Document KzKzGxB2ODKw7Gr8104R23KKN

Northwest Bioanalytical Study No. NWBS00-145 Report No. NWBR01-155 Quantitative Determination of PFOS and Related Compounds in Human Serum by LC/MS/MS Sample Analysis Report for Protocol EPI-0016 Northwest Bioanalytical (NWB) A Division of NWT Ine. 1121 East 3900 South Salt Lake City, UT 84124 PREPARED FOR: 3M Company Corporate Occupational Medicine, Medical Department 220-3W-05 St. Paul, MN 55144 AUTHOR: Connie O. Sakashita, B.S., NWB Project Manager BY: _ Technical Director DATE: / - /<?- 2- DATE: ! /O ~0 Page 1 Northwest Bioanalytical Study No. NWBS00-145 Report No. NWBRO1-155 QUALITY ASSURANCE STATEMENT LABORATORY: Northwest Bioanalytical (NWB) A Division of NWT Inc. 1121 East 3900 South Salt Lake City, UT 84124 SPONSOR: 3M Company Corporate Occupational Medicine, Medical Department 220-3W-05 St. Paul, MN 55144 COMPOUND(S): PFOS and Related Compounds NWB STUDY NUMBER: NWBS00-145 SPONSOR STUDY NUMBER: EPI-0016 NWB STUDY TITLE: Quantitative Determination of PFOS and Related Compounds in Human Serum by LC/MS/MS The clinical study described in this report is not included within the definition of a GLP regulated nonclinical study. However, Northwest Bioanalytical conducts all studies within the ) guidelines of the U.S. FDA Good Laboratory Practice Regulations for Nonclinical Laboratory Studies (Title 21 CFR Part 58), the OECD Principles of Good Laboratory Practice and the Japanese MHW Good Laboratory Practice Standard Ordinance for Nonclinical Laboratory Studies on the Safety of Drugs (Ordinance No. 21, PAB Notification No. 424.) The following inspections were performed by the NWB QAU per SOP. Inspection and Reporting Statement. Date of Inspection Phase of Study Date Report Sent to Date Report Sent to NWB Project Manaeer NWB Management* 22-23 Oct 2001 22-23 Oct 2001 24 Oct 2001 15-16 Nov 2001 07-09 Jan 2002 10 Jan 2002 Standards Preparation QC Preparation Analytical Plan Sample Analysis Report Draft/Raw Data Final Report 23 Oct 2001 23 Oct 2001 24 Oct 2001 16 Nov 2001 09 Jan 2002 10 Jan 2002 31 Oct 2001 31 Oct 2001 31 Oct 2001 30 Nov 2001 31 Jan 2002 31 Jan 2002 *Reports to NWB Management are issued monthly. As can reasonably be established, the methods and procedures described and the results incorporated into this final report accurately reflect the raw data. t i NWB Quality Assurance Compliance Auditor Page 2 Northwest Bioanalytical Study No. NWBSOO-145 Report No. NWBR01-155 COMPLIANCE STATEMENT The clinical study described in this report is not included within the definition o f a GLP regulated nonclinical study. However, to the best o f our knowledge, this project was conducted in accordance with the guidelines o f the U.S. FDA Good Laboratory Practice Regulations for Nonclinical Laboratory Studies (Title 21 CFR Part 58) and according to the methods and procedures described within this report. In addition, the study followed the guidelines o f the OECD Principles o f Good Laboratory Practice and the Japanese MHW Good Laboratory Practice Standard Ordinance for Nonclinical Laboratory Studies on the Safety o f Drugs (Ordinance No. 21, PAB Notification No. 424.) Any known incidents that may have affected the quality or integrity o f the project or reported data is included in this report. This report represents an accurate record o f the raw data. /^r>c/o<u 0 Connie O. Sakashita, B.S. NWB Project Manager Date Page 3 Northwest Bioanaiytical TABLE OF CONTENTS Study No. NWBSOO-145 Report No. N W BR 0M 55 SIGNATURE PAGE................................................................................................................................... 1 QUALITY ASSURANCE STA TEM EN T............................................................................................. 2 COMPLIANCE STATEM ENT................................................................................................................ 3 TABLE OF CONTENTS...........................................................................................................................4 LIST OF TA BLES...................................................................................................................................... 4 LIST OF FIGURES.................................................................................................................................... 5 1. INTRODUCTION...............................................................................................................................8 2. METHODOLOGY.............................................................. 9 3. SAMPLE A N A LY SIS.....................................................................................................................12 4. RESULTS AND D ISCU SSIO N .................................................................................................... 13 5. REFEREN CES................................................................................................................................. 14 6. DATA RETEN TIO N ....................................................................................................................... 15 LIST OF TABLES Table 1. Calibration Curve Summary for PFO S..................................................................................16 Table 2. Calibration Curve Summary for P F O A ............................................................. 17 Table 3. Calibration Curve Summary for PFH S.................................................................................. 18 Table 4. Calibration Curve Summary for PFO SA A ........................................................................... 19 Table 5. Calibration Curve Summary for PFO SA .............................................................................. 20 Table 6. Calibration Curve Summary for M 556................................................................................. 21 Table 7. Calibration Curve Summary for M 570................................................................................. 22 Table 8. Back-Calculated Concentrations o f Calibration Standards for P F O S .............................23 Page 4 Northwest Bioanalytical Study No. NWBS00-145 Report No. NWBROl-155 Table 9. Back-Calculated Concentrations o f Calibration Standards for PFOA..............................24 Table 10. Back-Calculated Concentrations o f Calibration Standards for PF H S ............................25 Table 11. Back-Calculated Concentrations o f Calibration Standards for PFOSAA...................... 26 Table 12. Back-Calculated Concentrations o f Calibration Standards for PFOSA..........................27 Table 13. Back-Calculated Concentrations o f Calibration Standards for M 5 5 6 ............................28 Table 14. Back-Calculated Concentrations o f Calibration Standards for M 5 7 0 ............................29 Table 15. Analytical QC Summary for PFOS...................................................................................... 30 Table 16. Analytical QC Summary for PFO A ..................................................................................... 31 Table 17. Analytical QC Summary for PFHS...................................................................................... 32 Table 18. Analytical QC Summary for PFO SA A ............................................................................... 33 Table 19. Analytical QC Summary for PFOSA.................... 34 Table 20. Analytical QC Summary for M 556...................................................................................... 35 ) Table 21. Analytical QC Summary for M 570.......... ........................................................................... 36 Table 22. Serum Study Sample Concentrations.................................................................................. 37 LIST OF FIGURES Figure 1. Representative Calibration Curve for P F O S .......................................................................45 Figure 2. Representative Calibration Curve for PFO A ..................... 45 Figure 3. Representative Calibration Curve for P F H S .......................................................................46 Figure 4. Representative Calibration Curve for PFOSAA................................................................. 46 Figure 5. Representative Calibration Curve for PFOSA.................................................................... 47 Figure 6. Representative Calibration Curve for M556 .......................................................................47 Figure 7. Representative Calibration Curve for M 5 7 0 .......................................................................48 Figure 8. Human Plasma Blank for PFO S........................................................................................... 49 Page 5 Northwest Bioanalytical Study No. NWBSOO-145 Report No. NWBRO1-155 Figure 9. Human Plasma Blank for P F O A ..........................................................................................50 Figure 10. Human Plasma Blank for PFH S......................................................................................... 51 Figure 11. Human Plasma Blank for PFO SA A ...................................................................................52 Figure 12. Human Plasma Blank for PF O SA ...................................................................................... 53 Figure 13. Human Plasma Blank for M 556......................................................................................... 54 Figure 14. Human Plasma Blank for M 5 7 0 ......................................................................................... 55 Figure 15. Human Plasma Blank with Internal Standard (QCO) for PFO S.....................................56 Figure 16. Human Plasma Blank with Internal Standard (QCO) for PFOA.................................... 57 Figure 17. Human Plasma Blank with Internal Standard (QCO) for PFH S.................................... 58 Figure 18. Human Plasma Blank with Internal Standard (QCO) for PFOSAA............................... 59 Figure 19. Human Plasma Blank with Internal Standard (QCO) for PFO SA ................................. 60 Figure 20. Human Plasma Blank with Internal Standard (QCO) for M 556.................................... 61 Figure 21. Human Plasma Blank with Internal Standard (QCO) for M 570.....................................62 Figure 22. Low Standard for PFOS...................................................................................................... 63 Figure 23. Low Standard for PFO A ........................................................... ..........................................64 Figure 24. Low Standard for PFHS....................................................................................................... 65 Figure 25. Low Standard for PFOSAA................................................................................................. 