assay uses 50-pL aliquots of rat seruin. Rat serum samples are stored at -20 C. A.3.0 ASSAY PRINCIPLE PFOS and its internal standard, Tetra-H-PFOS, are extracted from rat serum samples (50 pL) using a liquid-liquid extraction procedure. The organic layer is evaporated to dryness and then reconstituted. An aliquot is then analyzed by turbo ion spray liquid chromatography/mass spectrometry (LC/MS) in the negative ion mode. A.4.0 COMPOUNDS PFOS: Lot# 171, 99.976% purity, 3M, Inc., Minneapolis, MN. Tetra-H-PFOS: Lot# 59909, 90% purity, 3M, .'Inc., Minneapolis, MN. A.5.0 CHEMICALS All chemicals may be substituted with that of an equivalent manufacturer and grade o f chemical. 3M Environmental Laboratory ADVANCED BIO AN A LYTIC A L S E R V IC E S , INC. Page 38 of 50 99VDJA01 .MI.DOC Page 209 3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 Acetonitrile Ammonium Acetate Ethyl Acetate Methanol Rhodapex mass calibration solution Sodium Bicarbonate Sodium Carbonate, Annhydrous Tetrabutylammonium Hydrogen Sulfate Water Rat Serum Rat Serum Cat# 015-4, Burdick & Jackson, Muskegon, MI 49442 Cat# 24,019-2, Aldrich, Milwaukee, WI 53201 Cat# 100-4, Burdick & Jackson, Muskegon, MI 49442 Cat# 230-4, Burdick & Jackson, Muskegon, MI 49442 A mixture of 0.11 mM trifluoroacetic acid (J.T. Baker, Phillipsburg, NJ 08865), 0.52 mM Rhodapex CO-436,1 mM ABEX EP110 and 1 mM ABEX EP-120 (all three supplied by Rhone-Poulenc, Inc., Cranbury, NJ 08512) in 1:1 methanol:water. Cat# 3509-01, J.T. Baker, Phillipsburg, NJ 08865 Cat# 3604-01, J.T. Baker, Phillipsburg, NJ 08865 HPLC Grade, Cat# J360-07, J.T. Baker, Phillipsburg, NJ 08865 High Purity (NANOpure), Bamstead Model D7331 Ultrapure Water System, Dubuque, IA 52001 . Lampire Biological Laboratories, Pipeiville, PA 18947 Harbin Bioproducts for Science, Inc., Indianapolis, IN 46229 A.6.0 MATERIALS AND EQUIPMENT Mass Spectrometer PE SCIEX API 365 atmospheric pressure ionization tandem triple quadmpole mass spectrometer equipped with TurboIonSprayTM interface, PE SCIEX, Concord, Ontario L4K4V8 Data system API Standard Software, MacQuan v 1.4, LC2Tune v 1.3, MacDAD v 1.3, Bundler v 1.3, Multiview v 1.3, 3M Environmental Laboratory ADVANCED B IO AN A LYTIC A L S E R V IC E S , INC. Page 39 of 50 99VDJA01.MI.DOC Page 210 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 HPLC Pump LC Pump Controller Autosampler HPLC Column Harvard syringe pump 11 Multi-tube vortexer Balance pH Meter Mechanical shaker NANOpure-UV water purification system Microbalance Micro weigh boats Weigh boats Sorvall RT-6000D refrigerated centrifuge Beckman GS6KR Refrigerated centrifuge and Sample Control v 1.3, on a Power Macintosh, PE SCIEX, Concord, Ontario L4K 4V8 Shimadzu LC-10AD pump, Shimadzu Co., Columbia, MD 21046 Shimadzu SCL-10A, Shimadzu Co., Columbia, MD 21046 Waters 717plus, Waters Associates, Millipore Corporation, Milford, MA 01757 Betasil C18, 5 pm p:irticle size, 2.1 x 50 mm, Cat #852055-701, Keystone Scientific, Inc., Bellefonte, PA 16823 Harvard Apparatus Inc., South Natick, MA 01760 Cat# 58816-115, VWR Scientific, West Chester, PA 19380 Model FX-300, ANfD Ltd., Tokyo, Japan Model 340, Coming Inc., Coming, NY 14830 Cat# 6000, Eberbach Corp., Ann Arbor, MI 48106 Bamstead, Dubuque, LA 52001 Model MT-5, Mettler-Toledo Inc., Hightstown, NJ 08520 Cat# 0219-0041, Perkin-Elmer Corp., Norwalk, CT 06859 Cat# 12577-025, VWR Scientific, West Chester, PA 19380 Cat# 83071, DuPont Co., Wilmington, DE 19898 Beckman, Palo Alto, CA 94304 I f -- M | | H ADVANCED b io a n a l y t ic a l S E R V IC E S . IN C. 3IVI Environmental Laboratory Page 40 o f 50 99VDJA01.MI.DOC Page 211 m Medical Department Study: T-6295.12 . Analytical Report: FACT TOX-110 LRN-U2849 TurboVap LV Evaporator Pipettes Pipette tips Polypropylene tubes Screw-capped Polypropylene vials Screw-capped Polypropylene vials Plug Tite Multipurpose Caps Pyrex volumetric flask Solvent Filtration Apparatus Nylon Titan Membrane Filters PEEK tubing Liquid Nitrogen Gastight Glass Syringes Cat# 43750, Zymark Instruments, Hopkinton, MA 01748 Cat# P-5000, P-1000, P-200, P-100, Rainin Instrument Co., Woburn, MA 01888 RT-20, RT-200, C-5000, CP-25, CP-50, CP-250, Rainin Instrument Co., Woburn, MA 01888 13 mm x 100 mm, Cat# 15070-574, VWR Scientific, West Chester, PA 19380 16.5 x 57 mm, Cat# 60.542, Sarstedt, Inc., Newton, SC 28658 16.5 x 101 mm, Cat# 60.541, Sarstedt, Inc., Newton, SC 28658 Cat# 78-127-0019-100, Elkay Products, Inc., Worcester, MA 01607 Cat# 28014P-10, 23014P-25, 28014P-100, 28014P1000, Kimble/Kontes, Vineland, NJ 08360 Cat# 953781-0000., 953751-0000, 953753-0000, 953826-0000, 953827-0000, Kimble/Kontes, Vineland, NJ 08360 0.45 pm Scientific Resources Inc., Cat# 74547-NN, Eatontown,NJ 07724 0.005" i.d. x 1/16" o.d., Cat# 1535, Upchurch Scientific Inc., Oak Harbor, WA 98277 Supplied in-house from bulk tank, BOC Gasses, Buffalo, NY 14210-2005 Cat# 1750, Hamilton Company, Reno, NV 89510 Other general laboratory glassware and supplies were used. I liB illM H B ADVANCED b io a n a l y t ic a l S E R V IC E S . IN C . 3M Environmental Laboratory Page 41 of SO 99VDJA0I MI.DOC Page 212 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 A.7.0 PREPARATION OF SOLUTIONS NANOpure water (18 megaohm-cm.) or equival ent should be used wherever water is called for. Mix all solutions well. A.7.1 Preparation of Analytical Standard Stock Solutions Analytical Standard Stock Solution, PFOS (1 mg/mL): Weigh approximately 5 mg of PFOS (after correction for purity) on a microbalance and transfer it to a polypropylene vial. Dilute with the appropriate volume of methanol to yield a 1 mg/mL solution. Prepare a fresh solution every three months. Store the solution at 4 C and bring to ambient temperature before use. Standard Spiking Solutions Prepare Standard Working Solutions A through J in 5-mL class "A" volumetric flasks according to the following dilution scheme. Dilute to the 5-mL mark with water to yield the final concentration. Prepare fresh solutions every three months. Store the solutions at 4 C and bring to ambient temperature before use. Standard Working A B C D E F G H I J Volume Spiked Solution Used (ML) 1000 Stock 800 Stock 400 Stock 200 Stock 100 Stock 250 200 pg/mL (A) 125 200 pg/mL (A) 62.5 200 pg/mL (A) 500 10 pg/mL (F) 250 10 pg/mL (F) PFOS Final Cone. (pg/mL) 200 . 160 80 40 20 10 5 2.5 1.0 0.5 ADVANCED B IO AN A LYTIC A L S E R V IC E S , INC. 3IVI Environmental Laboratory Page 42 of 50 99VDJA01 .MI.DOC Page 213 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 A.7.2 Preparation of Internal Standard Stock Solutions Internal Standard Stock Solution, Tetra-H-PFOS (1 mg/mL): Weigh approximately 5 mg of Tetra-H-PFOS on a microbalance and transfer it to a polypropylene vial. Dilute with an appropriate volume of methanol to yield a 1 mg/mL solution. Prepare a fresh solution every three months. Store the solution at 4 C and bring to ambient temperature before use. A.7.3 Preparation of Internal Standard Working Solution Internal Standard Working Solution (4 pg/mL Tetra-H-PFOS): Add 400 pL of Internal Standard Stock Solution to a 100-mL class "A" volumetric flask. Dilute to the mark with water to give a 4 pg/mL Tetra-H- PFOS solution. Store the solution at 4 C and bring to ambient temperature before use. Prepare fresh solution as needed. A.7.4 Preparation of Standard Curve and Control Blank Prepare fresh calibration standards for each analytical run by combining 360 pL of rat control serum and 40 pL of the analytical standard working solution (see table below) in labeled 1,7-mL microtubes. Prepare control blanks and zero samples by combining 360 pL of rat control serum with 40 pL of water. Vortex each for 60 seconds. 3M Environmental Laboratory ADVANCED BIO AN ALYTIG AL S E R V IC E S , INC. Page 43 of 50 99VDJA01.MI.DOC Page 214 3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 Analytical Standard Dilution Table: Standard # 10 9 8 7 6 5 4 3 2 1 Volume Spiked (pL) 40 40 40 40 40 40 40 40 40 40 Solution to use A B C D E F G H I J pL control serum 360 360 360 360 360 360 360 360 360 360 Final Cone. (pg/mL) 20 16 8 4 2 1 0.5 0.25 0.1 0.05 A.7.5 Preparation of Other Solutions 0.25 M Sodium Carbonate/0.25 M Sodium Bicarbonate: Weigh 26.5 g of sodium carbonate and 21 g of sodium bicarbonate and dissolve in approximately 900 mL of HPLC-grade water. Transfer the solution to a 1000-mL volumetric flask and dilute to the mark with HPLC-grade water. Mix the solution thoroughly. Store at ambient temperature. Prepare a fresh solution every three months. 1 M Ammonium Acetate: Add 7.7 g of ammonium acetate to a 100-mL volumetric flask, dissolve in NANOpure water, and stir until completely dissolved. Dilute to the mark with NANOpure water. Store at ambient temperature. Prepare a fresh solution every three months. 2 mM Ammonium Acetate: Combine 2 mL o f 1M ammonium acetate and 900 mL NANOpure water in a 1000-mL volumetric flask; stir until completely mixed. Dilute to the mark with NANOpure water. Store at ambient temperature. Prepare a fresh solution every three months. 3M Environmental Laboratory ADVANCED B IO AN A LYTIC A L S E R V IC E S . IN C . Page 44 of 50 99VDJA01.MI.DOC Page 215 31V! Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 10 M Sodium Hydroxide: Add 40 g of sodium hydroxide to a 100-mL volumetric flask, dissolve in NANOpure water, and stir until completely dissolved. Dilute to the mark with NANOpure water. Store at ambient temperature. Prepare a fresh solution every three months. 1 M Sodium Hydroxide: Add 10 mL of 10 M sodium hydroxide to a 100-mL volumetric flask. Dilute to the mark with NANOpure water; stir until completely mixed. Store at ambient temperature. Prepare a fresh solution every three months. 0.5 M Tetrabutylammonium Hydrogen Sulfate, pH 10: Weigh 16.98 g of tetrabutylammonium hydrogen sulfate and dissolve in 40 jnL o f NANOpure water. Adjusted the pH to 10.0 with 10 M and 1 M sodium hydroxide. Transfer the solution to a 100-mL volumetric flask and dilute to the mark with NANOpure water. 10:90 Methanol:2 mM Ammonium Acetate (Mobile Phase Eluent A): Add 100 mL of methanol and 900 mL of 2 mM ammonium acetate to a 1000-mL Pyrex bottle. Mix the solution thoroughly and filter the solution through a 0.45 pM filter with a vacuum flask. Store at ambient temperature. Prepare a fresh solution every three months. 90:10 Methanol:2 mM Ammonium Acetate (Mobile Phase Eluent B): Add 900 mL of methanol and 100 mL of 2 mM ammonium acetate to a 1000-mL Pyrex bottle. Mix the solution thoroughly and filter the solution through a 0.45 pM filter with a vacuum flask. Store at ambient temperature. Prepare a fresh solution every three months. A.8.0 PREPARATION OF QUALITY CONTROL SAMPLES A.8.1 Preparation of Quality Control Stock Solutions QC Stock Solution, PFOS (1 mg/mL): Weigh approximately 5 mg of PFOS (after correction for purity) on a microbalance and transfer it to polypropylene vial. Dilute 3IVI Environmental Laboratory ADVANCED BIO AN A LYTIC A L S E R V IC E S . INC. Page 45 of 50 99VDJA01.M1.DOC Page 216 3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 with the appropriate volume of methanol to yield a 1 mg/mL solution. Store the solution at 4 C and bring to ambient temperature before use. Prepare a fresh solution every three months. A.8.2 Preparation of Serum QC Samples QC4 (Dilution QC, 100 pg/mL PFOS): Add 1000 pL QC Stock Solution to a 10 mL class "A" volumetric flask to yield a 100-pij/mL solution. Dilute to the mark with control serum, cap and mix. Aliquots of approximately 0.2 mL are placed in polypropylene vials and stored frozen a t-20 C. QC3 (18 pg/mL PFOS): Add 450 pL QC Stock Solution to a 25-mL class "A" volumetric flask to yield a 18-pg/mL solution. Dilute to the mark with control serum, cap and mix. Aliquots of approximately 0.5 mL are placed in polypropylene vials and stored frozen at -20 C. QC2 (6 pg/mL PFOS): Add 150 pL of QC Stock Solution to a 25-mL class "A" volumetric flask to yield a 6-pg/mL solution. Dilute to the mark with control serum, cap and mix. Aliquots of approximately 0.5 ml. are placed in polypropylene vials and stored frozen at -20 C. QC1 (0.2 pg/mL PFOS): Add 111.1 pL of QC'3 to a 10-mL class "A" volumetric flask to yield a 0.2-pg/mL solution. Dilute to the mark with control serum, cap and mix. Aliquots of approximately 0.5 mL are placed in polypropylene vials and stored frozen at -20 C. A.9.0 LIQUID-LIQUID EXTRACTION PROCEDURE 1. Prepare fresh calibration standards for each, run according to Section A.7.4. 2. Thaw QC samples. 3. Prepare QC4 samples (diluted 1 in 10) in two steps: a) Add 180 pL of control serum and 20 pL of QC4 into 1.7-mL microtubes and 3IVI Environmental Laboratory ADVANCED B IO AN A LYTIC A L S E R V IC E S . INC. Page 46 of 50 99VDJA01.MI.DOC Page 217 3M Medical Department Study: T-6295.12 , Analytical Report: FACT TOX-110 LRN-U2849 vortex. . b) Aliquot 50 pL of diluted QC4 into labeled 16 x 100 mm polypropylene tubes. 4. Aliquot 50 pL of each control blank, zero sample, calibration standard, and quality control samples 1-3 into labeled 16 x 100 mm polypropylene tubes. Standards and blanks are analyzed in duplicate. All QC samples are analyzed in replicates of five. 5. Add 1 mL of 0.5 M tetrabutylammonium hydrogen sulfate, pH 10 to each tube. 6. Add 2 mL of 0.25 M sodium carbonate/0.25 M sodium bicarbonate to each tube. 7. Add 500 pL of the Internal Standard Working Solution to each tube (except control blank, add 500 pL water). 8. Add 5 mL ethyl acetate to each tube. 9. Shake the tubes on a reciprocal shaker at medium speed for 20 minutes. 10. Centrifuge the tubes at 3000 rpm for 20 minutes at 20 C. 11. Freeze the aqueous phase in an acetone/dry ice bath and keep the tubes in the bath an additional 5 minutes. 12. Transfer the ethyl acetate layer to a fresh, labeled 13 x 100 mm polypropylene tube. . 13. Evaporate the ethyl acetate layer to dryness in a TurboVapTM at approximately 20 C under nitrogen. 14. Reconstitute the dried extracts in two steps: Add 500 pL acetonitrile and vortex for 60 seconds. Then add 500 pL water and vortex for 60 seconds. 15. Transfer 200 pL of the extract to a labeled polypropylene autosampler vial.. A.10.0 INSTRUMENT CONDITIONS HPLC Conditions: Eluent (gradient) Mobile Phase A: 10:90 methanol:2mM ammonium acetate (v/v) Mobile Phase B : 90:10 methanol:2 mM ammonium acetate (v/v) ADVANCED B ID A N A LY TIC A L S E R V IC E S . IN C . 3M Environmental Laboratory Page 47 of 50 99VDJA01.MI.DOC Page 218 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Eluent Gradient Conditions: Time (min) 0 1.0 5 5.5 8 Flow Rate Autosampler Autosampler Temperature Injection Volume HPLC Column HPLC Column Temperature Typical Initial Column Pressure Autosampler Run Time %B 45 100 100 45% Stop 300 pL/min. Waters 717plus Ambient 3 to 5 pL Keystone Betasil C18, 5 pm particle size, 2.1 mm x 50 mm Ambient 90 bat: 8.0 minutes Analyte Reoresentative Retention Time (minutes) PFOS Tetra-H-PFOS (IS) - 3.1 2.9 Mass Snectrometer Conditions: Curtain Gas Nebulizer Gas TurboIonSprayTM Temperature TurboIonSprayTM Auxiliary Gas UHP Nitrogen UHP Nitrogen 400 C UHP Nitrogen at 8 L/min 3M Environmental Laboratory ADVANCED B IO AN A LYTIC A L S E R V IC E S , IN C . Page 48 of 50 99VDJA01.MI.DOC Page 219^ 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Ions Monitored: Analvte PFOS Tetra-H-PFOS (IS) Ion Monitored m/z = 499.0 m/z= 427.0 Dwell Time 500 ms 500 ms Ionization Mode Ion Spray Voltage Declustering Potential MS Acquisition Time Pause Time Negative Ion -3000 V -35 V 5 minutes 2 ms Calibrate the mass axis of the instrument by infusion of Rhodapex mass calibration solution (Section A.5.0) at a flow rate of 10 pL/min. Optimize the sensitivity of the instrument using an infusion of a 10 pg/mL solution of the analyte at 10 pL/min into a flow of 190 pL/min of mobile phase using the: gradient condition at which the analyte elutes from the LC column (100% eluent B). Mass spectral peak widths should be approximately 0.6 amu at half-height. One set of calibration standards was injected at the beginning and one set of calibration standards was injected at the end of each analytical run (tray). A.11.0 CALCULATIONS Calculated concentrations are based on peak area ratios of PFOS to Tetra-H-PFOS. The peak area for the analyte ion is divided by the peak area for the corresponding internal standard ion. Data generated from samples in this study were acquired and integrated using PE SCIEX software applications Sample Control (v. 1.3) and MacQuan (v. 1.4). The validation data were formatted in the ABS laboratory information management system (Watson, v.5.3.1.01). All data were rounded to no less than three significant figures by ABS prior to reporting. ADVANCED B ID A N A LY TIC A L S E R V IC E S , INC. 3IVI Environmental Laboratory Page 49 of 50 99VDJA01.MI.DOC Page 220 3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 Acceptance Criteria for Sample Analysis Calibration standards: The coefficient of determination (r2) must be >0.98. At least three-fourths of the individual calibration standard replicates will have deviations within 15% from their nominal concentrations. LLQ acceptance criteria: At least one replicate at the LLQ must exhibit a deviation within 20% from its nominal concentration. If this criterion is not met, both replicates are rejected and the standards at the next higher level are subjected to the same test. Quality control samples: At least two-thirds of the individual QC sample replicates will have deviations within 15% from their nominal concentrations, with at least one replicate at each QC concentration meeting this criterion. 3M Environmental Laboratory ADVANCED BIO AN A LYTIC A L S E R V IC E S , INC. Page 50 of 50 99VDJA01.MI.DCX; Page 221 3M Medical Department Study: T-6295.12 ADVANCED BIOANALYTICAL SERVICES, INC. BIOANALYTICAL REPORT Analytical Report: FACT TOX-110 LRN-U2849 Title: ^ Date: Quantitative Determination of Perfluorooctanesulfonate (PFOS) in Rat Serum from Study #FACT-TOX-l 10 Using Turbo Ion Spray LC'/MS 21 September 1999 Authors: David J. Anderson, M.S. Amie J. Prince, B.S. Holly D. Ross, M.S. Prepared For: 3M Environmental Technology and Safety Services St. Paul, MN 55133-3331 ABS Report No.: 99ADJA02.MI.DOC Study Protocol: FACT-TOX-110 Number of Pages: 24 ** Sxunmary and Conclusions The concentration of perfluorooctanesulfonate (PFOS) was determined in rat serum samples collected during the study #FACT-TOX-l 10 using a sensitive, specific, accurate, and reproducible analytical method developed by Advanced BioAnalytical Services, Inc., *' Ithaca, NY. Rat serum samples (50 pL) were extracted by a liqui d-liquid extraction procedure to isolate PFOS and the internal standard, lH,lH,2H,2H-perfluorooctane sulfonic acid (Tetra-H-PFOS). Following evaporation and reconstitution, sample extracts were analyzed by turbo ion spray liquid chromatography/mass spectrometry (LC/MS) in the negative ion mode. All samples were successfully analyzed within six runs. The lower limit of quantitation was 0.05 pg/mL for PFOS. The precision of this assay (RSD) as determined from the ^ analysis of quality control samples for PFOS was <6.90%. The precision of this assay (RSD), as determined from the calibration standards for PFOS was <6.18%. The relative error (RE) o f the assay, as determined from the analysis of the quality control samples ranged from 1.52 to 12.4%. The RE of the assay, as determined from the analysis of calibration standards ranged from -4.62 to 10.3%. 15 C atherw ood Road Ith aca, New York 14850 (607) 266-0665 Fax (607) 266-0749 3M Environmental Laboratory Page 222 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Title: ADVANCED BIOANALY'OCAL SERVICES SIGNATURE PAGE Quantitative Determination of Perfluorooctanesulfonate (PFOS) in Rat Serum from Study #FACT-TOX-l 10 Using Turbo Ion Spray LC/MS Reviewed by: Authorized for Release by: Melissa Mapes, A.S. ' Associate Auditor fCi-- JohnlR. Perkins, Ph.D. Assistant Scientific Director Z\ S& ^ Date nserr) Date 1. fr f- l Date ADVANCED B IO AN A LYTIC A L S E R V IC E S , IN C . 3M Environmental Laboratory Page 2 of 24 99ADJA02.MI.DOC Page 223 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 QAU STATEBIENT Periodic inspections of the bioanalytical portion of study #FACT-TOX-l 10 were conducted by the Quality Assurance Unit of Advanced BioAnalytical Services (ABS) for compliance with EPA GLP regulations (40 CFR Part 792). The study was inspected on the following dates: 25 June 1999; 6, 7, 8, 21 July 1999; 31 August 1999. Results o f the inspection were reported to ABS Management and Project Team Leader on: 25 June 1999; 6, 7, 8, 21 July 1999; 31 August 1999. Results o f the inspections were reported to the Study Director on 2 September 1999. Based on the inspections and the data reviewed, this report is a complete and accurate representation of the data. Melissa Mapes, A.S. ' Associate Auditor Date M 1 ADVANCED B IO AN A LYTIC A L S E R V IC E S , IN C. 3M Environmental Laboratory Page 3 of 24 99ADJA02.MI.DOC Page 224 3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 TABLE OF CONTENTS SIGNATURE PAGE............................................................................................................ 2 QAU STATEMENT............................................................................................................. 3 TABLE OF CONTENTS...................................................................................................... 4 LIST OF TABLES................................................................................................................ 5 1. INTRODUCTION........................................................................................................ 6 1.1. Study Description and Objective.............................................................................. 6 2. METHODS................................................................................................................... 6 2.1. Analytical Procedure(s)................................ 6 2.2. Assay Site..................................................... 7 2.3. Data Processing............................................ 7 3. RESULTS.................................................................... 7 3.1. Assay Performance................................................................................................... 7 3.2. Analytical Results for Study #FACT-TOX-110.......................................................8 4. SUMMARY AND CONCLUSIONS...........................................................................8 5. DATA RETRIEVAL.................................................................................................... 8 6. REFERENCES............................................................................................................. 8 7. TABLES....................................................................................................................... 9 ADVANCED B ID A N A LV TIC A L S E R V IC E S , IN C . 3IVI Environmental Laboratory Page 4 of 24 99ADJA02.MI.DOC Page 225 3M Medical Department Study. T-6295.12 r* Analytical Report: FACT TOX-110 LRN-U2849 LIST OF TABLES Table 1: Summary of Sample and Assay Information for Protocol FACT-TOX-110..... 9 Table 2: Inter-Assay Precision and Accuracy for PFOS Quality Control Samples in Rat Serum for Study #FACT-TOX-l 10................................................................. 10 Table 3: Inter-Assay Precision and Accuracy for PFOS Calibration Standards in Rat Serum for Study #FACT-TOX-l 10..................................................................11 Table 4: Calibration Curve Statistics for the Determination of PFOS in Rat Serum for Study #FACT-TOX-110....................................................................................12 Table 5: Concentrations of PFOS in Rat Serum Simples from Study #FACT-TOX-l 10 Following the 0 mg/kg Dose................. ...........................................................13 Table 6: Concentrations of PFOS in Rat Serum Simples from Study #FACT-TOX-l 10 Following the 0.1 mg/kg Dose............................................................... ..........16 Table 7: Concentrations of PFOS in Rat Serum Simples from Study #FACT-TOX-l 10 Following the 0.4 mg/kg Dose.......................................................................... 18 Table 8: Concentrations of PFOS in Rat Serum Simples from Study #FACT-TOX-l 10 Following the 1.6 mg/kg Dose.......................................................................... 20 Table 9: Concentrations of PFOS in Rat Serum Simples from Study #FACT-TOX-l 10 Following the 3.2 mg/kg Dose.......................................................................... 22 Table 10: Repeat Analysis of Rat Serum Samples from Study #FACT-TOX-l 10......... 24 ADVANCED B IO AN A LYTIC A L S E R V IC E S . IN C . 3M Environmental Laboratory Page 5 of 24 99ADJA02.MI.DOC Page 226 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 1. INTRODUCTION 1.1. Study Description and Objective The objective of this study was to determine the concentrations of perfluorooctanesulfonate (PFOS) in rat serum samples collected during the study #FACTTOX-110, "Oral (Gavage) Pharmacokinetic Study of PFOS in Rats." Rat serum samples were analyzed using a liquid-liquid extraction and turbo ion spray liquid chromatography/mass spectrometry (LC/MS) assay. 2. METHODS 2.1. Analytical Procedure(s) PFOS concentrations were determined in rat serum according to the validated LC/MS method (1). Serum samples (50 pL) were extracted by a liquid-liquid extraction procedure using ethyl acetate to isolate PFOS and the internal standard (IS), lH,lH,2H,2H-perfluorooctane sulfonic acid (Tetra-H-PFOS) from rat serum. Following evaporation to dryness and subsequent reconstitution, sample extracts were separated by reversed-phase chromatography on a 2 x 50 mm (5 pm) BetasilTM C18column (Keystone Scientific, Inc., Bellefonte, PA) with an initial mobile phase of 55% Eluent A (10:90 methanol:2 mM ammonium acetate) and 45% Eluent B (90:10 methanol:2 mM ammonium acetate). PFOS concentrations were determined by turbo ion spray liquid chromatography/mass spectrometry (LC/MS) in selected ion monitoring (SIM) mode. The negative ions monitored were: m/z = 499.0 for PFOS [M-K]' m/z = 427.0 for Tetra-H-PFOS [M-H]" Study sample concentrations were determined from a weighted (1/y2), quadratic regression of peak area ratios (peak area of PFOS/peak area of Tetra-H-PFOS) versus nominal concentration of ten calibration standards. The nominal concentrations of the calibration standards were 0.05,0.1,0.25, 0 .5,1,2,4, 8,16, and 20 pg/mL PFOS. The lower limit of quantitation (LLQ) for this assay was determined to be 0.05 pg/mL PFOS. Quality control (QC) serum samples at three different concentrations (0.2, 6, and 18 pg/mL PFOS) were analyzed with each assay batch in replicates of four. Dilution QC samples (Q C4,100 pg/mL) were prepared at each dilution that was used during a particular assay and were analyzed with each assay batch in replicates of four as needed. The acceptance criteria for calibration standards stipulated that the back-calculated concentrations of at least three-fourths of the individual calibration standard replicates must not deviate more than 15% from their nominal concentration (except at the LLQ). At least one duplicate at the LLQ must exhibit a deviation within 20% of the nominal ADVANCED BIO AN A LYTIC A L S E R V IC E S , IN C. 3M Environmental Laboratory Page 6 of 24 99ADJA02.MI.DOC Page 227 3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 concentration . The coefficient of determination (r2) must be >0.98. The acceptance criteria followed are in accordance with ABS Standard Operating Procedures. The acceptance criteria for the quality control samples stipulated that at least two-thirds of the individual QC sample replicates must not deviate more than 15% from their nominal concentrations. At least one replicate at each QC concentration must exhibit a deviation within 15%. A summaiy of the sample and assay information for study #FACT-TOX-l 10 is given in Table 1. 