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3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Analytical Report: FACT TOX-001 LRN-U2103 Study Title 104-Week Dietary Carcinogenicity Study with Narrow Range (98.1%) N-Ethyl PerfluorooctanesulfonamidoEthanol in Rats Analytical Laboratory Report Title Determination of the Presence and Concentration of PFOS, PFOSA, PFOSAA, EtFOSE-OH, M556, and PFOSEA in Serum and Liver Samples of Crl:CD@(SD)IGS BR Rats Exposed to N-Ethyl PerfluorooctanesulfonamidoEthanol Data Requirement Not Applicable Author 3M Environmental Laboratory Study Completion Date JuAntes0ig6n,in2g001 Performing Laboratories Liver and Serum Analyses 3M Environmental Laboratory Building 2-3E-09, 935 Bush Avenue St. Paul, MN 55106 Project Identification 3M Medical Department Study: T-6316.1 Covance In-Life Study: 6329-212 Analytical Report: FACT TOX-001 3M Laboratory Request No. U2103 Total Number of Pages #3#1#0 3M Environmental Laboratory 3M Environmental Laboratory Page 1 Page 1 3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Analytical Report: FACT TOX-001 LRN-U2103 This page has been reserved for specific country requirements. 3M Environmental Laboratory 3M Environmental Laboratory Page 2 Page 2 3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 GLP Compliance Statement Analytical Report: FACT-TOX-001 LRN-U2103 Analytical Report: FACT TOX-001 LRN-U2103 Analytical Laboratory Report Title: Determination of the Presence and Concentration of PFOS, PFOSA, PFOSAA, EtFOSE-OH, M556, and PFOSEA in Serum and Liver Samples of Crl:CD@(SD)IGS BR Rats Exposed to N-Ethyl Perfluorooctanesulfonamido Ethanol Study Identification Numbers: T-6316.1, FACT TOX-001, LRN-U2103 This study was conducted in compliance with United States Food and Drug Administration (FDA) Good Laboratory Practice (GLP) Regulations 21 CFR Part 58, with the exceptions in the bulleted list below. Exceptions to GLP compliance: 0 There were two study directors in this study. This study was designed as two separate studies. The in-life phase study was considered to end at the generation and shipment of specimens. The analytical study was considered to start at the receipt of these specimens for analysis. This resulted in having two separate study directors, one for each phase of the same study. However, since the technical performance of each phase was entirely separate, no effect is expected from this exception. 0 Sample storage stability will not be determined. 0 Characterizationof the analytical standards is underway, but has not yet been completed (21 CFR 58.105 (a)). Lot No. 59905 of the surrogate, THPFOS, will not be characterized for purity since quantities of this standard are exhausted. 0 The electronic data systems in use have not been validated and there is not an electronic audit trail of corrections currently available (21 CFR 58.130 (e)). Authenticated hard copies of chromatograms and associated documents will be considered as the original raw data. 0 Some changes in the raw data entries were not all made in accordance with 21 CFR 58.130 (e). 0 Some reagents and solutions did not have expiration dates on the labels, as required according to 21 CFR 58.83. (See next page for signatures) 3M Environmental Laboratory 3M Environmental Laboratory Page 3 Page 3 3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 GLP Compliance Statement (continued) Analytical Report: FACT-TOX-001 LRN-U2103 Analytical Report: FACT TOX-001 LRN-U2103 7L2. - John L. Butenhoff, Ph.D., Stud$ Director 6 .-' 7 C L Marvin T. Case, D.M.V., Ph.D., Sponsor Representative &/ m/ Date !bq Date 05\31 /O/ Kristen'J. Hansen, Ph .D., Principal Analytical lnvestigator Date William K. Reagen, Ph.D., Laboratory Manager Date 3M Environmental Laboratory 3M Environmental Laboratory Page 4 Page 4 3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Analytical Report: FACT TOX-001 LRN-U2103 GLP Study-Quality Assurance Statement Analytical Laboratory Report Title: Determination of the Presence and Concentration of PFOS, PFOSA, PFOSAA, EtFOSE-OH, M556, and PFOSEA in Serum and Liver Samples of Crl:CD@(SD)IGS BR Rats Exposed to N-Ethyl Perfluorooctanesulfonamido Ethanol Study Identification Numbers: T-6316.1, FACT TOX-001, LRN-U2103 This study has been inspected by the 3M Environmental Laboratory Quality Assurance Unit (QAU) as indicated in the following table. The findings were reported to the study director and laboratory management. Inspection Dates I Phase Date Reported to Management I Study Director 4 / 7 / 0 0 4 1 1/00 In-phase 5/8/00 5/8/00 1/15/01-2/4/0 1 Data 2/5/0 1 2/5/0 1 I I 1 3/23/01, 3/26/01-3/30/01 3/26/01-3/27/0 1, 3/29/01- I I I 3/30/01,4/2/01-4/5/01 Data Data 3/30/01 4/6/01 3/30/01 4/6/01 4/3/01-4/6/0 1, 4112/01-4113101, 4/17/0 1 Data 4/18/0 1 4/18/0 1 5/3/01,5/7/01-5/10/01 , 5/15/01-5/16/0 1 Draft Report 5/16/01 5/16/01 QAU Representative ~ Date 3M Environmental Laboratory 3M Environmental Laboratory Page 5 Page 5 3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 Table of Contents Analytical Report: FACT-TOX-001 LRN-U2103 Analytical Report: FACT TOX-001 LRN-U2103 GLP Compliance Statement ............................................................................................. 3 GLP Study-Quality Assurance Statement ...................................................................... 5 Study Personnel and Contributors.................................................................................... 8 Introduction and Purpose.................................................................................................. 9 Test System ................................................................................................................. 9 Specimen Collection and Analysis ............................................................................... 10 Specimen Receipt and Maintenance................................................................................ 10 Chemical Characterization of the Reference Materials..................................................... 12 Dose Confirmation Analyses ........................................................................................ 12 Method Summaries........................................................................................................... 13 3M Environmental Laboratory ...................................................................................... 13 Preparatory Methods............................................................................................... 13 Analytical Methods .................................................................................................. 14 Analytical Equipment............................................................................................... 14 Data Quality Objectives and Data Integrity ....................................................................... 15 Data Summary. Analyses. and Results............................................................................. 15 Summary of Quality Control Analyses Results for PFOS. PFOSA. PFOSAA. M556. EtFOSE-OH and PFOSEA........................................................................................... 16 Statement of Data Quality............................................................................................ 18 Summary of Sample Results........................................................................................ 19 Statistical Methods and Calculations ................................................................................ 19 Statement of Conclusion................................................................................................... 19 References ....................................................................................................................... 19 Appendix A: Chemical Characterization and Control Matrices.......................................... 20 Appendix B: Protocol, Amendments and Deviations......................................................... 21 Appendix C: Extraction and Analytical Methods ............................................................... 12025 Appendix D: Data Summary Tables.................................................................................. 12835 Appendix E: Data Spreadsheets....................................................................................... 13968 Appendix F: Example Calculations ................................................................................... 23784 Appendix G: Interim Certificates of Analysis..................................................................... 32885 Appendix H: Report Signature Page................................................................................. 33190 3M Environmental Laboratory 3M Environmental Laboratory Page 6 Page 6 3M Medical Department Study: T6316.1 3M Medical Department Study: 1-6316.1 List of Tables Analytical Report: FACT-TOX-001 LRN-U2103 Analytical Report: FACT TOX-001 LRN-U2103 Table 1.Test System Population Demographicsfor Study 6329-212 .............................. 9 Table 2. Specimen Collection for FACT TOX-001 ............................................................ 10 Table 3. Characterization of the Analytical Reference Standards in Study FACT TOX-001 .............................................................................................................. 12 Table 4.Target Ions Monitored in 3M Laboratory Analyses.............................................. 15 Table 5. Matrix Spikes-Sera .......................................................................................... 17 Table 6. Matrix Spikes-Liver ........................................................................................... 18 Table 7. Characterizationof the Control Matrices Used for Sera Analyses in Study FACT TOX-001.................................................................................................... 20 Table 8. Characterizationof the Control Matrices Used for Liver Analyses in Study FACT TOX-001.................................................................................................... 20 Table 9. Characterization of Test Article in Study FACT TOX-001 ................................... 20 Table 10. FACT TOX-001 Data Summary of PFOS Concentration-Serum (pg/mL) .......21486 Table 11. FACT TOX-001 Data Summary of PFOSA Concentration-Serum (pg/mL).....21587 Table 12. FACT TOX-001 Data Summary of PFOSAA Concentration-Serum (pg/mL) ..21688 Table 13. FACT TOX-001 Data Summary of EtFOSE-OHConcentration-Serum (pg/mL) ................................................................................................................ 12879 Table 14. FACT TOX-001 Data Summary of M556 Concentration-Serum (pg/mL)........12980 Table 15. FACT TOX-001 Data Summary of PFOSEA Concentration-Serum (pg/mL) ..2191 Table 16. FACT TOX-001 Data Summary of PFOS Concentration-Liver (pg/g)............. 13902 Table 17. FACT TOX-001 Data Summary of PFOSA Concentration-Liver (pg/g) ..........13193 Table 18. FACT TOX-001 Data Summary of PFOSAA Concentration-Liver (pg/g) ........13294 Table 19. FACT TOX-001 Data Summary of EtFOSE-OH Concentration-Liver (pg/g)...31395 Table 20. FACT TOX-001 Data Summary of M556 Concentration-Liver (pg/g) .............31496 Table 21. FACT TOX-001 Data Summary of PFOSEA Concentration-Liver (pg/g) ........13957 3M Environmental Laboratory 3M Environmental Laboratory Page 7 Page 7 3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 Study Personnel and Contributors Analytical Report: FACT-TOX-001 LRN-U2103 Analytical Report: FACT TOX-001 LRN-U2103 Study Director John L. Butenhoff, Ph.D. 3M Corporate Toxicology - Medical Department 3M Center Building 220-2E-02 St. Paul, MN 55144-1000 651-733-1962 Sponsor Marvin T. Case, D.V.M., Ph.D. 3M Corporate Toxicology - Medical Department 3M Center Building 220-2E-02 St. Paul, MN 55144-1000 Analytical Chemistry Laboratories Liver and Serum Analyses 3M Environmental Laboratory Kristen J. Hansen, Ph.D., Principal Analytical lnvestiga tor 3M Lab Contributing Personnel David R. Barnidge, Lisa A. Clemen Lisa A. Dick, Ph.D. Rhonda S. Dick* Kelly J. Dorweiler* Mark E. Ellefson Sara E. Estes* Tina M. Galloway Barb A. Gramenz* Sarah A. Heimdal* Cari S. Hewitt* Ph.D.* "Contract lab professional service employees Marlene M. Heying* Joy D. Jenkins Harold 0. Johnson Ognjenka Kruplijanin* Sally A. Linda* Ian A. Smith* Kathleen M. Stock* Kelly J. (Kuehlwein)*Swartout Anh-Dao Vo Bob W. Wynne* Richard D. Youngblom* Location of Archives All original raw data, protocol, and analytical report have been archived at the 3M Environmental Laboratory. The test article and analytical reference standard reserve samples, as well as the specimens pertaining to the analytical phase of this study are archived at the 3M Environmental Laboratory. 3M Environmental Laboratory 3M Environmental Laboratory Page 8 Page 8 3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 Introduction and Purpose Analytical Report: FACT-TOX-001 LRN-U2103 Analytical Report: FACT TOX-001 LRN-U2103 The purpose of the study is to determine the presence and concentration of PFOS, PFOSA, PFOSAA, EtFOSE-OH, M556, and PFOSEA in serum and liver specimens collected from Covance Study No.: 6329-212 titled: 104-Week Dietary Carcinogenicity Study with Narrow Range (98.1%) N-Ethyl Perfluorooctanesulfonamido Ethanol in Rats. The Covance in-life study was initiated on January 21, 1998. The analytical phase of the study was initiated on April 9, 1998. Test System A total of 410 males and 410 female rats were used as the test system. Table 1 outlines the rat population demographics and dosage levels for study 6329-212. The test system species and strain selected was the Crl:CD@(SD)IGS BR rat received from Charles River Laboratories, Inc., identified using an implanted microchip device. At the initiation of treatment the rats were approximately 45-51 days old and weighed between approximately 100-31 0 g. Table 1. Test System Population Demographicsfor Study 6329-212 a Groups 5 and 7 were terminated during week 8 of treatment due to toxjcily. Group 6 was plxedon recoveryafter 52 weeks of treatment. 3M Environmental Laboratory 3M Environmental Laboratory Page 9 Page 9 3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Analytical Report: FACT TOX-001 LRN-U2103 Specimen Collection and Analysis Specimens were collected by Covance (study 6329-212) and sent to the 3M Environmental Laboratory for analysis. Table 2 lists the sampling intervals and types of specimens that were collected for this study. Table 2. Specimen Collection for FACTTOX-001 The total number and type of specimens collected for analyses in the analytical phase of this study are presented below. Specimens Collected from Study Groups 1through 5 (through 2/01/00): Serum Specimens-524 specimens (423 samples were analyzed.) Liver Specimens-265 specimens (All 265 samples analyzed.) Specimens Collected from Study Group 6 (Recovery) on 2/01/00: Serum Specimens-22 specimens (All 22 samples analyzed.) Liver Specimens-22 specimens (All 22 samples analyzed.) Liver and sera specimens were shipped to the 3M Environmental Laboratory frozen and on dry ice. Sera and liver samples were extracted beginning on April 23, 1998 using an ion pairing reagent and either ethyl acetate or methyl-tert-butyl ether (MtBE). Liver samples were homogenized prior to the extraction procedure. Sample extracts were analyzed using high-pressure liquid chromatography-electrosprayAandem mass spectrometry (HPLC-ESMSMS) in the multiple reaction monitoring mode. PFOS, PFOSA, PFOSAA, EtFOSE-OH, M556, and PFOSEA levels were evaluated by external calibration using extracted curves. Analytical details are included in this report. Specimen Receipt and Maintenance The 3M Environmental Laboratory received liver and serum specimens collected at predeterminedtime points during and at the end of the in-life phase of Covance Study 6329-212 from February 1998 through February 2000 from Covance. All specimens were received frozen on dry ice and were immediately transferred to storage at -20C *IO"C. 3M Environmental Laboratory 3M Environmental Laboratory Page 10 Page 10 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 3M Medical Department Study: T-6316.1 Analytical Report: FACT TOX-001 LRN-U2103 Control matrices used in liver and sera analyses performed during TOX-001 were obtained from commercial sources and are presented in Appendix A (see Tables 7 and 8).Samples analyzed at the 3M Environmental Laboratory will be maintained for a period of 10 years and will be stored at the laboratory at -20Ck10"C to -80Ci2O"C. 3M Environmental Laboratory 3M Environmental Laboratory Page 11 Page 11 3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Analytical Report: FACT TOX-001 LRN-U2103 Chemical Characterization of the Reference Materials Chemical characterization information on the analytical reference materials used in this study is presented in tabular form below. Table 3. Characterization of the Analytical Reference Standards in Study FACT TOX-001 Reference Standard/ Formula Potassium Perfluorooctanesulfonate CeFi7S03-K+ N-Ethyl Perfluorooctanesulfonamidoethyl alcohol C~FI~SOZN(GH~)CHZCHZOH Sodium Perfluorooctanesulfonylamido(ethyl) acetate CEFI~SOZN(CH~CH~KHZCOO-N~ Perfluorooctanesulfonylamido(ethyl)acid CEFI~SO~N(CHZCH~)CHZCHOO- Perfluorooctanesulfonylamide CeFi7SOzNHz Perfluorooctanesulfonylethylamide CsFi7SOzNHCHzCH3 M556 C8Fi7S02N(H)CH2COOH lH, lH, 2H, 2HTetrahydroperfluorooctanesulfonic acid CEH~FI~SO~H 'This lot is exhausted and cannot be characterized. N R - N o t recorded N A - N o t applicable TBD-To be determined Acronym Source Expiration Storage Chemical Physical Date Conditions Lot Number Description Purity 3M Oml/O1 Ambient temperature KPFOS" 3M 2010 Ambient temperature 171 'ght 86.4% powder 193 W hte 88.0% crystals Ambient 3M 2010 temperature 215 Whitepowder TBD' 3M EtFOSE-OH 3M I I I 1 1 1 3M PFOSMb I I I t$:!zre I I I 3M 201 0 11/26/01 Ambient temperature Ambient temperature 01/01/2010 $t$er 936 Unknown (SD013) 617 Amberwaxy solid Amberwaxy solid NA* 88.9% TBD' 01/01/2010 NB112999-99 Tanwaxysdid TBD' Ambient 3M 01/01/2010 temperature E353 Amber to brown waxy sdid TBDC PFOSA 3M 201 0 Ambient NR NR temperature (TN-A-1886) TBD' I I I I PFOSEA 3M 01/01/2010 temAmpebriaetnutre 3M 01/01/2010 temAmpebireantut re L15709 529 Light YdlW TBDc I ""zidWI waxysdid TBD' M556 THPFOS 3M 01/01/2010 temAmpebireantut re NB113047-80 White powder TBD ICN 01/01/201 O Ambient temperature Ambient ICN 01/01/2010 temperature 59909 53406 Brownpowder Brownwaxy Solid NA* TBDc "rarget analyie is C8Fl7SO2{(CH2CH3)(CH2COO]) 'Unless othemise indicated, at the time of quantitation, the purity for all analyies was assumedto be 1W/. Dose Confirmation Analyses Dose preparation methods and analysis were performed by Covance, using a validated analytical method provided by the Sponsor (MP-M312-MA), and are reported separately (Reference Covance 6329-212). 3M Environmental Laboratory 3M Environmental Laboratory Page 12 Page 12 3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 Method Summaries Analytical Report: FACT-TOX-001 LRN-U2103 Analytical Report: FACT TOX-001 LRN-U2103 Following is a brief description of the methods used during this analytical study by the 3M Environmental Laboratory. Detailed descriptions of the methods used in this study are located in Appendix C. Data collected prior to November 1999 was reworked in 2000 to accommodate improvements in data reduction methods. Both the original and "reworked" data are archived; reworked data is presented in the final results. The improved methods are documented in the form of method modifications. As the present study progressed, more advanced methods evolved and these methods were used with deviations until amendments to the protocol were written. Protocol and method deviations are located in Appendix B of this report. 3M Environmental Laboratory PREPARATORMYELHODS FACT-M-1.O. "Extraction of Potassium Perfluorooctanesulfonateor Other Anionic FluorochemicalSurfactants from Liver for Analysis Using HPLC-Electrospray/Mass Spectrometry." This method was used for week 4, week 8, and week 14 samples. FACT-M-3.0, "Extraction of Potassium Perfluorooctanesulfonateor Other Anionic FluorochemicalSurfactants from Serum for Analysis using HPLC-Electrospray/Mass Spectrometry." This method was used for week 4, week 8, and week 14 samples. ETS-8-4.1, "Extraction of Potassium Perfluorooctanesulfonate or other Fluorochemical Compounds from Serum for Analysis using HPLC-Electrospray/MassSpectrometry." This method was used for week 27, week 53, and week 105 samples. ETS-8-6.0, "Extraction of Potassium Perfluorooctanesulfonateor other Fluorochemical Compounds from Liver for Analysis using HPLC-Electrospray/MassSpectrometry." This method was used for week 53 and week 105 samples. An ion-pairing reagent was added to the sample and the analyte ion pair was partitioned into ethyl acetate (FACT-M-1.O and FACT-M-3.0) or MtBE (ETS-8-4.1 and ETS-8-6.0). The extract was transferred to a centrifuge tube and put onto a nitrogen evaporator until dry. Each extract was reconstituted in 1.OmL of methanol, and then filtered through a 3 cc plastic syringe attached to a 0.2 pm nylon filter into glass autovials. 3M Environmental Laboratory 3M Environmental Laboratory Page 13 Page 13 3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Analytical Report: FACT TOX-001 LRN-U2103 ANALYTICAL METHODS FACT-M-2.0, "Analysis of Liver Extracts for Fluorochemicals Using HPLC-Electrospray/Mass Spectrometry". FACT-M-4.0, "Analysis of Fluorochemicals in Serum Extracts Using HPLC-Electrospray/Mass Spectrometry". ETS-8-5.1, "Analysis of Potassium Perfluorooctanesulfonateor other Fluorochemicals in Serum Extracts Using HPLC-Electrospray/MassSpectrometry". ETS-8-7.0, "Analysis of Potassium Perfluorooctanesulfonateor other Fluorochemicals in Liver Extracts Using HPLC-Electrospray/MassSpectrometry". The analyses were performed by monitoring one or more product ions selected from a single primary ion characteristic of a particular fluorochemical using HPLC-ES/MS/MS. For example, molecular ion 499, selected as the primary ion for PFOS (C8FI7SO3-)analysis, was fragmented further to produce ion 99 (FS03-). The characteristic product ion 99 was monitored for quantitative analysis. ANAL YTICAL EQUIPMENT The actual analytical equipment settings used in the present analytical phase of this study varied slightly during actual data collection. The following is representative of the settings used during the analytical phase of this study. Liquid Chromatograph: Hewlett-Packard@Series 1100 Liquid Chromatograph system Analytical column: Keystone@BetasilTMCIS2x50 mm (5 pm) Column temperature: Ambient Mobile phase components: Component A: 2mM ammonium acetate Component B: methanol Flow rate: 300 pUmin Injection volume: 10 p L Solvent Gradient: 13.5 minutes Time (minutes) %B 0.0 40% 8.5 90% 11.0 90% 12.0 40% 13.5 0% 3M Environmental Laboratory 3M Environmental Laboratory Page 14 Page 14 3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Analytical Report: FACT TOX-001 LRN-U2103 Mass Spectrometer: Micromass@APVMass Spectrometer Quattro llrM Triple Quadrupole system Software: Mass LynxrM2.3,3.1,3.2,3.3,3.4 Cone Voltage: 30-60 V Collision Gas Energy: 25-45 eV Mode: Electrospray Negative Source Block Temperature: 150C ~ 1 0 C Electrode: Z-spray Analysis Type: Multiple Reaction Monitoring (MRM) Table 4. Target Ions Monitored in 3M LaboratoryAnalyses I Target Analge I~~ Primary Ion (AMU) I P r o d u x o ( A M U ) l I PFOS I 499 I 80,99,130 I PFOSA 498 78 I PFOSAA 1 584 I 83,169 I EtFOSE-OH 630 59 PFOSEA 526 65 M556 556 65,78,831,69 THPFOS 427 80 Data Quality Objectives and Data Integrity The following data quality objectives were indicated in the method performance section of ETS- 8-5.1, Analysis of Potassium Perfluorooctanesulfonateor Other Fluorochemicals in Serum Extracts Using HPLC-Electrospray/MassSpectrometry and ETS-8-7.0, Analysis of Potassium Perfluorooctanesulfonateor Other Fluorochemicals in Liver Extracts Using HPLC-Electrospray/Mass Spectrometry: 0 Linearity: The coefficient of determination (r2) equal to or greater than 0.980. 0 Limits of Quantitation (LOQ): The LOQ is equal to the lowest acceptable standard in the calibration cuwe. 0 Acceptable Precision: Precision is better than 30% for the method. 0 Acceptable Spike Recoveries: 70-1 30% Data Summary, Analyses, and Results Data quality objectives for the analytical phase of this study outlined in the 3M Environmental Laboratory protocol for FACT TOX-001 (see Appendix B) were met with the exceptions noted in this report. See Appendix B for deviations. 3M Environmental Laboratory 3M Environmental Laboratory Page 15 Page 15 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 3M Medical Department Study: T-6316.1 Analytical Report: FACT TOX-001 LRN-U2103 Summary of Quality Control Analyses Results for PFOS, PFOSA, PFOSAA, M556, EtFOSE-OHand PFOSEA Linearity: The coefficient of determination (r2)of the standard curve was 20.980. Calibration Standards: Quantitation of the target analytes was based on linear regression analysis weighted l / x of a single, opening or two bracketing extracted matrix curves for each group of samples. Rat and rabbit sera were used for matrix curves for sera analyses, while rabbit liver was used for matrix curves for liver analyses. High or low points on the curve may have been deactivated to provide a better linear fit over the curve range most appropriate to the data. Low curve points with peak areas less than two times that of the extraction blanks were deactivated to disqualify a data range that may have been significantly affected by background levels of the analyte. Occasionally, a single mid-range curve point that was an obvious outlier may have been deactivated. Quantitation of each analyte was based on the response of one or more specific product ion@)using the multiple reaction monitoring mode of the instrument (see Appendix C, Analytical Methods). Calibration standards were prepared to run, undiluted, approximately within the linear range of the instrument (approximately 51000 ng/g). Limits of Quantitation (LOQ): The LOQ is equal to the lowest acceptable standard in the calibration curve (defined as a standard within *30% of the theoretical value), and is at least two times the analyte peak area detected in the surrogate matrix blanks. (see Appendix D). Blanks: All blanks were below the limit of quantitation for the compounds of interest, except PFOSA Week 8 H 2 0+ liver Blk-2 (0.0663 pg/g and 0.0414 pg/g) and PFOS Week 105 liver- Blk-2 (0.0163). To simplify analyses that were complicated by endogenous levels of fluorochemicals in unexposed rat sera and liver, rabbit sera and liver were selected as suitable surrogate matrices. Precision, Instrumental: Instrumental precision was determined by replicate injections of a single serum extract. Instrumental precision was determined for PFOS, PFOSAA, and M556; variation was less than 7.0 Yofor all targeted analytes. Matrix Spikes: Sera-Matrix spike samples were prepared from control rat sera along with each batch of sera samples. Samples were spiked between approximately 75-250 ng/mL, levels that approximate the levels detected in the Group l-Group 5 samples, depending upon the analyte. All spikes were prepared to run, undiluted, within a ten-fold limit of the linear range of the calibration curve. For some analytes, samples for some high dose animals may be much higher than the range of these prepared spikes. Sera matrix spike recoveries are presented in tabular form in Table 5. 3M Environmental Laboratory 3M Environmental Laboratory Page 76 Page 16 3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Analytical Report: FACT TOX-001 LRN-U2103 1 I 1 Table 5. Matrix Spikes-Sera # of Spikes * Averagespike Recovery SD PFOS 1 PFOSA 1 28 I 9 5 i 2 5 % 1 28 103 2 20% PFOSAA 28 111 i 2 4 % EtFOSE-OH 22 39 i 29% M556* PFOSEA 22 126 i 22% 22 63 * 14% Range 65183% 72-1 43% 57-1 86% 18-1 27% 86-1 53% 3747% # of Spikes Deviating > &o% and < *50% 1 3 I 4 3 0 8 8 II 1 1 0 1 20 3 6 3M Environmental Laboratory 3M Environmental Laboratory Page 17 Page 17 3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Analytical Report: FACT TOX-001 LRN-U2103 PFOS # of Spikes 24 Average Spike Recovery f SD 90 * 61% Range 0-1 96% # of Spikes Deviating > *30% and < *50% 4 # of Spikes Deviating > &50% 9 PFOS, revised (') 18 119 * 38% 6&196% 4 3 PFOSA 18 94 f 27% 26-128% 0 2 1 I I I I I PFOSA, revised (') 16 102 * 17% 71-128Yo 0 0 PFOSAA 20 86 f 44% 0-1 47% 1 4 1 1 PFOSAA, revised (') 105*19% 174147% 1 0 MFOSE-OH 16 133 f 166% 25567% 1 8 EtFOSE, revised ('I 14 148 173% 26-567% 1 6 M556* 12 102 * 15% 85141% 1 0 PFOSEA 10 97 * 12% 80-117% 0 0 Surrogates: The surrogate (THPFOS) was added to all samples and standards. THPFOS was not used for quantitation, but was used to monitor for gross instrument failure. After 11/04/99, the surrogate response of each analytical run was verified to determine that it did not vary more than &50% from the mean within each analytical run. Deviations of greater than f 50% are noted in the results table. Statement of Data Quality It is not possible to verify true recovery of endogenous analyte from tissues without radio-labeled reference material. The only measurement of accuracy available at this time, matrix spike studies, indicate that the sera and liver data can be considered accurate to within one standard deviation of the average fortified samples recovery. For example, in liver, PFOS data is accurate to 119 f 38%. Please see bolded values in Tables 4 and 5 for specific values relating to data quality. The results of quality control analyses (curve fit, CCVs, and MS/MSDs) for EtFOSE-OH and PFOSEA were inconsistent and indicate that data presentedfor these two analytes should be 3M Environmental Laboratory 3M Environmental Laboratory Page 18 Page 18 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 3M Medical Department Study: T-6316.1 Analytical Report: FACT TOX-001 LRN-U2103 considered to be qualitative only. Values for these two analytes are presented in data tables in the spirit of full disclosure, but should not be used in any quantitative assessment of the data. Summary of Sample Results Some PFOS results (those obtained using lot 171) have been corrected for purity of the analytical reference material. Uncorrected results are noted in the data tables. 0 Samples from Control Animals: The target analytes were often detected in the sera and liver samples from the control animals. These levels were usually lower than those found in the low dose test animals. 0 Samples from Dosed Animals: In general, levels of the target analytes present in the sera and liver samples from the test animals increased with dose group, with the exception of Group 6, which was taken off the compound after 53 weeks. Detailed sample data tables are presented in Appendices D and E. Statistical Methods and Calculations Statistical methods were limited to the calculation of means and standard deviations. See Appendix F for example calculations used to generate the liver and serum sample data in FACT TOX-001, Statement of Conclusion Under the conditions of the present study, PFOS, PFOSA, PFOSAA, EtFOSE-OH, M556, and PFOSEA were observed in the sera and liver of rats dosed with N-Ethyl Perfluoroctanesulfonamido Ethanol during the in-life phase of the study. References Covance Study No.: 6329-212, 104-Week Dietary Carcinogenicity Study with Narrow Range (98.1%) N-Ethyl PerfluoroctanesulfonamidoEthanol in Rats 3M Environmental Laboratory 3M Environmental Laboratory Page 19 Page 19 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 3M Medical Department Study: T-6316.1 Analytical Report: FACT TOX-001 LRN-U2103 Appendix A: Chemical Characterization and Control Matrices Table 7. Characterization of the Control Matrices Used for Sera Analyses in Study FACT TOX-001 Control Matrix I RatSerum 1 Rat Serum 1 Rabbitserum 1 ~ Source Sigma Sigma Sigma Expiration Date 201 0 2010 2010 Storage Conditions -20C k10"C -20C *I 0C -20C *I 0C Chemical Lot # 17H9306 19H89291 47H4641 Physical Description I NR-nd recorded Rat Serum I I Rat Serum Rabbit Serum I I Control Matrix Rabbit Liver Rabbit Liver Rabbit Liver Rabbit Liver Rabbit Liver Source Covance Laboratories, Inc.* Covance Laboratories, Inc.' Covance Laboratories, Inc.' Covance Laboratories, Inc.' Unknown Expiration Date 2010 201 0 12/99 2010 12/01/99 Storage Conditions -20C *1O"C -20C *I 0C -20C *I 0C -20C *I 0C -20C *lO"C Chemical Lot # FOOOI 4 F00008 F00007 FOOOI 6 NR (TCR-99062-22) Physical Description Rabbit Liver Rabbit Liver I Rabbit Liver 1 Rabbit Liver Rabbit Liver I NR-not recorded T h e controlsource is listedas CHW (Coming HazeltonWisconsin) in the rawdata This is the former name of C m c e Laboratories. Inc. Table 9. Characterization of Test Article in Study FACT TOX-001 Chemical Name Source Expiration Date Storage Conditions I Chemical Lot # Physical Description Purity Et FOSE-OH N-Ethyl Perfluorooctanesulfonamido ethanol 3M 1 1/26/01 Ambient temperature I 30035,30037,30039 1 Waxy solid 97.4%' 3M Environmental Laboratory 3M Environmental Laboratory Page 20 Page 20 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 3M Medical Department Study: T-6316.1 Analytical Report: FACT TOX-001 LRN-U2103 ~ Appendix B: Protocol, Amendments and Deviations 3M Environmental Laboratory 3M Environmental Laboratory Page 21 Page 21 3M Medical Department Study: T6316.1 c o v m THE DWELOPYLNTSERVICESC W M Analytical Report: FACT-TOX-001 LRN-U2103 3M St. Paul, Minnesota PROTOCOL Study 'ICtle: 104-Wezk Dietary CarcinogenicityStudy with Narrow Range (98.1%) N-Ethyl PerfluorooctanesulfonamidoEthanol in Rats Date: January 21,1998 Performing Laboratory: Covance Laboratories Inc. 3301 Kiusman Boulevard Madison, Wisconsin 53704 Laboratory Study Identification: Proposal No. 6777 C O V ~63~29C-2~12 3M Environmental Laboratory Page 22 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 C O V ~ ~6N32X9-212 Page 2 Study 104-Week Dietary Carcinogenicity Study with Narrow Range (98.1%) N-Ethyl Perfluorooctanesulfonamido Ethanol in Rats Purpose To assess the carcinogenicity of the test material when administered in the diet to rats for at least 104 weeks Sponsor 3M Toxicology Services Building 220-2E-O2,3M Center St. Paul, Minnesota 55144-1000 Study Monitor Andrew M. Seacat, PhD 3M Telephone No.: 612.575.3161 Facsimile No.: 612.733.1773 Study Location Covance Laboratories Inc. 3301 Kinsman Boulevard Madison, Wisconsin 53704 Mailing Address: PO Box 7545 Madison, Wisconsin 53707 Study Director Peter J. Thomford, PhD Covance Laboratories Inc. Telephone No.: 608.241.7207 Facsimile No.: 608.242.2736 3M Environmental Laboratory Page 23 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Toxicologist Thomas E. Ryan,BS Covance Laboratories Inc. Covance 6329-212 Page 3 Proposed Study Timetable In-Life Start Date: January 26, 1998 In-Life End Date: January 31,2000 Regulatory Compliance This study will be conducted in compliance with the Food and Drug Administration Good Laboratory Practice Regulations as set forth in Title 21 of the US Code of Federal Regulations, Part 58, issued December 22, 1978 (effective June 20, 1979), and with any applicable amendments. Animal Care and Use Statement All procedures in this protocol are in compliance with the Animal Welfare Act Regulations, 9 CFR 1-4. In the opinion of the Sponsor and study director, the study does not unnecessarily duplicate any previous work. Quality Assurance The protocol, study conduct, and final report will be audited by the Covance Qu&ty Assurance Unit (QAU). The proliferation cell nuclear antigen evaluation, data, and report will be audited by the QAU of Pathology Associates InternationaL Test Material Identification T-6316 (Narrow Range N-Ethyl PeffluorooctanesulfonamidoEthanol, NEtFOSE) Lot Numbers The lot numbers will be maintained in the raw data. Purity 98.1% NEtFOSE (w/w) 3M Environmental Laboratory Page 24 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Covance 6329-212 Stability Responsibilityof the Sponsor Storage Conditions At room temperature Characteristics Information on synthesis methods, composition, or other characteristicsthat define the test material is on file with the Sponsor. Reserve (Archive) Samples A reserve sample (approximately5 g) will be taken and stored at room temperature. This sample will be transferred to the Sponsor after completion of the in-lifephase to be retained in accordance with 21 CFR 58.195. Disposition of Test Material After authorization from the Sponsor, any remaining test material will be returned to: Andrew M. Seacat, PhD 3M Toxicology Services Building 220-2B-O2,3M Center St. Paul, Minnesota 55144-1000 Telephone No.: 612.575.3 161 Facsimile No.: 612.733.1733 Animals species Rat Strain . Crl:CD@(SD)BR 3M Environmental Laboratory Page 25 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Source Charles River Laboratories, Inc., Raleigh, North Carolina Covance 6329-2 12 Page 5 Age at Initiation of Treatment Preferably 6 weeks of age, but not more than 8 weeks of age Weight at Initiation of Treatment 100to 300 g Number and Sex 410 males and 4 10females Identification Implantable microchip identification device Husbandry Housing Individual (may be group-housed during acclimation) Diet Certified Rodent Diet #5002 (PMJ? Feeds, Inc.) ad libitum, unless otherwise specified. The diet is routinely analyzed by the manufacturer for nutritional components and environmental contaminants. Specified nutrient and contaminant analyses are on file at Covance-Madison. Water Ad libitum. Samples of the water are routinely analyzed for specified microorganisms and environmental contaminants. The results are on file at Covance-Madison. Contaminants There are no known contaminants in the diet or water at levels that might interfere with this study. 3M Environmental Laboratory Page 26 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Covance 6329-2 12 Page 6 Environment Environmental controls for the animal room will be set to maintain 18 to 26"C, a relative humidity of 30 to 70%, a 12-hour IightllZhour dark cycle, and minimum of 10 room air changeshour. The lighvdark cycle may be interrupted to accommodate study-related activities. Acclimation At least 1 week Randomization Selection of animals for the study will be based on body weights, c h c a l observations, and other data as appropriate. Animals wjll be assigned to treatment groups using a computerized blocking procedure designed to achieve body weight balance with respect to treatment groups. At the time of randomization,the weight variation of the animalsof each sex used will not exceed & standard deviations of the mean weight, and the mean body weight for each group of each sex will not be statisticallydifferent at the 5.0% probability level. Justification Rats historically have been used in safety evaluation studies and are recommended by appropriate regulatory agencies. 3M Environmental Laboratory Page 27 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Group Designations and Dietary Levels Covance 6329-2 12 Page 7 Number of Animals Dietary Levels Group Male Female (ppm NEtFOSE)" 1 (ControI)b-c*d 70 70 0 2 (Low)" 60 60 3 3 (Mid)c 60 60 30 4 (Mid-HighYd 70 70 100 5 (HighYd 70 70 300 6 (Mid-High Recovery)" 40 40 100 7 (High Recovery)" 40 40 300 a T-6316 is 98.1% n-ethyl perfluorooctanesulfonamido ethanol (NEtFOSE); dose levels are expressed as ppm of NEtFOSE. b The control animalswill receive the basal diet only. c Five animals/sex m Groups 1 through 5 will be sacrificed during Weeks 3 and 14 for hepatocellular proliferationrate measurements and biochemical analyses (palmitoyl-CoA oxidation). d Ten animals/sex in Groups 1,4, and 5 will be designated as interim sacrifice animals and will be sacrificed after at least 78 weeks of treatment. e Animals in Groups 6 and 7 will be treated for at least 78 weeks,then treatment will be discontinued, and the animalswill be observed for reversibility, persistence, or delayed occurrence of toxic effects for at least 26 weeks posttreatment. During recovery, the animalswill receive basaldiet only. Dosing Procedures Method ofAdministration Dietary. Animals in Groups 1 through 5 willreceive test diet for at least 104 weeks. Animals in Groups 6 and 7 will receive test diet for 78 weeks only. Reason for Dosing Route The potential human exposure is by the oral route. Dose Preparation Before initiation of treatment, alldose preparations will be mixed once, and low- and high-dose preparations will be mixed four times for determination of homogeneity and stability. 3M Environmental Laboratory Page 28 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Covance 6329-212 During treatment, all dose preparations will be mixed at least once every 4 weeks according to the study-specificmixing procedure developed by Covance. Dose concentrations will be based on the NEtFOSE content as supplied; the Sponsor has stated the T-6316 is 98.1% NEtFOSE (w/w) as supplied. All dose preparations will be stored at room temperature. Retention Samples Samples (approximately 100g) will be taken from each dose preparation during the in-life phase and stored at room temperature. Unless used for analyses, these samples will be discarded at least 1 month after completion of the in-life phase. Dose Analyses By Covance using a method supplied by the Sponsorand validated by Covance Homogeneity Homogeneity will be determinedfor all dose level preparations once pretest and for the preparations for Weeks 1 through 3; for two additional pretest high-dose level preparations; and for four additional pretest low-dose level preparations. One sample (approximately 100 g) each from the top, middle, and bottom of the dose preparations mixed for homogeneity analyses will be collected, divided into three subsamplesfor extraction and analysis, and analyzed for test material content. All samples will be stored at room temperature until analyzed within 7 days of mixing. Homogeneity analysis will be repeated if batch size changes by more than 30%. Stability Four sets of samples (approximately 100g each) will be taken from the low- and high-dose level concentrations of diet preparations mixed pretest to establish stability. One set will be analyzed on the day of mixing. One set will be stored at room temperature for at least 19 days, then analyzed. The third set will be stored at room temperature after at least 32 days, then analyzed. The remaining set will be stored in a freezer set to maintain -10 to -30C for 8 weeks, then analyzed. 3M Environmental Laboratory Page 29 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Covance 6329-212 Page 9 In addition, samples (approximately 100 g each) wjll be taken from the low- and high-dose level concentrations of diet preparations mixed pretest. The samples will be stored at room temperature for 5 days, then for 7 days under animal room conditions (room temperature in a feeding container), then analyzed. Dose Cofirmation Samples (approximately 100 g each) will be collected from all dose preparations and analyzed in duplicate. Homogeneity samples collected from the middle of the dose preparations for Weeks 1 through 3 will be used for dose confirmation results. All samples will be stored at room temperature until analyzed. Observation of Animals Clinical Observations Each animal will be observed twice daily (a.m. and p . a ) for mortality and moribundity, recording findings as they are observed. Once prior to treatment and weekly thereafter, each animal will be removed from its cage and examined; abnormal findings or an indication of normal will be recorded. The following information on each grossly visible or palpable mass will be recorded. time of onset location size (smallor large) appearance progression Body Weights Prior to treatment (at randomization), weekly for Weeks 1 through 17, once every 4 weeks thereafter, and at Week 105 3M Environmental Laboratory Page 30 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Food Consumption Weekly for Weeks 1 through 16 and once every 4 weeks thereafter Covance 6329-212 Page 10 Clinical Pathology Frequency and Number of Animals Unscheduled Collection When possible, a blood film will be made and held for possible future examination from animalssacrificed at unscheduled intervals. Scheduled Collections Hematology, clinical chemistry, urinalysis, urine chemistry, and serum sampling will be done on 10 animals/sex/group in Groups 1 through 5 during Weeks 14,27, and 53. A blood Nm wiU be made and held for possible future examinationfor animals at scheduled sacrifices after at least 78 and 104 weeks of treatment. Method of Collection Hematology, Clinical Chemistry, Urinalyses, Urine Chemistry, Serum SampIf5 Animals will be fasted overnight;blood will be collected from a jugular vein. The anticoagulant will be potassiumEDTA for hematologytests. Samples for clinical chemistry and serum samples will be collected without anticoagulant. Urine will be collected chilled overnight (approximately 16 hours). Blood Films Blood films will be taken as part of the necropsy procedure. 3M Environmental Laboratory Page 31 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Covance 6329-212 Page 11 Tests Hematology red blood cell (erythrocyte)count hemoglobin hematocrit mean corpuscular volume mean corpuscular hemoglobin mean corpuscular hemoglobin concentration platelet count white blood cell (leukocyte) count differential blood cell count blood cell morphology reticulocyte smear (made, but not examined) glucose urea nitrogen creatinine total protein albumin globulin cholesterol total bilirubin Clinical Chemistry alanine aminotransferase gamma glutamyltransferase aspartate aminotransferase calcium inorganic phosphorus sodium potassium chloride appearance volume specific gravity PH protein urobilinogen sodium potassium Urinalysis glucose ketones bilirubin blood microscopic examination of sediment Urine Chemistry 16 hour excretion of: sodium potassium 3M Environmental Laboratory Page 32 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Covance 6329-212 Serum Samples Serum not used for clinical chemistry will be stored in a freezer set to maintain -60 to -80C.Samples will be packed on dry ice and shipped to: Kris J. Hansen, PhD 3M Environmental Technology and Safety Services 935 Bush Avenue Building 2-3E-09 St. Paul, Minnesota 55133-3331 Telephone No. : 612.778.60 18 Facsimile No.: 612.778.6176 Samples will be retained by the Sponsor for possible future analysis. Hepatocellular Proliferation and Biochemical Analyses Frequency and Number of Animals Five animals/sex in Groups 1 through 5 during Weeks 3 and 14 Cell Proliferation Tissue Collection and Immunohistochemical Evaluation At each interval, animalswill be fasted overnight, anesthetized with sodium pentobarbital, weighed, and exsanguinated. The abdominal cavity of each animal will be opened, and the liver will be removed, weighed, and representative samples of left lateral, right median, and right lateral lobes of the liver and any macroscopic lesions of the liver will be collected and preserved in zinc formalin. After fixation, each sample of liver will be embedded in paraffin, and the paraffin blocks will be shipped to: Sandra R. Eldridge, PhD Pathology Associates International 15 Woman's Mill Court, Suite I Frederick, Maryland 21701 Telephone No.: 301.663.1644, ext. 2201 Facsimile No: 301.663.8994 Proliferation cell nuclear antigen (PCNA) evaluation will be done on the samples. Results will be provided for inclusion in the final report. 3M Environmental Laboratory Page 33 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Covance 6329-2 12 Page 13 Palmitoyl-CoA Oxidase Tissue Collection and Analyses A sample (approximately500 mg) of the right lateral lobe of the liver will also be collected from each animal and flash-frozen in liquid nitrogen. The liver tissue will be stored in a freezer set to maintain -60 to -80C until analyzed by Covance for palmitoyl-CoA oxidase activity. Animal Disposition Animals will be discarded after liver collection. Termination Unscheduled Sacrifices and Deaths Necropsies will be done. Animals to be sacrificed will be anesthetized with sodium pentobarbital, weighed, and exsanguinated. A blood film will be taken as part of the necropsy procedure for sacrificed animals. Scheduled Sacrifices Interim Sacrifice After at least 78 weeks of treatment, 10 animals/sex from Groups 1,4, and 5 will be fasted overnight, anesthetized with sodium pentobarbital, weighed, exsanjpinated, and necropsied. A blood film will be taken as part of the necropsy procedure. Terminal Sacrifice After at least 104weeks of treatment, the remaining animals in Groups 1 through 5 will be fasted overnight, then anesthetized with sodium pentobarbital, weighed, exsanguinated, and necropsied. A blood film will be taken as part of the necropsy procedure. Recovery Sacrifice After at least 78 weeks of treatment and 26 weeks without treatment, the remaining animalsin Groups 6 and 7 Wiu be fasted overnight, then anesthetized with sodium pentobarbital, weighed, exsanguinated, and necropsied. A blood film will be taken as part of the necropsy procedure. 3M Environmental Laboratory Page 34 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Pos@nortem Procedures Covance 6329-212 Page 14 Necropsy The necropsy will include an examination of the external features of the carcass; all external body orifices; the abdominal, thoracic, and cranial cavities; organs; and tissues. Organ Weights At the scheduled sacrifices, the following organs (when present) will be weighed; paired organs will be weighed separately: adrenal (2) brain kidney (2) liver lung ovary (2) spleen testes thyroid (2) with parathyroid uterus with cervix Organ-to-body weight percentages and organ-to-brain weight ratios will be calculated. Bone Marrow Smear From the femur of each animal at scheduled sacrifices only; made but not examined Tissue Preservation The following tissues (when present) fiom each animal will be preserved in 10%neutral-buffered formalin: 3M Environmental Laboratory Page 35 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Covance 6329-212 adrenal (2) pancreas brain pituitary cecum prostate cervix rectum colon sahvary gland [mandibular (2)] duodenum sciatic nerve epididymis (2) seminal vesicle (2) esophagus skeletal muscle (thigh) eye (2) femur with bone marrow (articular surface Skin spinal cord (cervical, thoracic, and of the distal end) lumbar) Harderian gland spleen heart sternum with bone marrow ileum stomach jejunum testis (2) kidney (2) thymus lesions thyroid (2) with parathyroid liver trachea . lung with mainstem bronchi lymph node (mesenteric) mammary gland (females only) urinary bladder uterus vagina ovary (2) Histopathology Tissues (as appropriate) from each animal in Groups 1,5, and 7 and from each animal that dies or is sacrificed at an unscheduled interval will be embedded in paraffin, sectioned, stained with hematoxylin and eosin, and examined microscopically. Macroscopic lesions will also be examined microscopicallyfrom each animal in Groups 2,3,4, and 6. Reports One copy of each draft report will be sent to the Sponsor. The report will include the following information: 3M Environmental Laboratory Page 36 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Experimental Design and Methods Covance 6329-212 Page 16 Results dose analyses mortality clinical observations body weights body weight changes food consumption test material consumption clinical pathology results palmitoy1CoA oxidase activities macroscopic observations microscopic observations cell proliferation assessments (provided by the Sponsor's designee) Statistical Evaluation body weights body weight changes food consumption survival rates clinical pathology values palmitoyl CoA oxidase activities neoplastic and nonneoplastic lesions Statistical methods will be those presented in Attachments Nos. 1 and 2. For each sex, Groups 2 through 7 win be compared to Group 1 (Control). At the end of 1 year after issuance of the audited draft report, if no requested revisions or instructions to finalize have been communicated by the Sponsor, then the audited draft report will be considered 'final'and issued as the final report, signed by the study director, and submitted to the Sponsor. Any modifications or changes to the audited draft report requested 1 year after issuance will be performed at additionalcost to the Sponsor. Two copies of the signed final report (one unbound and one bound) will be sent to the Sponsor. 3M Environmental Laboratory Page 37 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Covance 6329-212 Record Retention AU raw data, documentation, records, protocol, specimens, and final report generated as a result of this study, including those items listed below, will be archived in the storage facilities of Covance-Madisonfor a period of 1 year following submission of the find report to the Sponsor. All raw data stored on magnetic media, the protocol and protocol amendments, study correspondence, and the original report will be retained by Covance. One year after submission of the final report, all of the aforementioned materials will be sent to the Sponsor, and a return fee will be charged. The Sponsor may elect to have the materials retained in the Covance archives for an additionalperiod of time, and Covance will charge a storage fee. If the Sponsor chooses to have Covance dispose of the materials, a disposal fee will be charged. protocol and protocol amendments dose preparation records in-life records animal receipt acclimation animal room maintenance randomizations dose administration clinical observations body weights food consumption sample collection clinical pathology records anatomical pathology records statistical analyses study correspondence tissue specimens (wet and in paraffin) blood, bone marrow,and tissue slides final report (original signed copy) 3M Environmental Laboratory Page 38 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Covance 6329-212 Page 18 The following supporting records will be retained at Covance-Madison but will not be archived with the study data. feed analysis records water analysis records animal room environment records refrigerator and freezer temperature records room temperature records for test material storage instrument calibration and maintenance records PCNA evaluation data and paraffin blocks and tissue slides for PCNA will be retained by Pathology Associates International. Serum samples sent to the Sponsor will be retained by the Sponsor 3M Environmental Laboratory Page 39 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 PROTOCOL APPROVAL Covance 6329-212 PaRe 19 W Andrew M. Seacat, PhD Study Monitor 3M Department of Toxicology Covance Laboratories Inc. 3M Environmental Laboratory Page 40 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Attachment No. 1 Covance 6329-212 Page 20 Statistical Analyses The statisticalmethods that will be used are described below. Only data collected on or after the first day of treatment will be analyzed statistically. One-way analysis of variance [ANOVA (Winer, 1971)) will be used (if applicable) to analyze body weights; body weight changes; food consumption; clinical chemistry and hematology values (except blood cell morphology); urine specific gravity, pH, and volume; urine chemistry values; palmitoy1CoA oxidase activities; organ weights; organ-to-body weight percentages; and organ-to-brain weight ratios.. Levene's test (Levene, 1960) will be done to test for variance homogeneity. In the case of heterogeneity of variance at p 5 0.05, transformationswill be used to stabilize the variance. ANOVA will be done on the homogeneous or transformed data. If the ANOVA is significant,Dunnett's t-test (Dunnett, 1964) will be used for pairwise comparisons between treated and control groups. If the ANOVA shows significancefor body weights at Week 1, one-way analysis of covariance [ANCOVA(Winer, 1971)) Win be used to a n a l p body weights, with initial body weights as the covariate, using untransformed data. If the ANCOVA is significant, least squares means t-test (Winer, 1971) Win be used for pairwise comparisons between treated and control groups. Group comparisons will be evaluated at the 5.096, two-tailed probability level. References Dunnett, C. W., "New Tables for Multiple Comparisons with a Control," Biometrics, -20:482-491 (1964). Levene, H., "Robust Tests for Equality of Variances," Contributions to Probability and Statistics, (eds.) I. OXkin et al., Ch. 25, pp. 278-292, Stanford University Press: Stanford, California (1960). Winer, B. J., Statistical Principles in Exuerimental Design, Second Ed., McGraw-Hill: New York, New York (1971b). 3M Environmental Laboratory Page 41 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Covance 6329-212 Attachment No. 2 Adjusted survival data are analyzed by the National Cancer Institute (NCI) Metable package (Thomas, 1977). The tests include: Graphical (Kaplan-Meier product-limit estimation curves), Cox-Tarone binary regression methods for trend and heterogeneity, and Gehan-Breslow nonparametric methods for trend and heterogeneity. Non-neoplastic lesions. Non-neoplastic lesions are analyzed by the Cochran-Armitage test for trend and the Fisher-Irwin exact test for heterogeneity (Thakur, 1985). Neoplastic lesions. Incidental tumors are analyzed by Dinse-Lagakos logistic prevalence methods (Dime, 1983) for trend and heterogeneity. Rapidly lethal and palpable tumors are analyzed in the same manner as survival. In the cases where the study pathologist can assign particular occult neoplastic lesions as the cause of death in the animals,such information will be taken into appropriate analysis as in the IARC document (Peto et al, 1980). References Thomas, D. G., Breslow, N., and Gart,J. J., "Trend and Homogeneity Analyses of Proportions and Life Table Data," Comput. Biomed. Res., u:373-381(1977). Thakur, A IC,Berry, K J., and MieIke, Jr., P.W.,"AFORTRAN Program for Testing Trend and Homogeneity mProportions," Comput. Progr. Biomed., B:229-233 (1985). Dinse, G. E., and Lagakos, S . W.,"Regression Analysis of Tumor Prevalence Data," J. Rov. Stat. SOC.Series C (ApDL Stat.L =:236-248 (1983). Peto, R., Pike, M. C., Day, N. E., Gray, R. G., Lee, P. N., Parish, S., Peto, J., Richards, S., and Wahrendorf, J., "Guidelines for Simple Sensitive Significance Tests for Carcinogenic EffectshLong-term Animal Experiments, in Long-term and Short-term Screening Assays for Carcinogens: A Critical Appraisal," Lyon: International Anencv for Cancer Research, pp. 311-426 (1980). 3M Environmental Laboratory Page 42 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 THE DRIELOPYENT SERWCES COUPANY PROTOCOL AMENDMENT NO. 1 Covance 6329-212 104-Week Dietary Carcinogenicity Study with Narrow Range (98.1%)N-Ethyl Peffluorooctanesulfonamido Ethanol in Rats Sponsor: Study Monitor: Testing Facility: Study Director: 3M, St. Paul, Minnesota Andrew M. Seacat, PhD Covance Laboratories Inc., Madison, Wisconsin Peter J. Thomford, PhD This amendment modifies the followingportions of the protocol: Effective January 21,1998 1 . Page 4, Animals, Strain. To correct the strain of the animals used in this study, delete the text in this section and replace with the following: CrlCD@(SD)IGSBR Effective February 6,1998 2. Page 7, Group Designations and Dietary Levels, Footnote C. To reflect the decision to delay the Week 3 collection, delete the text in this section and replace with the following: C Five animals/sex/group in Groups 1through 5 will be sacrificed during Weeks 4 and 14 for hepatocellularproliferation rate measurements and biochemical analyses (palmitoyl-CoAoxidation). 3M Environmental Laboratory Page 43 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Covance 6329-212 ProtocolAmendment No. 1 Page 2 3. Page 12, Hepatocellular Proliferation and Biochemical Analyses, Frequency and Number of Animals. To reflect the decision to delay the Week 3 collection, delete the text in this section and replace with the following: Five animals/sedgroup in Groups 1 through 5 during Weeks 4 and 14 - 4. Page 13, Effective February 17,1998 5. Page 12. To reflect the decision to collect serum samples for possible future analysis, add the following section after "ClinicalPathology". Serum Analyses Frequency and Number of Animals Five animals/sex/group in Groups 1 through 5 during Weeks 4 and 14 (animals selected for hepatocellular proliferation and biochemical analyses); five animals/sedgroup from Groups 1,4, and 5 (from animalsselected for interim sacrifice) after at least 78 weeks of treatment; and five animalslsedgroup from Groups 1through 5 at the terminal sacrifice Method of Collection Animals will be fasted overnight; blood (approximately2 mL) will be collected from a jugular vein. Samples will be collected without anticoagulant. 3M Environmental Laboratory Page 44 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Covance 6329-212 Protocol Amendment No. 1 Page 3 Sample Handling Blood samples will be allowed to clot at room temperature and centrifuged. Serum samples will be harvested and stored in a freezer set to maintain -60 to -80C. Samples will be packed on dry ice and shipped to Kris J. Hansen, PhD,3M Environmental Technology and Safety Services. Samples will be retained by the Sponsor for possible future analysis. 6. Page 13, Hepatocellular Proliferation and Biochemical Analyses. To reflect the decision to collect liver samples for possible future analyses, add the following section after "Palmitoyl-CoA Oxidase Tissue Collection and Analyses": Additional Liver Sample Collection At the collections during Weeks 4 and 14, the remaining liver will be stored in a freezer set to maintain -60 to -80C. Samples will be packed on dry ice and shipped to Kris J. Hansen, PhD,3M Environmental Technology and Safety Services. Samples will be retained by the Sponsor for possible future analysis. 7. Page 14, Postmortem Procedures. To reflect the decision to collect liver samples for possible future analyses, add the following section after "Bone Marrow Smear": Additional Liver Sample Collection A portion of the liver will be collected from five animals/sex/group from Groups 1, 4, and 5 at the interim sacrifice and from five animals/sex/group from Groups 1 through 5 at the terminal sacrifice and stored in a freezer set to maintain -60 to -80C. Samples will be packed on dry ice and shipped to Kris J. Hansen, PhD, 3M Environmental Technology and Safety Services. Samples will be retained by the Sponsor for possible future analysis. 8. Page 17, Record Retention, Paragraph 4. To indicate that frozen liver samples sent to the Sponsor will be retained by the Sponsor, delete this paragraph and replace with the following: 3M Environmental Laboratory Page 45 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Covance 6329-212 Protocol Amendment No.1 Page 4 Frozen liver samples and serum samples sent to the Sponsor will be retained by the Sponsor. Effective March 13,1998 9. Page 7, Group Designations and Dietary Levels. To reflect the decision to sacrificefive animaldsexlgroup from Groups 1 and 5 and to terminate the remaining animalsin Group 5 and 7 during Week 8, delete the text in this section and replace with the following: Number of Animals Dietary Levels Group Male Female (ppm NEtFOSE)' 1 (Control)b*c-d*e 70 70 0 2 (Low)c 60 60 3 3 (Mid)" 60 60 30 4 (Mid-High)"." 