Document NGB2waa1a8m458j3977G4MdQ

3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Table 7: Ambient-Temperature Stability of PFOS in Rat Serum After 24 Hours N A Not Applicable RSD = (SD/Mean)x 100 %DEV fkom 0Hour = [(X Hour Conc.- 0 Hour Conc.)/O Hour Conc.]x 100 3M Environmental Laboratory 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 Rat Serum at -20 O C L Nominal 0.209 PFOS (ug/mL) I I 7 Days 13Days 33 Days I I (Run 6)* (Run3)** (Run 16) 0.22:5 0.29!2 0.211 i 0.245 0.210 0.234 (0.2 ug/mL) 0.208 0.21 8 0.217 0.228 0.208 0.21 9 0.214 0.232 0.207 0.209 0.2211 0.213 0.235 RSD (%) I QC 2 5.71 (6ug/mL) 6.00 6.06 -1.32 5.7 6.2 5.69 I Mean 5.89 RSD(%) I 3.08 %DEV from 0 Hour NA 20.2 QC 3 18.6 (18 18.9 18.2 18.9 Mean 18.9 ; RSD (%) 3.96 NA N A Not Applicable RSD = (SD/Mean) x 100 %DEV fiom 0 Hour = [(Conc. - 0 Hour Conc.)/O Hour Conc.] x 100 *FromProtocol FACT-TOX-110 **From Protocol FACT-TOX-111 3M Environmental Laboratory ADVANCED BIOANALYTICAL SERVICES, INC. Page 27 of 50 99VDJAO1.MLDOC Page 198 3NI Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Table 9: Stabilityof PFOS in Rat Serum After Three Freeze/Thaw Cycles PFOS (pg/mL) icle 3 Cycles 87 0.177 NA:Not Applicable RSD = (SDMean) x 100 %DEV f?om 0 Cycle = [(3 Cycle - 0 Cycle)/O Cycle] x 100 3M EnvironmentalLaboratory Page 28 of50 99VDJAOI.MI.DOC Page 199 3NI Medical Department Study: T-6295.12 i Analytical Report: FACT TOX-110 LRN-U2849 Table 10: Reproducibility of Reinjecting Extracted Samples Containing PFOS after 27 Hours in Reconstitution Solution (0.2 P d d ) Qcl Mean I I PFOS (pg/mL) t=o Hours t=27Hours (RUII 10) (Run 12) I 0.187 0.1 86 0.1 82 0.1 89 0.1 82 0.187 0.182 0.1.88 0.188 0.190 0.1 85 0.187 6.44 6.02 6.32 6.19 6.19 6.55 RSD (%) RE (%) N A Not Applicable RSD = (SDMean) x 100 RE = [(MeanRheoretical)-l] x 100 17.3 17.5 1s.o 1'7.4 -3.29 18.2 18.1 17.3 . 17.6 2.40 -1.73 3NI EnvironmentalLaboratory ADVANCED BIOANALYTICAL SERVICES. INC. Page 29 of 50 99VDJA01.MI.DOC Page 200 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Table 11: Extraction Recovery of PFOS .from Rat Serum & Theoretical Conc. I - Pre-Extraction in serum (pg/mL) Response (Area Ratio) PFOS 0.03160 k s Post-Extraction ponse (Area Ratio) I % Recovery* - L .649 .679 1.857 90.4 ,719 1.869 92.0 .s35 1.887 81.4 .612 Mean I 1.639 Tetra-H-PFOS 0.4068 Spike IS 0.3884 (4 I.rg/mL) Run 13 0.4043 0.3921 0.4078 *(IndividualResponse for Pre-Ext./Individual Response for Post-Ext.) x 100 3NI Environmental Laboratory ADVANCED BIDANALYTICAL SERVICES, INC. Page 30 of 50 99VDJAO1 .MI.DOC Page 201 3NI 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 98 - 91 84 77 70 - 63 - 56 - 49 - 42 - 35 - 28 21 - 14 - . I 399 360 420 dz 3.19e7 cps 499 [M-K]- -480 540 3NI EnvironmentalLaboratory ADVANCED BlOANALYTlCAL SERVICES, INC. Page 31 of 50 99VDJAO1 .MLDOC Page 202 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Figure 2: Full-Scan Single MS Mass Spectrum of Tetra-H-PFOS Spectrum from Tetra-H-PFOS Q1 1.94e7 cps 27 M-HI- I 350 400 45 0 dz 3" Environmental Laboratory ADVANCED BiOANALYTICAL SERVICES, INC. Page 32 of 50 99VDJAO 1.MI.DOC Page 203 3Nl Medical Department Study: T-6295.12 I Analytical Report: FACT TOX-110 LRN-U2849 Figure 3: Mass Chromatograms of PFO!; and its Internal Standard in Control Blank Rat Serum Extract d z =427.0 1.07e3 cps I I 1 m/z =499.0 I I 2 3 Time, min I 4 3.54e2 cps I I 1 2 3 4 Tim:, min All quantitation results and figures were prepared from non-smoothed data. Analvte Qa etcnh.on Ti.me PFOS d z = 499.0 3.1 Tetra-H-PFOS (IS) m h = 427.0 2.9 3NI Environmental Laboratory ADVANCED EIOANALYTICAL SERVICES, INC. Page 33 of 50 99VDJA01.MI.DOC Page 204 3hl 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) mlz =427.0 2.34e5 cps I I m/z =499.0 I I 2 3 Time, min 3.78e2 cps J III I 1 2 3 4 Timi:, min All quantitation results and figures were prepared from non-smoothed data. Anal* PFOS Tetra-H-PFOS(IS) len m/z = 499.0 mlz = 427.0 Retention 3.1 2.9 3M Environmental Laboratory 99VDJAO1.MI.DOC Page 205 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Figure 5: Mass Chromatograms of Calibration Standard 1 in Rat Serum Extract Containing PFOS (0.05 pglmL) and the Internal Standard mlz =427.0 3.82e5 cps 1 mlz =499.0 2 3 Time, min 4 2.1 le3 cps 1 2 3 4 Time, min All quantitationresults and figureswere prepared from non-smootheddata. Analvte PFOS Tetra-H-PFOS (IS) h3.I m/z = 499.0 d z =427.0 Betention Time 3.1 2.9 3NI Environmental Laboratory ADVANCED BIOANALYTICAL SERVICES, INC. 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 1 d z =499.0 2 3 Time, min 4 5.14~5cps 1 2 3 4 Timi:, min All quantitation results and figures were prepared from non-smoothed data. Analvte PFOS Tetra-H-PFOS (IS) - Ion mlz = 499.0 d z = 427.0 Retention Time 3.1 2.9 3MI EnvironmentalLaboratory ADVANCED BIDANALYTICAL SERVICES, INC. Page 36 of 50 99VDJA01 .MI.DOC Page 207 3M 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.l.O A.2.0 A.3.0 A.4.0 A.5.0 A.6.0 A.7.0 A.8.0 A.9.0 A.lO.O A.ll.O Chemical Structure(s)........................................................................................ 38 Specimen(s)....................................................................................................... 38 Assay Principle.................................................................................................. 38 Compounds......;................................................................................................ .38 Chemicals .....................................................................................................3..8... Materials and Equipment..............................................................................-.3..9. Preparation of Solutions .................................................................................... 42 Preparationof Quality Control Samples............................................................45 Liquid-liquidExtraction Procedure .................................................................. .46 Instrument Conditions ........................................................................................ 47 Calculations..................................................................................................... ..49 3MI Environmental Laboratory ADVANCED BIOANALYTICAL SERVICES, 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.l.O CHEMICAL STRUCTURE(S) 0 II c~F,,-s-o-K+ II 0 Potassium Perfluorooctanesulfonate MW=538 C6F 13- 0 C -C- II 1S1-OH H2 H2 0 IH,lH, 2H,2H-PerfluorooctaneSulfonicAcid MW =428, Internal Standard A.2.0 SPECIMEN(S) This assay uses 50-pL aliquots of rat serum. Rit serum samples are stored at -20 O 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 (LCh4S) 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, Ihc., Minneapolis, MN. A.5.0 CHEMICALS All chemicals may be substituted with that of an equivalent manufacturer and grade of chemical. 3NI EnvironmentalLaboratory ADVANCED BlOANALYTICAL SERVICES, INC. Page 38 of 50 99VDJAOl .MI.DOC Page 209 3M Medical Department Study: T-6295.12 I Acetonitrile Ammonium Aceiate Ethyl Acetate Methanol Rhodapex mass calibration solution Sodium Bicarbonate Sodium Carbonate, Annhydrous TetrabutylammoniumHydrogen Sulfate Water Rat Serum Rat Serum Analytical Report: FACT TOX-110 LRN-U2849 Cat# 015-4, Burdick & Jackson, Muskegon, MI 49442 Cat# :!4,019-2, Aldrich, Milwaukee, WI 53201 Cat# 'I00-4, Burdick & Jackson, Muskegon, MI 49442 Cat# 230-4, Burdick & Jackson, Muskegon, MI 49442 A mixture of 0.11 mM trifluoroacetic acid (J.T.13aker, Phillipsburg, NJ 08865),0.52 mMIthodapex CO-436,l mM ABEX EP110 and 1 mM ABEX EP-120 (all three supplied by Rhone-Poulenc, Inc., Cmbury, NJ 08512) in 1:l rneth;mol:water. Cat# 3509-01, J.T. Baker, Phillipsburg, NJ 08865 Cat# 3604-01, J.T. Baker, Phillipsburg, NJ 08865 HPLC Grade, Cat# 5360-07, J.T. Baker, Philljpsburg, NJ 08865 High Purity (NANOpure), Barnstead Modd D7331 Ultrapure Water System, Dubuque, IA 52001 Lmpire Biological Laboratories, Piperville, PA 18947 Harlan Bioproducts for Science, Inc., Indianapolis, IN 46229 A.6.0 MATERIALS AND EQUIPMENT Mass Spectrometer PE SCIEX API 365 atmospheric pressure ionization tandem triple quadrupole mass spectrometer equipped with TurboIonSprayTMinterface, PE SCIEX,Concord, Ontario LAK 4V8 Data system API Standard Sofiware,MacQuan v 1.4,LC2Tune v 1.3, MacDAD v 1.3, Bundler v 1.3, Multiview v 1.3, ADVANCED BIOANALYTICAL SERVICES, INC. Page 39 of 50 99MJA01 .MI.DOC 3NI Environmental Laboratory Page 210 3V1 Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 HPLC Pump LC Pump Controller Auto sampler HPLC Column Harvard syringe P U P 11 Multi-tube vortexer Balance pH Meter Mechanical shaker NANOpure-W water purification system Microbalance Micro weigh boats Weigh boats Sor~alRl T-6OOOD refrigerated centrifuge Beckman GS6KR Refrigerated centrifuge and Sample Control v 1.3, on a Power Macintosh, PE SCEX, Concord, Clntario L4K 4V8 Shimadzu LC-10A.I) pump, Shimadzu Co., Columbia, MD 21046 Shimadzu SCL-lo!,, Shimadzu Co., Columbia, MD 2 1046 Waters 717plus,Witers Associates,Millipore Corporation, Milford, MA 01757 Betasil C,8,5 pm piuticle 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, AND Ltd., Tokyo, Japan Model 340, Coming Inc., Corning, NY 14830 Cat# 6000, Eberbach Corp., Ann Arbor, MI 48106 Barnstead, Dubuque, IA 52001 Model MT-5, Metfler-Toledo Inc., Hightstown, NJ 08520 Cat# 0219-0041, Pdcin-Elmer Corp., Norwalk, CT 06859 Cat# 12577-025,VWR Scientific,West Chester, PA 19380 Cat# 83071, DuF'oiit Co., Wilmington, DE 19898 Beckman, Palo Alto, CA 94304 3Nl EnvironmentalLaboratory ADVANCED BlOANALYTlCAL SERVICES, INC. Page 40 of50 99VDJA01.MI.DOC Page 21 1 3RE 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, Zymarlr 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, (2-25, CP-50, CP-250, Rainin Instrument Co., Wobum, MA 01888 13 mm x 100mm, Cat# 15070-574, V W R Scientific, West Chester, PA 'I9380 16.5 x 57 mm, Cat# 60.542, Sarstedt, Inc., Newton, SC 28658 16.5 x 101mm, Cat# 60.541, Sarstedt, Inc., Newton, SC 28658 Cat# 78-127-0019-100, Elkay Products, Inc., Worcester, MA 01607 Cat# 28014P-10,233014P-25,28014P-100,28014P1000, JSimble/Konl.es, Vineland, NJ 08360 Cat# 953781-OOOO.,953751-0000,953753-0000, 953826-0000,953827-0000,KimbleKontes, Vineland, NJ 08360 0.45 pm Scientific Resources Inc., Cat# 74547-NN, Eatontown, NJ 075'24 0.005'' id. x 1/16"o.d., Cat# 1535,Upchurch Scientific Inc., Oak Harbor, WA 98277 Supplied in-housefrom bulk tank, BOC Gasses, Buffalo, NY 14210-2005 Cat# 1750, Hamilton Company, Reno,NV 89510 Other general laboratory glassware and supplies were used. 3CA Environmental Laboratory ADVANCED BlOANALYTICAL SERVICES, INC. Page 41 of 50 99VDJAOl .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 "Cand bring to ambient temperature before we. Standard Spiking Solutions Prepare Standard Working SolutionsA 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 (PL 1 1000 800 400 200 100 250 125 62.5 500 250 Solution Used Stock Stock Stock Stock Stock 200 p g h L (A) 200 p.g/nL (A) 200 pg/nL (A) 10 PdmL 10 Pg/KL (F) PFOS Final Cone. (I.rg/W 200 160 80 40 20 10 5 2.5 1.0 0.5 3NI Environmental Laboratory ADVANCED BlOANALYTlCAL SERVICES, INC. Page42 of 50 99VDJA01 .MLDOC Page 213 3Nl 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 1mg/mL sohtion. 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 StandardWorking 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 pig/& Tetra-H-PFOS solution. Store the solution at 4 "Cand bring to ambient temperature before use. Prepare fresh solution as needed. A.7.4 Preparation of Standard Curve and Control Blank Prepare fiesh calibration standards for each analytical runby combining 360 pL of rat control serum and 40 pL of the analytical Stimdard 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. 3NI Environmental Laboratory ADVANCED BIDANALYTICAL SERVICES, INC Page 43 of 50 99VDJAOI .MI.DOC Page 214 3M Medical Department Study: T-6295.12 I Analytical Report: FACT TOX-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 3 60 3 60 360 360 360 360 360 360 3 60 360 Final Conc. (PdmL) 20 16 8 4 2 1 0.5 0.2s 0.1 0.05 A.7.5 Preparation of Other Solutions 0.25 M Sodium Carbonate/0.25 M Sodium Iiicarbonate: 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-mLvolumetric 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 amm.onium acetate to a 100-mLvolumetric flask, dissolve in NANOpure water, and stir until completely dissolved. Dilute to the mark with NANOpure water. Store at ambient temperature. Prepare a fiesh solution every three months. 2 mM Ammonium Acetate: Combine 2 mL, of 1M ammonium acetate and 900 d NANOpure water in a 1000-mLvolumetric flisk; stir until completely mixed. Dilute to the mark with NANOpure water. Store at ambient temperature. Prepare a fresh solution every three months. 3FA Environmental Laboratory ADVANCED BlOANALYTlC AL SERVICES.INC. Page 44 of 50 99VDJA01 .MI.DOC Page 215 3Nl 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 1:emperature. Prepare a fresh solution every three months. 1 M Sodium Hydroxide: Add 10mL 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 TetrabutylammoniumHydrogen Sulfate, pH 10: Weigh 16.98 g of tetrabutylammoniumhydrogen sulfate and dissolve in 40 rnL of NANOpure water. Adjusted the pH to 10.0with 10M and 1M sodium hydroxide. Transfer the solution to a 100-mLvolumetric flask and dilute to the inark with NANOpure water. 10:90 Methanol:2 mM Ammonium Acetate (Mobile Phase Eluent A): Add 100 mL of methanol and 900 mL of 2 mMammonium acetate to a 1000-mLPyrex 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:lO Methanokl: mM Ammonium Acetate (Mobile Phase Eluent B): Add 900 mL of methanol and 100mL of 2 m Mammonium acetate to a 1000-mLPyrex 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 fiesh solution every three months. A.8.0 PREPARATION OF QUALJTY 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 3NI Environmental Laboratory ADVANCED BlOANALYTlCAL SERVICES, INC. Page 45 of 50 99VDJA01.MI.DOC Page 2 16 3M Medical Department Study: T-6295.12 i Analytical Report: FACT TOX-110 LRN-U2849 with the appropriate volume of methanol to yield a 1mg/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 p g / d PFOS): Add 1000 pL QC Stock Solutionto a 10mL class "A" volumetric flask to yield a IOO-pg/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 at -20 "C:. QC3 (18 &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 h z e n at -20 "C. QC2 (6 pg/mL PFOS): Add 150 pL of QC Stack Solution to a 25-mL class "A" volumetric flask to yield a 6-pghL solution. Dilute to the mark with control serum, cap and mix. Aliquots of approximately 0.5 ml, are placed in polypropylene vials and stored h z e n at -20 "C. QCl (0.2 pglmL PFOS): Add 111.1 pL of QC!3to 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 PIROCEDURE 1. Prepare fiesh calibrationstandards for each.run according to Section A.7.4. 2. Thaw QC samples. 3. Prepare QC4 samples(diluted 1in 10)in two steps: a) Add 180 pL of control senun and 20 pL of QC4 into 1.7-mT, microtubes and 3NI Environmental Laboratory ADVANCED BlOANALYTlCAL SERVICES. INC. Page 46 of 50 99VDJAOI .MI.MX: Page 21 7 3Nl Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 vortex. b) Aliquot 50 pL of diluted QC4 into labeled 16x 100mm polypropylene tubes. 4. Aliquot 50 pL of each control blank, zero sample, calibration standard, and quality control samples 1-3 into labeled 16 r: 100 mm polypropylene tubes. Standards and blanks are analyzed in duplicate. All QC samples are analyzed in replicates of five. 5. Add 1mL of 0.5 M tetrabutylammonium hydrogen sulfate, pH 10 to each tube. 6. Add 2 mL. of 0.25 M sodium carbonaWO.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 p L water). 8. Add 5 mL ethyl acetate to each tube. 9. Shake the tubes on a reciprocal shaker at mcdium speed for 20 minutes. 10. Centrifugethe 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 acetatelayer to a fresh, labeled 13x 100mm polypropylene tube. . 13. Evaporate the ethyl acetate layer to dryness in a TurboVap* at approximately 20 "C under nitrogen. 14. Reconstitute the dried extracts in two steps: Add 500 pL acetonitrileand vohex for 60 seconds. Then add 500 pL water and vortex for 60 seconds. 15. Transfer 200 CCT, of the extract to a labeled polypropylene autosampler vial. k l O . O INSTRUMENT CONDITIONS HPLC Conditions: Eluent (gradient) Mobile Phase A: 10:90 methanol:2 mM ammonium acetate (v/v) Mobile Phase E:9O:lO methanol:2 mM ammonium acetate (v/v) 31vI Environmental Laboratory Page 47 of 50 99VDJAO 1 .MI.DOC Pagem 3NI Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Eluent Gradient Conditions: Time (min) 0 1 .o 5 5.5 8 Flow Rate Autosampler Autosampler Temperature Injection Volume HPLC Column HPLC Column Temperature Typical Initial Column Pressure Autosampler Run Time %B 100 45% stop 300 p:Umin. Waters 717plus Ambient 3to5pL Keystone Betasil C,8,5 pm particle size, 2.1 mmx50mm Ambient 90 bar 8.0 minutes Analvte Rmresentative Retention Time (minutes) PFOS Tetra-H-PFOS (IS) 1 3.1 2.9 Curtain Gas Nebulizer Gas TurboIonSpraym Temperature TurboIonSpraym Auxiliary Gas UHF Nitrogen UHP Nitrogen 400 "C UHP Nitrogen at 8 L/min 3Nl Environmental Laboratory ADVANCED BIOANALYTICAL SERVICES, INC. Page 48 of 50 99VDJA01.MI.DOC Page-2t4 3bI 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 mlz =:427.0 - Dwell- Time 500 ms 500 ms Ionization Mode Ion Spray Voltage Declustenng Potential MS Acquisition Time Pause Time Negative Ion -3000 V -35 v 5 miniites 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 pWmin into a flow of 190 pLlmin 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 approximately0.6 amu at half-height. One set of calibration standardswas injected at the beginning and one set of calibration standards was injected at the end of each analytical run (tray). A . l l . O CALCULATIONS Calculated concentrationsare 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 fiom 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.S.3.1.01). All data were rounded to no less than three significant figures by ABS prior to reporting. ~ - __ _ - ADVANCED BlOANALYTICAL SERVICES, INC. Page 49 of 50 99VDJA01 .MI.DOC 3N1 Environmental Laboratory Page 220 31\11Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 AcceDtance Criteria for SamDle Analvsis Calibration standards: The coefficient of determination (?)must be 20.98. 