2.2. Assay Site All samples were analyzed at Advanced BioAnalytical Services, Inc., Ithaca, NY. 2.3. Data Processing The data were collected using turbo ion spray LC/MS selected ion monitoring (SIM) in the negative ion mode. Peak areas were integrated by the PE SCIEX program MacQuan, version 1.4, residing on a Macintosh computer. Following peak area integration, the results tables from MacQuan were saved as text files and uploaded to the ABS file server where a weighted (1/y2) quadratic regression was performed using the software package Watson (v 5.3.1.01 PSS, Inc., Wayne, PA 19087). Calculations were performed on unrounded numbers. All calibration standard and Q'C results were rounded to no less than three significant figures before reporting. 3. RESULTS 3.1. Assay Performance The performance of the assay for PFOS as determined from the analysis of daily quality control samples is documented in Table 2. The inter-assay precision (RSD) of quality control samples ranged from 2.37 to 6.90% for PFOS. There was no marked inaccuracy in the results from these quality control samples; the relative error (RE) ranged from 1.52 to 12.4%. The performance of daily calibration curves is documented in Table 3. The inter-assay precision (RSD) of the standards ranged from 1.70 to 6.18% for PFOS. There was no marked inaccuracy in the results from these standards; the relative error (RE) ranged between -4.62 to 10.3%. ADVANCED B IO AN A LYTIC A L S E R V IC E S . INC. 3M Environmental Laboratory Page 7 of 24 99ADJA02.MI.DOC Page 228 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 The slope, y-intercept, and coefficient of determination (r2) for each analytical run are presented in Table 4. The r2values ranged from 0.9955 to 0.9965 for PFOS in rat serum. All ABS personnel assigned to analyze samples for this project were required to successfully extract quality control (QC) samples 2nd calibration standards. Results of the extraction were reviewed by a trained analyst against the pre-defined acceptance criteria for this assay. Results are maintained in the study records of ABS. 3.2. Analytical Results for Study #FACT-TOX-110 PFOS concentrations in rat serum from Study #FACT-TOX-l 10 are presented in Tables 5 through 9. All data were rounded to no less than tliree significant figures before reporting in Tables 5 through 9. Study samples requiring repeat analysis for Study #FACT-TOX110 are presented in Table 10. 4. SUMMARY AND CONCLUSIONS The concentration of PFOS was determined in rat serum samples collected during the study #FACT-TOX-l 10. PFOS was isolated from serum by a liquid-liquid extraction procedure and determined by LC/MS. The quality of the determinations was satisfactory throughout. The LLQ was 0.05 pg/mL PFOS. The precision of the assay, as determined from the analysis of quality control samples was <6.90% for PFOS. 5. DATA RETRIEVAL The calculated concentration data from the analysis of rat serum samples for Study #FACT-TOX-l 10 are maintained on file in the archives of the Advanced BioAnalytical Services, Inc., Ithaca, NY in ABS Notebook 2324. 6. REFERENCES 1. Advanced BioAnalytical Services Validation Report 99VDJA01 .MI.DOC. Method Validation for the Quantitation of Perfluorooctanesulfonate (PFOS) in Rat Serum by Turbo Ion Spray LC/MS. David J. Anderson, M.S. Amie J. Prince, B.S., and Holly D. Ross, M.S. 26 August 1999. ADVANCED B IO AN A LYTIC A L S E R V IC E S , IN C. 3M Environmental Laboratory Page 8 o f 24 99ADJA02.MI.DOC Page 229 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 7. TABLES Table 1: Summary of Sample and Assay Information for Protocol FACT-TOX110 Species/Matrix: Rat/Serum Sample Collection and Storage Information: Anticoagulant/Stabilizer: Reported Sample Collection Dates: Dates Received at ABS: Storage Temperature at ABS: None 27 December 1998 to 22 January 1999 16 June 1999 -20 C Assay Information: Assay Period: 29 June 1999 to 8 July 1999 Analyte: Potassium perfluorooctanesulfonate (PFOS) Analytical Standard: Lot No: Source: PFOS 171 3M Internal Standard: 1H,1H,2H,2H- /> perfluorooctane sulfonic acid (Tetra-H-PFOS) Lot No: 59909 Source: 3M Calibration Range: Regression Method: Weighting Factor: Lower Limit of Quantitation: Upper Limit of Quantitation: quadratic i/y2 0.05 pg/mL 20 pg/mL ADVANCED B IO AN A LYTIC A L S E R V IC E S . INC. 3M Environmental Laboratory Page 9 of 24 99ADJA02.MI.DOC Page 230 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Table 2: Inter-Assay Precision and Accuracy for PFOS Quality Control Samples in Rat Serum for Study #FACT-TOX-110 PFOS Concentration (ug/mL) Run No. QC1 QC2 QC3 QC4 QC4 QC4 0.2 6 18 100 100 100 (1 in 10 Dilution) (1 in 20 Dilution) (1 in 30 Dilution) 1 0.178 6.18 15.9 NA NA NA 0.183 6.40 17.3 NA NA NA 0.177 6.32 17.3 NA NA NA 0.187 6.32 17.3 NA NA NA 3 0.210 6.24 17.9 NA NA NA 0.215 5.79 17.2 NA NA NA 0.211 5.99 17.8 NA NA NA 0.211 5.87 17.9 NA NA NA 4 0.225 6.14 18.1 116* NA NA 0.221 6.09 18.4 110 NA NA 0.222 6.11 18.2 110 NA NA 0.220 6.19 18.2 113 NA NA 5 0.220 6.14 19.0 0.218 6.31 18.1 0.218 6.27 18.5 0.214 6.42 18.7 NA NA NA NA 116* NA 112 NA 107 NA 114 NA 6 0.225 6.22 19.1 0.222 6.15 19.5 0.218 6.39 19.8 0.219 6.27 19.3 NA NA NA NA 108 NA 111 NA 111 NA 111 NA 7 0.221 6.11 18.6 NA NA 106 0.212 6.15 19.2 NA NA 104 0.215 6.17 18.5 NA NA 100 0.218 6.80 18.8 NA NA 100 Mean 0.212 6.21 18.3 112 111 103 RSD (%) 6.90 3.17 4.84 2.37 2.55 2.73 RE 5.80 3.50 1.52 12.4 11.2 2.54 RSD = (SD/Mean) x 100% RE = [(Mean-Nominal)/Nominal] x 100% NA: Not applicable "Individual replicate exceeds 15% acceptance criterion. ADVANCED B IO AN A LVTIC A L S E R V IC E S . IN C . 3M Environmental Laboratory Page 10 of 24 99ADJA02.MI.DOC Page 231 3M Medical Department Study: T-6295.12 ADVANCED BIO AN A LYTIC A L S E R V IC E S . IN C. 3IVI Environmental Laboratory Table 3: Inter-Assay Precision and Accuracy for PFOS Calibration Standards in Rat Serum for Study #FACT-TOX-110 Run No. STD1 STD2 STD3 0.05 0.1 0.25 - CO 0.0473 0.0981 0.246 0.0552 0.0985 0.261 0.0467 0.0985 0.237 0.0545 0.108 0.251 001'0 8600 801'0 eoro 0.0521 0.231 0.0474 0.241 TCo--Mo0l-H0 dd m so r- 0.0499 0.0526 0.234 0.250 0.0464 0.0530 0.252 0.248 Mean RSD (%) RE 0.0501 1 0.100 0.0528 0.0905 0.0507 0.101 6.18 5.05 1.33 1.05 0.266 .256 0.248 4.35 -0.887 RSD = (SD/Mean) x 100% RE = [(Mean-Nominal)/Nominal] x 100% PFOS Concentration (ug/mL) STD4 STD5 STD6 STD7 0.5 1 2 4 0.466 0.888 2.06 4.41 0.507 0.997 2.11 4.48 0.488 1.00 2.08 4.26 0.448 0.924 2.21 4.49 0.498 0.975 2.16 4.22 0.500 0.928 2.07 4.52 0.450 0.944 2.22 4.44 0.489 0.989 2.20 4.38 60'Z LYZ 0.473 0.953 4.42 0.471 0.938 4.30 0.476 0.958 2.15 4.54 0.478 0.948 2.00 4.47 0.479 0.954 4.41 3.89 3.52 2.32 -4.27 -4.62 10.3 ^m ^N n*--i cm rn so 7.83 7.62 7.90 7.86 7.91 7.64 7.83 1.80 -2.14 8 8G1S oOs sino 0t-0* ooo oo or* 00 00 91 6CLLS 16.5 15.5 15.1 15.7 15.9 15.7 15.8 15.4 16.0 15.5 16.1 15.8 15.8 2.28 -1.51 STD10 20 19.8 20.0 20.0 20.5 20.6 19.7 1 20.0 20.2 20.6 19.7 20.2 19.7 20.1 1.70 0.467 Analytical Report: FACTTOX-110 LRN-U2849 Page 232 Page 11 of 24 99ADJA02.MI.DOC 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Table 4: Calibration Curve Statistics for the Determination of PFOS in Rat Serum for Study #FACT-TOX-l 10 Run No. Quadratic Linear Coefficient (A) Coefficient (B) Intercept (C) 1 -0.0009199 0.1354 3 -0.0005189 0.1328 4 -0.0005052 0.1334 5 -0.0003195 0.1137 6 -0.0004256 0.1119 7 -0.0003264 0.1117 Mean -0.0005026 0.1232 y = Ax2+ Bx + C, weighted 1/y2 0.001580 0.002324 0.0004498 0.0009957 0.0001798 0.0001116 0.0009401 Coefficient of Determination (r2) 0.9960 0.9955 0.9965 0.9958 0.9963 0.9959 0.9960 s BIOANALYTIC AL S E R V IC E S . INC. 31VI Environmental Laboratory Page 12 of 24 99ADJA02.MI.DOC Page 233 3M Medical Department Study: T-6295.12 I )1 Table 5: Concentrations of PFOS in Rat Serum Samples from Study #FACT-TOX-110 Following the 0 mg/kg Dose Sampling Time (day) 19501 19502 19503 19504 PFOS Concentration (pg/mL) Rat Number* 19504F 19505 19505F 19506 19506F 19507 0 0.112 0.0701 0.0607 0.0514 NS 0.0550 NS 0.309 NS 0.0545 7 0.397 0.0535 NS 0.0926 NS 0.0514 NS 0.219 NS 0.0586 15 0.0803 0.0690 NS <LLQ[0.0432]<(0.05) NS 0.0609 NS 0.199 NS 0.0648 21 NS NS NS <LLQ(0.0258]<(0.05) 0.0845 <LLQ[0.0353]<(0.05) 0.0880 0.0820 0.267 <LLQ[0.0247]<(0.05) <LLQ = Less than the Lower Limit of Quantitation (0.05 pg/mL). Rat Number with "F" indicates fetal sample. NS: No sample. , Analytical Report: FACT TOX-110 LRN-U2849 Page 13 o f 24 99ADJA02.MI.DOC 3M Medical Department Study: T-6295.12 >I / r >I ) 1 Table 5: (Continued) Concentrations of PFOS in Rat Serum Samples from Study #FACT-TOX-110 Following the 0 mg/kg Dose Sampling Time (day) 19507F 19508 PFOS Concentration (pg/mL) Rat Number* 19509 19509F 19510 19511 19512 19512F 0 NS 0.0548 <LLQ[0.0483]<(0.05) NS 0.0664 0.0619 <LLQ[0.0449]<(0.05) NS 7 NS <LLQ[0.0395]<(0.05) <LLQ[0.0481]<(0.05) NS 0.0636 0.250 0.197 NS 15 NS <LLQ[0.0454]<(0.05) <LLQ[0.0458]<(0.05) NS 0.0614 0.176 0.238 NS 21 0.0842 NS <LLQ[0.0269]<(0.05) 0.102 NS NS 0.0833 0.219 <LLQ = Less than the Lower Limit of Quantitation (0.05 pg/mL). *Rat Number with "F" indicates fetal sample. NS: No sample. ,. Analytical Report: FACT TOX-110 LRN-U2849 Page 14 o f 24 99ADJA02.MI.DOC 3IVI Medical Department Study: T-6295.12 3M Environmental Laboratory ?>>>>) ) 1 01iD> UlnD 3j< o>q SSs 70 Table 5: (Continued) Concentrations of PFOS in Rat Serum Samples from Study #FACT-TOX-110 Following the 0 mg/kg Dose Sampling Time (day) 19513 PFOS Concentration (ng/mL)____________________________ __________ Rat Number*___________________________________ 19514 19515 19516_______ 19516F 0 0.0527 <LLQ[0.0500]<(0.05) <LLQ[0.0443]<(0.05) 0.0559 NS 7 <LLQ[0.0345]<(0.05) 0.0784 <LLQ[0.0488]<(0.05) <LLQ[0.0447]<(0.05) NS 15 <LLQ[0.0462]<(0.05) 0.0648 0.0581 <LLQ[0.0328]<(0.05) NS 21 NS NS NS <LLQ[0.0206]<(0.05) 0.165 <LLQ = Less than the Lower Limit of Quantitation (0.05 pg/mL). *Rat Number with "F" indicates fetal sample. NS: No sample. , Analytical Report: FACT TOX-110 LRN-U2849 Page 15 o f 24 99ADJA02.MI.DOC 3M Medical Department Study. T-6295.12 )I /l i )1 Table 6: Concentrations of PFOS in Rat Serum Samples from Study #FACT-TOX-110 Following the 0.1 mg/kg Dose Sampling Time (day) 19517 PFOS Concentration (pg/mL) Rat Number* 19518 19518F 19519 19519F 19520 19521 19521F 19522 19523 19523F 19524 0 9.07 9.87 NS 9.37 NS 9.73 11.1 NS 9.32 12.8 NS 10.2 7 8.63 10.3 NS 6.83 NS NS 8.82 NS 8.19 9.65 NS 10.4 15 11.5 10.7 NS 9.18 NS NS 14.1 NS 9.57 10.9 NS 9.20 21 NS 6.84 12.1 3.93 8.52 NS 4.93 11.2 NS 5.35 10.2 4.11 *Rat Number with "F" indicates fetal sample. NS: No sample. . Analytical Report: FACT TOX-110 LRN-U2849 Page 16 o f 24 99ADJA02.MI.DOC 3M Medical Department Study: T-6295.12 3M Environmental Laboratory ) l >t >i ) ) >) } tmn nm> n >nl i t m c-<ni .m HO 70 Table 6: (Continued) Concentrations of PFOS in Rat Serum Samples from Study #FACT-TOX-110 Following the 0.1 mg/kg Dose Sampling Time (day) 19524F 19525 19526 PFOS Concentration (pg/mL) Rat Number* 19527 19528 19528F 19529 19529F 19530 19531 19532 0 NS 12.0 9.48 12.0 10.0 NS 11.6 NS 8.48 9.10 10.6 7 NS 9.62 11.2 8.78 7.80 NS 10.3 NS 7.65 7.57 10.1 15 NS 10.0 11.7 8.78 8.04 NS 7.31 NS 11.8 10.4 9.66 21 10.3 NS NS NS 6.19 10.8 3.04 10.4 NS NS NS *Rat Number with "F" indicates fetal sample. NS: No sample. ,, Analytical Report: FACT TOX-110 LRN-U2849 Page 238 Page 17 of 24 99ADJA02.MI.DOC 3M Medical Department Study: T-6295.12 3M Environmental Laboratory >> > t ) > ) } ) ) 1 Table 7: Concentrations of PFOS in Rat Serum Samples from Study #FACT-TOX-110 Following the 0.4 mg/kg Dose COCD> miln l .HO 7n PFOS Concentration (pg/mL) Sampling Time Rat Number* (day) 19533 19533F 19534 19534F 19535 19536 19536F 19537 19538 19539 19540 19541 0 53.9 NS 46.9 NS 50.1 38.6 NS 56.6 46.5 45.2 42.9 44.4 7 44.3 NS 50.0 NS 56.6 36.6 NS 58.4 48.1 44.6 45.8 NS 15 47.3 NS 56.1 NS 48.2 43.3 NS 50.7 53.5 37.2 48.6 NS 21 20.4 42.5 20.5 41.4 NS 16.2 32.2 NS NS NS NS NS *Rat Number with "F" indicates fetal sample. NS: No sample. t Analytical Report: FACT TOX-110 LRN-U2849 P age 239 Page 18 of 24 99ADJA02.MI.DOC 3M Medical Department Study: T-6295.12 3M Environmental Laboratory Table 7: CDUJ > mn o n>i l l CmD[H<; mDn 7n at (day) 0 7 15 21 (Continued) Concentrations of PFOS in Rat Serum Samples from Study #FACT-TOX-110 Following the 0.4 mg/kg Dose PFOS Concentration (ng/mL) Rat Number* 19542 19543 19543F 19544 19545 19546 19546F 19547 19548 45.3 46.3 NS 45.8 47.9 43.5 40.6 46.1 NS 42.8 NS 46.5 44.0 43.9 NS 49.1 NS 47.3 NS 23.0 34.1 NS NS 35.8 NS 42.2 57.6 NS 42.2 60.0 NS 43.3 58.0 48.4 NS 65.7 *Rat Number with "F" indicates fetal sample. NS: No sample. .. ' Analytical Report: FACT TOX-110 LRN-U2849 Page 240 Page 19 of 24 99ADJA02.MI.DOC 3M Medical Department Study: T-6295.12 3M Environmental Laboratory -9 W } ) l ) )) Table 8: Concentrations of PFOS in Rat Serum Samples from Study #FACT-TOX-l 10 Following the 1.6 mg/kg Dose CD CD > 5g| S z rH dn> r Sampling Time (day) 19549 19550 0 166 200 7 NS 216 15 NS 160 21 NS NS 19551 PFOS Concentration (pg/mL) Rat Number 19552 19553 19554 19555 19556 19557 19558 19559 19560 184 165 182 203 166 199 198 196 171 200 NS NS 160 189 195 186 NS 171 160 171 NS NS 135 176 225 202 NS 164 192 215 NS NS NS NS NS 251 NS NS 237 79.4 NS: No sample. ,, Analytical Report: FACT TOX-110 LRN-U2849 T) Page 20 of 24 99ADJA02.MI.DOC <CQnD) ho 3M Medical Department Study: T-6295.12 3M Environmental Laboratory >> # > I ) > ) ) ) ) cnm> mn ni l>ln ?H O n Oi Table 8: (Continued) Concentrations of PFOS in Rat Serum Samples from Study #FACT-TOX-110 Following the 1.6 mg/kg Dose Sampling Time (day) 19560F 19561 PFOS Concentration (pg/mL) Rat Number3" 19562 19562F 19563 0 NS 193 185 NS 163 7 NS 181 179 NS 168 15 NS 219 142 NS 183 21 131 NS 63.7 102 NS 19564 185 169 159 NS *Rat Number with "F" indicates fetal sample. NS: No sample. t Analytical Report: FACT TOX-110 LRN-U2849 Pa9e 242 Page 21 of 24 99ADJA02.MI.DOC 3M Medical Department Study: T-6295.12 31V Environmental Laboratory \J )J >J )) ) Table 9: Concentrations of PFOS in Rat Serum Samples from Study #FACT-TOX-l 10 Following the 3.2 mg/kg Dose CO(D> milnOl nP d S 7 Sampling Time (day) 19565 19566 0 370 337 7 392 280 15 345 311 21 NS NS 19567 366 327 294 NS PFOS Concentration (gg/mL) Rat Number* 19568 19568F 19569 19569F 19570 19571 19571F 19572 19572F 339 NS 334 NS 351 348 NS 357 NS 340 NS 323 NS 337 371 NS 305 NS 336 NS 314 NS 269 308 NS 317 NS 241 228 142 176 NS 165 166 123 166 *Rat Number with "F" indicates fetal sample. NS: No sample. i ! Analytical Report: FACT TOX-110 LRN-U2849 Page 22 of 24 99ADJA02.MI.DOC 3M Medical Department Study: T-6295.12 ) > >J > ) J ) ) J > [cnf]mo>9 i n l > ln 53 P Table 9: (Continued) Concentrations of PFOS in Rat Serum Samples from Study #FACT-TOX-110 Following the 3.2 mg/kg Dose Sampling Time (day) 19573 0 364 7 343 15 317 21 NS 19574 365 393 299 NS 19575 413 387 284 NS PFOS Concentration (pg/mL) Rat Number* 19576 19577 19577F 19578 19579 19579F 19580 383 417 NS 395 NS 361 NS 221 NS 373 387 NS 378 NS NS 399 NS 357 NS NS 386 NS 323 226 NS 186 181 NS *Rat Number with "F" indicates fetal sample. NS: No sample. ,. Analytical Report: FACT TOX-110 LRN-U2849 Page 23 o f 24 99ADJA02.MI.DOC 3M Medical Department Study: T-6295.12 rntal Laboratory ) > 1 .W > > ) ) ) ) mcnnro > Ss Table 10: Repeat Analysis of Rat Serum Samples from Study #FACT-TOX-110 Rat Treatment Time Original Number Cone. pg/ml 19565 5 Day 7 >ULQ-428.6743>(400) 19565 5 Day 15 >ULQ-440.2439>(400) 19567 5 Day 0 >ULQ-402.0010>(400) 19574 5 Day 7 >ULQ-404.8033>(400) 19575 5 Day 0 >ULQ-436.6951>(400) 19577 5 Day 0 >ULQ-407.5918>(400) 19579 5 Day 0 >ULQ-403.4253>(400) >ULQ: Greater than the Upper Limit of Quantitation. REASONS FOR REASSAY: 1). Run 6 concentration exceeded calibration range. Original Reason Curve for Number Reassay 61 61 61 61 61 61 61 .IT,' : Reassay Cone. pg/ml 392 345 366 393 413 417 387 Reassay Curve Number 7 7 7 7 7 7 7 Reported Cone. pg/ml 392 345 366 393 413 417 387 Reason for Reported Cone. 1 1 1 1 1 1 1 REASONS FOR REPORTED CONC: 1). Run 7 concentration within calibration range. ,. Analytical Report: FACT TOX-110 LRN-U2849 Page 245 Page 24 of 24 99ADJA02.MI.DOC 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 ADVANCED m m iam mW i W P W B IO A N A LV T IC A L s e r v ic e s , in c . ADDENDUM TO THE BIOANALYTICAL REPORT Title: Addendum - Quantitative Determination of Perfluorooctanesulfonate (PFOS) in Rat Serum from Study #FACT-TOX-110 Using Turbo Ion Spray LC/MS Date: 13 October 1999 Addendum Report: 99ADJA02.MI.ADD.DOC Original Report: 99ADJA02.MI.DOC K Authors: David J. Anderson, M.S. Amie J. Prince, B.S. Holly D. Ross, M.S. Prepared For: 3M Environmental Technology and Safety Services S t Paul, MN 55133-3331 Number of Pages: 3 /m* ^ 15 Ca therwood Road Ithaca, New York 14850 (607) 266-0665 Fax (607) 266-0749 3M hnvironmental Laboratory Page 246 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 ADVANCED BIOANAIATTICAL SERVICES Title: SIGNATURE PAGE Addendum - Quantitative Determination of Perfluorooctanesulfonate (PFOS) in Rat Serum from Study #FACT-TOX-110 Using Turbo Ion Spray LC/MS ABS Report No.: 99ADJA02.MI. .DOC Reported by: David J. Anderson, M.S. Research Scientist 13 her Date Reviewed by: dathleen Cormack, B.S.. Associate Auditor Authorized for Release by: John R. Perkins, Ph.D. Assistant Scientific Director Accepted by: f j 2-- Kris Hanson, Ph.D. 3M Study Director Date IZ' >CT.`7<? Date f t QJr f f Date 3M Environmental Laboratory ADVANCED BIOANALYTICAL S E R V IC E S , INC. Page 2 of3 99ADJA02.MI.ADD.DOC Page 247 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 ADDENDUM SUMMARY ADDENDUM Addition of Study Director signature on the Signature Page. The signature page of this addendum indicates acceptance of the original report. REASON FOR ADDENDUM Original GLP report requires the signature of the Study Director. <***, ADVANCED BIOANALYTICAL S E R V IC E S , INC. 3M Environmental Laboratory Page 3 of 3 99ADJA02.MI.ADD.DOC Page 248 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 RE-ISSUED ANALYTICAL REPORT STUDY TITLE Oral (Gavage) Pharmacokinetic Study of PFOS in Rats DATA REQUIREMENTS Analytical Method Requirements STUDY DIRECTOR Marvin T. Case, 3M PRINCIPAL INVESTIGATOR Enaksha Wickremesinhe, Centre RE-ISSUED ANALYTICAL REPORT COMPLETION DATE October 19,2000 PERFORMING LABORATORY / TESTING FACILITY Centre Analytical Laboratories, Inc. (Centre) 3048 Research Drive State College, PA 16801 Phone: 814-231-8032 STUDY SPONSOR 3M Toxicology Services - Medical Department 3M Center, Building 220-2E-02 St. Paul, MN 55144-1000 PROJECT IDENTIFICATION Sponsor Protocol Number: FACT-TOX-110 Centre Study Number: 023-016 Total Pages: 103 3IVI Environmental Laboratory Page 249 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT Centre Study Number 023-016, entitled "Oral (Gavage) Pharmacokinetic Study of PFOS in Rats," conducted for 3M Environmental Toxicology Services - Medical Department, was performed in compliance with US FDA Good Laboratory Practice Standards (21 CFR 58) by Centre Analytical Laboratories, Inc. with the following exceptions: 1. The reference substances (analyticd standards) used in this study (PFOS and THPFOS) have not been characterized under 21 CFR 58.105 2. The automated data collection systems used in this study are not fully compliant with 21 CFR 58.130(e). /\ ft . / A '-- - FOA- eN A fkJM w t/l& n & fr tih ? Enaksha Wickremesinhe, Ph.D* Principal Investigator Centre Analytical Laboratories, Inc. /f-cr-a Date *No longer employed at Centre. However, facility management, Rick Grazzini will be signing in the principal investigator's stead. Marvin T Case, Ph.D. Study Director 3M Toxicology Services Date Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 2 of 103 Page 250 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 QUALITY ASSURANCE STATEMENT Centre Analytical Laboratories' Quality Assurance Unit reviewed Centre Study Number 023-016, entitled, "Oral (Gavage) Pharmacokinetic Study of PFOS in Rats". All phases were reviewed for conduct according to Centre Analytical Laboratories' Standard Operating Procedures, the Study Protocol, and all applicable Good Laboratory Practice Standards. All findings were reported to the Study Director and to management. Phase 1. Protocol Review Date InsDected 5/17/00 Date Reported to Date Reported to Centre Study Director and Management SDonsor Management 6/1/00 7/14/00 2. Extraction 6/27/00 7/11/00 7/14/00 3. Raw Data Review 7/10,13,14,20/00 7/29/00 8/25/00 4. Draft Report Review 8/3-4/00 8/15/00 8/25/00 5. Final Report Review 9/8/00 9/11/00 9/11/00 ____________ <xJIjlqa*-- Naomi Lovallo Quality Assurance Auditor _____ (q{/ 9/oe Date Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 3 of 103 Page 251 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 CERTIFICATION OF AUTHENTICITY This report, for Centre Study Number 023-016, is; a true and complete representation of the raw data for the study. Submitted by: Centre Analytical Laboratories, Inc. 3048 Research Drive State College, PA 16801 (814)231-8032 Principal Investigator, Centre: f - 0(X' to Enaksha Wickremesinhe, Ph.D.* Group/Team Leader Centre Analytical Laboratories, Inc. Date *No longer employed at Centre. However, facility management, Rick Grazzini will be signing in the principal investigator's stead. Centre Analytical Laboratories, Inc. Facility Management: John Flaherty / Laboratory Manager Centre Analytical Laboratories, Inc. Date Study Director, 3M: TC-JJlJL,_________ 23 Marvin T. Case, Ph.D. 3M Toxicology Services Date Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 4 of 103 Page 252 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 STUDY IDENTIFICATION Oral (Gavage) Pharmacokinetic Study of PFOS in Rats SPONSOR PROTOCOL NUMBER: FACT-TOX-110 TYPE OF STUDY: Oral (Gavage) Pharmacokinetic TEST SYSTEM: Rat Feces TEST MATERIAL: SPONSOR: STUDY DIRECTOR: Perfluorooctiine sulfonic acid potassium salt (T-6295) 3M Toxicology Services - Medical Department 3M Center, Building 220-2E-02 St. Paul, MN 55144-1000 From 6/10/99 to 2/10/00: Kris Hansen, PhD. 3M Environmental Technology and Safety Services Phone:(651)778-6018 From 2/10/00 to Present: Marvin T. Case, D.V.M., Ph.D. 3M Toxicology Services Phone:(651)733-5180 TESTING FACILITY: Centre Analytical Laboratories, Inc. 3048 Research Drive State College, PA 16801 PRINCIPAL INVESTIGATOR Enaksha Wickremesinhe, Ph.D. Centre Analytical Laboratories, Inc. Phone:(814)231-8032 ANALYTICAL PHASE TIMETABLE: Study Initiation Date: Analytical Start Date: Analytical Termination Date: 06/10/99 06/09/00 07/01/00 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Pag 5 o f 103 Page 253 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 PROJECT PERSONNEL The Study Director for this project was Marvin T. Case at 3M Toxicology Services and the Principal Investigator for this project at Centre Analytical Laboratories, Inc. was Enaksha Wickremesinhe. The following personnel from Centre Analytical Laboratories, Inc., were associated with various analytical phaseis of the study: Name . Enaksha Wickremesinhe Emily Stauffer . Karen Smith Tiffany Proctor Rickey Keller Lawrence Ord ' Title Group/Team Leader Scientist Scientist Technician Simple Custodian S;imple Custodian Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 6 of 103 Page 254 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 TABLE OF CONTENTS Page TTTLE PAGE...................................... 1 GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT.............................. 2 QUALITY ASSURANCE STATEMENT.................................................................... 3 CERTIFICATION OF AUTHENTICITY..................................... ;.................................... 4 STUDY IDENTIFICATION................................................................................................5 PROJECT PERSONNEL.............................................................................................. 6 TABLE OF CONTENTS.....................................................................................................7 LIST OF TABLES .............................................. ^.............................................................. 8 LIST OF FIGURES......... .........................................................................................-,........9 LIST OF APPENDICES................................................................ 10 1.0 SUMMARY.............................................................................. 11 2.0 OBJECTIVE....... '........................................................................................................ 11 3.0 INTRODUCTION....................................................................................................... 11 4.0 TEST SYSTEM..........................................................................................................U 5.0 REFERENCE MATERIAL......................................................... 12 6.0 EXPERIMENTAL DESIGN....................................................................................... 13 7.0 DESCRIPTION OF ANALYTICAL METHOD........................................................ 13 7.1 Extraction Procedure............................................................................................ 13 7.2 Preparation of Standards and Fortification Solutions........................................... 13 7.3 Chromatography....................................... 14 7.4 Instrument Sensitivity............................... 14 . 7.5 Description of Instrument and Operating Conditions............................................14 7.6 Quantitation and Example Calculation..................................................................16 8.0 RESULTS AND DISCUSSION................................................................................18 9.0 CIRCUMSTANCES THAT MAY HAVE AFFECTED THE DATA..................... 18 10.0 RETENTION OF DATA AND SAMPLES...............................................................18 11.0 TABLES............................................... 19 12.0 FIGURES............................................................:................................................... 40 13.0 APPENDICES............................................................................... 48 Centre Analytical Laboratories, Inc. 3IVI Environmental Laboratory Page 7 of 103 Page 255 _ 3M Medical Department Study: T-629S.12 . Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 LIST OF TABLES Page Table I. Summary of PFOS residues in Matrix Blanks and Matrix Zero Blanks............ 20 Table n. Summary of PFOS residues in Gl-Control-GDO..............................................22 Table IB. Summary of PFOS residues in G2-0.1MKD On GD0......................................23 Table IV. Summary of PFOS residues in G3-0.4MKD on GD0 ..................................24 Table V. Summary of PFOS residues in G4-1.6MKD on GD0....................................... 25 Table VI. Summary of PFOS residues in G5-3.2MKD on GD0......................................26 Table VII. Summary of PFOS residues in Gl-Control on GD7....................................... 27 Table VUL Summary of PFOS residues in G2-0.1MKD on G D 7............... ................... 28 Table IX. Summary of PFOS residues in G3-0.4MKD on GD7......................................29 Table X. Summary of PFOS residues in G4-1.6MKD on GD7....................................... 30 Table XL Summary of PFOS residues in G5-3.2MKD on GD7......................................31 Table XIL Summary of PFOS residues in Gl-Control on GD15..................................... 32 Table Xm. Summary of PFOS residues in G2-0.1MKD on GD15.............. :................. 