Document X7aDErzB8YRpOGMrvRO5J7kyG
Northwest Bioanalytical
A R ASS - I A I 3
Study No. NWBSOO-145 Report No. NWBRO1-155
Quantitative Determination of PFOS and Related Compounds in Human Serum by LC/MS/MS
Sample Analysis Report for Protocol EPI-0016
2833FEB 19 AH Ih 08
Northwest Bioanalytical (NWB) A Division of NW T Inc. 1121 E ast 3900 South Salt Lake City, UT 84124
PREPARED FOR:
3M Company Corporate Occupational Medicine, Medical Department 220-3W-05 St. Paul, MN 55144
AUTHOR:
Connie O. Sakashita, B.S., NWB Project Manager
APPROVED FOR RELEASE BY:
M .^ Reoger L. Foltz, Ph.D.,
B Technical Director
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DATE: / - /c - 2-
DATE:
l-lo-O
MO 000599
Northwest Bioanalytcal
Study No. NWBSOO-145 Report No. NWBROl-155
QUALITY ASSURANCE STATEMENT
LABORATORY:
Northwest Bioanalytical (NWB) A Division of NWT Inc. 1121 East 3900 South Salt Lake City, UT 84124
SPONSOR:
3M Company Corporate Occupational Medicine, Medical Department 220-3W-05 St. Paul, MN 55144
COMPOUND(S):
PFOS and Related Compounds
NWB STUDY NUMBER:
NWBSOO-145
SPONSOR STUDY NUMBER: EPI-0016
NWB STUDY TITLE:
Quantitative Determination of PFOS and Related Compounds in Human Serum by LC/MS/MS
The clinical study described in this report is not included within the definition of a GLP regulated nonclinical study. However, Northwest Bioanalytical conducts all studies within the guidelines of the U.S. FDA Good Laboratory Practice Regulations for Nonclinical Laboratory Studies (Title 21 CFR Part 58), the OECD Principles of Good Laboratory Practice and the Japanese MHW Good Laboratory Practice Standard Ordinance for Nonclinical Laboratory Studies on the Safety of Drugs (Ordinance No. 21, PAB Notification No. 424.) The following inspections were performed by the NWB QAU per SOP.
Inspection and Reporting Statement.
Date of Inspection
Phase of Study
Date Report Sent to
Date Report Sent to
NWB Project Manager NWB Management*
22-23 Oct 2001 22-23 Oct 2001 24 Oct 2001 15-16 Nov 2001 07-09 Jan 2002 10 Jan 2002
Standards Preparation QC Preparation Analytical Plan Sample Analysis Report Draft/Raw Data Final Report
23 Oct 2001 23 Oct 2001 24 Oct 2001 16 Nov 2001 09 Jan 2002 10 Jan 2002
31 Oct 2001 31 Oct 2001 31 Oct 2001 30 Nov 2001 31 Jan 2002 31 Jan 2002
"Reports to NWB Management are issued monthly.
As can reasonably be established, the methods and procedures described and the results incorporated into this final report accurately reflect the raw data.
NWB Quality Assurance Compliance Auditor
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Date
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Study No. NWBS00-145 Report No. NWBR01-155
COMPLIANCE STATEMENT
The clinical study described in this report is not included within the definition o f a GLP regulated nonclinical study. However, to the best o f our knowledge, this project was conducted in accordance with the guidelines of the U.S. FDA Good Laboratory Practice Regulations for Nonclinical Laboratory Studies (Title 21 CFR Part 58) and according to the methods and procedures described within this report. In addition, the study followed the guidelines o f the OECD Principles of Good Laboratory Practice and the Japanese MHW Good Laboratory Practice Standard Ordinance for Nonclinical Laboratory Studies on the Safety o f Drugs (Ordinance No. 21, PAB Notification No. 424.) Any known incidents that may have affected the quality or integrity o f the project or reported data is included in this report. This report represents an accurate record o f the taw data.
Connie O. Sakashita, B.S. NWB Project Manager
/-/a - L . Date
/ --/ -- "2__ Date
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TABLE OF CONTENTS
Study No. NWBSOO-145 Report No. NWBRO1-155
SIGNATURE PAGE....................................................................................................................................1 QUALITY ASSURANCE STA TEM EN T............................................................................................ 2 COMPLIANCE STATEM ENT................................................................................................................ 3 TABLE OF CONTENTS..........................................................................................................................4 LIST OF TA B LES.......................................................................................................................................4 LIST OF FIGURES......................................................................................................................................5 1. INTRODUCTION............................................................................................................................... 8 2. METHODOLOGY..............................................................................................................................9 3. SAMPLE A N A LY SIS..................................................................................................................... 12 4. RESULTS AND DISCU SSIO N .....................................................................................................13 5. R EFEREN CES..................................................................................................................................14 6. DATA RETEN TIO N ........................................................................................................................15
LIST OF TABLES Table 1. Calibration Curve Summary for PFO S.................................................................................. 16 Table 2. Calibration Curve Summary for P F O A ................................................................................ 17 Table 3. Calibration Curve Summary for PFH S.................................................................................18 Table 4. Calibration Curve Summary for PFO SA A ...........................................................................19 Table 5. Calibration Curve Summary for PFO SA ...............................................................................20 Table 6. Calibration Curve Summary for M 556................................................................................. 21 Table 7. Calibration Curve Summary for M 570..................................................................................22 Table 8. Back-Calculated Concentrations o f Calibration Standards for P F O S ............................. 23
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Table 9. Back-Calculated Concentrations o f Calibration Standards for PFOA..............................24
Table 10. Back-Calculated Concentrations o f Calibration Standards for P F H S ...........................25
Table 11. Back-Calculated Concentrations o f Calibration Standards for PFOSAA..................... 26
Table 12. Back-Calculated Concentrations o f Calibration Standards for PFOSA........................ 27
Table 13. Back-Calculated Concentrations o f Calibration Standards for M 5 5 6 ...........................28
Table 14. Back-Calculated Concentrations o f Calibration Standards for M 5 7 0 ...........................29
Table 15. Analytical QC Summary for PFO S...................................................................................... 30
Table 16. Analytical QC Summary for PFO A ......................................................
31
Table 17. Analytical QC Summary for PFHS...................................................................................... 32
Table 18. Analytical QC Summary for PFO SA A ................................................................................33
Table 19. Analytical QC Summary for PFO SA .....................
34
Table 20. Analytical QC Summary for M 556...................................................................................... 35 )
Table 21. Analytical QC Summary for M 570...................................................................................... 36
Table 22. Serum Study Sample Concentrations........................................................ ..........................37
LIST OF FIGURES
Figure 1. Representative Calibration Curve for P F O S .......................................................................45
Figure 2. Representative Calibration Curve for PFO A ....................
45
Figure 3. Representative Calibration Curve for P F H S ....................................................................... 46
Figure 4. Representative Calibration Curve for PFOSAA................................................................. 46
Figure 5. Representative Calibration Curve for PFOSA.....................................................................47
Figure 6. Representative Calibration Curve for M556 .......................................................................47
Figure 7. Representative Calibration Curve for M 5 7 0 .......................................................................48
j Figure 8. Human Plasma Blank for PFO S............................................................................................49
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Figure 9. Human Plasma Blank for P F O A .......................................................................................... 50
Figure 10. Human Plasma Blank for PFH S......................................................................................... 5 1
Figure 11. Human Plasma Blank for PFO SA A ......................................................... .........................52
Figure 12. Human Plasma Blank for PFO SA ......................................................................................53
Figure 13. Human Plasma Blank for M 556......................................................................................... 54
Figure 14. Human Plasma Blank for M 570......................................................................................... 55
Figure 15. Human Plasma Blank with Internal Standard (QCO) for PF O S .....................................56
Figure 16. Human Plasma Blank with Internal Standard (QCO) for PFOA.....................................57
Figure 17. Human Plasma Blank with Internal Standard (QCO) for PF H S .....................................58
Figure 18. Human Plasma Blank with Internal Standard (QCO) for PFOSAA............................... 59
Figure 19. Human Plasma Blank with Internal Standard (QCO) for PF O SA ................................. 60
Figure 20. Human Plasma Blank with Internal Standard (QCO) for M 556.....................................61
Figure 21. Human Plasma Blank with Internal Standard (QCO) for M 5 7 0 .....................................62
Figure 22. Low Standard for PFOS........................................................................................................63
Figure 23. Low Standard for PFO A .......................................................................................................64
Figure 24. Low Standard for PFHS........................................................................................................65
Figure 25. Low Standard for PFOSAA................................................................................................. 66
Figure 26. Low Standard for PFOSA.................................................................................................... 67
Figure 27. Low Standard for M 556........................................................................................................68
Figure 28. Low Standard for M 570................................................................................................ 69
Figure 29. High Standard for PFO S.......................................................................................................70
Figure 30. High Standard for PF O A ......................................................................................................71
Figure 31. High Standard for PFH S.......................................................................................................72
Figure 32. High Standard for PFO SA A ................................................................................................ 73
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Figure 33. High Standard for PFOSA.................................................................................................. 74
Figure 34. High Standard for M 556......................................................................................................75
Figure 35. High Standard for M 570......................................................................................................76
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Study No. NWBS00-145 Report No. NWBROl-155
Quantitative Determination of PFOS and Related Compounds in Human Serum by LC/MS/MS
Sample Analysis Report for Protocol EPI-0016
1. IN T R O D U C T IO N
This report summarizes the analytical results from the quantitation o f PFOS, PFOA, PFHS, PFOSAA, PFOSA, M556, and M570 in human serum samples for 3M Company (3M) in support o f Protocol EPI-0016 [5.1]. The LC/MS/MS method for the analytes has a targeted calibration curve range o f 1.00 to 500 ppb (except for M556 which has a LLOQ o f 2.50 ppb). The actual calibration curve range for each analyte varies based upon the persistent levels o f the analyte in the matrix. For this study, the assay exhibited the following LLOQ and ULOQ values:
A n aly te PFOS PFOA PFHS
PFOSAA PFOSA M556 M570
LLOQ 3.37 ppb 1.44 ppb 1.36 ppb 1.50 ppb 1.00 ppb 2.50 ppb 1.00 ppb
ULOQ 413 ppb 481 ppb 522 ppb 501 ppb 500 ppb 500 ppb 500 ppb
The sponsor was 3M Company (Corporate Occupational Medicine, Medical Department, 220-3W-05, St. Paul, MN 55144), and the Study Director was Jeffrey H. Mandel, M.D., M.P.H. The Study Contact at 3M was Jean M. Burris. James K. Lundberg was the technical contact at 3M. The following is a list of NWB supervisory personnel involved in the completion o f this work: Connie O. Sakashita, B.S. (NWB Project Manager); Patrick Bennett, M.S., M.B.A. (NWB Laboratory Director); Rodger L. Foltz, Ph.D. (NWB Technical Director). Laboratory personnel involved in the completion o f this
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study included Suzanne Newman, B.S. (NWB Research Scientist); Laura Struhs, B.S. (NWB Scientist) and Anna Akrami, B.S. (NWB Associate Research Scientist).
NWB SOPs and guidelines were used in the conduct o f this study and were available to study personnel in electronic and/or hard copy formats.
Date Study Initiated: 23-Qct-2001 Date Analyses Completed: 16-Nov-2001
The clinical study described in this report is not included within the definition o f a GLP regulated nonclinical study. However, Northwest Bioanalytical conducts all studies within the guidelines o f the U.S. FDA Good Laboratory Practice Regulations for Nonclinical Laboratory Studies (Title 21 CFR Part 58), the OECD Principles o f Good Laboratory Practice and the Japanese MHW Good Laboratory Practice Standard Ordinance for Nonclinical Laboratory Studies on the Safety o f Drugs (Ordinance No. 21, PAB Notification No. 424). Any changes to or deviations from the original protocol (Analytical Plan) were documented through approved protocol amendments or deviation memos and are retained within the raw data.
2. M ETHODOLOGY
The assay used for this study is reported in Northwest Bioanalytical reports NWBR00108 [5.2] and NWBR00-122 [5.3]. Chinese human plasma was used for the calibration curves and quality control samples. Serum sample results obtained using plasma curves are discussed in NWBR00-122.
