Document 1yNp0QEK7Dq1p9gXoBXzD3kpo

Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Quantitative Determination of PFOS and Related Compounds in Human Serum and Plasma by LC/MS/MS Sample Analysis Report for Protocol EPI-0012 Northwest Bioanalytical (NWB) A Division of NWT Inc. 1121 East 3900 South Salt Lake City, UT 84124 PREPARED FOR: 3M Company Epidemiology, Medical Department 220-3W-05 St. Paul, M N 55144 R u CZ> "o m ro "--. U4pOi C "J { rj --j'1" O vO O \0 AUTHOR: GdXsfLJW P __________________ DATE: Connie O. Sakashita, B.S., NWB Project Manager //-a o -Q / APPROVED FOR RELEASE BY: jl ^ ______________________________ D A T E : Rbuger L. Foltz, Ph.D., NWB Technical Director // - 2 1- 6 I Page 1 Northwest Bioanalytical QUALITY ASSURANCE STATEMENT Study No. NWBS00-091 Report No. NWBR01-065 LABORATORY: Northwest Bioanalytical (NWB) A Division o f NWT Ine. 1121 East 3900 South Salt Lake City, UT 84124 SPONSOR: 3M Company Epidemiology, Medical Department 220-3W-05 St. Paul, MN 55144 COMPOUND(S): PFOS and Related Compounds NWB STUDY NUMBER: NWBS00-091 SPONSOR STUDY NUMBER: EPI-0012 NWB STUDY TITLE: Quantitative Determination of PFOS and Related Compounds in Human Serum and Plasma 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 ot 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 Manager NWB Management* 16 Oct 2000 17-23 Oct 2000 29 Oct - 01 Nov 2001 23 Nov 2001 Analytical Plan Sample Analysis Report Draft and Raw Data Final Report 16 Oct 2000 24 Oct 2000 01 Nov 2001 23 Nov 2001 31 Oct 2000 31 Oct 2001 30 Nov 2001 30 Nov 2001 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. NWB Quality Assurance Compliance Auditor Date Page 2 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBR01-065 COM PLIANCE STATEM ENT The clinical study described in this report is not included w ithin 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 R egulations fo r N onclinical Laboratory Studies (Title 21 C FR 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 M HW Good Laboratory Practice Standard Ordinance for Nonclinical Laboratory Studies on the Safety o f Drugs (Ordinance No. 21, PAB Notification No. 424.) This report contains serum sample results. As reported in NW B report NW BR00-122 [5.3], serum sample results obtained using plasma curves for PFOSA, PFOSAA, POAA, PFHS, M 556 and M570 did not meet NWB SOP requirements for validation. Any known circumstances that may have affected the quality or integrity o f the project or reported data are discussed within the report. This report represents an accurate record o f the raw data. Connie O. Sakashita, B.S. NW B Project Manager _____21___________ huger L. Foltz, Ph.D. J NWB Technical Director //-= .2 Q - ! Date f ! - 2-t - o ) Date Page 3 Northwest Bioanalytical TABLE OF CONTENTS Study No. NWBS00-091 Report No. NWBRO1-065 SIGNATURE P A G E .........................................................................................................................................1 QUALITY A SSU R A N C E STA TEM EN T.................................................................................................. 2 COM PLIANCE S T A T E M E N T .....................................................................................................................3 TABLE OF C O N T E N T S ................................................................................................................................ 4 LIST OF T A B L E S ............................................................................................................................................4 LIST OF F IG U R E S ...........................................................................................................................................6 1. IN TR O D U C TIO N ....................................................................................................................................8 2. M E T H O D O L O G Y ...................................................................................................................................9 3. SAM PLE A N A LY SIS...........................................................................................................................12 4. RESULTS AN D D ISC U SSIO N ..........................................................................................................13 5. R E FE R E N C E S........................................................................................................................................16 6. D A TA R E T E N T IO N ............................................................................................................................16 LIST OF TABLES Table 1. Calibration Curve Sum m ary for P F O S ....................................................................................17 Table 2. Calibration Curve Sum m ary for PFO A .................................................................................... 18 Table 3. Calibration Curve Sum m ary for P F H S ....................................................................................19 Table 4. Calibration Curve Sum m ary for PFO SA A ............................................................................. 20 Table 5. Calibration Curve Sum m ary for PFOS A ............................................................................... 21 Table 6. Calibration Curve Sum m ary for M 5 5 6 .....................................................................................22 Table 7. Calibration Curve Sum m ary for M 5 7 0 .....................................................................................23 Table 8. Back-Calculated Concentrations o f Calibration Standards for P F O S ................................ 24 Page 4 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Table 9. Back-Calculated Concentrations o f Calibration Standards for P F O A ..............................25 Table 10. Back-Calculated Concentrations o f Calibration Standards for P F H S ..............................26 Table 11. Back-Calculated Concentrations o f Calibration Standards for P F O S A A .......................27 Table 12. Back-Calculated Concentrations o f Calibration Standards for P F O S A .......................... 28 Table 13. Back-Calculated Concentrations o f Calibration Standards for M 556..............................29 Table 14. Back-Calculated Concentrations o f Calibration Standards for M 570..............................31 Table 15. Analytical QC Summary for P F O S ......................................................................................... 33 Table 16. Analytical Q C Sum mary for P F O A ........................................................................................ 34 Table 17. Analytical Q C Sum mary for P F H S ..........................................................................................35 Table 18. Analytical QC Sum mary for P F O S A A ...................................................................................36 Table 19. Analytical QC Sum mary for P F O S A ...................................................................................... 37 Table 20. Analytical QC Sum mary for M 5 5 6 ..........................................................................................38 Table 21. Analytical QC Sum mary for M 5 7 0 ..........................................................................................39 Table 22. Serum Study Sam ple C oncentrations..................................................................................... 40 Table 23. Plasm a Study Sample Concentrations....................................................................................48 Table 24. R epeat Analysis Table for PFOS in Hum an S e ru m ........................................................... 55 Table 25. R epeat A nalysis Table for PFO A in H um an Serum ........................................................... 55 Table 26. R epeat Analysis Table for PFHS in H um an S e ru m ........................................................... 56 Table 27. R epeat Analysis Table for PFO SA A in H um an S e ru m ...................................................... 56 Table 28. R epeat Analysis Table for PFO SA in Hum an S e ru m ..........................................................57 Table 29. R epeat Analysis Table for M 556 in Hum an S e ru m ........................................................... 57 Table 30. R epeat A nalysis Table for M 570 in Hum an S e ru m ........................................................... 58 Page 5 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 LIST OF FIGURES Figure 1. Representative Calibration Curve for P F O S .........................................................................59 Figure 2. Representative Calibration Curve for P F O A ........................................................................59 Figure 3. Representative Calibration Curve for P F H S .........................................................................60 Figure 4. Representative Calibration Curve for P F O S A A .................................................................. 60 Figure 5. Representative Calibration Curve for P F O S A ..................................................................... 61 Figure 6. Representative Calibration Curve for M 5 5 6 .........................................................................61 Figure 7. Representative Calibration Curve for M 5 7 0 ............................................ 62 Figure 8. Hum an P lasm a Blank for P F O S .............................................................................................. 63 Figure 9. Hum an Plasm a B lank for P F O A .............................................................................................. 64 Figure 10. Hum an Plasm a B lank for P F H S .............................................................................................65 Figure 11. Hum an Plasm a Blank for PFO SA A ....................................................................................... 66 Figure 12. Hum an Plasm a Blank for PFO SA ...........................................................................................67 Figure 13. Hum an Plasm a Blank for M 5 5 6 .............................................................................................68 Figure 14. Hum an Plasm a Blank for M 5 7 0 .............................................................................................69 Figure 15. Hum an P lasm a Blank w ith Internal Standard (QCO) for P F O S ..................................... 70 Figure 16. Hum an Plasm a Blank w ith Internal Standard (QCO) for P F O A .................................... 71 Figure 17. Hum an Plasm a Blank w ith Internal Standard (QCO) for PFH S..................................... 72 Figure 18. Human Plasm a Blank w ith Internal Standard (QCO) for P F O S A A ...............................73 Figure 19. Hum an Plasm a Blank w ith Internal Standard (QCO) for PFO SA .................................. 74 Figure 20. Hum an Plasm a Blank w ith Internal Standard (QCO) for M 556..................................... 75 Figure 21. Hum an Plasm a Blank w ith Internal Standard (QCO) for M 570..................................... 76 Figure 22. Low Standard for P F O S ............................................................................................................77 Figure 23. Low Standard for P F O A .......................................................................................................... 78 Page 6 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Figure 24. Low Standard for P F H S ...........................................................................................................79 Figure 25. Low Standard for P F O S A A ....................................................................................................80 Figure 26. Low Standard for P F O S A .........................................................................................................81 Figure 27. Low Standard for M 55 6 ............................................................................................................ 82 Figure 28. Low Standard for M 5 7 0 ............................................................................................................ 83 Figure 29. High Standard for P F O S ........................................................................................................... 84 Figure 30. High Standard for P F O A ..........................................................................................................85 Figure 31. High Standard for P F H S ................................................................. 86 Figure 32. High Standard for P F O S A A .................................................................................................... 87 Figure 33. High Standard for P F O S A ................... 88 Figure 34. H igh Standard for M 5 5 6 ........................................................................................................... 89 Figure 35. High Standard for M 5 7 0 ........................................................................................................... 