33 Table XIV. Summary of PFOS residues in G3-0.4MKD on GDI 5 ................................. 34 Table XV. Summary of PFOS residues in G4-1.6MKD on GD15..................................35 Table XVI. Summary of PFOS residues in G5-3.2MKD on GD15................................. 36 Table XVII. Summary of PFOS residues in Gl-Control on GD21.................................. 37 Table XVm. Summary of PFOS residues in G2-0.1MKD on GD 21.......... ................... 37 Table XIX. Summary of PFOS residues in G3-0.4MKD on GD21................................. 38 Table X X Summary of PFOS residues in G4-1.6MKD on GD21.................................. 38 Table XXI. Summary of PFOS residues in G5-3.2MKD on GD21................................. 38 Table XXH. Summary of PFOS recoveries in Fortified Samples........................ ............ 39 ' Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 8 of 103 Page 256 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 . Sponsor Protocol No: FACT-TOX-110 LIST OF FIGURES .- Page Figure 1. Typical Calibration Curve for PFOS................................................................41 Figure 2. Typical Mean Response Factor for THPFOS..................................................4 2 Figure 3. Chromatogram Representing a 5 ng/ml, extracted standard for PFOS and 250 ng/mL fortification of THPFOS.....................................................;...............43 Figure 4. Chromatogram Representing a 125 ng/mL extracted standard for PFOS and 250 ng/mL fortification of THPFOS............................................................... 44 Figure 5. Chromatogram Representing Control Rat Feces for PFOS and THPFOS (Centre ID: 0004277 Blank B, Set: 062300A)............. .................................45 Figure 6. Chromatogram Representing Control Rat Feces Fortified with 50 ng of PFOS and 500 ng of THPFOS (Centre ID: 0003981 Spk A, Set: 061400A)........ 46 Figure 7. Chromatogram of Rat Feces Sample from G2 on GD0 (Centre ID: 0003994, Set: 061400A).................................................................................................47 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 9 of 103 Page 257 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-1 10 LIST OF APPENDICES Page Appendix A Study Protocol FACT-TOX-110 (Centre Study No. 023-016) and Amendments and Deviations .......... 49 Appendix B Determination of Fluorochemical Residues in Monkey/Rat Feces by LC/MS/MS (Revision 2) Method #COM-023-003, revision 2 and Deviations and Modifications......................... 75 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 10 of 103 Page 258 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 1.0 SUMMARY The purpose of this study was to analyze residues of perfluorooctanesulfonate (PFOS) in rat feces as specified in 3M Protocol FACT-TOX-110. The analytical method used for this study was entitled, "Determination of Fluorochemical Residues in Monkey/Rat Feces by LC/MS/MS, revision 2" (Centre method number: 00M-023-003, revision 2). The limit of quantification for PFOS in rat feces was 10 ppb. Residues (corrected for purity) ranging from non-detected levels to 59386.8 ppb were found in the rat feces samples. Fortification recoveries ranged from 71-128% with an average of 98% and relative standard deviation of 15%. 2.0 OBJECTIVE The objective of this study was to determine levels of perfluorooctanesulfonate (PFOS) in specimens of feces of rats using the analytical method entitled "Determination of Fluorochemical Residues in Monkey/Rat Feces by LC/MS/MS, revision 2." 3.0 INTRODUCTION This report details die results of the residues of PFOS detected in rat feces, using the analytical method entitled, "Determination of Fluorochemical Residues in Monkey/Rat Feces by LC/MS/MS (revision 2).'' Complete detedls of the analytical methodology can be found in Appendix B. The study was initiated on June 10, 2000, when the study director signed the protocol FACT-TOX-110 Amendment #4. The complete protocol and amendments can be found in Appendix A. The analytical start date was June 9,2000, and the analytical termination date was July 1,2000. 4.0 TEST SYSTEM The 250 rat feces samples analyzed in this study were received frozen on dry ice from 3M Environmental Laboratory St. Paul, MN on May 26, 2000 and stored frozen upon receipt. The samples were then logged in on May 31,2000 by Centre personnel and transferred to different frozen storage locale. . The control rat feces used for standards and blanks was purchased from Lampire Biological Laboratories, Inc., Pipersville, PA and received at Centre frozen on dry ice on Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 11 of 103 Page 259 31V! Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 May 4, 2000 and June 22, 2000, logged in by Centre personnel and placed in frozen storage (<-10C). Sample login and chain of custody information can be found in the raw data package associated with this study. Storage records will be kept at Centre Analytical Laboratories, Inc. and a true copy of the storage records can be found in the raw data package associated with this study. 5.0 REFERENCE MATERIAL The analytical standard PFOS (as the potassium salt) was received at Centre on November 12, 1999 and the surrogate standard THPFOS was received on December 3, 1999 from 3M Environmental Technology and Services. Characterization of the reference material . PFOS was performed on August 31, . 2000 at Centre and documentation can be found in raw data package associated with this report. The available information for the reference materials is listed below. The reference materials were stored at room temperature. Compound PFOS THPFOS Centre Control No. 99-023-002 99-023-011 Batch No. 217 53406 Purity (%') Expiration Date 86.9 08/31/01 NA 01/01/10 Molecular structures of PFOS and THPFOS are given below. PFOS Chemical Name: Perfluorooctane sulfonate Molecular weight: 499 (C8F17SO3-) o C gF ^S CT |. Note: The neutral molecule and standard form from which PFOS (anion) is derived, is potassium perfluorooctane sulfonate [CgFnSOjK], molecular weight 538. THPFOS Chemical Name: 4-H, perfluorooctane sulfonic acid Molecular weight: 428 . 1-H, 1-H, 2-H, 2-H, C8FI3S03H Centre Analytical Laboratories, Inc. 3IVI Environmental Laboratory Page 12 of 103 Page 260 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 : 6.0 EXPERIMENTAL DESIGN Samples were extracted according to group number and sampling day. Each set of samples contained two matrix blanks, two matrix blanks spiked- with the surrogate standard (zero blank), two matrix spikes, and 1 2-16 samples. The extracts were analyzed by LC/MS/MS. Samples with residues outside the linear range of the calibration curve were diluted and re-analyzed. 7.0 DESCRIPTION OF ANALYTICAL METHOD Analytical method entitled "Determination of Fluorochemical Residues in Monkey/Rat Feces by LC/MS/MS (Revision 2)" was used for this study. 7.1 Extraction Procedure One gram 0.05 of sample was weighed into 20 mL polypropylene scintillation vials and fortified (if necessary) using disposable micropipettes. Ten mL of acetonitrile was added to the vial, capped tightly, and placed on a wrist-action shaker for ~ 30 min. The samples were filtered through a glass acrodisc filter. The filtered extract was passed through a conditioned carbon SPE column and collected. The columns were then eluted with -10 mL acetonitrile followed by -20 mL 90:10 acetonitrile:2% ascorbic acid in methanol. The combined extracts were evaporated down to almost dryness with a rotary evaporator and then re-constituted with methanol, making final volume 2 mL. The .samples were analyzed using electrospray LC/MS/MS. 7.2 Preparation of Standards and Fortification Solutions Standard solutions were prepared on May 3, 2000 as specified in Centre Analytical Laboratories' analytical method entitled, "Determination of Fluorochemical Residues in Monkey/Rat Feces by LC/MS/MS (Revision 2)." An individual stock standard solution of PFOS was prepared at a concentration of 100 pg/mL by dissolving 10 mg of the standard (corrected for salt purity only) in methanol. From this solution, a 10 pg/mL fortification standard solution was prepared by taking 10 mL of the stock and bringing the volume up to 100 mL with methanol. Also, a stock standard of THPFOS was prepared at 100 pg/mL by dissolving 10 mg of the standard in methanol. An individual 10 pg/mL solution of THPFOS was prepared in the same fashion described above. A 2.5 pg/mL fortification standard of PFOS was prepared by taking 25 mL of the 10 pg/mL solution of PFOS and bringing the volume up to 100 mL with methanol. An individual fortification solution of THPFOS was prepared in the same manner. A 0.5 pg/mL PFOS standard was prepared by taking 20 mL of the 2.5 pg/mL PFOS Centre Analytical Laboratories, Inc. Page 13 of 103 ; ! j j \ j ' j l j ; ' : . 3M Environmental Laboratory Page 261 3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 Centre Study No.: 023-016 ' Sponsor Protocol No: FACT-TOX-110 standard and bringing to 100 niL with methanol. To make the 0.1 pg/mL PFOS fortification standard, 20 mL of the 0.5 pg/mL of PFOS was brought to 100 mL with methanol. Calibration standards were processed through the extraction, procedure, identical to the samples. The fortification of the standards before extraction was done according to the following table: - Cone, of PFOS Fortification Fortification Volume (jiL) Solution (pg/ml) Weight of Control Simple (g) 0.05 0.1 100 1.0 0.1 200 1.0 0.5 100 1.0 0.5 200 1.0 2.5 100 1.0 2.5 200 1.0 * 2.5 pg/ml THPFOS fortification solution. Fon. Level of Voi. of Extracted Surrogate Calibration Standard* Standard added(pL) (pDb) 10 200 20 200 50 200 100 200 250 200 500 200 The stock standard solution and all fortification and calibration standard solutions were stored in a refrigerator (4 2C) when not in use. Documentation of standard preparation and extraction can be found in the raw data associated with this report. 13Chromatography ' Quantification of PFOS and THPFOS was accomplished by LC/MS/MS analysis using electrospray LC/MS/MS. The retention times of PFOS and THPFOS were - 5.2 min. and - 5.0 min., respectively, with no significant interfering peaks in the control matrices corresponding to either of the analyte retention times. 7.4 Instrum ent Sensitivity The smallest standard amount injected during the chromatographic run was equivalent to 5 ng/mL of PFOS and 250 ng/mL of THPFOS in the rat feces matrix. 7.5 Description of Instrum ent and Operating Conditions A Micromass Quattro Ultima LC/MS/MS coupled to a Hewlet Packard HPLC system was used. Data acquisition and processing were performed using Masslynx 3.4 software. Detailed operating conditions are listed below: Centre Analytical Laboratories, Inc. Page 14 of 103 3M Environmental Laboratory Page 262 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-VIO LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Instrument: Micromass Quattro Ultima ELECTROSPRAY ION SOURCE; Capillary: 3.0 kV Hexapole 2: 0.3 V Hexapole 1:0.1 V Aperture 1:0.2 V Source Block Temp.: 100C Desolvation Temp.: 350C ANALYZER: LMRes 1 :10.5 V HM Res 1:10.5 V Energy 1:1.0 V Entrance: -2 V Exit: 2 V LM Res 2: 13.0 V HM Res 2: 13.0 V IEnergy 2: 2.0 V Multiplier: 650 V GAS FLOWS AND PRESSURE: Desolvation N2Flow Rate: -650 L/hr Nebuliser N2Flow Rate: -150 L/hr Gas Cell Pressure: -0.003 mbar Computer COMPAQ Professional Workstation AP200 Software: Microsoft WindowsNT: Version4BuiId 1381: Service Pack5 Micromass Limited: Masslynx 3.4 Build 004 HPLC Equipment: Hewlett Packard (HP) Series 1100 HPBinary Pump " HP Vacuum Degasser HP Autosampler HP Column Oven HPLC Column: Genesis C-8,5 cm x 2.1 mm i.d. x 4 p Column Temperature: 35C ' Mobile Phase (A) : 2 mM AmmoniumAcetate in Type I Water Mobile Phase (B) : Methanol Gradient: Injected Volume: Ions monitored : Analyte PFOS THPFOS Time (min) 0.0 0.4 1.0 7.0 7.5 10.5 11.0 14.5 15.0 10 pL %A 60.0 60.0 10.0 10.0 0.0 0.0 60.0 60.0 60.0 Transition M onito red 499 -* 99 427 SO Dwell (secs) 0.2 0.2 %B 40.0 40.0 90.0 90.0 100.0 100.0 40.0 40.0 40.0 Flow Rate (mL/min) 0.3 0.3 0.3 0.3 0.3 0.4 0.4 0.4 0.3 Coll Enersv (eV) Cone (V) 43 76 35 34 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 15 of. 103 Page 263 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 7.6 Quantitation and Example Calculation Centre Study No.: 023-016 . Sponsor Protocol No: FACT-TOX-110 Ten microliters of sample or extracted calibration standard was injected into the LC/MS/MS. The peak area was measured and the standard curve was generated (using 1/x weighted linear regression) by Masslynx software using six concentrations of standards. The surrogate standard, THPFOS, was only used to monitor the efficiency of the extraction procedure and was not used for quantitation of PFOS. The residue concentration in rat feces was determined using the; following equations: Equations 1 and 2 were used to calculate the amount of analyte found (in ppb, based on peak area) using the standard curve generated by the Masslynx software program. Equation 1: Analyte found (ng/mL) = (Peak area - intercept! slope For samples fortified with known amounts of anal;rtes prior to extraction, use Equation 3 to calculate the percent recovery. Equation 3: Recovery (%) = [analytefound (ng/mL)xfinal vol. (:mL)xDF] analyte added (ng) Equation 4: This calculation wasonlyperformedonactual ssunplesandnot QCrecoveries. Corrected analyte found (ppb) = analyte found (ppb) x % purity % purity for PFOS lot 217 = 86.9% (0.869) An example of a calculation using an actual sample analyzed with extracted standards follows: Rat feces sample Centre ID 0004127 Spk A (Set: 062700A), fortified at 100 ng with PFOS. Where: peak area intercept slope 32320 2653.87 595.935 dilution factor 1 Centre Analytical Laboratories, Inc. Page 16 of 103 3IVI Environmental Laboratory Page 264 3M Medical Department Study: T-6295.12 : , ' Analytical Report: FACT TOX-110 LRN-U2849 ng added (fort level) final voi. sample wt. = = = Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 100 2mL 1.01 g From equation 1: Analyte found (ng/mL) . = r32320- 2653.871 595.935 = 49.8 ng/mL From equation 2: Analyte found (ppb) = i4 9 .8 x 2 m L x 11 1.01 g = 98.6 ppb From equation 3: % Recovery = 149.8 ng/mL x 2 m l x 11 x 100 100 ng = 100% Note: This example calculation was done using rounded numbers, and therefore may be slightly different from the values shown in the RAW DATA. The amount of surrogate standard THPFOS found was calculated using the following equation: ' Analyte found (ng/mL): peak area mean response factor Other statistical methods used in analyzing this data. were: StandardDeviation= i ito- * ) 8 h rr- Mean= x= n 1 RelativeStandardDeviation(RSDorCoefficientofVariation(CV))= Standard Deviation x 100% Mean Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 17 of 103 Page 265 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 8.0 RESULTS AND DISCUSSION Although, the majority of the control samples did not contain significant interferences, a couple of samples did contain residues that were comparable to control group levels. A summary of residues found in all of the matrix blanks and matrix zero blanks is detailed in Table I. Residues (corrected for purity) ranging from non-detected levels to 59386.8 ppb were found in the rat feces samples. The residues found in all of the samples plus the averages and standard deviations for each group at each interval are detailed in Tables II-XX I. For this report the residues found in Tables I-XXI were corrected for the purity of the PFOS standard lot 217 based on the certificate of analysis finalized on September 7, 2000. Fortification recoveries ranged from 71-128% with an average of 98% and relative standard deviation of 15% (n = 34). A summary of all of the fortification recoveries can be found in Table XXII. Typical calibration curves and chromatograms representing standards, controls, fortifications, and samples are depicted in Figures 1-7. 9.0 CIRCUMSTANCES THAT MAY HAVE AFFECTED THE DATA Electronic records are not fully compliant with 21 CFR 11, "Electronic Records: Electronic Signature." However, fully approved SOP's were in' place and all chromatography instrumentation used in this study was fully validated (IQ, PQ, OQ), calibrated and operational. All original data were printed as hard copies and fiilly audited by quality assurance. Verified exact copies and the electronic data have been stored in the archives at Centre Analytical Laboratories, Inc. Original raw data have been returned to the sponsor. 10.0 RETENTION OF DATA AND SAMPLES When the final report is complete, all original paper data generated by Centre Analytical Laboratories, Inc. will be shipped to the sponsor. This does not include facility-specific raw data such as instrument logs, however exact copies of temperature logs will be submitted. Exact copies of all raw data, as well as a signed copy of the final analytical report and all original facility-specific raw data, will be retained in the Centre Analytical Laboratories, Inc. archives for the period of time sjjecified in 21 CFR Part 58. Retained samples of reference substances are archived by the sponsor. Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 18 of 103 Page 266 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 11.0 TABLES Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 19 of 103 Page 267 3M Medical Department Study: T-6295.12 , Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Table I. Summary of PFOS residues in Matrix Blanks and Matrix Zero Blanks ND = Not Detected and NQ = Not Quantifiable Sponsor Centre Residue C u n icfed Set Date Found Residue Found ID ID Number Extracted (ng/g) Cng/g) na 0004277 Blank A 061200AR 6/12/00 : ND na 0004277 Blank B 061200AR 6/12/00 ND ND ND na 0004277 Zero Blank C 061200AR 6/12/00 ND ND na 0004277 Zero Blank D 061200AR 6/12/00 ND ND na 0004277 Blank A 061400A 6/14/00 NQ NQ na 0004277 Blank B 061400A 6/14/00 NQ NQ na 0004277 Zero Blank C 061400A 6/14/00 NQ NQ na 0004277 Zero Blank D 06140QA 6/14/00 ND ND na 0004277 Blank A 061500AD 6/15/00 NQ NQ na 0004277 Blank B 061500AD 6/15/00 NQ NQ na 0004277 Zero Blank C 061500AD 6/15/00 1.6 1.4 na 0004277 Zero Blank D 061500AD 6/15/00 1.3 1.1 na 0004277 Blank E 061600A 6/16/00 2.0 1-7 . na 0004277 Blank F 061600A 6/16/00 NQ NQ na 0004277 Zero Blank G ' 061600A 6/16/00 NQ NQ na 0004277 Zero Blank H 061600A 6/16/00 NQ NQ na 0004277 Blank A 061900AD 6/19/00 NQ NQ na 0004277 Blank B 061900AD 6/19/00 NQ NQ na 0004277 Zero Blank C 061900AD 6/19/00 NQ NQ na 0004277 Zero Blank D 061900AD 6/19/00 NQ NQ na 0003466 Blank A 062000A 6/20/00 NQ NQ na 0003466 Blank B 062000A 6/20/00 NQ NQ n 0003466 Zero B lank C 062000A 6/20/00 NQ NQ na 0003466 Zero Blank D 062000A 6/20/00 NQ NQ na 0004277 Blank A 062100A 6/21/00 NQ NQ na 0004277 Blank B 062100A 6/21/00 NQ NQ na 0004277 Zero Blank C 062100A 6/21/00 NQ NQ na 0004277 Zero Blank D 062100A 6/21/00 NQ NQ na 0004277 Blank A 062200A 6/22/00 6.5 5.6 na 0004277 Blank B 062200A 6/22/00 ND ND na 0004277 Zero Blank C 062200A 6/22/00 ND ND na 0004277 Zero Blank D 062200A 6/22/00 ND ND na 0004277 Blank E 062200B 6/22/00 ND ND na 0004277 Blank F 062200B 6/22/00 ND ND na 0004277 Zero Blank G 062200B 6/22/00 ND ND Centre Analytical Laboratories, Inc. Page 20 o f 103 3M Environmental Laboratory Page 268 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 fable I (cont.) Summary of PFOS residues in Matrix Blanks and latrix Zero Blanks ND = Not Detected and NQ = Not Quantifiable Sponsor Centre ID Residue Corrected Residue Set Date Found Found ID Number Extracted fog/g) (ng/g) na 0004277 Zero Blank H 062200B 6/22/00 ND ND na 0004277 Blank A 062300A 6/23/00 ND : ND na 0004277 Blank B 062300A t m m ND ND na 0004277 Zero Blank C 062300A 6/23/00 ND ND na 0004277 Zero Blank D 062300A 6/23/00 ND ND aa 0004277 Blank A 062300B 6/23/00 NQ NQ la 0004277 Blank B 062300B 6/23/00 NQ NQ ia 0004277 Zero Blank C 062300B 6/23/00 NQ NQ \a 0004277 Zero Blank D 062300B 6/23/00 NQ NQ a 0005738 Blank A 062600A 6/26/00 0.5 0.4 a 0005738 Blank B 062600A 6/26/00 12.6 10.9 na 0005738 Zero Blank C 062600A 6/26/00 ND ND na 0005738 Zero Blank D 062600A 6/26/00 ND ND na 0005738 Blank E 062600B 6/26/00 ND ND na 0005738 Blank F 062600B 6/26/00 ND ND na 0005738 Zero Blank G 062600B 6/26/00 ND ' ND na 0005738 Zero Blank H 062600B 6/26/00 ND ND na 0005738 Blank A 062700A 6/27/00 ND ND na 0005738 Blank B 062700A 6/27/00 ND ND na 0005738 Zero Blank C 062700A 6/27/00 ND ND na 0005738 Zero Blank D 062700A 6/27/00 ND . ND na 0005738 Blank A 062800A 6/28/00 ND ND na 0005738 Blank B 062800A 6/28/00 ND na 0005738 Zero Blank C 062800A 6/28/00 ND ND ND na 0005738 Zero Blank D 062800A 6/28/00 ND ND na 0005738 Blank A 062900A 6/29/00 30.1 26.2 na 0005738 Blank B 062900A 6/29/00 ND ND na 0005738 Zero Blank C 062900A 6/29/00 ND ND na 0005738 Zero Blank D 062900A 6/29/00 ND ND na 0005738 Blank A 063000A 6/30/00 ND ND na 0005738 Blank B 063000A 6/30/00 ND ND na 0005738 Zero Blank C 063000A 6/30/00 ND ND na 0005738 Zero Blank D 063000A 6/30/00 ND AVERAGE: 0.8 ND 0.7 STANDARD DEVIATION: 4 3 Centre Analytical Laboratories, Inc. Page 21 of 103 3M Environmental Laboratory Page 269 31V Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Table n . Summary of PFOS residues in Gl-Control-GDO Sponsor ID 19501F.F0,Gl-Control-GDO 19502FJ0,Gl-Control-GDO 19503FJF0,Gl-Control-GDO 19504FJO,Gl-Control-GDO 19505FJO,Gl-Control-GDO 19506FJ0,Gl-Control-GD0 19507FJ0,Gl-Control-GDO 19508FJO,Gl-Control-GDO 19509FJO,Gl-Control-GDO 19510FJO,Gl-Control-GDO 1951 IF ,F0,G1 -Control-GDO 19512FJO,Gl-Control-GDO 19513FJO,Gl-Control-GDO 19514F,F0,G1-Control-GDO 19515FJO,Gl-Control-GDO 19516FJ0.G1 -Control-GDO Centre ID Set Corrected Date Residue Found Residue Found Number Extracted (ns/g) (nst/g) 0003977 061200AR 6/12/00 0.0 0.0 0003978 061200AR 6/12/00 0.0 0.0 : 0003979 061200AR 6/12/00 5.9 5.1 0003980 061200AR 6/12/00 0.0 0.0 0003981 061200AR 6/12/00 0.0 0.0 0003982 061200AR 6/12/00 30.1 26.2 0003983 061200AR 6/12/00 0.0 0.0 0003984 061200AR 6/12/00 5.9 5.1 0003985 061200AR 6/12/00 40.0 34.8 0003986 061200AR 6/12/00 0.0 0.0 0003987 061200AR 6/12/00 0.0 0.0 0003988 061200AR 6/12/00 0.0 0.0 0003989 061200AR 6/12/00 0.0 0.0 0003990 061200AR 6/12/00 0003991 061200AR 6/12/00 0.0 0.0 0.0 0.0 0003992 061200AR 6/12/00 '60.9 52.9 AVERAGE: 8.9 7.8 STANDARD DEVIATION: 18 16 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 22 of 103 Page 270 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 , Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Table III. Summary of PFOS residues in G2-0.1MKD on GDO Sponsor Centre ID Set Corrected Date Residue Found Residue Found ID Number Extracted (na/g) (na/g) 19517F.F0.G2-0.1MKD-GD0 0003993 061400AD 6/14/00 700.8 609.0 19518F.F0.G2-0.1MKD-GD0 0003994 061400A 6/14/00 455.7 396.0 19519FJ0.G2-0.1MKD-GD0 0003995 061400A 6/14/00 389.7 338.6 19520FJ0.G2-0.1MKD-GD0 0003996 061400AD 6/14/00 675.5 587.0 19521F.F0.G2-0.1MKD-GD0 0003997 061400A 6/14/00 494.8 430.0 19522F.F0.G2-0.1MKD-GD0 0003998 061400A 6/14/00. 381.0 331.1 19523F.F0.G2-0.1MKD-GD0 0003999 061400A 6/14/00 458.2 398.2 19524FJF0.G2-0.1MKD-GD0 0004000 061400AD 6/14/00 832.8 723.7 19525FvF0,G2-0.1MKD-GD0 0004001 061400A 6/14/00 473.0 411.0 19526F,F0,G2-0.1MKD-GD0 0004002 061400A 6/14/00 446.1 387.7 19527F,F0,G2-0.1MKD-GD0 0004003 061400A 6/14/00 497.4 432.2 19528FJF0.G2-0.1MKD-GD0 0004004 061400AD 6/14/00 700.7 608.9 19529FJ0.G2-0.1MKD-GD0 0004005 061400AD 6/14/00 796.9 692.5 19530FF0.G2-0.1MKD-GD0 0004006 061400A 6/14/00 393.7 342.1 19531F,F0,G2-0.1MKD-GD0 0004007 061400AD 6/14/00 734.7 -638.5 19532F,F0,G2-0.1MKD-GD0 0004008 061400AD 6/14/00 748.2 650.2 AVERAGE: 573.7 498.5 STANDARD DEVIATION: 160 139 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 23 of 103 Page 271 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 . Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Table IV. Summary of PFOS residues in G3-0.4MKD on GDO Sponsor Centre ID Set ID Number 19533F,F0,G3-0.4MKD-GD0 0004009 061500AD Corrected Date Residue Found Residue Found Extracted (na/g) (ng/g) 6/15/00 2598.1 2257.7 19534FJF0.G3-0.4MKD-GD0 0004010 061500AD 6/15/00 19535F,F0,G3-0.4MKD-GD0 0004011 061500AD 6/15/00 3750.7 2801.3 3259.4 2434.3 19536F,F0,G3-0.4MKD-GD0 0004012 061500AD 19537FJ0.G3-0.4MKD-GD0 0004013 061500AD 19538FJ0.G3-0.4MKD-GD0 0004014 061500AD 6/15/00 6/15/00 6/15/00 3174.3 2777.9 3083.8 2758.5 2414.0 2679.8 19539FJ0.G3-0.4MKD-GD0 0004015 061500AD 6/15/00 19540F.F0.G3-0.4MKD-GD0 0004016 061500AD 6/15/00 3680.4 2044.0 3198.3 ' 1776.2 19541F,F0,G3-0.4MKD-GD0 0004017 061500AD 6/15/00 2104.0 1828.4 19542F,F0,G3-0.4MKD-GD0 0004018 061500AD 6/15/00 3290.2 2859.2 19543F.F0.G3-0.4MKD-GD0 0004019 061500AD 6/15/00 19544FJF0.G3-0.4MKD-GD0 0004020 061500AD 6/15/00 2113.4 3430.8 1836.5 2981.4 19545FJ0.G3-0.4MKD-GD0 0004021 061600AD 6/16/00 19546F.F0.G3-0.4MKD-GD0 0004022 061500AD 6/15/00 2755.3 2334.2 2394.4 2028.4 19547F,F0,G3-0.4MKD-GD0 0004023 061500AD 6/15/00 2215.6 1925.4 19548F.FO.G3-O.4MKD-GD0 0004024 061500AD 6/15/00 2334.8 2028.9 AVERAGE: 2780.6 24163 STANDARD DEVIATION: 570 496 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 24 of 103 Page 272 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 . Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Table V. Summary of PFOS residues in G4-1.6MKD on GDO Sponsor Centre ID Set Corrected Date Residue Found Residue Found ID Number Extracted (ne/g) (ng/g) 19549F,F0,G4-1.6MKD-GD0 0004025 061600AD 6/16/00 19550FJF0.G4-1.6MKD-GD0 0004026 061600AD 6/16/00 14122.8 16858.7 12272.7 14650.2 19551F,F0,G4-1.6MKD-GD0 0004027 061600AI) 6/16/00 10394.9 9033.2 19552F,F0,G4-1.6MKD-GD0 0004028 061600AD 6/16/00 13143.8 11422.0 19553FJ0.G4-1.6MKD-GD0 0004029 061600AD 6/16/00 13712.0 11915.7 19554F,F0,G4-1.6MKD-GD0 0004030 061600D 6/16/00 15507.9 13476.4 19555FJ0.G4-1.6MKD-GD0 0004031 061600AD 6/16/00 10328.8 8975.7 19556FJ0,G4-1.6MKD-GD0 0004032 061600AD 6/16/00 10253.1 8909.9 19557FJF0.G4-1.6MKD-GD0 0004033 061600AD 6/16/00 8961.3 7787.4 19558F.F0.G4-1.6MKD-GD0 0004034 061600AD 6/16/00 6845.2 5948.5 19559FJF0.G4-1.6MKD-GD0 0004035 061600AD 6/16/00 7677.3 6671.6 19560FJ0,G4-1.6MKD-GD0 0004036 061600AD 6/16/00 16518.0 14354.1 19561FJ0.G4-1.6MKD-GD0 0004037 061600AD 6/16/00 12121.6 10533.7 19562F,F0,G4-1.6MKD-GD0 0004038 061600AD 6/16/00 9585.4 8329.7 19563F,F0,G4-1.6MKD-GD0 0004039 061600AD 6/16/00 7465.7 6487.7 19564FJF0.G4-1.6MKD-GD0 0004040 061600AD1 6/16/00 16957.0 14735.6 AVERAGE: 119033 10344.0 STANDARD DEVIATION: 3453 3000 Centre Analytical Laboratories, Inc. 3Mi Environmental Laboratory Page 25 of 103 Page 273 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 . Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Table VI. Summary of PFOS residues int G5-3.2MKD on GDO Sponsor Centre ID Set Corrected Date Residue Found Residue Found ID Number Extracted (ns/.s) u.a/g) 19565F,FO,G5-3.2MKD-GDO 0004041 061900AD 6/19/00 23450.7 20378.7 19566FF0,G5-3.2MKD-GD0 0004042 061900AD 6/19/00 29548.0 25677.2 19567F,FO,G5-3.2MKD-GDO 0004043 061900AD 6/19/00 30310.7 26340.0 19568FJF0,G5-3.2MKD-GD0 0004044 061900AD 6/19/00 24160.4 20995.4 19569F.F0.G5-3.2MKD-GD0 0004045 061900AD 6/19/00 27052.1 23508.3 19570F,F0,G5-3.2MKD-GD0 0004046 061900AD 6/19/00 19671.6 17094.6 19571F,F0,G5-3.2MKD-GD0 0004047 061900AD 6/19/00 29872.2 25958.9 19572FJ0.G5-3.2MKD-GD0 0004048 061900AD 6/19/00 25395.0 22068.3 19573F,F0,G5-3.2MKD-GD0 0004049 061900AD 6/19/00 23006.0 19992.2 19574F.F0.G5-3.2MKD-GD0 0004050 061900AD 6/19/00 29300.3 25462.0 19575F,F0,G5-3.2MKD-GD0 0004051 061900AD 6/19/00 23037.0 20019.2 19576F,F0,G5-3.2MKD-GD0 0004052 061900AD 6/19/00 36326.7 31567.9 19577F,F0,G5-3.2MKD-GD0 0004053 061900AD 6/19/00 34238.9 29753.6 19578F,F0,G5-3.2MKD-GD0 0004054 061900AD 6/19/00 34808.0 30248.2 19579F.F0.G5-3.2MKD-GD0 0004055 061900AD 6119/00 25356.7 22035.0 19580FJF0.G5-3.2MKD-GD0 0004056 061900AD 6/19/00 24851.5 21596.0 AVERAGE: 27524.1 23918.5 STANDARD DEVIATION: 4781 4155 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 26 of 103 Page 274 3M Medical Department Study: T-6295.12 ; Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Table VII. Summary of PFOS residues in Gl-Control on GD7 Sponsor ID 19501F,FO,G1-Control-GD7 19502F,F0,G1-Control-GD7 19504FJ0,G 1-Control-GD7 19505FJ'O.G1-Control-GD7 19506F,F0,Gl-Control-GD7 19507F,F0,G1-Control-GD7 19508F,F0,Gl-Control-GD7 19509F,F0,Gl-Control-GD7 1951OF,FO,G1-Control-GD7 1951IF,FO,G1-ontrol-GD7 19512F,FO,G1-Control-GD7 19513FJFO,G1-Control-GD7 19514FJFO,G1-Control-GD7 19515F,F0,G1-ControI-GD7 19516FJO.G1-Control-GD7 Centre ID Set Corrected Date Residue Found Residue Found Number Extracted (ne/e) (na/s0 0004057 062000A .6/20/00 ND ND 0004058 062000A 6/20/00 NQ NQ 0004059 062000A 6/20/00 NQ NQ 0004060 062000A 6/20/00 ND ND 0004061 . 