Samples for this study were received at NWB on the following date:
Receipt Date 23-Oct-2001
Number o fSamples Received 262
Storage Condition (except during analysis)
-20 C
)
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Reference Material
Study No. NWBSOO-145 Report No. NW BR0M55
Analyte PFOS PFOA PFHS PFOSAA PFOSA M556 M570 THPFOS
Lot Number Purity Expiration Date Source
193 100%
245 100%
SE-036
100%
617 53.8%
214 100%
NB113047-8D 99.89%
118506-26 99.75%
59909
90%
31-D ec-2010 31-D ec-2010 0 1 - J a n - 2 0 10 31-D ec-2010 31-D ec-2010 31-D ec-2010 31-D ec-2010 31-D ec-2010
3M 3M 3M 3M 3M 3M 3M 3M
*Stored dry
Storage Conditions Room temperature* Room temperature*
-20 C Room temperature Room temperature Room temperature Room temperature Room temperature
The reference material purity for PFOS, PFOSA, PFHS, and PFOA was not available prior to the conduct o f this study. Therefore, the reference material purity was assumed to be 100%.
3M contracted with Centre Analytical Laboratories, Inc., in State College, Pennsylvania to determine the absolute concentration o f PFOS, PFOA, and PFHS in NWB stock solutions used to prepare the analytical standards and controls used for this studies. All arrangements for purity determinations and the transfer o f NWB solutions to Centre Analytical Laboratories, Inc., were performed by 3M. Based on Centre Analytical Laboratories' results, the concentrations o f plasma calibrator and quality control samples were corrected using the following factors:
Analyte PFOS PFOA PFHS
Correction Factor 0.821 0.961 1.043
Eight or more calibration standards were prepared on the day of each run by adding 100 (.iL o f blank Chinese human plasma and 400 pL o f 50 mM ammonium acetate in
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water (unadjusted pH --6.9) to 13 x 100 mm polypropylene tubes. After a brief vortex mixing, 10.0 pL o f the appropriate spiking solution was added. The final calibration standard concentrations in human plasma for each analyte are listed below:
Analyte PFOS PFOA PFHS
PFOSAA PFOSA M556 M570
Calibration Standard Concentrations* 3.37,4.60, 10.8,23.1,43.6, 84.6, 208, 331 and 413 ppb 1.44, 2.88, 1 0 .1 ,2 4 .5 ,4 8 .5 ,9 7 .1 ,2 4 1 ,3 8 5 and 481 ppb 1.36, 2.92, 10.7,26.4, 52.5, 104, 261,417 and 522 ppb 1.50, 3.00, 10.5, 25.5, 50.5, 101, 251,401 and 501 ppb 1.00, 2.50, 10.0, 25.0, 50.0, 100,250, 400 and 500 ppb
2.50, 10.0, 25.0, 50.0, 100, 250, 400 and 500 ppb 1.00, 2.50, 10.0, 25.0, 50.0, 100, 250,400 and 500 ppb
* The target calibration curve range is 1.00 ppb to 500 ppb (except M556 which has a LLOQ o f 2.50 ppb). Each analyte has a different final curve range based upon the persistent levels o f the analyte in the human plasma used.
In addition, blank human plasma samples, both with and without internal standard (designated as QCOs and Blanks, respectively), were assayed in each analytical run.
Analytical QCs were prepared in Chinese human plasma on November 5, 2001 and stored in a -20 C freezer. For each run, analytical QC levels were assayed in duplicate.
A nalyte PFOS PFOA PFHS
PFOSAA PFOSA M556 M570
QC Concentrations (ppb)* Low M edium High 5.83 126 331 4.32 144 384 4.48 156 417 4.50 151 401 4.00 150 400 4.00 150 400 4.00 150 400
* The target QC concentrations are 4.00 ppb, 150 ppb and 400 ppb. Each analyte has different QC concentrations based upon the persistent levels o f the analyte in the human plasma used.
The internal standard (THPFOS) was added to all serum samples (except Blanks) for a final concentration o f approximately 200 ppb.
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The analytical method consisted o f a liquiddiquid extraction procedure followed by evaporation and reconstitution o f the extract residue with 20 mM ammonium acetate in water: 20 mM ammonium acetate in methanol (30:70 v/v). The samples were analyzed by liquid chromatography/tandem mass spectrometry using an API 3000. The instrument was operated in the multiple reaction monitoring (MRM) mode under optimized conditions for PFOS, PFOA, PFHS, PFOSAA, PFOSA, M556 and M570 detection of the negative ions formed by turboionspray ionization.
3. SAM PLE ANALYSIS
PFOS, PFOA, PFHS, PFOSAA, PFOSA, M556, M570, and THPFOS chromatographic peaks were integrated using the MacQuan software (version 1.6) with a smooth factor o f one. Quantitation was based upon quadratic regression analysis o f weighted (1/x2) 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. A cceptance C riteria
For an analytical run to be accepted, it must have met the acceptance criteria listed below which are consistent with regulatory and industry recommendations.
Calibration Curve
Each run will include in duplicate calibration standards at six or more concentrations covering the lower to upper limit o f quantitation. For all analytes except PFOSA and PFOSAA, at least three-fourths o f the calibration standard's back-calculated concentrations must be within 15% ( 20% for LLOQ) o f their individual target concentrations. For PFOSA and PFOSAA, at least three-fourths o f the calibration standard's back-calculated concentrations must be within 20% ( 25% for LLOQ) 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.
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Lower Limit o f Quantitation
Study No. NWBSOO-145 Report No. NWBROM55
The back-calculated concentrations o f at least one o f the duplicate lowest points in the calibration curve must be within 25% o f the target concentration for PFOSA and PFOSAA to qualify as the LLOQ and within 20% o f the target concentration to qualify as the LLOQ for all other analytes. If this criterion is not met, the next level is subjected to the same test and the LLOQ raised accordingly.
Quality Control Samples
Each analytical run will include low, medium and high QC samples in duplicate. The measured concentrations o f at least two-thirds o f all analytical QCs must be within 20% o f their target concentrations ( 25% for PFOSA and PFOSAA), and no two QCs at the same concentration can be outside the limit. If study samples require dilution, a dilution QC will be analyzed in triplicate for each dilution level (except for PFOSA and PFOSAA which should not be diluted with control matrix). At least two dilution QCs at each level must be within 20% o f its target concentrations in order to accept diluted study samples at that level. The dilution QC acceptance is independent o f the undiluted analytical QC acceptance.
4. RESULTS AND DISCUSSION
Serum sample results obtained using plasma curves for PFOA, PFHS, PFOSAA, PFOSA, M556, and M570 did not meet NWB SOP requirements for validation. Results may vary on average up to 26% from results obtained using human serum calibration curves at some concentration levels for PFOA, PFHS, PFOSAA, M556, and M570. For PFOSA, results may vary on average up to 43% from results obtained using human serum calibration curves. These data are reported in Assay Revalidation Addendum Report NWBROO-122 [5.3]. However, in order to obtain the lower limit of quantitation o f less than 5 ppb required by the sponsor, it was necessary to use plasma calibration curves.
Percent theoretical and percent bias are separate calculations determined from the mean. Therefore, due to rounding, percent theoretical plus percent bias may not equal 100.0%.
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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 ID 1 2 3 4 5 6 7 8
PFOS Result
PFOA Result
PFIIS R esu lt
PFOSAA R esu lt
C om m ents
Accepted Accepted Accepted Accepted
Accepted Accepted Accepted Accepted
Accepted Accepted Accepted Accepted
N /A N/A N /A N /A Run aborted, sample preparation error
Accepted Accepted Accepted Accepted
Accepted Accepted Accepted Accepted
Accepted Accepted Accepted Accepted
Accepted Accepted Accepted Accepted
N/A = not applicable
Run ID 1 2 3 4 5 6 7 8
PFOSA R esu lt Accepted Accepted Accepted
N /A Accepted Accepted Accepted Accepted
M556 Result Accepted Accepted Accepted N /A Accepted Accepted Accepted Accepted
M570 R esu lt Accepted Accepted Accepted
N /A Accepted Accepted Accepted Accepted
C om m ents Run aborted, sample preparation error
N/A = not applicable
Any known circumstances that may have affected the quality or integrity o f the data are
included in this report.
5. REFERENCES
5.1. 3M Company Protocol EPI-0016. " Identification o f Fluorochemicals in Serum from and Elderly Population in the Seattle, W ashington Metropolitan Area."
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Study No. NWBS00-145 Report No. NWBRO1-155
5.2. C. Sakashita. "Quantitative Determination o f PFOS, PFOSA, PFOSAA, POAA, PFHS, M556 and M570 in Human Serum by LC/MS/MS." Assay Revalidation Report. NWB Study NWBS00-040. NWB Report NW BR00-108. January 24, 2001.
5.3. C. Sakashita. "Quantitative Determination o f PFOS, PFOSA, PFOSAA, POAA, PFHS, M556 and M570 in Human Serum by LC/MS/MS." Assay Revalidation Addendum Report. NWB Study NWBS00-040. NWB Report N W BR00-122. November 20, 2001.
6. DATA R E T E N T IO N
The raw data and final report for this study will be stored in the NWB Archives, 1121 East 3900 South, Salt Lake City, UT 84124 per regulations and contract agreement. After submission o f the final report to the Sponsor, remaining study samples will be stored under required conditions until confirmation o f Sample Disposition/Retum Authorization is received from the Sponsor. 3M Company will be notified concerning final disposition o f records at completion of contract obligations.
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Table 1. Calibration Curve Summary for PFOS
Quadratic weighted l/x2. All concentrations are expressed as ppb.
Run Date Run Number
A
B
O8-N0V-2OOI 1 0.000000 0.004611
O9-N0V-2OOI 2 0.000000 0.004551
I3-N0V-2OOI 3 -0.000001 0.004805
I4-N 0V-2OOI 5 0.000000 0.005168
I5-N 0V-2OOI 6 -0.000001 0.004975
I5-N 0V-2OOI 7 -0.000001 0.004756
I6-N0V-2OOI 8 0.000000 0.005736
C
0.001070 -0.000952 0.001604 0.001523 -0.001202 -0.001763 -0.000223
R- LLOQ ULOQ Squared
0.9984 3.37 413 0.9958 3.37 413 0.9974 3.37 413 0.9917 3.37 413 0.9985 3.37 413 0.9956 3.37 413 0.9964 3.37 413
Mean S.D.
%CV n
0.000000 0.004943 0.000001 0.000408
8.3 77
0.000008 0.001387 17337.5
7
0.9963 0.0023
0.2 7
A, B, and C are coefficients used to define the calibration curve.
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Table 2. Calibration Curve Summary for PFOA
Quadratic weighted l/.x2. All concentrations are expressed as ppb.
Run Date
08-Nov-2001 09-NOV-2001 13-NOV-2001 14-NOV-2001 15-NOV-2001 15-NOV-2001 16-Nov-2001
Run
A
B
C R-
N um ber
Squared
1 0.000001 0.006091 0.001738 0.9965
2 0.000001 0.005194 0.000808 0.9931
3 0.000000 0.005054 0.000831 0.9960
5 -0.000001 0.005711 0.000382 0.9966
6 0.000001 0.006132 -0.000212 0.9961
7 0.000001 0.006102 0.000361 0.9977
8 0.000000 0.006771 -0.000296 0.9970
LLOQ ULOQ
1.44 1.44 1.44 1.44 1.44 1.44 1.44
481 481 481 481 481 481 481
Mean S.D.
%CV n
0.000000 0.005865 0.000516 0.000001 0.000596 0.000696
10.2 134.9 777
0.9961 0.0015
0.2 7
A, B, and C are coefficients used to define the calibration curve.
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Tabic 3. Calibration Curve Summary for PFHS
Quadratic weighted 1/x2. All concentrations are expressed as ppb.
Run Date
Run
A
B
C
N um ber
08-NOV-2001
1 -0.000002 0.007829 0.000656
09-NOV-2001 2 -0.000002 0.008314 -0.001142
13-NOV-2001 3 -0.000001 0.005315 0.000115
14-NOV-2001 5 -0.000001 0.006211 -0.000433
15-Nov-2001
6 -0.000002 0.006849 -0.001305
lS-Nov-2001
7 -0.000002 0.006791 -0.000751
16-NOV-2001
8 -0.000002 0.006953 0.000081
R- LLOQ ULOQ Squared
0.9971
1.36 522
0.9969
1.36 522
0.9972
1.36 522
0.9960 1.36 522
0.9977
1.36 522
0.9978 1.36 522
0.9986 1.36 522
Mean S.D.
-0.000002 0.006895 -0.000397 0.000000 0.000989 0.000719
0.9973 0.0008
%CV n
0.0 14.3 -181.1 0.1
77 7
7
A, B, and C are coefficients used to define the calibration curve.
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Table 4. Calibration Curve Summary for PFOSAA
Quadratic weighted l/x \ All concentrations are expressed as ppb.