66 Figure 26. Low Standard for PFOSA.................................................................................................... 67 Figure 27. Low Standard for M556....................................................................................................... 68 Figure 28. Low Standard for M570................................................................................................ 69 Figure 29. High Standard for PFO S...................................................................................................... 70 Figure 30. High Standard for PF O A ..................................................................................................... 71 Figure 31. High Standard for PFH S...................................................................................................... 72 Figure 32. High Standard for PFOSAA................................................................................................73 Page 6 Northwest Bioanalytical Study No. NWBS00-145 Report No. NWBRO1-155 Figure 33. High Standard for PFOSA................................................................................................. 74 Figure 34. High Standard for M 556.................................................................................................... 75 Figure 35. High Standard for M570.................................................................................................... 76 Page 7 Northwest Bioanalytical Study No. NWBSOO-145 Report No. NWBRO1-155 Quantitative Determination of PFOS and Related Compounds in Human Serum by LC/MS/MS Sample Analysis Report for Protocol EPI-0016 1. IN T R O D U C TIO N This report summarizes the analytical results from the quantitation o f PFOS, PFOA, PFHS, PFOSAA, PFOSA, M556, and M570 in human serum samples for 3M Company (3M) in support o f Protocol EPI-0016 [5.1]. The LC/MS/MS method for the analytes has a targeted calibration curve range o f 1.00 to 500 ppb (except for M556 which has a LLOQ o f 2.50 ppb). The actual calibration curve range for each analyte varies based upon the persistent levels o f the analyte in the matrix. For this study, the assay exhibited the following LLOQ and ULOQ values: A nalyte PFOS PFOA PFHS PFOSAA PFOSA M556 M570 LLOQ 3.37 ppb 1.44 ppb 1.36 ppb 1.50 ppb 1.00 ppb 2.50 ppb 1.00 ppb ULOQ 413 ppb 481 ppb 522 ppb 501 ppb 500 ppb 500 ppb 500 ppb The sponsor was 3M Company (Corporate Occupational Medicine, Medical Department, 220-3W-05, St. Paul, MN 55144), and the Study Director was Jeffrey H. Mandel, M.D., M.P.H. The Study Contact at 3M was Jean M. Burris. James K. Lundberg was the technical contact at 3M. The following is a list o f NWB supervisory personnel involved in the completion o f this work: Connie O. Sakashita, B.S. (NWB Project Manager); Patrick Bennett, M.S., M.B.A. (NWB Laboratory Director); Rodger L. Foltz, Ph.D. (NWB Technical Director). Laboratory personnel involved in the completion o f this Page 8 Northwest Bioanalytical Study No. NWBSOO-145 Report No. NWBR01-155 study included Suzanne Newman, B.S. (NWB Research Scientist); Laura Struhs, B.S. (NWB Scientist) and Anna Akrami, B.S. (NWB Associate Research Scientist). NWB SOPs and guidelines were used in the conduct o f this study and were available to study personnel in electronic and/or hard copy formats. Date Study Initiated: 23-Qct-2001 Date Analyses Completed: 16-Nov-2001 The clinical study described in this report is not included within the definition o f a GLP regulated nonclinical study. However, Northwest Bioanalytical conducts all studies within the guidelines o f the U.S. FDA Good Laboratory Practice Regulations for Nonclinical Laboratory Studies (Title 21 CFR Part 58), the OECD Principles o f Good Laboratory Practice and the Japanese MHW Good Laboratory Practice Standard Ordinance for Nonclinical Laboratory Studies on the Safety o f Drugs (Ordinance No. 21, PAB Notification No. 424). Any changes to or deviations from the original protocol (Analytical Plan) were documented through approved protocol amendments or deviation memos and are retained within the raw data. 2. M ETHODOLOGY ' The assay used for this study is reported in Northwest Bioanalytical reports NWBR00108 [5.2] and NWBR00-122 [5.3]. Chinese human plasma was used for the calibration curves and quality control sam ples. Serum sam ple results obtained using plasma curves are discussed in NWBR00-122. Samples for this study were received at NWB on the following date: Receipt Date 23-Oct-2001 Number o f Samples Received 262 Storage Condition (except during analysis) -20 C ) Page 9 Northwest Bioanalytical Reference Material Study No. NWBS00-145 Report No. NWBROl-155 Analyte PFOS PFOA PFHS PFOSAA PFOSA M556 M570 THPFOS Lot Number Purity Expiration Date Source 193 100% 31-D ec-2010 3M 245 100% 31-D ec-2010 3M SE-036 100% 01-Jan-2010 3M 617 53.8% 31-D ec-2010 3M 214 100% 31-Dec-2010 3M NB113047-8D 99.89% 31-Dec-2010 3M 118506-26 99.75% 31-Dec-2010 3M 59909 90% 31-D ec-2010 3M Stored dry Storage Conditions Room temperature* Room temperature* -20 C Room temperature Room temperature Room temperature Room temperature Room temperature The reference material purity for PFOS, PFOSA, PFHS, and PFOA was not available prior to the conduct o f this study. Therefore, the reference material purity was assumed to be 100%. 3M contracted with Centre Analytical Laboratories, Inc., in State College, Pennsylvania to determine the absolute concentration o f PFOS, PFOA, and PFHS in NWB stock solutions used to prepare the analytical standards and controls used for this studies. All arrangements for purity determinations and the transfer o f NWB solutions to Centre Analytical Laboratories, Inc., w ere performed by 3M . B ased on Centre Analytical Laboratories' results, the concentrations o f plasma calibrator and quality control samples were corrected using the following factors: Analyte PFOS PFOA PFHS Correction Factor 0.821 0.961 1.043 Eight or more calibration standards were prepared on the day of each run by adding 100 pL o f blank Chinese human plasma and 400 pL o f 50 mM ammonium acetate in Page 10 Northwest Bioanalytical Study No. NWBS00-I45 Report No. NWBROl-155 water (unadjusted pH ~6.9) to 13 x 100 mm polypropylene tubes. After a brief vortex mixing, 10.0 pL o f the appropriate spiking solution was added. The final calibration standard concentrations in human plasma for each analyte are listed below: Analyte PFOS PFOA PFHS PFOSAA PFOSA M556 M570 Calibration Standard Concentrations* 3.37, 4.60, 10.8, 23.1,43.6, 84.6,208, 331 and413ppb 1.44,2.88, 10.1,24.5,48.5, 97.1,241, 385 and 481 ppb 1.36, 2.92, 10.7, 26.4, 52.5, 104, 261, 417 and 522 ppb 1.50, 3.00, 10.5, 25.5, 50.5, 101, 251, 401 and 501 ppb 1.00. 2.50, 10.0, 25.0, 50.0, 100,250,400 and 500 ppb 2.50, 10.0, 25.0, 50.0, 100, 250, 400 and 500 ppb 1.00, 2.50, 10.0, 25.0, 50.0, 100, 250, 400 and 500 ppb * The target calibration curve range is 1.00 ppb to 500 ppb (except M556 which has a LLOQ o f 2.50 ppb). Each analyte has a different final curve range based upon the persistent levels o f the analyte in the human plasma used. In addition, blank human plasma samples, both with and without internal standard (designated as QCOs and Blanks, respectively), were assayed in each analytical run. Analytical QCs were prepared in Chinese human plasma on November 5, 2001 and stored in a -20 C freezer. For each run, analytical QC levels were assayed in duplicate. Analyte PFOS PFOA PFHS PFOSAA PFOSA M556 M570 Low 5.83 4.32 4.48 4.50 4.00 4.00 4.00 QC Concentrations (ppb)* M edium 126 144 156 151 . 150 150 150 High 331 384 417 401 400 400 400 * The target QC concentrations are 4.00 ppb, 150 ppb and 400 ppb. Each analyte has different QC concentrations based upon the persistent levels o f the analyte in the human plasma used. The internal standard (THPFOS) was added to all serum samples (except Blanks) for a final concentration o f approximately 200 ppb. Page 11 Northwest Bioanalytical Study No. NWBS00-145 Report No. NWBR01-155 The analytical method consisted o f a liquiddiquid extraction procedure followed by evaporation and reconstitution o f the extract residue with 20 mM ammonium acetate in water: 20 mM ammonium acetate in methanol (30:70 v/v). The samples were analyzed by liquid chromatography/tandem mass spectrometry using an API 3000. The instrument was operated in the multiple reaction monitoring (MRM) mode under optimized conditions for PFOS, PFOA, PFHS, PFOSAA, PFOSA, M556 and M570 detection o f the negative ions formed by turboionspray ionization. 3. SAMPLE ANALYSIS PFOS, PFOA, PFHS, PFOSAA, PFOSA, M556, M570, and THPFOS chromatographic peaks were integrated using the MacQuan software (version 1.6) with a smooth factor o f one. Quantitation was based upon quadratic regression analysis o f weighted (1/x ) calibration curves using area ratio vs. concentration utilizing Watson DMLIMS software (version 6.1.1.04). The samples were injected in a systematic order. ) 3.1. Acceptance Criteria For an analytical run to be accepted, it must have met the acceptance criteria listed below which are consistent with regulatory and industry recommendations. Calibration Curve Each run will include in duplicate calibration standards at six or more concentrations covering the lower to upper limit o f quantitation. For all analytes except PFOSA and PFOSAA, at least three-fourths o f the calibration standard's back-calculated concentrations must be within 15% ( 20% for LLOQ) o f their individual target concentrations. For PFOSA and PFOSAA, at least three-fourths o f the calibration standard's back-calculated concentrations must be within 20% ( 25% for LLOQ) of their individual target concentrations. A calibration standard will be considered an outlier if it is greater than two times the acceptance criteria for that standard. Page 12 Northwest ioanalytical Lower Limit o f Quantitation Study No. NWBS00-145 Report No. NWBROl-155 The back-calculated concentrations o f at least one o f the duplicate lowest points in the calibration curve must be within 25% o f the target concentration for PFOSA and PFOSAA to qualify as the LLOQ and within 20% of the target concentration to qualify as the LLOQ for all other analytes. If this criterion is not met, the next level is subjected to the same test and the LLOQ raised accordingly. Quality Control Samples Each analytical run will include low, medium and high QC samples in duplicate. The measured concentrations o f at least two-thirds o f all analytical QCs must be within 20% o f their target concentrations ( 25% for PFOSA and PFOSAA), and no two QCs at the same concentration can be outside the limit. If study samples require dilution, a dilution QC will be analyzed in triplicate for each dilution level (except for PFOSA and PFOSAA which should not be diluted with control matrix). At least two dilution QCs at each level must be within 20% o f its target concentrations in order to accept diluted study samples at that level. The dilution QC acceptance is independent o f the undiluted analytical QC acceptance. 