062000A 6/20/00 17.8 15.5 0004062 062000A 6/20/00 ND ND 0004063 062000A 6/20/00 0.3 0.3 0004064 062000A 6/20/00 ND ND 0004065 062000A 6/20/00 ND ND 0004Q66 062000AR 6/20/00 ND ND 0004067 062000AR 6/20/00 ND ND 0004068 062000A 6/20/00 ND ND 0004069 062000A 6/20/00 ND ND 0004070 062000AR 6/20/00 ND ND 0004071 062000AR 6/20/00 NO NO JAVERAGE: 1.2 1.0 STANDARD DEVIATION: 5 4 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 27 of 103 Page 275 3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Table VIII. Summary of PFOS residues in G2-0.1MKD on GD7 Sponsor Centre ID Set Corrected Date Residue Found Residue Found ID Number Extracted (ne/g) (ng/f0 19517F,F0,G2-0.1MKD-GD7 0004072 062100A 6/21/00 409.4 355.8 19518F,F0,G2-0.1MKD-GD7 0004073 062100AD 6/21/00 561.4 487.9 19519FJ0.G2-0.1MKD-GD7 0004074 062100A 6/21/00 344.6 299.5 19521FF0.G2-0.1MKD-GD7 0004075 062100A 6/21/00 464.6 403.7 19522F,F0,G2-0.1MKD-GD7 0004076 062100AD 6/21/00 778.0 676.1 19523FJ0.G2-0.1MKD-GD7 0004077 062100AD 6/21/00 624.6 542.8 19524F,F0,G2-0.1MKD-GD7 0004078 062100AD 6/21/00 575.5 500.1 19525F,F0,G2-0.1MKD-GD7 0004079 062100AD 6/21/00 594.6 516.7 19526F.F0.G2-0.1MKD-GD7 0004080 062100A 6/21/00 469.2 407.7 19527F.F0.G2-0.1MKD-GD7 0004081 062100AD 6/21/00 741.9 644.7 19528F.F0.G2-0.1MKD-GD7 0004082 062100AD 6/21/00 531.8 462.1 19529F,F0,G2-0.1MKD-GD7 0004083 062100AD 6/2 1/0 0 744.4 646.9 19530F,F0,G2-0.1MKD-GD7 0004084 062100A m um 424.3 368.7 19531F.F0.G2-0.1MKD-GD7 0004085 062100AD 6/21/00 623.9 542.2 19532F.F0.G2-0.1MKD-GD7 0004086 062100AD 6/21/00 573.9 498.7 AVERAGE: 564.1 490.2 STANDARD DEVIATION: 128 111 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 28 of 103 Page 276 3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Table IX. Summary of PFOS residues in G3-0.4MKD on GD7 Sponsor Centre ID Set Corrected Date Residue Found Residue Found ID Number Extracted (na/s) (ng/g) 19533F.F0.G3-0.4MKD-GD7 0004087 062200AD 6/22/00 2548.5 2214.6 19534FJF0.G3-0.4MKD-GD7 0004088 062200AD 6/22/00 2693.6 2340.7 19535F.F0.G3-O.4MKD-GD7 0004089 062200AD 6/22/00 1824.8 1585.8 19536F,F0,G3-0.4MKD-GD7 0004090 062200AD 6/22/00 3261.1 2833.9 19537F,F0,G3-0.4MKD-GD7 0004091 062200AD 6/22/00. 2611.1 2269.0 19538F,F0,G3-O.4MKD-GD7 0004092 062200AD 6/22/00 2934.3 2549.9 19539F.F0.G3-0.4MKD-GD7 0004093 062200AD 6/22/00 2093.4 1819.2 19540F,F0,G3-0.4MKD-GD7 0004094 062200AD 6/22/00 2267.8 1970.7 19542F.F0.G3-0.4MKD-GD7 0004095 062200AD 6/22/00 3003.0 2609.6 19543FJ0.G3-0.4MKD-GD7 0004096. 062200AD 6/22/00 2797.7 '2431.2 19544FJFO,G3-0.4MKD-GD7 0004097 062200AD 6/22/00 2284.0 1984.8 19546FJF0.G3-0.4MKD-GD7 0004098 062200AD 6/22/00 1523.1 1323.6 19547FJ0.G3-0.4MKD-GD7 0004099 062200AD 6/22/00 2055.6 1786.3 19548F,F0,G3-0.4MKD-GD7 0004100 062200AD 6/22/00 2847.6 2474.6 AVERAGE: 2481.8 2156.7 STANDARD DEVIATION: 492 428 Centre Analytical Laboratories, Inc. 3IVI Environmental Laboratory Page 29 of 103 Page 277 3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Table X. Su m m a ry of PFOS residues in G4-1.6MKD on GD7 Sponsor Centre ID Set Corrected Date Residue Found Residue Found ID Number Extracted (na/s) (ng/g) 19550FJF0.G4-1.6MKD-GD7 0004101 062200BD 6/22/00 14977.5 13015.4 19553FJ0,G4-1.6MKD-GD7 0004102 062200BD 6/22/00 12262.8 10656.4 19554F,F0,G4-1.6MKD-GD7 0004103 062200BD 6/22/00 11777.1 10234.3 19555FJF0.G4-1.6MKD-GD7 0004104 062200BD 6/22/00 9607.3 8348.7 19556F,F0,G4-1.6MKD-GD7 0004105 062200BD 6/22/00 10222.3 8883.2 19558F,F0,G4-1.6MKD-GD7 0004106 062200BD 6/22/00 5886.5 5115.4 19559F,F0,G4-1.6MKD-GD7 0004107 062200BD 6/22/00 6894.8 5991.6 19560F,F0,G4-1.6MKD-GD7 0004108 062200BD 6/22/00 7024.2 6104.0 19561FJF0.G4-1.6MKD-GD7 0004109 062200BD 6/22/00 8877.7 7714.7 19562F,F0,G4-1.6MKD-GD7 0004110 062200BD 6/22/00 10845.1 9424.4 19563FJ0.G4-1.6MKD-GD7 0004111 062200BD 6/22/00 14910.3 12957.1 19564F.F0.G4-1.6MKD-GD7 0004112 062200BD 6/22/00 13656.0 11867.1 AVERAGE: 10578.5 9192.7 STANDARD DEVIATION: 3076 2673 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 30 of 103 Page 278 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Table XI. Summary of PFOS residues in G5-3.2MKD on GD7 Sponsor Centre ID Set Corrected Date Residue Found Residue Found ID Number Extracted (ng/j?) (ne/g) 19565F,F0,G5-3.2MKD-GD7 0004113 062300AC1 6/23/00 51657.5 44890.4 19566FJF0.G5-3.2MKD-GD7 :0004114 062300AD 6/23/00 26453.8 22988.4 19567FJF0.G5-3.2MKD-GD7 0004115 062300AD 6/23/00 35913.2 31208.6 19568F,F0,G5-3.2MKD-GD7 0004116 062300AD 6/23/00 23007.4 . 19993.4 19569FJF0,G5-3.2MKD-GD7 0004117 062300AD 6/23/00 42878.2 37261.2 . 19570F,F0,G5-3.2MKD-GD7 0004118 062300AD 6/23/00 37368.9 32473.6 19571FJ0,G5-3.2MKD-GD7 0004119 062300AD 6/23/00 42995.9 37363.4 19572F.F0.G5-3.2MKD-GD7 0004120 062300AD 6/23/00 38826.6 33740.3 19573FJF0.G5-3.2MKD-GD7 0004121 062300AD 6/23/00 26685.9 23190.0 19574FJ0.G5-3.2MKD-GD7 0004122 062300AD 6/23/00 36333.3 31573.6 19575FJ0,G5-3.2MKD-GD7 0004123 062300AD 6/23/00 35455.7 30811.0 19577FJ0.G5-3.2MKD-GD7 0004124 062300AD 6/23/00 68339.2 59386.8 19579FJF0.G5-3.2MKD-GD7 0004125 062300AD 6/23/00 35856.5 31159.3 19580F,F0,G5-3.2MKD-GD7 0004126 062300AD 6/23/00 30065.5 26126.9 AVERAGE: 379S8.4 33011.9 STANDARD DEVIATION: 11516 10008 Centre Analytical Laboratories, Inc. 3IVI Environmental Laboratory Page 31 o f 103 Page 279 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Table XII. Summary of PFOS residues in Gl-Control on GD15 Sponsor Centre ID Set Corrected Date Residue Found Residue Found ID Number Extracted (ne/g) (na/e) 19501F,F0,Gl-Control-GD15 0004127 062300B 6/23/00 ND 0.0 19502F,F0,G1-Control-GD15 0004128 062300B 6/23/00 ND 0.0 19504FJF0,Gl-Control-GD15 0004129 062300B 6/23/00 ND 0,0 19505F,FO,G1-Control-GD15 0004130 062300B 6/23/00 ND 0.0 19506FJF0,Gl-Control-GD15 0004131 062300B 6/23/00 44.5 38.7 19507F,F0,G1-Control-GD 15 0004132 0623OOB 6/23/00 ND 0.0 19508FJ0,Gl-Control-GD15 0004133 062300B 6/23/00 ND 0.0 19509FJF0,Gl-Control-GD15 0004134 062300B 6/23/00 24.2 21.0 195lOFJO.Gl-Control-GD 15 0004135 062300B 6/23/00 7.9 6.9 19511F,FO,G1-Control-GD15 0004136 062300B 6/23/00 ND 0.0 19512F,F0,Gl-Control-GD 15 0004137 062300B 6/23/00 57.9 50.3 19513F,F0,Gl-Control-GD15 0004138 062300B 6/23/00 ND 0.0 19514FJFO,G1-Control-GD15 0004139 062300B 6/23/00 ND 0.0 19515FJF0,Gl-Control-GD15 0004140 062300B 6/23/00 ND 0.0 19516FJF0.Gl-Control-GD15 0004141 062300B 6/23/00 ' AVERAGE: 21.6 10.4 18.6 9.0 STANDARD DEVIATION: 19 16 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 32 o f 103 Page 280 3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Table XIII. Summary of PFOS residues in G2-0.1MKD on GD15 Sponsor Centre ID Set Corrected Date Residue Found Residue Found ID Number Extracted (ne/g) (ng/g) 19517F,F0,G2-0.1MKD-GD15 0004142 062600AD 6/26/00 758.1 658.8 19518F,F0,G2-0.1MKD-GD15 0004143 062600AD 6/26/00 671.8 583.8 I9519FJF0,G2-0.1MKD-GD15 0004144 062600AD 6/26/00 782.2 679.7 19521FJ0,G2-0.1MKD-GD15 0004145 062600AD 6/26/00 632.3 549.5 19522FJF0.G2-0.1MKD-GD15 0004146 062600AD 6/26/00 817.4 710.3 19523F,F0,G2-0.1MKD-GD15 0004147 062600AD 6/26/00 641.7 557.6 19524F.F0.G2-0.1MKD-GD15 0004148 062600AD 6/26/00 834.2 724.9 19525FJF0,G2-O.lMKD-GD15 0004149 062600AD 6/26/00 1080.8 939.2 19526FJF0.G2-0.1MKD-GD15 0004150 062600AD 6/26/00 777.9 676.0 19527FJF0.G2-0.1MKD-GD15 0004151 062600AD m em 798.5 693.9 19528F,F0,G2-0.1MKD-GD15 0004152 062600AD m em 844.8 734.1 19529F,F0,G2-0.1MKD-GD15 0004153 062600AD m em 723.6 628.8 19530F,F0,G2-0.1MKD-GD15 0004154 062600AD1 6/26/00 724.4 629.5 19531F.F0.G2-0.1MKD-GD15 0004155 062600AD 6/26/00 777.3 675.5 19532F.F0.G2-0.1MKD-GD15 0004J56 062600AD 6/26/00 568.2 493.8 AVERAGE: 762.2 662.4 STANDARD DEVIATION: 119 104 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 33 o f 103 Page 281 3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Table XIV. Summary of PFOS residues in G3-0.4MKD on GD15 Sponsor Centre ID Set Corrected Date Residue Found Residue Found ID Number Extracted (na/g) (ng/g) 19533F,F0,G3-0.4MKD-GD15 0004157 062600BD 6/26/00 2759.7 2398.2 19534F.F0.G3-0.4MKD-GD15 0004158 062600BD 6/26/00 4817.3 : 4186.2 19535F.F0.G3-0.4MKD-GD15 0004159 062600BD 6/26/00 3627.9 3152.6 19536FJ0.G3-0.4MKD-GD 15 0004160 062600BD 6/26/00 3186.8 2769.3 19537F,F0,G3-0.4MKD-GD15 0004161 062600BD 6/26/00 3397.0 2952.0 19538F,F0,G3-0.4MKD-GD15. 0004162 062600BD 6/26/00 3143.5 2731.7 19539FJ0.G3-0.4MKD-GD 15 0004163 062600BD 6/26/00 3094.1 2688.8 19540F,F0,G3-0.4MKD-GD15 0004164 062600BD 6/26/00 4151.1 3607.3 19542F.F0.G3-0.4MKD-GD15 0004165 062600BD 6/26/00 1881.6 1635.1 19543F,F0,G3-0.4MKD-GD15 0004166 062600BD 6/26/00 3420.1 2972.1 19544F,F0,G3-0.4MKD-GD15 0004167 062600BD 6/26/00 3372.6 2930.8 19546F,F0,G3-0.4MKD-GD15 0004168 062600BD 6/26/00 2625.1 2281.2 19547FJO.G3-0.4MKD-GD15 0004169 062600BD 6/26/00 3863.8 ' 3357.6 19548FJ0.G3-0.4MKD-GD 15 0004170 062600BD 6/26/00 3910.6 3398.3 - AVERAGE: 3375.1 2932.9 STANDARD DEVIATION: 713 620 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 34 of 103 Page 282 3IVI Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 . Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Table XV. Summary of PFOS residues in G4-1.6MKD on GD15 Sponsor Centre ID Set Corrected Date Residue Found Residue Found ID Number Extracted 19550FJF0,G4-1.6MKD-GD15 0004171 062700AD 6/27/00 (nBfa) 13868.2 (ne/g) 12051.5 19553FJ0.G4-1.6MKD-GD15 0004172 062700AI) 6/27/00 14838.7 12894.8 19554F,F0,G4-1.6MKD-GD15 0004173 062700AD 6/27/00 14172.4 12315.8 19555F.F0.G4-1.6MKD-GD15 0004174 062700AD 6/27/00 16258.1 14128.3 19556FJF0.G4-1.6MKD-GD15 0004175 062700AD 6/27/00 9066.1 7878.4 19558FJ0.G4-1.6MKD-GD15 0004176 062700AD 6/27/00 7174.9 6235.0 19559FJF0.G4-1.6MKD-GD15 0004177 062700AD 6/27/00 7477.4 6497.9 19560F.F0.G4-1.6MKD-GD15 0004178 062700AEt 6/27/00 14880.9 12931.5 19561F.F0.G4-1.6MKD-GD15 0004179 062700AD 6/27/00 17442.7 15157.7 19562FJ0.G4-1.6MKD-GD15 0004180 062700AD 6/27/00 11417.2 9921.5 19563F,F0,G4-1.6MKD-GD15 0004181 062700AD 6/27/00 8949.0 7776.7 19564F,F0,G4-1.6MKD-GD15 0004182 062700AE1 6/27/00 17342.4 15070.5 AVERAGE: 12740.7 11071.6 STANDARD DEVIATION: 3769 3275 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 35 of 103 Page 283 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Table XVI. Summary of PFOS residues iin G5-3.2MKD on GDIS Sponsor Centre ID Set Corrected Date Residue Found Residue Found ID Number Extracted (ng/g) (ng/g) 19565F,F0,G5-3.2MKD-GD15 0004183 062800AD 6/28/00 35175.8 30567.8 19566FJF0.G5-3.2MKD-GD15 0004184 062800AD 6/28/00 15192.6 13202.4 19567FJ?0,G5-3.2MKD-GD15 0004185 062800AD 6/28/00 20391.7 17720.4 19568F,F0,G5-3.2MKD-GD15 0004186 062800AD 6/28/00 46897.0 40753.5 19569F,F0,G5-3.2MKD-GD15 0004187 062800AD 6/28/00 36307.7 31551.4 19570F,F0,G5-3.2MKD-GD15 0004188 062800AD 6/28/00 31052.5 26984.6 19571F,F0,G5-3.2MKD-GD15 0004189 062800AD 6/28/00 28440.6 24714.9 19572FJ0.G5-3.2MKD-GD15 0004190 062800AD 6/28/00 20549.6 17857.6 19573FJ0.G5-3.2MKD-GD15 0004191 062800AD 6/28/00 39763.7 34554.7 19574FJ:0,G5-3.2MKD-GD15 0004192 062800AD 6/28/00 47267.6 41075.5 19575F.F0.G5-3.2MKD-GD15 0004193 062800AD 6/28/00 40708.5 35375.7 19577F,F0,G5-3.2MKD-GD15 0004194 062800AD 6/28/00 47288.1 41093.4 19579F,F0,G5-3.2MKD-GD15 0004195 062800AD 6/28/00 33436.6 29056.4 19580F.F0.G5-3.2MKD-GD15 0004196 062800AD 6/28/00 32709.0 28424.1 AVERAGE: 33941,5 29495.2 STANDARD DEVIATION: 10248 8905 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 36 of 103 Page 284 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 , Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Table XVII. Summary of PFOS residues in Gl-Control on GD21 Sponsor Centre ED Set Corrected Date Residue Found Residue Found ID Number Extracted (ng/g) (ng/g) 19504F,F0,Gl-Control-GD21 0004197 062900A 6/29/00 ND ND 19505FJF0.G1-Control-GD21 0004198 062900A 6/29/00 ND ND 19506FrF0,Gl-Control-GD21 0004199 062900A 6/29/00 12.3 10.7 19507FJF0.G1-ControI-GD21 0004200 062900A 6/29/00 ND ND 19509F,F0,G1-Control-GD21 0004201 062900A 6/29/00 84.3 73.3 19512FJ0,G 1-Control-GD21 0004202 062900A 6/29/00 21.5 18.7 19516F.F0.G1-Control-GD21 0004203 062900A 6/29/00 2.0 1.7 AVERAGE: 17.2 14.9 STANDARD DEVIATION: 31 27 Table XVUI. Summary of PFOS residues in G2-0.1MKD on GD21 Sponsor Centre ID Set Corrected Date Residue Found Residue Found ID Number Extracted (ng/g) . (ng/g) 19518FJF0.G2-0.1MKD-GD21 0004204 0629-3000AD 6/29/00 735.4 639.1 19519FE0.G2-0.1MKD-GD21 0004205 062900A 6/29/00 473.0 411.0 1952IFE0,G2-0.1MKD-GD21 0004206 062900A m a m 403.4 350.6 19523FJF0.G2-O.1MKD-GD21 0004207 062900A 6/29/00 402.1 349.4 19524F.F0.G2-0.1MKD-GD21 0004208 062900A 6/29/00 493.9 429.2 19528F.F0.G2-0.1MKD-GD21 0004209 062900A 6/29/00 456.1 396.4 19529F.F0.G2-0.1MKD-GD21 0004210 062900A 6/29/00 391.4 340.1 AVERAGE: 479-3 416.5 STANDARD DEVIATION: 120 104 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 37 of 103 Page 285 3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 , Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Table XLX. Summary of PFOS residues in G3-0.4MKD on GD21 Sponsor Centre ID Set Corrected Date Residue Found Residue Found ID Number Extracted (ne/g) (ne/g) 19533F,F0,G3-O.4MKD-GD21 0004211 0629-3000AD 6/30/00 1684.4 1463.7 19534FVF0,G3-0.4MKD-GD21 0004212 0629-3000i\D 6/30/00 3102.9 2696.4 19536FF0.G3-O.4MKD-GD21 0004213 0629-3000AD 6/30/00 1848.1 1606.0 19543F.F0.G3-O.4MKD-GD21 0004214 0629-3000/VD 6/30/00 2233.1 1940.6 19546F,F0,G3-O.4MKD-GD21 0004215 0629-3000AD 6/30/00 2229.2 1937.2 19548F,F0,G3-0.4MKD-GD21 0004216 0629-3000D 6/30/00 5417.3 4707.6 AVERAGE: 2752.5 2391.9 STANDARD DEVIATION: 1395 1212 Table XX. Summary o f PFOS residues in G4-1.6MKD on GD21 Sponsor Centre ID Set Corrected Date Residue Found Residue Found ID Number Extracted (ng/g) (ng/g)....... 19556F.F0,G4-1.6MKD-GD21 0004217 0629-3000AD 6/30/00 18272.7 . 15879.0 19559FJF0.G4-1.6MKD-GD21 0004218 0629-3000AD 6/30/00 10574.8 9189.5 19560F,F0,G4-1.6MKD-GD21 0004219 0629-3000AD 6/30/00 11190.1 9724.2 19562F,F0,G4-1.6MKD-GD21 0004220 0629-3000AD 6/30/00 5681.8 4937.5 AVERAGE: 114293 9932.5 STANDARD DEVIATION; 5185 4506 Table XXI. Summary of PFOS residues in G5-3.2MKD on GD21 Sponsor Centre ID Set Corrected Date Residue Found Residue Found ID Number Extracted . (ng/g) (ng/g) 19568F.F0.G5-3.2MKD-GD21 0004221 0629-3000AD 6/30/00 25084.0 21798.0 19569F,F0,G5-3.2MKD-GD21 0004222 0629-3000AD 6/30/00 21667.4 18829.0 19571FJ0.G5-3.2MKD-GD21 0004223 0629-3000AD 6/30/00 21465.5 18653.5 19572F,F0,G5-3.2MKD-GD21 0004224 0629-3000AD 6/30/00 15006.4 13040.6 19577F,F0,G5-3.2MKD-GD21 004225 0629-3000AD 6/30/00 27066.6 23520.9 19579F,F0,G5-3.2MKD-GD21 0004226 0629-3000AD 6/30/00 28290.3 24584.3 AVERAGE:: 23096.7 20071.0 STANDARD DEVIATION: 4834 4201 Centre Analytical Laboratories, Inc. Page 38 of 103 3M Environmental Laboratory Page 286 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Table XXII. Summary of PFOS recoveries in Fortified Samples Sponsor ID 50 IFJO,Gl-Control-GDO 504F,F0,G1-Control-GDO ^05FJO,Gl-Control-GDO 07F,F0,Gl-Control-GD0 11F,F0,Gl-Control-GDO 2F.F0,Gl-Control-GDO 3FJO,Gl-Control-GDO 5FJ0.G 1-Control-GDO IFJO,Gl-Control-GDO 1 IFJ0,G1-Control-GDO 1 >FJ0,Gl-Control-GDO 1! FJO,Gl-Control-GDO IS FJO,G1-Control-GDO 19 FJO,Gl-Control-GDO 19. 7J0,G 1-Control-GD7 19i '.F0,Gl-Control-GD7 19507x. T,Gl-Control-GD7 19509FJ0,Gl-Control-GD7 19510FJ0,Gl-Control-GD7 1951IFJ0.G1 -Control-GD7 19512FJO,G1-Control-GD7 19513FJ0.G1 -Control-GD7 19514FJO,G1-Control-GD7 19516FJO,G1-Control-GD7 1950IFJO,G1-Contiol-GD7 19505FJ0,Gl-Control-GD7 19501FJ0,Gl-Control-GD15 19502FJ0,Gl-Control-GD15 19504FJ0,Gl-Control-GD15 19505FJ0,Gl-Control-GD15 19507FJ0,Gl-Control-GD15 19508F,F0,G1-Control-GD15 19513FJ0,Gl-Control-GD 15 19514FJO.Gl-Control-GD 15 Centre Set Date Ana. % ID Number Extracted Added (ng) Recovery 0003977 Spk A 061200AR 6/12/00 50 126 0003980 SpkB 61200AR 6/12/00 250 116 0003981 Spk A 061400A 6/14/00 50 98 0003983 SpkB 061400A 6/14/00 250 97 0003987 Spk A 061500AD 6/15/00 50 128 0003988 SpkB 061500AD 6/15/00 250 110 0003989 Spk A 061600A 6/16/00 50 80 0003991 SpkB 061600A 6/16/00 1000 89 0003986 Spk A 061900AD 6/19/00 50 107 0003987 SpkB 061900AD 6/19/00 5000 78 0003986 Spk C 062000A 6/20/00 50 128 0003987 Spk D 062000A 6/20/00 20000 96 0003989 Spk A 062100AD 6/21/00 1000 104 0003991 SpkB 062100A 6/21/00 20000 80 0004057 Spk A 062200A 6/22/00 1000 97 0004060 SpkB 062200A 6/22/00 20000 79 0004062 Spk A 062200B 6/22/00 1000 99 0004064 SpkB 062200B 6/22/00 20000 86 0004065 Spk A 062:300A 6/23/00 1000 93 0004066 Spk B 062:3OOA 6/23/00 20000 71 0004067 Spk A 062300B 6/23/00 1000 85 0004068 SpkB 062300B 6 /23/0 0 20000 81 0004069 Spk A 062()00A 6/26/00 50 111 0004071 SpkB 062600A 6/26/00 1000 94 0004057 Spk C 062600B 6/26/00 50 82 0004060 Spk D 062600B 6/26/00 2000 100 0004127 Spk A 062700A 6/27/00 100 100 0004128 SpkB 062700A 6/27/00 20000 100 0004129 Spk A 062800A 6/28/00 500 99 0004130 SpkB 0628OOA 6/28/00 100000 89 0004132 Spk A 062900A 6/29/00 50 119 0004133 SpkB 0629OOA 6/29/00 2000 95 0004138 Spk A 063C00A 6/30/00 500 113 0004139 SpkB 063000A 6/30/00 20000 85 AVERAGE: 98 STANDARD DEVIATION: 15 RELATIVE STANDARD DEVIATION: 15 Centre Analytical Laboratories, Inc. Page 39 of 103 3M Environmental Laboratory Page 287 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 . Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 12.0 FIGURES Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 40 of 103 Page 288 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Figure 1. . . ' Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Typical Calibration Curve for PFOS Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 41 of 103 Page 289 3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 Figure 2. Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Typical Mean Response Factor for THPFOS . Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 42 of 103 Page 290 3M Medical Department Study. T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 Figure 3. Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Chromatogram Representing a 5 ng/mL extracted standard for PFOS and 250 ng/mL fortification of THPFOS 1: PFOS XC06120-1, 5 ng/mL Standard" 061200AR-2002 Sm (SG, 2x2) 10 0 -, . . 52 518:n 13-Jun-2QQ0 09:00:24 LC/MS/MS #6 MRMof2 Channels ES499 >99 6.24e4 Area 1.0 2.0 ' 3330 2: THPFOS___________________ XC061200-1, 5 ng/mL Standard 061200AR-2002 Sm (SG, 2x2) 100-, %-l J? 4.00 5.00 V .. 6.00 5.11 923071 7.00 8.00 Time 13-Jun-2Q00 09:00:24 LC/MS/MS #6 MRM of 2 Channels ES- 427 >80 8.67e5 Area 1.00 2.00 3.00 4.0 5. ' ' 6.0 7.00 8.00 Time Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 43 of 103 Page 291 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Figure 4. Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Chromatogram Representing a 125 ng/mL extracted standard for PFOS and 250 ng/mL fortification of THPFOS '-S C 2U-5,125 ng/mL Standard '1 'AR-2006 Sm (SG. 2x2) 10- %- 5,26 101.957 13-Jun-2Q 00 10:07:191 -w,, LC/MS/MS MRM of 2 Channels ES- 499 > 99 8.76e5 Area 1.G0 2.00 3.00 2: THPFOS______ Xuo61200-5,125 ng/mL Standard 061200AR-2006 Sm (SG. 2x2) : 10 0 -, 4.0 5.00 5.10 1007191 6.00 % l 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 Time iJ-Jun-2000 10:07:19 . LC/MS/MS #6 MRM of 2 Channels ES- 427 > 80l 8.93e^ Areai 7.00 8.0 Time Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 44 of 103 Page 292 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Figure 5. Centre Study No.: 023-016 Sponsor Protocol No: F A C T -T O X -1 10 Chromatogram Representing Control Rat Feces for PFOS and THPFOS (Centre ED: 0004277 Blank B, Set: 062300A) 1 : PFOS Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 45 of 103 Page 293 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Figure 6. Chromatogram Representing Control Rat Feces Fortified with 50 ng of PFOS and 500 ng of THPFOS (Centre ID: 0003981 Spk A, Set: 061400A) THPFOS 1003981 SpkA '6140QA-1013 Sm (SG, 2x2) 100-1 5.10 91634^ 14-Jun-2000 22:08:45 LC/MS/MS #6 MRM of 2 Channls ES427 > 80 7.84e5 Area 1.00 ' 2.00 ' 3.00 4.00 5.00 6.00 7.00 Time Centre Analytical Laboratories, Inc. 3IVI Environmental Laboratory Pag 46 of 103 Page 294 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Figure 7. Centre Study No.: 023-016 Sponsor Protocol No: F A C T -T O X -110 Chromatogram of Rat Feces Srnnple from G2 on GDO (Centre ID: 0003994,Set: 061400A) 2 : TH9FOS 0003994 061400A-1018 Sm (SG, 2x2) 10On 5.07 748991 14-Jun-2Q00 23:32:39 LC/MS/MS #6 MRM of 2 Channels ES427 > 60 6.42e= Area 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 Time Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 47 of 103 Page 295 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FA C T -T O X -110 13.0 APPENDICES Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 48 of 103 Page 296 3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 Centre Study No.: 023-016 S ponsor Protocol No: F A C T -T O X -l 10 APPENDIX A Study Protocol FACT-TOX-110 (Centre Study No. 023-016) and Amendments and Deviations Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 49 of 103 Page 297 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 . ^ enVt'^0nm<nla*Technology and Services Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 PO Sox 33331 Si.Pjol.M NiJI3 . 3.3 3 3 1 612 773 5442 P n tc c z l ZFACT-7O X-110 3M S tu dy Title . Oral (Gavage) Pharmacokinetic Study of PFQS in Rats PROTOCOL Author Lisa Clemea Date: JuneS, 1999 Performing Laboratory 3M Environmental Technology & Safety Services . 3M Environmental Laboratory . 935 Bush Avenue . St. Paul, MN 55106 LaboratoryProject Identification FACT-TOX-110 U2S49 Copyoforigin 3WE nvironm ental Laboratory Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 1 o f to / Page 50 of 103 Page 298 3M Medical Department Study: T-6295.12 i; Analytical Report: FACT TOX-110 LRN-U2849 ' Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Protocol HFAGT-TOX-1 10 Sttidy Identification Oral (Gavage) Pharmacokinetic Study o f PFOS in Rats . T est Material Perfluoroodane sulfonic acid potassium salt (T-6295) Sponsor . Sponsor Representative S tu dy Director 3M Toxicology Services - Medical Department 3M Center, Building 220-2E-02 . S t Paul, MN 55144-1000 '_' Marvin T. Case, D.V.M., Ph T) 3M Toxicology Services Telephone: 651-733-5180 Facsimile: 651-733-1773 ' ' Kristen I Hansen, PhD. 3M Environmental Technology and Safety Services Building 2-3E-09. 651-778-6018 .. . . S tu d y Location(s) . In vivo Testing Fadfity Analytical Testing Laboratory Argus Research Laboratories, Inc. 905 Sheehy Drive, Building A Horsham, PA 19044 3M Environmental Laboratory Building 2-3E-C9 935 Bush Avenue S t Paul, MN 55106 ' 3M Environm ental Laboratory ' Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 2 o f 10 . Page'51 of 103 Page 299 3M Medical Department Study: T-6295.12 Sub-Contract Laboratory P ro p o sed Stu dy Timetable Study Initiation Date Study Completion Date Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Protocol ttrACT-TOX-1 10 Advanced Bioanalyrical Services, Jsc. 15 Catbenvcod Road Ithaca, NY 14850 1 Battelle Memorial Institute 505 King Avenue Columbus, Ohio 43201-2693 June 8,1999 August 8,2000 1. S tudy ' ' .' , Oral (gavage) pharmacokinetic study ofpotassium perfluorocctsne sulfonic acid (PFOS) in rats. 2. P urpose This analytical study is designed to determine levels ofpotassiumperilnorooctanesulfcnate (PFOS) in specimens ofliver and serum of rats. The in-life portion of this study was conducted at Argus Researehiaboratories, study 3418-013. All serum samples will be extracted and analyzed at Advanced Bioanalyrical Services, Inc. and all liver simples extracted and analyzed at Battelle Memorial Institute. Additional analyses may be performed at the 3M Environmental Laboratory as methods are developed and validated. If additional analyses are performed an amendment to this protocol will be written. " ' 3 . R e g u l a t o r y Co m plian ce . This study will be conducted in accordance with the United States: Food and Drug Administration, Good Laboratory Practices Standards, .Final Rule 21 CFR 58, with the exception that analysis of the test material mixture for concentration, solubility, homogeneity, and stability will not he conducted,and is die responsibility o f the Sponsor. ' 4 . Q u a lit y A ssu r a n c e The 3M Environmental Laboratory Quality Assurance Unit will view the protocol and audit study conduct, data, and final report to determine compliance with Good Laboratory Practice Standards and with 3M Environmental Laboratory Standard Opearing Procedures. The QA Unit at the sub-contract laboratory will audit their studyconduct, data, and results report prior to submitting to the 3M Environmental Laboratory. 3M E nvironm ental Laboratory Centre Analytical Laboratories, Inc. 3IVI Environmental Laboratory . P a g e 3 o f 10 . Page'52 of 103 Page 300 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 . LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-LIO Protocol SFAC T-TO X-110 5. TES1TMaTERIAL 6.1 Refer toArgus Research Laboratory protocol for study #418-013. 6. C o n tr o l Ma tr ic e s 6.1 Identification Rat liver and serum and/or rabbit liver and serum, traceability numbers will be recorded in the raw data and included in the final report 6.2 Source Argus Research and/or Sigma Chemical 6.3 P h ysical Description Rat liver and serum and/orrabbit liver and serum 6.4' Purity and Stability Not applicable . . 6.5 S torage Conditions Frozen at -20 C 10 C or-50C 10 "C 6.6 R eserve Matrix A portion of the control matrix will be retained in' the 3M archives. for as long as the quality o f the preparation affords evaluation, but not longer than ten . years following the effective date o f the final test rule (ifapplicable). 6.7 D isposition Matrices will be retained at the 3M Environmental Laboratory per GLP regulation. Certain matrices (feces, urines and blood) may be disposed after QAU verification. ' 6.8 S a fety Precautions Refer to MSDS for chemicals used. Wear appropriate ' laboratory attire, and follow adequate precautions for handling biological materials and preparing samples for analysis. : ` 7. Reference Material 7.1 identification Potassium perfluorooctanesulfonate (PROS), lot #s 171,215, or 217 (equivalent lots) ' 7.2 S ource 3MJSpesialty Chemicals 7.3 P h ysical Description White powder 7.4 P urity and Stability Purity ofPFOS is 99% or greater. Stability has not been determined. ' . 7.5 S torage Conditions Room temperature 7.6 R ese rv e Materia! A reserve sample from each batch ofPFOS used in this study will be retained as long as the quality of the preparation affords evaluation, but not longer than ten years following the effective date of the final test rule (it applicable). 7.7 D isposition Unused reference material will be retained foruse by the 3M Environmental Laboratory and will be discarded when tire quality ofpreparation no longer affords evaluation. ' 3M Bivironmentat Laboratory Page 4 f10 Centre Analytical Laboratories, Inc, 3M Environmental Laboratory Page 53 of 103 Page 301 3M Medical Department Study. T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 . Sponsor Protocol No: FACT-TOX-110 Prtcal&ACT-TQX-no 7.8 S a fe ty P recautions Refer to MSDS for chemicals used. Wear approonate laboratory attire,'aad follow adequate precautions for handling biological materials and preparing samples for analysis. 