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. Iff 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 BIOANALYTICAL SERVICES, INC. Page SO of SO 99VDJAdl .MI.DOC Page 221 3 M Medical Department Study: T-6295.12 ADVANCED BIOANALYTICAL SERVICES, INC. BIOANALYTICAL REPORT Analytical Report: FACT TOX-110 LRN-U2849 Title: Quantitative Determination of Perfluorooctanesulfonate (PFOS) in Rat Semn from Study #FACT-TOX-110Using Turbo Ion Spray LUMS c Date: Authors: 21 September 1999 David J. Anderson, 1M.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- I 10 Number of Pages: 24 Summary-and Conclusions The concentration of perfluorooctanesulfonate (PFOS) was determined in rat serum samples collected during the study #FACT-TOX-110using a sensitive, specific, accurate, and reproducible analytical method developed by AldvancedBioAnalytical Services, Inc., Ithaca, NY. Rat serum samples (50 pL) were extracted by a liquid-liquidextractionprocedure to isolate PFOS and the internal standard, IH, 1H,2H,2:H-perfluorooctanesulfonic acid (Tetra-H-PFOS). Following evaporation and reconstitution, sample extracts were analyzed by turbo ion spray liquid chromatographyhnass spectrometry(LCMS) in the negative ion mode. All samples were successhlly anaIyzed within six runs. The lower limit of quantitation was 0.05 p&/rnL.for PFOS. The precision of this assay (RSD) as determined from the c analysis of quality control samples for PFOS was 56.90%. The precision of this assay (RSD), as determined from the calibration standards for PFOS was 16.18%. The relative error (RE) of the assay, as determined from the analysis of the quality control samples ranged fiom 1.52 to 12.4%. The RE of the assay, as 'determinedfrom the analysis of calibration standards ranged from -4.62 to 10.3%. a a& 15 Catherwood Road Ithaca, New York 14850 (60172)66-0665 Fax (607)266-0749 3M Environmental Laboratory Page 222 3M Medical Department Study: T-6295.12 r-3 Analytical Report: FACT TOX-110 LRN-U2849 ADVANCED BIOANALYTICAL SERVICES SIGNATURE PAGE Title: Quantitative Determination of Perfluorooctanesulfonate (PFOS) in Rat Serum from Study #FACT-TOX-110 Using Turbo Ion Spray h LCMS ABS Report No.: 99 Reported by: Research ScieEtist 21 s ? q q Date Reviewed by: Authorized for Release by: Melissa Mapes, A S . Associate Auditor 9/SEP99 Date 21. JbQ..t/Y Date 3M EnvironmentalLaboratory Page 2 of 24 99ADJA02.MI.DOC Page 223 3M Medical DepartmentStudy: T-6295.12 n Analytical Report: FACT TOX-110 LRN-U2849 QAU STATEMENT Periodic inspectionsof the bioanalyticalportion of study #FACT-TOX-I 10 were conducted by the Quality Assurance Unit of Advanced BioAnalyticaI Services (ABS) for compliance with EPA GLP regulations (40 CFR Part 792). The study was inspected on the followingdates: A 25 June 1999; 6, 7,8,21 July 1999; 31 August 1999. Results of the inspectionwere reported to A B S M,anagementand Project Team Leader on : 25 June 1999; 6, 7,8,21 July 1999; 31 August 19!>9. Results of 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 2/@9j? Date 3 M Environmental Laboratory Page 3 of 24 99ADJAOZ.MI.DOC Page 224 3Nl Medical Department Study: T-6295.12 I h Analytical Report: FACT TOX-110 LRN-U2849 TABLE OF CONTENTS SIGNATUREPAGE ...........................................................................................................2 QAU STATEMENT............................................................................................................3 TABLE OF CONTENTS..........................................................................................,....,....4 LIST OF TABLES ............................................................................................................. 5 1. INTRODUCTION......................................................................................................6 1.1. Study Descriptionand 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 "4 6. REFERENCES ....................... ..................................................................................... 8 7. TABLES ...................................................................................................................... 9 ADVANCED EIOANALYTICAL 1.4 SERVICES, INC. 3NI EnvironmentalLaboratory Page 4 of 24 99ADJAOZ.MI.DOC Page 225 3Wl Medical Department Study: T-6295.12 I r Analytical Report: FACT TOX-110 LRN-U2849 LIST OF TABLES A Table 1: Summary of Sample and Assay Information for Protocol FACT-TOX-110...... 9 Table 2: Inter-Assay Precision and Accuracy for I'FOS Quality Control Samples in Rat Serum for Study #FACT-TOX-110.................................................................. 10 Table 3: Inter-Assay Precision and Accuracy for I'FOS Calibration Standards in Rat Serum for Study#FACT-TOX-110 .................................................................11 Table 4: CalibrationCurve Statisticsfor the Determination of PFOS in Rat Serum for Study #FACT-TOX-110 ..........................................................................,.......12 Table 5 : ConcentrationsofPFOS in Rat Serum Samples from Study #FACT-TOX-110 Following the 0 mgkg Dose.................. ......................................................... 13 Table 6: Concentrationsof PFOS in Rat Serum Samples fiom Study #FACT-TOX-110 Followingthe 0.1 mgkg Dose........................................................................ 16 Table 7: Concentrationsof PFOS in Rat Serum Samples fiom Study #FACT-TOX-110 Followingthe 0.4 mgkg Dose........................................................................18 Table 8: Concentrationsof PFOS in Rat Serum Sunples from Study #FACT-TOX-110 Following the 1.6mgkg Dose......................................................................... 20 Table 9: Concentrationsof PFOS in Rat Serum Samples from Study #FACT-TOX-110 Following the 3.2 mgkg Dose............................................................................ 22 Table 10:Repeat Analysis of Rat Serum Samples fi-omStudy #FACT-TOX-110 ..........24 . ADVANCED BIOANALYTICAL SERVICES, INC. 3M Environmental Laboratory Page 5 of 24 99ADJA02.MLDOC Page 226 ~______________ 3NI Medical Department Study: T-6295.12 I n 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-liquidextraction and turbo ion spray liquid chromatography/massspectrometry (LCMS) assay. 2. METHODS 2.1. Analytical Procedure(s) PFOS concentrationswere determined in rat serum according to the validated LCMS 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,1H,2H,2H-perfluorooctanesulfonic acid (Tetra-H-PFOS)fkom rat serum. Following evaporationto dryness and subsequent reconstitution,sample extracts were separatedby reversed-phasechromatography on a 2 x 50 mm (5 pm)Betasilm C,, column (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 (90:10methanol:2 mM ammonium acetate). PFOS concentrations were determined by turbo ion spray liquid chromatography/massspectrometry (LCNS) in selected ion monitoring (SIM)mode. The negative ions monitored were: m/z = 499.0 for PFOS [M-KIm/z =427.0 for Tetra-H-PFOS [M-HI` Study sample concentrationswere determined from a weighted (l/$), quadratic regression of peak area ratios (peak area of PFOS/peak area of Tetra-H-PFOS) versus nominal concentrationof ten calibrationstandards. The nominal concentrationsof the calibration standardswere 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 p g / d PFOS) were analyzed with each assay ba.tch in replicates of four. Dilution QC samples (QC4, 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 acceptancecriteria for calibration standards stipulated that the back-calculated concentrationsof 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 3NI Environmental Laboratory Page 6 of 24 99ADJA02.MLDOC Page 227 3Nl Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 concentration . The coefficient of determination (r') must be 20.98. The acceptance criteria followed are in accordancewith ABS Standard OperatingProcedures. A 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 e,achQC concentration must exhibit a deviation within f15%. A summary of the sample and assay information fclr study #FACT-TOX-110 is given in Table 1. 2.2. Assay Site All samples were analyzed at Advanced Biohalytical Services, Inc., Ithaca, NY. 2.3. Data Processing The data were collected using turbo ion spray LChAS 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 MacQuanwere saved as text files and uploaded to the A B S file server where a weighted (1/?) quadratic regression was performed using the software package Watson (v 5.3.1.01 PSS, Inc., Wayne, PA 19087). CXculations were performed on unrounded numbers. All calibration standard and QC 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 calibrationcurves 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 inaccuracyin the results from these standards;the relative error (RE) ranged between -4.62to 10.3%. ADVANCED BIDANALYTICAL SERVICES, INC. 3NI Environmental Laboratory Page 7 of 24 99ADJA02.MI.DOC Page 228 31vl Medical Department Study: T-6295.12 i c.l Analytical Report: FACT TOX-110 LRN-U2849 The slope, y-intercept, and coefficient of determination (9)for each analytical run are presented in Table 4. The rZ values ranged from 0.9955 to 0.9965 for PFOS in rat serum. A All ABS personnel assigned to analyze samples fox this project were required to successfully extract quality control (QC) samples end 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 concentrationsin rat serum from Study #FACT-TOX-110 are presented in Tables 5 through 9. All data were rounded to no less than thee significant figures before reporting in Tables 5 through 9. Study samples requiring repsat analysis for Study #FACT-TOX110 are presented in Table 10. 4. SUMMARY AND CONCLUSIONS The concentrationof PFOS was determined in rat serum samples collected during the study #FACT-TOX-110. PFOS was isolated from serum by a liquid-liquid extractionprocedure and determined by LC/MS. The quality of the determinations was satisfactorythroughout. The LLQ was 0.05 pg/mL PFOS. The precision of the assay, as determined fiom the analysis of quality control samples was 26.90%for PFOS. 5. DATA RETRIEVAL T h e calculated concentration data from the analysis of rat serum samples for Study #FACT-TOX-110are maintained on file in the archives of the Advanced BioAnalytical Services, Inc., Ithaca, NY in ABS Notebook 2324. 6. REFERENCES I- 1. Advanced BioAnalytical ServicesValidation 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 11999. ADVANCED BIOANALYTICAL c-r SERVICES, INC. 3M Environmental Laboratory Page 8 of 24 Page 229 3M -MedicalDepartment Study: T-6295.12 i a Analytical Report: FACT TOX-1I O LRN-U2849 7. TABLES Table 1: Summary of Sample and Assay Iinformation for Protocol FACT-TOX110 SpeciedMatrix: RatfSenun Sample Collection and Storage Information: AnticoagulantBtabilizer: Reported Sample CollectionDates: 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 1999to 8 July 1999 Analyte: Potassium perfluorooctanesulfonate (PFOS) Analytical Standard: Lot No: Source: PFOS 171 3M Internal Standard: Lot No: Source: 1H,1H,2H,2Hperfluorooctane sulfonic acid (Tetra-H-PFOS) 59909 3M Calibration Range: Regression Method: Weighting Factor: Lower Limit of Quantitation: Upper Limit of Quantitation: ADVANCED BIOANALYTICAL P. SERVICES. INC. 3M Environmental Laboratory Page 9 of 24 99ADJA02.MI.DOC Page 230 3hl Medical Department Study: T-6295.12 m 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 Run No. 4 5 6 7 Mean RSD (%) RE 1 PFOS Conceniratio (pg/mL) I I I - QCl QC2 QC3 QC4 0.2 I 6 1 18 I 100 QC4 100 I I QC4 100 I (1 in 10 D i l u t a (1 in 20 Dilution) (1 in 30 Dilution) 0.178 6.18 15.9 NA NA I NA 0.183 6.40 17.3 NA NA NA I I 1 0.177 6.32 17.3 NA NA NA - 0.187 I 6.32 I 17.3 I NA NA NA 0.210 I 6.24 I 17.9 I NA NA NA 0.215 5.79 17.2 NA NA NA 1 I 1 - 1 I 1 0.211 5.99 17.8 NA NA NA 0.211 5.87 17.9 NA NA NA 0.225 1 6.14 I 18.1 I 116* NA NA NA NA I I 0o.2221 6.1019 NA NA - NA NA NA I 1 - NA 0.214 0.225 6.42 18.7 6.22 I 19.1 NA NA 107 NA 114 NA 108 NA 0.222 6.15 19.5 NA NA 1 I I 1 0.218 6.39 19.8 NA 111 NA I I 1 0.219 6.27 19.3 NA 111 NA I 0.221 I 6.11 18.6 1 NA _L NA 106 0.212 6.15 19.2 NA 104 1 I I I 0.215 6.17 18.5 NA NA 100 I I I II - I 0.218 6.80 18.8 NA 0.212 I 6.21 1 18.3 I 112 6.90 3.17 4.84 2.37 12.4 I p RE = r(Mean-NorninalVNominal1 x 100% NA: Not applicable ' *Individualreplicate exceeds 15% acceptance criterion. ADVANCED EIOANALYTICAL P. SERVICES, INC. 3M EnvironmentalLaboratory Page 10 of 24 99ADJAO2.MLDOC Page 231 1 I 'i i P -. I, i .? .I .. Table 3: #InFtAerC-TA-sTsaOyXP-r1e1c0ision and Accuracy for PFOS Calibration Standardsin Rat Serum for Study ua RE = [(P;iean-Nominal)/Nominal]x 100% Page 11 of 24 A (D U 0 ;r 99ADJAOZ.MLDOC 2 0 31\41Medical Department Study: T-6295.12 I c Analytical Report: FACT TOX-110 LRN-U2849 Table 4: Calibration Curve Statistics for the Determination of PFOS in Rat Serum for Study #FACT-TOX-110 m y =Ax2+ Bx f C, weighted l/Y #- 3M EnvironmentalLaboratory Page 12 of 24 99ADJAOZ.MLDOC Page 233 ; i :i i ? P Table 5: Concentrations of PFOS in Rat Serum Samples from Study #FACT-TOX-110 Following the 0 mgkg Dose u a -cIn Sampling PFOS Concentration (pg/mL) N (D Time Rat Number* P 2 N (day) 19501 19502 19503 19504 195041; 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[O.0432]<(0.05) NS 0.0609 NS 0.199 NS 0.0648 21 NS NS NS <LLQ[O.o258]<(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 j.tg/mL). *Rat Number with "F" indicates fetal sample. NS: No sample. Page 13 of 24 PTI 0 99ADJA02.MLDOC 2 0 I I I .I .j 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[O.o483]<(0.05) NS 0.0664 0.0619 <LLQ[O.o449]<(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[O.o269]<(0.05) 0.102 NS NS 0.0833 0.219 - 'D Page 14 of 24 99ADJA02.MLDOC a, rn (D j ? i i 3 .i i i W 3 1 5 a m 0 Table 5: (Continued) Concentrationsof PFOS in Rat Serum Samples from Study #FACT-TOX-110Following the 0 mg/kg Dose Sampling Time (day) 19513 PFOS Concentration (pg/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 d L Q =Less than the Lower Limit of Quantitation (0.05 pghL). *Rat Number with "F" indicates fetal sample. NS: No sample. Page 15 of 24 99ADJA02.MI.DOC 2 4 1 I I L I W z Table 6: Concentrations of PFOS in Rat Serum Samples from Study #FACT-TOX-llO Following the 0.1 mgkg Dose ~ Sampling Time . (day) 19517 19518 ~ PFOS Concentration (pg/mL) Rat Number* 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 I *Rat Number with 'IF"indicates fetal sample. NS: Nosample. -0 Page 16 of 24 99ADJA02.MI.DOC a0) (D i i i i Table 6: (Continued) Concentrationsof PFOS in Rat Serum Samples from Study #FACT-TOX-110Following the 0.1 mgkg Dose NS 12.0 9.48 12.0 10.0 NS 11.6 NS 8.48 9.10 10.6 NS 9.62 11.2 8.78 7.80 NS 10.3 NS 7.65 7.57 10.1 NS 10.0 11.7 8.78 8.04 NS 7.31 NS 11.8 10.4 9.66 NS 6.19 10.8 3.04 10.4 NS NS NS I *Rat Number with "F" indicates fetal sample. NS: No sample. Page 17 of 24 99ADIA02.MI.DOC h) W 03 i i i i i W z Table 7: Concentrations of PFOS in Rat Serum Samples from Study #FACT-TOX-110 Followingthe 0.4 mg/kg Dose Sam~lingTime 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 Page 18 of 24 P-0 0 ;r 99ADJA02.MI.DOC w i '5. B 5ar Table 7: (Continued) Concentrationsof PFOS in Rat Serum Samples from Study #FACT-TOX-110 Followingthe 0.4 mg/kg Dose PFOS Concentration (pg/mL) A Sampling Time Rat Number* h) (day) 19542 19543 19543F 19544 19545 I9546 19546F 19547 19548 0 45.3 46.3 NS 45.8 47.9 43.5 NS 42.2 57.6 7 40.6 46.1 NS 42.8 NS 46.5 NS 42.2 60.0 15 44.0 43.9 NS 49.1 NS 47.3 NS 43.3 58.0 21 NS 23.0 34.1 NS NS 35.8 48.4 NS 65.7 P V 0 ?? 73 Page 19 of 24 99ADJA02.MLDOC nl 0 (D h) A P 0 0 u S ' 5a 0 nr 2 C Table 8: Concentrations of PFOS in Rat Serum Samples from Study #FACT-TOX-110 Following the 1.6 mg/kg Dose Q 5. Y 184 165 182 203 166 199 198 196 171 200 NS NS 160 189 195 186 NS 171 160 171 NS 160 NS NS 135 176 225 202 NS 164 192 215 NS NS NS NS NS 251 NS NS 237 79.4 NS: No sample. Page 20 of 24 P U 0 ;r 99ADJA02.MI.DOC Sampling Time (day) 19560F 0 NS 7 NS 15 NS 21 131 19561 193 181 219 NS PFOS Concentration (pg/mL) Rat Number* 19562 19562F 19563 185 NS 163 179 NS 168 142 NS 183 63.7 102 NS *Rat Number with "F"indicates fetal sample. NS: No sample. 19564 185 169 159 NS m A U 0 7 s Page 21 of 24 UmT) N A P 0 r0 i _i ? Table 9: Concentrations of PROS in Rat Serum Samples from Study #FACT-TOX-110Following the 3.2 mg/kg Dose u a 370 337 366 339 NS 334 NS 351 348 NS 357 NS 392 280 327 340 NS 323 NS 337 371 NS 305 NS 294 336 NS 314 NS 269 308 NS 317 NS 241 228 142 176 NS 165 166 123 166 I *Rat Number with "F"indicates fetal sample. NS: Nosample. Page 22 of 24 P U 0 ;r 99ADJA02.MI.DOC i Table 9: (Continued)Concentrationsof PFOS in Rat Serum Samplesfrom Study #FACT-TOX-110 Followingthe 3.2 mgkg Dose Sampling Time (day) 19573 19574 19575 PFOS Concentration(pg/mL) Rat Number* 19576 19577 19577F 19578 19579 19579F 19580 373 387 NS 378 *Rat Number with "F" indicates fetal sample. NS: No sample. w s ) 6 a a0 , B-0 30a) 3 .. ? u a r 2 a, a0 , Page 23 o f 24 99ADJAOZ.MI.DOC 10 P P Table 10: Repeat Analysis of Rat Serum Samples from Study #FACT-TOX-110 Rat Treatment Time original Number Conc. &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-407.59183400) 19579 5 Day 0 >ULQ-403.4253>(400) XJLQ: Greater than the Upper Limit of Quantitation. REASONS FOR REASSAY: 1). Run 6 concentration exceeded calibration range. Original Curve Number 6 6 6 6 6 6 6 Reason for Reassay 1 1 1 1 1 1 1 Reassay Conc. pg/ rnl 392 345 366 393 413 417 387 REASONS FOR REPORTEE CONC: 1). Run 7 concentration within calibration range. Reassay Curve Number 7 7 7 7 7 7 7 Reported Conc. pg/d 392 345 366 393 413 41 7 387 Reason for Reported Conc. 1 1 1 1 1 1 1 Page 24 of 24 99ADJA02.Ml.DOC 3lkl Medical DepartmentStudy: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 , ADVANCED BIOANALYTICAL SERVICES, INC ADDENDUM TO THE BIOANALYTICAL REPORT r, Title: - Addendum QuantitativeDeterminationof Perfluorooctanesulforiate(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 Authors: David J. Anderson, M.S. Amie J. Prince, B.S. Holly D. Ross, M.S. Prepared For: 3M EnvironmentalTe,chnology and Safety Services St. Paul, MN 55133-3331 Number of Pages: 3 m\ 15 Catherwood Road m Ithaca. New York 14850 m (607)266-0665 m Fax (607)266-0749 3M IInvironmental Laboratory Page 246 3iW Cedical Department Study: T-6295.12 .4 I ,. Analytical Report: FACT TOX-110 LRN-U2849 ADVANCED BIOANALITICAL SERVICES Title: SIGNATURE PAGE Addendum - QuantitativeDetermination of Perfluorooctanesulfonate (PFOS) in Rat Serum from Study #FACT-TOX-110Using Turbo Ion Spray LC/MS A B S Report No.: 99 Reported by: Research Sciktist 13 m 4 9 Date Reviewed by: Authorized for Release by: Date Associate Auditor +- A John R. erkins, Ph.D. 13. a u . 