70 70 100 5 (HighPd 70 70 300 6 (Mid-High Recovery)f 40 40 100 7 (High Recovery)d 40 40 300 a T-6316 is 98.1% n-ethyl peffluorooctanesulfonamido ethanol (NEtFOSE); dose levels are expressed as ppm of NEtFOSE. b The control animals will receive the basal diet only. c Five animals/sex/groupin Groups 1 through 5 will be sacrificed during Week 4 and five animals/sex/group in Groups 1 through 4 will be sacrificed during Week 14for hepatocellularproliferationrate measurementsand biochemical analyses (palmitoyl-CoAoxidation). d Five animalskexlgroup in Groups 1 and 5 will be sacrificed during Week 8; the remaining animals in Groups 5 and 7 will be sacrificed and discarded during Week 8. e Ten animalslsexlgroup in Groups 1 and 4 will be designated as interim sacrifice animals and will be sacrificed after at least 78 weeks of treatment. f Animals in Group 6 will be treated for at least 78 weeks, then treatment will be discontinued, and the animalswill be observed for reversibihty, persistence, or delayed occurrence of toxic effects for at least 26 weeks posttreatment. During recovery, the animals will receive basal diet only. 3M Environmental Laboratory Page 46 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Covance 6329-212 Protocol Amendment No.1 Page 5 10. Page 7, Dosing Procedures, Method of Administration. To reflect the decision to terminate Groups 5 and 7 during Week 8, delete the text in this section and replace with the following: Dietary. Animals in Groups 1 through 4 will receive test diet for at least 104 weeks. Animals in Group 6 will receive test diet for 78 weeks only. Animals in Groups 5 and 7 will receive test diet for at least 7 weeks. 11. Page 9, Observation of Animals,Body Weights. To reflect.the decision to record body weights before sacrificeduring Week 8, add the following to this section: Body weights will also be recorded for all animalsin Groups 5 and 7 before sacrifice during Week 8. 12. Page 10, Clinical Pathology, Frequency and Number of Animals, Scheduled Collections. To reflect the decision to collect clinicalpathology samplesfrom animals in Groups 1 and 5 during Week 8 and to terminate Groups 5 and 7 during Week 8, delete the text in this section and replace with the following: Hematology, clinical chemistry, urinalysis, and urine chemistry will be done on five ammals/sex/groupfrom Groups 1 and 5 during Week 8. Hematology, clinical chemistry, urinalysis, urine chemistry, and serum sampling will be done on 10 animals/sex/group in Groups 1 through 4 during Weeks 14,27, and 53. A blood film will be made and held for possible future examinationfor animalsat scheduled sacrifices after at least 78 and 104 weeks of treatment. 13. Page 12, Serum Analyses, Frequency and Number of Animals. To reflect the decision to collect serum samples from animalsin Groups 1 and 5 during Week 8 and to terminate Groups 5 and 7 during Week 8, delete the text in this section and replace with the following: 3M Environmental Laboratory Page 47 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Covance 6329-212 Protocol Amendment No. 1 Five animals/sex/group in Groups 1 through 5 during Week 4 (animalsselected for hepatocellular proliferation and biochemical analyses); five animals/sex/group in Groups 1 and 5 during Week 8; five animals/sex/group in Groups 1 through 4 during Week 14 (animals selected for hepatocellular proliferation and biochemical analyses); five animalshedgroup from Groups 1 and 4 (from animals selected for interim sacrifice) after at least 78 weeks of treatment; and five animals/sex/group from Groups 1 through 4 at the terminal sacrifke 14. Page 12, Hepatocellular Proliferation and Biochemical Analyses, Frequency and Number of Animals. To reflect the decision to perform analyses on livers from animalsin Groups 1 and 5 during Week 8 and to terminate Groups 5 and 7 during Week 8, delete the text in this section and replace with the following: Five animals/sex/group in Groups 1 through 5 during Week 4; five animals/sex/group in Groups 1 and 5 during Week 8; and five animals/sex/group in Groups 1 through 4 during Week 14 15. Page 13, HepatocellularProliferation and Biochemical Analyses, Additional Liver Sample Collection. To reflect the decision to collect livers from animalsin Groups 1 and 5 during Week 8, delete the text in this section and replace with the following: At the collections during Weeks 4,8, and 14, a portion of the remaining liver will be stored in a freezer set to maintain -60 to -80C. Samples will be packed on dry ice and shipped to Kris J. Hansen, PhD,3M EnvironmentalTechnology and Safety Services. Samples will be retained by the Sponsor for possible future analysis. 16. Page 12, Hepatocellular Proliferation and Biochemical Analyses, Cell Proliferation Tissue Collection and Immunohistochemical Evaluation, Paragraph 1. To reflect the decision to collect only the left lateral lobe of the liver at the Week 8 and 14 collections, delete the text in this paragraph and replace with the following: 3M Environmental Laboratory Page 48 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Covance 6329-212 Protocol Amendment No.1 Page 7 At each interval, animalswill be fasted overnight, anesthetized with sodium pentobarbital, weighed, and exsanguinated. The abdominalcavity of each animal will be opened, and the liver will be removed and weighed. At the Week 4 collection, representative samples of left lateral, right median, and right lateral lobes of the liver and any macroscopic lesions of the liver will be collected and preserved in zinc formalin. At the Week 8 and 14 collections, representative samples of the left lateral lobe of the liver and any macroscopic lesions of the liver will be collected and preserved in zinc formalin. 17. Page 13, Termination, Scheduled Sacrifices. To reflect the decision to sacrifice five animals/sex/group from Groups 1 and 5,to terminate the remaining animalsin Groups 5 and 7 during Week 8, and to sacrifice five animals/sex/group from Groups 1 through 4, delete the text in this section and replace with the following: Interim Sacrifices During Week 8, five animals/sex/groupfrom Groups 1 and 5 will be fasted overnight, bled for clinical pathology and serum analyses, anesthetized with sodium pentobarbital, weighed, exsanguinated, and necropsied. Liver samples will be collected for palimitoyl CoA, PCNA, and frozen samples for possible analysis. The remaining animalsin Groups 5 and 7 will be euthanatized with carbon dioxide and discarded without necropsy. During Week 14, five animals/sex/group from Groups 1 through 4 will be fasted overnight, bled for clinical pathology and serum analyses, anesthetized with carbon dioxide, weighed, exsanguinated, and necropsied. Liver samples will be collected for palimitoyl CoA, PCNA, and frozen samples for possible analysis. After at least 78 weeks of treatment, five animals/sex/groupfrom Group 1 and 10 animals/sex from Group 4 will be fasted overnight, anesthetized with carbon dioxide, weighed, exsanguinated, and necropsied. A blood film will be taken as part of the necropsy procedure. 3M Environmental Laboratory Page 49 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Covance 6329-212 Protocol Amendment No. 1 Page 8 .Terminal Sacrifice After at least 104 weeks of treatment, the remaining animalsin Groups 1through 4 will be fasted overnight, then anesthetized with carbon dioxide, weighed, exsanguinated, and necropsied. A blood film will be taken as part of the necropsy procedure. Recovery Sacrifice After at least 78 weeks of treatment and 26 weeks without treatment, the remaining animalsin Group 6 will be fasted overnight, then anesthetized with carbon dioxide, weighed, exsanguinated, and necropsied. A blood film will be taken as part of the necropsy procedure. 18. Page 14, Postmortem Procedures, Additional Liver Sample Collection. To reflect the decision to sacrifice five animals/sex/group from Groups 1and 5 and to terminate the remaining animalsin Group 5 and 7 during Week 8, delete the text in this section and replace with the following: A portion of the liver will be collected from five animals/sex/group from Groups 1 and 4 at the Week 79 sacrifice and from five animals/sex/group from Groups 1 through 4 at the terminal sacrifice and stored in a freezer set to maintain -60 to -80C. Samples will be packed on dry ice and shipped to Kris J. Hansen, PhD,3M Environmental Technology and Safety Services. Samples will be retained by the Sponsor for possible future analysis. 19. Page 15, Postmortem Procedures, Histopathology. To reflect the decision to terminate Groups 5 and 7 early, to examine tissues from animalssacrificed during Weeks 8 and 14, and to examine tissues from Groups 4 and 6, delete the text in this section and replace with the following: Tissues (as appropriate) from each animal in Groups 1,4, and 6 sacrificed at the Week 79 interim sacrifice and terminal sacrifice and from each animal that dies or is sacrificed at an unscheduled interval will be embedded in paraffin, sectioned, stained with hematoxylin and eosin, and examined microscopically. Adrenals, brain, eyes, kidneys, liver, mesenteric lymph node, pancreas, spleen, testes, and 3M Environmental Laboratory Page 50 3M Medical Department Study: T6316.1 .. Analytical Report: FACT-TOX-001 LRN-U2103 Covance 6329-212 Protocol AmendmentNo.1 Page 9 ,ovariesfrom the animals necropsied at Weeks 8 and 14 interim sacrifice will also be embedded in paraffin, sectioned, stained with hematoxylin and eosin, and examined microscopically. Macroscopic lesions will also be examined microscopically from each animalin Groups 2 and 3 sacrificed at terminal necropsy. 20. Page 16, Reports, Results. To reflect the decision to include results from Covance 6329-228 as an appendix to this report, add the following: The study report for Covance 6329-228 will be included as an appendix to this report. Effective March 30,1998 21. Page 12, Hepatocellular Proliferation and Biochemical Analyses, Cell Proliferation Tissue Collection and Immunohistochemical Evaluation, Paragraph 3. To include the microscopic examinations of liver sections stained with hematoxylin and eosin and to indicate that evaluations will be done on the left lateral lobe of the liver only, delete the text in this section and replace with the following: Proliferation cell nuclear antigen (PCNA) evaluation will be done on the samples (left lateral lobe only). In addition, sections of the left lateral lobe of the liver will be stained with hematoxylin and eosin and examined microscopically. Results will be provided for inclusion in the final report. 3M Environmental Laboratory Page 51 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 AMENDMENT APPROVAL Covance 6329-212 Protocol Amendment No. 1 Page 10 Study Monitor 3M Covance Laboratories Inc. Date , ~ 3M Environmental Laboratory Page 52 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 c o v m TMEDEVELOPMENTSERYlCESWYPANY PROTOCOL AMENDMENT NO.2 Covance 6329-212 104Week Dietary Carcinogenicity Study with Narrow Range (98.1%) N-Ethyl Perfluorooctanesulfonamido Ethanol in Rats Sponsor: Study Monitor: Testing Facility: Studv Director: 3M, St. Paul, Minnesota Andrew M. Seacat, PhD Covance Laboratories Inc., Madison, Wisconsin Peter J. Thomford, PhD This amendment modifies the following portions of the protocol: Effective April 23,1998 1. Page 13, Termination, Unscheduled Sacrificesand Deaths. To reflect the decision to anesthetize animals with carbon dioxide for unscheduled sacrifices, delete the text in this section and replace with the following: Necropsies will be done. Animals to be sacrificed will be anesthetized, weighed, and exsanguinated. Animals sacrificed before Week 13 will be anesthetized with sodium pentobarbital, animals sacrificed during or after Week 13 will be anesthetized with carbon dioxide. A blood film will be taken as part of the necropsy procedure for sacrificed animals. Effective April 28,1998 2. Page 2, Alternate Study Monitor. To include alternate study monitor in the protocol, add the following after "Study Monitor:" 3M Environmental Laboratory Page 53 .3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Alternate Study Monitor Marvin T. Case, DVM, PhD 3M Toxicology Services Telephone No. : 612.733.5 180 Facsimile No.: 612.733.1773 Covance 6329-212 Protocol Amendment No. 2 Page 2 3. Page 7, Group Designations and Dietary Levels. To reflect the decision to begin recovery after 52 weeks of treatment and to change the Week 79 interim sacrificeto Week 53, delete the text in this section and replace with the following: Group Number of Animals Male Female Dietary Levels (ppm NEtFOSE)" 1 (Control)b*c*4e 70 2 (Low>" 60 3 (Mid)" 60 4 (Mid-High)"." 70 5 (HighYd 70 6 (Mid-High Recovery)' 40 7 Wgh Recovery)d 40 70 0 60 3 60 30 70 100 70 --3w 40 100 40 - 300 a T-6316 is 98.1% n-ethyl perfluorooctanesulfonamido ethanol (NEtFOSE); dose levels are expressed as ppm of NEtFOSE. b The control animalswill receive the basal diet only. c Five animalslsexlgroupin Groups 1through 5 will be sacrificed during Week 4, and five animals/sex/groupin Groups 1 through 4 will be sacrificed during Week 14 for hepatocellularproliferation rate measurements and biochemical analyses (palmitoyl-CoAoxidation). d Five animals/sex/group in Groups 1 and 5 will be sacrificed during Week 8; the remaining animalsin Groups 5 and 7 will be sacrificed and discarded during Week 8. e Ten animals/sex/group in Groups 1 and 4 will be designated as interim sacrifice animals and will be sacrificed after at least 52 weeks of treatment. f Animals in Group 6 will be treated for at least 52 weeks, then treatment will be discontinued, and the animalswill be observed for reversibility, persistence, or delayed occurrence of toxic effects for at least 52 weeks posttreatment. During recovery, the animals will receive basal diet only. 3M Environmental Laboratory Page 54 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Covance 6329-212 Protocol Amendment No. 2 4. Page 7, Dosing Procedures, Method of Administration. To reflect the decision to beg& recovery after 52 weeks of treatment, delete the text in this section and replace with the following: Dietary. Animalsin Groups 1 through 4 wdl receive test diet for at least 104weeks. Animals in Group 6 will receive test diet for 52 weeks only. Animals in Groups 5 and 7 will receive test diet for at least 7 weeks. 5. Page 10, Clinical Pathology, Frequency and Number of Animals,Scheduled Collections, Paragraph 2. The Week 79 interim sacrifice will be moved to Week 53, therefore blood films will be prepared from animalssacrificed after 52 weeks of treatment. To reflect this decision, delete the text in this paragraph and replace with the following: A blood film will be made and held for possible future examinationfor animals at scheduled sacrifces after at least 52 and 104 weeks of treatment. 6. Page 12, Serum Analyses, Frequency and Number of Animals. The Week 79 interim sacrifice will be moved to Week 53, therefore serum samples will be collected from animals sacrificed after 52 weeks of treatment. To reflect this decision, delete the text in this section and replace with the following: Five animals/sex/groupin Groups 1 through 5 during Week 4 (animalsselected for hepatocellular proliferation and biochemical analyses); five animals/sex/group in Groups 1 and 5 during Week 8; five animals/sex/group in Groups 1 through 4 during Week 14 (animalsselected for hepatocellular proliferation and biochemical analyses); five animals/sex/group from Groups 1 and 4 (from animals selected for interim sacrifice) after at least 52 weeks of treatment; and five anjmals/sex/group from Groups 1 through 4 at the terminal sacrifice. 7. Page 13, Termination, Scheduled Sacrifices. To reflect the decision to move the Week 79 interim sacrifice to Week 53 and to begin recovery after 52 weeks of treatment, delete the text in this section and replace with the following: 3M Environmental Laboratory Page 55 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Covance 6329-212 Protocol Amendment No.2 Interim Sacrifices During Week 8, five animalshedgroup from Groups 1 and 5 will be fasted overnight,bled for clinicalpathology and serum analyses, anesthetized with sodium pentobarbital, weighed, exsanguinated,and necropsied. Liver samples will be collected for palimitoyl CoA analysis and PCNA evaluation; additional liver samples will be frozen for possible analysis. The remaining animals in Groups 5 and 7 will be euthanatized with carbon dioxide and discarded without necropsy. During Week 14, five animaldsedgroup from Groups 1through 4 will be fasted overnight, bled for clinicalpathology and serum analyses, anesthetized with carbon dioxide, weighed, exsanguinated,and necropsied. Liver samples will be collected for palimitoyl CoA analysis and PCNA evaluation; additional liver samples will be frozen for possible analysis for possible analysis. After at least 52 weeks of treatment, five animals/sedgroup from Group 1 and 10 animals/sex from Group 4 will be fasted overnight, anesthetized with carbon dioxide, weighed, exsanguinated, and necropsied. Terminal Sacrifice After at least 104 weeks of treatment,the remaining animalsin Groups 1 through 4 will be fasted overnight, then anesthetizedwith carbon dioxide, weighed, exsanguinated,and necropsied. A blood film will be taken as part ofthe necropsy procedure. Recovery Sacrifice M e r at least 52 weeks of treatment and 52 weeks without treatment, the remaining animalsin Group 6 will be fasted overnight, then anesthetized with carbon dioxide, weighed, exsanguinated, and necropsied. A blood film will be taken as part of the necropsy procedure. 8. Page 14, Postmortem Procedures, Organ Weights. To reflect the decision to record organ weights at the Week 8, 14, and 53 interim sacrifices only, delete the text in thrs section and replace with the following: 3M Environmental Laboratory Page 56 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Covance 6329-212 Protocol Amendment No. 2 At the Week 8, 14, and 53 interim sacrifices, the following organs (when present) will be weighed; paired organs will be weighed separately: adrenal (2) brain kidney (2) liver lung ovary (2) spleen testis (2) thyroid (2) with parathyroid uterus with cervjx Organ-to-body weight percentages and organ-to-brainweight ratios will be calculated. 9. Page 14, Postmortem Procedures, Additional Liver Sample Collection. The Week 79 interim sacrifice will be moved to Week 53,therefore liver samples will be collected from animals sacrificed after 52 weeks of treatment. To reflect this decision, delete the text in this section and replace with the following: A portion of the liver will be collected from five animals/sex/group from Groups 1 and 4 at the Week 53 sacrifice and from five animals/sex/group from Groups 1 through 4 at the terminal sacrifice and stored in a freezer set to maintain -60 to -80C. Samples will be packed on dry ice and shipped to Kris J. Hansen, PhD, 3M Environmental Technology and Safety Services. Samples will be retained by the Sponsor for possible future analysis. 10. Page 15, Postmortem Procedures, Histopathology, Paragraph 1. The Week 79 interim sacrifice will be moved to Week 53, therefore histopathology will be done for animals sacrificed after 52 weeks of treatment. To reflect this decision, delete the text in this section and replace with the following: 3M Environmental Laboratory Page 57 3M Medical Department Study: T6316.1 i' c . Analytical Report: FACT-TOX-001 LRN-U2103 Covance 6329-212 Protocol Amendment No.2 Tissues (as appropriate) from each animal in Groups 1,4, and 6 sacrificed at the Week 53 interim sacrifice and terminal sacrifice and from each animal that dies or is sacrificed at an unscheduled interval will be embedded in parain, sectioned, stained with hematoxylin and eosin, and examined microscopically. Adrenals, brain, eyes, kidneys, liver, mesenteric lymph node, pancreas, spleen, testes, and ovaries from the animalsnecropsied at the Week 8 and 14 interim sacrifices will also be embedded in paraffin, sectioned, stained with hematoxylin and eosin, and examined microscopically. 11. Page 15, Reports, Paragraph 1. To reflect the decision to provide unaudited summary reports after the Week 14 and 53 interim sacrifices, delete the text inthis paragraph and replace with the following: After the Week 14 and 53 interim sacrifices, unaudited summary reports will be sent to the Sponsor. The summary reports will include a brief description of methods and results and summary tables of in-life data, clinical pathology data, and anatomical pathology data. After completion of the study, one copy of the draft report will be sent to the Sponsor. The draft report will include the following information: Effective May 15,1998 12. Page 12, Serum Analyses, Method of Collection. To reflect the decision to increase the volume collected for serum analyses at the Week 53 and 105 sacrifices, delete the text in this section and replace with following: Animals will be fasted overnight; blood (approximately2 mL at the Week 4, 8 and 14 sacrifices, approximately 3 mL at the Week 53 and 105 sacrifices) will be collected from a jugular vein. Samples wdl be collected without anticoagulant. 3M Environmental Laboratory Page 58 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 AMENDMENT APPROVAL C O V ~ I6I C32~9-212 Protocol Amendment No.2 Page 7 Andrew M. Seacat, PhD Study Monitor 3M Covhce Laboratories Inc. 3M Environmental Laboratory Page 59 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 covm PROTOCOL AMENDMENT NO. 3 Covance 6329-212 1WWeek Dietary CarcinogenicityStudy with Narrow Range (98.1%) N-Ethyl PeduorooctanesulfonamidoEthanol m Rats sponsor: Study Monitor: Testing Facility: Study Director: 3M,St. Paul, Minnesota Andrew M. Seacat, PhD Covance LaboratoriesInc., Madison, Wisconsin Peter J. Thomford, PhD This amendment modifies the following portions of the protocol: Effective January 21,1998 1. Page 4. To include the vehicle used to dissolve the test material before mixing with the diet, add the following section. Vehicle Identification Acetone Lot Numbers The lot numbers will be maintained in the raw data. MtY On file with the manufacturer Stability On file with the manufacturer Storage Conditions At room temperature 3M Environmental Laboratory Page 60 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Covance 6329-212 Protocol Amendment No. 3 characteristics Information on synthesismethods, composition, or other characteristics that define the vehicle is on file with the manufacturer. 2. Page 16, Reports, Results. To include organ weights as a result to be reported, add "organ weights" to this section. 3. Page 16, Reports, Statistical Evaluation. To include statistical evaluation of organs weights, add "organ weights (Weeks 8, 14, and 53 interim sacrifices only)" Effective April 28,1998 4. Page 7, Group Designationsand Dietary Levels, Footnote e. To correct the number of Group 1animalsto be sacrificed at the Week 53 interim sacrifice, delete thissentence and replace with the foIIowing: e Five aninddsex in Group 1 and 10 animals/sex m Group 4 will be designated as interim sacrifice animals and will be s a c s c e d after at least 52 weeks of treatment. Effective June 30,1998 5. Page 8, Dosing Procedure, Retention Samples. To reflect the decision to send retention samples of control diet to the Sponsor, delete the text m this section and replace with the following: Samples (approximately 100 g) Win be taken from each dose preparation during the in-life phase and stored at room temperature. Unless used for analyses, retention samples from the treated groups Win be discarded at least 1 month after completion of the in-life phase. Samples from control diets prepared for Weeks 24 and before will be shipped to: 3M Environmental Laboratory Page 61 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Covance 6329212 Protocol Amendment No.3 Kris J. Hartsen, PhD 3M EnvironmentalTechnology and Safety Services 935Bush Avenue Building 2-3E-09 St. Paul, Minnesota 55133-3331 Telephone No.: 651.778.6018 Facsimile No.: 651.778.6176 Efkctive July 20,1998 6. Page 2, Study Monitor and Alternate Study Monitor. To indicate the change in the area code, delete the text in these sections and replace with the following: Study Monitor Andrew M.Seacat, PhD 3M Telephone No.: 651.575.3161 Facsimile No.: 651,733,1773 Alternate Study Monitor Marvin T. Case, DVM,PhD 3M Toxicology Services Telephone No.: 651.733.5180 Facsimile No.: 651.733.1773 7. Page 4, Disposition of Test Material. To indicate the change m the area code, delete the text m these section and replace with the following: After authorization fiom the Sponsor, any remaining test material will be returned to: Andrew M. Seacat, PhD 3M Toxicology Services Building 220-2E-O2,3MCenter St. Paul, Minnesota 55144-1000 Telephone No.: 651-575.3161 Facsimile No.: 651.733.1733 3M Environmental Laboratory Page 62 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Covance 6329212 Protocol Amendment No. 3 8. Page 12, Clinical Pathology, Serum Samples. To indicate the change in the area code, delete the text m these section and replace with the following: Serum not used for clinical chemistry will be stored in a freezer set to maintain -60 to -80C. Samples will be packed on dry ice and shipped to: Kris J. Hamen, PhD 3M Environmental Technology and Safety Services 935 Bush Avenue Building 2-3E-09 St. Paul, Minnesota 55133-3331 Telephone No.: 651.778.6018 Facsimile No.: 651.778.6176 Samples will be retained by the Sponsor for possible future analysis. 3M Environmental Laboratory Page 63 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 A M E N D ~ N TAPPROVAL Covance 6329212 Protocol Amendmemt No. 3 Andrew M. Seacat, PhD Study Monitor 3M Study Direztor Covance Laboratories Inc. Date ' 3M Environmental Laboratory Page 64 3M Medical Department Study: T6316.1 c o v m TnEDEVELOPYEN7SERVKIS COYPANV Analytical Report: FACT-TOX-001 LRN-U2103 PROTOCOL AMENDMENT NO.4 Covance 6329-212 104-Week Dietary CarcinogenicityStudy with Narrow Range (98.1%)N-Ethyl Perfluorooctanesulfonamido Ethanol in Rats Sponsor: Study Monitor: Testing Facility: Study Director: 3M,St.Paul, Minnesota Andrew M. Seacat, PhD Covance LaboratoriesInc., Madison, Wisconsin Peter J. Thomford, PhD This amendment modifies the following portion of the protocol. Effective January 13,2000 1. Page 14, Postmortem Procedures, Tissue Preservation. The Sponsor has requested that a subset of tissuesbe collected for possible electron microscopy. Therefore, add the following to thissection At the terminalsacrifice, sections of the heart and liver will be collected from 10 animals/sex/group m Groups 1,3,4, and 6 and preserved m 2.0% paraformaldehyde/2.5% glutaraldehyde in 0.1 M phosphate buffer. These tissues win be processed and embedded in epoxy blocks for possible future examination by electron microscopy. 3M Environmental Laboratory Page 65 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 AMENDMEN" APPROVAL Covance 6329-2 12 Protocol Amendment No.4 Page 2 &&?if Andrew M.Seacat, PhD Study Monitor 3M ~ovanceLaboratories ~nc: 3M Environmental Laboratory Page 66 3M Medical Department Study: T6316.1 ? c o v m WE DEVELOPMENTSERYKXS COMPANY Analytical Report: FACT-TOX-001 LRN-U2103 PROTOCOL AMENDMENT NO.5 Covance 6329-212 104-Week Dietary Carcinogenicity Study with Narrow Range (98.1%) N-Et,.yl Perfluorooctanesulfonamido Ethanol in Rats Sponsor: Study Monitor: Testing Facility: Study Director: 3M, St. Paul, Minnesota Andrew M. Seacat, PhD Covance Laboratories Inc., Madison, Wisconsin. Peter J. Thomford, PhD This amendment modifies the following portion of the protocol. EffectiveJanuary 21,2000 1. Page 10, Clinical Pathology, Frequency and Number of Animals,Scheduled Collections. The Sponsor has requested that specific clinical chemistry parameters be determined for animals at terminaland recovery sacrifice. To & e c t this decision, add the following to thissection In addition, samples for clinicalchemistryparameters (cholesteroland triglycerides) will be collected from all remaining animals at terminal and recovery sacrifice. 2. Page 12, Serum Analyses, Frequency and Number of Animals. The Sponsor has requested that serum be collected from all animalsat terminal and recovery sacrifice. To reflect this decision, delete the text in this section and replace with the following. Five animals/sex/group in Groups I through 5 during Week 4 (animalsselected for hepatocellular proliferation and biochemical analyses); five animals/sex/group in Groups 1 and 5 during Week 8; five animals/sex/group in Groups 1through 4 during Week 14 (animalsselected for hepatocellular proliferation and biochemical analyses); five anirnals/sex/groupfrom Groups 1 and 4 (from animals selected for interim sacrifice) after at least 52 weeks of treatment; and all remaining animals at terminal and recovery sacrifice. 3M Environmental Laboratory Page 67 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Covance 6329-212 Protocol Amendment No.5 3. Page 14, Postmortem Procedures,Additional Liver Sample Collection. The Sponsor has requested that frozen liver samples be collected from all animalsat terminal and recovery sacrifice. Therefore, delete the text in this section and replace with the following. A portion of the liver will be collected from five animals/sex/group from Groups 1 and 4 at theWeek 53 sacrifice and from all remaining animalsat the terminal and recovery sacrifice and stored in a freezer set to maintain -60 to -80C. Samples will be packed on dry ice and shipped to Kris J. Hansen, PhD, 3M Environmental Technology and Safety Services. Samples will be retained by the Sponsor for possible future analysis. EffectiveJanuary 26,2000 4 Page 15, Postmortem Procedures, Histopathology. The Sponsor has requested that all tissues from animalsin Groups 1and 4 at terminal sacrifice and selected tissues from animals in Groups 2,3, and 6 at the terminal and recovery sacrifice be examined microscopically. To reflect this decision, delete the text in thissection and replace with the following. Tissues (as appropriate) from each animalin Groups 1and 4 sacrificed at the terminal sacrifice and from each animalthat dies or is sacrificed at an unscheduled interval wiIl be embedded in paraffin,sectioned, stained with hematoxylin and eosin, and examined microscopically. Tissues (as appropriate) fiom each animal in Groups 1,4, and 6 sacrificed at the Week 53 interim sacrifice will be embedded in paraffin, sectioned, stained with hematoxylin and eosin, and examined microscopically. Lesions, liver, lungs, kidneys, pancreas, thyroid, testes, and mammary glands (females) from each animalin Groups 2,3, and 6 sacrificed at the terminal and recovery sacrifice wiU be embedded in paraffin, sectioned, stained with hematoxylin and eosin, and examined microscopically. Parathyroids will be processed with the thyroids, but will not be examined. Adrenals, brain, eyes, kidneys, liver, mesenteric lymph node, pancreas, spleen, testes, and ovaries fromthe animalsnecropsied at the Week 8 and 14 interim sacrifices will also be embedded in paraffin, sectioned, stained with hematoxylin and eosin, and examined microscopically. 3M Environmental Laboratory Page 68 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 AMENDMENT APPROVAL Covance 6329-212 Protocol Amendment No.5 Page 3 - Andrew M. Seacat, PhD Study Monitor 3M 3M Environmental Laboratory Page 69 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 3M Environmental Laboratory Protocol - Analytical Study Phase 104-Week Dietary Carcinogenicity Study with Narrow Range N-Ethyl PerfluorooctanesulfonamidoEthanol in Rats In-Vivo Study Reference Number: Covance # 6329-212 Study Number: AMDT-092597.1 Test Material: T-6316 Name and Address of Sponsor: 3M Toxicology Services Building 220-2E-O2,3M Center St. Paul, MN 55144-1000 Name and Address of Testing Facility: 3M Environmental Technology and Services 935 Bush Avenue St. Paul, MN 55106 Proposed Initiation Date: April 7,1998 Proposed Completion Date: October, 31,1998 Method Numbers and Revisions FACT-M-1.0, Extraction of Perfluorooctanesulfonate from Liver for Analysis using HPLC-ElectrosprayNass Spectrometry FACT-M-2.0, Analysis of Liver Extracts for Fluorochemicals using HPLC- ElectrosprayMass Spectrometry FACT-M-3.0, Extraction ofPerfluorooctanesulfonatefrom Sera for Analysis using HPLC-ElectrosprayNas Spectrometry FACT-M-4.0, Analysis of Sera Extracts for Fluorochemicals using HPLC- ElectrosprayMass Spectrometry Author: Lisa Clemen Kris J. Handen, PhD Study Director II ?[ f t Date Dale Bacon Date Study Director Management Andrew M. Seacat, PG Dhe Sponsor Representative 3M Environmental Laboratory Page 1 of 5 Page 70 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 1.0 PURPOSE 1.1 According to this analytical protocol, the 3M Environmental Laboratory will analyze the tissue and fluid samples fiom the Covance study number 6329-212, "104-Week Dietary Carcinogenicity with Narrow Range N-Ethyl Perfluorooctanesulfonamido Ethanol in Rats." The collected data will be given to the sponsor for their use in the assessment of toxicological effects of the test material when administered in the diets of rats for at least 104 weeks. 1.2 Data collected in the Environmental Laboratory will be considered non-quantitative screening data until future studies have been conducted to determine absolute recoveries of specific or general fluorochemical compounds. 2.0 REGULATORY COMPLIANCE 2.1 This analytical phase of the study will be conducted in accordance with the FDA Good Laboratory Practices Regulations 21 CFR 58, with the following exceptions: 2.1.1 The analytical phase is being conducted as a separate study and therefore has a separate Study Director, protocol, and f m d report, from those listed in the Covance protocol 6329-212. 2.1.2 The characterization of the reference material, including purity, identity, and stability, are the responsibility of the sponsor. 2.1.3 Sample storage stability will not be determined. 3.0 TEST MATERIALS 3.1 Control, and reference Materials and Matrices 3.1.1 Analytical Reference Material: T-6316, from 3M ICPRCP Division 3.1.2 Analytical Reference Matrix: Rat liver, from Covance and rat serum, from Sigma Chemical Company. 3.1.3 Analytical Control Material: None. 3.1.4 Analytical Control Matrix: Rat liver, fiom Covance and rat serum, fkom Sigma Chemical Company. 3.2 Number of Test and Control Samples: Liver and serum fiom 680 test animals and 140 control animals will be made available; samples will be analyzed as requested by the sponsor or the study director. Other biological tissues (kidney, bile, dermal application site, and cellular fraction) will be available for analysis if deemed appropriate. 3.3 Identification of Test and Control Samples: The samples will be identified using the Covance animal identification number which consists of a letter and five digit number, plus the tissue identity and day identity (serum). 3.4 Purity and Identity of Reference Material: To be determined by Sponsor. 3.5 Stability of Reference Material: To be determined by Sponsor. 3.6 Storage Conditions for Reference Materials: Reference materials will be stored at room temperature (3.1.l), and samples will be stored at -20 f 10C (3.1.2,3.1.4). Test and Control samples will be received according to AMDT-S-10-0. 3M Environmental Laboratory Page 2 of 5 Page 71 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 3.7 Disposition of Specimens: Biological tissues and fluids will be retained per GLP Regulation for the time period required for studies longer than 28 days. 3.8 Safety Precautions: Refer to appropriate MSDS. Wear appropriate laboratory attire. Use caution when handling knives for cutting tissue samples. 4.0 EXPERIMENTAL - Overview 4.1 The tissues from animals dosed as described (Covance# 6329-212), will be available for analysis for fluorine-containingcompounds. At the discretionof the Study Director, a series of analytical tests can be performed. All high dose and control sera and livers will be analyzed initially using HPLC-electrospray mass spectrometry to identify fluorine-containing compounds of interest present in the sera and liver (if any). The screening for fluoride in liver via combustion may be performed to present definitive data for fluorine in the liver. Based on the findings from these analyses, additional samples, tissues, or fluids may be analyzed at the discretion of the Study Director to determine the presence of fluorochemicals in these matrices. 5.0 EXPENMENTAL - Methods 5.1 Methods (attached): 5.1.1 FACT-M-1.0, "Extraction of Perfluorooctanesulfonatefrom Liver for Analysis using HPLC-ElectrosprayMass Spectrometry'' 5.1.2 FACT-M-2.0, "Analysis of Liver Extracts for Fluorochemicals using HPLCElectrosprayMass Spectrometry'' 5.1.3 FACT-M-3.0, "Extraction of Perfluorooctanesulfonate from Serum for Analysis using HPLC-ElectrosprayM a s s Spectrometry" 5.1.4 FACT-M-4.0, "Analysis of Serum Extracts for Fluorochemicals using HPLCElectrosprayMass Spectrometry'' 6.0 DATA ANALYSIS 6.1 Quality Control: Matrix spikes will be extracted and analyzed to determine accuracy of the method. Also, continuing calibration checks will be analyzed to determine response bias. 6.2 Transformations: Any transformationsperformed on data collected during the analytical phase of the study will be documented in the final report. 6.3 Statistics: At the discretion of the Study Director, statistics used may include regression analysis of serum concentrationswith time, averages, and standard deviationsof concentrationsfor the different dose groups. If necessary, simple tests such as the Student's t-test may be applied to determine statistical difference. Any statistical analysis performed will be documented in the final report. 6.4 Data Reporting: A final data package will be submitted to 3M Toxicology Services. The data package will include the following with additional data included as deemed appropriate. 6.4.1 A summary of individual sample results, reported as a concentration (weightlweight, weightlvolume) of fluoride per tissue or fluid, or as the mass of a specific fluorochemical (HPLC-electrospray mass spectrometry) per unit of tissue or fluid. 6.4.2 A summary of quality control results (continuing calibration checks, method blanks, instrument blanks, matrix spikes, and matrix spike duplicates). 3M Environmental Laboratory Page 3 of 5 Page 72 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 6.4.3 Certified copies or originals of the written validated methods. 6.4.4 Certified copies or originals of sample identification sheets sent from Covance. 6.4.5 Certified copies or originals of study specific raw data. 6.4.6 A summary of key personnel involved with the analytical phase of the study. 6.4.7 A signed QAU statement listing the dates of inspections and reports of findings to management and Study Director. 7.0 MAINTENANCE OF RAW DATA AND RECORDS 7.1 The following raw data and records (or certified copies thereof) will be maintained in the study folder in the archives according to appropriate SOPs. 7.1.1 7.1.2 7.1.3 7.1.4 7.1.5 7.1.6 7.1.7 Approved protocol Approved methods Data summaries Study correspondence Shipping records Raw data Electronic copies of data 7.2 Supporting records to be retained separately from the study folder in the archives according to 3M ET & S S SOPs, will include, but not necessarily be limited to the following: 7.2.1 7.2.2 7.2.3 7.2.4 7.2.5 7.2.6 Approved validation reports Training records Calibration records Instrument maintenance logs Standard operating procedures, equipment procedures, and methods Appropriate specimens 8.0 REFERENCES 8.1 AMDT-S-1, "Chemical Tracking" 8.2 AMDT-S-2, "Solutions and Standards Making" 8.3 AMDT-S-3, "Training" 8.4 AMDT-S-4, "General Lab Documentation Systems" 8.5 AMDT-S-5, "GLP-related Documentation Systems" 8.6 AMDT-S-6, "General Lab Records" 8.7 AMDT-S-7, "GLP-Related Records" 8.8 AMDT-S-8, "Archives" 8.9 AMDT-S-9, "GLP Program and Responsibilities" 8.10 AMDT-S-10, "Sample Tracking System" 3M Environmental Laboratory Page 4 of 5 Page 73 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 8.11 AMDT-S-12, "Analytical Method Validation" 8.12 AMDT-S-13, "Equipment Installation" 8.13 AMDT-S-14, "Quality Assurance Unit" 8.14 AMDT-S-15, "Laboratory Practices and Data Management" 8.15 AMDT-S-17, "Analytical Equipment Systems Documentation" 8.16 AMDT-S-18, "Routine Calibration Checks of Balances" 8.17 AMDT-S-20, "Daily Calibration Check of Automatic Pipettors" 8.18 AMDT-S-21, "Annual Calibration Checks of Automatic Pipettors" 8.19 AMDT-S-25, "Labeling, Storage, and Use of Test, Control, and Reference Substances" 8.20 AMDT-S-30, "Project Control and Data Review in the AMDT" 9.0 ATTACHMENTS 9.1 FACT-M-1.0, Extraction of Perfluorooctanesulfonate fiom Liver for Analysis using HPLC- ElectrosprayMass Spectrometry 9.2 FACT-M-2.0, Analysis of Liver Extracts for Fluorochemicals using HPLC- ElectrosprayMass Spectrometry 9.3 FACT-M-3.0, Extraction of Perfluorooctanesulfonate from Serum for Analysis using HPLC- ElectrosprayMass Spectrometry 9.4 FACT-M-4.0, Analysis of Serum Extracts for Fluorochemicals using HPLC- ElectrosprayMass Spectrometry 3M Environmental Laboratory Page 5 of 5 Page 74 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Study Title 104-Week Dietary Carcinogenicity Study with Narrow Range N-Ethyl PerfluorooctanesulfonamidoEthanol in Rats PROTOCOL AMENDMENT NO. 1 Amendment Date: 20 January 2000 Performing Laboratory 3M Environmental Technology & Safety Services 3M Environmental Laboratory 935 Bush Avenue St. Paul, MN 55106 Laboratory Project Identification ET&SS LRN-U2103 FACT TOX-001 Covance Study: 6329-212 3M Medical Department Study: T-63 16.1 3M Environmental Laboratory 3M Environmental Laboratory Page 75 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Protocol LRN-U2103 Amendment Number 1 This amendment modifies the following portion(s) of the protocol: 1. PROTOCORLEADS: The following methods will be used: FACT-M-1.O,Extraction of Perfluorooctanesulfonatefrom Liver for Analysis using HPLC-Electrospray/Mass Spectrometry FACT-M-2.0, Analysis of Liver Extracts for Fluorochemicals using HPLCElectrospray/Mass Spectrometry FACT-M-3.0, Extraction of Perfluorooctanesulfonatefrom Sera for Analysis using HPLC-Electrospray/Mass Spectrometry FACT-M-4.0, Analysis of Sera Extracts for Fluorochemicals using HPLCElectrospray/Mass Spectrometry AMENDTO READ: The following methods will be used: ETS-8-4.1, Extraction of Potassium Perfluorooctanesulfonateor Other Fluorochemical compounds from Serum for Analysis Using HPLCElectrospray/Mass Spectrometry ETS-8-5.1, Analysis of Potassium Perfluorooctanesulfonateor Other Fluorochemicals in Serum Extracts Using HPLC-Electrospray/Mass Spectrometry ETS-8-6.0, Extraction of Potassium Perfluorooctanesulfonate or other Fluorochemical Compounds from Liver for Analysis using HPLC-ElectrospraylMass Spectrometry ETS-8-7.0, Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemicals in Liver Extracts Using HPLC-Electrospray/Mass Spectrometry REASON: The methods originally listed were superseded during the course of the study. 2. PROTOCORLEADS: There is no independent section of the protocol that addresses sample retention. AMENDTO READ: Specimens will be maintained in the 3M Environmental Laboratory specimen archives. Any specimens sent to sub-contract laboratories will be returned to the 3M Environmental Laboratory upon completion of analysis and submission of the subcontract laboratory(s) final report. Specimens analyzed at sub-contract laboratories will be returned with the following documentation: the signed original chain of custody and records of storage conditions while at the sub-contract facility. REAsON: To define in detail the appropriate disposition of specimens analyzed at subcontract laboratories. 3M Environmental Laboratory 3M Environmental Laboratory Page 76 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Protocol LRN-U2103 Amendment Number I 3. PROTOCORLEADS: Section 7 states that the following raw data and records will be retained in the study folder in the archives according to AMDT-S-8: Approved protocol and amendments; approved methods; data summaries; study correspondence; shipping records; raw data; and electronic copies of data. Additionally, Section 7 states that supporting records to be retained separately from the study folder in the archives according to AMDT-S-8 will include at least the following: Approved validation reports; training records; calibration records; instrument maintenance logs; Standard Operating Procedures, Equipment Procedures, and Methods; and appropriate specimens. AMENDTO READ: Section 7 states: "The original data, or copies thereof, will be available at the 3M Environmental Laboratory to facilitate audits of the study during its progress and before acceptance of the final report. When the final report is completed, all original paper data, including: approved protocol and amendments, study correspondence, shipping records, raw data, approved final report, and electronic copies of data will be retained in the archives of the 3M Environmental Laboratory. All corresponding training records, calibration records, instrument maintenance logs, standard operating procedures, equipment procedures, and methods will be retained in the archives of the facility performing each analysis." REASON: To direct subcontract laboratories in the disposition of the items listed above. 4. PROTOCOL READS: The study director for the present study was identified in the protocol as Kristen J. Hansen, Ph.D. AMENDTO READ: The role of study director for the present study was reassigned to John L. Butenhoff, Ph.D., as of 20 January 2000. The previous study director, Kristen Hansen, has been reassigned to the role of Principle Analytical Investigator. REASON: The role of study director was reassigned in an effort to ensure compliance with Good Laboratory Practice Standards that outline study personnel requirements (refer to 21 CFR Part 58). 5. PROTOCOL READS: The sponsor for the present study was identified as Andrew M. Seacat, Ph.D. AMENDTO READ: The role of sponsor for the present study was reassigned to Marvin T. Case, D.V.M., Ph.D., as of 20 January 2000. REASON: The change was made at the request of the sponsor. 3M Environmental Laboratory 3M Environmental Laboratory Page 77 3M Medical Department Study: T6316.1 Amendment Approval Analytical Report: FACT-TOX-001 LRN-U2103 Protocol LRN-U2I03 Amendment Number I Andrew M Seacat, Ph.D., Outgoing Sponsor Representative k l L 6 Kristen J. Hansen, Ph.D., Outgoing Study Director 5!I/Lb/zoaa date / / - F C b -POO Date / b hLL 5L-04 Marvin T.Case, D. K M , Ph.D., Incoming Sponsor Representative Date John L. ButenhoJfjCPh.D., Incoming Study Director F& 2 w y $ l o / Date 3M Environmental Laboratory 3M Environmental Laboratory Page 78 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Study Title 104-Week Dietary Carcinogenicity Study with Narrow Range N-Ethyl PerfluorooctanesulfonamidoEthanol in Rats PROTOCOL AMENDMENT NO. 2 Amendment Date: January 8,2001 Performing Laboratory 3M Environmental Technology & Safety Services 3M Environmental Laboratory 935 Bush Avenue St. Paul, MN 55106 Laboratory Project Identification ET&SS LRN-U2103 FACT TOX-001 Covance 6329-212 3M Medical Department Study:T-6316.1 3M Environmental Laboratory 3M Environmental Laboratory Page 79 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Protocol FACT TOX-001 Amendment No. 2 This amendment modifies the following portion(s) of the protocol: 1. PROTOCOL READS: 1.2 Data collected in the Environmental Laboratory will be considered non-quantitative screening data until future studies have been conducted to determine absolute recoveries of specific or general fluorochemical compounds. AMENDTO READ: 1.2 If matrix spike studies provide accurate representation of recovery of endogenous levels of PFOS, PFOSA, PFOSAA, PFOSEA,M556,and EtFOSE-OH, the 3M EnvironmentalLaboratory will provide semi-quantitative data for sera and liver samples collected from test animals. REASON: Due to improved analytical methods, the decision was made to change the purpose of the study results from non-quantitative to semi-quantitative. 3M Environmental Laboratory 3M Environmental Laboratory Page 80 3M Medical Department Study: T6316.1 Amendment Approval # Analytical Report: FACT-TOX-001 LRN-U2103 Protocol FACT TOX-001 Amendment No. 2 Marvin T. Case, D.V.M., Ph.D., Sponsor Representative /d 9-l */ Date c John L. Butenhog Ph.D., Study Direcior pH JWzaar Date 3M Environmental Laboratory 3M Environmental Laboratory Page 81 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Study Title 104-Week Dietary Carcinogenicity Study with Narrow Range N-Ethyl Perfluorooctanesulfonamido Ethanol in Rats PROTOCOL AMENDMENT NO. 3 Amendment Date: February 15,2001 Performing Laboratory 3M Environmental Technology & Safety Services 3M Environmental Laboratory 935 Bush Avenue St. Paul, MN 55 106 Laboratory Project Identification ET&SS LRN-U2 103 FACT TOX-001 Covance 6329-212 3M Medical Department Study: T-6316.1 3M Environmental Laboratory 3M Environmental Laboratory Page 82 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Protocol FACT TOX-001 Amendment No. 3 This amendment modifies the following portion@) of the protocol: 1. PROTOCOL READS: Specimens will be maintained in the Environmental Laboratory specimen archives. Any specimen sent to sub-contract laboratories will be returned to the 3M Environmental Laboratory upon completion of analysis and submission of the sub-contracted laboratory(s) final report. Specimens analyzed at sub-contracted laboratories will be returned with the following documentation: the signed original chain of custody and records of storage conditions while at the sub-contracted facility. AMENDTO READ: Specimens will be maintained in the Environmental Laboratory specimen archives. Specimens sent to sub-contract laboratories for analysis of PFOS will be returned to the 3M Environmental Laboratory upon completion of analysis and submission of the sub-contracted laboratory(s) final report. Specimens sent to sub-contract laboratories for analysis of compounds other than PFOS will not be returned. Specimens returned by sub-contracted laboratories will be returned with the following documentation: the signed original chain of custody and records of storage conditions while at the sub-contracted facility. REASON: Some specimens sent to sub-contract laboratories will not be returned. Portions of these specimens have been retained at the 3M Environmental Laboratory. 3M Environmental Laboratory 3M Environmental Laboratory Page 83 3M Medical Department Study: T6316.1 Amendment Approval Analytical Report: FACT-TOX-001 LRN-U2103 Protocol FACT TOX-001 Amendment No. 3 Marvin T. Case, D.V.M., Ph.D., Sponsor Representative Date John L. Butenhog, Ph.D., Study Director Date 3M Environmental Laboratory 3M Environmental Laboratory Page 84 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Study Title 104-Week Dietary CarcinogenicityStudy with Narrow Range N-Ethyl Perfluorooctanesulfonamido Ethanol in Rats PROTOCOL AMENDMENT NO. 4 Amendment Date: April 2,2001 Performing Laboratory 3M Environmental Technology & Safety Services 3M Environmental Laboratory 935 Bush Avenue St. Paul, MN 55106 Laboratory Project Identification ET&SS FACT TOX-001 3M Laboratory Request No. U2103 3M Environmental Laboratory 3M Environmental Laboratory Page 85 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Protocol FACT TOX-001 Amendment #4 This amendment modifies the following portion(s) of the protocol: 1. THEAMENDED PROTOCOL READS: (Amendment #1) The following methods will be used: ETS-8-4.I , "Extraction of Potassium Perfluorooctanesulfonate or Other Fluorochemical Compounds from Serum for Analysis Using HPLC-ElectrosprayMass Spectrometry" ETS-8-5.I , "Analysis of Potassium Peffluorooctanesulfonateor Other Fluorochemicals in Serum Extracts Using HPLC-ElectrosprayMassSpectrometry" ETS-8-6.0, "Extraction of Potassium Perfluorooctanesulfonate or Other Fluorochemical Compounds from Liver for Analysis Using HPLC-ElectrosprayMass Spectrometry" ETS-8-7.0,"Analysis of Potassium Perfluorooctanesulfonateor Other Fluorochemicals in Liver Extracts Using HPLC-ElectrosprayMassSpectrometry" AMENDTO READ: The following methods will be used: ETS-8-4.1, "Extraction of Potassium Perfluorooctanesulfonate or Other Fluorochemical Compounds from Serum for Analysis Using HPLC-ElectrosprayMass Spectrometry" ETS-8-5.I , "Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemicals in Serum Extracts Using HPLC-ElectrosprayMass Spectrometry"with thefollowing exception: (Section 14.5.1) "Matrix spike recoveries must be within -F 30% of the spiked concentration for all analytes except M556, which must be within 6 0 % . ETS-8-6.0, "Extraction of Potassium Perfluorooctanesulfonate or Other Huorochemical Compounds from Liver for Analysis Using HPLC-ElectrosprayMas Spectrometry" ETS-8-7.0, "Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemicals in Liver Extracts Using HPLC-ElectrosprayMass Spectrometry" with the following exception: (Section 14.5.1) "Matrix spike recoveries must be within +30% of the spiked concentration for all analytes except PFOS, which must be G O % . REASON: The analytical method and resulting QC data support a 50%-150% acceptable range for spike recoveries for these analytes and matrices. 3M EnvironmentalLaboratory 3M Environmental Laboratory Page 86 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Protocol FACT TOX-001 Amendment #4 Amendment Approval John L. Butenhoff, Ph.D., Study Director Date Marvin T. Case, D.KM., Ph.D., Sponsor Representative Date 3M Environmental Laboratory 3M Environmental Laboratory Page 87 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Record of Deviation 1 Study / Project No. 1. FACT-TOX-001 Identification Covance 6329-212 Deviation Type (Check one) 0 SOP X Method 0 Equipment Procedure 0 Protocol 0 Other: _ _ _ _ I - ____ ___ Document N&ber(s): Date(s) of occurrence: FACT-M-2.0, FACT-M-4.0, ETS-8-5.0, Entire Study ETS-8-7.0 11. Description: Required Procedure/process: These methods state: Typically the analytical batch run sequence begins and ends with a set of extracted matrix standards. Actual Procedure/process: In ma-ny analytical r- uns, only the initial extra-c--ts-_matrix curve - was use-d to evaluate the extracts. "_I_ I __I__ __l_l_--_ Either the second curve was not injected or the bracketing curve was deactiv.a"te- d.s-.inc_e_th- is calibraton curve was divergent. I Ill. Actions Taken: (such as amendment issued, SOP revision, etc.) This deviation was written and a few method modificationswere also written stating that the second bracketing calibration curve may be deactivated if ins&ental drift affects the data. The first curve and acceutable calibration checks shall then bracket usable data. H Recorded Bv Date ~ ~ & 5 //-5---- p IV. Impact on Study / Project - .- - No adverse impact on the outcome of this study shce data weresufficiently_ b_ r-a-cketed by ca_l_i-bra_t_io- n c-h_ec-k-s.-- __ ----_ - - -- - -l_l-_--"_I I _ _ ___ _ I _- - - Authorized By (Study Director /Project Lead) Date 4J L . 2 - &2&&4gf&// 3M Environmental Laboratory FOKWETS-4-8.0 3M Environmental Laboratory Deviation No. 1 (assigned by Study Director or Project Lead at the end of study or project) Page 88 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Record of Deviation 1. Identification ______ _ I- " ~ Study I Project No. FACT-TOX-001 Covance 6329-212 --__-_____---__ Deviation Type (Check one) a SOP DProtocol x Method IJEquipment Procedure iJ Other: - ~ __ Document Number(s): FACT-M-3.0 Date(s) of occurrence: 04123198,04128198,05111198 11. Description: R......equired F'rocedure/process: ................... .......... ........... ____ ._ ............ ................ asthe.s.amp1es, S...e....c....t..i...o....n.......1....1......1.2 s..t..a..t..e...s..: If the major..i...t..y of th.....e serum sample-s.__a_r_e-.._less than 1.O mL, ............................................... extra...c....t the standards ........................... in the_S~.e.,v.o~u,m.e.. ..........____ ~~ __ .. __.__c Actual Procedure/process: _-___~__"-l___ .I..n...i.t..i.a...l...s..e...r..a...v.. olume.s for weeks ...... 4, 8, and 14.....w_er_ e below 1..O mL (0.6 0.5 mL, mL,........................... & 0.5 m_L_re.spectiv.e..l.y...). T. -h.e_c_u_r~vIe_s....prepared fo__r_t._h_e_-s_e samples were made using 1.O mL of serum. .___I____ I_ _ __ _ _--. ................. .................... ..___ __ ......................................................... ~~ ...................... Ill. Actions Taken: (such as amendment issued, SOP.rev- ision, etc.)_.__-_ ___ _ _____-. This-deviationwas G-tten. -- -- -- _ - - . -_ __ 3M Environmental Laboratory Form ETS-4-8.0 3M Environmental Laboratory Deviation No. a (assigned by Study Directoror ProjectLead at the end of study or project) Page 89 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Record of Deviation 1. Identification - - - - ~ - ~ I _ - _ _ S m P s c x F A C T - T O X - 0 01 Covance 6329-212 Deviation Type (Check one) a SOP DProtocol x Method Cl Equipment Procedure a Other: Document Number(s): ETS-8-5.1 . Date(s) of occurrence: 05/05/98 I/* Description: Required Procedure/process: ........ -_____ ___._ __ . ....................................... _. Actual Procedure/process: - _" _____-I_______ ~ ____I 4/2-8-/-98) analyzed on 05/05/9-8--wa-s ab-o_ve_th_e_ra_n_ge-of-th-e calibration curve. .... ........................ . - - ......... ......... ............... .- ........... ....... .- ............... ... .-....... .................... - ............................ -.... 111. Actions Taken: (such as amendment issued, SOP revision, etc.) I__ -- ~- ___ This deviation was yitten.- - - -~ __ IV. Impact on Study / Project .Th~e s_ am_pleIre_sult will be I - _ll.___l_ll _ _ I " ~ ~~~ "^l -- -- Authorized By (Study Director /Project Lead) .I I Date i .- ~ .-. - 3M Environmental Laboratory Form ETS-4-8.0 3M Environmental Laboratory Deviation No. 3 (assignedby Study Director or Project Lead at the end of study or project) Page 90 3M Medical Department Study: T6316.1 Record of Deviation Analytical Report: FACT-TOX-001 LRN-U2103 1. Identification Study / Project No. a SOP X Method TOXOO1 (LIMS #U2103) ~ l__ll____ Deviation type ~ _ _ _ I I _ _ _.____ (Check one) ___~_ -- 0 Equipment Procedure - _- --- OProtocol Cl Other: _ I - - I _ - - _ _ Document number FACT-M-4.0 and ETS-8-5.1 Date(s) of occurrence I 05/05/98,4/20/00,4/21/00, 5/02/00, 5/24/00, j 5/30/00, 8/23/00,02/28/00, 03/15/00, i 02/27/00,04/17/00,04/18/00,04/19/00, 05/31/00,06/19/00, 06/20/00,06/2 1/00, 08/02/00,04/06/00 11. Description Required procedure/process: Section 10.1.2: Analyze a method blank and a matrix blank prior to each calibration curve. ~ 1 1 Actual I procedure/process: ~- l _ - - _ _ l - -On- se-ve-ral~occasions, the blanks were either a) analyzed after the calibration curves orb) not analyzed at all for a particular MS run. ~_-~_ _lll_--_-_l____ll_l---_ ~ _ - I 111. Actions Taken (such as amendment issued, SOP revision, etc.) Deviation written. -I_~_l_l___-I_ Recorded by ~ l _ l _ _ _ IV. Impact on Study / Project (completedby StudyDirector or Project Lead) ............................ - __ ~ ...... All blanks were analyzed at least once during the course of the study. When blanks were run after the calibration curve, all control-of-bias practices were followed. In the instances where blanks were not run at all, the samples being analyzed were usually well above the typical run LOQ. In addition, the vast majority of samples run without blanks were high level dilutions. The hnction of the blank is to determine if contaminationwas introduced during extraction; any low-level contamination would be inconsequential to extracts requixjng dilution. This deviation has no adverse affect on the study data. ............................................... ........... ............. .- ..... ............ .- ........ - ............................................................ ................ - ... 4h Oe/l..O../O.I. ~ ._....... .. ........ ... ................... . _ .. __ - ..... ._ _ ............._ Authorized by -. .- 1 - --- --___ __ 3M Environmental Laboratory Page 91 3M Medical Department Study: T6316.1 Record of Deviation Analytical Report: FACT-TOX-001 LRN-U2103 1. Identification Study / Project No. a X T..-O-.""Xl__O_"O_.l__.(_LI_-I.-MS #U2103) Covance 6329-212 "_ .-.---" --.-I_. ~~ ~ Deviation type SOP Method (Check one) ~ ~ 0 Equipment Procedure -- _i - - - I - - - - - CJProtoc- ol CI Other: I I _ ~~- ~ ~- l l - _ l _ - l - _ Document number: ETS-8-5.1 (Modified method) Date(s) of occurrence: ETS-8-7.0 (Modified method) 05/05/98 (ETS-8-5.l), 02/18/99, 12/06/00 (ETS-8-7.0) 11. Description Required procedure/process: -_ Section 14.3.6: A valid calibration curve must contain at least 5 active Doints ~- Actual procedure/process: -_-- l_ll_l~-_l_-l--l A suitable calibration curve (i.e. with all points within 30% of theoretical and with an r"2>0.980) could not be derived from 5 points; four points were used for the calibration curve for PFOS and PFOSAA (5/5/98), for Et-FOSE-OH (2/18/99), and for PFOSAA (12/06/00). Ill. Actions Taken (such as amendment issued, SOP revision, etc.) - - _ -- ~ II I_----Ix ll_- I I_--I1-__ ___ __--_^-_- _I_--_I Deviation written. Data evaluated against 4 point curve is specificallymarked in the table of final results. IV. Impact on Study f Project (completed by Study Director or Project Lead) ____ - __ The data quality __ .of_s_amples evaluated against - - -- the f- o_z _p o i n t curve i-s ad__v-e_r_s_e_ ly affe_c_ted. Authorized by -- - Date 3M EnvironAmtteanchtamleLntaAb:oRraectoorrdyof Deviation ETS-4-8.0 Page 92 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Record of Deviation 1. Identification - Study / Project No. FACT-TOX-001 Cov&ce 6329-212 -- . __ IDeviation Type (Check one) - - __ Document Number(s): FACT-TOX-001 0 SOP 0 Method 0 Equipment Procedure XProtocol 0 Other: .- -- - Date(s) of occurrencei 05/06/98,05/13/98,05/14/98,05/15/98,. 05117/98,06/03/98, 06/05/98,06/12/98, 06/14/98,06/18/98,06/25/98,06/26/98, 07/06/98,07/09/98, 12/14/98, 12/29/98, 01/06/99,0 1/18/99,02/16/99,02/18/99 11. Description: Required Procedurelprocess: IThe protocol states: Methods FACT-M-2.0 and FACT-M-4.0 are to beused to analyze liver and sera extracts. -_ - ~ P Actual Procedure/process: Methods were not documented or the incorrect methodswere listed for-analyses of these I Ill, Actions Taken: (such as amendmeIn-t_issu-ed,-_SO_P_ revisio_n_,_etc.) - - - -- - _I_I_ I This deGation was written. - I ,.. ,.. -. " - RecordedBy , . . , , . ,I_ _ _ - . ~-.-....-...~,"."__I___" __^.--.._I_ I_____._ _ I I ~ .... . . . li "I . "~ . . , : Date dYA. L IV. Impact on Study / Project T.hese an-alyses were review-- ed an- d-th-_e.s_e-_da_ta were ana@ed accordingto - m-e- thods as -li-sted in -_I --__I-__ the protocol. No adverse impact on the outcome of this study. _Authorized By (Study Director /Project Lead) -~ - ~ Date s%, &hr DufChhoff 3M Environmental Laboratory Form ETS-4-8.0 3M Environmental Laboratory 1 , y ~ h s s v:M a u c h GYL3@t9w Deviation No. (assigned by Study Director or Project Lead at the end of study or project) Page 93 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Record of Deviation I 1. Identification Study / Project No. -FACT'-TOX-001 Covance 6329-212 Deviation Type (Check one) __ - __ 0 SOP x Method- __ c1 Equipment Procedure OProtocol lJ Other: __ Document Number(s)r ETS-8-5.1 _- - Date(s)-of occurrence: I 05/14/98 11. Description: .Require-d. Procedure/process: .- __.-_-._____ ................................................. ___ . ............................................ The continuing calibration verification (CCV) must be within +/- 30~ % o~ f thx e t_ heo- ret- ica- l _ .l_ ll_. - - concentration. .... ..................... ....... ..._ . ...... __ . .......:....... . ......... ...................................... IActual Procedure/process: ..... ....................................................................... .. -.. . ........ ........... __ ......... PFOSAA The Week 8, Sera, PFOS and .... .. MS, ....... MSD, H 2 0 Blank, an.d..... S_ _e.r....a.... Blank . data .. were ................. entered even ............................................. though ....... CCVs ... d_i_d_n....o.....t.....m........e.....e....t......c....r....i..t...e. ria. _. .......... ..................... ............................................. The CCV analyzed prior to the PFOS data was within criteria and the curve immediately ........... ...... . ...... . .-.__ .............................................. ........ .. ................... -. ...................................... following these data were within criteria. Three CCVs in a row, prior to the good CCV, failed .-._I" " .. I-----. .I -" lllll___l._----.-- ... --__1__1__ the 30% criteria. _-- __ -- - Two CCVs analyzed prior to the PFOSAA datawere within criieria and the Grve ........ .................... . .. ........... ................................................. . -..... .. . . . . .............. .......... ..........._.. . immediately foliowing these data were within criteria. ........... . .... ............... ...- .... Two CCVs prior to the .......... ......... .- go_....o.. d CCVs ......... ........ ~ failed the 30% criteria. I....- ..... ___ ....... . . .... _ ... ............ . Ill. Actions Taken: -. ...... ................... (such as a- me.n....dment issued, SOP r_ev_isi-o. n, etc.) ~ - ~ ... .... ..................................... This deviation was written and data will be flagged as tentative in the raw data. ...... Recorded By &&A ___ .................................... cQcrrL,L, IV. Imroact on Studw / Proiect : Date................. .............................. 3M Environmental Laboratory Form ETS-4-8.0 3M Environmental Laboratory Deviation No. 3 (assigned by Study Director or Project Lead at the end of study or project) Page 94 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Record of Deviation 1. Identification Study / Project No. FACT-TOX-001 Covance 6329-212 Deviation Type (Check one) 17 SOP 13Protocol x Method tJ Equipment Procedure Cl Other: Document Number(s): ETS-8-7.0 Date(s) of occurrence: 05/29/98,06/01/98 11. Description: Required Procedurdprocess: .......................... ........................................................... . -. A... five..p..o..i.nt cu.r.ve or the ca..li.b--ra..t.i..o..n. curve. ._._......... ... .................. ...................................................................................... .............................. . ........ ........ .... Actual Procedure/process: -- A fourpoint curvewas used for plo&ng the EtFOSE calibrationcurvefor the Weekl4, Liver curve, generated on 12/14/98. ............ ................. .... ............. Ill. Actions Taken: (such as amendment issued, SOP revision, etc.) These data were flagged in the raw data and this deviation was written. .................................... ......................... ................... ............. -.. Recorded By ............. Date EtFOSE:OHda&s IV. Impact on Study/ Project - flagged as qual%itive, so thisdata &ill not be adversely affected. hh o ~ / / ~ / O / Authorized By- (Study Director /Project G a d ) - .Date 3M Environmental Laboratory F o ET~S-4-8.0 3M Environmental Laboratory Deviation NO. 8 (assignedby Study Director or ProwLead at the end of study or project) Page 95 3M Medical Department Study: T6316.1 ~~ ~~ Analytical Report: FACT-TOX-001 LRN-U2103 Record of Deviation I. Identification Study / Project No. FACT-TOX-001 Covance 6329-212 Deviation Type (Check one) D SOP 13 Protocol x Method D Equipment Procedure D Other: Document Number(s): ETS-8-7.0 Date(s) of occurrence: 06/02/98,01/14/99,01/17/99 11. Description: Required Procedure/process: - . Section 14.4.1 states: matrix spikeDercent recoveriesmustbe within-&30%ofthe suiked concentration. _ _ __ -- .- Actual Procedure/process: The week 14,liver matrix spike &d matrix spike duplicate samples prepared for PFOS apd PFOSAA analysis(exQacted 06/02/98) weren't within criteria. -me noncompliance-of the-PFOS and PFOSAA data on 06/02/98 can-be agributed to inappropriatepreparation: spikes were prepared at lOOng/g, a level not distinguishablefrom background levelspf the analytein the-controls. Spikes were reprepped at a higher level (250 ndg) on 01/14/99, but again, for PFOS analysis, the spikelevel was not distinguishable from background levels; PFOSAA was inadvertently omitted from analysis. Spikesfor the week 1_4liver sampleswere prepared a third time on-O1/17/99 at 750 ngg. Results-ofthe third analysis show@ an averagePFOS recovery of 132%;one of the two matrix spikes was recovered outside the stated 130%criteria (138%). Results of matrix spike recoveryfrom PFOSAA at the 750 ng/g level were acceptable. Ill. Actions Taken: (such as amendment issued, SOP revision, erc.) This deviationwas &itten. A note will be added tothe week14 liver data stating that one PFOS MS was recovered at 132% and that week 14 PFOS analytical results may be biased high. Recorded By -_ . __ - Date IV. lmpaa on Study/ Project PFOS results for week 14liver samples may be biased high. Authorized By (Study Director /Project Lead) $jb O Z / l 4 / 6 / Date 3M Environmental Laboratory F o ET~S-4-8.0 3M Environmental Laboratory Deviation NO. 9 (assigned by Study Director or Project Lead at the end of study or project) Page 96 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Record of Deviation 1. Identification Study / Project No. FACT-TOX-001 Covance 6329-212 Deviation Type (Check one) a SOP CJ Protocol x Method 0 Equipment Procedure Cl Other: Document Number(s): ETS-8-7.0 Date@)of occurrence: 06/03/98 11. Description: .R...e..q..u..i..r.e..d....P..rocedur.e../..p..r..o..c..e. ss: ....... ................ ... ... ..... .S..e..c..t.i.o...n...1..4....4....1...states:. . .m. . a. .t.r.i.x. . .s.p. .i.k. .e. .p. .e.r. c. .e. .n. t. .r. e. .c. .o.v. .e. .r.i.e. .s. .m. .u. .s.t. .b. .e. . w. . .i.t.h. .i.n. .&. . 30% . . . . . . . .o. .f. .t.h. .e. .s. .p. i. k. .e. .d. . . . . . . .c-o.n..c.e-n.t.r.a. t-io..n... -. ....- ...-.-. .... - ...-.- .... .......... .... - .. .- .... - .. - .- ....- _ .... _...._ ... _ _.... - ...... Actual Procedure/process: - _- All week 4 liver MS recoveries are outside criteria, however, themat- rix . spike-and s&pie d a b appeared to be mislabeled. When the MS and sample labels are corrected, all recoveries are within critega-withthe Exception of PFOS and EtFOSE. Average PFOS recoveries are 192% and average EtFOSE Recoveries are 133%. -. . . . . . . . . . . - . ................ _. . .... ..... ....... ~ Ill. Actions Taken: (suchas amendment issued, SOP revision, etc.) The MS and sample data labels were corrected in the raw data, the PFOS data were flagged in the spreadsheet, sample results are noted to have a potential high bias, &d 6 s deviation was written. - _.- Recorded By Date IV, Impact on Study / Project PFOS data in week 4 liver samples is noted to be potentially biased high. EtFOSE-OH data 3;i Are already d e s i p a t e as qualitative only and is not fu-&er-affected by the deviantrecoveries. o=./M/01 .-. Authorized By (Study Director /Project Gad) - Date 3M Environmental Laboratory F o ET~S-4-8.0 3M Environmental Laboratory Deviation No. lo (assigned by Study Director or Project Lead at the end of study or project) Page 97 3M Medical Department Study: T6316.1 Record of Deviation Analytical Report: FACT-TOX-001 LRN-U2103 Study / Project No. FACT-TOX-001 - __ Covance 6329-212 IDocument Number(s): a Protocol ETS-8-7.0 II. X Method o Other: Equipment Procedure Date(s) of occurrence: 12/29/98 Description: Required Procedurelprocess: Section 14.5.1states:Continuing calibrationvenficationpercknt recoveries mustbe within +/-30% of the soiked concentrationl - _- __ __ __ I .. - . .. - - . . . __ - ._.. -~.... . . . -. . - . . _. Actual Procedure/process: The continuingcalibrationverification s6plesfor PFOS; PFOSA,A d PFOSAA analyzed on 12129-98 .had percent recove-es greater- ~ a1q30.%of-expected. ___ ___ . _- - - _ _ - - _- I __ - .- __ - - ._ __ __ - - -- _- __ - - - __ __ __ -. __ -_ Ill. Actions Taken: (such as amendment issued, SOP revision, etc,) This deviation was written; affected data is noted in the data tables - -. . . - .~ ~. ... . .-. . __ . . .- . ..~.. Recorded By n IV. Impact on Study / Project Affected data may be biased high. - _c,'b. bz((3IbI -~ .... . -. . ._.. . ... . - Date 2. '3-0( -I_._ 3M Environmental Laboratory Form ETS-4-8.0 3M Environmental Laboratory Deviation No. 11 (assigned by Study Director or Project Lead at the end of study or project) Page 98 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Record of Deviation 1. Identification Study / Project No. FACT-TOX-001 Covance 6329-212 Deviation Type (Check one) a SOP Protocol x Method b Equipment Procedure 0 Other: Document Number(s): ETS-8-7.0 Date(s) of occurrence: 01/18/99 11. Description: Required Procedure/process: -- Section_ 1_ 4.5_ .1 s.ta- tes: Continuing calibration verification percent recoveries must be within ~ Actual Procedurdprocess: The-Week 14, liver samples C91281F, C91288F, d91293F, C91299F, and C91304F were bracketed by a-continuingcalibration verification that did not meet the +/- 30% criteria. (-31%) Ill. Actions Taken: (such as amendment issued, SOP revision, etc.) These data were flagged in the raw data as having a potential low bias and this deviation was written. Recorded By dih b LIV. lmpact on -Study/ Project Resuits for these analyses may be-biased low. - . . .. .. .. .. . .. .. . - .. . .. - . . . . . . ..- .. . .. . . . .-.- ~ . . . .~ . . . . . . ... . Date .. . Authorized By (StudyDirector /Project Lead) Date 3M Environmental Laboratory Fomt ETS-4-8.0 3M Environmental Laboratory I' Deviation No. la (assigned by Study Director or Project Lmd at the end of study or project) Page 99 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Record of Deviation 1. Identification Study / Project No. FACT-TOX-001 Covance 6329-212 Deviation Type (Check one) Docum&t-Nu&&( s ) I FACT-TOX-001 0 SOP 0 Method 0Equipment Procedure XProtocol 0 Other: - -_ -- __ - Date@)of occurrence: 02/07/00,02/08/00,02/09/00 11. Description: Required Procedure/process: ... .ll._l I ....._I_____" _. _ I . ,.. _ _ _ I . ........ ~.......~~..._._....I" ." ......-..."I . The lxotocol states: Method FACT-M-3.0 is to be used when extracting the serum samples. -. ... Actual Procedure/process: Method ETS-8-4.1 was used to extract these serum samples. - I " ".I . . . .. . .. ~ , . ". ...... "_ Ill. Actions Taken: (such as amendment issued, SOP revision, etc.) This deviation w& written,'- Recorded By __ - .__ IV. Impact on Study / Project 4 No adverse impact on the outcome of this study. Sera method ETS-8-4.1 is an improvement over sera meth-o_ _d FAC_ T_ -M- --3~.O.. ;S b 5/W/Q) Authorized By (Study Director /Project Lead) Date 5$+ 1 3M Environmental Laboratory Form ETS-4-8.0 3M Environmental Laboratory Deviation No. 13 (assigned by Study Director or Project Lead at the end of study or project) . Page 100 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Record of Deviation 1. Identification Study / Project No. FACT-TOX-001 Covance 6329-212 LIMS U2103 Deviation Type (Check one) 0SOP 0Protocol x Method 0Equipment Procedure 0 Other: Document Number(s): ETS-8-7.0 Date(s) of occurrence: 02/22/00,04/24/00, 04/26/00,04/28/00, 05/24/00,07/28/00, 08/01/00,08/14/00,08/15/00, 08/21/00, 08/23/00,03/22/01 11. Description: Required Procedure/process: _-- _I^____II--- - ^_x____ - -I___----I.- - ___I I _ .I_--_^__ _ _ _ __I__^_I__ Section 10.1.2 states: Analyze a method blank and a matrix blank prior to each calibration curve. - - . .. .. -. .- . ..- . ....... . - . . .. .. .- - -. - .... . I"..- .. . ... .. . . . .- . - ,. . _ _ ,, Actual Procedure/process: On several occasions,the bla& were eithe.r a).ana-ly.z.ed.af.t.er the calibration curves or b) n-o.t analy.ze.dAt all for a particular MSanalysis. 111. Actions Taken: (such as amendment issued, SOP revision, etc.) This deviation was written. 1 RecordedBy Date IV. Impact on Study/ Project 1 All blanks were analyzed at least once during the c o k e of the study. When blanks were analyzed . . after the calibrationcurve. allcontrol-of-biasDracticeswere followed. In the instanceswhere blanks - Iwere not analvzed at all. the samdes being aialvzed were usuallv well above the mica1run LOO. Y -.- 4 In addition, tce vast majoritfof gamples analyzed without blank; were high 1 function of the blank is to determine if contaminationwas introduced during extraction; any low-level contamination would be inconsequential to extracts requiring dilutions. This deviation has no adverse affect on these study data. - 4 h OY//4lO/ IAuthorized By (Study Director /Project Lead) Date 3M EnvironmentalLaboratoiy Form ETS-4-8.0 3M Environmental Laboratory Deviation No. 1y ' (assigned by Study Director or Project Lead at the end of study or project) Page 101 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Record of Deviation " 1. Identification - .............. ......... ..... . Study / Project No. : ala -t4..k-t-SoX-..oo.!.. C.O~U.G?rcQ 6..2a.9-- . . .... ..... ..... a a Deviation Type SOP El Method Eq-ui-pment Procedure a (Check one) - Protocol t i Other: .. ........ Document Number ETS- 8 -5. I ............ .- ................................ 11. .... ~ ._ i Date(s) of occurrence i i OLl2llDO Description: Required Procedure/process: d ....Ganm+rQ..kbn .......... I ....... I . __..___ Actual Procedure/txocess: _ I _ Recorded By I -_____.______ Date U6i2lo4a IK Impact on Study/ Project - _ I _ I - Authorized By (Stud' D F o ET~S-4-8.0 3M Environmental Laboratory ---___I-.---- w S p t W . R-p: b d CaLC : Date (assigned by Study Director or Project Lead at the end of study or project) Page 102 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Record of Deviation 1. Identification "" Study / Project No. FACT-TOX-001 Covance 6329-212 Deviation Type (Check one) - 0 SOP [IMethod [IEquipment Procedure XProtocol 0 Other: FACT-TOX-001 07/28/00,08/01/00,08/14/00,08/15/00, 08/2 1/00,08/23/00,08/28/00,08/29/00 11. Description: IRequired Procedure/process: Theprotoco_l _states: Metho_d- -ET-S-8-7.0 is to be used to analyze liv--er extr_%_cts. IActual Procedure/process_: _ Method ETS-8-6.0was incorrectly listed as the method used to analyze these liver extracts. ' I .... . -.......~ ...................... ................................................ ..... .............. .............. Ill. Actions Taken: -(such as amen--dment issu-ed-, SO-P rev~bio-n--,_etc- .) -_1__--- _I - I _ __ I_ This deviation was written. - - - I_ I RecordedBy -. Date . IV. Impact on Study / Project ET-S_~-8-6-.0-__is__t_h_e--extractionmethod, ETS-8-7.0 is the analytical method-hese analyses--were -__--I_-_I ~ I _ - _ _ _ I ~ -____^I_- _ _ -him y o f reviewed and data were analyzed acc_o-rd-i-n-_g to ETS-8-7.0. No adverse impact o-n the outcome _ - I - _ _ _ I _I____---- this of study. _- - I I___I__ - - I_ _- I I I -- _-_ I Authorized By (Study Director /Project Lead) -. Date I' S p :&hn BMfLllhofl 3M Environmental Laboratory Form ETS-4-8.0 3M Environmental Laboratory (assigned by Study Director or Project Lead at the end of study or project) Page 103 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Record of Deviation 1. Identification Study / Project NO. .-- . .. Deviation Type (Check one) __ #SOP OProtocol PMethod Cl Equipment Procedure iJ Other: -.. -- -- Document Number p- -7 0 I Date(.s,) of occurrence i ,3-3!?2&3/ II. Description: m !.-mJ&A Req...u.._ir_ed Procedure/process: . ..... . ........... lL.L%4uq ...... .............................. .A... ctual Procedure/process: ...... .............. ............ ...................................................................................... ...... - _ - ~ _ ^ ................. ....... _I^ Ill. Actions Taken: (such as amendment issued, SOP revision, etc.) ................................. ................................. __.-._^--I ..... Authorized By (StudyDirector 1Project Lead) ................ -7- ..h 4.3L2 \ Date .- _- .-...... 3M Environmental Laboratory F o ET~S-4-8.0 Sbd8 ~ ; ~ ) o -,.-. &ob, 3M Environsm~~e~n~t~a~lRLeapb-\ocrhae:tory &AJC Mtxivin Cade k-rL Deviation NO. 17 Director or Project Lead at the end of study or project) 27btd%, Page 104 3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Analytical Report: FACT TOX-001 LRN-U2103 Appendix C: Extraction and Analytical Methods This appendix includes the following methods: Preparatory Methods FACT-M-1.O. Extraction of Potassium Perfluorooctanesulfonateor Other Anionic Fluorochemical Surfactants from Liver for Analysis Using HPLC-Electrospray/Mass Spectrometry, (8 pages) FACT-M9.0, Extraction of Potassium Perfluorooctanesulfonateor Other Anionic Fluorochemical Surfactants from Serum for Analysis using HPLC-Electrospray/Mass Spectrometry, (8 pages) ETS-8-4.1, Extraction of Potassium Perfluorooctanesulfonate or other Fluorochemical Compounds from Serum for Analysis using HPLC-Electrospray/MassSpectrometry, (14 pages) ETS-8-6.0, Extraction of Potassium Perfluorooctanesulfonateor other Fluorochemical Compounds from Liver for Analysis using HPLC-Electrospray/MassSpectrometry, (14 pages) AnalyticaI Methods FACT-M-2.0, Analysis of Liver Extracts for Fluorochemicals Using HPLCElectrospray/Mass Spectrometry, (8 pages) FACT-M-4.0, Analysis of Fluorochemicals in Serum Extracts Using HPLCElectrospray/Mass Spectrometry, (8 pages) ETS-8-5.1, Analysis of Potassium Perfluorooctanesulfonateor other Fluorochemicals in Serum Extracts Using HPLC-Electrospray/Mass Spectrometry, (9 pages) ETS-8-7.0, Analysis of Potassium Perfluorooctanesulfonateor other Fluorochemicals in Liver Extracts Using HPLC-Electrospray/Mass Spectrometry, (10 pages) 3M Environmental Laboratory Page 105 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 3M ENVIRONMENTLAALBORATORY METHOD EXTRACTIOONF POTASSIUM PERFLUOROOCTANESULBONATEOR OTHER ANIONIC FLUOROCHEMICASULRFACTANTS FROM LIVERFOR ANALYSIS USING H P L C - E L E C T R O S P R A YSP~EC~TSRSOMETRY Method Number: -FACT-M-1.0 1 Author: Lisa Clemen Approved By: 81%- 1 % - Laboratory Manager Group Leader TGhnical Reviewer Adoption Date: 5 / J L / 9 ,f Revision Date: M)A Date 3/2l, / q f Date 5123)4P Date fa l 1.0 SCOPE AND APPLICATION 1.IScope: This method is for the extraction of Potassium Perfluorooctanesulfonate (PFOS) or other fluorochemical surfactants fiom liver. 1.2 Applicable Compounds: Fluorochemical surfactants or other fluorinated compounds. F k i c r o s o f t 7.0.1/95 - 3M Environmental Laboratory FACT-M- 1.O Extraction of PFOS from Liver Page 1 of 8 Page 106 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 2.0 SUMMARY OF METHOD 2.1 This method describes how to extract potassium perfluorooctanesulfonate (PFOS) or other fluorochemical surfactants from liver using ion pairing reagent and 5.0 mLs of ethyl acetate. An ion pairing reagent is added to each sample and partitioned into ethyl acetate. Four mLs of extract is removed to a centrifuge tube and put onto a nitrogen evaporator - until dry. Each extract is reconstituted in 1.O mL methanol then filtered through a 3 cc plastic syringe attached to a 0.2 pm filter into glass autovials. 4.1.1 Use universal precautions when handling animal livers, they may contain pathogens. , 5.0 INTERFERENCES 5.1 There are no known interferences at this time. 6.0 EQUIPMENT 6.1 The following equipment is used while carrying out this method. Equivalent equipment is acceptable. 6.1.1 6.1.2 6.1.3 6.1.4 6.1.5 6.1.6 Ultra-Tmax T25 Grinder for grinding liver samples Vortex mixer, VWR, Vortex Genie 2 Centrifuge, Mistral 1000 or IEC Shaker, Eberbach or VWR Nitrogen Evaporator, Organomation Balance 7.0 SUPPLIES AND MATEIUALS 7.1 Gloves 7.2 Dissecting scalpels 7.3 Eppendorf or disposable pipettes 7.4 Nalgene bottles, capable of holding 250 mL and 1 L 7.5 Glass, type A, volumetric flasks 7.6 40 mL glass I-CHEM vials 7.7 Plastic sampule vials, Wheaton, 6 mL 7.8 Polypropylene centrifuge tubes, 15 mL 7.9 Labels 3M Environmental Laboratory FACT-M-1 .O Extraction of PFOS from Liver Page 2 of 8 Page 107 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 7.10 Syringes, capable of measuring 10 pL to 50 pL 7.11 Glass, type A, volumetric pipettes 7.12 Graduated pipettes 7.13 Electronic pipettor, Eppendorf or equivalent 7.14 Timer 7.15 Disposable plastic 3 cc syringes 7.16 Filters, nylon syringe filters, 0.2 pm, 25 mm 7.17 Crimp cap autovials Note: Prior to using glassware and bottles, rinse 3 times with methanol and 3 times with Milli- Q" water. Rinse syringes a minimum of 9 times with methanol, 3 rinses from 3 separate vials. 8.0 REAGENTS AND S~~ANDARDS 8.1 Reagents 8.1.1 Sodium Hydroxide (J.T Baker or equivalent), (NaOH) 1ON: weigh approximately ' 200 grams NaOH. Pour into a 1000 mL beaker containing 500 liters (L) Milli-Qm water, mix until all solids are dissolved. Store in a 1 L nalgene bottle. 8.1.2 Sodium Hydroxide (J.T Baker or equivalent), (NaOH) 1N. Dilute 10N 1:lO. Measure 10 mL of the 1ON NaOH solution into a 100 mL volumetric flask and dilute to volume using Milli-QTMwater. Store in a 125 mL nalgene bottle. 8.1.3 Tetrabutylammoniumhydrogen sulfate (Kodak or equivalent), (TBA) 0.5M:Weigh approximately 169 grams of TBA into a 1 L volumetric containing 500 L Milli-QTM water. Adjust to pH 10 using approximately 64 mL 1ON NaOH and dilute to volume with Milli-Qm water. Add NaOH slowly while adding the last 1 mL of NaOH because the pH changes abruptly. Store in a 1 L nalgene bottle. 8.1.3.1 TBA requires a check prior to each use to ensure pH = 10. Adjust as needed using 1N NaOH solution. 8.1.4 Sodium carbonate/Sodium Bicarbonate Buffer (J.T. Baker or equivalent), (NqCO,/NaHCO,) 0.25M: Weigh approximately 26.5 g of sodium carbonate (Na,,CO,) and 21 .O g of sodium bicarbonate (NaHCO,) into a 1 L volumetric flask and dilute to volume with Milli-QTMwater. Store in a 1 L nalgene bottle. 8.1.5 PFOS (3M Specialty Chemical Division), molecular weight = 538. 8.1.6 Ethyl Acetate, Omnisolv, glass distilled or HPLC grade. 8.1.7 Methanol, Omnisolv, glass distilled or HPLC grade. 8.1.8 Liver and control liver, received frozen from testing laboratory. - 8.1.9 Milli-QTMwater, all water used in this method should be Milli-QTMwater and may be provided by a Milli-Q TOC Plus system. 8.2 Standards 8.2.1 Prepare PFOS standards for the standard curve. 3M Environmental Laboratory FACT-M- 1 .O Extraction of PFOS from Liver Page 3 of 8 Page 108 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 8.2.2 8.2.3 8.2.4 8.2.5 8.2.6 Weigh approximately 100 mg of PFOS into a 100 mL volumetric flask and record the actual weight. Bring to volume with methanol for a stock standard of approximately 1000 ppm (CLdmL). Dilute the stock solution with methanol for a working standard 1 solution of approxi.mately 50 ppm. Dilute the stock solution with methanol for a working standard 2 solution of approx. 5.0 ppm. Dilute the stock solution with methanol for a working standard 3 solution of approx. 0.50 ppm. 9.0 SAMPLEHANDLING 9.1 All livers are received frozen and must be kept frozen until the extraction is performed. 10.0 QUALITYCONTROL 10.1 Matrix Spikes 10.1.1 Prepare and analyze matrix spike and matrix spike duplicate samples to determine the accuracy of the extraction. 10.1.2 Prepare each spike using liver chosen by the analyst, usually a control liver. 10.1.3 Expected concentrations will fall in the mid-range of the initial calibration curve. 10.2 Continuing Calibration Checks 10.2.1 Prepare and analyze continuing calibration check samples to determine the continued linearity of the initial calibration curve. 10.2.2 One check is prepared per group of ten samples. For example, if a sample set = 34, four checks are prepared and extracted. 10.2.3 Prepare each continuing calibration check fiom the same liver homogenate used to prep the initial curve. 10.2.4 The expected concentration will fall within the mid-range of the initial calibration curve. 11.0 CALIBRATIOANND STANDARDIZATION 11.1 Prepare Liver Homogenate to Use for Standards 11.1.1 Weigh approximately 40 g of liver into a 250 mL Nalgene bottle containing 200 mLs Milli-QTMwater. Grind to a homogeneous solution. 11.1.2 If 40 g is not available, use appropriate amounts of liver and water in keeping with a 1:5 ratio. 11.1.3 See section 13.0 to calculate the actual density of liver. 3M Environmental Laboratory FACT-M-1 .O Extraction of PFOS from Liver Page 4 of 8 Page 109 3M Medical Department Study: T6316.1 Y Analytical Report: FACT-TOX-001 LRN-U2103 11.1.4 Add 1 mL of homogeneous solution to a 15 mL centrihge tube. Re-suspend homogeneous solution by shaking between aliquots while preparing a total of sixteen 1 mL aliquots of homogeneous solution in 15 mL centrifuge tubes. 11.1.5 Two 1 mL aliquots serve as matrix blanks. Use the standard concentrations and spiking amounts listed in table 1 to spike, in duplicate, two standard curves for a total of fourteen samples. Approximate Spiking Amounts for Calibration Standards (Approx. Conc.) - 0.50 ppm . 0.50 ppm 0.50 DDm 1 11 5.0 ppm 5.0 ppm 5.0 ppm 50 uDm - 4 20 40 10 . 20 30 4 PFOS in liver Blank 0.010 ppm 0.050 ppm 0.100 uum I- 0.250 ppm 0.500 ppm 0.750 ppm 1.OOO ppm 11.1.1 See section 13.0 to calculate actual concentrationsof PFOS in calibration standards. 11.2 Extract spiked liver homogenates following 12.14-12.24 of this method. Use these standards to establish each initial curve on the mass spectrometer. 12.0 PROCEDURES 12.1 Obtain frozen liver samples. In spent tissue, note that the liver has not been packaged with other tissues. 12.2 Cut approximately 1 g of liver using a dissecting scalpel. 12.3 Weigh the sample directly into a tared plastic sampule vial. 12.4 Record the liver weight in the study notebook. 12.5 Label the sampule vial with the study number, weight, liver ID, date and analyst initials. 12.6 Add 2.5 m L s of water to sampule vial. 12.7 Grind the sample. Put the grinder probe in the sample and grind for about 2 minutes, or until the sample is homogeneous. 12.8 Rinse the probe into the sample with 2.5 m L s water using a pipette. 12.9 Take the grinder apart and clean it with methanol after each sample. Follow AMDT-EP-22. 12.10 Cap the sample and vortex for 15 seconds. 3M Environmental Laboratory FACT-M- 1.O Extraction of PFOS from Liver Page 5 of 8 Page 110 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 12.11 Pipette 1 mL homogenate into a 15 mL polypropylene centrifuge tube. Label the centrifuge tube with the identical information as the sampule vial. (See Worksheet for documenting the remaining steps.) 12.12 Spike liver homogenates with the appropriate amount of PFOS standard as described in section 11.1 or Table 1. 12.13Pipette two 1 mL aliquots of Milli-Qm water to centrifuge tubes. These will serve as instrument blanks. 12.14Add 1 mL 0.5 M TBA and 2 mL of the 0.25 M sodium carbonatekodium bicarbonate buffer. 12.15Using a volumetric pipette, add 5 m L s ethyl acetate. 12.16 Cap each sample and put on the shaker for 20 minutes. 12.17 Centrifuge for 26 to 25 minutes, until layers are well separated. Set power on the centrifuge to approximately 3500 rpm. 12.18Remove 4 d s of organic layer, using a 5 mL graduated glass pipette, to a clean 15 mL centrifuge tube. Label this fresh tube with the same information as in 12.5. 12.19Put each sample on the analytical nitrogen evaporator until dry, approximately 2 to 3 hours. 12.20Add 1.OmL of methanol to each centrihge tube using a graduated pipette. 12.21 Vortex mix for 30 seconds. 12.22Attach a 0.2 pm nylon mesh filter to a 3 cc syringe and transfer the sample to this syringe. Filter into a 1.5 mL glass autovial. 12.23Label the autovial with the study number, animal number and gender, sample timepoint, matrix, final solvent, extraction date, and analyst(s) who performed the extraction. 12.24Cap and hold for electrospray mass spectrometry analysis. 12.25 Complete the worksheet and tape to page of study notebook. 13.0 DATAANALYSISAND CALCULATIONS 13.1 Calculations: 13.1.1 Calculate the density of liver (mg) in 1.OmL homogenate using the following equation: g of Liver x Average weight of ten 1 mL aliquots (mg) (g of Liver + g of Water) 3M Environmental Laboratory FACT-M- 1.O Extraction of PFOS from Liver Page 6 of 8 Page 111 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 13.1.2 Calculate actual concentrations of PFOS in calibration standards using the following equation: pL of Standard x Concentration (pg /mL) = Final Concentration (pg/g or mgkg) mg Liver'/ 1 mL homogenate of PFOS in Liver *Avefage weight of liver in solution as determined in 13.1.1, by weighing ten 1 mL homogenates of approximately 40 mg liver in 200 mL of Milli-Q water. 14.0 METHODPERFORMANCE 14.1 The method detection limit is equal to half the lowest standard in the calibration curve. - 15.0 POLLUTION PREVENTION AND WASTE MANAGEMENT 15.1 Sample waste is disposed in biohazard containers, flammable solvent waste is disposed in high BTU containers, and used glass pipette waste is disposed in broken glass containers located in the laboratory. 16.0 RECORDS 16.1 Complete the extraction worksheet and tape into the study notebook. 17.0 TABLESD, IAGRAMSF,LOWCHARTASN,D VALIDATION DATA 17.1 The validation report associated with this method is FACT-M-1.0 & 2.0-V-1, 18.0 REFERENCES 18.1 AMDT-EP-22, "Routine Maintenance of Ultra-Turrax T-25" 19.0 AFFECTEDOCUMENTS 19.1 FACT-M-2, "Analysis of Liver Extracts for Fluorochemicalsusing HPLC-Electrospray Mass Spectrometry" 20.0 REVISIONS Revision Number. Reason For Revision Revision Date 3M Environmental Laboratory FACT-M- 1.O Extraction of PFOS from Liver Page 7 of 8 Page 112 3M Medical Department Study: T6316.1 # Extraction Worksheet for FACT-M- 1 Analytical Report: FACT-TOX-001 LRN-U2103 Study # Sample Number Date and approx. 0.5 ppm approx. 5 ppm approx. 50 ppm Initials Liver Blank Remove a 4 mL aliquot of organic layer Put on Nitrogen Evaporator to dryness Evaporator Temperature Add 1.OmL of Methanol TN-A- Vortex 30 sec. Filter using a 3cc B-D syringe with a 0.2bm SRI filter into a 1.5 mL autosample vial MS/MSD/- Cont. Checks: Spiked uL of a ppm std ( ) for a final concentration of ppm. MS/MSD used sample . Cont. Checks used same homogenate as for std curve. 3M Environmental Laboratory FACT-M- 1.O Extraction of PFOS from Liver Page 8 of 8 Page 113 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 3M ENVIRONMENTLAALBORATORY METHOD EXTRACTIOONF POTASSIUM PERFLUOROOCTANESULFONATEOR OTHER ANIONIC FLUOROCHEMICASULRFACTANTSFROM SERUM FOR ANALYSIS USING HPLC-ELECTROSPRAYLMSAPSESCTROMETRY Method Number: FACT-M-3 -0 Author: Lisa Clemen Adoption Date: y txl q 8 Revision Date: NiA , Group Leader . Lk c. &#J Technical Reviewer Date Y/zz/93 Date zp E" ,a 1.0 SCOPEANDAPPLICATION 1.1 Scope: Thismethod is for the extraction of potassiumperfluorooctanesulfonate (PFOS)or other fluorochernical surfactants fiom serum. 1.2 Applicable Compounds: Fluorochemical surfactants or other fluorinated compounds. -1 + 3 1.3 Matrices: Rabbit, rat, and bovine serum or other sera as designated in the validation report. -.01. cha -J Microsoft 7.0.1/95 p> FACT-M-3.O Extraction of PFOS fiom Serum Page 1 of 8 3M Environmental Laboratory Page 114 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 2.0 SUMMAROYFMETHOD 2.1 This method describeshow to extract potassium peffluorooctanesulfonate(PFOS) or other anionic fluorochemical suffactants from serum using an ion pairing reagent and 5 .O mL of ethyl acetate. An ion pairing reagent is added to the sample and the analyte ion pair is partitioned into ethyl acetate. Four mL of extract are removed and put onto a nitrogen evaporatoruntil dry. Each extract is reconstituted in 1.O mL of methanol, then filtered through a 3 cc plastic syringe attached to a 0.2 pm nylon filter into glass autovials. 3.0 DEHNITIONS 3.1 None. 4.0 WARNINGASND CAUTIONS 4.1 Health and Safety Warnings: 4.1.1 Use universal precautions, especially laboratory coats, goggles, and gloves when handling animal serum, it may contah pathogens. 5.0 INTERFEE~ENCES 5.1 There are no known interferences at this time. 6.0 EQUIPMENT 6.1 The following equipment is used while carrying out this method. Equivalent equipment is acceptable. 6.1.1 Vortex mixer, VWR, Vortex Genie 2 6.1.2 Centrifige, Mistral 1000 or E C 6.1.3 Shaker, Eberbach or VWR 6.1.4 Nitrogen evaporator, Organomation 6.1.5 Balance, (It 0.100 gm) 7.0 SUPPLIES AND MATERIAIS 7.1 Gloves 7.2 Eppendorf or disposable pipettes 7.3 Nalgene bottles, capable of holding 250 mL and 1 L 7.4 Glass, type A, volumetric flasks 7.5 40 mL glass I-CHEM vials 7.6 Polypropylene centrifuge tubes, 15 mL 7.7 Labels 7.8 Syringes, capable of measuring 10 pL to 50 pL 7.9 Glass, type A, volumetric pipettes 7.10 Graduated pipettes 3M Environmental Laboratory FACT-M-3 .O Extraction of PFOS from Serum Page 2 of 8 Page 115 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 7.11 Electronic pipettor, Eppendorf or equivalent 7.12 Timer 7.13 Disposable plastic 3 cc syringes 7.14 Filters, nylon syringe filters, 0.2 pm, 25 mm 7.15 Crimp cap autovials Note: Prior to using glassware and bottles, rinse 3 times with methanol and 3 times with MilliQm water. Rinse syringes a minimum of 9 times with methanol, 3 rinses from 3 separate Vials. 8.0 REAGENTS AND STANDARDS 8.1 Reagents 8.1.1 Sodium hydroxide (J.TBaker or equivalent),(NaOH) 1ON: weigh approximately 200 grams NaOH. Pour into a 1000 mL beaker containing 500 liters (L) Milli-Qm water, mix until all solids are dissolved. Store in a 1 L Nalgene bottle. 8.1.2 Sodium hydroxide (J.TBaker or equivalent),(NaOH) 1N. Dilute 1ON 1:lO. Measure 10 mL of ION NaOH solutioninto a 100mL volumetric flask and dilute ' to volume using Milli-QTMwater. Storein a 125 mL Nalgene bottle. 8.13 Tetrabutylammonhmhydrogen sulfate (Kodak or equivalent), (TBA) OSM: Weigh approximately 169 grams ofTBA into a 1L volumetriccontaining 500 L Milli-Qm water. Adjust to pH 10using approximately 64mL of 1ON NaOH and dilute to volume with Milli-QTMwater. Add NaOH slowly while adding the last mL of NaOH because the pH changes abruptly. Store in a 1L Nalgene bottle. 8.1.3.1 TBA requires a check prior to each use to ensure pH = 10. Adjust as needed using 1N NaOH solution. 8.1.4 Sodium carbonate/sodium bicarbonate buffer (J.T. Baker or equivalent), (Na&03/NaHC03)0.25M:Weigh approximately26.5 g of sodium carbonate (NqCO,) and 21.O g of sodium bicarbonate (NaHCO,) into a 1L volumetric flask and bring to volume with Milli-Q"" water. Store in a 1 L nalgene bottle. 8.1.5 PFOS (3MSpecialty Chemical Division), molecular weight = 538. 8.1.6 Other fluorochemicals, as appropriate. 8.1.7 Ethyl Acetate, Omnisolv, glass distilled or HPLC grade. 8.1.8 Methanol, Omnisolv, glass distilled or HPLC grade. 811.9 Serum, frozen liquid from Sigma. 8.1.10 Control serum received with each sample set. 8.1.11 Milli-Qm water, all water used in this method should be Milli-Qm water and may be provided by a Milli-Q TOC Plus system. 3M Environmental Laboratory FACT-M-3 .O Extraction of PFOS from Serum Page 3 of 8 Page 116 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 8.2 Standards 8.2.1 Prepare PFOS standards for the standard curve. 8.2.2 Prepare other fluorochemical standards, as appropriate. 8.2.3 Weigh approximately 100 mg of PFOS into a 100 mL volumetric flask and record the actual weight. 8.2.4 Bring to volume with methanol for a stock standard of approximately 1000 ppm (I.Lg/mL)- 8.2.5 Dilute the stock solution with methanol for a working standard 1solution of approximately 50 ppm. 8.2.6 Dilute the stock solution with methanol for a working standard 2 solution of approx. 5.0 ppm. 8.2.7 Dilute the stock solution with methanol for a working standard 3 solution of approx. 0.50 ppm. 9.0 SAMPLJE~ANDLING 9.1 All sera are received frozen and must be kept frozen until the extractionis performed. 10.0 OUALITCYONTROL 10.1 Matrix Blanks and Method Blanks 10.1.1 Two 1.O mL aliquots of the serum are extracted following thisprocedure and used as matrix blanks. See section 11.1.2. 10.1.2 Two 1.0 mL aliquots of Milli-Qm water are extracted following this procedure and used as method blanks. 10.2 Matrix Spikes 10.2.1 Prepare and analyze matrix spike and matrix spike duplicate samples to determine the accuracy of the extraction. 10.2.2 Prepare each spike using serum chosen by the analyst, usually control serum received with each sample set. 10.2.3 Expected concentrationswill fall in the mid-range of the initial calibration curve. Additional spikes may be included and may fall in the low-range of the initial calibration curve. 10.3 Continuing Calibration Checks 10.3.1 Prepare and analyze continuing calibration check samples to determine the continued linearity of the initial calibration curve. 10.3.2 One check is prepared per group of ten samples. For example, if a sample set = 34, four checks are prepared and extracted. 3M Environmental Laboratory FACT-M-3 .O Extraction of PFOS fiom Serum Page 4 of 8 Page 117 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 10.3.3 Prepare each continuing calibration check fiom the same serum used to prep the initial curve. 10.3.4 The expected concentration will fall withinthe mid-range of the initial calibration curve. 11.0 CALIBRATION AND STANDARDIZATION 11.1 Prepare Serum Standards 11.1.1 Transfer 1 mL of serum to a 15 mL centrifuge tube. 11.1.2 If the majority of serum sample volumes are less than 1.O mL, extract standards using serum volumes inthe standards equal to the serum volumes in samples. Do not extract below 0.50 mL of serum. Record the serum volume on the extraction sheet. 11.1.3 Mix or shake between aliquots while preparing a total of sixteen aliquots of serum in 15 mL centrifugetubes. . 11.1.4 Two 1 mL or appropriate aliquots serve as matrix blanks. Typically use the standard concentrations and spiking amounts listed in table 1to spike, in duplicate, two standard curves for a total of fourteen samples. 11.1.5 Refer to the validation report FACT-M-3.0-V-1 and FACT-M-4.0-V-1 which lists the working ranges for calibration curves. Working Standard - (Approx. Conc.) 0.500 ppm 5.00 ppm 5.00 ppm 5.00 ppm 50.0 ppm 50.0 m m PL Approx. final conc. of - PFOS in serum B- -lank 20 0.0 10ppm 5 0.025 ppm 10 0.050 ppm 20 0.100 ppm 5 0.250 ppm - 10 0.500 vpm 11.1.4 See section 13.0 to calculate actual concentrations of PFOS in calibration standards. 11.2 Extract spiked serum standards following 12.6-12.16 of this method. Use these standards to establish each initial curve on the mass spectrometer. 3M Environmental Laboratory FACT-M-3.0 Extraction of PFOS fiom Serum Page 5 of 8 Page 118 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 12.0 PROCEDURES . 12.1 Obtain frozen serum samples and allow to thaw. 12.2 Vortex mix for 15 seconds then remove 1.OmL, or appropriate volume to a 15 mL polypropylene centrifuge tube. 1 2 3 Return serum samplesto freezer after extraction amount has been removed. 12.4 Record the serum volume on the extraction worksheet. The final methanol volume will equal the initial serum volume. 12.5 Label the tube with the study number, serum ID, date and analyst initials. See attached worksheet for documenting the remaining steps. 12.6 Spike serum with the appropriate amount of PFOS standard as described in section 11.1 or Table I for the calibration curve standards. Also spike matrix spikes and continuing calibration standards. 12.7 Vortex mix the standard curve samples, matrix spike samples, and continuing calibration samples for 15 seconds. 12.8 Add 1 mL 0.5 M TBA and 2 mL of the 0.25 M sodium carbonatdsodium bicarbonate buffer. 12.9 Using a volumetric pipette, add 5 mL ethyl acetate. 12.10 Cap each sample and put on the shaker for 20 minutes. 12.11 Centrifugefor 20 to 25 minutes,until layers are well separated. Set power on the centrifuge to approximately 3500 rpm. 12.12 Transfer 4 mL of organic layer, using a 5 mL, graduated glass pipette, to a clean 15 mL centrifugetube: Label this freshtube with the same information as in 12.5. 12.13 Put each sampleon the analytical nitrogen evaporatoruntil dry,approximately 2 to 3 hours. 12.14 Add 1.OmL or appropriate volume of methanol to each centrifugetube using a graduated pipette. (This volume equals the initial volume of serum used for the extraction.) 12.15Vortex mix for 30 seconds. 12.16 Attach a 0.2 pmnylon mesh filterto a 3 cc syringe and transfer the sampleto this syringe. Filter into a 1.5 mL glass autovial. 12.17Label the autovial with the study number, animal number and gender, sampletimepoint, matrix, final solvent, extraction date, and analyst(s) who performed the extraction. 12.18Cap and hold for HPLC-electrospray/massspectrometryanalysis. Extracts may be stored at 4" C until analysis. 12.19 Complete the extraction worksheet, attached to this document, and tape to page of study notebook. 3M Environmental Laboratory FACT-M-3 .O Extraction of PFOS from Serum Page 6 of 8 Page 119 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 13.0 DATAANALYSIASND CALCULATIONS 13.1 Calculations: 13.1.1 Calculate actual concentrationsof PFOS, or other appropriate fluorochemical,in calibration standards using the following equation: mL of Standard x Concentration(UE /mL) = Final Concentration (pg/mL) m L of Standard +Initial Serum Volume (a)of PFOS in Serum 14.0 METHOD PERFORMANCE 14.1 The-methoddetection limit is equal to halfthe lowest standard in the calibration curve. 15.0 POLLUTIOPNREVENTIONAND WASTE MANAGEMENT 15.1 Samplewaste is disposed inbiohazard containers, flammablesolvent waste is disposed in high BTU containers, and used glass pipette waste is disposed in broken glass containers located in the laboratory. 16.0 RECORDS 16.1 Complete the extraction wwksheet attached to this method, and tape into the study notebook. 17.0 TABLESD,IAGRAMFSL. OWCHARTANSD. VALIDATION DATA 17.1 The validation report associated with this method is FACT-M-3.0 & 4.0-V-1. 18.0 REFERENCES 18.1 None 19.0 AFFECTEDDOCUMENTS 19.1 FACT-M-4, "Analysis of Serum E2dract.s for Fluorochemicals using HPLC-Electrospray Mass Spectrometry" 20.0 REVISIONS Revision Number. Reason For Revision Revision - Date 3M Environmental Laboratory FACT-M-3.0 Extraction of PFOS from Serum Page 7 of 8 Page 120 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 ExtractionWorksheet for FACT-M-3 Study # - Sample Number set # H20Blank Serum Blank PFOS approx. 0.5 ppm actual ppm #W PFOS approx. 5 ppm actual ppm #W - PFOS approx. 50 ppm actual ppm #W - Date and Initials for Std. or Comments I I - ppm. MSNSD used sample . Cont. Checks used same serum as for std curve. FACT-M-3 .O Extraction of PFOS from Serum Page 8 of 8 3M Environmental Laboratory Page 121 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 3M ENVIRONMENTALLABORATORY METHOD EXTRACTION OF POTASSIUM~RFLUOROOCTANESULFONATEOR OTHER FLUoRoCREMICAL COMPOUNDS FROM SERUM FOR ANALYSIS USINGHPLCELECTROSPRAYMSAPESCSTROMETRY Method Number: ETS-8-4.1 Adoption Date: 03/01/99 Author: Lisa Clemen,Glenn Langenburg Revision Date: Lc/a7@? Approved By: Laboratory Manag& Date Technical Reviewer olllaL/4 9 Date 1.0 SCOPEANDAPPLICATION Scope: This method is for the extraction of potassium perfluorooctanesulfonate (PFOS) 0 or other fluorochemical compounds fiom senun. Applicable compounds: Fluorochemical surfactants or other fluorinated compounds. Matrices: Rabbit, rat, bovine, monkey, and human serum or other fluids as designated in the validation report. Word 6/95 3M Environmental Laboratory ETS-8-4.1 Extraction of PFOS from Serum Page 1 of 14 Page 122 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 2.0 SUMMARY OF METHOD 2.1 This method describes the procedure for extracting potassium perfluorooctanesulfonate (PFOS)or other fluorochemical surfactants fromserum, or otherfluids, using an ion pairing reagent and methyl-fert-butyl ether (AWE). In this method, seven fluorochemicalswere extracted: PFOS, PFOSA, PFOSAA, EtFOSE-OH, PFOSEA, M556, and surrogate standard (see 3.0 Definitions). An ion pairing reagent is added to the sample and the analyte ion pair is partitioned into MtBE. The MtBE extract is removed and put onto a nitrogen evaporatoruntil dry. Each extract is reconstituted in 1.O mL of methanol, then filtered through a 3 cc plastic syringe attached to a 0.2 p m nylon filter into glass autovials. 2.2 These sample extracts are analyzed following method ETS-8-5.1 or other appropriate methods. 3.0 DEFINITIONS 3.1 PFOS:perfluorooctanesulfonate(anion of potassium salt) C,F,,SO,' 3.2 PFOSA.perfluorooctanesulfonylamideC,F,,SO,NH, 3.3 PFOSM. peffluorooctanesulfonylamido(ethy1)acetate C8F,,S0,N(CH2CH3)CH2CO; 3.4 EtFOSE-OH: 2(N-ethylperfluorooctane sulfonamide)-ethyl alcohol C~F,,S02N(CH,CH3)CH2CH20H 3.5 PFOSEA: perfluorooctane sulfonyl ethylamide C,F,,SO,N(CH,CH,)H 3.6 M556:C8F,,SO,N(H)(CH,COOH) 3.7 Surrogate standard: lH-lH-2H-2H perfluorooctane sulfonicacid 4.0 WARNINGS AND CAUTIONS 4.1 Health and safety warnings 4.1.1 Use Universalprecautions,especiallylaboratory coats, goggles, and gloves when handling animal tissue, which may contain pathogens. 5.0 INTERFERENCES 5.1 There are no intexfkences known at this time. 6.0 EQUIPMENT 6.1 The following equipment is used while performing this method. Equivalent equipment is acceptable. 6.1.1 Vortex mixer, VWR, Vortex Genie 2 6.1.2 Centrifbge,Mistral 1000or IEC 6.1.3 Shaker, Eberbach or VWR 3M Environmental Laboratory ETS-84.1 Extraction of PFOS from Serum Page 2 of 14 Page 123 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 6.1.4 Nitrogen evaporator, Organomation 6.1.5 Balance (zk 0.100 g) 7.0 SUPPLIES AND MATERIALS 7.1 Gloves 7.2 Eppendorf or disposable pipettes 7.3 Nalgene bottles, capable of holding 250 mL and 1L 7.4 Volumetric flasks, glass, type A 7.5 I-CHEM Vials, glass, 40 mL glass 7.6 Centrifbge tubes, polypropylene, 15 mL 7.7 Labels 7.8 Oxford Dispenser- 3.0 to 10.0mL 7.9 Syringes,capableofmeasuring 5 pL to 50 pL 7.10 Graduated pipettes 7.11 Syringes, disposable plastic, 3 cc 7.12 Syringe filters, nylon, 0.2 p,25 mm 7.13 Timer 7.14 Crimp cap autovials and caps 7.15 Crimpers Note: Prior to using glassware and bottles, rinse 3 times with methanol and 3 times with Milli-Q"" water. Rinse syringes a minimum of 9 times with methanol, 3 rinses h m 3 separate vials. 8.0 REAGENTS AND STANDARDS 8.1 Type I reagent grade water, Milli-Qm or equivalent; all water used in this method should be Milli-QWwater and may be provided by a Milli-Q TOC PlusTMsystem 8.2 Sodiumhydroxide (NaOH), J.T Baker or equivalent 8.3 Tetrabutylammonium hydrogen suifate(TBA), Kodak or equivalent 8.4 Sodium carbonate (N%CO,), J.T.Baker or equivalent 8.5 Sodiumbicarbonate (NaHCO,), J.T. Baker or equivalent 8.6 Methyl-T-Butyl Ether, Omnisolv, glass distilled or HPLC grade 8.7 Methanol, Omnisolv, glass distilled or HPLC grade 8.8 Serum or blood, frozen fiom supplier 8.9 Fluorochemical standards 8.9.1 PFOS (3M Specialty Chemical Division), molecular weight = 538 8.9.2 PFOSA (3M Specialty Chemical Division), molecular weight = 499 ETS-8-4.1 Extraction of PFOS from Serum , Page 3 of 14 3M Environmental Laboratory Page 124 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 8.9.3 PFOSAA (3M SpecialtyChemical Division), molecularweight = 585 8.9.4 EtFOSE-OH(3MSpecialty Chemical Division), molecular weight = 570 8.9.5 PFOSEA (3M Specialty Chemical Division), molecular weight = 527 8.9.6 M556 (3M SpecialtyChemicalDivision), molecular weight =557 8.9.7 Surrogate standard:4-H, perfluorooctane: sulfonic acid (1-H,l-H, 2-H,2-H CBF,3S0,Hm) olecular weight =428 8.9.8 Other fluorochemicals, as appropriate 8.10 Reagent preparation NOTE: When preparing larger volumes thanlisted in reagent, standard,or surrogate preparation, adjust accordingly. 8.10.1 10N sodium hydroxide (NaOH): Weigh approximately200 g NaOH. Pour into a 1000 mL beaker containing 500 mL MillS-Qw water, mix until all solids are dissolved. Store in a 1L Nalgene bottle. 8.10.2 1N sodium hydroxide (NaOH): Dilute IO N NaOH 1:lO. Measure 10mI,of 10 N NaOH solution into a 100mLvolumetric flask and dilute to volume using Milli-QTMwater. Store in a 125 mL Nalgene bottle. 8.10.3 ' 0.5M tetrabutylammoniumhydrogen sulfate (TBA): Weigh approximately 169g of TBA into a 1L volumetriccontaining 500 dMilli-Qm water. Adjust to pH 10using approximately44to 54 mLof 1~0N NaOH (While adding the last mL of NaOH, add slowly because the pH changes abruptly). Dilute to volume with Milli-Qm water. Store in a 1L Nalgene bottle. 8.10.3.1 TBA requires a checkprior to each use to ensure pH = 10. Adjust as needed using 1N NaOH solution. 8.10.4 0.25M sodium carbonatelsodium bicarbonate buffer (Na$0,/N&C03): Weigh approximately 26.5 g of sodium carbonate (NqCO,) and 21.Og of sodium bicarbonate (NaHCO,) into a 1L volumetric flask and bring to volume with Mlli- Qm water. Store in a 1L Nalgenebottle:. 8.11 Standards preparation 8.11.1 Prepare PFOS standards for the standard curve. 8.1 1.2 Prepare other fluorochemkal standards, as appropriate. Multicomponent fluorochemicalstandardsare acceptable(for example, one working standard solution containing 1.00 ppm PFOS, 1.02 ppm PFOSA, 0.987 ppm PFOSAA, and 1-10ppm EtFOSE-OH.) 8.11.3 Weigh approximately 100 mg of PFOS into a 100 mL volumetric flask and record the actual weight. 8.11.4 Bring to volume with methanol for a stock standard of approximately 1000ppm (Cldd). 8.11.5 Dilute the stock solutionWith methanol for a working standard 1solutionof approximately 50 ppm. 8.11.6 Dilute working standard 1 with methanol for a working st&dard 2 solution of approx. 5.0 ppm. mS-8-4.1 Extraction of PFOS f'rorn Serum Page 4.of 14 3M Environmental Laboratory Page 125 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 8.11.7 Dilute working standard 1 with methanol for a working standard 3 solution of approx. 0.50 ppm. 8.12 Surrogate stock standard preparation 8.12.1 Weigh approximately 50-60 mg of surrogate standard 1-H,l-H2,-H,2-H, C$,,SO,H into a 50 mLvolumetric flask and record the actual weight. 8.12.2 Bring to volume with methanol for a surrogate stock of approximately 1000-1200 PPm* 8.12.3 Prepare a m g a t e working standard. Transfer approximately 1 mL of surrogate stock to a 10 mL volumetric flask and biing to volume with methanol for a working standard of 100ppm. Record tlhe actual volume transferred. 9.0 SAMPLEHANDLING 9.1 All samples are received fiozen and must be kept h z e n until the extraction is performed. 9.2 Allow samples to thaw to room temperature prior to extraction. 10.0 OUALITCYONTROL 10.1 Solvent Blanks, Method blanks and matrix blanks 10.1.1 An aliquot of 1.0 mL methanol is used as a solvent blank. 10.1.2 Extract two 1.0 mL aliquots of Milli-QTdwater following thisprocedure and use as method blanks. 10.1.3 Extract two 1.0 mL aliquots of the s e m i followingthisprocedure and use as matrix blanks. See 11.1.4. 10.2 Matrix spikes 10.2.1 Prepare and analyze matrix spike and mlatrixspike duplicate samples to determine the accuracy of the extraction. 10.2.2 Prepare each spike using a sample chosen by the analyst, usually the control matrix received with each sample set. 10.2.3 Expected concentrationswill fall in the ]mid-rangeof the initial calibrationcurve. Additional spikes may be included and may fall in the low-range of the initial calibration curve. 10.2.4 Prepare one matrix spike and matrix spike duplicate per 40 samples, with a minimum of 2 matrix spikes per batch. 10.3 Continuing calibration checks 10.3.1 Prepare continuing calibration check samples to ensure the accuracy of the initial calibration curve. 10.3.2 Prepare, at a minimum, one continuing check per group of 10 samples. For example, if a sample set = 34,four checks are prepared and extracted. 10.3.3 Prepare each continuing calibration check from the same matrix used to prepare the initial curve. 3M Environmental Laboratory ETS-8-4.1 Extraction ofPFOS from Serum Page 5 of 14 Page 126 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 10.3.4 The expected concentrations will fall within the mid-range of the initial calibration curve. Additional spikes mait be included that fall in the low-range of the initial calibration curve. This is necessary if the analyst must quantitate using only the low end of the calibrationcurve (for example, 5 ppb - 100ppb, rather than 5 ppb - 1000 ppb). 11.o CALIBRATION AND STANDARDIZATION 11.1 Prepare matrix calibration standards 11.1.1 Transfer 1 mL of serum to a 15mL centrifuge tube. 11.1.2 Ifmost sample volumes are less than 1.01 mL, extract standardswithmatrix volumes equal to the sample volumes. Do not extract less than 0.50 mL of matrix. Record each sample volume on the extraction sheet. 11.1.3 While preparing a total of twenty aliquolsin 15mL centrifuge tubes, mix or shake between aliquots. 11.1.4 Two 1 mL aliquots, or othex appropriatevolume, serve as matrix blanks. Typically use the standard concentrations and spiking amounts listed in Table 1, at the end of this section,to spike, in duplicate, two standard curves, for a total of eighteen standards, two matrix blanks, and two method blanks. 11.1.5 Refer to validation report ETS-8-4.0 & IETS-8-5.0-V-1,which lists the working ranges and the Linear CalibrationRange (LCR) for calibrationcurves. 11.1.6 Use Attachment D as an aid hcalculating the concentrations of the working standards. See Section 13.0 to calculate actual concentrations of PFOS in calibration standards. 11.2 To each standard, blank, or continuing check, add appropriate amount of swrogate working standard for the concentrationto fall within the calibration curve range 5 ppb - 1000 ppb. 113 Extract spiked matrix standardsfollowing 12.6-12.16 of this method. Use these standards to establish each initial curve on the mass spectrometer. 3M Environmental Laboratory ETS-8-4.1 ' Extraction of PFOS from Serum Page 6 of 14 Page 127 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Table 1 Approximate spiking amounts for standards and spikes Using 1.0 mL of maitrix . Working standard PL Approx. final conc. of (approx. conc.) analyte in matrix - Blank 0.500 ppm 10 0.005 ppm 0.500 ppm 20 0.010 ppm 5.00 ppm 5 0.025 ppm 5.00 ppm 10 0.050 ppm 5.00 ppm 20 0.100 ppm 50.0 ppm 5 0.250 ppm 50.0 ppm 10 0.500 ppm 50.0 ppm - 50.0 ppm 15 0.750 ppm 20 1.OOppm 12.0 PROCEDURE 12.1 Obtain fiozen samples and allow to thaw at room temperature or in a lukewarm waterbath. 12.2 Vortex mix for 15 seconds,then transfer 1.0 mL,or other appropriatevolume to a 15mL polypropylene centrifige tube. 12.3 Return unused samples to fieezer after extraction amounts have been removed, 12.4 Record the initial volume on the extractionworksheet. 12.5 Label the tube with the study number, sample E),date and analyst initials. See attached worksheet for documenting the remaining steps. 12.6 Spike all samples,includingblanks and standartis, ready for extraction with surrogate standard as described in 11.2. 12.7 Spike each matrix with the appropriateamount of standard as described in 11.1, or Table 1 in that section, for the calibration curve standards. Also prepare matrix spikes and continuing calibration standards. I 12.8 Vortex mix the standard curve samples, matrix spike samples, and continuing calibration samples for 15 seconds. 12.9 Check to ensure the 0.5 M TBA reagent is at pH[ 10. If not, adjust accordingly. 12.10 To each sample, add 1mL 0.5 M TBA and 2 rn12of 0.25M sodium carbonatdsodium bicarbonate buffer. 12.11 Using an Oxford Dispenser, add 5 mL methyl-twt-butyl ether. 12.12 Cap each sample and put on the shaker at a setting of 300 rpm,for 20 minutes. 12.13 Centrifuge for 20 to 25 minutes at a setting of 3500 rpm,or until layers are well separated. ETS-8-4.1 Extraction ofPFOS from Serum Page 7 of 14 3M Environmental Laboratory Page 128 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 12.14 Label a fiesh 15 mL centrifuge tube with the same information as in 12.5. 12.15 Remove 4.0mL of the organic layer to this clean 15mL centrihge tube. 12.16 Put each sample on the analytical nitrogen evaporator until dry, approximately i to 2 hours. 12.17 Add 1.O mL of methanol to each centrifugetuba using a graduatedpipette. 12.18 Vortex mix for 30 seconds. 12.19 Attach a 0.2 pm nylon mesh filterto a 3 cc syringe and transfer the sampleto this syringe. Filter into a 1.5 mL glass autovial or low-volume autovial when necessary. 12.20 Label the autovial with the study number, animal number and gender, sample timepoint, matrix, final solvent, extraction date, and analyst@)performing the extraction. 12.21 Cap and store extracts at room temperature or at approximately4 "C until analysis. 12.22 Complete the extractionworksheet, attached to this document, and tape in the study notebook or include in study binder, as appropriate. 13.0 DATA ANALYSIASN D CALCULATIONS 13.1 Calculations 13.1.1 Calculate actual concentrationsof PFOS,or other applicable fluomchemical, in calibration standards using the following equation: mL of standard x concentrationof standard lug /mU - . mL of standard +mL of surrogate standard +initialmatrix volume (d) Final Concentration (pg/mL,) of PFOS in matrix 14.0 METHODPERFORMANCE 14.1 The method detection limit (MDL) is analyte and matrix specific. Refer to MDL report for specificMDL and limit of quantitation (LOQ) values (see AttachmentsB and C). 14.2 The following quality control samples are extracted with each batch of samples to evaluate the quality of the extraction and analysis. 14.2.1 Method blanks and matrix blanks. 14.2.2 Matrix spike and matrix spike duplicate samples to determine accuracy and precision of the extraction. 14.2.3 Continuing calibration check samples to determine the continued accuracy of the initial calibrationcurve. 14.3 Refer to section 14 of ETS-8-5.1 for method performance criteria. 15.0 POLLUTION PREVENTION AND WASTE MANAGEMENT 15.1 Sample waste is disposed in biohazard containers, flammable solvent waste is disposed in high BTU containers, and used glass pipette waste is disposed in broken glass containers located in the laboratory. ETS-8-4.1 ExtractionofPFOS fiornSerum Page 8 of 14 3M Environmental Laboratory Page 129 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 16.0 RECORDS 16.1 Complete the extraction worksheet attached to this method, and tape in the study notebook or include in the 3-ring study binder, as appropriate. 17.0 ATTACHMENTS 17.1 Attachment A., Extraction worksheet 17.2 Attachment B, MDL/LOQ values and summary 17.3 Attachment C, Calibration standard concentration worksheet 18.0 REFERENCES 18.1 The validation report associated with this method is ETS-8-4.0& 5.0-V-1. 18.2 FACT-M-3.1,"Analysis ofSerum or Other Fluid Extracts for Fluorochemicals using HPLC-Electrospray Mass Spectrometry" 19.0 AFFECTED DOCUMENTS 19.1 ETS-8-5.1, "Analysis of Serum or Other Fluid Extracts for Fluorochemicals using HPLC-Electrospray Mass Spectrometry" 20.0 IREVISIONS . Revision Number 1 Reason For Revision Section 12.21 Changedto include sample storage at room temperatwe. Section 12.13 Added the shaker speed. Section 12.17 Final volume is 1.0 mL; not adjusted for initial volumes less than 1.0 mL. Revision - Date 04/02/99 3M Environmental Laboratory ETS-8-4.1 Extraction of PFOS from Serum Page 9 of 14 Page 130 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Extraction Worksheet ETS-8-4.I I 1 Shake 20 min. Shaker speed: Cont. Cal. Verificationsused same matrix as for std m e . Attachment A 3M Environmental Laboratory ETS-8-4.1 Extraction of PFOS fiom Serum I Page 10 of 14 Page 131 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Compound PFOS PFOSA PFOSAA EtFOSE-OH M556 PFOSEA MDL (ppb) 1.74 1.51 3.46 11.4 6.03 5.71 LOQ (ppb) 5.55 4.79 20.5 36.2 19.2 18.2 Linear Calibration Range (LCR) Approximate concentrations to be used for preparing the Standard Calibration Curve 5 ppb - 1000 ppb - 5 ppb 1000 ppb - 5 ppb 1000ppb 5 ppb - 1000 ppb - 5 ppb 1000ppb - 5 ppb 1000ppb Attachment B: MDWLOQ Summary 3M Environmental Laboratory ETS-8-4.1 ExtractionofPFOS from Serum Page 11 of 14 Page 132 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Prepared range LCR from % 1 1 ,I 11i- Rabbit Serum ofstandards bPb) (ng/mL) Full Range I [lCke 0.995 - 978 4.94-248 curve (Kpb) (ndmL) 24.8 - 978 4.94-248 83-108 4.67-1 1.0 I I W;;; 5.34-12.0 High curve 97.8 - 9713 0.995 - 978 97.8 - 978 4.94 - 978 85-106 4.849.80 4.60-10.5 Rabbit Serum Prepared range of standards (PPb) WmL) FullRange . 0.993 -976 LCB from CurYe (PPb) (ndmL) - 4.93 976 Low h e I I Highcurve I 1 l/x 4.93 - 97.6 24.8-976 0.993-976 4.93 - 97.6 1 24.8-978 I 4.93-976 % Recovery Range 88-103 87-105 I I 93-102 I 1 94-103 RSD Range 5.1 0-14.7 9.85-14.7 I 5.08-13.9 I 5.10-14.5 Rabbit Serum Full Range Prepared range of standards (PPb) (nl4m.L) - 0.991 974 LCR from curve 0 (ndmL) 24.7 - 974 YORecovery Range 81-111 RSD Range 4.18-10.6 Low Curve 4.92 - 247 9.74 - 247 97-107 6.38-2 1.8 High curve 49.2 - 974 97.4 - 974 85- 108 4.33-12.5 1/X I - - 0.991 974 9.74 974 95-1 15 4.1 1-23.2 I I I I I Attachment B: MDLnOQ Summary 3M Environmental Laboratory ETS-8-4.1 Extraction of PFOS from Serum 1 Page 12 of 14 Page 133 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Rabbit Serum Full Range Low Curve High curve 1/X Prepared range of standards (Ppb) (ng/mL) 0.993 - 976 4.93 - 97.6 49.3 - 976 0.993 - 493 LCR from curve (PPb) (ng/mL) 49.3 - 976 9.76 - 97.6 97.6 - 976 9.76 - 976 % Recovery Range 77-110 97-107 90-109 86-111 RSD Range 11.2-25.5 14.1-21.3 11.5-1 9.6 11.1-21.2 Rabbit Serum Prepared range of standards (PPW (ng/mL) Full Range Low Curve High curve 1/X 0.993 - 976 4.93 - 248 49.3 - 976 0.993 - 976 LCR fiom curve 0 (ndrnL) 2 4 8 - 976 9.76 - 248 49.3 - 976 9.76 - 976 % Recovery Range 96-106 91-110 86-106 95-1 17 RSD Range ~ 10.1- 16.2 11.8-19.5 10.2-18.2 10.1-19.1 Prepared range LCR fim YORecovery RSD Rabbit Serum of standards curve Range Range (PPb) ( n d d ) (PPb) (ng/mL) Full Range 0.993 - 976 24.8 - 976 88-106 4.82-17.9 Low Curve 1 4.93 ..97.6 1 9.76-97.6 1 100-105 1 5.95-18.2 I Highcurve I 97.6-976 1/X I 0.993-976 I 97.6-976 I I 9.76-976 I 81-111 97-110 I 5.11-9.74 1 I 4.77-19.5 I Attachment B:MDL/LOQ Summary 3M Environmental Laboratory ETS-8-4.1 ExtractionofPFOS from Serum Page 13 of 14 Page 134 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Ion Pair Standard Curves - Fluids Prep date(@: Standard number: Analyte(s): Equipment number: Sample matrix: Final solvent and TN: Blank fluiaidentifier: Method/revision: Target analyte(s): FC mix std approx. 0.500ppm: FC mix std approx. 5.00 ppm: FC mix std approx. 50.0ppm: Surrogate std approx 100 ppm: Actual concentrations of standards in the FC mix 493 500 524 494 501 513 735 746 782 737 749 766 976 989 1038 978 993 1017 Sernm Rabbit I PFOS 5.00-1000 PFOSA PFOSAA EtFOSE-OH PFOSEA M556 5.00-1000 I 5.00-1000 I 5.00-1000 I 5.00-1000 1 5.00-1000 Bovine Estimates only. Use values for rabbit. Rat Estimates only. Use values for rabbit. Monkey & Plasma Estimates only. Use values for rabbit. Human Estimates only. Use values for rabbit. Attachment C:Ion Pair StandardCurves ETS-8-4.1 Extraction of PFOS from Serum 3M Environmental Laboratory Page 14 of 14 Page 135 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 3M ENVIRONMENTLAALBORATORY METHOD EXTRACTIOONF POTASSIUM PERFLUOROOCTANESULFONATEOR OTHER FLUOROCHEMICCAOMLPOUNDS FROM LIVERFOR ANALYSIS USING HPLC- ELECTROSPRAY/MASPSESCTROMETRY Method Number: ETS-8-6.0 Author: Lisa Clemen, Robert Wynne Adoption Date: 03J 2 I"y Revision Date: $k Group Leader FWlg3 Date , $- @ Technical Reviewer 0 . 