8. Te s t S ystem Rats were used as the test system and were maintained and dosed as described in the Argus Research protocol #418-013. The female rats will be given the test material or control once daily beginning 42 days prior to cohabitation and continue through day 14 or 20 ofpresumed . " gestation. See table 1 for more dosage information. . . Table 1 _ -------------------- Dosage Levels, Concentration, and Volmes uosage Group t i 4 5 Number of female rats Id 16 - 16 16 16 . Dosage mg/kg/day 0 0.1 0.4 1 1.6 3.2 Concentration mgflegttay 0 0.02 0.08 022 .64 . uosage volume mlAg 51 " ------------"1 - 5 ---------1 9. S p e c i m e n m b S a m p l e R e c e i p t . The 3M Environmental Laboratory will restive homogeneity samples for dose analysis and specimens o f the following body tissues and fluids from the indicated points in the study. See table 2 for specimen information. All specimens will be packed on dry ice for shipping. . Table 2 . . Specimen Information Body tbsnc/Bnid Collected ' Expected# of specimens serum - Dam and Fetus animals Unne and feces--Dam anjmaig Liver --Darn and Fetus animals Milk Secreting Glands - Dam animals Anmtotic Fluid-Dam ^"uais . Dam-Predose, Days 7,15, and 21 Fetus-At lamination o f the study . Predose, Days 7,15, and 21 Dam and Fetus-At termination of the study At the termination of the study Day 15 and Day 21 320 Dam and 320 Fetus fnooledt 320 urine ami 320 feces #0 Dam and 80 Fetus (pooled 1 5 ---------- 1T --------- 3 M E nvironm ental Laboratory Pagesof to Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 54 of 103 Page 3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 ' LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Prcloeal1tFAC T.TO X.U 0 . .' * Table 2 C oni . Specimen Information t Body tissue/Huid fcmbryo's with Placentae - Dam animals ' U ircass-Fetos animal Placentae--Fetus animal . Collected Day 15 and Day 21 R e s te d # of specimens To ------- At the termination of the sntdv ~ 160 Day 15 and Day 21 8 ' :-- " j 2000 Total number o f test animals: 64 .Total num'ber o fcontrol anipiratr 16 . Specimens seat to 3M Environmental Laboratories will be received and tracked accormn? to applicable Standard OperatingProcedures. 10. Preparato ry Methods 10.1 FACT-M-1.1, Extraction ofPotassium Perfluorooctanesulfonate or OtherAnionic Fhorochemical Surfactant from Liver for Analysis Using HPLC-Electrospray/Mass Spectrometry ' .* 10.2 ETS-M .l, Extraction ofPotassium Perfluorooctanesulfonate or Other * Fluorochemical Compounds from Serum or Other Fluid for Analysis Using EPLC- Hectrospray/Mass Spectrometry ' :: 10.3 If preparatory methods other than those listed above rreused, an amendment to this protocol will be written. Any deviations from these methods will be documented and included with the studydata. . 10.4 If analyses are sub-contracted to other laboratories, an amendment will be written to . include theirmethods and copies ofeach method will be attached to this protocol 11. A n a lytic a l M eth o d s . 11-1 FACT-M-2.1, Analysis ofFluorochemicals in Liver Extracts Using HPLCElectrospray/Mass Spectrometry' 11-2 ETS-8-5.1, Analysis ofPotassiumPerfluorooctanesulfbnate or Other Fluorochemicals in Serum or Other Fluid Extracts Using HPLC-Eectrospiay/Mass Spectrometry 11.3 If analytical methods other than those listed above are used, an amenrfm^i t0 jy protocol will be written. Any deviations from these methods will be documented and included with the study data. '- ' 11-4 If analyses are sub-contracted to other laboratories, an amendment will he written to include their methods and copies ofeach method wiil be attached to this protccoL 3M Environm ental Laboratory Page 6 o f10 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 55 of 103 Page 303 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 . Protocol 4PACT-TCX-110 I Z D a t a Q u a lity Ob je c t iv e s The aumber o f spikes/duplicates, use of surrogates, and information on other data aualitv indicators are included in the analytical methods. In addition, ihe following criteria willbe me1- 1Z 1 Linearity ri i 0.93 . 12.2 Limits o f detection / quantitation 1 Z Z 1 Method Detection Limit (MDL) for PFOS . a) Serna: 1.75 ppb . .b) Liver 15 ppb . . 1 Z Z 2 Limit ofQuantitation (LOQ) - Equal to the lowest acceptable standard in the calibration curve 1 Z 3 Duplicate acceptable precisio n < 30% for the method ' 1Z4 Spike acceptable recoveries 70%-130% 12.5 U se o f confirm atory m eth o d s Indeterminate samples will be re-analvzed nriTM ,, confirmatory method. If a confirmatory method is used, an amendment to this protocol will be written. 12.6 D emonstration o f sp ecificity Chromatographic retention time, mass special daughter ion characterization. *' 13. S u b -C o n t r a c t e d A n a l y s i s .. 13.1 All analyses as detaed in this protocol will be performed at 3M Environmental Laboratories, Building 2-3E-09,935 Bush Avenue, St Paul, MN 55106, at Advanced Bioanalytical Services, Etc, 15 Catherwood Road, Ithaca, NY 14850, or at Battelle Memorial Institute, 505 King Avenue, Columbus, Ohio 43201-2693. 13.2 An amendment to this protocol will be written if analyses are performed at . laboratories other than 3M Environmental Laboratories, AdvancedBioanalytical Services, Inc., orBattelleMemorial Institute. 14. STAT7ST1CALANALYSIS Averages and standard deviations will be calculated. The statistical methods that will be used art described below: ' 14.1 Data transformations a n d analysis Data will be reported as the concentration (weight/weight orweisht/vol) ofPFOS or metabolite per tissue or fluid. 3ME nvironm ental Laboratory Centre Analytical Laboratories, Inc. 3M Environmental Laboratory P a g e 7 o f 10 Page 56 of 103 Page 304 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 P r e ta x ! F A C T -7 O X-110 14.2 Statistical analysis Statistics used may include regression analysis of concentrations over time, and standard deviations calculated for the concentrations within each dose group. Ifnecessary, simple statistical tests, such as Student's t test, may be applied to evaluate statistical diuerence. 15. Report ' A report containing all the results o f the stndy will be prepared by the 3M Environmental Laboratory. If analyses are sub-contracted to other laboratories, each laboratory will prepare a report and submit it to the 3M Environmental Laboratory for inclusion in the 3M Environmental Laboratory report. F-1^ report will include, but not be limited to, the following, when applicable: . '` . 15.1 Name and address o f the facility performing the study " 15.2 Dates upon which the study was initiated and completed 15.3 A statement ofcompliance by the Study Director addressing any exceptions to Good Laboratory Practice Standards 15.4 Objectives and procedures as stated in the approved protocol, including any changes in the original protocol '- 15.5 The test substance identification by name, chemical abstracts number or code number, strength, purity, and composition or other appropriate characteristics, if provided by the Spbnsor ' 15.6 Slability and the solubility of the test substances underthe conditions of administration, if provided by the Sponsor . ., . 15.7 A description of the methods used to conduct the test(s) ' 15.3, A description ofthe test system. . 15.9 A description of any circumstances that may have afiected the Quality or the integrity o f the data . 1 5 .1 0 The name of the Study Director and the names ofotherscientists, professionals, and supervisorypenonnel involved in the study . 15.11 A description of the transformations, calculations, or operations performed on the data, a summary and analysis of the analytical chemistry data, and a statement o f the conclusions drawn from the analyses 15.12 Statistical methods used to evaluate the data, if applicable 15.13 The signed and dated reports of each'ofthe individual scientists or other professionals involved in the study, if applicable 15.14 The location where raw data and the final report are to be stored 3M E n v iro n m en ta l Laboratory Centre Analytical Laboratories, Inc. 3M Environmental Laboratory P a g e a o f 10 Page'57 of 103 Page 305 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Protocol "FACT-TOX-110-. 15.15 A statement prsred by the Quality A ssum es Unit listing the dates tha s!U(jv inspections and audits were made, and the dates o f any findings reported to rh' Director and Management1 e ay U it is necessary to make corrections dr additions to a report titer it has been accented, th caanges will be made in the form of an amendment issued by the Study Director. The * neadment will dearly identify the part of the report that is being amended, the reason* f,, .t. ttesdment, and will be signed by the Study Director. re 1 6 . L o c a t io n o f R a w Da t a , R e c o r d s , a n d Fin a l R e p o r t 9 n.s!nai ot copies thereof; win be available at the 3M Environmental laboratory to crhtate audits ofthe studydaring itsprogress and before'accsptance o f the final resort Wfcm ute final re art iscomp!eted,-an originai paper data, including hose items !r*t~ f{,ejQW ^ . ream ed i n . :a archives of3MEnvironmental Laboratory for at least a period o f time as cn j ' by regulatic -, and as established by 3M Environmental Laboratory Standard Operating TM*Ciae >o. 7 i l e -allowing raw data and records will be retained i Proc lares: crating 16.1 Approvedprotocol and amendments `16,1. -Stady correspondence 16.1. Shippingrecords 16.1. Haw data 16.1.. Approved final report (original signed copy) 16.1.5 ecironic copies o f data 15.2 The foL ing supporting records will be retainedseparately from.the study folder in the arch: : according to 3MEnviroameatal.Laboratory Standard Operation Procedure * S 16.2.1 7 iningrecords 16.2.2 C hratiorecords 16.2.3 ihsr meatmaintenance logs 16.2.4 Standard Operating Procedures, Equipment Procedures, and Methods 3 M E n vironm ental Laboratory Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Pagi 9 of io . Page 58 o f 103 Page 306 3 M Med.i.cal _Depart.ment. Stud.y. TT-c6o2n9e5.t1o2 i Analyytical Report: FAC. T.T. O. jX. .-110 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 ' . P.-cfcce/ & A C 7 -T C X -1 1 0 17. SPECIMENRETENTION . Specimens will be maintained in the SMEnvironmental Laboratory specimen archiv--for period of time as specified by regulation or as long as the quality of the preparation affords evaluation, but not longer than ten years following the effective date ofthe final test mle flf applicable), and as established by 3M Environmental Laboratory Standard Operating*"roc-dures 1 8 . P r o to c c lA m e n d n e n t sa n d d e v ia t io n s asPlanned changes to the protocol will be in the form of written amendments signed bv the Stu- Director and the Sponsor's Representative. Amendments will be considered part of the- protocol and will be attached to the final protocol All changes to the protocol will be indicated m the final report. Any otherchanges will be in the form o f written deviations sianed hv tk Study Director and filed with the raw data-. '- y e - 1 9 'A ttac h m en ts . . 19.1 Attachment A Preparatory and analytical methods 2 0 . S ig n atu res Manon T. Case, D.VJvL, PhD., Sponsor Representative QA 0 Date ' ML V - L - . ____________ ttnsien J. Hansen, PhD., 3M Environmental Laboratory Study Director Date SM Environm ental Laboratory Centre Analytical Laboratories, Inc. 3M Environmental Laboratory . PagalOcflO Page 59 of 103 Page 307 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Study Titie Oral (Gavage) Pharmacokinetic Study ofPFOS in Rats PROTOCOL AMENDMENT N O .1 Amendment Date: August 12,1999 . Performing Laboratory 3M Environmental Technology & Safety Services 3M Environmental Laboratory 935 Bush Avenue * S t Paul, MN 55105 Laboratory Project Identificatv n ET&SS FACT-TOX110 URNU2S49 EcsctCopy of Original 3M Environmental Laboratory Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 60 of 103 Page 308 3M Medical Department Study: T-6295.12 , Ana|ytical RePort: FACT TOqX' 110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 ., ' ', Frotcccl FAC 7-TO X110 ' Amendment No. 1 Tnis am endm ent m odifies the following porticn(s) of the protocol: 1. P r o t o c o l r e a d s : Section 2.0 states this study is designed to determine potassium perfloorooctane sulfonic acid (PFOS) in specimens of liver and serum of rats. Amend TOread: This study is designed to determine PFOS in specimens ofrat liver, serum and urine. AHDay -1 and Gestation Day 21 rat urine specimens will be extracted and*analyzed by. the 3MEnvironmental Laboratory. R e a s o n : The urine extraction and analytical methods were not validated and approved prior to protocol approval. . * 2. P r o t o c o l r e a d s : S e c tion 6.0 lists rat or rabbit liver and serum. A m e n d t o r e a d : Sat or rabbit urine from 3M Toxicologywith a physical description o f rat or rabbit urine. R e a s o n : The iat urine matrix was added after the protocol was approved. 3. PROTOCOLREADS: Section. 10.0 and 11.0 list the Mowing methods to use for extraction and analysis: 1. FACT-M-1.1 "Extracdoa of Potassium Perfluorooctanesulfonatc or Other Anionic Fluorochemical Surfactant from Liver for Analysis Using HPLC-Electrospray/Mass Spectrometry" '' , FACT-M-2.1 "Analysis o fFluorocaemicals in Liver Extracts Using HPLC-Electrospray/Mass Spectrometry" '" A m e n d to read: The extraction and analytical methods to fallow at the 3M Environmental Laboratory are: ' ETS-8- .o "Extraction o fPotassium Perfluorooctanesulfinate or 'OtherFluorochemical Compounds from Liver for Analysis Using HPLC-Elcctrospray/Mass Spectrometry" ETS-8-96.0 "Extraction o fPotassium Perfhiorooctanesulfonate or Other Fluorochemical Compounds from Urine for Analysis Using HPLC-Electrospray/Mass Spectrometry/Mass Spectrometry" ETS-8-7.0 "Analysis of Potassium Perfluorooctanesulfbnaie or OtherFluorochemical Compounds in LiverExtracts Using HPLC-Electrospray/Mass Spectrometry" ETS-3-97.0 "AnalysisofPotassium Perfluorooctanesulfonatc or Cther Fluorochemical Compounds in Urine Extracts Using HPLC-Electrospray/Mass Spectrometry/Mass Spectrometry" 3JWEnvironmentalLaboratory Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 61 of 103 Page 309 Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 FrctocdrACT-TOX110 AmendmentNo. 1 Reason: The extraction and analytical methods FACT-M-1.1 and FACT-M-2.1, recce-tive'v were updated on 07/22/99 to ETS-S-6.0 and ETS-8-7.0. ' ~ `y ` Methods ETS-3-96.0 and ETS-3-97.0 were not validated and approved until after the -'rotoeil was approved. ' "" 0 4. PROTOCOL r e a d s : Section 10 .4 and 11.4 state that i f the analyses are sub-contracted to other laboratories an amendment will he written to include these methods. Amend toread: The extraction and analytical methods to M ow at Advanced Bioanalytical ' Serviceswinbeattachedtotheprotocol- . The extraction and analytical method to follow at Battelle Memorial Institute is: "Method for Analysis o f Perflnotooctane Siilfonate (PFOS) in Sat Seraby LC/MS/MS Version V i 0 Bw ' REASON: The analytical methods at the sub-contract laboratories were not included in foe original p r o to c o l 5. Protocol reads: Section 12.2.1 b) Liver method detection limit is 15 ppb. Amend to read: 12.2.1 b) Liver method detection limit is 3.50 ppb (ng/g). 12^.1 c) Urine method detection limit is 1.5 ppb (ng/g). Reason: The validation supporting methods ETS-8-6.0 and ETS-8-7.0 includeda lower method detection limit for PFOS in liver. A validation in urine was perforated, after protocol approval, to support fois method detecaon limit Ijj, Ite rili 3M environmentalLaboratory ' Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 62 of 103 Page 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 . Amendment Approval - Prztccd FACT-TQ Xno Sentiment iVq. 1 ------------- f y f l a/w^-- Marvin Case, D.V2&, PhJ).. Sponsor stanve _/7 ,v7- fag Das & '.... J. Bansea, PhJ3,, Study Director Date Environmental Laboratory Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 63 of 103 Page 311 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 . Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 ' Study Title Oral (Gavage) Pharmacokinetic Study ofPFOS in Rats PROTOCOL AMENDMENTNO. 2 Amendment Date: September 23,1999 . Performing Laboratory 3M EnvironmentaTeehnoIogy & Safety Services 3M Environmental Laboratory 935 Bush Avenue S t Paul, MN 55105 Laboratory Project identification ET&SS FACT-TOXI10 LIENU2S49 3WE nvironm ental Laboratory ; Exact Copyof Original UC. ' initiai (tllU jo Oats Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 64 of 103 Page 312 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 . LRN-U2849 , Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 ' _ . ' fote&IFACT-TOXflO : ' Amendment No. 2 This amendment modifies the following portfcn(s) of the protocol- 1- P r o t o c o l r e a d s : Section 2 states that all liver s a m e lts will be extracted and analyzed at Battelle Memorial Institute. ' 'w A m end to r ea d : . All dam fiver specimens (female mother) and 27 fetus (pooled) fiver specimens will be analyzed at Battefie Memorial Institute. Reason: The fetal livesflimg is not a targetmatrix and will not be extracted or analyzed. -2 . P r o t o c o l r e a d s : Section 9 states that 80 dam liver specimens (female mother) and 80 fetus (pooled) fiver specimens will he seat to the Environmental Laboratory. ` R eason: The number of an im als changed during the course of the study. 3. P r o t o c o l r e a d s : Section 12.2.1 a) serum method detection limit is 1.75 ppb b) liv er method detection limit js 15 ppb. ' . I Amend to read: The method detection limits for all compounds and matrices will be taken fiom the methods Used for extraction and analysis. R eason: Tae method detection limits fisted are specific to the 3M Environmental Laboratory. Stateateat was added to aliow for sub-contracted analyses and/or revised methods. 3M Environmental Laboratory Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 65 of 103 Page 313 3 M Medical Department Study: T-6295.12 ; Analytical Report: FACT TOX-110 LRN-U2849 , Centre Study No.: 023-016 SponsorProtocol No: FACT-TOX-110 . : pratccdFACT-7OX1l0 ' An&xhnentNo. 2 4. P r o t o c o l r e a d s : Secdon 16 states that the original data, or copies thereof, will be available at the 3M Environmental Laboratory to facilitate audits of the study during its progress and before acceptance of the anal report When the final report is completed, all original pacer data, including: approved protocol and amendments, study correspondence, shipping records raw data, approved final report, electronic copies o fdata, training records, raffire.-,, msinnneat maintenance logs and standard operating procedures, equipment procedures mid methods will be retained in the archives o f the 3M Environmental Laboratory. . ' A h e m to read: . Section 16 states that the original data, or copies thereof 'vill be available at the 3M Environmental Laboratory to facilitate audits of the study during its'progress and before acceptance ofthe final resort When the final report is completed, all oriufnaT j --. eroding: approved protocol and amendments, study correspondence, shipping raw data, approved final report, andelectronic copies o f data will be retained in the archives of the 3MEnvironmental Laboratory. All corresponding training records, ealibrari^ reCQr<is matrumeat maintenance logs, standard operating procedures, equipment procedures and methods will be retained in the archives of the facility performing each analysis. ' R e aso n : .. Clarification ofthe disposition ofarchived records if analyses are performed at a sub-contract laboratory. '_ - - 5. Protocol read s: ' Section 17 states that specimens will be maintained in the 3M Environmental Laboratory specimen archives. A mend to read : ' S p e c im e n s w ill b e m a in ta in ed in the3M Environmental Laboratory s p e c im e n a rc h iv e s. A l l specuncus sent to sub-contract laboratories will be returned to the 3M Environmental Laboratory upon completionofanalysts and submission o f th e s o b -c o n tra c t labonttory(s) final report. The specimens will be returned with the following documentation: the signed original chain of custody andrecords ofstorage conditions while at the sub-contract facility. R eason: ` performed by a sub-contract laboratory. 3JWEnvironm ental Laboratory Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 66 of 103 Page 314 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 . Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Amendment Approval PrctcczIFACT-TCXUO A m en d m en t Vo. 2 Marvin Case, D.VAL, PhD., Sponsor Representative K rien J. Hansen, P h D , Study Director lQ4 ' Date . 3 M Environm ental Laboratory Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 67 of 103 Page 315 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 , Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 v. Study Title . Analytical Laboratory Report on the Detennination ofPeriluorooctanesulfonate (PROS) Presence and Concentration in Serum, Liver, and Urine from the (ravage Study ofT^6295 P PROTOCOL AMENDMENT NO. 3 Amendment Date: 20 January 2000 ' Performing Laboratories Urine Analyses Liver Analyses 3M Environmental Technology and Safer/Services Batteile Memocal Institute F lu o r ic Analytical Chemistry T o m 505 King Avenue Building 2-3E-09 Columbus, OH 4JZQ1-2693 935 Bush Avenue St. Paul, MN 5510 ' . Serum Analyses Advanced Bloanaiytieal Services Inc. ' 15 Casherwoad Road Ithaca, N Y 14S0 Laboratory Project identification ET&SS LRN-U2S49 FACT TOX-110 Argus Study: 41S-0I3 . 3M Medical Department Study: T-6295.12 3M Environmental Laboratory Esc; Copy0f original - M * --- Siii b Initial Oats ' Centre Analytical Laboratories, Inc. . 3M Environmental Laboratory . Page 68 of 103 . Fage 316 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 , Centre Study No.: 023-016 ' Sponsor Protocol No: FACT-TOX-110 . .' ` Proteccl LF.N-U234S Amendment Num cer 3 This am endm ent m odifies the folowing porton(s) of the protocol- 1- P r o t o c o l r e a d s ; The study director for the present study w as identified in the protocol as Kris*en j Hansen, Ph.D. `` ' A mend to read : . Therole o f study director for the present stu d y w a s r e a ssig n ed to M arvin T C a sa M M , Ph.D., a s of 20 January 2000. The previous study director, Kristen j ' H ^ se n , has been reassigned to the role of Principle Analytical investigator' The role of study director w as reassigned in an effort to ensure compliance with ' Good Laboratory Practice Standards that outline study personnel reouimmB,,i,, (refer to 21 CFRParf5S). . nts 2. P r o t o c o l r e a d s : , The sponsor for the present study w as identified as Marvin T. C ase n v u :A m e n d t o r e a d > . rn.u. The role ofsponsorforthe present study w as reassigned to John L Ph.D., a s of 20 January 2000. ' R eason: . . . ' To ensure that toe study director does not also cany toe duties of study sponsor the sponsor role w as reassigned. In this manner, personnel responsibilities and ' ' workload are more evenly balanced. ' 3M Environments/Laboratory Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page'69 o f 103 . Page 317 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Amendment Approval f r e c c i LPM-U2S49 AMflfitmant Number 3 John L. Butenhoff, PhD., SponsorReprcsentarive Kristen J, Hamen, PhD., Outgoing Study Director Mr --?;/> Date 2M E nvironm ental Laboratory Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 70 of 103. Page 318 3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 . Centre Study No.: 023-016 ' Sponsor Protocol No: FACT-TOX-110 . S tu d y Title Orai (Gavage) Pharmacokinetic Study ofPFOS in Rats PROTOCOL AMENDMENT NO. 4 A m endm ent Date: 20 April 2000 Performing Laboratories 3M Environmental Technology and Safety Service! Fluorine Analytical Chemisoy Team Building 2-3E-09,93J Bush Avenue: S t Paul, MN 35106 ' Centre Analytical L aboratories, TM . - 304S Research D rive' State OiHege, PA 16301 - Batteiie Memorial Institme . 305 King Avenue Columbus, OH 43201-2693 Advanced Bloaaalytieai Services, 5 , t 13 Catherwood Road rthaea, NY 14S30 Laboratory Project Identification ET&SS LRN-U2S49 FACTTOX-IIO Argus Study: 418-013 3M Medical Department Study: T-629.12 3M Environmental Labom tor/ Centre Analytical Laboratories, Inc. . 3M Environmental Laboratory . Page 71 of 103 Page 319 3M Medical Department Study: T-6295.12 1 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 " ' Sponsor Protocol No: FACT-TOX-110 . ': P r e te x t LF.N-UZZ49 . Amendment Num ber 4 This amendment modifies the following portions) of the protocol: 1- Protocol reads: The amended section 2.0 text states that this study is designed to determine PFOS in specimens ofrat liver, serum, and urine. . Amend to read: 1 ` .. This study is designed to determine PFOS in specimens ofrat liver, serum, feces, and urine. Reason: . . ' The analysis offecal tissue forthe target chemical and/or its analytes was added to the scooe o f the study following the issuance of the protocol Feces extraction and analytical methods were not validated and approved prior to protocol approval ., 2. Protocolreads: The amended section 6.0 lists rat orrahbit liver, serum, and urine. Amend toread: Add: rat or rabbit feces with a physical description ofrat orrabbit feces. R eason: . Analysis o f fecal tissue for the target chemical and/or its analytes was added to the scope o f the study following the issuance of the original protocol. 3- Protocol-Reads: - * Section 13.1 lists ail of the laboratories that will be conducting analyses for this study. A mend toread: ' Add: Centre Analytical Laboratories, Inc., 3048 Research Drive, State College, PA 16801 Reason: .. Feces analyses were added to the scope of this study. The sub-contract laboratory performing analyses was not in the original protocol. = 4. Protocol Reads: Sections 10.4 and 11.4 stale that if the analyses are sub-conlracted to other laboratories an ' amendment will be written to include these methods. . Amend to read: . The feces extraction and analytical method used by Centre Analytical Laboratories will be* OOM-023-003 (Revision 2), "Determination of Fluorochemical Residues in Monkey/Rac ' Feces by LOMS/MS." R easo n : The sub-contract laboratory performing feces analyses was added to the scope o f this study; this method was not validated and approved prior to protocol approvaL ' 3MEivvnrnentalLaboratory Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 72 of 103 Page 320 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 f a t c c s l LRN-U2 B4 S Amendment N um ber 4' Amendmenf Approve! ohn L Buienhojj, PAD., Sponsor Representative C u .* ; ____________ Marmi T. Crue, D. VM,, PhSt,, Study Director 3M Environmental Laboratory Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 73 of 103; Page 321 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-llO 3043 Research Drive, State College, PA X680L Pbooe: (SM) 231*8032, Facsimile: (814)231-1253 3Tl,d/ bt/eertr Centre Analytical Laboratories, Ine. 3M Environmental Laboratory bar-i. Page 74 o f 103 Page 322 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849' . Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 3048 Research Drive, State CoUeg* PA 16801* Phoae: (814) 231-8032, Facsimile} (314)231-1353 PROTOCOL DEVIATION Deviation Number 2 , . Date of Occurrence: (1) entire study Centre Study Number: 023-016 Centre Protocol Number. FACT-TOX- 1 10 ' DESCRIPTIONOF DEVIATION .? 