Run Date Run Number
A
08-Nov-2001 1 0.000002
09-NOV-2001 2
0.000001
13-Nov-2001 3 -0.000001
14-NOV-2001 5 -0.000001
15-NOV-2001 6
0.000001
15-NOV-2001 7 0.000002
l-Nov-2001 8 0.000003
B C RSquared
0.004883 0.002297 0.9868 0.005152 0.000251 0.9916 0.010855 -0.001647 0.9963 0.010710 -0.000618 0.9811 0.008626 -0.000767 0.9955 0.007392 -0.001672 0.9905 0.011058 -0.000963 0.9960
LLOQ
1.50 1.50 1.50 1.50 1.50 1.50 1.50
ULOQ
501 501 501 501 501 501 501
Mean S.D. %CV n
0.000001 0.000002
200.0 7
0.008382 -0.000446 0.002659 0.001376
31.7 -308.5 77
0.9911 0.0056
0.6 7
A, B, and C are coefficients used to define the calibration curve.
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Table 5. Calibration Curve Summary for PFOSA
Quadratic weighted l/x \ All concentrations are expressed as ppb.
Run Date
Run
Number
A
08-NOV-2001
1 0.000001
09-NOV-2001 2 0.000001
13-NOV-2001 3 -0.000002
M-Nov-2001 5 -0.000001
15-NOV-2001 6 0.000002
15-NOV-2001 7 0.000002
I6-N 0V-2OOI 8 0.000004
B
0.005637 0.005563 0.014503 0.013789 0.010415 0.010693 0.014421
C
0.000591 0.001213 0.002158 0.002219 0.000499 0.000700 0.003092
R- LLOQ ULOQ Squared
0.9955 1.00 500
0.9908 1.00 500
0.9896 1.00 500
0.9880 1.00 500
0.9971 1.00
500
0.9940 1.00
500
0.9962 1.00 500
Mean S.D.
%CV n
0.000001 0.000002
200.0 7
0.010717 0.003872
36.1 7
0.001496 0.001003
67.0 7
0.9930 0.0036
0.4 7
A, B, and C are coefficients used to define the calibration curve.
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Table 6. Calibration Curve Summary for M556
Quadratic weighted 1/x2. All concentrations are expressed as ppb.
Run Date
Run Number
A
08-NOV-2001 1 0.000001
09-NOV-2001
2
0.000001
13-Nov-2001 3 0.000000
14-NOV-2001 5 0.000000
15-Nov-2001 6 0.000001
15-NOV-2001 7 0.000002
16-NOV-2001 8 0.000001
B
0.006345 0.006891 0.004174 0.003962 0.003844 0.003291 0.004646
C R- LLOQ ULOQ Squared
-0.000642 0.9952 2.50 500 -0.001177 0.9950 2.50 500 -0.002780 0.9942 2.50 500 0.001886 0.9844 2.50 500 -0.002125 0.9986 2.50 500 -0.000626 0.9947 2.50 500 -0.001463 0.9970 2.50 500
Mean S.D.
% cv
n
0.000001 0.000001
100.0 7
0.004736 0.001356
28.6 7
-0.000990 0.001488
-150.3 7
0.9942 0.0046
0.5 7
A, B, and C are coefficients used to define the calibration curve.
Page 21
000619
Northwest Bioanalytical
Study No. NWBS00-145 Report No. NWBROl-155
Table 7. Calibration Curve Summary for M570
Quadratic weighted 1/x2. All concentrations are expressed as ppb.
Run Date Run Number
08-NOV-2001
1
09-Nov-2001
2
13-NOV-2001
3
14-NOV-2001
5
15-NOV-2001
6
15-NOV-2001
7
I6-N 0V-2OOI
8
A
0.000001 0.000001
-0.000003 -0.000002
0.000001 0.000001 0.000001
B
0.007316 0.007844 0.015001 0.014938 0.013060 0.011552 0.016317
C
0.000681 0.001194 0.000743 0.001637 0.001232 0.000705 0.002517
R- LLOQ ULOQ Squared
0.9969 1.00 500 0.9924 1.00 500 0.9964 1.00 500 0.9922 1.00 500 0.9940 1.00 500 0.9936 1.00 500 0.9979 1.00 500
Mean S.D. %CV n
0.000000
0.000002
0.012290 0.003564
0.001244 0.000663
0.9948 0.0023
29.0 53.3 0.2 7 7 77
A, B, and C are coefficients used to define the calibration curve.
Page 22
000620
Northwest Bioanalytical
Study No. NWBSOO-145 Report No. NWBRO1-155
Table 8. Back-Calculated Concentrations of Calibration Standards for PFOS
Quadratic weighted 1/x2. All concentrations are expressed as ppb.
Run Date
Run 3.37 4.60 10.8 23.1 43.6 84.6 208 331 413
Number
08-NOV-2001 1 3.44 4.54 10.9 23.0 43.8 84.8 212 326 424
3.42 4.37 10.9 24.4 43.3 77.8 213 342 393
09-NOV-2001
2
3.58 4.49 11.1 22.9 42.2 87.3 219 336 373
3.37 4.39 10.4 21.7 41.9 85.6 238 335 403
13-NOV-2001 3 3.57 4.80 10.8 24.9 44.1 84.8 220 338 399
3.16 4.50 10.3 22.4 41.6 81.8 215 324 410
14-NOV-2001 5 3.07 5.33 10.1 23.8 43.6 82.5 213 317 412
2.97 5.41 10.2 22.7 42.8 83.6 217 334 416
lS-Nov-2001 6 3.46 4.77 10.8 24.1 40.7 85.5 208 338 426
3.24 4.51 10.7 24.1 42.0 86.3 205 325 402
15-NOV-2001 7 3.06 4.58 10.4 23.1 41.5 77.2 209 314 443
3.43 5.09 10.9 24.8 45.4 83.7 217 323 407
16-NOV-2001 8 3.63 4.32 11.6 23.8 40.6 80.8 209 328 403
3.21 4.41 11.4 24.2 44.7 78.3 206 342 425
Mean S.D.
%CV %Bias
n
3.33 4.68 10.8 23.6 42.7 82.9 214 330 410 0.211 0.357 0.440 0.947 1.47 3.27 8.28 8.98 17.0
6.3 7.6 4.1 4.0 3.4 3.9 3.9 2.7 4.1 -1.2 1.7 0.0 2.2 -2.1 -2.0 2.9 -0.3 -0.7 14 14 14 14 14 14 14 14 14
Page 23
000621
Northwest Bioanalytieal
Study No. NWBS00-145 Report No. NWBR01-155
Table 9. Back-Calculated Concentrations of Calibration Standards for PFOA
Quadratic weighted l/x2. All concentrations are expressed as ppb.
Run Date
Run
1.44
2.88
1 0 .1
24.5
48.5
97.1
241
385
481
Number
O8 -N0 V-2 OOI
1
1.56 3.08 10.1 24.5 49.1 92.3 248 373 470
1.31 2.73 9.56 26.2 49.4 89.7 255 403 476
O9 -N0V-2OOI
2
1.71 2.73 10.2 25.1 50.4 103 251 388 444
1.26 2.73 9.55 22.5 47.2 97.5 266 390 472
I3 -N0V-2OOI
3
1.60 2.90 9.74 23.8 47.7 91.7 245 377 443
1.27 2.95 9.74 26.2 50.1 96.7 252 414 488
I4 -N0V-2OOI
5
1.44 *4.47 9.53 27.5 48.7 99.6 229 380 484
1.47 *5.21 8.81 24.9 49.4 95.7 250 401 465
I5 -N0V-2OOI
6
1.36 2.58 9.29 24.7 43.9 101 244 375 481
1.56 2.97 11.1 25.5 48.3 99.0 247 391 475
I5 -N0V-2OOI
7
1.58 2.90 9.72 25.0 48.9 91.9 246 385 489
1.33 2.77 9.74 26.1 50.8 95.6 238 389 470
I6 -N0V-2OOI
8
1.54 3.12 9.94 25.5 49.2 93.0 233 390 459
1.30 2.80 9.91 25.5 48.5 93.5 233 408 496
Mean S.D. %CV
%Bias n
1.45 2.86 9.78 25.2 48.7 95.7 246 390 472
0.145 0.160 0.513 1.19 1.69 3.94 9.83 12.4 15.7
10.0 5.6 5.2 4.7 - 3.5 4.1 4.0 3.2 3.3
0.7 -0.7 -3.2 2.9
0.4 -1.4 2.1
1.3 -1.9
14 12 14 14 14 14 14 14 14
* Sample deactivated as an outlier (not included in summary statistics)
Page 24
000622
Northwest Bioanalytical
Study No. NWBSOO-145 Report No. NWBR01-155
Table 10. Back-Calculated Concentrations of Calibration Standards for PFHS
Quadratic weighted l/x2. All concentrations are expressed as ppb.
Run Date
08-N o v-2001
Run Number
1
09-N O V -2001
2
13-N O V -2001
3
14-N O V -2001
5
15-N O V -2001
6
15-N O V -2001
7
16-N O V -2001
8
1.36
1.44 1.31 1.43 1.33 1.40 1.31 1.39 1.33 1.28 1.46 1.39 1.42 1.36 1.34
2.92
2 .9 4 2 .7 2 2 .9 7 2 .7 4 2.95 2 .9 7 *4.07 *4.42 2.93 2 .7 8 2 .7 6 2.71 3.03 2.85
1 0 .7
11.5 10.3 11.2 10.2 10.1 10.6 10.6 11.1 11.2 11.4 10.1 10.7 10.6 10.9
2 6 .4
2 9 .2 2 6 .6 2 7 .3 2 5 .2 3 0 .1 2 5 .5 2 8 .5 2 6 .6 2 6 .9 2 6 .1 2 6 .3 2 8 .5 2 6 .7 2 8 .6
5 2 .5
53.3 4 9 .3 5 3 .6 4 9 .8 5 1 .5 50.9 5 1 .2 4 9 .7 5 2 .7 4 8 .9 5 2 .4 5 5 .7 4 9 .8 5 1 .9
104
99.8 99.4 111 98.2 100 103 9 3 .9 100 105 9 9 .0 106 104 103 9 7 .8
261
264 260 282 271 262 278 304 262 263 259 253 265 264 258
417
412 420 448 398 420 401 408 398 436 409 394 428 421 426
522
540 515 499 498 509 536 521 523 533 507 516 538 508 532
Mean S.D. %CV
%Bias n
1.37 0 .0 5 5 7
4.1 0 .7 14
2 .8 6 0 .1 1 5
4 .0 -2.1
12
10.8 0 .4 7 7
4 .4 0 .9 14
2 7 .3 1.46 5 .3 3 .4 14
51.5 1.94 3.8 -1 .9 14
101 4 .2 4 4 .2 -2 .9
14
268 13.0 4 .9 2 .7 14
416 15.7 3.8 -0 .2 14
520 14.4 2.8 -0.4 14
* Sample deactivated as an outlier (not included in summary statistics)
Page 25
000623
Northwest Bioanalytical
Study No. NWBS00-145 Report No. NWBRO1-155
T able 11. B ack-C alculated C oncentrations o f C alib ratio n Standards for PFOSAA
Quadratic weighted 1/x2. All concentrations are expressed as ppb.
Run Date
Run 1.50 3.00 10.5 25.5 50.5 101 251 401 501
Number
08-NOV-2001 1 1.23 2.63 10.3 25.2 52.8 111 276 428 509
1.90 2.87 10.3 26.5 51.0 86.7 251 399 448
09-No v-2001
2
1.62 2.88 10.8 26.6 53.8 105 266 405 451
1.51 2.63 9.67 23.7 46.8 98.0 300 416 473
13-NOV-2001
3
1.61 3.00 11.0 24.9 54.1 106 266 435 457
1.41 2.94 9.72 24.7 49.7 95.7 243 415 488
14-NOV-2001
5
1.40 *4.70 8.98 27.6 48.3 102 207 386 515
1.38 4.10 8.98 26.7 52.3 99.2 269 442 489
15-NOV-2001
6
1.49 3.11
10.4 28.9 42.5
103 261
411
525
1.49 2.95 10.6 26.8 48.9 98.9 235 401 477
15-Nov-2001
7
1.49 2.67 9.62 24.0 48.9 93.3 245 361 564
1.48 3.52 10.4 27.7 53.6 108 275 376 482
I6-N0V-2 OOI
8
1.53 2.76 10.6 28.1 53.5 101 255 411 506
1.62 2.61 10.2 25.8 52.3 91.9 251 398 488 '
Mean S.D.