90 Page 7 Northwest Bioanalytical Study No. NWBSOO-091 Report No. NWBRO1-065 Quantitative Determination of PFOS and Related Compounds in Human Serum and Plasma by LC/MS/MS Sample Analysis Report for Protocol EPI-0012 1. INTRODUCTION This report summarizes the analytical results from the quantitation o f PFOS, PFOA, PFHS, PFOSAA, PFOSA, M556, and M 570 in human serum and plasma samples for 3M Company (3M) in support o f Protocol EPI-0012 [5.1]. The LC/MS/MS m ethod for the analytes has a targeted calibration curve range o f 1.00 to 500 ppb (except for M 556 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 n aly te PFOS PFOA PFHS PFOSAA PFOSA M556 M570 LLOQ 3.94 ppb 1.92 ppb 1.36 ppb 1.60 ppb 1.00 ppb 2.50 ppb 1.00 ppb ULOQ 414 ppb 481 ppb 523 ppb 501 ppb 500 ppb 500 ppb 500 ppb The sponsor w as 3M Company (Epidemiology, M edical Department, 220-3W -05, St. Paul, M N 55144), and the Study Director was Jeffrey H. Mandel, M.D., M .P.H. The Study Contact at 3M was Jean Burris. James K. Lundberg was the technical contact at 3M. The following is a list o f NW B 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 study included Suzanne Page 8 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Newman, B.S. (NWB Research Scientist); Em ily Yardimci, B.S. (NWB Associate Research Scientist). NW B 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: 24-Sep-2000 Date Analyses Completed: 07-Apr-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 of the U.S. FDA Good Laboratory Practice Regulations for N onclinical 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). A ny 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. METHODOLOGY The assay used for this study is reported in Northwest Bioanalytical reports NW BR00108 [5.2] and NW BR00-122 [5.3]. Chinese hum an plasm a was used for the calibration curves and quality control samples. Serum sample results obtained using plasma curves are discussed in NWBR00-122. Samples for this study were received at NW B on the following dates: Receipt Date 25-Jul-2000 26-Jul-2000 Number o f Samples Received 243 151 Storage Condition (except during analysis) -20 C -20 C Page 9 Northwest Bioanalytical Reference Material Study No. NWBS00-091 Report No. NWBR01-065 Analyte PFOS PFOA PFHS PFOSAA PFOSA Lot Number Purity Expiration Date Source 193 100% 31-D ec-2010 3M 245 100% 31-D ec-2010 3M SE-036 91.1% 01-Jan-2010 3M 617 53.8% 31-D ec-2010 3M 214 100% 31-D ec-2010 3M *Stored dry Storage Conditions Room temperature* Room temperature* -20 C Room temperature Room temperature Analyte M556 M570 THPFOS Lot Number Purity Expiration Date NB113047-8D 99.89% 31-D ec-2010 118506-26 99.75% 31-D ec-2010 59909 90% 31-D ec-2010 Source 3M 3M 3M Storage Conditions 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 NW B stock solutions used to prepare the analytical standards and controls used for this studies. All arrangements for purity determinations and the transfer of NW B solutions to Centre Analytical Laboratories, Inc., were performed by 3M. Based on Centre Analytical Laboratories' results, the concentrations of plasma calibrator and quality control samples were corrected using the following factors: Analyte PFOS PFOA PFHS Correction Factor 0.821 0.961 1.043 Page 10 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Eight or more calibration standards were prepared on the day of each run by adding 100 (iL o f blank Chinese hum an plasm a and 400 jiL o f 50 m M ammonium acetate in w ater (unadjusted pH --6.9) to 13 x 100 m m polypropylene tubes. After a b rie f vortex m ixing, 10.0 p.L o f the appropriate spiking solution was added. The final calibration standard concentrations in human plasm a for each analyte are listed below: A n aly te__________________ C a lib ra tio n S ta n d a rd C oncen tratio n s* _______________ PFOS 3.94, 5.17, 11.3, 23.6, 44.2, 85.4, 209, 332 and414ppb PFOA 1.92,3.36, 10.6, 25.0,49.0, 97.1, 241, 385 and 481 ppb PFHS 1.36, 2.92, 10.7,26.5, 52.6, 104, 262,418 and 523 ppb PFOSAA 1.60, 3.10, 10.6,25.6, 5 0 .6 ,1 0 1 ,2 5 1 ,4 0 1 and 501 ppb PFOSA 1.00, 2.50, 10.0, 25.0, 50.0, 100, 250,400 and 500 ppb M 556 2.50, 10.0, 25.0, 50.0, 100, 250, 400 and 500 ppb (runs 1 - 17) 3.20, 10.7, 25.7, 50.7, 101, 251,401 and 501 ppb (run 18) M 570 1.00, 2.50, 10.0, 25.0, 50.0, 100, 2 5 0 ,4 0 0 and 500 ppb (runs 1 - 17) __________________ 1.90, 3.40, 10.9, 25.9, 50.9, 101, 251, 401 and 501 ppb (run 18) * The target calibration curve range is 1.00 ppb to 500 ppb (except M556 which has a LLOQ of 2.50 ppb). Each analyte has a different final curve range based upon the persistent levels of the analyte in the human plasma used. Run 18 for M556 and M570 used a new pool o f plasma with different persistent levels of analyte. 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 hum an plasm a on October 6, 2000, October 11, 2000, October 27, 2000 and M arch 8, 2001 and stored in a -20 C freezer. For each run, analytical QC levels were assayed in duplicate. A n aly te PFOS PFOA PFHS PFOSAA PFOSA M556 M570 Low 6.40 4.81 4.48 4.60 4.00 4.00 4.00 Q C C oncentrations (ppb)* M edium 126 145 156 151 150 150 150 High 332 385 417 401 400 400 400 * The target QC concentrations are 4.00 ppb, 150 ppb and 400 ppb. Each analyte has different QC concentrations based upon the persistent levels of the analyte in the human plasma used. Page 11 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 The internal standard (THPFOS) was added to all serum samples (except Blanks) for a final concentration o f approximately 200 ppb. The analytical method consisted o f a liquid:liquid 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 TurboIonSprayTM ionization. 3. SAM PLE ANALYSIS PFOS, PFOA, PFHS, PFOSAA, PFOSA, M556, M570, and THPFOS chromatographic peaks were integrated using the M acQuan software (version 1.6) with a smooth factor o f one. Quantitation was based upon quadratic regression analysis o f w eighted (l/x 2) 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 C riteria For an analytical run to be accepted, it must have m et 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 m ust be w ithin 20% ( 25% for LLOQ) Page 12 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 o f 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. Lower Limit o f Quantitation The back-calculated concentrations o f at least one o f the duplicate lowest points in the calibration curve must be within 25% of the target concentration for PFOSA and PFOSAA to qualify as the LLOQ and within 20% o f the target concentration to qualify as the LLOQ for all other analytes. I f 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 of 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 m ay vary on average up to 26% from results obtained using human serum calibration curves at some concentration levels for PFOA, PFHS, PFOSAA, M556, and M570. For PFOSA, results may vary on average up to 43% from results obtained using human serum calibration curves. This data is reported in Assay Revalidation Addendum Report NW BR00-122 [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. Page 13 Northwest Bioanalytcal Study No. NWBSOO-091 Report No. NWBRO1-065 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 1 Accepted 2 Accepted 3 Accepted 4 Accepted 5 Accepted 6 N/A 7 Accepted 8 Accepted 9 Accepted 10 Accepted 11 12 N/A 13 N/A 14 N/A 15 Accepted 16 Rejected 17 Accepted 18 N/A PFOA Result Rejected Accepted Accepted Accepted Accepted Accepted Accepted Accepted Accepted Accepted N/A N/A N/A Accepted Rejected Accepted N/A PFHS Result Accepted Accepted Accepted Accepted Accepted N/A Accepted Accepted Accepted Accepted N/A N/A N/A Accepted Rejected Accepted N/A PFOSAA Result Comments Accepted Calibration curve failed for rejected analyte. Accepted Pre-study assay evaluation. Accepted Accepted Accepted N/A Accepted Accepted Accepted Accepted Run mistakenly created in Watson DMLIMS system (no data collected). N/A Reinjection of run 7. N/A Reinjection of run 8. N/A Accepted Rejected Rejected due to sample preparation error. Accepted N/A N/A = not analyzed Page 14 Northwest Bioanalytical Run PFOSA Result ID 1 Accepted 2 Accepted 3 Accepted 4 Accepted 5 Accepted 6 N/A 7 Accepted 8 Accepted 9 Accepted 10 Accepted 11 12 N/A 13 N/A 14 N/A 15 Accepted 16 Rejected 17 Accepted 18 N/A Study No. NWBS00-091 Report No. NWBRO1-065 M556 Result M570 Result Comments Accepted Accepted Accepted Accepted Accepted N/A Rejected Rejected Accepted Accepted Accepted Accepted Accepted Accepted Accepted N/A Rejected Accepted Accepted Accepted Accepted Rejected Accepted N/A Accepted Rejected N/A Accepted Accepted Rejected Rejected Rejected Accepted Accepted N/A = not analyzed Pre-study assay evaluation. QCs failed for rejected analytes. Calibration curve failed for rejected analyte. Run mistakenly created in Watson DMLIMS system (no data collected). Reinjection of run 7. QCs failed for rejected analytes. Reinjection of run 8. Rejected due to sample preparation error. QCs failed for rejected analytes. One o f the m edium QC replicates in R un 17 gave very high PFO SA and PFOSAA results. These results appear to be outliers which skew the overall study statistics. W ithout inclusion o f this QC result, the medium QC statistics are as follows: Mean S.D. %CV "/Theoretical %Bias N PFOSAA 162 18.5 11.0 107.3 7.3 19 PFOSA 153 17.7 11.6 102.0 2.0 19 Page 15 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBR01-065 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-0012. "Serum fluorochemical trends o f out-of-country residents in CLUE I (1974) and CLUE II (1989) epidemiologic investigations." 5.2. C. Sakashita. "Quantitative Determination of PFOS, PFOSA, PFOSAA, POAA, PFHS, M556 and M 570 in Human Serum by LC/MS/MS." Assay Revalidation Report. NW B Study NW BS00-040. NW B Report N W B R 00-108. January 24, 2001. 5.3. C. Sakashita. "Quantitative Determination o f PFOS, PFOSA, PFOSAA, POAA, PFHS, M 556 and M 570 in Human Serum by LC/MS/MS." Assay Revalidation Addendum Report. NW B Study NW B S00-040. NW B Report N W B R 00-122. November 20, 2001. 6. D A TA R E T E N T IO N The raw data and final report for this study will be stored in the N W B 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 o f records at completion of contract obligations. Page 16 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Table 1. Calibration Curve Summary for PFOS Quadratic weighted 1/x2. All concentrations are expressed as ppb. R un Date 13-Oct-2000 14-0ct-2000 16 - O c t- 2 0 0 0 18-Oct-2000 19-Oct-2000 20-0ct-2000 20-0ct-2000 23-Oct-2000 31 -Oct-2000 23-Mar-2001 Run N um ber 1 3 5 4 9 7 8 10 15 17 A B C R-Squared LLOQ ULOQ -0.000005 0.029065 0.003518 -0.000007 0.030835 -0.004262 -0.000007 0.024230 -0.022998 -0.000010 0.028656 0.010100 -0.000002 0.020051 0.008280 -0.000008 0.021563 0.013487 -0.000009 0.021893 0.002918 -0.000010 0.022789 0.007538 -0.000005 0.027791 0.010909 0.000002 0.039134 0.020796 0.9970 0.9967 0.9930 0.9911 0.9944 0.9881 0.9922 0.9836 0.9956 0.9869 3.94 414 3.94 414 3.94 414 3.94 414 3.94 414 3.94 414 3.94 414 3.94 414 3.94 414 3.94 414 Mean S.D. %CV n -0.000006 0.000004 -66.7 10 0.026601 0.005748 21.6 10 0.005029 0.011906 236.7 10 0.9919 0.0045 0.5 10 A, B, and C are coefficients used to define the calibration curve. Page 17 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Table 2. Calibration Curve Summary for PFOA Quadratic weighted 1/x2. All concentrations are expressed as ppb. R un Date 14-0ct-2000 15 - O c t- 2 0 0 0 16 - O c t- 2 0 0 0 18-Oct-2000 19 - O c t- 2 0 0 0 20-0ct-2000 20-0ct-2000 23-Oct-2000 31-Oct-2000 23-Mar-2001 Run N um ber 3 6 5 4 9 7 8 10 15 17 A B C R-Squared LLOQ ULOQ 0.000000 0.021802 -0.010687 0.000002 0.022043 -0.000611 0.000000 0.013756 -0.007270 -0.000001 0.015425 -0.002433 0.000004 0.015292 0.002113 0.000000 0.014501 -0.003931 0.000001 0.013316 -0.003358 0.000001 0.013509 0.001640 0.000001 0.018750 0.001244 0.000001 0.008393 0.007706 0.9983 0.9982 0.9982 0.9964 0.9875 0.9979 0.9964 0.9856 0.9967 0.9842 1.92 481 1.92 481 1.92 481 1.92 481 1.92 481 1.92 481 1.92 481 1.92 481 1.92 481 1.92 481 Mean S.D. %cv n 0.000001 0.000001 100.0 10 0.015679 -0.001559 0.004165 0.005206 26.6 -333.9 10 10 0.9939 0.0057 0.6 10 A, B, and C are coefficients used to define the calibration curve. Page 18 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Table 3. Calibration Curve Summary for PFHS Quadratic weighted 1/x2. All concentrations are expressed as ppb. Run Date 13-Oct-2000 14-0ct-2000 16-Oct-2000 18-Oct-2000 19 - O c t- 2 0 0 0 20-0ct-2000 20-0ct-2000 2 3-O ct-2000 31-Oct-2000 2 3-M ar-2001 Run N um ber 1 3 5 4 9 7 8 10 15 17 A B C R-Squared LLOQ ULOQ -0.000010 0.030430 0.000781 -0.000010 0.031509 0.004692 -0.000009 0.027185 -0.000974 -0.000007 0.026276 0.001867 -0.000005 0.026618 0.007959 -0.000006 0.027614 0.003405 -0.000005 0.024990 0.005704 -0.000004 0.024289 0.010779 -0.000009 0.031737 0.006465 -0.000002 0.021861 0.020425 0.9973 0.9958 0.9926 0.9986 0.9895 0.9949 0.9975 0.9948 0.9976 0.9959 1.36 523 1.36 523 1.36 523 1.36 523 1.36 523 1.36 523 1.36 523 1.36 523 1.36 523 1.36 523 Mean S.D. %CV n -0.000007 0.027251 0.000003 0.003211 -42.9 11.8 10 10 0.006110 0.006113 100.0 10 0.9955 0.0027 0.3 10 A, B, and C are coefficients used to define the calibration curve. Page 19 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Table 4. Calibration Curve Summary for PFOSAA Quadratic weighted 1/x2. All concentrations are expressed as ppb. Run Date 13-0ct-2000 14-0ct-2000 16-0ct-2000 18-Oct-2000 19-0ct-2000 20-0ct-2000 20-0ct-2000 23-Oct-2000 31 -Oct-2000 23-Mar-2001 Run N um ber 1 3 5 4 9 7 8 10 15 17 A B C R-Squared LLOQ ULOQ 0.000005 0.026658 0.002620 0.000004 0.033575 -0.001462 0.000000 0.025009 -0.008698 0.000001 0.036355 -0.011781 0.000002 0.021746 0.017518 -0.000004 0.030680 0.001307 -0.000005 0.035370 -0.003269 -0.000003 0.025921 0.008516 -0.000001 0.023946 -0.003099 0.000001 0.007894 0.004964 0.9900 0.9859 0.9925 0.9936 0.9772 0.9809 0.9912 0.9750 0.9952 0.9685 1.60 501 1.60 501 1.60 501 1.60 501 1.60 501 1.60 501 1.60 501 1.60 501 1.60 501 1.60 501 Mean S.D. %CV n 0.000000 0.000003 10 0.026715 0.008305 31.1 10 0.000662 0.008466 1278.9 10 0.9850 0.0091 0.9 10 A, B, and C are coefficients used to define the calibration curve. Page 20 Northwest Bioanalytical Study No. NWBSOO-091 Report No. NWBRO1-065 Table 5. Calibration Curve Summary for PFOSA Quadratic weighted 1/x2. All concentrations are expressed as ppb. R un Date 13-0ct-2000 14-0ct-2000 16-Oct-2000 18-Oct-2000 19-Oct-2000 20-0ct-2000 20-0ct-2000 23-Oct-2000 31 -Oct-2000 23-Mar-2001 Run N um ber 1 3 5 4 9 7 8 10 15 17 A B C R-Squared LLOQ ULOQ -0.000005 0.059585 0.013519 -0.000005 0.060205 0.017572 -0.000004 0.047126 -0.000891 -0.000010 0.071207 0.006196 0.000000 0.036297 0.011607 -0.000007 0.041996 0.014850 -0.000010 0.046357 0.008887 -0.000013 0.044171 0.009049 -0.000013 0.052859 0.003517 0.000002 0.034802 0.013699 0.9914 0.9940 0.9938 0.9853 0.9891 