4 ,CI /as Date + 1.0 SCOPE AND APPLICATION 1.1 Scope: This method is for the extraction of potassium perfluorooctanesulfonate (PFOS) or TJ other fluorochemical compounds from liver. Y 0 1.2 Applicable Compounds: Fluorochemical surfactants or other fluorinated compounds. -+a 0 1.3 Matrices: Rabbit, rat, bovine, and monkey livers or other tissues as designated in the 2. validation report. -0 e. a Word 6.0195 ETS-8-6.0 Page 1 of 14 3M Environmental Laboratory Extraction of PFOS from Liver Page 136 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 2.0 SUMMARY OF METHOD 2.1 This method describes the procedure for extracting potassium perfluorooctanesulfonate (PFOS) or other fluorochemical surfactants from liver, or other tissues, using an ion pairing reagent and methyl-tert-butyl ether (MtBE). In this method, seven fluorochemicalscan be extracted: PFOS, PFOSA, PFOSAA, EtFOSE-OH,PFOSEA, M556, and surrogate standard. An ion pairing reagent is added to the sample and the analyte ion pair is partitioned into MBE. The MtBE extract is transferred to a centrifugetube and put onto a nitrogen evaporator until dry. Each extract is reconstituted in 1.O mL methanol then filtered through a 3 cc plastic syringe attached to a 0.2 pm nylon filter into glass autovials. 2.2 These sample extracts are analyzed following method ETS-8-7.0 or other appropriate methods. 3.0 DEFINITIONS 3.1 PFOS: perfluorooctanesulfonate (anion of potassium salt) C,FI7SO, 3.2 PFOSA: perfluorooctane sulfonylamide C,F,,SO,NH, 3.3 PFOSAA: perfluorooctane sulfonylamido (ethy1)acetate C,F,,SO,N(CH,CH,)CH,CO, 3.4 EtFOSE-OH: 2(N-ethylperfluorooctane su1fonamido)-ethyl alcohol C,F,,S0,N(CH,CH,)CH,CH20H 3.5 PFOSEA: perfluorooctane sulfonyl ethylamide C,F,,SO,N(CH,CH,)H 3.6 M556: C,F,,SO,N(H)(CH,COOH) 3.7 Surrogate standard: lH-lH-2H-2H perfluorooctane sulfonic acid 4.0 WARNINGS AND CAUTIONS 4.1 Health and Safety Warnings: 4.1.1 Use universal precautions, especially laboratory coats, goggles, and gloves when handling animal tissue, which may contain pathogens. 5.0 INTERFERENCES 5.1 There are no interferences known at this time. 6.0 EQUIPMENT 6.1 The following equipment is used while performing this method. Equivalent equipment is acceptable. 6.1.1 Ultra-Turrax T25 Grinder for grinding liver samples 6.1.2 Vortex mixer, VWR, Vortex Genie 2 6.1.3 Centrifuge, Mistral 1000 or IEC 6.1.4 Shaker, Eberbach or VWR 3M Environmental Laboratory ETS-8-6.0 Extraction of PFOS from Liver Page 2 of 14 Page 137 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 6.1.5 Nitrogen Evaporator, Organomation 6.1.6 Balance (sensitivity to 0.100 g) 7.0 SUPPLIES AND MATERIALS 7.1 Gloves 7.2 Dissecting scalpels 7.3 Eppendorf or disposable pipettes 7.4 Nalgene bottles, capable of holding 250 mL and 1 L 7.5 Volumetric flasks, glass, type A 7.6 I-CHEM vials, 40 mL glass 7.7 Plastic sampule vials, Wheaton, 6 mL (or appropriate size) 7.8 Centrifuge tubes, polypropylene, 15 mL 7.9 Labels 7.10 Oxford Dispensor - 3.0 to 10.0 ml 7.11 Syringes, capable of measuring 5 pL to 50 pL 7.12 Graduated pipettes 7.13 Syringes, disposable plastic, 3 cc 7.14 Syringe filters, nylon, 0.2 pm, 25 mm 7.15 Timer 7.16 Crimp cap autovials and caps 7.17 Crimpers Note: Prior to using glassware and bottles, rinse 3 times with methanol and 3 times with Milli- Q" water. Rinse syringes a minimum of 9 times with methanol, 3 rinses from 3 separate vials. 8.0 REAGENTASND STANDARDS 8.1 Type I reagent grade water, Milli-QTMor equivalent; all water used in this method should be Milli-QTMwater and be provided by a Milli-Q TOC PlusTMsystem 8.2 Sodium hydroxide (NaOH), J.T Baker or equivalent 8.3 Tetrabutylammoniumhydrogen sulfate(TBA),Kodak or equivalent 8.4 Sodium carbonate (Na.$O,), J.T. Baker or equivalent 8.5 Sodium bicarbonate (NaHCO,), J.T. Baker or equivalent 8.6 Methyl-tert-butyl ether, Omnisolv, glass distilled or HPLC grade 8.7 Methanol, Omnisolv, glass distilled or HPLC grade 8.8 Liver, frozen from supplier 8.9 Dry ice from supplier 8.10 Fluorochemical standards 8.10.1 PFOS (3M Specialty Chemical Division), molecular weight = 538 3M Environmental Laboratory ETS-8-6.0 Extraction of PFOS from Liver Page 3 of 14 Page 138 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 8.10.2 PFOSA (3M Specialty Chemical Division), molecular weight = 499 8.10.3 PFOSAA (3M Specialty Chemical Division), molecular weight = 585 8.10.4 EtFOSE-OH (3M Specialty Chemical Division), molecular weight = 570 8.10.5 PFOSEA (3M Specialty Chemical Division), molecular weight = 527 8.10.6 M556 (3M Specialty Chemical Division), molecular weight = 557 8.10.7 Surrogate standard: 4-H, perfluorooctanesulfonic acid (1-H,1-H, 2-H, 2-H C,F,,SO,H) molecular weight = 428 8.10.8 Other fluorochemicals,as appropriate 8.11 Reagent preparation NOTE: When preparing larger volumes than listed in reagent, standard, or surrogate preparation, adjust accordingly. 8.11.1 10N sodium hydroxide (NaOH): Weigh approximately200 g NaOH. Pour into a 1000 mL beaker containing 500 mL Milli-Q'" water, mix until all solids are dissolved. Store in a 1L Nalgene bottle. 8.11.2 1 N sodium hydroxide (NaOH): Dilute 10N NaOH 1:10. Measure 10mL of 10N NaOH solution into a 100mL volumetric flask and dilute to volume using Milli-QTMwater. Store in a 125mL Nalgene bottle. 8.11.3 0.5 M tetrabutylammoniumhydrogen sulfate (TBA): Weigh approximately 169 g of TBA into a 1 L volumetric containing 500 mL Milli-QTMwater. Adjust to pH 10 using approximately44 to 54 mL of 10N NaOH (While adding the last mL of NaOH, add slowly because the pH changes abruptly). Dilute to volume with Milli-QTMwater. Store in a 1L Nalgene bottle. 8.11.3.1 TBA requires a check prior to each use to ensure pH = 10. Adjust as needed using 1N NaOH solution. 8.11.4 0.25 M sodium carbonatehodium bicarbonate buffer (Na$O,/NaHCO,): Weigh approximately 26.5 g of sodium carbonate (N%COJ and 21.Og of sodium bicarbonate (NaHCO,) into a 1 L volumetric flask and bring to volume with Milli- QTMwater. Store in a 1 L Nalgene bottle. 8.12 Standards preparation 8.12.1 Prepare PFOS standards for the standard curve. 8.12.2 Prepare other fluorochemical standards, as appropriate. Multicomponent fluorochemical standards are acceptable (for example, one working standard solution containing 1.OO ppm PFOS, 1.02ppm PFOSA, 0.987 ppm PFOSAA, and 1.10 ppm EtFOSE-OH.) 8.12.3 Weigh approximately 100mg of PFOS into a 100mL volumetric flask and record the actual weight. 8.12.4 Bring to volume with methanol for a stock standard of approximately 1000ppm WmL). 8.12.5 Dilute the stock solution with methanol for a working standard 1 solution of approximately 50 ppm. 3M Environmental Laboratory ETS-8-6.0 Extraction of PFOS from Liver Page 4 of 14 Page 139 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 8.12.6 Dilute the stock solution with methanol for a working standard 2 solution of approx. 5.0 pprn. 8.12.7 Dilute the stock solution with methanol for a working standard 3 solution of approx. 0.50 ppm. 8.13 Surrogate stock standard preparation 8.13.1 Weigh approximately 50-60 mg of surrogate standard 1-H,l-H, 2-H, 2-H, C,F,,SO,H into a 50 ml volumetric flask and record the actual weight. 8.13.2 Bring to volume with methanol for a surrogate stock of approximately 1000-1200 PPm- 8.13.3 Prepare a surrogate working standard. Transfer approximately 1.O ml of surrogate stock to a 10 ml volumetric flask and bring to volurne with methanol for a working standard of 10-20 ppm. Record the actual volume transferred. 9.0 SAMPLHEANDLING 9.1 All samples are received fkozen and must be kept frozen until the extraction is performed. 10.0 QUALITY CONTROL 10.1 Matrix blanks and method blanks 10.1.1 An aliquot of 1.0 mL methanol is used as a solvent blank. 10.1.2 Extract two 1.O mL aliquots of Milli-Q"" water following this procedure and use as method blanks. 10.1.3 Extract two 1.0 mL aliquots of liver homogenate following this procedure and use as matrix blanks. Refer to 11.1.6. 10.2 Matrix spikes 10.2.1 Prepare and analyze matrix spike and matrix spike duplicate samples to determine the accuracy of the extraction. 10.2.2 Prepare each spike using a sample chosen by the analyst, usually a control liver received with each sample set. 10.2.3 Expected concentrationswill fall in the mid-range of the initial calibration curve. Additional spikes may be included and may fall in the low-range of the initial calibration curve. 10.2.4 Prepare one matrix spike and matrix spike duplicate per 40 samples, with a minimum of 2 matrix spikes per batch. 10.3 Continuing calibration verifications 10.3.1 Prepare continuing calibration verification samples to ensure the accuracy of the initial calibration curve. 10.3.2 Prepare, at a minimum, one continuing calibration verification sample per group of 10 samples. For example, if a sample set = 34, four verifications are prepared and extracted. 3M Environmental Laboratory ETS-8-6.0 Extraction of PFOS from Liver Page 5 of 14 Page 140 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 10.3.3 Prepare each continuing calibration verification from the same matrix used to prepare the initial curve. 10.3.4 The expected concentrationswill fall within the mid-range of the initial calibration curve. Additional spikes may be included that fall in the low-range of the initial calibration curve. This is necessary if the analyst must quantitate using only the low end of the calibration curve (for example, 5 ppb - 100 ppb, rather than 5 ppb - 1000 ppb). 11.0 CALIBRATION AND STANDARDIZATION 11.1 Prepare matrix calibration standards 11.1.1 Weigh approximately40 g of liver into a 250 mL Nalgene bottle containing 200 m L s Milli-QTMwater. Grind to a homogeneous solution, 11.1.2 If 40 g is not available, use appropriate amounts of liver and water to ensure a 1:5 ratio. 11.1.3 Refer to 13.0 to calculate the actual density of liver homogenate and the concentration of solid liver tissue dispersed in 1.0 mL of homogenate solution. 11.1.5 Add 1 mL of homogenate to a 15 mL centrihge tube. Re-suspend solution by shaking between aliquots while preparing a total of eighteen 1 mL aliquots of homogeneous solution in 15 mL centrifuge tubes. 11.1.6 Two 1mL aliquots, or other appropriate volume, serve as matrix blanks. 11.1.7 Typically use the standard concentrations and spiking amounts listed in Table 1, at the end of this section, to spike, in duplicate, two standard curves, for a total of eighteen samples, two matrix blanks, and two method blanks. 11.1.8 Refer to validation reports ETS-8-6.0 and ETS-8-7.0-V-1 or Attachment By which lists the working ranges and the Linear CalibrationRange (LCR) for calibration curves. 11.1.9 Use Attachment C as an aid in calculating the concentrations of the working standards. Refer to 13.0 to calculate actual concentrations of PFOS in calibration standards. 11.2 To each working standard, blank, or continuing verification, add appropriate amount of surrogate working standard for the concentration to fall within the calibration curve range 5 ppb - 1000ppb. 3M Environmental Laboratory ETS-8-6.0 Extraction of PFOS from Liver Page 6 of 14 Page 141 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 11.3 Extract spiked liver homogenates following 12.14-12.25 of this method. Use these standards to establish each initial curve on the mass spectrometer, Table 1 Approximate Spiking Amounts for Calibration Standards Working Standard 1.11 (Approx. Conc.) - - I 0.50 ppm I 0.50 ppm I 0.50 ppm I I 0.50 ppm I 2 I 4 I 10 I 20 I 0.50 ppm I 40 I 5.0 ppm I 10 I 5.0 ppm I 20 5.0 ppm 30 50 PPm 4 Approx. final conc. of PFOS in liver *- 0.500 ppm 12.0 PROCEDURE 12.1 Obtain frozen liver samples. 12.2 Cut approximately 1 g of liver using a dissecting scalpel. This part of the procedure is best performed quickly, not allowing the liver to thaw. 12.3 Weigh the sample directly into a tared plastic sampule vial. 12.4 Record the liver weight in'the study notebook. 12.5 Return unused liver portions to fieezer. 12.6 Add 2.5 mLs of water to sampule vial. 12.7 Grind the sample. Put the grinder probe in the sample and grind for about 2 minutes, or until the sample is homogeneous. 12.8 Rinse the probe into the sample with 2.5 mLs water using a pipette. 12.9 Take the grinder apart and clean it with methanol after each sample. Refer to AMDT-EP- 22. 12.10 Cap the sample and vortex for 15 seconds. Label the sampule vial with the study number, weight, liver ID, date and analyst initials. 3M Environmental Laboratory ETS-8-6.0 Extraction of PFOS from Liver Page 7 of 14 Page 142 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 12.11 Pipette 1.0 mL, or other appropriate volume, of homogenate into a 15 mL polypropylene centrifuge tube. Label the centrifuge tube with the identical information as the sampule vial. Refer to attached worksheet for documenting the remaining steps. 12.12 Pipette two 1 mL aliquots of Milli-QTMwater to centrihge tubes. These will serve as method blanks. 12.13 Spike all samples, including blanks and standards ready for extraction with surrogate standard as described in section 11.2. 12.14 Spike each matim with the appropriate amount of standard as described in 11.1, or Table 1 of that section, for the calibration curve standards. Also prepare matrix spikes and continuing calibration standards. 12.15 Vortex mix the standard curve samples, matrix spike samples, and continuing calibration samples for 15 seconds. 12.16 Check to ensure 0.5 M TBA reagent is at pH 10. If not, adjust accordingly. 12.17 To each sample, add 1 mL 0.5 M TBA and 2 mL of the 0.25 M sodium carbonate/sodium bicarbonate buffer. 12.18 Using an Oxford Dispenser, add 5 mL methyl-tert-butyl ether. 12.19 Cap each sample and put on the shaker at a setting of 300 rpm, for 20 minutes. 12.20 Centrifuge for 20 to 25 minutes at a setting of 3500 rpm, or until layers are well separated. 12.21 Label a fiesh 15 mL centrifuge tube with the same information as in 12.10. 12.22 Remove 4.0mL of the organic layer to the fresh 15 mL centrifuge tube. 12.23 Put each sample on the analytical nitrogen evaporatoruntil dry, approximately 1to 2 hours. 12.24 Add 1.O mL to each centrifuge tube using a graduated pipette. 12.25 Vortex mix for 30 seconds. 12.26 Attach a 0.2 pm nylon mesh filter to a 3 cc syringe and transfer the sample to this syringe. Filter into a 1.5 niL glass autovial or low-volume autovial when necessary. 12.27 Label the autovial with the study number, animal number and gender, sample timepoint, matrix, final solvent, extraction date, and analyst(s) performing the extraction. 12.28 Cap and store extracts at room temperature or at approximately4 "C until analysis. 12.29 Complete the extraction worksheet, attached to this document, and tape in study notebook or include in study binder, as appropriate. 3M Environmental Laboratory ETS-8-6.0 Extraction of PFOS from Liver Page 8 of 14 Page 143 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 13.0 DATAANALYSIASND CALCULATIONS 13.1 Calculations: 13.1.1 Calculate the average density of the liver homogenate by recording each mass of ten separate 1.O mL aliquots of homogenate. Average density (mg/mL) = Average mass (mg) of the aliauots 1.OmL aliquot 13.1.2 Calculate the amount of liver (mg) per 1.0 mL homogenate (or concentration of dispersed solid tissue per mL of homogenate suspension) using the following equation: g of Liver x Average density* of homopenate (mdmL) (g of Liver + g of Water) * refer to 13.1.1 for details. 13.1.3 Calculate actual concentrations of PFOS and other fluorochemicals in calibration standards using the following equation: pL of Standard x Concentration (LE /mL] = Final Concentration (&g or mg/kg) mg Liver / 1 mL homogenate* of PFOS in Liver *refer to 13.1.2 for details. 14.0 METHODPERFORMANCE 14.1 The method detection limit (MDL) is analyte and matrix specific. Refer to MDL report for specific MDL and limit of quantitation (LOQ) values (refer to Attachments B and C). 14.2 The following quality control samples are extractedwith each batch of samples to evaluate the quality of the extraction and analysis. 14.2.1 Method blanks and matrix blanks. 14.2.2 Matrix spike and matrix spike duplicate samples to determine accuracy and . precision of the extraction. 14.2.3 Continuing calibration verification samples to determine the continued accuracy of the initial calibration curve. 14.3 Refer to section 14 of ETS-8-7.0 for method performance criteria. 15.0 POLLUTION PREVENTION AND WASTE MANAGEMENT 15.1 Sample waste is disposed in biohazard containers, flammable solvent waste is disposed in high BTU containers, and used glass pipette waste is disposed in broken glass containers located in the laboratory. 3M Environmental Laboratory ETS -8-6.0 Extraction of PFOS from Liver Page 9 of 14 Page 144 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 16.0 RECORDS 16.1 Complete the extraction worksheet attached to this method, and tape in the study notebook or include in the 3-ring study binder, as appropriate. 17.0 TABLEDS,IAGRAMFSL, OWCHARTASN.D VALIDATION DATA 17.1 Attachment A, Extraction worksheet 17.2 Attachment B, MDWLOQ values and sumniary 17.3 Attachment C, Calibration standard calculation and concentrationworksheet 18.0 REFERENCES 18.1 The validation report associated with this method is ETS-8-6.0 & 7.0-V-1. 18.2 AMDT-EP-22, "Routine Maintenance of Ultra-Turrax T-25" 18.3 FACT-M-1.1, "Extraction of PFOS or Other Anionic Fluorochemical Surfactantsfiom ' Liver for Analysis Using HPLC-ElectrospraylMassSpectrometry" 19.0 AFFECTEDOCUMENTS 19.1 ETS-8-7.0, "Analysis of Liver Extracts for Fluorochemicalsusing HPLC-Electrospray Mass Spectrometry" 20.0 REVISIONS Revision Number. Reason For Revision Revision - Date 3M Environmental Laboratory ETS-8-6.0 Extraction of PFOS from Liver Page 10 of 14 Page 145 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Study # Matrix Box # way Date Sr>iked/Analvst . I ccv MS ~~ MSD Surrogate Std approx. pprn actual ppm # FC Mix Std FC Mix Std approx. 50 ppm actual ppm # Comments I I I I I I I I I I I - I I - - I - I I I I I I Cont. Cal. Verifications used the same matrix as for the standard curve. Attachment B: MDL/LOQ Values 3M Environmental Laboratory ETS-8-6.0 Extraction of PFOS from Liver Page 11 of 16 Page 146 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Compound PFOS PFOSA PFOSAA EtF'OSE-OH M556 PFOSEA MDL (ppb) 8.45 3.50 24.6 108 82.3 33.9 LOQ (ppb) 26.9 11.1 78.3 345 262 108 Linear Calibration Range (LCR) Approximate concentrations to be used for preparing the Standard Calibration Curve 30 ppb - 1200 ppb 12 ppb - 1200ppb 30 ppb - 1200 ppb 60ppb-900ppb" 60 ppb - 1200 ppb 30 ppb- 1200ppb MDWLOQ values in rat, bovine, and monkey liver were not statisticallydetermined. Two curves in each of these matrices were extracted and analyzed with the rabbit liver curves to determine equivalence. Responses in the rat, bovine, and monkey liver curves were equivalent to the rabbit responses, therefore, their MDL and LOQ will be assumed to be equivalent to those values as determined for the rabbit liver. Refer to LOQ Summary and MDL study in ETS-8-6.0 & 7.0-V-1for further information * EtFOSE-OH estimates only for MDL and LOQ. Did not meet criteria for validation. Compound: PFOS ComDound: PFOSA Rabbit 12- 1200 12- 1200. 12- 300 12- 300 60 - 1200 60 - 1200 Liver matrix Rabbit Prepared ' Range of range of average standards curve (ppb) (ng/mL) (ppb) (ng/mL) 6.16 - 1232 12 - 1200 L LCR from ave curve ( P P ~ )(ndmL) 30 - 1200 Range of low std curve ( P P ~ )(ng/mL) 30 - 900 LCR from low std curve (ppb) W m L ) 60 - 900 Range of high std curve (ppb) (ndmL) N/A LCR from high std curve (ppb) (ndmL) N/A Attachment B: MDL/LOQ Values 3M Environmental Laboratory ETS-8-6.0 Extraction of PFOS from Liver Page 12 of 16 Page 147 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Liver matrix Rabbit Prepared range of standards (ppb) (ng/mL) 6.17- 1235 Range of average curve (ppb) (ng/mL) 31 -900 LCR from ave curve (Ppb) (ndml-1 31 - 900 Range of low std curve ( P P ~ )(ng/mL) NIA LCR from low std curve ( P P ~ ()ng/mL) NIA Range of high std curve (ppb) (ndmL) NIA LCR from high std curve (ppb) (ng/mL) NIA Liver matrix Rabbit Prepared ' Range of range of average standards curve (ppb) W m L ) (ppb) (ng/mL) 6.17 - 1235 31 - 1200 LCR from ave curve (PPW (ng/mL) 31 - 1200 Range of low std CUNe (PPb) (ng/mL) N/A LCR from low std curve ( P P ~ )W m L ) NIA Range of high std curve (ppb) (ng/mL) NIA LCR from high std curve - (ppb) (ndmJ-1 NIA Compound: M556 Attachment C: Standard Calculations 3M Environmental Laboratory ETS-8-6.0 Extraction of PFOS from Liver Page 13 of 14 Page 148 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Ion Pair Standard Curves - Tissue Prep date(s): Analytets): Sample matrix: Methodhevision: Target analyte(s): FC mix std approx. 0.500 ppm: FC mix std approx. 5.00 ppm: FC mix std approx. 50.0ppm: Surrogate std approx. 100 ppm: Standard number: Equipment number: Final solvent and TN: Blank livedidentifier: Actual concentrations of standards in the FC mix PFOS PFOSA PFOSAA EtFOSE PFOSEA Final Final Final conc Final Final I I I I conc conc ndg conc conc c onc I n g k np/g ngk +'* 5- ..9-9. I 5.99 I 5.99 I 5.99 I . n5g.9/9g 5.99 1 12.0 I - 12.0 I I 12.0 I 1 12.0 1I 12.0 I 12.0 II 29.9 29.9 29.9 29.9 29.9 2!9.9 59.9 59.9 59.9 59.9 59.9 59.9 120 120 120 120 120 120 299 299 299 299 299 299 599 599 599 599 599 599 1 I 1 I I 1 8-9.8- I 898 I 898 9 898 1198 1198 1198 1198 898 1198 898 1198 Surrogate Std conc ng/mL 100 Surrogate Final conc ng/mL 0.500 All Am't spiked mL 0.005 Attaclment C: Standard Calculations 3M Environmental Laboratory ETS-8-6.0 Extraction of PFOS from Liver Page 14 of 14 Page 149 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 3M ENVIRONMENTALLABORATORY METHOD ANALYSIS OF FLUOROCHEMICAINLLSIVER-EXTRACUTSSING HPLC-ELECTROSPRAYAMSAPSESCTROMETRY Method Number: FACT-M-2.0 - Author: Lisa Clemen Approved By: Qql'$%- Laboratory Manager Adoption Date: s / J b198 Revision Date: / d / A Date & & A CQMhLk Technical Reviewer d 27198 Date SCOPE AND APPLICATION 13 Scope: This method is for the analysis of extracts of liver or other tissues for fluorochemical *surfactants using HPLC-electrospray/mass spectrometry. 0 Q Applicable Compounds: Potassium perfluorooctanesulfonate, anionic fluorochemical a surfactants, or other ionizable compounds. Matrices: Rabbit, rat, bovine, and monkey livers or other livers as designated in the 0validation report. 2. 32 &rd 7.0.1/95 FACT-M-2.0 Page 1 of 8 Analysis of Liver Extract Using ES/MS 3M Environmental Laboratory Page 150 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 2.0 SUMMARY OF METHOD 2.1 This method describes the analysis of fluorochemicalsurfactants extracted from liver using HPLC-electrospray/mass spectrometry.The analysis is performed by monitoring a single ion characteristic of a particular fluorochemical, such as the potassium perfluorooctanesulfonate(PFOS) anion, M/Z= 499. Samples may also be screened to veri@ compound identification. 3.0 DEFINITIONS 3.1 None. 4.0 WARNINGS AND CAUTIONS 4.1 Health and Safety Warnings: 4.1.1 Use caution with the voltage cable for the probe. When the voltage cable is plugged into the probe DO NOT TOUCH THE PROE$E,there is risk of electrical shock. 4.2 Cautions: 4.2.1 Do not run solvent pumps above capacity of ,400bar (5800 psi). If pressure goes over 400 bar, the HP1100will initiate autom;3ticshutdown. 4.2.2 Do not run solvent pumps to dryness. 5.0 INTERFERENCES 5.1 Teflon should not be used for sample storage or any part of instrumentation that comes in contact with the sample or extract. 6.0 EQUIPMENT 6.1 Equipment listed below may be changed in order to optimize the system. 6.1.1 Micromass Electrospray Mass Spectrometer 6.1.2 HP1100 low pulse solvent pumping system and autosampler. 7.0 SUPPLIES AND MATERIALS 7.1 Supplies 7.1.1 Nitrogen gas, refrigerated liquid, regulated to approximately 100 psi. 7.1.2 HPLC column, specifics to be determined by the analyst. 7.1.3 Capped autovials or capped 15 mL, centrifuge tubes. 8.0 REAGENTSAND STANDARDS 8.1 Reagents 8.1.1 Methanol, HPLC grade or equivalent. Word 7.0.1/95 3M Environmental Laboratory FACT-M-2.O Analysis of Liver Extract Using ESMS Page 2 of 8 Page 151 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 8.1.2 Milli-QTMwater, all water used in this method should be Milli-QTMwater and may be provided by a Milli-Q TOC Plus system. 8.1.3 Ammonium acetate, HPLC grade or equivalrmt. 8.2 Standards 8.2.1 Typically one H,O blank, one liver blank, and seven liver standards are prepared during the extraction procedure. See FACT-M-1. - 9.0 SAMPLHEANDLING 9.1 Fresh liver standards are prepared with each analysis. Extracted standards and samples are stored in capped autovials or capped 15 mL centrifuge tubes until analysis. 9.2 If analysis will be delayed, extracted standards and samples may be refrigerated until analysis can be p'erformed. 10.0 QUALITYCONTROL 10.1 Matrix Blanks and Method Blanks 10.1.1 Analyze a method blank and matrix blank prior to each calibration curve. 10.2 Matrix Spikes 10.2.1 Analyze a matrix spike and matrix spike duplicate with each analysis. 10.2.2 Expected concentrationswill fall in the mid-range of the initial calibration curve. Additional spike concentrations may fall in the low-range of the initial calibration curve. 10.2.3 See section 13 to calculate percent recovery. 10.3 Continuing Calibration Checks 10.3.1 Analyze a mid-range calibration standard afber every tenth sample. If a significant change (*30%) in peak area occurs, relative to the initial standard curve, stop the run.Only those samples analyzed before the last acceptable calibration standard will be used. The remaining samples must be reanalyzed. 10.3.2 See section 13 to calculate percent difference. 10.4 System Suitability 10.4.1 System suitability (e.g. peak area, retention time and peak shape, etc.) will be assessed for each run. 11.o CALIBRATION AND STANDARDIZATION 11.1 Analyze the extracted liver standards prior to and following each set of extracts. The mean of two standard values, at each standard concentration, will be plotted by linear regression for the calibration curve using MassLynx or other suitable software. 3M Environmental Laboratory FACT-M-2.0 Analysis of Liver Extract Using EYMS Page 3 of 8 Page 152 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 11.2 The 8value for the data should be 0.98 or greater. Lower values may be acceptable at the discretion of the analyst. 11.3 If the curve does not meet requirements, perfom routine maintenance or reextract the standard curve (if necessary) and reanalyze. 12.0 PROCEDURES 12.1 Acquisition Sd up 12.1.1 Click on start button in the Acquisition Comrol Panel. Set up a sample list. Assign a filename using letter-MO-DAY-last digit of year-sample number, assign a method (MS) for acquiring, and type in sample descriptions. 12.1.2 To create a method click on scan button in the Acquisition control panel and select SIR. SetJonization Mode as appropriate anti mass to 499 or other appropriate masses. A scan is usually collected along with the SIRS. Save method. 12.1.3 Typically the sample list begins with the first set of liver standards and ends with the second set of standards. , 12.1.4 Samples are analyzed with a continuing calibration check injected after every tenth sample. Solvent blanks should be analyzed ]periodicallyto monitor possible analyte carryover and are not considered samples but may be included as such. 12.2 Using the Autosampler 12.2.1 Set up sample tray according to the sample 1ist prepared in section 12.1.1, 12.2.2 Set-up the HP11OO/autosampler at the followingconditions or at conditions the analyst considers appropriate for optimal response. Record actual conditions in the instrument logbook: 12.2.2.1 Sample size = 10 pL injection with a sample wash 12.2.2.2 Inject/sample = 1 12.2.2.3 Cycle time = 15 minutes 12.2.2.4 Solvent ramp = Time 2.0 mM Ammonium acetate 0.00 min. 45% 7.5 min. 90% 10% 11.O min. 90% 11.5 min. 45% Note: In this instrument configuration, the run must be set up on the electrospray software with a "Waiting for inlet start" message before the "Start" button is pressed on the HP Workstation. 12.2.2.5 Press the "Start" button. 3M Environmental Laboratory FACT-M-2.0 Analysis of Liver Extract Using ES/MS Page 4 of 8 Page 153 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 12.3 Instrument Sep-up 12.3.1 Refer to AMDT-EP-31 for more details. 12.3.2 Check the solvent level in reservoirs and refill1 if necessary. 12.3.3 Check the stainless steel capillary at the end of the probe. Use an eye piece to check the tip. The tip should be flat with no jagged edges. If the tip is found to be unsatishctory, disassemble the probe and replace the stainless steel capillary. 12.3.4 Set HPLC pump to "On". Set the flow to 10 .-500 uL/min or as appropriate. Observe droplets coming out of the tip of the probe. Allow to equilibrate for approximately 10 minutes. 12.3.5 Turn on the nitrogen. A fine mist should be expelled with no nitrogen leaking around the tip of the probe. 12.3.6 The instrknent uses these parameters at the following settings. These settings may change in order to optimize the response: 12.3.6.1 Drying gas 250-400 litershour ' 12.3.6.2 ESI nebulizing gas 10-15 liters/how 12.3.6.3 LC constant flow mode flow rate 10- 500 uL/min 12.3.6.4 Pressure <400 bar (This parameter is not set, it is a guide to ensure the instrument is operating correctly.) 12.3.7 Carefully guide the probe into the opening. Insert probe until it will not go any M e r . Connect the voltage cables to the probe. 12.3.8 Record tune parameters in the instrument log. 12.3.9 Using the cross-flow counter electrode in the ESMS source is recommended for the analysis of biological matrices. 12.3.10 Click on start button in the Acquisition Control Panel. Press the start button at top of sample list. Ensure start and end sample number includes all samples to be analyzed. 13.0 DATAANALYSIASND CALCULATIONS 13.1 Calculations: 13.1.1 Calculate matrix spike percent recoveries using the following equation: % Recovery = Observed Result - Background Result x 100 Expected Result 13.1.2 Calculate percent difference using the following equation: % Difference = Expected Conc. - Calculated Co- nc. x 100 Expected Conc. 3M Environmental Laboratory FACT-M-2.0 Analysis of Liver Extract Using ESMS Page 5 of 8 Page 154 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 13.1.3 Calculate actual concentration of PFOS anion in total liver (mg): ug PFOS anion calc. fiom std curve g of liver used for analysis lo00 ug/ 1 mg x `Total mass of liver (g) 14.0 METHODPEWORMANCE 14.1 The method detection limit is equal to at least three limes the baseline noise in the matrix blank. 14.2 The practical quantitation limit is equal to the lowest standard in the calibration curve. 15.0 POLLUTION PREYENTION AND WASTE MANAGEMENT 15.1 Sample waste is disposed in biohazard containers, flammable solvent waste is disposed in high BTU containers, and glass pipette waste is disposed in broken glass containers. All containers are located in the laboratory. , 16.0 RECORDS 16.1 Store chromatograms in the study folder. Each chromatogram should have the following information included either in the header or hand written on the chromatogram: study number, sample name, extraction date, and dilution factor (if applicable). 16.2 Plot calibration curve by linear regression and store in the study folder. 16.3 Print sample list from MassLynx and tape into the instrument runlog. 16.4 Print data integration summary fiom MassLynx and tape into the instrument runlog. 16.5 Copy instrument runlog pages, including instrument parameters and sample results, and tape into appropriate study notebook. 16.6 Summarize data using suitable software and store in the study folder. 16.7 Back up electronic data to appropriate media. Record in study notebook the file name and location of backup electronic data. 17.0 TABLESD,IAGRAMSF,LOWCHARTASN,D VALIDAT1ON DATA 17.1 Attachment A: FACT-M-2 Data reporting spreadsheet 17.2 The validation report associated with this method is FACT-M-1.O& 2.0-V-1. 18.0 REFERENCES 18.1 AMDT-EP-31, "Operation of VG Platform Electrospray Mass Spectrometer" 3M Environmental Laboratory FACT-M-2.0 Analysis of Liver Extract Using ES/MS Page 6 of 8 Page 155 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 19.0 AFFECTEDOCUMENTS 19.1 FACT-M- 1 .O, "Extraction of Potassium Perfluorooctanesulfonate from Liver for Analysis Using HPLC-ElectrosprayNass Spectrometry" 20.0 REVISIONS Revision Number. Reason For Revision- Revision Date 3M Environmental Laboratory FACT-M-2.0 Analysis of Liver Extract Using ESMS Page 7 of 8 Page 156 3M Medical Department Study: T6316.1 L Analytical Report: FACT-TOX-001 LRN-U2103 Laboratory Study # Study: Test Material: . MatridFinal Solvent: MethodRevision: Analytical Equipment System Number: Instrument SoftwareNersion: Filename: R-Squared Value: Slope: Y Intercept: Date of Extractiodhalyst; Date of Analysis/Analyst: Group Dose Sample# , Concentration ug/mL - Initial Vol. mL Dilution Factor Final Conc. ug/mL Slope: Taken from linear regression equation. 3M Environmental Laboratory FACT-M-2.0 Analysis of Liver Extract Using ESMS Page 8 of 8 Page 157 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 3M ENVIRONMENTLAALBORATORY I METHOD ANALYSISOF F'LUOROCHEMICALS IN SERUM EXTFUCTSUSING HPLC-ELECTROSPRAY/MA!?S SPECTROMETRY Method Number: FACT-M-4.0 Author: Lisa Clemen Adoption Date: 91 q 8 Revision Date: NlA v i App-r.oved By: x - - - _ !d. Laboratory ManGer " Group Leader dk C L Technical Reviewer Date i Date Y/l4/98 Date m X f D a I 0, 1.1 Scope: Thismethod is for the analysis of extractsof serum or tissue for fluorochemical 0 surfactantsusing HPLC-electrospray/massspectrometry. h U 73 1.2 Applicable Compounds: Potassium perfluorooctanedowte, anionicfluorochemical 0 surfactants, or other ionizable compounds. 1.3 Matrices: Rabbit, rat, and bovine serum or other sem as designated in the validation report. - 3 Word 7.0.1/95 3M Environmental Laboratory FACT-M-4.0 Analysis of Serum Extract Using ESMS Page 1 of 8 Page 158 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 2.0 SUMMARY OF METHOD 2.1 This method describes the analysis of fluorochemical surfactants extracted from serum using HPLC-electrospray/massspectrometry. The (analysisis performed by monitoring a single ion characteristic of a particular fluorochemiical, such as the potassium perfluorooctanesulfonate (PFOS) anion, M/Z= 499. Samples may also be screened to verifl compound identification. 3.0 DEFINITIONS 3.1 None. 4.0 WARNINGSAND CAUTIONS 4.1 Health and Safety Warnings: 4.1.1 Use caution with the voltage cable for the probe. When the voltage cable is plugged into the probe DO NOT TOUCH THE PROBE, there is risk of electrical shock. 4.2 Cautions: 4.2.1 Do not run solventpumps above capacity o:F400 bar (5800 psi). If pressure goes over 400 bar, the HP1100 will initiate autornatic shutdown. 4.2.2 Do not run solvent pumps to dryness. 5.0 INTERFERENCES 5.1 Teflon should not be used for sample storage or any part of instrumentation that comes in contact with the sample or extract. 6.0 EQUIPMENT 6.1 Quipment listed below may be changed in order tcl optimize the system. 6.1.1 Micromass Electrospray Mass Spectrometer 6.1.2 HP1100 low pulse solvent pumping system and autosampler. 7.0 SUPPLIES AND MATERIALS 7.1 Supplies 7.1.1 Nitrogen gas, refiigerated liquid, regulated to approximately 100 psi. 7.1.2 HPLC column, specifics to be determined by the analyst. 7.1.3 Capped autovialsor capped 15mL centrifugetubes. 8.0 REAGENTS AND STANDARDS 8.1 Reagents 8.1.1 Methanol, HPLC grade or equivalent. 3M Environmental Laboratory FACT-M-4.0 Analysis of Serum Extract Using ESMS Page 2 of 8 Page 159 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 8.1.2 Milli-Qm water, all water used in this method should be Milli-Qm water and may be provided by a Milli-Q TOC Plus system. 8.1.3 Ammonium acetate, HPLC grade or equivsilent. 8.2 Standards 8.2.1 Typically one H20 blank, one serum blank, and seven serum standards are prepared during the extraction procedure. See FACT-M-3. 9.0 SAMPLEHANDLING 9.1 Fresh serum standards are prepared with each anal;ysis. Extracted standards and samples are stored in capped autovials or capped 15 mL, centrifuge tubes until analysis. 9.2 If analysis will be delayed, extracted standardsand samplesmay be refrigerated at 4 O C mtil analysis can be performed. 10.0 QUALITY CONTROL 10.1 Matrix Blanks and Method Blanks 10.1.1 Analyze a method blank and a matrix blank prior to each calibration curve. 10.2 Matrixspikes 10.2.1 Analyze a matrix spike and matrix spike dyplicate with each analysis. 10.2.2 Expected concentrations will fall in the mid-range of the initial calibration curve. Additional spike concentrationsmay fall in the low-range of the initial calibration curve. 10.2.3 See section 13 to calculatepercent recovery. 10.3 Continuing Calibration Checks (* 10.3.1 Analyze a mid-range calibration standard after every tenth sample. If a significant change 30%) in peak area occurs, relative to the initial standard curve, stop the run.Only those samples analyzed before the: last acceptable calibrationstandard will be used. The remaining samples must be reanalyzed. 10.3.2 See section 13 to calculate percent difference. 10.4 System Suitability 10.4.1 System suitability (e.g., peak area, retention time, peak shape, etc.) will be assessed for each run. 11.0 CALIBRATION AND STANDARDIZATION 11.1 Analyze the extracted serum standards prior to and allowing each set of extracts. The mean of two standard values, at each standard concentration,will be plotted by linear regression for the calibration curve using MassLp: or other suitablesoftware. 3M Environmental Laboratory FACT-M-4.0 Analysis of Serum Extract Using ESMS Page 3 of 8 Page 160 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 11.2 The 3 value for the data should be 0.98 or greater. Lower values may be acceptable at the discretion of the analyst. 11.3 If the curve does not meet requirements, perform routine maintenance or reextract the standard curve (if necessary) and reanalyze. 12.0 PROCEDURES 12.1 Acquisition Set up 12.1.1 Click on start button in the Acquisition Coritrol Panel. Set up a sample list. Assign a filename using letter-MO-DAY-last digit of year-sample number, assign a method (MS) for acquiring, and type in sample descriptions. 12.1.2 To create a method click on scan button in ithe Acquisition control panel and select SIR (Single Ion Recording). Set Ionization Mode as appropriate and mass to 499 or other appropriate masses. A scan is usually collected along with the SIRS. Save method. 12.1.3 Typically the sample list begins with the first set of serum standards and ends with the second set of standards. 12.1.4 Samples are analyzed with a continuing calibration check injected after every tenth sample. Solvent blanks should be analyzed periodically to monitor possible and* carryover and are not considered samples biut may be included as such. 12.2 Using the Autosampler 12.2.1 Set up sample tray according to the sample list prepared in section 12.1.1. 12.2.2 Set-up the HPl lOO/autosampler at the followingconditions or at conditionsthe analyst considersappropriatefor optimal response. Record actual conditions in the instrument logbook 12.2.2.1 Sample size = 10 pL injection with a sample wash 12.2.2.2 Inject/sample = 1 12.2.2.3 Cycle time = 15 minutes 12.2.2.4 Solvent ramp = Z - 7 1 El 11.Omin. I Ammonium acetate I 11.5min. I 45% I 55?h Note: In this instrument configuration, the nm must be set up on the electrospray software with a "Waiting for inlet start7' mesmge before the "Start7' button is pressed on the HP Workstation. 12.2.2.5 Press the "Start" button. FACT-M-4.0 Analysis of Serum Extract Using ESMS Page 4 of 8 3M Environmental Laboratory Page 161 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 12.3 Instrument Set-up 12.3.1 Refer to AMDT-EP-3 1 for more details. 12.3.2 Check the solvent level in reservoirs and refill if necessary. 12.3.3 12.3.4 Check the stainless steel capillary at the end of the probe. Use an eye piece to check the tip. The tip should be flat with no jagged edges. If the tip is found to be unsatisfactory, disassemble the probe and replace the stainless steel capillary. Set HPLC pump to "On". Set the flow to 10 - 500 ul/min or as appropriate. Observe droplets coming out of the tip of the probe. Allow to equilibrate for approximately 10 minutes. 12.3.5 Turn on the nitrogen. A fine mist should bt: expelled with no nitrogen leaking around the tip of the probe. 12.3.6 The instrument uses these parameters at the following settings. These settings may change in order to optimizethe response: 12.3.6.1 Drying gas 250-400 literdhour 12.3.6.2 ESI nebulizing gas 10-15 literdhoirr 12.3.63 HPLC constant flow mode flow rate 10 - 500 pL/min 12.3.6.4 Pressure <400 bar (This parameter is not set, it is a guide to ensure the HPLC is operating correctly.) 12.3.7 Carefully guide the probe into the opening. Insert probe until it will not go any further.Connect the voltage cablesto the probe. 12.3.8 Record tune parameters in the instrument log. 12.3.9 Using the cross-flowcounter electrode in the ESMS source is recommended for the analysis of biological matrices. 12.3.1OClick on start button in the Acquisition Control Panel. Press the start button at top of sample list. Ensure start and end sample number includes all samples to be analyzed. 13.0 DATAANALYSIASND CALCULATIONS 13.1 Calculations: 13.1.4 Calculate matrix spike percent recoveries using the following equation: % Recovery = - Observed Result Background Result x 100 Expected Result 13.1.5 Calculate percent difference using the following equation: YODifference = Emected Conc. - Calcul!atedConc. K 100 Expected Cclnc. 3M Environmental Laboratory FACT-M-4.0 Analysis of Serum Extract Using ESMS Page 5 of 8 Page 162 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 13.1.6 Calculate actual concentration of PFOS, or other fluorochemical, anion in serum (IZg/mL): pg of PFO calc. from std. Curve x Dilution Factor x Final Volume (mL) Initial Volume of serum (d) 14.0 METHODPERFORMANCE 14.1 The method detection limit is equal to half the lowlest standard in the calibration curve. 14.2 The practical quantitation limit is equal to the lowest standard in the calibration curve. 15.0 POLLUTION PREVENTION AND WASTE MANAGEMENT 15.1 Sample extract waste and flammable solvent is disposed in high BTU containers, and glass pipette waste is disposed in broken glass container:; located in the laboratory. 16.0 RECORDS 16.1 Store chromatograms in the study folder. Each chromatogram must have the following information included either in the header or hand unitten on the chromatogram: study number, sample name, extraction date, and dilution factor (if applicable). 16.2 Plot calibration curve by linear regression and ston: in the study folder. 16.3 Print sample list fiom MassLynx and tape into the instrument runlog. 16.4 Print data integration summary fiom MassLynx and tape into the instrument runlog. 16.5 Copy instrument runlog pages, including instrument parameters and sample results, and tape into appropriate study notebook. 16.6 Summarize data using suitable software and store in the study folder. 16.7 Back up electronic data to appropriate medium. Record in study notebook the file name and location of backup electronic data. 17.0 TABLESD,IAGRAMSF,LOWCHARTANSD. VALIDATION DATA 17.1 Attachment A: FACT-M-4 Data reporting spreadsheet 17.2 The validation report associated With this method is FACT-M3.O & 4.0-V-1. 18.0 REFERENCES 18.1 AMDT-EP-3 1 "Operation of VG Platform Electrospray Mass Spectrometer" 19.0 AFFECTEDDOCUMENTS 19.1 FACT-M-3.0, "Extraction of Fluorochemicd Anions from Senun for Analysis Using HPLC-ElectrosprayMasss Spectrometry" 3M Environmental Laboratory FACT-M4.0 Analysis of Serum Extract Using ESMS Page 6 of 8 Page 163 3M Medical Department Study: T6316.1 I Analytical Report: FACT-TOX-001 LRN-U2103 20.0 F&ViSIONS Revision Number. Reason For Revision Revision - Date 3M Environmental Laboratory FACT-M-4.0 Analysis of Serum Extract LJsingESMS Page 7 of 8 Page 164 3M .Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Laboratory Study # Study: Test Material: Matriflinal Solvent: MethodlRevision: Analytical Equipment System Number: Instrument SoftwareNersion: Filename: R-Squared Value: Slope: Y Intercept: Date of Extraction/Analyst: Date of AnalysidAnalyst: Group/Dose: Taken from the study folder. Sample#: Taken from the study folder. Concentration (ug/rnL): Taken from the MassLynx integration summary. Initial Volume (rnL): Taken fiom the study folder. Dilution Factor: Taken from the study folder. Final Cone (ug/rnL): Calculated by dividing the initial volume from the concentration 3M Environmental Laboratory FACT-M-4.0 Analysis of Serum Extract Using ESMS Page 8 of 8 Page 165 3M Medical Department Study: T6316.1 c Analytical Report: FACT-TOX-001 LRN-U2103 3M ENVIRONMENTLAALBORATORY METHOD ANALYSIS OF POTASSIUM PERFLUOR0OC"ESULFONATE OR OTHER FLUOROCHEMICAINLSSERUM[EXTRACTUSING HPLC-ELECTROSPRAY/MAS!~ SPECTROMETRY Method Number: ETS-8-5.1 Author: Lisa Clemen, Robert Wynne Approved By: Adoption Date: 03/01/99 Revision Date: ~ Laboratory Manager ~~ Date Group Leader Date $flc Technical Reviewer .o SCOPE AND APPLICATION 09/2 h i 7 1 Date .l Scope: This method describes the analysis of s e m i extracts for fluorochemical surfactants vD using HPLC-electrospray/mass spectrometry. 21.2 Applicable Compounds:Fluorochernical surfactants or other fluorinated compounds, or other ionizable compounds. 0 2.1.3 Matrices: Rabbit, rat, bovine, monkey, and human serum, or other fluids as designated in '2. the validation report. 3 P, Word 6/95 ETS-8-5.1 Page 1 of 9 3M Environmental Laboratory Analysis of Serum Extract Using ESfMS Page 166 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 2.0 SUMMARY OF METHOD 2.1 This method describes the analysis of fluorochemical surfactants extracted from serum or other fluids, using HPLC-electrospray/massspectrometry, or similar system as appropriate. The analysis is performed by monitoring a single ion characteristic of a particular fluorochemical, such as the perfluorooctanesulfonate (PFOS) anion, m/z= 499. Additionally, samples may be analyzed using a taridem mass spectrometer to further verify the identity of a compound by detecting daughter ions of the parent ion. 3.0 DEFINITIONS 3.1 Atmospheric Pressure Ionization (API): The Micromass Quattro I1 triple quadrupole systems allow for various methods of ionization by utilizing various sources, probes, and interfaces. These include but are not limited to: Electrospray Ionization (ESI), Atmospheric Pressure chemical Ionization (APcI), Thermospray, etc. The ionization process in these techniques occurs at atmosphericpressure (i.e., not under a vacuum). 3.2 Electrospray Ionization (ES,ESI): a method of ionizationperformed at atmospheric pressure, whereby ions in solution are transferred to the gas phase via tiny charged droplets. These charged droplets are produced by the applicatj.onof a strong,electricalfield. 3.3 Mass Spectrometry, Mass Spectrometer (MS), Tandem Mass Spectrometer ( M S / M S ) : The API Quattro 11triple quadrupole systems are equipped with quadrupolemass selective detectors. Ions are selectively discriminatedby mass to charge ratio ( d z ) and subsequently detected. A single MS may be employed for ion detection or a series ( M S M S ) for more specific fi-agmentationinformation. 3.4 Conventional vs. Z-spray probe interface: The latest models of Micromass Quattro II triple quadrupole systems (post 1998) utilize a "Z-spray" conformation. The spray emitted from a probe is orthogonalto the cone aperture. In the conventionalconformationit is aimed directly at the cone aperture, after passing through a tortuous pathway in the counter . electrode. Though the configurationis different,the methods of operation, cleaning, and maintenance are the same. However, Z-spray components and conventional components are not compatiblewith one another, but only with similar systems (Le., Z-spray components are compatiblewith some other Z-spray systems, etc.) 3.5 Mass Lynx Software: System software designed fclr the specific operation of these Quattro I1triple quadrupole systems. CurrentlyMassLynx has Windows 95 and WindowsNT 4.0 versions. All versions are similar. For more details see the manual specific to the instrument (Micromass Quattro I1 triple quadrupole MassLynx or MassLynx NT User's Guide). 4.0 WARNINGS AND CAUTIONS 4.1 Health and Safety Warnings: 4.1.1 Use caution with the voltage cables for the probe. When engaged, the probe employs a voltage of approximately5000 'Volts. 4.1.2 When handling samples or solventswear appropriateprotective gloves, eyewear, and clothing. 3M Environmental Laboratory ETS-8-5.1 Analysis of Serum Extract Using ESMS Page 2 of 9 Page 167 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 4.2 Cautions: 4.2.1 Do not operate solvent pumps above capacity of 400 bar (5800 psi) back pressure. If the back pressure exceeds 400 bar, the HP1100 will initiate automatic shutdown, 4.2.2 Do not run solvent pumps to dryness. 5.0 INTERFERENCES 5.1 To minimize interferenceswhen analyzing samplles, teflon should not be used for sample storage or any part of instrumentationthat comes in contact with the sample or extract. 6.0 EQUIPMENT 6.1 Equipment listed below may be modified in order.to optimize the system. Document any modifications in the raw data as method deviations. 6.1.1 Micromass Quattro 11triple quadrupole Mass Spectrometerequipped with an electrospray ionization source 6.1.2 HP 1100 low pulse solvent pumping system, solvent degasser, column compartment, and autosampler 7.0 SUPPLIES AND MATERIALS 7.1 Supplies 7.1.1 High purity grade nitrogen gas regulated tlo approximately 100psi (House air system) 7.1.2 HPLC analytical column, specificsto be determined by the analyst and documented in the raw data. 7.1.3 Capped autovials or capped 15 mL centrifuge tubes 8.0 REAGENTS AND STANDARDS 8.1 Reagents 8.1.1 Methanol, HPLC grade or equivalent 8.1.2 Milli-QTMwater, all water used in this method should be Milli-QWwater or equivalent, and may be provided by a Mil1.i-QTOC Plus system or other vendor 8.1.3 Ammonium acetate, reagent grade or equivalent 8.2 Standards 8.2.1 Typically two method blanks, two matrix blanks, and eighteen matrix standards are prepared during the extraction procedure. See ETS-8-4.1. 9.0 SAMPLEHANDLING 9.1 Fresh matrix standards are prepared with each analysis. Extracted standards and samples are stored in capped autovials or capped 15 mL cmtrifuge tubes until analysis. 3M Environmental Laboratory ETS-8-5.1 Analysis of Serum Extract Using ES/MS Page 3 of 9 Page 168 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 9.2 If analysis will be delayed, extracted standards and samples can be refrigerated at . approximately 4" C, or at room temperature, until analysis can be performed. 10.0 QUALITY CONTROL 10.1 Solvent Blanks, Method Blanks and Matrix Blranks 10.1.1 Solvent blanks, method blanks and matrix blanks are prepared and analyzed with each batch to determine contamination or carryover. 10.1.2 Analyze a method blank and a matrix blank prior to each calibration curve. 10.2 Matrix Spikes 10.2.1 Matrix spikes are prepared and analyzed to determine the matrix effect on the recovery efficiency. 10.2.2 Matrix spike duplicates are prepared and analyzed to measure the precision and the recovery for each analyte. 10.2.3 Analyze a matrix spike and matrix spike duplicate per forty samples, with a minimum of 2 spikes per batch. 10.2.4 Matrix spike and matrix spike duplicate concentrationswill fall in the mid-range of the initial calibration curve. Additional spike concentrationsmay fall in the lowrange of the initial calibration curve. 10.3 Continuing Calibration Verifications 10.3.1 Continuing calibration verifications are analyzed to veri@ the continued accuracy of the calibration curve. 10.3.2 Analyze a mid-range calibration standard after every tenth sample, with a minimum of one per batch. 11.0 CALIBRATION AND STANDARDIZATION 11.1 Analyze the extracted matrix standards prior to and following each set of extracts. The average of two standard curves will be plotted by linear regression (y = my + b), weighted l/x, not forced through zero, using MassLynx or other suitable software. 11.2 If the curve does not meet requirements, perform routine maintenance or reextract the standard curve (if necessary) and reanalyze. 11.3 For purposes of accuracy when quantitating low levels of analyte, it may be necessary to use the low end of the calibration curve rather than the full range of the standard curve. Example: when attempting to quantitate approximately 10 ppb of analyte, generate a calibration curve consisting of the standards from 5 ppb to 100 ppb rather than the full range of the curve (5 ppb to 1000ppb). This will reduce inaccuracy attributed to linear regression weighting of high concentrationstandards. 3M Environmental Laboratory ETS-8-5.1 Analysis of Serum Extract Using E S N S Page 4 of 9 Page 169 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 12.0 PROCEDURES 12.1 Acquisition Set up 12.1.1 Click on start button in the Acquisition Control Panel. Set up a sample list, Assign a filename using MO-DAY-last digit of year-sample number, assign a method (MS) for acquiring, and type in sample descriptions. 12.1.2 To create a method click on scan button in the Acquisition control panel and select SIR (Single Ion Recording) or MRM. Set Ionization Mode as appropriate and mass to 499 or other appropriatemasses. A full scan is usually collected along with the SIRS. Save acquisition method. If MSMS instruments are employed, additional product ion fiagmentation information ma.1 be collected. See Micromass MassLynx GUIDE TO DATA ACQUISITION for additional information and MRM (Multiple Reaction Monitoring). Time 0.00 min. 8.50 min. 11.Omin. 12.0 min. Ammonium acetate 60% 90% 10% 90% 10% 40% 60% 12.2.2.5 Press the "Start" button. 12.3 Instrument Set-up 12.3.1 Refer to ETS-9-24.0 for more details. 12.3.2 Check the solvent level in reservoirs and refill if necessary. 3M Environmental Laboratory ETS-8-5.1 Analysis of Serum Extract Using ES/MS Page 5 of 9 Page 170 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 12.3.3 Check the stainless steel capillary at the end of the probe. Use an eyepiece to check the tip. The tip should be flat with no jagged edges. If the tip is found to be unsatisfactory, disassemble the probe and replace the stainless steel capillary. 12.3.4 Set HPLC pump to "On". Set the flowto :LO - 500 uL/min or as appropriate. Observe droplets coming out of the tip of the probe. Allow to equilibrate for approximately 10 minutes. 12.3.5 Turn on the nitrogen. A fine mist should he expelled with no nitrogen leaking around the tip of the probe. Readjust the tip of the probe if no mist is observed. 12.3.6 The instrument uses these parameters at thie following settings. These settings may change in order to optimize the response: 12.3.6.1 Drying gas 250-400 litershour 12.3.6.2 ESI nebulizing gas 10-15 litershur 12.3.6.3 HPLC constant flow mode, flow rate 10- 500 pL/min 12.3.6.4 Pressure <400 bar (This parameter is not set, it is a guide to ensure the HPLC is operating correctly.) 12.3.7 Carefully guide the probe into the opening. Insert probe until it will not go any further. Connect the voltage cables to the probe. 12.3.8 Print the tune page, with its parameters, arid store it in the study binder with a copy taped into the instrument log. 12.3.9 Using the cross-flow counter electrodein the ESMS source is recommended for the analysis of biological matrices. 12.3.10Click on start button in the Acquisition Control Panel (this may vary among MassLynx versions, see appropriate MasslLynx USER`S GUIDE). Press the start button. Ensure start and end sample number includes all samples to be analyzed. 13.0 DATAANALYSIASND CALCULATIONS 13.1 Calculations: 13.1.4 Calculate matrix spike percent recoveries using the following equation: % Recovery = Observed Result - Backaound Result x 100 Expected Result 13.1.5 Calculate percent difference using the following equation: % Difference = ExDected Conc. - Calculated Conc. x 100 Expected Conc. 13.1.6 Calculate actual concentration of PFOS, or other fluorochemical,in matrix (pg/mL): Inn of PFOS calc. from std. Curve x Dilution Factor) x 1 c1q (Initial Volume of matrix (mL) + mL of Surrogate Standard) 1000 ng Final Volume (mL) 3M Environmental Laboratory ETS-8-5.1 Analysis of Serum Extract Using ESMS Page 6 of 9 Page 171 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 14.0 METHODPERFORMANCE 14.1 Method Detection Limit (MDL) and Limit of Quantitation (LOQ) are method, analyte, and matrix specific. Please see ETS-8-4.1, Attachment B, for a listing of current validated MDL and LOQ values. 14.2 Solvent Blanks, Method Blanks, and Matrix Bkanks 14.2.1 Solvent blanks, method blanks, and matrix blanks values are must be below the lowest standard in the calibration curve 14.3 Calibration Curves 14.3.1 The I? value for the calibration curve must be 0.980 or better. 14.4 Matrix Spikes 14.4.1 Matrix spike percent recoveries are must be within k 30% of the spiked concentration. 14.5 Continuing Calibration Verifications 14.5.1 Continuing calibration verification percent recoveries must be k 30% of the spiked concentration. 14.6 If criteria listed in this method performance section isn't met, maintenance may be performed on the system and samples reanalyzed or other actions as determined by the analyst. Document all actions in the appropriate logbook. 14.7 If data are to be reported when performance criteria have not been met, the data must be footnoted on tables and discussed in the text of the report. 15.0 POLLUTION PREVENTION AND WASTE MANAGEMENT 15.1 Sample extract waste and flammable solvent is disposed in high BTU containers, and glass pipette waste is disposed in broken glass containers located in the laboratory. 16.0 RECORDS 16.1 Each page generated for a study must have the following information included either in the header or hand written on the page: study or project number, acquisitionmethod, integrationmethod, sample name, extraction date, dilution factor (if applicable), and analyst. 16.2 Print the tune page, sample list, and acquisition method from MassLynx to include in the appropriate study folder. Copy these pages and tape into the instrument runlog. 16.3 Plot the calibration curve by linear regression, weighted l/x, ihen print these graphs and store in the study folder. 16.4 Print data integration summary, integration method, and chromatograms, from MassLynx, and store in the study folder. 3M Environmental Laboratory ETS-8-5.1 Analysis of Serum Extract IJsing ESMS Page 7 of 9 Page 172 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 16.5 Summarize data using suitable software (Excel 5.0) and store in the study folder, see Attachment A for an example of a summary spreadsheet. 16.6 Back up electronic data to appropriate medium. Record in study notebook the file name and location of backup electronic data. 17.0 TABLESD,IAGRAMSF,LOWCHARTASN,D VALIDATION DATA 17.1 Attachment A: ETS-8-5.1 Data summary spreadsheet. 18.0 REFERENCES 18.1 FACT-M-4.1,"Extraction of Potassium Perfluoroloctanesulfonateor Other Fluorochemical compounds from Serum for Analysis Using HPLC-ElectrospraylhlassSpectrometry 18.2 ETS-9-24.0, "Operation and Maintenance of the Micromass Atmospheric Pressure Ionization/Mass Spectrometer Quattro I1 triple q~iidrupoleSystems" 18.3 The validation report associated with this method is ETS-8-4.0 & 5.0-V-1. 19.0 AFFECTEDOCUMENTS 19.1 ETS-8-4.1,"Extraction of Potassium Perfluorooctatnesulfonate or Other Fluorochemical Compounds from Serum for Analysis Using HPLC-ElectrosprayMass Spectrometry" 20.0 REVISIONS Revision Number. 1 Reason For Revision Section 6.1.2 Clarification of HP1100 system components. Section 11.1 Average of two curves, not standard values, are used for plotting linear regression and added the l/x weighting of the curve. Section 12.2.2.4 Clarification of solvent ramp. Section 17.1 Changed from attachment B to A. Revision 04/02/99 3M Environmental Laboratory ETS-8-5.1 Analysis of Serum Extract Uzing ESMS Page 8 of 9 Page 173 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Laboratory Study # Study: Test Material: Matriflinal Solvent: MethodRevision: Analytical EquipmentSystemNumber: Instrument SoftwareNersion: Filename: R-Squared Value: Slope: Y Intercept: Date of ExtractiodAnalyst: Date of Analysis/Analyst: Group Dose Sample# Concentration ug/d Initial Vol. mL Dilution Factor Final Conc. ug/mL Slope: Taken from linear regression equation. Attachment A: Summary Spreadsheet ETS-8-5.1 3M Environmental Laboratory Analysis of Serum Extract Using ESMS Page 9 of 9 Page 174 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 3M ENVIRONMENTLAALBORATORY METHOD ANALYSIS OF POTASSIUM PERF'LUOROOCTANESU14FONATEOR OTHER FLUOROCHEMICINALLISVER EXTRACTUSSING HPLC-ELECTROSPRAYMSAPSESCTROMETRY Method Number: ETS-8-7.0 Author: Lisa Clemen, Glenn Langenburg Adoption Date: 0?/2.zl T f Revision Date: Nfi Approved By: Group Leader C\-PPNR LA. Technical Reviewer I Date 3 / 14/33 Date Date I 3oL J 1.1 Scope: This method is for the analysis of mLC-electrospray/mass spectrometry. liver extracts for fluorochemical surfactants using Applicable Compounds:Fluorochemical surfactants or other fluorinated compounds, or other ionizable compounds. Matrices: Rabbit, rat, bovine, monkey liver, or other tissues as designated in the validation report. Word 6/95 3M Environmental Laboratory ETS-8-7.O Analysis of Liver Extract Using ES/MS Page 1 of 10 Page 175 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 2.0 SUMMAROYF METHOD 2.1 This method describes the analysis of fluorochemicalsurfactants extracted from liver using HPLC-electrospray/massspectrometry, or similar system as appropriate. The analysis is performed by monitoring a single ion characteristic of a particular fluorochemical, such as the perfluorooctanesulfonate(PFOS) anion, m/z =: 499. Additionally, samples may be analyzed using a tandem mass spectrometer to further verify the identity of a compound by detecting daughter ions of the selected parent ion. 3.0 DEFINITIONS 3.1 Atmospheric Pressure Ionization (MI): The Micromass Quattro IItriple quadrupole systems allow for various methods of ionization by utilizing various sources, probes, and interfaces. These include but are not limited to: Electrospray Ionization (ESI), Atmospheric Pressure chemical Ionization (APcI), Thermospray, etc. The ionization process in these techniques occurs at atmosphericpressure (Le. not under a vacuum). 3.2 Electrospray Ionization (ES, ESI): a method of ioilization performed at atmospheric pressure, whereby ions in solution are transferred to the gas phase via tiny charged droplets. These charged droplets are produced by the application of a strong electrical field. 3.3 Mass Spectrometry,Mass Spectrometer (MS), Tandem Mass Spectrometer (MSMS): The MI Quattro I1triple quadrupolemass spectrometer is equipped with two quadrupole mass selective detectors and a collision cell. Ions are selectively discriminated by mass to charge ratio ( d z ) and subsequentlydetected. A single MS may be employed for ion detection or an ion may be selected in the first quadrupole, fragmented in the collision cell, and these fragments may be analyzed in the second quadrupole. 