%cv
%Bias n
1.51 2.97 10.1 26.2 50.6 100 257 406 491
0.153 0.420 0.629 1.57 3.31
6.63 21.9 22.0 31.4
10.1 14.1 6.2 6.0 6.5 6.6 8.5 5.4 6.4
0.7 -1.0 -3.8 2.7 0.2 -1.0 2.4 1.2 -2.0
14 13 14 14 14 14 14 14 14
* Sample deactivated as an outlier (not included in summary statistics)
Page 26
OOO24
Northwest Bioanalytical
Study No. NWBSOO-145 Report No. NWBR01-155
Table 12. Back-Calculated Concentrations of Calibration Standards for PFOSA
Quadratic weighted l/x \ All concentrations are expressed as ppb.
Run Date Run 1.00 2.50 10.0 25.0 50.0 100 250 400 500
Number
O8-N0V-2OOI 1 0.955 2.45 9.66 23.7 51.9 100 266 396 511
0.992 2.91 10.0 26.0 48.4 87.5 253 413 472
O9 -N0 V-2 OOI
2
0.831 2.61 9.84 25.5 51.9 100 263 376 487
1.19 2.33 9.56 23.1 48.6 98.4 298 403 479
I3 -N0V-2 OOI
3
1.09 2.89 10.6 25.0 56.8 108 258 446 462
0.875 2.37 8.69 23.2 46.8 91.4 231 423 480
I4 -N0V-2 OOI
5
1.00 *4.45 ' 8.89 27.4 48.0 98.3 208 395 496
0.919 3.12 8.63 26.8 52.2 94.6 268 436 502
I 5 -N0 V-2 OOI
6
1.09 2.66 10.2 25.7 49.2 100 256 400 525
0.919 2.27 10.2 25.9 47.4 100 236 406 477
I5-N0V-2 OOI
7
1.04 2.45 8.98 25.0 50.5 89.4 236 423 527
0.934 2.79 9.85 27.1 54.0 94.2 272 385 466
I6-N0V-2 OOI
8
1.10 2.66 9.95 27.6 50.7 94.7 249 397 510
0.915 2.24 9.88 25.7 51.1 94.1 237 418 494
Mean S.D.
%CV %Bias
n
0.989 2.60 9.64 25.6 50.5 96.5 252 408 492
0.101 0.274 0.610 1.45 2.75 5.29 22.0 19.4 20.9
10.2 10.5 6.3 5.7 5.4 5.5 8.7 4.8 4.2
-1.1 4.0 -3.6 2.4
1.0 -3.5 0.8
2.0 -1.6
14 13 14 14 14 14 14 14 14
* Sample deactivated as an outlier (not included in summary statistics)
Page 27
O00S2&
Northwest Bioanalytical
Study No. NWBS00-145 Report No. NWBROI - 155
Table 13. Back-Calculated Concentrations of Calibration Standards for M556
Quadratic weighted 1/x2. All concentrations are expressed as ppb.
Run Date Run 2.50 10.0 25.0 50.0 100 250 400 500 Number
08-NOV-2001
1
2.26 9.38 23.4 51.9 103 259 410 507
2.75 10.3 27.3 50.5 87.1 255 405 472
09-NOV-2001
2
2.65 10.1 25.8 50.2 99.3 261
389 456
2.36 9.91 23.1 46.8 97.5 291 416 494
13-NOV-2001
3
2.59
II.1 25.2
53.6
102
276
430
476
2.43 8.53 24.1 49.7 92.5 238 404 479
14-Nov-2001
5
*4.38
8.25
30.4
51.8
107
224
396
490
2.70 7.51 23.5 54.9 101 268 431 476
15-Nov-2001
6
2.48 10.1 24.7 45.5 100 257 407 515
2.52 10.0 25.1 51.0 106 252 387 480
15-NOV-2001
7
2.31 9.28 25.4 50.5 88.4 251 390 539
2.72 10.0 24.8 54.2 103 268 388 467
I6-NOV-2001
8
2.61 9.97 26.5 46.2 102 245 401 496
2.34 10.5 26.9 45.1 97.5 252 421 490
Mean S.D.
%CV %Bias
n
2.52 9.64 25.4 50.1 99.0 257 405 488
0.167
0.964
1.92
3.18 - 6.01
16.3
14.9 21.4
6.6 10.0 7.6 6.3 6.1 6.3 3.7 4.4
0.8 -3.6 1.6
0.2 - 1.0 2.8
1.3 -2.4
13 14 14 14 14 14 14 14
* Sample deactivated as an outlier (not included in summary statistics)
Page 28
00062
Northwest Bioanalytical
Study No. NWBSOO-145 Report No. NWBRO1-155
T able 14. B ack-C alculated C oncentrations of C alibration S tan d ard s for M570
Quadratic weighted l/x2. All concentrations are expressed as ppb.
Run Date
Run
LOO 2.50
10.0 25.0
50.0
100
250
400
500
Number
O8 -N 0 V-2 OOI
1
1.03 2.47 9.84 23.1 50.9 108 263 417 511
0.987 2.42 10.2 26.3 50.2 89.3 254 397 462
O9-N0V-2 OOI
2
1.11 2.52 10.1 25.9 51.4 104 264 403 443
0.925 2.27 9.96 23.2 45.4 95.9 299 411 487
I3 -N0V-2OOI
3
1.08 2.64 10.0 24.1 52.3 102 264 421 450
0.896 2.58 9.38 24.0 49.6 96.1 255 425 484
I4 -N0V-2OOI
5
0.985 *4.20 9.07 28.9 47.3 103 212 399 506
1.03 *3.85 8.56 27.0 52.2 99.8 275 424 479
I5 -N0V-2OOI
6
1.10 2.64 9.91 27.3 41.3 100 271 408 514
0.906 2.32 10.1 26.8 48.2 102 243 394 475
15-NOV-2001
7
0.930 2.33 9.10 25.1 49.4 90.7 239 373 561
1.07 2.72 9.95 26.7 52.6 105 276 382 470
I6 -N0 V-2 OOI
8
1.06 2.57 9.97 26.8 49.7 98.8 250 409 506
0.963 2.28 10.1 25.3 51.9 92.0 244 406 489
Mean S.D. %CV
%Bias n
1.01 2.48 9.73 25.8 49.5 99.0 258 405 488
0.0736 0.155 0.499 1.69 3.13 5.60 20.7 15.2 30.3
7.3 6.3 5.1 6.6 6.3 5.7 8.0 3.8 6.2
1.0 -0.8 -2.7 3.2 -1.0 -1.0 3.2
1.3 -2.4
14 12 14 14 14 14 14 14 14
* Sample deactivated as an outlier (not included in summary statistics)
Page 29
0004S27
Northwest Bioanalytical
Study No. NWBS00-145 Report No. NWBROl-155
Table 15. Analytical QC Summary for PFOS
All concentrations are expressed as ppb.
Run Date
Run Number
Preparation Date
O8-N0V-2OOI
1
09-Nov-2001
2
I 3-N 0V-2 OOI
3
H-Nov-2001
5
I5-N 0V-2OOI
6
I 5 -N 0V-2 OOI
7
16-NOV-2001
8
Low QC 5.83 ppb O5 -N0V-2OOI
5.89 5.75 6.63 6.50 6.21 6.28 5.48 5.95 6.16 6.20 5.56 6.11 6.21 6.17
M edium QC 126 ppb
05-Nov-2001 121 127 120 121 134 118 108 114 128 118 114 117 124 120
High QC 331 ppb O5 -N0V-2OOI
336 349 338 333 363 335 303 322 338 347 323 355 330 323
Mean S.D.
%CV %Theoretical
%Bias n
6.08 0.325
5.3 104.3 4.3
14
120 6.56 5.5 95.2 -4.8 14
335 15.3 4.6 101.2 1.2 14
)
Page 30
00062
Northwest Bioanalytical
Study No. NWBSOO-145 Report No. NWBROl-155
Table 16. Analytical QC Summary for PFOA
All concentrations are expressed as ppb.
Run Date
Run Low QC Medium QC High QC
Number 4.32 ppb
144 ppb
384 ppb
Preparation Date
OS-Nov-2001 05-NOV-2001 05-NOV-2001
08-NOV-2001
1
4.04
138
384
3.80 141 395
09-NOV-2001 ' 2
4.25
137
391
3.78 138 382
13-Nov-2001
3
4.03
156
436
4.46 141 430
H-Nov-2001
5
4.16
132- 351
4.63 135 391
15-NOV-2001
6
4.30
139
394
4.30 136 396
15-Nov-2001
7
4.25
140
379
4.65 139 399
l-Nov-2001
8
4.30
142
383
4.33 131 374
Mean S.D.
%CV %Theoretical
%Bias n
4.23 . 0.261
6.2 97.9 -2.1 14
139 5.89 4.2 96.5 -3.5 14
392 21.3 5.4 102.1 2.1 14
Page 31
000629
Northwest Boanalytical
Study No. NWBS00-145 Report No. NWBR01-I55
Table 17. Analytical QC Summary for PFHS
All concentrations are expressed as ppb.
Run Date
Run Number
Low QC 4.48 ppb
M edium QC 156 ppb
High QC 417 ppb
Preparation Date
05-NOV-2001 0 5 -Nov-2001 05-NOV-2001
08-NOV-2001
1
4.50
140
404
4.13 146 409
09-NOV-2001
2
4.55
140
401
4.61 142 389
13-NOV-2001
3
*6.21
176
443
4.42 144 433
14-NOV-2001
5
4.82
131
358
4.15 143 385
15-NOV-2001
6
4.58
144
397
4.27 129 408
15-Nov-2001
7
4.24
143
407
4.51 149 415
l-Nov-2001
8
4.49
141
407
4.39 144 392
Mean S.D.
%CV %Theoretical
%Bias n
4.57 0.526 11.5 102.0
2.0 14
144 10.7 7.4 92.3 -7.7 14
* > 20% deviation from theoretical
403 20.5 5.1 96.6 -3.4
14
Page 32
000630
Northwest Bioanalytical
Study No. NWBS00-145 Report No. NWBR0I-55
Tabic 18. Analytical QC Summary for PFOSAA
All concentrations are expressed as ppb.
Run Date
Run Low QC Medium QC High QC
Number 4.50 ppb
151 ppb
401 ppb
Preparation Date
05-Nov-2001 05-Nov-2001 05-Nov-2001
08-NOV-2001
1
5.01
151
392
4.35 149 419
09-NOV-2001
2
5.06
149
400
4.31 145 401
13-NOV-2001
3
*3.34
150
446
4.79 149 453
14-NOV-2001
5
3.95
139
399
4.75 148 421
15-Nov-2001
6
5.03
159
443
4.32 149 398
lS-Nov-2001
7
4.75
139
353
4.60 129 395
16-NOV-2001
8
4.74
150
390
4.89 146 387
Mean S.D.
%CV %Theoretical
%Bias n
4.56 0.478 10.5 101.3
1.3 14
147 7.07 4.8 97.4 -2.6 14
* > 25% deviation from theoretical
407 27.0 6.6 101.5 1.5 14
Page 33
000531.
Northwest Bioanalytical
Study No. NWBSOO-145 Report'No. NWBROl-155
Table 19. Analytical QC Summary for PFOSA
All concentrations are expressed as ppb.
Run Date
Run Number
Low QC Medium QC High QC
4.00 ppb
150 ppb
400 ppb
Preparation Date
O5-N0V-2OOI O5-N0V-2OOI O5-N0V-2OOI
O8-N0V-2OOI
1
3.84
149
398
4.16 149 424
O9-N0V-2OOI
2
4.17
133
393
3.67 133 404
I3-N0V-2OOI
3
3.09
159 446
4.22 153 427
I4-N0V-20OI
5
3.37
140
411
4.42 139 443
15-Nov-2001
6
4.73
151 412
3.80 142 395
I5-N0V-2OOI
7
3.82
131
346
3.80 121 399
I 6-N0V-2OOI
8
4.15
142 347
3.77 135 359
Mean S.D.
%CV %Theoretical
%Bias n
3.93 141 400 0.420 10.3 31.7 10.7 7.3 7.9 98.3 94.0 100.0 -1.8 -6.0 0.0
14 14 14
Page 34
000632
Northwest Bioanalytical
Study No. NWBS00-145 Report No. NWBR01-155
Table 20. Analytical QC Summary for M556
All concentrations are expressed as ppb.