0.9813 0.9868 0.9824 0.9949 0.9847 1.00 500 1.00 500 1.00 500 1.00 500 1.00 500 1.00 500 1.00 500 1.00 500 1.00 500 1.00 500 Mean S.D. %cv n -0.000006 0.049461 0.000005 0.011496 -83.3 23.2 10 10 0.009801 0.005641 57.6 10 0.9884 0.0050 0.5 10 A, B, and C are coefficients used to define the calibration curve. Page 21 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Table 6. Calibration Curve Summary for M556 Quadratic weighted 1/x2. All concentrations are expressed as ppb. R un Date 13-O c t-2 0 0 0 14-0ct-2000 16-0ct-2000 18-Oct-2000 19-0ct-2000 21-Oct-2000 22-Oct-2000 23-Oct-2000 31-Oct-2000 07-Apr-2001 Run N um ber 1 3 5 4 9 13 12 10 15 18 A B C R-Squared LLOQ ULOQ 0.000002 0.018046 0.011395 0.9826 2.50 500 0.000000 0.025056 0.002464 0.9875 2.50 500 -0.000002 0.019822 -0.004705 0.9931 2.50 500 -0.000004 0.027460 -0.000627 0.9882 2.50 500 0.000000 0.021846 0.010790 0.9800 2.50 500 -0.000006 0.029287 0.003549 0.9936 2.50 500 -0.000006 0.026339 0.005660 0.9846 2.50 500 -0.000009 0.025551 -0.002177 0.9893 2.50 500 -0.000001 0.018373 0.001923 0.9972 2.50 500 0.000001 0.007457 0.002028 0.9924 3.20 501 Mean S.D. %cv n -0.000002 0.021924 0.000004 0.006400 -200.0 29.2 10 10 0.003030 0.005167 170.5 10 0.9889 0.0054 0.5 10 A, B, and C are coefficients used to define the calibration curve. Page 22 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Table 7. Calibration Curve Summary for M570 Quadratic weighted 1/x2. All concentrations are expressed as ppb. Run Date 13-Oct-2000 14-0ct-2000 16 - O c t- 2 0 0 0 18-Oct-2000 19-Oct-2000 20-0ct-2000 2 3-Oct-2000 30-0ct-2000 31-Oct-2000 07-Apr-2001 Run N um ber 1 3 5 4 9 8 10 14 15 18 A B C R-Squared LLOQ ULOQ -0.000004 0.044503 0.006965 -0.000007 0.055032 0.008879 -0.000008 0.039426 0.001905 -0.000012 0.056127 0.004675 0.000001 0.035531 0.023102 -0.000017 0.055069 0.020659 -0.000006 0.040644 0.023177 -0.000004 0.033297 0.021686 -0.000006 0.038561 0.012040 -0.000001 0.034524 0.014973 0.9918 0.9884 0.9946 0.9929 0.9784 0.9948 0.9892 0.9878 0.9924 0.9858 1.00 500 1.00 500 1.00 500 1.00 500 1.00 500 1.00 500 1.00 500 1.00 500 1.00 500 1.90 501 Mean S.D. %cv n -0.000006 0.043271 0.013806 0.000005 0.008970 0.008054 -83.3 20.7 58.3 10 10 10 0.9896 0.0049 0.5 10 A, B, and C are coefficients used to define the calibration curve. Page 23 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBR01-065 Table 8. Back-Calculated Concentrations of Calibration Standards for PFOS Quadratic weighted 1/x2. All concentrations are expressed as ppb. Run Date 13-Oct-2000 14-0ct-2000 16-0ct-2000 18-Oct-2000 19-0ct-2000 20-0ct-2000 20-0ct-2000 23-Oct-2000 31 -Oct-2000 23-Mar-2001 Run Number 1 3 5 4 9 7 8 10 15 17 3.94 5.17 3.81 3.81 3.83 *7.35 4.08 3.65 4.30 3.54 4.30 3.80 3.32 4.67 3.76 3.73 3.80 4.28 3.65 4.10 4.01 3.42 5.60 5.08 4.87 5.33 5.12 5.16 5.63 4.72 4.92 5.23 4.82 5.42 4.79 6.03 4.56 5.50 5.67 4.90 5.70 5.60 11.3 23.6 44.2 85.4 209 332 414 12.2 24.2 43.7 86.4 209 331 415 11.2 22.1 45.1 77.4 220 336 406 11.6 24.1 43.1 83.3 203 336 428 12.5 24.9 42.5 77.5 215 325 414 11.3 23.1 42.6 71.2 201 324 406 12.9 24.6 45.8 90.2 200 363 427 12.5 26.1 46.6 85.9 218 350 442 10.2 22.3 42.3 78.6 199 316 392 10.3 24.1 50.5 78.2 213 342 410 10.8 24.2 46.3 78.8 212 336 403 10.0 21.7 44.2 77.9 212 322 395 12.1 24.7 49.8 85.4 219 334 430 11.3 23.2 41.8 76.7 200 327 431 11.8 26.1 47.7 80.9 204 333 434 9.95 20.7 53.2 77.5 *122 307 371 12.5 *3.05 44.9 89.6 197 365 464 12.0 23.4 45.0 84.2 224 347 424 10.7 23.3 42.3 84.5 203 345 368 10.6 26.0 41.2 81.4 232 293 419 11.3 20.3 48.5 89.2 186 380 399 Mean S.D. %cv %Bias n 3.89 5.23 11.4 23.6 45.4 81.7 209 336 414 0.337 0.401 0.913 1.68 3.24 5.11 11.1 20.0 23.0 8.7 7.7 8.0 7.1 7.1 6.3 5.3 6.0 5.6 -1.3 1.2 0.9 0.0 2.7 -4.3 0.0 1.2 0.0 19 20 20 19 20 20 19 20 20 * Sample deactivated as an outlier (not included in summary statistics) Page 24 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBR01-065 Table 9. Back-Calculated Concentrations of Calibration Standards for PFOA Quadratic weighted 1/x2. All concentrations are expressed as ppb. Run Date 14-0ct-2000 15-0ct-2000 16-0ct-2000 18-Oct-2000 19-0ct-2000 20-0ct-2000 20-0ct-2000 23-Oct-2000 31 -Oct-2000 23-Mar-2001 Run Number 3 6 5 4 9 7 8 10 15 17 1.92 3.36 1.97 *7.07 1.95 1.89 1.99 1.85 1.74 2.16 2.02 2.12 1.77 2.05 1.99 1.88 2.04 2.00 1.94 1.88 1.54 2.40 3.36 3.12 3.43 3.34 3.33 3.33 3.25 3.32 2.83 3.08 3.52 3.28 2.92 3.62 3.22 2.81 3.49 3.36 2.87 3.40 10.6 25.0 49.0 97.1 241 385 481 10.5 25.3 50.2 94.4 240 388 470 11.4 25.9 46.4 92.6 248 396 480 9.83 25.6 46.9 97.3 245 380 461 10.4 26.5 50.2 92.8 256 396 480 10.9 25.7 48.8 89.6 241 402 483 10.5 26.1 50.7 92.5 233 402 461 9.97 25.7 47.0 93.5 236 403 466 10.5 25.9 52.6 97.4 249 386 475 8.96 23.9 53.8 81.6 244 371 454 10.4 28.5 55.4 98.6 270 403 478 9.98 25.5 50.0 92.7 243 371 478 10.7 24.7 52.0 95.1 244 384 497 10.7 25.8 48.1 94.6 243 392 493 11.0 26.7 50.0 88.7 226 382 485 10.9 25.1 *69.9 110 173 408 476 10.8 *10.2 48.9 108 225 414 493 9.80 24.9 47.2 89.3 250 383 470 10.7 26.6 49.5 99.2 247 427 443 12.5 27.4 50.2 90.7 256 383 492 10.7 20.7 48.5 99.6 222 400 465 Mean S.D. %cv %Bias n 1.96 0.178 9.1 2.1 19 3.24 0.234 7.2 -3.6 20 10.6 0.705 6.7 0.0 20 25.6 1.56 6.1 2.4 19 49.8 94.9 240 394 475 2.39 6.41 19.3 14.3 14.0 4.8 6.8 8.0 3.6 2.9 1.6 -2.3 -0.4 2.3 -1.2 19 20 20 20 20 * Sample deactivated as an outlier (not included in summary statistics) Page 25 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBR01-065 Table 10. Back-Calculated Concentrations of Calibration Standards for PFHS Quadratic weighted 1/x2. All concentrations are expressed as ppb. Run Date 13-0ct-2000 Run Number 1 14-0ct-2000 3 16-0ct-2000 5 18-Oct-2000 4 19-0ct-2000 9 20-0ct-2000 7 20-0ct-2000 8 23-Oct-2000 10 31-Oct-2000 15 23-Mar-2001 17 1.36 2.92 1.25 1.37 1.41 *7.41 1.56 1.15 1.38 *25.7 1.39 1.23 1.21 1.59 1.41 1.31 1.34 1.47 1.30 1.42 1.41 1.31 3.25 3.03 2.95 2.56 2.99 2.86 2.93 2.81 2.93 3.45 2.75 2.72 2.90 2.90 2.68 2.76 3.01 2.77 3.13 2.71 10.7 26.5 52.6 104 262 418 523 10.7 25.9 51.1 101 248 398 528 11.1 26.2 52.6 99.7 272 431 543 10.6 27.9 52.6 104 251 418 540 12.2 26.6 52.2 94.7 254 418 539 10.7 27.5 51.0 92.8 257 413 528 10.9 26.5 50.7 120 245 439 524 10.6 29.0 51.6 109 252 426 537 10.4 26.0 53.2 101 257 411 522 8.79 25.0 55.8 84.1 255 430 511 11.5 27.8 56.1 106 278 421 524 11.0 27.8 56.3 101 269 408 510 10.1 26.5 53.5 101 264 423 531 11.4 28.6 52.9 102 259 429 557 10.4 26.9 53.1 93.1 248 411 516 10.7 27.9 *74.5 115 *176 432 502 10.1 *8.57 52.1 110 231 431 531 11.2 26.0 49.7 98.7 268 419 530 11.2 27.4 52.0 102 266 455 478 10.6 28.1 52.5 98.5 283 405 527 10.8 28.5 48.1 104 237 449 512 Mean S.D. %CV %Bias n 1.36 0.114 8.4 0.0 18 2.90 0.208 7.2 -0.7 20 10.7 0.677 6.3 0.0 20 27.2 1.09 4.0 2.6 19 52.5 102 258 423 525 2.06 7.95 13.3 14.3 16.9 3.9 7.8 5.2 3.4 3.2 -0.2 -1.9 -1.5 1.2 0.4 19 20 19 20 20 * Sample deactivated as an outlier (not included in summary statistics) Page 26 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBR01-065 Table 11. Back-Calculated Concentrations of Calibration Standards for PFOSAA Quadratic weighted 1/x2. All concentrations are expressed as ppb. Run Date Run Number 1.60 3.10 10.6 25.6 50.6 101 251 401 501 13-Oct-2000 1 1.54 3.29 10.4 24.7 45.5 91.3 236 383 472 1.50 3.50 11.2 25.1 54.2 92.6 308 425 502 14-0ct-2000 3 1.61 2.40 10.9 24.2 45.0 93.4 237 388 494 *5.30 3.55 12.9 27.7 46.9 98.3 292 405 506 16-0ct-2000 5 1.84 3.05 11.0 28.0 52.0 89.1 263 428 484 1.42 2.85 10.9 24.4 50.8 95.8 247 440 457 18-0ct-2000 4 1.45 2.81 10.1 23.1 49.2 93.8 250 384 472 1.73 3.49 10.6 26.1 53.7 110 269 424 506 19-Oct-2000 9 1.58 2.29 7.91 23.7 52.4 89.8 248 371 450 1.74 3.61 10.4 29.1 62.5 100 287 435 516 20-0ct-2000 7 1.31 4.10 10.5 24.8 57.6 99.0 *373 425 514 1.71 2.84 10.4 22.0 51.5 87.6 275 380 472 20-0ct-2000 8 1.72 3.26 10.7 26.6 53.1 102 291 424 536 1.40 3.34 9.16 26.8 46.5 94.0 237 356 469 23-Oct-2000 10 1.91 2.72 10.9 24.5 68.5 104 179 424 502 1.52 2.56 10.6 *1.44 49.1 102 238 409 540 31 -Oct-2000 15 1.41 3.26 10.7 24.0 48.1 94.6 271 394 487 1.68 3.36 10.9 25.4 48.1 106 261 433 467 23-Mar-2001 17 *7.76 4.18 10.6 24.9 46.5 94.5 301 321 525 - 1.32 3.23 10.3 17.3 59.0 108 243 459 460 Mean S.D. %CV %Bias n 1.58 3.18 10.6 24.9 52.0 97.3 260 405 492 0.176 0.504 0.916 2.54 6.06 6.42 30.3 33.1 26.6 11.1 15.8 8.6 10.2 11.7 6.6 11.7 8.2 5.4 -1.3 2.6 0.0 -2.7 2.8 -3.7 3.6 1.0 -1.8 18 20 20 19 20 20 19 20 20 * Sample deactivated as an outlier (not included in summary statistics) Page 27 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Table 12. Back-Calculated Concentrations of Calibration Standards for PFOSA Quadratic weighted l/x2. All concentrations are expressed as ppb. Run Date 13-Oct-2000 14-0ct-2000 16-Oct-2000 18-Oct-2000 19-0ct-2000 20-0ct-2000 20-0ct-2000 23-Oct-2000 31 -Oct-2000 23-Mar-2001 Run 1.00 2.50 10.0 25.0 50.0 100 250 400 500 Number 1 1.01 2.84 10.6 25.8 46.6 93.0 249 390 510 0.864 2.91 10.2 22.4 49.3 87.2 275 408 493 3 0.980 2.21 10.3 25.8 45.8 98.5 242 375 513 *2.36 2.87 10.3 27.1 46.0 94.0 277 395 510 5 1.06 2.43 9.13 24.5 48.3 86.9 249 394 472 0.924 2.66 11.2 25.6 54.0 97.1 255 458 485 4 1.07 2.76 12.6 26.3 54.4 98.8 285 417 508 0.936 2.17 8.44 21.9 46.6 89.8 246 386 469 9 1.15 2.36 8.79 25.8 59.2 83.0 268 399 496 0.882 2.52 9.47 27.1 53.8 89.7 260 410 480 7 0.760 2.34 8.72 23.1 48.2 90.2 247 371 456 1.24 2.70 11.4 22.8 59.1 105 281 405 541 8 0.851 2.21 9.55 22.1 44.0 90.8 256 391 479 1.11 3.05 10.3 29.2 50.9 106 259 396 528 10 0.939 2.19 9.62 22.7 55.0 99.1 168 392 477 1.08 2.60 11.5 *1.08 55.8 102 227 474 573 15 0.930 2.62 9.96 25.7 52.8 101 289 438 529 1.06 2.48 9.45 24.1 47.2 100 229 362 453 17 0.984 2.94 9.66 26.3 50.7 93.2 285 370 522 0.906 2.80 10.6 17.0 49.6 106 240 431 458 Mean S.D. %cv %Bias n 0.986 2.58 10.1 24.5 50.9 95.6 254 403 498 0.117 0.276 1.04 2.70 4.46 6.76 27.5 28.9 31.4 11.9 10.7 10.3 11.0 8.8 7.1 10.8 7.2 6.3 -1.4 3.2 1.0 -2.0 1.8 -4.4 1.6 0.8 -0.4 19 20 20 19 20 20 20 20 20 * Sample deactivated as an outlier (not included in summary statistics) Page 28 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 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 3.20 10.0 10.7 25.0 25.7 50.0 50.7 100 101 250 251 400 401 500 501 Number 13-0ct-2000 1 2.06 9.64 23.4 43.9 88.0 231 358 449 2.89 11.3 26.2 55.6 97.1 301 450 526 14-0ct-2000 3 1.97 10.4 23.8 43.8 94.1 236 394 491 2.93 11.4 28.1 47.3 97.5 277 395 522 16-0ct-2000 5 2.53 10.8 28.1 52.4 90.7 272 414 498 2.41 10.2 22.9 49.1 88.6 230 440 464 18-0ct-2000 4 2.15 9.36 23.4 46.1 89.8 228 375 455 2.89 9.85 26.1 56.9 112 274 438 526 19-0ct-2000 9 2.15 8.10 23.1 52.0 78.5 237 376 446 2.93 9.93 29.5 59.4 101 273 433 544 21-Oct-2000 13 2.14 9.49 25.2 48.2 92.8 234 389 500 2.87 10.1 26.9 54.0 98.1 257 427 499 22-Oct-2000 12 2.73 10.2 26.4 58.2 102 299 445 577 2.27 9.80 22.0 49.5 84.0 223 342 429 23-Oct-2000 10 2.60 10.0 22.8 62.8 97.1 *157 398 484 2.43 9.42 *1.95 49.6 99.2 211 413 547 * Sample deactivated as an outlier (not included in summary statistics) Page 29 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBR01-065 Table 13. Back-Calculated Concentrations of Calibration Standards for M556 (Continued) Quadratic weighted 1/x2. All concentrations are expressed as ppb. Run Date Run 2.50 3.20 10.0 10.7 25.0 25.7 50.0 50.7 100 101 250 251 400 401 500 501 Number 31-Oct-2000 15 2.33 9.67 24.3 51.9 95.2 255 388 499 2.70 9.81 25.8 49.3 104 249 438 470 07-Apr-2001 18 3.47 9.97 28.4 49.5 99.9 258 425 478 2.95 11.4 20.9 54.3 104 239 427 477 Mean S.D. %cv %Bias n 2.50 3.21 9.97 10.7 25.2 24.7 51.7 51.9 95.0 102 252 249 406 426 496 478 0.331 ISD 0.747 ISD 2.19 ISD 5.32 ISD 7.79 ISD 26.7 ISD 31.8 ISD 39.7 ISD 13.2 ISD 7.5 ISD 8.7 ISD 10.3 ISD 8.2 ISD 10.6 ISD 7.8 ISD 8.0 ISD 0.0 0.3 -0.3 0.0 0.8 -3.9 3.4 2.4 -5.0 1.0 0.8 -0.8 1.5 6.2 -0.8 -4.6 18 2 18 2 17 2 18 2 18 2 17 2 18 2 18 2 * Sample deactivated as an outlier (not included in summary statistics) ISD = insufficient data points for statistical calculations Northwest Bioanalytical Study No. NWBSOO-091 Report No. NWBRO1-065 Table 14. Back-Calculated Concentrations of Calibration Standards for M570 Quadratic weighted 1/x2. All concentrations are expressed as ppb. Run Date 13-0ct-2000 14-0ct-2000 16-Oct-2000 18-Oct-2000 19-0ct-2000 20-0ct-2000 23-Oct-2000 30-0ct-2000 31 -Oct-2000 Run Number 1 3 5 4 9 8 10 14 15 1.00 1.90 2.50 3.40 10.0 10.9 25.0 25.9 50.0 50.9 100 101 250 251 400 401 500 501 0.918 0.994 1.00 *3.99 1.14 0.900 0.895 1.12 0.939 1.16 0.995 0.985 1.02 1.04 0.936 1.13 0.828 1.18 2.64 2.93 1.99 2.90 2.33 2.35 2.20 2.76 1.78 2.64 2.45 2.68 2.30 2.26 2.35 2.15 2.29 2.66 9.83 24.0 45.7 91.3 236 382 479 10.4 24.1 53.9 91.1 295 423 506 10.4 23.2 44.3 94.0 237 386 498 11.5 27.8 47.0 97.4 286 403 505 10.6 26.4 50.7 89.7 260 409 486 10.6 24.6 50.9 95.5 243 434 479 9.60 22.8 47.5 95.3 245 395 456 10.1 25.9 54.8 106 270 426 503 7.85 23.6 51.5 89.2 240 364 452 10.2 29.3 61.3 103 281 434 525 10.6 26.0 53.0 99.8 274 422 559 9.12 26.2 45.9 90.7 231 365 464 10.4 25.4 *72.1 108 179 430 503 10.6 *1.13 51.2 109 229 402 541 10.8 27.2 49.5 83.1 231 381 491 9.43 27.5 58.8 98.3 224 451 531 10.1 23.6 48.1 93.7 262 392 486 9.94 25.9 50.6 109 252 431 475 * Sample deactivated as an outlier (not included in summary statistics) Page 31 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Table 14. Back-Calculated Concentrations of Calibration Standards for M570 (Continued) Quadratic weighted 1/x2. All concentrations are expressed as ppb. Run Date 07-Apr-2001 Run Number 18 1.00 1.90 2.50 3.40 10.0 10.9 25.0 25.9 50.0 50.9 100 101 250 251 400 401 500 501 1.76 3.34 11.0 30.5 52.3 109 263 436 476 2.22 2.90 10.9 19.5 52.0 102 231 426 466 Mean S.D. %CV %Bias n 1.01 1.99 2.43 3.12 10.1 11.0 25.5 25.0 50.9 52.2 96.9 106 249 247 407 431 497 471 0.104 ISD 0.309 ISD 0.788 ISD 1.83 ISD 4.55 ISD 7.57 ISD 27.5 ISD 25.6 ISD 28.8 ISD 10.3 ISD 12.7 ISD 7.8 ISD 7.2 ISD 8.9 ISD 7.8 ISD 11.0 ISD 6.3 ISD 5.8 ISD 1.0 4.7 -2.8 -8.2 1.0 0.9 2.0 -3.5 1.8 2.6 -3.1 5.0 -0.4 -1.6 1.8 7.5 -0.6 -6.0 17 2 18 2 18 2 17 2 17 2 18 2 18 2 18 2 18 2 * Sample deactivated as an outlier (not included in summary statistics) ISD = insufficient data points for statistical calculations Page 32 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Table 15. Analytical QC Summary for PFOS All concentrations are expressed as ppb. Run Date Run Number 13-Oct-2000 1 14-0ct-2000 3 16-0ct-2000 5 18-Oct-2000 i4 19-0ct-2000 9 20-0ct-2000 7 20-0ct-2000 8 23-Oct-2000 10 31-Oct-2000 15 23-Mar-2001 17 Low QC (6.40 ppb) 6.41 6.14 6.48 6.80 7.41 *7.81 5.61 5.31 6.77 6.94 *4.41 6.34 5.88 6.28 6.61 5.83 6.62 6.18 *4.60 6.50 Medium QC (126 ppb) 132 127 132 126 109 130 120 107 130 131 131 137 125 139 139 130 129 135 135 *192 High QC (332 ppb) 330 355 339 348 353 309 266 302 340 343 323 375 377 352 359 338 323 322 354 333 Mean S.D. %cv "/(Theoretical %Bias n 6.25 0.826 13.2 97.7 -2.3 20 132 16.5 12.5 104.8 4.8 20 * > 20% deviation from theoretical 337 25.7 7.6 101.5 1.5 20 Page 33 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Table 16. Analytical QC Summary for PFOA All concentrations are expressed as ppb. Run Date 14-0ct-2000 15-0ct-2000 16-0ct-2000 18-Oct-2000 19-0ct-2000 20-0ct-2000 20-0ct-2000 23-Oct-2000 31 -Oct-2000 23-Mar-2001 Run Number Low QC Medium QC (4.81 ppb) (145 ppb) 3 5.08 146 5.18 , 149 6 4.45 147 4.72 143 5 5.77 142 5.53 151 4 4.57 147 4.64 157 9 4.68 152 5.47 155 7 *3.70 150 4.84 172 8 4.83 152 4.87 *181 10 5.13 171 5.28 167 15 4.87 156 4.90 147 17 *2.72 159 4.85 152 High QC (385 ppb) 391 393 359 377 384 374 399 388 390 422 414 398 *466 386 425 431 399 380 412 381 Mean S.D. %CV %Theoretical %Bias n 4.80 0.660 13.8 99.8 -0.2 20 155 10.5 6.8 106.9 6.9 20 * > 20% deviation from theoretical 398 24.3 6.1 103.4 3.4 20 Page 34 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Table 17. Analytical QC Summary for PFHS All concentrations are expressed as ppb. Run Date 13-Oct-2000 14-0ct-2000 16-0ct-2000 18-0ct-2000 19-0ct-2000 20-0ct-2000 20-0ct-2000 23-Oct-2000 31-Oct-2000 23-Mar-2001 Run Number Low QC Medium QC (4.48 ppb) (156 ppb) 1 4.24 149 4.43 139 3 4.60 154 4.69 153 5 5.35 159 4.81 156 4 4.50 160 4.08 161 9 4.66 159 4.83 155 7 4.84 163 5.12 167 8 4.36 157 5.21 *188 10 4.64 172 4.66 178 15 4.35 160 4.60 151 17 4.29 155 4.06 171 High QC (417 ppb) 373 421 392 410 441 386 400 408 415 419 406 387 469 440 425 442 403 400 428 406 Mean S.D. %CV %Theoretical %Bias n 4.62 0.349 7.6 103.1 3.1 20 160 10.8 6.8 102.6 2.6 20 * > 20% deviation from theoretical 414 22.9 5.5 99.3 -0.7 20 Page 35 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Table 18. Analytical QC Summary for PFOSAA All concentrations are expressed as ppb. Run Date 13-Oct-2000 14-0ct-2000 16-0ct-2000 18-0ct-2000 19-0ct-2000 20-0ct-2000 20-0ct-2000 23-Oct-2000 31-Oct-2000 23-Mar-2001 Run Number Low QC Medium QC (4.60 ppb) (151 ppb) 1 5.10 162 5.08 152 3 4.46 176 4.85 148 5 *5.86 136 5.14 156 4 4.08 136 3.98 152 9 *3.38 150 4.43 152 7 *3.10 183 4.26 *194 8 4.73 184 3.91 *201 10 4.24 160 4.29 143 15 4.43 160 4.03 157 17 *2.38 172 4.11 *374 High QC (401 ppb) 411 456 425 456 395 372 379 346 401 426 462 450 *538 377 459 458 393 394 463 456 Mean S.D. %cv %Theoretical %Bias n 4.29 0.772 18.0 93.3 -6.7 20 172 50.8 29.5t 113.9t 13.9t 20 426 45.0 10.6 106.2 6.2 20 * > 25% deviation from theoretical | See comments in Section 4, Results and Discussion Page 36 Northwest Bioanalytical Study No. NWBSO0-091 Report No. NWBRO1-065 Table 19. Analytical QC Summary for PFOSA All concentrations are expressed as ppb. Run Date 13-Oct-2000 14-0ct-2000 16-0ct-2000 18-0ct-2000 19-0ct-2000 20-0ct-2000 20-0ct-2000 23-Oct-2000 31-Oct-2000 23-Mar-2001 Run Number Low QC Medium QC (4.00 ppb) (150 ppb) 1 4.37 148 3.84 143 3 3.72 166 4.21 148 5 4.55 125 4.68 160 4 3.29 127 3.20 *109 9 4.59 170 4.65 171 7 *2.21 150 4.14 158 8 3.64 155 4.08 163 10 4.58 175 4.07 146 15 3.74 154 4.10 158 17 *2.33 180 4.47 *328 High QC (400 ppb) 392 426 420 449 418 375 *280 334 440 432 368 467 467 350 *483 450 423 412 434 417 Mean S.D. %CV %Theoretical %Bias n 3.92 0.712 18.2 98.0 -2.0 20 162 42.9 26.5t 108.0t 8.0 20 412 49.8 12.1 103.0 3.0 20 * > 25% deviation from theoretical f See comments in Section 4, Results and Discussion Page 37 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Table 20. Analytical QC Summary for M556 All concentrations are expressed as ppb. Run Date 13-Oct-2000 14-0ct-2000 16-0ct-2000 18-0ct-2000 19-0ct-2000 21 -Oct-2000 22-Oct-2000 23-Oct-2000 31-Oct-2000 07-Apr-2001 Run Number Low QC Medium QC (4.00 ppb) (150 ppb) 1 3.65 149 4.28 155 3 3.93 150 4.30 156 5 *5.47 143 4.68 162 4 3.81 140 3.96 157 9 4.10 146 4.45 147 13 3.64 145 4.14 167 12 *2.62 *194 3.33 177 10 4.60 157 4.32 140 15 3.95 161 3.93 149 18 4.73 152 3.51 145 High QC (400 ppb) 398 459 452 444 395 378 407 395 387 457 426 411 432 401 443 439 417 406 407 387 Mean S.D. %cv %Theoretical %Bias n 4.07 0.599 14.7 101.8 1.8 20 155 13.1 8.5 103.3 3.3 20 * > 20% deviation from theoretical 417 25.2 6.0 104.3 4.3 20 Page 38 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Table 21. Analytical QC Summary for M570 All concentrations are expressed as ppb. Run Date 13-Oct-2000 14-0ct-2000 16-0ct-2000 18-Oct-2000 19-0ct-2000 20-0ct-2000 23-Oct-2000 30-0ct-2000 31-Oct-2000 07-Apr-2001 Run Number Low QC Medium QC (4.00 ppb) (150 ppb) 1 4.60 157 4.46 148 3 4.13 166 4.47 150 5 *5.02 134 4.75 159 4 3.58 * 135 3.51 150 9 3.85 151 *4.84 157 8 4.37 175 3.68 *207 10 4.20 167 4.43 148 14 3.21 159 3.86 159 15 3.70 164 3.99 160 18 *4.90 159 4.47 156 High QC (400 ppb) 408 454 432 459 392 375 384 352 400 434 *561 372 468 473 402 413 410 416 446 423 Mean S.D. %CV %Theoretical %Bias n 4.20 0.512 12.2 105.0 5.0 20 158 15.2 9.6 105.3 5.3 20 * > 20% deviation from theoretical 424 46.2 10.9 106.0 6.0 20 Page 39 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Table 22. Serum Study Sample Concentrations All concentrations are expressed as ppb. Sample No. 00223 00461 00660 00759 01339 01346 01485 01696 01739 01816 01997 02005 02393 03257 03423 03450 03482 04063 04627 04789 04854 05250 05325 05333 05370 05447 05456 05570 PFOS PFOA* PFHS* PFOSAA* PFOSA* M556* M570* 23.9 <LLOQ(1.92) 1.54 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 16.6 2.47 1.96 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 13.8 2.52 <LLOQ(1.36) 3.12 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 23.0 1.96 <LLOQ(1.36) 1.95 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 16.3 2.79 1.38 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 10.9 <LLOQ(l .92) <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 13.2 <LLOQ(l .92) <LLOQ(l .36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(l .00) 18.6 2.43 1.36 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) 1.22 15.7 <LLOQ(1.92) <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 22.3 2.23 <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 67.8 2.65 3.63 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 55.4 3.02 1.66 12.9 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 25.8 <LLOQ(1.92) 2.36 2.01 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 41.4 2.13 1.46 1.65 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 23.6 3.10 <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 60.1 3.57 3.11 1.86 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 80.2 3.30 4.59 1.79 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 32.8 3.31 3.18 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 44.6 2.98 3.33 3.67 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 25.5 4.53 1.65 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 20.9 2.17 1.75 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 38.9 3.66 2.05 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 56.0 6.57 4.49 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 37.5 2.58 2.97 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 49.0 2.87 2.41 3.78 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 32.4 3.86 2.54 1.71 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 61.9 3.42 2.55 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 50.4 2.50 1.87 1.67 <LLOQ(1.00) <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-091 Report No. NWBR01-065 Table 22. Serum Study Sample Concentrations (Continued) All concentrations are expressed as ppb. Sample No. 05573 05842 05843 10277 10525 10526 10637 10664 10726 10736 10773 10837 10873 10881 10885 10891 10899 10910 10960 10991 11174 11214 11260 12651 13214 13225 13421 13592 PFOS PFOA* PFHS* PFOSAA* PFOSA* M556* M570* 52.1 2.81 2.00 2.14 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 24.3 <LLOQ(1.92) <LLOQ(1.36) 1.67 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 26.3 <LLOQ(l .92) 1.58 1.87 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 27.8 2.01 <LLOQ(1.36) 2.23 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 34.9 2.36 3.01 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 112 3.01 8.41 2.14 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 38.7 2.61 2.87 2.15 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 54.3 4.11 6.17 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 21.4 2.17 <LLOQ(1.36) 2.38 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 82.0 2.54 2.61 6.96 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 42.3 <LLOQ(1.92) 4.05 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 52.7 2.07 3.49 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 35.0 2.31 1.93 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 30.6 <LLOQ(1.92) 1.96 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 29.3 2.49 3.69 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 15.6 <LLOQ(1.92) 1.42 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 24.0 3.11 1.88 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 23.5 <LLOQ(1.92) 1.36 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 34.6 1.97 2.71 2.34 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 26.1 <LLOQ(1.92) 1.50 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 24.6 2.59 2.06 1.83 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 37.6 <LLOQ(1.92) 1.52 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) CLLOQ(I.OO) 139 2.44 6.09 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 19.5 <LLOQ(1.92) <LLOQ(1.36) 2.44 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 22.7 <LLOQ(1.92) 1.63 2.58 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 19.2 1.98 1.50 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 20.0 <LLOQ(1.92) 1.57 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 41.4 4.21 2.51 <LLOQ(1.60) <LLOQ(1.00) <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 P F O S A 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 Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Table 22. Serum Study Sample Concentrations (Continued) All concentrations are expressed as ppb. Sample PFOS No. PFOA* PFHS* PFOSAA* PFOSA* M556* M570* 13657 21.3 <LLOQ(1.92) <LLOQ(1.36) 2.52 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 13738 20.0 3.48 1.67 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 13911 8.76 <LLOQ(1.92) <LLOQ(1.36) <LLOQ(1.60) <LLOQ(l .00) <LLOQ(2.50) <LLOQ(1.00) 14068 29.1 <LLOQ(1.92) 1.96 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 14071 11.6 <LLOQ(1.92) <LLOQ(1.36) <LLOQ(1.60) cLLOQ(l.OO) <LLOQ(2.50) <LLOQ(1.00) 14089 30.8 2.15 1.45 1.74 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 14091 26.0 2.34 2.55 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 14338 33.7 1.93 <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 14357 15.9 2.39 <LLOQ(1.36) 1.83 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 14363 46.8 2.34 3.06 2.45 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 14679 25.5 2.75 1.92 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) 1.12 14726 35.8 2.43 2.25 1.81 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 14863 22.3 2.06 <LLOQ(1.36) 3.28 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 14993 25.1 <LLOQ(1.92) <LLOQ(1.36) 1.83 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 15016 15.7 3.20 2.39 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 15317 24.3 2.17 <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 15508 34.6 2.87 3.24 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 15744 36.3 2.84 2.93 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 15750 14.9 <LLOQ(1.92) <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 15756 16.9 2.73 <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 20198 33.6 3.19 1.52 1.64 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 20219 57.4 6.19 6.33 3.78 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 20554 35.7 2.36 2.28 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 21040 40.2 1.96 1.92 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) 1.84 21476 24.5 2.09 <LLOQ(1.36) 2.00 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 23023 68.7 4.55 4.04 2.52 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 23704 15.0 2.10 <LLOQ(1.36) 1.88 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 23724 23.3 2.59 2.26 <LLOQ(1.60) <LLOQ(1.00) <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 42 Northwest Bioanalytical Study No. NWBSOO-091 Report No. NWBRO1-065 Table 22. Serum Study Sample Concentrations (Continued) All concentrations are expressed as ppb. Sample PFOS No. PFOA* PFHS* PFOSAA* PFOSA* M556* M570* 23957 32.0 3.83 4.08 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 24003 18.0 2.43 1.72 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 24020 32.7 2.28 <LLOQ(1.36) 5.29 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 24022 31.4 2.70 1.65 1.86 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 24077 21.5 2.00 <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 24138 36.4 2.29 2.45 <LLOQ(1.60) <LLOQ(l .00) <LLOQ(2.50) <LLOQ(1.00) 24206 18.4 2.48 <LLOQ(1.36) 1.77 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 24363 35.9 2.11 <LLOQ(1.36) 2.38 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 24413 45.6 3.75 2.29 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 24421 7.14 <LLOQ(1.92) <LLOQ(1.36) 8.21 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 24958 41.4 3.28 1.69 2.06 