4. RESULTS AND DISCUSSION Serum sample results obtained using plasma curves for PFOA, PFHS, PFOSAA, PFOSA, M556, and M570 did not meet NWB SOP requirements for validation. Results may vary on average up to 26% from results obtained using human serum calibration curves at some concentration levels for PFOA, PFHS, PFOSAA, M 556, and M570. For PFOSA, results may vary on average up to 43% from results obtained using human serum calibration curves. These data are reported in Assay Revalidation Addendum Report NWBR00-122 [5.3]. However, in order to obtain the lower limit of quantitation o f less than 5 ppb required by the sponsor, it was necessary to use plasma calibration curves. Percent theoretical and percent bias are separate calculations determined from the mean. Therefore, due to rounding, percent theoretical plus percent bias may not equal 100.0%. Page 13 Northwest Bioanalytical Study No. NWBSOO-145 Report No. NWBRO1-155 Each accepted run met the acceptance criteria set for the calibration curve points, for the lower limit o f quantitation (LLOQ) and for the analytical quality control (QC) samples. Run PFOS ID Result PFOA Result PFIIS PFOSAA Result Result l Accepted Accepted Accepted Accepted 2 Accepted Accepted Accepted Accepted Comments 3 Accepted Accepted Accepted Accepted 4 N/A N/A N/A N/A Run aborted, sample preparation error 5 Accepted Accepted Accepted Accepted 6 Accepted Accepted Accepted Accepted 7 Accepted Accepted Accepted Accepted 8 Accepted Accepted Accepted Accepted N/A = not applicable Run PFOSA ID Result 1 Accepted 2 Accepted 3 Accepted 4 N/A 5 Accepted 6 Accepted 7 Accepted 8 Accepted M556 Result Accepted Accepted Accepted N/A Accepted Accepted Accepted Accepted M570 Result Accepted Accepted Accepted N/A Accepted Accepted Accepted Accepted Comments Run aborted, sample preparation error N/A ==not applicable Any known circumstances that may have affected the quality or integrity o f the data are included in this report. 5. REFERENCES 5.1. 3M Company Protocol EPI-00I6. "Identification o f Fluorochemicals in Serum from and Elderly Population in the Seattle, W ashington Metropolitan Area." Page 14 Northwest Bioanalytieal Study No. NWBS00-145 Report No. NWBROI-155 5.2. C. Sakashita. "Quantitative Determination o f PFOS, PFOSA, PFOSAA, POAA, PFHS, M556 and M570 in Human Serum by LC/MS/MS." Assay Revalidation Report. NWB Study NWBS00-040. NWB Report NWBR00-108. January 24, 2001. 5.3. C. Sakashita. "Quantitative Determination o f PFOS, PFOSA, PFOSAA, POAA, PFHS, M556 and M570 in Human Serum by LC/MS/MS." Assay Revalidation Addendum Report. NWB Study NW BS00-040. NWB Report NW BR00-122. November 20, 2001. 6. DATA RETENTION The raw data and final report for this study will be stored in the NWB Archives, 1121 East 3900 South, Salt Lake City, UT 84124 per regulations and contract agreement. After submission o f the final report to the Sponsor, remaining study samples will be stored under required conditions until confirmation o f Sample Disposition/Retum Authorization is received from the Sponsor. 3M Company will be notified concerning final disposition of records at completion of contract obligations. Page 15 Northwest Bioanalytical Study No. NWBSOO-145 Report No. NWBR01-155 Table 1. Calibration Curve Summary for PFOS Quadratic weighted l/x2. All concentrations are expressed as ppb. Run Date Run Number A B O8-N0V-2 OOI 1 0.000000 0.004611 O9-N0V-2 OOI 2 0.000000 0.004551 I3 -N0V-2 OOI 3 -0.000001 0.004805 I4 -N0V-2 OOI 5 0.000000 0.005168 I5 -N0V-2OOI 6 -0.000001 0.004975 I5-N0V-2OOI 7 -0.000001 0.004756 I6 -N0V-2OOI 8 0.000000 0.005736 C 0.001070 -0.000952 0.001604 0.001523 -0.001202 -0.001763 -0.000223 RSquared 0.9984 0.9958 0.9974 0.9917 0.9985 0.9956 0.9964 LLOQ ULOQ 3.37 413 3.37 413 3.37 413 3.37 413 3.37 413 3.37 413 3.37 413 Mean S.D. %CV n 0.000000 0.004943 0.000001 0.000408 8.3 77 0.000008 0.001387 17337.5 7 0.9963 0.0023 0.2 7 A, B, and C are coefficients used to define the calibration curve. Page 16 Northwest Bioanalytieal Study No. NWBS00-145 Report No. NWBROl-155 Table 2. Calibration Curve Summary for PFOA Quadratic weighted 1/x2. All concentrations are expressed as ppb. Run Date O8-N0V-2OOI O9-N0V-2OOI I3-N0V-2OOI 14-Nov-2001 15-Nov-2001 15-Nov-2001 I6-N0V-2OOI Run A B C R- Number Squared 1 0.000001 0.006091 0.001738 0.9965 2 0.000001 0.005194 O.OOO8O8 0.9931 3 0.000000 0.005054 0.000831 0.9960 5 -0.000001 0.005711 0.000382 0.9966 6 0.000001 0.006132 -0.000212 0.9961 7 0.000001 0.006102 0.000361 0.9977 8 0.000000 0.006771 -0.000296 0.9970 LLOQ 1.44 1.44 1.44 1.44 1.44 1.44 1.44 ULOQ 481 481 481 481 481 481 481 Mean S.D. %CV n 0.000000 0.005865 0.000516 0.000001 0.000596 0.000696 10.2 134.9 777 0.9961 0.0015 0.2 7 A, B, and C are coefficients used to define the calibration curve. Page 17 Northwest Bioanalytical Study No. NWBS00-145 Report No. NWBRO1-155 Table 3. Calibration Curve Summary for PFHS Quadratic weighted 1/x2. All concentrations are expressed as ppb. Ruu Date Run A B C Number O8-N 0 V-2 OOI 1 -0.000002 0.007829 0.000656 O9 -N0 V-2OOI 2 -0.000002 0.008314 -0.001142 I3-N 0 V-2OOI 3 -0.000001 0.005315 0.000115 I4 -N 0 V-2 OOI 5 -0.000001 0.006211 -0.000433 I 5-N 0V-2 OOI 6 -0.000002 0.006849 -0.001305 I5 -N 0V-2 OOI 7 -0.000002 0.006791 -0.000751 I6-N 0V-2 OOI 8 -0.000002 0.006953 0.000081 R- LLOQ ULOQ Squared 0.9971 1.36 522 0.9969 1.36 522 0.9972 1.36 522 0.9960 1.36 522 0.9977 1.36 522 0.9978 1.36 522 0.9986 1.36 522 Mean S.D. %CV n -0.000002 0.006895 -0.000397 0.000000 0.000989 0.000719 0.0 14.3 -181.1 77 7 0.9973 0.0008 0.1 7 A, B, and C are coefficients used to define the calibration curve. Page 18 Northwest Bioanalytical Study No. NWBSOO-145 Report No. NWBROl-155 Table 4. Calibration Curve Summary for PFOSAA Quadratic weighted 1/x2. All concentrations are expressed as ppb. Run Date Run Number A O8-N 0V-2 OOI 1 0.000002 O9-N 0 V-2 OOI 2 0.000001 I3 -N 0V-2 OOI 3 -0.000001 14-Nov-2001 5 -0.000001 I5-N 0V-2 OOI 6 0.000001 I5 -N 0V-2OOI 7 0.000002 I6-N 0V-2 OOI 8 0.000003 B C R- LLOQ Squared 0.004883 0.002297 0.005152 0.000251 0.9868 0.9916 1.50 1.50 0.010855 -0.001647 0.010710 -0.000618 0.008626 -0.000767 0.9963 0.9811 0.9955 1.50 1.50 1.50 0.007392 -0.001672 0.9905 1.50 0.011058 -0.000963 0.9960 1.50 ULOQ 501 501 501 501 501 501 501 - Mean S.D. %CV n 0.000001 0.000002 200.0 7 0.008382 -0.000446 0.002659 0.001376 31.7 -308.5 77 0.9911 0.0056 0.6 7 A, B, and C are coefficients used to define the calibration curve. Page 19 Northwest Bioanalytical Study No. NWBS00-145 Report No. NWBRO1-155 Table 5. Calibration Curve Summary for PFOSA Quadratic weighted l/x2. All concentrations are expressed as ppb. Run Date Run Number A O8-N0 V-2OOI 1 0.000001 O9 -N0 V-2 OOI 2 0.000001 I3-N 0 V-2 OOI 3 -0.000002 I4 -N0 V-2OOI 5 -0.000001 I5-N0 V-2 OOI 6 0.000002 I5 -N0 V-2 OOI 7 0.000002 I6-N0V-2 OOI 8 0.000004 B 0.005637 0.005563 0.014503 0.013789 0.010415 0.010693 0.014421 C R- LLOQ ULOQ Squared 0.000591 0.9955 1.00 500 0.001213 0.9908 1.00 500 0.002158 0.9896 1.00 500 0.002219 0.9880 1.00 500 0.000499 0.9971 1.00 500 0.000700 0.9940 1.00 500 0.003092 0.9962 1.00 500 Mean S.D. %CV n 0.000001 0.000002 200.0 7 0.010717 0.003872 36.1 7 0.001496 0.001003 67.0 7 0.9930 0.0036 0.4 7 A, B, and C are coefficients used to define the calibration curve. Page 20 Northwest Bioanalytical Study No. NWBSOO-145 Report No. NWBROl-155 Tabic 6. Calibration Curve Summary for M556 Quadratic weighted 1/x2. All concentrations are expressed as ppb. Run Date 08-Nov-2001 09-NOV-2001 13- N o v -2 0 0 1 14-NOV-2001 15-Nov-2001 15-NOV-2001 16-NOV-2001 Run Number 1 2 3 5 6 7 8 A 0.000001 0.000001 0.000000 0.000000 0.000001 0.000002 0.000001 B 0.006345 0.006891 0.004174 0.003962 0.003844 0.003291 0.004646 C R- LLOQ ULOQ Squared -0.000642 0.9952 2.50 500 -0.001177 0.9950 2.50 500 -0.002780 0.9942 2.50 500 0.001886 0.9844 2.50 500 -0.002125 0.9986 2.50 500 -0.000626 0.9947 2.50 500 -0.001463 0.9970 2.50 500 Mean S.D. %CV n 0.000001 0.000001 100.0 7 0.004736 0.001356 28.6 7 -0.000990 0.001488 -150.3 7 0.9942 0.0046 0.5 7 A, B, and C are coefficients used to define the calibration curve. Page 21 Northwest Bioanalytical Study No. NWBS00-145 Report No. NWBROM55 Table 7. Calibration Curve Summary for M570 Quadratic weighted 1/x". All concentrations are expressed as ppb. Run Date Run N um ber O8-N0 V-2OOI 1 O9 -N 0 V-2 OOI 2 I3 -N 0 V-2 OOI 3 I4 -N 0 V-2OOI 5 I5 -N 0V-2 OOI 6 I5 -N 0 V-2 OOI 7 I 6-N 0 V-2 OOI 8 A 0.000001 0.000001 -0.000003 -0.000002 0.000001 0.000001 0.000001 B 0.007316 0.007844 0.015001 0.014938 0.013060 0.011552 0.016317 C 0.000681 0.001194 0.000743 0.001637 0.001232 0.000705 0.002517 R- LLOQ ULOQ Squared 0.9969 1.00 500 0.9924 1.00 500 0.9964 1.00 500 0.9922 1.00 500 0.9940 1.00 500 0.9936 1.00 500 0.9979 1.00 500 Mean S.D. %CV n 0.000000 0.000002 7 0.012290 0.003564 29.0 7 0.001244 0.000663 53.3 7 0.9948 0.0023 0.2 7 A, B, and C are coefficients used to define the calibration curve. Page 22 Northwest Bioanalytieal Study No. NWBSOO-145 Report No. NWBROI-155 Table 8. Back-Calculated Concentrations of Calibration Standards for PFOS Quadratic weighted 1/x2. All concentrations are expressed as ppb. Run Date Run 3.37 4.60 10.8 23.1 43.6 84.6 208 331 413 Number O8-N0V-2OOI 1 3.44 4.54 10.9 23.0 43.8 84.8 212 326 424 3.42 4.37 10.9 24.4 43.3 77.8 213 342 393 O9-N0V-2OOI 2 3.58 4.49 11.1 22.9 42.2 87.3 219 336 373 3.37 4.39 10.4 21.7 41.9 85.6 238 335 403 I3-N0V-2OOI 3 3.57 4.80 10.8 24.9 44.1 84.8 220 338 399 3.16 4.50 10.3 22.4 41.6 81.8 215 324 410 14-NOV-2001 5 3.07 5.33 10.1 23.8 43.6 82.5 213 317 412 2.97 5.41 10.2 22.7 42.8 83.6 217 334 416 15-N0V-2OOI 6 3.46 4.77 10.8 24.1 40.7 85.5 208 338 426 3.24 4.51 10.7 24.1 42.0 86.3 205 325 402 I 5-N0V-2 OOI 7 3.06 4.58 10.4 23.1 41.5 77.2 209 314 443 3.43 5.09 10.9 24.8 45.4 83.7 217 323 407 I6-N0V-2OOI 8 3.63 4.32 11.6 23.8 40.6 80.8 209 328 403 3.21 4.41 11.4 24.2 44.7 78.3 206 342 425 Mean S.D. %CV %Bias n 3.33 4.68 10.8 23.6 42.7 82.9 214 330 410 0.211 0.357 0.440 0.947 1.47 3.27 8.28 8.98 17.0 6.3 7.6 4.1 4.0 3.4 3.9 3.9 2.7 4.1 -1.2 1.7 0.0 2.2 -2.1 -2.0 2.9 -0.3 -0.7 14 14 14 14 14 14 14 14 14 Page 23 Northwest Bioanalytieal Study No. NWBS00-145 Report No. NWBROL-155 Table 9. Back-Calculatcd Concentrations of Calibration Standards for PFOA Quadratic weighted l/x2. All concentrations are expressed as ppb. Run Date Run 1.44 2.88 10.1 24.5 48.5 97.1 241 385 481 Number O8-N0V-2OOI 1 1.56 3.08 10.1 24.5 49.1 92.3 248 373 470 1.31 2.73 9.56 26.2 49.4 89.7 255 403 476 09-Nov-2001 2 1.71 2.73 10.2 25.1 50.4 103 251 388 444 1.26 2.73 9.55 22.5 47.2 97.5 266 390 472 I3-N0V-2OOI 3 1.60 2.90 9.74 23.8 47.7 91.7 245 377 443 1.27 2.95 9.74 26.2 50.1 96.7 252 414 488 I4 -N0V-2 OOI 5 1.44 *4.47 9.53 27.5 48.7 99.6 229 380 484 1.47 *5.21 8.81 24.9 49.4 95.7 250 401 465 I5 -N0V-2 OOI 6 1.36 2.58 9.29 24.7 43.9 101 244 375 481 1.56 2.97 11.1 25.5 48.3 99.0 247 391 475 I5 -N0 V-2 OOI 7 1.58 2.90 9.72 25.0 48.9 91.9 246 385 489 1.33 2.77 9.74 26.1 50.8 95.6 238 389 470 I6 -N0V-2 OOI 8 1.54 3.12 9.94 25.5 49.2 93.0 233 390 459 1.30 2.80 9.91 25.5 