5 5 Date Assistdt ScientificDirector Accepted by: 3M Study Director 64 /f UP+ 419 Date 3M Environmental Laboratory ADVANCED BIOANALYTICAL SERVICES, INC. Page 2 of 3 99ADJA02.MLADD.DOC Page 247 3M Medical DepartmentStudy: T-6295.12 Ah I 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. LC 31vl EnvironmentalLaboratory ADVANCED BIOANALYTICAL SERVICES, INC. Page 3 of 3 99ADJA02.MI.ADD.DOC Page 248 3NI Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 RE-ISSUEDANALYTICAL REPORT STUDY TITIZ Oral (Gavage) Pharmacokinetic Study of PFOS in Rats DATA REOUIREIvlENTS Analytical Method Requirements STUDY DIRECTOR Marvin T. Case, 3M PRINCIPAL INVESlrIGATOR Enaksha Wickremesinhe, Centre RE-ISSUED ANALYTICAL REP0R.T COMPLETIONDATE October 19,2000 PERFORMING LABORATORY 1' TESTING FACILITY Centre Analytical Laboratories,Inc. (Centre) 3048 Research Drive State CoIlege, PA 16801 Phone: 814-231-8032 STUDY S P O N i m 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 3NI EnvironmentalLaboratory Page 249 3Nl Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 SponsorProtocol 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 Toxicdogy 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 (analytical standards) used in this study (PFOS ' and THPFOS) have not been characterizedunder 21 CFR 58.105 2. The automated data collection systems used in this study are not fully compliant with 21 CFR 58.130(e). Enaksha Wickremesinhe,Ph.D* - Date Principal Investigator Centre Analytical Laboratories,Inc. *No longer employed at Centre. However, facility management, Rick Grazzini will be signing in the principal investigator'sstead. Marvin T Case, Ph.D. Study Director 3M Toxicology Services - a30& Amd Date Centre Analytical Laboratories, Inc. 3NI Environmental Laboratory Page 2 of 103 Page 250 3NI Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 OUALITY 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 JnsDected 51 17/00 Date Reported to Centre Management 6/1/00 2. Extraction 6/27/00 711 1/00 3. Raw Data Review 7/10,13,14,20/00 7/29/00 4. Draft Report Review 8/3-4/00 81 15/00 5. Final Report Review 9/8/00 911 1/00 Date Reported to Study Director and ponsor Management 7/14/00 7/14/00 8/25/00 8/25/00 911 1/00 u Naomi Lovallo Quality Assurance Auditor . - Centre Analytical Laboratories, Inc. 3NI 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 AlJTHENTICITY 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: Enaksha Wickremesinhe, Ph.D.* Date Groupmeam Leader Centre Analytical Laboratories, Inc. *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: ohn Flaherty / Date L b o r a t o r y Manager Centre Analytical Laboratories, Inc. Study Director, 3 M Marvin T. Case, Ph.D. 3M Toxicology Services - 9-3u ! 2ruc, Date Centre Analytical Laboratories, Inc. 3rd EnvironmentalLaboratory 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 SponsorProtocol No:FACT-TOX-110 STUDY IDENTIFICATION Oral (Gavage) PharmacokineticStudy of PFOS in Rats SPONSOR PROTOCOLNUMBER:FACT-TOX-110 TYPE OF STUDY Oral (Gavage) Pharmacokinetic TEST SYSTEM: Rat Feces SPONSOR: STUDY DIRECTOR: Perfluorooctmesulfonic acid potassium salt (T-6295) 3M Toxicology Services - Medical Department 3M Center, Eluilding 220-3502 St. Paul, MN 55144-1000 From 6/10/99 to 2/10/00: Kris Hansen, PhD. 3M EnVirOMlentd Technology and Safety Services Phone: (651) 778-6018 From 2/10/00 to Present: Marvin T. Case, D.V.M., Ph.D. 334 ToxicologyServices Phone: (651) '733-5180 TESTING FACILITY. Centre Analytical Laboratories,hc.. 3048 Research Drive State College, PA 16801 PRINCIPAL INVESTIGATOR Enaksha Wickremesinhe, Ph.D. Cenm Analytical Laboratories, Inc. Phone: (814) 2!31-5032 AVALYTICAL PHASE mmmm: Study Initiation Date: 06/10/99 Analytical S t a t Date: 06/09/00 ' k d y t i c d Termination Date: 07/01/00 Centre Analytical Laboratories, Inc. 3M EnvironmentalLaboratory Page 5 of 103 Page 253 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2.849..-.. Centre Study No.: 023-016 Sponsor Protccd No: FACT-TOX-I10 PRO.JECT PERSONNEL The Study Director,for this project was Marvin 7'. Case at 3M Toxicology Services and the Principal Investigator for this project at Centre Analytical Laboratories, Inc. was Enaksha Wickremesinhe. The following pemnn!:l from Centre Analytical Laboratories, Inc., were associated with various anillytical phaseir of the study: &DE Enaksha Wickremesinhe Emily Stauffer Karen Smith Tiffany Proctor Rickey KelIer Lawrence Ord - - Title CirouplTeam Leader Scientist Scientist Tlechnician Simple Custodian Sample Custodian Centre AnaIyticil Laboratories. Inc. 3NI Environmental Laboratory Piqe 6 of 103 Page 254 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 OF CONTENTS Pa..e T'lTLE PAGE ....................................................................................................................... 1 GOODLABORATORYPRACTICEC O M P ~ W SETATEMEhT............................. 2 QUAIJ"Y ASSTJRANCE STA- ..........................................................................3 CERmCAxON OF A m C m...................................... ;.................................... 4 STUDY IDENTIFICATION............................................................................................... 5 . PROJEn pERs0NiVE.L.................................................................................................... 6 TABLE OF CO"S .................................................................................................... 7 LlSTOFTABES .............................................................................................................. 8 LIST OF R G W .............................................................................................................. 9 UST OF A p p m I a .................................................................................................... 10 1.0 suMMpA.Ry ................................................................................................................ 11 2.0 O B J E m............................................................................................................... 11 3.0 lj"RODUrnON ........................ .............................................................................. 11 4.0 TEST SYSTEM ......................................................................................................... I1 5.0 REFERENCEMATERlAL....................................................................................... 12 6.0 ExpERlMENTALDESIGN...................................................................................... 13 7.0 DESCRIPTIONOF A N A L ~McE"I~3011....................................................... 13 7.1 Extraction Procedure ........................................................................................... 13 7.2 Preparation of Standards and Eom'ficationSolutions.......................................... 13 7.3 Chromatography ................................................................................................... 14 7.4 InstrumentSensitivity........................................................................................... 14 7.5Descriptionoflnstrumentand OperatingConditions.......................................... 14 7.6 QumtitationandExampleCJculation................................................................ 16 8.0 RESULTS DISCUSSION.............................................................................. 18 9.0 CIRCMsTmCJ3 THATMAY HAVE A I ; F E C m THE DATA.................... 18 10.0 -ON OFDATA AM> SAMPLES............................................................. 18 11.0 TABLES ................................................................................................................... 19 12.0 R G m S ................................................................................................................. 40 13.0 A~PWICES .......................................................................................................... 48 Centre Analytical Laboratories. Inc. 3NI EnvironmentalLaboratory Page 7 of 103 Page 255 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 TABLES Page Table I. Summary of PFOS residues in Matrix Blanks and Matrix Zero Blanks.............20 Table II. Summary of PFOS residues in GI-Control-GDO.............................................. 22 Table ID. Summaryof PFOS residues in G2-0.1MD on GDO ...................................... 23 Table IV. Summvy of PFOS residues in G3-0.4lW on GDO...................................... 24 Table V. Summary of PFOS residues in G4-1.6MXD on GDO ....................................... 25 Table VI. Summaryof PFOS residuesin G5-3.234KD on GDO...................................... 26 Table VII. Summaryof PFOS residues in G1-Control on GD7....................................... 27 Table wt Summary of PFOS residues in G2-O.lMKD on GD7 ................................... 28 Table iX. SummG of PFOS residuesin G3-0.4hED on GD7...................................... 29 Table X. Summary of PFOS residues in G4-1.6MICDon GD7 ....................................... 30 Table XI. Summary of PFOS residues in G5-3.2MJXD on GD7...................................... 31 Table XIL S u m w of PFOS residues in G1-Control Dn GD15..................................... 32 Table W L Summary of PFOS residues in G2-0.1MxD on GD15 ................................. 33 Table XIV . Summary of PFOS residues in G3-0.4MXD on GD15................................. 34 Table XV . Summary of PFOS residues in G4-1.6EdKD on GD15 .................................. 35 Table XVI. Summary of PFOS residues in G5-3.2MKD on GD15................................. 36 Table XVIf. Summary of PFOS residuesin Gl-Control on GD21.................................. 37 Table XWI. Summaryof PFOS residues in G2-0.lMKD on GD21 .............................. 37 Table XM. Summary of PFOS residues in G3-0.41MKD on GD21................................. 38 Table XX S u m q of PFOS residues in G4-1.6MKD on GD21.................................. 38 Table lw. Summaryof PFOS residues in G5-3.21vlK.D on GDZ1................................. 38 Table XXII. Summary of PFOS recoveries in FortifiedSamples.................................... 39 Centre AnalyticalLaboratories. Inc. 3NI EnvironmentalLaboratory ' P?ge 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 Protocoi No: FACT-TOX-110 Figure 1. Figure 2. - LIST OF FIGIJRES Page Typical Calibration Curve for PFOS.............................................................. 41 TypicalMean Response Factor for THIyFOS...........................,....................42 Figure 3. C&matogram Representing B 5 n&d. extracted standard for PFOS and 250 n g / d fortificationof THPFOS..................................................................... 43 Figure 4. ChromatogramRepresentins a 125n@/mt extracted standard for PFOS and . 250 n g / d fortification of THPFOS..............,................................................ 44. Figure 5. Chromato- Representing Control Rat Feces for PFOS and THPFOS (CentreID:0004277Blank B,Set: 062300A).............................................. 45 3 Figure 6. Chromatogram Representing Control Rat Feces Fortified with 50 ng ofPFOS and 500ng of THPFOS (Centre ID: 0003981 Spk A,Set: 061400A).........46 Figure 7. Chromatagramof Rat Feces Sample from G2 on GDO (Centre ID: 0003994, Set: 061400A) ............................................................................................... 47 Centre Analytical Laboratories.hc. 3NI Environmental Laboratory Page 9 of 103 Page 257 3Ml Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U284.9.. . CentreStudy No.:023-016 Sponsor Protocol No: FACT-TOX-110 LIST OF APPENDICES AppendL. A Study ProtocolFACT-TOX-110(Centre Study No. 023-016) and Amendments and Deviations ...........-............,...................................... - Page ..... 49 Appendix B Determination of FIuorochemicalResidues in Monkeymat Feces by LC/MS/MS (Revision 2) Method +OOM-023-003r,evision 2 and Deviations and iviodifications .............................................................76 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 10 of 103 Page 258 3kl Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023416 Sponsor Protocol No: FACT-TOX-I10 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 Monkeymat Feces by LC/MS/MS, revision 2" (Centre method number: OOM-023403,revision 2). The limit of quantificationfor PFOS in rat feces was 10ppb. Residues (corrected for purity) ranging from non-detected levels to 59386.8 ppb were found in the rat feces samples. Fortification recoveries ranged from 71-12896 with an average of 98% and relative standard deviation of 15%. 2.0 0BJECTI:VE 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 FluorochemicalResidues in Monkeymat Feces.byI-CIMSIMS,revision 2." 3.0 INTRODUCTION This report details the results of the residues of E'FOS detected in rat feces, using the analytical method entitled, "Determination of FIuorochemicaI Residues in Monkeymat 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 !>,2000, and the analytical termination date was July 1,2000. 4.0 TEST SYS'IW 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 bl.anks was purchased from Lampire Biological Laboratories, Inc., Pipersville, PA and received at Centre frozen on dry ice on Centre Analytical Laboratories, Inc. 3NI EnvironmentalLaboratory Page`11of 103 Page 259 3NI Medical Department Study: T-6295.12 I Analytical Report: FACT TOX-110 LRN-U2849' Centre Study No.: 023-016 Sponsor Protocol No: FAm-TOX-110 May 4, 2000 and June 22, 2000, logged in by Centre personnel and placed in frozen storage (<-lOC). 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 MkTERIAL The analytical standard PFOS (as the potassium salt) was received at Centre on November 12, 1999 and the surrogate standard TIBFOS 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. ComDound PFOS THPFOS Centre Control No. 99-023-002 99-023-011 Batch No. 217 53406 %tv (%I Expiration Date 86.3 0813 1/01 NA 01/01/10 Molecular structuresof PFOS and THPFOS are given below. PFOS . Chemical Name: Peffluorooctane sulfonate Molecular weight: 499 (CsFd03-) 0 Note: The neutral molecule and standard foim from which PFOS (anion)is derived, is potassium perfluorooctane sulfonate [C&7S03K], molecular weight 538. THPFOS Chemical Name: 4-H, peffluorooctane SulfOiniC acid MolecuIar weight: 428. 1-H,1-H,2-H,2-H, CsF&OjH Centre Analytical Laboratories. hc. 3NI Environmental Laboratory Page 12 of 103 Page 260 3Nl Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 SponsorProtocol No: FACT-TOX-I10 6.0 EXPERIMENTALDESIGN 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 12 I6 samples. The extncts were analyzed by Lc/Ms/Ms. Samples with residues outside the linear range of the calibration curve were diluted and re-analyzed. 7.0 D E W T I O N OFANA:LM?CAL METHOD Analytical method entitled "Deteimination Of Fluorochemical Residues in Monkeymat Feces by Lc/MS/MS (Revision 2)" was used for this study. 7.1 Extraction Procedure One gnm 1:0.05 of sample was weighed into 20 polypropylenescintillation vids and fortified to the vi (if d, necessary> using disposable micropipettes. capped tightly, and placed on a wrist-action - Ten mL of acetonitrile was added shaker for 30 min. The samples were filtered through a glass acrodisc filte:. The filtered extract was passed through a conditioned c h o n SPE column and collected. Tlie columns were then eluted with -10 mL acetonitrile followed by -20 mL 9O:lO acetonitde:2% ascorbic acid in methanol. The combined extracts were evaporated down to dmost dryness with a rotary evaporator and then reconstituted with methanol, making find volume 2 mL. The,samples were analyzed using electrospray Lc/Ms/Ms. 7.2 Preparation of Shndards and Fortification S O ~ U ~ ~ O ~ S Stmdard solutions were prepared on May 3, 2000 as specified in Centre Analytical Laboratories' analytical method entitled, "Determiliation of Fluorochemical Residues in Monkeymat Feces by LC/MS/MS (Revision 21." .4n individual stock standard soIution of PFOS was prepared ;It a concentration of 100 p g h L by dissolving 10 mg of the standard (corrected for salt punty 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&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 p$mL solution of THpFOS w s prepared in the same fashion described above. A 2.5 pg/mL fortification standard of PFOS was i~repmdby taking 25 mL of the 10 pglmL 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 p g h L PFOS standard was prepared by taking 20 mL of the 2.5 pg/mL PFOS Centre Analytical hboriitories, Inc. 3N1 Environmental Laboratory Page 13of 103 .. ! i ! 