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 24959 36.8 2.91 <LLOQ(1.36) 1.97 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 24963 46.8 6.11 4.57 3.01 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 24968 51.7 6.36 4.42 3.06 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 30612 20.1 1.94 <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 30713 38.1 2.60 1.57 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) 3.92 31297 36.4 2.52 1.54 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 31432 35.3 3.28 2.00 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 31652 27.1 <LLOQ(1.92) <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 32126 39.6 3.09 2.16 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 32535 13.3 <LLOQ(1.92) 1.75 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 32704 25.1 2.34 <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 33007 45.0 2.97 1.43 1.94 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 33292 23.3 <LLOQ(1.92) <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 33340 25.1 <LLOQ(1.92) <LLOQ(1.36) 5.68 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 33341 20.3 <LLOQ(1.92) 1.43 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 33787 15.8 <LLOQ(1.92) <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 33788 27.1 3.01 <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <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. NWBS00-091 Report No. NWBR01-065 Table 22. Serum Study Sample Concentrations (Continued) All concentrations are expressed as ppb. Sample PFOS No. PFOA* PFHS* PFOSAA* PFOSA* M556* M570* 33793 32.4 2.61 1.40 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 33897 68.2 2.32 2.54 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 33914 147 <LLOQ(1.92) 3.11 3.17 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 33958 36.4 1.97 1.69 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 34087 42.0 2.54 1.86 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 34117 54.8 4.09 <LLOQ(1.36) 2.76 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 34119 141 3.97 2.91 1.77 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 34222 134 3.25 5.17 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 34279 14.3 <LLOQ(1.92) <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 34307 31.7 2.46 3.67 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 34549 178 3.51 10.6 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 34917 68.2 3.45 3.12 3.06 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 35082 29.4 2.09 <LLOQ(l .36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) 4.40 35084 18.3 <LLOQ(1.92) <LLOQ(1.36) <LLOQ(1.60) <LLOQ( 1.00) <LLOQ(2.50) CLLOQ(I.OO) 35174 21.9 2.69 1.81 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 35253 67.6 2.47 5.72 2.00 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 35318 36.1 3.46 <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) 3.44 35332 36.7 2.33 2.00 2.50 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 35346 45.0 3.57 2.00 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 35576 33.1 8.03 1.70 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 35599 34.6 <LLOQ(1.92) 1.44 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 35935 34.7 <LLOQ(1.92) 1.64 2.27 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 35946 35.7 <LLOQ(1.92) 1.72 2.72 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 35950 30.4 2.25 <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 35956 29.0 1.96 <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 40106 59.7 3.99 5.19 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 40388 14.2 <LLOQ(1.92) <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 40734 25.2 <LLOQ(1.92) <LLOQ(1.36) 2.16 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(l .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 Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Table 22. Serum Study Sample Concentrations (Continued) All concentrations are expressed as ppb. Sample No. 40790 40867 40868 40881 40916 40955 40991 40993 41000 41002 41003 41012 41014 41020 41067 41499 41508 41509 41516 41523 41551 41559 41566 41569 41605 41611 41613 41620 PFOS PFOA* PFHS* PFOSAA* PFOSA* M556* M570* 32.5 2.44 <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 26.2 <LLOQ(1.92) 1.75 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 57.9 2.39 3.57 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 29.6 3.47 1.53 1.65 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 15.6 <LLOQ(1.92) <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 9.52 <LLOQ(1.92) <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 20.4 2.16 <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 24.8 <LLOQ(1.92) <LLOQ(1.36) <LLOQ(1.60) <LLOQ(l .00) <LLOQ(2.50) <LLOQ(1.00) 43.2 2.11 2.08 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 49.7 4.76 3.32 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 34.7 <LLOQ(1.92) <LLOQ(1.36) 2.34 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 29.1 1.95 2.19 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 41.3 2.80 3.90 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 42.1 2.96 2.31 2.50 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 32.0 3.03 1.89 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 18.5 <LLOQ(1.92) <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 20.2 2.64 <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 25.3 2.67 1.40 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 54.6 3.05 2.30 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 27.8 3.23 1.92 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 21.9 <LLOQ(1.92) <LLOQ(1.36) 1.72 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 55.2 3.06 2.16 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 37.5 2.78 <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 23.5 5.96 <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 35.7 2.08 <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 14.6 <LLOQ(1.92) <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 32.3 1.98 1.57 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 22.4 6.13 <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <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) o f 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 45 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Table 22. Serum Study Sample Concentrations (Continued) All concentrations are expressed as ppb. Sample PFOS No. PFOA* PFHS* PFOSAA* PFOSA* M556* M570* 41621 11.8 <LLOQ(1.92) <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 41623 20.3 2.39 <LLOQ(1.36) 1.60 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 41626 18.1 <LLOQ(1.92) <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 41633 20.0 <LLOQ(1.92) <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 41650 25.2 <LLOQ(1.92) <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(3.20) <LLOQ(1.90) 41665 25.2 2.42 <LLOQ(1.36) 1.99 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 41784 28.6 <LLOQ(1.92) 1.86 2.24 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 41859 18.4 <LLOQ(1.92) 1.63 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 42174 42.8 2.65 1.76 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 42524 32.1 5.78 1.50 2.79 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 42579 39.0 3.49 2.68 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 42621 40.5 2.32 2.16 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 42671 21.1 <LLOQ(1.92) <LLOQ(1.36) 2.95 <LLOQ(1.00) <LLOQ(2.50) 1.21 42818 19.5 3.06 <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 43095 34.5 2.13 2.95 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 43146 49.0 3.86 5.05 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 43246 50.3 2.76 2.72 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 43310 52.2 2.91 3.76 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 43484 22.4 <LLOQ(1.92) <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 43519 12.4 <LLOQ(1.92) <LLOQ(1.36) 6.73 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 43520 38.2 <LLOQ(1.92) 4.15 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 43542 12.5 2.39 2.58 1.96 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 43864 28.4 2.23 <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 43882 18.6 2.20 <LLOQ(1.36) 2.94 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 44145 34.4 <LLOQ(1.92) <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 44391 20.5 <LLOQ(1.92) <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 44448 38.3 2.16 2.60 2.42 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 45329 17.0 <LLOQ(1.92) <LLOQ(1.36) 4.31 <LLOQ(1.00) <LLOQ(2.50) 2.83 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 46 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Table 22. Serum Study Sample Concentrations (Continued) All concentrations are expressed as ppb. Sample No. 45330 PFOS PFOA* PFHS* 16.3 <LLOQ(l .92) <LLOQ(1.36) PFOSAA* 4.61 PFOSA* M556* <LLOQ(l .00) <LLOQ(2.50) M570* 2.67 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 47 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Table 23. Plasma Study Sample Concentrations All concentrations are expressed as ppb. Sample PFOS No. A-0150 22.9 A-0277 31.9 A-0334 61.8 A-0790 33.3 A-1093 16.7 A-1211 29.3 A-1212 41.3 A-1242 18.5 A-1332 13.1 A-1335 89.6 A-1880 41.2 A-1929 16.6 A-2007 27.0 A-2159 28.2 A-2527 20.1 A-2542 29.0 A-2759 36.9 A-2760 27.5 A-2919 29.6 A-3003 64.8 A-3133 42.9 A-3142 40.7 A-3151 10.5 A-3169 21.7 A-3234 17.3 A-3260 69.1 A-3297 33.4 A-3322 23.7 A-3327 41.9 PFOA 3.23 2.75 7.68 12.8 5.06 4.84 6.05 2.00 3.02 6.46 6.47 2.64 6.52 5.83 2.96 4.43 4.36 4.56 4.76 5.34 3.39 9.49 2.12 3.17 3.15 5.57 4.09 4.04 5.34 PFHS PFOSAA PFOSA M556 M570 <LLOQ(1.36) 3.29 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) <LLOQ(1.36) 5.18 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 7.91 3.78 <LLOQ(1.00) <LLOQ(2.50) 2.42 2.32 2.85 <LLOQ(1.00) <LLOQ(2.50) 1.80 2.31 2.87 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 1.91 2.50 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 2.71 2.59 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(l .00) <LLOQ(1.36) 3.08 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(l .00) <LLOQ(1.36) 1.77 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 6.88 6.07 <LLOQ(1.00) <LLOQ(2.50) 1.24 2.74 4.09 <LLOQ(1.00) 5.61 <LLOQ(1.00) <LLOQ(1.36) 3.15 <LLOQ(1.00) <LLOQ(2.50) 1.04 <LLOQ(1.36) 4.25 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) <LLOQ(1.36) 2.64 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 3.07 2.96 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 1.47 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 2.32 1.83 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) <LLOQ(1.36) 3.33 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) <LLOQ(1.36) 3.65 <LLOQ(1.00) <LLOQ(2.50) 2.09 2.55 2.44 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 2.30 3.88 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 1.42 8.80 <LLOQ(l .00) <LLOQ(2.50) <LLOQ(1.00) <LLOQ(1.36) 2.47 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 1.39 3.12 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) <LLOQ(1.36) 3.95 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 2.35 2.50 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 3.45 1.90 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) <LLOQ(1.36) 2.63 <LLOQ(l .00) <LLOQ(2.50) <LLOQ(1.00) 2.57 3.50 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(l .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. Page 48 Northwest Bioanalytical Study No. NWBSOO-091 Report No. NWBR01-065 Table 23. Plasma Study Sample Concentrations (Continued) All concentrations are expressed as ppb. Sample PFOS No. A-3354 23.3 A-3355 70.3 A-3380 29.0 A-3410 69.3 A-3439 61.7 A-3442 27.3 A-3447 40.7 A-3461 43.5 A-3532 56.8 A-3534 40.4 A-3600 38.6 A-3630 32.3 A-3631 17.9 A-3683 33.0 A-3796 19.2 A-3797 62.5 A-3834 48.6 A-3844 33.0 A-3869 58.0 A-3954 29.2 A-3955 22.9 A-3956 38.6 A-4060 45.0 A-4078 41.5 A-4097 19.6 A-4115 45.8 A-4173 60.5 A-4979 55.3 A-6362 30.9 PFOA 18.9 4.73 6.40 71.7 10.8 6.42 6.77 5.76 3.39 4.31 5.43 5.74 4.90 3.29 7.14 6.79 6.50 6.32 5.33 3.58 3.67 5.31 5.92 7.90 5.51 6.98 9.61 5.94 4.54 PFHS PFOSAA PFOSA M556 M570 <LLOQ(1.36) 6.10 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 3.09 5.72 <LLOQ(1.00) <LLOQ(2.50) 2.26 2.38 3.99 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 16.5 28.7 CLLOQ(I.OO) <LLOQ(2.50) 1.10 2.40 3.86 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 2.15 2.96 <LLOQ(1.00) <LLOQ(2.50) 1.07 2.23 3.41 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 11.4 3.83 cLLOQ(l.OO) <LLOQ(2.50) CLLOQ(I.OO) <LLOQ(1.36) 13.3 cLLOQ(l.OO) <LLOQ(2.50) <LLOQ(1.00) 4.55 5.22 <LLOQ(l .00) <LLOQ(2.50) 3.15 3.14 3.93 cLLOQ(l.OO) <LLOQ(2.50) <LLOQ(l .00) 1.79 2.93 cLLOQ(l.OO) <LLOQ(2.50) 2.14 <LLOQ(1.36) 2.60 <LLOQ(1.00) <LLOQ(2.50) 3.64 <LLOQ(1.36) 4.19 <LLOQ(l .00) <LLOQ(2.50) <LLOQ(1.00) <LLOQ(1.36) 2.27 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 4.65 <LLOQ(1.60) cLLOQ(l.OO) <LLOQ(2.50) 1.11 4.31 2.66 <LLOQ(l .00) <LLOQ(2.50) 1.63 1.77 2.87 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 2.59 3.42 <LLOQ(1.00) <LLOQ(2.50) 5.69 1.48 4.23 cLLOQ(l.OO) <LLOQ(2.50) <LLOQ(1.00) 1.94 2.02 <LLOQ(l .00) <LLOQ(2.50) <LLOQ(1.00) 2.54 3.60 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 14.1 7.14 <LLOQ(1.00) <LLOQ(2.50) 1.43 2.97 3.50 cLLOQ(l.OO) <LLOQ(2.50) 1.01 1.64 2.11 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 14.5 4.56 <LLOQ(l .00) <LLOQ(2.50) <LLOQ(1.00) 1.45 5.15. <LLOQ(1.00) 3.34 1.35 3.22 3.76 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 2.35 1.60 <LLOQ(1.00) <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. Page 49 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Table 23. Plasma Study Sample Concentrations (Continued) All concentrations are expressed as ppb. Sample PFOS No. A-6657 32.8 A-7535 45.5 A-7826 12.1 A-8008 28.4 A-8013 27.2 A-8028 53.6 A-8115 54.8 