3.4 Conventional vs. Z-spray probe interface: The latest models of Micromass Quattro I1 triple quadrupole (post 1998)utilize a "Z-spray" conformation. The spray emitted from a probe is orthogonal to the cone aperture. In the conventionalconformationit is aimed directly at the cone aperture, after passing through a tortuous pathway in the counter electrode. Though the configuration is different, the methods of operation, cleaning, and maintenance are the same. However, Z-spray Components and conventional components are not compatible with one another, but only with similar systems (Le. Z-spray components are compatible with other Z-spray systems, etc.) 3.5 Mass Lynx Software: System software designed for the specific operation of these Quattro 11triple quadrupole systems. Currently MassLynx has Windows 95 and WindowsNT 4.0 versions. All versions are similar. For more details refer to the manual specific to the instrument (Micromass Quattro I1 triple quadrupole MassLynx or MassLynx NT User's Guide). 4.0 WARNINGS AND CAUTIONS 4.1 Health and Safety Warnings: 4.1.1 Use caution with the voltage cables for the probe. When engaged, the probe employs a voltage of approximately 5000 `Volts. 3M Environmental Laboratory ETS-8-7.0 Analysis of Liver Extract Using ESMS Page 2 of 10 Page 176 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 4.1.2 When handling samples or solvents wear appropriate protective gloves, eyewear, and clothing. 4.2 Cautions: 4.2.1 Operate the solvent pumps below a back pressure of 400 bar (5800 psi). If the back pressure exceeds 400 bar, the HPllOO will initiate automatic shutdown. 4.2.2 Do not run solvent pumps to dryness. 5.0 INTERFERENCES 5.1 To minimize interferences when analyzing samp1cs, Teflon shall not be used for sample storage or any part of instrumentationthat comes in contact with the sample or extract. 6.0 EOUIPMENT 6.1 Equipment listed below may be modified in order to optimize the system. Document any modifications in the raw data as method deviations. 6.1.1 Micromass Quattro II triple quadrupole Mass Spectrometer equipped with an electrospray ionization source. 6.1.2 HP1100 low pulse solvent pumping systern, solvent degasser, column compartment, and autosampler 7.0 SUPPLIES AND MATERIALS 7.1 Supplies 7.1.1 High purity grade air regulated to approximately 100psi (house air system) 7.1.2 HPLC analytical column, specifics to be determined by the analyst and documented in the raw data 7.1.3 Capped autovials or capped 15 ml centrifuge tubes 8.0 REAGENTASND STANDARDS 8.1 Reagents 8.1.1 Methanol, HPLC grade or equivalent 8.1.2 Milli-QTMwater (ASTM type I), all water used in this method should be ATSM type I, or equivalent, and be provided by it Milli-Q TOC Plus system or other vendor 8.1.3 Ammonium acetate, reagent grade or equivalent 8.1.3.1 When preparing different amounts than those listed, adjust accordingly. 8.1.3.2 2.0 mM ammonium acetate solutjon: Weigh approximately 0.300 g ammonium acetate. Pour into a 2000 mL volumetric container containing 2000 mL Milli-QTMwater, mix until all solids are dissolved. Store at room temperature. 3M Environmental Laboratory ETS-8-7.0 Analysis of Liver Extract Using E S N S Page 3 of 10 Page 177 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 8.2 Standards 8.2.1 Typically two method blanks, two matrix blanks, and eighteen matrix standards are prepared during the extraction procedure. Refer to ETS-8-6.0. 9.0 SAMPLHEANDLING 9.1 Fresh matrix standards are prepared with each analysis. Extracted standards and samples are stored in capped autovials or capped 15 ml centrifuge tubes until analysis. 9.2 If analysis will be delayed, extracted standards arid samples may be stored at room temperature, or refiigerated at approximately4' C, until analysis can be performed. 10.0 OUALITYCONTROL 10.1 Method Blanks and Matrix Blanks 10.1.1 Solvent blanks, method blanks, and matrix blanks are prepared and analyzed with each batch to determine contamination or carryover. 10.1.2 Analyze a method blank and a matrix blank prior to each calibration curve. 10.2 Matrix Spikes 10.2.1 Matrix spikes are prepared and analyzed to determine the matrix effect on the recovery efficiency. 10.2.2 Matrix spike duplicates are prepared and analyzed to measure the precision and the recovery for each analyte. 10.2.3 Analyze a matrix spike and matrix spike d!uplicateper forty samplep. With a minimum of 2 spikes per batch. 10.2.4 Matrix spike and matrix spike duplicate concentrationswill fall in the mid-range of the initial calibration curve. Additional spike concentrations may fall in the low- range of the initial calibration curve. 10.3 Continuing Calibration Checks 10.3.1 Continuing calibration verifications are analyzed to veri@ the continued accuracy of the calibration curve. 10.3.2 Analyze a mid-range calibration standard every tenth sample, with a minimum of one per batch. 11.0 CALIBRATION AND STANDARDIZATION 11.1 Analyze the extracted matrix standards prior to arid following each set of sample extracts. The average of two standard curves will be plotted by linear regression (y = mx -t- b), weighted l/x, not forced through the origin, using MassLynx or other suitable software. 11.2 If the curve does not meet requirements perform routine maintenance or reextract the standard curve (if necessary) and reanalyze. 3M Environmental Laboratory ETS-8-7.0 Analysis of Liver Extract Using ES/MS Page 4 of 10 Page 178 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 11.3 For purposes of accuracy when quantitating low kvels of analyte, it may be necessary to use the low end of the calibration curve rather than the full range of the standard curve. Example: when attempting to quantitate approxirnately 10 ppb of analyte, generate a calibration curve consisting of the standards from 5 ppb to 100ppb rather than the full range of the curve (5 ppb to 1000ppb). This will reduce inaccuracy attributedto linear regression weighting of high concentration standards. 12.0 PROCEDURES 12.1 Acquisition Set up 12.1.1 Set up the sample list. 12.1.1.1 Assign a sample list filename using MO-DAY-last digit of year-increasing letter of the alphabet starting with a 12.1.1.2 Assign a method ( M S file) for acquiring 12.1.1.3 Assign an HPLC program (Inlet iile) 12.1.1.4 Type in sample descriptions and vial position numbers 12.1.2 To create a method click on method in the Acquisition control panel then mass spectrometer headings and select SIR (Single Ion Recording) or MRM (Multiple Reaction Monitoring). Set Ionization Mode as appropriate and mass to 499 or other appropriate masses. A full scan is usually collected along with the SIRS. Save acquisition method. If MSMS instruments are employed, additional product ion fragmentation information may be collected. Refer to Micromass MassLynx GUIDE TO DATA ACQUISITION for additional information and MFW. 12.1.3 Typically the analytical batch run sequencle begins and ends with a set of extracted matrix standards. 12.1.4 Samples are analyzed with a continuing calibration verification injected standard after every tenth sample. Solvent blanks should be analyzed periodically to monitor possible analyte carryover and are not considered samples but may be included as such. 12.2 Using the Autosampler 12.2.1 Set up sample tray according to the sample: list prepared in Section 12.1.1. 12.2.2 Set-up the HPllOO/autosampler at the following conditions or at conditions the analyst considers appropriate for optimal response. Record actual conditions in the instrument logbook: 12.2.2.1 Sample size = 10 pL injection 12.2.2.2 InjecVsample = 1 12.2.2.3 Cycle time = 9 minutes 3M Environmental Laboratory ETS-8-7.0 Analysis of Liver Extract Using ESMS Page 5 of 10 Page 179 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 12.2.2.4 Solvent ramp conditions I 0.00min. I 40% 1.0 min. I 40% I 4.5min. I 95% 7.0 min. 40% 9.0 mi. 40% I Ammoniumacetate I 1 60Yn 1 1 I 60% 1 I I 5% 60% 12.2.2.5 Press the "Start" button. 12.3 Instrument Set-up 12.3.1 Refer to ETS-9-24.0, "Operation and Maintenance of the Micromass Quattro I1 Triple Quadrupole Mass SpectrometerFitted with an Atmospheric Pressure Ionization Source," for more details. 12.3.2 Check the solvent level in reservoirs and rafill if necessary. 12.3.3 Check the stainless steel capillary at the end of the probe. Use an eyepiece to check the tip. The tip should be flat with no jagged edges. If the tip is found to be unsatisfactory, disassemble the probe and replace the stainless steel capillary. 12.3.4 Turn on the nitrogen. 12.3.5 Open the tune page. Clicks on operate to initiate source block and desolvation heaters. 12.3.6 Open the Inlet Editor. 12.3.6.1 Set HPLC pump to "On" 12.3.6.2 Set the flow to 10 - 500 uL/min or as appropriate 12.3.6.3 Observe droplets coming out of the tip of the probe. A fine mist should be expelled with no nitrogen leaking around the tip of the probe. Readjust the tip of the probe if no mist is o`bserved 12.3.6.4 Allow to equilibrate for approximately 10 minutes. 12.3.7 The instrument uses these parameters at the following settings. These settings may change in order to optimize the response: 12.3.7.1 Drying gas 250-400 litershour 12.3.7.2 ESI nebulizing gas 10-15 litershour 12.3.7.3 HPLC constant flow mode flow rate 10- 500 pL/min 12.3.7.4 Pressure <400 bar (This parameter is not set, it is a guide to ensure the HPLC is operating correctly.) 12.3.7.5 Source block temperature 150' 12.3.7.6 Desolvation temperature 250" 3M Environmental Laboratory ETS-8-7 .O Analysis of Liver Extract Using ESMS Page 6 of 10 Page 180 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 12.3.8 Print the tune page, with its parameters, and store it in the study binder with a copy taped into the instrument log. 12.3.9 Click on start button in the Acquisition Control Panel (this may vary among MassLynx versions, refer to appropriate RlassLynx User's Guide). Ensure start and end sample number includes all samples to be analyzed. 13.0 DATAANALYSIASND CALCULATIONS 13.1 Calculations: 13.1.4 Calculate matrix spike percent recoveries using the following equation: % Recovery = Observed Result - Backmourtd Result x 100 Expected Result 13.1.5 Calculate percent difference using the following equation: % Difference = Expected Conc. - Calculated Conc. x 100 Expected Conc. 13.1.6 Calculate actual concentrations in matrix (pg/g): (ncz of PFOS calc. from std. Curve x Dilution Factor) (Initial Weight of'Liver (a) Final Volume (mL) x 1 up, 1000 ng 14.0 METHODPERFORMANCE 14.1 Method Detection Limit (MDL) and Limit of Quatntitation (LOQ) are method, analyte, and matrix specific. Refer to ETS-8-6.0, Attachment B for a listing of current validated MDL and LOQ values. . 14.2 Solvent Blanks, Method Blanks and Matrix Blanks 14.2.1 Solvent blanks, method blanks, and matrix blanks must be below the lowest standard in the calibration curve. 14.3 Calibration Curves 14.3.1 The 3 value for the calibration must be 0.980 or better. 14.4 Matrix Spikes 14.4.1 Matrix spike percent recoveries must be within rt 30% of the spiked concentration. 14.5 Continuing Calibration Verification 14.5.1 Continuing calibration verificationpercent recoveries must be within 1- 30% of the spiked concentration. 14.6 If criteria listed in the method performance section are not met, maintenance may be performed on the system and samples reanalyzed or other actions as determined by the analyst. Document all actions in the appropriate logbook. 3M Environmental Laboratory ETS-8-7.0 Analysis of Liver Extract U:;ing ES/MS Page 7 of 10 Page 181 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 14.7 If data are to be reported when performance criteria have not been met, the data must be footnoted on tables and discussed in the text of the report. 15.0 POLLUTION PREVENTION AND WASTE MANAGEMENT 15.1 Sample extract waste and flammable solvent is disposed in high BTU containers, and glass pipette waste is disposed in broken glass containers located in the laboratory. 16.0 RECORDS 16.1 Each page generated for a study must have the following information included either in the header or hand Written on the page: study or project number, acquisition method, integration method, sample name, extraction date, dilution factor (if applicable), and analyst. 16.2 Print the tune page, sample list, and acquisition method from MassLynx to include in the appropriate study folder. Copy these pages and tape into the instrument runlog. 16.3 Plot the calibration curve by linear regression, weighted l/x, then print these graphs and store in the study folder. 16.4 Print data integration summary, integration method, and chromatograms from MassLynx and store in the study folder. 16.5 Summarize data using suitable s o h a r e (Excel 5.0+) and store in the study folder, refer to Attachment A for an example of a summary spreadsheet. 16.6 Back up electronic data to appropriatemedium. Fkecord in study notebook the file name and location of backup electronic data. 17.0 TABLESD, IAGRAMFS,LOWCHARATNSD, VALIDATION DATA 17.1 Attachment A: ETS-8-7.0 Data summary spreadsheet 18.0.REFERENCES 18.1 FACT-M-2.1, "Extraction of Potassium Perfluorooctanesulfonateor Other Fluorochemical Compounds from Liver for Analysis Using HPLC!-Electrospr;iy/Mass Spectrometry" 18.2 ETS-9-24.0, "Operation and Maintenance of the Micromass Atmospheric Pressure IonizationMass SpectrometerQuattro I1triple quadrupole Systems" 18.3 The validation report associated with this method is ETS-8-6.0 & 7.0-V-1 19.0 AFFECTED DOCUMENTS 19.1 ETS-8-6.0, "Extraction of Potassium Perfluorooctanesulfonateor Other Fluorochemical Compounds from Liver or Fluid for Analysis Using HPLC-Electrosprayblass Spectrometry" 3M Environmental Laboratory ETS-8-7.0 Analysis of Liver Extract U:,ing ES/MS Page 8 of 10 Page 182 3M .Medical Department Study: T6316.1 20.0 REVISIONS Revision Number Analytical Report: FACT-TOX-001 LRN-U2103 Reason For Revi!;ion Revision 3M Environmental Laboratory ETS-8-7.0 Analysis of Liver Extract Using ESMS Page 9 of 10 Page 183 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Laboratory Study # Study: Test Material: Matrix/Final Solvent: MethodIRevisi on: Analytical Equipment System Number: Instrument SoftwareNersion: Filename: R-Squared Value: Slope: Y Intercept: Date of ExtractiodAnalyst: ' Date of AnalysislAnalyst: Group Dose Sample# Concentration Wg lnitiai wt. g Uilution Factor Final Conc. wk Attachment A: Sunlmary Spreadsheet ETS-8-7.0 3M Environmental Laboratory Analysis of Liver Extract Using ES/MS Page 10 of 10 Page 184 3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 Appendix D: Data Summary Tables Analytical Report: FACT-TOX-001 LRN-U2103 Analytical Report: FACT TOX-001 LRN-U2103 3M Environmental Laboratory Page 185 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 .. h d 0 E LL b 3 3 3M Environmental Laboratory Page 186 3M Medical Department Study: T6316.1 1 A Analytical Report: FACT-TOX-001 LRN-U2103 3M Environmental Laboratory -- Page 187 3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 Analytical Report: FACT TOX-001 LRN-U2103 Table 12. FACT TOX-001 Data Summary of PFOSAA Concentration-Serum (pg/mL) Analytical Report: FACT-TOX-001 LRN-U2103 b L W m i tof Quantitation= 0.0248p@mL ' L W m i t of Quantitation= 0.00493p@mL dLOQ--Limit of Quantitatjon= 0.0097 *AnimalC91216Fsamplewas lost duringextraction. NOTE: Results are expressed as grwp/genderaverages *the standaddeviation associatedwith that grcup/gender. NOTE: The only measurementof accuracyawilableatthis time, matrixspike studies, indicatethatthe sera and liver data canbe consideredaccurateto within one standarddeviationof the average fortifiedsamples recovery.The averagefortifiedsamplerecoverywas 109%with a standarddeviationof 23%. 3M Environmental Laboratory Page 188 3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 Analytical Report: FACT TOX-001 LRN-U2103 Analytical Report: FACT-TOX-001 LRN-U2103 Table 13. FACT TOX-001 Data Summary of EtFOSE-OHConcentration-Serum (pg/mL) a Not correctedfor purityof the standardmaterial. bLOQ--Limitof Quantitation= 0.00977pgmL CLOC+Limit of Quantitation= 0.0248 pgmL dLO(tlimit of Quantitation= 0.00493ugmL `Animal C91216F sample was lost during extraction. NOTE: Resultsare expressed as grouplgenderaveragesfthe standard dm'ationassociated with that group'gender. NOTE: The resultsof qualiiycontrolanabes (curvefit, CCVs, and MSiMSDs) for EtFOSE-OHwere inconsistentand indicatethat datapresentedshouldbe consideredto be qualitativeonly. Values are presentedinthis data table inthe spiritof full disclosure,butshouldnotbe usedinanyquantitativeassessmentof the data. 3M Environmental Laboratory Page 189 3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 Analytical Report: FACT TOX-001 LRN-U2103 Table 14. FACT TOX-001 Data Summary of M556 Concentration-Serum (pg/mL) Analytical Report: FACT-TOX-001 LRN-U2103 Male I I I I I I Week 14a week 2 7 Male Female I Male (<nL=O1C0Ib) II 1I Week53" Male Female Male <LOCIC (n = 15) <LOQC (n=15) In = 14) Female <LOQC (n = 13) 0.680f 0.667 (n = 11)" 3.02 i 1.08 (n = 15) 5.06 f 1.39 (n = 19) <LO@ (n = 12) a Not correctedfor puntyof the standardmaterial. bLOQ--Limitof Quantitation= 0.0248 p g h L 'LOQ-Limit of Quantitation= 0.00494pglrnL 'Animal C91216F sample lost during extraction. NOTE: Resultsare eqxessed as group/gender averages*the standarddeviation associatedv4th that group/gender. NOTE: The miy measurementof accuracyavailaMe at this time, matrixspike studies. indicatethat the sem and l i i r data can beconsideredaccumteto withinm e standardhiationof the aemge fortified samples m.The a e m ~ f w t i f i e dsample~BCO\RTYwas 123%with a standarddeviationof 20%. 3M Environmental Laboratory Page 190 3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 Analytical Report: FACT TOX-001 LRN-U2103 Table 15. FACT TOX-001 Data Summary of PFOSEA Concentration-Serum , + g h Analytical Report: FACT-TOX-001 LRN-U2103 b L W m i tof Quantitation= 0.0247 pg'mL "LOQ-4imit of Quantitation= 0.W92 pdmL d L W m i tof Quantitation= 0.00975ps/mL ' L O W m i t of Quantitation= 0.04% pg'mL 'Animal C91216F sample lost during exhaction. NOTE: Resub areexpressedasgroupgenderaverages ithestandardd a t i o n associatedwith that group/gendet. NOTE: The resultsof qualitycontrolanalyses(CUMfit, CCVs, and IVSMSDs) for PFOSEAwere inconsistentand indicatethat data presentedfor this anatyte shouldbe consideredto be qualitatie only. Values are presentedhereinthe spirit of full disclosure,but shouldnotbe usedinanyquantitativeassessmentof the data. 3M Environmental Laboratory Page 191 3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 Analytical Report: FACT TOX-001 LRN-U2103 Table 16. FACT TOX-001 Data Summary of PFOS Concentration-Liver (pg/g) Analytical Report: FACT-TOX-001 LRN-U2103 Male Week 8" 2.13i0.668 (n = 5) 1067 f 197 (n = 5) Week 27 I Week53 Male Male I 0.571 f 0.175 (n = 5) Week 105 ivide Female I 0.143io.133 I ?.56*7.!c! I 90.j *?C.3 1 (n = 18) (n = 22) (n = 22) I I I 0.178f 0.111 38.5f 21.9 127 i 68.5 (n = 14) (n = 13) (n = 15) 695 f 189 (n = 5) 313 i 22G I (n = 17) 297 f 171 (n = 19) I 2.05 f 3.36 (n = 10) 7.69f 7.62 (n = 12) 3M Environmental Laboratory Page 192 3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 Analytical Report: FACT TOX-001 LRN-U2103 Table 17. FACT TOX-001 Data Summary of PFOSA Concentration-Liver (pg/g) Timepoint Sex Group 1 Control Average &D Group 2 Low Average t S D Group 3 Mid Average &D Group 4 Mid-High Average &D Analytical Report: FACT-TOX-001 LRN-U2103 Group 5 High Average &D Group 6 Mid-High Recovery Average SD Week 8" Male 0.0313 i 0.00163 (n = 5) 0.0310i 0.00189 19.2 f 0.987' (n = 5) I Female (n = 5) I 1 I 1.80*0.0831 (n = 5) 7.98k1.03 (n = 5) 14.6 f 1.52 (n = 5) I Male week27 I I I I I I I Week53" I Male Female 0.0182 i u.0100 (n = 5) (n = 5) I 28.2 i 8.44 (n = 5) (n = 5) I bLOQ-Limitof Quantitatim= 0.0123 pg'g 'LOQ-Limit of Quantitation= 0.00614pg'g 'Animal C90748M sample lost during extraction. %CVs failed, data enteredas estimated. NOTE Results are expressedas group/genderaveragesithe standard deviationassociated with that group/gender. NOTE: The only measurementof accuracyavailableatthis time, matrixspike studies,indicatethat the sera andliver datacanbeconsideredaccurateto withinone standarddeviationof the average fortifiedsamples recovery.The averagefortifiedsamplerecoverywas 102%with a standarddeviationof 17%(revisedresults). 3M Environmental Laboratory Page 193 3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 Analytical Report: FACT TOX-001 LRN-U2103 Table 18. FACT TOX-001 Data Summary of PFOSAA Concentration-Liver (pg/g) Analytical Report: FACT-TOX-001 LRN-U2103 C L O W m i tof Quantitation= 0 0123 p$g 'Animal C90748M lost during exlraction. NOTE:Resultsare expressed as grWp/gender averagesfthe standarddeviationassociatedwiththat group/gender. NOTE: The onb measurementof accuracyawilableat this time, rnatnxspke studies, indicatethat the sera and lwrdata can be msidered accurateto wlthinone standarddewation of the awragefottlfied samples m r y The awragefortdied sample remery was 105% wlth a standalddevlatim of 19%(msed resuks) 3M Environmental Laboratory Page 194 3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 Analytical Report: FACT TOX-001 LRN-U2103 Analytical Report: FACT-TOX-001 LRN-U2103 Table 19. FACT TOX-001 Data Summary of EtFOSE-OHConcentration-Liver (pglg) 1 1 1 1 Timepoint Sex Group 1 Control Average tSD Group 2 Low Average *SD Group 3 Mid Average +SD Group 4 Mid-High Average S D II II week4" Male Female ;Lmnb=5) (n = 5) Group 5 High Average A D Group 6 * Mid-High Recovery Average SD bLOQ--Limit of Quantitation = 0.0596 pg'g ' L W m i t of Quantitation= 0.119pg'g dLOQ-Limit of Quantitation = 0.0298 pg'g 'LOQ--Limit of Quantitation= 0.0614 pg'g NA-Not applicable NR-Not reported 'Animal C91216F sample lost during earaction. NOTE: Results areeqxessed as group/gender%rages *the standard deviationassociatedwith that grcup/gender. NOTEThe resultsof qualitycontrol analyses (cumfit, CCVs, and fvWvtSDs)for EtFOSE-OHwere inconsistentand indicatethat data presentedfor this analyteshouldbe consideredto be qualitative only. Values are presentedhereinthe spiritof fulldisclosure, but should not be usedin anyquantitativeassessmentof the data. 3M Environmental Laboratory Page 195 3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 Analytical Report: FACT TOX-001 LRN-U2103 Table 20. FACT TOX-001 Data Summary of M556 Concentration-Liver (pg/g) Analytical Report: FACT-TOX-001 LRN-U2103 Weeksa I week 10sa Male Female Male Female <LOQ~ (n =5) <LOCIb (n = 51 I pt,Sb,e i V . W 2 V.JI 0 I 9.5i f 3 . B I (n = 18) (n=22) (n = 22) <Lab$ (n = 14) 2.03f 2.74 (n = 13) 8.57f 3.33 (n = 15) 58.9f 17.0 (n = 5) 36.3 f 9.51 (n = 5) 27.1 f 9.41 (n = 17) 28.5f 8.86 (n = 19) <LOQC,d (n = 10) <LO@* (n = 12) a Not corrected for purityof the standard material. b L w C i r n i tnf ~uzr?!i~ti=wP! . P ~ cLOQ-Limit of Quantitation= 0.00615ps/s dLOQ-Lirnitof Quantitation= 0.123 pg'g 'LOQ-Lirnit of Quantitation= 0.0123 ps/s ' L W m i t of Quantitation= 0.0615 pg'g NOTE Resultsare expressed as grouplgender ayerages f the standard deviation associatedwiththat group/gender. NOTE: The mlymeasurementof accuracyavailableat his time, matrixspikestudies,indicatethat the sera and lmrdata canbe msideredaccurate to within m e standarddeviationof the awrap fortified samples recwry. The awragefortifiedsample recoverywas 102%with a standardMatim of 15%. 3M Environmental Laboratory Page 196 3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 Analytical Report: FACT TOX-001 LRN-U2103 Analytical Report: FACT-TOX-001 LRN-U2103 Timepoint week4 Week8 Week 14 Group 1 Group 2 Group 3 Group 4 Group 5 Sex Control Low Mid Mid-High High Average &D Average tSD Average tSD Average +SD Average tSD Male I I I I I Group 6 Mid-High Recovery Average t SD Male I I I I I J Male I I I 1 a Not correctedfor purityof the standardmaterial bLOQ--Limitof Quantitation= 0.0307ps/s "LOQ-Limit of Quantitation= 0.0613 ps/s NOTE: Results are expressed as grwp/aenderm m ~ ethe stmdzc!devk??sasswigedvv%iWit giiX@@iid~i. NOTE: The resultsof qualitycontrolanalyses(curvefit,CCVs, andlvWMSDs)for PFOSEAwere inconsistentandindicatethat data presentedforthis analyteshouldbe consideredto be qualitatieonly. Values are presentedhereinthe spiritof full disclosure,butshouldnotbe used inanyquantitatk assessmentof the data. 3M Environmental Laboratory Page 197 3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 Appendix E: Data Spreadsheets Analytical Report: FACT-TOX-001 LRN-U2103 Analytical Report: FACT TOX-001 LRN-U2103 3M Environmental Laboratory Page 198 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance# 6329-212 St"@ product N w n k Q e r t Subslance) Sample Data I'EEK4 RAT SER Group Dose REWORK Snmplr x Group 2 Low Dore 3 0 mgng Group 3 Mid Dose 30 0 mgnig H20 Elk-! H 2 0 Elk-2 Rat Serum Blk-l Rat SenunBlk-2 MS 4/23/98 MSD 4123198 MS 4128198 MSD 4/28/98 C90719M C90732M C90725M C90753M C90775M C91135F CY1 143F C91165F CY1169F C91171F C90789M C90814M C90817M C90833M C90834M C91198F C91205F C91222F C9123OF C91237F C90846M C90853M C90866M /.-"".., C90890M b,"Y,III. C91251F C91272F C91285F C91287F C91291F C9093SM C90939M Group 5 C91333F C9134SF C91350F C90984M CYIWSM CYlOZ4M C91033.M C91037M C91423F C91427F C91430F C91443F C.alld*F 104 Week Dietary Carcinogenicity Sludy with N m w Range (98 1%) N-E?h)lPerfluorwctanerulfonamldoEthanol m Rats T-6316 (EIFOSEOH) Rat Serum FACTM-3 0 & FACT-MM4 0 reuorkrd u m g ETS-8-5 I Filenames Chick 080697 and Madelme 041098 PFOS MassL- 2 3 & 3 I BLS 13042780243 & 59-60 See llsllng 10 Ule "ghl See Attachments See Attachments See Attachments 04123198,041281Y0 RWW 04127198,04128198,04/30198,05105198 KIWHOJ crpl crpz crp3 crp4 Grp 5 R04278 13-24 R05058 12-23 RO505826-37 R05058M-51 R0428813-24 09'01'W, 12101'W, 12/04'W, 12!05'W M M H W H 412311938 MS,MSD R0427856-57 ErtTnctIon Val. mL I I I I I I I I I I I I I I - PFOS Std - COWHtiO" FXIW 0 9275 0 9275 ~ 0 9275 0 9275 ~ NA - - NA NA NA 0 9275 0 9275 0 9275 0 9275 - 0 9275 0 9275 Lot215 PFOSPurity Comerdon Fanor unknown Unknown unknown unknown NA NA NA NA Unknown UnknOUn Unknown unknoxn unknown unknown 412811938 MS, MSD PFOS Dilution Fsrtar 1 1 I 1 I I I I 1 1 I 1 1 1 PFOS Canr. nc,ImL OW 0680 OW 248 240 233 161 457 263 325 263 80 I 534 120 rnk"mn R0427802 R0427859 R0427803 R04278M u.n~427856 R0427857 RO505852 R0505853 R04278 I3 RIM27814 R0427815 R0427816 R0427817 R0427820 Cancmlmthln Of PFOS ugh& or *h RH <LoQ(0 0231 u g i d ) 4LoQ (00231 u g i d ) CLOQ (00231 u g l d ) <LOQ (00231 ug/mL) 96% 93% 65% 183% 0 0244 0 0302 0 0244 0 0743 0 0495 0 111 RO505852-53 MIS" PFOS U@mL CLOQ LoQ 95% 124% 0 OM5 1 0 9275 Unknown I 114 R0427821 0 106 1 0 9275 Unknown I 135 R0427822 - 1 0 9275 Unknown 1 133 R0427823 1 0 9275 UnknOWn I 127 R0427824 I 0 9275 Unknown 50 3 2 5 R0505812 0 126 0 123 0117 I51 0.117 I 0 9275 Unknown SO 18 5 R0505813 I 78 I 0 9275 Unknown SO 4 1 3 RO505814 I91 I 0 9275 Unknown SO 476 R0505815 221 - I 0 9275 I 0 9275 R0505816 I87 186 R0505819 3 01 I 0 9275 R0505820 3 63 I 0 9275 Unknowo RO505821 2 65 I 0 9275 UnknOWIl 74 7 RO505822 3 47 1 0 9275 UnknoUn 76 4 R0505823 3 54 3 26 - I 0 9275 Unknown 250 8 4 0 R0505826 195 I 0 9275 unknown 250 102 R0505827 23 6 I 0 9275 unknown 250 9 9 5 R0505828 231 I 0 9275 Linknow 250 8 4 2 RO505829 195 - 1, 2 5227: ?.I'"III? 22 < I 0 9275 ROSOS833 27 6 I 0 9275 R0505834 33 7 I 0 9275 R0505835 24 3 - 1 0 9275 1 0 9275 1 0 9275 I 0 9275 I 0 9275 I 0 9275 unknoun - I 0 9275 I 0 9275 R0505836 31 9 R0505837 32 0 299 IR0505840 51 5 R0505841 65 4 R0505842 536 RO505843 R0505844 R0505847 I 0 9275 Unknaxn R0505848 1 0 9275 LlnknOwn R0505849 - 1 0 9275 Llnlowwn 500 200 ROSOSSSO I 0 9275 Llnknoun SW ROSOS8Sl I 0 9275 Unknown SW 404 R0427827 I 0 9275 Unknown 1000 294 R0428814 273 I 0 9275 unknown Iwo 293 R0428815 272 I I 0 9275 rnknown 500 R0427830 223 - I 0 9275 Unknown 5W 406 R042783I 188 1 0 9275 Unknown IOW 330 R0428820 306 I 229 I 0 9275 UnlOlDwn 1WO R0428821 270 I 0 9275 llnloloun IWO 417 R0428822 387 I I - 0 9275 0 9275 Unknown L'nknonn 1000 451 R0428823 loo0 435 R0428824 419 403 357 Sample quantitaledoutoflinearrange olcune Date Enleremy Dale VenBedlBy Punty EnteredlVenBed 12/061W KIH lZiM1W HOJ i0413O~Ol LAC 12127100 LAC PFOS 04/27/98 Lot 215 04128198 Lot 215 05105198 Lo1 215 PFOSA R042780263, R043080344 R04278 13-24 R04308 14-25 RO430828-39 RO430842-53 RO42784147.52-53 R04278 150-51 R042785657 RO430856-57 RSD Sld DCV. MYMSD RPD NA NA 3% 96% 53 0 00215 7 10 0 W828 136 0 252 12 7 0415 PFOSA 04127198 Lo1 L-2353 04130198 Lot L-2353 PFOSAA R042780243BSYM R04278 13-24 R05058 12-23 RO505826-37 R0505840-51 RO427827-38 R042785657 RO505852-53 Lot L-2353 PFOSA Purity Camrrlian FsnW UnknOW unknown UnknOWn unknown NA NA NA NA UnknoWl LlnknOwn unknown UnknOUn L'nknOWn unknown unknown UnknOHn unknoxn unknown LhknOwn L'nknOWl UnknoWll unknown unknown 1lnknOwn unknown LlnknOWll UnknOWn Unknown UnknOUn Unknown unknown PFOSAA 04/27/98 Lot 617 05105198 Lot 617 Dilull~ns 111, 11. Ill 111, 111, Ill 1150. 1/10, 1/50 11250, 1/50. I1250 IISW, IilW. IISW 1/1ooo, 115, 115W IISW Ill,111, I l l 111, Ill. I l l PFOSA Dilution Fnomr I 1 I I I I I 1 1 1 1 I I I I I I I IO 10 10 10 10 10 10 10 10 10 50 50 50 UnknOwn 50 UnknOUn 50 UnknOW 50 12 8 UnknOXn 50 3 84 UnknOWl 50 I : 1 1 ImnknOUll 100 UnknOUn IW ClnknOwn unknown Unknown IW cnkn0,m IN unlmown unknown . IW 100 unknown IW unknown 100 UnknOW 5 UnknOUn 5 I unknown 5 18 6 I Unknoun I 5 I 42 5 UnknOWll 5 unknown 5 Llnknoun 500 LlnknOwn Sno 18 3 L1nkn"Wll 5 65 2 UnknOUn 5 PFOS = Pernuo-mesulfo~tonate PFOSA = Pernuo-tanesulio-Ide PFOSAA = P e r n u O m O C t a n e S u l f D n d ~ ~ ~ ~ t a l ~ Corrected PFOS LOQ (0 0249 uglmL) to include rtd comcllon Caclorr newLOQiroO23l ugimL LACO2119~01 Analytical Report: FACT-TOX-001 LRN-U2103 PFOSA COM. n@mL 0 00 0 W 0 W OW 243 228 295 283 0.W 006 000 059 0 31 0 35 0 83 0 64 0 37 021 255 268 297 389 3 39 5 39 656 732 526 657 140 Filmsme R0427802 R0430803 R0427803 R0430804 R0427856 R0427857 R0430856 R0430857 R0427813 Rob27814 R0427815 ROC27816 R0427817 R0427820 R0427821 R0427822 R0427823 R0427824 R0430814 ~043n815 R0430816 R0430817 R0430818 R0430821 R0430822 ROO430823 R0430824 R0430825 R0430828 Conrrnbmtian of PFOSA u@mLor Y. Rce <LoQ (0.00249 u g i d ) <LOQ <LoQ (0 W249 ugimL) <Log 97% 91% 118% 113% <LoQ (0.W249 ugimL) <LoQ(OW249uglmL) <LCQ(OW249ughL) <LoQ(OW249ugimL) cLOQ (OW249ug/mL) <LCQ (0 W249 u # d ) cLOQ (0 W249 ugimL) CLOQ (0 00249 u g i d ) <log(0 W249 ugimL) <LCQ (0 W249 ugimL) 0 0255 n 0268 0 0297 0 0389 0 03390 0 0539 0 0656 0 0732 0 0526 0 0657 0701 0 668 0611 ::4 I 1 I 2sY no430835 344 IC0430836 123 R0430837 270 R0430838 278 R0430839 192 R0430842 126 R0430843 R0430844 R0430845 I 29 I 72 0614 I35 I39 I 92 I26 I29 242 ?I7 R0430846 2 17 234 R0430849 2 34 300 R0430850 3W 266 R0430851 266 223 Rob30852 2 23 218 R0430853 2 18 208 R0427841 I 04 266 R0427842 133 I I 208 R0427843 I 04 267 R0427844 I34 239 R0427845 I20 476 R0427849 2 38 6% R0427835 3 48 126 R0427836 6 29 382 R0427852 I 91 233 R0427853 117 FSD S l d DLV. MYMSD RPD NA NA 6% 4% NA NA NA NA 17 7 0 WS49 14 I 0 00874 I1 7 : "I!$ 0 147 65 6 2W 3M EnEExTcvSe-l8i9-r57Ionmental Laboratory Sen Week 4 Rework TOXM)I -sen-?12- I I AI XIS Page 199 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance# 6329-212 SMy. 104 Week DEW Cmmogemcay study wth N m w Range (98 1%) N-Ethyl P c r f l u o m l m e s u l f o n d ouhanal m &IS Pmduct NumkrCTcst Substance) 1-6316 (MFOSEQH) Rat Sem FACT-M-3 0 & FACT-M-4 0 reworked usmg FTS-8-5 I Chick080697and Madeline 041098 MarrLwZ3& 3 I See listing to the nght See Amchmenls See Attafhmenlr See Amchmenls 04/23198,04128198 RWW 04127198,04/28198,04/30/98,05105/98 KJHMOJ 09101~W.12iOliW, 12'041W. I2iO5/00 M M W H Sample Data WEEK4 RAT SERA I GTOUI) I DOSC REWORK Ssmpbx Lo1617 Correction Factor unknown NA NA Dilution I PFOSAA C0"C. n&Id OW OW OW OW 271 252 143 466 13 7 - Filename - - R0427802 R0427803 Ro4278M) - R0427856 - R0505852 R04278 13 ConrrnlrltlOn afPFOSM u & I der % RIC :LoQ (0 W256 ugimL) :I@ (0 W256 nglmL) :LoQ (0 W256 uglmL) 'LOQ (0 00256 ugimL) 108% 101% 57% 186% 00137 - MI." - P F O S M UYd <toQ 3 - 104% - 122% CO"!ml C90732M 43 2 R0427814 0 0432 0 0 mgkg C90725M 37 7 R0427815 0.0377 C90753M unknown 82 4 R0427816 00824 - 78 7 R0427817 449 <04279820 00787 0 0449 - O O S l l C91143F 85 7 <04279821 00857 C91165F 54 4 COO4279822 0 OS44 Group 2 C91169F C91171F C90789M UnknoW 57.2 COO4279823 59 7 31 1 RO505812 0 0572 0 0597 I55 - 0 0604 C90814M 42 7 RO505813 2 13 3 0 m@g C908IN 32 I R0505814 I60 C90833M 109 RO505815 5 46 45 0 R0505816 2 25 I12 R0505819 5 61 - 2 6 0 C91205F 49 3 R0505820 2 47 C91222F 54 2 R0505821 2 71 C91230F CW846M - 37 9 R0505822 190 53 9 R0505823 9 7r' 56 4 R0505826 - 3 07 45.8 R0505827 I1 4 45 3 R0505828 11 3 - 49 5 R0505829 12 4 57 2 ?.%"%E !f? 73 9 KO505833 I8 5 - !3 2 139 R0505834 34 9 C91285F 54 4 R0505835 13 6 C91287F - 74 3 R0505836 I8 6 121 R0505837 30 2 446 RO505840 22 3 - 33 I 448 R0505841 22 4 39 0 R0505842 195 85 3 R0505843 42 6 - 55 5 RO505844 27 8 47 5 W505847 23 8 - 26 9 C913IR 604 R0505848 30 2 C91333F 640 R0505849 32 0 C91345F - 115 R0505850 57 3 51 5 RO50585I 25 8 I23 R0427827 61 3 - 33 8 Rgh Dare C91W5M 137 R0427828 68 4 3W m g k g I C91024M C91033M 151 R0427829 75 6 126 R0427830 63 I - 75 8 R0427831 37 9 175 R0427834 87 5 - 61 2 C91427F 168 R0427835 83 9 C91430F 1% R0427836 978 C91443F C91448F 271 115 - R0427837 R0427838 136 57 6 - 92 5 * Sample quaniltaredout o f imar range of c u m RSD Sld. Dlv. MYMSD RPD NA NA 7% 106% 56 8 0 0291 25 2 00152 627 I 63 47 3 174 2 3: 38 7 8 95 34 5 9 28 401 13 6 23 2 14 2 30 6 % 1 Date Enternmy Dale VenfieediBy Punty EnterndNenfied 12106!wKJH 121061W HOJiOU30/01 LAC 12127100 LAC Corrected PFOS LCQ (0 0249 ugimL) UJ lncludcstd cometion facton neu'LOQa00231 ugimL LAC02119101 3M EEEnTxScve-lSi9-7r5Ionmental Laboratory Analytical Report: FACT-TOX-001 LRN-U2103 Page 200 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance# 6329-212 Slvdy Pmduct Numbeflest Substance) Mamx MeUlodiRe\alon Analylical Equipment System Numkr Instrument SoRWareNenK," Filename R-Sqquared Value Slope Y-intercept. D a m of E~mctloniAnalpt Dales of Analysis/Analyst Dale of Data Reductiodhlyst Sample Data E E K 4 RAT S E R A R E W O R K MethodBlk Qc-250 ppb Group 1 C0"UOl 0 0 mgng Group 2 Low Dore 30m a g Group 3 Mid Dose 30 0 mgkg Group 4 Mid-High Dose 100 m a g GrrJYP 5 High Dose 300 mgkg Rat Serum Blk-2 MS 4/23/pJ MSD 4/23/98 MS 4/28/98 MSD 4/28/98 C90719M C90732M C90725M C90753M C90775M C91135F C91143F C91165F C91169F C91171F C90789M CW814M C90817M C90833M C90834M C91198F C912OSF C91222F C9123OF C91237F C90846M 1 C90853M C90866M C90890M CrnZ?!?.! C91251F C91272F C91285F C91287F C91291F C90935M C90939M C90942M C90%1M cm%m I C91316F C91317F C91333F C91345F C91350F C90984M C91005M I C91024M C91033M C91037M C91423F C91427F C91430F C91443F C91448F 104 Week D E W Can;mogcrucltyShldy with N m w Range (98 I,% N-Et)hyl P e r f l u o m m e s u l f o mE~thanol m Rats T-6316 (UFOSEOH) Rat S e m FACT'-M-3 0 & FACT-M4 0 reworked using mS-8-5I Chick080697and Madcline 041098 MarrLynr 2 3 & 3 I See attachment^ See Attachments See Attachments See Attachments 04/23/98,04128198 RWW 04127i98,n41~8198,0413019085,105198 KJWHOI n901100, iziniiw, I ~ W W1, 2 m t m MMWH concrntntion Of PFOS uYmL or *h Rcc .. ctoQ (0 0231 u&L) <LoQ 96% 93% 95% 65% 183% * 124% 0 0244 0 0302 0 0244 0 0743 0 0495 0 123 0 117 I51 I78 I91 2 21 I87 3 01 3 63 2 65 3 47 3 54 19 5 1 i!236 23 I I9 5 22 < 27 6 65 4 53 6 72 8 I 58 4 I 58 6 ' 797 I 972189 50 5 188 273 272 I 223 188 306 270 387 419 403 0117 I86 3 26 1 2! 5 299 604 I I 70 7 I 229 357 RSD Sld Div. 00215 n 252 - ! 128 3 84 I4 5 88 CO"CC"trsti0" of PFOSA 97% 91% 118% ~ L C Q(n 00249 ugimL) cLOQ (000249 ugimL) <LCQ(OW249ugimL) < L o p (0 00249 ugimL) <LOQ (0 00249 ugimL) <toQ(0 00249 ugimL) CLOQ (0 00249 UgimL) 0 0255 0 0268 0 0297 0 0389 o 03390 0 0539 00656 0 0732 00526 o 0657 0 668 0611 1 K4 ! 0 680 "132 I I 29 I39 I92 I26 I 2 42 2 17 2 34 MIXll PFOSA UYmL RSD Sld Dtv. MYMSD RPD 'toQ NA <LOQ NA 94% 6% 116% 4% NA <toQ NA NA <LoQ NA 00310 17.7 0 02549 0 0622 14 1 0 00874 15?$ I1 7 "$!$ 31 7 I27 n 404 28 7 I81 0 520 13.8 2 48 0 344 I 04 18 6 I34 I2 3 42 5 I 20 119 0 147 2 38 3 48 6 29 I8 3 I91 65 6 65 2 I17 304 200 FOS = Perflu Conrrntrntion af PFOSAA vYmL or % Rce lIX2(0 00256 ugimL) LOQ (0 00256 ugimL) (0 00256 ugiml.) LOQ (0 00256 ugimL) 108% 101% 57% 186YO 001372 n 0432 0 0377 0 0824 0 0787 0 0449 0 0857 0 0544 0 0572 0 0597 I55 2 13 IM 546 2 25 5 61 2 47 2 71 190 2 70 14 I 11 4 II 3 12 4 :SI I8 5 34 9 13 6 18 6 30 2 22.3 22 4 I9 5 42 6 27 8 23 8 30 2 ' 320 57 3 25 8 61 3 68 4 75 6 63 I 37 9 87 5 83 9 97 8 136 57 6 Analytical Report: FACT-TOX-001 LRN-U2103 - MIX" - PFOSAA WmL <LoQ RSD MYSMldSDDIRVP. D NA CtOQ NA - I 04% - I 22% 7," 106% - O O S l l 56 8 00291 - 0 0604 25 2 0.0152 62 7 - 2 M I63 - 47 3 3 07 I45 - :3 3 - 23 I 38 7 90 34 5 - 26 9 93 - 33 8 401 13 6 23 2 - 61 2 I4 2 - 92 5 30 6 28 3 Dale EnInteredIBy Date VenfieUBy Punt) Entere&Venlied 12'06100 KJH l2ION00 HOJ! 04'30,Ol LAC 12127r00 LAC Comcled PFOS LCQ (0 0249 ugjmL) Io include itd c ~ n e ~ t mfanston n e w ~ ~ e n o :u3g ~i m ~L A C O ~ ~ I ~ O I 3M EnEETxcvSe-l8i9-5r7Ionmental Laboratory Sera Week 4 Reu'oh TOX-001 -rera-2 12-11AI XIS 533 liP2MaI ge 201 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance# 6329-212 Study Roduct Number(Ten Subrtancc) MauiX MethodiRnisioo Analytical Equipment System Number: h m c n t SaflwueNersion Filename R-Squarrd Value: slope. Y-Intercept: Date of ExuactioniAnalyst. Date of AnalyririAnalyrt Dale of Data R e d ~ c t i o n l h d y s t Sample Data WEEK 8 RAT SERA REWORK 104Week Dietary Cueinageniciry Study with ~ a m Rwmgc (98 1%) T-6316 (ElFOSEqH) Rat s m FACT-MJ 0 & FACT-M-4 0 rcworked using ETS-8-5.1 Madeline 041098 M a r a L w 2 3 & 3. I See lining to the "&I See Atllchmmtr See Attachmans See AtWchmenCI 05/11/98,05/14/98 RWWiOK 05/13/98, 05114198 HOJiKJWLAWJJ 04/10/W0.1/03/01 CSWMMH N-Ethyl Pefluomoctmcmlfonamido Ethanol in Rats Lo1215 Filnumes Bllu Grp 1 Grp 5 MS, MSD pms 05114198 LO1215 PFOSA 05114198 Lot L-2353 Analytical Report: FACT-TOX-001 PFOSAA 05/14/98 Lo1617 LRN-U2103 PFOS RO5149802-03 & 8 6 8 7 RO5149812-23 RO5 149826-37 PFOSA R0514980263 & 8687 R05149812-23 R05149854-65 PFOSAA RO5149802-03 & 86-87 R05 149812-23 RO5 149840.5 I Dilotianr 111. 111, Ill lISW0, 1/15. l/SW ROS 149883-84 R05149883-84 RO5 149883-84 111. I l l . 111 Lot L-2353 Conrcntration of PFOSA MIZll PFOSA EL: I I i I 0.0mwkg ~~ C90717M C90718M C90737M C90756M I C90769M I C91126F I C91140F I C91145F I C91162F I C91163F I croap 5 C90971M I _ _ High Dose C90972M I 3Wmgike. C90988M I C90996M I C91017M C91393F I C91394F I I I I C914MF I C91415F I C91438F I Tentative results, CCVs did not meel criteria LAC 01/08/01 Date EnarcdiBy. 05/24/00. 01/08/01 CSWLAC Date Verified/ By 12/28/00 HOJ / 01/08/01 KJH /04/30/01 Purity EnteredNcrified. 12127IW LAC 09275 i;::09275 0,9275 09275 09275 0.9275 09275 09275 09275 09275 09275 09275 09275 09275 09275 0.9275 09275 09275 LAC Corrected PFOS LOQ (0.00976 ugimL) lo include n d correctionfactom new LOQ is 0 00905 ugimL LAC 02/19/01 I I Unlmown Uhwn Unknown Unlmown Unknown Ubwn Unknown Unlmown Unknown Unlmown Unlmown Unlmown Unknown I Unlmown I UnLnown Unlmown Unlmown Unlmown i I :;: I I 369 R05149812 RO5 149813 R05149814 I 871 R05149815 I 495 R05149816 I 1% R05149819 I 183 R05149820 I 176 R05149821 I 225 R05149822 I 194 R05149823 5000 736 R05149826 5000 671 R05149827 5000 736 R05149828 5000 705 R05149829 5000 903 R05149830 5000 I 120 I R05149833 I 5000 I21 R05149834 5000 114 R05149835 5000 130 R05149836 5000 136 R05149837 0 0342 0.M82 OM95 0.0809 0 0459 0.182 0 170 0.163 0 209 0.180 341 311 341 327 419 557 559 530 603 63 I 103X I I I 1 unknown Unknown Unknown I ::E I <LOQ (0.101069%78 UgimL) 110% R05 L498L2 R05149813 <LOQ (0 00978 ugimL) 6.12 R05149814 <LOQ (0.00978 ugimL) 33 5 UnlmOWn I 5.91 RO51498I5 <LOQ (000978 ugimL) 00518 0.0174 unknown I 0.00 R05149816 UnlmOWlI I 6.49 R05149819 <LOQ (0.W78 ugimL) UhOW I 6.47 R05149820 <LOQ (0.00978 UgimL) UnloloWa I 5.92 R05149821 <LOQ (0.00978 ug/mL) 9 60 UhW I 6.W R05149822 cLOQ (0 00978 ugimL) 0 181 0 0174 UDknOWn 1 6.12 R05149823 UhOW 15 76.7 R05149854 U"ln0Wa I5 70.4 R05149855 UnlmOW I5 41 8 R05149856 II 9 UlIkXlOWn 15 88 5 R05149857 348 41 5 LJ"knOw0 I5 88 4 R05149858 I I I unlmown I I5 I 51 5 I R05149861 U"knOW0 I5 113 R05149862 0.773 I 69 UnlmOWn I5 58.8 RO5149863 0.882 6.96 unlmown I5 55 4 R05149864 0 830 576 40 I unknown I5 80.2 R05149865 1.20 PFOS = Peduomactanemlfonatc PFOSA = Perfluorooctancsulfonamide PFOSAA = Perfluomoctanesulfonamidoacctate ETS-8-5 I 3M EEnxcveli9r7 onmental Laboratory S a a Week 8 Rework TOX-01-ren-212-1 IAI XIS Page 202 3M Medical Department Study: T6316.1 RSD Std. De". MYMSD RPD Study: Roduct Numba(Tcst Substance) Matrix McthodiRcvirion Analytical Equipment System Number I m m e n t SoRwveNcrrion Filename. R-Squared Value Slope. Y-lnl~epl: Date of E x u a e l i o d h l y a : Date of Analysir/Andyst. Dale of Data ReducUodAoalyst- Sample Data WEEK 8 RAT SERA REWORK ClOltp Dolt Sample # HZO Blk-1 AMDT# 092597.1 Covance# 6329-212 I04 Week Dietary Carcinogeoiciry SNdy with Narrow Range (98 1%) N-Ethyl PcrfluorwclmesulfonamidoEulvlol in Rats T4316 (EtFOSEaH) Rat Senun FACT-M-3 0 & FACT-M-4.0 reworkedusing ETS-8-5 I Madeline 041098 MassLynx 2 3 B 3. I See listing to thc right See Auaehments See Attachments See Anaehments 05111198.05/1498 RWWIOK 05/13/98, 05114198 HOJIKIWLADIJJ W10100,01103101 CSWMMH Analytical Report: FACT-TOX-001 LRN-U2103 Lot 617 PFOSAA Pu"ty PFOSAA PFOSAA Correction Dilrlio. Corn. FldW Unh0wn Fador n#mL I I I Filensmr Concentration of PFOSAA m@mLor % R s Meall PFOSAA m6/nL RSD Std. DN. MYMSD RPD I NA Rat Sown Blk-2 Unhown I 000 R05149887 <LOQ(OOl00ug/mL) ** <LOQ QC-75 ppb MS 511 1198 NA I 76 9 R05149883 103% ** 12% MSD5111198 NA 1 78 3 R05149884 104'4 ** 103% CrooD 1 CW717M Unlmown I 186 R05149812 < L O O.1,O O I W udm I L) I CanVOl CW718M Ullhwn I 24 3 RO5149813 0.0243 0 0 mg/Lg cw737M Unloloun I 17 9 R05149814 ELCQ (0.0100 u&L) NA C90756M UIlhOWn I 304 R05149815 0.0304 NA C90769M U"lm0W I 24 2 R05149816 E L C Q (0.0100 u&L) 00169 C91126F U"h0Wn I 30 2 R05149819 0.0302 C91140F U"h0W I 55 2 R05149820 0.0552 C91145F U"h0Wn I 38.1 R05149821 0.0381 NA C91162F UllbOwn I 43 4 R05149822 0 0434 NA C91163F U"h0W I 49 5 R05149823 0.0495 0 0433 croup 5 C90971M UnhoWU 500 I51 R05149840 75 3 High Dose CW72M U"hW 500 165 R05149841 82 3 300 mgikg C90988M U"bW 500 103 R05149842 51.6 26.2 C90996M Unlm0Wn 500 126 R05149843 63 2 0 288 C91017M U"bW 500 125 R05149844 62 3 67.0 C91393F UllhOW 500 212 R05149847 106 C91194F UnhlOW 500 130 R05149848 648 C91404F U"h0W 500 157 R05149849 78.7 35 4 C91415F UIIlmoW 500 158 R05149850 79 I 0381 C91438F U"h0W 500 152 R05149851 75.9 80 9 ** Tenlalive results,CCVr did not meet criteria LAC 01/08/01 NA 2% 57 5 0 00974 22.5 0.00973 17.9 12.0 18.8 15.3 ETS-8-5 I 3M EEnxcvel 9i7ronmental Laboratory Sera Week 8 Rework TOX-WJI-sera-212-1IAl xlr Page 5131/2001 203 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance# 6329-212 Study. I 0 4 Week Dietary Clrcinogcnicily Study with Nmow Range (98 1%) N-Elhyl Pefluomlmerulfonunido Ethawl in Rals Product Nwnbm(Test Substance) T-6316 (ElFOSEaH) Matrix. MethdRwision. Rat Serum FACT-M-3 0 & F A C T - M ~0 reworked using ETS-8-5 I Analy6ical Equipment System Number Madeline 041098 l m m e n t SoRwvcNenion MasrLynx 2 3 & 3. I Filename. See Auachmcnls R-Squared Value See Attachmenls slope. See Attachmenls Y-htoccpt See Atfachmenls Date of ExtractiodAnalyrt 05111198.05114198 RWWIOK Date of AnalysiriAnaly~. 05113198, 05114198 HOJIKIWLADIJJ Date of Data ReductioniAnalyst: 04110100. 01103101 CSWMMH Samole Data iEEK 8 RAT SER Gr0.p DO% tEwoRK Sample I Method BIk M a k x Blk H20 BIk-1 HZO BIk-2 Rat S- BIk-1 Rat Senun Blk-2 Concentration of PFOS mdmL or % Rec <LOQ (0 m 5 ugiml) 4LCQ (0 00905 u mi) <LOQ (0 W905 ugiml) <LOQ (0 m 5 u&l) ** cLOQ RSD I Coneratrntion I M a n I Std. De,. 01 PFOSA PFOSA MYMSD RID mI/mL o r % Rw .I/nL <LOQ ( 0 00978 ug m L j UA <LOQ1ocQ978ugmL1 cLOQ cLOQtO 00978 u g m L ) NA <LOQ (0 00978 UvmL) <LOQ RSD Sld D n MYMSD RPD NA NA QC-75 ppb MS 5111198 MSD5111198 104% 125% '8 115% 18% 103% Group 1 C90717M 0 0342 <LOQ (0101069%78 ugimL) lI(p/o 12% COnUOl 0 0 mflK C90718M C90737M C90756M C90769M 0 0809 <LOQ (0 00978 ugimL) <LOQ (0 00978 u d m L ) 33 5 <LOQ (0 00978 u&L) NA 00518 0 0174 <Log (0 00978 UgimL) <LOQ NA 1C91126F C91140F C91145F C91162F C91163F <LCQ (0 00978 u&L) <LOQ (0 00978 ugimL) 9.60 <LOQ (0.00978 ugimL) NA IO181 0 0174 cLOQ (0.00978 ugimL) <LOQ NA C90971M 1.15 C90972M 1.06 C90988M 0.63 11.9 I33 26.2 C91017M 41.5 1.33 110 0 288 C91393F 0 773 C91394F 1.69 C91404F 0.88 C91415F 6.96 0.83 35.4 C91438F 576 40 I 1.20 1 08 0 381 id not mcct criteria LAC 01108101 5124/00,01108101 CSWLAC Date VerifiediBy- 12128100 HOJ I01108101 KJH I 04130101 LAC Purity EntcrdVerified 12127100 LAC I Concentratio. of PFOSAA mdmL or % Ret < L O Q r O 0100 ug m L j ~ L O Q \ O O I W u y m L j* <LOQ,ool00"amLI <LOQ (0 os00 u&L) *' 103% <LOQ (010140%0 ugimL) 0 0243 <LOO IO 0100 W m L ) 0 0304 <LOQ (0 0100 ugimL) 0 0552 0.0381 0.0434 0 0495 75.3 82 3 51.6 63 2 62 3 106 648 78.7 79 1 75 9 Analytical Report: FACT-TOX-001 LRN-U2103 MIS. PFOSAA mI/mL <LOQ <LOQ 103% mn Std. D N . MyMSD RPD NA NA 2% 0.0169 57.53 0 00974 + 00433 22.5 0.00973 18 8 80 9 15 3 Corrected PFOS LOQ (0 W976 ugimL) to include ndcorrection faclors new LOQ is 0 W905 ugimL LAC 02119101 ETS-8-5.1 3M EnExcveli9r7 onmental Laboratory Sera Week 8 Rework TOX-001-rm-ZIZ-l IAl XIS Page 5131/2WI 204 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance# 6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 Study 104 Week Dietary Caninogcnicily Study with Narrow Range (98 1%) N-Ethyl Poduomoclanesulfonamido Ethanol in Rats Roduct Numbm(Ten Subnance) T-6316 (ElFOSE-OH) Matliix Rat Serum MethodiReviSiOll FACT-MJ 0 & FACl-M-4 0 reworked using ETS-8-5.1 Analpical Equipment Synem Number. Madeline 041098 I m m e n t SoftwarcNcnion MasiLynx 2 3. 3 I, 3 2. 3.3 Filename See listing to the right R-Squared Value. See Attachments Slop: See Attachments Y-lntcrcep see Attachments Date of E x ~ a c t i o n i h a l y n . OS11 1/98 RWW/OK Date of AnalysidAndysI 05114198,051i5198.05/17/98.6105198 HOJIKJH Date of Data RcductioniAnalyn. I2/01/W, I2129IW, 01102101 MMWCSH Smmple Data 1EEK 14 R A T S E R A REWORK LolZl5 Grwp Simple A Extndion PFOS Std PFOS Purity PFOS PFOS Filename Concentration MIl" DO% VoI. Rplio Corrcctioa FlClOr Corrraion Dilution Canc. of PFOS mglmL or % RK PFOS o%/mL Mnhod BIk HZO Blk-I H20 Blk-2 I 0.9275 I 0.9275 <LOQ (0.0230 ugimL) <LOQ (0.0230 UgimL) <LOQ Matrix BIk Rat S m Blk-l I Rat Serum Blk-2 I 0.9275 0.9275 <LOQ (0.0230 ugimL) <LOQ (0 0230 ugimL) <LOQ QC-75 PF+ MS 5/11/98 i NA MSD 511 1/98 I NA 104% 125% 115% Group 1 C9073 IM 1 0.9275 0 0842 CiWvOI C90746M I 0 9275 0 0883 0 0 mg/lrg C90748M C90768M I 0.9275 1 0.9275 0 0753 0.0720 C90780M I 0.9275 00779 C91129F I 0.9275 0.148 C91155F I 0.9275 C91160F I 0.9275 60598029 0215 C91174F I 0 9275 UhOW 60598030 0.223 C9llSlF I 0.9275 UhOWn 0 168 Group 2 C90797M I 0.9275 Low Dose C90807M I 0 9275 5 69 3 0 mgikg C90818M I 0.9275 7.14 C90831M I 0 9275 6 53 C90836M I 0.9275 5 85 C91192F I 0 9275 51598054 11.5 C91223F C91233F I 0 9275 1 0.9275 I 51598055 12 0 165 51598056 15.3 C91241F I 0.9275 I1 8 croop 3 C91248F C90843M 1 0 9275 1 0 9275 I 0.9275 1 0 9275 I 0 9275 : I :::;:::: I 10.5 47 0 63.3 47 0 I53 51598033 71.0 I 0.9275 Unhoown 500 145 51598034 67 I 1 0.9275 U h o ~ 500 260 51598038 I20 I 0.9275 Unknoown 500 158 51598039 73 I 1 0.9275 U h o ~ 5W 248 51598040 I I5 I 0.9275 I 0.9275 104 I 0 9275 I 0 9275 I 0 9275 1 0 9275 1 0.9275 192 I 0 9275 I 0 9275 I 0 9275 I I 0 9275 0 9275 Unhoun I I 0 0 I 268 I 51598026 I 248 268 ne For all si ler and nani the initial volume is q u a l to the fmal volumefor an extraction volume ratio of 1. Date EnteredIBy Date Verified, By 6/19/98 LAC, I2106/0C WH. OllO5ll 7/16/98 GML I 12107lW HOI I O M LAC I KJH/O4/3OIOI LAC PFOS = P e m u o r ~ m e ~ i f o n a t e PFOSA = Pduomoetanesulfonamide Purity EnterWerified. IZl27lW LAC PFOSAA = Pduomoctancsulfonamidaacctate RSD Std.Dev. MyMSDRPD NA NA 18% 10 1 OW791 0 0361 192 74 5 25 4 12 8 24 6 12 7 34 0 Filenames Blks Grp 1 G v2 G v3 G v4 MS, MSD PFOS 05/14/98 Lo1215 05/15/98 Lo1215 06/05/98 Lot 215 PFOS 51598004.5. 123.124 60598020-31 51598046-58 51598030.42 51598014-26 RO5149883-84 Lot L-2353 PFOSA Purity Correction Fador UnlmoW U"l0laWn UhOW U"h0WIl NA NA U"lm0Wn UhOW UllhOWn UhOHm UhOW PFOSA DIInUan Fador 1 1 1 1 1 1 1 1 1 1 1 1 I1"h"un UhOWn UhOW U"h0Un UnLnOWn UhOW UhOW UhOW UhOW UhOW UhOW UhOW 5 5 5 . 5 5 5 5 5 5 5 5 PFOSA 05/14/98 Lot L-2353 05117198 Lot L-2353 06/05/98 Lot L-2353 PFOSAA 05/14/98 Lo1617 05/15/98 Lot617 06/05/98 Lo1617 PFOSA 51798004-5.97.98 60598020-31 5 1798016-38 51798043-64 51798069-90 R05149883-84 PFOSAA 51598004-5, 123-124 60598020-31 51598103-116 51598084-97 51598065-78 Dilotiow lil, lil, 1/1 l / l , 1/1, I l l 1/1W.l/2, 1/25 11500, 1/5, l / l W 111o00, 115. li250 RO5149883-84 l / l , 111, 1/1 PFOSA C0"G Filename canerntraian of PFOSA 75 4 84 6 7 65 OW 0.00 744 7.30 OW OW 8.00 OW 7 13 37 0 25.8 29 8 23 4 33 4 46 5 54.0 78 5 53 0 42 6 6.50 124 74.5 128 101 168 117 189 160 142 274 408 397 285 R05149883 103% R05149884 116% 60598020 <LOQ(0.0978 ug/mL) 60598021 cLOQ(0 0978 ugimL) 60598022 <LOQ(O 0978 ug/mL) 60598023 <LOQ(O 0978 ug/mL) 60598024 <LOQ(O 0978 ugimL) 60598027 <LOQ(O 0978 ug/mL) 60598028 <LOQ(O 0978 ugimL) 60598029 <LOQ(0.0978 ugimL) 60598030 <LOQ(0.0978 ug/mL) 60598031 <LOQ(0.0978 ugimL) 51798016 0.0740 51798018 0 0516 51798020 0 0597 51798022 0.0468 51798025 0.0667 51798030 0.0930 51798032 0 108 51798034 0.157 51798036 0.1059 51798038 0.0852 51798043 0 330 51798045 0621 51798047 0 372 51798049 0m1 I I 57198051 51798058 51798060 I 5!798%2 I 51798064 1 51798069 I 5 I79807I 0 505 :;;; 09945 0 799 0 708 51798073 51798075 57198077 0 753 5 I798082 137 51798084 204 51798086 199 51798088 I 42 57198090 2 59 Corrected PFOS LOQ (0.0248 u&L) to include nd Comation factors new LOQ is 0 0230 ug/mL LAC 02119/01 ** Tentative rmlts, CCVr did not m e t criteria LAC 011081Ol Mean PFOSA a@mL <LOQ <LOQ 110% <LOQ <LOQ 0.0598 0. I10 RSD Sld. De". MyMSDRTD NA NA 12% NA NA NA NA 18 4 00110 25.5 0 0280 ETS-8-5 1 3M EEnxcveli9r7 onmental Laboratory Sera Week 14 Rework TOX-WI-sera-212-1 IAI xls Page 513112WI 205 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance# 6329-212 Shldy. RoductNumber(Test Substance) Matrix MeIhcdlRevirion: halyiical Equipment System Number I m m e n t SoflwarclVmion. Filename. R - S q u d Value: slope. Y-laemcpt Date of Extractionihdyrc Date of h a l y s i d h d y n : Date ofDaIa Reductionihalyn. Sample Data I C 4 Week Dietary Carcinogenicity Study with Nmow Range (98 1%) N-Ethyl PerfluomaetanesulfonamidaELhaool in Rats T-6316 (EtFOSEaH) Rat S a m FACT-M-3 0 & FACT-M4 0 reworked using ETS-8-5 I Madelbe MI098 MassLyox 2.3, 3 I, 3.2, 3.3 See lining to the right See Allxchments see Auxhmenfs See AlUehments 05111198 RWWIOK 05114198,05115198, 05/17/98, 6105198 HOJIKJH IZlOllW. 121291W, 01102101 MMWCSH Analytical Report: FACT-TOX-001 LRN-U2103 C91155F U"h0W I 38.0 60598028 0.0380 C9llMlF U"lm0WlI 1 35.9 60598029 0.0359 C91174F UlIlolOwn 1 46 9 60598030 0.0469 C91181F UolmOWn I I8 4 60598031 <LGQ (0.0255 " g i m ~ ) Group 2 C90797M UOlmoWll 25 65 4 51598103 I 63 L O W Dose C90807M UolmOUn 25 28.5 51598104 0.711 3.0 mgkg C90818M UilblOW 25 69 5 51598105 1.74 C90831M UOlmOull 25 55 9 5 I598 106 1.40 C90836M UOlnoUn 25 27.1 51598107 0.68 C91192F UOlmOW 25 56 6 515981 12 I41 C91223F UOlmOWn 25 1 08 5 I598 I I3 2.70 C91233F UolmOUn 25 990 51598114 2.47 C91241F U"lm0W 25 53 3 515981 I5 I33 C91248F UllblOWn 25 57 6 51598116 1.44 Grollp 3 C90843M U"lm0W IW 80 5 SI598084 8 05 Mid Dose C90852M tinknown IW 149 51598085 14.9 300mgikg C90863M U"h0W IW 88 4 51598086 8 84 C90877M Unlm.JWU IW 246 51598087 24.6 C90880M U"h0W IW 126 51598088 12 6 C91281F Unlm0.M IW 106 51598093 106 C91288F UllhOW 100 142 51598094 14 2 C91293F UnlmOWU IW 141 51598095 14.1 C91299F U"h0W 100 309 51598096 30 9 C913MF U"h0W IW I21 51598097 12.1 Groop 4 Mid-High Dose 100 mgks C9090SM C90W7M C90917M C90921M tinknown hh0W U"kn0WU UllkllOW 250 250 . 57 7 99.1 51598065 51598066 14 4 24 8 250 87 7 51598067 21.9 250 97 8 51598068 24 4 C90960M UIlhOW 250 73 5 51598069 18 4 C91329F U"lm0Un 250 102 51598074 25 4 C91337F U"lmO\w 250 153 51598075 38.1 C91355F UDknOW 250 124 51598076 31.0 C9137OF UDknOW 250 158 51598077 39 4 C91379F UnlmOW 250 141 51598078 35.3 Extraction Volume Ratio = lnilial volumdfinal ~ol-e. For d l samples and nandards the initial volume is equal 10 Lhe final volume for an emaction volume ratio of I 0.0363 1.23 187 13 8 I6 4 20 8 33 8 20 9 0.00761 41.1 0 506 35 2 0 659 48.1 6 63 50 4 8.26 21 I 4.38 16 9 5.71 ETS-8-5 I 3M EEnxcvel i97ronmental Laboratory SMa Week 14 Rework TOX-WI-raa-2I2-IIAl XIS Page 513112WI 206 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance# 6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 Study 104Week Dietary Carcinogenicity Study with Narrow Range (98 I%) N-Ethyl Perfluorwclancsulfonaida EUlanol in Rats Product Numbcr(Ter1Substance) T-6316 (EtFOSE-OH) M8ViX Rat Serum McthodiRevirion FACT-M-3.0 & FACT-M-4 0 reworked using ETS-8-5.I Analytical Equipment System Number Madeline 041098 l m m e n t SoftwardVmian. MassLynx 2.3. 3 I, 3 2.3 3 Filcname See Alfachmenls R-Squared Vduc. slow. See Attachments See Anachments Y-intercept. See Attlchmenls Dale of E ~ t r a ~ t i d h d y s t : 05/11/98 RWWiOK Date of AnalyririAnalyn 05/14/98,05/15/98,05/17/98.6/05/98 HOJIKJH Date of Data R e d ~ ~ t i ~ n i A n d y ~ t 12/01/00, 12/29/00, 01/02/01 MMWCSH Sample Data 'EEK 14 RAT SERA REWORK GKUIp I Sample# I Concentration of PFOS mg/mL or % Rec I Meam 1 PFOS RSD 1 Concentration Std. DN. of PFOSA m#mL MYMSDWD u#mL or % Ree I Meam I PFOSA RSD Std. DN. v#mL MYMSD RPD McU~odfllk HZO Elk-I <LOQ (0 0230 UgimL) <LOQ(O 0978 ugimL) H20 Elk-2 cLOQ (0 0230 ug/mL) <LOQ NA <LOQ(O 0978 ugimL) <LOQ NA .. MaUix Elk Rat Serum Blk-1 <LOQ (0 0230 UgimL) <LOQ(O 0978 ugimL) Rat Serum 811;-2 <LOQ (0 0230 ug/mL) <LOQ NA cLOQ(00978 ugimL) <LOQ NA QC-75 ppb MS 511 1/98 104% 103% MSD 5/11/98 125% '* 115% 18% 116% 110% 12% Group 1 C90731M 0 0842 <LOQ(O 0978 UgimL) 0 0 mgng C90746M C90748M 0 0883 0 0753 < W ( O 0978 uglmL) <LOQ(O 0978 ugimL) C90768M C90780M 0 0720 <LOQ(O 0978 ugimL) NA 0 0779 0 00791 <LOQ(O 0978 ugimL) <LOQ NA C91129F <LOQ(O 0978 ugimL) C91155F cLoQ(00978 ugimL) C9116OF cLOQ(0 0978 ugimL) C91174F <LOQ(O 0978 ugimL) NA C91181F 0 168 0 196 0 0361 <LOQ(O0978 ugimL) <LOQ NA Grovp 2 C90797M 5 48 I LOW Dose C90807M 5 69 I C90818M I 7 I4 C90831M 6 53 I I 0 0740 00516 I I 0 0597 II 2 0.0468 I 184 C90836M 5 85 I C~ 9119~2~ F I1 5 C91223F 12 0 C91233F I5 3 C9124 I F II 8 6 14 I I 0 688 I I4 9 0 0667 0 0930 0 108 0 157 0 106 0 0598 00110 25 5 Groq 3 Mid Dose 30 0 m a g C91248F C90843M C90852M C90863M 0 0852 0 330 0621 0 372 0 110 0 0280 I I C90877M 0 MI 28 5 C90880M 0 505 0 494 0 141 C91281F 0 677 C91288F 73.1 C91293F C91299F 130 24 5 C91304F 80.5 1 CW505M 1 220 104 25 4 I I C90907M 0 755 C90917M 0 808 C90921M 0 864 7.78 C90960M 24.6 0 753 0 778 0 0605 C91329F I37 C91337F 204 C91355F 199 I C9137OF C91379F 248 12.7 I42 34 0 2 59 188 :traction Volume Ratio = Initial valumdfinal volume For all samples and s a n d a r k L e initial volume is q u a l to the fmal voIume for an exnaction volume ratio of 1 26 7 0 503 Date Enlercdifly. 6/19/98 LAC, 12/06/00 KJH. Ol/O5lOl LAC PFOS = Pemuo-me~ifonate Date Vcrificdi fly. 7/16/98 GML I 12/07/00 HOJ / 01108/01 KJH / 04130101 LAC PFOSA = Pcrfluomctancsvlfonamide Purity EntcrdNerificd 12127100 LAC PFOSAA = Pcrflua-tancsulfonamidaacetate I Concentration I of PFOSAA w#mL or % Rec <LOQ (0 0255 ugimL) <LOOQQ (0 0255 u&gm/mLL) ) cLOQ (0 0255 udmL) 103% *. 104% ** 0 0539 0 04% <LOQ (0 0255 UgimL) <LOQ (0 0255 ugimL) cLOQ (0 0255 ugimL) 0 0355 0 0380 0 0359 0 0469 <LOQ (0 0255 ugimL) I I I63 0711 1.74 I 1.40 0.68 1.41 2.70 2.41 I33 I :,I 1I 12 6 I 10 6 14 2 14 I 24 8 21 9 24.4 184 I 25 4 38 I 31 0 39 4 35 3 Corrected PFOS LOQ (0 0248 ug/mL) to include std correction facton new LOQ is 0 0230 uglmL LAC 02/19/01 ** Tentative resulls. CCVs did not meet criteria LAC OllO8/Ol Mean PQOSAA m#mL <LoQ <LoQ 103% 0 0360 0 0363 1.23 13 8 16.4 20 8 33.8 I RSD Std. DN. MYMSD RPD NA NA 2% 40 2 0 0145 20 9 0 00761 I I 411 I 0.506 I 352 1I 663 4 38 I 169 5 71 ETS-8-5 I 3M EnExcveli9r7 onmental Laboratory Sera Weck 14 Rework TOX-WI-ren-212-lIM xls Page 5/31/2001 207 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covanceff 6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 Snpk x C91211F C91214F C91221F C9I224F Cll235F C91243F C91244F ".""I'C - PFOS Std 10,171 PFOS Purig - c.rrrrU.n F.dW 0 9275 C.r=t*" F.U., 0 86.0 0 9275 ~ 09275 0 864" am a 09271 ~ 09271 0 8640 (I 8640 09271 ~ HA 0 8640 NA NA NA NA NA NA NA ~ NA NA NA N* NA - NA NA NA NA NA NA NA 0 9275 U 8640 NA (I 9275 U 8640 NA 0 9275 0 864" NA 0 9275 0 86.0 NA 0 9275 0 8640 NA 0 9275 0 86.0 NA 0 9275 0 86.0 0 9275 0 86.0 0 9275 0 86.0 09275 ~ NA 0 9275 0 86.0 0 8640 0 9275 0 86.0 NA 0 9215 0 86.0 NA 0 9215 0 86.0 0 9275 0 86.0 0 9275 0 86.0 0 9275 0 86.0 09275 0 8640 - 09275 0 9275 a 921s 0 9275 0 8610 0 8640 0 86.0 0 86.0 "9215 0 86.0 0 9275 0 86.0 09271 0 8640 0 9271 0 8640 0 9271 0 8640 "927' 0 116". (I 9275 0 8640 09275 ~ U 9275 0 8640 0 8640 0 9275 U 8640 0 927, 0 86.0 09275 0 86.0 09275 0 86.0 0 9171 0 9171 0 9275 0 8640 a 8640 o 8640 ,- 09175 09271 W91. ugh g'mL IAC - 'FOS Sa. - OW ow II I - OW 1.28 - ow 168 NA 216 - NA 166 I54 - I84 NA 37 2 49 2 44 6 40 0 68 5 72 6 55 9 576 50 3 42 2 IIJ llZ 120 997 79 4 98 I 88 I 87 9 - 68 3 92. 274 180 165 5WI 211 408 279 364 343 - 505 412 677 528 430 154 MXI 704 722 603 - 787 ..C.m*",ntb. .~PFOS 4 h L % r(rr LCQ (0 W194 UpimL) LCQ (0 W781 " y d ) UT (0 W394 "&Id) UT (0 W781 " y d ) WQ(OW394udd) LCQ(OW78lUyd) 109% NA 88% NA 67% 62% 2n 71% NA 0 0198 0 0394 0 0357 0 0320 0 05*9 0 0582 OWI8 0 0462 0 0403 0 0338 U 0x88 0114 0- 0 0799 0 0636 0 0786 (I 0706 0 0701 00147 0 074" 2 19 305 I32 401 I69 127 2 2, 2 91 2 75 4 OS 3 54 5 42 4 23 3 44 144 481 5 64 5 7% 4 83 631 FSD Sld E%". MSrmSD RPD NA N.4 NA 21% 10% 22 9 n MI I 21 7 00172 33 0 0 "16 I9 7 0954 UlllnOW" NA NA NA NA NA NA NA NA bobow. uotnorn tinknown Uobown Cnlnovn Uokmorn "akoown liatnorn unlnowo untnowo Unknown Unknown Unkmow ""known U&"Lnorn Unborn Unborn Uoknown Uokmaouo lhknown Unknown Unknown Unkmwn Uuknowll ".k"OWO Uoknown Uoknown L'obom Uoknom Unknown Uoknown U"k"0W" Umkmoxn Unknown lloknown Untnown unt*oom t'"k".."" UObOW llnknom - Sun-.#- - vermd NA HA ~ NA NA ~ NA NA ~ NA NA NA NA NA NA - NA NA NA NA NA NA NA NA NA NA NA NA ~ NA NA NA NA NA NA NA NA - NA NA NA NA NA NA NA NA NA YA NA _N_A _ NA NA NA NA NA NA NA vi( - NA N* - PFOSA C.W. Wid- - OW ow - ow OW - ow ow - 259 NA 242 - NA 181 NA - 202 NA OM om ow OM ow ow ow om (100 - ow (100 (I I R I ow ow ow OW ow ow - ow OW 121 I74 5 53 20 4 126 145 IS 8 176 I, 9 - IS 5 31 0 35 I 37 9 21 4 29 0 29 8 35 8 IS 1 - 43 8 39 6 RSD Sld. Dn. MYMSD RPD NA NA NA 7% 11% NA NA NA NA I00116 3M Environmental ETSI-1 I Excel 97 Laboratory Page 208 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance# 6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 - - - SW"8.k P F O S U PFOSAA VIIW - NA - N A DUuth" F.d.1 - I - I - C.W. ndd ow ~ ow NA NA ~ NA NA ~ NA I I ~ I I ~ I (100 ow ow ~ ow 121 NA NA NA NA NA NA I 1M ROIIIWO21 122% NA NA NA NA NA ~ ~ ~ NA I 2.1 R0317WO22 97% NA NA NA NA NA RTSO2230-MS5.4 - NA NA NA NA - I NA I I - 285 R0327WO23 NA ow nw 115% CW736M NA I 0 5" CW738M CW739M NA I n 28 NA I ow CW717M NA I 2 10 C9075IM CW752M NA I .49 NA I ow CW75JM NA I 304 CW766M ~ NA NA I ~ I 0 I7 ~ ow C91131F NA I 8 07 NA I ow C91137F NA I 0 05 C9111kF UOkmrn NA I ow C9II53F Unborn NA I ow CPllSIF I - - - C9115BF Unkmxi NA UllbOW NA NA NA NA I ow I ow I ow I ow I 170 NA I .5J CWIO2M NA I 217 CW803M NA I 713 CWIOSM NA I 356 OWBMM NA I ,w CW82.M RA I 410 CW821M NA I 6,6 CW83PM NA I 512 N* ~ NA 10 ~ I 180 ~ 578 C91207F Unborn NA I 64, RO32 ?OW76 C91210F Uoborn NA I 663 R0327W77 C91211F Unknown NA I 699 C91214F Unbown NA 10 165 C91221F Unknorn NA I 68. I C91221F C9123JF Unknown NA UllhOUil NA I 685 I 655 UllbDW" - - - " NA I 702 NA I 766 o m r e d PFI P ow74 T NA NA NA q NA NA NA NA NA N* NA I UObOW NA NA NA 'LoQ (0 w977 UgirnL) CLOQ (0 m 7 7 "gld) <LOO 10 w977 "drnL1 RSD Sld. Dn. M W S D RSD NA NA NA 4.5% II% NA NA NA NA 1 3M Environmental Laboratory Page 209 3M Medical Department Study: T6316.1 ,IN81 13047-80 unknow. RTS02210-MSS-3 RTS02230-MS34 RTSQ2230-MS54 CW72IM CW733M W736M C90718M CW7l9M C907.1M C90751M C90112M C9075SM C90766M 01127F C9II28F C91113F C91137F C91144F C9115IF C911SIF C91158F C91161F C91183F CW795M CWIWM CW802M CW803M CW10SM CY"806M C90821M CW825M CW839M CW8.0M C91196F C9l2OiF C9121OF C91211F C91211F CPI22IF C91224F C'II23SF C9124lF C91244F %EF& tintnow - Msl6 - C.W. WhL OM OM ~ ow ~ ow ow _o_w _ 375 272 371 262 323 222 - 350 211 159 OW OW Om OW 7 34 0.330 ow ow OW ~ 0 om OW OM ow ow 2 95 OW OW OW - nw 218 229 202 273 214 211 315 2" 3c6 __3w _ 245 307 323 223 3" 352 2.3 187 - 389 363 & 0 W17. Y AMDT# 092597.1 Covance# 6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 s"rn,.U vIrw NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA N* NA NA NA NA NA NA NA NA N* I* HA WoS miv# F.S I I I I I I I I I I I I I I I I I I I I I I I 1 I I I I I I I I I I I I I I I I . . 'LOQ (0 024%u p i d ) C W (0 0247 u p i d ) CLOQ (0 0248 " g i d ) 67% RO127OW21 107% 2a sox R0328OWI9 83% 2n PI0 R0327OW22 37% R0328OWIB 116 R0327OW23 141 R0328m02I 58% 20 A5.,7%%. 20 OW R0327OW27 'LOQ (0 0247 UymL) OW R0327OW28 'Loo 10 0217 u p i d ) OW R0327m029 OW R03270W30 OW R03270W31 OW R0327W035 OW R0321OW36 OW R0321OW37 NA NA NA RSD Sld. DN. MsiMSD W D NA NA NA 28% 24% NA NA NA NA N* NA NA SA 3M Environmental Laboratory Page 210 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance# 6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 ample Dala 'EEK 27 RAT SEI Gnw %no*# CH1735M C94736M CW738M C90739M C9UIIIM C90751M CH1152M C9VISIM C91117F C91144F C9113JF CPII54F C91158f CW799M C90802M CW801M C908OlM CW8MM CW824M CWPZIM CW839M CW8IOM C91196F C91207F C91210F CII2IIP C91214F C91221F C91224F C9I235P C91243F .* C912UF #".nu& C.rrC","lla" .~PFOS "Z,rnL" x r(rr 'LcQ (0 Do394 UpirnL) . ~ c ~ ( o w 7 8~1 y d ) ' L o g (0 w194 W d ) . W ( O W 7 8 1 ug/mL) :W(oW394Uyd) - ~ o(gow781uwd) 109% NA 88% NA 67% 62% 2n, 7.K NA 0 0298 0 0394 0 0337 0 0320 00549 0 0582 0 0448 0 0461 0 0103 00338 0 En% 0 111 0 097 0 0799 0 06M 00786 0 0706 00703 00547 00740 1 I9 3 05 I32 4 01 I69 3 27 2 23 2 91 2 75 6 05 3 54 5 42 4 23 3 44 4 44 LSl 5 64 518 4 83 RSD Sld. on: MSlMSDRID C.rn"t"t*" of PFOSA ~id.rKRn CLCq (0 w 9 3 u p i d ) 'LCQ(Ow193 UyrnL) 'LCQ ( O W 9 3 WmL) . L C Q ( o w 9 3 rnL) I 104% NA 21.7 0 0172 I~ W ( O w 1 9 3 & L ) C W (0 00493 UymL) 00115 00171 00351 00379 00234 00298 00358 0 0284 RSD sld Do. MyMSD RPD NA NA NA ~ 7% I I% NA NA NA NA 65. OOL16 180 Ow608 yIid.r x Rn .LCQ(00493 "ymL) <LW(00248 uumL) ILcQ(OO493 " Y d ) CLCQ (0 0248 u g l d ) rLoQ (0 0493 u g l d ) 'Lcq (0 0248 uy,"l.) 129% NA 122% NA 97% NA 115% NA CLCQ (0 0248 u y d ) CLcQ(OO2.8 WmL) <LCQ(OO248 yBlmL) CLOQ (0 0248 u p i d ) CLCQ (0 0248 u g l d ) F L C Q (002.8 " g l d ) r L W ( 0 0218 u&L) r W ( Q U 2 4 8UymL) <LCQ(OO248 oglmL) c c q (002.8 "S'mL) j L a g ( 0 02.8 "pi*) .LCQ(oa2.8 U p i d ) 'LCQ(""2.8 "El"&) .LCq("Ol48 "pd) ClCQ(OO248 WmL) CLCQ (00248 U p i d ) cLCQ(OO248 upirnL) rW(00148 upid) ' L c q (0 02.8 " p i d ) CLCQ ( 0 02.8 Y g l m L ) 0 270 0 455 "0 1 3 7 713 0356 0 369 0410 8646 0512 I80 0 578 0641 0 663 0 699 I65 068, 0 685 0 651 0 702 0 766 C."Ce",..l*" SLOQ (0 w977 u p i d ) c w (n w977 ~ y d ) <W(0 00977 u p i d ) CLCQ (0 W977 'd) 50% 31% q - q - 2 2 116% 31% 26% 32% 21 C W (0 w977 UglmL) <Loo10 W977 " d d l I<w(0 w977 u y d ) CLCQ (0 w977 " g i d ) <m10 00977 W d l I E:0214 0299 ~ 0 399 NA NA "0389 161 RSD Sld. E-. MYMSD R I D NA NA N.4 I% 8% NA NA N* NA "23.8 0611 24 6 0 0744 3M Environmental Laboratory Page 211 3M Medical Department Study: T6316.1 AMDT# 092597.1 Cavane& 6329-212 104 Weck Dietar, C-mogenicify T-6316 (EGOSE-OH) Rat sem ETS-84 I & ETS-8-5 I h w y 070799& Ruby 100699 Masislynx 3 3 R 3 4 see Below See Anscbmrots SFF AnaEhmenB See Anarhmenla Study aifh ?