1 \ i f i ! i ., Page 261' 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 standard and bringing to 100 mL with methanol. To make the 0.1 pgJmL PFOS fortification standard, 20 mL of the 0.5 P g h L of PFOS was brought to 100 d with methanol. Calibration standards were processed through tl.,e extraction,proctdure, identical to the samples. The fortification of the standards before extraction was done according to the following table: . Conc. of PFOS Fortification Fortification Volume (pL) Solution (Pg/d) Weight of Control h n p k (g) k 0.05 0.1 100 . 1.0 0.1 200 1 .o 0.5 100 1.0 0.5 200 1.0 2.5 100 1.0 2.5 200 1 .o * 2.5 pg/d THPFOS fortification solution. ,Fort.Level of Extracted Calibration Standard (PDb) 10 20 50 100 250 500 VOI. of Surrogate Standard* added (pL) 200 200 200 200 2--00- 200 The stock standard solution and all fortification and calibration standard solutions were - stored in a refrigerator (4" f 2C) when not in use. Documentation of standard preparation and extr;lction can be found in the raw data associated with this report. 7.3 Chromatography 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 Mn. and - 5.0 min., respcctivcly, with no significant interfering peaks in the control matrices corresponding to either of the andyte retention times. 7.4 Instrument Sensitivity The smallest standard mount injected during the chromatographicrun was equivalent to 5 ng/m.L of PFOS md 250 ngmL of THPFOS in the rat feces matrix. 7.5 Description of Instrument and Operating Conditions A Micromass Quattro Ultima LC/MShfS coupled to a Hewlet Packard HPLC system was used. Data acquisition and processing were performedusing Masslynx 3.4 software. Detailed operating conditions are listed below: Centre Analytical Laboratories, Inc. Page 14 of 103 . 3NI EnvironmentalLaboratory Page 262 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 , Centre Study No.: 023416 Sponsor Protocol No: FACT-TOX-I 10 Instrument: Micromass Quattro Ultima ELECTROSPRAY ION SOURCE; Capillary: 3.0 kV Hexnpole 2: 0.3 V Hexapole 1: 0.1 V Source Block Temp.: 1 0 0 0 ~ A p m 1: 0.2 V Desolvation Temp.: 350C ANALYZER: LMRes 1: 10.5 V HM Res 1: 10.5V n e r g y 1: 1.0 V Entrance: -2 V Exit: 2 V LAMRes 2: 13.0V MRes 2: 13.0 V Energy 2: 2.0 V hlultiplier: 650 V GAS FLOWS AM)PRESSURl$: Desolvation N2 Flow Rate: -650 L/hr . Nebuliser N2Flow Rate: -150 L h Gas Cell Pressure: -0.003 mliar Computer: COMPAQProfessional Workstation m o o Software: MicrosoftWindowsNT: Version4 BuiId 1381: ServicePack 5 M ~ C ~ O WLiSmited: AhSSlyrUi3.4 Build 004 HPLC Equipment: HewIettPachrd (HP) Series 1100 HPBinaryPump -' HP Vacuum Degasser HP Autosampler HP ColumnOven HPLC Column: Column Temperature: Mobile Phase (A) : Mobile Phase (B) : Genesis C-8, 5 cm x 2.1 mm i.ci. x 4 1.1 35C ' 2 mM Ammonium Acetate in Type I Water Methanol Gradient: Injected Volume: .Ions monitored : _AnJvte PFOS THPFOS Time (min) 0.0 0.4 1.0 1.0 1.5 10.5 11.0 145 15.0 10 jlL %A 60.0 60.0 10.0 10.0 0.0 0.0 60.0 60.0 60.0 Transition Monitored 499. -) 9.9. 427+80 Dwell (secsl &I1 . 0.7 ' 0.2 %B 40.0 40.0 90.0 90.0 100.0 100.0 40.0 4Q.O 40.0 Flow Rate (mumin) 0.3 0.3 0.3 0.3 0.3 n-.d, 0.4 0.4 0.3 Enerev lev) Cone 4-7 7I "L Xi 34 Centre Analytical Laboratories, Inc. 3NI Environmental Laboratory Page 15 of 103 Page 263 3M Medical DepartmentStudy:T-6295.12 Analytical Report: FACT TOX-110 ' LRN42849 Centre Study No.: 023-016 , SponsorProtocol No: FACT-TOX-110 7.6 Quantitation and Example Calculation 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 l/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. Eauation 1: Analyte found (ng/mL) = Peak m a - interceut) slope Eauation 2 Analyte found (ppb) =janalvte'found h.hnLl x final vol. (mLl x DF) stamplewt. (g) where DF = dilution factor. For samples fortified with known amounts of analytes prior to extraction, use Equation 3 to calculate the percent recovery. Eauation 3: Recovery (9%) = [analytefound (ng/mL)x final vol. (.inL)xDF] xloo analyte added (ng) Eauation 4 This calculationwas only performed on actual s:mples and not QC recoveries. Corrected analyte found (ppb) = analyte found (pptj) X 9% punty 9% punty for PFOS lot 217 = 86.9%(0.869) An example of a calculation using m 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 = 32320 intercept slope dilution factor = 2653.87 = - - 595.935 1 Centre Analytical Laboratories, Inc. Page '16of 103 3NI Environmental Laboratory Page 264 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849. ng added (fort level) = final vol. = sample wt. = 100 2mL 1.01 1; Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 From equation 1: Analyte found (ng/mL) = ~32320-2653.871 595.935 = 49.8 ng/mL From equation 2: Analyte found (ppb) = 149.8 X 2 mL X 11 1.01 e; = 98.6 ppb From equation 3: % Recovery = (49.8 n d d x 2 1nLx 1) x 100 100 ng = 100% Note: This example calculation was done using rounded numbers, and therefore may be slightly differentfrom the values shown in the RAW DATA. The amount of surrogatestandardTHPFOS found was calculated using the following equation: Analyte found (nglmL): peak area mean response factor I Other statisticalmethods used in analyzing this data..were: Standard Deviation= n Relative Standard Deviation (RSD or Coeficient of Variation(CV))= Standard Deviation x 100% .. Mean Centre AnaIytical Laboratories,Inc. 3NI EnvironmentalLaboratory 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 leveis. 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 inteiral are detailed in Tables XI-=. For this report the residues found in Tables I-)MI 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 s ~ m m oqf 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 11-7. 9.0 CIRCUMSTANCES THATMAY HAVEAFFECTED THEDATA Electronic records are not fully compliant withi 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 RETIENTION OF DATXAND SAMPLES W h e n the final report is complete, dl 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 substancesare achived by the sponsor. Centre Analytical Laboratories, Inc. 3NI EnvironmentalLaboratory Page IS of 103 Page 266 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.:023416 Sponsor Protocol No:FACT-TOX-I 10 11.0 - TA:BLES, Centre Analytical Laboratories, Inc. 3Nl Environmental Laboratory Page 19 of 103 Page 267 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 Table 1. Summary of PFOS residues in :MatrixBIanks and Matrix Zero Blanks ND = Not Detected and NQ = Not Quantifiable Sponsor Centre Residue 0 . - u c * e d Set Date Found Residue Found ID ID Number Extracted (ng/g) (ndb9 na 0004277 Blank A 061200AR 6/12/00 ' ND ND na 0004277 Blank B 061200AR 6/12/00 ND ND na 0004277 Zero Blank C 06 1200AR 6/12/00 ND ND na 0004277 Zero Blank D 061200AR 6/12/00 ND ND na 0004277 Blank A 0614OOA 6/14/00 NQ NQ na 0004277 Blank B 061400A 6/14/00 NQ NQ na 0004277Zero Blank C 0614OOA 6/14/00 NQ NQ na 0004277Zero Blank D 061400A 6/14/00 ND ND na 0004277 Blank A 061500AD 6/15/00 NQ NQ na 0004277Blank B 0615OOAD 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 0616OOA 6/16/00 2.0 1.7 . na 0004277 Blank F 061600A 6/16/00 NQ NQ na 0004277Zero Blank G - 061600A 6116/00 NQ NQ na 0004277 Zero BlankH 0616OOA 6/16l00 na 0004277 Blank A 0619ooAD 6/19/00 na 0004277Blank B 0619ooAD 6/19/00 na 0004277 Zero Blank C 061900AD 6/19/00 na 0004277Zero Blank D 0619ooAD 6/19/00 na ooO3466 Blank A 062000A 6/2o/oo na 0003466 Blank B 062000A 6/20/00 na 0003466 Zero Blank C 062000A 6IZWOO na 0003466ZeK, Blank D 062000A 6/20/00 na 0004277 Blank A 062100A 6/21/00 na 0004277 Blank B 062100A 6/21/00 na 0004277 Zero Blank C 062100A 6/21/00 na 0004277 Zero Blank D 062100A 6/21/00 na 0004277 Blank A 062200A 6/22/00 na 0004277 Blank B 062200A 6/22/00 na 0004277 Zero Blank C 062200A 6/22/00 na 0004277 Zero Blank D 062200A 6/22/00 na 0004277 Blank E 062200B 6/22/00 na 0004277 Blank F 062200B 6/22/00 na 0004277 Zero Blank G 0622OOB 6/22/00 Centre Analytical Laboratories, Inc. Page 20 of 103 3M Environmental Laboratory Page 268 3NI Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 fabie I (cont.) Summary of PFOS residues in Matrix Blanks and latrix Zero Blanks ND = Not Detected and NQ = Not Quantifiable Residue Corrected Residue Sponsor Centre ID Set Date Found Found ID Number ktracted (np/,g) na 0004277 Zero Blank H 062200B 6/22/00 ND bg/e) 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 062300A 6/23/00 ND ND :la 0004277 Blank A 062300B 6/23/00 NQ NQ la 0004277 Blank B 062300B 6/23/00 NQ NQ la 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 P 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 0005738Blank E 0626M1B 6/26/00 ND ND na 0005738 Blank F 062600B 6/26/00 ND ND nil 0005738 Zero Blank G 06260B il26400 ND .ND na 0005738Zero 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 na 0005738 Zero Blank D 062700A m7/00 ND ND ,m na 0005738 Blank A 062800A 6/28/00 ND ND na 0005738 Blank B 062800A 6/28/00 ND ND na 0005738ZeroBlaak C 062800A 6/28/00 ND ND na 0005738 ZeroBlank D 062800A 6/28/00 ND ND na 0005738 Blank A 062900A 6/29/00 30.1 26.2 na 0005738 BlankB 062900A 6/29/00 ND ND na 0005738 Zero Blank C 062900A 6/29/00 m ND na 0005738 Zero Blank D 0629OOA 6/29/00 ND ND ria 0005738 Blank A 063000A 6/30/00 ND ND na 0005738 Blank B 063000A 6/30/00 m ND na 0005738 ZeroBlank C 063000A 6/30/00 ND ND na 0005738 Zero Blank D 063000A 6/30/00 ND ND AVERAGE: 0.8 0.7 STANDARD DEVIATION 4 3 Centre Analytical Laboratories, Inc. Page 21 of 103 3NI EnvironmentalLaboratory Page 269 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 Table II, Summary of PFOS residues in GI-Control-GDO Sponsor CentreID Set Corrected Date Residue Found ResidueFound ID Nurnber Extracted 1950lF,FO,G1-Control-GDO 0003977 061200AR 6/12/00 19502F,FO,G1-Control-GDO 0003978 06l200AR 6/12/00 l95O3F,FO,Gl-Control-GDO 0003979 061200AR 6/12/00 19504F,FO,G1-Control-GDO 0003980 061200AR 6/12/00 19505F.FO.Cl-Conml-GDO 0003981 061200AR 6/12/00 19506F.FO.Gl -Control-GDO 0003982 061200AR 6/12/00 19507F.FO.G 1-ControI-GDO 0003983 061200AR 6112/00 19508FP0,G1-Control-GDO Ow3984 061200AR 6/12/00 19509F~0,Gl-Control-GDO 0003985 061200AR 6/12/00 195lOF,FO,Gl-ControI-GDO 0003986 061200AR 6/12/00 1951lF,FO,Gl-Control-GDO 0003987 061200AR 6/12/00 19512FPO.G1-Control-GDO 0003988 061200AR 6/12/00 19513F,FO,Gl-ControLGIXY 0003989 061200AR 6/12/00 19514F,FO,Gl-Control-GDO 0003990 061200AR 6/12/00 19515F,FO,Gl-ControlGDO 0003991 061200AR 6/12/00 195MF,FO,Gl-Control-GDO 0003992 061200AR 6/12/00 AVERAGE: STANDARD DIEVIA'MON (n&) 0.0 0.0 5.9 0.0 0.0 30.1 0.0 5.9 40.0 0.0 0.0 0.0 0.0 0.0 0.0 -60.9 . 8.9 18 (ngld 0.0 0.0 5.1 0.0 0.0 26.2 0.0 5.1 34.8 0.0 0.0 0.0 0.0 0.0 0.0 52.9 7.8 16 Centre Analytical Laboratories. Xnc. 3Wl Environmental Laboratory Page 22 of 103 Page 270 3NI Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 CentreStudy No.:023-016 SponsorProtocol No:FACT-TOX-110 Table III. Summary of PFOS residues in G2-0.1Mfa) on GDO Sponsor CentreID Set Corrected Date Residue Found Residue Found ID Number Extracted 19517F,FO,G2-O.lMKD-GDO0003993 061400AD 6/14/00 (ndg) 700.8 fndg) ~ 609.0 195 18F,FO,G2-0.1MKD-GDO 0003994 195 19F,FO,G2-0.1MKD-GDO 0003995 1952OF,FO,G2-0.IMKD-GDO 0003996 19521F,FO,G2-0I.MKD-GDO 0003997 061400A 061400A 06 14OOAD 06 14OOA 6/14/00 6/14/00 6/14/00 6/14/00 455.7 389.7 675.5 494.8 396.0 338.6 587.0 430.0 19522F,FO,G2-0I.MKD-GDO 000399a 061400A 6/14/00 381.0 331.1 19523F,FO,G2-0.1MKD-GDO 0003999 061400A 6/14/00 458.2 398.2 19524F,FO,G2-0.IMKD-GDO 0004Ooo 0614OOAD 19525FZO,G2-0.IMKD-GDO OOO4001 0614OOA 19526F,FO,G2-0.IMKD-GDO 0004002 0614OOA 6/14/00 6/14/00 6/14/00 832.8 473.0 446.1 723.7 411.0 387.7 19527F,FO,G2-0.1MKD-GDO 0004003 061400A 6/14/00 497.4 432.2 19528F~O,G2-0.lMKPGDo0004004 061400AD 6/14/00 700.7 608.9 19529F,FO,G2-0I.MKD-GDO 0004005 061400AD 6/14/00 796.9 692.5 19530F,FO,G2-0.IMKD-GDO 0004006 061400A 6/14/00 393.7 342.1 1953 lF,FO,G2-0.lMKD-GDO 0004007 0614ooAD 6/14/00 734.7 -638.5 19532F,FO,G2-0.1MKD-GDO 0004008 061400AD 6/14/00 748.2 650.2 AVERAGE: STANDARD DIEVIATION 573.7 160 498.5 139 Centre Analytical Laboratories,Inc. 3NI EnvironmentalLaboratory 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 SponsorProtocol No:FACT-TOX-I10 Table IV, Summary of PFOS residues in.G3-0.4MKD on GDO Sponsor CentreID Set Date Residue Found ID Number Extracted 19533F,FO,G3-0.4MKD-GDO 0004009 061500A.E 6/15/00 (ne&) 2598.1 19534F,FO,G3-0.4MKD-GDO 0004010 061500AD 6/15/00 3750.7 19535F,FO,G3-0.4MKD-GDO 0004011 061500AD 6/15/00 2801.3 19536F,FO,G3-0.4h4KD-GDO0004012 061500AD 6/15/00 1953TF,FO,G3-0.4MKD-GDO 0004013 061500AD 6/15/00 3174.3 2777.9 19538F,FO,G3-0.4MKD-GM) OOO40 14 061500AD 6/15/00 3083.8 19539F,FO,G3-0.4MKD-GDO 0004015 061500AD 6/15/00 3680.4 19540F,FO,G3-0.4rWa)-GDO0004016 061500AD 6/15/00. 2044.0 19541F,FO,G3-0.4MKD-GDO 0004017 061500AD 6/15/00 1~~~~F,FO.G~-O.~M0K00D40-1G8 DO061500AD 6/15/00 2104.0 3290.2 19513F,FO,G3-0.4MKD-GDO OOO4019 061500AD 6/15/00 2113.4 19544F,FO.G3-0.4MKD-GDO 0004020 061500A.D 6/15/00 3430.8 19545F,FO,G3-0.4MKD-GDO 0004021 061600AD 6/16/00 2755.3 19546F,FO,G3-0.4MXD-GDO 0004022 061500AD 6/15/00 2334.2 19547F,FO,G3-0.4MKD-GDO OOO4023 061500AD 6/15/00 2215.6 19548F,FO.G3-0.4MKD-GDO 0004024 061500AD 6/15/00 2334.8 AVERAGE: 2780.6 STANDARD DIEVIATION 570 Corrected Residue Found (@?z) . 2257.7 3259.4 2434.3 2758.5 2414.0 2679.8 3198.3 1776.2 1828.4 2859.2 1836.5 2981.4 2394.4 2028.4 1925.4 2028.9 24163 496 Centre Analytical Labontories. Inc. 3NI Environmental Laboratory Page 24 of 103 Page 272 3NI Medical DepartmentStudy: T-6295.12 i Analytical Report: FACT TOX-110 LRN-U2849 CentreStudy No.:023-016 SponsorProtocol No:FACT-TOX-110 Table V. Summary of PFOS residues in G 4 - 1 . m on GDO -Sponsor ~ ~~ - Centre ID Set Number Date Residue Found Extracted (nglg) 19549F,FO,G4-1.6MKD-GDO W O 2 5 061600AI) 6/16/00 14122.8 1955OF,FO.G4-1.6MKD-GDO 0004026 061600AI) 6/16/00 16858.7 1955IF3F0,G4-1.6MKD-GD00004027 061600AIk 6/16/00 10394.9 19552F,FO,G4-1.6MKD-GW 0004028 061600AI) 6/16/00 13143.8 19553F,FO,G4-1.6MKD-GDO 0004029 061600AT) 6/16/00 13712.0 19554F,FO,G4-1.6MKDGW 0004030 061600M~ 6/16/00 15507.9 19555F,FO,G4-1.6Mfll)-GDO 0004031 061600AT~ 6/16/00 10328.8 19556F,FO~G4-1.6MKD-GDO0004032 0616OOAU 6/16/00 10253.1 19557F,FO,G4-1.6MKD-GDO 0004033 061600A.D 6/16/00 8961.3 19558F,FO,G4-1.6MKD-GDO 0004034 061600A1c1 6/16/00 6845.2 19559F,FO,G4-1.6MKD-GDO0004035 061600AEi 6/16/00 7677.3 1956OF,FO,G4-1.6MKD-GDO 0004036 061600AcI 6/16/00 16518.0 1956lF,FO,G4-1.6MKD-GDO 0004037 06160OAE~ 6/16/00 12121.6 19562F,FO,G4-1.6MKD-GDO 0004038 0616OOAC~ 6/16/00 9585.4 19563F3F0,G41-.6MKD-GW 0004039 061600AD 6/16/00 7465.7 , 19564F,FO,G4-1.6MKD-GM) o004040 061600AD1 6/16/00 16957.0 AVERAGE: 119033 STANDARDDEVIATION: 3453 Corrected Residue Found (ng/g) 12272.7 14650.2 9033.2 11422.0 11915.7 13476.4 8975.7 ' 8909.9 7787.4 5948.5 6671.6 14354.1 10533.7 8329.7 6487.7 14735.6 10344.0 3000 Centre Analytical Laboratories, Inc. 3Mi EnvironmentalLaboratory Page25 of 103 Page 273 3b1 Medical Department Study: T-6295.12 Analytical Report:FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 SponsorProtocol No:FACT-TOX-110 Table VI. Summary of PFOS residues in G5-3.2MKD on GDO Sponsor CentreID Set Corrected Date Residue Found Residue Found ID Number Extracted 19565F.FO,G5-3.2MKD-GDO 0004041 061900AD 6/19/00 (ndc) 23450.7 (iidJ) 20378.7 19566F,FO,G5-3.2MXD-GDO 0004042 061900AD 6/19/00 29548.0 25677.2 19567F,FO,G5-3.2MKD-GW 0004043 061900AD 6/19/00 303 10.7 26340.0 19568F,FO,G5-3.;IMKDGDO (3o04044 0619oOAD 6/19/00 24160.4 20995.4 19569F,FO.G5-3.2MKD-GDO 0004045 061900AD 6/19/00 27052.1 23508.3 1957OF,FO,G5-3.2MKD-GDO 0004046 061900AD 6/19/00 19671.6 17094.6 19571F,FO,G5-3.2MKD-GDO 0004047 061900AD 6/19/00 29872.2 25958.9 19572F,FO,G5-3.2hfKD-GDO 0004048 0619oOAD 6/19/00 25395.0 22068.3 . 19573F,FO,G5-3.2MKD-GDO 0004049 061900AD 6/19/00 23006.0 19992.2 19574F,FO,G5-3.2MKD-GDO 0004050 06190&4D ' 6/19/00 29300.3 25462.0 19575F,FO.G5-3.2MKD-GDO 0004051 0619OOAD 6/19/00 23037.0 20019.2 1957@,FO,G5-3.2MKD-GDO 0004052 061900AD 6/19/00 36326.7 3 1567.9 19577F,FO,G5-3.2MKD-GDO 0004053 0619OOAD 6/19/00 19578F,FO,G5-3.2MKDGO 0004054 0619OOAD 6/19/0(3 34238.9 34808.0 29753.6 30248.2 19579F,FO,G5-3.2MHDGDO0004055 061900AD 6/19/00 25356.7 22035.0 1958OF,FO,G5-3.2MKpGDO 0004056 061900AD 6/19/00 24851.5 21596.0 AVERAGE 27524.1 239185 STANDARDDIEVIATION 4781 4155 Centre Analytical Laboratories,Inc. 3M Environmental Laboratory Page 26 of 103 Page 274 3NI 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 MI.Summary of PFOS residues in G1-Control on GD7 Sponsor CentreID Set Date Residue Found ID Number 1950lF.FO.G 1-Control-GD7 0004057 062000A 19502F,FO,G1control-GD7 0004058 062000A 19504F.FO.GlControl-GD7 0004059 062000A 1950.5F.FO.G 1-Control-GD7 0004060 062000A 19506F,FO,Gl-Co~0l-GD7 oO04061. 062000A 19507F,FO.Gl-Control-GD70004062 062000A 19508FP0,Gl-Control-GD7 0004063 062000A 195@F,FO,GlControl-GD7 o004064 062000A 195lOF,FO,Gl-ControI-GD7 0004065 062000A 19511F~O,G1Control-GD7 0004(166 062000AR 19512F,FO,G1Control-GD7 0004067 062000AR 19513F.FO,Gl-Control-GD7 0004068 062000A 19514F.FO,Gl-Control-GD7 0004069 062000A 19515F3O,G1-Control-GD7 0004070 062000AR Extracted .6/20/00 (ndg) ND NQ NQ hD 17.8 ND 0.3 MD ND ND ND ND ND ND 19516F3O,GlControl-GD7 0004071 062000AR 6/2OlOO NQ AVERAGE: 1.2 STANDARD DICVIATION 5 Corrected Residue Found (ndg) ND NQ NQ ND 15.5 ND 0.3 ND ND ND ND ND ND ND NQ 1.0 4 Centre Analytical Laboratories. Inc. 3NI Environmental Laboratory Page 27 of 103 Page 275 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor ProtocolNo:FAn-TOX-110 Table Vm. Summary of PFOS residues in G2-0.1MKD on GD7 Sponsor CentreID Set Corrected Date Residue Found Residue Found ID Number Extracted 19517F,FO.G2-0.1MKD-GD7 0004072 062100A 6/21/00 (ngg) 409.4 (n&) 355.8 19518F,FO,G2-0.1MKD-GD7 0004073 062100AD 6121/00 561.4 487.9 19519F,FO,G2-0.IMKD-GD7 0004074 062100A 6/21/00 344.6 299.5 19521F,FO,G2-0.IMKD-GD7 0004075 062100A 6/21/00 464.6 403.7 19522F,FO.G2-0.1MKD-GD7 0004076 0621QoAD 6/21/00 778.0 676.1 19523F,FO,G2-0.1MKD-GD7 0004077 062100AD 6/21/00 624.6 542.8 19524F,FO,G2-0.lMKD-GD7 0004078 062100AD 6/21/00 575.5 500.1 19525F,FO,G2-0.1MXD-GD7 0004079 062100AD 6/21/00 * 594.6 516.7 19526F,M,G2-0.1MKD-GD7 0004080 062100A 6/21/00 469.2 407.7 19527F,FO,G2-0.1MKD-GD7 0004081 062100AD 6/21/00 741.9 644.7 19528F,FO,G2-0.IMKD-GD7 0004082 062100AD 6/21/00 531.8 462.1 19529F,FO,G24.1MKD-GD7 0004083 062100AD 6/21/00 744.4 646.9 1953OF,FO,G2-0.1MKD-GD7 0004084 062100A 6/21/00 424.3 368.7 1953lF,FO,G2-0.1MKD-GD7 0004085 062100AD 6/21/00 623.9 542.2 19532F,FO,G2-0.1MKD-GD7 0004086 062100AD 6/21/00 573.9 498.7 AVERAGE: 564.1 490.2 STANDARDDEMATION 128 111 Centre Analytical Laboratories,Inc. 3NI EnvironmentalLaboratory Page 28 of 103 Page 276 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023416 Sponsor Protocol No: FACT-TOX-110 Table IX. Summary of PFOS residues in G3-0.4MKD on GD7 Sponsor CentreIO Set Corrected Date Residue Found Residue Found ID Number Extracted 19533F,FO,G3-0.4MKJI-GD70004087 062200m 6/22/00 (n,g) 2548.5 (ndd 2214.6 0004088 062200AD 6/22/00 2693.6 2340.7 0004089 062200AD 6/22/00 1824.8 1585.8 0004090 062200AD 6/22/00 3261.1 2833.9 0004091 062200AD 6/22/00 2611.1 2269.0 0004092 062200AD 6/22/00 2934.3 2549.9 0004093 062200AD 6/22/00 0004094 062200AD 6/22/00 2093.4 2267.8 . 