Run Date Preparation Date 08-NOV-2001 09-NOV-2001 13-Nov-2001 14-NOV-2001 15-NOV-2001 15-NOV-2001 1-Nov-2001
Run Number
1 2 3 5 6 7 8
Low QC 4.00 ppb 05-Nov-2001
3.80 4.02 4.58 4.18 *3.07 3.41 3.43 3.38 *5.10 3.95 3.91 4.16 4.53 4.53
Medium QC 150 ppb
05-NOV-2001 140 145 145 143 151 146 148 143 157 148 148 138 145 143
High QC 400 ppb 05-NOV-2001
396 432 408 414 470 436 383 419 414 416 387 419 404 387
Mean S.D. %CV
%Theoretical %Bias n
4.00 0.564 14.1 100.0 0.0
14
146 4.70 3.2 97.3 -2.7 14
* > 20% deviation from theoretical
413 23.0 5.6 103.3 3.3 14
Page 35
000633
Northwest Bioanalytical
Study No. NWBSOO-145 Report No. NWBROl-155
Table 21. Analytical QC Summary for M570
All concentrations are expressed as ppb.
Run Date
Run Low QC Medium QC High QC
Number 4.00 ppb
150 ppb
400 ppb
Preparation Date
05-Nov-2001 O5-N0V-2OOI O5-N0V-2OOI
O8-N0V-2OOI
1
4.07
149
397
3.86 146 433
O9-N0V-2OOI
2
4.55
146
405
4.20 140 404
I3-N0V-2OOI
3
*2.95
143
451
4.32 149 455
I4-N0V-2OOI
5
3.83
142
388
4.49 146 429
I 5-N0V-2OOI
6
4.49
153
432
3.74 147 411
I5-N0V-2OOI
7
4.02
141
367
4.22 132 399
I6-N0V-2OOI
8
4.25
149
403
4.32 144 391
Mean S.D. %CV %Theoretical %Bias
n
4.09 0.415 10.1 102.3 2.3
14
145 5.13 3.5 96.7 -3.3 14
* > 20% deviation from theoretical
412 25.0 6.1 103.0 3.0 14
Page 36
000634
Northwest Bioanalytical
Study No. NWBS00-145 Report No. NWBRO1-155
Tabic 22. Scrum Study Sample Concentrations
All concentrations are expressed as ppb.
Sample Name 11331 11332 11333 11334 11335 11336 11337 11338 11339 11340 11341 11342 11343 11344 11345 11346 11347 11348 11349 11350 11351 11352 11353 11354 11355 11356 11357 11358 11359 11360 11361 11362 11363 11364 11365 11366
PFOS
33.7 28.8 44.8 12.4 18.5 25.4 29.8 33.2 32.3 37.4 35.3 83.2 30.8 61.9 54.9 34.6 58.9 57.7 14.5 24.5 14.7 13.5
22.1
71.3 48.3 59.7 18.8 25.2 49.7 19.0 16.3 95.2 133 49.1 104 39.6
PFOSA*
PFOSAA*
PFOA*
PFHS*
VI556*
M570*
<LLOQ (1.00)
1.96
6.20 <LLOQ (1.36) <LLOQ (2.50)
1.30
<LLOQ(I.00) <LLOQ (1.50)
4.73
4.50 <LLOQ (2.50) 1.37
<LLOQ ( 1.00)
3.33
6.16
3.99 <LLOQ (2.50) 2.22
<LLOQ(1.00)
1.64
2.47 <LLOQ (1.36) <LLOQ (2.50) 1.82
<LLOQ (1.00) <LLOQ ( 1.50) <LLOQ ( 1.44) <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00)
<LLOQ(1.00) <LLOQ (1.50)
2.93
2.67 <LLOQ (2.50) 1.36
<LLOQ(1.00) <LLOQ (1.50)
3.56
3.23 <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.00) <LLOQ ( 1.50)
4.03
1.73 <LLOQ (2.50) 1.72
<LLOQ(1.00) <LLOQ (1.50)
4.67
2.48 <LLOQ (2.50) 1.29
<LLOQ (1.00)
2.41
3.96 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.00)
1.50
2.94
7.19 <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.00)
19.0
4.69
3.20
4.79
3.25
<LLOQ (1.00) <LLOQ (1.50)
3.78
2.00 <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.00)
10.7
7.35
3.35
3.04
2.68
<LLOQ (1.00)
1.68
11.4
5.00 <LLOQ (2.50) 2.50
<LLOQ (1.00) <LLOQ (1.50)
4.66
1.92 <LLOQ (2.50) 1.18
<LLOQ (1.00)
5.43
7.52
7.78 <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.00)
2.44
5.01
5.80 <LLOQ (2.50) 1.18
<LLOQ (1.00) <LLOQ (1.50) 2.84 <LLOQ (1.36) <LLOQ (2.50) 1.43
<LLOQ (1.00) <LLOQ (1.50)
4.12
1.77 <LLOQ (2.50)
1.38
cLLOQ(l.OO) <LLOQ (1.50)
10.6 <LLOQ (1.36) <LLOQ (2.50)
3.55
<LLOQ (1.00)
4.73
2.37 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00)
cLLOQ(l.OO)
2.48
3.75
5.26 <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.00)
3.80
12.5
15.9 <LLOQ (2.50) 2.47
<LLOQ (1.00)
4.88
4.01 <LLOQ (1.36) <LLOQ (2.50) 1.13
<LLOQ (1.00) <LLOQ (1.50)
9.19
2.40 <LLOQ (2.50) 1.26
<LLOQ (1.00)
2.01
4.03
2.10 <LLOQ (2.50)
1.08
<LLOQ ( 1.00) <LLOQ (1.50)
1.94
10.3 <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.00)
1.93
6.32
8.53 <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.00)
4.58
4.27
1.99 <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.00) <LLOQ (1.50)
2.58
1.69 <LLOQ (2.50) cLLOQ(l.OO)
<LLOQ ( 1.00)
17.7
8.56
3.29
2.80
1.82
<LLOQ (1.00)
16.2
16.7 <LLOQ (1.36) 2.89
1.49
<LLOQ ( 1.00)
5.30
5.24
2.48 <LLOQ (2.50) 3.76
<LLOQ ( 1.00)
1.95
9.16
2.13 <LLOQ (2.50) 2.74
<LLOQ (1.00)
1.63
5.13
4.42 <LLOQ (2.50) 2.97
NOTE: The method acceptance criteria states that the measured concentration for each QC must be within 20% (25% for PFOSA and PFOSAA) of the target concentration. The statistical error associated with any individual result may exceed the QC acceptance criteria.
* Serum sample results for PFOA, PFHS, PFOSAA, PFOSA, M556 and M570 obtained using plasma calibration curves may vary from results obtained using serum calibration curves (see Results and Discussion section).
Page 37
000635
Northwest Bioanalytical
Study No. NWBS00-I45 Report No. NWBRO1-155
Table 22. Scrum Study Sample Concentrations (Continued)
All concentrations are expressed as ppb.
Sample Name 11367 11368 11369 11370 11371 11372 11373 11374 11375 11376 11377 11378 11379 11380 11381 11382 11383 11384 11385 11386 11387 11388 11389 11390 11391 11392 11393 11394 11395 11396 11397 11398 11399 11400 11401
PFOS
PFOSA*
PFOSAA*
PFOA*
PFHS*
M556*
IVI570*
13.7 10.7 31.7 30.1 22.9 27.8 18.1 40.0 18.5
21.1
71.8 35.3
11.2
60.1 29.6 35.9 26.1 19.3 30.2 47.2 53.5 28.9 38.3 31.6 18.6 37.1 25.6 43.2 24.2 73.7 41.7 45.7 10.7
22.1
44.5
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00)
2.13
cLLOQ(l.OO)
2.25
<LLOQ ( 1.00) <LLOQ ( 1.50)
<LLOQ (1.00)
2.19
<LLOQ (1.00)
13.8
<LLOQ ( 1.00) <LLOQ (1.50)
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ(I.OO)
2.22
<LLOQ (1.00)
3.55
<LLOQ ( 1.00)
1.69
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ ( 1.00) <LLOQ (1.50)
<LLOQ ( 1.00) <LLOQ (1.50)
<LLOQ ( 1.00)
1.81
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ(1.00) <LLOQ (1.50)
<LLOQ (1.00)
8.12
<LLOQ (1.00)
2.28
<LLOQ (1.00)
14.6
<LLOQ (1.00)
1.73
<LLOQ (1.00)
1.64
<LLOQ (1.00) <LLOQ ( 1.50)
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00)
2.62
<LLOQ (1.00)
5.16
<LLOQ (1.00)
5.62
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00)
2.27
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ ( 1.00)
3.24
2.20 2.31 <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.44) <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) 2.37 1.63 <LLOQ (2.50) 2.80 3.64 1.45 <LLOQ (2.50) 1.05 2.75 1.79 <LLOQ (2.50) 1.09 3.42 1.84 <LLOQ (2.50) 1.13 2.35 <LLOQ (1.36) <LLOQ (2.50) 1.18 6.85 2.98 <LLOQ (2.50) 3.25 3.32 3.04 <LLOQ (2.50) <LLOQ (1.00) 1.81 1.36 <LLOQ (2.50) 1.48 6.34 17.5 <LLOQ (2.50) 1.77
6.19 2.21 <LLOQ (2.50) 2.45 2.89 <LLOQ (1.36) <LLOQ (2.50) 1.00
4.18 2.39 <LLOQ (2.50) <LLOQ (1.00) 2.47 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) 5.08 1.58 <LLOQ (2.50) <LLOQ (1.00) 3.70 1.59 <LLOQ (2.50) 3.01 2.90 1.96 <LLOQ (2.50) 2.46 4.24 2.49 <LLOQ (2.50) <LLOQ (1.00) 5.17 2.57 <LLOQ (2.50) <LLOQ (1.00) 5.04 2.65 <LLOQ (2.50) <LLOQ (1.00) 4.65 3.95 2.67 2.06 3.82 2.40 <LLOQ (2.50) 1.49 3.44 1.90 <LLOQ (2.50) <LLOQ ( 1.00)
1.86 2.72 <LLOQ (2.50) 1.27 2.45 1.88 <LLOQ (2.50) 1.84
3.20 1.55 <LLOQ (2.50) <LLOQ (1.00) 3.98 3.63 <LLOQ (2.50) 5.69 3.47 1.87 <LLOQ (2.50) <LLOQ (1.00) 6.92 8.28 <LLOQ (2.50) <LLOQ (1.00)
4.30 2.48 <LLOQ (2.50) 1.02
5.50 1.94 <LLOQ (2.50) <LLOQ ( 1.00) 2.44 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00) 3.00 <LLOQ (1.36) <LLOQ (2.50) 1.31 5.03 5.12 <LLOQ (2.50) <LLOQ (1.00)
NOTE: The method acceptance criteria states that the measured concentration for each QC must be within 20% (25% for PFOSA and PFOSAA) of the target concentration. The statistical error associated with any individual result may exceed the QC acceptance criteria.
* Serum sample results for PFOA, PFHS, PFOSAA, PFOSA, M556 and M570 obtained using plasma calibration curves may vary from results obtained using serum calibration curves (see Results and Discussion section).
Page 38
000636
Northwest Bioanalytical
Study No. NWBS00-I45 Report No. NTVVBR01-155
Table 22. Serum Study Sample C oncentrations (Continued)
All concentrations are expressed as ppb.