B-0585 35.2 B-1082 36.9 B-1112 20.8 B-1394 34.8 B-1739 12.2 B-1740 28.4 B-1783 50.1 B-2127 65.5 B-2170 33.4 B-2171 56.4 B-2199 40.3 B-2228 38.7 B-2584 58.6 B-2924 52.5 B-2938 45.8 B-3110 41.0 B-3114 31.8 B-3163 65.7 B-3169 59.1 B-3170 52.8 B-3171 50.0 B-3172 28.9 PFOA 6.70 5.59 2.05 5.39 5.58 5.63 5.99 3.51 5.62 2.95 5.23 3.03 4.51 6.40 6.14 4.39 7.78 5.33 5.55 6.42 10.9 7.32 5.26 3.99 8.58 5.85 5.77 5.19 5.50 PFHS PFOSAA PFOSA M556 M570 2.27 2.50 <LLOQ(1.00) <LLOQ(2.50) 1.29 2.84 3.64 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) <LLOQ(1.36) 4.30 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 3.15 1.96 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 3.11 2.36 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 1.44 4.40 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 1.60 4.91 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 1.55 1.79 <LLOQ(1.00) <LLOQ(2.50) 1.32 1.94 5.25 cLLOQ(l.OO) <LLOQ(2.50) 1.17 1.56 2.26 CLLOQ(I.OO) <LLOQ(2.50) 1.04 2.87 2.59 <LLOQ(1.00) <LLOQ(2.50) 1.77 7.17 4.05 <LLOQ(l .00) <LLOQ(2.50) <LLOQ(1.00) 6.51 1.97 cLLOQ(l.OO) <LLOQ(2.50) <LLOQ(1.00) 2.21 29.5 <LLOQ(1.00) 5.19 1.69 4.24 5.42 cLLOQ(l.OO) <LLOQ(2.50) 1.80 1.98 2.38 cLLOQ(l.OO) <LLOQ(2.50) <LLOQ(1.00) 4.22 3.34 <LLOQ(1.00) <LLOQ(2.50) 1.71 3.01 2.24 <LLOQ(1.00) <LLOQ(2.50) 2.86 1.45 5.42 <LLOQ(1.00) <LLOQ(2.50) 4.46 2.34 5.24 <LLOQ(1.00) <LLOQ(2.50) 1.11 2.56 28.0 <LLOQ(1.00) 6.63 <LLOQ(1.00) 3.97 4.06 <LLOQ(1.00) <LLOQ(2.50) 1.11 11.3 2.57 <LLOQ(1.00) <LLOQ(2.50) 1.84 1.41 3.90 <LLOQ(1.00) <LLOQ(2.50) 1.61 2.63 5.39 <LLOQ(l .00) <LLOQ(2.50) <LLOQ(1.00) 2.32 2.83 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 3.82 3.42 cLLOQ(l.OO) <LLOQ(2.50) cLLOQ(l.OO) 2.29 4.39 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) <LLOQ(1.36) 4.07 <LLOQ(1.00) <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. Page 50 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Table 23. Plasma Study Sample Concentrations (Continued) All concentrations are expressed as ppb. Sample PFOS No. PFOA PFHS PFOSAA PFOSA M556 M570 B-3195 24.2 4.56 1.87 2.90 <LLOQ(1.00) <LLOQ(2.50) 1.27 B-3197 48.2 6.97 3.63 5.91 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) B-3198 43.7 6.66 4.87 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) B-3206 6.50 <LLOQ(1.92) <LLOQ(1.36) 2.71 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) B-3211 39.7 5.41 4.37 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) 1.04 B-3212 38.6 6.46 2.96 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) 1.38 B-3225 40.7 6.76 15.2 2.30 <LLOQ(1.00) <LLOQ(2.50) 1.44 B-3241 20.7 3.05 <LLOQ(1.36) 2.08 <LLOQ(1.00) <LLOQ(2.50) 4.37 B-3242 22.7 2.66 <LLOQ(1.36) 2.45 CLLOQ(I.OO) <LLOQ(2.50) <LLOQ(1.00) B-3265 59.0 6.93 4.95 4.76 <LLOQ(1.00) <LLOQ(2.50) 1.25 B-3266 22.4 3.76 2.14 3.78 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) B-3276 32.9 5.69 2.18 3.65 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) B-3282 44.1 4.89 4.05 i 6.60 <LLOQ(1.00) <LLOQ(2.50) 5.46 B-3290 35.7 7.32 2.63 2.79 <LLOQ(1.00) <LLOQ(2.50) 1.33 B-3292 25.9 5.39 2.37 5.17 <LLOQ(1.00) <LLOQ(2.50) <LLOQ( 1.00) B-3402 32.5 6.35 3.89 4.50 <LLOQ(l .00) <LLOQ(2.50) <LLOQ(1.00) B-3563 33.6 5.01 1.57 2.78 <LLOQ(1.00) <LLOQ(2.50) 1.63 B-3575 13.6 4.49 1.57 2.20 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) B-3844 18.1 4.57 2.24 2.08 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) B-3852 36.1 6.43 3.43 4.31 <LLOQ(1.00) <LLOQ(2.50) 1.65 B-3948 41.0 5.54 2.64 5.16 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) B-3952 38.7 7.77 4.31 2.56 <LLOQ(1.00) <LLOQ(2.50) 1.01 B-3955 12.9 10.5 4.39 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) B-3960 18.1 2.91 3.09 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) B-3978 31.6 8.01 2.57 3.39 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) B-4054 11.6 3.32 1.60 4.71 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) B-4258 25.0 4.32 3.01 3.86 <LLOQ(1.00) <LLOQ(2.50) 1.51 B-4259 34.6 4.40 1.56 3.22 <LLOQ(1.00) <LLOQ(2.50) 1.57 B-4887 12.9 3.13 <LLOQ(1.36) 2.44 <LLOQ(1.00) <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. Page 51 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Table 23. Plasma Study Sample Concentrations (Continued) All concentrations are expressed as ppb. Sample PFOS No. B-5109 19.2 B-5434 29.5 B-5521 20.7 B-5548 37.5 B-5549 44.0 B-5966 32.1 B-6165 57.3 B-6201 16.5 B-6396 123 B-6397 23.0 B-6649 37.5 B-6883 34.9 B-7259 21.8 B-7512 24.8 B-7926 36.5 B-7936 36.3 B-7957 25.8 B-7962 24.2 F-0494 59.6 F-1385 25.3 F-2872 53.0 F-3028 18.2 F-3029 27.5 F-3118 41.4 F-3233 39.7 F-3367 51.1 F-3368 42.3 F-4089 51.0 F-4166 41.1 PFOA 5.51 6.21 2.77 6.21 6.05 8.38 12.5 4.14 11.0 4.43 22.0 5.95 6.52 6.37 14.8 13.5 4.53 3.89 19.0 4.43 7.27 6.00 3.33 5.73 5.69 6.57 5.86 9.63 6.71 PFHS PFOSAA PFOSA M556 M570 1.92 3.67 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 3.40 3.71 <LLOQ(1.00) <LLOQ(2.50) 3.74 9.89 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 4.18 3.09 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 3.34 6.65 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 8.98 2.47 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 30.3 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(3.20) <LLOQ(1.90) 2.47 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 5.86 127 <LLOQ(1.00) 4.32 1.27 1.67 3.00 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 65.4 2.58 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 9.16 4.98 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 2.10 2.36 <LLOQ(1.00) <LLOQ(2.50) 1.05 2.62 2.27 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 2.38 7.36 <LLOQ(1.00) <LLOQ(2.50) 2.64 1.95 7.61 <LLOQ(1.00) <LLOQ(2.50) 2.65 18.6 3.47 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 16.2 3.56 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 48.1 5.02 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) <LLOQ(1.36) 5.57 <LLOQ(1.00) <LLOQ(2.50) 1.34 3.23 3.30 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) <LLOQ(1.36) 3.00 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 1.83 2.25 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 2.15 12.4 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 13.3 2.42 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 3.22 40.9 <LLOQ(1.00) <LLOQ(2.50) 1.37 2.79 10.1 <LLOQ(1.00) <LLOQ(2.50) cLLOQ(l.OO) 11.3 8.91 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 3.13 4.04 <LLOQ(1.00) <LLOQ(2.50) 2.18 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. Page 52 Northwest Bioanalytical Study No. NWBSOO-091 Report No. NWBR01-065 Table 23. Plasma Study Sample Concentrations (Continued) All concentrations are expressed as ppb. Sample PFOS No. F-4441 25.7 F-4456 48.2 F-4516 40.9 F-4995 27.8 F-5050 38.0 F-5112 33.2 F-5156 21.7 F-5289 28.2 F-6133 31.7 F-6202 39.5 F-6610 25.8 F-6614 29.2 F-6715 39.1 F-6716 37.3 L-0840 63.4 L-0866 24.5 L-0868 48.2 L-0948 48.1 L-1484 10.3 L-3082 35.6 L-4078 58.1 L-4134 23.6 L-4535 16.6 L-4553 40.2 L-4637 61.1 L-4731 33.0 L-4845 27.5 L-5881 22.8 L-5945 18.4 PFOA 5.32 6.96 6.05 5.68 4.85 5.15 3.41 4.06 3.91 4.97 3.74 3.67 9.65 10.2 7.00 8.86 11.7 6.53 2.23 6.68 10.4 4.13 3.81 7.49 9.10 5.62 8.63 3.18 4.00 PFHS PFOSAA PFOSA M556 M570 3.08 <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) 1.39 2.59 3.16 CLLOQ(I.OO) <LLOQ(2.50) 1.33 1.56 2.69 CLLOQ(I.OO) <LLOQ(2.50) 1.20 5.17 2.00 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 6.65 3.07 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 2.46 3.42 <LLOQ(1.00) <LLOQ(2.50) 2.06 <LLOQ(1.36) <LLOQ(1.60) <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 1.82 1.82 CLLOQ(I.OO) <LLOQ(2.50) 1.06 3.82 4.62 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 1.78 12.4 <LLOQ(1.00) 2.70 <LLOQ(1.00) 3.02 9.23 <LLOQ(1.00) <LLOQ(2.50) 1.16 3.98 9.72 <LLOQ(1.00) <LLOQ(2.50) 1.22 5.50 2.06 <LLOQ(1.00) <LLOQ(2.50) 1.02 5.13 1.97 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(l .00) 3.42 11.5 <LLOQ(1.00) <LLOQ(2.50) 5.22 5.54 2.22 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 7.39 1.97 <LLOQ(1.00) <LLOQ(2.50) 1.89 4.15 3.68 <LLOQ(1.00) <LLOQ(2.50) 1.37 <LLOQ(1.36) 2.55 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 2.07 3.23 <LLOQ(1.00) <LLOQ(2.50) 1.34 2.27 4.86 <LLOQ(l .00) <LLOQ(2.50) <LLOQ(1.00) 1.64 3.10 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) <LLOQ(1.36) 1.85 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 6.57 2.79 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 2.28 86.0 <LLOQ(1.00) <LLOQ(2.50) 1.91 <LLOQ(1.36) 40.8 <LLOQ(1.00) <LLOQ(2.50) 1.14 40.3 3.42 cLLOQ(l.OO) <LLOQ(2.50) <LLOQ(1.00) <LLOQ(1.36) 3.39 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 1.59 <LLOQ(1.60) <LLOQ(1.00) <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. Page 53 Northwest Bioanalytical Study No. NWBSOO-091 Report No. NWBRO1-065 Table 23. Plasma Study Sample Concentrations (Continued) All concentrations are expressed as ppb. Sample PFOS No. L-7039 32.7 L-7225 32.1 L-7415 35.7 L-7472 30.0 L-7703 49.8 L-7709 43.7 L-7726 25.4 L-7728 31.9 M-0106 52.3 M-0160 25.9 M-0424 67.6 M-0474 37.1 M-2136 41.3 M-2213 42.4 M-2214 40.1 M-2228 32.2 M-2233 54.4 M-2247 33.3 M-3719 20.7 M-4002 78.8 M-4294 37.0 M-4726 26.1 M-4732 25.3 PFOA PFHS PFOSAA PFOSA M556 M570 5.60 3.25 2.48 <LLOQ(1.00) <LLOQ(2.50) 2.22 4.54 4.69 2.43 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 7.22 2.69 8.00 <LLOQ(l .00) 2.60 1.92 4.84 2.65 3.73 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(l .00) 6.26 1.89 4.28 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 5.87 1.84 3.66 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 4.41 . 6.44 5.88 <LLOQ(1.00) <LLOQ(2.50) 1.36 5.82 8.25 7.98 <LLOQ(1.00) <LLOQ(2.50) 1.65 5.91 3.55 4.40 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 5.86 1.61 7.19 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 8.85 6.08 3.66 <LLOQ(1.00) <LLOQ(2.50) 1.17 7.45 1.86 10.5 <LLOQ(1.00) <LLOQ(2.50) 2.44 6.34 1.51 4.90 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 4.89 2.03 2.89 <LLOQ(1.00) <LLOQ(2.50) 1.09 6.85 <LLOQ(1.36) 4.69 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 6.40 5.29 5.04 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 12.5 20.6 7.07 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 7.51 <LLOQ(1.36) 4.81 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 3.07 <LLOQ(1.36) 2.49 <LLOQ(1.00) <LLOQ(2.50) <LLOQ(1.00) 8.22 9.63 14.4 <LLOQ(1.00) <LLOQ(2.50) '2.36 5.99 8.73 2.90 <LLOQ(1.00) <LLOQ(2.50) 3.25 3.60 <LLOQ(1.36) 7.44 <LLOQ(1.00) <LLOQ(2.50) 2.18 3.23 <LLOQ(1.36) 7.00 <LLOQ(1.00) <LLOQ(2.50) 2.06 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. Page 54 Northwest Boanalytical Study No. NWBSOO-091 Report No. NWBRO1-065 Table 24. Repeat Analysis Table for PFOS in Human Serum Sample O riginal Cone. O riginal R un No. PPb N um ber 41650 21.1 10 Reason for Reassay 1 Reassay Cone, ppb 25.4, 24.9 Reassay Run N um ber 17, 17 R ep o rted Cone, ppb 25.2 Reason for Reported Cone. 1 REASONS FOR REASSAY: 1). Analytical reasons. REASONS FOR REPORTED CONC: 1). Mean o f reassay results. Original result invalid. Table 25. Repeat Analysis Table for PFO A in H um an Serum Sample No. O riginal Cone, ppb 41650 25.4 O rig in al Run N um ber 10 Reason for Reassay Reassay Cone, ppb 1 <LLOQ(1.92), <LLOQ(1.92) Reassay Run N um ber 17, 17 Reported Cone. ppb <LLOQ(1.92) Reason for R ep o rted Cone. 1 REASONS FOR REASSAY: 1). Analytical reasons. REASONS FOR REPORTED CONC: 1). Mean of reassay results. Original result greater than 15% deviation from reassay results. Page 55 Northwest Bioanalytical Study No. NWBSOO-091 Report No. NWBR01-065 Table 26. Repeat Analysis Table for PFHS in Human Serum Sample O riginal No. Cone, ppb 41650 29.2 O rig in al Run N um ber 10 Reason for Reassay Reassay Cone, ppb 1 <LLOQ(1.36), <LLOQ(1.36) Reassay Run N um ber 17, 17 Reported Cone, ppb <LLOQ(1.36) Reason for R ep o rted Cone. 1 REASONS FOR REASSAY: 1). Analytical reasons. REASONS FOR REPORTED CONC: 1). Mean of reassay results. Original result greater than 15% deviation from reassay results. Table 27. R epeat Analysis Table for PFOSAA in H um an Serum Sample O riginal No. Cone, ppb 41650 23.2 O rig in al Run N um ber 10 Reason for Reassay Reassay Cone, ppb 1 <LLOQ(1.60), <LLOQ(1.60) Reassay Run N um ber 17, 17 Reported Cone, ppb <LLOQ(1.60) Reason for R ep o rted Cone. 1 REASONS FOR REASSAY: 1). Analytical reasons. REASONS FOR REPORTED CONC: 1). Mean of reassay results. Original result greater than 15% deviation from reassay results. Page 56 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBR01-065 Table 28. Repeat Analysis Table for PFOSA in Human Serum Sample O riginal No. Cone, ppb 41650 22.0 O rig in al Run N um ber 10 Reason for Reassay Reassay Cone, ppb 1 <LLOQ(1.00), <LLOQ(1.00) Reassay Run N um ber 17, 17 Reported Cone, ppb <LLOQ(1.00) Reason for R ep o rted Cone. 1 REASONS FOR REASSAY: 1). Analytical reasons. REASONS FOR REPORTED CONC: 1). Mean of reassay results. Original result greater than 15% deviation from reassay results. Table 29. R epeat Analysis Table for M 556 in H um an Serum Sample O riginal No. Cone, ppb 41650 20.4 O rig in al Run N um ber 10 Reason for Reassay Reassay Cone, ppb 1 <LLOQ(3.20), <LLOQ(3.20) Reassay Run N um ber 18,18 Reported Cone, ppb <LLOQ(3.20) Reason for R ep o rted Cone. 1 REASONS FOR REASSAY: 1). Analytical reasons. REASONS FOR REPORTED CONC: 1). Mean of reassay results. Original result greater than 15% deviation from reassay results. Page 57 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Table 30. Repeat Analysis Table for M570 in Human Serum Sample O riginal No. Cone, ppb 41650 23.8 O riginal Run N um ber 10 Reason for Reassay Reassay Cone, ppb 1 <LLOQ(1.90), <LLOQ(1.90) Reassay Run Number 18, 18 Reported Cone, ppb <LLOQ(L90) Reason for R ep o rted Cone. 1 REASONS FOR REASSAY: 1). Analytical reasons. REASONS FOR REPORTED CONC: 1). Mean of reassay results. Original result greater than 15% deviation from reassay results. Page 58 Northwest Bioanalytical Study No. NWBSOO-091 Report No. NWBRO1-065 Figure 1. Representative