1819.2 1970.7 0004095 062200A.D 6/22fOO 3003.0 2609.6 0004096. 062200AD 6/22/00 2797.7 2431.2 0004097 062200AD 6/22/00 2284.0 1984.8 ooo4098 062200AD 6/22/00 1523.1 1323.6 0004099 062200AD 6/22/00 2055.6 1786.3 0004100 062200AD 6t22f00 2847.6 2474.6 AVERAGE: STANDARD DEVIATION: 2481.8 492 2156.7 428 Centre Analytical Laboratories,Inc. 3NI Environmental Laboratory Page 29 of 103 Page 277 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 G4-1.6iMKD on GD7 Sponsor CentreID Set Corrected Date Residue Found Residue Found ID Number - Extracted 1955OF.FO.G4-1.6MKD-GD7 OOO4101 062200BD 6l2UOO (ndg) 14917.5 (ndg) 13015.4 19553F,FO,G4-1.6MKD-GD7 0004102 062200BD 6l2U00 12262.8 10656.4 19554F.FO,G4-1.6MKD-GD7 0004103 1955JF,FO,W1.6MKD-GD7 OOO4 104 19556F,FO,G4-1.6MKD-GD7 OOO4105 062200Bll 062200B21 062200BD 6/22/00 m2/w 6p22/00 11777.1 9607.3 10222.3 10234.3 8348.7 8883.2 19558F,FO,G4-1.6MKD-GD7 0004106 062200BD 6/22/00 5886.5 5115.4 19559F,FO,G4-1.6MKD-GD7 OOO4107 062200BD 6/22/00 6894.8 599 1.6 1956OF.FO,G4-1.6MKD-GD7 0004108 062200BD 6/22/00 7024.2 6104.0 . 19561F,FO,G4-1.6MKD-GD7 OOO4109 062200BD 6/22/00 8877.7 7714.7 19562F.FO,G4-1.6MKD-GD7 OOO4110 062200BD $/22/00 10845.1 9424.4 19563F,FO.G4-1.6MKD-GD7 OOO4111 062200BD 6/22/00 14910.3 12957.1 19564F,FO,G4-1.fjIvfKD-GDO7OO4112 062200BC 6/22/00 13656.0 11867.1 AVERAGE: 10578.5 9192.7 STANDARD DEVIATION: 3076 2673 Centre Analytical Laboratories,Inc. 3NI 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 SponsorProtocol No:FACT-TOX-110 Table XI. Summary of PFOS residues in G5-3.2- on GD7 Sponsor CentreID Set Corrected Date ResidueFound Residue Found ID Number Extracted l9565F,FO9G5-3.2MKD-GD7 0004113 062300AC' 6/23/00 (ng/g) 5 1657.5 (ndd 44890.4 1956@,FO,G5-3.2MKD-GD7 '0004114 062300AD 6/23/00 26453.8 22985.3 19567F,FO,G5-3.2MKD-GD7 0004115 062300AD 6/23/00 35913.2 3 1208.6 19568F,FO,G5-3.2MKD-GD7 0004116 062300AD 6/23/00 23007.4 19993.4 19569F,.FO,G5-3.2MKD-GD70004117 062300AD 6/23/00 42878.2 3726 1.2 1957OF,FO,G5-3.2MKD-GD7 0004118 062300AD 6/23/00 37368.9 32473.6 19571F,FO,G5-3.2MKD-GD7 0004119 062300AD 6/23/00 42995.9 37363.4 19572F,FO,G5-3.2MKD-GD7 0004120 062300AD 6/u/OO 38826.6 33740.3 19573F,FQ,G5-3.2MKD-GD7 0004121 062300AD 6/23/00 26685.9 23 190.0 19574F,FO,G5-3.2MKD-GD7 0004122 062300AD 6/23/00 36333.3 3 1573.6 19575F.FO,G5-3.2MKD-GD7 0004123 062300AD 6/23/00 35455.7 30811.0 19577F,FQ,G5-3.2MKD-GD7 0004124 062300AD 6/23/00 68339.2 59386.8 19579F,FO,G5-3.2MKD-GD7 0004125 062300AD 6/23/00 35856.5 31159.3 19580F.FO,G5-3.2MKD-GD7 ooo4126 062300AD 6/23/00 AVERAGE: STANDARD DEVIATION: 30065.5 37988.4 11516 26126.9 330113 10008 Centre Analytical Laboratories, Inc. 3ki EnvironmentalLaboratory Page 31 of 103 Page 279 3Nl 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 G1-Control on GD15 Sponsor CentreID Set Date Residue Found ID 19501F,FO,G1-Control-GDl5oOO4127 19502F,FO,Gl-ControI-GD150004128 19504F,FO,Gl-Control-GDl50004129 Number 062300B 062300B 062300B Extracted 6/23/00 6/23/00 6/23/00 (ne/g) hB hi IND 19505F~O,Gl-Control-GD150004130 062300B 6/23/00 IUD 19506F,FO,Gl-ControI-GD150004131 062300B 6/23/00 44.5 19507F,FO,G1-Control-GDl50004132 062300B 6/23/00 ND 19508F,FO,Gl-Control-GD15OOO4133 062300B 6/23/00 ND 19509F,F0,G1-Controlro1-GD150004134 062300B 6/23/00 24.2 195lOF,FO,Gl-C~trd-GD15 0004135 062300B 6/23/00 7.9 19511F,FO,G1-Control-GD150004136 062300B 6/23/00 ND 19512F,FO,G1-Control-GD15 0004137 062300B 6/23/00 57.9 19513F.FO,Gl-Control-GD15 0004138 062300B 6/23/00 ND 19514F.FO,Gl-Control-GD15 OOO4139 062300B 6/23/00 ND 19515F,FO,Gl-ControI-GD15 0004140 062300B 6/23/00 ND 19516F,FO,Gl-Cm@ol-GD15 0004141 062300B W23/00 21.6 AVERAGE: 10.4 STANDARD DEVIATION 19 Corrected Residue Found (ndg) 0.0 0.0 0.0 0.0 38.7 0.0 0.0 21.0 6.9 p.0 50.3 0.0 0.0 0.0 18.6 9.0 16 Centre Analytical Laboratories, Inc. 3NI Environ.mentalLaboratory Page 32 of 103 Page 280 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.:023-016 SponsorProtocolNo: FAn-TOX-110 Table XIII. Summary of PFOS residues in G2-O.1MKD on GD15 Sponsor CentreID Set Corrected Date Residue Found ResidueFound ID Number Exacted 19517F,FO,G2-0.1MKD-GD15 oooJ142 062600AI) 6/26/00 (ndg) 758.1 (np/g> 658.8 19518F,FO,G2-0.1MKD-GD15 0004143 062600AI) 6/26/00 19519F,FO,G2-0.1MKD-GD1O5OO4144 062600AI) 6/26/00 671.8 782.2 583.8 679.7 19521F,FO,G24.1MKD-GD15 0004145 062600ATI 6/26/00 19522FJ?O,G2-0.lMKD-GD105004146 062600Af) 6/26/00 19523F,FO,G2-0.1MKD-GDl50004147 062600Af) 6/26/00 632.3 817.4 641.7 549.5 710.3 557.6 19524F,FO,G2-0.1MKD-GD150004148 062600AD 6/26/00 . 19525F,FO,G2-0.1MKD-GDGD150004149 062600AD 6/26/00 834.2 1080.8 724.9 939.2 19526F,FO,G2-O.lMKD-GD105004150 062600AD 6/26/00 777.9 676.0 19527F,FO,G2-0.IMKD-GD150004151 062600AC1 6/26/00 798.5 693.9 19528F,FO,G2-0.1MKD-GD150004152 062600AD 6/26/00 844.8 734.1 19529F,FO,G2-0.1MKD-GDI05004153 062600AII 6/26/00 1953OF,FO,G2-0.IMKD-GD 15 0004154 062600AD1 6/26/00 723.6 724.4 628.8 629.5 1953lF,FO,G2-0.1MKD-GD15 0004155 062600AD1 6/26/00 777.3 675.5 19532F.FO,G2-0.1MXD-GD150004156 062600ADt 6/26/00 AVERAGE: STANDARD DEVWTION: 568.2 7623 119 493.8 662.4 104 Centre Analytical hbontories, Inc. 3kl Environmental Laboratory Page 33 of 103 Page 281 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.:023-016 Sponsor ProtocolNo:FACT-TOX-110 Table X V . Summary of PFOS residues in G3-0.4MKD on GD15 Sponsor CentreID Set Corrected Date Residue Found Residue Found ID Number Extracted 19533F,FO,G3-0.4MKD-GD105004157 062600BD 6/26/00 (n&) 2759.7 (ndd 2398.2 19534F,FO,G3-0.4MKD-GDl50004158 062600BD 6/26/00 4817.3 4186.2 19535F,FO,G3-0.4MKD-GD150004159 062600BD 6/26/00 3627.9 3152.6 19536F,FO,G3-0.4MJCD-GD150004160 062600BD' 6/26/00 19537F,FO,G3-0.4MKD-GD105004161 062600BD' 6/26/00 3186.8 3397.0 2769.3 2952.0 19538F,FO,G3-0.4MKD-GD150004162 062600BD 6/26/00 3143.5 273 1.7 19539F,FO,G3-0.4MKD-GDl50004163 062600BD 6/26/00 3094.1 2688.8 19540F,FO,G3-0.4MKD-GDl50004164 062600ED 6/26/00 4151.1 3607.3 19542F,FO,G3-0.4MKD-GDl50004165 062600BD 6/26/00 1881.6 1635.1 19533F,FO,G3-0.4MKD-GD150004166 062600BD 6/26/00 3420.1 2972.1 19544F,FO,G3-0.4MKD-GD150004167 062600BD 6/26/00 3372.6 2930.8 19546F,FO,G3-0.4MKD-GD15 0004168 062600BD 6/26/00 2625.1 228 1.2 19547F,FO,G3-0.4MKD-GD150004169 062600BD 6/26/00 3863.8 ' 3357.6 19548F,FO,G3-0.4hED-GD150004170 062600BD 6/26/00 3910.6 3398.3 -AVERAGE: 3375.1 , 2932.9 STANDARDDEVIATION 713 620 Centre Analytical Laboratories.Inc. 3fvl Environmental Laboratory Page 34 of 103 Page 282 3NI 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 CentreID Set Corrected Date Residue Found Residue Found ID Number Extracted 1955OF,FO,G4-1.6MKD-GDl50004171 062700AI) 6/27/00 19553F,FO,G4-1.6MKD-GD150004172 062700AI) 6/27/00 19554F,FO,G4-1.6MKD-GDl50004173 062700A.D 6/27/00 19555F,FO,G4-1.6MKD-GDl50004174 062700AI) 6/27/00 19556F,FO,G4-1.6MKD-GDl50004175 06270OAJl 6/27/00 19558F,FO,G4-1.6MKD-GD150004176 062700A.D 6/27/00 19559F,FO,G4-1.6MXD-GD105004177 062700All 6/27/00 1956OF,FO,G4-1.6MKD-GDl50004178 062700AJJ 6/27/00 1956lF,FO,G4-1.6MRD-GDl50004179 062700AC1 6/27/00 19562F,FO,G4-1.6MKD-GD150004180 0627WA.D 6/27/00 19563F.FO.W 1.6MKD-GDl5 0004181 062700Ac1 6/27/00 19564F,FO,G4-1.6MKD-G1D5 0004182 062700Ac1 6/27/00 AVERAGE: STANDARD D~EVIATION (ng/g) 13868.2 14838.7 14172.4 16258.1 9066.1 7174.9 7477.4 14880.9 17442.7 11417.2 8949.0 17342.4 12740.7 3769 (ne/& 1205 1.5 12894.8 12315.8 14128.3 7878.4 6235.0 6497.9 1293 1.5 15157.7 9921.5 7776.7 15070.5 11071.6 3275 Centre Analytical Laboratories, Inc. 3NI 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 GD1S Sponsor CentreID Set Corrected Date Residue Found Residue Found ID Number Extracted 19565F,FO,G5-3.2MKD-GDl50004183 062800AD 6/28/00 (ng/g) 35175.8 (n?k) 30567.8 19566F,FO,G5-3.2MKD-GD150004184 062800AD 6/28/00 19567F,FO,G5-3.2MKD-GDl50004185 062800AD 6/28/00 15192.6 20391.7 13202.4 17720.4 19568F,FO,G5-3.2MKD-GDlS0004186 062800AD 6/28/00 46897.0 40753.5 19569F,FO,G5-3.2MKD-GDlS0004187 062800A.D 6/28/00 36307.7 3 155 1.4 19570F,FO,G5-3.2MKD-GDl50004188 062800AD 6/28/00 31052.5 26984.6 19571F,FO,GS-3.2MKD-GDiS0004189 062800AD 6/28/00 28440.6 2471J.9 19572F,FO,GS-3.2MKD-GDl5OOO4190 062800AD 6/28/00 20549.6 17857.6 19573F,FO,G5-3.2MKD-GDl5OOO4191 062800AD 6/28/00 39763.7 34554.7 19574F,FO,G5-3.2MKD-GDl50004192 062800AD 6/28/00 47267.6 41075.5 19575F,FO,G5-3.2MKD-GD15 0004193 062800AD 6/28/00 40708.5 35375.7 19577F,FO,GS-3.2MKD-GDlS0004194 062800AD 6/28/00 47288.1 41093.4 19579F,FO,G5-3.2MKD-GDlS0004195 062800AD 6/28/00 33436.6 29056.4 19580F,FO,G5-3.2MKD-GDlS0004196 062800AD 6/28/00 32709.0 28424.1 AVERAGE: STANDARD DEVIATION 339413 10248 294953 8905 Centre Analytical Laboratories, Inc. 31vI Environmental Laboratory Page 36 of 103 Page 284 314 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 G1-Control on GD21 Sponsor CentreID Set Date Residue Found m l95O4F,FO,Glo~1trol-GD2 1 0004197 Number 062900A Extracted 6/29/00 (ndg) ND 19505F,FO.G1-Control-GD21 0004198 062900A 6/29/00 ND 19506F,FO,G1-Control-GD21 0004199 062900A 6/29/00 12.3 19507F,FO,GlControl-GD21 0004200 062900A 6/29/00 ND 19509F,FO,Gl-ControI-GD21Ooo4201 062900A 6/29/00 84.3 19512F,FO,Gl-Control-GD21 0004202 062900A 6/29/00 21.5 I9516F,FO,G1ControLGD21 0004203 062900A 6/29/00 2.0 AVERAGE: 17.2 STANDARD 1)EVLATION: 31 Corrected Residue Found (ndg) ND ND 10.7 ND 73.3 18.7 1.7 149 27 Table XVIII. Summary of PFOS residues in G2-O.lMKD on GD21 Sponsor Corrected Centre ID Set ' Date Residue Found Residue Found ID Number Extracted 19518F,FO,G2-0.1MKD-GD21 0004204 0629-3OOOA.D 6/29/00 19519F,FO,GZ-0.1MKD-GD21 0004205 062900A 6/29/00 1952lF,FO,G2-O.lMKD-GD210004206 062900A 6/29/00 19523F,FO,G2-0.lMKD-GD210004207 062900A 6/29/00 19524F,FO,G2-0.1MKD-GD210004208 062900A 6/29/00 (nglg) . 735.4 473.0 403.4 402.1 493.9 (n&> 639.1 411.0 350.6 349.4 429.2 19528F,FO,G2-0.1MKD-GD210004209 062900A 6/29/00 19529F,FO.G2-0.1MKD-GD;!l0004210 062900A 6/29/00 AVERAGE: STANDARD DEVIATION 456.1 - 391.4 4793 120 396.4 340.1 416.5 104 Centre Analytical Lnbontories, Inc. 3M EnvironmentalLaboratory Page 37 of 103 Page 285 3FA Medical Department Study: T-6295.12 I Analytical Report: FACT TOX-110 LRN42849 Centre Study No.: 023-016 SponsorProtocol No: FACT-TOX-1IO Table X N . Summary of PFOS residues in G3-0.4MKD on GD21 Sponsor m CentrelD Set Number Date Residue Found Extracted (ng/g) 19533F,FO.G3-0.4MKD-GD210004211 0629-3ooOAD 6/30/00 1684.4 19534F,FO,G3-0.4MKD-GD210004212 0629-3000AD 6/30/00 3102.9 19536F,FO,G3-0.4MKD-GD210004213 0629-3OOOAD 6/30/00 1848.1 19543F.FO,G3-0.4MKD-GD210004214 0629-3ooOAD 6/30/00 2233.1 19546F,FO,G3-0.4MKD-GD210004215 0629-30o0A.D 6/30/00 2229.2 19548F,FO,G3-0.4MKD-GD201004216 0629-3000AD 6/30/00 5417.3 AWRAGE: 27525 STANDARD DEVIATION 1395 Corrected Xejiiic: For?r.d (ndg) 1463.7 2696.4 1606.0 1940.6 1937.2 4707.6 23919 1212 Table XX. Summary of PFOS residues in G4-1.- on GD21 Sponsor CentreID Set Date Residue Found ID Number Extracted (nn/g) 19556F,FO,G4-1.6MKD-GD210004217 06293000AD 6/30/00 18272.7 . 19559F,FO,G4-1.6MIDGD21 0004218 0629-3000AD 6/30/00 10574.8 19560FJT),G4-1.6MKD-GD21 0004219 0629-3000AD 6/30/00 11190.1 19562F,FO,G4-1.6MKD-GD210004220 0629-3000AD 6MO/oo 5681.8 AVERAGE: 114299 STANDARD DEVIATION: 5185 Corrected Residue Found (ndg) 15879.0 9189.5 9724.2 4937.5 9932.5 . 4506 Table XXI. Summary of PFOS residues io G5-3.2MKD on GD21 Sponsor CentreID Set Date Residue Found ID Number Extracted 19568F,FO,G5-3.2MKD-GD210004221 0629-3000Al3 6/30/00 25084.0 19569F,FO.G5-3.2MKD-GD2 1 0004222 0629-3000AIl 6/30/00 21667.4 19571FJO,G5-3.2MKD-GD21 0004223 0629-3000A.I3 6/30/00 21465.5 19572F.FO,G5-3.2MKD-GD21 0004224 0629-3OOOA.I) 6/30/00 15006.4 19577F,FO,G5-3.2MKD-GD210004225 0629-3OOOAI) 6/36/00 27066.6 19579F,FO,G5-3.2h4KD-GD21 0004226 0629-3OOOAI) 6/30/00 28290.3 AVERAGE: 23096.7 STANDARD DE:VIATION: 4834 Corrected Residue Found Wd 21798.0 18829.0 18653.5 13040.6 23520.9 24584.3 20071.0 4201 Centre Analytical Laboratories, Inc. P3ge 38 of 103 3M Environmental Laboratory Page 286 3hA 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 Centre Set Date Amt. 56 1D ID Number Extracted Added (ng) Recovery 50lF,FO,G1-ControI-GDO 0003977 Spk A 061200AR 6112/00 50 126 WF,FO,G1-Control-GDO ooO3980 SpkB 061'200AR 6112/00 250 116 ~O5F,FO,Gl-Control-GDO 0003981Spk A 061400A 6/14/00 50 98 07F,FO,Gl-Control-GDO 0003983 Spk B 0614OOA 6/14/00 250 97 1lF,FO,Gl-ControlGDO 0003987SpkA 06lii0OAD 6/15/00 50 128 2F,FO,Gl-Control-GDO 0003988 Spk B 061fi00AD 6f 15/00 250 110 3F,FO,Gl-Control-GDO 0003989 Spk A 061600A 6/16/00 50 80 W,FO,Gl -Control-GDO 0003991SpkB 061600A 6/16/00 1000 89 .IFFO,G1-Control-GDO 0003986SpkA 061ZWAD 6/19/00 50 107 1 iF,FO,Gl-Control-GDO 0003987 SpkB 061SIooAD 6/19/00 5000 78 1 ~F,FO,Gl-Control-GDO 0003986Spk C 062000A 6/20/00 50 128 I! F,FO,Gl-Control-GDO 0003987 Spk D 062000A 6/20/00 2oooo 96 15 F,FO,Gl-Control-GDO 0003989Spk A 062100AD 6/21/00 1000 104 19 ?,FO,Gl-Control-GDO 0003991Spk B 062100A 6/21/00 2 m 80 19. 7,F0,G1-Control-GD7 0004057 Spk A 062200A 6/22/00 1000 97 19: '.FO,Gl-Control-GD7 0004060SpkB 062200A 6/22/00 2 m 79 195Cir. ],Gl-Control-GD7 0004062 SpkA 062200B 6/22/00 -1000 99 195C9F,FO,Gl-Control-GD'7 0004064SpkB 062200B 6/22/00 2 m 86 195lOF,FO,Gi-ControlGD7 0004065SpkA 062300A 6/23/00 1000 93 1951lF,FO,Gl-Control-GD7 oo04066 SpkB 0623CQA 6/23/00 2oooo 71 19512F,FO.Gl-Control-GD7 0004067SpkA %ZIOOB 6/23/00 loo0 85 19513F,FO,Gl-Control-GD7 0004068 SpkB 06ZJ00B 6/23/00 2oooo 81 19514F,FO,Gl-Control-GD7 0004069 SpkA 062ti00A m6/m 50 111 19516F,FO,Gl-Control-GD7 0004071Spk B 062ti00A a/?6/00 1000 94 l95OlF,FO,Gl-C~l-GD7 0004057 Spk C 06ZiOOB 6/26/00 50 82 19505F.FO,Gl-C0~trol-GD70004060 Spk D 062tDOB 6/26/00 2000 100 19501F,FO,Gl-Control-GD 15 0004127 Spk A 062i'OOA 6/27/00 100 100 19502F,FO,G1-Control-GDl50004128SpkB 062700A 6/27/00 zoo00 100 19504F.FO,G1-ControlGD15 0004129Spk A 0628OOA 6/28/00 500 99 19505F,FO,G1-Conml-GDl5 0004130Spk B 062800A 6/28/00 1OOOOO 89 19507F,FO,G1-Control-GD15 0004132SpkA 0629.00A 6/29/00 50 119 19508F,FO,Gl-Control-GD15 0004133 Spk B 062900A 6/29/00 ZOO0 95 19513F,FO.Gl-Control-GD15 0004138 SpkA ' 063000A 6/30/00 500 113 19514F~O,G1-Control-GD15 0004139 Spk B 063000A 6/30/00 2oooO 85 AVERAGE: 98 STANDARD DEVIATION 15 RELATIVESTANDARDDEVIATION 15 Centre Analytical Laboratories. Inc. Page 39 of 103 3M EnvironmentalLaboratory Page 287 3PA Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 . Centre Study No.:023-016 Sponsor Protocol No: FACT-TOX-I 10 12.0 - FIGURES Centre Analytical Laboratories, Inc. 3M EnvironmentalLaboratory Page 40 of 103 Page 288 3M Medical DepartmentStudy: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Figure 1. Cenrre Study No.:023-016 Sponsor Protwo1 No:FACT-TOX- 110 Typical Calibration Curve for PFos , 50.0 ' ioo.0 I ' ' 150I .0 ' 20b.0 ' ' I nglmL 250.0 CentreAnalytical Labontories.Inc. 3M EnvironmentalLaboratory Page 41 of 103 Page 289 ~~ 3FA Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Figure 2. Centre Study No.: 023-016 Sponsor PrOtccOl No: FAm-TOX-110 Typical Mean Response Factor for THPFOS CO~pound2 name: THPFOS Response Factor: 407.31 3 Response type: External Std, i r e a Surve type: RF ' 1.09e x X . X . .. .. Responsc -c ' ' 4 . 50.0 i5b.b I ' ' 1ob.o ' ' ' ' ' I" ' ' ' .. 200.0 . . ing/mL 250.0 Centre Analytical Laboratories, Inc. 3M EnvironmentalLaboratory Paze 42 of 103 Page 290 3hl Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN42849 Figure 3. Centre Study No.:023-016 Sponsor ProtocolNo: FACT-TOX-I10 ChromatogramRepresenting a'5 n g / a extracted standard for PFOS and 250 n g h L fortification of THPFOS 0612OoAR-2002 Sm (SG, 2x2) I 2: THBFOs ~CO61200-1,5ng/mL Standard j iI 1 I "i I 0 .(, 1.00 5.11 923071 I. . ,.b'. '2.00' " 3.00 ' 4.00 ~._ %/ 1 3 4 ~ ~ - 2 0 0090:oo: LCIMSlMS MRM of2 Channels ES- . 499 7 gd 6.24e41 Areai . ' 7.00 ' 8.06 ' Time 13-Jun-2L0C0I0M0S9l:M00S:2#< MRM of 2 Channels ES 427 > a1 8.67e: Are; Centre Analyticalhlbontories, Inc.. 3M EnvironmentalLaboratory Page 43 of 103 Page 291 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 ' LRN-U2849 :- ;s C 1200-5,125 ng/mL Standard '1 'AR-2006 Sm (SG, 2x2) % 5.26 ~ I101957 1 3.00 ' 4.b0 ' 5.00 -r 13Jun-2000 10:07:191 I LClMSlMS $6 MRM of 2 Channels ES- 4*-Y-Y e 4 8.76e5 Ared Centre Analytical Labontories. Inc. 3M EnvironmentalLaboratory Pqge 44 of 103 Page 292 3hl Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Figure 5. Centre Study No.: 023-016 SponsorProtocoI No:FACT-TOX-1IO Chromatogram Representing Control Rat Feces for PFOS m dTHPFOS (Centre m:0004277 Blank B, Set: 062300A) 5.61 499 > 09 3.11e4 , . a , . Time LCIMSIMS $61 Centre Analytical Laboratories,Inc. 3M Environmental Laboratory Page 45 of 103 Page 293 3NI Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Figure 6. Centre Study No.:023-016 Sponsor Prctccol No:FAfl-TOX-I10 ChromatogramRepresentjing Control Rat Feces Fortified with 50 ng of PFOS and 500 ng of THPFOS (Centre ID:0003951 Spk A., Set: 061400.4) :61400A-1013Sm (SG. 2x2) -- 6.00 ' MRM of 2 Channels 427 ESa-o 7.84e5 Area ...,. . I . . . . Time 7.00 . 