Sample Name 11402 11403 11404 11405 11406 11407 11408 11409 11410 11411 11412 11413 11414 11415 11416 11417 11418 11419 11420 11421 11422
11423 11424 11425 11426 11427 11428 11429 11430 11431 11432 11433 11434 11435 11436
PFOS
51.5 21.7 29.4 22.3 40.6 40.6 27.0 13.8 104 25.1 26.5 34.1 18.0 61.3
22.1
58.9 25.6 26.3 27.2 61.3 <LLOQ (3.37) 37.3 25.4 15.1 25.8 41.6 29.3 29.7 41.1 26.3 114 48.8 30.7 24.3 14.6
PFOSA*
PFOSAA*
PFOA*
PFiis*
M556*
M570*
<LLOQ (1.00) <LLOQ (1.50)
6.35
9.74 <LLOQ (2.50) 1.39
cLLOQ(l.OO)
2.94
4.57
L.80 <LLOQ (2.50) 2.08
<LLOQ (1.00) <LLOQ (1.50)
2.99
3.42 <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.00)
1.74
2.19
1.43 <LLOQ (2.50) 2.13
<LLOQ ( 1.00) <LLOQ (1.50)
3.56
4.57 <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.00) <LLOQ (1.50)
7.00
7.00 <LLOQ (2.50) 1.01
<LLOQ (1.00)
2.06
3.57
2.46 <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.00) <LLOQ (1.50)
4.17
2.22 <LLOQ (2.50)
1.13
<LLOQ(1.00)
2.57
10.2
2.33 <LLOQ (2.50) 4.05
<LLOQ (1.00) 3.33 <LLOQ (1.44) 1.71 <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.00) <LLOQ (1.50)
3.14
8.74 <LLOQ (2.50) 2.71
<LLOQ ( 1.00) <LLOQ (1.50)
4.44
1.40 <LLOQ (2.50) <LLOQ (1.00)
<LLOQ(1.00) <LLOQ (1.50)
3.84
6.32 <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.00)
3.79
7.53
6.43 <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.00) <LLOQ (1.50) 2.28 <LLOQ (1.36) <LLOQ (2.50) 1.44
<LLOQ (1.00) <LLOQ ( 1.50)
5.67
17.1 <LLOQ (2.50) 1.83
<LLOQ (1.00)
2.43
4.69
2.78 <LLOQ (2.50) 1.14
<LLOQ (1.00)
2.23
4.30
1.57 <LLOQ (2.50)
1.62
<LLOQ (1.00) <LLOQ (1.50)| 2.27
2.02 <LLOQ (2.50)
1.95
<LLOQ (1.00) ' 10.5 8.76 6.97 <LLOQ (2.50) 3.31
<LLOQ (1.00) <LLOQ (1.50) <LLOQ (1.44) <LLOQ (1.36) <LLOQ (2.50) <LLOQ(1.00)
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00)
5.31
<LLOQ ( 1.00) <LLOQ (1.50)
<LLOQ ( 1.00) <LLOQ (1.50)
<LLOQ ( 1.00) <LLOQ (1.50)
<LLOQ (1.00)
2.94
<LLOQ ( 1.00) <LLOQ (1.50)
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ ( 1.00)
4.63
<LLOQ(1.00) <LLOQ (1.50)
<LLOQ ( 1.00)
1.87
<LLOQ (1.00)
3.14
<LLOQ ( 1.00) <LLOQ (1.50)
4.90 6.62 3.43 2.64 8.98 5.90 4.44 4.08 2.56 14.4 9.17 6.89 3.72 2.82
3.52 <LLOQ (2.50) <LLOQ ( 1.00)
1.92 <LLOQ (2.50) 4.74
<LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00)
6.22 <LLOQ (2.50) 3.48
2.66 <LLOQ (2.50) 6.30
1.68 <LLOQ (2.50) 3.51
4.74 <LLOQ (2.50) 1.63
2.15 <LLOQ (2.50) <LLOQ ( 1.00)
1.70 <LLOQ (2.50) 3.71
2.32 <LLOQ (2.50) 3.03
7.42 <LLOQ (2.50) 6.15
2.22 <LLOQ (2.50)
1.16
1.59 <LLOQ (2.50) <LLOQ(1.00)
<LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00)
NOTE: The method acceptance criteria states that the measured concentration for each QC must be within 20% (25% for PFOSA and PFOSAA) 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 39
000637
Northwest Bioanalytieal
Study No. NWBS00-145 Report No. NWBROI-155
Table 22. Serum Study Sample Concentrations (Continued)
All concentrations are expressed as ppb.
Sample Name 11437 11438 11439 11440 11441 11442 11443 11444 11445 11446 11447 11448 11449 11450 11451 11452 11453 11454 11455 11456 11457 11458 11459 11460 11461 11462 11463 11464 11465 11466 11467 11468 11469 11470 11471 11472
PFOS
20.2
32.8 49.9 15.4 20.3
12.2
34.6 17.2 11.7 29.2 20.4 13.4 36.7 36.8 50.8 39.0 34.3 29.0 27.6 16.4 138 19.0 46.1 51.6 42.9 175 9.06 26.7 17.5 33.6 32.6 64.7 28.1
22.0
61.4 28.3
PFOSA*
PFOSAA*
<LLOQ (1.00)
5.89
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00)
8.95
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00) <LLOQ (1.50)
cLLOQ(l.OO)
2.26
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00) <lLOQ (1.50)
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00)
3.76
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ(1.00) <LLOQ (1.50)
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00)
2.86
<LLOQ ( 1.00)
2.18
<LLOQ (1.00)
1.62
<LLOQ(1.00)
2.40
<LLOQ (1.00)
21.1
<LLOQ ( 1.00)
1.57
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ ( 1.00)
2.81
<LLOQ (1.00)
1.60
<LLOQ ( 1.00)
2.46
<LLOQ (1.00)
1.79
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00)
2.00
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ ( 1.00)
1.78
<LLOQ (1.00)
2.77
<LLOQ ( 1.00)
1.77
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ ( 1.00)
6.75
<LLOQ(1.00) <LLOQ (1.50)
PFOA*
2.63 6.47 5.96 3.33 4.30 1.84 4.54 2.33 1.87 3.45 2.99 2.78 4.05 3.18 4.87 2.85 4.44 4.19 2.48 3.42 11.3 2.76 4.81 6.17
2.21
9.25 4.35 3.02 2.99 4.40 5.41 6.84 3.96 1.81 9.95 4.22
PFHS*
M556*
M570*
<LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00)
3.22 <LLOQ (2.50) 1.55
<LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00)
2.05 <LLOQ (2.50) 1.28
2.15 <LLOQ (2.50) <LLOQ ( 1.00)
2.65 <LLOQ (2.50) <LLOQ (1.00)
4.25 <LLOQ (2.50) 1.70
<LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00)
3.71 <LLOQ (2.50) <LLOQ ( 1.00)
2.30 <LLOQ (2.50) <LLOQ ( 1.00)
<LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.36) <LLOQ (2.50)
1.18,
2.69 <LLOQ (2.50) 2.86
1.72 <LLOQ (2.50) 2.58
1.80 <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.36) <LLOQ (2.50) <LLOQ ( 1.00)
1.47 <LLOQ (2.50) <LLOQ ( 1.00)
<LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.36) <LLOQ (2.50)
1.16
<LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.36)
2.82
1.22
<LLOQ (1.36) <LLOQ (2.50)
2.86
5.25 <LLOQ (2.50) <LLOQ (1.00)
1.58 <LLOQ (2.50) 1.52
5.44 <LLOQ (2.50) 4.94
17.0 <LLOQ (2.50) 1.01
4.90 <LLOQ (2.50) 1.02
2.74 <LLOQ (2.50) 1.71
<LLOQ (1.36) <LLOQ (2.50)
2.00
4.23 <LLOQ (2.50) 1.75
4.00 <LLOQ (2.50) 2.81
1.59 <LLOQ (2.50) 4.23
2.16 <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00)
1.64 <LLOQ (2.50) 2.87
3.56 <LLOQ (2.50) <LLOQ (1.00)
NOTE: The method acceptance criteria states that the measured concentration for each QC must be within 20% (25% for PFOSA and PFOSAA) 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 40
000638
Northwest Bioanalytical
Study No. NWBSOO-145 Report No. NWBRO1-155
Table 22. Serum Study Sample Concentrations (Continued)
All concentrations are expressed as ppb.
Sample ame 11473 11474 11475 11476 11477 11478 11479 11480 11481 11482 11483 11484 11485 11486 11487 11488 11489 11490 11491 11492 11493 11494 11495 11496 11497 11498 11499 11500 11501 11502 11503 11504 11505 11506 11507 11508
PFOS
PFOSA*
PFOSAA*
32.1 27.9 27.4 18.9 38.1 47.5 28.4 48.4 26.0 19.7 47.2 72.5 33.6 31.8 37.3 25.2 21.4 29.6 91.5 50.2 24.4 47.0 48.6 47.3 76.4 34.2 57.0 20.5 17.2 42.2 56.1 30.5 104 44.0 32.1 24.3
cLLOQ(l.OO) <LLOQ (1.50)
<LLOQ(1.00) <LLOQ (1.50)
cLLOQ(l.OO) <LLOQ (1.50)
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ ( 1.00)
1.96
cLLOQ(l.OO)
1.61
<LLOQ (1.00) <LLOQ (1.50)
cLLOQ(l.OO)
6.49
cLLOQ(l.OO) <LLOQ (1.50)
<LLOQ(1.00) <LLOQ (1.50)
cLLOQ(l.OO)
2.50
<LLOQ(1.00)
7.66
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00)
3.40
<LLOQ (1.00)
1.68
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00)
2.67
<LLOQ (1.00)
1.85
cLLOQ(l.OO) <LLOQ (1.50)
<LLOQ (1.00)
1.82
<LLOQ ( 1.00)
4.38
<LLOQ (1.00)
1.53
<LLOQ ( 1.00)
1.82
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00)
2.57
<LLOQ. (1.00)
5.54
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00)
1.58
<LLOQ ( 1.00) <LLOQ (1.50)
<LLOQ (1.00)
2.29
<LLOQ (1.00)
2.10
<LLOQ (1.00)
2.31
<LLOQ (1.00) <LLOQ (1.50)
PFOA*
PFHS*
M556*
M570*
3.60
6.01
3.59 3.21 6.16 6.97 4.44 7.22 4.74 3.15 6.41 7.20 4.21 3.69 6.65 5.40 3.01 7.23 10.5 4.16 4.75 5.95 7.78 4.62 8.95 3.72 5.38 3.96
2.21
7.45 6.90 3.52 7.81 8.07 3.89 2.57
2.67 <LLOQ (2.50)
1.67
3.66 <LLOQ (2.50) 2.38
5.79 <LLOQ (2.50) 1.09
<LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00)
2.38 <LLOQ (2.50) 1.84
3.19 <LLOQ (2.50) 1.52
6.50 <LLOQ (2.50) 1.99
1.69 <LLOQ (2.50)
1.54
3.64 <LLOQ (2.50)
1.70
<LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00)
2.06 <LLOQ (2.50)
1.43
<LLOQ (1.36)
4.09
1.82
3.72 <LLOQ (2.50) 1.08
1.95 <LLOQ (2.50) <LLOQ (1.00)
3.21 <LLOQ (2.50) 3.39
3.78 <LLOQ (2.50) 1.43
<LLOQ (1.36) <LLOQ (2.50)
1.41
<LLOQ (1.36) <LLOQ (2.50)
3.44
5.42 <LLOQ (2.50)
1.50
3.12 <LLOQ (2.50) 1.67
2.47 <LLOQ (2.50) 1.53
2.62 <LLOQ (2.50) 2.23
3.15 <LLOQ (2.50) 1.48
5.50 <LLOQ (2.50) <LLOQ ( 1.00)
2.27 <LLOQ (2.50) 1.44
2.77 <LLOQ (2.50)
1.45
10.6 <LLOQ (2.50)
1.78
<LLOQ (1.36) <LLOQ (2.50)
1.60
<LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00)
2.55 <LLOQ (2.50) 1.97
40.3 <LLOQ (2.50) 6.56
3.94 <LLOQ (2.50) 1.10
3.85 <LLOQ (2.50) 2.08
6.92 <LLOQ (2.50) 1.90
2.21 <LLOQ (2.50) 1.01
<LLOQ (1.36) <LLOQ (2.50)
1.49
NOTE: The method acceptance criteria states that the measured concentration for each QC must be within 20% (25% for PFOSA and PFOSAA) of the target concentration. The statistical error associated with any individual result may exceed the QC acceptance criteria.
* Serum sample results for PFOA, PFHS, PFOSAA, PFOSA, M556 and M570 obtained using plasma calibration curves may vary from results obtained using serum calibration curves (see Results and Discussion section).
Page 41
60063*?
Northwest Bioanalyticai
Study No. NWBSOO-145 Report No. NWBROI-155
Table 22. Serum Study Sam ple Concentrations (Continued)
All concentrations are expressed as ppb.