1. Amendment 2 Item A method detection limit was not established forrat feces during the method validation. nwfa)_______________________________ ________ ACTIONS TAKEN i.e.. amendment issued. SOPrevirion, etc- 1. Protocol deviation issued,^--s. Recorded By/Date: B hjm IMPACT ONTHE STUDY 1. No negative impact. RATIONALE I. Any sample that contained residue with - 3:1 signahnoise was integrated and any number that was greater than 0 was reported. The limit of quaniration was established at 10ppb during the method validation. e ftincipal Investigator Signature k TQhul- SponsorRepresentative Signature CAL QAUReview. F / f f a -o Date A-ut Date * : f'u February 12. 1998/2 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 75 of 103 Page 323 3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 APPENDIX B Determination of Fluorochemical Residues in Monkey/Rat Feces by LC/MS/MS (Revision 2) Method #00M-023-003, revision 2 and Deviations and Modifications Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 76 of 103 Page 324 3rd Medical Department Study: T-6295.12 , Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 TITLE Determinanon of Fluoroehemicai Residues in Monfcey/Rat Feces bv LC/MS 'Me (Revision 2) ' ' n^ AUTHORS Bnnksha Wickremesinhe, ShaazhiZheng, andJohnFlaherty DATE ISSUED June 02,2000 SPONSOR 3M Environmental Technology and Safety Services' Building 2-3E-09 - PO Box 33331 SL Paul, MN 55133-3331 PERFORMING LABORATORY Centre Analytical Laboratories, Lie. (Centre) 3048 ResearchDrive State College, PA 16801 ' ' CENTRE STUDYNUMBER 023-003 CENTRE METHOD NUMBER 0OM-O23-O3, revision 2 TOTAL NUMBER OF PAGES 20 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 77 of 103 Page 325 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 . C san e M n hod N o. ; OOM-02J-003, revision 2 MANAGEMENT APPROVAL The work cited here was performed in conformance with applicable standard o *>' procedures and general GLP's. pe.<-nng Principal Investigator . . Centre Analytical Laboratories, Inc -O Z-O O _________p__L__--___T_______ - C >2 -e n D Shaozhi Zheng ' Date Principal Investigator Centre Analytical Laboratories, Inc. Laboratory Manager Centre Analytical Laboratories, Inc. 2 -UwvV-io f iT -fr/ (> OA i-MtNi- L Kris Hansen, P tD . Group Leader 3M Environmental Laboratory /O 0/Q Date C intre Analytical Laboratories, lac. Study # 023-003 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 2 Page 78 o f 103 Page 326 Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 C esflcM ehod No. : 0OM-O22-CO3, revioa 2 TABLE OF CONTENTS TITLE PAGE--------- -- -------- I MANAGEMENT APPROVAL--------------------------------------------------------------- 2 TABLE OF CONTENTS-- ----------------------------- 3 1. SUMMARY__________;-- ------------------------------------------ --------------- 4 2. FLUOROCHEM3CAL STANDARDS........ ................ 4 3. CHEMICALS AND SUPPLIES------------------- -------------------------------------- - 3 .1 . Ch em ica ls------------- ------------;---------------------------------------------------- ---------- 7 33. FluorochemicalStandares-------------- ----------------------- !------------- 7 3 3 . EquipmentandSupplies-- ------------------------ --- ------------------------- - 3.4. Solutions----------i-- -- ---------------------- ------------------------------ s 3.5. PreparationofStock, Foruhcation, and Calibration Solutions....... 9 3.5.1. Stock solutions-------------- y-- -------------------------------------- - 3 3 3 . Fortification Solutions----------------------------------- ,---------------- 9 3 3 3 . Calibration Standards---------------------- --- ------------------------- - 4. METHOD______________________________________________ _ 4 .1 . Flo w Diagram -- ----------------------'---------------------------------------------- --------u 4.2. SampleProcessing------------------------ -------------------------------------- 43. SamplePreparation--------------- ------------------------ -------------- n 43.1. MatrixBlanks------ .---------------------------------------------------- 1 2 . 4 3 3 . Mat: Zero Blanks--------------------------------------------- -- ...1 2 4 3 3 . CalibrationStandards-- ---- ----------------------------------------- 43.4. Continuing Calibration Verification Standards------------------------- - 4 3 3 . Q C Recovery Samples-- ---------------- ------------------------ :..12 4.4. Extraction------------------------------------------ -----...........-- 12 4.4.1 SPE ColumnPreparation------------------------------------------------- 4.43 SDanizationofFear-SbapedFlasks----- -------------------------------- 13 4.43 Standardization ofSPE Columns--------------------------- ,, ..1 3 4 3 . A nalysis by LC/MS/MS -- ------------------------------------1---------------- 14 4 3 .1 . LC/MS/MSsystem and Operating Conditions (Electxospray)_____ 14 433. ExampleTuneF3eParameters-------------------------------------- - 433. CalibrationProcedures----------------------------------------------------16 43.4. SampleAnalysis----------------------------------------------- 17 4.6. PerformanceCriteria-- -- ------------- ;-----------------------------------i s 4 .7 . T im e Required for Analyse------------------------------ -- --------------- -----------13 5. CALCULATIONS--------------------------------------------------------------------- - 5.1.--- ---------------------------------------------------------------- IS 5 3 . Q C R eco v ery -- - -- --------- --- --------------------------------------- ---------- .1 9 5 3 . Me / ResponseFactor----- ---------------------------------------------------- - 6. SAFETY__________________________________________________ _ ATTACHMENT ________________________________________ 30 CenereAnalytical Labcraroria, IncStudy #022-003 Page 3 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 79 of 103 Page 327 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849-- Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-l 10 1. SUMMARY C eaae M e th o d H a. : OOM.023-003, revision' 2 Unis document details a method of analysis for residues of perfluoroocune sulfonate (PFOS), perfluorooctane sulfonylamide (PFOSA), perfluorooctane suifonyiamiaoethyi- acetate (PFOSAA), perfluorooctmesulfonylethylamide (PFOSEA), perfluorooctanoate (POAA), 2(N-ethy!pen3uorooctanesulfonanudo}rethy! alcohol (Et-FOSE-OK), M570 and MJ56 in monkey feces. The chemical formulas of the analytes are given in Section v of this method. ." This method is being re-issued as method 00M-023-0Q3 revision 2. It is being revised to add the rat feces as a matrix,.to document the use of20-mL vials instead 0f 5 ^ ^ for sample extraction, to add the option of homogenizing tests samples before weighin'* to clarify the preparation of QC recovery samples and to correct some typographical tots in method OOM-023-003 revision 1 . Rat feces is being added to the method, however; it was validated for the analysis o f PFOS only. Therefore, this method sh o u ld be used for the analysis ofPFOS only, in rat feces. ' Residues of these ihiorochemicais are extracted from feces with acetonitrile (ACN) The ACN extract is filtered and passed to n g a a carbon solid-phase extraction (SPE) column. Approximately 3-4 drags o f l-octanol are added to the extract and evaporated to near thyness using rotary evaporation. Each extract is then reconstituted-'with methanol Quantification ofPFOS, PFOSA PFOSAA, PFOSEA,- Et-FOSE-OH, M356, M570 and POAA is accomplished by liquid ehrornatography/tandem mass spectrometry (LC/MS/MS) analysts using selected reaction momtoring (SRM). * The proposed limit of quantitation (LOQ) for this method is 10 ppb each for PFOS PFOSA, PFOSAA, PFOSEA, Et-FOSE-OH, U 5S 6 , M 570, and POAA. This is based on studies conducted during the development of this method using control monkey feces 2. FLUOROCHEMICAL STANDARDS Molecular structures of PFOS. PFOSA, PFOSAA, PFOSEA, Et-FOSE-OH, M556 M570, and POAA are given below. ' PFOS Molecular weight 499 (Q F^O j- ) 0 II c <-tjS-- Or M CseaicalNarae Pertliiorooenne sulfotutt . Note: The neutral molecule and standard form that the.PFOS (anion) is derived from is potassium perfluorooctanesulfonate (C,F17SOjK], molecular weight S38 CeaseAnalyticalLaboratories, Inc. Study*023-003 Page 4 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 80 o f 103 Page 328 3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 .' ' ' M ethod >io. ; OOM-023-003, rev isio n s PFOSA Molecularweight 4S9 0 ' C9F17S-- NH2 0 Chemical Name = Periluorooctane sulfonylamide ' ' PFSAA . Moleealarweight585 . ' I ^OVCOH C(r]7S--- H I! ' `. Chemical Name = periluoreoctane suifonylamidoetltylaeetate ' 1 ' PFOSEA Molecularweight527 . . C*H71S-- N/ " 0II . . Chemical Name Perduoroocsnesulfbnylethylaraide POAAMolecular weight 413 C7F 15CCOChemical Name * PerHuoroomanoate Note:TheneutralmoleculeandstandardformthatthePOAA(anion)isd ` d from is ammoniumperfluoroctanoate [C^jCOONHJ, molecular weight 431 C anne Analytical Laboratories, Inc. Study 023-003 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory PageJ Page 81 of 103 Page 329 3M Medical Department Study. T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Method No. : 0M-0 03, revisten 2 Et-FOSE-OH Molecularweighc 571 Il / OtftCH CjrirS---N O OVH. Chcaical Name - 2(N-etliyIperiIaorooimmesuIfonaniido)-e!liy! aleVcel VI556 . Molecularweighc 557 I ^CHfiOOH Cjr^S-- N 0II Q eazical Name **M 556 M570 Molecularweighc 571 jj yCffeCOCH C=,7S-- N oII '''C H j fV~tieal Name M570 TfflPFOS Molecularweight 423 I-H, 1-H, 2-H, 2-H, CgF^SOjH Ctecal Name -4-H,perfIuorooctane sulfonic acid Ceaire Analytical Laboratories, l o t Study 023-003 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 6 Page 82 of 103 Page 330 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 CentreM e io d N o. : D 0M 2M 03, revision 2 3. CHEMICALS ANDSUPPLIES I' ' 3.1. Chemicals Chemical Acsiomtriie (ACN) Carbon 120/400 mesh Methanol (MeOH) 1-Octanol Ammonium Acetate L-Ascorbic add Toluene Dimetyldichlorosilane Water Grade Source HPLC - EPLC Ragent Ragent Reagent Reagent Reagent Type I (VWR)J.T. Baker . Supelco (VWR) J. T. Baker Aldrich Chemical Sigma-Aldrich Signia-Aldrich . (VWR.) L T. Baker , Supelco in house Catalog N0. ~ JTS0L7-3 ~ ' 57210-U JT3093-2 111-37-5 A-7330 L-5960 JT9460-3 3-3009 (Type I water= electrical resistivity, minimum'of 16.67 MO/cm at 25"C from Lahconco watepro workstation) ' 3.2. F luorochemical Standards Standard PROS PROSA PROSAA PROSEA Et-ROSE-OH M5J6 M570 POAA ` THPEOS . 1 . Source 3M 3M 3M 3M 3M 3M 3M 3M 3M 3 .3 . Equipment and Supplies __________Equipment________ Balance, analytical, (display at least 0.0001g) Balance, top loading, (display at least 0.01 g) Rotary evaporator Rotary evaporator trap Pear-shaped flasks (125 mL) 20-mL SPE tubes (cat. no. N057177) with frits Vacuum pump . - Visiprep vacuum manifold . Ceatre Analytical Laboratories, Inc. Slady * 023-003 Source Mettler Mettler Buchi Buchi Pyrex Supelco Buchi Supelco Page 7 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 83 of 103 Page 331 3M Medical Department Study. T-6295.12 Analytical Report: FACT TOX-110 . LRN-U2849 ' Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 C caw M ebodW o.: OOM-OZjx , , : ^ ^ , Wrist-action shaker ' 20-mL polyethylene scindlladon vials with lids 20-mL syringe ' 25-mm diameter glass fibre acrodise (cat # 4523) Disposable pipets, test tubes etc. Disposable micropipets, 100- 200 iL. 125-mL LDPE narrow-mouth bottles . 2-mL clear HPLC vial kit (cat #5181-3400) Standard lab equipment (classA pipets and volumetric flask, graduated cylinders, etc.) LC/MS/MS and HPLC systems*124 Burrell Wbeaon (VWR) vwa. Ge'anan . VTVR Nalgene f5> various Asdescribedin Section4.5.T Li order to avoid contamination, the use of disoosable containers/tubes^ipets etc. is highly recommended. ' Teflon or teflon-lined containers or equipment, ncludina teflon-hned HPLC vial caps should not be used. 5 Pear-shaped flasks are silanized before use. It may be necessary to check the solvents (methanol) for &- presence of contaminants (especially POAA) by LOMS/MS before use. Certain lot numbers have been found to be unsuitable for use. ' Disposable micropipets or pipets should be used to aliquot standard solutions, when preparing standards and samples for extraction. _ Equivalent materials may be substituted.for those soecified in this method. However, the use of carbon fiom Supelco is strongly recommended. y 3.4. Solutions 1. 50 mM Ammonium Acetate: Dissolve 3.85 g of ammonium acetate in 1 L of typeI water. 2. 2 mM Ammonium Acetate: Dilute 40 mL of the 100 mM acetate sohrdon in a liter of type1 water, for mobile phase A. . ' 3. SaturatedAscorbicAcidinMelbanol:Dissolve-10 g ascorbic acid i 100 mL methanol 4. The 2% Ascorbic Add in Methanol: Dissolve 2 g ascorbic add in 100 mL methanol Note: The volumes shown are provided for guidance; alternative v o ln m may be prepared. . Crre A nalytical Laboratories: Inc. Study if 023-003 Page3 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 84 of 103 Page 332 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-0 L6 Sponsor Protocol No: FACT-TOX-110 C zsa c Method No. : OOM-OLMaj, revision 2 3.5. PREPARATION OF STOCK, FORTTKCATTON, AND CaUBRATTOSI Saum ons Analytical standards are used for two purposes: (l)Jo fortify consol matrix samples designated to be extracted and processed as calibmnon a a n T ^ k l T ^ n be then used to calibrate the response o f the detector used in the analysis:' -won to fortify other control matrix samples to determine analytical re c o v e ry ' ^ ~ The absolute volumes of the standards may be varied by the analyst as 1 correct proportions ofsolute to solvent are maintained. The solutions cited iT i * ' are given as an example; alternative concentrations maybe prepared ifneedeti W To prepare stock solutions of 100 pg/mL each PFOS, PFOSA P u n e * PFOSEA, Et-FOSE-OH, M556, M570, POAA, and the surro^te s r a n ^ J IHPFOS, weigh out 10 mg o f analytical standard (corrected fm n u S ^ percent salt, if necessary) gad dilute to 100 wL with methanol m a lflL r volumetric flask. Prepare a separate solution for each analyte Th ^ u solutions Cm125-mL LDPE bottles) are to be stored in a refrigerator at 2 r t 6C and are stable for a maximum period of 6 months from the ' t # 'preparation. ' oate of 3.5.2. Fortification Solutions ' a. 10 pg/mL Mixed Fortification Solution - Pipet 10.0 mL each nr w n c PFOSA, PFOSAA, PFOSEA, Et-FOSE-OH, M556, M570 and PQa a stock solution to a 100 mL volumetric flask. Bring up to volume with methanoL * wun b. -2.S usfaLMixedFortifiC3tionSolucon-Pipet25.n mr jp , - mixed fortification solution to a 100-mL volumetric flask and briit ^ volume withmethanoL S^pto c. 0 J pg/mL Mixed Fortification Solution - Pipet 20.0 mL o f the 25 at r .mixed fottification solution to a 100-mL volumetric flask and brin volume with methanoL ' 5 upto d. 0.1 pgtoLMixedFortificationSolution-Pipet20.0.mLoftheOJug/mL mixed fortification solution to a 100-mL volumetric flask and brin*1* 13^ volume withmethanoL S up to Follow Steps a and b above to make a 2.5 pg/mL solution o f the su standard (IHPFOS) from the stock solution. *Sate C aa e A nalytical Laboratories, loe. Study *023-003 Page 9 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 85 of 103 Page 333 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 ' C o a t M ethod N o .: OOM.023-C03. - v i n c a 2 Store all fortification standard solutions (In 125-mL LDP- fc0r '- i - refrigerator at 2C to 68C for a m n im ait period of 6 months from th ^dat-of preparation. ' -t 3.5.3. Calibration Standards , Prepare LOMS/MS calToration standards by fortifying weighed out alof the same control feces sample (or a combined '"bulk" control feces before they are processed through the Retractionprocedure. * Sam The following is a typical example; additional concentrations may be D renared as needed. It is recommended to prepare the fortification solutions so that the volume of fortification is between 50 and 200 pL and to use disn h i' micropipets for iiiquotmg fortification volumes. ` 0sai) C Cone, of Fortification Weight of Mixed Volume (jL) Control Fortification Sample (g) Solution ()ig/ml) NA NA 1.0 0.1 0.1 0.5 0.5 100 200 100 200 1.0 1.0 * 1.0 1.0 2.5 2.5 10 100 ' 200 100 1.0 1.0 . 1.0 9 Zo pg/ml THrrOS fortification solution. Cone, of Extracted Calibration Standard (ppb) NA 10 20 50 100 250 500 1000 VoL of Surrogate Standard* added (pL) 2 QQ 200 200 200 200 200 ' 200 200 wuoranon scmaaiu prejjmiuasw--- - * reixjgerator at 2C to 6C, if needed, and used over a period of time as long as its stability has been verified. . . " Csatre Analytical Labenioria, Inc. Study 9023-003 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 10 Page 86 of 103 Page 334 3M Medical Department Study. T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre StudyNo.: 023-016 Sponsor Protocol No: FACT-TOX-110 . CramMeihodNo.: QOM-OZS-Mtjrevirion, 4. METHOD 4.1. Flow D iagram : r . T ie flow diagram o f the method is given below, followed bv a ' , description of each step. ' ' * Qelued ' Method Flow Diagram Weigh 1.0 g ofsample ' (fortify samples designated as calibration standards and QC recoye-es) . . -' ' 4-' Extract with ACN and filter 4. . .4 Carbon Solid-Phase Extraction l * Concentrate to almost dryness i i' Adjust final volume (2.0 mL) 4 4. 7-OMS/MS ' . * 4.2. Sam ple P rocessing . All samples are received frozen and will be kept frozen (below - 10Q ,,-t of extraction. The rat feces do not need any processing, however, the monk * feces may need to be homogenized in order to be able to weigh out'th- SDec'^d amounts. " . Jiled 4.3. Sam ple Preparation Prepare the following blank and calibration samples (Sections 4 .3 .1 to 4 " <>r. same control feces sample (or a combined "bulk" control feces sample) ~ ra Cesire Analytical Laboratori, Inc. Study# 023-003 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page It Page 87 of 103 Page 335 3M Medical Department Study: T-6295.12 i . Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.; 023-016 SponsorProtocol No: FACT-TOX-110 C a m Method N o.: OOM-O2j-O03,~vision 43.1. M atrix Blanks . Prepare two blank samples without any fortifications, per set of samles analyzed. These will be.ttsed as the matrix blank. " 4.3.2. M atrix Zero Blanks ` Prepare two matrix blank samples fortified with the surrogate, per set o f samples analyzed. These will be used as the matrix zero blanks. ' 4.33. Calibration Standards Prepare calibration standards as described in Section 3.5J ; 43.4. Continuing Calibration Verification Standards Two of the extracted calibration standards, a low-mid and a mid-high level standard, will be used as continuing calibration verification (CCV) standards 4.3.5. QC R ecovery Sam ples Prepare QC recovery samples to represent every control matrix included in the study. Prepare at least one QC recovery sample in the low-range, and one in the high-range, to approximately backet the expected range o f analyte in the samples. For sets containing more than 20 samples, prepare an q recovery sample for every group of 10 additional samples (for example if one sethas24samples.,then.3levelsofQCrecoverysampleswillbeprepared) .Prepare these adtfidonal QC recovery samples to bracket the range of analyte found in the samples, as appropriate. . * Note; An example of a sample extraction worksheet is attached as Attachment 1. ' 4.4. Extraction Note: A. Preparation/conditioning ofSPE columns is described in Section 4.41 B. Silanizition ofpear-shaped flasks is described in Section 4.4.2. " " ' C EvalnanWstandardization o fSPE columns (when a different supplier is used etc.) is described in Section 4.4,3. ` a. Weigh approximately 1.0 g ( 0.05 g) of sample in to 20 mL polyethylene scintillation vial (Note: designated samples need to be fortified "with appropriate aliquots offortification standards). b. Add 10 mL of ACN (note: break-up any clumps using a spatula or giass rod), shake on a wrist-action shaker for30 minutes. ' Centre Analytical Laboratories, laa Study# 023-003 Page 12 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 88 o f 103 Page 336 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 > Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-L10 - . Cafe N o-: OOM-Or-Otni revision 2 c. Let the vials sit for - 5 minutes, carefully decant, directly on to a ''0 t disposable syringe bane! connected to a glass acredisc (Gelman) git4r " d. Transfer the filtrate to a conditioned SPE column (see Section 4 4 n r 'l the elute in a 125-mL "silanized"(see Section 4.42) pear-shaped flask * ^ Mote: The 20. mL. S. PE, colu. m. .n fits well inside-tIoe mouth (sleeve) o f the pear-shaped flask. Make sure that the pear-shaped flask is well supported and will not topple. No vacuum is needed for this step e. Add 10 mL of ACN and then 20 ml. of 90:10 ACN : 2% ascorbic MeOH to the SPE column and collect the eluate in to the sszn a?!<1 111 flask, combining the eluates. . pear siuPed Add -3-4 drop of l-octanol to the pear shaped flask.and rotary evaporate t nearAdfr\yonness (a minute amount o f I-octanol will remain) at a reduc"'da Por5S5urc g. Reconstitute by adding 2.0 mL methanol (final volume=2 mL) and Vi/ todissolve. TransfertheextracttoHPLCvialsusingdisposablepipets ^ 4.4.1 SPE Column Preparation - Pack 20-mL SPE tabes with 2 g carbon. Condition columns with lo mL f saturated ascorbic Kid in MeOH followed by 10 mL ACN. Discard all washes. Do not allow the column to dry. . Note: The SPE columns may be packed and conditioned at any point Usethevacuummanifoldtoconditionthecolumns. 3 * ' Donotlet theSPEcolumnstorundryatanytime.. 4.4.2 SUanizution o fPear-ShapedFlasks ' Silanize the pear-shaped flasks, before use, by rinsing with a 30% dimethvi dichlorosilane in toluene solution followed by a toluene rinse and finally " methanol rinse. y 4.4.3 Standardization o fSPE Columns When using carbon from a different^ supalier, SPE columns should be standardized in the followingmanner, prior to analyzing samples' a. Using a mixed standard with a concentration between 100 and 500 nz/mL (each of all eight analytes) follow the elution schemes as outlined in 2 and 3. Collect all eluting fractions. . steps Ceacre A nalytical Laboratories, la c Study 1 023-003 Page 13 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page-89 of 103 Page 337 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 C a m M ehod No. : 00M-O23003, revision 2 b. Collect a post-elutiou fraction, after step 3, eluting with an additional ' 0 mL of 90:10 (ACN : 2% ascorbic acid in MeOH). ~ c. Adjust the final volume ofall the fiaen'ons to 10 mL with methanol. d. Analyze all the fractions by LC/MS/MS. . e. If the target fraction contains a minimum of 85% of the resoective analytes, it may be considered acceptable, ' f. If the "post-elution" fiaction contains significant amount of analytes the target elution volume may be increased. ' ' ' 4.5. Analysis by LC/MS/MS .' 4.5.1. LC/MS/MS System and Operating Conditions (Electrospray) Mass Spec: Interface: Micromass Quattro Ultima (Micromass) Elestrospray (Micromass) Harvard infusion pomp . Computer: Software: COMPAQ Professional Workstation AP2Q0 Windows NT, MassLynx 33 HPLC: Hewlett Packard (HP) Series 1100 HP QnatPump HP VacuumDegasser HP Autosampler ' . . HP Column Oven ' ' Note: A 4 x 10 mm hypercam drop-in guard cartridge (Keystone, part # 844017-400) is attached on-line after the purge valve and before the'sample injector port to trap any residue contaminants that may be in the mobile phase and/or HPLC system. HPLC Column: Genesis C, (Jones Chromatography), 2.1 mm x 50 mm, 4M C olum n Temp-' 35" C ' InjectionVoL: lOpLMobile Phase (A):2 mM Ammonium Acetate inType I water Mobile-Phase (B):Methanol Time 0 0.4 t.0 7.0 75 9.0 95 135 14.0 %A - <0 60 10 to 0 0 60 60 ' 60 %B Flow R ate rmTA rim i 40 0 5 0 0 40 0 5 0 0 90 0 5 0 0 ' 90 0 5 0 0 too too 40 40 ' 40' 0500 0.400 0.400 0.400 0500 CotreAnalytical Laboratories Inc. Study * 023-003 P age 14 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page-90 of 103 Page 338 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 . LRN-U2849 Centre Study No.: 023-016 SponsorProtocol No: FACT-TOX-liO It may be necessary to adjust the HPLC gradient in orde- to n ,, msmnnent performance. Columns with different dimensions fe > , ,pnnuzs 30 nan) and also columns from different manufacturers (Kevro v ! ^ X etc.) can be used provided equivalent cfaromatogranhy is obta'inei' 3613511 C;' Ions monitored: Analvte Mode PFOS PFOSA ' PFOSAA PFSEA EtFOSE-OH poaa" M556 M57 THPFOS negative negative negative negative negative negative negative negative negative Parent Ion 499 493 534 526 630 413 556 570 427 Product Ion 99 78 526 169 59 369 498 419 80 Approximate 6.0 5.7 6.7 6.7 5.1 5.4 5.6 5.1 Note that the retention times may vary slightly, on a dav-tn_j . depending on the batch ofmobile phase, etc. * y o-day basis, 4.5.2, Example TuneFile Parameters The following values are provided as an example. Actual values m ' from instrument to instrument. Also these values may be changed ^ to time in order to optimize for greatest sensitivity. S 110131 "rae Tire mass spectrometer is tuned using a solution of each analyte at - 0 5 pgAnL, prepared via dilution of the stock solution in methanol The i is infused (using a T et--tar) at 10 pL/min into a 0.2 miymJ ! t! l 'ltl0 r m obfle p h ase co nsisting o f 4054 m e th a n o l a n d 6054 2 mM ammonium a r T f Tne analytes are initially timed for the parent ion and then tuned f a product ion. The optimized parameters are saved as a "tune file" t v file is then used duringroutine analysis. ' ^ tunc Analvte PFOS PFOSA PFOSAA PFOSEA Et-FOSE-OH POAA M556 M570 THPFOS Dwell fsl - 0.2 02 02 02 02 02 02 0.2 02 Centre Analytical labram e*, Inc. Study 023-003 Collision Energy (V) 43 28 20 28 31 11 23 20 35 gpnefV) 76 34 37 49 30 25 50 60 34 Page IS Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 91 of 103 Page 339 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 C=tre M eio d No. ; OOfcM2j-;o3, wtaoa2 Soarct Csputery Hesapole I Aperture 1 Hexanole2 SourceBlock Temp. Desolvanon Temp. Set 2_5<SkV 0.5 V 0.2 V 0.3 V IOOC ' 40QC Anabzer LMResl HMResl IEnergy 1 Entrance Eat LMRts2 HMJtes2 IEsagy2 Multiplier ' . ' ' Set 13.0 V 13.0 V 0.7V ' -2 V IV 11.0 V 11.0 V 1.0 V 650 V Gas flows Cone Gas - Desolvanon Pressures Gas Cell Set -150 Lin~700L/hr Read h a r t ~ 3.0e-3 mbar N ote An alternative LC/MS/MS system may be used ones demonstrated . to beequivalent. ` ea 4.5,3. CalibndmProcedures a. Inject an afiquot (10 |iL) o f each calibration standard into the LC/MS/MS system. Inchtde standards corresponding to six or more conesatraiio levels (ranging t e n the LOQ to the highest concentration W ei to be included in theanalysis) in an analytical set. . b. Use a linear y = mx + b function for quantification. T in -,. Mim<ra n ] curves are geserated for each analyte by linear regression using l/x weighting ofpeak area versus calibration standard concentration usin* the Windows NI, MsssLynx 3.3 (or equivalent) software system. Anv extracted standards that fsll outside the 70 to 13054, based on the averaa" response (actor of the surrogate standard, must he excluded from* the calibration carve. C entre A nalytical Labotntoriea; tec. S b d y 9 023-003 Page IS Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 92 of 103 Page 340 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Catte MhodNo. : 0CM-3-CO3, revision i c. The correlation caezndat (R) for calibration curves generatedmust be (RJ2s0.96). If calibration results 01 outside these limits talce appropriate steps to adjust instrument operation, and reanalyze the relevant sets ofsamples. .. `` 4.5.4. SampleAnalysis . a. Each set of samples analyzed must mclude matrix blanks (witho surrogate standard), martrix zero blanks (with the surrogate standard) set ofextracted standards, and QC recovery samples. "3 b. Inject an aliquot (10 pL) of each standard/sampie/QC reeovety/control into the LC/MS/MS system. Begin a set ofsamples with an entire se* of calibrations. Include 'two continuing calibration verification fCCVY standards (one from the low-mid and one fiom the mid-high nme) aft . eve^ 5to 10 samples, at the most 61 c. Detennine die concentration o f each sampWQC recovery/control fiom the standard curve, based on the peak area of each analyte. The standard responses must bracket responses of the residue found in each sample s t Samples that fall outside the range of the standard curve must be appropriately andre-analyzed. " d. Evaluate the ongoing acceptability of instrumental analysis based on the CCVresults. Simples analyzed wiU be acceptable as long as hath CCV's fail within 30% of their true value. Calculate the CCV concentration found using the Imear calibration curve. If the CCV recovery is outside 70% to 130%, re-analyze samples analyzed afterthe last acceptable cW fc e. Monitor the response of the surrogate standard (SS) for every samnle extracted standard, blank; and QC recovery sample. The response is calculated using the "average response factor" junction using MassLynx 33 (or equivalent). Evaluate the acceptability of the sample extraction process based on the concentration of the SS found in all extracted standards, QC samples, blanks, and samples. The average response factor for all samples, blanks, extracted standards, and QC recovery samples must fill between 60% to 140% ( 40%). Those deviating by greater than 30% (outside 70% to 130%range) must be checked for possible errors and may need to be re-exltscred. Any responsesdeviating by greater than 40*/ (Ming outside the 60% to 140% range) will require re-extraction. If analysis is delayed, samples must be stored refiigerated at approximately 2*C to 6*C until analysis, and analyzed preferably within a week. Cemre Analytical Lcbonwics, Inc.Studyi 023-003 - Centre Analytical Laboratories, Inc . 