+muRange (98 la)S-Elbyl PerflwmoFlancsulfo~doEthanol in Rats Analytical Report: FACT-TOX-001 LRN-U2103 PFOSEA = Perruomactaor Sample # C90847M C908JSM I C90858M 1 C90859M C90864M 1 C90865M C9087iM 1 ~90872~ C90882M cm8m C91252F _I _ I C91254F 1 C91264F I C91267F I C91283F 1 C91286F 1 C91292F 1 C91295F I C91301F C91303F C90916M - I 1 1 C90931M C90937M C90940M C90944M C90945M 1 C90954M 1 c9095m 1 C90957M I C90963M C91312F 1 ~ I C91323F 1 C91328F I C91339F I C91340F 1 C91351F 1 C91354F I C91357F 1 C91369F C91371F - I I imsled PFC lanude M W LCQS are mamidoacetale yl Perfluomoclancsulfonamidoethyl alcohol 2"2) suifonyi ethyl-& S"rr0g.b - vIrinrd NA NA NA NA NA NA NA NA NA _N_A _ NA NA NA NA NA NA NA NA - NA NA NA NA NA NA NA NA NA NA NA _N_A _ NA NA NA NA NA SA NA NA NA NA p 139410W - PFOS Sfi - C o r r a t i o Factor 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 ~ 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 - 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 __ 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 - 0 9275 0 9275 0 W974 u 'gld u lot 171 pmskity comtlon Factor 0 8640 o 8640 0 8640 0 8640 0 8640 0 8640 0 8640 o 8640 0 8640 0 8640 0 8640 0 8640 0 8640 0 8640 o 8640 0 8640 0 8640 o 8640 0 8640 0 8640 0 8640 0 8640 0 8640 0 8640 0 8640 0 8640 0 8640 o 8640 0 8640 0 8640 o 8640 0 8640 0 8640 0 8640 0 86411 o 8640 0 8640 0 8640 o 8640 o 8640 lo i m i d e std E 19/01 Date EntcrdRy Dale Yenfiedl By Punlv EnledNsrificd 0411IIW, 04/Iz/W, 04/171W,06iO3C?, 06/07/W, 0711IIW, 08ilSiW.09/OL/W CSHLAC OY22101 KIH/O4130101 LAC OY19101 LAC - nos - Dilutioa LUW IW IW IW IW IW IW IW IW IW _IW_ IW IW IW IW IW IW IW IW IW - IW m 2m m IW m m 103 IW ?ow IW ~ 1wo - m m m m m m m m 1wo E mrGt PFOS CON. mdmL 183 269 268 305 268 271 281 235 243 213 629 536 391 506 419 587 481 552 520 329 637 429 509 836 523 509 874 624 633 875 122 145 848 88s 736 Fikosme ormos VymL or Y. Rm ~os19wo19 R0519wo20 21.6 R051900021 21 4 ROSi9wo22 R0519wo23 RO51900028 ~os19woz9 R051900030 RO519wo31 R051900032 RO519wo37 50 4 ROSl900038 42 9 R0519wo39 31 4 RO51900040 40 5 RO519wo4l 33 5 R051900046 47 0 R0519wo47 38 s R051900048 44 2 R05190049 41 7 RO5I'X)IMJO 26 3 RO519wo55 973 R051HX)1156 68 7 R051900057 81 6 R0328MM27 67 0 R051900058 83 9 ROSI'XKXI59 81 5 ~0328~032 70 0 ~03.?8~~33 SO 0 A0817MM23 101 R032800035 70 I Do403MM20 I96 W40300021 232 EO40300022 136 LN~O~WZ~ 142 w40300024 0040300029 146 RSD Std. Ikv. MSMSO RPD lot L-I5709 FTOSA hurily C0,reCliOO Surrog.~ vcriri - PFOSA - Dilution F.Cto7 I I - PFOSA - COOr. WmL 215 302 I4 0 20 3 2 85 I 1 I I I 1 I 1 ~ 1 I I 193 239 236 228 204 247 233 - 163 441 446 433 A062600019 A062600020 A062600024 A062600025 A062600026 A062600027 A062600028 A062600032 A062600033 A062600034 18 6 39 7 7 39 197 77 2 IS 2 I 1 I I I - 1 1 1 I I I I I I I I I ~ I I I I t 516 631 490 557 706 - 491 365 424 524 382 476 410 269 442 292 554 - 402 779 863 606 954 % , A062600035 A062600036 A062600040 A062600041 A062600042 A062600043 A062600044 A062600048 A062600049 A062600050 A062600051 A062600052 A062600056 A062Mw57 A062600058 A062Mw59 A062600060 A062600064 A062600065 A062600066 A062Mw67 4"6,'rn6.P i 963 I 896 22 6 170 38 3 I - I IO 558 - 627 174 A062600075 W719wo17 0 239 0 236 o 228 0 204 0 247 0 233 0 163 0 441 0 446 0 433 0516 0631 0 490 0 557 0.706 0491 0 365 0 424 0 524 0 382 0 476 0410 0 269 0 442 0 292 0 554 0 402 0 779 0 863 0 606 0 954 MI.. FTOSA "diUL RSD Std. Ikv. MWMSD RPD 0 226 163 0 0368 o son 199 0 IO1 0418 21 6 0 0903 o 875 38 6 0 338 ETS-8-5 I 3M Eonlccvl9i7ronmental Laboratory sera wee* 27 (2) TOXaOl-~ra-2Il-IIXAII. Page 212 3M Medical Department Study: T6316.1 AMDT# 092597.1 Cnvanee# 6329-212 104 Week Dietary CamlnogeolcilyStvdy M b N m w Range (98 1%) N-Ethyl PerIIuomactanesulfo-amido Ethanol in Rats T4316 (EtFOSE-OH) Rat serum ETS-8-4 I & ETS-8-5 I Davey070799 E Ruby 100699 Marslynx 3 3 E 3 4 see Below Sse AttachmPnIs SFCAffaehmcnts See A l t a c h n l s 02/23/W, 03/28mO SAI 03/07/00,03/27iW, 03/28/W, 04103/W, 04i05lW. 04/06/w,05II9IW. 06'261W. 07/19iW, 08/17/00 IAS!HOI!UMWCSH 03/09'W, 04/03/W,04104/W,04iVSiW. 01/07/00,04i10i00,05123iW. 06/27w), 07/25/W, 08/18/00 IASMOIMMH WEEK 27 RAT SEF *UP Dole omup 3 Mid Dose 30 0 mgkg omup 4 MDd-Htgh Dosc 100 mgkg s.mpr x C90847M C90855M C90858M C90859M C90864M C90865M C90871M C90872M C90882M C90886M C91252F C91254F C91264F C91267F C91283F C91286F C91292F C91295F C91301F C91303F C90916M C9093lM C90937M C90940M C90944M C9094JM CW954M C90956M C90957M C90963M C91312F C91323F C91328F CY1339F C91340F C91351F C91354F C91357F C91369F C91371F 81s - surrq.1. - Filrmme veriw - NA NA 444 ;: I NA 204 NA - R0328K080 R0328W081 R0328wO82 R0328W83 NA R0328wO84 NA R032800087 NA R0328wO88 NA R0328K089 NA 188 R0328wOH) ~ NA NA 139 R032800391 1 39 ~ 378 R032Bwo94 3 78 NA R0328wO95 3 05 NA R0328WO% 2 84 NA R0328wO97 3 26 NA R0328wO98 2 78 NA M6 R0328WIVI 6 06 NA 293 R0328WIO2 2 93 NA 424 R0328WIO3 4 24 - NA 280 R0328W104 2 80 NA 286 2 86 NA IW 991 9 91 NA IW 705 7 OS NA IW 750 7 so NA IW 856 8 56 NA IW 671 6 71 NA 460 4M NA 10 503 5 03 NA 10 342 3 42 NA 10 480 4 80 NA 1w 819 R0328wO35 8 19 ~ ~ NA 100 964 w403wO34 964 NA IW 977 w403wo35 9 77 NA 100 630 Do403wO36 6 30 NA IW 865 W403wO37 8 65 NA 100 535 Do403wO38 5 35 SA Do40lwo41 8 26 NA W401wo42 113 NA W40W3 NA W40lwo44 IW 980 ~40lwo45 -0Qd(0w492 & 0 w974 /mL)laloclu m394 E 0 w781 u g i ~ c 0Yl9'0l la1 uolnown/swI3 M S M S D RPD Faclor NA NA NA 2 27 NA NA NA 3 46 NA NA NA 6 58 NA NA NA NA NA I NA NA NA Fmrlor Analytical Report: FACT-TOX-001 LRN-U2103 COOC. "ghL 0 490 1.00 OW 000 000 0 490 04W I 46 2 41 0 560 000 000 ow 000 4 10 000 5 02 0 930 0 330 000 6 69 6 19 590 6 38 9 47 4 20 4 52 6 76 I7 4 8 79 8 05 14 7 599 134 IL 119 25 6 11 6 3 93 9 06 18 2 Fiknmr Coorcohalloo orEIFOSE-OH A062600020 UCQ(0 0248 u p i d ) A062600024 <CQ (0 0248 u p i d ) A0626W025 UCQ (0 0248 u p i d ) A062600026 aCQ (0 0248 u p i d ) A062600027 cLCQ(0 0248 u p i d ) A062600028 <LCQ (0 0248 u p i d ) A062600032 UoQ(00248 ug/mL) A062600033 <LCQ(VV248U d d ) A062600034 <LCQ(00248 ug/mL) A062600035 <LOp(V 9248 Y d d ) A062600036 <LCQ(OV248u p i d ) A062600040 <LO? (0 0248 u g / d ) A062600041 <LCQ (0 0248 u p i d ) A062600042 <LCQ (0 0248 u g i d ) A062600043 A062600044 CCQ(0.0248 u p i d ) <LO? (00248 Y g l d ) A062600048 <LCQ (0 0248 u g / d ) A062600049 <LoQ (0 0248 U d d ) A062600050 <LCQ (0 0248 u@nL) A0626WoSI UW (0 0248 u p i d ) A0626000SZ CoQ(0 0248 ug/mL) A062600056 UCQ (00248 u u d ) A062600057 <CQ (0 0248 u d d ) aw A062600058 <LoQ (0 0248 u p i d ) ~0626~059 (o 0248 ~ g / d ) AV62MWM Uw) (0 0248 u d d ) A062600064 UCQ(0 0248 u d d ) A0626W065 <CQ (0 0248 U U m L ) A062600066 <LOP (0 0248 u p i d ) A062600067 A"67-m <" UCQ (00248 tl M "?** ug/mL) "&.M) A062600072 0 0156 RSD Std. Dcv. MSIMSDRTO NA NA NA NA NA NA ETS-8-5 1 3M EnEav4i9r7 onmental Laboratory sera week 27 ( 2 ) TOXM)I-ncm-2l2-l IAI XIS Page 213 3M Medical Department Study: T6316.1 AMDT# 09259'7.1 Covancdl6329-212 imple Data 'EEK 27 RAT SEI hUP Dose S.mplcY Chup 3 Mid Dose 30 0 rn&E C90847M C90855M C90858M C90859M C90864M C9086JM C90871M C9087zM C90882M C90886M C91252F C91254F C91264F C91267F C91183F C91286F C91292F C91295F C91301F C91303F CW16u CW93IM C90937M CW940M C90944M C90945M C90954M C90956M C93957M C90%3M C91312F C91323F C91328F C91339F CY134OF C91351F C91354F c91357F C91369F C91371F ,ate ,"amide 104 Week Vmtary Csrcmo~en~cislytudy w t h Narrow Range (98 1%) S-Ethyl Perfluomoclaocnulfo-& Elbanolin Rats T-6316 (EIFOSE-OH) Rat Spm ETS-84 1 81 ETS-8-J I Davey 07079981 Ruby 100699 Maslynx 3 381 3 4 see Below See Aftachmols %e Allschnxnts See Affachnxota UU231W.03128/W SAL 03/07/W. 03/27/W, 03128/W,04/03/W, 04/05iW, 04!06/00, 05,19/W. 06/Z&W.071191W,08/17/W IASHOJIMMWCSH 03/09/W, 04/03/W, 04I04'W. 04/0J/W, 04107lW. 04/101W,01/2?/W.06/27'W. 07125/W,08I18lW IASHOIMMH new L a M M556 Dilutiion Fmrlor 10 10 10 10 10 10 10 I0 10 10 10 IO 10 10 10 10 I0 10 10 10 1W 100 1W IW 100 1W IW 1W IW 1W 100 1W IW IW IW IW IW IW IW IW CQS (0 w492 & I 0194 & Ow781 u M556 C0"C. Oe/mL 207 20s 21 I 186 182 251 167 301 I49 168 385 340 331 480 398 298 550 432 428 314 71 8 84 7 81 7 61 8 69 8 566 73 8 58 4 61 4 40 9 41 3 640 44 I 88 4 51 4 74 0 77 2 43 6 49 6 RUJ1900083 F.051900084 RUJ1900085 RU5 I 900086 RU51900091 A0817ooO24 RU51900093 A0817wO25 RUS190009J RUJI'XXIIW A0817wO26 R0519W102 R051900103 ROS I SWIM R0519WIO5 R05IHx)I 10 R0519WIII R0519W112 R0519W113 R051SW118 RU51900065 R0519wo66 RU51900061 R05Ipwo68 R05 I900073 RU5 I900074 RUJI W 7 5 RU51900076 R05 I900077 RUJ19wo82 w403wo34 00403wo35 W403KM36 W4U3wO37 0040300038 WI030W41 w403wo42 0040300043 Coocenll.fio. of M5M U g h L or x nee 2 07 2 05 2 I1 186 182 2 51 1 67 3 01 I49 I 68 3 85 340 3 31 4 80 3 98 2 98 5 50 4 32 4 28 3 I4 7 18 8 47 8 17 6 I8 6 98 566 7 38 5 84 6 I4 4 09 4 13 640 4 41 8 n4 5 I4 7 40 7 72 4 36 4% 10 2 blOIS RSD Std. Der. MSrmSD RPD - 2 03 22 2 0 449 - 20 1 3% 0 7% 197 6 61 I30 ~ - 33 2 6 36 2 I1 Analytical Report: FACT-TOX-001 LRN-U2103 - SW0g.f. - V l l i f i d NA NA NA NA NA NA NA NA NA - NA NA NA NA NA NA NA NA NA - NA NA NA NA NA NA NA NA NA NA NA NA ~ NA NA NA NA NA SA NA NA - NA NA PFOSEA miution F.CfW 1 I 1 I I I I I I I I I I PPOSEA C0"C. "ghL OW 0 IS0 OW 0 140 OW OW OW OW 000 OW OW OW OW OW 0 620 OW OW OW OW OW 0 510 OW (100 OW 0 170 OW OW UW OW OW OW 0 350 OW 0 490 "M Ow) I 65 0 380 00lW OW FUro.mr A062600016 A062600017 A062600018 A062600019 A061600020 A062600024 AU6ZMW25 A062600026 AD62600027 A062600028 A062600032 A062MW33 A062600034 A062MW35 A062600036 A062MW40 A062600041 A062600042 A062MW43 AU62MW44 A062600048 A062600049 A061600050 A062600051 A062600052 A062600056 A062600057 A062600058 A062600059 A0626000M A062MWM A062600065 A062600066 A062600067 A"h'rn6,ll A062MM072 A062600073 A062MW74 A062600075 A062600076 - M"" - PFOSEA mdmL RSD Sld. De-. MSIMSD RPD NA 5ZL NA NA Nn NA NA NA ETS-8-5 I 3M EnExvceil 9r7 onmental Laboratory sera week 27 ( 2 ) TOXM)I-sera-211-l IAI XIS Page 214 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance# 6329-212 104 Week D e w Carcinogenicity Sludy with N-u Range (98 1%) N-Elhyl Perfluomoftrncsulfom~amidoPhanol in Rats T-6316 (ElFOSEQH) Rat Serum ETS-84 I &ETS-BJ 1 Darey070799bRuby 100699 Masrlynx33B3 4 Ses AffachmroS Sce A l t a c h l s Sa Altacbnls See Allrchmrnts 02/?3/00,03/28/00 S A L 03/07'00,03/?7/W, 03128/00,04/03/00, 04/01/00, 04/06/W, 05i19IW. 06/26/00, 07/19/00.08~17/W IASMOJMMWCSH 03/09/W, 04/03/00, 04104100,04/0S/00,04/07/00. 04110/00.05/23/00, 06/21/00, 07/25/00, 08/18/00 IASMOJMMH Analytical Report: FACT-TOX-001 LRN-U2103 cmvp Done croup 3 Mid D o s 30 0 mekg Smmpk X C90847M C908SSM C908S8M C90859M C908MM C9086SM C90871M C9087zM C90882M C90886M C91252F C912S4F C912MF C91267F C91283F C91286F C91292F C9129SF C91301F C91303F c90916M C90931M C90937M C90940M C90944M C9094SM C90954M C90956M C90957M C90963M C913 I ZF C91323F C91328F CY1339F C91340F C91351F C91354F C91357F C91369F C91371F tc I :::24 4 42 9 31 4 405 33 5 47 0 38 5 442 41 7 RSD Std. I k v . MSlMSD RPD 14 0 2 8s I8 6 7 39 197 IS 2 169 22 6 146 I70 38 3 'mL) to include I c 02/19/01 /" . . ~ ,~.",". .II. .'<. ~* >?.., ~ ~3h#,,C,"., 111"" L "I ,I"_. PFOSEh = Perfuomoclancsulfonyl e t h y l m d s Date Entersmy Date Verified! By Purify EntcdNenIied 04/11/00, 04/12/00. 0 4 / 1 7 / ~0, 6/02/00,06107/00. 07/11/00, 08/15:00. 09/01/00 C S m C 02/22/01 KJH i 04130/01 LAC 02/19/01 LAC coorontr.tioo 01PFOSA eymL or K It0 21s 0 302 0 193 0 239 0 236 0 228 0 204 0 247 0 233 0 163 0 441 0 446 0 433 0516 0631 0 490 0 557 0 706 0 491 0 365 0 424 0 524 0 382 0 476 0 410 0 269 0 442 0 292 0 554 0 402 0 779 0 863 0 M6 0 954 0 7S8 0 963 0 8% 0 5S8 0 627 I74 lXCLlO" faefom FSD Std. De". MSMSD RPD I6 3 0.0368 19 9 0 101 21 6 fl 0903 "38 6 111 C0llCllIf"'b~ of PFOSAA UymL or K llec 2 30 4 44 2 04 2 S6 2 47 2.12 I 94 1 s3 I88 139 3 78 3 os 2 84 3 26 2 78 6 06 2 93 4 24 2 80 2 86 9 91 7 05 7 so 8 56 671 460 S 03 3 42 4 80 8 19 964 9 77 6 30 8 65 5 15 8 26 I1 3 5 SS 5 80 9 80 u#mL MSlMSDRPD 31.3 6 58 2 06 I 8041 51': ETS-8-5 1 3M EEnxcevl 9i7ronmental Laboratory sna WCCk 27 ( 2 ) TOXM)I-wra-2l2-llAXIIS Page 215 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Gm"p Dow Smmpk X Omup 3 CW847M Mid Dose C908SSM 30 0 mgkg CW858M C90859M CW8MM CW865M CW871M CW871M CW882M C90886M C91251F C91254F C912MF C91267F C91283F C91286F C9129S C9129JF C91301F C91303F C90916M CW931M C90937M CW940M CW944M CW945M C90954M CW956M CW957M C90963M C91312F C91323F C91328F C91339F C91340F C91351F C91354F C91357F C91369F C91371F ,mate fonamide 'OSAA = Psrn"omaetanFsvlfonarmdo.Ectab ... .,.", il"l.l(.l ,L.._cyy, EIFOSE = N m r Range N-Elhyl Perflvomoclanesulfonamideothyl ilcahol l.~.,l..l,,ll.,.ll~,.,~. i ~C " Y . . PFOSEA = Perfuomocfancsulfonyl ethylarm& M"0 EtFOSWH udmL <LCQ UCQ 'Lop UCQ rected PFOS new LOQS a n 0.1 RSD Std. Der. MSMSD RPD NA NA NA NA NA NA ;; 1 Eonerolr.tioo dM5% ugimL or K llre 2 47 2.05 2 11 I86 I82 251 I 67 3 01 I49 I 68 3 85 340 3 31 4 80 3 98 2 98 5 50 4.32 4 28 3 I4 7 18 8.47 8 17 6 I8 6 98 566 7 38 5 84 6 I4 4 09 4 13 6 40 4 41 8 84 5 14 !% Mcm M556 u#mL 2 03 3% 6 61 1 636 i (0W492 E 0 W974 u p i d ) to mcludc std wmclim fact Date Enleredmy Date VcnfieU By PunlyEntere&Ve~&d 0411IIW, 04/12/W.04117 Do. OhiOXw, 06107lW.0711 IIW, 081151W,09101lW CSHAAC OMY01 KJtI104130/01 LAC 02/19/01 LAC RSD Std. De". CoOe..t,.tio. olPMSEA UCQ( 000975 ugiml) noq( 0 00975 u p i d ) RSD Sld. Dev. MSlMSD RPD NA NA NA N* NA NA NA NA ETS-8-5 I 3M EEnxcevl9i7ronmental Laboratory sera weer 17 (I) TOXaOl-rrs-ll2-IIAI XIS Page 216 3M Medical Department Study: T6316.1 AMDTX 092597.1 CovrneeX 6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 'OS = RrnuomocUocl - PFOS S t l - 101171 PFOS ?wl - PFOS - 'arrrrtho F.UW 0 9115 C..nrt*, - me1.r 0 8640 - muclan F.UW I 0 9275 0 86.0 I 09275 _ _ 0 9175 0 9275 0 8640 __ 0 %MO 0 8640 I - I I 0 9175 0 8MO I 0 9275 _ _ 0 9275 0 9275 0 8640 _ _ 0 8640 0 8MO I _I _ I 0 9275 0 8640 I 0 9275 _ _ 0 9271 NA 0 8640 _ _ 0 8640 NA I - I I NA NA ~ NA NA NA ~ NA - I I NA NA NA NA - N A NA 1 ~ ~ NA NA NA I NA NA NA I N A ~ N.4 NA NA NA - I HA NA - " - - NA NA NA NA NA NA NA NA NA 9275 0 8640 I NA 0 9175 0 8MO I 0 9275 0 8640 I NA 0 9275 0 86.0 I NA 0 9275 0 8MO I 09275 0 8640 I 0 9175 0 1164 I 0 9275 0 8640 I 0 9275 0 86.0 I NA 0 9275 0 8640 I 09275 0 8640 I 0 9275 0 86.0 I 0 927s 0 8MO I 0 9215 0 8640 L 0 9275 ~ 0 9215 0 8640 ~ 0 8640 - I I I Confimrd High "9275 0 8640 I I C o a f i m d High 0 9275 . I ...~. ".., ".,.^ , "_".......1_.I1~. n 0864 I " "-" " " I CmfimdHigh 9275 8M" I I I NA NA "0 9215 9275 0 8640 0 86.0 I 1 I ConfimdHigh 0 9275 0 8MO I I ConfimrdHigh 0.9215 0 86.0 I 0 11. I ConfimdHtgh 0 9275 01MO I 00301 I ConfimrdHirh 0 9275 0 86&0 I I ConfimdHigh 'I9275 0 8MO I i - - - I NA 0 9275 I ConfimrdHigh 0 9275 I P."..".c..-.1....." ....."I "?,1 I NA 'I9275 0 86&0 I 0 8640 I "649 0 8640 IO I , Y.4 c 92'5 e 8640 IO 0 9175 0 8640 I 0 9275 0 8640 10 I23 "0 9275 9275 0 " 8640 8640 10 10 223 291 __ + 0 '1275 0 8640 I 0 9275 0 1640 I 0 9275 0 8MO 10 0 9275 ~ 0 8640 - 10 153 09275 0 8640 IO 18, j I ;;NA I I NA 0 9275 09275 "9275 0 9275 0 9275 0 8MLl IO 0 8640 IO 0 8640 10 0 8640 IO 0 86.0 in 09275 0 8640 IO 197 R05020W35 4 93 3 68 2 1. 5 84 6% - - - 09271 0 11640 IO 2 91 0 9275 0 8640 IO 6 so 0 9275 0 8640 IO 5 72 3M Environmental ETS8.J I Exai 97 Laboratory - - PFOSA PFOSA - OU",*O F.tC. 1 I - C.n. WmL ow om I - I I I - I I I - 0.00 OM ow 0.w ow ow ow I OW Unknown - I 0.w I - 0.w I 206 - I 216 NA - I I , , NA NA NA NA NA NA NA NA NA NA NA NA NA NA I 132 I - I NA 253 - 275 NA - - NA NA NA NA I I49 I I79 I I45 Unborn I I19 untaorn I I31 Unknorn I I49 I 153 I I36 I ow I 149 I OW I ow0 Unknown I 0 870 - I ow - I I (I. NA I 0 880 I I 0. I Uakmrn Cmfimd L%h I ,: 0 830 1 I010 Unborn NA I 0 950 Unborn NA I 0 970 Unknorn Confimd High I I IO UaXoorn Confi-dLgb I 118 Unhown C o n f i m d Higb I Ow0 Uokmrn ConfimdHigh 1 I18 Umhorn U"h0W U,~n.k,.m...r n Coofimdlgb NA .ll.. Coofimd High -.e. I I - I I OW ow - I 0, 1 io 12 8 1 16 I I I2 3 I 12 8 I I3 3 I I5 2 I I3 3 I 118 I - I I 13 8 - 23 6 28 6 I 34 5 I 32 1 I I92 I 159 I 27 4 I 26 I I 27 4 Lhknarn - I I - 34 5 30 8 Rso Sld. k. .WSDrn NA NA NA 3% 1% NA NA 8% 9% NA NA NA NA NA h* NA NA NA NA Page 217 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covmce# 6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 Dow Mdhd BIk hhmr Blt hhmx Rlk C90727M C90730M C90751M C90752M C90754M C90755M C90758M C90759M CM761M C90766M C90767M C90771M C91121F C91122F C9IIIIF C?!!I!P C'IIIISF CPIIJIF C91112F Confimrd High Conhmrd H i d NA NA t183% 5% C91227F C9121PF CPI23IF C91244F C9125OF Unborn Conhmrd High I Unbow 2ndAndysisOK I Unknown VA I 1 "k".,rn VA I Laknowo VA I 177 I R0524WO24 809 Do421m18 870 680 I 0012loM59 M.1700084 I I77 0809 0870 0680 - 0 597 83 2 0 697 ~ - 0 5 5 6 - 35 7 0 199 NA NA NA HA NA NA NA NA NA NA NA I NA I NA I NA I NA I N* I NA I C o n f i d High I NA I ConhmrdHigh I Confimrd Htgh I C o n f i d High I NA I 2nd AnslyssOK 2ndAnul.lymrOK I NA I NA I C o n h m d High I 2nd Amlyns OK I Conhmrd High I 2ndAndynsOK I Conhmrd High I q ... ....N A I 2nd :.. AnalvnaOK ^" I I NA Comfimrd Htgh C o n f i d High Confimrd Htgb NA NA NA I NA I NA I I etFOSEOH Corn. 2@!L 2 89 0 8M - "oww 2 57 ow _ooww_ ow ow ow ~ ow 78 2 98 3 ~ 95 1 69.5 78 1 ~ 64.9 91 6 115 ~ 114 87 2 - 70 6 91 2 ow OW ow ow OW ow ow ow OW ow ow ow ow ow ~ ow ow ow ow "I", ow ow ow ow ow ow "00 ow ow - ow"" n Y YY ow GW OW ow ow 0 880 ow ow OW ~ ow 4 27 4 49 4 35 2 17 3 29 111 4 78 3 64 - 399 5 *'I c.lu.ntntlon of EIFOSEOH "gid.,Jbll.. 'LDQ (0w977u y d ) CLOQ (0 00977 u y d ) <LCQ (0 w977 u y d ) 'LDQ (0 w 9 7 1 u y d ) t L W (0 02977 u y d ) .Loll (0 w977 u y d ) <Looio w977 U J d I ' L G (0 w971&"Lj rLoQ(Ow977uymL) CLCQ I :E CLDQ I 3M Environmental Laboratory Page 218 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 5mpk# CW761M C9077.M C91121F C91111F C911J2F C9llSlF C91156F C9llJlF C91178F CIlllOF r?!!$?F C90781M CW791M C90793M C'X1798M C90799M C908WM C90812M C90822M C'X1829M CW832M C9119.F C91197F C91159F CI12WF C91206F C91227F C91229F CPI23IF C912b.F C91250F - .8 Sur",.* verlfkd NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA N.4 NA NA Conhmrd High NA CoohmrdHigb C o n f i m d Hi%b Conhmrd High NA NA NA _..... r o n s m dWgh ....... Conhmrd High "_.-"=-~, " ~1 L" Conhmrd High NA NA CoohmrdHigb CoohmdHigb Confimrd High Conhmrd High Conhmrd High NA C o n h m d Hi%h C-35-d H+ Canhmrd High Y* NA NA Conhmrd High 2nd *".ly.l.OY Cmh-d High Canhmrd High Confimd High Conhmrd High Confimrd High C o n f i m d High Comfimrd Htgb C o n h m d Hlgb Coofimrd High Cmofimrd High Conhmrd High NA NA NA - M Y come. om 0 01 OW _ow_ OW ow ow ow ~ ow ow MSUIw02I OW __ O M 116 282 336 NA NA 305 ~ 107 308 329 N* N* - 125 I36 Oil0 OM om OM 0 340 0 450 0 380 ow ow 0 33 ow0 ow ow ow ~ I78 " 82" 0 320 . .".r 0 00 I01 000 0 260 OW 000 I55 0220 ow - 0 J10 !364 344 3'6 189 294 424 429 141 286 416 ~ re, UI 691 521 403 167 418 408 - 476 501' 356 0.03 0 167 0.18 0 408 n 476 0 504 0 156 NA NA NA 9% 11% NA NA 1% 55 NA NA NA NA NA I.'. 29 5 Oil0 30 1 0 134 Sump* \'"nkd NA NA i F.U., NA NA NA NA NA NA NA <LOQ(0 w192 whL) NA < W ( O W492 ug/mL) NA <LOQ(Ow492upid) NA < L o p (0 W492 u p i d ) NA 70% NA 64% NA .6% NA NA NA NA 51% 2nd NA 31% 2nd NA 63% NA 76% NA 78% NA 63% 2nd NA NA NA NA NA NA NA NA NA 1 OM NA 1 ow NA I ow <LOQ(0 w492 U y d ) NA NA NA NA NA C o n h m d Hish NA confimrd High CmhmdHigh Confimrd High NA I OW I ow I ow I ow I I ow ow ow ow ow ::: NA NA 2nd AndgnsOK 2ndAndyniOX ;,,;A,,.lya,,ci 2nd AndynsOK NA NA Coofimrd L%h 2nd AnalynsOK CoafimdHigh 2nd Analyni OK CoofimdHtgh NA 2.._"o>d"A~* s^m._"l,.v-.d...,sO*n.K"."" oca I ;; ow ow ow ow ow ow ow ow ow 2ndAnalynsOK XA NA NA Caofimrd High 2ndAndvaiOK Confimnl L % h Coofimrd L g h I OW :ow Confimrd High CoDfimd High C o n f i m d High CoofimrdHigh Comfimrd High C o n f i m d Hn%h Confimd H i ~ h Coofinxed H-gb HA NA NA NA CLUO N* NA NA 21% 13% 31% I,*% 11% 8% NA NA NA NA N* i*a NA NA NA NA 3M Environmental Laboratory Page 5312001 219 3M Medical Department Study: T6316.1 AhlDT# 092597.1 CovrnceU 6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 D.w Melbod Blt Mamr 811 Mamr 811 w.250 .g/mL C9125OF 3.k Corromlhn .~PFOS qi.L or 96 h Lcq (0 w394 UglrnL) LOQ (0 00394 ug,rnL) Wa (0 WJ94 WmL) L o g (0 w391UglrnL) Lcq( 0 w394 u p i d ) LOQ (0 w394 UgimL) Lcq(" w394 "grid) Lcq(0 MI94 upimL) LOQ(OOa39,upid) Lcq(0 w394 u p i d ) Lcq(0 w394 WbL) Lcq(0 w394 "e',"L) 75% 76% 68% N* NA 78% 97% IWX 112% NA NA NA nolo, 0 0288 00121 LOQ (0 w394 ugrrnL) 0 w70 0 W88 0 n477 0 0325 O"116 OOlW "03"i 00193 0001.8 00393 0 101 0 138 .". 0 0590 " 0% "0788 00251 0 123 0 13, 00301 0 0160 0 0w . 00614 0 0537 0x9 I65 3 03 0 367 I21 2 23 2PI 0674 0 565 2 11 3 53 281 .17 4 93 3 68 2 31 584 666 2 97 6 80 512 RSD Std. Der. MSiMSDW D N* NA NA 6% 4% NA HA 7% 8% HA NA 70 3 ""134 19 9 ""316 60 6 I128 31 9 I63 3M Environmental Laboratory RSD Sed Do. MSlMSD WD NA NA NA 3% 2% NA NA 8% 9% NA NA NA NA NA .L.II.. NA &CQ (0 w977 y/d) 'Lcq (0 w977 u p i d ) 'Lcq(Ow977 W d ) C L c q (0 ow77 "@*) NA C w (0 w977 u p i d ) (Lcq (0 w977 WmL) <LCQ (n w977 u p i d ) (Lcq (0 w977 u p i d ) NA C L c q (0 w977 u p i d ) 32% 5% 40% 5% 38% 28% 20 NA 32% Zn NA 26% In 35% 1% 46% 5% 16% 35% 2n NA 28% 2n NA 37% 2. <LCQ(0 ow77 u p i d ) -LOG (0 w977 upirnL) QoV(Ow977 W d ) .Lcq(ow977 "ghL) 'LOQ(OW977 UyrnL) <LOQ(owI77 UyrnL) .Lcq(ow9?7 "g/"L) rLOQ(Ow977 u p i d ) (LOQ(0 w977 "&L) CLcq (0 w917 UglrnL) CLcq (" w977 "@d) CLcq ( 0 w917 "gl"L) r ~ c (qn w977 u g l ~ ) NA <LOQ(0 w977 uglmq NA CLOQ (0 W917 uglrnL) .LOQ (e w113 W'mL) .Lcq (0 w193 "g/"L) 4 cq (n on493 u y m ~ ) .LZ <G xi:: "8 ,ti, CLcq (0 w493 upirnL) .LOQ (" w493 "&'d) < L c q (0 w493 "gh"L) 0 462 NA NA n 597 om 0 870 0 680 0 322 0 365 0 629 0 582 NA 0 5pv NA 0 375 0 556 RSD Std. k. C.ln.W.I*. 0 376 0 189 0 294 042, I 0 694 E: RSD slb I**. MYMSD RID NA NA NA 9% 11% NA NA 4% 5% NA N.4 78% 63% 2n, N.4 RSD Std. Dm, MSrMSDWD NA NA NA 21% 13% 34% I,% 11% 8% NA NA NA NA ~ NA NA NA NA kfi NA 29 5 NA o 110 NA 30 4 NA 0 I34 NA Page 220 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covancdl6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 Sample D a l l 'EEK 53 RAT SEE G"W Do.= G""p3 Mid h.r 30 0 my*% C91372F C91171F .Y !".rn"J. - - Extnrtb. *....ruS - - \'.L \'wIlw nL I NA P4 I HA I NA I NA I HA I HA I NA I NA I ~ I NA ~ NA I NA I NA I NA I NA I NA I NA I NA - - I NA I NA I N* I NA I NA I NA I NA I NA I NA I NA I NA I NA I NA I NA I NA I NA I ~ I N* ~ NA I NA I NA I NA I NA I NA I NA I NA I NA I NA I NA I NA I NA - - I NA I NA ReCWCV E lrnC*d PF - PFOS Std - C a r m l b n F.UW 0 9275 "02'1 0 9275 09275 09275 0 9275 0 9275 0 9275 0 9275 0 9275 ~ 0 9275 0 9275 0 9275 0 9275 0 9215 0 9275 09215 0 9215 - 0 9275 0 9275 0 9275 0 1275 n 9275 0 9275 0 9275 0 9211 0 9211 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 09275 0 9275 ~ 0 9275 0 9175 0 9275 n 9275 0 9273 09275 0 9275 0 9275 0 9275 0 9275 0 9175 0 9275 0 9275 - 0 9275 0 9275 E d h,ph win win o n w - PFOS C.W. * 208 ' 156 2" 330 26 4 179 14.3 222 17.8 306 24 5 341 27 3 45 3 58 151 12 I 214 172 701 56 2 312 27.4 368 29 5 491 39 3 142 I,, 442 I5 4 475 38 I 363 29 I - 480 38 5 392 314 880 70 5 697 55 8 680 5. 5 837 67 I 717 62 3 613 49 I 751 60 2 464 37 2 640 51 I 761 649 I50 60 I 330 164 274 220 614 49 2 n 911 73 ~ 218 175 917 73 5 274 220 I8k 148 824 66 0 167 131 195 317 195 317 164 132 174 119 154 123 154 284 228 183 - 698 55 9 95 I 76 2 RSD Sld. k. MSlMSD R I D 39 8 7 I8 31 8 I1k 80 I 660 53 6 81 2 SW..,.k VIlW NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA N.4 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA HA NA NA KA NA - PFOSA - Dl1ulbo F.U.. 1 3 I I I I I I - I I I I I I I I I I I I I - I 10 I" I I 10 10 10 10 - PFOSA - Cam. WmL 201 201 338 293 181 211 252 125 - I 80 165 393 268 324 (89 (85 101 279 283 - k15 323 615 275 382 642 305 123 363 2.7 311 270 23 I k74 343 203 - 430 105 120 30s 517 7M 577 112 109 203 637 103 159 224 IC4 - 125 0 305 0 223 0363 I59 224 106 I25 RSD SU. I*". MYMSD RPD 31 6 00751 22 5 00824 18 7 0 118 48 9 0 536 3M Environmental Laboratory Page 5312WI 221 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance# 6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 C9U85.M C9085SM C9085f.M W859M C9U873M C90819M C9Q881M C90896M CPl25JF C91258F C91264F C91269F CP1270F C912l.F C91289F C91294F C91323F C91326F CV133.F C91317F C91352F CPl356F C91359F C91363F C91367F C91371F C91372F .Y U"h0- U"h*W" unlnown UDhWW Uokmow unlmown Uohova limbown Unknown Unknown UOknOW Unknown Unknown Unknown Unknown ~ unknown Unknown Unknown Unknown Unknown Unkmow Unknown Unknown Unknown Unknown SW"l.1. vernkd NA N4 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA keOV0.y coofim mrkd PFOS I I - C.mrrnt"t - d P F O S * W I d .r % .I1 * .323 7.49 6M 3 21 4 I9 621 I7 I35 6 82 ~ 6 78 3M A042100070 3 17 508 A042100071 5 0% 157 AO42IWU7Z 157 a" A012L00076 4 30 310 A042IWO77 3 10 341 A042lw078 3 47 - 4 35 431 227 M82100023 21 7 5 18 591 I5 2 74 5 03 596 4 68 5 32 5 12 4 92 22 0 118 3% 9 9, ~ 8 37 7 U2 20 I 6 47 6 55 6 72 I5 2 21 5 22.3 8 25 8 I8 23 0 19 7 9 18 9 58 ~ UOhOW UllhOW Surmale vermd NA H.6 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA N* NA N* NA NA NA NA NA NA NA NA ElFOSEaH Dlhlk" F..tar I I I I I I I I I I I I I I I I I I I I I I I I I I I I I 1 I I I I I I I I I I 1 - IfFOSU - C.X. Wid om om 0000 om om om om om 0.m am ~ 24 I ow 26 6 ow 22 9 23 7 23.1 24 5 - 22 0 32 8 31 8 27 1 31 2 23 5 210 21 7 29 U 28 I 316 25 6 3 71 146 10 I 7 15 6 1" ~ 18 I 22 7 24 0 I25 20 8 157 28 3 21 6 39 6 I4 4 38, 36 9 10 7 - 156 I5 7 00283 I 0 0216 00144 0018. 0 0369 0 0107 3M EEEnTxaSvl89-57iIronmental Laboratory Page 222 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance# 6329-212 C9OB.IM CPOBSOM C9Q854M CW855M CW856M CW859M CW873M CW879M C90881M CW8%M C9I25JF C9I258F SWW.1. V"W NA HA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA SA NA NA NA NA NA NA NA N* NA NA NA N4 1" NA IW NA 1" NA NA N* NA NA NA NA SA NA NA N4 Pucovrrvronfim ComrYd PFOS WQ (0W492 u d l !a11 -M Y - C.W. 517 356 529 346 .I7 528 357 141 381 324 1n7 261 391 515 .68 19d 321 425 - 430 451 124 761 877 2'91 604 5199 POI 78, 6246 762 3327 I19 164 665 7 116 691 761 198 57, 927 668 147 178 258 708 815 116 251 - 901 '23 1% LACWi colrmmtmt .f#% ",imLor%r(l 5 17 356 5 29 146 4 17 5 28 3 57 2 12 181 3 24 107 2 64 3 91 5 15 168 3% 321 4 25 430 151 22 4 76 88 20 1 6 0 5 80 9.0 79 6 25 76 J 33 I. 9 164 6 46 ~1 1- 6 6 97 '61 19 8 5 74 9 I7 6 .a I47 178 25 8 7 08 8 35 176 25 I 901 7 23 Analytical Report: FACT-TOX-001 LRN-U2103 S".W.,. verw NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA N* NA NA NA NA NA NA NA NA NA HA NA NA NA A : PFOSEA DUutho F.U.I I I I I I I - PFOSEA C.M. llyd. ow ow ow ow ow ow ow ow ow _o_w _ ow ow ow ow ow ow ow ow - ow 17.8 ow ow I1 I OW OW ow OM ow ow 150 ow ow ow ow _0_00 _ ow c 30 ow ow ow ow ow 2 52 ow 0 00 ow 5 60 OW - OM Gw IUD Sld. Dn. W DWD NA NA NA NA NA NA NA NA 3M EEEnTlSv4I-957iLronmental Laboratory Page 223 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance# 6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 Sample Dah EEK 53 RAT SER G"up Drr Crnupl Msd Dox 30 0 mg>kg C91255F C91258P C9126.F C91269F C91270F C91274F C91289F C912PlF C91297F 311 J C91305F CM2M 70 I I C91347F 317 CP1352F 117 C91356F 132 C91163F 123 C91367F 284 CPl37lF CPI372F "1373F 16 2 n.rm& RSD Sld. Do. M-SD RPD 19 8 7 I8 0 489 ".E5 0 .a4 0 179 0 283 33 8 0415 111 (I 323 0.615 I ) 275 0 382 0 662 "0 305 221 0 363 a 247 0331 0 270 0231 0.71 0343 BO I 0 2"1 660 0 430 ~ I as I20 51 6 323 I 6W 321 .I9 I :::621 31.6 00751 -115I , a225 0082, 22 7 5 28 5 91 I52 7, 5 03 596 4 68 5 31 6 72 8.25 23 0 .8 9 0536 , 9 78 1.51 FSD Sed Dn. M W S D RTD 35 9 I72 -LDQ(o w9n " g h L ) rLDQ (0 0248 u d d ) C W (0 0248 u g i d ) 0.0266 C W (0 0248 UgimL) W X (0 0248 u g i d ) .W (0 0248 UglmL) iW (0.0148 UgimL) rW(00148ugimL) 27 0 cLOQ(O.0218U y d ) I I. 0.0328 a0318 0 0274 00312 CLOQ (0 024%udrnL) 'Loo (0 02.8 YgrmL) <LCQ (0 02.8 " g r d ) 0 02m 00281 00316 0 0256 LCQ (a w493 GGL46 00101 69 2 0 w72 6 23 00061 onis1 0 0227 CLOQ(OO118 uymL) 00125 0 0208 00157 0 0283 0 0236 0 03% OOL,. OOlB. (I 0369 0 0.07 51 3 00156 6 5: 00::; SU. Do. M W D RTD u g l d .r % Rn 5 19 346 .17 5 28 3 57 2.42 NA 381 NA 324 4.07 264 3 91 5 45 4 68 396 I 3 21 425 9 76 4 30 0 M252 451 I 224 76 M W S D RPD C.rn"t"U." .~PFOSE* C WWh(0L.0.r49%2 URglnmL) cLOQ(0 0492 u&L) CLDQ (0 Ob91 ILDQ (0 0492 uglmL) cLCQ(0 0492 WmL) 'LDQ(0 0492 u&L) 'LDQ(0 0492 udrnL) <LCQ(O 0492 WmL) 24 3 c m (0 0492 WmLI 400 0970 I I 410 188 0771 RSD Sed Dn. MYMSD RPD NA NA NA NA 462 6 16 52 7 NA 0 w992 116 I04 548 NA 6 97 761 19.8 5 71 9 27 6 1 147 178 25 8 7 08 8 35 176 25 I - 41.7 9 0.1. 55 5 NA 6 w96 I l' ria t.91 NA 3M Environmental Laboratory Page 224 3M Medical Department Study: T6316.1 M D T # 092597.1 Covance# 6329-212 Be. w-009 - EXt..Cti." V.1 mL - I I - I - I I I - I I - I I - I I - I I - I I - I I - I I - I I - I - I I I - I I - I I - I - I I I2~I-RTSMSUL CC1712M - I I I - I I - I CW716M I CC1722M I ~ ~ 7 2 4 ~I CW726M I C911728M I C90735M I C90742M I TW743M I C'X1747M I CW750M I CW758M I CC1765M CW778M C91127F - I I I I I I I I I I I I I - I I I I I I I I I I I I I I I I I I I I I - I I I I I I I I I I I I - I I :n.hulhd S",rn*.k Verlbd NA NA NA NA NA NA NA NA X NA X NA NA NA NA Confimrd Htsh HA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA E NA NA NA NA NA N* NA NA .rru.llgbly.b - PFOS St1 c.r,=,,., F.d.. - n 9275 "9275 - n 9275 0 927, - 0 9275 1) 9275 0 9275 0 9275 0 3215 - 0 9275 n 9275 0 9275 "9275 - "0 9275 9275 1,9275 NA NA NA NA NA NA NA NA - NA NA - NA NA - NA NA - NA NA NA NA NA NA - NA NA "9275 0 9275 0 9275 I,9271 0 9275 "9211 1,9275 0 9215 ,,0 9275 9211 0 9275 II 9275 - ,I 9275 /I 9275 ,I 9175 0 9271 0 9275 11 9275 0 9175 0 027, n 9275 n 9275 n 9275 n 9275 0 9215 - 09275 119275 n 9275 u 9275 0 9171 (I 9275 0 9215 n 9215 "0 9275 9275 09275 "9275 ,I 9275 "9275 "9275 "9275 (I 9275 0 9271 n 9275 0 9275 Q 9275 0 9275 - n 9275 n 9275 0.9275 0 9275 0 9z75 E 0 9275 0 9275 n 9275 n 1275 "U '1275 '1275 - 0 9275 0 9275 hr valid lhne 0 8640 - ,I 861" 0 116," 11.8611 - 0.86," 0 864" 0 8640 ~ NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA - NA NA - 0 86." n 8wn "0 8640 R6411 Analytical Report: FACT-TOX-001 LRN-U2103 RSD wn Sld. Der. MWSD NA NA N* NA NA NA NA NA "% NA 17% NA I% NA 5% NA I% 4% NA 111 1111123 RI 2 0 0237 no I41 111 7 2 65 3M Environmental Laboratory Page 225 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance# 6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 NEEK 105 RAT ! croup Lhlr MeIbod B1k CW724M CW126M C9n728M CW735M C90742M CWlkIM CM717M CW750M CW758M CW765M CWllBM C'11127F C91111F C91138f C9llUF C9ll48F C9111lF C91159F C91166F C91172F C91116F CIII8IF C9118RF C91190F C9n781M CW112M C91235F C91216F C91212F C9IZklF IOlL-,5109 Pms* P",i#y C...~l*o Q..t.r unknown Unknorn Umknowo untrown U"b0W uaknoun unknown UOknOxO Unknown U"ls0W Unknown unknown u...ovn untnown U"kn*W U"k"0wn NA NA N.4 NA NA N* NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA unknown Unknown U"b0Wuo S.",., verw NA NA 2ndAdy$mC NA NA NA NA NA X NA x NA NA NA NA CO.lmrd HIE NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA ti* NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA N.4 NA NA NA NA NA NA NA NA N* NA NA NA NA NA NA E NA NA NA NA NA NA NA NA u & g M y .bow - PFQW Dllulhi F.d. - I I - I I - I I - I I - I I - I - - I I I I - - I I I I - I I I - I I - I I - NA NA - I I - NA NA 1 - 4 I I - NA NA - I I I I 1 I I I I I I I - I I I I I I I I I I I I I - I I I I I I I I I I I I I I I I I I I I - I I I I I E I I I I I I - I I 137% 123% i lZ5% i 143% 1.0% 128% ; 121% i 93% llnn NA NA 91% Ins% NA NA 72% 72% I",% ,"I% NA Rso Std. M . MWMSD RID NA NA NA NA NA NA NA NA JY. 1% 2% 3% 17% NA 12% NA ln% 3% NA NA NA NA 3M Environmental Laboratory Page 226 3M Medical Department Study: T6316.1 ieEK 105 RAT SERA lotT-llll I W O S M QuMy c.rml*n S"rnl.1. V"lM hCl0, Unknown NA Unknown NA Unknown 2nd A d y n a O K Unknown NA Unknown NA Unknown NA Unknown NA Il~knOW NA I Unknown X Unknown NA Unknown X ""bo- NA I Unkaow NA I U"ln0.m NA U"knDWn NA u.know. Con(-d Hssh NA NA NA NA I NA NA NA NA NA NA NA NA I NA NA I NA NA NA NA NA NA NA NA I NA NA I NA NA NA NA NA NA NA NA I NA NA I NA NA NA NA NA NA NA NA 4 NA NA 4 NA NA I NA NA I NA NA I NA NA I U"h0W NA I UOknOW NA I U.k"OW" NA I unlnow. NA I Unknown NA Unknown NA U"knOW7 NA Unknown NA Unknown NA Unknown NA Unknown N.4 ""known NA Unknown NA I Unborn NA llnknown NA Unknown NA Unknown NA llnknovn NA llnkmxio NA U"kn0W" NA Unknown NA ll"k"*- NA Unknown NA Unknown NA Unknown NA Unknown , NA I Unknown ' NA Unknown ~ NA I Unknown NA I Ilnho- NA U.knoun NA Unknown ! NA I U"kn0WI NA In ""known NA 60 Unknown NA Unkmwn NA I Unknown N.4 I Unknown NA in Unknown NA Unknown NA Unknown NA u.k"ov. NA ulllnown NA unknown NA I llnknow~ NA Unknown NA I" Unknown NA Unknown NA Unknown NA Ilnkmwn NA llnkmw. NA Unknown NA 10 Unkmvn E E Unknown NA 1u llnknow NA UolnOWn NA I Unknown NA in UOknOW NA 1" Unknown NA 1" unknown NA Unknown NA knablhmdpakan IiCUylhov. ihr vsh' nc, Analytical Report: FACT-TOX-001 LRN-U2103 R5D Sld. Ik" M W S D RPD NA NA NA NA NA NA NA NA 1% NA 1% NA 8% NA 18% N* 1% 7% 1% NA NA NA NA n611 E I111 llJ811 "609 16" 4 39 2 72 I17 0 8% 62, 0 5.1 9, I I41 3M Environmental Laboratory Page 227 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Sample D m YEEK 105 RATS, c..up w.r MSIhod Blk Croup 1 Low mu 3 "&E S"...l.l. Vlllkd ElFOSWH Di1l bn FU. . NA I NA 2odAomlymOK NA I NA NA NA N* X NA X NA I NA I NA NA Confimrd High NA NA I NA NA NA NA NA I NA I NA NA NA NA NA NA I NA NA I NA NA I NA NA N* NA NA I NA N.4 I NA NA I NA NA NA NA NA NA NA 1 NA NA I NA NA I NA NA I NA NA NA NA NA NA NA NA NA NA NA NA N* NA I NA NA NA NA NA NA NA NA NA NA NA NA I NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA E NA N* NA I NA NA NA NA NA Vly .bow Ihr valid 181 CLGQ (0 011977UgimL) ILcQ(0"2,8"g/rnL) CWQ(""Z.8 "gl"L) cLCQ(fl021R UpimL) C L c q (e 02.8 "glrnL) 3.K 28% NA NA 33% 26% NA NA 2I% 25% 31% 21 NA 21% 25% 38% 2, NA 112% 127% 21% 2% 37% 2n 39% 2n - ~ ~ ~ ( n" Yw~ Lm) 'LGQ i n ow77 "drnL) RSD s*. I*". W S D RPD N* NA NA NA NA NA NA NA 19% NA 23% NA 20% NA 17% NA 4% IIW. 4% NA NA NA NA NA NA NA NA 3M EnETSvI-5irI onmental Laboratory Excel 97 Page 228 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covanceb' 6329-212 Sample D d n EEK 105 RATS G..W Dose Qc-250 ppb Bor IXIMYW L S.mnk# lolNBl1311k7-8 Ms% P'lly Corrrrlbn F.Cl.. 1121171il-H20 Elk unknown vnknorn Unknown Unborn llnhoul" Uohown Unknown Unknown IJnknown ""known Unknown U"knC.wn U"kn0W ""born unknown U"hDWll NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA l1"bOW" unknown 1l"hOW. uaknoua Unknown U"b0,". Unknown lI"h*W Unknorn unknown U"b0.x" U"b0W llnknom U"b0W Unknown ullknown C91166F C91171F C91176F UDknOWa Unknown Unknown Unknorn unborn Uatnom unknown llnhown Unknawo U"b0W" Unboun U"k"0W "OknOVl lUin"kbn*ow- a U.hW UBh0.M unknown Unknown Unkmua ""k"0~ unknown u.tnaw. . Unknown Unknown L l n kn o r Unknow Unknown Unknorn Unknown Unknown Unknorn Unborn llnkmown Unknown untnom Unknown U"hW Unknown lloboro llnbown Unknown Unknown unbowa SW.l.1. Vl.lfkd NA NA Ind A d y m 01 N* NA NA NA NA X NA X NA NA NA NA confirmed H'G NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA N* NA NA NA NA NA NA NA NA NA NA NA NA NA N.4 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA N* NA NA NA NA NA E NA NA NA NA NA NA NA Analytical Report: FACT-TOX-001 LRN-U2103 wn Sld hr. MSlMSD RSD NA NA NA N* N* NA NA NA fI% 4% 1% 1% 1% NA 7% NA 8% I% 3% NA NA NA NA 79 4 ",IS 98 I, (I 667 3M EEnTSvI-iI Ironmental Laboratory Excel 97 Page 229 3M Medical Department Study: T6316.1 AMDTU 092597.1 Covancdt 6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 IEEK I05 RAT SERA ,," m y l g CD(I716M CW726M CW728M Cll735M CW742M I C9075BM CW765M lots29 PFOSEA Purl* Comtbn hCl., Uaknown Unknown Unborn UUbOXn U.k"OW" Umkmvn Unknown U"h*W Unknown Unhown Unhovn URhOWll U0kDOW" Unbow. Ulmwn U"k"0W" NA NA NA NA NA NA NA NA NA NA NA NA NA NA N* NA NA NA NA NA NA SlU.O,.b VWlmd NA NA 2ndAndym OK NA NA NA NA NA X NA X NA NA NA NA Confimd High NA NA NA NA NA NA N.4 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA N.4 NA NA NA E NA NA NA NA NA NA PFOSEA Dllvtro F.Cl.. I NA NA I I NA NA I I I NA I I I NA k 1 I I I I I E I I I I I I "M.""&* LAC0 75% NA NA 79?, 60% N* NA 46% 65% 0% 2 NA 46% 56% 45% 2 NA RJ% 87% 61Y. MI% 57% 2 56% 2 ' W ( I I M l 4 9 2 udmL1 RSD Std. Dw. MYMSD RID NA N* NA NA NA NA NA NA 5% NA 27% NA 34% NA In% NA 4% I% 2% NA NA NA NA NA NA NA NA 3M Environmental Laboratory Page 230 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 ------- ---WEEK 105 RATS G""P D..e Method BIX Mamr BIX M1716M CW~ZM C9072,M CW726M C9n728M CII735M C90741M C94743M C911747M CX1750M CW758M CW765M C9il778M C91127F C91131F C9113RF C9II44F C91148F CIII57F C9II5'IF CVllMF C91172F CP1176F C91183F C9ll88F <-(e I10391 "glrnLi "on "on "on ,,I", 121% 121% NA NA 320% !Ill% NA NA RQ% 83% NA NA 81% 77% NA NA 72% 73% 97% ]Ill% NA N* 'W (0 00394UglrnL) .LW(Oon394 "gld) 001 00116 'w("WIP."glmL) ,10131 1.26 3 69 I89 2 19 0 502 0319 459 n 142 0 104 I1 I75 I1 317 fl86 I 42 4 56 n.87 ,>n 29 184 0215 I59 I SI, 5 89 RSD std. I*". MSMSD WI NA NA NA NA NA NA NA NA Q% NA 17% NA 4% NA 5% NA 1% 1% NA ... . 837% 123% 2 125% 1 143% 1.w 128% 2 131 "0123 87 2 n nz37 120 I .I "QJQ, om43 "0166 n 251 0 309 (I 129 70 7 OM21 2 65 "'.'mgr " I1 0220 113101 RED Sed. Dn. MSMSD W D NA NA NA NA NA NA NA NA 3% 2% 2% 3% 17% NA 12% NA n% J% NA NA NA NA NA 64 n UQll9 1225058 0 1037 NA NA NA NA NA NA NA NA 2% NA 2% NA 8V. NA I "% NA I% 7% 22 NA NA NA NA 62 4 ,! 345 94 I42 3M EEnTSv8.5iIronmental Laboratory Excal 97 Page 231 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance# 6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 Sample Dsls VEEK 105 RAT $ Croup n0.r Mdhod BIk Mamr BIk -LCQ (0 1,248 @"L) -lDQ(OO141ugimL) clGQ(OU148 uglmL) 14% 28% NA NA 33% 26% NA NA 21% 25% 31% 1s NA 11% 15% 18% 2c NA 121% 127% 26% 19% 37Tc la Rso su Dn. MWMSD W D NA NA NA NA N* NA NA NA 1% NA 23% NA 20% NA 17% NA % . Ill% 4% NA NA NA NA I% NA Y1 .6% 7% 56% 45% 1n NA NA RJ% 8% 87% 61*% I% 60% 57% 2n 1% 56% 20 NA 27% 34% NA IRK NA 1% I% 2% NA NA NA NA NA NA NA rLOQ NA NA NA NA NA ""%a LAC"5I": 79. 0115 'JR (I 667 3M EnETSvII-i5 rI onmental Laboratory LlUl91 NA NA NA NA Page 232 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance# 6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 Study P d w l NumheOesf Substance) Malnr 104 Week DietaryCarsmogenicity Study with N a m w Range (98 1%) NZthyl Portlua~tancsulfonamimidoEthanol in Rats T-6316 (EIFOSE-OH) Metho4AYRevision Analytical Equipment SystemN u m k Instrument SoRwa~Ncrainn Rlenam R-SquaredValve Slnp Y-Intercept Dalca of Exlraclindhalysl Dates ofAnalysia'Analyal Date ofData R e d u l i u d h l y a t ample Data VEEK 105 RAT SE Gmvp Dose Smnplc # Gmup 3 Mid h u 30 I1 mgkg C90842M C'XIU44M CW8511M C'Xl85IM CH1854M C'Xl85SM CW856M CWRSSM C9088lM C908MM CW865M C90867M C90869M C'21871M C90881M C90882M CW883M C90884M C911885M CSilRR9M C'Xl893M C909UOM C91252F C91253F C91254F C91256F C91264F C91278F C91279F C91286F C91292F C91294F C91295F C91301F C91303F C913U7F C91310F ,,e - Etrnclioo - Vol. mL I I I I 1 I I I I 1 I I I I I I I I I I - 1 I I I I 1 1 I I I I I I I I - 1 1 S"rr0p.e vermed NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA - F'FOS Sld - Comr11on Fartor 0 9275 n 9275 0 9275 I! 9275 t l 9275 tl 9275 n 9275 11 9275 tl 9275 n 927s 11 9275 11 9275 I1 9275 0 9275 0 9275 I1 9275 0 9275 t l 9275 n 9275 I! 9275 - n 9275 n 9275 n 9275 n 9275 n 9275 I1 9275 n 9275 n 927s I 1 9275 n 9275 I1 9275 I! 9275 t l 9275 11 9275 11 9275 - I1 9275 0 9275 - C0"C. Og/d 1u2 115 273 224 683 483 I12 670 2nu 667 4n2 122 I55 420 4NI 592 374 391 623 I59 -578 387 577 674 I19 823 571 138 139 868 210 774 185 753 128 - 883 665 Coocmlr.tlon Of PFOS "g/rnL 0, Y. Rcc 40 7 4 61 I 1 11 8 98 27 3 I9 4 4 49 26 9 83 2 26 7 16 I 48 9 6 22 168 I84 23 7 IS0 157 24 9 6 35 23 2 I5 5 46 2 54 I1 95 5 frJ n 45 7 I10 I12 69 5 16 9 62 n 148 , 611 4 51 4 70 8 53.3 "arm& rooamidoacetate M S E = N m w Range N-EIhyl Perfluomoelmrsulfonmimidaethyl alcahol k1556 = C8Fl7S02N(Rl)cHzl:CO) PFOSEA = Perfuamtane aulbnyl rlhylamidc RSD Sfd. Drr. MSrmSD RPD 80 0 17 6 49 7 3M Environmental ETS-8-5 I Excel 97 Laboratory Sera Week IO5 (2) TOXMI-ura-2I2-ItAI XIS Page 233 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 WEEK 105 RAT SE L GlOVP Dole Sample U Group 3 CW84M NA Mid Dose CW844M NA 311 I1 mgn* CW85OM NA CW85IM AA CW854M EA CW855M C911856M h'A C90858M NA C908N1M NA CIIIMM NA C9086SM NA CWI867M NA CW869M NA CW871M NA CW881M NA CW88M NA CW883M NA C90884M NA CW885M NA CW889M NA CW893M NA CW'XW)M NA Group 3 C91252F NA Mid Dose C91253F NA 30 n C91254F NA C9I256T NA C91264F NA ~91278~ C91279F NA CY1286F NA C91292F NA C91294F NA C9129SF NA C91101F NA C913113F NA C91107F NA C91310F PFOS = PernuomDetanegY I k PFOSA = Perfluomtsnesulfonamide PFOSAA = PerfluormdanesulfodamidoaEetate EIFOSE = N m w Range N-Ethyl PerfluomlsneaulfonaM*,ethyl almohol MSS6 = CSF17SO2N((H)CH2C03) PFOSEA = Prrfuomoslane sulfonyl ethylamids - PFOSA - Dilution P.CtW I I I I I IO I I I I I I I I I I I I I I - 1 I I I I 1 I IO I I I I I 1 I - I I - PFOSl -Con& OghL I96 15s 187 81 7 287 14 6 223 217 138 272 238 4311 258 343 S36 482 436 232 414 315 -459 296 369 484 380 572 3411 71 7 206 497 447 542 306 263 368 -S36 4111 -- CO.CLOtI.tII,. -- d P F O S A vymL or K R r 0 196 0 155 I1 187 11 0837 I1 287 11 346 11 223 I 1 217 I1 338 0 272 I1 238 I1 4311 11 258 n 143 I1 536 ii 482 I1 436 0 232 0414 11315 --0.45Y I1 296 11 169 11 484 0 380 I1 572 I1 3411 0717 I1 206 11 497 11 447 11 542 I 1 3116 I 1 263 11 368 I1 S I 6 RSD std. Dcv. MSlMSD RPD 37 9 11 116 30 9 I1 113 3M EEnTSv-8-i5rI onmental Laboratory Excel 97 Sera Weck 105 ( 2 ) TOXoOl -sera-ZI 2-1 I AI XIS Page 234 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance# 6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 Sludy Roduet Numbeflest Substance) 104 Weak Dietary Carolnogendiy S M y wlh N m w Raoge (98 1%) N-Ethyl Rrflvomoctanesulfm~do Ethanol UI Rats T-6316(EtFOSE-OHI Sample Data VEEK 105 RAT SE L Group Dow s.mp1. x crovp 1 C90X42M Mid Dose C90844M 30 11 C9085OM C9085IM CW854M CW855M CW856M C90858M C90860M CX1864M C10865M C90X67M CW869M C90871M C908XIM C90882M C90883M CW884M C'Xl88SM C911889M C'X1893M C90'XiIM crovp 1 C9125ZF Mid DDse C91253F 30 I1 m&g C91254F C91256F C91264F C91278F C91279F C91286F C91292F C91294F C91295F C91301F C91303F C91307F C91310F 'OS = Pernvomrlanesul le 'OSA = Perfluomoolanes OSA,=Pern"omrlan< FOSE = N m w Rangeb 556 = CHF17SOZN((H)C OSFA = Perfvomoofanesulfonyl efbylarmde SWr0g.f. VWiIid C o n l i i Rgh NA NA NA Nh NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA - PFOSAA - Dilution P.tt*. IO 111 IO IO IO 10 IO Ill 10 IO IO 10 IO in IO IO IO IO IO IO - Ill IO IO IO 111 IO in iim 111 IO in IO IO IO in - 1u IO - C0.C. "@lL 187 132 308 98 4 715 128 509 379 376 355 166 287 455 3311 472 3911 435 2% BIS 652 - 481 647 373 733 646 791 316 95 9 334 932 525 781 614 349 524 - 721 614 -- CoOnotl.lioo -- ofPFOSA4 udmL or Rec 1187 1 :I2 308 n 984 7 IS 8 :!8 5 119 3 79 3 76 3 fS 1(16 287 4 f5 3 ?ll 4 il 3 $0 4?5 2 56 6 1.5 6 52 --- 4 XI 40 4 6 47 4 I4 I92 173 7 33 6 16 7 91 3 16 9 59 3 34 9 32 5 25 7 81 6 I4 3 49 --- 5 24 7 21 614 6 14 34 2 2 In 3M Environmental ETS-8-5 I Excel 97 Laboratory Sera Week 105 ( 2 ) TOXOOl-sera-212-1 I AI XIS Page 235 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covancdl6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 Sample Data VEEK 105 RAT SI Group Do.* croup 3 Mid hss 311 I1 mgiLg CW856M C'Xl858M C'XI8lXlM CH)864M CH1865M C'X186lM CW869M C'X1811M C'Xl88IM CW882M C'XM83M C'X1884M CW885M CHl889M C'Xl893M CXI'XxIM C91252F C91253F C91254F C91256F C91264F C91218F C91279F C91286F C91292F C9I294F C91295F C913OIF C91303F C913lllF C9I3IUF 104 Week Dietary Carcinogenicily S M y with N m w Range (98 I%) N-Ethyl Perfluom:tancsulfoddo Ethanol in R:tls T-6316 (ElFOSE-OH) Rat Sem ETS-84 I & ETS-8-5 1 h l i s 1162498,Davey 010799 M a d p 3 3 8134 See Below Sec Atlachments See Attaehmenls See Attacbmonts I I ~ ~ I I XILIIO.~IHI. UZJO~IIXI. m411ii RWW, SAL 0zi18/W, 0U25lOll.03/211W,00111lW, 0019/~10.06120/00,118/1lUlXI. 0 6 1 I i ~ W0.8123lM1 MMHnAS ll2L?I/lM~, 112I29/W,02/29/W, Uidl16nX1, 06/llMxI. ll02lllW, 06/26/IXl, 081l13/1)0, I I R I l 81W IllllZSllXl MMHIIASNOJ UnknOUm UoknOUn UhOWn UnLnom LlnLnOUn UnLnOUn SWO*.k VIIIlid NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA 2nd Analysis OK NA NA EIFOSE-OH Dilution Faelor I I I I I I I I I I --E1F0SE43A --C0"C. "gtoll s IS I89 5 IO 3 82 I 'XI 3 37 3 16 3 30 490 10 3 5 41 6 66 6 06 2 MI 9 89 I8 2 I86 3 89 I5 2 171 --IO 6 4 68 8 42 8 56 21 3 564 5 11 17 n I 'XI 6 46 I5 3 128 n sin 6 03 9 21 --1 2 8 I36 Pllmsmc AV.r.gC E1FOSE-OH VglolL RSD Std. De?. MSlMSD W D fl(HI9IZ I 1 11108 3M EEEnxTcSev-l89-i57rI onmental Laboratory S o n Week 105 (2) TOXMXII-~n-2l2-IIAIXIS Page 236 3M Medical Department Study: T6316.1 study Ralucl NumkqTesl Substance) Matrix Melhod/Rewsmn A~~alpicaEqluuulpmeol SystemNumkr lnsmunenl SoRwamNemon Filename R - s q d Valve slop. Y-lntereept Dales of Exlracliodhlysf Dales ofAnalyadhalysl Dale of Data ReducliodAnalyst ample Data IEEK 105 RAT SE GlOVP Sample Y os = Perllwmotanesvl OSA = Perfluomlanes OSAA = Pernuomoelanc CO842M C90844M CHl85OM CO85IM CHl854M CHl855M CW1856M CWI858M CHI8MM CO8MM CStt86JM CO867M CO869M CO871M CHl881M C90882M C90883M CStl884M C11885M ~~1889~ CO893M CStIStXIM C9lZJZF C912J3F C912J4F C91256F C91264F C91278F C91279F C9128W C91292F C91294F C9129JF C91301F C91303F C91307F C91310F le nmde boa mdasceIaIe co,,~llao Psclor UnknOW unknown UnknOUn AMDT# 092597.1 Covsnce# 6329-212 ConfimdHieh NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA M5M Dhlion P.clOl IO IO in in IO IO in in in in IO IO 1 I1 in IO IO In IO IO IO IO IO IO Ill 10 in IO IO IO 10 Ill 10 IO IO IO in in --M556 --Cone. ndml 254 116" 214 I69 187 162 270 610 2c-l 228 337 zn8 270 I92 378 294 J34 334 284 4411 --252 286 385 474 324 291 178 223 176 312 470 346 209 348 381 --12s 1x1 Analytical Report: FACT-TOX-001 LRN-U2103 RSD Sld. De". 1.69 187 I 62 2 7X 6 IO 264 2 28 3 37 :on 2 70 192 3 78 2 94 5 34 3 34 281 4 40 41 7 1191 2 00 3 40 3 81 35 7 I 118 3M EEEnXT-Sv1-987i-JrIonmental Laboratory Sera Week IOJ (2) TOXOOl-~m-212-1IAI XIS Page 5131Q.lKl1 237 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covnnce# 6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 ample Data YEEK 105 RAT SE Gmvp Do* BOX in14419 ID, s29 Correctlo" Group 3 C90842M NA ' I Mid Dose CIXl844M NA I 30 t i mgng C9085UM NA I C90851M NA I CY1854M NA I C908SSM NA I CY1856M NA I CYI8S8M NA I CY18MM NA I C90864M NA 1 C9086SM NA I C90867M NA I C90869M NA I C90871M NA I CM881M NA 1 C90882M NA I C90883M NA I C90884M NA I C9088SM NA I NA I NA I NA N1 Group 3 Mid DDse 3 0 n m&g C91253F C912S4F C91256F C912MF C91278F C91279F C91286F NA C91292F NA C912!XF NA C91295P NA C91301 F NA C91303F !nd Analysis OK C91307F NA NA C91310F NA NA Ite PFOSA = Perfluomostenesulfo-amidp PFOSAA = P e r f l u o m o o t a o e s v l f o n a ~ ~ ~ l = l ~ EtFOSE = N a m w Rmge NEIhyl Perlluomoetanrsulfonarmdo ethyl alcohol MSS6 = CXFI7SO2K((H)CH2CCO) PFOSEA = Prrfuonwrtaoe sulfonyl elhylamide uste EnlerediBy ~~t~ veniieuB~ Purity EnIerelWenfled I13!116/W, 031101100. 0312il~IXI0. 3/21/W. 0411M I O . 05116/tnI, I16IIMX1. 06/2YIXI,116123/W, tl6l261oO. lI8ilS~lHIl,l9ltlSlW, 111117llXl LAG 'ICSH ~i3111iihi nj i n413nini LAC 01/191i11 LAC 3M EnEETxvcSe-i8l9-rS7oI nmental Laboratory sera Week 10s (2) TOXW-sera-212-llAI XIS Page 238 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Sample Data iEEK 105 RAT SE Gm"p Dow Sample Y Grovp 3 Mid Dops 30 0m&g Gmup 3 Mid Dnne 30 11 m@kg 3s =Perfl"oml?Jlesull PFOSA = Perfluomoctanest PFOSAA = PernuomoctEtFOSE = Narmw Range N M556 =CSFI7SOZN((H)CI PFOSEA = Perrmuama'taneI CW842M C90844M CwnsnM C90851M C90854M C!XIJIM C90856M C'X)858M C908MM C908MM C90865M cm6m l390869M CRl871M C90881M CW882M C90883M CW0884M CW88SM C90R89M l'R1893M ('90XX)M C91252F C91253F C91254F 1391256T 17912648 13912781 1-91 27PF ll91286F C91292F C91294F C91295F C91301F C91303F C91307F 1'91310F le LaMde Conee"tr.floo Of PFOS "dmL or x R4c 40 7 4 61 11 I1 8 98 27 3 194 4 49 26 9 83 2 26 7 16 1 48 9 6 22 168 18 4 23 7 IS0 IS 7 24 9 6 35 23 2 15.5 46 2 54 0 95 5 66 11 45 7 1IO 112 69 5 I69 62 0 14 8 60 4 51 4 70 8 53 3 RSD Std. I k v . MSlMSD RID XI) n 176 49 7 27 4 11 3459MNl n 223 11217 n 338 n 272 I1 238 n 4311 n 258 0 343 0 536 I1 482 I1 436 I1 232 0414 11315 11 459 tt.2M 0 369 n 484 0 380 I1 572 n 340 i t 71731~x1 0 206 0 497 I1 447 0 542 I1 3116 n 263 I1 368 0 536 n 4111 37 9 ,1111 - 3u.9 I1 133 CO.C.Ofr.LIO. "CPFQSAA . VymL or Y. 187 Rs I32 3 08 I1 984 7 I5 8 28 5 09 3 79 3 76 3 55 I66 2 87 4 55 3 30 4 72 3 XI 4 35 296 6 05 6 52 4 81 6 47 3 73 7 33 6 46 7 91 3 16 9 59 3 34 9 32 5 2s 7 81 6 I4 3 49 5 24 7 21 6 14 RSD SId. I k v . MSlMSD RPD - 46 4 4 14 - 6 14 34 2 3M Environmental ETS-R-5 I bXcel97 Laboratory sen Week IO5 (2) TOXml-sera-212-1IAI XIS Page 239 513112001 3M Medical Department Study: T6316.1 study Pmducl Numbor(Teesl Substance) Matrix Malhod/Reviaion. Amlflieal Equipment SystemNumber Inalrument SafiwanNeraion Filename R-SquaredValue slope Y-lnt~rccpl Dales oPExtracfiodAmalyst Dates nfi\nalysiaihalyst Dote ofDafa ReduFtiodi\nalyat Sample Data L4 S.rnr,l. # CHI84ZM CW844M CWRSOM C'XIBSIM C90854M C90855M C!X)856M CW858M cxinmM C'X8MM C4865M CW867M C90869M C90871M C90881M cxianm C90883M CP0884M CP0885M C4889M C!X1893M C'XMNM CPIZSZF C9I253F C91254F C91256F C912MF C91278F C91279F C91286F C9129ZF C91294F C91295F C91301F C91303F C91307F C9131OF - 47 2 11 00430 Con.mlr.tion ofM556 uglmL or % Rrc 2 s4 11NI 2 I4 Id`) I x7 I d? 2 78 6 IO 264 2 28 3 37 2 08 2 70 I 92 3 78 294 5 34 3 34 2 84 4 40 2 52 2 86 3 85 4 74 3 24 2 91 I78 3 23 I76 3 I2 4 70 3 46 2 09 3 48 3 81 I25 I81 Analytical Report: FACT-TOX-001 LRN-U2103 RSD SLd. D w . MSJMSD RPD NA NA NA N* 3M EnEExTvcSe-l8i9-r57Ionmental Laboratory Page 240 3M Medical Department Study: T6316.1 AMDW 092597.1 C o v a n d 6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 Study. F'rcduct Number(Test Substance) 104 Week Dietary Carcinogenicity Sludy wilh Narrow Range (98.1%) N-Ethyl PerfluomwtanesulIooaido Elhanol in Rats T-6316 (EIFOSE-OH) MdUiX. Rat S a m MelhodiRevision: ETS-8-4 1 & ETS-8-5 I Analytical Equipment System Number lnmment SoftwareNmian: Soup 020199, Davey 070799 Masslpx 3 3 & 3 4 Filename. Sce Below R-Squared Value See Atla~hments Slop%: See Attachments Y-lnterccpt See Attachments Dales of ExUaclionlAnalyn: 02/07/W, 02108lW. 02/09/00 RWW. SAL Dales or Annalysidhdysl 0U18100,U28/00. 03127/00.05/31/W, 06119l00. 06120/00,08/03/W. 08/16/00, 081231CO MMHAAS Dale of Data Reductiodhilyst: 02/21100,02/291~,31021000.6102100, WZWW. 06/21/00,08/07100, 08/18/00. 08/25/(0 MMHAASIHOJ Sample Data 'EEK 105 RAT SE GlOOP Dole Group 4 Mid-High Dose Sample # C909MM C90908M C90910M C90911M C90916M C90926M C90927M C90928M Box 00.009 Extr~ctl~n VOI. mL I 1 I I I I I I Sorrobltr Verified NA NA NA NA NA NA NA NA - PFOS Sld - Corredioi F*dW 0 9275 0.9275 0 9275 0.9275 0 9275 0 9275 0.9275 0.9275 l M 171 PFOS Purity Correction Fador 0.8640 0.8640 0 8640 0 8640 0 8640 0 8640 0 8640 0 8640 - PFOS - Dilution Fador 200 200 50 200 200 200 50 200 --PFOS --C0"C rig/mL 417 443 786 91 I 551 384 548 791 W62000016 DO62000017 A053100106 DO62000018 W62WW19 Do62woO20 A053100108 Do62wW23 ('onerntrflo. of PFOS IIg/rnL or % ReC 66 9 71 0 31 5 146 88.4 61 6 22 0 127 C90933M 1 NA 0.9275 0 8640 200 286 W62wW24 45.8 C90944M I NA 0 9275 0 8640 200 848 DO62000025 136 Group 4 Mid-High Dose Immg/lrg C90947M 1 C90948M I C90953M I C90955M I C90956M I CW965M I CW968M I ~91315~ I C91324F I C91327F I C91332F I C91335F I NA 0.9275 0 8640 200 899 Do62000026 144 NA 0 9275 0 8640 200 323 DO62000027 51 7 NA 0 9275 0 8640 50 324 A053100113 13 0 NA 0 9275 0 8640 200 400 DO62000030 641 NA 0 9275 0 8640 200 822 DO62000031 132 - - -- NA 0.9275 0 8640 200 857 137 NA 0 9275 0 8640 200 626 I00 NA 0 9275 0 8640 200 762 122 NA 0 9275 0 8640 200 829 133 NA 0.9275 0 8640 20 612 9.81 NA 0.9275 0 8640 504 627 W81600030 251 NA 0 9275 0 8640 200 62 1 DM2000040 99 5 C9 I336F I NA 0 9275 0 8640 500 504 DO81600031 202 C91338F I NA 0 9275 0 8640 200 875 Do62000044 140 C91340F C91341F C91342F C91356F C91357F C913WF I NA 0.9275 0 8640 500 628 W8IM10032 252 I NA 0 9275 0 8640 200 474 D062WW46 76 0 1 NA 0 9275 0 8640 200 761 DO62000047 122 I NA 0.9275 0 8640 20 386 DM2000048 6.18 I NA 0 9275 0 8640 500 500 D081MXK33 200 I NA 0 9275 0 8640 200 881 W62woOS2 141 C91361F 1 NA 0.9275 0 8640 200 815 DO62000053 131 C91362F I NA 0 9275 0 8640 200 I58 DO62000054 25.3 C91372F I NA 0.9275 0 8640 200 367 Do62000055 58.8 Groop 6 Mid-High Dose C91376F C91378F C91380F C91047M C91054M I NA 0 9275 0 8640 200 83 I DO62wW58 133 - - -- I NA 0 9275 0 8640 200 682 DO62000059 109 1 NA 0 9275 0 8640 200 648 Do62woO60 IM 1 NA 0 9275 0 8640 1 231 A053 I00082 0 I85 I NA 0.9275 0 8640 1 !7.7 A053100072 0 0222 ReC0"ery 100 mgikg C91056M I C9IOM)M I C91065M I NA 0 9275 0 8640 5 293 A053100060 I18 NA 0.9275 0 8640 I 177 A053100074 0 142 NA 0.9275 0 8640 I 0.00 A05310W75 LOU (0 0198 ugimL) Group 6 Mid-High Dose C9107OM C91071M C91072M C91073M C91076M C91451F C91453F 1 NA 0 9275 0 8640 1 li4 1 A053100078 00514 I NA 0.9275 0 8640 1 .I80 A053100079 :LOU (0.0198 ug/mL) I NA 0 9275 0 8640 I i6 6 A053100080 0.0454 - - -- I NA 0.9275 0 8640 I ll.00 A05310008I :LOO (0.0198 ugimL] I NA 0 9275 0 8640 I 739 A053100085 0.592 I NA 0 9275 0 8640 10 126 A053100040 5.02 1 NA 0 9275 0 8640 10 !IO A053100043 1.68 RCO"ny 100 mgng C91455F I NA 0 9275 0 8640 10 739 A0531000M 5 92 C91459F I NA 0.9275 0 8640 I 0.410 A053100089 LOQ(OO198ugimL) C91462F I NA 0.9275 0 8640 IO i76 A053100046 4 61 C91467F I NA 0.9275 0 8640 I :Is 8 A053100093 0 0207 C91473F I NA 0 9275 0 8.540 10 ,479 A0531wO50 3 84 C91479F I NA 0.9275 0 8640 I I1.W A053100095 LOQ (0 0198 ugimL) C91480F I C91485F I C91487F I C91488F I - 'OS = Perlluorwctanesu late PFOSA = Pcrlluomoctlnesulfonamidc PFOSAA PerlluarooctancsulIonaidolcefate EtFOSE = Ndmw Range N-Elhyl Pemvorwctanesulfonamido NA NA NA NA ~rrectedPFOS -new LOQ is 0 01 A data is above elhvl alcohol 0 9275 0 9275 - 0 9275 0.9275 ' (0.0247 ug glmL LAC ipper limit, 0 8640 I 09 0 0 8640 10 ,443 o 8Mn 0 8640 ) t o include std 19101 - -- I I. 20 10 i19 antitation and is provided as an cslimate A0531W096 A0531XWJ 1 A053100101 A0531XW52 0 0553 3 55 LOQ (0 0198 ugimL) 4 I6 M556 = C ~ F I ~ S O ~ N ( ( H ) C H ~ C O O ) PFOSEA = Pcrfuorowtaiie sulfonyl cthylamide - Average - PFOS og/mL Sld. Dav. MSlMSD RPD - 84 6 53 6 45.4 - 122 57 9 70.5 - 0 227 I66 0 377 - 2 41 96 3 Dale EntercdiByDate Verified/ By. purity EnteredIVcrifid. 03/06/00.03/10lW, 03120100, 03/211W. 4/15/00,5116100. 06/13/00, 6/15/00. 6/19/00, 06/22100.06123/00.06126/1!0. 08/15/00. 09105100, 10117/00 LAUCSH 04l30101 hoj /04/30101 LAC 02119/01 LAC 3M, EEnETxcSve-l8i9-r57 Ionmental Laboratory Sera Week 105 (3) TOX-001-sera-212-1I A IXIS Page 241 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covin& 6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 Study. 104 Week Dielaiy clninogenicity Study With Narmw Range (98.1%) N-EUiyl Prrfluomlanesulfonamida Ethanol in Rats hoducl N u m W T c s I Subnmce). T-6316 (ElFOSE-OH) Mhi- Rat Serum MethodiRevision. ETS-84.1 Bi ETS-8-5 I Andpical Equipmen1System Number: So"p020199.Da"ey070799 InsUUment SoRwareNersion: Maslynx 3.3 Bi 3.4 Filename. see Below R-Squared Value. See Allachments Slope. See AItachmmts Y-lnlenepc Dales or ExUactiodAnalyn See Auaehments W07100, 02/08/00, 02109100 RWW, SAL Dates of Analysislhalyst 02/18/00. 2/28/00, 03/27/00, 05/31/00,06119/00.0612WW. 08/03/00, 08116/(Q 08/23/00 MMWIAS Dale of Data ReductioniAnalyst. 02/21/00, 02/29/00, 3/02/00. WOUW. 06120lW. 061ZIl00,08/07/00, 08/18/(10. 08/25/00 MMWIASNOI Sample Data YEEK 105 RAT SE L GlOllp DOSL crwp4 Simple I c90904M Box M)-oo9 Iot L-15709 PFOSA Purity Correction Factor mknm sorrogrtr Vrrlflrd NA - PFOS/ - Diludo Facto, IO - -- PFOSA Filename - -- cone. nymL 56.8 P.0227W090 -- ConcrnlrPllo" -- ofPFl3SA m%/RLor % Rec 0.5118 Mid-High Dose C90908M U"kn0W NA 10 60 6 P 022700091 0 6116 IWm%Lg C90910M C90911M U"lm0wn U"kWwn NA 10 I84 P 022700094 I84 NA 10 60.7 PO22700054 0 607 C90916M U"kn0WI NA IO 61 2 PO22700055 0.612 C90926M U"kn0W NA IO 33 0 PO22700095 0.310 C90927M U"kn0WlI NA 10 109 A022700096 109 C90928M unknown NA 10 97 9 A0227W097 0 979 C90933M U"kn0WI NA 10 59 3 A022700698 0 55'3 C90944M U"hl0WI NA 10 56 0 A022700056 0.5tQ CW947M Unknown NA 10 68.5 A022700059 0.695 C90948M unknown NA 10 27.7 A022700060 0.2i7 C90953M Unknown NA 10 57.2 A022700IOI 0 572 C90955M U"kn0wn NA 10 61 4 A022700102 0.614 CW956M U"kn0W NA 10 39.0 AO227W061 0.390 croup 4 C90965M C90968M C91315F U"kn0Wn U"lm0WlI Utllinown - - -- -- NA 10 94.9 A0227W062 0 949 NA 10 63.8 A0227W103 0 638 NA 10 89 8 A022700077 0 898 0.701 Mid-High Dose 100 m& C91324F C91327F U"h0Wn UnlOlOWlI NA 10 191 A022700080 I91 NA 10 I02 A0227W081 1 o:! C91332F C91335F Unlmown UilknOW NA 10 136 A022700082 136 NA 10 205 A022700083 C91336F U"kn0wn NA 10 107 A022700084 C91338F U"kn0W NA 10 171 A322700087 1.71 C91340F unknown NA 10 72 I A322700088 0 721 C91341F Unknown NA 10 138 A322700089 I3LI C91342F U"h0wn NA 10 82.6 A1227W063 0.826 C91356F U"kn0W NA 10 34 5 A)2270W66 0.345 C91357F U"kn0Wn NA 10 201 Al227W067 2.01 C91360F U"h0Wn NA 10 185 A)22700068 I8f C91361F U"h0W NA 10 116 A'1227W069 LIf1 C91362F U"kn0W NA 10 72 0 A'l22700070 0.721) C91372F U"kn0wn NA 10 161 A'122700073 1.61 C91376F Unknown NA 10 193 A'122700074 1.91 C91378F C9138OF C91047M C91054M C91056M U"kn0Wn U"lm0W U"kn0W U"lrn0Wn UilhOWll - - -- -- NA 10 117 A0227W075 l.li NA 10 89 I All22700076 0.89L I30 NA I 0 00 A062000022 :LOQ (0 OW23 ug/mL) NA I 000 Al1620MM23 :LOQ (0.W23 ugimL) NA I 000 AM20W026 :LOQ (0.03423 uglmL) C9 I MOM C91065M U"kn0wn U"known NA 1 :LOQ (0w 2 3 UgimL) NA I 000 A142MM028 :LOQ (0.004'23 ugimL) C91070M U"h0UlI NA I 000 Al162woO29 'LOQ (0 004'23 UgimL) C91071M Unknown NA I 000 A(162W0030 LOQ (0 004'23 UgImL) Group 6 Mid-High Dose C91072M C91073M C91076M C91451F C91453F unknown UllhroWn - -- NA I 000 A1162000033 LOQ (0 004'23 ugImL) NA 1 000 AI62000034 .LOQ (0 w493 UgImL) NA 1 .LOQ (0 00493 ugimL) <LOQ NA I 'LOQ (0 004,)3 ugimL) NA I 0.00 A062W0037 LOQ (0 00493 ugimL) Rnovery I 0 0 mgikg C91455F C91459F NA I 0.00 A(1620W0 LOO (0004'13 u g h L ) NA I 0.00 A ( 6 2 0 W 1 LOQ (0.004'13 ug'mL) C91462F NA 1 0.00 Ai62W0042 LOQ (0 004!13 u&L) C91467F C91473F C91479F NA I 000 A(162000043 LOQ (0 004!13 u&L) NA I 000 A(62W0044 L W ( O . W B 3 Ug'mL) NA I 0.00 AC62ww47 LOQ (0.004!13 ug.'mL) C91480F C91485F NA I 0.M) A(162000048 'LOQ (0.004!13 ug'mL) NA I OW Ai62000049 LOQ (0 W 9 3 um'mL) C91487F NA I P9911RXF NA I inected PFOS LOO, 247ugi PFOSA = Pemuorooctancsulfonaide PFOSAA = Pcrfluomoctanerulfonaidoacctnte -n w LOQ is 0 0198 ug. L~ AC . A data is above the upper limit o EIFOSE =Narrow Range N-Ethyl Pcrflnarwctanesulfonaido ethyl alcohol -- LOQ (O.W!D uy'mL) LOQ (0.00493 uy'mL) Is 9/01 lntitalion and is provided as ao cslimale <LOP M156 = CSF17S02N((H)CHZC00) PFOSEA = Perfuomctanc sulfonyl ethylamide RSD Sld. De". MSrmSD RPD 51 9 0 363 39.5 0.512 NA NA NA WA Dale EnlerdBy Dale Verified/ By: Punty EnteredNerified: 03/06/00,03110100,03/20/00,03/21/00,4/15/00,5116100, 06113/00,6/15100,6/19lW,O6/22/00,06123/H, 06126100, 08/I5/00, 09/05/00, 10/17/00 LAWCSH 04130101 hoj /04/30/01 LAC 02/19/01 LAC 3M EnEExTcvSe-l8i9-r57 Ionmental Laboratory Scra Week 105 (3) TOX-001-sera-212-1IAI XIS Page 242 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covnnce# 6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 Study Roduet Number(Tea Subnmcc) Matrix MelhodiRevisioa: Analylical Equipment System Number lnsmmenl SoftwareNenion Filename- 104 Week Dietary Carcinogenicily Study with Narrow Range (98 1%)N-Ethyl Pe~uorwetanesulfonamidcEthanol in Rats T-6316 (EtFOSE-OH) Rat Serum ETS-8-4 I & ETS-8-5 I Soup 020199. Dwey 070799 Masslynx 3 3 & 3 4 See Below R-Squared Value. slopeY-Intercept' Dares of ExVaeIiodAnalyst Dale of AnalysisiAnalysl: See Attachmenls See Attachmenls See Anachmene 02107100, 02lO8100. 02/09/00 RWW, SAL 02/18/00, 2128100, 03127100, 05131100,06119100. 061201W. 08103100, 08116100, 38123100 MMWlAS Dale ofData ReductiodAnalyrt: 02121100,02129100,3102100, 06/02/00. 06120100,06121100.08107100, 08118100, 18125100 MMWIASIHOI Sample Data BOX 00-009 WEEK105 RATSE IolT-7121 I ClOOP Sample # PFOSAA Pmrity surrope Dose Correctloll Verified FIdOl croup 4 W04M Unknown 2nd Analysis OK Mid-High Dose C90908M Unknown NA I00 mdkg CSfI910M U"kn0Wn NA C90911M U"kn0WI NA W16M U&OW 2nd Analysis OK C90926M U"kn0Wn NA C90927M U"kn0Wn NA C90928M U"kn0wn NA C90933M Unknown NA W44M unknown NA W47M Unknown 2nd Analysis OK C90948M UllknOUm 2nd Andlysis OK W53M U"kn0W NA C90955M U"kn0W NA C90956M C90965M C90968M Unknown U"kn0W 2nd Analysis OK 2nd Analysis OK NA Group 4 C91315F NA Mid-High Dose C91324P NA 100 mS"kK C91327F NA C91332F NA C91335F NA C91336F NA C91338F NA C9134OF NA C91341P NA C91342P NA C91356F NA C91357F NA C91360F NA C91361F NA C91362P NA C91372F NA C91376F NA C91378F NA C91380F NA Croop 6 C91M7M NA Mid-High Dose C91054M NA Recovery C91056M NA 100 m a g C910M)M NA C91065M NA C91070M NA C91071M NA C91072M NA C91073M NA C91076M NA Croup 6 C91451F NA Mid-High Dose C91453F Unknown NA ReCOVOy C91455F Unknown NA 100 mg/Lg C91459F Unknown NA C91462F Unknown NA C91467F U"kn0Wn NA C91473F UOlrnOWn NA C91479F U"lrn0W NA C9148OF unknom NA C91485F U"knOW0 NA C91487F lJnkrL0wn NA C91488F UnhlOW NA ale orrected PFOS LOQ (0.0 PFOSA = Pcrfluo-clancsvlfonamide P m s u = Pemuo-ctanesuironamidolcetale -new LOQ is 0.0198 ugiml A data is above the uppe EIFOSE = N m w RangeN-Ethyl Pertluomoamesulfonunido dhyl alcohol M556 = C8F17S02N((H)CH2C00) - PFOSAA - Dllulion FIdO, 50 50 50 50 50 10 50 50 50 50 50 50 50 50 50 - 50 50 20 20 20 20 20 20 20 20 20 20 IO 20 20 10 20 20 20 - 20 20 1 I I I I I I I - I I I I 1 I I I I I I I - I I ug/mL) 10 AC 021191( lil ofqumtl - PFOSAA - Canc. "ghL 132 154 67 I 475 I77 119 193 329 134 279 174 84 112 207 71 0 - 251 I73 510 570 94 I 922 498 559 529 553 555 596 602 562 688 776 624 670 648 - 515 646 0 00 0 00 ow OW 000 OW OW OW - 000 000 26 I OW 000 000 OW OW ow OW 000 OW - OW OW ude rtd co Do81600055 A022800018 A022800019 A022700038 Do81600058 W81600038 A022800021 A022800024 A022800025 A0227W040 DO81600059 Do81600060 A022800026 A022800027 W816W061 D081M)(X162 A022800028 0062000061 D062000062 w62oooO65 D062WMM6 D062000067 w62oooO68 DO62W69 D062oooO72 DM2000073 D06200W74 A053 1 0029 DO42000076 D062W79 A05310032 D062W0081 D062oooOBZ W62oooO83 DO62000086 w62oooO87 A062000022 4062oooO23 4062100026 4062100027 4062100028 4062100029 4062100030 4062100033 4062100034 4062100035 4062100036 %?62100037 4062100040 4062 loo04I ~mioo042 \0621oo043 to62 I Mo44 W-52100047 ~0621oo048 \062100049 ,\062100050 !\O62l 0005 1 lion facton -- Concentration -- olPFQSAA u#mL or % Ree 6 58 7 68 3:; 6 29.7 % 84 1.19 964 Ill5 . 6 59 14 0 8 71 4 19 5 58 10.3 3 :is -- 12.6 8 63 10.2 11.4 18 8 18.4 IO 0 II 2 106 II I II I 119 6.02 II 2 13 8 7 i6 I2 5 I3 4 I3 0 -- 10 3 I2 9 .LOQ (0.0248 ug/mL) LOQ (0.0248 ug/mL) LOQ (0.0248 ug/mL) LOQ (0 0248 ug/mL) .LOQ (0 0248 ug/mL) .LCQ (0 0248 uplmL) LOQ (0 0248 ug/mL) LOQ (0 0248 ug/mL) -- LOQ (o.0~48ugimL) LOQ (0.0248 UpimL) 0.0261 LOQ (0 0248 uplmL) LOQ (0.0248 u@L) LOQ (0 0248 ugimimL.) LOQ (0.0218 ug/mL) LOQ (0.0248 ug/mL) LOO (0 0218 ue/mL) LOQ (0 0218 ugimL) LOQ (0 02t8 ug/mL) LOQ (0.0248 U g h L ) -- LOQ [n 02.!8 ugimL) LOQ (0 0248 U g h L ) on and is provided as an enimale PFOSEA = Perfoorwelane sulfonyl elhylvnide Average PFOSAA uglmL 10 7 II 9 <LOQ RSD Sld. Dev. MSIMSD RPD 74 I 7.93 25 4 3 02 NA NA NA NA Dale Entered/By Dale Verified By Purity EnIercdNerified. 