Sample Name 11509 11510 11511 11512 11513 11514 11515 11516 11517 11518 11519 11520 11521 11522 11523 11524 11525 11526 11527 11528 11529 11530 11531 11532 11533 11534 11535 11536 11537 11538 11539 11540 11541 11542 GH11281 GH11282
PFOS
22.9 46.7 161 54.4 14.5 20.4 26.5 37.6 49.2 8.50 19.5 25.3 18.6 35.2 28.7
20.1
23.9 49.8 15.4 31.1 15.6 19.3
21.2
40.4 25.5 20.5 23.2 38.6 15.4 33.4 32.9 15.7 24.4 40.9 36.2 18.2
PFOSA*
PFOSAA*
<LLOQ (1.00)
1.59
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00)
5.99
<LLOQ ( l .00) <LLOQ (1.50)
<LLOQ (1.00)
3.03
<LLOQ ( 1.00)
1.58
<LLOQ (1.00) <LLOQ ( 1.50)
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ ( 1.00) <LLOQ (1.50)
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ(1.00) <LLOQ ( 1.50)
<LLOQ (1.00)
1.92
<LLOQ ( 1.00)
1.83
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00)
1.80
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ(1.00) <LLOQ (1.50)
<LLOQ(1.00)
2.30
<LLOQ ( 1.00) <LLOQ (1.50)
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00)
1.62
<LLOQ ( 1.00) <LLOQ (1.50)
cLLOQ(l.OO) <LLOQ (1.50)
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00)
1.86
cLLOQ(l.OO) <LLOQ (1.50)
<LLOQ ( 1.00) <LLOQ (1.50)
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ ( 1.00) <LLOQ (1.50)
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ ( 1.00) <LLOQ (1.50)
cLLOQ(l.OO) <LLOQ (1.50)
<LLOQ (1.00)
1.68
PFOA*
5.53 6.62 14.2 5.07 4.58
2.12
2.97 5.83 9.56 1.57
2.10
3.63 2.48 5.36 4.73 3.48 3.61 5.51 2.82 4.43 2.15 3.18 2.99 4.76
1.88 2.20
2.62 6.95 2.43 2.38 4.40 2.25 3.69 3.77 5.91 2.58
PFHS*
M556*
M570*
2.23 <LLOQ (2.50) <LLOQ (1.00)
5.35 <LLOQ (2.50) 1.13
4.61 <LLOQ (2.50) <LLOQ (1.00)
3.30 <LLOQ (2.50) 2.87
8.03 <LLOQ (2.50) 1.20
<LLOQ (1.36) <LLOQ (2.50)
2.86
4.86 <LLOQ (2.50) 1.54
4.35 <LLOQ (2.50)
1.05
2.03 <LLOQ (2.50) 2.48
3.72 <LLOQ (2.50) <LLOQ (1.00)
2.35 <LLOQ (2.50) 1.53
<LLOQ (1.36) <LLOQ (2.50)
2.70
1.71 <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.36) <LLOQ (2.50) <LLOQ(1.00)
2.39 <LLOQ (2.50) 1.21
1.37 <LLOQ (2.50) <LLOQ (1.00)
1.38 <LLOQ (2.50) <LLOQ (1.00)
6.41 <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.36) <LLOQ (2.50) <LLOQ ( 1.00)
3.99 <LLOQ (2.50) 2.05
<LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00)
2.71 <LLOQ (2.50) <LLOQ(1.00)
<LLOQ (1.36) <LLOQ (2.50)
2.46
2.16 <LLOQ (2.50) 2.57
1.46 <LLOQ (2.50) <LLOQ (1.00)
1.38 <LLOQ (2.50) <LLOQ (1.00)
1.49 <LLOQ (2.50) <LLOQ (1.00)
9.60 <LLOQ (2.50) 1.26
<LLOQ (1.36) <LLOQ (2.50)
1.45
<LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00)
2.28 <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00)
21.8 <LLOQ (2.50) 3.10
2.68 <LLOQ (2.50) 2.00
3.10 <LLOQ (2.50) 2.13
<LLOQ (1.36) <LLOQ (2.50)
2.49
NOTE: The method acceptance criteria states that the measured concentration for each QC must be within 20% (25% for PFOSA and PFOSAA) of the target concentration. The statistical error associated with any individual result may exceed the QC acceptance criteria.
* 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
000640
Northwest Bioanalytical
Study No. NWBS00-145 Report No. NWBRO1-155
Table 22. Serum Study Sample Concentrations (Continued)
All concentrations are expressed as ppb.
Sample Name GH11283 GHIL284 GH11285 G H I1286 GH11287 GHI 1288 GHI 1289 GHI 1290 GHI 1291 GHI 1292 GHI 1293 GHI 1294 GHI 1295 GHI 1296 GH11297 GHI 1298 GHI 1299 GHI 1300 GHI 1301 GHI 1302 GHI 1303 GHI 1304 GHI 1305 GHI 1306 GHI 1307 GHI 1308 GHI 1309 GHI 1310 GHI 1311 GHI 1312 GHI 1313 GHI 1314 GHI 1315 GHI 1316 GH11317 GHI 1318
PFOS
PFOSA*
PFOSAA*
PFOA*
PFHS*
M556*
M570*
13.8 18.7
21.1
29.4 52.0 40.5 37.7 22.7 41.3 62.6 9.58 20.3 16.3 30.0
21.8
54.3 15.4 27.2 27.1 15.7 30.5 16.2 11.7
21.2
36.1 35.7
21.2
44.8 103 13.4 27.4 26.0 31.8 27.1 39.7 37.2
<LLOQ (1.00) <LLOQ (1.50)
2.29
3.37 <LLOQ (2.50) <LLOQ (1.00)
<LLOQ(1.00) <LLOQ ( 1.50)
3.60
5.20 <LLOQ (2.50) 1.13
<LLOQ (1.00) <LLOQ (1.50)
3.82
1.96 <LLOQ (2.50) 2.52
<LLOQ (1.00)
3.09
3.69
1.83 <LLOQ (2.50) <LLOQ ( 1.00)
<LLOQ ( l .00)
1.74
4.55 <LLOQ (1.36) <LLOQ (2.50) 4.24
<LLOQ ( 1.00) <LLOQ ( 1.50)
7.41
1.78 <LLOQ (2.50)
1.78
<LLOQ (1.00)
1.81
6.69
2.94 <LLOQ (2.50) 2.62
<LLOQ (1.00)
3.25
4.13 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.00)
1.54
4.53
8.76 <LLOQ (2.50) 1.04
<LLOQ (1.00)
2.09
6.56
5.25 <LLOQ (2.50) 3.48
<LLOQ (1.00)
5.06
2.04 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00)
<LLOQ(1.00) <LLOQ (1.50)
5.59
1.70 <LLOQ (2.50)
1.61
<LLOQ (1.00) <LLOQ ( 1.50) <LLOQ ( 1.44) 7.23 <LLOQ (2.50) 2.82
<LLOQ (1.00)
4.29
5,24
3.08 <LLOQ (2.50) 1.91
<LLOQ (1.00)
1.53
3.96 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.00)
1.68
6.52
2.84 <LLOQ (2.50) 4.11
<LLOQ ( 1.00) <LLOQ (1.50) 2.55 <LLOQ (1.36) <LLOQ (2.50) <LLOQ ( 1.00)
<LLOQ (1.00)
1.77
4.28
1.85 <LLOQ (2.50) 1.02
<LLOQ (1.00)
2.61
6.88
3.18 <LLOQ (2.50) 2.09
<LLOQ (1.00) <LLOQ (1.50) 3.17 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00)
<LLOQ ( 1.00)
1.61
3.73 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00)
<LLOQ ( 1.00)
3.75
1.79 2.03 <LLOQ (2.50) 1.49
<LLOQ (1.00)
2.24
3.44 <LLOQ (1.36) <LLOQ (2.50) 1.43
<LLOQ ( 1.00) <LLOQ (1.50) 3.22 <LLOQ (1.36) <LLOQ (2.50) 2.37
<LLOQ (1.00) <LLOQ (1.50)
3.92 . <LLOQ (1.36) <LLOQ (2.50)
1.40
<LLOQ (1.00)
2.05
3.34
1.90 <LLOQ (2.50) 2.52
<LLOQ (1.00) <LLOQ (1.50)
3.04
10.6 <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.00)
1.64
3.50
2.01 <LLOQ (2.50) 1.77
<LLOQ ( 1.00)
7.65
15.0 6.84 <LLOQ (2.50) 2.79
<LLOQ (1.00) <LLOQ (1.50) 3.47 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.00) <LLOQ (1.50)
4.08
2.74 <LLOQ (2.50) 2.12
<LLOQ ( 1.00) <LLOQ (1.50)
4.59
2.75 <LLOQ (2.50) 5.07
<LLOQ (1.00) <LLOQ (1.50)
3.81
2.11 <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.00) <LLOQ (1.50)
3.86
. 2.78
<LLOQ (2.50)
1.59
<LLOQ ( 1.00)
2.06
5.33
3.90 <LLOQ (2.50) 1.75
<LLOQ ( 1.00)
1.66
3.56 <LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00)
NOTE: The method acceptance criteria states that the measured concentration for each QC must be within 20% (25% for PFOSA and PFOSAA) 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).
00
Northwest Bioanalytical
Study No. NWBS00-145 Report No. NWBROl-155
Table 22. Scrum Study Sample Concentrations (Continued)
All concentrations are expressed as ppb.
Sample Name G H I1319 GH11320 GHI 1321 GHI 1322 GHI 1323 GHI 1324 GHI 1325 GHI 1326 GHI 1327 GHI 1328 GHI 1329 GHI 1330
PFOS
34.1
21.6
43.5 104 10.9 27.6 17.7 45.6 69.6 36.4
12.8
27.3
PFOSA*
PFOSAA*
<LLOQ(l.OO) <LLOQ (1.50)
<LLOQ (1.00) <LLOQ ( 1.50)
<LLOQ ( 1.00)
6.67
<LLOQ (1.00)
9.85
<LLOQ (1.00)
2.94
<LLOQ (1.00)
2.69
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00)
11.1
<LLOQ (1.00)
2.37
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00) <LLOQ (1.50)
<LLOQ (1.00) <LLOQ (1.50)
PFOA*
4.24 3.91 6.96 15.4
2.12
4.55 2.96 5.89 8.38 3.35 3.22 3.03
PFHS*
M556*
M570*
2.55 <LLOQ (2.50) 1.29
2.97 <LLOQ (2.50) <LLOQ (1.00)
2.19 <LLOQ (2.50) 1.04
3.73 2.75 1.06
<LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00)
3.58 <LLOQ (2.50)
1.30
<LLOQ (1.36) <LLOQ (2.50)
1.16
6.02 <LLOQ (2.50) 2.06
2.39 <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.36) <LLOQ (2.50) <LLOQ (1.00)
<LLOQ (1.36) <LLOQ (2.50)
1.53
1.92 <LLOQ (2.50) <LLOQ (1.00)
NOTE: The method acceptance criteria states that the measured concentration for each QC must be within 20% (25% for PFOSA and PFOSAA) of the target concentration. The statistical error associated with any individual result may exceed the QC acceptance criteria.
* Serum sample results for PFOA, PFHS, PFOSAA, PFOSA, M556 and M570 obtained using plasma calibration curves may vary from results obtained using serum calibration curves (see Results and Discussion section).
Page 44
000642
Northwest Bioanaiytical
Study No. NWBSOO-145 Report No. NWBROI-155
Figure 1. Representative Calibration Curve for PFOS
Analytical Run 6 analyzed on 15-Nov-2001 Calibration Standards for PFOS (ppb) Regression Method = QUADRATIC - Weighting Factor = l/X**2 Quadratic Limit - 4300
Figure 2. Representative Calibration Curve for PFOA
Analytical Run 6 analyzed on 15-Nov-2001 Calibration Standards for PFOA (ppb) Regression Method = QUADRATIC - Weighting Factor * 1/X**2 Quadratic Limit = -4270
Page 45
000643
Northwest Bioanalytical
Study No. NWBSOO-145 Report No. NWBR01-155
Figure 3. Representative Calibration Curve for PFHS
Analytical Run 6 analyzed on 15-Nov-2001 Calibration Standards for PFHS (ppb) Regression Method = QUADRATIC - Weighting Factor = 1/X**2 Quadratic Limit " 2200
p p Instalment Response
Figure 4. Representative Calibration Curve for PFOSAA
Analytical Run 6 analyzed on 15-Nov-2001 Calibration Standards for PFOSAA (ppb) Regression Method QUADRATIC - Weighting Factor" l/X**2 Quadratic Limit = -3640
ment Response
)
Page 46
000644
Northwest Bioanalytical
Study No. NWBSOO-145 Report No. NWBRO1-155
Figure 5. Representative Calibration Curve for PFOSA
Analytical Run 6 analyzed on lS-Nov-2001 Calibration Standards for PFOSA (ppb) Regression Method = QUADRATIC - Weighting Factor " 1/X**2 Quadratic Limit -2180
Figure 6. Representative Calibration Curve for M556
Analytical Run 6 analyzed on lJ-Nov-2001 Calibration Standards forM556 (ppb) Regression Method QUADRATIC - Weighting Factor = 1/X**2 Quadratic Limit * -2310
0 50 100 150 200 250 300 350 400 450 500 Nominal Cone.