48.5 93.5 233 408 496 Mean S.D. %CV %Bias n 1.45 2.86 9.78 25.2 48.7 95.7 246 390 472 0.145 0.160 0.513 1.19 1.69 3.94 9.83 12.4 15.7 10.0 5.6 5.2 4.7 - 3.5 4.1 4.0 3.2 3.3 0.7 -0.7 -3.2 2.9 0.4 -1.4 2.1 1.3 -1.9 14 12 14 14 14 14 14 14 14 * Sample deactivated as an outlier (not included in summary statistics) Page 24 Northwest Bioanalytical Study No. NWBS00-145 Report No. NWBR01-I55 Table 10. Back-Calculated Concentrations of Calibration Standards for PFHS Quadratic weighted l/x2. All concentrations are expressed as ppb. Run Date Run 1.36 2.92 10.7 26.4 52.5 104 261 417 522 Number O8-N 0 V-2 OOI 1 1.44 2.94 11.5 29.2 53.3 99.8 264 412 540 1.31 2.72 10.3 26.6 49.3 99.4 260 420 515 O9-N0V-2OOI 2 1.43 2.97 11.2 27.3 53.6 111 282 448 499 1.33 2.74 10.2 25.2 49.8 98.2 271 398 498 I 3 -N 0V-2 OOI 3 1.40 2.95 10.1 30.1 51.5 100 262 420 509 1.31 2.97 10.6 25.5 50.9 103 278 401 536 I4 -N 0V-2 OOI 5 1.39 *4.07 10.6 28.5 51.2 93.9 304 408 521 1.33 *4.42 11.1 26.6 49.7 100 262 398 523 I5-N0V-2OOI 6 1.28 2.93 11.2 26.9 52.7 105 263 436 533 1.46 2.78 11.4 26.1 48.9 99.0 259 409 507 I5 -N 0V-2 OOI 7 1.39 2.76 10.1 26.3 52.4 106 253 394 516 1.42 2.71 10.7 28.5 55.7 104 265 428 538 I6-N0V-2OOI 8 1.36 3.03 10.6 26.7 49.8 103 264 421 508 1.34 2.85 10.9 28.6 51.9 97.8 258 426 532 Mean S.D. %CV %Bias n 1.37 2.86 10.8 27.3 51.5 101 268 416 520 0.0557 0.115 0.477 1.46 1.94 4.24 13.0 15.7 14.4 4.1 4.0 4.4 5.3 3.8 4.2 4.9 3.8 2.8 0.7 -2.1 0.9 3.4 -1.9 -2.9 2.7 -0.2 -0.4 14 12 14 14 14 14 14 14 14 * Sample deactivated as an outlier (not included in summary statistics) Page 25 Northwest Bioanalytical Study No. NWBS00-145 Report No. NWBRO1-155 T able 11. B ack-C alculated C oncentrations o f C alibration S tandards for PFOSAA Quadratic weighted 1/x2. All concentrations are expressed as ppb. Run Date Run 1.50 3.00 10.5 25.5 50.5 101 251 401 501 Number 08-NOV-2001 1 09-NOV-2001 2 13-NOV-2001 3 14-NOV-2001 5 15-NOV-2001 6 15-NOV-2001 7 16-NOV-2001 8 1.23 2.63 10.3 25.2 52.8 111 276 428 509 1.90 2.87 10.3 26.5 51.0 86.7 251 399 448 1.62 2.88 10.8 26.6 53.8 105 266 405 451 1.51 2.63 9.67 23.7 46.8 98.0 300 416 473 1.61 3.00 11.0 24.9 54.1 106 266 435 457 1.41 2.94 9.72 24.7 49.7 95.7 243 415 488 1.40 *4.70 8.98 27.6 48.3 102 207 386 515 1.38 4.10 8.98 26.7 52.3 99.2 269 442 489 1.49 3.11 10.4 28.9 42.5 103 261 411 525 1.49 2.95 10.6 26.8 48.9 98.9 235 401 477 1.49 2.67 9.62 24.0 48.9 93.3 245 361 564 1.48 3.52 10.4 27.7 53.6 108 275 376 482 1.53 2.76 10.6 28.1 53.5 101 255 411 506 1.62 2.61 10.2 25.8 52.3 91.9 251 398 488 4 Mean S.D. %CV %Bias n 1.51 2.97 10.1 26.2 50.6 100 257 406 491 0.153 0.420 0.629 1.57 3.31 6.63 21.9 22.0 31.4 10.1 14.1 6.2 6.0 6.5 6.6 V"> od 5.4 6.4 0.7 -1.0 -3.8 2.7 0.2 -1.0 2.4 1.2 -2.0 14 13 14 14 14 14 14 14 14 * Sample deactivated as an outlier (not included in summary statistics) Page 26 Northwest Bioanalytieal Study No. NWBSOO-145 Report No. NWBRO1-155 Table 12. Back-Calculated Concentrations of Calibration Standards for PFOSA Quadratic weighted lIk2. All concentrations are expressed as ppb. Run Date Run 1.00 2.50 10.0 25.0 50.0 100 250 400 500 Number 08-NOV-2001 1 0.955 2.45 9.66 23.7 51.9 100 266 396 511 0.992 2.91 10.0 26.0 48.4 87.5 253 413 472 09-Nov-2001 2 0.831 2.61 9.84 25.5 51.9 100 263 376 487 1.19 2.33 9.56 23.1 48.6 98.4 298 403 479 13-Nov-2001 3 1.09 2.89 10.6 25.0 56.8 108 258 446 462 0.875 2.37 8.69 23.2 46.8 91.4 231 423 480 14-NOV-2001 5 1.00 *4.45 ' 8.89 27.4 48.0 98.3 208 395 496 0.919 3.12 8.63 26.8 52.2 94.6 268 436 502 15-NOV-2001 6 1.09 2.66 10.2 25.7 49.2 100 256 400 525 0.919 2.27 10.2 25.9 47.4 100 236 406 477 15-NOV-2001 7 1.04 2.45 8.98 25.0 50.5 89.4 236 423 527 0.934 2.79 9.85 27.1 54.0 94.2 272 385 466 16-NOV-2001 8 1.10 2.66 9.95 27.6 50.7 94.7 249 397 510 0.915 2.24 9.88 25.7 51.1 94.1 237 418 494 Mean S.D. %CV %Bias n 0.989 2.60 9.64 25.6 50.5 96.5 252 408 492 0.101 0.274 0.610 1.45 2.75 5.29 22.0 19.4 20.9 10.2 10.5 6.3 5.7 5.4 5.5 8.7 4.8 4.2 -1.1 4.0 -3.6 2.4 1.0 -3.5 0.8 2.0 -1.6 14 13 14 14 14 14 14 14 14 * Sample deactivated as an outlier (not included in summary statistics) Page 27 Northwest Bioanalytical Study No. NWBS00-I45 Report No. NWBROl-155 Table 13. Back-Calculated Concentrations of Calibration Standards for M556 Quadratic weighted 1/x2. All concentrations are expressed as ppb. Run Date Run 2.50 10.0 25.0 50.0 100 250 400 500 Number O8-N 0V-2 OOI 1 2.26 9.38 23.4 51.9 103 259 410 507 2.75 10.3 27.3 50.5 87.1 255 405 472 O9 -N 0V-2 OOI 2 2.65 10.1 25.8 50.2 99.3 261 389 456 2.36 9.91 23.1 46.8 97.5 291 416 494 I3-N0V-2OOI 3 2.59 11.1 25.2 53.6 102 276 430 476 2.43 8.53 24.1 49.7 92.5 238 404 479 M-Nov-2001 5 *4.38 8.25 30.4 51.8 107 224 396 490 2.70 7.51 23.5 54.9 101 268 431 476 I5 -N0V-2 OOI 6 2.48 10.1 24.7 45.5 100 257 407 515 2.52 10.0 25.1 51.0 106 252 387 480 I5 -N0V-2 OOI 7 2.31 9.28 25.4 50.5 88.4 251 390 539 2.72 10.0 24.8 54.2 103 268 388 467 I6-N0V-2OOI 8 2.61 9.97 26.5 46.2 102 245 401 496 2.34 10.5 26.9 45.1 97.5 252 421 490 Mean S.D. %CV %Bias n 2.52 9.64 25.4 50.1 99.0 257 405 488 0.167 0.964 1.92 3.18 - 6.01 16.3 14.9 21.4 6.6 10.0 7.6 6.3 6.1 6.3 3.7 4.4 0.8 -3.6 1.6 0.2 - 1.0 2.8 1.3 -2.4 13 14 14 14 14 14 14 14 * Sample deactivated as an outlier (not included in summary statistics) Page 28 Northwest Bioanalytical Study No. NWBSOO-145 Report No. NWBRO1-155 Table 14. Back-Calculated Concentrations of Calibration Standards for M570 Quadratic weighted 1/x2. AH concentrations are expressed as ppb. Run Date Run 1.00 2.50 10. 0 25.0 50.0 100 250 400 500 Number 08-NOV-2001 1 1.03 2.47 9.84 23.1 50.9 108 263 417 511 0.987 2.42 10.2 26.3 50.2 89.3 254 397 462 09-NOV-2001 2 1.11 2.52 10.1 25.9 51.4 104 264 403 443 0.925 2.27 9.96 23.2 45.4 95.9 299 411 487 13-NOV-2001 3 1.08 2.64 10.0 24.1 52.3 102 264 421 450 0.896 2.58 9.38 24.0 49.6 96.1 255 425 484 14-NOV-2001 5 0.985 *4.20 9.07 28.9 47.3 103 212 399 506 1.03 *3.85 8.56 27.0 52.2 99.8 275 424 479 15-NOV-2001 6 1.10 2.64 9.91 27.3 41.3 100 271 408 514 0.906 2.32 10.1 26.8 48.2 102 243 394 475 15-NOV-2001 7 0.930 2.33 9.10 25.1 49.4 90.7 239 373 561 1.07 2.72 9.95 26.7 52.6 105 276 382 470 16-NOV-2001 8 1.06 2.57 9.97 26.8 49.7 98.8 250 409 506 0.963 2.28 10.1 25.3 51.9 92.0 244 406 489 Mean S.D. %CV %Bias n 1.01 2.48 9.73 25.8 49.5 99.0 258 0.0736 0.155 0.499 1.69 3.13 5.60 20.7 7.3 6.3 5.1 6.6 6.3 5.7 8.0 1.0 -0.8 -2.7 3.2 -1.0 -1.0 3.2 14 12 14 14 14 14 14 * Sample deactivated as an outlier (not included in summary statistics) 405 488 15.2 30.3 3.8 6.2 1.3 -2.4 14 14 Page 29 Northwest Bioanalytical Study No. NWBS00-145 Report No. NWBROl-155 Table 15. Analytical QC Summary for PFOS All concentrations are expressed as ppb. Run Date Run Number Preparation Date O8-N0 V-2 OOI 1 O9-N0 V-2 OOI 2 I3-N 0V-2 OOI 3 M-Nov-2001 5 I5-N0 V-2 OOI 6 I5 -N0 V-2 OOI 7 I6-N0 V-2 OOI 8 Low QC 5.83 ppb O5 -N0 V-2 OOI 5.89 5.75 6.63 6.50 6.21 6.28 5.48 5.95 6.16 6.20 5.56 6.11 6.21 6.17 Medium QC 126 ppb O5 -N0 V-2 OOI 121 127 120 121 134 118 108 114 128 118 114 117 124 120 High QC 331 ppb O5 -N 0 V-2 OOI 336 349 338 333 363 335 303 322 338 347 323 355 330 323 Mean S.D. %CV %Theoretical %Bias n 6.08 0.325 5.3 104.3 4.3 14 120 6.56 5.5 95.2 -4.8 14 335 15.3 4.6 101.2 1.2 14 Page 30 Northwest Bioanalytical Study No. NWBSOO-145 Report No. NWBR01-155 Table 16. Analytical QC Summary for PFOA All concentrations are expressed as ppb. Run Date Run Number Low QC 4.32 ppb M edium QC 144 ppb High QC 384 ppb Preparation Date 05-NOV-2001 05-Nov-2001 05-NOV-2001 08-NOV-2001 1 4.04 138 384 3.80 141 395 09-NOV-2001 '2 4.25 137 391 3.78 138 382 13-NOV-2001 3 4.03 156 436 4.46 141 430 14-Nov-2001 5 4.16 4.63 132. 351 135 391 15-Nov-2001 6 4.30 139 394 4.30 136 396 15-NOV-2001 7 4.25 140 379 4.65 139 399 16-NOV-2001 8 4.30 142 383 4.33 131 374 Mean S.D. %CV %Theoretical %Bias n 4.23 . 0.261 6.2 97.9 -2.1 14 139 5.89 4.2 96.5 -3.5 14 392 21.3 5.4 102.1 2.1 14 J Page 31 Northwest Bioanalytical Study No. NWBS00-145 Report No. NWB RO l-15 5 Table 17. Analytical QC Summary for PFHS All concentrations are expressed as ppb. Run Date Run Number Low QC 4.48 ppb M edium QC 156 ppb High QC 417 ppb Preparation Date O5 -N0 V-2 OOI O5-N 0 V-2 OOI O5-N0 V-2 OOI 08-NOV-2001 1 4.50 140 404 4.13 146 409 09-NOV-2001 2 4.55 140 401 4.61 142 389 13-NOV-2001 3 *6.21 176 443 4.42 144 433 14-NOV-2001 5 4.82 131 358 4.15 143 385 15-Nov-2001 6 4.58 144 397 4.27 129 408 15-Nov-2001 7 4.24 143 407 4.51 149 415 I6 -N 0 V-2 OOI 8 4.49 141 407 4.39 144 392 Mean S.D. %CV %Theoretical %Bias n 4.57 0.526 11.5 102.0 2.0 14 144 10.7 7.4 92.3 -7.7 14 * > 20% deviation from theoretical 403 20.5 5.1 96.6 -3.4 14 Page 32 Northwest Bioanalytical Study No. NWBS00-145 Report No. NWBROl-155 Table 18. Analytical QC Summary for PFOSAA All concentrations are expressed as ppb. Run Date Run Number Low QC 4.50 ppb Medium QC 151 ppb High QC 401 ppb Preparation Date O5-N 0 V-2 OOI O5-N 0 V-2OOI O5-N 0 V-2 OOI 08-NOV-2001 1 5.01 151 392 4.35 149 419 09-N o v-2001 2 5.06 149 400 4.31 145 401 13-NOV-2001 3 *3.34 4.79 150 149 446 453 14-NOV-2001 5 3.95 139 399 4.75 148 421 15-Nov-2001 6 5.03 159 443 4.32 149 398 15-NOV-2001 7 4.75 139 353 4.60 129 395 I6-N0 V-2 OOI 8 4.74 150 390 4.89 146 387 Mean S.D. %CV %Theoretical %Bias n 4.56 0.478 10.5 101.3 1.3 14 147 7.07 4.8 97.4 -2.6 14 * > 25% deviation from theoretical 407 27.0 6.6 101.5 1.5 14 Page 33 Northwest Bioanalytical Study No. NWBS00-145 Report'No. NWBROt-155 Table 19. Analytical QC Summary for PFOSA All concentrations are expressed as ppb. Run Date Run Number Preparation Date O8-N 0 V-2 OOI 1 O9-N 0 V-2 OOI 2 I3-N 0V-2 OOI 3 14-NOV-2001 5 I5-N 0 V-2OOI 6 I5 -N 0 V-2 OOI 7 I6-N 0V-2 OOI 8 Low QC 4.00 ppb M edium QC High QC 150 ppb 400 ppb O5-N 0 V-2 OOI 0 5 -Nov-2001 05-Nov-2001 3.84 149 398 4.16 149 424 4.17 133 393 3.67 133 404 3.09 159 446 4.22 153 427 3.37 140 411 4.42 139 443 4.73 151 412 3.80 142 395 3.82 131 346 3.80 121 399 4.15 142 347 3.77 135 359 Mean S.D. %CV %Theoretical %Bias n 3.93 141 400 0.420 10.3 31.7 10.7 7.3 7.9 98.3 94.0 100.0 -1.8 -6.0 0.0 14 14 14 Page 34 Northwest Bioanalytical Study No. NWBS00-I45 Report No. NWBRO1-155 Table 20. Analytical QC Summary for M556 All concentrations are expressed as ppb. Run Date Run Number Preparation Date 08-NOV-2001 L 09-NOV-2001 2 13-Nov-2001 3 14-NOV-2001 5 15-NOV-2001 6 15-Nov-2001 7 16-Nov-2001 8 Low QC 4.00 ppb Medium QC 150 ppb 05-NOV-2001 05-NOV-2001 3.80 140 4.02 145 4.58 145 4.18 143 *3.07 151 3.41 146 3.43 148 3.38 143 *5.10 157 3.95 148 3.91 148 4.16 138 4.53 145 4.53 143 High QC 400 ppb 05-NOV-2001 396 432 408 414 470 436 383 419 414 416 387 419 404 387 Mean S.D. %CV %Theoretical %Bias n 4.00 0.564 14.1 100.0 0.0 14 146 4.70 3.2 97.3 -2.7 14 * > 20% deviation from theoretical 413 23.0 5.6 103.3 3.3 14 Page 35 Northwest Bioanalytical Study No. NWBSOO-145 Report No. NWBR01-155 Table 21. Analytical QC Summary for M570 All concentrations are expressed as ppb. Run Date Run Number Low QC 4.00 ppb Medium QC 150 ppb High QC 400 ppb Preparation Date O5-N 0 V-2 OOI O5-N 0 V-2 OOI O5 -N0 V-2 OOI 08-NOV-2001 1 4.07 149 397 3.86 146 433 09-NOV-2001 2 4.55 146 405 4.20 140 404 13-NOV-2001 3 *2.95 4.32 143 149 451 455 14-NOV-2001 5 3.83 142 388 4.49 146 429 15-Nov-2001 6 4.49 153 432 3.74 147 411 15-Nov-2001 7 4.02 141 367 4.22 132 399 I6 -N 0 V-2 OOI 8 4.25 149 403 4.32 144 391 Mean S.D. %CV % T heoretical %Bias n 4.09 0.415 10.1 102.3 2.3 14 145 5.13 3.5 96.7 -3.3 14 * > 20% deviation from theoretical 412 25.0 6.1 103.0 3.0 14 Page 36 Northwest Bioanalytical Study No. NWBSOO-145 Report No. NWBR01-155 Table 22. Scrum Study Sample Concentrations All concentrations are expressed as ppb. Sample Name 11331 11332 11333 11334 11335 11336 11337 11338 11339 11340 11341 11342 11343 11344 11345 11346 11347 11348 11349 11350 11351 11352 11353 11354 11355 11356 11357 11358 11359 11360 11361 11362 11363 11364 11365 11366 PFOS 33.7 28.8 44.8 12.4 18.5 25.4 29.8 33.2 32.3 37.4 35.3 83.2 30.8 61.9 54.9 34.6 58.9 57.7 14.5 24.5 14.7 13.5 22.1 71.3 48.3 59.7 18.8 25.2 49.7 19.0 16.3 95.2 133 49.1 104 39.6 PFOSA* PFOSAA* PFOA* PFHS* M556* M570* <LLOQ(l.OO) 1.96 6.20 <LLOQ (1.36) <LLOQ (2.50) 1.30 <LLOQ(1.00) <LLOQ (1.50) 4.73 4.50 <LLOQ (2.50) 1.37 <LLOQ(1.00) 3.33 6.16 3.99 <LLOQ (2.50) 2.22 <LLOQ (1.00) 1.64 2.47 <LLOQ (1.36) <LLOQ (2.50) 1.82 <LLOQ (1.00) <LLOQ (1.50) <LLOQ (1.44) <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.00) <LLOQ ( 1.50) 2.93 2.67 <LLOQ (2.50) 1.36 <LLOQ (1.00) <LLOQ (1.50) 3.56 3.23 <LLOQ (2.50) <LLOQ (1.00) <LLOQ(I.00) <LLOQ ( 1.50) 4.03 1.73 <LLOQ (2.50) 1.72 <LLOQ ( 1.00) <LLOQ (1.50) 4.67 2.48 <LLOQ (2.50) 1.29 <LLOQ (1.00) 2.41 3.96 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.00) . 