03106i00, 0311OlW.03120100, 03121100,4115lW. 5116i00. 06113100. 6/15/00,6/19/00, 06122100. 06123100, 04'26100,08115100, 09105100. 10;17/W LAUCSH 04130101 hoj 104130101 LAC OUl9lOl LAC 3M EnEETxvcSe-li89-r57.oI nmental Laboratory Sera Week 105 (3) TOX-00I-sera-212-1IAl.xls Page 51311204I 243 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance# 6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 Study 104 Week Dietvy Carcinogenicily Study with Nvmw Range (98 I%) N-Ethyl Pduomctanesulfonamido Ethanol i i Rals Product NumbeflTesI Substance): M&X T-6316 (EIFOSE4Xi) RaI Serum McthcdlRcvisian ETS-8-61 & ETS-8-5.1 Analylical Equipmoll System Number lnsmmcnl SoftwarcNersion Soup 020199, Davey 070799 Masslynx3.3 Br 3.4 Filename. See Below R-Squued Value: See Attachmcn~ Slope See Attichments Y-l"lereep1: Dates of ExUacliodAndysi. See Allaehments 02107100, 02108100,02109100 RWW,SAL Dates of AndyridAnalyst OZll8l00. U28100,03127100, 0513ll00, 06119100.06120100, 08103100, 08116100,08123100 UMHAAS Dale of Ddla ReductiodAnalysl: 02121100, 02129100, 3102100,06'02/00,06120100. WZllW, 08107100, 08118100, 08125100 IUMWlASMOJ Sample Data Box 00-009 WEEK 105 RAT SE lot UnknowniSWl3 Group Sample )I EtPOSE-OH Purity Dose Correction Pactor Group 4 C909il4M U"kn0Wn NA Mid-Hi& Dose C90908M Unknown NA Dilution FldW COK. Filename A080300017 A080300018 -conrmtr~tion -ef EIFOSE-OH <LOOQym(0L000197%7 uagc/emL) 0 0126 Immg/kg WIOM C90911M C90916M C90926M CW927M C90928M C90933M C90944M C90947M C90948M C90953M C90955M C90956M U"hlOwn U"kn0wn U"kn0Wn U"lol0Wl U"kn0Wl U"knOw0 unknown Unknown U"kn0W U"kn0Wn U"kn0wn UIlblOWl U"kn0wn NA NA 2nd Analysis OK NA NA 2nd Analysis OK NA NA NA NA NA NA NA 81 6 A080300019 00816 1 4 19 A080300020 <LOQ (0 00977 uglmL) I 22 I W81600053 0 0221 I I 0440 A0803W024 <LOP (0 00977 uglmL) I I6 4 A080300025 00164 I I 227 DO81600054 0 0227 I II 2 A08030W27 00112 I 11.6 A080300028 0 01I6 I 7 73 A08030003I <LOQ (0 00977 ug/mL) 1 6 26 A080300032 <LO) (0 00977 ug/mL) I 21.7 A080300033 0.0217 I 15.4 A080300034 00154 I I28 A080300035 <LOQ (0 00977 ugimL) C90965M Unknown NA C90968M U"kn0Wn NA A0803W038 A080300039 00197 0 0164 Groop 4 Mid-High Dose lmmg/kg C91315F C91324F C91327F C91332F C91335F C91336F C91338F C91340F U"kn0wn U"lmOW7l UlllmOWn unknown U"hlOwn unknown U"kn0Wn UllknOWl 2nd Analysis OK NA NA NA NA NA NA NA I1 W81MMo47 I 344 A080300041 I 29 0 A080300042 I 31.2 A080300045 1 56.2 A080300046 1 30.0 A080300047 I A080300048 I A080300049 0.0672 0.0344 0.0290 0 0312 0.0562 0 0300 0 0274 00101 Group 6 Mid-High Dose Recovery lmmg/kg Gramp 6 Mid-High Dose RCCOWry 100 mglkg C91341F C91342F C91356F C91357F C9136OF C91361F C91362F C91372F C91376F C91378F C91380F C91047M C91054M C91056M C91060M C91065M C91070M C91071M C91072M C91073M C91076M C91451F C91453F C91455F C91459F C91462F C91467F C991473F C91479F Unknown U"kn0wn U"kn0Wn Unknown U"kn0W U"knOw0 Unknown UllknOWn U"kn0W NA NA NA NA NA 2nd Analysis OK NA 2nd Analysis OK NA 2nd Analysis OK NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA 1 43 6 A0803WO52 0 0436 I A0803W053 0 0337 1 I 17.8 A0803wO54 I 33 5 A0803W055 1 57 3 A080300056 I 45.9 D081MMo48 I 170 A080300360 0.0178 0.0335 0 0573 0 0459' 0.0170 1 DO81600051 0 0467 1 A080300062 0 0408 - I 42 4 W81MMOSZ 0 0424 I 270 A080300066 0 0270 1 0 00 A062100022 ELOCI (0.00977 uglmL) I 000 A062100023 <LOCI(0.00977 ugimL) I OW A062100026 <LOCI(0 00977 ugimL) I OW A062100027 <LOCI(0 00977 uglmL) I 000 A062100028 cLOC (0.00977 u&L) I 000 A0621W029 <LOCI(0.00977 ugituL) I 000 A0621OOO30 c L N i (0 00977 ug/mL) I OW A062100033 E L K ' (0.00977 uglmL) - 9 1 000 A062100034 cLOC (0.00977 ugimL) I A062100035 (0.00977 u&L) I 0 0800 A062100036 cLW (0 00977 ugimL) I 000 A062100037 CLOQ (0 00977 uglmL) I 0 00 A062100040 <LGQ (0 00977 ugimL) I 0 00 AM2100041 <LO9 (0 00977 ug/mL) I OW A062100042 ELOQ( 0 00977 ugimL) I 000 A062100043 cLW (0 00977 ugimL) I ow A0621MX)44 ELOQ ( 0 w977 U g i r n L ) I OW A062100047 <LOQ (0.00977 ugimL) C91480F C91485F cLOQ (0 00977 uglmL) ELOQ(0.00977 uglmL) C91487F C91488F PFOS = Pcrfluomoctanesu ate PFOSA = Pemuorooclanerulfonaide PFOSAA = Pemuorooclanesulfonamidoaeelate NA orrectedP M S LOQ (0.0247 uglmL) la inelude std correctionIxtors new LOO is 0 0198 upimL. LAC 02119101 A - data is above the upper limit ofquanlilalion and is provided is an estimatc. -cLOQ (0 <Lop(0 00977 00977 u&L) uglmL) EIFOSE = Nmow Range N-Ethyl Pc~uaroaclanesvlfonatnidoethyl alcohol MS56 = C8FI7S02N((H)CHZC00) PFOSEA = Perfuorooctane sulfonyl cthylamidc Dale EnlerediBy. Date VerifiedI By' h r i l y EntcrediVerified miwm, O~IIOIOOM, I~OIOO,~IZIIO4Oi,isiw. 5 w w . wi31o0,6/151w, W I ~ I W0,6mm.~23100,06126100,msiw. WIO~IWI,OII~IM)LAUCSH 04130101 hoj 104130101 LAC 02119101 LAC Average EtFOSE-OH llghL 00182 0.0364 <LDQ <LOQ RSD Std. Dw. MSiMSD RPD 93.3 0 0170 40 0 00145 NA NA NA NA 3M EnEExTvcSe-li89-r57.oI nmental Laboratory Sera Week 105 (3) TOX-001-sera-212-1INXIS Page 244 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covsncell6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 Study Product Number(Test Substmce). MaviX: MethodiRcvision. Analytical Equipment System Number. lnsmmcnl SoRwueNcrrion: Filename. 104 Week Dietary Carcinogenicity Study with Narrow Range (98.IX) N-Ehyl PerfluorwclulesulfonamidaEhmol in Rals T-5316 (EffOSE-OH) Rat Serum ETS-8-4.1 & ETS-8-5.1 Soup 020199, Davey 070799 Masslynx 3 3 & 3.4 See Below R-Squared Value. slope: Y-lnlercept. Dates of ExUacliodAnalysl Dales of Analysidhalysl See Altachmenls See Attachmcna See Attachmenu 02/07/00, 02/08/00. 02/09/00 RWW, SAL 02/18/00, 2/28/00, 03/27/00, 05/31100,06/19/00. 06/20/00, 08103100,08/16/00, 18/23/00 MMWIAS Dale of Data Reductiodhalysl. 02/21/00, 02/29/00, 3/02/00,06102/00. 06/20/00. 0612IIW. 08/07/00,08/18/00. #)8/25/W MMWIASHOJ Sample Data Box O M WEEK 105 RAT SE GlOOP Sample I) lolNB113047-80 M556 Purity Sllrrogpte ~ 5 % -- M556 Filename -Concrnlrrtlon Doll Group 4 C90904M Corrcctlon Factor U"kn0wn Verified NA Dilmtlon Factor IO -- COW. aglmL 646 A022700090 - of M556 iiglmL or % Rcc 6 46 Mid-High Dose C90908M U"kn0Wn NA IO 766 A022700091 7 66 IO0 m a g C90910M Unknown NA 50 295 A022800019 14 8 C90911M U"h0Wn NA IO 610 A022700054 6 IO C9G916M Unknown NA IO 599 A022700055 599 C90926M U"lm0W NA IO MI AO227W095 6 41 C90927M unhown NA IO 824 A022700396 8 24 C90928M Unknown NA IO 694 A022700097 6 94 C90933M UnknOwO NA IO 899 A022700098 8 99 C90944M UDknOWI NA IO 410 A022700356 4 IO C90947M unlmown NA IO 416 A022700059 4 16 C90948M U"kn0Wil NA 10 538 A022700060 538' C90953M U"lmOw0 NA IO 401 A022700101 4 01 C90955M Unknown NA IO 871 A022700102 8 71 C90956M U"kn0Wn NA 10 345 A022700061 3 45 C90965M U"kn0W NA C90968M U"kn0W NA C91315F U"kn0WD NA -~- I O 42 I AO227W062 4 21 IO 627 A022700103 6 27 IO 635 A02270W77 6 35 C91324F Unkn0Wn NA IO 42 I A022700080 421 C91327P U"hOW7l NA IO 320 A022700081 3 20 C91332F U"hOw0 NA 20 288 A022800048 5 76 C91335F U"h0Wn NA IO 570 A0227WO83 5 70 C91336F Unhown NA IO 753 A022700384 7 53 C91338F U"lm0Wn NA IO 608 A0227WO87 6 08 C9134OF U"kn0Wn NA IO 388 A022700388 3 88 C91341F U"knOW7l NA IO 505 A022700089 5 05 C91342F Unknown NA IO 618 A0227W063 6 18 C91356F U"kn0Wn NA IO 210 A022700366 I IO C91357F U"kn0WlI NA IO 625 AO227W067 6 25 C9136OF U"knOW7l NA IO 460 A0227W068 160 C91361F U"kn0W NA IO 305 A022700069 3 05 C91362F U"kn0Wn NA IO 512 A022700070 5 I2 C91372F U"kn0Wn NA IO 606 A022700073 606 C91376F U"lm0W NA IO 620 A022700074 6 20 C91378F Unknown NA C91380F U"kn0Wn NA C91047M U"kn0W NA -. - IO 376 A022700075 176 IO 512 A022700076 5 12 I 0.0600 A061900022 LO7 (0 00494uglmL) C91054M NA 1 0.00 A0619WO23 LOQ (0 00494 ug/mL) C91056M NA I 0.00 A061900026 LOI) (0 00494 uglmL) C91060M NA I 0.270 A061900027 LO) (0 00494 ug/mL) C91065M NA I 0 00 A06 I W 2 8 LO) (0 00494 ugimL) C91070M NA I 0 00 A061900029 LO) (0 00494ugimL) C91071M NA I 0.00 A 0 6 1 W 3 0 L a > (0 00494 u@mL) C91072M NA I 4 41 A061900033 LO) (0 00494 uglmL) Group 6 C91073M C91076M C91451F .- - NA I 0.00 A061900034 Lo() (0 1W94 uglmL) NA 1 000 A061900035 LO> (0 00494 ug/mL) NA 1 55 3 A061900336 LOO (000494 uglmL) Mid-Higb Dose C91453F U"kn0Wn NA I97 A061900037 L a ) (0 tK1494 ugimL) Recovery C91455F Unknown NA 0.350 A061900040 LOO (0 w494 ugimL) 100 mgitg C91459F Unbwn NA 0.00 A0619wo41 L a ) (0 W 9 4 udmL) C91462P U"kn0Wn NA 1.87 A061900042 LOO (0 00494 ugimL) C91467F U"kn0Wn NA OW A061900043 LOO (0 00494 ugimL) C91473F U"h0Wn NA 0 350 A0619000d4 LOO (0O M 9 4 ur/mL) C91479F U"h0Wn NA 0.00 A061900047 LOO (0 00494ugimL) C91480F C91485F C91487F C91488F Uolmown UllhWn Unknown UIhOWn I NA NA 1 NA NA I 0.00 A061900048 LOO (0 00494 ug/mL) 2 75 A061900049 LOO (000494 ug/mL) 2' OW A 0 6 1 W 5 0 LOO (0 00494ugimL) 0.00 A061W51 (0 00494 "g/InL) FOS = Pemuomocianesu~ ate onected PFOS LOQ (0 0247 uglmL) to in l C std COrreCl U'IOTS FOSA = Perlluomoclmer inamide ncwLOQisO0198uglmL LACOU19I01 FOSAA = Perfluomoctul fonamidaacetate A -data is abave the UDM limit ofaumtitation and is vmvidcd as an estimate EtFOSE = Narmw Range N-Ehyl Pcrfluomoctanesulfonamidoethyl alcohol M556 = C8Fl7SOZN((H)CHZC00) PFOSEA = Pcrfuomoclmcsulfonyl elhylmide RSD Sld. Dev. MSMSD RPD 41 2 2 71 27 5 I 39 NA NA NA NA Date EnteredlBy. Date Verified/ By Purily EnleredNerified 03106100,03110100,03/ZO/W,03/21100.4/I5/W, 5/16100,06/13/00. 6115/00.6119/W. 06/22/00. 06123100, 36126/00.08/15/W, 09/05/0l). 10117100LAWCSH 04/30/01 hoj /04/30101 LAC 02119101 LAC 3M EnEETxvcSe-li89-r57oI nmental Laboratory Sera Week IO5 (3) TOX-001-sera-212-1IN XIS Page 245 3M Medical Department Study: T6316.1 AMDW 092597.1 Covance# 6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 Study Fmducl Number(Tes1 Subslance). MZlriX MethodRevirion Analytical quipmcnt System Number: Instrument SoflwarrNersion: Filename. R-Squared Value. Slope: Y-lntereepl' Dales of EmactioniAnalyn Dales 01AnrlyddAnalyst: Dale of Dala ReductiodAnalyst. Sample Data WEEK 105 RAT SI Gl0"P Sample # Group 4 Mid-High Dose IWmgkg Croup 6 Mid-High Dose RCC0"eV IW mgikg CWMM C90908M C90910M C90911M C90916M C90926M CW27M C90928M W33M CWMM C90947M CW948M C90953M C90955M C90956M C90965M C90968M C91315F C91324F C91327F C91332P C9133SF C91336F C91338F C9134OF C91341F C91342F C91356F C91357F C9I3M)F C91361F C91362F C91372F C91376F C91378F C91380F C91M7M C91054M C91056M C91060M C9106SM C91070M C91071M C91072M C91073M C91076M C91451F C91453F C91455F C91459F C91462F C91467F C91473F C91479F C9148OF C9148SF C91487F C91488F 104 Week Dietary Carcinogenicity Study wih Narrow Range (98.1%) N-Ehyl Pem,orooetvlcsulfonamido Ehilool in Rae T-6316 (ElFOSEeH) RaI Serum ETS-84 I & ETS-8-5 I Saup020199,Davey070799 Maslynx 3.3 & 3 4 See Below See AuachmenS See AUachmenS See Anachmenls 021071W. 02/08/00. 02109100 RWW. SAL 021181W, 2128100, 03127100, 051311W. 06119100. 06120iW.08103100, 08116/W, 08123.W W A S 02121/W,02129/00, SIOZIW, 06/02/W.06/2OlW.W211W.081071W. 08118lW. 08125.00 MMHAAS/HOl Box W-009 lo1 529 PFOSEA Purity Correction Unknown U"kn0Wn U"kn0W U"kn0wn Unknown Snrrogatc PFOSEA Verlfled Dllntlor Factor NA I NA I NA I NA I 2nd Analysis OK I NA I NA I 2nd Analysis OK 1 NA I NA I NA I NA I NA I NA I NA 1 NA I NA I 2nd Analysis OK 1 NA 1 NA I NA I NA I NA I NA I NA I NA 1 NA I NA I NA 1 NA 2nd @alpis OK NA 2nd Analysis OK NA 2nd Analysis OK NA NA NA NA I NA I NA I NA NA NA NA NA NA NA NA NA NA I NA I NA 1 NA I NA I NA I NA 1 NA orrecled PFOS LOO 1 riGzI Eaz new LOQ is 0 0198 u>mL LAC 02119101 A. data is above the upper limil of quvllitalian 0.W OW OW 0.04 OW 0.W 0.W 3.75 OW OW 4 02 6 88 0.00 I85 0.320 OW OW OW OW I72 OW 2 40 I61 OW 0610 6 20 104 OW 0 00 OW OW OW OW OMX) OW OW OW OW OW OW OW OW OW OW OW OW 000 OW 0.W OW rtd correction aod is provided I A080300018 A080300027 A08030W28 A080300031 A080300032 A080300033 A08030W34 A080300035 A080300038 A080300039 D0816W047 A0803W041 A0803W042 A0803W045 A0803W046 A0803W047 A0803W048 A0803W049 A080300052 A080300053 A080300054 A080300055 A080300056 W816W048 A0803WOMI W8 I6oW5 1 A0803MH)62 W81600052 A080300066 A061900022 A061900023 A061900026 A061900027 A061900028 A061900029 A061900030 A061900033 A061900034 A061900035 A061900036 A061900037 A061900040 A06190004I A0619W042 A061900043 AM19000M A0619ow47 A061900048 A0619W049 A061900050 A061900051 lOrS an estimate -- co,,cm1ration -- or PFOSEA "gl"lL or % Rce cLOQ (0.00492 ug/mL) cLOQ (0.00492 ugimL) <LOQ (0 00492 ug/mL) <LOQ (0.00492 ug/mL) <LOQ (0.00492 ugimL) <LOQ ( 3 00492 ugimL) CLOQ (1 00492 ug/mL) <LOQ (I)w492 ug/mLl <LOQ (I) 00492 ug/mLl <LOQ (000492 ug/mL) <LOQ (0.00492 u&L) <LOQ (11,00492 ug/mL) <LOQ (1100492 ug/mL) cLQQ ( 0 OM92 ug/mL) <LOP (ll.W492 UgimL) <LOO (11,00492uglmL) (I w 6 8 8 cLGQ ((I 00492 ugimL) <LOQ (000492 ugimL) <LOQ (11 00492 ugimL1 <LCQ ([I 00492 uglmL) cLOQ (11 00492 ugimL) <LOQ (CI 00492 ug/mL) <LOQ (C 00492 ugimL) <LOQ (C 00492 ug/mL) cLOQ (0 00492 ug/mLl <Log (0 W Y Z ugimL) <LOQ (0 00492 UgimL) <LOQ (0 00492 ugimL) <LOQ (0 00492 ugimL1 0 w620 -- <LOQ (0 OM92 ug/mL) <LOQ (0 OM92 ugimL) cLOQ (0 00492 ug/mL) <LOO (0 00492 ugimL) <LOQ (0 00492 u&L) cLOQ (0 00492 ugimL) <LOQ (0 00492 ug/mL) <LOQ (0 00492 ug/mL) <LOQ (0 00492 ug/mL) ELOQ (0 00492 ugimL) -- <LOQ (0 00492 ug/mLl <LOQ (0 00492 ug/mL) cLOq (0 OM92 ugimL) cLOQ (0 00492 ug/mL) <LOQ (0 00492 ug/mL) <LOQ (0 w492 ug/mL) <LOQ (0 OM92 ug/mLl <LOQ (0 00492 ugimL1 cLOQ (000192 up/rnL) cLOQ (0 00492 u&L) CLOQ (0 1x492n@mL) <LOQ (000492 UgimL) -- cLOQ (0 w492 ug/mL) cLOQ (0 30492 ugimL1 Average PFOSEA UgllnL <LOQ <LOQ RSD Std. De". M W S D RF'D NA NA NA NA NA NA NA NA Date EnIerediBy. Dale Vcnfiedl By: Purity EnImedNcrified. 03106lW. 03110iW. 031201W, 031211W.41151W. Sl16/W. W131W. 611SlW. 6/191W, 06/22100.06123/W. W26,00,081151W. 091051W. IO! 17/00 LAWCSH MI30101 hoj 104130101 LAC 02119101 LAC 3M EnEExTvcSe-li89r-57.o1 nmental Laboratory Sera Week 105 (3) TOX-001-sera-212-1IAI XIS Page 513ll2W I 246 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance# 6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 Study Roducl Numbcr(Tes1 Substance)' MauiX 104 Week Dietary Cvcinogmicity Study with N m w Range (98 I%) N-Ethyl Pemuomoclviesulfonamido Ethanol in RIIS T-6316 (EIFOSEOH) Rat Serum MethodiRcvision. ETS-84.1 & ETS-8-5.1 Analytical Q u i p m e 1 System Number Soup020199,Davey070799 lnsuument SaRwardVcnion: Filename. R-Squared Valuc. Masslynx 3.3 Br 3.4 See AttaehmmU See Auachmmet Slope. See Atwchmcnts Y-Intercept. Dates o f ExhactiodAnalysI- See Attachmenet 021071W, 02108lW. 02/09/00 RWW, S A L Dates of AndysisiAnalyst 02118100, 2/28100,03127100, 05131/00,06119100, 06120100,081031W.081161W,08/23/00 MMHllAS Dale of Data ReducliodAnalvst. 02121100,02/29100. 3/02/00. 06102100, 0612OlW. 06121100. 08107100, 08II8100. 08125100 MMHilASlHOJ Sample Data WEEK 105 RAT SE Group 4 Sample X C90904M Conceatration of PFOS mglmL or % Rec 66 9 - Avcragr PFOS IIglmL RSD Std. De". MSIMSD RPD Concrntrrtian of PFOSA tlglmL or % Ree 0 568 AWP@ PFOSA uglmL RSD SId De". MSIM? Mid-High Dose C90908M 71.0 0 606 100 mg/kg C90910M 313 I84 C90911M 146 0 607 C90916M 88.4 0.612 C90926M 61.61 0 330 C90927M 22.0 I09 C90928M 127 0 979 C90933M 45 8 C90944M 136 0 593 0.560 C90947M 144 0.685 C90948M 51.7 0 277 C90953M 13.0 0 572 C90955M MI C90956M 132 0614 0 390 Gloop 4 C90965M C90968M C91315F - 137 53.6 100 84.6 45 4 122 0 949 0 638 0.898 0 701 519 0 %1 Mid-Htgh Dose C91324F 133 1.91 C91327F 9 81 I .02 C91332F 251 A 1.36 C91335F 99.5 2 05 C91336F 202 A I07 C91338F I 40 1.71 C91340F 252 A 0 721 C91341F 76.0 I38 C91342F 122 0 826 C91356F 6.18 0 345 C91357F 200 A 2 01 CY l360F 141 I85 C91361F 131 I16 C91362F 25.3 0 720 C91372F 58.8 I61 Graop 6 C91376F C91378F C91380F C91047M 133 IO9 104 0 185 - 122 57 9 70 5 I 93 I17 cLOQ (0 OM93 UgimL) Mnd-High Dose C91054M 0.0222 .LOQ (0 00493 "g/"lL) Rccavery I00 "@kg C91056M C91060M 1.18 0 142 CLOQ(0 00493 up/mL) :LOQ ( 0 OM93 u&L) C91065M C91070M C91071M :LOO (0 0198 ug/mL) 00514 :LOQ (0 0198 ug/mL) cLOQ (0 00493 ug/mL) CLOQ(0 00493 ug/mL) CLOQ(0 00493 uglmL) Group 6 Mid-High Dose C91072M C91073M C91076M C91451F C91453F 0 0454 ZLOQ (0.0198 UglmL) 0 592 5.02 I68 - 0 227 166 0.377 rLOQ (0,00493 ug/mL) cLOQ (0.00493 ug/mL) cLOQ (0 00493 u&L) :LOQ (0.00493 ug/mL) :LOQ (0.00493 ug/mL) --NA NA Recovery 100 mglkg C91455F C91459F 5 92 :LOQ (0.0198 ug/mL) :LOQ (0.00493 ug/mL) :LOQ (0 OM93 ug/mL) C91462F C91467F C91473F C91479F 4.61 0.0207 3 84 :Lop (0.0198 ug/mL) :LOQ (0 OM93 ug/mL) ZLOQ(0 W493 ug/mL) rLGQ (0 00493 Ue'rnL) :LOQ ( 0 00493 ugimL) C9148OP 0.0553 :LoQ (0.M93 uglmL) CY1485F 3 55 rLOQ (0.00493 ug/mL) C91487F ZLOQ (0.0198 u&L) 96 3 :LOO (0.00493 up/mLl N, ! C91488F 4.16 2.41 2 32 PFOS = Pcrfluorooclanesul ate PFOSA = Perfluorooctanesulfonvnide PFOSAA = Pc~uorwetanesulfonimidoacctlte cc :led PFOS mL new LOO is 0 019 A - damis above U (0 0247 ug/mL) I EtFOSE = Narmw Rangc N-Ethyl Pemuomctancsulfonamido ethyl alcohol M556 = C8F17S02N((H)CH2C00) PFOSEA = Perfuorooctane sulfonyl cthylamide Dale Entcred/By Dale Verified/ By Purity EnIerediVerified 03106100, 03110100,03120100,031211W,4llSlW. 5116AW. O6113lW. 61lSIW. 6119100,06/221W. 06123100. W26/W, 08115l(IO. 09lOSlW. 10117lW LACICSH 04130101 hoj I04130101 LAC 02119101 LAC ConerntrPllon 01 PFOSAA mglmL or %e Rec 6 58 7 68 33 6 23 7 8.8 I19 964 I6 5 6 69 14.0 8.7 4 19 5 58 10 3 3 55 12.6 8.63 IO 2 I1 4 I8 8 18.4 10.0 I1 2 10.6 11.1 I1 I 11.9 6 02 11 2 I3 8 7 76 12 5 I3 4 I3 0 1,.,0_.93 , LOQ (0 0248 ug/mL) LOQ (0 0248 ug/mL) LOO (0 0248 ug/mL) LOQ (0 0248 ugimL) LOQ (0 0248 uglmL) LOQ (0 0248 ugimL) LOQ (0 0248 ug/mL) LOO (0 0248 ug/mL) LOQ (0 0248 ug/mL) LOQ (0 0248 ug/mL) 0 03 LOQ (0 0248 ug/mL) LOQ (0 0248 ug/mL) LOQ (0 0248 uglmL) LOQ (0 0248 uglmL) LOQ (00248 UgimL) LOQ (00248 us/mL) LOQ (0 0248 ug/mL) LOQ (00248 ug/mL) LOQ (0 0248 ug/mL) LOQ (0 0248 ug/mL) LOQ (0 0248 ug/mL) RSO Std. De". MSIMSD RPD - IO 7 MI 7 91 25.4 3.02 NA NA NA NA 3M EnEETxvcSe-il89-r57.o1 nmental Laboratory Sera.Week 105 (3) TOX-WI-scra-212-IIAI XIS Page SI3112wI 247 3M Medical Department Study: T6316.1 AMDW 092597.1 Covancel) 6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 Study Pmducl NumbcNTest Substance). 104 Week Dietary Carcinogenicity Study with Nmow Range (98 I%N)-Ethyl Pnfiuorwclanesulfonmido Ethanol in Rats T-6316 (EIFOSEOH) Matrix: MethodiRevision Analytical Equipment System Number InSUumcnt SoRwarcNcrrion. Filename R-Squared Value. slope: Y-lntercepl Dates of ExUactiodAnalyn. Daln of AnalysidAnndysl Rat Semm ETS-8-4 I & ETS-8-5.1 Soup 020199. D a v q 070799 Masslynx 3.3 & 3.4 See Below See Attachments See Allachmentr; See Attachments 02/07100,02/08100, OZlO91W RWW. SAL 02118100, 2/28/00, 03121100, 05/31100,06119100,06/20100. 081011100, 08116100, 08/23/00 MMWlAS Date of Data Reduetionihalyst 02IZllO. 02129lW. 3/02/00, 06/02/00, 06120100, 06121100, 08107100,08118100. 08125100 MMWlASfllDJ Sample Data BOX 00-009 WEEK105 RATSE ClOUP DOSC Sample # Corncentration of EtFOSE-OH uglmL or % Ree Average EtFOSE-OR ughL RSD Std.Dw. MYMSD RPD CO"Ce"t*atiO" 01M556 aglmL or Rec Group 4 C90904M :LOQ (0 00971 ugirnL1 6.46 Mid-High Dose C90908M 00126 7.66 . WQmgkg C90910M 0 0816 14 8 C90911M cLOQ (0 00977 ug/mL) 6.10 C90916M 0.0221 5.99 C90926M :LOQ (0.00977 udmL) 641 C90927M 00164 8.24 C90928M 0.0227 6 94 CSil933M 0.0112 8.99 C90944M 0 01I6 4 IO C90947M cLOQ (0 00971 ugirnL) 4.16 C90948M :LOO (0 00977 ug/mL) 5 38 C90953M 00217 4 01 C90955M 0.0154 8.71 C90956M :LOQ (0.00917 ugimL) 3.45 crollp 4 Mid-High Dose C90965M C90968M C91315F C91324F C91327F 0.0197 00164 0 0672 0 0344 0.0290 93 3 4.21 0.0182 00170 6.21 6 35 421 3.20 41 I 8 2 71 C91332F 0 0312 5.76 C91335F 0.0562 5.70 C91336F 0 0300 1.53 C91338F 0.0274 6 08 C91340F 0 0101 3.88 C91341F 0 0436 5 05 C91342F 0.0337 6 18 C9 I356F 0 0178 2.10 C91357F 0.0335 6 25 C913M)F 0.0573 4M) C91361F 0 0459 3.05 C91362F 0.0170 5.12 C91372F 0 0467 606 C91376F 0 0408 6.20 C91378F 0.0424 40.0 3.76 27.5 C91380F 0 0270 0 0364 0.0145 5 I2 1 xq Group 6 Mid-High Dose RCCOWIY 100 m g i g Croup 6 Mid-High Dose Rccovery loomfig C91047M C91054M C91056M C91060M C91065M C9 I070M C91071M C91072M C91073M C91076M C9 I45 I F C91453F C91455F C91459F C91462F C91467P C91473F C91479F C91480F C91485F C91487F C91488F :LOQ (0.00977 ugimL) :LOQ (0 00977 ugimL1 :LOQ (0.00977 ugimL) :LOO (0.00977 ugimL) :LOQ (0.00977 ug/mL) :LOQ (0.00977 ug/mL) :LOQ (0 00977 ug/mL) .LOO(0.00977 udmL) LOQ t0 00977 up mLj , IOQi000977ugmL, LOO 0 00977 "I rnl L& (0 00977 ug/mL) LOQ (0 00977 ug/mL) LOQ (0 00977 ug/mL) LOQ (0 00977 ugimL1 LOQ (0 00977 ug/mL) LOQ (0 00977 u&L) LOQ (0 00977 Ug/mLl LOQ(0 OW77 ug/mL) LOQ (0 00917 ug/mL) LOQ (0 00977 ug/mL) LOO (0 00977 ug/mL) <LOQ LOQ ( 0 00494 ugimL) LOQ (0 00494 ug/mL) LOQ (0 00494 ugimL) LOQ (0.00494 ug/mL) LOQ (0.00494 ugimL) M Q (0 00494 ug/mL) LOQ (0 00494 ugimL) LOQ (0 00494ug/mL) NA LOQ (0 00494 ugimL) NA LOQ (0.00494 ugimL) -OQ(0.00494 ug/mL) BQ(0.00494 ue/mL) LOQ (0 00494 ug/mL) LOQ (0.00494 ug/mL) -OQ(0 00494 ug/mL) LOQ (0 00494 ug/mL) LOQ (0 w494 "g/rnL) -OQ(0.00494 ug/mL) &Q (0 00494 uglmL) LOQ (0 00494 UgIrnL) NA -OQ(0.00494 ugimL) NA B Q (0 00494 ugimL) -- NA NA WC ,naniide fonaniidoacelale EIFOSE = Narrow Rangc N-Ethyl Pcmuorwclanesulfonamido ethyl alcohol ,mated PFOS LO -new LOQ is 0 0198, A data is above the 0247 ugimL) 10 inelud L LAC 02119101 cr limit ofquantilalian and is provided as an estimate. M556 = C8FllS02N((H)CH2C001 PFOSEA = Pcrluomwtane sulfonyl ethylmide Conerntrltio" of PFOSEA cLOQ (0 00492 ugimL) <LCQ (0 00492 ugimL) <LOQ (0 00492 ugimL) <LOQ (0 00492 ngimL) < L o 9 (0 00492 ugmL) <LOQ ( 0 00492 us/mL) CLOQ (0 00492 ugimL) (LOQ (0 00492 u&L) <LOQ (0 00492 ug/rnL) cLOQ (0 00492 us/mL) < M Q (0 00492 ugimL) <LOQ (0 00492 UgimL) cLOQ (0 00492 UgimL) <LOQ (0 00492 ug/mL) <LOO (0 00492 udmL) < L (0 ~00492 u&L) <LOQ (0 00492 UdmL) <LOP (0 OM92 u&L) 0 00688 cLOQ (0 00492 uglmL) <LOQ (0 00492 ugimL) <LOQ (0 00492 ugimL) <LOQ (0 00492 u&L) <LOQ (0 00492 ug/mL) (LOQ (0 00492 UgimL) <LOQ (0 00492 ug/mL) <LOQ (0 00492 ugimL) <LOQ (0 OM92 ug/mL) CLOQ (0 00492 ug/mL) <LOQ (0 00492 uglmL) ELOQ (0 00492 ugimL) <LOQ (0 00492 ugirnL) 0 W620 <LOQ,O00492upmL, cLOQ 0 00492 ug mL, <LOO i0.00492 &Lj <COQ (0 00492 u&L) cLOQ (0.00492 UgimL) CLOQ (0.00492 UgImL) ELOQ (0 00492 ug/mL) cLOQ (0.00492 UgimL) ELOQ (0.00492 ugimL) cLOQ (0.00492 UgimL) cLOQ (0 00492 uglmL) :LOQ (0.00492 UglmL) CLOQ (0.00492 ugimL) <LOQ (0.00492 UgimL) CLOQ (0 00492 ug/mL) FLOQ (0.00492 u&L) CLOQ (0.00492 ugimL) CLCQ (003492 up/rnL.) :LOQ (0.00492 ugirnL1 <LOQ (0.00492 ug/mL) :LOQ (0 00492 ug/rnL) :LOP (0 00492 up/mL) :LOQ (0 00492 ugirnL) Average PFOSEA odmL <LOP RSD Sld. De". MYMSD RPD NA NA NA NA NA NA NA NA Date Elcredkly. Ddle VWifiedI By Purity EntcrcdNerified 03106100, O3/10100,03120lW,03/21100.4115/00. 5llWW. 06113lW. Wl5100. 6/19/00. 06122100,06123100. 06126100,08115100,09,05100. 10117/00LAWCSH 04130101 hoj 104/30/01 LAC 02119101 LAC 3M EnEETxcvSe-l8i9-r57oI nmental Laboratory Sera Week 105 (3) TOX-MII-srra-212-IIAI.rls Page 248 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance 6329-212 EEK4 RATLIVERREWORK Lot 193 C90753M 09999 NA C10775M I0586 NA C91135F 1012. NA C91111F IO380 NA C91165F I0175 NA C91169F I0177 NA C9ll7lF 1- NA tiW"p2 CW789M 1.0591 NA Low hII CW814M I0571 NA 30m p ~ ~ CW817M I om NA CW831M I UIl8 NA CWSJIM I0749 NA C91198F I0531 NA C9lWJF 10651 NA C91222F I0183 NA C912JOF ,0433 NA 0 9275 0 9271 09275 0 9275 0 9275 0 9275 0 9175 0 9275 0 9275 u 9275 0 9175 0 9175 0 9175 0 9171 0 9171 0 9275 Unknown Unhown Uokmwn Unhown Unhown Unhown Unknown Unknown Unknown Unknown Unknown Unknown uoknown unknma Unknown Unknown 721 I 60398015 %3 I 60398046 221 I 60198050 284 I 603PSOJI 631 I 60398051 151 1 60398053 61 I 60398054 115 LM 60198111 120 IW 60538113 10487 10 5 112 IW 60598114 11723 I, 7 _ _ 169 lW 60598115 ISUll I5 0 ,lo IW 60598116 11069 125 100 60598119 ,1027 110 IW 60598120 9588 116 100 60598121 11282 106 IW 60598122 9412 941 13116 13 k 61510 6 15 86362 86 4 110931 111 121385 I 111 I91929 91 9 118167 138 151195 I51 118159 I I18 167159 102805 163867 M 211647 169257 169 ,19299 119 mom I 221 2,3655 92711 I91685 391682 MI557 691925 559479 559 557635 558 179688 380 17 I 113 I91 I I 1 I 21(I 110 25 1 I I 159 268 53 1 I I 499 Analytical Report: FACT-TOX-001 LRN-U2103 60598099 60598100 6059810, 60598101 60598105 60598106 60598107 60598108 6059BIW 0 670 0 636 0635 a651 0761 I 839 (10572 MOW298059 MUW298060 Mwo298061 M07W9813 I M0709981k M07WP815 M07W9816 M07w9817 MOIW9810 MOIW9821 I MU7099811 7099827 7099828 7099819 705983U 7099831 7099831 '099835 7099836 7099837 7099838 4761 5564 7036 9841 I 6639 8712 13084 9543 7302 1162 6181 11518 13154 11165 15517 18156 16151 11645 11836 6 60 5 93 741 4 16 5 57 7 04 9 81 6 61 8 72 13.1 9 5, 7 30 131 115 155 162 146 111 6 81 176 6 I4 108 1 1 114 9s i 133 56 5 8 94 5 05 IS I I5 3 231 3M Environmental ETSI-7 0 Ex-197 Laboratory Page 249 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance 6329-212 Sample Data C0.bol GTavD I REWORK s.mph L H20 Blk-il H20 BU-2 hbbil Lluwl3lk-l U b i , Livrr Blk.2 C90719M-MS" C90719M-MSD CW719M" CW725M C90732M C90753M C90775M C91115F CPII4IF C91165F C91169F C9ll7lF CW789M CPo811M C908I7M CW833M CW83kM C91198F FUlrm QFOSAA Cnk. C.n. "8k W39SUl4 385 61198056 ow 60398015 ow 61298017 OW-.- 60398041 131 60398113 I34 60398121 51 5 60398013 I05 I 60398011 143 I 6039800 I51 I 60398M6 119 I 60398050 113 I 6039805I 779 I 60398051 161 I 60398053 79.8 I 60398054 I15 IO 60598140 1087 IO 60598141 I9M IO 60598142 18k6 50 60598129 9731 10 60598144 2528 10 60598147 7681 10 6059814% 1751 10 64598149 3893 64598150 M59815l 1123 4320 90298049 97855 Po298050 54290 ::: 90298051 549uI IO 9 902980Ji 19263 90298051 59731 596 90298057 46021 46 0 W298058 93774 93 8 Po298059 25750 902980W 90198061 41154 mu6 M01099819 54589 MO7099811 M194 M07099815 ,1108 M07WPB16 88123 M07WPII7 56911 M07WY810 49148 MO70998ll 5368k M07WP829 Analytical Report: FACT-TOX-001 LRN-U2103 - EtFOSE EtFOSE C.W. D(lut*O 9b 32 5 F.UW I OW I OW I - .- OM I 19, I 133 I ow I OM I OM) I 30 7 I 0.w I ow I ow I ow I - ow I ow I 21 5 I ow I ow I - ow I ow I ow I ow I 69 2 I - om I OW I 128 I 45 7 I 6ko I 117 I _ _ 79 6 I 513 I Po298072 5, 3 49 I I W198073 49 I 448 I - I , 'I I 33 9 I 656 I 90298075 41 9 90298076 33 9 167 I 328 I 90298081 324 449 I 90298082 422 432 I Po198081 425 610 I 161 I 151 I - 124 I 117 I 375 I 496 I 1127 I - I M 5 I 551 I 926 I 1459 I 751 I - 364 I 90298105 361 371 I 90298106 166 00513 00491 00419 0 0339 0 324 0 122 0 125 0361 0 366 00442 0464 15 4 OW82 33 I 0 I54 "762 58 2 0 143 3M Environmental Laboratory Page 250 3M Medical Department Study: T6316.1 AMDT# 091597.1 Covrnce 6329-211 Analytical Report: FACT-TOX-001 LRN-U2103 I Grows - PFOS C-k.Cmc. 1u'I ll5 C90719WMSD C90719M" OM ~ I PI ow - 233 223 - 728 C90725M 647 C90712M 980 CW753M 670 C'H1775M CPlllSF 841 ~ 203 CPllllF 25, C9116SF 564 C9116PF C91171F C9078PM .02 - 418 12658 C90811M 10487 C90817M 11723 CPO833M lSOll C90834M C91198F 12069 ~ Ila27 C91205F 9588 C91222F 13282 - C9l2lOP 9.11 C91117F 13.26 131 C90846M 6,520 64 5 C90851M 86362 86. C90866M 110931 Ill CW890M 109767 110 C90891M C9125IF ~ 108824 99011 C91272F ,21315 C91285P 91929 91 9 - CP1117F 1111167 138 C91191F 151495 151 C'Hn35M 128159 128 CW939M 167259 167 CW942M 202805 203 C90%1M 263167 264 CWP66M 2,164, 211 ~ C91316F 169217 169 C91317F 4,9299 C91333F 22- C913LSF C91350F C90981M 2.3655 - 92721 191685 CILOO5M 391682 C91024M 661557 662 C91033M 691925 692 C91037M 559179 559 ~ C9L423F 557635 558 C91127F 379688 380 C91A3"F ,88915 489 C91443F C91448F - 816358 740764 rnl& mmidoaceYh 113 I91 21 2 721 7%l 5083 7159 7276 (I 712 7% 5 08 7 16 7 28 ,761 176 II 0 5566 5 57 110 25 3 7036 7 04 9841 9 84 6639 6M 8722 8 72 26 8 13084 13 1 I99 53 2 9543 9 54 I I I I I 73"Z 10851 I I 7 30 109 1362 I36 6181 6 IS 11518 115 41 7 13154 132 499 208 11465 115 15517 I55 18156 182 16152 16: - tI 682 I b00571 159 108 -97855 50940 I 39263 I 59731 46022 93771 33108 88123 56911 49148 0 161 0 0798 0 115 2 09 I94 I85 9 13 2 53 761 ow 19 2 ow 0 0995 0 "390 ow 22 1 ow ow 94 4 ow 3 63 3 42 ow I I I 000 :I 1 1 ; 393 59 7 396 22 6 79 6 .6 0 511 91 8 49 I 25 8 :t: 618 ::: 44 8 506 16 8 13 1 324 88 I 41 2 36 9 53 I 219 425 49 3 620 53 7 170 I I I 536 NA 738 283 NA 32 I 52 5 149 135 79 2 I I I 372 79 6 85 7 20 4 83 9 914 31 7 1128 29 6 739 I49 63 5 161 106 80 I 509 166 OOIlP 00339 0683 NA 0 321 0421 0 425 0620 n270 *A NA 0 235 0 372 of0 516 0914 0 719 0361 0 366 0 0442 I54 0 w682 33 I 0 464 0 I54 I I I I j 56 7 0 375 0213 I I 47 5 0 70, 10335 0 762 "58 2 443 3M Environmental ETSI-7 0 F X C d 97 Laboratory Page 251 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance# 6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 Study Product Numkrflest Substance) Ma& MelhodiRWlSlO" Analpica1Eqwpllenl System Number Lnsvwnenl SoRwareWenion Date ofExVacttOniAnalyS1 Date ofAnalyslslAnalylt Date oflhta ReduEt~onihlyst Sample Data 104 Week Dietary Carcmogenicq Shldy w P &'arrow Range (98 1%) 7-631 6 W O S E O H ) Rat LlVer FACT-M-I 0 & FACT-MJ 0 rewotked uine ETS-8-7 0 Cluck 080697 and Madeline 041098 MassL,m 2 3 and 3 I 06/01/98 RWWIOK 06112/98.06114/98.06/18/98.09102198 HOliKJWLAC 03116100, l1129100, 12/Ol/W, l2/101W IASMMHJKJH 06/14/98 Lot 193 Ohil8/98.12/29/98 K-Ehvl P e r f l t m m t m e s u i i o n d o E h o l rn Rats Filmame R-Squared Value Slope Y-Intercept b r Llst 10 hghl See Atllfhmenl~ Set Attachments Sea Attachments Filenames Blb G'p 1 Crp 5 M$MSD 06/12/98 LotL-2353 09/02/98 Lot L-2353 06112/98 Lo1617 06118198 Lot 617 PFOS PFOSA M06189834-35B6041 A90298W2-3 & 116-117 06189852.57, I22998069-7+?3298019-31 61498013-24 I 22998041-52 61298127-128 61298127-128 06/12/98 Lot936 09/02/98 Lot 936 PFOSAA M06189834-35 &a41 06189845.57 122998055-66 61298127-128 EtFOSE Dilutions A90298002-3 & I l l , 111, I l l . Ill IxI298019-31 111, Ill, 14, 111 need I l O & l IIIWW, l/lW, 1/1ooo, 1/10 90298110-111 n e e d l 5 & l , l i l , I / l , 1 / 1 HZO Bk-1 HZ0 BIk.2 Rabbit Liver Blk-l Rabbit Liwr Blk-2 C90717M-MS C90717M-MSD C90717M C90718M c90737M c9075m C90769M C91126F C91140F C91145F C91162F C91163F C90971M C90972M C90988M C90996M C91017M C91393F C91394F C91404F C91415F C91438F Inilia1 WI. I loo00 1WW I0080 I 0080 I0248 I0248 I0455 10268 1 0434 1 W89 1.W83 1 0205 I0265 10128 10160 10162 10129 I 0204 10108 10128 10116 1 0247 I0126 10149 I0244 10230 Total Mass of Liver I NA NA 40 13 40 13 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA 4/13/00, 5124/00CSH I2/lYWMMH 12/28/00 hoj (list a122998a) 12/26/00 LAC PFOS Sld Correction Fsrlor 0 9275 0 9275 0 9275 0 9275 NA NA 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9215 0 9275 0 9275 0 9275 0 9275 0 927s 0 9275 Lot 193 PFOSPurih I corrrrtian Factor UnknOUn unknown unknoun NA NA UnknOUn UnknOWn UnknOWn UhWl UnknOHm unknown unknown UllkllOUn unknown ClnknOwn UnknOUm UnknOWn UnknOWn UnknO\*n IlnknOwn unknown unknown Imnknoun LlnknOW 1inknnm PFOS w s C0"C. 47 4 OW 33 I 1153 959 313 222 307 218 I24 270 383 558 1198 M5 137 122 136 97.3 906 96 5 103 I12 116 IS7 I PFOS PFOS FilmnllX Dilullan CdC. cone. Fmbr WE OW 61298056 I 43 9 122998002 I OW 61298057 I 30 4 122998W3 1 -1584 61298127 I -1773 61298128 10 2776 122998073 10 2004 122998072 10 2731 122998071 10 2002 122998070 IO I144 122998069 I 245 6189852 I 346 6189853 1 511 6189854 1 1094 6189855 1 552 6189856 IWW 1250280 61498013 IWW 1111564 61498014 two0 1249941 61498015 loo00 890177 61498016 two0 830679 61498017 IWW 873644 61498020 IWW 945819 61498021 IWW 1027570 61498022 IWW 1054710 61498023 IWKC 1423345 61498024 Icalibrationm m ,estimated value + CCV's failed. &la enteredas estimated "confirmed low on 90298 ++ Qualitauve data only conrrntmuan or PFOS wvg or *ARec. <toQ(0 0110 "pip) <LoQ (0 112 U ! 3 w <toQ(0 0110 "pig) <roQ(oll2"pig) -1331% -14Wh 2 78 200 2 73 200 114 0 245 0 346 0511 I09 0 552 1250 1112 1250 891 831 874 946 1028 IO55 1423 M-0 PROS uvs <LW LoQ -1410% 2 13 0 549 1067 1065 RSD Sld. Dw. MYMSD RTD NA NA 11% 31 3 0 668 59 8 0 328 18 5 197 20 0 213 Ers-8-7 0 Exel 97 3M Environmental Laboratory Lw Week 8 Rework T0X-W I -liver212-100 XIS 513112001 Page 252 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance# 6329-212 study Sample Data 104 Week Dietary CarcinogenicityStudy w t h Narmw Range (98 1%)Y-Elhyl Perlluomtanesulfonamldo Ethanol m Rats T-6316 (ElFOSEOH) RaI Lwer FACT-M-I 0 & FACT-M-2 0 reworked usmg ETS-8-7 0 Filnume See LlS110 Rlghl Chick 080697and Madclme 041098 R-Squarrd Value See Atrachmenct MasrLym2 3 and 3 1 slaw See Atrachmenct 06101198 RWWIOK Y-Intercept See Attachments 06112198,06114198,06118198,09102198 HOJiKlHiIAC 03116100, 11129100, I2lOllW, l Z I l O i 0 0 I A W M W H WEEK 8 RAT LIVER REWORK SPrnPl. x Method B L M a m Blk H2O Blk-1 H2O Bk-2 Rabbit Lner Blk-1 l o t L-2353 PFOSA h"ly corrrrtion FWor C90717M-MSD 0 0 mgilrg c90737M C90756M Gmup 5 wgh Dose C91140F C91145F C91162F C91163F C90971M C90972M C90988M C91017M C91393F UhUm C91394F C91404F Unknown C91415F unknown C91438F PFOS = Perfluomamesulfonate PFOSA = Perfluomoctanesulfnnamlde PFOSAA - P e r f l u o ~ ~ t a n e s u l f o - d ~ ~ l ~ t ~ ElFOSE = N m w Range N-Ethyl Perfluomaranesulfonamldoculyl alcohol - C0"C n#i 00 - 6 6 3 00 41 8 - 83 4 65 4 - 33 8 196 35 2 31 I - 10 I 10 4 10 4 12 7 - 35 0 I5 2 I% 191 191 211 - 185 I54 142 I74 - 142 I77 - PFOSA - DiluUOn FnCtOr I I ~ I - - I I I I I I I - I I I I - 1 1 100 1W I00 IW - 100 IW IW 100 - I 0 0 100 41 4 19353 18747 18897 20877 14071 17148 13844 17272 Conrcntmtlon o f PFOSA uyg or % Re. 61298056 <LCQ(00119ug/g) 122998002 00663 i 61298057 <toQ(OO119ugig) 12298003 0.0414 i 61298127 61298128 40% 26% * 0 0324 4toQ (0 0302 "gig) 0 0337 C L I X (0 0302 "gig) ctoQ (0 0302 uglg) ctoQ (0 0302 uglg) C t O Q (0 0302 "gig) <toQ(0 0302 "gig) 90298030 0 0344 ctoQ (0 0302 "gig) 12298045 194 I2298044 I8 7 12298043 I8 9 12298042 20 9 18 3 150 12298051 14 1 12298050 17 I 12298049 13 8 12298048 173 'calibration range. estm + CCVs faded. data entered as eslmted **canfumedlow on 90298 ++ Qualitative data only Mtln PFOSA u#g 00391 0 0267 33% 00313 00310 19 2 I5 5 "due Dale EnlerediAnalysI Dale VefiflediAMlyjt Punty EnIeredNenenfled 4113100. 5124100CSH 121121W MMH 12128100hoi (list al22993a) 12126100 LAC CorrectedPFOS LOQs(0.0119&00121 uglg)toinclude sldcomclion facton new are o 01 i n &io 112 "gig LAC n 2 w n i RSD SIdDrv. MYMSDRPD NA NA 44% 5 21 0.03163 6 09 OW189 5 I3 0 987 10 7 I 65 Ers-8-7 0 Excel 97 3M Environmental Laboratory Lrr Week 8 Rework TOXml-lwer212-100 XIS Analytical Report: FACT-TOX-001 LRN-U2103 513112Wl Page 253 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance# 6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 Study. Pnoducf NumbsrlJerI Substance) Matru Method/RWlSW" Analytical Equipment System N w n k r Insmen1 SoRuweNenan a t e of ErVdetlonJAnalyst Date of Analysi~/Analy~t Dale of Data ReductioniAnalvsl Sample Data 104 Week Dletar).Carcinogemcily Study x i t h Narmw Range (98 1%) N-Ethyl PerlluamstanesulfOWamldoElbAIlOl SIl Rats T-6316 (ElFOSE-OH) Rat Lwer FACTI-M-I0 & FACT-Y-2 0 reworked using ET%-7 0 Fl1e-e See t a l 10 roght Chick 080697 and Madeline 041098 R . s q d value See Amhmenls MassLym 2 3 and 3 1 Slop See Anachmenls 06101198 RWWIOK ~-intercept See Attachments 06/12198,06114198,06/18198.09102198 HOJNHJLAC 03116/W, 11129100, 121OllW. I21lOiW IASJMMWWH WEEK 8 RAT LIVER C0"L 27 2 PFOSAA Dilution Factor I PFOSAA Cdc. Cont. ws 27 2 Fl1m.m 6189834 Conuo1 0 0 mgrng C90718M C90737M C90756M OW 1 ~ 130 I - _OW_ I 88 6 I 97 6 1 76 0 1 904 1 156 I 118 1 _3 1_ 3 1 I5 6 I OW sW1?06w5 I2 9 6189835 OW 1w1206006 12.3 61298127 21.0 61298128 72 7 6189845 88 1 6189846 149 6189847 117 6189848 3 10 6189849 15.2 6189852 C911MF CY1145F CY1 162F I High Dose 3W mgkg C90972M C90988M c909%M I 74 406 - 195 79 4 512 594 478 480 384 ~ 306 I 1 1 1 IMO IMO loo0 IMO 1MO IMO I70 40.1 192 78 1 505667 581791 473002 473865 379557 298858 6189853 6189854 6189855 6189856 I22998059 I22998058 I22998057 I22998056 122998055 122998066 00781 0 0920 C91394F 272 IMO 268201 I22998065 C91404F 37s IMO 369672 122998064 C91415F C91438F 'OS = PernuomOCmeEUlrOMte - 293 439 IMO IMO 286207 42m3 122998063 122998062 286 429 + 330 20 3 67 2 PFOSA = Perlluomoclanesulfonarmde + CCVs failed data enlered as estimated PFOSAA ~ Per~uoro~tanrrulConamldoacetatr W O S E = Narmw Range N-Ethyl Perflwmocmesulfamtdo ethyl alcohol '*confinned low on 90298 I* Qualitatwe data only Date Fn1eredJAna1yst Date VenfledJAnalysl Punty FnteredNenfied 4/13/00,5/24/00 CSH 121121W MMH 12126/W LAC ComtedPFOSLoQr(OOll9 &00121 ug/g)tolncludertdcorreetionfaelorr new LCQS are n n i in & n 112 "gig LAC 02119ini UnblO\m unknown - EIFOSE - COW. Ws 244 OW - - 46 5 30 3 000 OW OW OW - 3 9 2 OW OW OW - OW OW 892 Ellmame y-1 3::- Facer 90298002 90298003 90298110 90298111 OW 90298019 90298020 90298021 90298022 90298023 OW 90298027 OW 90298028 OW 90298029 OW 90298030 OW 90298031 10 OW 10 OW 883 90298036 816 - 805 I 10 OW 90298038 90298042 - 347 650 90298045 90298046 Canorntntian of EtFOSE "USor % RII. (0 0298 "gig) t i <aOW ti (0 0298 "gig) t i 25% CLOQ (0 0298 "gig) t i <LoQ(0 0298 ugig) t i <LoQ(0 0298 "gig) t i < L c Q (0 0298 "g/g) t i 0 0388 CLoQ (0 0298 "gig) t i <LCQ (0 0298 "gig) ++ <LCQ (0 0298 u#g) ++ <LCQ (0 0298 ugig) ++ <LOQ(OO298ugig) ++ OW ++ OW ++ 0883 ++ OW ++ 0807 ++ 0786 ++ OW ++ OW ++ 0339 ++ 0636 ++ MLP" EIFOSE ws <toQ 31% RSD SU. Dlv. MYMSD RPD NA 42% I 00316 0 wM2 <top I 0338 NA NA 137 0 463 0352 102 0 359 ETS-8-7 0 Excel 97 3M Environmental Laboratory Lm Week 8 Reuork T O X m l - l n e r 2 1 2 - I 0 0 xls Page 254 3M Medical Department Study: T6316.1 AMDW 092597.1 Covancd 6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 . _.. T-6316 (UFOSE-OH) Rat LlV.3 FACTM-I 0 Br FACT-M-2 0 ~ r o r k r du m g ETS-8-7 0 Chxk080697and Madelm 041098 MssrL)nr 2 3 and 3 I 06101198 RWWIOK 06112198,06114198,06118198.09101/98HOJWWWLAC 031161W, Ill29IW, I2101I00, 12llOlW IASiMMHJKJH Fileme R-SquaredValue slop Y-lnlerrepl See Amchmenlr See AtIaChmeNS See A t ! x h t t S See A n a c h l s WEEK8 R &UP Daw LIVER REWORK Sample # HZO BL-2 Rabbit h y e r BL-1 Rabbit Lver Blk-2 C5071N-MSD C90717h4 I C90718M c90737M C90756M CW769M C91126F I C91140F C91145F C91162F C91163F C90971M C90972M C90988M C909%M C91017M C91393F C91394F C91404F C91415F C91438F tanesulfomte ~toQ(oollou$ig) -1773 2776 <toQ(0 0110 "gig) <LOG(0 I I2 "pig) -1331% I 2 78 2731 I144 245 346 511 1094 I 1250280 1111564 1249941 890777 830679 873644 945819 1027570 1054710 1423345 0 245 0 346 0511 109 0 552 1112 874 946 1028 1055 1423 Date Enteredlhlysf Date V e n f i e d i h l y l t Punty ENeredNenfied 4/13/00, 5124IWCSH 12112100MMH 12/28/00 ha, (list a122998a) 12126100 LAC Comeled PFOS LCQr (0 0 I 19 Br 0 012 I upig) 10 includp 51d correction fxtors new L O Q s are 0 0110 Br 0 I 1 2 "pig LAC 02119101 <Lop <LOQ .l410% 0 549 I 1067 1065 NA NA 11% 59 8 0 328 I85 197 20 0 213 66 3 0.w 41 4 48 4 30 8 32.4 ctoQ (0 0302 ugg) 33 7 00663 i ~LCQ(OO119ug/g) 00414 i 40% *' 26% *' 0 0324 <top(0 0302 Udg) 00337 - Men" - PFOSA "Yg - 0 0391 - 0 0267 - 33% <toQ(00302 "pig) CtoQ (0 0302 UdR) 34.4 <LCQ (0 0302ugrg) 19353 <toQ(0 0302 udg) (0 0302 U d g ) 0 0344 4.W (U 0302 "gig) 194 _0.03_13 - 00310 I 20877 I 209 18323 I8 3 14999 I5 0 14071 14 1 17148 17 I 13844 I3 8 17272 173 * Dah above the Imar cahhation range,e r t m t e d + CCVs bled, dm enteredas estimated "confirmed low on 50298 ++ Qualitalrve dabonly - 19 2 - I5 5 44% 5 21 0 W163 6 09 OW189 10 7 I 65 3 10 I5 2 I 70 401 78 I 505667 581791 473002 473865 379557 298858 268201 369672 286207 429003 0 117 !a 1 1cLOQ(0633ug/@ cLOQ (0 633 udg) I .. .. <LCQ (0 633 Udg) 0 0980 I 0 192 00781 0 0920 474 380 483 299 268 370 286 429 330 35.8 00351 61 I 00561 15 I 72 7 20 3 67 2 0.M cLcQ(oo298upig) i t 1 !K 1 1 1 388 00388 i t 00316 I OW I <LoQ(00298"pig) ++I I 0 <LoQ (0 0298 U&) ++ 0 W <LCQ (0 0298 upig) ++ 0 W <LCQ (0 0298 ugg) ++ <LCQ 000 OO' Wf 807 0 807 0 338 786 0786 it OW OW tt OW OW ++ 339 0339 it 636 0636 ++ 0352 12 7 000401 NA NA 1137 0 463 102 0 359 ETS-8.70 Excel 97 3M Environmental Laboratory LIT Week 8 Rework TOXaO1-l~ver21?-1GXIS 5'3112001 Page 255 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covancdl6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 StUdY RodwtNumbeQesl Substan-) MakiX MdWwaian h l f l r c a l Equipment SydcmNumber l n J t m t SonwareNenion Date ofE x t r a f l i o d h l y s t Date of A n a l y s d h l y Dale of Dab R e d u c t ~ o a ~ h l y d Sample Data 104 Week B c l a r y Carcinogenicity study with N m u Range (98 1%) X-Fthjl Pcrfluomwrrnesulfonarmdo Elhaool m Rats Td316 (EtFOSEOH) Rat LlVer FACT-M-I 0 & FACT-MI 0 rero&ed using ETS-8-7 0 Chick 080698, Amelm 062498, Madeline 041098, and soup 020199 F k n m Sol List to Right R-Squared Valve Le A l l a e k l s S l a p Le Affaehmnta M a r r L p 2 3 and 3 I Y-lntmepl. Le Atfachmenls 06102/98.01114/99,01/17/99 RWW/IAS/SAH 06/25198,06/26/98, 12/14/98,0lll8/99.02/16/W. 02/18/99 HOIKJHIMEEDRB 04/05/00, 11!3O/W, I M I I W , IMSIW. IL'07/W.O1'OMI CSHIMMILKJH PFOS PFOSA 12/14/98 La1 193 06/26/98 LoIL-2353 01118l99 Lat 171 I1114l98 LofL-2353 02/16/99 La1 171 011'18l99 LofUnLnoun 02'18199 Lo1 171 PFOSAA EtFOSE 06/16/98 Lot617 06/15/98 Lot936 11114198 Lot617 02I18I99 LotUnLnoun 02/18199 LotUnLnoun WEEK 14 RAT 1 &UP Dose Method Elk Malm BL W-IWppb W .250 ppb W-750ppb cmvp I C0"tml OOwh Gmup 1 Low Dou 3 0 mglkg cmvp 3 Mid Dosc 30 0 m a g ER REWORK Sample # HZOBlk-l H2OBIk-2 abbil Liver Blk-I ahbit Liver Blk-2 C90768M-MS Z90768M-MSD C9078OM-MS 390780M-MSD C91174F-MS C91174F-MSD C93731M C90880M C91281F C91288F C91299F C91304F C90905M C90907M C90917M C90921M rWwM C91337F C91355F C91370F C91379F Inlllsl WL L I OMX) 1 OOM) 1 0080 I 0080 IO214 10214 10166 10166 IO123 10123 10133 IOZOl I0210 10173 I0277 10099 10123 10104 10378 10~4 IOlSO I 0150 I 00% Total Mmrs e NA NA 40 13 40 13 NA NA NA NA NA NA NA NA NA NA - NA NA NA NA - NA NA NA NA NA NA NA ~ NA NA NA - NA NA NA NA NA NA NA ~ NA NA NA - NA NA NA NA NA NA - ~ N4 NA NA NA - NA NA PFOS Std cormtian F..tW 0 9275 0 9275 0 9275 0 9275 NA NA NA NA NA NA 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 927s 0 9?75 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 077\ 0 9275 0 9275 0 9275 0 9275 0 9275 SI241W. 11107100. 12/141W I2/28100 HOJ 11126JW LAC La1 171 PFOSPvrify PFOS PFOS Fileanme Comrflou F~lor 08640 I I Corn. ndg OW Dilulian Faelor I 1 M011899013 08640 OW I S021899w3 08640 I OW I I 1 MOll899014 08640 OW I S021899w4 NA 867 I MI21498109 NA 1023 I MI214981 10 NA 889 10 MU21699036 NA 983 10 M021699037 NA I 40 10 so21899015 NA IS1 10 SO21899016 08640 348 4 MOI 1899025 08640 425 4 MOI 1899026 08640 504 4 MOI 1899027 08640 383 4 MOI 1899028 I2W I20 . 08640 286 4 MOI 1899029 901 0 901 I23 08640 133 4 MOI 1899032 417 0417 08640 I55 4 MOI 1899033 489 0 489 08640 175 4 M011899034 555 0 555 08640 766 4 M011899035 243 0 243 08640 970 4 MOI 1899036 303 0 303 0402 08640 230 125 M011899039 22950 23 0 08640 231 0.8640 247 08640 210 08640 266 08640 214 08640 235 08640 289 08640 195 08640 256 08640 263 08640 720 08640 261 125 125 I25 125 125 125 125 125 I25 2wO 2WO 2wO M011899040 M011899041 MOI 1899042 Mol1899043 M011899046 Mol1899047 YO1 1899048 MOI 1899049 MOI 1899050 M011899053 Mol1899054 M011899055 22597 24494 20514 26220 21300 23437 28206 19334 25542 41221 114260 40451 22 6 24 5 20 5 26 2 23 4 21 3 23 4 1236 41 2 1405 08640 !%2 2WO MOll899056 153905 08640 339 2WO MOI 1899057 54251 8; 0x640 872 zwo MOIMYWM 137003 08640 858 2wO M011899061 134654 08640 709 2W0 Mol1899062 113467 08640 148 2wO Mol1899063 233295 233 MOll8990M 84401 84 4 415193 415 543101 543 0 8640 M021699024 372033 372 0 8640 314243 3l@!* 262314 296535 248810 0 8640 MO21699032 649710 0 8640 ID00 M021699033 216709 Note PFOS dab may b b1-d bgh h on Ibe MSIM: + Brackelcdby a CCV ill-30% differace,&la may be b i d low LAC 02/19/01 * Reportd below the LCQ ++ PFOSAA MSihlSD mot N l y b-kefed LAC 12I14IW i++ ElFOSE MSiUSD aported,only a 4 p i n t EWF could be eenerated LAC I Zl141W E = Sample went to d q c s s aRer the initial analyrin No extract remaining for m l y s a 'iPUalilative &la only. h c d on MSMSD r e d i s 21 4 0262 32 I 0 129 19 I6 2 I4 ?4: 63 0 398 56 0 UnLnoUn UnLn0V.n Unlvuull UnlvuUll UnLnOWn UnLnOWn UnLnOWn UnLnOUn UnLnowo UnLnoUn UnLnOUn UnLnOWn Ul!kllOUn llnknnun 2.74 I 2 68 36 5 I 35 9 660 I 6 43 9 75 I 960 29 9 I 296 2 44 I 2 41 OW I OW 133 10 1319 111 10 I084 89.7 10 889 134 100 1305 98 3 IO 968 I94 I927 171 ,607 158 153 IW I77 127 1W 12659 IM 1W 15721 142 1W 14058 143 IW 14066 167 1W 16522 Corrected PFOS LCQs (0 0619 & 0 0309 =gig) lo include rld camclian factors ~ ~ w L ~ a r r 0 0 4 9 6 & 0 0 2 4 8 u g L1AgC O t l P / O l "dg o r x kc. MSJMSD RPD N. .A. . .. NA NA N. .A. I N...A. NA NA MI21498014 00537 MI21498015 < C Q ( O O l ? l u#g) 76% NA I I NA MI21498016 < O q ( O O l Z l u&) MI21498017 00359 ~ 1 2 1 4 9 8 0 2 1a o Q ( o o 1 2 1ug/g) a w ~ 1 2 1 4 9 8 0 2 2 (o 0121 "de) 1 MI21498023 002% MI21498024 <GQ(OOl2l uglg) MI21498025 UCQ(OOl2l up'g) MI21498060 I 132 I {f MI21498061 MI21498062 MI21498063 MI21498064 MI21498068 0968 I93 Ml,,d(lsN.O I," 00284 I I I 00156 I I I I I1 13% NA NA 71 I 00202 50 2 OW784 175 0 194 24 2 M062698076 M062698077 MI21498100 127 M121498101 157 MI21498102 14 I MI21498103 14 I MI21498104 165 10 4 14 6 I52 ~ ~ s - 8a- 7 3M EEnx-v197ironmental Laboratory Page 256 3M Medical Department Study: T6316.1 AMDT# 092597.1 CovanceU632C212 SfUdY Roduel N u m W e o t Subotanse) M*hix MeUlod/Re"lSion Andylical Equiuipmenl System Number Irutrumc"l SonralxNemlon Dale af ExlractiodAnalyd: Date of Analysir/Analy.l Dale ofData RedustiodAnalysf Sample Data 104 Week Dietary Cmmogenicify Study ubth N m w Range (98 I%) N-Ethyl Pernuo-tanerulfonarmdo T-6316 (EtFOSEOH) Elhano1 in Ram Ral Liver Film FACT-M-I 0 & FACT-Mf 0 Chick 080698, h l i a 062498. Madellne 041098, and Soup 02olw RSqusvFd Valve Slope MasrLynr 2 3 and 3 I 5111/98,6/298,01114/99 RWWIIASISAH Y-lnlorcepl 6/5/98, 12/14/98.01/18/99,02/16/99,02/18/99 HOJXIIbMEWDRB 12/22/98, 01/20/99,0922199, 3/23/W HOJrWlMRBihLMH See Anashmenls See Anashmenls See Anashments See Altashmeofs Lo1 617 or UnLnawn H20 Bk-l unknown Lo1 936 or Uolroown - EtFOSE Con*. *OW H2O Blk-2 NA ~ C90768M-MS C90768M-MSD C90780M-MS C90780M-MSD C91174F-MS C91174F-MSD C90731M NA NA NA NA unknown NA NA 905 1315 OW - NA NA _NA_ NA - _NA_ 516 480 OW C90746M OW C90748M OW C90768M C90780M C91129F OW - O W OW C91155F OW C9116OF OW C91174F C91181F 21 8 C90797M - OW OW NA C90807M 145 NA I NA NA CW818M 180 I0 MI21498062 1786 I 79 Uhown NA I NA NA C90831M C93836M C91192F 203 I0 MI21498063 1984 198 175 10 MI21498064 1719 I 72 218 10 MI21498068 1168 2 17 18 9 185 0351 Uhoun NA I NA NA UnLnOwn NA I NA NA ~ UhOUn NA I NA NA C91223F 10 MI21498069 3240 3.24 UnLnOWn NA I NA NA C91233F 262 10 MI11498070 2556 2 56 UhOWn NA I NA NA C91241F C91248F C90843M 232 10 MI21498071 2295 229 17.4 303 10 MI21498072 3023 3 02 266 0 462 7 58 IW MU62698067 142 0742 * - U h O U n NA tihown NA UnLnOUn 234 I NA NA I NA NA I A062598076 229 C90852M 202 1W M062698068 20013 20 0 UhOW Ill I A062598077 175 C90863M 1W M062698069 11226 I12 UhOWn 325 I A062598078 314 C90877M 358 1W M062698070 3J70b 35 7 72 8 UhOUn 146 I A062598079 146 C90880M 168 ~ I A"n,sQn"a" 367 C9l18lF 152 I A062598083 149 C91288F 142 I A062598084 139 C91293F UhOWn 171 I A062598085 171 C91299F C91304F C90905M - M U 117 1 A062598086 115 170 1 A062598087 165 943 UhOW NA 1 NA NA C90907M 749 14 0 UhUn NA C90917M 619 61 2 UnlmoUll NA C90921M C9019MM C91329F C9133lF 1209 IW MI214981W IC0 M121498101 , 40356 78625 1164 (07 40 4 78 6 , __ 29 7 UnkoOUn NA , /PI >:.A UhOWl NA NA Uhown NA I NA NA C9135SF IW MI21498102 63619 63 6 UhUn NA I NA NA C91370F C91379F IW MI21498103 50197 50 2 24 6 640 1W MI21498104 63386 63 4 59 1 14 6 Nole PFOS dnfa m y be b l e d high bssedathe MSMSD m v e n e s UhVn UhOwO - NA I NA I NA NA NA NA PFOS = Pcrfluamoclanenulfonsl~ PFOSA = Perflw-tancrulfa&& PFOSAA = Pernuomoctan~sulfonarmdoacct.tc + Bmckeld by I( CCV at -30% diNaocp, dab may be b i e d low LAC 0219m1 * R s p r i d below the LCQ ** PFOSAA YSMSD n ~ful lly braekeled LAC I 2 I 4 I W EIFOSE = N m u Range N-Ethyl Perfluomoctanesvlfondo ethyl alwhol Date EnteredIAna1yrt. 512402, I M 7 / W , I2114JW CSHKJHILAC + i t EIFOSE MSIMSD reponed,only a 4 pi01EWF wuld be gencratcd LAC I2/14/W E = samplew n ~to +*s an*. the *nitidrnalyslr xa for -lY.i. U Qualilalive data only, b a d on M W S D m d l n Dale verifidhlpt 1228lW HOJ k t y EnleredNmfied 1216IW LAC Comcted PFOS LCQs (0 0619 & 0 0309 uglg)to include rld wmlillan fsclorr oew L C Q a . 0 04% 8- 0 0148 ue'g LAC 01'19IOl Analytical Report: FACT-TOX-001 LRN-U2103 NA x NA x NA x NA x NA x NA x NA u NA x NA x 0 229 U 0 175 U 0 314 U 0 " 146 !L7 # * 0 149 # 0 139 # 0 171 u 0 115 # 0 165 # NA # NA NA ? 2w 0 148 NA x NA . NA NA x NA NA NA NA NA 32 8 "5757 IS I 0 0224 NA NA ETS-8-7 0 3M EEnxevl9i1ronmental Laboratory Lw Week 14 Rework TOXMll-lwer212-I0X0 IS Page 257 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covanee# 6329-212 Study Rodwl N u m W e R Substance) Malm MclhdRcuision h a l f l i d Equipmnl System Nvmbr Innrumen1 SoRwarrNenion- Date O f D i W n m d h a l y s t Date Of AonlyndAoalp. Date ofDataR~dwt~odAnalysl Sample Data 104 Week betar~y~ i n o g e n e es~tuidyy with xmu,hg(9C 8 1%) N-Ethyl P d l ~ m t m ~ l S o o m dEtohanol in Rats 1 4 3 1 6 (EtFOSEUH) Rat Liver F,*W S e LtR lo Rtghl FACT-M-I 0 & FACT-M-2 0 m w d e d wmg ETS-8-7 0 R-Squand Val- S e Altaehmnls Chiok 080698, h l l a 062498, Madelme 041098. and Soup 020199 Mar%LLyrU23and31 slope Y-ln(empt See AttaehmnN See Atfsbmmts 06/02/98,01/14199,01/17/99 RWIIASISAH 06/25/98,06/26/98, 12114198.01/18/99.02/16/99,02/18/99 H O 1 ~ ~ W D F . E 04/05/00. 11l30~001, 2/01100, Iz/o5iw. 12/07/00, 01/02/01 CSHIUMWKJH WEEK14 RA' IVER REWORK Group Dor Melhd Bk s.mp1. I H2O BIk-l H2O Elk-2 PFOS Cdc. COOC. os/r 000 000 conmtmtioo olPFOSA vgtg w Y. Rs. LOQ IO 00604"gig) NA Mafm Bk QC-IWwb Qc - 250 ppb Q c . 7 5 0 ppb labbil Liver BIL-l labbil Liver Bk-2 C90768M-MS C90768M-MSD C90780M-MS C90780M-MSD C91174F-MS C91174F-MSD 0.00 0.00 -351 -199 -27 65 1136 I245 LOQI0 wM4 'gig) NA 81% 71% 76% NA NA NA NA NA 13% 665 NA 1070 croup 1 C0"rnl 0 0 mukg C90731M C90746M C90748M C90768M C90780M C91129F C91155F 1100 1332 1603 I 2w WI 417 489 00537 n O Q ( o o l 2 l 'gig) E ;LOQ(00121 "gig) 00359 n c Q ( o O l 2 l "@E) :LCQ(00121 "@E) 0 0284 172 140 E 71 1 206 0 0202 28 5 74 5 93 4 C9llMF 555 C91174F 243 C91181F 303 002% rLCq(ool2l "gig) r L ~ ( o O l 2 "l gig) 00156 68 4 50 2 229 0 00784 11 3 C907YIM CW807M 22950 12597 132 2363 I08 1414 C90818M 24494 0 889 I786 C90831M C90835M c91192F C91223F 20514 26220 21300 23437 I31 17 5 I984 0 %8 I I1 0 194 1719 I 93 2168 2 17 I70 3240 3 24 C91233F 28206 I79 2556 2 56 Group 3 Mid%= 30 0 mgkg C91241F C91248F CW843M CW85ZM C90863M C90877M C9088OM ~91281~ C91288F C91293F C91259F 19334 25542 41221 114260 40451 I53905 54251 1'1003 134654 113467 233295 137 ~ 135 * 113 + 233 + -$ I78 I82 6 67 9 04 563 10 I 9 84- 8589 8 59 6480 6 48 7359 7 36 39 8 8935 894 4 61 2295 2 29 1 80 00831 3023 3 02 742 0742 20013 20 0 11226 I12 35 7 32557 5269 13748 12775 2%78 29 7 C91304F 84401 84 + 141 56 0 8512 8 51 13073 13 I C90905M 415193 415 18099 I8 I 94 3 C90917M C90921M C9096oM C91355F C91370F 543101 543 15579 IS 6 TJ-JrL 372033 372 314243 314 316914 262 248810 I5150 I5 2 24 0 16630 166 392 17258 17 3 12659 I2 7 17?1 I5 7 14058 14 I 649710 I4 I 74 0 61232 61 2 7 28 116449 116 '0 7 40356 40 4 78625 78 6 63619 63 6 50197 50 2 16521 16 5 14 6 152 63386 63 4 Note. PFOS & my b biased lugh ba=c the MSMSD rccovmes EtFOSE = N m w Range I\'-Ethyl Perfluo-tanesulSomonarmdo ethyl nlwhol + Bmcketcd b! * Repand b l a +* PFOSAA h -CV at -10% diNemnEe, .I- M.- 3 may b biased law LAC 0?/19/01 ZSDnolfullyhcketed. LAC 12/14/00 +** EIFOSE h G D rrponed, only a 4 p i n 1c w o could b gemeraled LAC IY14IW E = Sample wnt to drymessaRer the initial analysis No exlml m&g far d y n i s 5/24/00, 1>07/00, I2/14/lN CS-C R Qualilslivc data only. basd on MSMSD msulfs. lU28iW HOI 12/26/00 LAC Corrected PFOS LCQs (0 0619 & 0 0309 udg) lo include rld c o r n c l l ~ nfaclorr new LOQEare 0 04% & 0 0248 uglg LAC OY19iOI Analytical Report: FACT-TOX-001 LRN-U2103 174 2 66 72 8 MO I4 9 8 95 I!I __29 11 . 7 3 24 6 59 2 14 6 M"n RSD EIFOSE Std DN ude x M S M S D RPD I UoQ NA # d UOQ NA # # NA NA # X NA NA +++ b++ 547% 7% n # # # NA P UOQ NA # # # 1 NA # cop # NA I) NA d NA # tJA NA # NA NA NA # NA # NA # NA a NA NA X NA NA 229 0 229 # 0.175 x 0314 # 146 0 " 146 !17 # 0 149 0 139 0 171 0 115 15.1 165 0 165 0 0224 NA NA NA NA NA NA N::.A NA NA ...N A 1VA NA NA NA NA NA NA NA NA NA NA NA UA ETS-8-7 0 3M EEnxveli97ronmental Laboratory Page 258 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance# 6329-212 Study: Product Number(Test Substance): Matrix: MethodRevision: Analytical Equipment System Number: Instrument SohareNersion: Date of Extractiodhalyst: Date of Analysis/Analyst: Date of Data ReductiodAnalyst: Sample Data 104 Week Dietary CarcinogenicityStudy with Narrow Range (98.1%) N-Ethyl PerfluorooctanesulfonamidoEthanol in Rats T-6316 (EtFOSE-OH) Rat Liver ETS-8-6.0 & ETS-8-7.0 Filename: See Below Davey 070799 R-Squared Value: See Attachments MassLynx 3.3, 3.4 Slope: See Attachments 2/29/00 SAL Y-Intercept: See Attachments 05/08/00,05/24/00, 08/2 1/00. 08/23/00 IAS/HOJ/MMH 05/09/00,05/25/00, 08/22/00, 08/28/00 IASIMMHIHOJ Analytical Report: FACT-TOX-001 LRN-U2103 WEEK 53 RAT LIVER Group Dose Sample # Method BU RBL02290-HZO Blk-5 Matrix B L RBL02290-HZO Blk-6 FEiL02290-Liver Blk-5 QC - 250 ppb RBL02290-Liver BIk-6 C90714M-250 ppb-MS-5-1 C90714M-250 ppb-MS-5-2 Group 1 C90714M Control C90751M 0.0 mgkg C90752M C90754M C90767M Group 1 C91121F Control C91122F 0.0 mgkg C91 l24F C91133F C91152F Group 4 C90902M Low Dose C90918M 3.0 m a g C90950M C9095 1 M C90957M Group 4 Low Dose C91323F C91347F 3.0 m a g C91352F C91367F C91371F FFOS - r"crl;uuruu~-icbuirunai. Initial Wt. g 1.0000 l.Oo00 1.oooO I .0000 1.0093 1.0093 1.0093 1.0033 0.9994 0.9912 0.9845 0.9972 1.0073 1.0079 0.9918 1.0162 0.9925 1.0022 0.9878 1.0037 0.9880 0.9939 0.9917 0.9983 0.9908 0.9903 Lot 171 Total Mass of Liver PFOS Std Correction PFOS Purity Correction Surrogate Verified PFOS Cone. PFOS Dilution PFOS Calc. Couc. Filename g Factor Factor NA 0.9275 0.8640 NA 0.9275 0.8640 40.13 0.9275 0.8640 40.13 0.9275 0.8640 NA NA NA NA NA NA NA 0.9275. 0.8640 NA 0.9275 0.8640 NA 0.9275 0.8640 NA 0.9275 0.8640 NA 0.9275 0.8640 NA 0.9275 0.8640 NA 0.9275 0.8640 NA 0.9275 0.8640 NA 0.9275 0.8640 NA 0.9275 0.8640 NA 0.9275 0.8640 I E I ::zI NA 0.9275 0.8640 : :g. NA 0.9275 0.8640 NA 0.9275 0.8640 nglg Factor NA 2.15 1 NA 0.00 I NA 0.00 1 NA 3.16 I Confmed low 1239 1 Confmed low 1006 I NA 706 1 NA 473 I NA 993 1 NA 857 1 NA 526 1 NA 374 1 NA 383 1 NA 539 1 NA 668 1 NA 474 1 NA 739 1000 I I ;::I I NA 548 1000 E E11 NA IO08 1000 NA 820 1000 riels 1.72 0.00 0.00 2.53 528 298 560 378 796 693 428 30 1 305 429 540 374 596825 437941 923568 gpan! 817501 661438 A0821000 16 A0821000 17 A0821000 18 A0821000 19 DO50800019 DO50800020 D0508DO50800024 DO50800025 DO50800026 DO50800027 DO50800030 DO5080003 1 DO50800032 DO50800033 DO50800034 DO50800037 I DO50800038 DO50800039 nCl5ClaClnn4Cl DO50800041 DO50800044 NA 0.9275 0.8640 NA 1065 1000 860688 DO50800045 NA 0.9275 0.8640 NA 959 1000 770046 DO50800046 NA 0.9275 0.8640 NA 776 1000 627330 DO50800047 NA 0.9275 0.8640 NA 678 1000 548628 Do50800048 ' Hi& recovery was confmed on BiiiiOO, Mawas nor enrered because it was outside the caiibiration range. 4iiOiOi mmh Concentration of PFOS uglg or % Ree. <LOQ (0.00491 uglg) <LOQ (0.00491 uglg) 100% 0.560 0.378 0.796 0.693 0.428 0.301 0.305 0.429 0.540 0.374 597 438 924 770 627 Mean PFOS 138% 0.571 1I I10.390 695 694 RSD Std Dev 56% 30.7 0.175 1:25.5 0.0993 17.7 ETS-8-7.0 3M EEnxcveli9r7onmental Laboratory Lvr Week 53 TOX-001-liver212-1OO.xls 5/31/2001 4:48 PPMage 259 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covanc& 6329-212 Study: ProductNumber(Test Substance): Matrix: MethodiRevision: Analytical Equipment System Number: Instrument SoftwareNersion: Date of ExtractiodAnalyst: Date of AnalysidAnalyst: Date of Data ReductiodAnalyst: Sample Data 104Week Dietary CarcinogenicityStudy with Narrow Range (98.1%) N-Ethyl PerfluorooctanesulfonamidoEthanol in Rats T-6316 (EtFOSE-OH) Rat Liver ETS-8-6.0 & ETS-8-7.0 Filename: See Below Davey 070799 R-Squared Value: See Attachments MassLynx 3.3, 3.4 Slope: See Attachments 2/29/00 SAL Y-Intercept: See Attachments 05/08/00,05/24/00, 08121/00,08/23/00 IASIHOJIMMH 05/09/00,05/25/00, 08/22/00, 08/28/00 IASIMMHIHOJ Analytical Report: FACT-TOX-001 LRN-U2103 WEEK 53 RAT LIVER Lot L-15709 Group Sample #l PFOSA Purity Surrogate PFOSA Dose Correction Verified Conc. Method Blk RBL02290-H20 Bk-5 Factor Unknown np/p NA 0.00 RBLO2290-H20 Bk-6 Unknown NA 0.00 Matrix Blk QC - 250 ppb Group 1 Control 0.0 m a g I RBL02290-Liver Blk-5 RBL02290-Liver Blk-6 I C90714M-250 ppb-MS-5-2 C90714M I C90751M C90752M Unknown Unknown NA Unknown Unknown I NA 2.58 NA 0.00 I I ConfmedLow 288 I NA 1 0.710 NA NA C90754M Unknown NA Group 1 C90767M C91121F Unknown - NA NA Control C91122F Unknown NA 5.41 0.0 m a g C91124F C91133F Unknown Unknown NA 25.5 NA C91152F Unknown NA 9.87 Group 4 Low Dose 3.0 mgikg C90902M C90918M C90950M Unknown Unknown Unknown NA 296 NA 196 NA 289 C90951M 1 lnknnwn NA 716 C90957M Unknown 2nd Analysis OK 404 Group 4 C91323F Unknown NA 236 Low Dose C91347F unknown 2nd Analysis OK 216 3.0 mgikg C91352F Unknown Confmed High 219 C91367F Unknown High, Not Confmed 264 C91371F Unknown High, Not Confmed 3 19 PFOSA Dilution Factor 1 1 1 1 1 I I I I 1 1 1 1 1 1 1 IO0 Inn 100 100 100 PFOSA Calc. Cone. nglg 0.00 0.00 2.58 0.00 230 285 0.703 35.4 11.7 18.3 19.6 12.5 5.37 25.3 15.9 9.71 19579 29280 , 7lAR1 40923 21786 21911 26678 32243 Filename DO50800004 DO50800017 DO50800005 DO50800018 DO50800019 DO50800020 A082100024 A082100025 A082100026 A082100027 A082100028 A082100031 A082 100032 A082100033 A082100034 A082100035 DO52400040 DO52400041 DO52400042 -M <.-7-A .n.n-n-A.7_ DO52400047 DO52400048 DO52400049 DO52400050 A082300016 A0823000 I7 Concentration of PFOSA uelp or % Ref. <LOQ (0.0123 ugig) <LOQ (0.0123 ugig) <LOQ (0.0123 u&) <LOQ (0.0123 udg) 77% 95% 0.0354 0.0117 0.0183 0.0196 0.0125 <LOQ (0.00614 upig) 0.0253 0.0159 0.00971 29.8 2lg9.63 I 1 40.9 21.8 21.9 26.7 32.2 PFOSA <LOQ CLOQ 86% 0.0182 Std Dev MS/MSD RPD 21% I 55.1 0.0100 0.0139 0.00677 28.2 8.44 1 17.3 25.3 4.37 ETS-8-7.0 3M EEnxcveli9r7onmental Laboratory Lw Week 53 TOX-00 1-liver2 12-1OO.xls 513 l/ZOOl 4:48 PPMage 260 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covancetf 6329-212 Study: ProductNumber(Test Substance): Matrix: MethodRevision: Analytical Equipment System Number: Instrument SofhvareNersion: Date of Extractiodhalyst: Date of AnalysisiAnalyst: Date of Data Reduction/Analyst: Sample Data 104 Week Dietary CarcinogenicityStudy with Narrow Range (98.1%) N-Ethyl PeriluorooctanesulfonamidoEthanol in Rats T-63 16 (EtFOSE-OH) Rat Liver ETS-8-6.0 & ETS-8-7.0 Filename: See Below Davey 070799 R-Squared Value: See Attachments MassLynx 3.3, 3.4 Slope: See Attachments 2/29/00 SAL Y-Intercept: See Attachments 05/08/00,05/24/00, 08121/00,08/23/00lAS/HOI/MMH 05/09/00,05/25/00,08/22/00,08/28/00IASlMMWHOJ WEEK 53 RAT LIVER Group Dose Sample # Method Blk Matrix Blk QC - 250 ppb Group 1 Control 0.0 mgkg Group 1 Control 0.0 mgkg Group 4 Low Dose 3.0 mgkg Group 4 Low Dose 3.0 m a g RBL02290-HZO Blk-5 RBL02290-HZO Blk-6 RBL02290-Liver Blk-5 RBLO2290-Liver Blk-6 C90714M-250ppb-MS-5-1 C90714M-250 pph-MS-5-2 C90714M C90751M C90752M C90754M C90767M C91122F C91124F C91133F C91152F C90918M C90950M C90951M C90957M I C91347F C91352F C91367F C91371F LotT-7121.1 PFOSAA Purity Correction Factor Unknown unknown Unknown Unknown NA NA unknown Unknown Unknown Unknown unknown Unknown Unknown Unknown Unknown Unknown Unknown Unknown Unknown Unknown Unknown PFOSA = Perfluorooctanesulfonamide PFOSAA = Perfluorooctanesulfouamidoacetate EtFOSE =Narrow Range N-Ethyl Perfluorooctanesulfonamidoethyl alcohol M556 = CSFI7SOZN((H)CH2C00) PFOSEA = Perfuorooctanesulfonyl ethylamide Date Enteredihalyst: 05/15/00,06/12/00,09/03/00CSWLAC Date Verifiedihalyst: 06/05/00.06/06/00CSH: 02/26/01hoj Purity EuteredNerified: 02/16/01 LAC Corrected PFOS LOQ (0.0613 ug/g) to include std correction factors new LOQ is 0.0491 ug/g. LAC 02/19/01 Surrogate Verified NA NA NA NA Confmned Low Confmed Low NA NA NA NA NA NA NA NA NA NA NA NA NA PFOSAA Cone. np/g 0.00 0.00 0.00 0.00 339 370 8.54 48.7 8.27 44.0 4.96 8.95 4.04 61.6 25.5 16.0 803 402 677 PFOSAA Dilution Factor 1 1 I 1 1 1 1 1 1 1 1 I I 1 1 1 100 100 100 PFOSAA Calc. Cone. n%g 0.00 0.00 0.00 0.00 327 359 8.46 48.6 8.27 44.4 5.04 8.98 4.01 61.1 25.7 15.7 80884 40068 68551 7RA75 60700 49362 41 192 45161 46121 56329 Filename DO50800004 DO50800017 DO50800005 W50800018 DO508000 19 DO50800020 DO50800023 DO50800024 DO50800025 DO50800026 DO50800027 DO50800030 DO50800031 DO50800032 DO50800033 DO50800034 DO5080005 1 DO50800052 DO50800053 nnOE"M54 DO50800055 DO50800058 DO50800059 DO50800060 DO5080006 I DO50800062 Analytical Report: FACT-TOX-001 LRN-U2103 Concentratiou of PFOSAA ude or % Rec. <LOQ (0.0307 -ugl-g,) <LOQ (0.0307 ug/g) <LOQ (0.0307 uglg) 109% 120% <LOQ (0.0307ug/g) 0.0486 <LOQ (0.0307 uglg) 0.0444 <LOQ (0.0307 uglg) <LOQ (0.0307 uglg) <LOQ (0.0307 uglg) <LOQ (0.0307 ug/g) <LOQ (0.0307ug/g) <LOQ (0.0307 ug/g) 80.9 40.1 68.6 1P z _".< 60.7 49.4 41.2 45.2 46. I 56.3 PFOSAA 1<LOQ 114% <LOQ 47.6 Std Dev :2 NA 5.67 ETS-8-7.0 3M EEnxvceilr9o7 nmental Laboratory Lw Week 53 TOX-001-liver212-I00.xls 513 112001 4:48 PPMage 261 3M Medical Department Study: T6316.1 Study: Product Number(Test Substance): Matrix MethodlRevision: Analytical Equipment System Number: Instrument SofhvareNersion: Date of ExtractiodAnalyst: Date of Analysis/Analyst: Date of Data ReductiodAnalyst: Sample Data WEEK 53 RAT LIVER Group "'"."I AMDT# 092597.1 Covance#f 6329-212 104 Week Dietary Carcinogenicity Study with Narrow Range (98.1%) N-Ethyl PerfluorooctanesulfonamidoEthanol in Rats T-6316 (EtFOSE-OH) Rat Liver ETS-8-6.0 & ETS-8-7.0 Filename: See Below Davey 070799 R-Squared Value: See Attacbments MassLynx 3.3,3.4 Slope: See Attachments 2/29/00 SAL Y-Intercept: See Attacbments 05/08/00, 05/24/00, 08121/00,08/23/00 IASiHOliMMH 05/09/00, 05/25/00,08/22/00,08/28/00 IASMMWHOJ Analytical Report: FACT-TOX-001 LRN-U2103 lot Unhown Conhol C90754M Control C90767M C91121F C91122F Unknown NA NR 1 Unknown NA NR 1 Unknown NA NR 1 C91124F C91133F C91152F Unknown NA NR 1 Unknown NA NR I Unknown NA NR 1 Low Dose 3.0 m a g Group 4 Low Dose PFOS = ?eflAna:a-act-esc C90902M C90918M C90950M C90951M C90957M C91323F C91347F C91352F C91367F C91371F unknown NA NR 1 Unknown NA NR 1 Unknown NA NR 1 Unknown NA NR I Unknown NA NR I Unknown NA NR 1 unknown NA NR 1 Unknown NA NR 1 Unknown NA NR 1 Unknown NA NR 1 PFOSA = Perfluorooctanesulfonamide PFOSAA = Perfluorooctanesulfonamidoacetate EtFOS = Narrow Range N-Ethyl Perfluorooctanesulfonamidoethyl alcohol M556 = C8F17S02N((H)CH2C00) PFOSEA = Perfuorooctane sulfonyl ethylamide Date EnteredlAnalyst: 05/15/00, 06/12/00, 09/03/00 CSWLAC Date VerifieUAnalyst: 06/05/00. 06/06/00 CSH: 02/26/01 hoj Purity Enterewerified: 02/16/01 LAC Corrected PFOS LOQ (0.0613 ug/g) to include std correction factors new LOQ is 0.0491 uglg. LAC 02/19/01 NR NA NR NR NA NR NA NR NR NA NR NR NA NR NR NA NR NA NR NA NR NR NA NR NA NR NR NA NR NR NA NR NP Y.4 N?. _k.L 1 A NR NA NR NR NA NR NA NR NR NA NR NR NA NR NR NA NR NA NR NA NR NR NA ETS-8-7.0 3M EnExvceilr9o7 nmental Laboratory Lw Week 53 TOX-001-liver2 12-100.