8.b; ' Centre Analyticd hboratories, Inc. 3NI EnvironmentalLaboratory Page 46 of 10; Page 294 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Figire 7, Centre Study No.:023-016 Sponsorh t o c o l No:FACT-TOX-110 Chromatogram of Rat Feces Sample from G2 on GDO (Centre ID: 0003994, Set.: 061400A) .I 1: PCOS 0003994 . 061400A-1018 Sm (SG. 2x2) . . -. , , 5.24 138530 . 14-JUn-2000 23:32:3$ LCNS/MS*6i : MRM'of2 Channels ES- 061400A-1018Sm (SG, W ) LC/MS/MS $6 MRM of 2 Channels ES427 > e0 Centre AnalyticalLabontories,Inc. , 3M EnvironmentalLaboratory Page 47 of 103 Page 295 3NI Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No:FACT-TOX-I 10 13.0 APPEINDICES Centre Analytical Laboratories, Inc. 3NI EnvironmentalLaboratory Page 48 of 103 . Page 296 3h4 Medical DepartmentStudy: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.:023-016 SponsorProtocol No:FACT-TOX-110 APPENDIX A Study Protocol FACT-TOX=llO (Centre Study No. 023-el6) .- and Amendments and1 Deviations Centre Analytical Labontories. Inc. 3M EnvironmentalLaboratory Page 49 of 103 Page 297 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U. 28.4..9-.-... I Study Title , Oral (Gavage) PhmmAchetiC Study of PFOS in .. PROTOCOL . Author Lisa CIcmu - Date: June 8,1999 -. Perfoming Lahorafoty 3ME ~ ~ I O I U I ITUe=I h~n~o~Iog~8r Safety Services .3M EnVtpIlmcptal Laboratory 9 3 Bush Avmue , StP3uZMN 55106 Laboratory Projecfj d e n t i f i t w h ' FACT-TOX-I 10 u2949 Centre Analytical. Laboratories, Inc. 3M Environmental Laboratory . .. . . i' Page 50 of 103 Page 298 3111 Medical Department Study: T-6295.12 .-Test Material -- Analytical Report: FACT TOX-110 I .LRN-U2849 . . . . .. Centre Study No.: 023-016 --___ SponsorProtocol No:FACT-TOX-110 . ...- . ._ . ... . .. -._ . . .. ..-..-._.- Sponsor Representalive .. Study Director - -- Study Location(s) in vivo TesfingFauW MarvinT.Case,D.VX, Pm. 3M Toxiwlo!~Sewicx Telephone: 651-735-5180 F a d = 63 1-733-1773 . Centre Analytical Labntories.Inc. 3M Environmental Laboratory Page51 of 103 Page 299 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Ceatre StudyNo.:023-016 SponsorProtocol No:FACT-TOX-110 . Proposed Study Emetable Study Inifiatian Dafe Study CampleiionDate .. June 8.1999 ' August 8,2.(.xHl . . . 3M E M m e n i a l - .......- ". . . . , . Cen.tre!AnalyticalLa.b.ora.t.o.ries,Inc. . 3NI EnvironmentalLaboratory pa& 3of ju Page 300 -- 3bl Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 . . ... . . , ._. .... ., . . _-" _. . .._..-. ---.-.-.-..- . Centre Study No.:023-016 Sponsor Protocol NO: FACT-TOX-LIO ... . . . .. . . --.-. --- .-..-.__.-... . CentreAnalytical Laboratories,Inc. .... . . .... . . 3M EnvironmentalLaboratory Page 53 of 103 Page 301 3M Medical Department Study: T-6295.12 .. 7.8 Analytical Report: FACT TOX-110 LRN-U2849 . . .. - Centre Study NO.:023-016 . SponsorProtocolNo:FACT-TOX-110 ' -.. . ...I... ._......._-I-.-- PIG&I %ACT-T~--0:, . I .Table 2 Specimen Information 3rd EnvironmentalLaboratory Page 54 of 103 Page 302 3h4 Medical Department Study: T-6295.12 . .. , . _.._. .. Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Spo-nsor Protocol No: FACT-TOX-110 . ......e ...__-.-.-.-..-.- - --. _.. Centre.Aqdyt..id_L-a..b.or.atories, Inc. 3M Environmental Laboratory .. Page 55 of 103 Page 303 3NI Medical Department Study: T-6295.12 12.7 Linearity 2 2 0.98 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 . . ...-...-. -I-. . ._.... ..._. Centre Analytjcal hbontories,,Inc. 3M Environmental Laboratory Page 56 of 103 Page 304 3bl Medical Department Study: T-6295.12 I Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 __ -__- .. ,, , ... . .. SponsorProtocol No:FACT-TOX- 110 . ....... - - , . .... .. . - .-...._ ..-.--*-."_--.--..I. Centre Analytical Laboratories, Inc. 3M EnvironmentalLaboratory Page57 of 103 Page 305 3kl Medical Department Study: T-6295.12 i Analytical Report: FACT TOX-110 LRN-U2849 ...... 3M bvircnmentd Laberafy, .Centre AnaiylicalLaboratories, Inc. 3M Environmental Laboratow .. Pagd 9 cf IO .j Page 58 of 103 ' . Page306 -- 31\41Medical Department Study: T-6295.12 -- Analytical Report: FACT TOX-110 LRN-U284.9.. Centre Study No.:023-016 Sponsor Protccol No:FACI'-TOX-I 10 . . .-. 3 4 Environmental . - . . . . Centre Andytical Laboratorie,s.,..h.f. 3M Environmental Laboratory Page IO c? :a Page 59 of 103 .. .. , .. ._ . Page 307 3Fd Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.:023-016 Sponsor Protccol No: FACT-TOX-110 .. ..-- -.. .. Study 77Ue (~avrge)P ~ , ~ u x & u ~Sctudy of&OS iaRats ' PROTOCOL AMENDMENTNO. 1 Amendment Date: Aqust 12,1999 -. Laboratory ProjectIdenUficat:on ET&SS FACT-TOXI IO URN u2s49 . , ... Centre Analytical Laboratories.Inc. . .._. 3M EnvironmentalLaboratory .. Page 60 of 103 Page 308 3hA Medical DepartmentStudy: T-6295.12 CF-- I . Analytical Report: FACT TOX-110 LRN-U2849 __..--. . ,.. .. .. Czntre Study NO.:023-016 Sponsor Protocol No:FACT-TOX-110 .-..- .--.-.,..-.. .. Centre Analyticai Laboratories,hc. 3M EnvironmentalLaboratory . .. .. Page 61 of 103 Page 309 _- 3hA Medical Department Study: T-6295.22 . .,. . . . __.&. Analytical Report: FACT TOX-110 LRN-U2849' ...... " ...-..... . . ... . . . . . .A" . . . . . . . . . ...:..i . . . . . . . . . . . . . . . . - Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Centre Analy$cal Laboratories, hc. 3M Environmental Laboratory Page'62of 103 ' Page 310 3hl Medical Department Study: T-6295.12 .. .. .- .. . . . .. .. ._.. .. . - Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protccol No: FACT-TOX-I 10 . .. .. ...._ Centre Analytical Laboratories. Inc. . 3M EnvironmentalLaboratory Page 63 of 103 . Page 311 3hl Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 .. . . Centre Study No.: 023-016 Spo-nsor Protocol No: FACT-TOX-110 .. . Study Title Oral (Gavage) PharmaurkincticSt?dy ofPFOSin Rats PROTOCOLAMENDMENT No.2 -Amendsent Date: Septcmbe:Z8,1999 .. . .- Laboratory Pmject ldentiiication E T B S FMT-TOXI 10 LIRN ms49 . .. ,: . . Centre AnalyticalLab.ora..tories,Inc. 3M EnvironmentalLabdratory PGge'64 of 103 . Page 312 3h4 Medical Department Study: T-6295.12 .. . .. .. . Analytical Report: FACT TOX-110 LRN-U2849 .... I , Centre Study No.: 023-016 Sp-o'n.sor Protocol No:FACT-TOX-110 Centre ,&alytical Laboratories, hc. 3M Environmental Laboratory .. Page-65 of 103 Page 313 3M Medical Department Study: T-6295.12 ... Analytical Report: FACT TOX-110 LRN-U2849 cent^ Study NO.:023-016 __ Sponior Protocol No: FACT-TOX-110 ... .. I. ' ." .. --._-... ...- ._-- .. Centre Andytical Laboratories. Inc. 3M Environmental Laboratory .. Page 66 of 103 .. .. .. Page 314 3hl Medical DepartmentStudy: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 ...... Centre Study No.:023-016 _-- ... .....S-...p..o-n--.s_or..Pr.o.to. .c.o.l.N. o. :..F-A--m-.--T.OX-1 IO .. . Amendment AppravaI . . . . Pakw MCT-TOXI o, . heccrnent No. 2 . .. .. .. .- ... ...-.-- --.-.. ..Centre Analytical L.ab..o.-r_a.t-o..ries. Inc, ... 3M Env/ronmentalLaboratory .- .. , . .. Page 67 of 103 , .. Page 315 3h9 Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 I . .. Centre Study No.:023-016 Sp. on-sorProtccol No:FACT-TOX-110 .. .. i PROTOCOL AMENDMENT NO. 3 ' Amendmant Date: . 20 January 2000 . . e . Laboratory Project Identif7catio.n ET&ssLRN-m49 . FACTTOX-I 10 A r g ~ ~ S t u d y41: s-013 j&[>[&A D m t SUdy: T-6295.12 .._. .. ..._..Centre AnalyticalLaboratories,he.. 3M Environmental Laboratory Page 68 of 103 : Page 316 3M Medical Department Study: T-6295.12 --- Analytical Report: FACT TOX-110 LRN-U2849 .. . . Centre Study No.:023-016 ' Spon-sor Protccol No:FACT-TOX-110 .. . ..,- Pnt- w - u z a 4 9 Amendment Nmber 3, Centre AnalyticalLaboratories,Inc. , ' - 3M Environmental Laboratory .. Page'69 of 103 : Page 317 3111 Medical Department Study: T-6295.12 . . . .. .. . Analytical Report: FACT TOX-110 ... LRN-U2849 Centre Study No.: 023-016 Spwsor Protwo1 No: FACT-TOX-110 . ...-.---.'- . Amendment Approval i . .. T D.V%PhD, IncomingStudy Dkctor Centre h a l y t i c dhbontories,hc.. . 3M EnvironmentalLaboratory Page'70 of 103, Page 318 3lbl Medical Department Study: T-6295.12 .. i Analytical Report: FACT TOX-110 I LRN-U2849 Centre Study No.:023-016 - Sponsor Protocol No: FACT-TOX-I 10 .. . . ...... --.__.. PROTOCOL AMENDMENT NO,4 .. .. . Amendment Date: . 20 Apd 2000 Perfomring Laboratories and Satcty S m i c t r LaboratoryProject Idenlifeation E T B S LRN-uzs49 FACT TOX-I IO ' ArpStudy: 418013 2,Cf bfedid Ilcptmat S e u k T-6295.12 . . . -...... .. Centre AnaIytical h.boratories.Iac. 3M Environmental Laboratory Page 71 of 103. Page 319 3M Medical Department Study: T-6295.12 I . I Analytical Report: FACT TOX-110 LRN-U2849 . ... . ~ Centre Study No.: 023-016 - Sponsor Protocol No: FACT-TOX-1 IO .. . ..._. Centre Analytical hbontories, Inc. . 3M Environmental Laboratory . Page72of 103 Page 320 3rd Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.:023-016 .. Sponsor Protccol No: FACT-TOX-I 10 .' Amendment Approvcl 1 .. .. Centre Analytical Laboratories, Inc. . . 3M Environmental Laboratory Page 73 of 103: Page 321 3h4 Medical Oepartment Study: T-6295.12 Analytical Report: FACT TOX-110 i LRN-U2849"- Cenwe Study No.:023-026 Sponsor Protocol No: FACT-TOX-110 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 74 of 103 Page 322 3PA Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849' . Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-I 10 ..-..-. .--". . Centre Analytical Labovtories,Inc. 3M EnvironmentalLaboratory Page 75 of 103 Page 323 3hA Medical Department Study: T-6295.12 I . .. Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor ProtocoI No: FACT-TOX-110 .APPENDIX B Determination of Fluorochemical Residues in Monkeymat Feces by Lc/Ms/Ms (Revision 2) Method #00M-023-003, revision 2'and Deviations .and Modifications Centre Analytical Laboratories, Inc. 3M EnvironmentalLaboratory Page,76of 103 Page 324 3Ftl Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 . . ..._ -.. Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-I10 .. AUTHORS T0T.U NGMEER OF PAGES 20 Centre Analytical Laboratories,hc. ' 3M Environmental Laboratory ' .. . Page 77 of 103 Page 325 3hA Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 Sponsor Protocol No: FACT-TO,X-lIO . Centre Analytical LabontoriM, I n c a . .... . ... .. _.. 3M EnvironmentalLaboratory Page 78 of 103 Page 326 9- .. Centre Study NO.:023-016 SponsorProtocol No:F.4CT-TOX-110 . ..._...-. 4. .. Page 79 of 103 ' page327 3pn Medical Department Study: T-6295.12 .. Analytical Report: FACT TOX-110 LrRN-UZ844- ..... ....---._ . I Centre Study No.:023-016 Sponsor ProtocoI No:FACT-TOX- 110 . .- - ,_. ,i ,.. .. Centre Analytical Laboratories,hc. .. .. . . .... .., . .. . 3M EnvironmentalLaboratory Page 80 of 103 Page 328 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 -. PFOSD Molrmlar weight 527 Centre Analytical Laboratories, Inc. , 3M Environmentaliaboratory Page 5 Page41 of 103 Page 329 3CA Medical Department Study: T-6295.12 I Analytical Report: FACT TOX-110 LRN-U2849 . . ... Centre Study No.:023-016 Sponsor Protocd No: FACT-TOX-1 LO . . . .... .-.- Centre h d y t i e d Laboratories, Xnc. .< 3M EnvironmentalLaboratory Page 6 Page 82 of 103 Page 330 3h4 Medical Department Study: T-6295.12 3. ' Analytical Report: FACT TOX-110 . LRN-U2849 .. . ... ... -.-.-- . .... Centre Study No.:023-016 . Sponsor ProtocolNo:FACT-TOX-1IO 3.2. FLUOROCHEMSITCAANLDAW$ Standard PFOS PFOSA PFOSAA PFOSEA Et-FOSE-OH Mj56 M570 POAA ' 'THPFOS SOW7 ;JAY 3M' 34 . 3M 3M 3M - 3 4 3M 3rM 3.3. EQUIPMENTAM)SUP~ Centre Analytical Laboratories,hc. 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 yo: FACT-TOX-110 .. ........ Centre AnalyticalLaboratones,Inc. 3M Environmental Laboratory Pigc 3 Page 84 of 103 .. Page 332 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study Na.:.023-016 Sponsor Protocol yo: FACT-TOX-110 Centre hdyticd Laboratories, Inc. 3M EnvironmentalLaboratory Page 85 a 103 ' Page 333 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.:023-016 SponsorProtocol No: FAn-TOX-1 IO . . . . . . . . . .-.. Centre Analytical Laboratories,Inc. ...- . . . . . . . . . 3M Environmental Laboratory . Page 86 of 103 Page 334 3M Medical Department Study: T-6295.12 .. -. 4. METEOD Analytical Report: FACT TOX-110 LRN-U2849 --. . .. .. Czntre Study No.:023-016 Sponsor Protocol NO: FACT-TOX-110 ...._..._..___. 4.3. SAMPLPEm.m'TION - . _.. . Centre Analytical Laboratories.Inc. . . . . .. 3M Environmental Laboratory Page 87 of 103 Page 335 3M Medical Department Study: T-6295.12 , AnalyticalReport: FACT TOX-110 LRN-U2849. ~ Centre Study No.: 023-016 Sponsor Protocol No: FACT-TOX-110 Centrehalytical Laboratories, Inc. .. 3M EnvironmentalLaboratory Page12 ' Page 88 of 103 Page 336 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 . .Centre'StudyNo.: 023-016 Sponsor Protocol NO:FACT-TOX-I10 . ._ . . ._.-. 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 I LRN-U2849 ........ - .. , _. . ' Centre Study No.: 023-016 SponsorProtocol No:FACT-TOX-110 ............ ..... -_-.-.-. & #A 0 60 0.4 60 1.0 7.0 75 9.0 93 123 14.0 ..-... . . . . . . . . . . Centre halytical Laboratories.hc. 3M EnvironmentalLaboratory Flow Rate C-m) O2W ' 0200 , os00 0200 0200 0.400 0.400 o.Jao 0200 Page.90of 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-110 , . -- .. .. ..._... . i F 0 S PFOSA PFOSAA PFE t:FOSE-OB POAA a M556 M570 THPFOS - Mode - Pmntloa 499 498 581 526 650 413 556 570' 427 Product Approhate rOn ' Retention ~ j m (emin) 99 53 78 6.0 526 5.7 I69 6.7 59 * 6.7 369 5.1 498 5.4 419 5.6 80 5. I && PFOS - D k l I fd . 0.2 0.2 28 0.2 20 02 25 0.2 51 0.2 11 02 28 0.2 20 0.2 35 Centre haIytical Laboratories,Inc. , . - .. 3M Environmental Laboratory Cone 76 34 57 49 30 25 50 60 34 Page IS Page 91 of 103 Page 339 3M Medical Department Study: T-6295.12 I .. Analytical Report: FACT TOX-110 LRN-U2849 C~ntreStudy NO.0:23-016 SponsorProtccol No:FACT-TOX-110 WW Hc~poPoleI Apeime I Hl?.qmle 2 Source Block Tmp. DaolradonT a p . Prmma rJa5Jcdl -Sd 3-56 w 0.5 v 02 v 0.8 v 1000c 'W C 13.0V a7 v -2 Y 1v ll.0V 1I.OV 1.0 v 650 Y Centre AnalyticalLaboratories,Inc. 3M Environmental Laboratory h p c 16 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 SponsorProtocol No: FACT-TOX-110 .- .. .. ... .. . .. . Centre Analyticalhbontories, hc. .. . 3M EnvironmentalLaboratory Page 17 Page 93 of 103 Page 341 3M Medical Department Study: T-6295.12 .. . . .. Analytical Report: FACT TOX-I 10 LRN-U2849 ' I Centre Study No.: 023-016 Sponsor Protccol No:FACTrTOX-I10 5. CALCULATIONS 5.1. AVALYIZFOUND Equation 1: .. . -.. ... .. Centre Analytical Laboratories. Inc. . .. 3M Environmental Laboratory Pqc IS Page 94 of 103 Page 342 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN.-U..2-8-.4...9.. Ccme Study No.:Ok-016 Sponsor P:otccol No:FACT-TOX-110 5.3. MEANREVONSE~A~~OR . 6. SAFETY . . . . . . . . . . . . . Centre Analytical hbomto' ti.es, h.c. 3M Environmental Laboratory Page.95 of 103 Page 343 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 Centre Analytical Laboratories, Inc. 3M Environmental Laboratory I I 1 Pqe 20 Page 96 of 103 Page 344 3M Medical Department Study: T-6295.12 I .. Analytical Report: FACT TOX-110 L. .R..N..-.U..2. .8.4,9. Centre Study No.:023-016 Sponsor ProtocolNo:FAa-TOX-110 .. DESCR~P~OONF DEVIATION Centre Analytical Laboratories,hc. 3M EnvironmentalLaboratory Page 97 of 103 Page 345 3M Medical DepartmentStudy: T-6295.12 - .... . . Analytical Report: FACT TOX-110 ... LRN. -.U...284._9._. Centre Study No.:023-016 -Sponsor Protocol No: FACT-TOX-I10 , . .. METHOD DEVIATION Devi.uionNu~r~k1 Ps5c 2 or 2 .. Centre AnalyticalLaboratories. Inc. . . - ... 3M EnvironmentalLaboratory Page 98 of 103 ' Page 346 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 . ,. . ...... .. Centre Study No.:023416 Sponsor Protocol No: FACT-TOX-110 -. . CentreAnalytical 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 SponsorProtocol No:FACT-TOX-I 10 Centre Analytical Laboratories, Jnc. 3M EnvironmentalLaboratory - Page 100 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-I 10 Method .D(Reetevrimsioinna2ti)onCofcFmluMorectchhoedmNicudpRbeesri:d0u0eMsh-oMudOo3n.krceyvmisikonc2s by L.CIMSMS UODFICATIOW, Section 3.4 Solutions 2. The 100&I a~emtesolution should be 50 mMYnmouniumaccctate. flSTIFIC4TION: To correct foea typognphica~error. No negative -impax. Centre Analyticai Laboratories,Inc. 3M EnvironmentalLaboratory 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 Laboratori.e.s.,.I.nc. . 3M Environmental Laboratory Page 102 of 103 Page 350 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 Centre Study No.: 023-016 SponsorProtocol No: FACT-TOX-110 . ... METHOD MODIFICATIONFORM Pap 2 of 2 Centre Analytical Laboratories,Jnc. 3M Environmental Laboratory Page 103 of 103 Page 351 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 - \ inc, CEntrE Analgtical Laboratories, 3048ResearchDdve, State College, PA 16801. Phone: (814) 231-8032, Facsimile: (814)231-1253 21 Amended pages + amendmentform ANALYTICAL PHASE REPORT AMENDMENT Amendment Number: 1. Effective Date: 9/20/00 Sentre 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: 59386.8ppb was changed to 59.4 pglg in the third paragraph (Seeattached Pagel 1). 3ection 7.6 Quantitation and Example Calculation: Equation 5: Residue Found (pglg) =ResidueFound (nglg) * (1pg / lo00ng) Equation 6 Corrected Residue Found (pg/g) = Residue Found (pg/g) * 0.869 rhese equations were only applied to the tables in the report. Section 8.0Results: 59386.8was changed to 59.4 pg/g in the second paragraph (See attached page 18). . Section 11.0: Tables I- XXI (See attachedpages 20- 38) rhe 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 11and.Xrr were changed to ND's, and any value less than 0.0100 pglg (LOQ) was reported as < 0.0100. No negative impact /,I\ h p&fb/--.J!L\*, ; Impact on the Study Sddy Director Signature CAL QAU Review E/cc- Date qoLf Date 1IzT l p y 9H-Q November 199510 3M Environmental Laboratory Page 352 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 . . LR. _N-_ U2-_8_4_ .9._..