Calibration Curve for PFOS Analytical R un 3 analyzed on 14-O ct-2000 C alibration Standards for PFO S (ppb) Regression M ethod * Q U A D R A TIC - W eighting Factor * 1/X**2 Quadratic Lim it - 2200 Instrument Response Figure 2. R epresentative C alibration C urve for PFO A Analytical R un 3 analyzed on 14-0ct-2000 Calibration Standards for PO A A (ppb) R eg ressio n M e th o d Q U A D R A T IC - W eig h tin g F a c to r * 1/X **2 Q uadratic L im it 571000 Instrument Response Page 59 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Figure 3. Representative Calibration Curve for PFHS Analytical R un 3 analyzed on 14-0ct-2000 Calibration Standards for PFH S (ppb) R e g re ssio n M e th o d =Q U A D R A T IC - W eig h tin g F a c to r * 1/X **2 Quadratic Lim it ] 600 Instrumoit R ponse Figure 4. Representative C alibration C urve for PFOSAA A nalytical R un 3 analyzed on 14-0ct-2000 C alibration Standards for PFO SA A (ppb) R eg ressio n M e th o d * Q U A D R A T IC - W eig h tin g F a c to r = 1 / X mm2 Quadratic Lim it = -4280 Instrument Response Page 60 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBR01-065 Figure 5. Representative Calibration Curve for PFOSA Analytical Run 3 analyzed on 14-0ct-2000 C alibration Standards for PFO SA (ppb) Regression M ethod = QUAD RA TIC - W eighting Factor * 1/X**2 Q uadratic L im it *=5900 Instrument Response Figure 6. R epresentative C alibration C urve for M 556 Analytical R un 3 analyzed on 14-0ct-2000 C alibration Standards for M 556 (ppb) R eg ressio n M eth o d " Q U A D R A T IC - W eighting F a c to r ,si 1/X **2 Quadratic Lim it = 108000 Instrument Response Page 61 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Figure 7. Representative Calibration Curve for M570 Analytical Run 3 analyzed on 14-0ct-2000 C alibration Standards for M 570 (ppb) Regression M ethod * Q U AD RA TIC - W eighting F actor = 1/X**2 Q uadratic Lim it * 3820 Page 62 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBR01-065 Figure 8. Human Plasma Blank for PFOS I PFOS Internal Standard: THPFOS Use Area Absolute Retention Time Expected RT 4.96 I Current Method Noise Thres. 5.0 5.0 Quant Thres. 0.2 0.2 Min. Width 33 Mult. Width 12 12 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 S009105Q11 5 011 S00091 CONTROL BLANK 1 Mon, Oct 16.2000 9:25 PM 0.98 in 1 period io d '1 1: 8.98 MRM, 571 scans 499.0->80.0 Area 8446 Heiaht 558 Start Time End Time Integration Width Retention Time Intearation Type HHEM 4.54 5.17 0.63 4.77 A -B B 98- p If,7C; mm 1g1g1g . 20 10 o+~* No Comment 1THPFOS use as Internal Standard Expected RT 4.32 I Current Method Noise Thres. 10.0 10.0 Quant Thres. 1.0 1.0 Min. Width 33 Mult. Width 55 oo Base. Width 100 RT Win. (secs) 20 20 Smooth 1 1 S009105011 5 011 S00091 CONTROL BLANK 1 No Comment Page 63 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBR01-065 Figure 9. Human Plasma Blank for PFOA IPOAA Internal Standard: THPFOS Use Area Absolute Retention Time Expected RT 4.41 Current Method Noise Three. 1.0 1.0 Quant Thres. 0.5 0.5 Min. Width 33 Mult. Width 88 Base. Width 100 100 RT Win. (sacs) 20 20 Smooth 1 1 S009105011 5 011 S00091 CONTROL BLANK 1 Mon, Oct 16. 2000 9:25 PM 8.98 in 1 period 1: 8.98 MRM, 571 scans 413.0->169.0 No Comment Area 758 Heiaht 73 Start Time End Time Intearation Width Retention Time Integration Type I t l+HHia 4.14 4.49 0.35 4.37 A -B B HU ITHPFOS use as Internal Standard Expected RT 4.32 1 Current Method Noise Thres. 10.0 10.0 Quant Thres. 1.0 1.0 Min. Width 33 Mult. Width 55 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 5 011 SQQ091 C O N T P m ni a kik 1 Kir r.nmmivil Page 64 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBR01-065 Figure 10. Human Plasma Blank for PFHS IPFHS Internai Standard: THPFOS Use Area Absolute Rtention Time Expected RT 3.62 I Current Method Noise Thres. 3.0 3.0 Quant Thres. 0.5 0.5 Min. Width 44 Mult. Width 10 10 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 s009105011 5 011 S00091 CONTROL_BLANK 1 Mon, Oct 16, 2000 9:25 PM 8.98 in 1 period m S mmmM 1: 6.98 MRM, 571 scans 399.0->80.0 Area 1008 Heiqht 125 Start Time End Time Intearation Width Retention Time Integration Type 3.64 3.96 0.32 3.73 A-BB Hitrattv:1S tps `V`-U" , f f :'J Mw-.5: *x h ' '' ?!\ ITHPFOS use as Internai Standard Expected RT 4.32 I Current Method Noise Thres. 10.0 10.0 Quant Thres. 1.0 1.0 Min. Width 33 Mult. Width 55 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 5 011 SOQ091 CONTROL BLANK 1 Mon, Oct 16, 2000 9:25 PM 8.98 in 1 period 4 s| 1: 8.98 MRM, 571 scans 427.0->407.0 Area 0 Height 0 Start Time End Time Intearation Width Retention Time mmIntearation Type 'US "Qi 0.00 0.00 0.00 s a 0.00 toi ;oi "9 S17 Page 65 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBR01-065 Figure 11. Human Plasma Blank for PFOSAA IPFOSAA Internal Standard: THPFOS Use Area I Current Method Noise Thres. 4.0 4.0 Quant Thres. 1.0 1.0 Min. Width 3 3 Absolute Retention Time Mult. Width 12 12 Expected RT 6.05 Base. Width 100 100 RTWm. (secs) 20 20 Smooth 1 1 S009105011 5 011 SOOO01 CONTROL_BLANK 1 No Comment - tvtwwttv: Mon. Oct 16, 2000 9:25 PM 8.96 in period * 'S ^ M * * - *S I S 1: 8.98 MRM. 571 scans 584,1->419.1 re. f ' - rr - l l *& * ' *>4': A*; `f t Area 2759 Height 340 Start Time End Time Integration WkJth Retentlon Time Integration Tvpe acaay 5.67 6.10 0.43 5.91 A -B B ii`-. =. J s.' [THPFOS use as internal Standard Expected RT 4.32 I Current Method Noise Thres. 10.0 10.0 Quant Thres. 1.0 1.0 Min. Width 3 3 M ult Width 55 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 S009105011 5 011 S00091 CONTROL BLANK 1 No Comment Page 66 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBR01-065 Figure 12. Human Plasma Blank for PFOSA IPFOSA ' Internal Standard: THPFOS U9e Area Absolute Retention Time Expected RT 5.59 ) Current Method Noise Thres. 3.0 3.0 Quant Thres. 0.1 0.1 Min. Width 55 Mult. Width 10 10 Base. Width 100 100 RT Win. (sec9) 20 20 Smooth 1 1 S009105011 5 011 S00091 CONTROL BLANK 1 Mon, Oct 16,2000 9:25 PM 8.96 in 1 period 1: 8.98 MRM, 571 scans 496.0->78.0 N< ** Area 965 Height 108 Start Time End Time Integration Width Retention Time integration Type B ilC C M 5.48 5.78 0.30 5.66 A -W ITHPFOS use as Internal Standard Expected RT 4.32 I Current Method Noise Thres. 10.0 10.0 Quant Thres. 1.0 1.0 Min. Width 33 Mult. Width 55 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 s009105011 5 011 S00091 CONTROL_BLANK 1 No Comment Page 67 Northwest Bioanalytical Study No. NWBSOO-091 Report No. NWBRO1-065 Figure 13. Human Plasma Blank for M556 IM556 internai Standard: THPFOS Use Area I Current Method Noise Thres. 1.0 1.0 Quant Thres. 0.1 0.1 Min. Width 33 Absolute Retention Time Mult. Width 16 18 Expected RT 5.36 Base. Width 200 200 RT Win. (secs) 20 20 Smooth 1 1 5 011 S0081 CONTROL BLANK 1 Mon. Oct 16. 2000 9:25 PM 8.98 in 1 period 1: 8.98 MRM, 571 scans i 556.0->498.0 ^> 8QJ Area 905 Heiaht 138 Start Time End Time Inteoration Width Retention Time Integration Type 5.23 5.52 0.28 5.39 A -B B 7a; : J l'i I '* * : h h c S bI HH m 20' IT* 10 0 w i t ITHPFOS use as Internal Standard Expected RT 4.32 Current Method Noise Thres. 10.0 10.0 Quant Thres. 1.0 1.0 Min. Width 33 Mult. Width 55 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 S09105011 5 011 S00091 CONTROL BLANK 1 No Comment mTimmae!- Page 68 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Figure 14. Human Plasma Blank for M570 [M57--------------------------------------1 >------- I Internal Standard: THPFOS Use Area Absolute Retention Time Expected RT 5.09 Current Method Noise Thres. 3.0 3,0 Quant Thres. 0.2 0.2 Min. Width 55 Mult. Width 15 15 Base. Width 100 100 RT Win, (secs) 20 20 Smooth 1 1 s009105011 5 011 S00091 CONROL_BU\NK 1 Mon, Oct 16. 2000 9:25 PM 8.98 in 1 period raa 1: 8.98 MRM, 571 scans ag- 570.0->419.1 80 Area 1408 Heiaht 202 Start Time End Time Inteoration Wldth Retention Time Integration Type 5.47 5.86 0.39 5.67 A * BB 10; 4 M'ji' 4t) 30*1 20 10 No C crrnent {THPFOS" use as Internal Standard Expected RT 4.32 Current Method Noise Thres. 10.0 10.0 Quant Thres. 1.0 1.0 Min. Width 3 3 Mult. Width 55 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 S091Q5011 5011 S00091 CONTROL BLANK 1 NcCom neni Page 69 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Figure 15. Human Plasma Blank with Internal Standard (QCO) for PFOS IPFOS Internal Standard: THPFOS Use Area I Current Method Noise Thres. 5.0 5.0 Quant Thres. 0.2 0.2 Min. Width 33 Absolute Retention Time Mult. Width 12 12 Expected RT 4.96 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 5 001 S00091 QC 0 1 It h p f o s use as Internal Standard Expected RT 4.32 I Current Method Noise Thres. 10.0 10.0 Quant Thres. 1.0 1.0 Min. Width 33 MuIL Width 55 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 S009105001 5 001 S0QQ91 QC 0 1 No Comment Page 70 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Figure 16. Human Plasma Blank with Internal Standard (QCO) for PFOA IPOAA ... I Current Method Internal Standard: THPFOS Use Area Noise Thres. Quant Thres. Min. Width 1.0 1.0 0.5 0.5 33 Absolute Retention Time Mult. Width 88 Expected RT 4.41 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 S009105001 5 001 S00091 QC 0 ' Mon. Oct 16. 2000 7:28 PM 8.98 in 1 period 1: 6.98 MRM, 571 scans 413.0->169.0 Area 1050 Height 104 Start Time End Time Intearation Width Retention Time Integration Type a n n a li 4.14 4.55 0.41 4.44 A-BB It h p f o s use as Internal Standard Expected RT 4.32 I Current Method Noise Thres. 10.0 10.0 Quant Thres. 1.0 1.0 Min. Width 33 Mult. Width 55 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 S009105001 5 001 S00091 QC 0 1 No Comment Page 71 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Figure 17. Human Plasma Blank with Internal Standard (QCO) for PFHS U-- ,1^ ------- ---- Internal Standard: THPFOS Use Area 1 Current Method Noise Thres. 3.0 3.0 Quant Thres. 0.5 0.5 Min. Width 4 4 Absolute Retention Time Mult. Width 10 10 Expected RT 3.62 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 S009105Q01 5 001 S00091QC 01 Mon. Oct 16. 2000 7:28 PM 8.98 in 1 period 1: 8.98 MRM, 571 scans 399.0->80.0 Area 879 Height 95 Start Time End Time tntearation Width Retention Time Integration Type H l+Jg M M 3.51 3.83 0.32 3.77 A - BB H ' N nt It h p f o s use as Internal Standard Expected RT 4.32 I Current Method Noise Thres. 10.0 10.0 Quant Thres. 1.0 1.0 Min. Width 33 Mult. Width 55 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 S009105001 5 001 S00091 QC 0 1 No Comment Page 72 Northwest Bioanalytical Study No. NWBS00-091 Report No. NWBRO1-065 Figure 18. Human Plasma Blank with Internal Standard (QCO) for PFOSAA IPF--OSAA- -------- Internal Standard: THPFOS Use Area 1\ Current Method Noise Thres. 4.0 4.0 Quant Thres. 1.0 1.0 Min. Width 33 Absolute Retention Time Mult Width 12 12 Expected RT 6.05 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 11 sQ09105001 a 001 S00091 QC 0 Mon, Oct 16. 2000 7:28 PM 8.98 in 1 period T. 8.98 MRM, 571 scans 584.1->419.1 Area 1124 Height 193 Start Time End Time integration Width Retention Time integration Type It llfr M M 5.72 6.02 0.30 5.89 A -B B IH B ITHPFOS use as Internal Standard Expected RT 4.32 I Current Method Noise Thres. 10.0 10.0 Quant Thres. 1.0 1.0 Min. Width 33 Mult. Width 55 Bass. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 SQ09106Q01 5 001 S00091 QC 0 1 Mon, Oct 16, 2000 7:28 PM 8.98 in 1 period 1: 8.98 MRM, 571 scans 427,0->407.0 Area 118018 Height 17045 Start Time End Time Integration Width Retention Time Integration Type It l+HHBI 4.19 4.57 0.38 4.38 A * BB EQ No Comment Page 73 Northwest Bioanalytical Study No. NWBSOO-091 Report No. NWBRO1-065 Figure 19. Human Plasma Blank with Internal Standard (QCO) for PFOSA Ip f o s a Internal Standard: THPFOS Use Area Absolute Retention Time I Current Method Noise Thres. 3.0 Quant Thres. 0.1 Min. Width Mult. Width _10_ 5 10 Expected RT 5.59 Base. Width 100 100 RT Win. (secs) 20 Smooth 1 s009105001 5 001 S0091 QC Mon, Oct 16. 2000 7:28 PM 8.98 in 1 period 1: 8.98 MRM, 571 scans 498.0->78.0 Area 0 Height 0 Start Time End Time Intearation Width Retention Time Integration Type 0.00 0.00 0.00 0.00 ITHPFOS use as Internal Standard Expected RT 4.32 I Current Method Noise Thres. 10.0 10.0 Quant Thres. 1.0 1.0 Min. Width 33 Mult. Width 55 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 S08105Q01 5 001 S00091 QCO 1 No Comment Page 74 Northwest Bioanalytical Study No. NWBSOO-091 Report No. NWBRO1-065 Figure 20. Human Plasma Blank with Internal Standard (QCO) for M556 |M556 Internal Standard: THPFOS Use Area I Current Method Noise Thres. 1.0 1.0 Quant Thres. 0.1 0.1 Min. Width 33 Absolute Retention Time Mult. Width 18 1B Expected RT 5.36 Base. Width 200 200 RT Win. (secs) 20 20 Smooth 1 1 SD091050Q1 5 001 SOOO1 QC 0 1 Mon. Oct 18. 20007:28 PM 8.98 in 1 period 1: 8.98 MRM, 571 scans 556.0->498.0 Area 0 Heiqht 0 Start Time End Time Inteoration Width Retention Time Inteoration Type EH9CZ 0.00 0.00 0.00 0.00 : 17 ITHPFOS use as Internal Standard Expected RT 4.32 I Current Method Noise Thres. 10.0 10.0 Quant Thres. 1.0 1.0 Min. Width 33 Mult. Width 55 Base. Width 100 1O0 RT Win. (secs) 20 20 Smooth 1 1 $009106001 5 001 S00091 QC 0 1 No Comment Page 75 Northwest Bioanalytical Study No. NWBSOO-091 Report No. NWBRO1-065 Figure 21. Human Plasma Blank with Internal Standard (QCO) for M570 IM570 Internal Standard: THPFOS Use Area I Current Method Noise Thres. 3.0 3.0 Quant Thres. 0.2 02 Min. Width 55 Absolute Retention Time Mult. Width 15 15 Expected RT 5.89 Base. Width 100 100 RT Win. {secs) 20 20 Smooth 1 1 S009105001 5 001 S000S1 QC Mon, Oct 18, 2000 7:28 PM 8.98 in 1 period 1: 6.98 MRM, 571 scans 570.0->419.1 kl" ----- * Area 0 Heiaht 0 Start Time End Time Intearation Width Retention Time Integration Type H H C 2.' 0.00 0.00 0.00 0.00 ITHPFOS use as Internal Standard Expected RT 4.32 I Current Method Noise Thres. 10.0 10.0 Quant Thres. 