3M Environmental Laboratory Page 93 of 103 Page 341 3M Medical Department Study. T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 4.6. Performance Criteria. . The following two criteria should be met before the inicial analysis cf samnles especially when using different instrumentation set-ups than those cit'd in" this method. First Criterion - Run a standard solution on LC/MS/MS corresponding to th estimated LOQ (the 10 ppb extracted standard) and obtain a signa'l to noise ratio of at least 9:1 for aD'analytes. I f this criterion cannot be met, optimize --d ch instrument operating parameter ` ge Second Criterion - Run a set of standards of six or more concentration levels ranging ftom the proposed LOQ op to the highest concentration eve! to be included in the analysis. Generate a calibration curve for each analyte and*obtain a linear regression with a correlationcoefficient of at least 0.98 for each analyte 4.7. Time Required for Analysis A set of 14 samples can iz taken through the extraction procedure in . approximately six hours by ene person. The LC/MS/MS analysis (P - M standards and 14 samples) will take approximately seven hours. ^" . 5- CALCULATIONS 5.1. Ana ly teFound . Calculate the amount o f analyte found (in ppb) using the s ta n d a rd curve generated by the Mass Lynx software program using Equation 1. Correction for n m ie weight (1 g) and final volume (2 mL) is not required since the samnles and standards arcextractedthesameway. ` Equation 1: (peak area-intercept) xDF Analyte found (ppb) = slope Where DF = dilution factor, if samples were diluted. Cearre Analytical Laboratories, lac. Study %023-003 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page IS Page 94 of 103 Page 342 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 . LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 C a s t Method N o. : OOM-O' l-OOl, z s r s io a 2 5.2. QC Recovery For samples fortined with known amounts of analytes prior to extracrion calculate the percent recover from Equation 2. ' Equation 2: %Recover = . analyte found (ppb) - analytefound inmatrixblank (ppb) amountanalyteadded (ppb) . 5.3. m ean Response Factor The mean response 6 ctor (MRF) is calculated by the Mass Lynx software program by averaging the mean response for all calibration standards (response/amount added) and dividing by the amount added. The actual ' concentration ftom each injection is then calculated by dividing the individual response by the MRF. The recovery for each injection is the percent of the j concentration to the amount added. This calculation is performed by the MassLynx 3j (or equivalent) software. 6. SAFETY ,, All samples must be treated as potential biohazards and appropriate Universal ' precautions'must be taken (fcilt/W standard operating procedures for the handling o f biofluids). These include, but are not Smiled to, the use o f double layers of latex or vinyl gloves, goggles, dust mask, and lab coat All disposable materials must be traded as biohazards and handled accordingly. : '' ' Acetonitrile, methanol, dimethyldichlorosilane and toluene are highly flammable. Open and use under fume hood only. All organic, waste and solvent waste must be segregated and disposed of accordingly. . Centre AnalyticalLabontoric, Inc. Sttaiy- 023-003 Page 19 [ ! i ! I j !i j I I ! ! j 1 t jt Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 95 of 103 Page 343 3M Medical Department Study. T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 ' Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 ATTACHMENT!. Css MebodNa.: 0OM-523-003, revision : / S X C entre Analytical Laboratories. Inc iriffniidirri-i *-- **,*"1 Fbor<iupJMa f*e(sn)2JW2sj o_r FSEFRATION o f samples f o r extraction and analysis FmocolH*: NA Coo*StudyHo * M*DrFtef MethodNo HA A la to FTOS.ffOSA. POAA.2-fOSE-QH.PfOSEA.FTOSAA. M3T0. m rtia e ^ L141U M upta" Sponsor Simple CcanStaph Sinta Wcfeb Fon. VoL Standard ID S> Dcscnp* U) m ID Mam fatano " 1 tMtms 309oust >-- * aio t u> j iw 1 UJ y j ""^uw j- " M 4 ~ n JW-- ' aio a ' 400 1 i ~ 1 aw 4 1 11 __________ !___________ 1 1 i t_________ . --------!-----------!-------------- 1 11 F n V.L Fw.ln.1 (pf/mt) . Wj Im H ? 1* vU ! * i------TO-- <U *W 1' iti l/J a i-- ss-- ik .Ui 1 IBS----- 2 H -- no-- .0 IQJJ . !-----i-----550-- tw 1 Juud ' l .. 1 1 1 .- _____ !____ - I _________ 1' i I1 1 1 1 1 1 11 i1 1 1 - H. i1 ~ 1----------------ii------------------- 1 i-- ; i -- i-------------- 1 1 ii -- ,-------------- 1' ` Jforjffedwide (bidsIs/Dais; Csncirr: / j.r SampkTemmrt<fm6tBEl---- Sanjrfaero*TjJ>cdooaboe*F_ Snpkjrerunedl* Ssreff F_____ "SunqpuStdlDs Sarnia taetd vidiAOiadSteri: CubonSPS clcan-upandM*np: Final folvmo adjustedto 2mLwvfcimcomI 2*oao*pbced ioreft(mori ----_ _/_ *tt|M U4 J _ minias /_ CUMAoote CirUI3JV40S AMfaAdd Mokwt l-OOMal j cw 1 TIkm J . CentreAnalytical Laboratories, Inc. StudyXG23*C03 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 20 Page 96 of 103 Page 344 3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 3048 Research Drive. Slate College. PA 16801. Phone; (814) 231-3032.Facsimile: METHOD DEVIATION Deviation Number: I Pase I of 2 DateofOccurrence: (1) 6/14/00, (2) 6/16/00, (3) 6/19/00, (4,5, &6). 6/22/00 m 6plm (8) 6/2S/00, (9) 6/29/00, (10) entire study ' Centre StudyNumber 023-016 Sponsor Protocol Number FACT-IOX-UO ***^jug ^xxcvimvmi newt** w w iu -u w -v t/j, ic v u io n .} 1. Section 4.4.a: Only033 g was weighed for Centre sample ID0003997 because that was all of thesample available: - Section 4.4j : Only0.70 g was weighed for Centre sample 0004031 and only 0.43 for 0004040 because that was all of the sample available. * 3. Section 4.4ji : Only031 gwas weighed for Centre sample ID0004032 because that was all ofthesample available. ' 4. Section 4.5,4 (e): Accepted recovery ofTHPFOS of 4356 for0003989 Spk A in Sec 062100AD. 5- Section 4.4j : Only0.70g was weighed for Centre sample ID 0004097 because that was all of the sample available. 6. Section 4.4.a: THPFOS was inadvertentlynot'spiked Into samples 0004097,0004098 . 0004099, and 0004100 in Set 062200A. , - 7, Section 4.4.a: Only057 g was weighed for Centre sample ID000417/ because that was ail of thesampleavailable. 5. Section 4.4j : Only0.66 g and 0.S8g was weighed for Centre sample ID's 0004186 and 0004194, respectively, because that was all of the sample available. 9- Section 4.4.a: Only0.30 g was weighed for Centre sample ID0004132 Spk A because that was all of the sample available. 10. Section 4.4.b.: Occasionally, some samples that contained haul fecal matter did not completelydisintegrateafter shaking. Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 97 f 103 Page 345 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Centre Analytical Laboratories, Inc. _304g R a c rd i Drive. Slate C alle. PA 16801. Phone: f31.fl231-3031 Fim im i],. m m , METHOD DEVIATION DeviationNumbcn 1 Page 2 o f2 DaeofOccurrence: (1) 6/14/00. (2) 6/16/00, (3) 6/19/00, (4.5. &6). 6/22/00 m 6D 7/ m (8) 6/23/00, (9) 6/29/00, (10) entire study ' Centre StudyNumber 023-016 Sponsor Protocal Number FACT-TOX-110 ACTIONSTAKEN i.e.. intendment issued. SOP revision, etc. 1 1-10. Methoddeviation issued. RecordedBy/Dlte; T tf'Soloh IMPACT ONTHESTUDY 1-j, 3,7-9 No negative impact 4. Nonegativeimpact because otherSpkwas acceptableand PFOS recovery was acce-table 6- No negative impact because THPFOS is not used forquantitative purposes andahTpFOs'' residues found weresimilar to the othersamples in theset 10. Nonegative impact becauseresidues found were consistent Principal Investigator Signature <$2? ix./*- Date Cesure QAU Review A/irw- Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 98 of 103 Page 346 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 3048 Research D rite,Slate CoUeje, PA ItiWL . Phone: (534)231-3032, Facsimile: (81-4)231-1253 'Tjz 6 d > Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 99 of 103 Page 347 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Centre Analytical Labor atopies, Inc. 3 0 Research Drive, Stale Colle-e, PA 14801. Phone: (814) 231-3032, Facsimile: <314)231-1253 . METHOD DEVIATION DeviationNumber 3 Date ofOccurrence: (I) 6/29-30/00 Centre Study Number: 023-016 Sponsor Protocol Number FACT-TOX-UO DESCRIPTIONOF DEVIATION Deviations to method titled"Determination ofFluorochemtcal Residues in Monkey/Rat Feces by LOMS/MS (Revision2)" (Centre Method 00M-O23-CO3. revision 2) 1, Section4.3.5: Didnot fortify any of the control samples fromthelast interval (GD11) of the study. ACTIONS TAKEN i.e-- amendment issued. SO P revision, etc. 1. Method deviationissued. Recorded By/Date: fy jp c L IMPACT ONTHE STUDY 1. Nonegative impact. Principal Investigator Signature Date Centre QAU Review v ti. February 12. 1998/2 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 100 of 103 Page 348 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Centre A nalytical Laboratories, Inc. Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 101 of 103 Page 349 3M Medical Department Study. T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Centre -Analytical Laboratories, Inc. 3043 Research Drive, State Caliese, PA 16301. Phone: (314)231-3032,Facsimile: (314)231-1253 m eth o d m o d ific a tio n fo r m . Modification No.: 2 Study Director: Marvin T. Case Page 1of 2 Centre StudyNnmher 023-016 SponsorProtocol Number: FACT-TOX-110 MethodTitle- DeterminationofFiuorochemical Residues inMonkey/Rat Feces by LC/MS/MS ________ (Revision2) Centre Method Number QQM-023-003. revision 2________ _______ MODIFICATION: 1. Section4.5.4 Sample Analysis (Iteme.) Should read as follows: Monitorthe response of the surrogate standard(SS) for everysample, extracted standard, blank, and QCrecoverysample. The concentration is calculated using the "average response factor" function fromMassLynx 3.3 software (or equivalent). Evaluate the acceptability of the sample extractionprocess based bn the concentrationofthe SS found in ail extracted standards, QC samples, blanks, and samples. The percent recovery for all samples, blanks, extracted standards. . and QCrecovery samples must fall between 60%and 140%. Those deviating by greater than 30%mnstbe checkedforpossible'errors and mayneed rtf-extracted. Any recoveries deviating bygreater than40%(Ming outside the60% to 140%range) will require re-extraction. 2. Section 3.1 AnalyteFound Remove secondsentence. 3. Section 5.1 Equations ModifyEquation 1to: . . Analyte found (ng/mL) =>fPeak area - intercept) slope Change Equation2 to: Analyte found(ppb)=*fanaivte found,fng/mL) x final voi. (mL) x DF) sample wt. (g) AddEquation 3: . . Recovery(%) = [analytefound (ngfmlQx final vol.(mL)xDF] analyteadded (ng) 4. Section S3 MeanResponse Factor Modifyfirst sentenceto read: . The mean response factor (MRF) is calculatedby the MassLynxsoftware programby averaging the response forall calibtation standards and dividing by the amount added. Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 1G2 of 103 Page 350 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Centre Analytical Laboratories, Inc. 3048 Research D rire, State Collese, PA 16801. Phone: (814)211-8032, Facsimile: (814)231.1233 m ethod m odification form Page 2 of 2 Modification No.: 2 StudyDirector MarvinT. Cue Centre StudyNumber 023-016 SponsorProtocol Number FACT-TOX-1I0 Method Title? Determination ofFluotoehemical Residues inMonlcey/RatFeces by LC/MS/MS ___ ______ (Revision 21 Centre MethodNumber OQM-023-003, revision 2_______ mSHFICATTON- 1. To clarifySS acceptable criteria. 2. Remove incorrect statement becausecalculations did include sample weight and final volume. 3. Modify calculations to correspond with those used instudy. 4. Clarify calculation ofmeanresponsefactor_______________' EFFECT ONSTUDY: 1*4. No negative impact. Originator o: Principal Investigatory Sponsor Management: \ ty l'/o n Dace r ( "( Date J-2/ku. Date February 12,1998/3 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 103 of 103 Page 351 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Analytical Laboratories, Inc:. 3048 Research Drive, State College, PA 16801. Phone: (814) 231-8032, Facsimile: (814)231-1253 21 Amended pages + amendment form ANALYTICAL PHASE REPORT AMENDMENT Amendment Number: 1 Effective Date: 9/20/00 Centre Study Number 023-016 Sponsor Protocol Number FACT-TOX-110 Report Title Oral (Gavage) Pharmacokinetic Study of PFOS in Rats Amended Section Section 1.0 Summary: 59386.8 ppb was changed to 59.4 pg/g in the third paragraph (See attached pagell). Section 7.6 Quantitation and Example Calculation: Equation 5: Residue Found (pg/g) = Residue Found (ng/g) * (1 pg /1000 ng) Equation 6: Corrected Residue Found (pg/g) = Residue Found (pg/g) * 0.869 These equations were only applied to the tables in the report. Section 8.0 Results: 59386.8 was changed to 59.4 pg/g in the second paragraph (See attached page 18). Section 11.0: Tables I - XXI (See attached pages 20-38) Reason for Amendment The units reported in the tables and text were changed from ng/g to pg/g, the number of significant figures represented in the tables and text was changed to three, the zeros reported in Tables II and XII were changed to ND's, and any value less than 0.0100 pg/g (LOQ) was reported as < 0.0100. No negative impact Impact on the Study Principal Investigator Signature VV-'-T' Study Director Signature CAL QAU Review Date 9 (h t Date ^ November 1995/0 3M Environmental Laboratory Page 352 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 - . Centre Study No.: 023-016 Sponsor Protocol No.: FACT-TOX-110 I 1.0 SUMMARY ! The purpose of this study was to analyze residues of perfluorooctanesulfonate (PFOS) in rat feces as specified in 3M Protocol FACT-TOX-110. The analytical method used for this study was entitled, "Determination of Fluorochemical Residues in Monkey/Rat Feces by LC/MS/MS, revision 2" (Centre method number: 00M-O23-O03, revision 2). The limit of quantification for PFOS in rat feces was 10 ppb (0.0100 pg/g). Residues (corrected for purity) ranging from non-detected levels to 59.4 pg/g were found in the rat feces samples. Fortification recoveries ranged from 71-128% with an average of 98% and relative standard deviation of 15%. 2.0 OBJECTIVE The objective of this study was to determine levels of perfluorooctanesulfonate (PFOS) in specimens of feces of rats using the analytical method entitled "Determination of Huorochemical Residues in Monkey/Rat Feces by LC/MS/MS, revision 2." ' 3.0 INTRODUCTION This report details the results of the residues of PFOS detected in rat feces, using the analytical method entitled, "Determination of Fluorochemical Residues in Monkey/Rat Feces by LC/MS/MS (revision 2)." Complete details of the analytical methodology can be found in Appendix B. . The study was initiated on June 10, 2000, when the study director signed the protocol FACT-TOX-110 Amendment #4. The complete protocol and amendments can be found in Appendix A. The analytical start date was June 9,2000, and the analytical termination date was July 1,2000. 4.0 TEST SYSTEM The 250 rat feces samples analyzed in this study were received frozen on dry ice from 3M Environmental Laboratory St. Paul, MN on May 26,2000 and stored frozen upon receipt. The samples were then logged in on May 31,2000 by Centre personnel and transferred to different frozen storage locale. The control rat feces used for standards and blanks was purchased from Lampire Biological Laboratories, Inc., Pipersville, PA and received at Centre frozen on dry ice on 3M Environmental Laboratory Amended Page 11 of 103 Page 353 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 - :Centre Study No.: 023-016 Sponsor Protocol No.: FACT-TO X -110 8.0 RESULTS AND DISCUSSION Although, the majority of the control samples did not contain, significant interferences, a couple of samples did contain residues that were comparable to control group levels. A summary of residues found in all of the matrix blanks and matrix zero blanks is detailed in Table I. Residues (corrected for purity) ranging from non-detected levels to 59.4 pg/g were found in the rat feces samples. The residues found in all of the samples plus the averages and standard deviations for each group at each interval are detailed in Tables II-XXI. For this report the residues found in Tables I-XXI were corrected for the purity of the PFOS standard lot 217 based on the certificate of analysis finalized on September 7, 2000. .' Fortification recoveries ranged from 71-128% with an average of 98% and relative standard deviation of 15% (n = 34). A summary of all of the fortification recoveries can be found in Table XXIL Typical calibration curves and chromatograms representing standards, controls, fortifications, and samples are depicted in Figures 1-7. 9.0 CIRCUMSTANCES THAT MAY HAVE AFFECTED THF. DATA Electronic records are not fully compliant with 21 CFR 11, "Electronic Records: Electronic Signature." However, fully approved SOP's were in place and all chromatography instrumentation used in this study was fully validated (IQ, PQ, OQ), calibrated and operational. All original data were printed as hard copies and fully audited by quality assurance. Verified exact copies and the electronic data have been stored- in the archives at Centre Analytical Laboratories, Inc. Original raw data have been returned to the sponsor. . 10.0 RETENTION OF DATAAND SAMPLES When the final report is complete, all original paper data generated by Centre Analytical Laboratories, Inc. will be shipped to the sponsor. This does not include facility-specific raw data such as instrument logs, however exact copies of temperature logs will be submitted. Exact copies of all raw data, as well as a signed copy of the final analytical, report and all original facility-specific raw data, will be retained in the Centre Analytical Laboratories, Inc. archives for the period of time specified in 21 CFR Part 58. Retained samples of reference substances are archived by the sponsor. 3M Environmental Laboratory Amended Page 18 of 103 Page 354 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849- Centre Study No.: 023-016 Sponsor Protocol No.: FACT-TOX-110 Table I. Summary of PFOS residues in M atrix Blanks and M atrix Zero Blanks ND = Not Detected and NQ = Not Quantifiable Sponsor Centre Residue Corrected Set Date Found Residue Found ID ID Number Extracted fug/.?) (ue/?) na 0004277 Blank A 061200AR 6/12/00 ND ND na 0004277 Blank B 061200AR 6/12/00 ND ND na 0004277Zero Blank C 061200AR 6/12/00 ND ND na 0004277 Zero Blank D 061200AR 6/12/00 ND ND na 0004277 BlankA 061400A 6/14/00 NQ NQ na 0004277 BlankB 061400A 6/14/00. NQ NQ na 0004277 Zero Blank C 061400A 6/14/00 NQ NQ na 0004277 Zero Blank D 061400A 6/14/00 ND ND na 0004277 Blank A 063500AD 6/15/00 NQ NQ na 0004277 BlankB 061500AD 6/15/00 NQ NQ na 0004277 Zero Blank C 061500AD 6/15/00 <0.0100 < 0.00869 na 0004277 Zero Blank D i 061SOOAD 6/15/00 <0.0100 <0.00869 na 0004277 BlankE 061600A 6/16/00 <0.0100 <0.00869 na 0004277 BlankF : 061600A 6/16/00 NQ NQ na 0004277 Zero Blank G 061600A 6/16/00 NQ NQ na - 0004277 Zero Blank H 061600A 6/16/00 NQ NQ na 0004277 Blank A 1 061900AD 6/19/00 NQ NQ na 0004277 BlankB 061900AD 6/19/00 NQ NQ na 0004277 Zero Blank C 061900AD 6/19/00 NQ NQ na 0004277 Zero Blank D 061900AD 6/19/00 . nq NQ na 0003466 BlankA 062000A 6/20/00 NQ NQ na 0003466 BlankB 062000A 6/20/00 NQ NQ na 0003466 Zero Blank C 062000A 6/20/00 NQ NQ na 0003466 Zero Blank D 062000A 6/20/00 NQ NQ na 004277Blank A 062100A 6/21/00 NQ NQ na 0004277 BlankB 062100A 6/21/00 NQ NQ na 0004277 ZeroBlank C 062100A 6/21/00 NQ NQ na 0004277 Zero Blank D 062100A 6/21/00 NQ NQ na 0004277 BlankA 062200A 6/22/00 <0.0100 <0.00869 na 0004277 BlankB 062200A 6/22AX) ND ND na 0004277 Zero Blank C 062200A 6/22/00 ND ND na 0004277 Zero BlankD- 062200A 6/22/00 ND ND na 0004277 BlankE 062200B 6/22/00 ND ND na 0004277 BlankF 062200B 6/22/00 ND ND na 0004277 Zero Blank G 062200B 6/22/00 ND ND 3M Environmental Laboratory Amended Page 20 of 103 Page 355 3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849- Centre Study No.: 023-016 Sponsor Protocol No.: FACT-TOX-110 Table I (cont.) Summary of PFOS residues in M atrix Blanks and M atrix Zero Blanks ND = Not Detected and NQ = Not Quantifiable Sponsor Centre ID Set _ Residue Corrected Residue Date Found Found ID Number Extracted (u.2/2) (us/g) na 0004277 Zero Blank H 062200B 6/22/00 ND ND na 0004277 Blank A 062300A 6/23/00 ND ND na 0004277 Blank B 062300A 6/23/00 ND ND na 0004277 Zero Blank C 062300A 6/23/00 ND ND na 0004277 Zero Blank D Q62300A 6/23/00 ND ND na 0004277 Blank A 062300B 6/23/00 NQ NQ na 0004277 BlankB 062300B 6/23/00 NQ NQ na 0004277 Zero Blank C 062300B 6/23/00 ' NQ NQ na 0004277 Zero Blank D 062300B 6/23/00 NQ NQ na 0005738 Blank A 062600A 6/26/00 <0.0100 < 0.00869 na 0005738 BlankB 062600A 6/26/00 0D126 0.0109 na 0005738 Zero Blank C 062600A 6/26/00 ND . ND na 0005738 Zero BlankD 062600A 6/26/00 ND ND na 0005738 BlankE 062600B 6/26/00 ND ND na 0005738 BlankF 062600B 6/26/00 ND . ND ~ na 0005738 ZeroBlank G 062600B 6/26/00 ND ND na 0005738 Zero Blank H 062600B 6/26/00 ND ND na 0005738 Blank A 062700A 6/27/00 ND ND na 0005738 Blank B 062700A 6/27/00 ND ND na 0005738 Zero Blank C 062700A 6/27/00 ND ND na 0005738 Zero Blank D 062700A 6/27/00 ND ND na 0005738 BlankA 062800A 6/28/00 ND ND na .0005738 BlankB 062800A 6/28/00 ND ND na 0005738 Zero Blank C 062800A 6/28/00 ND . ND na 0005738 Zero BlankD 062800A 6/28/00 ND ND na 0005738 Blank A 062900A 6/29/00 0.0301 0.0262 na 0005738 Blank B 062900A 6/29/00 ND ND na 0005738 Zero Blank C 062900A 6/29/00 ND ND na 0005738 Zero Blank D 062900A 6/29/00 ND ND na 0005738 Blank A 063000A 6/30/00 - ND ' ND na 0005738 Blank B 063000A 6/30/00 ' ND ND na 0005738 Zero Blank C 063000A 6/30/00 ND ND na 0005738 Zero Blank D 063000A 6/30/00 ND . ND AVERAGE: < 0.0100 <0.00869 STANDARD. DEVIATION: NQ NQ Note: The average and standard deviation were calculated by using 0.0100 as the value for those samples reported as < 0.0100,0.00869 for those reported as < 0.00869, and zero for those reported as ND or NQ. Amended Page 21 of 103 Laboratory Page 356 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 - ' Centre Study No.: 023-016 > Sponsor Protocol No.: FACT-TOX-110 Table II. Summary o f PFOS residues in Gl-Control-GDO Sponsor ED 19501FP0,Gl-Control-GDO 19502F.F0,Gl-Control-GDO 19503F,F0,Gl-Control-GD0 19504FPO,Gl-Control-GDO 19505FP0,Gl-Control-GDO 195O6FJF0,Gl-Control-GDO 19507FJF0,Gl-Control-GDO 19508F.F0,Gl-Control-GDO 19509FP0,Gl-Control-GDO 195lOFpO,Gl-Control-GDO 19511FJF0,Gl-Control-GDO 19512FJF0,Gl-Control-GDO 19513F.F0,Gl-Control-GDO 19514F.F0,Gl-Control-GDO 19515FP0,Gl-Control-GDO 19516FJF0,Gl-Control-GD0 Centre ED i Set Corrected Date Residue Found Residue Found Number Extracted 0003977 061200AR 6/12/00 0003978 061200AR 6/ 12/00 <U2/g) ND ND (U.2 /s) ND ND 0003979 061200AR 6/ 12/00 0003980 061200AR 6/ 12/00 < 0.0100 ND <0.00869 ND 0003981 061200AR 0003982 061200AR 0003983 061200AR 6/12/00 6/12/00 6/12/00 ND 0.0301 ND ND 0.0262 ND 0003984 061200AR 6/12/00 < 0 .0 10 0 < 0.00869 0003985 061200AR 6/12/00 0.0400 0.0348 0003986 061200AR 6/12/00 . ND ND 0003987 061200AR 6/12/00 ND ND 0003988 061200AR 6/12/00 ND ND 0003989 061200AR 6/12/00 ND ND 0003990 061200AR 6/ 12/00 0003991 061200AR 6/12/00 ND ND ND ND 0003992 061200AR 6/12/00 0.0609 0.0529 AVERAGE: < 0.0100 < 0.00869 STANDARD DEVIATION: NQ NQ ND = Not Detected and NQ = Not Quantifiable Note: The average and standard deviation were calculated by using 0.0100 as the value for those samples reported as < 0.0100,0.00869 for those reported as < 0.00869, and zero for those reported as ND or NQ- . 3M Environmental Laboratory Amended Page 22 of 103 Page 357 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 . LRN-U2849- Centre Study No.: 023-016 Sponsor Protocol No.: FACT-TOX-110 Table III. Summary of PFOS residues in G2-0.1MKD on GDO Sponsor Centre ID Set Corrected Date Residue Found Residue Found ID 19517F,F0,G2-0.1MKD-GD0 0003993 19518F,F0,G2-0.1MKD-GD0 0003994 19519FF0,G2-0.1MKD-GD0 0003995 19520F,FO,G2-0.1MKD-GD0 0003996 Number 061400AD 061400A 061400A 061400AD Extracted 6/14/00 6/14/00 6/14/00 6/14/00 (ne/s) 0.701 0.456 0.390 0.676 (uafs) 0.609 0.396 0.339 0.587 1952lF,F0,G2-0.1MKD-GD0 0003997 061400A 6/14/00 0.495 0.430 19522F,F0,G2-0.1MKD-GD0 0003998 061400A 6/14/00 0.381 0.331 19523F,F0,G2-0.1MKD-GD0 0003999 061400A 6/14/00 . 0.458 0.398 19524F,F0,G2-0.1MKD-GD0 0004000 061400AD 6/14/00 0.833 ' 0.724 19525F,F0,G2-0.1MKD-GD0 0004001 061400A 6/14/00 0.473 0.411 19526FP0.G2-0.1MKD-GD0 0004002 061400A 6/14/00 0.446 0.388 19527F,F0,G2-0.1MKD-GD0 0004003 061400A 6/14/00 0.497 19528FJF0.G2-0.1MKD-GD0 0004004 061400AD 6/14/00 0.701 0.432 0.609 19529FP0.G2-0.1MKD-GD0 0004005 061400AD 6/14/00 . 0.797 0.693 19530F.F0.G2-0.1MKD-GD0 0004006 061400A 6/14/00 0.394 ,0.342 1953lF,F0,G2-0.1MKD-GD0 0004007 061400AD 6/14/00 0.735 0.638 19532F.F0.G2-0.1MKD-GD0 0004008 061400AD 6/14/00 0.748 0.650 AVERAGE: 0.574 0.499 STANDARD DEVIATION: 0.160 0.139 3M Environmental Labmatury Amended Page 23 of 103 Page 358 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849- | Centre Study No.: 023-016 : Sponsor Protocol No.: FACT-TOX-110 Table IV. Summary of PFOS residues in G3-0.4MKD on GDO 1 i Sponsor Centre ID Set Corrected Date Residue Found Residue Found: ID 1 Number Extracted fpz/s) (112/2 ) 19533Fp0,G3-O.4MKD-GD0 0004009 061500AD 6/15/00 2.60 2.26 19534F,F0,G3-0.4MKD-GD0 0004010 061500AD 6/15/00 3.75 3.26 19535F,F0,G3-0.4MKD-GD0 0004011 061500AD 6/15/00 2.80 2.43 19536FVF0,G3-0.4MKD-GD0 0004012 061500AD 6/15/00 3.17 2.76 19537FJF0.G3-0.4MKD-GD0 0004013 061500AD 6/15/00 2.78 2.41 19538FP0.G3-0.4MKD-GD0 0004014 061500AD 6/15/00 3.08 2 .6 8 19539F,FO,G3-O.4MKD-GD0 0004015 061500AD 6/15/00 3.68 3.20 1954OFJF0.G3-O.4MKD-GD0 0004016 061500AD 6/15/00 2.04 1.78 19541F,F0,G3-0.4MKD-GD0 0004017 061500AD 6/15/00 ' 2 .1 0 1.83 19542F.F0.G3-0.4MKD-GD0 0004018 061500AD 6/15/00 3.29 2.86 19543FJF0.G3-0.4MKD-GD0 0004019 06150AD 6/15/00 2 .1 1 1.84 19544FJFO.G3-0.4MKD-GDO 0004020 061500AD 6/15/00 3.43 2.98 19545F,F0,G3-0.4MKD-GD0 0004021 061600AD 6/16/00 2.76 2.39 19546F,F0,G3-0;4MKD-GD0 0004022 061500AD 6/15/00 2.33 2.03 19547F,F0,G3-0.4MKD-GD0 0004023 061500AD 6/15/00 2 .2 2 1.93 19548Fp0.G3-O.4MKD-GDO 0004024 061500AD 6/15/00 2.33 2.03 AVERAGE: 2.78 2.42 STANDARD DEVIATION: 0.570 0.496 3M Environmental Laboratory Amended Page 24 of 103 Page 359 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849. . : Centre Study No.: 023-016 , ' Sponsor Protocol No.: FACT-TOX-110 Table V. Summary of PFOS residues in G4-1.6MKD on GDO t Sponsor , Centre ID Set Corrected Date Residue Found Residue Found ID Number Extracted 19549FP0.G4-1.6MKD-GD0 0004025 061600AD 6/16/00 fu.a/a) 14.1 (pa/a) 12.3 19550F,F0,G4-1.6MKD-GD0 0004026 061600AD 6/16/00 16.9 14.7 19551FJF0.G4-1.6MKD-GD0 0004027 061600AD 6/16/00 10.4 9.03 19552FJ0.G4-1.6MKD-GD0 0004028 061600AD 6/16/00 13.1 11.4 19553FJF0.G4-1.6MKD-GD0 0004029 061600AD 6/16/00 13.7 11.9 19554FJO,G4-1.6MKD-GD0 0004030 061600AD 6/16/00 15.5 13.5 19555FJF0.G4-1.6MKD-GD0 0004031 061600AD 6/16/00 10.3 8.98 19556FJ0.G4-1.6MKD-GD0 0004032 061600AD 6/16/00. 