1.50 2.94 7.19 <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.00) 19.0 4.69 3.20 4.79 3.25 <LLOQ (1.00) <LLOQ (1.50) 3.78 2.00 <LLOQ (2.50) <LLOQ (1.00) <LLOQ ( 1.00) 10.7 7.35 3.35 3.04 2.68 <LLOQ (1.00) 1.68 11.4 5.00 <LLOQ (2.50) 2.50 <LLOQ (1.00) <LLOQ (1.50) 4.66 1.92 <LLOQ (2.50) 1.18 <LLOQ (1.00) 5.43 7.52 7.78 <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.00) 2.44 5.01 5.80 <LLOQ (2.50) 1.18 <LLOQ (1.00) <LLOQ (1.50) 2.84 <LLOQ (1.36) <LLOQ (2.50) 1.43 <LLOQ (1.00) <LLOQ (1.50) 4.12 1.77 <LLOQ (2.50) 1.38 <LLOQ (1.00) <LLOQ (1.50) 10.6 <LLOQ (1.36) <LLOQ (2.50) 3.55 <LLOQ (1.00) 4.73 2.37 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.00) 2.48 3.75 5.26 <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.00) 3.80 12.5 15.9 <LLOQ (2.50) 2.47 <LLOQ(1.00) 4.88 4.01 <LLOQ (1.36) <LLOQ (2.50) 1.13 <LLOQ (1.00) <LLOQ (1.50) 9.19 2.40 <LLOQ (2.50) 1.26 <LLOQ (1.00) 2.01 4.03 2.10 <LLOQ (2.50) 1.08 <LLOQ (1.00) <LLOQ (1.50) 1.94 10.3 <LLOQ (2.50) <LLOQ(1.00) <LLOQ (1.00) 1.93 6.32 8.53 <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.00) 4.58 4.27 1.99 <LLOQ (2.50) cLLOQ(l.OO) <LLOQ (1.00) <LLOQ ( 1.50) 2.58 1.69 <LLOQ (2.50) <LLOQ ( 1.00) <LLOQ ( 1.00) 17.7 8.56 3.29 2.80 1.82 <LLOQ ( 1.00) 16.2 16.7 <LLOQ (1.36) 2.89 1.49 <LLOQ (1.00) 5.30 5.24 2.48 <LLOQ (2.50) 3.76 <LLOQ (1.00) 1.95 9.16 2.13 <LLOQ (2.50) 2.74 <LLOQ (1.00) 1.63 5.13 4.42 <LLOQ (2.50) 2.97 NOTE: The method acceptance criteria states that the measured concentration for each QC must be within 20% (25% for PFOSA and PFOSAA) of the target concentration. The statistical error associated with any individual result may exceed the QC acceptance criteria. * Serum sample results for PFOA, PFHS, PFOSAA, PFOSA, M556 and M570 obtained using plasma calibration curves may vary from results obtained using serum calibration curves (see Results and Discussion section). Page 37 Northwest Bioanalytical Study No. NWBSOO-145 Report No. NWBROI-155 Table 22. Serum Study Sample Concentrations (Continued) All concentrations are expressed as ppb. Sample Name 11367 11368 11369 11370 11371 11372 11373 11374 11375 11376 11377 11378 11379 11380 11381 11382 11383 11384 11385 11386 11387 11388 11389 11390 11391 11392 11393 11394 11395 11396 11397 11398 11399 11400 11401 PFOS 13.7 10.7 31.7 30.1 22.9 27.8 18.1 40.0 18.5 21.1 71.8 35.3 11.2 60.1 29.6 35.9 26.1 19.3 30.2 47.2 53.5 28.9 38.3 31.6 18.6 37.1 25.6 43.2 24.2 73.7 41.7 45.7 10.7 22.1 44.5 PFOSA* PFOSAA* PFOA* PFIIS* M556* M570* <LLOQ (1.00) <LLOQ (1.50) 2.20 2.31 <LLOQ (2.50) <LLOQ (1.00) <LLOQ(1.00) <LLOQ (1.50) <LLOQ (1.44) <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.00) <LLOQ (1.50) 2.37 1.63 <LLOQ (2.50) 2.80 <LLOQ (1.00) 2.13 3.64 1.45 <LLOQ (2.50) 1.05 <LLOQ (1.00) 2.25 2.75 1.79 <LLOQ (2.50) 1.09 <LLOQ (1.00) <LLOQ (1.50) 3.42 1.84 <LLOQ (2.50) 1.13 <LLOQ (1.00) 2.19 2.35 <LLOQ (1.36) <LLOQ (2.50) 1.18 <LLOQ (1.00) 13.8 6.85 2.98 <LLOQ (2.50) 3.25 <LLOQ (1.00) <LLOQ (1.50) 3.32 3.04 <LLOQ (2.50) <LLOQ (1.00) <LLOQ(1.00) <LLOQ (1.50) 1.81 1.36 <LLOQ (2.50) 1.48 <LLOQ (1.00) 2.22 6.34 17.5 <LLOQ (2.50) 1.77 <LLOQ (1.00) 3.55 6.19 2.21 <LLOQ (2.50) 2.45 <LLOQ (1.00) 1.69 2.89 <LLOQ (1.36) <LLOQ (2.50) 1.00 <LLOQ (1.00) <LLOQ (1.50) 4.18 2.39 <LLOQ (2.50) <LLOQ(1.00) <LLOQ(1.00) <LLOQ (1.50) 2.47 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.00) <LLOQ (1.50) 5.08 1.58 <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.00) 1.81 3.70 1.59 <LLOQ (2.50) 3.01 <LLOQ (1.00) <LLOQ (1.50) 2.90 1.96 <LLOQ (2.50) 2.46 <LLOQ ( 1.00) <LLOQ (1.50) 4.24 2.49 <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.00) 8.12 5.17 2.57 <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.00) 2.28 5.04 2.65 <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.00) 14.6 4.65 3.95 2.67 2.06 <LLOQ (1.00) 1.73 3.82 2.40 <LLOQ (2.50) 1.49 <LLOQ (1.00) 1.64 3.44 1.90 <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.00) <LLOQ ( 1.50) 1.86 2.72 <LLOQ (2.50) 1.27 <LLOQ (1.00) <LLOQ (1.50) 2.45 1.88 <LLOQ (2.50) 1.84 <LLOQ (1.00) <LLOQ (1.50) 3.20 1.55 <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.00) 2.62 3.98 3.63 <LLOQ (2.50) 5.69 <LLOQ (1.00) 5.16 3.47 1.87 <LLOQ (2.50) <LLOQ ( 1.00) <LLOQ (1.00) 5.62 6.92 8.28 <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.00) <LLOQ (1.50) 4.30 2.48 <LLOQ (2.50) 1.02 <LLOQ (1.00) 2.27 5.50 1.94 <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.00) <LLOQ (1.50) 2.44 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) <LLOQ ( 1.00) <LLOQ ( 1.50) 3.00 <LLOQ (1.36) <LLOQ (2.50) 1.31 <LLOQ (1.00) 3.24 5.03 5.12 <LLOQ (2.50) <LLOQ (1.00) NOTE: The method acceptance criteria states that the measured concentration for each QC must be within 20% (25% for PFOSA and PFOSAA) of the target concentration. The statistical error associated with any individual result may exceed the QC acceptance criteria. * Serum sample results for PFOA, PFHS, PFOSAA, PFOSA, M556 and M570 obtained using plasma calibration curves may vary from results obtained using serum calibration curves (see Results and Discussion section). Page 38 Northwest Bioanalytical Study No. NWBS00-145 Report No. NWBR01-155 Table 22. Scrum Study Sample Concentrations (Continued) All concentrations are expressed as ppb. Sample Name 11402 11403 11404 11405 11406 11407 11408 11409 11410 11411 11412 11413 11414 11415 11416 11417 11418 11419 11420 11421 11422 11423 11424 11425 11426 11427 11428 11429 11430 11431 11432 11433 11434 11435 11436 PFOS 51.5 21.7 29.4 22.3 40.6 40.6 27.0 13.8 104 25.1 26.5 34.1 18.0 61.3 22.1 58.9 25.6 26.3 27.2 61.3 <LLOQ (3.37) 37.3 25.4 15.1 25.8 41.6 29.3 29.7 41.1 26.3 114 48.8 30.7 24.3 14.6 PFOSA* PFOSAA* PFOA* PFHS* M556* M570* <LLOQ(1.00) <LLOQ (1.50) 6.35 9.74 <LLOQ (2.50) 1.39 <LLOQ (1.00) 2.94 4.57 1.80 <LLOQ (2.50) 2.08 <LLOQ (1.00) <LLOQ (1.50) 2.99 3.42 <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.00) 1.74 2.19 1.43 <LLOQ (2.50) 2.13 <LLOQ (1.00) <LLOQ (1.50) 3.56 4.57 <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.00) <LLOQ (1.50) 7.00 7.00 <LLOQ (2.50) 1.01 <LLOQ (1.00) 2.06 3.57 2.46 <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.00) <LLOQ (1.50) 4.17 2.22 <LLOQ (2.50) 1.13 <LLOQ (1.00) 2.57 10.2 2.33 <LLOQ (2.50) 4.05 <LLOQ (1.00) 3.33 <LLOQ (1.44) 1.71 <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.00) <LLOQ (1.50) 3.14 8.74 <LLOQ (2.50) 2.71 <LLOQ (1.00) <LLOQ (1.50) 4.44 1.40 <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.00) <LLOQ (1.50) 3.84 6.32 <LLOQ (2.50) <LLOQ (1.00) <LLOQ(1.00) 3.79 7.53 6.43 <LLOQ (2.50) <LLOQ (1.00) <LLOQ ( 1.00) <LLOQ (1.50) 2.28 <LLOQ (1.36) <LLOQ (2.50) 1.44 <LLOQ (1.00) <LLOQ (1.50) 5.67 17.1 <LLOQ (2.50) 1.83 <LLOQ (1.00) 2.43 4.69 2.78 <LLOQ (2.50) 1.14 <LLOQ (1.00) 2.23 4.30 1.57 <LLOQ (2.50) 1.62 <LLOQ (1.00) <LLOQ (1.50) 2.27 2.02 <LLOQ (2.50) 1.95 <LLOQ ( 1.00) ' 10.5 8.76 6.97 <LLOQ (2.50) 3.31 <LLOQ ( 1.00) <LLOQ (1.50) <LLOQ (1.44) <LLOQ (1.36) <LLOQ (2.50) <LLOQ ( 1.00) <LLOQ ( 1.00) <LLOQ (1.50) 4.90 3.52 <LLOQ (2.50) cLLOQ(l.OO) <LLOQ ( 1.00) 5.31 6.62 1.92 <LLOQ (2.50) 4.74 <LLOQ(1.00) <LLOQ (1.50) 3.43 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.00) <LLOQ (1.50) 2.64 6.22 <LLOQ (2.50) 3.48 <LLOQ (1.00) <LLOQ (1.50) 8.98 2.66 <LLOQ (2.50) 6.30 <LLOQ (1.00) 2.94 5.90 1.68 <LLOQ (2.50) 3.51 <LLOQ ( 1.00) <LLOQ (1.50) 4.44 , 4.74 <LLOQ (2.50) 1.63 <LLOQ ( 1.00) <LLOQ (1.50) 4.08 2.15 <LLOQ (2.50) <LLOQ (1.00) <LLOQ ( 1.00) <LLOQ (1.50) 2.56 1.70 <LLOQ (2.50) 3.71 <LLOQ ( 1.00) 4.63 14.4 2.32 <LLOQ (2.50) 3.03 <LLOQ (1.00) <LLOQ (1.50) 9.17 7.42 <LLOQ (2.50) 6.15 <LLOQ ( 1.00) 1.87 6.89 2.22 <LLOQ (2.50) 1.16 <LLOQ ( l .00) 3.14 3.72 1.59 <LLOQ (2.50) <LLOQ (1.00) <LLOQ ( l .00) <LLOQ (1.50) 2.82 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) NOTE: The method acceptance criteria states that the measured concentration for each QC must be within 20% (25% for PFOSA and PFOSAA) of the target concentration. The statistical error associated with any individual result may exceed the QC acceptance criteria. * Serum sample results for PFOA, PFHS, PFOSAA, PFOSA, M 556 and M570 obtained using plasma calibration curves may vary from results obtained using serum calibration curves (see Results and Discussion section). Page 39 Northwest Bioanalytieal Study No. NWBS00-145 Report No. NWBROI-155 Table 22. Serum Study Sample Concentrations (Continued) All concentrations are expressed as ppb. Sample Name 11437 11438 11439 11440 11441 11442 11443 11444 11445 11446 11447 11448 11449 11450 11451 11452 11453 11454 11455 11456 11457 11458 11459 11460 11461 11462 11463 11464 11465 11466 11467 11468 11469 11470 11471 11472 PFOS 20.2 32.8 49.9 15.4 20.3 12.2 34.6 17.2 11.7 29.2 20.4 13.4 36.7 36.8 50.8 39.0 34.3 29.0 27.6 16.4 138 19.0 46.1 51.6 42.9 175 9.06 26.7 17.5 33.6 32.6 64.7 28.1 22.0 61.4 28.3 PFOSA* PFOSAA* <LLOQ(1.00) 5.89 <LLOQ (1.00) <LLOQ (1.50) <LLOQ (1.00) 8.95 <LLOQ(1.00) <LLOQ ( l .50) <LLOQ (1.00) <LLOQ (1.50) <LLOQ (1.00) <LLOQ (1.50) <LLOQ (1.00) 2.26 <LLOQ (1.00) <LLOQ (1-50) <LLOQ(1.00) <LLOQ (1.50) <LLOQ (1.00) <lLOQ (1.50) <LLOQ ( 1.00) <LLOQ (1.50) <LLOQ (1.00) <LLOQ (1.50) <LLOQ (1.00) 3.76 <LLOQ (1.00) <LLOQ (1.50) <LLOQ (1.00) <LLOQ (1.50) <LLOQ (1.00) <LLOQ (1.50) <LLOQ (1.00) 2.86 <LLOQ(1.00) 2.18 <LLOQ(1.00) 1.62 <LLOQ ( 1.00) 2.40 <LLOQ(1.00) 21.1 <LLOQ (1.00) 1.57 <LLOQ (1.00) <LLOQ (1.50) <LLOQ ( 1.00) 2.81 <LLOQ (1.00) <LLOQ (1.00) <LLOQ ( 1.00) 1.60 2.46 1.79 <LLOQ (1.00) <LLOQ (1.50) <LLOQ (1.00) 2.00 <LLOQ (1.00) <LLOQ (1.50) <LLOQ ( 1.00) 1.78 <LLOQ (1.00) 2.77 <LLOQ (1.00) 1.77 <LLOQ ( 1.00) <LLOQ (1.50) <LLOQ (1.00) 6.75 <LLOQ(1.00) <LLOQ (1.50) PFOA* 2.63 6.47 5.96 3.33 4.30 1.84 4.54 2.33 1.87 3.45 2.99 2.78 4.05 3.18 4.87 2.85 4.44 4.19 2.48 3.42 11.3 2.76 4.81 6.17 2.21 9.25 4.35 3.02 2.99 4.40 5.41 6.84 3.96 1.81 9.95 4.22 PFHS* M556* M570* <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) 3.22 <LLOQ (2.50) 1.55 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) 2.05 <LLOQ (2.50) 1.28 2.15 <LLOQ (2.50) <LLOQ (1.00) 2.65 <LLOQ (2.50) <LLOQ (1.00) 4.25 <LLOQ (2.50) 1.70 <LLOQ (1.36) <LLOQ (2.50) <LLOQ ( 1.00) 3.71 <LLOQ (2.50) <LLOQ (1.00) 2.30 <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.36) <LLOQ (2.50) 1.18, 2.69 <LLOQ (2.50) 2.86 1.72 . <LLOQ (2.50) 2.58 1.80 <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) 1.47 <LLOQ (2.50) <LLOQ(1.00) <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.36) <LLOQ (2.50) 1.16 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.36) 2.82 1.22 <LLOQ (1.36) <LLOQ (2.50) 2.86 5.25 <LLOQ (2.50) <LLOQ (1.00) 1.58 <LLOQ (2.50) 1.52 5.44 <LLOQ (2.50) 4.94 17.0 <LLOQ (2.50) 1.01 4.90 <LLOQ (2.50) 1.02 2.74 <LLOQ (2.50) 1.71 <LLOQ (1.36) <LLOQ (2.50) 2.00 4.23 <LLOQ (2.50) 1.75 4.00 <LLOQ (2.50) 2.81 1.59 <LLOQ (2.50) 4.23 2.16 <LLOQ (2.50) <LLOQ(1.00) <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) 1.64 <LLOQ (2.50) 2.87 3.56 <LLOQ (2.50) <LLOQ (1.00) NOTE: The method acceptance criteria states that the measured concentration for each QC must be within 20% (25% for PFOSA and PFOSAA) of the target concentration. The statistical error associated with any individual result may exceed the QC acceptance criteria. Serum sample results for PFOA, PFHS, PFOSAA, PFOSA, M556 and M570 obtained using plasma calibration curves may vary from results obtained using serum calibration curves (see Results and Discussion section). Page 40 Northwest Bioanalytical Study No. NWBSOO-145 Report No. NWBRO1-155 Table 22. Serum Study Sample Concentrations (Continued) All concentrations are expressed as ppb. Sample Name 11473 11474 11475 11476 11477 11478 11479 11480 11481 11482 11483 11484 11485 11486 11487 11488 11489 11490 11491 11492 11493 11494 11495 11496 11497 11498 11499 11500 11501 11502 11503 11504 11505 11506 11507 11508 PFOS 32.1 27.9 27.4 18.9 38.1 47.5 28.4 48.4 26.0 19.7 47.2 72.5 33.6 31.8 37.3 25.2 21.4 29.6 91.5 50.2 24.4 47.0 48.6 47.3 76.4 34.2 57.0 20.5 17.2 42.2 56.1 30.5 104 44.0 32.1 24.3 PFOSA* PFOSAA* <LLOQ (1.00) <LLOQ (1.50) <LLOQ (1.00) <LLOQ (1.50) <LLOQ (1.00) <LLOQ (1.50) <LLOQ (1.00) <LLOQ (1.50) <LLOQ (1.00) 1.96 <LLOQ (1.00) 1.61 <LLOQ (1.00) <LLOQ (1.50) <LLOQ (1.00) 6.49 <LLOQ (1.00) <LLOQ (1.50) <LLOQ (1.00) <LLOQ (1.50) <LLOQ (1.00) 2.50 <LLOQ (1.00) 7.66 <LLOQ (1.00) <LLOQ (1.50) <LLOQ(1.00) <LLOQ (1.50) <LLOQ ( 1.00) 3.40 <LLOQ (1.00) 1.68 <LLOQ (1.00) <LLOQ (1.50) <LLOQ (1.00) 2.67 <LLOQ (1.00) 1.85 <LLOQ (1.00) <LLOQ (1.50) <LLOQ (1.00) 1.82 <LLOQ ( 1.00) 4.38 <LLOQ(1.00) 1.53 <LLOQ (1.00) 1.82 <LLOQ (1.00) <LLOQ (1.50) <LLOQ(1.00) 2.57 <LLOQ. (1.00) 5.54 <LLOQ (1.00) <LLOQ (1.50) <LLOQ (1.00) <LLOQ (1.50) <LLOQ (1.00) <LLOQ (1.50) <LLOQ(1.00) 1.58 <LLOQ ( 1.00) <LLOQ (1.50) <LLOQ (1.00) 2.29 <LLOQ (1.00) 2.10 <LLOQ (1.00) 2.31 <LLOQ ( 1.00) <LLOQ (1.50) PFOA* 3.60 6.01 3.59 3.21 6.16 6.97 4.44 7.22 4.74 3.15 6.41 7.20 4.21 3.69 6.65 5.40 3.01 7.23 10.5 4.16 4.75 5.95 7.78 4.62 8.95 3.72 5.38 3.96 2.21 7.45 6.90 3.52 7.81 8.07 3.89 2.57 PFHS* M556* M570* 2.67 <LLOQ (2.50) 1.67 3.66 <LLOQ (2.50) 2.38 5.79 <LLOQ (2.50) 1.09 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) 2.38 <LLOQ (2.50) 1.84 3.19 <LLOQ (2.50) 1.52 6.50 <LLOQ (2.50) 1.99 1.69 <LLOQ (2.50) 1.54 3.64 <LLOQ (2.50) 1.70 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) 2.06 <LLOQ (2.50) 1.43 <LLOQ (1.36) 4.09 1.82 3.72 <LLOQ (2.50) 1.08 1.95 <LLOQ (2.50) <LLOQ (1.00) 3.21 <LLOQ (2.50) 3.39 3.78 <LLOQ (2.50) 1.43 <LLOQ (1.36) <LLOQ (2.50) 1.41 <LLOQ (1.36) <LLOQ (2.50) 3.44 5.42 <LLOQ (2.50) 1.50 3.12 <LLOQ (2.50) 1.67 2.47 <LLOQ (2.50) 1.53 2.62 <LLOQ (2.50) 2.23 3.15 <LLOQ (2.50) 1.48 5.50 <LLOQ (2.50) <LLOQ (1.00) 2.27 <LLOQ (2.50) 1.44 2.77 <LLOQ (2.50) 1.45 10.6 <LLOQ (2.50) 1.78 <LLOQ (1.36) <LLOQ (2.50) 1.60 <LLOQ (1.36) <LLOQ (2.50) <LLOQ(1.00) 2.55 <LLOQ (2.50) 1.97 40.3 <LLOQ (2.50) 6.56 3.94 <LLOQ (2.50) 1.10 3.85 <LLOQ (2.50) 2.08 6.92 <LLOQ (2.50) 1.90 2.21 <LLOQ (2.50) 1.01 <LLOQ (1.36) <LLOQ (2.50) 1.49 NOTE: The method acceptance criteria states that the measured concentration for each QC must be within 20% (25% for PFOSA and PFOSAA) of the target concentration. The statistical error associated with any individual result may exceed the QC acceptance criteria. * Serum sample results for PFOA, PFHS, PFOSAA, PFOSA, M556 and M570 obtained using plasma calibration curves may vary from results obtained using serum calibration curves (see Results and Discussion section). Page 41 Northwest Bioanalytieal Study No. NW BS00-I45 Report No. NWBROl-155 Table 22. Serum Study Sample Concentrations (Continued) All concentrations are expressed as ppb. Sample Name 11509 11510 11511 11512 11513 11514 11515 11516 11517 11518 11519 11520 11521 11522 11523 11524 11525 11526 11527 11528 11529 11530 11531 11532 11533 11534 11535 11536 11537 11538 11539 11540 11541 11542 GH11281 GHl 1282 PFOS 22.9 46.7 161 54.4 14.5 20.4 26.5 37.6 49.2 8.50 19.5 25.3 18.6 35.2 28.7 20.1 23.9 49.8 15.4 31.1 15.6 19.3 21.2 40.4 25.5 20.5 23.2 38.6 15.4 33.4 32.9 15.7 24.4 40.9 36.2 18.2 PFOSA* PFOSAA* <LLOQ (1.00) 1.59 <LLOQ ( 1.00) <LLOQ (1.50) <LLOQ (1.00) 5.99 <LLOQ (1.00) <LLOQ (1.50) <LLOQ ( 1.00) 3.03 <LLOQ(1.00) 1.58 <LLOQ (1.00) <LLOQ (1.50) <LLOQ ( 1.00) <LLOQ (1.50) <LLOQ ( 1.00) <LLOQ (1.50) <LLOQ (1.00) <LLOQ (1.50) <LLOQ(1.00) <LLOQ (1.50) <LLOQ (1.00) 1.92 <LLOQ(1.00) 1.83 <LLOQ (1.00) <LLOQ (1.50) <LLOQ (1.00) 1.80 <LLOQ (1.00) <LLOQ (1.50) <LLOQ(1.00) <LLOQ (1.50) cLLOQ(l.OO) 2.30 <LLOQ (1.00) <LLOQ (1.50) <LLOQ ( 1.00) <LLOQ (1.50) <LLOQ (1.00) <LLOQ (1.50) <LLOQ (1.00) <LLOQ (1.50) <LLOQ ( 1.00) <LLOQ (1.50) cLLOQ(l.OO) 1.62 <LLOQ ( 1.00) <LLOQ (1.50) cLLOQ(l.OO) <LLOQ (1.50) <LLOQ (1.00) <LLOQ (1.50) <LLOQ (1.00) 1.86 <LLOQ (1.00) <LLOQ (1.50) <LLOQ ( 1.00) <LLOQ (1.50) <LLOQ ( 1.00) <LLOQ (1.50) cLLOQ(I.OO) <LLOQ (1.50) <LLOQ ( 1.00) <LLOQ (1.50) <LLOQ ( 1.00) <LLOQ (1.50) <LLOQ (1.00) <LLOQ (1.50) <LLOQ ( 1.00) 1.68 PFOA* 5.53 6.62 14.2 5.07 4.58 2.12 2.97 5.83 9.56 1.57 2.10 3.63 2.48 5.36 4.73 3.48 3.61 5.51 2.82 4.43 2.15 3.18 2.99 4.76 1.88 2.20 2.62 6.95 2.43 2.38 4.40 2.25 3.69 3.77 5.91 2.58 PFHS* M556* M570* 2.23 <LLOQ (2.50) <LLOQ (1.00) 5.35 <LLOQ (2.50) 1.13 4.61 <LLOQ (2.50) <LLOQ (1.00) 3.30 <LLOQ (2.50) 2.87 8.03 <LLOQ (2.50) 1.20 <LLOQ (1.36) <LLOQ (2.50) 2.86 4.86 <LLOQ (2.50) 1.54 4.35 <LLOQ (2.50) 1.05 2.03 <LLOQ (2.50) 2.48 3.72 <LLOQ (2.50) <LLOQ (1.00) 2.35 <LLQQ (2.50) 1.53 <LLOQ (1.36) <LLOQ (2.50) 2.70 1.71 <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.36) <LLOQ (2.50) cLLOQ(l.OO) 2.39 <LLOQ (2.50) 1.21 1.37 <LLOQ (2.50) <LLOQ (1.00) 1.38 <LLOQ (2.50) <LLOQ (1.00) 6.41 <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) 3.99 <LLOQ (2.50) 2.05 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) 2.71 <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.36) <LLOQ (2.50) 2.46 2.16 <LLOQ (2.50) 2.57 1.46 <LLOQ (2.50) <LLOQ (1.00) 1.38 <LLOQ (2.50) <LLOQ (1.00) 1.49 <LLOQ (2.50) <LLOQ (1.00) 9.60 <LLOQ (2.50) 1.26 <LLOQ (1.36) <LLOQ (2.50) 1.45 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) 2.28 <LLOQ (2.50) <LLOQ ( 1.00) <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) 21.8 <LLOQ (2.50) 3.10 2.68 <LLOQ (2.50) 2.00 3.10 <LLOQ (2.50) 2.13 <LLOQ (1.36) <LLOQ (2.50) 2.49 NOTE: The method acceptance criteria states that the measured concentration for each QC must be within 20% (25% for PFOSA and PFOSAA) of the target concentration. The statistical error associated with any individual result may exceed the QC acceptance criteria. * Semm sample results for PFOA, PFHS, PFOSAA, PFOSA, M556 and M570 obtained using plasma calibration curves may vary from results obtained using serum calibration curves (see Results and Discussion section). Page 42 Northwest Bioanalytical Study No. NWBS00-145 Report No. NWBR01-155 Table 22. Scrum Study Sample Concentrations (Continued) All concentrations are expressed as ppb. Sample Name GH11283 GH11284 GH11285 GH11286 GH11287 GH11288 GH11289 GH11290 GH11291 GH 11292 GH11293 GH 11294 GH 11295 GH11296 GH11297 GH11298 GH 11299 GH 11300 GH11301 GH11302 GH 11303 GH11304 GH11305 GH 11306 GH 11307 GH 11308 GH 11309 GH11310 GH11311 GH11312 GH11313 GH11314 GH11315 GH11316 GH 11317 GH11318 PFOS 13.8 18.7 21.1 29.4 52.0 40.5 37.7 22.7 41.3 62.6 9.58 20.3 16.3 30.0 21.8 54.3 15.4 27.2 27.1 15.7 30.5 16.2 11.7 21.2 36.1 35.7 21.2 44.8 103 13.4 27.4 26.0 31.8 27.1 39.7 37.2 PFOSA* PFOSAA* PFOA* PFHS* M556* M570* <LLOQ ( 1.00) <LLOQ (1.50) 2.29 3.37 <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.00) <LLOQ (1.50) 3.60 5.20 <LLOQ (2.50) 1.13 <LLOQ ( 1.00) <LLOQ (1.50) 3.82 1.96 <LLOQ (2.50) 2.52 <LLOQ (1.00) 3.09 3.69 1.83 <LLOQ (2.50) cLLOQ(l.OO) <LLOQ (1.00) 1.74 4.55 <LLOQ (1.36) <LLOQ (2.50) 4.24 <LLOQ ( l .00) <LLOQ (1.50) 7.41 1.78 <LLOQ (2.50) 1.78 <LLOQ (1.00) 1.81 6.69 2.94 <LLOQ (2.50) 2.62 <LLOQ ( 1.00) 3.25 4.13 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.00) 1.54 4.53 8.76 <LLOQ (2.50) 1.04 <LLOQ (1.00) 2.09 6.56 5.25 <LLOQ (2.50) 3.48 <LLOQ (1.00) 5.06 2.04 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) <LLOQ(1.00) <LLOQ (1.50) 5.59 1.70 <LLOQ (2.50) 1.61 <LLOQ (1.00) <LLOQ (1.50) <LLOQ ( 1.44) 7.23 <LLOQ (2.50) 2.82 <LLOQ (1.00) 4.29 5.24 3.08 <LLOQ (2.50) 1.91 <LLOQ (1.00) 1.53 3.96 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) <LLOQ(1.00) 1.68 6.52 2.84 <LLOQ (2.50) 4.11 <LLOQ ( 1.00) <LLOQ (1.50) 2.55 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.00) 1.77 4.28 1.85 <LLOQ (2.50) 1.02 <LLOQ ( 1.00) 2.61 6.88 3.18 <LLOQ (2.50) 2.09 <LLOQ ( 1.00) <LLOQ (1.50) 3.17 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.00) 1.61 3.73 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.00) 3.75 1.79 2.03 <LLOQ (2.50)"1 1.49 <LLOQ ( 1.00) 2.24 3.44 <LLOQ (1.36) <LLOQ (2.50) 1.43 <LLOQ ( 1.00) <LLOQ (1.50) 3.22 <LLOQ (1.36) <LLOQ (2.50) 2.37 <LLOQ ( 1.00) <LLOQ (1.50) 3.92 <LLOQ (1.36) <LLOQ (2.50) 1.40 <LLOQ (1.00) 2.05 . 3.34 1.90 <LLOQ (2.50) 2.52 <LLOQ (1.00) <LLOQ (1.50) 3.04 10.6 <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.00) 1.64 3.50 2.01 <LLOQ (2.50) 1.77 <LLOQ (1.00) 7.65 15.0 6.84 <LLOQ (2.50) 2.79 <LLOQ (1.00) <LLOQ (1.50) 3.47 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) <LLOQ ( 1.00) <LLOQ (1.50) 4.08 2.74 <LLOQ (2.50) 2.12 <LLOQ ( 1.00) <LLOQ (1-50) 4.59 2.75 <LLOQ (2.50) 5.07 <LLOQ (1.00) <LLOQ (1-50) 3.81 2.11 <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.00) <LLOQ ( 1.50) 3.86 . 