~1s 5/31/2001 4:48 PPMage 262 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance# 6329-212 Study: Product Number(Test Substance): Matrix: MethodIRevision: Analytical Equipment System Number: Instrument S o h a r e N e r s i o n : Date of ExtractiodAnalyst: Date of Analysis/halyst: Date of Data ReductiodAnalyst: Sample Data 104Week Dietary CarcinogenicityStudy with Narrow Range (98. I%) N-Ethyl PerfluorooctanesulfonamidoEthanol in Rats T-6316 (EtFOSE-OH) Rat Liver ETS-8-6.0 & ETS-8-7.0 Filename: See Below Davey 070799 R-Squared VaSee Attachments MassLynx 3.3, 3.4 Slope: See Attachments 2/29/00 SAL Y-Intercept: See Attachments 05/08/00,05/24/00,08/21/00,08/23/00 IASIHOJIMMH 05/09/00,05/25/00.08/22/000, 8/28/00IAS/MMH/HOJ Analytical Report: FACT-TOX-001 LRN-U2103 WEEK 53 RAT LIVER 1 I Group Dose I I Method Blk I II Matrix Blk Lot NBI 13047-80 Sample # I M556Puritv I I I correction- I Factor RBL02290-H20Blk-5 I Unknown I I I I RBL02290-H20Blk-6 R B L O ~ ~ ~ O - ~L ~kV- ~5~ Unknown Unknown I QC - 250 ppb RBLO2290-Liver Blk-6 C90714M-250 ppb-MS-5-1 unknown NA C90714M-250 ppb-MS-5-2 NA Group I I Control 0.0 mgkg - Group 1 C90714M C9075 IM C90752M C90754M C90767M C91121F I 1 Unknown Unhown unknown Unknown Unknown Unknown Control C91122F Unknown 0.0 mgkg C91124F Unknown C91133F unknown C91152F unknown Group 4 C90902M Unknown Low Dose C909 ISM unknown 3.0 mgikg C90950M Unknown C90951M Unknown C90957M Unknown Surroeate Verified NA NA NA NA Confmed Low Confmed Low NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Low Dose 3.0 mgkg C91347F C91352F C91367F Unknown NA unknown NA PFOSA = Perfluorooctanesulfonamide PFOSAA = Perfluorooctanesulfonamidoacetate EtFOSE = Narrow Range N-Ethyl Perfluorooctanesulfonamido ethyl alcohol M556 = C8F 17S02N((H)CH2C00) PFOSEA = Perfuorooctanesulfonyl ethylamide Date EnteredIAnalyst: 05/15/00,06/12/00, 09/03/00 CSHLAC Date VerifiedIAnalyst: 06/05/00.06/06/00 CSH: 02/26/01 hoj Purity Enteremerifred 02/16/01 LAC CorrectedPFOS LOQ (0.0613ugig) to include std correctionfactors new LOQ is 0.0491 ugig. LAC 02/19/01 M556 Cone. O%P 0.00 0.00 0.00 0.00 296 336 0.00 37.4 0.00 29.0 8.62 5.14 0.00 10.9 8.22 7.79 860 591 546 409 522 395 297 292 307 510 Dilution Factor 11 11 1 1 1 1 1 1 1 1 1 1 I 1 1 100 100 100 1 inn , 100 100 Calc. Cone. 0.00 0.00 0.00 294 333 0.00 37.3 0.00 29.3 8.76 5.15 0.00 10.8 8.29 7.67 86649 58978 55321 407 1 0 52862 29927 29261 30992 51513 Filename I DO50800004 DO50800017 I DO50800005 DO50800018 DO50800019 DO50800020 DO50800023 DO50800024 DO50800025 DO50800026 DO50800027 DO50800030 DO50800031 DO50800032 DO50800033 DO50800034 DO50800051 DO50800052 DO50800053 , -nn.s- n.-an.-n-na. DO50800055 DO50800059 DO50800060 DO50800061 DO50800062 Concentration I of M556 ug/gor%~ee. <LOQ (0.0308 ug/g) I <LOQ (0.0308 udg) <LOQ (0.0308 ug/g) <LOQ (0.0308 udg) 98% 112% 0.0373 <LOQ (0.0308 ugig) <LOQ (0.0308 ug/g) ~~ <LOQ (0.0308 udg) I <LOQ (0.0308 udg) <LOQ (0.0308 ugig) <LOQ (0.0308 ugig) 1<LOQ (0.0308ugig) 4"? 52.9 39.7 29.9 29.3 31.0 51.5 Mean M556 Ue/P <LOQ <LOQ 105% <LOQ 58.9 36.3 RSD Std Dev MS/MSD RPD I NA NA NA NA 13% I NA NA I I 1 %; 9.51 ETS-8-7.0 3M EEnxcveil r97onmental Laboratory Lvr Week 53 TOX-00 I-liver2 12- 1OO.xls 5/31/2001 4:48 PPMage 263 3M Medical Department Study: T6316.1 AMDT# 092597.1 CovanceiY 6329-212 Study: Product Nnmber(Test Substance): Matrix: MethoaRevision: Analytical Equipment System Number: Instrument SofhvareiVersion: Date of Extractiodhalyst: Date of Analysisihalyst: Date of Data Reductiodhalyst: Sample Data 104 Week Dietary Carcinogenicity Study with Narrow Range (98.1%) N-Ethyl PerfluorooctanesulfonamidoEthanol in Rats T-63 16 (EtFOSE-OH) Rat Liver ETS-8-6.0 & ETS-8-7.0 Filename: See Below Davey 070799 R-Squared Value: See Attachments MassLynx 3.3, 3.4 Slope: See Attachments 2/29/00 SAL Y-Intercept: See Attachments 05/08/00, 05/24/00,08/21/00,08/23/00 IASIHOJMMH 05/09/00, 05/25/00,08/22/00, 08/28/00 IAS/MMH/HOJ WEEK 53 RAT LIVER lot 529 PFOSEA Purity Correction Factor Unknown Unknown Matrix Blk RBL02290-Liver Blk-5 Unknown RBLO2290-Liver Blk-6 Unknown NA NA Control C9075 IM Unknown Unknown 0.0 m g k g C90752M C90754M Unknown Unknown C90767M Unknown Group 1 Control 0.0 mg/kg C91121F C91122F C91124F C91133F C91152F Unknown Unknown Unknown NA NR 1 Unknown NA NR 1 Unknown NA NR I Group 4 Low Dose 3.0 mg/kg C90902M C90918M C90950M C90951M Unknown NA NR I Unknown NA NR 1 Unknown NA NR 1 Unknown NA NR 1 Unknown NA NR 1 Unknown Unknown Unknown Unknown Unknown PFOSA = Perfluorooctanesulfonamide PFOSAA = Perfluorooctanesulfonamidoacetate EtFOSE = Narrow Range N-Ethyl Perfluorooctanesulfonamidoethyl alcohol M556 = C8F 17S02N((H)CHZCOO) PFOSEA = Perfuorooctane sulfonyl ethylamide Date EntereaAnalyst: 05/15/00, 06/12/00,09/03/00 CSWLAC Date Verifiedihalyst: 06/05/00.06/06/00 C S H 0212610I hoj Purity EnteredNerified: 02/16/01 LAC Corrected PFOS LOQ (0.0613 ug/g) to include std correction factors new LOQ is 0.0491 ug/g. LAC 02/19/01 NR NA NR NA NR NA NR NA NR NA NR NA Yl hlA NR NA Analytical Report: FACT-TOX-001 LRN-U2103 NR NR NA NR NR NA NR NR NR ??!. I,.TA. NR NR NA NR NR NR NR NA NR NR NA ETS-8-7.0 3M EEnxvceilr97onmental Laboratory Lw Week 53 TOX-OOI-liver2 12- 1OO.xls 513 1/2001 4:48 PPMage 264 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance# 6329-212 Study: Product Number(TestSubstance): Matrix: MethodRevision: AnalyticalEquipment System Number: Instrument S o h a r e N e r s i o n : Date of ExtractioniAnalyst: Date of AnalysisiAnalyst: Date of Data ReductiodAnalyst: Sample Data 104 Week Dietary CarcinogenicityStudy with Narrow Range (98.1%) N-Ethyl PerfluorooctanesulfooamidoEthanol in Rats T-6316 (EtFOSE-OH) Rat Liver ETS-8-6.0 & ETS-8-7.0 Filename: See Below Davey 070799 R-Squared See Attachments MassLynx 3.3,3.4 Slope: See Attachments 2/29/00 SAL Y-Intercept See Attachments 05/08/00, 05/24/00,08121/00, 08/23/00 IAS/HOJIMMH 05/09/00, 05/25/00,08/22/00,08/28/00 IASMMWHOJ Analytical Report: FACT-TOX-001 LRN-U2103 WEEK 53 RAT LIVER Group Sample # Concentration Dose of PFOS ug/g or ?'e Rec. Method Blk RBL02290-H20 Blk-5 <LOQ (0.00491 opig) Matrix Blk RBLO2290-H20 Blk-6 RBLO2290-Liver Blk-5 <LOQ (0.00491 upig) <LOQ (0.00491 upig) QC - 250 ppb RBLO2290-Liver Blk-6 C90714M-250 ppb-MSJ-1 C90714M-250 ppb-MS-5-2 <LOQ (0.00491 upig) 177% * 100% Group 1 C90714M 0.560 Control C90751M 0.378 0.0 mg/kg C90752M 0.796 C90754M 0.693 C90767M 0.428 Group 1 C91121F 0.301 Control C91122F 0.305 0.0 mg/kg C91124F 0.429 C91133F 0.540 Low Dose 3.0 mg/kg 1 C91152F C90918M C90950M C90951M 0.374 438 924 699 C90957M 818 Group 4 C91323F 66 1 Low Dose C91347F 86 1 3.0 mg/kg C91352F 770 C91367F 627 C91371F 549 Mean PFOS ug/g <LOQ <LOQ 138% 0.571 10.390 RSD Std Dev MSIMSDRPD NA NA NA NA 56% 30.7 0.175 25.5 0.0993 Concentration of PFOSA ug/gor%Rec. <LOQ (0.0123 udg) CLOQ (0.0123 udg) <LOQ (0.0123 ugig) <LOQ (0.0123 odg) 77% 95% <LOQ (0.00614 udg) 0.0354 0.0117 0.0183 0.0196 II 0.0125 <LOQ (0.00614 u d g ) 0.0253 0.0159 0.00971 695 I89 40.9 23.7 21.8 21.9 17.7 26.7 694 123 32.2 Mean PFOSA ug/g <LOQ <LOQ 86% 0.0182 0.0139 28.2 RSD Std Dev MS/MSD RPD NA NA NA NA 21% Concentration of PFOSAA ng/g or YORec. cLOQ (0.0307 udg) <LOQ (0.0307 udg) <LOQ (0.0307 ugip) <LOQ (0.0307 udg) 109% 120% <LOQ (0.0307 ugip) 0.05 <LOQ (0.0307 udg) Mean PFOSAA RSD Std Dev +114% 5 5 . 7 1 :: 4.37 56.3 47.6 I 11.9 5.67 ETS-8-7.0 3M EEnxvceilr9o7 nmental Laboratory LVTWeek 53 TOX-001-liver212-1OO.xls 513 112001 4:48 PPMage 265 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance##6329-212 Study: Product Number(Test Substance): Mat& MethodRevision: Analytical Equipment System Number: Insbument SoftwareNersion: Date of ExtractiodAnalyst: Date of Analysis/Analyst: Date of Data ReductionIAnalyst: Sample Data 104 Week Dietary CarcinogenicityStudywith Narrow Range (98.1%) N-Ethyl PerfluorooctanesulfonamidoEthanol in Rats T-6316 (EtFOSE-OH) Rat Liver ETS-8-6.0 & ETS-8-7.0 Filename: See Below Davey 070799 R-Squared Value: See Attachments MassLynx 3.3,3.4 Slope: See Attachments 2/29/00 SAL Y-Intercept: See Attachments 05/08/00,05/24/00,08/21/00,08/23/00 IASIHOJIMMH 05/09/00,05/25/00,08/22/00,08/28/00 IASIMMHMOJ WEEK 53 RAT LIVER Group Dose Sample # Method Blk Matrix Blk QC - 250 ppb Group 1 Control 0.0 mgikg Group I Control 0.0 mgikg RBL02290-H20 Blk-5 RBL02290-H2O Blk-6 RBL02290-Liver Blk-5 RBL02290-Liver Blk-6 C90714M-250 ppb-MS-5-1 C907l4M-250 ppb-MS-5-2 C90714M C90751M C90752M C90754M C90767M I C91121F C91122F C91124F C91133F Group 4 Low Dose C90902M C90918M C90950M C9095 IM Low Dose 3.0 mgikg C91347F C91352F C91367F Concentration of EtFOSE n& or % Rec. NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR PFOSA = Periluorooctanesulfonamide PFOSAA = Perfluorooctanesulfonamidoacetate EtFOSE = Narrow Range N-Ethyl Perfluorooctanesu~fouamidoethyl alcohol M556 = CSF17SO2N((H)CH2COO) PFOSEA = Perfuorooctanesulfonyl ethylamide Date EntereUAnalyst: 05/15/00, 06/12/00,09/03/00CSWLAC Date VerifiediAnalyst: 06/05/00.06/06/00CSH: 02/26/01hoj Purity EnteredWerified 02/16/01 LAC Corrected PFOS LOQ (0.0613ug/g) to include std correction factors new LOQ is 0.0491 ug/g. LAC 02/19/01 EtFOSE NR I NR Std Dev NA NA NA Concentration of M556 <LOQ (0.0308 udg) <LOQ (0.0308 udg) 98% 112% <LOQ (0.0308 udg) I 0.0373 <LOQ (0.0308 udg) 1 i L 0 Q (0.0308 udg) CLOQ (0.0308 ug/g) <LOQ (0.0308 udg) <LOQ (0.0308 upig) <LOQ (0.0308 udg) <LOQ (0.0308 udg) <LOQ (0.0308 udg) 59.0 105% <LOQ RSD Std Dev MS/MSD RPD NA NA NA NA 13% NA NA NA NA 26.2 9.5 I Analytical Report: FACT-TOX-001 LRN-U2103 Concentration of PFOSEA ug/g or % Rec. NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR ?!P, NR NR NR NR NR NR Mean PFOSEA ug/g NR NR NR NR NR RSD Std Dev MSMSDRPD NA NA NA NA NA NA NA ETS-8-7.0 3M EEnxvceilr97onmental Laboratory Lvr Week 53 TOX-001-liver212-1OO.xls 513 1/2001 4:48PPMage 266 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 3M Environmental Laboratory Page 267 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 3M Environmental Laboratory Page 268 3M Medical Department Study: T6316.1 I MI' 8 0o"nhr 3M Environmental Laboratory Analytical Report: FACT-TOX-001 LRN-U2103 Page 269 3M Medical Department Study: T6316.1 AMDTX 092597.1 Cov.rrr# 6329.212 Analytical Report: FACT-TOX-001 LRN-U2103 u...s coo=&= 3M Environmental Laboratory Page 270 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 3M Environmental Laboratory Page 271 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covanee# 6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 IR sample x C90842M C90844M C90850M C90851M C90854M C9085SM C90856M C90858M C908MM C90864M C9086SM C90867M C90869M C90871M C90881M C9088zM C90883M C90884M C90885M C90889M C90893M C909WM C91252F C91253F C91254F C91256T C91264F C91178F C91279F C91286F C91292F C91294F C91295F C91301F C91303F C91307F C91310F inat= 1aitl.l Wt. I 0 9962 I OIM o 9930 1 0565 0 9872 0 %?I I0485 I 0156 I 0370 I0137 I oms n 9923 11355 I nsii I 0129 n 9909 0 9926 0 9874 10068 I 0299 I 0178 10372 I1046 I 0730 n 9834 0 9%3 I0678 11455 I nizn I 0412 10162 I W46 1.0159 I 1103 0 9908 10083 I 0132 Tof.1 Ma.. ariiuer D NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA SFOS Sld Comtbo Fartor n 927s 0 9275 n 927s 0.9275 0 9275 0 9275 0 9275 n 927s n 927s n 9275 0 9275 n 927s 0.9275 n 9275 n 927s 0 9275 0 9275 n 9275 0 9275 0 9275 0 9275 n 9275 0 9275 0.9275 0 9275 0 9275 n 9275 0 9275 n 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 0 9275 La1 171 PFOS Purity correetion F..,., n 8640 a 8640 n 8640 0 8640 n 8640 0 8640 0 8640 n 8640 0 8640 o 8Mn 0 8640 0 8Mo n 8640 0 8640 0 8640 o swn n 8640 0 8640 o 8640 0 8640 o 8640 0 8640 0 8640 0 8640 o 8an 0 8640 0 8640 0 8640 n 8640 0 8640 o 8640 0 8640 n 8640 0 8640 0 8640 0 8640 0 8640 S"WOL.l. Verified NA NA NA NA NA NA NA NA Low, Not C o o l i d NA NA NA NA NA NA NA NA Low, Not C o n C i NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Filename R-Squared Valve slope Y-blercept see Below See Attachments Ses Aua~hments See AUachmenta Box 00412. 00413 PFOS Coo*. n& 975 268 287 7% I104 458 106 836 424 774 469 635 298 595 267 492 239 376 642 232 371 165 437 270 533 403 381 643 IW 592 124 294 56 0 235 41 1 141 319 PFOS Dilufiam Fmlor 200 2w 200 2W 2W ZW 2w 200 Iwo 2w 200 2W 200 2W 2w 2w 200 2W ZW 200 200 zw 5W 5W 500 JW 5W 5W 500 5w 5W 5W 500 5W 5w 5W 500 PFOS C d C . cooe. nr/s 156858 42289 46316 120793 179256 73931 ~n303w038 42 3 A030Mw39 46 4 ~0~03mn 121 AO303wO41 179 ~n303wo44 73.9 16276 A0303M445 I6 3 131992 AO303OM46 132 337224 AO8ISwO88 337 122357 ~0303~48 122 74361 A0303WSI 74 4 102548 AO303W52 103 42001 A0303W53 42 0 90688 A0303WS4 907 42229 A03030055 42 2 7%28 A0303W58 79 6 38587 A0303W59 38 6 61036 AO8ISwO91 61 0 1022% A0303Mm) 102 36130 A0303W6l 36 1 58405 26321 158371 100973 2171in 161881 143127 231922 39747 227867 48865 I 17280 22079 84658 166184 56102 Irllm RSD Std DE" MSIMSD RPD 78 I 70 3 54 n 68 j ETS-8-7 0 3M EEnxevl9i7ronmental Laboratory Lur Wuek IO5 (2) TOXM)I -liver212-lCi.x1s Page 272 5131'2WI 4 48 PM 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance# 6329-212 study PmduelNumbcr(Teslsub~aoeE) Matnr MefhMevismo Aoalytioal EquipmentSydemNumber lnslrumnl SaRuareNemdn Date ofErlractiod~aiys1 Date o f h a l y s i d h l y s t : ~ a o rf m~a ~ e d ~ ~ t i ~ d ~ d ~ a Sample Data 104 Week Dietary Csmnogelucity Study ritb X m x Range (98 1%) N-Ethyl Perflwmoftanesulfondo 14316 (EIFOSEGH) Rat Lwer ETS-8-6 0 & ETS-8-7 0 h i i s 062498, Davey 070799 MassLpx 3 3 02/141W,OY16/W RWWISAL 02/zlIW. 01/29/W, 03/01/00,03/03/00,03'041W, 07128100,OS/lS/W MMHiMEEI1AS 01/28/W, 03/01/W, 03103IM). 031OWW. 07/31/00,08/17/W IASMMH Efhanol in Rats WEEK 105 RAT LI\ Group Dor Sample # 101 L-15709 PFOSA Purity corntion surrop.t. Verified Group 3 NA Mid Dose NA 30 0 W g NA NA NA NA NA NA C908MM unLn0un NA CWSMM Uhoun NA C9086SM UhOWn NA C90867M UhOWn NA C90869M UhOWn NA C90871M UnLnOUn NA C90881M UlblOUll NA W88M Un*ooWn NA C90883M UhOUn NA C90884M UnLnOUn High. Not C a n f d C9088SM UnLnOUn NA C90889M UhOWn NA C90893M UhOW NA CWM UoLnOUn NA Crovp 3 C91252F UnLnOUn NA Mid h a e C91X3F UhOWn NA C91254F UhOWn NA C91256F UhOW NA C912-F UnLnOWn NA C91278F UnlawUn NA C91279F UnLnOWn NA C91286F UnLnOWn NA C91292F UhOUll NA C91294F unknoun NA C91295F LinLnOUn NA C91301F UhOWn NA C91303F UhOW NA C91307F UhOUn NA C91310F LlhOUn NA __.. rrUI ~ esriruomocmesur, PFOSA = Perfluomtanem PFOSAA=Pernuoroactam EIFOSE = Narmw Range N M556 =CSF17S02N((H)CE PFOSEA = Perrwmte : w ,nyl elbylami& Date EnlerediAmIyst /l4/00,03/16/W, 03/11/W,O8/l5/W, 09/03/00LAC Dale Venfiedihalyrf 03/09/01 bo, Fuity EnlcrrdRic6fied 01/19/01 LAC PFOSA COW. nplr 366 188 323 361 472 518 435 314 633 478 224 271 313 454 324 371 270 53 6 424 338 354 334 386 387 452 382 334 513 236 497 354 377 166 309 330 297 266 PFOSA Dilulbm F.CfW 0 0 20 20 20 20 0 20 20 0 20 0 20 20 20 20 0 200 20 20 20 0 20 20 0 20 20 20 0 0 20 20 0 0 0 0 20 ~ PFOSA C.i'. coor. oeh 7349 36% 6514 6828 9569 10434 8293 6178 12569 9434 4441 5466 5521 8631 6403 7493 5439 10849 8430 6567 6949 6625 6992 736 9190 7676 6251 8954 4670 9154 69M 1M9 3266 5559 66d 58% 5242 ~~ A030300087 A030300088 A030300089 A030300090 A0303wo93 A030300094 A030300095 A030300096 A030300097 A03OMoIW AO303WIOI A030300102 A030300103 A0303W104 A0303W107 A030300108 A0303W109 A081500091 A0303W110 A0303WI 1I A0303MI 14 AO3U3WI I5 A030300067 A030300068 A030300069 A030300072 A030300073 A030300074 A0303wo66 A030300075 A030300076 A0303Mx)79 A030300080 A0303Mx)81 A03030008Z A030300083 A030300086 conrcotr.uon of PFOSA Ye's or x h. 7 35 3 70 6 51 6 83 9 57 10 4 8 29 6 18 12 6 9 43 444 5 47 5 52 8 63 6 40 7 49 544 10 8 8 43 6 57 6 95 6 62 699 7 21 9 I9 7 68 6 25 8 95 4 67 9 55 6% 7 51 3 27 5 56 666 590 5 24 RSD Sld Dev M S M S D RPD 29 I 2 17 25 4 87, Analytical Report: FACT-TOX-001 LRN-U2103 - P m s A p - cnor. WE 291 354 506 362 180 977 545 471 583 550 241 253 705 620 350 420 295 839 866 802 430 _48_8 282 430 408 533 271 699 239 91 1 351 471 261 248 422 - 356 ?? PFOSAA Dilution Fmrtor 20 20 20 a 1w 0 20 20 20 20 20 20 20 20 20 20 0 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 PFOSAA Cdr. Conc. nglg 5840 6967 10194 6845 36513 19689 10390 9279 11573 108J4 4761 5093 12412 11794 6916 8486 5935 17W2 17198 15569 8459 9694 5114 8W9 8292 I0693 5070 12207 4729 17499 6906 9383 5143 4470 ,,.-8525 7067 Filmam. COn-b.1i00 dPFOSAA uglg or x b. A0303wO87 5 84 A030300088 697 A030300089 IO 2 A0303w090 6 85 A030303341 36 5 A030300094 19.7 A0303wo95 10 4 A0303000% 9.28 A0303W097 I1 6 A0303WlW IO 9 A0303WIOI 4 76 A030300102 J 09 A030300103 12 4 A0303W104 II 8 A0303W101 692 AO303W108 8 49 A0303W109 594 AO815KW92 170 A0303W110 17 2 AO303Wlll IS6 AO303W114 8 46 A030300115 A030300067 A030300068 A0303wo69 AO303CKx312 A030300073 A030300074 A030300066 A030300075 A030300076 A030300079 A030300080 A030300081 A030300082 ."~"~"""", A030300083 ~YI",wo" Mesa PFOSAA Yglg RSD Sld D r v MS4MSD W D W8 6 95 43 6 3 48 ETS-8-7 0 3M EnExvceli9r7 onmental Laboratory L I T W e e l 105 (2) TOXM)I-liver2l2-IW XIS 5/31,2MI 4 48 PPMage 273 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covanceil6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 WEEK 105 RAT L hUP Dose Group 3 Mid Dose 30.0 &g Gr0.p 3 Mid Dose 30 0 mgkg . .Drnr~."."....",..,~.. ~ -1. :R Smnph # C9084zM C90844M C90850M C908SIM CH1854M C908SSM C90856M C90858M C908WM C9086dM C9086SM C90867M C90869M C90871M C90881M C90882M C90883M C9088bM C90885M C90889M C90893M C909wM C912S2F C91253F C91254F C9125bT C91264F C91278F C91279F C91286F C91292F C91294F C9129SF C91301F C91303F C91307F C91310F lot unknown ElFOSE Purity Ca.-tiO. F..tO, Unknoun unknoun UnknOUll UnknOW UnLm*ll unknown Unl;MWI UnknOW UllklWUll UnknOWl UhWI unknoun UnknOUlI lJhU?l UnknOWI Unknown Un*OO*rn UhOWI Ur*naUIl UnknOWn unknown lin*m.w UbWn UnknOUn unknoun UnknOUn UhW UnknO*ll UnknOull UnknOSIl UnknO*ll UnknOUn unknoun unknoun unknoun UnLnOUll UnknO*rn EIFOSE EIFOSE EIFOSE Fileomme Concealratio Mc.n RSD C00.2. Dilution C.k. Coar. ofEtMSE ELFOSE Sld I*v OYl F.Ct0, nS/s M W S D RPD NA 95 0 I 95 3 A072800017 NA 168 1 166 A072800018 NA 39 I 1 39 3 A072800019 NA 167 I IS8 ~n7280~20 NA 39 I I NA 963 I 39 6 A072800021 no ~072800024 NA 34 5 1 319 A012800015 NA 662 I 65 2 AO~Z~OM)~~ NA 8n s I 80 0 ~n72800027 NA 76 3 I 75 2 ~n72nwo28 NA 137 135 A072800031 NA 80 6 1 81 2 A072800032 NA 86 0 I 7s 7 ~0728~033 NA 51 3 I 48 8 ~0728~034 NA 58 8 I 58 n A072800035 NA 153 1 I55 A072800038 NA I8 4 I 18 A072800039 NA 72 4 I 73 3 ~n728w040 NA 219 I 218 ~07?8~041 NA 49 2 I 47 7 A072800042 NA 81 3 I NA OW I NA 84 I I NA I12 I 79 9 AO7t8000IS OW A0728w046 UCQ(00614ugig) n 0883 76 2 ~ ~ 2 8 ~ 0 4 7 o 0762 10s A0728wO48 n 10s 47 I 00415 NA 969 1 985 A072800049 0 098s NA 97.8 I 98 I A072800052 o 0981 NA 111 I 104 A071803353 0 104 NA 147 129 A071800054 0 I29 NA 24 I 1 23 8 AO728000SS <LCQ(OO614ug/g) NA 111 107 A0728wO56 0 107 NA 154 NA 138 151 A0728000S9 0 151 137 ~n728mm n 137 NA 9 69 NA 660 9 54 A072800061 <LOQ (0 0614 "pig) 59 4 ~0728~062 n 05% NA I In NA NA - 89 I 33 1 122 A072800063 0 I22 88 I ~n7280~66 0 0884 33 3 A072800067 CLOQ (0 0614 ug/g) 0 103 ~ 26 9 o 0276 M556 C.k. cooc. NA 345 20 NA 421 20 NA 614 2n NA 797 20 NA 427 2n NA 255 20 NA 590 2n NA 778 20 nE/L 10857 6798 8480 A030300088 A030300089 680 I 848 11631 A030300090 I1 6 16151 ~0303~093 162 8M3 A0303W094 860 4869 A030300095 487 11618 A030300096 II 6 15449 ~030300097 is4 NA 591 2Q NA 399 zn NA 312 20 NA 453 20 NA S45 20 11779 7888 6286 798s 10364 AO~O~WIW 118 A0303WIOI 7.89 ~n3n3win~ 619 A0303W103 79 A0303M104 104 NA 326 20 NA 520 20 NA 264 2n 6427 AO303W107 643 insw AO303W108 105 5327 A0303W109 5 33 NA 729 20 NA 618 20 NA 413 20 NA 393 20 NA 272 20 NA 537 2n NA 455 20 NA 770 20 14760 12272 802s 7724 5398 9729 8473 15661 AO~ISW~? 148 AO303WIIO 123 AO303Wlll 803 A0303WII I 7 72 ~ 0 3 0 3 ~ 1 1 s 540 A030300067 9.73 A0303wo68 847 ~03n300069 157 NA 512 20 NA 3S7 20 10275 6689 ~n303~072 io3 A030303373 669 NA 584 20 NA 226 20 NA 123 2n NA 479 20 NA 517 20 NA isn 20 NA 415 20 NA 620 20 NA 268 20 Ndl zy 2: 10199 4468 IW5I 9421 10286 2961 7474 Its22 5325 ~rnn"r" " ~0303~074 102 A030300066 447 A03030007S I01 AO303WO76 942 ~ 0 3 0 3 ~ 0 7 9 in3 A0303wO80 296 ~n3n3~081 747 .-.-.~""", A030300082 A0303WO83 I.",",yyyDy 12s ,^. 5~ 3 2 "1 RSD Sld I*v M W S D RPD 34 6 3 29 38 8 J 33 ETS-8-7 0 3M EEnxcvel9i7ronmental Laboratory L\T Week 105 (2) TOXdOl-l1\cr112-100 1. 5 3I'ZWI 4 I 8 PPMage 274 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covancdl6329-212 StUdY m u d Sum!m(Teest S u h c e ) Malm MethdRevamn Analfllsal Equipmmt SystemNumber I n s t m o t SoRwmNemion Date ofExtractiodAnaly* Date o f h l y s d A n a l y s 1 Dale of Dals RcducI~on/Analyst Sample Data 104 Week BeIary Carcmopnxlty Study wlh S a m w Range (98 1%) A-Ethyl PerfluomtanenulfooamidoEthanol m Rats T-6316 (ElFOSEOH) Rat Llwr Filename See Below ETS-8-6 0 R ETS-8-7 0 Armlib 062498, Davey 070799 R-Squared ValuSee Allaobmoltr Slop See A t l ~ n t s MassLynr 3 3 Y-Intercept See Attschmenla 02141W. DYl6iW RWWiSAl 02/21/W, 01/29/W,O3/01lW.03103100,03/041W,07128100,08115lW MMIUMEEAAS BorW012,W013 o z 1 2 8 i ~0. 3rni/w,O ~ I O ~ IVW~,I M I W07. 1311~0. 8117m IASIMMH I`EEK 105 RAT L mse ER s.mpi. # Sulrogmtr vlnnd Gmup 3 C90842M NA Mid Dose C90844M NA 30 0 m g k g C90850M NA C90851M NA C90854M NA C9085JM NA C90856M NA C90858M NA C908MM NA C0864M NA C9086SM NA C93867M NA C90869M NA C90871M NA C90881M NA C90882M NA C90883M NA C90884M NA C93885M NA C90889M NA C90893M NA C909WM NA Group 3 C91252F NA Mid -re C912S3F NA 30 0 m a g C91254F NA c91256F NA C91264F NA C91278F NA C91279F NA C91286F NA C91292F NA C91294F NA C9129SF NA C91301F NA C91303F NA C91307F NA C91310F ~ NA ,mate PFOSA = PerfluomodanenulfonarmdP PFOSAA= ~rnuomoctanesvlronarmdoacclate EIFOSE = Narmr Range S-Ethyl PFrflvomactanesulfonamidoethyl alcohol M556 = C 8 F 1 7 S O l N ( ( H ~ H 2 C ~ ) PFOSEA = Perfuomoslane sulfonyl clhylarmdc Date EnferedIAnalyst 03/14/W, 03/16JW, 03117lW. 08/15l00,09103lW LAC Date VcrificlAnalyrt 03109/01hoj Fumy EnteredNsnGed 02119l01 LAC PFOSEA C*"C OYg OW OW OW OW 294 2 53 3 93 160 OW OW OW OW 0 780 364 6 14 OW OW 0 180 3 29 OW 3 43 000 I03 0 230 OW OW 4 13 2 91 OW 3 35 OW 2 55 000 3 18 OW 16 8 OW DilYLiOll I I 1 I I I I I I I I I I I I I I C.lC. Con;r. OW OW 298 OW 0 687 3 46 6.06 OW OW 0 182 3 27 OW 2 54 OW 2 86 OW 16 6 OW Fik..U.e A072800017 A072800018 A0728W19 A072800020 A012800021 A072800024 A072800025 A072800026 A072800027 A072800028 A072800031 A072800032 A0728W33 A072800034 A0728W035 A072800038 A072800039 A072800040 A0728W41 A072800042 A0728WMS A0728CiM46 A072800047 A072800048 A072800049 A072800052 A072800053 A072800054 A0728WOSS A072800056 A072800059 A0728000M A0728ooo61 A072800062 A072800063 A0728W66 A072800067 Con-mim OfPFOSEA Ys/gor%Rec. < L o p (00307 "dg) <CQ (0 0307 "pig) aw UCQ(00307 udg) (o 0307 <LCQ (0 0307 "pig) <W (0 0307 "pig) UW (0 0307 "gig) UCQ(0 0307 udg) C W (0 0307 "pig) a W (00307 "gig) <W (00307 "gig) <La2(0 0307 "gig) C W (0 0307 ngip) aw <W (0 0307 "pig) (0 0307 "gig) <W (0 0307 uglg) aw aCQ(0 0307 uglg) (0 0307 "gig) <LOP (0 0307 `pig) UCQ(00307 "pig) <W (0 0307 uglg) <CQ (0 0307 "gig) a C Q (0 0307 "ale) n w am CLCQ(0 0307 q l g ) (0 0307 "pig) (00307 "gig) nCQ(0 0307 <LW(00307 "gig) CLOQ (0 0307 "gig) UW (0 0307 `pig) aoQ(0 0307 "pig) uw aOQ(0 0307 "gig) (00307 "gig) aw < L o p (0 0307 "gig) (0 0307 "dg) <W (00307 ugip) noo (0 0307 "PIF) Hem PFOSEA "gig UW 4 r*? RSD Std I*r M M S D RPD NA NA NA N 4 Analytical Report: FACT-TOX-001 LRN-U2103 ETS-8-7 0 3M EnExcveli97ronmental Laboratory LITWCCI~O5 ( 2 ) TOX001-hver2l2-lCC XIS 5/3112WI 4 48PPMage 275 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance# 6329-212 Dale of Dab RcducliodAnalynl Sample Data 104 Weak Dlemy Canmogerucdy Sfudy with N-u Range (98 1%) N-Ethyl PerfluomarlanenulfonarmdoEthanol in Rats T-6316 (EIFOSE-OH) Rat Liver Filenam See Altachmenfi ETS-8-60 d ETS-8-10 R - S q w d Valva See Allachmeots Amlia 062498, Darey 070799 SlOpC Sce Allnshmnls MnssLp 3 3 Y-Inlmopt See Attachments 02/14/00.02116/00 RWWISAL 0221IW. 02/29lW, 03/0liW. 03,031W. 03104iW.071281W. 08ll51W MMSCWE54.5 02/281W,01101/W, 03/03100,03/06/W,07/31/W. 08/17/W LASiUMH WEEK 105 RAT LI R sample n PfOS CdC. cone. GGmmUuDp 3 Mid Dose 30 0 mgkg C90842M C90844M C90850M wg 156858 42289 46376 C90851M c90854M C90855M C90856M C90858M C908M1M C90864M C9086SM C90867M C90869M C90871M C90X81M C908X2M C90883M C90884M 120793 179256 73931 16276 131992 337224 122357 74361 102548 42WI 90688 42229 79628 38587 61036 C908XSM C90889M C90893M 102254 36130 58405 omup 3 Mid Dou C90900M C91252F C91153F 26321 158371 IW973 30 0 m#he C91254F C91256F C91264F C91278F C91279F 217110 161881 143127 231922 39747 C91286F C91292F C91294F 227867 48865 I17280 C91295F C91301F 22079 84658 .r.ru>=rer,,uomart*nu C91303F C91307F C91310F ate 166184 56102 126168 PFOSA = Perflw-laoe. PFOSAA = Perfluomfar EIFOSE = N-w Ranp I MSJ6 =C8F17SOZN(o( :oO) PFOSEA = P e r f m m l a n c sulfonyl ethyl-& Date EntemdIAnalyrf 03114/~,03/16'00,03117100, 08!151W, Dale VcritiedImlyd 03/09101 haj pwity EnlcmdNeriTrsd OY19101 LAC coo.rot..tiia of PFOS uglg or Y. Rec. 157 42 3 46 4 I21 179 73 9 163 132 337 I22 74 4 103 42 0 91 42 2 79 6 38 6 61 0 102 36 1 58 4 26 3 158 101 217 162 143 232 39 7 228 48 9 117 22 I 84 7 166 56 I 126 091031W LAC RSD PMSA CWIC~~f..fiO~ Std Ikv Cdr. Cooc. afPFOSA MSMSDRPD ns/e uyg or K r(re. 7349 7 35 36% 3 70 6514 651 6828 6.83 9569 9 57 10434 IO 4 8293 8.29 6178 6 18 12569 12.6 9434 9 43 4441 4 44 5466 5 47 5521 5 52 8631 8 63 6403 6 40 7493 7 49 5439 5 44 10849 10 8 8430 8 43 6567 6 57 78 1 6949 6 95 901 70 3 6625 6 62 6992 699 7206 7 21 9190 9 19 7676 7 68 6251 6 25 8954 8 95 4670 4 67 9554 9 55 6960 6% 1509 7 51 3266 3 27 5559 5.56 6664 666 54 0 5896 590 I 27 - 68 5 ~ 5242 5 24 RSD sld D e MSIMSD RPD 29 1 2 17 25 4 I 72 PFOSAA Cak.COW. ogls 5840 6967 10194 6845 365 I 3 1%89 10390 9279 11573 10854 4761 5093 12412 11794 6916 8486 5935 17W2 17198 I5569 x459 9694 5114 8009 8292 10693 5070 ' 12207 4129 17499 6906 9383 5143 4470 8525 7067 6657 Eaoeenmtion OrPposIuI uyg or K b. 5 84 697 10 2 6 85 36 5 I9 7 10.4 9 28 II 6 10 9 4 76 5 09 124 11 8 6 92 8 49 5 94 17.0 17 2 IS 6 8 46 9 69 5 II 8 01 8 29 IO 7 5 07 12.2 4 73 17 5 6 91 9 38 5 I4 4 47 8.52 7 07 665 Analytical Report: FACT-TOX-001 LRN-U2103 RSD std Ik" MSMSDRID 60.8 6 95 43 6 2 de EtFOSE C.lC corn. 95.3 166 39 3 I58 39 6 970 32.9 65 2 80 0 75 2 135 81 2 75 7 48 8 58 0 I55 48 8 73 3 218 47 7 79 9 ow 76 2 105 98 5 98 I 104 129 23 8 IO7 I51 137 9 54 59 4 122 88 4 33 3 Conrrnlr.Hoo ofElFOSE uygorY.Rre. 0 0953 <LOy (0 0614 "gig) ~LCQ(00614u~g) a IS8 aw)(00614 wk) 0 0970 UoQ(OO614ug/g) 0.0652 0 0800 0 0752 0 135 00812 ~LCQ0(O07O56714ug/g) Mean EIMSE "gk <LcQ(oM14"#e) 0 I55 <Oy(O0614u#g) 0 0733 'La) (00.2016814 udg) 0 0799 aw)(O0614ug/g) 0 0762 0 105 0 0985 0 0981 0 104 0 I29 < L o p (0 0614 "gig) 0 107 0 151 0 137 SLCQ (0 0614 "gig) 0 0594 0 122 00883 RSD sld DW MSMSD RPD 47 1 00415 26 9 0 olio ETS-8-7 0 3M EEnxcveli97ronmental Laboratory L w Week IO5 (2) TOXa01-liucr212-I00XIS 513112001 4 48 PPM age 276 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covancdl6329-212 Sample Data WEEK 105 RAT L :R s.mpir x M556 C.l.2. Cow. n#g C90842M inns7 Md h C908MM 6798 6 80 CM8SOM 8480 8 48 CW851M 11631 I1 6 C90854M 16151 162 CM8SSM 8603 863 C90856M 4869 4 87 CM8SXM 11618 I1 6 C9086oM 15449 15 4 cwn64M 11779 11 n C90865M 78x8 7 89 CM867M 6286 6 29 C90869M 7985 799 CM871M 1n3~ 10 4 CW881M 6427 6 43 cmnm ins00 10 s CM883M 5327 5 33 C90884M 14763 1476 CM885M 12272 I? 3 C90889M 802s n 03 7724 7 72 CM900M 5398 5 40 Gmvp 3 C9IzS2F 9729 9 73 Mid Dore C91253F 8473 I47 30 o mene C912S4F 1s64.1 I5 7 c91256F 10275 10 3 C912MF 6689 6 69 C91278F 10199 10 2 C9127YF 446s 4 47 C91286F 1w51 IO 1 C91292F 9421 9 42 C91294F 10286 10 3 C91Z9SF 2961 2% C913OIF 7474 7 47 C91303F l2SZ2 1:s C91307F 5325 5 32 C9I3IOF SW6 s ni PFOS ~ Pcrfluomoclvles mte PFOSA = Perfluomoctmnl bnarm& PFOSAA ~ Pcrfluomaclanesulfondos~tate EtFOSE = Narmw Rango N-Ethyl Perfluomaelanenulfonamidoethyl alcohol MS56 =C8F17SOW((H)H2CoO) PFOSEA = Pcrfwmactane sulfonyl ethylamidr ~~i~~~ t ~ r r d / ~ n s i0~31~14tiw. O ~ I I O Wn,3/17iw, nniis!w, 09103iw LAC Date VctiliediAnaly~ 03/09/01 hoj Punty EnnteredNenlied OL'lYIOl LAC OM) 000 0 182 3 27 OW 34 6 3 37 3 29 00 n 932 0214 000 OW 3 87 2 54 OW 3 22 ow 2 54 0.W 2 86 OW 38 8 I6 6 3 33 OM Analytical Report: FACT-TOX-001 LRN-U2103 NA NA NA 3M Environmental ETS-8-7 0 Excel 97 Laboratory Lvr Week 105 (2) TOXM)I-bver2l2-IW XIS S,3lrx!ill 4 4P8 PaM ge 277 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance# 6329-213 Analytical Report: FACT-TOX-001 LRN-U2103 EEK 1M RAT LI Cnup raw 1 S.mpkX 1"llhl W' I T.L.1r.f". dL*a PFOS s * C."=dh" Sung.* V"W - PFOS C.=. PmS C.*. cos. I Q.Ct. n ,0750 NA 9275 n I OM1 NA 9275 I0130 NA 09275 NA 867 NA 829 NA 250 IUh 321257 1,337. 313 98979 990 0 9881 NA 09275 09911 NA 0 9275 IOU8 NA 0 9275 I0776 NA 0 9275 I os2 NA 0 9275 ,0884 NA 0 9275 NA 8M 718021 718 NA 584 236132 236 NA 311 ,20112 I20 NA 70.6 26240 26 2 NA 7% 628332 628 NA 416 153299 I51 0 9935 NA IWW 10069 NA 09275 0 9872 NA 0 9275 I0290 NA 0 9275 I WII NA 0 9275 Hi&, Not Confirm4 561 NA 742 NA 464 NA I55 NA M3 565041 565 295155 295 186556 I87 60124 601 265167 265 ,0111 NA lwMl Hsgb. Not Co-fiimrd 407 201182 201 0 9%2 I1561 NA 0 9275 NA 0 9275 NA 857 689313 689 NA 642 ,4103, U5 0 993, NA 0 9275 NA 570 23w 230 C91132F C'II315F C9139f.P I 0 5989 0 9827 I I502 I1177 IO214 NA 09275 NA 09275 NA 0 9275 NA 0 9275 NA (I 9175 NA 635 NA 778 NA 542 NA 614 NA 841 509159 509 31737 I1 7 377842 178 220157 220 129837 I10 C91318F CPII4OF NA 0 9275 NA 09275 NA 851 3,IJSP 341 NA 718 579769 580 C9lJllF ,0145 NA 0 9275 NA 0 9275 NA 626 242581 243 NA 891 355109 355 CP1356F C91157F C913WF I0115 IO112 VA 0 9271 NA 0 9275 NA 0 9275 NA 21. NA .SI NA 522 41389 42 4 192%) 193 206612 207 C91361F NA 09275 NA 889 711319 712 C91362F ,0130 NA 0 9275 NA 220 868% 86 9 C91372F NA 0 9215 NA 639 257881 C91376F C91378P C91lBOF C91017M 0 9840 0 988, " NA 0 9275 NA 9275 NA 0 9275 NA 0 9275 NA 668 - NA 853 NA 797 NA 151 251932 14C458 324616 2014 C91054M I0156 NA 0 9275 NA 251 198 C91056M NA 0 9275 Lov,nolCoofimd 131 10494 C91060M I w62 NA 0 9275 C91065M 09918 NA 0 9275 NA 252 2w6 NA .I 2 16 5 CPlOiOM C91071M C11072M 0 9.1 0 9971 n986 NA U 9275 NA 0 9275 NA 0 927q NA 25. 205 NA JI 2 I, I YA 479 384 C11073M IWl6 NA 0 9275 NA I04 83 I C91076M I0102 NA 09275 Lgb. NolConfimd 613 502s C91451F I0161 NA 0 9275 195 I53% C914JIF IO133 NA 0 9275 572 ,525 C9145F 10117 NA 0 9l75 217 18803 C91459F C91462F ,0140 I W82 NA 0 9175 YA 0 9275 62 6 221 1W *9 4 17533 C91467F IO110 NA 0 9275 174 I 111 C91473F COI47PF 0 9879 I WII NA 0 9275 NA a 9275 171 IW I' s 13901 :c7 CPllSOF C9148SF C9I487F C9148PF 0 9882 NA 09812 NA - 10175 IO101 NA KA 0 9275 0 9115 n 9175 n 9275 183 I 251 50 - 19 I 143 10"WC MC8 149 10129 62 8 11169 Irntm. LAC ETS-8-70 3M EnL Cvd9i7ronmental Laboratory Page 278 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covance# 6329-212 - - PFOSA PFOSA - - COK DUIb. wr F.d.. NA 316 50 q'g .r x h. NA 316 50 NA 251 50 NA 367 50 I8 J NA 353 50 178 NA 84 9 50 406 NA 249 50 NA 358 50 116 NA 288 50 NA 2w 50 I5 I NA 198 50 A030I00080 NA I86 50 A030I0008I NA 206 50 NA 291 50 N* 176 50 - NA 407 50 NA - 384 50 16616 1 100 145 868 20 1 16 6 ~ NA 25. 50 128w 128 NA 324 50 16226 162 NA zm 50 1M27 10 6 NA 392 50 17062 A0301W094 I7 I NA 325 50 14551 A0301W095 1.6 NA 105 50 14949 A03OIwu% 11.9 NA 361 50 18058 A030low97 I8 I NA 319 50 16126 A0301W1w 16 I NA 116 50 15274 AO3OIWIOI 151 NA 338 10 16810 AU~QIWIO~ I6 B NA 125 50 6157 A0301W103 6 16 NA 336 50 16831 AO0301WIM 16 8 NA 364 50 17955 AO301WIO7 180 NA 446 50 22312 AOJOlMlOS 22 3 NA 220 50 10883 AOlOlWlW 109 NA 186 50 I9451 A0301WIIO 195 NA 3u 50 162% A0301Wlll 16 3 - - NA 380 50 18952 A00301Wll4 I t0 NA 303 50 NA 2 I4 I NA 2 50 I NA 43 0 I NA IS, I NA 272 I NA 171 I NA 2 52 I NA 2 67 I - NA 0 85 I NA I 76 I NA 166 I NA I31 I NA 1 66 L NA 0 380 I NA 2 51 I NA 2 71 I NA 2 30 I NA I62 I NA 0 870 I NA 22, I - - NA 0910 I NA . 2 91 I V.luc.8l ** h U n v A-5a-k Analytical Report: FACT-TOX-001 LRN-U2103 - PFOSAA - C.u. sir 459 392 5m 6% 475 37 3 333 819 403 157 358 226 377 5.8 95 5 l0Jl 622 ~ 314 564 758 585 408 402 ll8 537 524 613 149 413 577 631 608 SM 57s - 604 604 217 198 74 7 44 8 28 6 19 0 17 7 30 0 21 8 20 1 ~ ow OW ow OW OW ow 0.W "Y -"" I1 I 7 07 - OW I3 I - P m A h - mi.#*. F.U., 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 ~ 50 50 50 50 50 50 50 50 50 50 0 50 50 50 50 50 50 - 50 50 I I I I I I i I I ~ I I I I I I I I I - I I - P F O S U m*mm C.rnl.t",*" A Cmk.C o r 21358 A0301W066 dPFOSAA ugtgor K w. 21, 18476 A0301W067 I85 448.7 A0301W068 448 35222 A00301~9 352 23991% A0301wO72 23 9 1784 A0301WV73 178 15462 A0301wO74 I55 40328 A030100075 403 l8Sll A030100076 185 23W8 A030100079 230 17773 A030l00080 178 11445 AO3OlOM8l 114 I8340 A030100082 183 27358 A030100083 271 471 A030100086 472 51758 A030100087 518 26905 A030100088 269 ,579, A0030100089 158 28252 A0301wO90 281 38590 2548 AO301wO91 AO30IwO94 386 151 I8251 A0301Wu95 183 IS76 A0301KQ96 197 20954 AOlOlWu97 210 26299 AOJOlWlM 26.3 25334 AOJOlWlOl 253 33469 A030100102 33.5 17210 AUO33001I0W01IM03 20686 2107 72 28490 A030100107 28.5 31554 AO301W108 31 I 3W22 AO301W109 300 42332 X1., A030100110 AW3DIM111 423 271 - ,0114 30712 220 195 74 5 44 5 28 9 19 I ?3 I 30 I 21.8 20 I ow ow I OW OW A042800030 <LC6 0123 OW OM 0.M 000 112 7 21 - OW 12 9 MI.ll PFOSAA w, 23 6 RSD Sld Der MYMSD W D 56 5 133 26 3 7.08 1111 00614 NA NA 3M Environmental Laboratory Page 279 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covancd 6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 Sur..,.U V"iM NA NA NA NA NA N.4 N* YA NA NA NA NA NA NA N* NA NA NA NA N* NA NA NA NA NA N.4 NA NA hA NA NA NA NA NA NA NA NA NA NA NA NA NA NA ,.A NA NA NA NA NA NA NA NA NA NA NA NA NA NA - EtFOS -c.n. nr'. k69 293 813 464 331 ow 113 314 363 32 I 310 232 213 466 314 1020 594 ~ (39 377 274 4% 776 I79 916 416 746 182 88. 8.2 611 1106 211 518 774 - 1062 5.9 ow ow ow 000 "W ow om "W ow - ow ow ow ow ow ow uw ow ow ow ow - OW ow Y d V L tltl ** mbnu ElFOSE I EtFOSE I Flklrm I 110 A080100020 I 334 A080100021 I OW A080100024 I 291 AGBDL00025 I 309 AUII00026 0 3W I 31. A080100027 0331 I 323 AOSOI00028 0.323 I 308 A080100031 0.308 I 135 A080100032 0 235 I 207 AQt.0100033 0 207 I 466 A080100034 0 166 I 110 AD80100035 0310 I 1024 A080100038 I02 I 514 I 442 051. n 442 2 755 (1751 I 279 A080100042 I 432 AO801m015 2 1389 I 469 A0801W317 1 917 A080100018 I 408 A080100049 I 71I A08010N52 I 777 A0801W353 I 182 I 843 AOZOl00055 10 603. AOllJWlW I llu6 A080100059 I 211 I 522 A0801CW61 1 731 2 2117 A030400085 I 557 A0801M066 I OW A081400028 RSD S I 1 Drr MsiMSD RPD 57 8 0 229 131 I30 N* NA NA NA Sur"l.l VI- NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA HA NA NA NA NA NA NA NA NA NA N.4 Nk NA NA NA NA NA M% cor. Wh 623 633 619 688 641 177 5W 626 553 517 351 510 420 727 I88 8877 8666 457 653 193 710 552 553 614 636 542 839 228 516 307 889 480 789 717 475 656 270 318 OW Ow OW Ow OW ow ow "W M3% C.k.C.a. ol/l 28960 29855 30567 32781 32395 8k69 23216 30818 25101 27552 l7w6 25822 20389 36299 9313 US51 37172 22980 32766 9807 32164 24677 27071 30741 3ll5I 26187 41708 11248 16842 25014 lUhl 23712 39723 33977 23656 33352 273 3 I3 OW OW OM OW cc.3 OW Flklvmr AOIOIW066 A0101~7 AO301w068 A0301wo69 A030100072 A030100013 AO3OIWOIk A030100075 A030100076 A030100079 A030100080 A030100081 A030100082 A0301WO13 A030100086 A030I00087 A030100088 A030100089 A0301WOW A030100093 A03OIWW1 AOJOlWW5 A0301wo96 A030100097 AUIOIWIW AOJOlWlOl A0301W102 A03010(111)3 A0301W101 A0301WI07 . AO3OIOO108 A0301wIO* A0301M110 A0301Wlll AOIOIWII1 AOSOIWII5 A0811wO28 A081400029 A042800035 A042800016 A042800011 A04280WlS I *""%-:? A,;2800042328wUIZ AM2800026 OW A0428wo30 OW AO4280003l OW A0428W332 OW A042800033 A081400052 AO8I400055 29 9 30 6 32 8 32 1 8 47 23 2 30.8 25 1 17.6 I14 25 8 20.1 36 3 9.31 44 6 37 5 23 0 32 8 9.81 32 2 21 7 27.1 30 7 31 2 26 2 41 7 I1 2 26 I 25 0 1. I 23 7 39.7 RSD SM Drr MYMSD W D 34.7 9 11 ,I I 8 86 NA NA NA NA 3M Environmental Laboratory Page 280 3M Medical Department Study: T6316.1 AMDTU 092597.1 Covanc& 6329-212 C9IMIF c911.2F C91356F C91357F CP1160F c91361F C91362F C9l371P c91376P - S W n l - V I M NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA N* NA NA NA NA NA NA NA NA - NA N* NA NA NA NA NA NA NA Y4 NA NA NA NA NA NA NA NA NA NA NA NA -NA NA - QF0SE.A - C.*. Wk OW 21 7 34 I ow 2 39 50 1 37s 31 I 31 7 54.. ow 26 0 ow ow 111 21s 3 03 57 8 31.7 32 7 2 31 461 10 I 11 0 1.9 107 10 9 56 7 115 25 6 23 2 2 16 26 6 25 6 - 28 I 25 6 ow (I 00 ow 00 ow ow "W ow ow ~ ow ow ow ow OM ow ow ow OM ow ow - nw uw "" V d u r r s t l u.D *=sum,. - PFOSEA Csk. Cam. 0.W 20 5 33 7 OM 241 48.3 3 48 307 29 I 54 8 000 263 ow ow ,I5 236 - 2.62 58 I 32 7 33 3 2 03 4 12 10s 130 1, L 10 I IO 8 56 0 I15 25.2 23 2 2 82 26 8 2.3 - 28 6 26 0 ow OW am ow ow ow ow ow ow ow ~ ow ow ow ow ow ow ow "M ow ow - OW OM L I oc*r Ulit cvs YIrn'l 5'01. ".lyu Analytical Report: FACT-TOX-001 LRN-U2103 RSD S I 1 Drr W S D RPD I39 00566 57 6 om9 NA NA NA PA 3M Environmental Laboratory Page 281 3M Medical Department Study: T6316.1 AMDT# 092597.1 Covancdl6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 'LIVER bnpk # PFOS C.L. C.X. 111 990 718 236 120 161 628 I33 365 293 I87 CW953M 60 3 163 201 689 C91315F C91336F C91338F C91340F C91311F C91112F C9IJSLP C91357F C913WF C91361F C91362F 1 210157 329817 341389 579769 242581 355109 ,2189 191963 206612 712319 868% C91380F C9IO54M CPIO56M C9106OM CP1065M C91070M CilOiil 324616 l0.94 41 I I 38. C91453F C9I455F CP1b39F C91k62F C91467F 4525 I8803 I7533 137 C91b85F 1 1031858 325 203 n 198 105 201 0 0365 0 205 "a0411 $8. "0831 5 024960 I5 4 4 53 188 onw 175 0 137 13 9 L W i O 0245 ugtjl 0 I49 1033 00628 111 M"0 rms "8'8 313 11679 11920 12376 I8544 17812 I1539 11218 15070 14511 8680 17M2 14551 1.949 18058 16126 15274 ,6810 6157 16831 17955 22322 10883 19454 18952 I ,5385 2 li 246 429 I 83 2 71 I 267 ugh .r Jb b. 1.7 I. 9 I24 I83 178 406 11.5 176 I3 2 I5 I 9 82 941 ID0 11.5 8 68 20 I 166 128 I6 1 106 I7 I 146 I4 9 181 16 I 153 16 8 62 16 8 180 22 3 IO 9 1') 5 I6 3 I90 Rso suDe MYMSD RPD 31 3 4 23 23 I 3 61 NA NA NA NA 21358 44847 33222 23948 1784 134662 40321 18511 17773 11115 18340 27358 4725 51758 26905 I5794 18152 385W 23421 1825, 19676 2W51 16199 25331 11k69 ,7230 IMS6 28.W 3,551 3022 41332 27417 30114 30712 no IPS 74 5 14 5 28 9 I9 I OM OM OW OW U8 35 1 23 9 I78 1 I55 40 3 I I85 23 0 183 17 4 4 72 51 8 26 9 15.8 28.3 38 6 25 4 I83 19 7 21 0 26 3 25 3 33 5 172 20 7 28 5 31 6 30 0 12 3 27 1 30 I 30 7 a 2u1 no195 n 0745 0 0415 00289 00191 ""IW 00301 00218 "0201 'IW (0 0113 "ye <LW(00123 ads <Lcq(Oom"glg CLcQ(00123"dB cLcq(aol23y%ig <LW(OOl23u~lg <LW(00123 UYg .lCQ(O 0123 uwg CLOG (0 0123 udg r l W (0 0123 ug/g ( l W ( O O 1 2 3 uglg RSD Sld D.* MYMSD RPD 308 0 308 I 235 0235 207 0 207 I 466 0466 36 5 133 75s 179 131 1389 469 917 408 721 777 182 843 6034 0 755 0 279 0 432 I389 0.69 0.917 0 408 0721 0 777 0 182 0 843 6 03 I II 522 734 I 0522 073, 16 3 708 118 00614 NA N* 3M Environmental Laboratory Page 282 3M Medical Department Study: T6316.1 AMDT# 092591.1 CovanceU 6329-212 Analytical Report: FACT-TOX-001 LRN-U2103 R Swkl - MSY - Cak. Cow. ulh 28%0 29855 29 9 30567 30 6 32781 32.8 32395 32 4 8469 8.47 13116 21 2 30818 30 8 25WI 25 1 27552 27 6 17406 17, 25822 25 8 20389 20 6 36199 36.3 1313 931 us54 1.6 37472 37.3 22980 13.0 12766 318 9807 981 32164 32.2 14677 24.7 27071 27 I 30744 30 I 31154 26187 41708 11218 26812 25014 44467 23712 19723 011761 ,3977 CYI~F C9138OF C9IOOM - 23656 33352 2 73 3 I3 ow ow ow OM ow ow ow ow ow ow ow 0.w ow oca ow I36 ow 4 71 - I 1 9 293 7 MYMSD RPD 34 7 2.62 58 I 32 7 33 3 2 03 4 12 IO 5 13 0 26 I 103 10 8 I :"6 I 160 115 25 2 23 1 ow NA 0.w RSD Std Dr MYMSDRPD I39 0 OSM 57 6 00139 NA NA NA NA 3M Environmental Laboratory Page 283 3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 ~ Appendix F: Example Calculations Analytical Report: FACT-TOX-001 LRN-U2103 Analytical Report: FACT TOX-001 LRN-U2103 ~~ Formula Used for Sera Analyses in Study FACT TOX-001 AR (ng/mL) x DF x SC x PF x FV (mL) x 1.0 pg = Reported Concentration (pg/mL) EV (mL) 1000 ng Calculation Used for Group 2, Week 105, Animal ID C90783M, determination of PFOS 189.2 ng/mL x 10 x 0.9275 x 0.8640 x 1 mL x 1.0 pg = 1.52 pg/mL 1 mL 1OOOng AR-Analytical result from MassLynx summary DF-Dilution factor SC-PFOS salt correction constant (0.9275) PF-Purity correction factor FV-Final extract volume (1.O mL unless otherwise noted) EV-Volume of sera extracted Formula Used for Liver Analyses in Study FACT TOX-001 a AR (ng/g) x 3 curve (I) x SC x PF x DF x sample (') a curve is assumed to be: 1 g liver 5 mI, HZO 1.0 pg = Reported Concentration ,gg/g) 1000 ng Calculation Used for Group 2, Week 105, Animal ID C90783M, determination of PFOS 488.16 ng/g x 1 g / 5 mL, x 0.9275 x 0.8640 X 20 x 1.0 pg = 7.91 pg/g 0.9895 g/ 5 mL 1000 ng AR-Analytical result from MassLynx summary 3 curve-Density of the liver standard curve, assumed to be l g liver/ 5 ml water 3 sample-Density of the liver sample (g sample/ 5 mL H20) SC-PFOS salt correction constant (0.9275) PF-Purity correction factor DF-Dilution factor 3M Environmental Laboratory Page 284 3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Analytical Report: FACT TOX-001 LRN-U2103 Appendix G: Interim Certificates of Analysis 3M Environmental Laboratory Page 285 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 L m 3048 Research Drive Phone: (814) 231-8032 State College, PA 16801 Fax: (814) 231-1253 or (814) 231-1580 INTEMM CERTIFICATE OF ANALYSIS Revision 1(9/7/00) Centre Analytical Laboratories COA Reference #: 023-018B Test Name 3M Product: PFOS, Lot 171 Reference #: SD-009 Purity: 86.4% Specifications - Purity' Result 86.4% Appearance Identification NMR Metals (ICPIMS) 1. Calcium 2. Magnesium 3. Sodium 4. Potassium' 5. Nickel 6. Iron 7. Manganese Total % Impurity (NMR) Total % Impurity (LCiMS) Total % Impurity (GCNS) 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. Sulfate4 Organic Acids (IC) 1. TFA 2. PFPA 3. HFBA 4. NFPA Elemental Analysis': 1. Carbon 2. Hydrogen 3. Nitrogen 4. Sulfur 5. Fluorine White Crystalline Powder - Conforms Positive 1. 0.017 wt./wt.% 2. 0.007 wt./wt.% 3. 1.355 wt./wt.% 4. 6.552 wt./wt.% 5. 0.003wt./wt.% 6. 0.004 wt./wt.% 7. <0.001 wt./wt.% 1.oo wt./wt.% 10.60 wt.lwt.% None Detected 1. Theoretical Value = 17.8% 2. Theoretical Value = 0% 3. Theoretical Value = 0% 4. Theoretical Value = 5.95% 5. Theoretical Value = 60% 0.30 wt./wt.% None Detected Not Applicable3 1. c0.015 wt./wt.% 2. 0.27 wt./wt.% 3. C0.040 wt./wt.% 4. co.009 wt./wt.% 5. c0.006 wt./wt.% 6. C0.007 wt./wt.% 7. 8.82 wt./wt.% 1. co.1 wt./wt.% 2. co.1 wt./wt.% 3. co.1 wt./wt.% 4. C0.25 wt./wt.% 1. 12.08 wt./wt.% 2. 0.794 wt.lwt.% 3. 1.61 wt./wt.% 4. 10.1 wt./wt.% 5 . 50.4 wt./wt.% COA023-018B 3M Environmental Laboratory Page 1 of 3 Page 286 3M Medical Department -Study: T6316.1 Ilr. Analytical Report: FACT-TOX-001 LRN-U2103 lytical 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 Date of Last Analysis: OW31/00 Expiration Date: 08/31/01 Storage Conditions: Frozen 5-10C Re-assessment Date: 08/31/01 'Purity = 100% - (sum of metal impurities, 1.39% +LC/MS impurities, 10.60%+InorganicFluoride, 0.27%+NMR impurities, 1.OO%+ POAA, 0.30%) Total impurity from all tests = 13.56% Purity = 100% - 13.56% = 86.4% 2Potassiumis 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. 4Sulhrin the sample appears to be converted to so4 and hence detected using the inorganic anion method conditions. The anion result agrees well with the sulhr determination in the elemental analysis, lending confidence to this interpretation. Based on the results, the SO4 is not considered an impurity. 'TFA HFBA NFPA PFPA Trifluoroacetic acid Heptafluorobutyric acid Nonofluoropentanoic acid Pentafluoropropanoic acid '?heoretical value calculations based on the empirical formula, CgFF;7S03X+(MW=538) ' This work was conducted under EPA Good Laboratory Practice Standards (40 CFR 160). COA023-0 18B 3M Environmental Laboratory Page 2 Of 3 Page 287 3M Medical Department Study: T6316.1 I 3048 Research Drive Analytical Report: FACT-TOX-001 LRN-U2103 - -State Colleae. PA 16Rni INTERKM CERTIFICATE OF ANALYSIS Centre Analytical Laboratories COA Reference #: 023-018B LCMS Purity Profile: Impurity c4 c5 C6 c7 Total Note: The C4 and C6 values were calculated'usingthe C4 and C6 standard calibration curves, respectively. The C5 value was calculated using the average response factors fiom the C4 and C6 standard curves. Likewise, the C7 value was calculated using the average response factors fiom the C6 and C8 standard curves. ru#- Prepared By: Dayd S . Bell Date lytical Laboratories Date anager, Centre Analytical Laboratories COA023-0188 3M Environmental Laboratory Page 3 of3 Page 288 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 F4. 1 ,; t CEntrE Analytical Laboratories, Inc. 3048 Research Drive State College, PA 16801 Phone: (814) 231-8032 Fax: (814) 231-1253 or (814) 231-1580 Test Name Specifications Result 1. Carbon 2. Hydrogen 3. Nitrogen 4. Sulfur 5. Fluorine COA023-022-2 3M Environmental Laboratory 1. TheoreticalValue = 25.2% , 1. 25.04 wt.lwt.% 2. Theoretical Value = 1.75% 2. 1.69 wt.lwt.% 3. TheoreticalVaIue = 2.45% 3. 2.61 wt.lwt.% 4. TheoreticalValue = 5.60% 4. 8.88 wt./wt?h 5 . TheoreticalValue = 56.6% 5. 56.8 wt.fwt.% Page 1of3 Page 289 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 P 't CEntrE Analytical Laboratories, Inc. 3048 Research Drive State College, PA 16801 Phone: (814)231-8032 Fax: (814)231-1253or (814)231-1580 INTERLM CERTIFICATE OF ANAI,YSIS Centre Analytical Laboratories COA Reference #: 023-022-2 3M Product: EtF'OSE-OH Test Control Reference #: TCR-00017-52 Date of Last Analysis: 11/26/00 Expiration Date: 11/26/01 Storage Conditions: <-lo "C Re-assessment Date: 11/26/01 'P~ty= 100%- (totalNMR impurities, 1.26%+ GCMS impurities, 1.29 +POAA, 0.10%) - Total impurity from all tests = 2.65% Purity = 100% 2.65% = 97.4% 2~~~ Trifluoroacetic acid fl HFBA Heptafluorobutyric acid NFPA Nonafluoropentanoic acid PFPA . Pentafluoropropanoic acid 3Theoreticalvalue calculations based on the empirical formula, C ~ ~ H ~ O F ~ ~ N O ~ S (Mw=571) COAO23-022-2 3M Environmental Laboratory Page 2 of 3 Page 290 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 L m 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 #: 023-022-2 3M Product: EtFOSE-OH Test Control Reference #: TCR-00017-52 GCMS Purity Profile Peak # 1 2 Total Retention Time Wn) 13.934 17.307 Identity PFOSDEA c7 - % Impurity 0.36 0.93 1.29 This work was conducted under EPA Good Laboratory Practice Standards (40 CFR 160). Prepared By: Scientist Centre Analytical Laboratories COAO23-022-2 3M Environmental Laboratory ///6dd Date Page 3 o f 3 Page 291 3M Medical Department Study: T6316.1 I -.-. .. .. _ _ Analytical Report: FACT-TOX-001 LRN-U2103 CEntrE Analytical Laboratorks, Inc. P\ c 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#: 023-022-1 3M Product: EtFOSE-OH Test Control Reference fir: SD-013 Purity: 88.9% 1. Calcium 2. Magnesium 3. sodium 4. Potassium 5. Nickel 6. Iron r? 2. Fluoride 3. Bromide 4. Nitrate 2. Hydrogen 3. Nitrogen 4. Sulfu COA023-022-1 3M Environmental Laboratory 1. <0.001 wt.fwt.% 2. co.001 wt.fwt.% 3. <0.001 wt.lwt.% 4. 0.002 wt.lwt.% 5. <0.001 wt.lwt.% 1. Theoretical Value = 25.2' 2. Theoretical Value = 1.75% 3. Theoretical Value = 2.45% 4. Theoretical Value = 5.60% 1. <0.015 wt.lwt.% 2. C0.005wt.lwt.% 3. C0.040wt.lwt.% 4. <0.009 wt./wt.% 5 . dl.006wt.fwt.% 6 . ~0.007wt.fwt?h 1. co.1 wt.lwt.% 2. co.1 wt.fwt.% 3. <0.1 wt.lwt.% 1. 24.42 wt.fwt.% 2. 1.78wt.lwt.% 3. 2.72 wt.fwt.% Page I of3 Page 292 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 CEntrG Analytical Laboratorks, Inc. 3048 Research Drive Phone: (814)231-8032 State College, PA 16801 Fax: (814) 231-1253or (814)231-1580 7-3 RVTEMM CERTIFICATE OF ANALYSIS Centre Analytical Laboratories COA Reference # 023-022-1 3M Product: EtFOSE-OH Test Control Reference # SD-013 Date of Last Analysis: 11/26/00 Expiration Date: 11/26/01 Storage Conditions: <-lo "C Re-assessment Date: 11/26/01 'purity = 100% - (total metal impurities, 0.002% + total NMR impurities, 0.90% + GCMS impurities, 10.21 + POAA,0.03%) Total impurity fiom all tests = 11.14% Purity = 100% - 11.14%= 88.9% 2~~~ Trifluoroacetic acid r" HFBA Heptafluorobutyric acid NFPA Nonafluoropentanoic acid PITA Pentafluoropropanoic acid 3Theoreticalvalue calculationsbased on the empirical formula, C ~ ~ H ~ O F ~ ~ N O ~ S (MW=5 71) COA023-022-1 3M Environmental Laboratory Page 2 of 3 Page 293 3M Medical Department Study: T6316.1 - I L Analytical Report: FACT-TOX-001 LRN-U2103 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 #: 023-022-1 3M Product: EtFOSE-OH Test Control Reference #: SD-013 GCMS Purity Profile 11 16.379 c5 12 16.801 C6 - - 13 17.222 c7 Total 1.07 3.30 0.93 10.21 This work was conducted under EPA Good LaboktoryPractice Standards (40 CFR 160). Prepared By: Scientist ratories Centre Analytical Laboratories COAO23-022-1 3M Environmental Laboratory Date Page 3 Of3 Page 294 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 W ANALYTICAL REPORT STUDY TITLE PURITY DETERMtNATION OF SAMPLE LOTS OF PFOS Sample # TCR-00065-022 DATA REOUIREMENTS Test Article Characterization STUDY DIRECTOR "THISISAN EXACTCOW OF THE ORIGINAL DOCUMENT" Kevin Lloyd ANALYTICAL REPORT COMPLETION DATE October 25,2000 W; PERFORMING LABORATORIE/ TESTING FACILITIES Centre Analytical Laboratories, Inc. (Centre) 3048 Research Drive State College, PA 16801 Phone: 814-231-8032 STUDY SPONSOR 3M Environmental Technology and Safety Services Building 2-3E-09 PO Box 33331 St. Paul, MN 55133-3331 PROJECT IDENTIFICATION Centre Study Number: 023-045 Total Pages: 15 3M Environmental Laboratory Page 295 --3MMedicalDepartmentStudy: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Centre Study No.: 023-045 W GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT Centre Study Number 023-045, entitled "PurityDetermination of Sample Lots of PFOS, Sample # TCR-00065-022" conducted for 3M Environmental Laboratory, was performed in compliance with US EPA Good Laboratory Practice Standards (40 CFR Part 160) by Centre Analytical Laboratories,Inc. with the following exceptions: The automated data collection systems used in this study were not fully compliant with 21 CFR 58.130 (e). Kevin Lloyd Date Study Director Centre Analytical Laboratories,Inc. L/ Date Spohkor Representative 3M Environmental Technology and Safety Services :. . . > . .. , . . . . . SV- . .. , . Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 2 of 15 Page 296 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Centre Study No.: 023-045 w QUALITY ASSURANCE STATEMENT Centre Study Number 023-045, entitled "Purity Determination of Sample Lots of PFOS, Sample # TCR-00065-022"was reviewed by Centre Analytical Laboratories' Quality Assurance Unit. All reviewed phases were reviewed for conduct according to Centre Analytical Laboratories' Standard Operating Procedures, the Study Protocol, and ail applicable Good Laboratory Practice Standards. All findings were reported to the Study Director and to management. &% 1. Protocol Review Date Inspected 10119/00 Date Reported to Study Director and Centre Management 10/25/00 Date Reported to Sponsor Manaaement 10/25/00 2. Raw Data Review 10/19/00 10/25/00 10/25/OO V 3. Report Review 10/24/OO 10/25/00 10/25/00 Wiliiam spare Quality Assurance Officer /o z x o* Date "THIS IS AN EXACT COPY OF THE ORIGINAL DOCUMENT" W Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 3 of 15 Page 297 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Centre Study No.: 023-045 CERTIFICATION OF AUTHENTICITY This report, for Centre Study Number 023-045, is a true and complete representation of the raw data for the study. Submitted by: Centre AnaIytical Laboratories, Inc. 3048 Research Drive State College, PA 16801 (814) 231-8032 Study Director, Centre: Kevin Lloyd Date Study Director Centre Analytical Laboratories, Inc. Centre Analytical Laboratories, Inc. Facility Management: / Richard A. drazzi President Centre Analytical Laboratories,Inc. Date 'WE IS AN EXACT COW OF THE ORIGINAL DOCUMENT- V Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 4 of 15 Page 298 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Centre Study No.: 023-045 W STUDY IDENTIFICATION PURITY DETERMINATIONOF SAMPLELOTS OF PFOS Sample # TCR-00065-022 TYPE OF STUDY: Characterization TEST SYSTEM: Not Applicable TEST ARTICLE: PFOS,Test Control Reference # TCR-00065-022 SPONSOR: 3M EnvironmentalTechnology and Safety Services Building 2-3E-09 PO Box 33331 St. Paul, MN 55133-3331 W STUDY DIRECTOR. Kevin Lloyd CenterAnalytical Laboratories,Inc. Phone:(814) 231-8032 TESTING FACTLITlES: Centre Analytical Laboratories,Inc. (Centre) 3048 Rcsearch Drive State College, PA 16801 Phone: 814-231-8032 ANALYTICAL PHASE TIMETABLE: Study Initiation Date: Analytical Start Date: Analytical Termination Date: 09/28/00 10/05/00 10/24/00 "THISI S AN EXACT COPY OF THE ORIGINAL DOCUMENT" BY KT6 DATE ,&&e Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 5 of 15 Page 299 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Centre Study No.: 023-045 PROJECT PERSONNEL The Study Director for this project was Kevin Lloyd at Centre Analytical Laboratories, Inc. The following personnel from Centre Analytical Laboratories, Inc., were associated with various phases of the study: Name Gerry Shero David S. Bell Emily R. Stauffer Mark Ammerman & T Scientist Scientist Scientist Sample Custodian "THIS IS AN EXACT COPY OF THE ORIGINAL DOCUMENT" Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 6 of 15 Page 300 3M Medical Department Study: T6316.1 . . Analytical Report: FACT-TOX-001 Centre S t u d y No . . LRN-U2103 023-045 TABLE OF CONTENTS F&g TITLE PAGE ...................................................................................................................... 1 GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT ............................. 2 QUAL.ITY ASSURANCE STATEMENT.......................................................................... 3 CERTIFICATIONOF AUTHENTIClTY........................................................................... 4 STUDY IDE"ICATI0N ............................................................................................... 5 PROJECT PERSONNEL.................................................................................................... 6 TABLE OF CONTENTS .................................................................................................... 7 1.0 SUMMARY............................................................................................................... 8 2.0 INTRODUCTION......................................................................................................... 8 3.0 TEST SYSTEM ............................................................................................................ 8 4.0 TEST ARTlCLE............................................................................................................ 8 5.0 EXPERIMENTALPROCEDURES ............................................................................. 9 6.0 RESULTS AND DISCUSSION.................................................................................. 10 8.0 CIRCUMSTANCES THAT MAY HAVE AFFECTED THEDATA.................... 10 9.0 RETENTION OFDATA AND S A M P L E S ............................................................ 10 Table I. Results From LCNS forTCR-00065-022........................................... 11 w . Figure 1.PFOS Calibration Standard (Cl00500-4)at 269pgL .............................1.2 Figure 2.PFOS Calibration Standard (ClOO500-2) at 539pgL .............................. 13 Figure 3. PFOS (TCR-00065-022) at 25Opg/L. ESI Negative Ion Mode ...................... 14 Appendix A: Study Protocol OOP-023-045:PURITY DETERMINATION OF SAME'E LOTS OF PFOS. Sample # TCR.00065.022. Including Amendment 1.........15 '7HIS IS AN EXACT COPYOF THE ORIGINAL DOCUMENT Centre Analytical Laboratories. Inc. 3M Environmental Laboratory Page 7 of 15 Page 301 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Centre Study No.:023-045 purity). 1.0 SUMMARY m sampleffTCR was determinedfrom the E M S pure as compared with the PFOS control (TCR-00017-046,97.9% 2.0 INTRODUCTION This report details the results of the analysis of PFOS,Test Control Reference# TCR- 00065-022. The test was analysis by liquid chromatography- mass spectrometry (LCMS). The LCMS was conducted at Centre. The study was initiated on September28,2000 when the Study Director signed protocol number OOP-023-045.The analytical start date was October 5,2000, and the experimental termination date was October 9,2000. 3.0 TEST SYSTEM W There is no test system associated with this characterization study, therefore the GLP requirement for test system description,justification, and identificationdo not apply. The route of administration, levels and frequency of administration also do not apply to characterization studies. 4.0 TEST ARTICLE "THISIS AN EXACTCOPY OF THE ORIGINAL DOCUMENT" BY . s r f D A . L & i ? The test article was PFOS,Test Control Reference# TCR-00065-022. 3M Environmental Laboratory supplied the test article and it was logged at Centre Analytical Laboratories, Inc. as follows: Compound PFOS TCR-00065-022 Lot or NB Number t Centre Control No. 00-023-068 Puritv TBD Date Received 10/05/OO The sample received was a white, crystalline solid. It was received under frozen conditions and was stored in a temperature-monitored freezer kept at <-1O"C. W Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 8 of 15 Page 302 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Centre Study No.: 023-045 The analyticalkontrol standard PFOS was: Control Article Lot Number PFOS TCR-00017-046 Centre Control No. 00-023-042 Punty 97.9% - Expiration Date 8/3 112001 The chemical and physical data for PFOS is as follows. Common Name: Molecular weight: CAS Number: Structure: PFOS 499 (c~F17so3-) 2795-39-3 II0 II cfi17s 0-0'K' 0 ?.ate: the neutral mc,2cule and standard form that L,e PFOS \&an) is derived from is potassium perfluorooctanesulfonate ( C ~ F I ~ S O ~mKo)le, cular weight 538. 5.0 EXPERIMENTAL PROCEDURES LCMS Spectral Analysis PFOS sample TCR-00065-022was analyzed for purity using liquid chromatography/mass spectrometry(LCMS). The sample was analyzed accordingto the following procedure. The samplewas prepared at about 250 &mL in methanol and analyzed by LCMS using a Hewlett-Packard 1100 HPLC system interfaced with a Hewlett-Packard 1100 mass selective detector. The mass spectrometer was run in negative ionization mode using electrospray ionization @SI) using Selective Ionization Monitoring for ion m/z 499. The liquid chromatography system was operated in reversed-phase mode using a C 18 silica-based column. Each resulting chromatogram was calculated against the analytical control article (TCR- 00017-046) calibration curve. "THIS IS AN EXACT COPY OF THE ORIGINAL DOCUMENT" V Centre Analytical Laboratories,Inc. 3M Environmental Laboratory Page 9 of 15 Page 303 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Centre Study No.: 023-045 6.0 RESULTS AND DISCUSSION LCMS Spectral Analysis Responses were observed for the PFOS sample in negative ion mode using an ESI interface. Quantitation was performed using SIM mode, mlz 499, and PFOS was detected at 7.8 minutes. TH-PFOSwas used as the internal standard,m/z 427, and was detected at 7.5 minutes. The total percent purity, calculated against the analytical control article (TCR-00017-046c)alibration curve was determined to be 88.0% from the average of three replicate analyses. 8.0 CIRCUMSTANCES THAT MAY HAVE AFFECTED THE DATA Electronic records are not fully compliant with 21 CFR 11,"Electronic records; Electronic Signature." However, approved SOPSwere in place and all instrumentation used in this study was fully calibrated and operational. All original raw data were printed as hard copies and fully audited by quality assurance. Verified exact copies and the electronic data will be stored in the archives at Centre Analytical Laboratories. Original raw data will be returned to the Sponsor. 9.0 RETENTION OF DATA AND SAMPLES When the final report is final, all original paper data generated by Centm 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 we11 as a signed copy of the final analytical report and all original facility-specificraw data, will be retained in the Centre Analytical Laboratories,Inc. archives for the period of time specifiedin 40 CFR Part 160. Retained samples of reference substances are archived by the sponsor. `THIS IS AN EXACT COPY OF THE ORIGINAL DOCUMENT" Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 10 of 15 Page 304 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Centre Study No.:023-045 Table I. Results From L C M Of TCR-00065-022,ESI Negative mode c 3M ID TCR-00065-022 TCR-00065-022 TCR-00065-022 Retention Time (min) 7.79 7.80 7.80 Mass ~ 499 499 499 Standard Standard Standard Standard 7.69 499 7.73 499 7.77 499 7.79 499 Value Pn/L 250 250 250 11 11 54 54 Determined Pa 219 220 220 % Recovery 88.0 88.0 88.0 loo* * 17* 1 * 60 110 50 93 Standards Average = Std Dev = 7 * First two injections during instrument warm up, not reported. CCV = standard in run used to check calibration (C100500-4 at 269pgL) "THISIS AN EXACTCow Of THE ORIGINAL DOCUMENT" Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 11 of 15 Page 305 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Centre Study No.: 023-045 ... ...' , ' ;... ., . .. ...'.^. '. .: h .. . , ' W Instrument 1 Mon, 23. Oct. 2000 00:12:06 pm Pa94 1 af 1 ';.. ---- 6;. ro/bs]Ld . 'THIS IS AN EXACT COPY OF THE ORIGINAL DOCUMENT" I . _.__ .e....-.*.A. -.,..-. >. . , BY " '*_ Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 12 of 15 Page 306 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Centre Study No.: 023-045 Figure 2. PFOS Calibration Standard (ClOOSOO-3) at 539pg/L c Sample Name:C100500-3 Sample Info: 1 Data f i l e :C:\HPC?IEM\1\DATA\100600\10060015.D ==l~lw=9~=lw9ll~lllLw~9D~.~~~~9~9w~~~9w~~~~~~w~w9wD~-9~w~w Injection Date P * A C o~perator Instrument Dilution : 10/6/2000 : OS GI ~ C : Instrument 1 :1 1:13:21 PM U Seq Line : Vial No. : Inj. No. : Inj. Vol. : 13 1s 1 5 P1 Acq. Method t PF0S.M Analyuie Method Last Changed I :\HPcHEM\1\METHODS\lOO6OO.M !0/9/2000 01:45:43 pm (modified after loading) Analysio or PFOS . I 1 Instrument 1 Mon, 9. Oct. 2000 01:46:00 pm V Centre Analytical Laboratories,Inc. 3M Environmental Laboratory Page 1 of 1 Page 13 of 15 Page 307 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Centre Study No.: 023-045 Figure 3. PFOS (TCR-00065-022)at 250pg/L, ESI Negative Ion Mode I Sample Name :L29092-1 Sample Ino:C100500-8 Data file :C:\HPCHEM\1\DATA\lOO600\10060019.D E=rlL-ll~r=ri~ri~irillllll~1llllllllllll9111111~~1~-=1111~~1~1~~g~~~~~g=~ Injection Date r A c q Operator Instrument Dilution : 10/6/2000 : GS 6% w&' : Instrument 1 :1 2:20:49 PM Seq Line : Vial No. : Inj. No. : Inj. Vol. : 17 21 1 5 Pl Acq. Method ; PF0S.M Analysin Method : C:\HPCHEM\l\METHODS\lOO600.M L a s t Changed : '10/23/2000 11:44:26 am Analysis for PFOS .st-- .. . m.BIM.mm:var ;( ! j j i i Instrument 1 Mon, 23. O c t . 2000 00:10:19 pm Page 1 of 1 "THIS ISAN EXACT COPY OF I THE ORIGINAL DOCUMENT" Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 14 of 15 Page 308 3M Medical Department Study: T6316.1 Analytical Report: FACT-TOX-001 LRN-U2103 Centre Study No.: 023-045 APPENDIX A Study Protocol Purity Determination of Sample Lots of U' PFOS Including Amendment 1 `?HIS IS AN EXACT COPY OF THE ORIGINAL DOCUMENT" V Centre Analytical Laboratories, Inc. 3M Environmental Laboratory Page 15 of 15 Page 309 3M Medical Department Study: T6316.1 3M Medical Department Study: T-6316.1 Appendix H: Report Signature Page Analytical Report: FACT-TOX-001 LRN-U2103 Analytical Report: FACT TOX-001 LRN-U2103 John L. Butenhoff, Ph.D., Study Director Date .Marvin T. Case, D.V.M., Ph.D., Sponsor Representative %w. *I Date 6</31 10 I Kristen J. Hansen, Ph.D., Principal Analytical lnvestigator Date William K. Reagen, Ph.D., Laboratory Manager Date 3M Environmental Laboratory Page 310