10.3 8.91 19557F,F0,G4-1.6MKD-GD0 0004033 061600AD 6/16/00 8.96 7.79 19558FJF0,G4-1.6MKD-GD0 0004034 061600AD 6/16/00 6.85 5.95 19559FJ0.G4-1.6MKD-GD0 0004035 061600AD 6/16/00 7.68 6.67 19560FJ0,G4-1.6MKD-GD0 0004036 061600AD 6/16/00 .16.5 14.4 19561F,F0,G4-1.6MKD-GD0 0004037 061600AD 6/16/00 1 2 .1 10.5 19562FJ0.G4-1.6MKD-GD0 0004038 061600AD 6/16/00 .9.59 8.33 19563F.F0.G4-1.6MKD-GD0 0004039 061600AD 6/16/00 7.47 ' 6.49 19564FJ0.G4-1.6MKD-GD0 0004040 061600AD 6/16/00 17.0 14.7 AVERAGE: 11.9 10-3 STANDARD DEVIATION: 3.45 3.00 3M Environmental Laboratory Amended Page 25 of 103 Page 360 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 . Centre Study No.: 023-016 Sponsor Protocol No.: FAcjrT-TOX-l 10 Table VI. Summary of PFOS residues in G5-3.2MKD on GDO Sponsor Centre ID Set Corrected Date Residue Found Residue Found ID Number Extracted . (ua/?) : (ua/a) 19565FP0.G5-3.2MKD-GD0 0004041 061900AD 6/19/00 23.5 20.4 19566F.F0.G5-3.2MKD-GD0 0004042 061900AD 6/19/00 29.5 25.7 19567FJF0.G5-3.2MKD-GD0 0004043 061900AD 6/19/00 30.3 26.3 19568F,F0,G5-3.2MKD-GD0 0004044 061900AD 6/19/00 24.2 2 1 .0 19569FJF0.G5-3.2MKD-GD0 0004045 061900AD 6/19/00 27.1 23.5 19570F.F0.G5-3.2MKD-GD0 0004046 061900AD 6/19/00 19.7 17.1 19571FJ0.G5-3.2MKD-GD0 0004047 061900AD 6/19/00 29.9 26.0 19572F,FO,G5'-3.2MKD-GDO 0004048 061900AD 6/19/00 25.4 2 2 .1 19573F,F0,G5-3.2MKD-GD0 0004049 061900AD 6/19/00 23.0 2 0 .0 19574F,F0,G5-3.2MKD-GD0 0004050 061900AD 6/19/00 29.3 25.5 19575F,F0,G5-3.2MKD-GD0 0004051 061900AD 6/19/00 23.0 2 0 .0 19576F,FO,G5-3.2MKD-GD0 0004052 061900AD 6/19/00 36.3 31.6 19577F,F0,G5-3.2MKD-GD0 0004053 061900AD 6/19/00 34.2 29.8 19578FJ0,G5-3.2MKD-GD0 0004054 061900AD 6/19/00 . ' 34.8 30.2 19579FJF0.G5-3.2MKD-GD0 Q004055 061900AD 6/19/00 25.4 2 2 .0 19580FJ0.G5-3.2MKD-GD0 0004056 061900AD 6/19/00 24.9 2 1.6 AVERAGE: 27 23.9 STANDARD DEVIATION: 4.78 4.15 3M Environmental Laboratory Amended Page 26 of 103 Page 361 3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 ............LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No.: FACT-TOX-110 Table VII. Summary o f PFOS residues in G l-Control on GD7 Sponsor Centre ID Set Corrected Date Residue Found Residue Found ' ID 19501F,F0,G1-Control-GD7 19502FJ0,Gl-ControI-GD7 19504FJ0.Gl-Control-GD7 0004057 0004058 0004059 Number 062000A 062000A 062000A Extracted 6/20/00 6/20/00 6 /2 0 / 0 0 (ua/e) ND NQ NQ (usJs) ND NQ NQ 19505F,F0,Gl-Control-GD7 0004060 062000A 6/20/00 ND ND 19506FJF0.G1-Control-GD7 0004061 062000A 6/20/00 0.0178 0.0155 19507F,F0,Gl-Control-GD7 0004062 062000A 6/20/00 ND ND 19508FJ0,Gl-Control-GD7 0004063 062000A 6/20/00 <0.0100 < 0.00869 19509F,F0,Gl-Control-GD7 0004064 062000A 6/20/00 ND ND 19510F,F0,Gl-Control-GD7 0004065 062000A 6/20/00 ND ND 19511FJF0.G1-Control-GD7 0004066 062000AR 6/20/00 ND ND 19512F,FO,G1-Control-GD7 0004067 062000AR 6/20/00 ND ND 19513FFO,G1-Control-GD7 0004068 062000A 6/20/00 ND ND 19514FJFO,G1-Control-GD7 0004069 062000A 6/20/00 ND ND 19515FJ0,Gl-Control-GD7 0004070 062000AR 6/20/00 ND ND 19516F,FO,G1-Control-GD7- 000471 062000AR 6/20/00 NQ NO AVERAGE: <0.0100 <0.00869 STANDARD DEVIATION: NQ NQ ND = Not Detected and NQ = Not Quantifiable Note: The average and standard deviation were calculated by using 0.0100 as the value for those samples reported as < 0.0100,0.00869 for those reported as < 0.00869, and zero for those reported as ND or NQ. 3M'Environmental Laboratory Amended Page 27 of 103 Page 362 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 ............. LRN-U2849- . ; Centre Study No.: 023-016 . Sponsor Protocol No.: FACT-TOX-110 Table VIII. Summary of PFOS residues in G2-0.1MKD on GD7 Sponsor Centre ID Set Corrected Date Residue Found Residue Found ID Number Extracted O12 /2 ) (PSt/fO 19517F,F0,G2-0.1MKD-GD7 0004072 062100A 6/21/00 0.409 0.356 19518F,F0,G2-0.1MKD-GD7 0004073 062100AD 6/21/00 0.561 0.488 19519FvF0,G2-0.1MKD-GD7 0004074 062100A 6/21/00 0.345 0.299 19521FJ0.G2-0.1MKD-GD7 0004075 062100A 6/21/00 0.465 0.404 19522FJF0.G2-0.1MKD-GD7 0004076 062100AD 6/21/00 0.778 0.676 19523FJF0,G2-0.1MKD-GD7 0004077 062100AD 6/21/00 0.625 0.543 19524FJF0,G2-0.1MKD-GD7 0004078 062100AD 6/21/00 0.576 0.500 19525F,F0,G2-0.1MKD-GD7 0004079 062100AD 6/21/00 ' 0.595 0.517 19526F,F0,G2-0.1MKD-GD7 0004080 062100A 6/21/00 0.469 0.408 19527FF0.G2-0.1MKD-GD7 0004081 062100AD 6/21/00 0.742 0.645 19528F,F0,G2-0.1MKD-GD7 0004082 062100AD 6/21/00 0.532 0.462 19529FJF0.G2-0.1MKD-GD7 0004083 062100AD 6/21/00 0.744 0.647 19530FFO,G2-0.1MKD-GD7 0004084 062100A 6/21/00 0.424 0.369 1953lFvF0,G2-0.1MKD-GD7 0004085 062100AD 6/21/00 0.624 0.542 19532F.F0.G2-0.1MKD-GD7 0004086 062100AD 6/21/00 0.574 . 0.499 AVERAGE: 0S64 0.490 . STANDARD DEVIATION: 0.128 0.111 3M Environmental Laboratory Amended Page 28 of 103 Page 363 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849... ! Centre Study No.: 023-016 Sponsor Protocol No.: FACT-TOX-110 Table IX. Summary o f PFOS residues in G3-0.4MKD on GD7 i Sponsor Centre ID Set Corrected Date Residue Found Residue Found ID Number Extracted (nzfg) (us/g) 19533FJD.G3-0.4MKD-GD7 0004087 062200AD 6/22/00 2.55 2.21 19534FJF0.G3-0.4MKD-GD7 0004088 062200AD 6/22/00 2.69 2.34 19535FJ0.G3-0.4MKD-GD7 0004089 062200AD 6/22/00 1.82 1.59 19536F,F0,G3-0.4MKD-GD7 0004090 062200AD 6/22/00 3.26 2.83 19537FJ0.G3-0.4MKD-GD7 0004091 062200AD 6/22/00 2.61 2.27 19538FJFO.G3-0.4MKD-GD7 0004092 062200AD mum 2.93 2.55 19539FJF0.G3-0.4MKD-GD7 0004093 062200AD mum 2.09 ' 1.82 19540FJF0.G3-0.4MKD-GD7 0004094 62200AD mum 2.27 1.97 19542FJ0.G3-0.4MKD-GDT 0004095 062200AD mum 3.00 2.61 19543FJ0.G3-0.4MKD-GD7 0004096 062200AD mum. 2.80 . 2.43 19544FF0.G3-0.4MKD-GD7 0004097 062200AD mum 2.28 1.98 19546F0.G3-0.4MKD-GD7 0004098 062200AD mum 1.52 1.32 19547FF0.G3-0.4MKD-GD7 0004099 062200AD 6/22/00 2.06 1.79 19548FJF0.G3-0.4MKD-GD7 0004100 062200AD mum 2.85 2.47 - '` AVERAGE: 2.48 ' 2.16 STANDARD DEVIATION: 0.492 0.428 3M Environmental Laboralory Amended Page 29 of 103 Page 364 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 : Centre Study No.: 023-016 Sponsor Protocol No.: FACT-TOX-110 Table X. Summary of PFOS residues in G4-1.6MKD on GD7 Sponsor Centre ID Set Corrected iDate Residue Found Residue Found ID 19550F,FO,G4-1.6MKD-GD7 0004101 19553FJ0.G4-1.6MKD-GD7 0004102 19554FF0.G4-1.6MKD-GD7 0004103 Number 062200BD 062200BD 062200BD Extracted 6/22/00 6/22/00 6/22/00 (P.e/g) 15.0 12.3 11.8 (U2/K) 13.0 10.7 10.2 19555F,F0,G4-1.6MKD-GD7 0004104 062200BD 6/22/00 9.61 8.35 19556FF0.G4-1.6MKD-GD7 0004105 062200BD 6/22/00 10.2 8.88 19558F.F0,G4-1.6MKD-GD7 0004106 062200BD 6/22/00 5.89 5.12 19559F,F0J4-1.6MKD-GD7 0004107 062200BD 6/22/00 6.89 5.99 19560FF0.G4-1.6MKD-GD7 0004108 062200BD 6/22/00 7.02 6.10 19561FF0.G4-1.6MKD-GD7 0004109 062200BD 6/22/00 8.88 7.71 I9562F.F0.G4-I.6MKD-GD7 0004110 062200BD ' 6/22/00 10.8 9.42 19563FF0.G4-1.6MKD-GD7 0004111 062200BD 6/22/00 14.9 13.0 19564FJ0.G4-1.6MKD-GD7 0004112 062200BD 6/22/00 13.7 11.9 AVERAGE: 10.6 9.19 STANDARD DEVIATION: 3.08 2.67 3M Environmental Laboratory Amended Page 30 of 103 Page 365 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN4J2849 ~ Centre Study No.: 023-016 Sponsor Protocol No.: FACT-TOX-110 Table XI. Summary of PFOS residues in G5-3.2MKD on GD7 Sponsor Centre ID Set Corrected Date Residue Found Residue Found ID Number Extracted 19565F.F0.G5-3.2MKD-GD7 0004113 062300AD 6/23/00 19566FJF0.G5-3.2MKD-GD7 0004114 062300AD 603100 (ue/s) 51.7 26.5 (us/s) 44.9 23.0 19567FP0.G5-3.2MKD-GD7 0004115 062300AD 6/23/00 35.9 31.2 19568F.F0.G5-3.2MKD-GD7 0004116 062300AD 6/23/00 23.0 20.0 19569FJF0.G5-3.2MKD-GD7 0004117 062300AD 6/23/00 42.9 37.3 19570FP0.G5-3.2MKD-GD7 0004118 062300AD 6/23/00 37.4 32.5 19571FP0.G5-3.2MKD-GD7 0004119 062300AD 603/00 43.0 37.4 19572FF0,G5-3.2MKD-GD7 0004120 062300AD 6/23/00 38.8 33.7 ' 19573FJ0.G5-3.2MKD-GD7 0004121 062300AD 6/23/00 26.7 23.2 19574F.F0,G5-3.2MKD-GD7 0004122 062300AD 6/23/00 36.3 31.6 19575FF0.G5-3.2MKD-GD7 0004123 062300AD 6/23/00 35.5 30.8 19577F,F0,G5-3.2MKD-GD7 0004124 062300AD 6/23/00 68.3 59.4 19579FP0.G5-3.2MKD-GD7 0004125 062300AD 6/23/00 35.9 31.2 19580FF0.G5-3.2MKD-GD7 0004126 062300AD 6/23/00 30.1 26.1 AVERAGE: 38.0 33.0 STANDARD DEVIATION: 11.5 10.0 3M Environmental Laboratory Amended Page 31 of 103 Page 366 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849- . , Centre Study No.: 023-016 , SponsorProtocol No.: FACT-TOX-110 Table XII. Summary o f PFOS residues in Gl-Control on GD15 Sponsor ID Centre IDi Set Number 19501FJF0,Gl-Control-GD15 0004127 062300B Corrected Date Residue Found Residue Found Extracted ... foste) (U/a) 6/23/00 ND ND 19502F,F0,G1-Control-GD15 0004128 062300B 6/23/00 ND ND 19504FP0,Gl-Control-GD 15 0004129 062300B 6/23/00 ND ND 19505FJ0,G1-Control-GD15 0004130 062300B 6/23/00 `ND ND 19506F,F0,Gl-ControI-GD15 0004131 ' 062300B 6/23/00 0.0445 0.0387 19507F,F0,G3-Control-GD15 0004132 062300B 6/23/00 ND ND I9508F,F0,Gl-Control-GD15 0004133. 062300B 6/23/0 ND ND 19509F,F0,Gl-ControMjD15 0004134 062300B 6/23/00 0.0242 0.0210 1951OFJO,G1-Control-GD15 0004135 0623OOB 6/23/00 <0.0100 <0.00869 19511FJO,GI-Control-GD15 0004136 062300B 6/23/00 ND ND 19512FJ0,Gl-Control-GD 15 0004137 062300B 6/23/00 0.0579 0.0503 19513F.F0,Gl-Control-GD15 0004138 062300B 6/23/00 ND ND 19514FJFO,Gl-Control-GD15 0004139 062300B 6/23/00 ND ND 19515F,FO,G1-Control-GD15' 0004140 062300B 6/23/00 ND ND 19516F,FO,G1-Control-GD15 0004141 062300B 6/23/00 0.0216 0.0188 AVERAGE: 0.0105 0.00917 STANDARD DEVIATION: 6.0185 0.0161 ND = Not Detected and NQ = Not Quantifiable . Note: The average and standard deviation were calculated by using 0.0100 as the value for those samples reported as < 0.0100,0.00869 for those reported as < 0.00869, and zero for those reported as ND or NQ. 3M Environmental Laboratory Amended Page 32 of 103 Page 367 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 ! Centre Study No.: 023-016 Sponsor Protocol No.: FACT-TOX-110 Table XIII. Summary of PFOS residues in G2-0.1MKD on GD15 Sponsor Centre JD Set Corrected Date Residue Found Residue Found ID Number Extracted 19517F,F0,G2-0.1MKD-GD15 0004142 062600AD 6/26/00 0s/f0 0.758 (U2fg) 0.659 19518F,F0,G2-0.1MKD-GD15 0004143 062600AD 6/26/00 0.672 0.584 19519F.F0.G2-O.1MKD-GD15 0004144 062600AD 6/26/00 0.782 0.680 19521F,F0,G2-0.1MKD-GD15 0004145 062600AD 6/26/00 0.632 0.549 19522F,F0,G2-0.1MKD-GD15 0004146 062600AD 6/26/00 0.817 0.710 19523FJ0,G2-0.1MKD-GD15 0004147 062600AD 6/26/00 0.642 0.558 19524FvF0,G2-0.1MKD-GD15 0004148 062600AD 6/26/00 ' 0.834 0.725 19525F.F0.G2-0.1MKD-GD15 0004149 062600AD 6/26/00 1.08 0.939 19526FJF0.G2-0.1MKD-GD15 0004150 062600AD 6/26/00 0.778 0.676 19527FJ0.G2-0.1MKD-GD15 0004151 062600AD 6/26/00 0.799 0.694 19528F,F0,G2-0.1MKD-GD15 0004152 062600AD 6/26/00 0.845 0.734 19529FF0.G2-0.1MKD-GD15 0004153 062600AD 6/26/00 0.724 0.629 19530FJ0.G2-O.1MKD-GD15 0004154 062600AD 6/26/00 0.724 0.630 1953lF,F0,G2-0.1MKD-GD15 0004155 062600AD 6/26/00 0.777 0.675 19532F.F0.G2-0.1MKD-GD15 0004156 062600AD 6/26/00 0.568 0.494 AVERAGE: . 0.762 0.662 STANDARD DEVIATION: : 0.119 0.104 3M Environmental Laboratory Amended Page 33 of 103 Page 368 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 ...... LRN-.U2849 . Centre Study No.: 023-01(5 Sponsor Protocol No.: FACT-TOX-llp Table XIV. Summary of PFOS residues in G3-0.4MKD on GD15 = Sponsor Centre ID Set Corrected Date Residue Found Residue Fpund ID Number Extracted . (Fft/fO (ue/g) 19533FF0.G3-0.4MKD-GD15 0004157 062600BD 6/26/00 2.76 2.40 19534F.F0.G3-0.4MKD-GD15 0004158 062600BD 6/26/00 4.82 4.19 19535F,F0,G3-O.4MKD-GD15 0004159 062600BD 6/26/00 3.63 3.15 19536FF0.G3-0.4MKD-GD15 0004160 062600BD 6/26/00 3.19 2.77 19537F,F0,G3-0.4MKD-GD15 0004161 062600BD 6/26/00 3.40 2.95 19538FF0.G3-0.4MKD-GD15 0004162 062600BD 6/26/00 3.14 2.73 19539F,F0,G3-0.4MKD-GD15 0004163 062600BD 6/26/00 3.09 2.69 19540FhF0,G3-0.4MKD-GD5 0004164 062600BD 6/26/00 4.15 3.61 19542F,F0,G3-0.4MKD-GD15 0004165 062600BD 6/26/00 1.88 1.64 19543F,F0,G3-0.4MKD-GD15 0004166 062600BD 6/26/00 3.42 2.97 19544F,F0,G3-0.4MKD-GD15 0004167 062600BD 6/26/00 3.37 2.93 19546F.F0.G3-0.4MKD-GD15 0004168 062600BD 6/26/00 2.63 2.28 19547FJ0.G3-0.4MKD-GD15 004169 062600BD 6/26/00 3.86 3.36 19548F,F0,G3-0.4MKD-GD15 0004170 062600BD 6/26/00 3.91 3.40 - - AVERAGE: 3.38 2.93 STANDARD DEVIATION: 0.713 0.620 3M Environmental Laboratory Amended Page 34 of 103 Page 369 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 .................. LRNi U2849--- i Centre Study No.: 023-bl6 , Sponsor Protocol No.: FACT-TOX-i 10 Table XV. Summary o f PFOS residues in G4-1.6MKD on GD15 . Sponsor , Centre ID Set Corrected Date Residue Found Residue Found ID Number Extracted (u.e/g) (ue/g) 19550FF0.G4-1.6MKD-GD15 0004171 062700AD 6/27/00 13.9 12.1 19553F,F0,G4-1.6MKD-GD15 0004172 062700AD 6/27/00 14.8 12.9 19554FF0.G4-1.6MKD-GD15 0004173 062700AD 6/27/00 14.2 12.3 19555F,F0,G4-1.6MKD-GD15 0004174 062700AD 6/27/00 16.3 14.1 19556F,F0,G4-1.6MKD-GD15 0004175 062700AD 6/27/00 9.07 7.88 19558FF0.G4-1.6MKD-GD15 0004176 062700AD 6/27/00 7.17 6.23 19559FF0.G4-1.6MKD-GD15 0004177 62700AD 6/27/00 7.48 6.50 19560FJ70,G4-1.6MKD-GD15 0004178 062700AD 6/27/00 14.9 ' 12.9 19561F,F0,G4-1.6MKD-GD15 0004179 062700AD 6/27/00 17.4 15.2 19562FJF0.G4-1.6MKD-GD15 0004180 . 062700AD 6/27/00 11.4 9.92 19563FF0.G4-1.6MKD-GD15 0004181 062700AD 6/27/00 8.95 7.78 19564FF0.G4-1.6MKD-GD15 0004182 062700AD 6/27/00 17.3 15.1 AVERAGE: 12.7 11.1 STANDARD DEVIATION: 3.77 3.28 3M Environmental LaDoraiory Amended Page 35 of 103 Page 370 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 . LRN-U2849 ! Centre Study No.: 023-016 : ' Sponsor Protocol No.: FACT-TOX-110 Table XVI. Summary of PFOS residues in G5-3.2MKD on GD15 Sponsor Centre ID Set Corrected Date Residue Found Residue Found ID Number Extracted 19565FJF0.G5-3.2MKD-GD15 0004183 062800AD 6/28/00 (p s/2 ) 35.2 (ps/2) 30.6 19566FF0.G5-3.2MKD-GD15 0004184 062800AD 6/28/00 15.2 13.2 19567F,F0,G5-3.2MKD-GD15 0004185 062800AD 6/28/00 20.4 17.7 19568FJFD,G5-3.2MKD-GD15 0004186 062800AD 6/28/00 46.9 40.8 19569F.F0.G5-3.2MKD-GD15 0004187 062800AD 6/28/00 36.3 31.6 19570F,F0,G5-3.2MKD-GD15 0004188 062800AD 6/28/00 31.1 27.0 1957IFF0.G5-3.2MKD-GD15 0004189 062800AD 6/28/00 28.4 24.7 19572FF0.G5-3.2MKD-GD15 0004190 062800AD 6/28/00 20.5 ' 17.9 19573FF0,G5-3.2MKD-GD15 0004191 062800AD 6/28/00 39.8 . 34.6 19574FF0,G5-3:2MKD-GD15 0004192 062800AD 6/28/00 47.3 41.1 19575FJD,G5-3.2MKD-GD15 0004193 062800AD 6/28/00 40.7 35.4 19577FJ0.G5-3.2MKD-GD15 0004194 062800AD 6/28/00 47.3 41.1 19579FJF0.G5-3.2MKD-GD15 0004195 062800AD 6/28/00 33.4 29.1 19580F.F0.G5-3.2MKD-GD15 0004196 062800AD 6/28/00 32.7 28.4 AVERAGE: 33.9 29.5 - STANDARD DEVIATION: 10.2 8.91 3M Environmental Laboratory Amended Page 36 of 103 Page 371 3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 CentreiStudy No.: 023-016 Sponsor Protocol No.: FACT-TOX-110 Table XVII. Summary of PFOS residues in G l-Control on GD21 Sponsor Centre BD Set Corrected Date Residue Found Residue Found ID Number Extracted 19504F,F0,Gl-Control-GD21 0004197 062900A 6/29/00 (ue/a) ND (ue/g) ND 19505F,F0,Gl-Control-GD21 0004198 062900A 6/29/00 ND ND 19506FFO,G1-Control-GD21 0004199 062900A 6/29/00 0.0123 0.0107 19507Fj70,Gl-Control-GD21 0004200 062900A 6/29/00 ND ND 19509FF0,Gl-Control-GD21 0004201 062900A 6/29/00 0.843 0.0733 19512FJ*0,G1-Control-GD21 0004202 062900A 6/29/00 0.0215 0.0187 19516FJO,G1-Control-GD21 0004203 062900A 6/29/00 <0.0100 < 0.00869 AVERAGE: 0.0183 0.0159 STANDARD DEVIATION: 0.0302 0.0263 ND = Not Detected and NQ = Not Quantifiable ' Note: The average and standard deviation were calculated by using 0.0100 as the value for those samples reported as < 0.0100,0.00869 for those reported as < 0.00869, and zero for those reported as ND or NQ. ' Table XVIII. Summary of PFOS residues in G2-0.1MKD on GD21 Sponsor Centre ID Set Corrected Date Residue Found Residue Found BD Number Extracted . (pg/g) 19518F,F0,G2-0.1MKD-GD21 0004204 0629-3000AD 6/29/00 0.735 (pgAr) 0.639 19519F,F0,G2-O.1MKD-GD21 0004205 062900A . 6/29/00 19521FF0.G2-0.1MKD-GD21 0004206 062900A 6/29/00 0.473 0.403 0.411 0.351 19523FJ0.G2-0.1MKD-GD21 0004207 062900A 6/29/00 0.402 0.349 19524FF0.G2-0.1MKD-GD21 0004208 062900A 6/29/00 0.494 0.429 19528FJ?0,G2-0.1MKD-GD21 0004209 062900A 6/29/00. 0.456 0.396 19529FJ0.G2-0.1MKD-GD21 0004210 062900A 6/29/00 0.391 0.340 AVERAGE: 0.479 0.417 STANDARD DEVIATION: 0.120 0.104 3M Environmental Laboratory Amended Page 37 of 103 Page 372 3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 - - Centre Study No.: 023-016 , Sponsor Protocol No.: FACT-TOX-110 Table XIX. Summary of PFOS residues in G3-0.4MKD on GD21 Sponsor Centre ID Set Corrected Date Residue Found Residue Found ID Number Extracted (gg/g) (pg/g) 19533F.F0.G3-O.4MKD-GD21 0004211 0629-3000AD 6/30/00 1.68 1.46 19534FJ0.G3-0.4MKD-GD21 0004212 0629-3000AD 6/30/00 3.10 2.70 19536F.F0.G3-O.4MKD-GD21 0004213 0629-3000AD 6/30/00 1.85 1.61 19543FJ0,G3-O.4MKD-GD21 0004214 0629-3000AD 6/30/00 2.23 1.94 19546F.F0.G3-0.4MKD-GD21 0004215 0629-3000AD 6/30/00 2.23 1.94 19548FJ0.G3-O.4MKD-GD21 0004216 0629-3000AD 6/30/00 5.42 4.71. AVERAGE: 2.75 2.39 STANDARD DEVIATION: 1.39 1.21 Table XX. Summary of PFOS residues in G4-1.6MKD on GD21 Sponsor Centre ID Set Corrected Date ;Residue Found Residue Found ID Number Extracted _ (pg/g) (pg/g) 19556FJUG4-1.6MKD-GD21 0004217 0629-3000AD 6/30/00 18.3 15.9 19559F,F0,G4-1.6MKD-GD21 0004218 0629-3000AD 6/30/00 10.6 9.19 19560FF0.G4-1.6MKD-GD21 0004219 0629-3000AD 6/30/00 11.2 9.72 : 19562FJ0.G4-1.6MKD-GD21 0004220 0629-3000AD 6/30/00 5.68 4.94 AVERAGE: 11.4 9.93 STANDARD DEVIATION: 5.19 4.51 Table XXI. Summary of PFOS residues in G5-3.2MKD on GD21 Sponsor Centre ID Set Corrected Date Residue Found Residue Found ID Number Extracted _ (pg/g) (pg/g) 19568FJ0.G5-3.2MKD-GD21 0004221 0629-3000AD 6/30/00 25.1 21.8 . 19569FJF0.G5-3.2MKD-GD21 0004222 0629-3000AD 6/30/00 21.7 18.8 1957IFJ0,G5-3.2MKD-GD21 0004223 0629-3000AD 6/30/00 21.5 18.7 19572F,F0,G5-3.2MKD-GD21 0004224 0629-3000AD 6/30/00 15.0 13.0 19577F.F0.G5-3.2MKD-GD21 0004225 0629-3000AD 6/30/00 27.1 23.5 19579FJF0.G5-3.2MKD-GD21 0004226 0629-3000AD 6/30/00 28.3 24.6 AVERAGE: 23.1 20.1 STANDARD DEVIATION: 4.83 4220 Amended Page 38 of 103 3M Medical Department Study: T-6295.12 Appendix H: Interim Certificate of Analyses Analytical Report: FACT TOX-110 LRN-U2849 3M Environmental Laboratory Page 374 3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 Centre Analytical Laboratories, Inc. 3048 Research Drive Phone: (814) 231-8032 State College, PA 16801 Fax: (814) 231-1253 or (814) 231-1580 INTERIM CERTIFICATE OF ANALYSIS Revision 1(9/7/00) Centre Analytical Laboratories COA Reference #: 023-018B 3M Product: PFOS,Lotl71 Reference#: SD-009 ____________________ Purity: 86.4%____________________ Test Name Specifications Purity1 Result 86.4% Appearance White Crystalline Powder Conforms Identification NMR Positive Metals (ICP/MS) 1. Calcium 1. 0.017 wt.Avt.% 2. Magnesium 2. 0.007 wt./wt.% 3. Sodium 3. 1.355 wt./wt.% 4. Potassium2 4. 6.552 wt/wt.% 5. Nickel 5. 0.003 wt./wt.% 6. Iron 6. 0.004 wt./wt.% 7. Manganese 7. <0.001 wt.Avt.% Total % Impurity (NMR) 1.00 wt.Avt.% Total % Impurity 10.60 wt./wt.% (LC/MS) ( - Total % Impurity None Detected (GC/MS) Related Compounds - POAA 0.30 wt.Avt.% Residual Solvents (TGA) None Detected Purity by DSC Not Applicable3 Inorganic Anions (IC) 1. Chloride 1. <0.015 wt.Avt.% 2. Fluoride 2. 0.27 wt.Avt.% 3. Bromide 3. <0.040 wt./wt.% 4. Nitrate 4. <0.009 wt./wt.% 5. Nitrite 5. <0.006 wt.Avt.% 6. Phosphate 6. <0.007 wt.Avt.% 7. Sulfate4 7. 8.82 wt.Avt.% Organic Acids5(IC) 1. TFA 1. <0.1 wt.Avt.% 2. PFPA 2. <0.1 wt.Avt.% 3. HFBA 3. <0.1 wt.Avt.% 4. NFPA 4. <0.25 wt.Avt.% Elemental Analysis': 1. Carbon 1. Theoretical Value = 17.8% 1. 12.08 wt.Avt.% 2. Hydrogen 2. Theoretical Value = 0% 2. 0.794 wt.Avt.% 3. Nitrogen 3. Theoretical Value = 0% 3. 1.61 wtVwt.% 4. Sulfur V 5. Fluorine 4. Theoretical Value = 5.95% 5. Theoretical Value = 60% 4. 10.1 wt.Avt.% 5. 50.4 wt.Avt.% COA023-018B Page 1 o f3 3M Environmental Laboratory Page 375 3M Medical Department Study: T-6295.12 Analytical Report: FACTTOX-110 LRN-U2849 Centre Analytical Laboratories, Inc. 3048 Research Drive Phone: (814) 231-8032 State College, PA 16801 Fax: (814) 231-1253 or (814) 231-1580 INTERIM CERTIFICATE OF ANALYSIS C entre Analytical Laboratories COA Reference #: 023-018B Date o f Last Analysis: 08/31/00 Expiration Date: 08/31/01 Storage Conditions: Frozen <-10C Re-assessment Date: 08/31/01 'Purity = 100% - (sum o f metal impurities, 1.39% +LC/MS impurities, 10.60%+Inorganic Fluoride, 0.27%+NMR impurities, 1.00%+ POAA, 0.30%) Total impurity from all tests = 13.56% Purity = 100% - 13.56% = 86.4% 2Potassium is expected in this salt form and is therefore not considered an impurity. 3Purity by DSC is generally not applicable to materials of low purity. No endotherm was observed for this sample. 4Sulfur in the sample appears to be converted to SO4 and hence detected using the inorganic anion method conditions. The anion result agrees well with the sulfur determination in the elemental analysis, lending confidence to this interpretation. Based on the results, the SO4 is not considered an impurity. 5TFA HFBA NFPA PFPA Trifluoroacetic acid Heptafluorobutyric acid Nonofluoropentanoic acid Pcntafluoropropanoic acid 6Theoretical value calculations based on the empirical formula, CsFnSOj'K* (MW=538) This work was conducted under EPA Good Laboratory Practice Standards (40 CFR 160). COA023-018B 3M Environmental Laboratory Page 2 o f3 Page 376 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 --------------------------- CKN-U2849 Centre Analytical Laboratories, Inc. 3048 Research Drive Phone: (814) 231-8032 State College, PA 16801 Fax: (814) 231-1253 or (814) 231-1580 INTERIM CERTIFICATE OF ANALYSIS C entre Analytical Laboratories COA Reference #: 023-018B LC/MS Purity Profile: Im purity C4 C5 C6 C7 T o ta l wt7wt. % 1.03 1.56 6.38 1.63 10.60 Note: The C4 and C6 values were calculated using the C4 and C6 standard calibration curves, respectively. The C5 value was calculated using the average response factors from the C4 and C6 standard curves. Likewise, the C7 value was calculated using the average response factors from the C6 and C8 standard curves. ( Prepared By: Datnd S. Bell Date Scientist, Centro'Aniilytical Laboratories // Reviewed By: (j* k fl) yiohn Flaherty ' Date / Laboratory Manager, Centre Analytical Laboratories V. COA023-018B Page 3 o f3 3M Environmental Laboratory Page 377 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Analytical Laboratories, Inc. 3048 Research Drive State College, PA 16801 Phone: (814) 231-8032 Fax: (814) 231-1253 or (814) 231-1580 (" INTERIM CERTIFICATE OF ANALYSIS Revision 1(9/7/00) Centre Analytical Laboratories COA Reference #: 023-018A 3M Product: PFOS,Lot217 Reference#: SD-018 ____________________ Purity: 86.9%______________ Test Name Specifications Result Purity1 86.9% Appearance White Crystalline Powder Conforms Identification NMR Positive Metals (ICP/MS) 1. Calcium 1. 0.005 wt./wt.% 2. Magnesium 2. 0.001 wt./wt.% 3. Sodium 3. 1.439 wt./wt.% 4. Potassium2 4. 6.849 wt./wt.% 5. Nickel 5. <0.001 wt./wt.% 6. Iron 6. 0.005 wt./wt.% 7. Manganese 7. <0.001 wt7wt.% Total % Impurity (NMR) 1.93 wt7wt.% Total % Impurity 8.41 wtVwt.% (LC/MS) I . Total % Impurity None Detected (GC/MS) Related Compounds - POAA 0.33 wt./wt.% Residual Solvents (TGA) None Detected Purity by DSC Not Applicable-1 Inorganic Anions (IC) 1. Chloride 1. <0.015 wt./wt.% 2. F luoride 2. 0.59 wt./wt.% 3. Bromide 3. <0.040 wt./wt.% 4. Nitrate 4. <0.009 wt./wt.% 5. Nitrite 5. <0.006 wt./wt.% 6. Phosphate 6. <0.007 wt./wt.% 7. Sulfate4 7. 8.76 wt./wt.% Organic Acids *(IC) 1. TFA 1. <0.1 wt./wt.% 2. PFPA 2. <0.1 wt./wt.% 3. HFBA 3. 0.10 wt./wt.% 4. NFPA 4. 0.28 wt./wt.% Elemental Analysis": 1. Carbon 1. Theoretical Value = 17.8% 1. 12.48 wtVwt.% 2. Hydrogen 2. Theoretical Value = 0% 2. 0.244 wt./wt.% 3. Nitrogen 3. Theoretical Value = 0% 3. 1.74 wt./wt.% 4. Sulfur 4. Theoretical Value = 5.95% 4. 8.84 wt./wt.% 5. Fluorine 5. Theoretical Value = 60% 5. 54.1 wt./wt.% COA023-018A Page 1 o f3 3M Environmental Laboratory Page 378 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Analytical Laboratories, Inc. 3048 Research Drive Phone: (814) 231-8032 State College, PA 16801 Fax: (814) 231-1253 or (814) 231-1580 INTERIM CERTIFICATE OF ANALYSIS Centre Analytical Laboratories COA Reference #: 023-018A Date o f Last Analysis: 08/31/00 Expiration Date: 08/31/01 Storage Conditions: Frozen <-10C Re-assessment Date: 08/31/01 'Purity = 100% - (sum o f metal impurities, 1.45% +LC/MS impurities, 8.41%+Inorganic Fluoride, 0.59%+NMR impurities, 1.93%+organic acid impurities, 0.38%+POAA, 0.33%) Total impurity from all tests = 13.09% Purity = 100% -13.09% = 86.9% 2Potassium is expected in this salt form and is therefore not considered an impurity. 3Purity by DSC is generally not applicable to materials of low purity. No endotherm was observed for this sample. 4Sulfur in the sample appears to be converted to SO4 and hence detected using the inorganic anion method conditions. The anion result agrees well with the sulfur determination in the elemental analysis, lending confidence to this interpretation. Based on the results, the SO4 is not considered an impurity. 5TFA HFBA NFPA PFPA Trifluoroacetic acid Heptafluorobutyric acid Nonofluoropentanoic acid Pentafluoropropanoic acid 6Theoretical value calculations based on the empirical formula, CgF|7S0 3 'K+(MW=538) This work was conducted under EPA Good Laboratory Practice Standards (40 CFR 160). COA023-018A 3M Environmental Laboratory Page 2 o f3 Page 379 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Analytical Laboratories, Inc. 3048 Research Drive Phone: (814) 231-8032 State College, PA 16801 Fax: (814) 231-1253 or (814) 231-1580 INTERIM CERTIFICATE OF ANALYSIS C entre Analytical Laboratories COA Reference #: 023-018A LC/MS Purity Profile: Im purity C4 C5 C6 C7 T o ta l w t./w t % 1.22 1.33 4.72 1.14 8.41 Note: The C4 and C6 values were calculated using the C4 and C6 standard calibration curves, respectively. The C5 value was calculated using the average response factors from the C4 and C6 standard curves. Likewise, the C7 value was calculated using the average response factors from the C6 and C8 standard curves. I Prepared By: y /, f" Q David S. Bell Date Scientist, Centre Analytical Laboratories // Reviewed By: fh /John Flaherty 9 Date Laboratory Manager, Centre Analytical Laboratories COA023-018A 3M Environmental Laboratory Page 3 of 3 ' Page 380 3M Medical Department Study: T-6295.12 Appendix i: Report Signature Page Analytical Report: FACT TOX-110 LRN-U2849 Marvin T. Case, D.V.M., Ph.D., Study Director 0ate John L. Butenhoff, Ph.D., Sponsor Representative . --" W illiam Reagen, Ph.D., Laboratory Manager r /y /* ' Date S 'y k s /o / Date ____ 7 ^ -- r.r -- ___________________________________________ Kris 3. Hansen, Ph.D., P rinciple A nalytical Investigator >5 /o ' J J n r Date 3M Environmental Laboratory Page 381