2.78 <LLOQ (2.50) 1.59 <LLOQ (1.00) 2.06 5.33 3.90 <LLOQ (2.50) 1.75 <LLOQ ( 1.00) 1.66 3.56 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) NOTE: The method acceptance criteria states that the measured concentration for each QC must be within 20% (25% for PFOSA and PFOSAA) of the target concentration. The statistical error associated with any individual result may exceed the QC acceptance criteria. * Serum sample results for PFOA, PFHS, PFOSAA, PFOSA, M556 and M570 obtained using plasma calibration curves may vary from results obtained using serum calibration curves (see Results and Discussion section). Page 43 Northwest Bioanalytical Study No. NWBSOO-145 Report No. NWBRO1-155 Sample Name GHi 1319 GH11320 GHI 1321 GHI 1322 GHI 1323 GHI 1324 GHI 1325 GHI 1326 GHI 1327 GHI 1328 GHI 1329 GHI 1330 Table 22. Serum Study Sample Concentrations (Continued) All concentrations are expressed as ppb. PFOS 34.1 21.6 43.5 104 10.9 27.6 17.7 45.6 69.6 36.4 12.8 27.3 PFOSA* PFOSAA* <LLOQ (1.00) <LLOQ (1.50) <LLOQ (1.00) <LLOQ (1.50) <LLOQ (1.00) 6.67 <LLOQ ( 1.00) 9.85 <LLOQ (1.00) 2.94 <LLOQ ( 1.00) 2.69 <LLOQ (1.00) <LLOQ (1.50) <LLOQ (1.00) 11.1 <LLOQ (1.00) 2.37 <LLOQ (1.00) <LLOQ (1.50) <LLOQ (1.00) <LLOQ (1.50) <LLOQ ( 1.00) <LLOQ (1.50) PFOA* 4.24 3.91 6.96 15.4 2.12 4.55 2.96 5.89 8.38 3.35 3.22 3.03 PFHS* M556* M570* 2.55 <LLOQ (2.50) 1.29 2.97 <LLOQ (2.50) <LLOQ (1.00) 2.19 <LLOQ (2.50) 1.04 3.73 2.75 1.06 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) 3.58 <LLOQ (2.50) 1.30 <LLOQ (1.36) <LLOQ (2.50) 1.16 6.02 <LLOQ (2.50) 2.06 2.39 <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) <LLOQ (1.36) <LLOQ (2.50) 1.53 1.92 <LLOQ (2.50) <LLOQ (1.00) ) NOTE: The method acceptance criteria states that the measured concentration for each QC must be within 20% (25% for PFOSA and PFOSAA) of the target concentration. The statistical error associated with any individual result may exceed the QC acceptance criteria. * Serum sample results for PFOA, PFHS, PFOSAA. PFOSA, M556 and M570 obtained using plasma calibration curves may vary from results obtained using serum calibration curves (see Results and Discussion section). Page 44 Northwest Bioanaiytical Study No. NWBSOO-145 Report No. NWBROl-155 Figure 1. Representative Calibration Curve for PFOS Analytical Run 6 analyzed on 15-Nov-200I Calibration Standards for PFOS (ppb) Regression Method = QUADRATIC - Weighting Factor = l/X**2 Quadratic Limit = 4300 Figure 2. Representative Calibration Curve for PFOA Analytical Run 6 analyzed on 15-Nov-2001 Calibration Standards for PFOA (ppb) Regression Method = QUADRATIC - Weighting Factor = 1/X**2 Quadratic Limit = -4270 Page 45 Northwest Bioanalytical Study No. NWBSOO-I45 Report No. NWBROl-155 Figure 3. Representative Calibration Curve for PFHS Analytical Run 6 analyzed on I5-Nov-200I Calibration Standards for PFHS (ppb) Regression Method = QUADRATIC - Weighting Factor = l/X**2 Quadratic Limit = 2200 Figure 4. Representative Calibration Curve for PFOSAA Analytical Run 6 analyzed on 15-Nov-200I Calibration Standards for PFOSAA (ppb) Regression Method * QUADRATIC - Weighting Factor = I/X**2 Quadratic Limit = -3640 Page 46 Northwest Bioanalytical Study No. N W B S 0 0 - 1 4 5 Report No. NWBRO1-155 Figure 5. Representative Calibration Curve for PFOSA Analytical Run 6 analyzed on IJ-Nov-2001 Calibration Standards for PFOSA (ppb) Regression Method = QUADRATIC - Weighting Factor = l/X**2 Quadratic Limit = -2180 0 50 100 150 200 250 300 350 400 450 500 Nominal Cone. Figure 6. Representative Calibration Curve for M556 Analytical Run 6 analyzed on 15-Nov-2001 Calibration Standards forM556 (ppb) . Regression Method = QUADRATIC - Weighting Factor = 1/X**2 Quadratic Limit = -2310 0 50 100 150 200 250 300 350 400 450 500 Nominal Cone. Page 47 Northwest Bioanalytical Study No. NWBS00-145 Report No. NWBRO1-155 Figure 7. Representative Calibration Curve for M570 Analytical Run 6 analyzed on 15-Nov-2001 Calibration Standards for M570 (ppb) Regression Method = QUADRATIC - Weighting Factor = l/X**2 Quadratic Limit = -12500 Instrument Response Nominal Cone. Page 48 Northwest Bioanalytical Study No. NWBS00-I45 Report No. NWBR01-155 Figure 8. Human Plasma Blank for PFOS G ros I Internai Standard: H ffQ S Use Area Absolute Retention Time Expeaed RT S:06 Current Method Noise Threi 4.0 4.0 Quant Thre t o 1.0 Min. VMd S S Muk. Wid 20 20 Base. Widt 100 too RT Win. (sec 20 20 Smootl ! 1 S00I45OI037 1 017 S00143 CONTROL_0LANW4o Comment IT H ffO S I use is Internal Standard Expected RT 4:31 Current Method Noise Thre* _ 20.0 Quant Thre _ 4.C 4.0 _10Min. Wid ___ 5 MuH. Wid I 5 10 Base. Widt 100 RT Win. (se a , 25 100 2S Smootl S0014501037 1 037SOOI4S C O N TRO l_BlAN 4o Comment Page 49 Northwest Bioanalytieal Study No. NWBS00-145 Report No. NWBR01-155 Figure 9. Human Plasma Blank for PFOA IPfOA I Internal Standard: TMPf05 Use Area Absolute Retention Time Expected RT 4:37 Curren^ Method Noise Thiet 10.0 Quant Thie Min. Wid Muli. Wid Base. Widi 100 RT Win. (secs Smoot! 26 1 $0014501037 I 037 SOOI45 CONTROL BLANWdo Comment ITHPfOS I use as Internal Standard Expected RT 4:31 Current Method Noise Thre* 20.0 Quant Thre 4.C 4.0 Min. Wid ___ 5 3 _10 10Mult. Wid ' 100Base. Widt 100 RT Win. (see , Smootl 25 I S0014501037 1 037 $00143 CONTROL_BLANIQdo Comment Page 50 Northwest Bioanalytical Study No. NWBSOO-145 Report No. NWBROl-155 Figure 10. Human Plasma Blank for PFHS IPfMb 1 Internal Standard: THPFOS Use Area Absolute Retention Time Expected RT 3:56 Current Method Noise nirei 5. 5.0 Quant Threi 1.5 1.5 Min. WkJ 5 5 Mult. Wid IB 19 Base. Widt 100 100 RT Win. (tea 50 50 Smoot' 1 1 ITH P fO S u$c at Internal Standard Expeced RT 4:31 Current Method Noise ihre 20.0 Quant Thre Min. Wid 4.0 s Mult. Wid 10 Base. Wid! 100 RT Win. (sea 25 Smodi Page 51 Northwest Bioanalytical Study No. NWBSOO-145 Report No. NWBRO1-155 FigOure 11. Human Plasma Blank for PFOSAA ITHPfOS 1 ute at Internal Standard Expected RT O l Current Method Noise Threi 20.0 Quant Ihre 4.C 4.0 Min. Wid 5 5 Mult. Wid' 10 Base. Widt 100 100 RT Win. (tea 25 25 Smoot Page 52 Northwest Bioanalytical Study No. NW BS00-I45 Report No. NWBROI-155 FigOure 12. Human Plasma Blank for PFOSA W O SA I Internal Standard: THPfOS U m Area Absolute Retention Firne Expected RT 5:51 Current Method Noise Thre* Quart ihre Min. Wid Mult, vsr.d Base. Widt _ i; 100 7.0 1.5 S 17 100 RT Win. (sea _ 20 ISmooli ___ 20 1 It h t o s u*al.m *ISU n d<l I Current Method Noise Thre: 20.C 20.0 Q u TM . 4.0 4.0 Min. Wid 5 s Mult. Wid 10 to 100 too RF Win. (sea 25 25 Smotti 1 1 Page 53 Northwest Bioanalytical Study No. NWBSOO-145 Report No. NWBRO1-155 Figure 13. Human Plasma Blank for M556 | M 5 t ] Internal Standard; THPFOS Ute Area Absolute Retention Time Expected RT 5:36 Current Method Noise Titre 5.0 5.0 Quant Due 0.5 0.S Min. Wid 5 $ Mult. Wld 21 21 Bate. W.di too 100 RT Win. (tea 20 20 Smoot! 1 1 ITHPfOS tse as Internal Standard Expected RT 4:3! Current Method Notte Thre: 2o.a 20.0 Quant Thre 4.0 4.0 Min. Wid 5 5 Mult. Wld 10 to Bate. Wtdt 100 100 RT Win. (sea 25 25 Smoot 11 50014501037 1 037 S00145 CONTROL,BLANW4o Comment Fri. Nov 9, 2001 3:1 AM 8:59 in 1 period 100* 237 1: 8:59 M RM . 642 scant 427.0->407.0 Area 0 Heicht 0 Stan Time End Time Intetrarion Width Retention Time Intettration Type ItltMH 0:00.0 0:00.0 0:00.0 (hOO.O tfnsi(y:59eps v. 121 lai 241 301 361 1 41 541 SOI Scmjj031 1:42 232 331 4:13 501 304 6:44 7:35 9 :3 ;TM i Page 54 Northwest Bioanalytical Study No. NWBS00-145 Report No. NWBROI-155 Figure 14. Human Plasma Blank for M570 |M570 1 Internal Standard: fHPfOS Ute Area Absolute Retention Time Expected RT 5:54 Current Method Norse Hires SO s.o Quant Ihre \X 1.0 Min. VVid s 5 Muk. Wid 20 20 Base. Wtdi 100 100 RT Win. (sea 20 20 Smoot 1 1 Page 55 Northwest Bioanalytical Study No. NWBS00-145 Report No. NWBRO1-155 Figure 15. Human Plasma Blank with Internal Standard (QCO) for PFOS tos i Ute Aree Absolute Retention Time Expected RT 5:06 Current Method N o i Thres 4.U 4.0 Quam Ihre Min. Will Mult. WkJ Base. WkJt RT Win. tsea 5s 20 20 100 100 20 20 Smodi 1 1 ITHPfOS use at Internai Standard Expeaed RT 4:31 . Current Method Noise Threi 20.0 20.0 Quant Threi 4.0 4.0 Min. Wid S 5 Mult Wid 10 10 Base. Widt 100 100 RT Win. (sec 25 25 Smooti 1 1 S0 0M 50101 1 o n SOOHS Q C O INo Comment Page 56 Northwest Bioanalytical Study No. NWBS00-I45 Report No. NWBRO1-155 Figure 16. Human Plasma Blank with Internal Standard (QCO) for PFOA [S 1 Internal Standard: THPfOS Use Area Absolute Retention Time Expected RT 4:37 Current Method Notse Thre 10.( 10.0 Quant Thre 3.0 3.0 Min. Wid 8 8 Mult. Wid 16 16 Base. WkJi 100 100 RT Win. (sea 26 26 Smooll 1 1 ) iNo Comment 1T H P f O S 1 use as Internai Standard Current Method Noise Thres 20.0 20.0 Q_uan_t T__hre 4.0 4.0 Min. Wid 5 5 Muit. Wid 10 10 Expeoed RT 4:31 " - widl 100 100 RT Win. (sea 2S 25 Smodi 1 1 $0014501011 1 O il $00145 Q C INo Comment Page 57 Northwest Bioanalyticai Study No. NWBSOO-145 Report No. NWBRO1-155 Figure 17. Human Plasma Blank with Internal Standard (QCO) for PFHS F hs I internal Standard: THWOS Use Area Absolute Retention Time Expeced RT 3:56 Current Method Noise Threi 5.0 5.0 Quant Thre I.S 1.5 Min. Wid 5 5 Muit. Wid IB 18 Base. Widi too 100 RT Win. (sea 50 SO Smootl t 1 SOOM5010U 1 O ll S00145 Q C O INO Comment jTHPfQS us as Internai Standard Expected RT 4:31 Current Method Noise Thre: _ 20.0 Quant Thre _ 4.0 Min. Wid 5 Muit. Widi 10 10 >00Base. Wkjt 100 RT Win. (sec , 25 25 Smootl $0014501011 1 O il SOOI4S Q C O INoComment Page 58 Northwest Bioanalytieal Study No. NWBSOO-145 Report No. NWBRO1-155 Figure 18. Human Plasma Blank with Internal Standard (QCO) for PFOSAA (PfOSAA 1 Internal Standard: THPFOS Use Area Absolute Retention Time Eapeoed RT 6:03 Current Method Notte ThreS 5.0 5.0 Quant Thre 1.3 1.3 Min. Wid 6 6 Mult. Wid 17 17 Bate. Widt 100 100 RT Win. (sea 20 20 Smooil 1 1 SO0M5O1011 I 01 1 SOOU5 Q C O INo Continent Thu. Nov 8. 2001 10:17 PM 8:59 in 1 period 1: 8:59 MRM, 642 scans SS4.0>4I9.0 Area 4592 Heifhf 590 Stan Time End Time Integration Width Retention Time Integration Type 5:53 6:18 0:25.2 6:07 A-BB 8. ITHPFOS use as Internal Standard Expeaed RT 4:31 Current Noise Thre: 20.C Quant Thre 4.0 Min. Wid 5 Mult. Wid 10 Base. Widt 100 RT Win. (sea Smootl 25 1 20.0 4.0 5 10 100 25 1 SCOI 4501011 1 011 $001*5 QCO IN Comment Page 59 Northwest Bioanalytical Study No. NWBSOO-145 Report No. NWBRO1-155 Figure 19. Human Plasma Blank with Internal Standard (QCO) for PFOSA 1PFOSA 1 Internal Standard: THPfOS Use Area Absolute Retention time Expected R t S:S 1 Current Method No m Thre 7.0 7.0 Quant Thre 1.5 1.5 Min. Wid 5 5 Mull. Wid J7 7 Bate. Widt too too RT W n, (secs 20 20 Smooti 1 1 $0014501011 1 OM $00145 Q C O iNo Comment IOO........Thu. Nov S. 2001 10:17 PM 8:59 in t period I: 8:59 M RM . 