-._.. Centre Study No.: 023-016 SponsorProtoFol No.:FACT-TOX-110 1.0 SUMMARY The purpose of this study was to anaIyze residues of perfluorooctanesulfonate(PFOS) in rat feces as specified in 3M Protocol FACT-TOX-110. The adytical method used for this study was entitled, "Determination of Fluorochemicd Residues in Monkeymat Feces by LCIMSMS, revision 2" (Centre method number: OOM-023-003, revision 2). The limit of quantification for PFOS in rat feces was 10 ppb (0.0100 pg/g). Residues (corrected for purity) ranging from nondetected levels to 59.4 pg/g were found in the rat feces samples. Fofiification recoveries ranged fro? 71-12896 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 Monkeymat Feces by LUMSMS, revision 2." .- 3.0 INTRODUCTION This report &tails 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 LWs/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-110Amendment #4. The complete prOtOcO1 and amendments can be found in Appendix A. The analyticalstart date was June 9,2000,and the analyticaltermination date was July 1,2000. 4.0 TESTSYSTEM The 250 rat feces sariples analyzedin 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 forstandards and blanks W;IS p&hased from Lampire ' Biological L&ontories, Inc., Pipersville, PA and received at Centre frozen on dry ice on 3M Environmental Laboratory Amended Page 11 of 103 Page 353 3 M Medical Department Study: T-6295.12 I Analytical Report: FACT TOX-110 LRN42849 - Centre Study No.: 023-016 SponsorProtocol No.:FACT-TOXh 10 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 mahix 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 fie PFOS standard lot 217 based on the certificate of q'ialysis findized on September 7, 2000. Fortification recoveries ranged from 71-12896 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 THATMAY HAVEAl?F'ECTED 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 instrumentiltion 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 assufilIIce. Verified exact copies and the eleCtrOnic data have been stored.in the archives at Centre Analytical Laboratories, %IC. 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; h c . 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 dl original facility-specific raw data,will be retained in the Centre Analfical Laboratories, Inc. archives for the period of time specified in 21 CFR Part 58. Retained samples of reference substances are archived by the sporisor. 3 M Environmental Laboratory AmendedPage 18of 103 Page 354 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 _- L.R.N- -lJ2849--- Centre StudyNo.:023-016 I Sponsor Protocol No.: FACT-TOX-110 I Table I. Summary of PFOS residues in Matrix Blanks and Matrix Zero Blanks ND =Not Detected and NQ =Not Quantifiable i Sponsor Centre Residue Corrected Set Date Found Residue Found ID ID Number Extracted (pglg) WE) na 0004277Blank A 061200AR 6/12/00 ND ND na 0004277BlankB 061200AR 6/12/00 ND na 0004277Zen,Blank C 061200AR 6/12/00 ND na 0004277Zero Blank D 061200A.R 6/12/00 ND na 0004277Blank A 061400A 6/14/00 NQ na 0004277Blank B 061400A w1400, NQ na 0004277Zero B l q k C 0614OOA ' 6/14/00 NQ na 0004277ZeroBlank D 061400A 6/14/00 ND na 0004277 Blank A 0615OOAD 6/15/00 NQ na 0004277 Blanlr B 061500AD 6/15/00 NQ na 0004277Zero Blank C 061500AD 6/15/00 < 0.00869 na 0004277Zero Blank D 061500AD 6/15/00 < 0.00869 na 0004277Blank E 061600A 6/16/00 < 0.00869 na 0004277Blank F 061600A W16l00 NQ na 0004277 Zero Blank G 06LMXIA 6/16/00 NQ na . 0004277ZemBlankH 061600A 6/161M) NQ na 0004277BlankA 061900AD 6/19/00 NQ nil 0004277BlankB 0619OOAD 6/19/00 NQ na 0004277Zero Blank C 0619OOAD 6/19/00 NQ na 0004277Zero Blank D 061900AD 6/19/00 NQ na 0003466 Blank A 062000A 6/2WOO na 0003466Blank B 062000A 6/2WOO NQ na 0005466Zero Blilak C 062000A 6/2o/oo na 0003466 Zero Blank D 06U)oA 6/2W# na 0004277 Blanlt A 062100A 6/21/00 na 0004277 Blank B 062100A 6/21/00 na 0004277ZeroBlank C 062100A mlm na 0004277Zero Blank D 0621OOA 6/21/00 na 0004277 Blank A 062200A 6/22/00 na 0004277BIank B 062200A 6/22/00 oa 0004277Zero Blilnk C 062200A 6/22/00 NQ NQ NQ NQ NQ NQ < 0.00869 ND ND na 009277Zero BlankD . 062200A 6/22/00 ND na 0004277 Blank E 062200B 6/22/00 m na 0004277 Blank F 06wMB 6/22/00 ND na 0004277 Zero Blank G 0622WE 6l22lciQ ND 3M EnvironmentalLaboratory Amended Page 20 of 103 Page 355 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U.2.84_8-.--_. . -ental ! 1 ! Centre Study No.: 023-016 , Sponsor Protocol No.: FACT-TOX-110 Table I (cont.) Summary of PFOS r&idues in Matrix Blanks and Matrix Zero Blanks ND = Not Detected and NQ = Not Quantifiable Residue Corrected Residue Sponsor ID na na na na Centre ID m277ZeroBlankH 0004277Blank A 0004277 Blank B 0004277 Zero Blank C Set Number 062200B 062300A 0623OOA 062300A Date Extracted 6/22/00 6/23/00 6/23/00 6/23/00 Found (pe/g) ND MI ND ND Found (l.ldg) ND ND ND ND na 0004277 Zero Blank D 062300A 6/23/00 ND ND na 0004277 Blank A 06U00B 6/23/00 NQ NQ na 0004277Blank B 062300B 6/23/00 NQ NQ na 0004277ZeroBlank C 062300B 6/23/00 NQ NQ na 0404277 ZeroBlank I) 062300B 6/23/00 NQ NQ na 0005738 Blank A 062600A 6/26/00 < 0.0100 < 0.00869 na 0005738BlankB 062600A 6/26/00 0:0126 0.0109 na 0005738 Zero Blank C 062600A 6/26/00 ND ND na 0005738ZeroBlank D 062600A 6/26/00 N D ND na 0005738Blank E 062600B 6/26/00 ND ND na 0005738 Blank F 062600B 6/26/00 N D - ND na 0005738Zero Blank G 062600B 6/26/00 ND ND na 0005738 Zero Blank H 0626OOB 6/26/00 ND m 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 ZeroBlank D 062700A 6/27/00 ND ND na 0005738Blank A 062800A 6/28/00 m ND na 0005738 Blank B 062SOOA malm ND ND na 0005735ZeroBlank C 062800A 6/28/00 ND ND na 0005738 Zero Blank D 062800A WKl ND ND na 0005738Blank A 0629ooA 6/29/00 0.0301 0.0262 na 0005738Blank B 062900A 6129/00 ND ND na 0005738 Zero Blank C 062900A 6/29/00 ND ND na 0005738 Zero Blank D 062900A 6/29mO ND ND na 0005738Blank A 063000A 6/30/00 - - ND ND na 0005738Blank B 063000A m m - N D ND na 0005738 Zero Blank C 063QOOA mol00 ND ND na 0005738ZeroBlankD 063000A 6/3o/oo ND . ND AVERAGE: c 0.0100 < 0.00869 STANDARDDEVIATION NQ NQ Note: The averageand standard deviation were calculatedby using 0.0100as the value ' for those samples reported as 0.0100,0.00869 forthose 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-U284--9--.. ! Centre Study No.: 023-016 SponsorProtocoi No.: FACT-TOX-110 I Table II. Summary of PFOS residues in G1-Control-GDO Sponsor Centre ID I Set Corrected Date Residue Found Residue Found 1950lF,FO,Gl-Control-GDO 0003977 19502F,FO,GlControl-GDO 0003978 19503F,FO,Gl-Control-GDO 0003979 19504F,FO,Gl-Control-GDO 0003980 19505F,FO,Gl-Control-GDO 0003981 19506~P0,G1-Control-GDO 0003982 . 19507F,FO,G1-Control-GDO0003983 19508F,FO,G1-Control-GDO 0003984 19509F,FO,G1-Control-GDO 0003985 1951OF,FO,Gl-Control-GDO 0003986 Number 061200AR 061200AR 061200AR 061200AR 061200AR 061200AR 061200AR 061200AR 061200AR 061200AR Extracted 6/12/00 6/12/00 6/12/00 6/12/00 6/12/00 6/12/00 6/12/00 6/12/00 6/12/00 6/12/00 . fHdg) ND ND < 0.0100 ND ND 0.0301 ND < 0.0100 0.0400 ND (P?d ND ND < 0.00869 ND ND 0.0262 ND 0.00869 0.0348 ND ' 19511F.FO,Gl-Control-GDO 0003987 061200AR 6l12100 ND ND 19512F,FO,Gl-Control-GDO 0003985 061200AR 6/12/00 ND ND 19513F,FO,Gl-ContrOl-GDO 0003989 061200AR 6/12/00 ND NR 19514F,FO.Gl-Control-GDO 0003990 061200AR 6/12/00 ND ND 19515F,FO,G1-Control-GDO 0003991 061200AR 6/12/00 ND m 19516F,FO.Gl-Control-GDO 0003992 06120OAR 6/12/00 0.0609 0.0529 AVERAGE: <0.0100 < 0.00569 STANDARDDEVIATION. NQ NQ ND = Not Detected and NQ = Not Quantifiable Note: The avenge and standard deviationwere calculatedby using 0,0100as the value for those samples reported as < 0.0100,0.00869 for those reported as < 0.00869, and zero for those reported as N D or NQ. 3M EnvironmentalLaboratory 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 i SponsorProtokol No.:FACT-TOX-110 j Table In. Summary of PFOS residues in G2-0.1MKD on GDO Sponsor CentrelD Set Corrected ' , Date Residue Found Residue Found ID Number Extracted (pg/g) (P gig) 1951~,FO,G2-0.lMJS.D-GD00O03993 061400AD 6/14/00 0.701 0.609 19518F,FO,G2-0.lMKD-GDO 0003994 061400A 6/14/00 0.456 0.396 19519F,FO,G2-0.lMKD-GDO 0003995 061400A 6/14/00 0.390 0.339 19520F,FO,G2-0.1MKD-GDO 0003996 061400A.D 6/14/00 0.676 0.587 1952lF,FO,G2-0,TMKD-GW 0003997 061400A 6/14/00 0.495 0.430 19522F,FO,G2-0.IMKD-GDO 0003998 061400A 6/14/00 0.381 0.331 19523F,FO,G2-0.1MKD-GW 0003999 061400A 6/14/00 0.458 0.398 19524F,FO,G2-O.lMKD-GDO 0004000 061400AD 6/14/00 0.833 ' 0.724 19525F,FO,G2-0.1MKD-GDO 0004001 061400A 6/14/00 . 0.473 0.41 1 19526F,FO,G2-0.1MKD-GDO OOO4002 061400A 6/14/00 0.446 0.388 19527F,FO,G2-0.IMKD-GDO 0004003 061400A 6/14/00 0.497 0.432 19528F,FO,G2-0.1MKD-GDO 0004004 061400AD 6/14/00 0.701 0.609 19529F,FO,G2-0.lMXD-GDO 0004005 0614OOAD 6/14/00 0.797 0.693 19530F,FO,G2-0.1~-GW0004006 061400A 6/14/00 0.394 0.342 19531F,FO,G2-O.lMKD-GDO 0004007 061400AD 6/14/00 0.735 0.638 19532F,FO,G2-0.1MKD-GW 0004008 061400AD 6/14/00 0.748 0.650 AVERAGE: 0.574 0.499 STANDARD DEVTATION: 0.160 0.139 Amended Page 23 of 103 . Page 358 1y 3M Medical Department Study: T-6295.12 I Analytical Report: FACT TOX-110 LRN-lJ2W- - .. _..- -. _.- Centre StudyNo.:023-016 ! Sponsor Protocol No.: FACT-TOX-110 1 Table Iv. Summary of PFOS residues in G3-0.4MKD on GDO I Sponsor ID CentreID I 19533F,FO,G3-0.4MXD-GDO ooo4009 19534F,FO,G3-0.4MKD-GDO 0004010 19535F,FO,G3-0.4MKD-GDO OOO4011 19536F,FO,G3-0.4MKD-GDO 0004012 19537F,FO,G3-0.4MKD-GDO OOO4013 19538F,FO,G3-0.4MKD-GDO 0004014 19539F,FO,G3-0.4MK.D-GDO ooo4015 19540F,FO,G3-0.4MKD-GDO 0004016 19541F,FO,G3-0.4MKD-GDO 0004017 19542F,FO,G3-0.4MXD-GDO oo04018 19543F,FO,G3-0.4MKD-GDO oo04019 19544F,FO,G3-0.4MKD-GDO 0004020 19545F,FO,G3-0.4MKD-GDO O004021 19546F,FO,G3-0:4MKD-GDO 0004022 19547F,FO,G3-0.4MKD-GW 0004023 Set Number 061500AD 061500AD 061500AD 061500AD 06 1500AD 061500AD 061500AD 061500AD 061500AD 06 1500AD 0615ObAD 061500AD 061600AD 0615OOAD 0615WAD Date Residue Found Extracted (pdg) 6/15/00 2.60 6/15/00 3.75 6/15/00 2.80 6/15/00 3.17 6/15/00 2.78 6/15/00 3.08 . 6/15/00 3.68 6/15/00 2.04 6/15/00 . 2.10 6/15/00 3.29 6/15/00 2.11 6/15/00 3.43 6116/00 2.76 6/15/00 2.33 6/15/00 2.22 Corrected Residue Found Wg) 2.26 3.26 2.43 2.76 2.41 2.68 3.20 1.78 1.83 2.86 . 1.84 2.98 2.39 2.03 1.93 19548F,FO.G3-0.4MKD-GW 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 L R. N--.U_-B-4._ 9__^-_ Centre Study No.:023-016 I SponsorProtocolNo.:FACT-TOX-110 Table V. Summary of PFOS residues in G4-1.6iMKD on GDO Sponsor , CentreID Set Date Residue Found ID Number Extracted 19549F,FO,G4-1.6MKD-GDO 0004025 061600AD 6/16/00 (pdg) 14.1 1955OF,FO,G4-1.6MKD-GDO 0004026 061600A.D 6/16/00 16.9 19551F,FO,G4-1.6MKD-GDO 0004027 061600AD 6/16/00 10.4 19552F,FO,G4-2.6MKD-GDO 0004028 061600AD 6/16/00 13,l 19553FS;O.G4-1.6MKD-GDO 0004029 0616OOAD 6/16/00 13.7 19554F,FO,G4-1.6MKD-GDO 0004030 061600AD 6/16/00 15.5 19555F.FO,G4-1.6MKD-GW 0004031 0616OoAD 6/16/00 10.3 19556F,FO,G4-1.6MKD-GDO 0004032 061600AD 6/16/00. 10.3 19557J?,FO,G4-1.6MKD-GDO0004033 061600A.D 6/16/00 8.96 19558F,FO,G4-1.6MKO-GDO 0004034 061600AD 6/16/00 6.85 19559F~O,G4-1.6MKD-GDO0004035 061600AD 6/16/00 7.68 19560F,FO,G4-1.6MXD-GDO0004036 061600AD 6/16/00 16.5 19561F,FO,G4-1.6MKD-GDO 0004037 0616OOAD 6/16/00 12.1 19562F,FO,G4-1.6MKD-GDO 0004038 061600AD 6/16/00 .9.59 19563F,FO.G4-1.6MKD-GDO 0004039 061600AD 6/16/00 7.47 - 19564F,FO.G4-I.~-GDO 0004040 061600AD 6/16/00 17.0 AVERAGE: 11.9 STANDARD DEVIATION: 3.45 Corrected Residue Found (!ME) 12.3 14.7 9.03 11.4 11.9 13.5 8.98 8.91 7.79 5.95 6.67 14.4 10.5 8.33 6.49 14.7 10.3 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 SponsorProtocol No.:FAV-TOX-110 Table VI. Summary of PFOS residues in G5-3.2MKD on GDO Sponsor CentreID Set Corrected Date Residue Found Residue Found ID 19565F,FO,G5-3.2MKD-GDO 0004041 19566F,FO,G5-3.2MKD-GDO0004042 19567F.FO,G5-3.2MKD-GDO 0004043 Number 061900AD 0619OOAD 061900AD Extracted 6/19/00 6/19/00 6/19/00 (pglg) ' 23.5 29.5 30.3 wg> 20.4 25.7 26.3 19568F,FO,G5-3.2MKD-GDO 0004044 061900A.D 6/19/00 24.2 21.0 19569F.FO,G5-3.2MKD-GDO 0004045 061900AD 6/19/00 27.1 23.5 1957OF,FO,G5-3.2MJCD-GDO0004046 061900AD 6/19/00 19.7 17.1 19571F,FO,G5-3.2MKD-GDO 0004047 061900AD 6/19/00 29.9 26.0 19572F.FO,Gs-3.2MXDsrW-GDO 0004045 061900AD 6/19/00 25.4 22.1 19573F,FO,G5-3.2MKD-GDO 0004049 061900AD 6/19/00 23.0 20.0 19574F,FO,G5-3.2I&.D-GDO 0004050 0619OOAD 6/19/00 29.3 25.5 19575F,FO,G5-3.2MKD-GDO0004051 061900AD 6/19/00 23.0 20.0 19576F,FO,G5-3.2MKD-GDO 0004052 061900AD 6/19/00 36.3 31.6 19577F,FO,G5-3.2MKD-GDO 0004053 061900AD 6/19/00 34.2 29.8 19578F,.FO,G5-3.2MKD-GDO0004054 061900AD 6/19/00 . 34.8 30.2 19579F,FO,G5-3.2MKD-GDOQ004055 061900AD 6/19/00 25.4 22.0 1958OF,FO,G5-3.2MxD-GDO 0004056 0619OOA.D 6/19/00 24.9 21.6 AVERAGE: 275 239 STANDARD DEVIATION 4.7s 4.15 3M Environmental Laboratory Amended Page 26 of 103' Page 361 3M Medical Department Study: T-6295.12 I Analytical Report: FACT TOX-110 LRN-U2849 ... . . .. 0: 'I CentreStud'y No.:023-016 SponsorProtocol No.:FACT-TOX-I 10 I Table VI.Summary of PFOS residues in G1-Controlon GD7 Sponsor CentreID Set Corrected Date ResidueFowd ResidueFound IID 19501F,FO,GI-Control-GD7 0004057 1950?F,FO,G1-Control-GD70004058 19504F,FO.Glontrol-GD7 0004059 19505F,FO,G1-Control-GD7 0004060 Number 062000A 062000A 062000A 062000A Extracted 6/20/00 6/20/00 6/20/00 6/20/00 {pdg) ND NQ NQ ND hh!) ND NQ NQ ND 19506F,FO,Gl-Control-GD70004061 062000A 6/20/00 0,0178 0.0155 19507F,FO,G1ontrol-GD7 OOO4062 19508F,FO,G1-Control-GD7'0004063 l95O9F,FO,Glontrol-GD7 0004064 19510F,FO,Gl-ControI-GD7 0004065 1951 lF,FO,G1-Control-GD7 . 0004066 19512F,FO.Gl-ControLGD7 0004067 19513F,FO,Gl-Control-GD70004068 19514F,FO,Gl-Control-GD7 0004069 19515F,FO,Gl-Control-GD7 0004070 19516F;FO,Gl-Control-GD7-0004071 062000A 6/2WOO 062000A 6/20/00 062000A 6/20/00 062000A 6/20/00 0620oOAR 6/20/00 062000AR 6/20/00 062000A 6/20/00 062000A 6/20/00 062000AR 6/20/00 062000AR 6/20/00 AVERAGE: ND c 0.0100 . ND ND Nu ND ND ND ND NQ 0.0100 ND < 0.00869 ND ND ND ND ND ND ND NQ < 0.00869 S T W A R D DEVIATION NQ NQ ND =Not Detected andNQ = Not Quantifiable Note: The average and standarddeviationwere calculatedby 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 i Analytical Report: FACT TOX-110 . LRN-U2849- ! Cenue Study No.:023-016 . t Sponsor Protocol No.: FACT-TOX-110 'Tab&wn. Summary of PFOS residues in G2-0.1MK.Don GD7 1 Sponsor CentreID Set Corrected Date Residue Found Residue Found ID Number Extracted 19517F,FO,G2-0.lMKD-GD7 0004072 062100A 6/21/00 (pdp) 0.409 (Pel 0.356 19518F,FO,G2-0.1MKD-GD7 0004073 062100AD 6/21/00 0.56 1 0.488 19519F,FO,G2-0.IMKD-GD7 0004074 062100A 6/21/00 0.345 0.299 19521F,FO,G2-0.1MXD-GD7 0004075 062100A 6/21/00 0.465 0.404 19522FzO,G2-0.1MJCD-GD7 0004076 062100AD 6/21/00 0.778 0.676 19523F,FO,G2-0.lMKD-GD7 0004077 062100AD 6/21/00 0.625 0.543 19524F,FO,G2-0.1MKD-GD7 0004078 062100AD 6/21/00 0.576 . 19525F,FO,G2-0.1MKD-GD7 0004079 062100AD 6/21/00 . 0.595 0.500 0.5 17 19526F,FO,G2-0.1MKD-GD7 0004080 062100A 6/21/00 0.469 0.408 19527F,FO,G2-0.1MKD-GD7 0004081 062100AD 6/21/00 0.742 0.645 19528F,FO.G2-0.1MKD-GD7 0004082 062100A.D 6/21/00 0.532 0.462 19529F,FO,G2-0.1MJSD-GD70004083 062100AD 6/21/00 0.744 0.647 19530F,FO,G2-0.1MKD-GD7 0004084 062100A 6/21/00 0.424 0.369 1953lF,FO,G2-0.lMKD-GD7 0004085 062100AD 6/21/00 19532F.FO,G2-O.liMKD-GD7 0004086 062100A.D 6/21/00 AVERAGE: STANDARD DEVIATION 0.624 0.574 0.5& 0.128 0.542 - 0.499 0.490 . 0.111 3M EnvironmentalLaboratory Amended Page 28 of 103 Page 363 3M Medical Department Study: T-6295.12 I Analytical Report: FACT TOX-110 - --- LRN-U2849 .__.____-.- ! 1 ! Centre Study No.:023-016 SponsoiProtocol XO.:FACT-TOX-110 Table M.Summary of PFOS residues in G3-0.4MKD on GD7 Sponsor CentreID Set Date Residue Found ID Number Extracted (pe/g) 19533F,FO.G3-0.4MKD-GD7 0004087 062200AD 6/22/00 2.55 19534F,FO,G3-0.4MKD-GD7 0004088 062200AD W00 2.69 19535F,FO,G3-0.4MKD-GD7 0004089 062200AD 6/22/00 1.82 19536F,FO,G3-0.4MKD-GD7 OOO4090 062200AD 6/22/00 3.26 19537F,FO,G3-0.4MKD-GD7 0004091 062200AD 6/22/00 2.61 19538FJO,G34,.4MKT)-GD7 0004092 0 6 2 2 0 6/22J00 2.93 19539F,FO,G3-0.4h4KD-GD70004093 062200AD 6/22/00 2.09 19540F,FO,G3-0.4MKD-GD7 0004094 062200AD 6/22/00 2.27 19542FJO,G3-0.4MKD-GD'7 0004095 062200AD 6/22/00 3.00 19543F,FO,G3-0.4i'vfKD-GD7 0004096 062200AD 6122/00. 2.80 . 19544F,FO,G3-0.4MKD-GD7 0004097 062200AD 6/2/00 2.28 1954@,FO,G3-0.4MKD-GD7 0004098 062200AD 6/22/00 1.52 19547F,FO,G3-0.4MKD-GD7 OOO4099 062200AD 6/22/00 2.06 - 19548F,FO,G3-0.4MKD-GD7 0004100 0622DoAD 6/22/00 2.85 AVERAGE: 2.45 STANDARD DEVIATION 0.192 Corrected Residue Found (P&) 2.21 2.34 1.59 2.83 2.27 2.55 . 1.82 1.97 2.61 2.43 1.98 1.32 1.79 2.47 2.16 0.428 .. Amended Page 29 of 103 Page 364 3M Medical DepartmentStudy: T-6295.12 I . .. . . . . Analytical Report: FACT TOX-110 .. LRN-U2849 ' ! ! Centre Study NO.: 023-016 Sponsor ProrocoI No.: FACT-TOX-110 Table X. Summary of PFOS residues in G4-1.6iMKD on GD7 Sponsor CentreID Set iDate Residue Found ID Number Extracted 19550F,FO,G4-1.6MKD-GD70004101 062200BD 6/22/00 (pe/g> 15.0 19553F,FO,G4-1.6MKD-GD7 o004102 062200BD 6/22/00 12.3 19554F,FO,G4-1.6MKD-GD7 0004103 062200BD 6/22/00 11.8 19555F,FO,G4-1.6MKD-GD7 OOO4104 062200BD 6/22/00 9.61 19556F,FO,G4-1.6MKD-GD7 0004105 062200BD 6/22/00 10.2 19558F,FO,G4-1.6MKD-GD7 0004106 062200BD 6122/00 5.89 . 19559F,FO;G4-1.6MKD-GD7 OOO4107 062200BD 6l22/00 6.89 1956OF,FO,G4-1.6MKD-GD7 ooo4108 06220033D 6/22/00 7.02 1956lFzO,G4-I.6MKD-GD7 0004109 0622CQBD 6/22/00 8.88 I9562F,EO,G4-I.6MKD-GD7 oO04110 062200BD 6/22/00 10.8 ' 19563F,FO,G4-1.6MKD-GD7 0004111 062200BD 6/22/00 14.9 19564F,FO,G4-1.m-GD70004112 06ZZMBD 6/Z@o 13.7 AVERAGE: 10.6 STANDARD DEVIATION: 3.08 Corrected Residue Found (Pa) 13.0 10.7 10.2 8.35 8.88 5.12 5.99 6.10 . 7.71 9.42 13.0 11.9 9.19 2.67 3M EnvironmentalLaboratory Amended Page 30 of 103 Page 365 3M Medical Department Study: T-629. 5.1.2 ! Analytical Report: FACT TOX-110 C R.-N. 4_2_8-4-9_-_- Centre StudyNo.:023-016 ! Sponsor Protocol No.:FACT-TOX-110 Table XI. Summary of PFOS residues in G5-3.2PIKD on GD7 Corrected i Sponsor Centrem Set Date Residue Found Residue Found m Number Exacted (p&) (Id.