1.0 1.0 Min. Width 33 Mult. Width 55 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 s009105001 5 001 S00091 QC 0 1 No Comment Page 76 Northwest Bioanalytical Figure 22. Low Standard for PFOS Study No. NWBS00-091 Report No. NWBR01-065 Ip fo s Internal Standard: THPFOS Use Area Absolute Retention Time Expected RT 4.96 Current Method Noise Thres. 5.0 5.0 Quant Thres. 0.2 0.2 Min. Width 33 Mult Width 12 12 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 S009105002 5 002 S00091 Std 1 1 1 Mon. Oct 16. 2000 7:40 PM 8.98 in 1 period loe- 1: 8.98 MRM, 571 scans 499.0->80.0 ' 3i - BG* V i M ,1.' ; Area 11564 Height 741 Start Time End Time Integration Width Retention Time integration Type 4.52 5.15 0.63 4.98 A-BB i - 1. 3 ' IHHi - I '- 't f i ^ ' ' W fcV " * - ' *. , D a Q f f iW H Q il y s 3 | K g * >? . \ t J. 4 * ~r :e ! X- ^ Vi IB ; -T.IL. H ' ' t i o< 101 19 < ITHPFOS use as Internal Standard Expected RT 4.32 I Current Method Noise Thres. 10.0 10.0 Quant Thres. 1.0 1.0 Min. Width 33 MuiL Width 55 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 S009105002 5 002 S00091 Stell 1 1 No Comment Mon. Oct 16. 2000 7:40 PM . 4=- r . ' - , l-iU"nW-t* ;-?2 - ^ 3 i 8.98 in 1 period 1: 8.98 MRM, 571 scans . ' >* I * i.'li . 427.0-M07.0 Area 152577 - fe i Wk ' !T t ' il ' A | | | | | Height 22509 Start Time End Time Integration Width Retention Time Integration Type 4.19 4.60 0.41 4.38 A-BB . t **, ' I '* ' >m -/ r* ; t -i" ; - ; V H 2 !n .v . * m-US. %- h *:' 4 iti+ H H a S. : - ? lllil mi 20) aui 4eDu1i > .louij "SoWtarifi 1 9 .317* 7 33 Page 77 Northwest Bioanalytical Figure 23. Low Standard for PFOA IPOAA Internal Standard: THPFOS Use Area Absolute Retention Time Expected RT 4.41 I Current Method Noise Thres. 1.0 1.0 Quant Thres. 0.5 0.5 Min. Width 33 M ult Width 88 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 S009106002 5 002 S00091 Std1 1 Mon. Oct 16. 2000 7:40 PM 8.98 in 1 period 1: 8.98 MRM. 571 scans 413.0->169.0 Area 3070 Height 329 Start Time End Time Integration Width Retention Time Integration Type . 4.07 4.55 0.49 4.40 A -B B H ^ --------- - HWmBmBBB Study No. NWBS00-091 Report No. NWBR01-065 1THPFOS use as Internal Standard Expected RT 4.32 Current Method Noise Thres. 10.0 10.0 Quant Thres. 1.0 1.0 Min. Width 33 Mult. Width 55 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 S009105002 5 002 S00091 Std1 1 1 Mon. Oct 16. 2000 7:40 PM 8.98 in 1 period 1: 8.98 MRM. 571 scans 427.0->407.0 Area 152577 Height 22509 Start Time End Time Integration Width Retention Time Integration Type It l+HHl 4.19 4.60 0.41 4.38 A-BB No Comment Page 78 Northwest Bioanalytical Figure 24. Low Standard for PFHS IPFHS Internal Standard: THPFOS Use Area Absolute Retention Time Expected RT 3.62 I Current Method Noise Thres. 3.0 3.0 Quant Thres. 0.5 0.5 Min. Width 44 Mult. Width 10 10 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 S009105002 5 002 S00091 Std1 t 1 Study No. NWBS00-091 Report No. NWBRO1-065 ITHPFOS use as Internal Standard Expected RT 4.32 I Current Method Noise Thres. 10.0 10.0 Quant Thres. 1.0 1.0 Min. Width 33 M ult Width 55 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 S009105002 5 002 S00091 Std1 Mon, Oct 16,2000 7:40 PM 8.98 in 1 period 1: 8.90 MRM, 571 scans 427.0->407.0 Area 152577 Heiaht 22509 Start Time End Time Intecration Width Retention Time Inteqration Tvoe 4.19 4.60 0.41 4.38 A-BB jfcBfutnJE * - Page 79 Northwest Bioanalytical Figure 25. Low Standard for PFOSAA Study No. NWBSOO-091 Report No. NWBRO1-065 IPFOSAA Internal Standard: THPFOS Use Area Absolute Retention Time Expected RT 6.05 Current Method Noise Thres. 4.0 4.0 Quant Thres. 1.0 1.0 Min. Width 3 3 Mutt. Width 12 12 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 5 002S00091 Std1 Mon. Oct 16. 2000 7:40 PM 8.98 in 1 period 1 1: 8.98 MRM, 571 scans -t mte-nsttg "36 # 375 584.1->419.1 0 Area 5691 Height 725 Start Time End Time Integration Width Retention Time Integration? TTyyppe6 70 5.61 60 6.11 0.50 Z f 5.91 aA-- Bb bB _ | ^ i m k[M HiUi Bl lOE1 ' ? % 'i,r>r*p Jt!e 'aa| V H .;i ' ; ' .Is n:' iS ' ' : ' j -,t ' L .>* '1 %> l'l? i j i y* * ;i9y. &. ft ITHPFOS use as Internal Standard Expected RT 4.32 I Current Method Noise Thres. 10.0 10.0 Quant Thres. 1.0 1.0 Min. Width 33 Mult Width 55 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 s009105002 5 002 S00091 Std1 1 1 No Comment Page 80 Northwest Bioanalytical Figure 26. Low Standard for PFOSA Study No. NWBS00-091 Report No. NWBRO1-065 fPFOSA Internai Standard: THPFOS Use Area Absolute Retention Time Expected RT 5.59 1 Current Method Noise Thres. 3.0 3.0 Quant Thres. 0.1 0.1 Min. Width 55 Mult. Width 10 10 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 5 002 S00091 Std1 1 * ITHPFQS use as Internal Standard Expected RT 4.32 | Current Method Noise Thres. 10.0 10.0 Quant Thres. 1.0 1.0 Min. Width 33 Mult. Width 55 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 s009105002 5 002 S00091 Std1 1 1 Mon. Oct 16, 2000 7:40 PM 6.98 in 1 period 1: 8.98 MRM, 571 scans 427.0->407.0 KJV* 30; ao Area 152577 Heiaht 22509 Start Time End Time Inteqration Width Retention Time Integration Tvoe ItliHHH 4.19 4.60 0.41 4.38 A-BB 13 'TO: ,, 60 : 50- ; 40- ' to i 201 301 401 101 S u r ----1 3 1 7 * 7 4 6 33 7 93 TWjfc Page 81 Northwest Bioanalytical Figure 27. Low Standard for M556 Study No. NWBS00-091 Report No. NWBRO1-065 IM556 Internal Standard: THPFOS Use Area Absolute Retention Time Expected RT 5.36 I Current Method Noise Thres. 1.0 1.0 Quant Thres. 0.1 0.1 Min. Width 33 Mult Width 18 18 Base. Width 200 200 RT Win. (secs) 20 20 Smooth 1 1 s009105003 5 003 S00091 Std2 1 1 Mon. Oct 16. 2000 7:52 PM 8.98 in 1 period tit (flirty ; 90S tpi 3*4 1: 8.98 MRM, 571 scans 556.0->498.0 Sii Area 6787 7G Heiaht 907 Start Time 5.12 AO-- End Time Inteoration Width Retention Time Integration Type 5.58 0.46 5.42 A-BB r , *' ; . { . i--? , ;7 ;;$ ; % UuQUIfw ITHPFS use as Internal Standard Expected RT 4.32 I Current Method Noise Thres. 10.0 10.0 Quant Thres. 1.0 1.0 Min. Width 3 3 Mult. Width 55 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 S009105003 5 003 S00091 Std2 1 t No Comment Page 82 Northwest Bioanalytical Figure 28. Low Standard for M570 Study No. NWBS00-091 Report No. NWBRO1-065 Mayo Internal Standard: THPFOS Use Area I Current Method Noise Thres. 3.0 3.0 Quant Thres. 0.2 0.2 Min. Width 55 Absolute Retention Time M ult Width 15 15 Expected RT 5.89 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 S009105002 5 002 S00091 Sld1 Mon, Oct 16. 2000 7:40 PM 8.98 in 1 period 1: 8.98 MRM. 571 scans 570.0->419.1 Area 7142 Height 1105 Start Time End Time intearation Width Retention Time Integration Type 5.44 5.88 0.44 5.69 A-BB mm Intarsi** n 09&PS n 1A i ITHPFOS use as Internal Standard Expected RT 4.32 Current Method Noise Thres. 10.0 10.0 Quant Thres. 1.0 1.0 Min. Width 3 3 Mult. Width 55 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 S0091050Q2 5 002 S00091 SUJ1 1 1 No Comment Page 83 Northwest Bioanalytical Figure 29. High Standard for PFOS Study No. NWBS00-O91 Report No. NWBR01-065 IPFOS Internal Standard: THPFOS Use Area Absolute Retention Time Expected RT 4.96 I Current Method Noise Thres. 5.0 5.0 Quant Thres. 0.2 0.2 Min. Width 33 Mult. Width 12 12 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 sD09105010 6 010 S00091 Sld9 ' Mon. Oct 16. 2000 9:13 PM 6.98 in 1 period 1: 8.98 MRM, 571 scans 499.0*>80.0 Area 1221600 Height 120590 Start Time End Urne Integration Width Retention Time Integration Type css 4.41 5.47 1.06 4.98 A-BV *4 iSdS?.! cpj --HEBBHEI --IIBIIBBIIBIBI v r ,- ;lt. a , 4 :1 ITHPFOS use as Internal Standard Expected RT 4.32 I Current Method Noise Three. 10.0 10.0 Quant Thres. 1.0 1.0 Min. Width 3 3 Mult Width 55 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 5 010 S00091 Std9 1 1 Page 84 Northwest Bioanalytical Figure 30. High Standard for PFOA Study No. NWBS00-091 Report No. NWBRO1-065 [POAA Internal Standard: THPFOS Use Area Absolute Retention Time Expected RT 4.41 I Current Method Noise Thres. 1.0 1.0 Quant Thres. 0.5 0.5 Min. Width 3 3 Mult. Width 88 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 SQ910501Q 5 010 SQ091 SUJ9 J - Mon. Oct 16. 2000 9:13 PM 6.98 in 1 period F 1: 8.98 MRM, 571 scans 413.0->169.0 Area 951504 Heiqht 100477 StartTime End Time Intearation Width Rtention Time Intearation Type 3.94 4.82 0.88 4.43 A-BV ITHPFOS use as internal Standard Expected RT 4.32 Current Method Noise Thres. 10.0 10.0 Quant Thres. 1.0 1.0 Min. Width 33 Mult. Width 55 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 SQ09105C10 5 010 S00091 Std9 1 1 Mon. Oct 16. 2000 9:13 PM 8.98 in 1 period No Comment W r270 J InUnrlty 286S ef - . - - | | 1 | | *| 4 1: 8.98 MRM, 571 scans 427.0->407.0 W ilB lll ,t- illilli M i-g - v i Area 141962 Heiaht 20849 StartTime End Time 4.21 4.59 * laggsis? 10 - t ^ i r % fi ` f ? r j V |V ' iillsi iii * *liill -s g -- > ` - l : i"'. Intearation Width Retention Time Intearation Type im N n a 0.38 4.38 m ffl m i !l i 0 *f . V /.. l s i - J ? S : S A * %: J r | > g f f . \ * - i "' < " i XBp r* MM te * m * % . * II1 iii ' K.' ,, jjpjBjs ili#! H i flrjf HM Lfr ` .--i VQ! 201 T O ' *401 SEM *' 1 S9 3 17 4 3 4 -6 3 3 , fe Page 85 Northwest Bioanalytical Figure 31. High Standard for PFHS Study No. NWBSOO-091 Report No. NWBRO1-065 IPFHS Internal Standard: THPFOS Use Area Absolute Retention Time Expected RT 3.62 I Current Method Noise Thres. 3.0 3.0 Quant Thres. 0.5 0.5 Min. Width 4 4 M ult Width 10 10 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 SC915010 5 010 S0S1 Sld9 1 Mon. Oct 16. 2000 9:13 PM 8.98 in 1 period iOO 1: 8.98 MRM, 571 scans SC< 399.0->80.0 80 Area 1677751 Height 191933 Start Time End Time Integration Width Retention Time Integration Type DESK 3.28 4.29 1.01 3.75 A-BV 70. AO H i 1 2 2 t-tfru ttj 19l95B<pi llililliilliill jjgjPflj | ji| * iISilIIlill*lilll \ iMH ; *. m S B S B u B m ; . J t-f' ^ 1 - : ITHPFOS use as Internal Standard Expected RT 4.32 I Current Method Noise Thres. 10.0 10.0 Quant Thres. 1.0 1.0 Min. Width 3 3 Mult. Width 55 Base. Width 100 100 RT Win. (sees) 20 20 Smooth 1 1 S009105010 5 010 S00091 Std9 1 1 Mon. Oct 16. 2000 9:13 PM 8.96 in 1 period - , .4 int*n*ity;58&2p, 1: 8.98 MRM, 571 scans 90- ' 427.0*>4Q7.0 Area 141962 Heiaht 20849 Start Time End Time Integration Width Retention Time Inteoration Tvoe i+r*hnM 4.21 4.59 0.38 4.38 A-BB 80 ! !.4' ' .50m 40 !S8p|p8 23 30 MBB ' *i. - f - MV ] ,6 S ..-ji ?B -al i :? --I mM wwI 10 Wi :Fs iffll' * soi 3 " ? 4.74 <04 S32 7 90 SrilaSn-sI-- Page 86 Northwest Bioanalytical Figure 32. High Standard for PFOSAA Study No. NWBS00-091 Report No. NWBRO1-065 ItP-FO-- SAA Internai Standard: THPFOS Use Area Absolute Retention Time Expected RT 6.05 I Current Method Noise Thres. 4.0 4.0 Quant Thres. 1.0 1.0 Min. Width 33 Mult. Width 12 12 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 s009105010 5 010 S00Q1 Std9 ` Mon. Oct 16. 2000 9:13 PM 8.98 in 1 period 1: 8.98 MRM, 571 scans 584.1->419.1 Area 1725312 Height 222064 Start Time End Time Integration Width Retention Time integration Type h h eoe 5.44 6.60 1.17 5.91 A-BV ||g -IB ` m tm ltg 22208ST.W Jr5 Y": v 'V , ---j "ii' A * - iM mm T gni ITHPFQS use as Internai Stendard Expected RT 4.32 Current Method Noise Thres. 10.0 10.0 Quant Thres. 1.0 1.0 Min. Width 33 Mult. Width 55 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 s009105010 5 010 S00091 Std9 1 1 Mon, Oct 16.2000 9:13 PM 8.98 in 1 period 1: 8.98 MRM, 571 scans 427.0->407.0 Area 141962 Height 20849 Start Time End Time integration Width Retention Time Integration Type 4.21 4.59 0.38 4.38 A-BB No Comment Page 87 Northwest Bioanalytical Figure 33. High Standard for PFOSA Study No. NWBS00-091 Report No. NWBR01-065 IPFOSA Internai Standard: THPFOS Use Area Absolute Retention Time I Current Method Noise Thres. 3.0 Quant Thres. 0.1 Min. Width 5 Mutt. Width 10 10 Expected RT 5.59 Base. Width 100 100 RT Win. (secs) 20 Smooth 1 s009105010 5 010 S00091 Std9 1 1 Mon, Oct 16, 2000 9:13 PM 8.98 in 1 period 1: 8.98 MRM, 571 scans 498.0->78.0 Area 3016013 Height 331247 Start Time End Time Integration Width Retention Time Integration Type lU 5 M 5.20 6.87 1.67 5.67 A-BB ITHPFQS use as Internal Standard Expected RT 4.32 I Current Method Noise Three. 10.0 10.0 Quant Thres. 1.0 1.0 Min. Width 33 M ult Width 55 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 11 S009105010 5 010 S00091 Std9 1 1 No Comment Page 88 Northwest Bioanalytical Figure 34. High Standard for M556 IM556 Internal Standard: THPFOS Use Area Absolute Retention Time Expected RT 5.36 Current Method Noise Thres. 1.0 1.0 Quant Thres. 0.1 0.1 Min. Width 33 Mult. Width 18 18 Base. Width 200 200 RT Win. (secs) 20 20 Smooth 1 1 5 010 $00081 Sld9 Mon, Oct 16. 2000 S:13PM 8.98 in 1 period 1 : 8.98 MRM. 571 scans 556.0->498.0 Area 1328677 Height 165468 Start Time End Time Integration Width Retention Time Integration Type 4.89 6.76 1.88 5.41 A - BB Study No. NWBS00-091 Report No. NWBRO1-065 ITHPFOS use as Internal Standard Expected RT 4.32 Current Method Noise Thres. 10.0 10.0 Quant Thres. 1.0 1.0 Min. Width 33 Mult. Width 55 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 S009105010 5 01 S00091 Sld9 1 1 No Comment Page 89 Northwest Bioanalytical Figure 35. High Standard for M570 Study No. NWBS00-091 Report No. NWBR01-065 IM570 Internal Standard: THPFOS Use Area Absolute Retention Time Expected RT 5.89 I Current Method Noise Thres. 3.0 3.0 Quant Thres. 0.2 0.2 Min. Width 55 Mult. Width 15 15 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 5 010 SQ001 SUJ9 ' ' Mon. Oct 16. 2000 9:13 PM 8.98 in 1 period 1: 8.98 MRM, 571 scans 570.O->419.1 Area 2436405 Heiqht 326724 Start Time End Time Intearation Width Retention Time Inteqration Type 5.33 6.38 1.06 5.70 A-BV . 732Pi * 'y i fj.- ! $ s< . - t IJ - i; - r * ', <'*V:? % V it ITHPFOS use as Internal Standard Expected RT 4.32 I Current Method Noise Thres. 10.0 10.0 Quant Thres. 1.0 1.0 Min. Width 33 Mult. Width 55 Base. Width 100 100 RT Win. (secs) 20 20 Smooth 1 1 S009105010 5 010 S00091 Std9 1 1 Mon. Oct 16, 2000 9:13 PM 8.98 in 1 period 1: 8.98 MRM. 571 scans 427.0->407.0 Area 141962 Heioht 20849 Start Time End Time Intearation Width Retention Time Intearation Type 4.21 4.59 0.38 4.38 A-BB No Comment Page 90