642 scam 497.9*>77.9 Area 0 Heieht 0 Start Time End Time Intecration Width Retention Time Intetration Type 0:000 0:00.0 0:00.0 0:00.0 fituttrty: 174cpj ITHPFOS u m as Internal Standard I Current Method Noise Thre: 200 20.0 Quant Thre 4.C 4.0 Min. Wid 5 5 Mult. Wid 10 10 Expected RT 4:31 Base. Widt too too RT Win. (sea 25 25 Smoot: 1 t S0014SOI011 I 01t SOOI4S Q CO IN o Comment 4S;r8441;::'-T6305:11:i8.5131 Page 60 Northwest Bioanalytieal Study No. NWBS00-I45 Report No. NWBRO1-155 Figure 20. Human Plasma Blank with Internal Standard (QCO) for M556 |MS56 1 Ute Area Absolute Retention Time Current Method Notte Ihre $.0 5.0 Quant Thre 0.5 Min. Wid s S Mult. Wid 21 2t Base. Widt 10O 100 KT Win. (seo 20 20 Smorti 1 1 SO0M 5O IOM I Ot 1 $00145 Q C O IN o Comment ITHPfOS use as Internal Standard Expeaed RT 4:31 Current Method Noise Thre 20.0 20.0 Quant Thre 4.0 4.0 Min. Wid S 5 Mult. Wid TO IO Base. Widt 100 100 RT Win. (sea 25 25 Smorti 1 1 $0014501011 1 Ot 1 S00145 Q C O IN o Comment ' Page 61 Northwest Bioanalytical Study No. NWBS00-I45 Report No. NWBROl-155 Figure 21. Human Plasma Blank with Internal Standard (QCO) for M570 MS70 | Interne! Standard: THPFOS Use Area Absolute Retention Time Expeed RT 5:54 Current Method Noise Thres 5.0 5.0 Quant Bite i.r 1.0 Min. Widl 5 5 Mult. Wid 20 20 Base. Widl 100 100 RT Win. (sec 20 20 Smoot; 1 1 $0014501011 1 O U SOOMS Q C O W o Comment Thu. Nov 8, 2001 10:17 PM 8:59 in 1 period 1: 8:59 M RM . 642 sa n s S70.0>>419.0 Area 1050 Heieht 217 Start Time End Time Intetration Width Retention Time Integration Tvoe 5:47 5:60 0:12.6 5:54 A-BB htmsity: 233 ep ; ' . 6 18t : -241 501;" 61 --42?;- :',48i;. ;54?.:..:-:60|;.: F-42:!j-2;S2-;;;;23:-H 4;I3 3f l :-J:54-.^;44::;i-7:33 <::$:23[ ITMPfOS use as Internal Standard Expeaed RT 4:31 Current Method Noise Thre- 20.C 20.0 Quant Thre 4.0 4.0 Mm. Wid 5 5 Mult. Wid 10 10 Base. Widt 100 100 RT Win. (sec 25 25 Smoot 11 S001450IOH 1 011 500143 Q C O INo Comment Page 62 Northwest Bioanalytieal Figure 22. Low Standard for PFOS Ipfs 1 Internet Standerd: THPfOS Ute Area Absolute Retention Time Expeaed RT 5:06 Current Method Notte Ihres 4.0 4.0 Quatti Thre Min. Wtd 5 5 Mult. Wid 20 20 Bate. Wkit 100 too RT Win. (tea 20 20 Smoot 1 1 t S0W 50I001 1 001 S00I4S Stdl 1 1 No Comment Study No. NWBSOO-145 Report No. NWBROi-155 It h p f o s _ Expeaed RT e j l I Current Method Notte Ihres 20.0 20.0 Quant Thre 4.C 4.0 Min. Wid s 5 Muk. Wid 10 10 B *e . Wid' too 100 RT Win. (secs 25 25 Smootl 1 1 S00US01001 1 001 SOOI45 Stdl 1 I No Comment Page 63 Northwest Bioanalytical Figure 23. Low Standard for PFOA Study No. NWBS00-145 Report No. NWBRO1-155 IPfOA 1 Internal Standard: HPfOS Ute Area Absolute Retention Time Expected RT 4:37 Cunen* Method Noise Thre Quant Ihre Min. Wid 1U.0 3.0 8 Mull. Wid 16 Base. W kJi RT Win. (sea 100 36 Smodi Thu, Nov 8. 3001 8:25 PM 8:59 in I period 1: 8:59 M RM . 642 scans 413.0- >169.0 Area 9405 Heieht 1095 Stan Time End Time Inteeration Width Retention Time Inteeration Type 4:19 4:48 0:29.4 4:37 A BB ntMSity;IH4cpf IT H P fO S I use as Internal Standard Expected RT 4:31 Current Method Noise Thre 20.0 20.0 Quant Thre 4.0 4.0 Min. Wid 5 S Mult. Wid 10 10 Base. Widi 100 100 RT Win. (sea 25 25 Smoot 11 Page 64 Northwest Bioanalytical Figure 24. Low Standard for PFHS IWMS I Internai Standard: TMPFOS Ute Area Absolute Retention Time Expected RT 3:56 Current Method Notte Ihre Quant Thre 5.0 1.5 Min. WkJ Muli. Wid 150 Baie. Widt 100 RT Win. (tea _ Smootl SO 1 $0014501001 I 001 S00I45 Std l t 1 No Comment Study No. NWBS00-145 Report No. NWBR01-155 ITHPfOS I Current Method ) use as Internal Standard Noise Hire 20.0 20.0 Quant Hire 4.0 4.0 Min. WkJ 5 5 Mult. Wid 10 10 ExpeCed RT 4:31 Base. Widt RT Win. (sea 100 25 100 25 Smootl 1 1 S0014501001 1 00) $00145 S*d1 1 1 No Comment ) Page 65 Northwest Bioanalytical Figure 25. Low Standard for PFOSAA Study No. NWBS00-145 Report No. NWBR01-155 iPfOSAA 1 Internai Standard; THPF05 Use Area Absolute Retention Time Expected RT 6:03 Current Method Norse ihre 5.C 5.0 Quant Threi 1.3 1.3 Min. WkJ 6 6 Muil. Wid 1? 17 Base. Widt 100 100 RT Win. (secs 20 20 Smootl 1 1 $0014501001 1 001 S00I4S S(d1 1 1 No Comment Thu, N ov S. 2001 8:25 PM 8:59 in 1 period 100 1: 8:59 M RM . 642 scans 90 SB4.O*>419.0 ; 90 Area 6942 Heieht 943 Start Time End Time Intetration Width Retention Time Intetration Type 70- 5:47 6:15 0:27.7 6:06 A BB ; sijrr|i';ypi ! ,,40- : 4 35.: htffi*ity:968cpi _!? ;;2b10- 23 5_3 154 208 243 279. ' 351 , 97, V ta t: M8- il- 241 :;301.: 361 - 42t "481::- 54 y ... :Q\ 051 j a ; S:44':V.T35 ,,;$;25: [ ITHPFOS use as internai Standard Expected RT 4:31 Current M ethod Noise Ihr 20.0 Quant Thre 4.0 Min. Wid 5 Mult. Wid 10 Base. Wich 100 RT Win. (sea 25 Smootl $0014501001 1 001 $00145 Stdl 1 1 No Comment Page 66 Northwest Bioanalytical Fi9gure 26. Low Standard for PFOSA Study No. NWBSO-145 Report No. NWBR01-I55 Ip f o s a I Internai Standard: fHPFOS Use Area Absolute Retention Time Expeaed RT 5:51 Current Method Noise Thre 7.0 7 0 Quant Thre 1.5 1.5 M ia Wid 5 S Mult. Wid 17 17 Base. Widt 100 100 RT Win, (seo 20 20 Smodi 1 1 $0014501001 t 001 SOOJ45Sidl 1 1 No C Thu. Nov 8. 2001 8:25 PM 8:59 in 1 period 100 1: 8:59 MRM . 642 scant ; 30 497.9*> 77.9 90 Area 4999 Heitht 55 7 Start Time End Time Integration Width Retention Time Intetration Type 5:33 5:57 0:24.4 5:52 A-BB 70' 60 50 40- intentrty; 583 eps 419 :3b- "2b* Tp- 38 7 , 128 , 200 .4 5i 3 ..3. 04: ^560 :- $cZ'3n-. Xfi - 6 : 12! 181 : 24V 31: !:J6I;i: :42t v::;48f - -:54r- $01 Se*w- 051-i-vi;42 ""2:32:r' aSXi i -MJ-; 8 ;44-i,T im r iTHPfOS use at Internal Standard Expeaed RT 4:31 Current Method Noise Thre: Quant Thre 20.0 4 .0 20.0 4.0 M ia Wid 5 s Mult. Wid 10 10 Base. Writ 100 t o o RT Win. (sect 25 25 >1 $0014501001 1 001 $00145 Stdl 11 No Coi Thu. Nov 8. 2001 8:25 PM 8:59 in 1 period 1: 8:59 M RM . 642 scant 427.0->40 7.0 Area 836307 Bei(ht 117889 Start Time End Time lnteration Width Retention Time intetration Type 4:24 4:53 0:29.4 4:35 A-BB -" V - ;ntraity: 117938 eps 327. V-'i :;ri":;-3:!'; :SrK ..?..g " L;_ :!. - 36 86 12$ $1 121 0:31 1:42 187 252 278 181 241 301 361 2 3 2 3 2 3 4:13 5:05 428 501 542 421 431 541 3.54 $:44 7:33 $04 $01 S c w 9 5 Thwf Page 67 Northwest Bioanalytieal Figure 27. Low Standard for M556 |M556 1 Internal Standard: THPfOS Ute Area Absolute Retention Time ExpeOed RT 5:36 Current Method Notte Thre* S.O Quant Thre 0.5 Min. Wkf S Mull. Wid Base. Wk)i 21 100 RT Win. b e o _ Smooti 20 1 $0014501002 1 002 $00145 Sid2 1 1 No Comment Study No. NWBS00-145 Report No. NWBRO1-155 ITHPfOS I ute at Internal Standard Expected RT 4:31 Current Method Noise Ihres 20.0 20.0 Quant Thre 4.0 4.0 Min. Wid S s Mull. Wid 10 10 Base. Wid! 100 100 RT Win. (tea 25 25 Smooti 1 1 S001450I002 I 002 S00145 Std2 1 I No Comment Page 68 Northwest Bioanalytical Figure 28. Low Standard for M570 |M570 1 Internal Standard: fHPfOS Use Area Absolute Retention Time Expected RT 5:54 Curreiti Method Noise Thre 5.1 5.0 Quant Thre I.C 1.0 Min. Will 5 5 Mull. Wid 20 20 Base. Wldt too 100 RT Win. (secs 20 20 Smooti 1 1 50014501001 I 001 SOOI45Sld1 I 1 N o Comment Study No. NWBSOO-145 Report No. NWBRO1-155 iTHPfQS use at Internal Standard Expected RT 4:31 Current Method Noise Thret 20.0 20.0 Quant Threi 4.0 4.0 Min. Wid 5 5 Mull. Wid 10 10 Base. Widt too too RT Win. (sea 25 25 Smoot! 1 1 S001450100 1 001 SOOMS Stdl I I No Comment Page 69 Northwest Bioanalytical Figure 29. High Standard for PFOS IH-OS 1 Internal Standard: fHPFOS Use Area Absolute Retention Time Expeaed RT 5:06 Current Method Noise Thre*i 4.0 4.0 Quant ihre 1.0 Min. Wid S 5 Mull. Wid 20 20 Base. Widt 100 100 RT Win. (sea 20 20 Smootl 1 ! $0014501009 1 009 SOOUS Sld9 1 1 No Comment Study No. NWBSOO-145 Report No. NWBRO1-155 ITHPfQS use as Internal Standard Expeaed RT 4:31 Current Method Noise Thrc! 20.0 Quant Thre 4.0 Min. Wid Mult. Widi 150 Base. Widi 100 RT Win. (sea _ 25 Smootl S0014501009 1 009 S00145 Std9 I 1 No Comment Page 70 Northwest Bioanalytieal Figure 30. High Standard for PFOA (Pf 1 Internat Standard: TMPfOS Ute Area Current Method to.oNoise ihres 10.0 Quant Ihre 3.0 3.0 Min. WkJ S 8 Absolute Retention Time ExpeCed RT 4:J7 Mult. Wld 16 16 Base. Widt too too RT Win. (sea 26 26 Smootl 1 1 $00(4501009 I 0O9SOOI45 5ld9 I I No Comme Study No. NW BS00-I45 Report No. NWBR01-155 ITHPfOS use es Internai Standard Expected RT 4:31 Current Method Noise Threi 20.0 20.0 Quant Threi 4.0 4.0 Min. Wid 5 5 Mull. W mJ 10 IO Base. With too too RT Win. (sea 25 25 Smootl 1 1 SOO14501009 1 009 S00I45 Sld9 I 1 No Comment Page 7 I Northwest Bioanalytical Figure 31. High Standard for PFHS Internal Standard: THPFOS Ute Area Abtolute Retention Time Expected RT 3:56 G m e n i Method Noite Thret 5 0 5.0 Quant Dire 1.5 I.S Mm. Wid S 5 Muli. Wid ta 16 Bate. Wldt 100 100 RT Win. (ica 50 50 Smoot 11 50014501009 1 009 $00145 $k J9 11 No Comment Study No. NWBSOO-145 Report No. NWBR01-155 iTHPfOS Expeaed RT . 4:31 1 Current Method Notte Threi 20.0 20.0 Quant Thie 4.0 Min. Wid 5 S Mult. Wid 10 10 >* " dl RT Win. (sec Smootl to o 25 1 100 25 1 $0014501009 1 009 SOOI45 Std9 1 I No Comment ) Page 72 Northwest Bioanalytieal Figure 32. High Standard for PFOSAA Study No. NWBS00-145 Report No. NWBRO1-155 IPfOSAA 1 Internal Standard: THPFOS Ute Area Absolute Retention Time Current Method Nmse Thre 5.0 S.O Quant Thre 1J 1.3 Min. Wid 6 6 Mult. Wid 17 17 Base. Widt 100 100 RT Win. (sea 20 20 Smooi 1 1 S0014501009 1 009 $00145 Std9 1 1 No Comment ITHPfOS Expeaed RT 4:31 1 Current Method Noise Thre 20.0 20.0 4.0 Min, Wid 5 5 Mult. Wid 10 10 >* W 100 100 RT Win. (sea 25 25 Smootl 1 1 S0 0 14501009 1 009S00145 Std9 1 I No Comment Page 73 Northwest Bioanalytical Figure 33. High Standard for PFOSA Study No. NWBSOO-145 Report No. NWBRO1-155 IPfOSA 1 Internai Standard: H-tPfOS Use Area Absolute Retention Time Expected RT 5:51 Current Method Noise ihres 7.0 7.0 Quant Ihre 1.3 1.5 Min. Wid 5 5 Mull. Wid 17 17 Base. Widt 100 too RT Win. (sea 20 20 Smootl 1 S0OM510O9 1 009S00145$id9 I 1 No Comment ITHPfOS I me as Internai Standard Expected RT 4:3t Noise Thret Quant Ihre Min. Wkf Mult. Wid Base. Widt RT Win. (se a , Smootl Method 20.0 4.0 5 10 100 2S 1 S0014501009 1 009 $00145 Sid* 1 1 No Comme Page 74 Northwest Bioanalytical Figure 34. High Standard for M556 |MSS6 ~| Internal Standard: THPFOS U m Area Abtoiute Retention Time Exoeaed RT 5:3h Current Method Notte Threi 5.0 Quant fhre Min. Wkl Mult. Wid a Bate. Widt t 21 100 0.5 5 21 100 RT Win. (iea a 20 Smootl 20 1 50014501009 1 009 500145 Sld9 I t No Comment Study No. NWBS00-145 Report No. NWBROI-155 \ ITHPfOS I Current Method ; u m as Internal Standard N oiM Threj__ 20.0 20.0 Quant Thre 4,0 4.0 Min. Wid 5 Mult. Wid 10 Expeaed RT 4:31 Bate. Widt _ 100 100 RT Win. (te a , 25 25 Smootl S0014501009 1 009 S00145 Std9 1 1 No Comment Page 75 Northwest ioanalytical Figure 35. High Standard for M570 IMS 70 " Internal Standard: rupf OS Use Area Absolute Retention Time Expected RT 5:54 Current Method Noise Dire 5.0 5.0 Quant Hite* t.O 1.0 Min. Wid 5 5 Mult. Wid 20 20 Base. Wid! 100 t o o RT Win. (sea 20 20 Smootl 1 1 S0014501009 I 009 SOOMJ Sld I I No Comme Study No. NWBS00-145 Report No. NW BR0I-I55 ITHPFOS use as Internal Standard Expeaed RT 4:31 Current Method Noise Hire 20.0 Quant TTire 4.0 Min. Wid i 5 Mult. VMdi ' Base. Widt 100 RT Win. (sea _ Smootl S00I45O I0O 9 I 009 S0014S Sld9 1 1 N o Com m e Page 76