& 19565F,FO,G5-3.2MKD-GD70004113 062300AD 6/23/00 51.7 44.9 19566F,FO,G5-3.2MKDGD7 0004114 0623OOAD 6/23/00 26.5 23.0 19567F,FO,G5-3.2MKD-GD7 0004115 062300AD 6/23/00 35.9 31.2 19568F,FO,G5-3.2MKD-GD70004116 062300AD 6/23/00 23.0 20.0 19569F.FO.GS-3.2MKD-GD7 0004117 062300AD 6/23/00 42.9 37.3 1957OF,FO,G5-3.2MJSD-GD70004118 062300AD 6/23/00 37.4 32.5 1957lF,FO,G5-3.2MICD-GD70004119 062300AD 6/23/00 43.0 37.4 19572F,FO,G5-3.2MKD-GD7 0004120 0623'MAD 6/23/00 38.8 33.7 19573F,FO,G5-3.2MKD-G070004121 062300AD 6/23/00 26.7 23.2 19574F,FO,G5-3.2MKD-GD7 0004122 0623MAD 6/23/00 36.3 31.6 19575F,FO,G5-3.2MI(D-GD07004123 062300AD 6/23/00 35.5 30.8 19577F,FO,G5-3.2MKD-GD70004124 062300AD 6/23/00 68.3 59.4 19579F,FO,G5-3.2MKD-GD7 0004125 062300AD 6/23/00 35.9 31.2 1958OF,FO,G5-3.~-GD70004126 062300AD 6/23;100 30.1 26.1 AVERAGE. 38.0 33.0 STANDARDDEVIATION: 11.5 10.0 3M Environmental Laboratory Amended Page 3 1 of 103 Page 366 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-mn84-9.--. * ! I TableXII. Summary of PFOS &dues Centre Study No.:023-016 SponsorProtocol No.:FACT-TOX-110 in G1-Control on GD15 Sponsor CentreID, Set Date Residue Found m Number Extracted 1950lF,J?O,G1-Control-GD15 0004127 062300B 6i23/00 (pdg) ND 19502F,FO,Gl-Control-GDtS 0004128 062300B 6/23/00 ND l95O4F,FO,Gl-Control-GD15 0004129 062300B 6/23/00 ND 1950SF,FO,Gl-Control-GD150004130 0623OOB 6/23/00 "D 19506F,FO,G1-Control-GD15 0004131 062300B 6/23/00 0.0445 1950~~O,G1-Control-GD150004132 062300B 6/23/00 ND 19508F,FO,G1-ConwLGD-GD15 0004133 0623008 6/23/00 ND 19509F,FO,G1-Control&Dl5 0004134 0623OOB 6/23/00 0.0242 1951OF~O,Gl-ControlGD150004135 0623OOB 6/23/00 <0.0100 1951~F,FO,GI-COU~~~I0-0G04D13~6~ 062300B 6/23/00 ND 19512FJ?O,G1-Control-GDl50004137 062300B 6/23/00 0.0579 19513F,FO,G1-ControI-GDl5 0004138 062300B 6/23/00 ND 19514F~O,Gl-Control-GD15 0004139 062300B 6/23/00 ND 1951SF,FO,Gl-ControlGD150004140 062300B 6/23/00 ND 19516F,FO,G1-Control-GD15 0004141 062300B 6/23/w 0.0216 AVERAGE: 0.0105 STANDARD DEVIATION: 0.0185 Corrected Residue Found (Pdd ND ND ND ND 0.0387 ND ND 0.0210 < 0.00869 ND 0.0503 m ND ND 0.0188 0.00917 0.0161 ND = Not Detected and NQ = Not Quantifiable Note: ne average and standard deviation were calculated by using 0.0100 as the value for those samples reported as <0.0100,0.00869for those reportedas < 0.00869, and zero for those reportedas ND or NQ. 3M EnvironmentalLaboratory Amended Page 32 of 103 Page 361 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 .. ._-L...R.N-.----U-2849 ! Centre Study No.: 023-016 I SponsorProtocol No.:FACT-TOX-110 ! Table Xnl. Summary of PFOS residues in G2-0.1MKD on GD15 Sponsor C e n m p Set Corrected Date Residue Found Residue Found ID 195 17F,FO,G2-0.lMKD-GD15 0004142 19518F,FO,G2-0.1MKD-GD150004143 19519F,FO,G2-O.1MKD-GDI50004144 19521F,FO,G2-0.1MKD-GDIS0004145 19522F,FO,G2-0.1MKD-GD150004146 19523FJ?O,G2-O.lMKD-GD150004147 19524F,FO,G2-0.lMJSD-GD150004148 19525F,FO,G2-O.lMxD-GD1.05004149 19526F,FO,G2-0.IMKDGDlS 0004150 19527F,FO.G2-O.IMKD-GDloSoO4151 19528F,FO,G2-0.1MKD-GD150004152 19529F,FO,G2-0.1MKD-GD150004153 19530FJ?O,G2-0.IMKD-GDIS 0004154 Number 062600AD 0626OoAD 062600AD 062600AD 0626OoAD 062600AD 062600AD 062600AD 0626M1AD 062600AD 062600AD 062WAD 062600AD Extracted 6/26/00 6/26/00 6/26/00 6/26/00 6/26/00 6/2goO 6/26/00. 6/26/00 6/26/00 6/26/00 6/26/00 6/26/00 6/26/00 (pdg) 0.758 0.672 0.782 0.632 0.817 0.642 0.834 1.08 . 0.778 0.799 0.845 0.724 0.724 (PglP) 0.659 0.584 0.680 0.549 0.710 0.558 0.725 0.939 0.676 0.694 0.734 0.629 0.630 1953IF.FO,G2-0.IMKD-GDI5 0004155 062600AD 6/26/00 19532F.FO.G2-0.IMKD-GDlS 0004156 062600Ai) 6/26/00 AVERAGE: STANDARDDEVLk'ION: . 0.777 0.568 0.762 0.119 0.675 0.434 0.662 0.104 3M Environmental Laboratory Amended Page 33 of 103 Page 368 3M Medical Department Study: T-6295.12 I - Analytical Report: FACT TOX-I 10 - . ._____tR_N-_U2849 ! . Centre StudyNO.:023-016 SponsorProtocol No.:FACT-TOx-110 Table XIV. Summary of PFOS residues in G3-0.4MKD on GD15 1 Sponsor CentreID Set Corrected Date Residue Found Residue Fpund ID 19533F,FO,G3-0.4MKD-GDlS 0004157 19534F,FO,G3-0.4MKD-GDl50004158 19535F,FO,G3-0.4MKD-GDl05004159 19536F,FO,G3-0.4MKD-GD150004160 19537F,FO,G3-0.4MKD-GD150004161 19538F.FO,G3-0.4MKD-GD150004162 19539F,FO,G3-0.4MKD-GD150004163 19540F,FO,G3-0.4MXD-GDl50004164 Number 062600BD 062600BD 062600BD 062600BD 062600BD 062600BD 062600BD 062600BD Extracted 6/26/00 6/26/00 6/26/00 6/26/00 6/26/00 6/26/00 6/26/00 6/26/00 (pgg) 2.76 4.82 3.63 3.19 3.40 3.14 3.09 4.15 (CLdP) 2.40 4.19 3.15 2.77 2.95 2.73 2.69 3.61 19542F,FO,G3-0.4h4KD-GD15OW165 062600BD 6/26/00 1.88 1.63 19543F,FO,G3-0.4hfKD-GDlS0004166 062600BD 6/26/00 3.42 2.97 19544F,FO,G3-0.4MKD-GDIS0004167 062600BD 6/26/00 3.37 2.93 19546F,FO,G3-0.4MKD-GD150004168 062600BD 6/26/00 2.63 2.28 19547FJ?O,G3-0.4MKD-GD105004169 062600BD 6/26/00 3.86 3.36 19548F,FO.G3-0.4MXD-GD150004170 062600BD 6/26/00 3.91 . 3.40 AVERAGE: 338 STANDARD DEVIATION: 0.713 2.93 0.620 3M Environmental Laboratory Amended Page 34 of 103 Page 369 3M Medical Department Study: T-6295.12 I. Analytical Report: FACT TOX-110 .LRN~Ct.284-4- '! Centre Study No.:023-016 SponsorProtocol No.:FACT-TOX-110 Table XV. S&ry , of PFOS residues in G4-1.6hIKD on GD15 , Sponsor CentreID Set Corrected Date Residue Found Residue Found ID Number Extracted (&E) 1955OF,FO,G4-1.6MKD-GD150004171 062700AD 6/27/00 13.9 (PdE9 12.1 19553F,FO.G4-1.6MKD-GDl50004172 062700AD 6/27/00 14.8 12.9 19554F,FU.G4-1.6MKD-GD15 OOO4173 062700AD 6/27/00 14.2 12.3 19555F,FO,G4-1.6MKD-GD150004174 062700AD 6/27/00 16.3 14.1 19556F,~O,G14.-6MKD-GD15 ooo4175 062700AD 6/27/00 9.07 7.88 19558F,FO,G4-1.6MKD-GDl50004176 062700AD 6/27/00 7.17 6.23 19559F,FO,G4-1.6MKD-GDl50004177 062700AD 6/27/00 7.48 6.50 1956OFJO,G4-1.6MKD-GD15 0004178 062700AD 6/27/00 14.9 ' 12.9 19561F,FO,G4-1.6MKD-G1D5 0004179 062700AD 6/27/CQ 17.4 15.2 19562F,FO,G4-1.6MKD-GDl50004180 . 062700AD 6/27/00 11.4 9.92 19563F,FO,G4-1,6MKD-GD105004181 062700AD 6/27/00 8.95 7.78 19564F,FQ,G4-1.6vII(D-c;D150004182 062700AD 6/27/00 17.3 15.1 AVERAGE: 12.7 11.1 STANDARDDEVIATION 3.77 3.28 .. 3M tnvironmental LaDoraroV Amended Page 35 of 103 ..- Page 370 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 _tR_N-.-.--U--2_8- 49 ! Centre Study No.:023-016 Sponsor Protocol No.: FACT-TOX-110 Table XVI. Spmmary of PFOS residues in G5-3.2MKD on GD15 sponsar CentreID Set Corrected Date Residue Found ResidueFound ID Number Extracted 19565F,FO,G5-3.2MKD-GDl50004183 062800AD 6/28/00 (p&) 35.2 (Pd/lr) 30.6 19566F,FO,G5-3.2MKD-GD15 0004184 062800AD 6/28/00 15.2 13.2 19567F,FO.G5-3.2MKD-GDl50004185 062800AD 6/28/00 20.4 17.7 19.568F,FO,G5-3.2MKD-GDlS0004186 062800AD 6/28/00 46.9 40.8 19569F,FO,G~3.2MKD-GDl0S004187 062800AD 6/28/00 36.3 31.6 19570F,FO,G5-3.2MKD-GDl50004188 062800AD 6/28/00 31.1 27.0 19571FzO,G5-3.2i?lKD-GDl50004189 062800AD 6/28/00 28.4 24.7 1957ZF,FO,G5-3.2MKD-GD15OOO4190 062800AD 6/28/00 20.5 17.9 19573F,FO,G5-3.2MKD-G1D5 OOO4191 062800AD 6/28/00 39.8 34.6 19574F,FO.GS-3.2MKD-GDlS0004192 062800AD 6/28/00 47.3 41.1 1957SF,FO,GS-3.2MKD-GDl50004193 062800AD 6/28/00 40.7 35.4 19577F,FO.G5-3.2MKD-GDl50004194 062800AD 6/28/00 47.3 41.1 19579F,FO0,G5-3.2MKD-GD105004195 062800AD 6/28/00 33.4 29.1 19580F,FO,G5-3.2MKD-GD15 0004196 062800AD 6/28/00 32.7 AVERAGE 339 STANDARD DEVIATION 10% 28.4 29.5 - 831 3M Environmental Laboratory Amended Page 36 of 103 Page 371 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 - LRN_-U_2_8._4,9....- 1.. CentrelStudy No.: 023-016 SponsorProtocol No.:FACT-TOX-110 Table m.Summary of PFOS residues in G1-Control'onGD21 Sponsor CentreID Set Corrected Date Residue Found Residue Found ID19504F,FO,Gl-Control-GD210004197 19505FJ?O,G1-Control-GD21 004198 Number 062900A 062900A Extracted 6/29/00 6/29/00 (pe/g) ND ND (P_g/d ND ND 19506F,FO,Gl-Control-GD210004199 O O o A 6/29/00 19507F.FO,Gl-Control-GD210004200 062900A 6/29/00 19509F,FO,G1-Control-GD210004201 0629OOA 6/29/00 19512F,FO,Gl-Control-GD210004202 0629oOA 6/29/00 19516F~,Gl-Contml-GD21 0004203 062900A 6/29/00 AVERAGE: STANDARD DEVIATION. 0.0123 ND 0.0843 0.0215 < 0.0100 0.0183 0.0302 0.0107 ND 0.0733 0.0187 c 0.00869 0.0159 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 samplesreported as <0.0100,0.00869for those reported as < 0.00869, and zero . for those reported as ND or NQ. Table XVm. Summary of PFOS residues in G2-0.1MKI) on GD21 Corrected Sponsor CentreID Set Date Residue Found Residue Found m Number Extracted (p&) (Pdd 19518F,FO,G2-O.IMKD-GD210004204 0629-3ooOA.D 6/29/00 . 0.735 0.639 195 19F,FO,G2-O.lMKD-GD21 0004205 062900A . 6/29/00 0.473 0.411 19521F,Ffl,G2-0.1MKD-GD21 0004206 . 0629OoA 6/29/00 0.403 0.351 19523F,.FO,G24.1MKD-GD201004207 062900A 6/29/00 0.402 0.349 19524F,.FO,G2-0.1MKf)-GD210004208 062900A 6/29/00 19528F,FO,G2-O.lMKD-GD21 0004209 062900A 6/29/oq 0.494 0.456 0.429 0.396 -. 19529F.FO.GZ-o. IMKDGD21 0004210 0629OOA 6/29/00 AVEIUGE STANDARD DEVIATION: 0.39 1 0.479 .0.120 0.340 0.417 0.104 3M Environmental Laboratory Amended Page 37'of 103 Page 372 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LR.N..-.U-.2-.8--49 - ! CentreStudy No.:023-016, ! SponsorProtocol No.:FACT-TOX-110 Table XIX. Summary of PFOS residues in G3-0.4- on GD21 Sponsor CentreID Set Corrected Date Residue Found Residue Found ID Number Extracted 19533F,FO,G3-0.4MKD-GD210004211 0629-3OOOAD 6/30/00 19534F,FO,G3-0.4MKD-GD210004212 0629-3OOOAD 6/30/00 19536F,FO,G3-0.4MKD-GD210004213 0629-3OOOAD 6/30/00 19543F,FO,G3-0.4MKD-GD210004214 0629-3000AD 6/30/00 19546F,FO,G3-0.4MKD-GD210004215 0629-3ooOAD 6/30/00 19548F,FO.G3-0.4MKD-GDZl0004216 0629-3ooOAD 6/30/00 AVERAGE: STANDARD DEVIATION: (pdg) 1.68 3.10 1.85 2.23 2.u 5.42 2.75 139 (P.m 1.46 2.70 1.61 1.94 1.94 4.71. 239 1.21 TabIe XX. S h a r y of PFOS residues in G4-1.6MKD on GD21 Sponsor CentreID Set Date Residue Found ID Number Extracted (pg/g) 19556F,FO,G4l.&-GDZ 1 0004217 0629-3OOOAD 6/30/00 18.3 19559F.FO,G41.6MKD-GD21 0004218 0629-3OOOAD 6/30/00 10.6 1956OF,FO,G4-1.6MKD-GD210004219 0629-3ooOA.D 6/30/00 11.2 19562F,FO,G4-1.6MKD-GDZlooo422O 0629-3OOOAp 6/3O/W 5.68 AVERAGE: 11.4 STANDARD DEVIATION: 5.19 Corrected Residue Found wid 15.9 9.19 9.72 4.94 9.93 4.51 Table XXI. Summary of PFOS residues in G5-3.2MKD on GD21 Sponsor CentreID Set Date Residue Found ID Number Extracted (p&) 19568F,F(J,G5-3.2MKD-GD2o1OO4221 0629-3ooOAD 6/3o/oo 25.1 19.569F,FO,G5-3.2MKD-GD210004222 0629-3oooAD 6nQ100 21.7 1957lF,FO,G5-3.2MKD-GD21 0004223 0629-3OOOAD 6/30/00 21.5 19572F,FO,G5-3.2IkD-GD210004224 0629-3ooOAD 6/30/00 15.0 19577F,FO,G5-3.2MKD-GD210004225 0629-3OGQA.D 6/30Mo 27.1 19579F,FO,G5-3.2MKD-GD210004226 0629-3000AD 6nQ/OO 28.3 AVERAGE: 23.1 STANDARD DEVIATION: 4.53 Corrected Residue Found (Pg/fz) 21.8 18.8 18.7 13.0 23.5 24.6 20.1 4.20 Amended Page 38 of 103 Page 373 v-3 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: FACT TOX-110 LRN-U2849 - 1CEntrG Analytical laboratorks, 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/Z/OO) Centre Analytical Laboratories COA Reference #: 023-018B Test Name Purity' Appearance Identification NMR Metals (ICPMS) 1. Calcium 2. Magnesium 3. Sodium 4. Potassium' 5. Nickel 6. Iron 7. Manganese Total Yo Irnburitv (-NIvlR) rotal% Impurity [LCMS) Total % Impurity (GCMS) Related Compounds POAA Residual Solvents (TGA) Purity by DSC Inorganic Anions (IC) 1. Chloride 2. Fluoride 3. Bromide 4. Nitrate 5 . Nitrite 6. Phosphate 7. Sulfate' Organic Acids ' (IC) 1. TFA 2. PFPA 3. HFBA 4. NFPA Elemental Analysis': 1. Carbon 2. Hydrogen 3. Nitrogen 4. Sulfur 5. Fluorine COA023-0 18B 3M Environmental Laboratory Specifications White Crvstalline Powder Result Conforms Positive 1. 0.017 wtJwt.% 2. 0.007 wt./wt.% 3. 1.355 wt./wt.% 4. 6.552 wtJwt.% 5. 0.003 wt.lwt.% 6 . 0.004 wtJwt.Yo 7. <0.001 wt./wt.% I 1.oo wt./wt.% I 10.60 wt./wt.% None Detected Theoretical Value = 0% Theoretical Value = 5.95% Theoretical Value = 60% 0.30 wt./wt.% None Detected Not A ~ d i c a b l e ~ 1. <0.015wt./wt.% 2. 0.27 wt./wt.% 3. <0.040wt./wt.% 4. <0.009 wt./wt.% 5 . (0.006 wt./wt.% 6. ~0.007wt./wt.% 7. 8.82 wt./wt.% 1. <0.1 wt./wt.% 2. <0.1 wt./wt.% 3. <0.1 wt./wt.% 4. <0.25 wt./wt.% 1. 12.08 wtJwt.% 2. 0.794 wtJwt.% 3. 1.61wtJwt.% 4. 10.1 wt./wt.% 5. 50.4 wt./wt.% Page 1 of3 Page 375 3 M Med- ical Department Study: T-6295.12 Analytical Report: FACT TOX-110 a\ 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 Centre Analytical Laboratories COA Reference #: 023-018B Date of Last Analysis: 0813 1/00 Expiration Date: 08131/01 Storage Conditions: Frozen I-10C Re-assessment Date: 08131/01 'Purity = 100%- (sum of metal impu,,cs, 1.39% +LC/MS impuI es, 10.60%+InorganicFluoride, 0.27%+NMR impurities, 1.OO%+ POAA, 0.30%) Total impurity fiom all tests = 13.56% Purity = 100% - 13.56% = 86.4% 'Potassium is expected in this salt form and is therefore not considered an impurity. 'Purity by DSC is generally not applicable to materials of low purity. No endotherm was observed for this sample. 4Sulfurin 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. HFBA NFPA PFPA Trifluoroacetic acid Heptafluorobutyric acid Nonofluoropentanoic acid Pcntafluoropropanoic acid 6Theoreticalvalue calculationsbased on the empirical formula, C~FI,SOJX+(MW=538) This work was conducted under EPA Good Laboratory Practice Standards (40 CFR 160). COA023-018B 3 M Environmental Laboratory Page 2 of 3 Page 376 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 a\ LKN-U284Y C m t w 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 #: 023-018B LC/MS Purity Profile: Impurity c c4s C6 c7 Total WtJwt. Yo 1.56 6.38 1.63 10.60 Note: The C4 and C6 values were calculatedusing the C4 and C6 standard calibration curves, respectively. The C5 value was calculated using the average response factors fromthe C4 and C6 standard curves. Likewise, the C7 value was calculated using the average response factors fiom the C6 and C8 standard curves. Prepared By: / u / ; z L = p ?A/- B $ d ~ ~ Reviewed Dawd S.Bell C&njyy'y'tkal Laboratories Date dh o h Flaherty Date Laboratory Manager, Centre Analytical Laboratories i COA023-018B Page 3 of 3 3M Environmental Laboratory Page 377 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 a\ LRN-U2849 CEntre Analytical Laboratorks. Inc. 3048 Research Drive State College,PA 16801 Phone: (814) 231-8032 Fax: (814) 231-1253 or (814) 231-1580 t INTERIM CERTIFICATE OF ANALYSIS Revision l(9/%/00) Centre Analytical Laboratories COA Reference #: 023-018A Test Name Purity' Appearance Identification NMR Metals (ICPiMS) I 1. calcium- 2. Magnesium 3. Sodium 4. Potassium* 5. Nickel 6. Iron 7. Manganese Total % Impurity (NMR) Total % Impurity (LCiMS) rota1 % ~ m. ~ u-r i t ~ (GWS) Related Compounds- POAA Residual Solvents (TGA) Purity by DSC Inorganic Anions (IC) 1. Chloride 2. Fluoride 3. Bromide 4. Nitrate 5. Nitrite 6. Phosphate 7. Sulfate` Organic Acids '(IC) 1. "FA 2. PFPA 3. HFBA 4. NFPA Elemental Analysis? 1. Carbon 2. Hydrogen 3. Nitrogen c 4. Sulfur 5. Fluorine COA023-018A 3M Environmental Laboratory Specifications White Crystalline Powder 1. Theoretical Value = 17.8% 2. Theoretical Value = 0% 3. Theoretical Value = 0% 4. Theoretical Value = 5.95% 5. Theoretical Value = 60% Result 86.9% Conforms Positive 1. 0.005 wt./wt.% 2. 0.001 wt./wt.% 3. 1.439 wt.lwt.% 4. 6.849 wt./wt.% 5* co.00 1wtJwt.% 6. 0.005 wt./wt.% 7. <0.001 wtJwt.% 1.93 wtJwt.% 8.41 wtJwt.% None Detected 0.33 wt./wt.% None Detected Not Auulicable' ~ ~~ 1. <0.015 \vt./wt.% 2. 0.59wt./wt.% 3. C0.040 wt./wt.% 4. <0.009 wt./wt.% 5. <0.006wt./wt.% 6. <0.007 wt./wt.% 7. 8.76 wt./wt.% 1. co.1 wt./wt.% 2. co.1 wt./wt.% 3. 0.10 wt./wt.% 4. 0.28 wt./wt.% 1. 12.48 wt./wt.% 2. 0.244 wt.lwt.% 3. 1.74 wt./wt.% 4. 8.84 wt./wt.% 5. 54.1 wt./wt.% Page 1 of 3 Page 378 3 M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LRN-U2849 atl!l!i\ Cmtre Analytical Laboratories, Inc. 3048 Research Drive Phone: (814) 231-8032 State College, PA 16801 Fax: (814) 231-1253 or (814) 231-1580 INTERlM CERTIFICATEOF ANALYSIS Centre Analytical Laboratories COA Reference #: 023-018A Date of Last Analysis: 0813 1/00 Expiration Date: 08/31/01 Storage Conditions: Frozen 5-10"C Re-assessment Date: 08/31/01 IPurity = 100%- (sum of metal impurities, 1.(5% -t C/MS impurities, 8.41%t-iorganic Fluoride, 0.59%+NMR impurities, 1.93%+0rganicacid impurities, 0.38%+POAA, 0.33%) - Total impurity fiom all tests = 13.09% PUrity = 100% 13.09% = 86.9% *Potassium is expected in this salt form and is therefore not considered an impurity. 3Purityby DSC is generally not applicableto materials of low purity. No endothermwas observed for this sample. 4Sulfurin 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 determinationin the elemental analysis, lending confidenceto this interpretation. Based on the results, the SO4 is not considered an impurity. 'TFA HFBA NFPA PFPA Trifluoroacetic acid Heptafluorobutyric acid Nonofluoropentanoic acid Pentafluoropropanoicacid 6Theoreticalvalue calculationsbased on the empirical formula, CsF17S03X' (MW=538) This work was conducted under EPA Good Laboratory Practice Standards (40 CFR 160). COA023-018A 3 M Environmental Laboratory Page 2 of 3 Page 379 3M Medical Department Study: T-6295.12 Analytical Report: FACT TOX-110 LKN-U284Y 3048 Research Drive Phone: (814) 231-8032 State College, PA issOr Fax: (814) 231-1253 or (814) 231-1580 INTERIM CERTIFICATE OF ANALYSIS Centre Analytical Laboratories COA Reference #: 023-018A LC/MS Purity Profile: Impurity I wt./wt. Yo c4 1.22 C6 4.72 I c7 1.14 Total I 8.41 1 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 firom 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: Laboratories Y//OfW Date o h Flaherty Date e a b o r a t o l y 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-I 10 LRN-U2849 Marvin T. Case, D.V.M., Ph.D., Study Director fite John L. Butenhoff, Ph.D., Sponsor Representative Date William Reagen, Ph.D., Laboratory Manager Kris 3. Hansen,-Ph.D.,Principle Analytical Investigator Date OS/O v/o/ Date 3M Environmental Laboratory Page 381