Page 47
000645
Northwest Bioanalytical
Study No. NWBS00-145 Report No. NWBR01-I55
Figure 7. Representative Calibration Curve for M570
Analytical Run 6 analyzed on 15-Nov-200I Calibration Standards for M570 (ppb) Regression Method " QUADRATIC - Weighting Factor = l/X**2 Quadratic Limit = -12500
Instrument Response
Page 48
000640
Northwest Bioanalytical
Figure 8. Human Plasma Blank for PFOS
Study No. NWBSOO-145 Report No. NWBROI-155
THPfOS use is Internal Standard
I
Current Method
Noise Ttwei
20.0
Quant Thre 4.C Min. Wid ___ 5
4.0
5
Mult. Wid __10
to
Expected RT 4:3t
Base. Widt too RT Win. (sea _ 25
too 25
Smooti
S001450I037 1 037 500M S CONTROt_BtANtt4o Comment
Page 49
OOQS47
Northwest Bioanalytical
Study No. NWBS00-145 Report No. NWBROl-155
Figure 9. Human Plasma Blank for PFOA
IPFCM
I
Internai Standard: THfTOS
Ute Area
Absolute Retention Time
Expected RT 4:37
Current Method
N ote Thre>
IQ.Q
Quatti ihre
J.O
Min. Wid
8
Mult. Wid
Baie. Widt
too
RT Win. Itec
26
5mood
I
ITHPfOS
I
use as Internai Standard
apeoed RT 4:3 t
Current Method
Notte Tbret 20.0 20.0
Quant Thre 4.C 4.0
Min. Wid s 3
Mull. Wid
10 10
Base. Widt 100 100
RT Win. (sec: 25
25
Smootl 1 1
S0014S01037 1 037 S00145 CONTROt_BlANJ04o Comment
Page 50
00064S
Northwest Bioanalytieal
Study No. NWBS00-145 Report No. NWBRO1-155
Figure 10. Human Plasma Blank for PFHS
(pfh5
I
Internal Standard: THPFOS
UmArea
Absolute Retention firne
ExpectedRT 3:56
Current Method
Noise Threi 5.0 s.o QuamThre 1.5 1.5
Min. Wid 5 5 Mult. Wid 18 18 Base. Widt 100 100
RTWin. (sec SO SO Smodi 1 1
$0014501037 1 037 S0 0M 5 CONTROL BlAN IW o Commetti
VtMPfOS
1
useasInternal Standard
Current Method NoiseThr 20.0 20.0 Quant Threi 4.0 4.0
Min. Wid 5 5
Mult. Wtd 10 10
ExpeaedRT 4:3t
Base. Widi 100 100 RTWin. (sec 25 25
Smodi 1 1
500U51037 1037500145CONTROl.BLANIiNaComment
Page 5 1
000649
Northwest Bioanalytieal
Study No. NWBS00-145 Report No. NWBROl-155
Figure 11. Human Plasma Blank for PFOSAA
ITHPfOS use at Internal Standard
Expected RT 4J1
Current Method
Noise Ihre! _
20.0
Quant Thr _ Min. WW _ Mult. Wld _
4.0
3 10
Base. Widt,
100
RT Win. (se a ,
25
Smootl
S0014S01037 1 037 S0014S CONTROL BlANKNo Comment
Page 52
000650
Northwest Bioanalytical
Study No. NWBS00-145 Report No. NWBR01-155
Figure 12. Human Plasma Blank For PFOSA
[PfO S A internal Standard: THPfQS Ute Area Absolute Retention Time Expected RT 5:51
Currj Noiw Threi Quem Thre
Min. Wid Mult. Witl Bete. Widr RT Win. (
7.0
1.5 5
17
100
S0 0I4S0I037 I QJ7S0014SCONTROL.BLANI04O Comment Fri. Nov 9. 2001 3:11AM
0:59 in 1 period 1: 8:59 MRM. 642 team
497.9->77.9
Area 0 Heitht 0 Start Tinte End Time Intension Width Retention Time Intera! ion Tvoe
rtiTFRg
0:00.0 0:00.0 0:00.0 0:00.0
m
ntwjrty: I M cp*
61 'Mil : li l "241- Ml ' ; l 421 ; 0-51 1 42 2 32 223 4 17 SOI I M &4
M735;'4;i323I-.STioTeMtRii-
Page 53
000651
Northwest Bioanalytical
Study No. NWBS00-I45 Report No. NWBRO1-155
Figure 13. Human Plasma Blank for M556
M356
1
Internai Standard: THPfOS
U m Area
Abtolute Retention Time
Expected RT 5:36
Curten! Method N imm Threi S.O 5.0 Quant Thret a. 5 0.S
Min. Wid 5 S Mutt. Wld 21 21 Baie. WkW 100 100 RT Win. (sea 20 20
Smoot t 1
It m p t o s U H lntt.ulSund>d
I
Current Method
Notte Thre: 20.0 20.0
Qmm Tin*
4.0 4.0
Min. Wid S 5
Muli. Wid 10 10
Expected RT 4:31
B" - WMl 100 100 RT Win. (tee 25 25
Smootl 1 1
$0014501037 1 037 S00145 C O N T R O t. lA N W o Comment
Page 54
000652
Northwest Bioanalytical
Study No. NWBSOO-145 Report No. NWBROI-155
Figure 14. Human Plasma Blank for M570
Im jto
I
Ute Area Absolute Retention Time
Current Method Nmse Thres 5 0 5.0 Quatti tuet
Min. Wid 5 5 Mult. Wid 20 20 Base. Widt 100 100 RT Win. (sed 20 20
Smoot' 1 1
$0014501037 1 037 S0014S CONTROL BLANKNo Comment
iTHPfOS
1
umas Internal Standard
Expeoed RT 4:31
Current Method
Moite Thres _
2ao
Quant Tbne 4.0 4.0
Min. Wid ___5
5
Mult. Wid', __KJ
>00Base. Widt _ 25RTWin. {sea,
_1Smootl
10
100
25
$0014501037 1 037 $00145 CONTROl_8LAN4o Comment
Page 55
000653
Northwest Bioanalytical
Study No. NWBS00-145 Repon No. NWBROI-155
Figure 15. Human Plasma Blank with Internal Standard (QCO) for PFOS
Page 56
000654
Northwest Bioanalytica!
Study No. NWBSDO-145 Report No. NWBROl-155
Figure 16. Human Plasma Blank with Internal Standard (QCO) for PFOA
IW OA
I
lniern.il Standard: rHPfQS
Use Area
Absolute Retention Time
Expected RT 4:37
Curve Method
Noise ihre: <
10.0
Quant Thte _ 3.0 3.0
M*n. Wid 8 8
Mult. Wid 16 16
Base. Widi 100 100
RT Win. (tecs , 26
26
Smootl
11
50014501011 I O il S014S Q C O INo Comment Thu. Nov 8. 2001 10.17 PM
8:59 in 1 period 1: 8:59 MRM . 642 scant
413.0-> 169.0
Area 5879
Meiht 607
Start Time
4:23
End Time
4:51
Integration Width 0:27.7
Retention Time
4:37
Inteeration Type A BB
Itl+HH i
3
iotastty; 656eps
r$.-;V i2 risi r 241 ,3 i- 3 r:--: c i -,; m 's - i r ' v'sc^ b!Q l 2:32':`' .3:T3T4:-1S '^ 0 : : & ; 3 4 !^ 6 ; 4 + : ^ ^
ITHPFOS
"
use as internai Standard
I
Expected RT 4:31
Current Method
Noise Thre*
20.0
Quant Ihre Min. Wid
4.0
5
Muli. WW
to
Base. WWt
100
RT Win, (secs
2S
Smood
SO0US01011 t O il SO O I4SO CO INO Comment
Page 57
090155
Northwest Bioanalytical
Study No. NWBS00-145 Report No. NWBRO1-155
Figure 17. Human Plasma Blank with Internal Standard (QCO) for PFHS
im is '
I
Internal Standard: THPfOS
Ute Area
Absolute Retention Time
Expected RT 3:56
Current Method
Noise Ihres
s.o
1.5Quant Thre 1.5
Min. Wid Muk. Wid Base. Widt
5___ m
too
5 18
100
HT win. (tea 50 50
Smooti
I
SOO14SOI0M t o n S00I45 Q C O INO Comment
ITHPfOS
I
use as Internal Standard
Expected RT 4:31
Current Method
Noise Thre
20.0
Quam Thre 4.0 4.0
10 10Mm. Wid ___ 5
Mult. Wid '
5
Base. Widt 100 100
RT Win. (sea < 25
25
Smooti
\
5001450101) I 011 $00145 Q C O INo Comment
Page 58
009651
Northwest Bioanalytical
Study No. NWBS00-I45 Report No. NWBR01-155
Figure 18. Human Plasma Blank with Internal Standard (QCO) for PFOSAA
iPfOSAA
1
Intimai Standard: THPFOS
Uw Area
Absolute Reteniion Time Expected RT 6:03
Current Method
Note Thrt^ 5.0 S.O
Quant Thre 1.3 t.J
Min. Wid 6 6
Mult. W h) J7 17
Bate. Wid? 1001 100
RT Win. (tecs 30
20
Smootl 1 1
S0QV4S01QU 1 O U SOQ1*5 O C O IN o Comment
Thu. N o * fl. 200t U h l7 P M
0:59 in 1 period
TOOr .:
1: 8:59 MRM, 642 scans
54.0*>4I9.0
Area 4592 Heithi 590 Stau Time End Time Imexratlon Width Retention Time Integration Type
5:33 6:18 0:25.2 6:07 A B8
inteftiity:6l9eH 437
204 125160
W )323
270
kmtfffiL
SSI
' 6 -* i r ^ *18 - 241f 3(Jl ':.0'51Kt:-43- '.232' rS23
'421^401
SOI > W n w
. ^ - x . 6:44 ,735 25- T * m
ITHPfOS uw as Internal Standard
Expected RT 4:31
Current Method
Noise Thre 20.C 20.0
Quant Ihre 4.C 4.0
Min. WM 5 5
Mult. Wid 10 10
Base. Widt 100 100
RT Win. (sea 25
25
Smootl t 1
S0014501011 1 O ll S00145O C O IN o Comment
Page 59
000657
Northwest Bioanalytical
Study No. NWBSOO-145 Report No. NWBRO1-155
Figure 19. Human Plasma Blank with Internal Standard (QCO) for PFOSA
|F*0$A
1
Internal Standard: TMPFOS
Ute Area
Absolute Retention Time
Expected RT 5:51
Current Method
Notte Hues 7.0 7.0
Quant Thre 1.5 I.S
Min. Wid S 5
Muli. Wid 17 i ;
Bate. WUi I X
IX
RT Win. (tee 20 20
Smootl
1
$001430101J 1 OM $00143 Q C O INo Comment Thu. Nov 8. 2 X 1 10:17PM
8:39 in 1 period
1: 8:59 MRM . 642 scant
497.9->77.9
Area 0
Heitht 0
Stari Urne
000.0
End Time
o-.x.o
Integration Width 0:00.0
Retention Time
000.0
Integration Tvpe
ETCFHB m
Page 60
000658
Northwest Bioanalytical
Study No. NWBSOO-145 Report No. NWBROl-155
Figure 20. Human Plasma Blank with Internal Standard (QCO) for M556
laiss
I
Internal Standaid: fHPFOS
Use Area
Absolute Retention Time
Expected RT 5:36
Current Method
Noise Thres $.0 S.O
Quant Thre
0.S
Min. WuJ 5 5
Muil. Wid 21 21
8ase. Widt 100 100
RT Win. (see 20
20
Smooti 1 1
SOOt45010t t I 011 $00145 Q C O INo Comment
(THPfOS
\
uw as Internal Standard
Expected RT *:31
Current Method
Notte Thr
20.0
Quant Thre Min. Wld
4.0
5
Mult. W.d a
10
Base. Wld
too
RT Win. (tecs,
25
Smootl
$0014501011 t O il $00145 Q C O IN o Comment
Page 61
000659
Northwest Bioanalytical
Study No. NYVBS00-I45 Report No. NWBRO1-155
Figure 21. Human Plasma Blank with Internal Standard (QCO) for M570
IMS 70
1
Internal Siandjrd: THPFOS
Uie Area
Absolute Retention Time
Expected RT S:54
Current Mel hud
Noise Thtes s.d 5.0
Quant Thre
Mm. Wid S 5
Mult. W M 20 20
Bate. Widt UW too
RT Win. |ieo 20 20
1Smodi
1
SOO USOtOII t O U SOOUS O C O 1No Comment
ITHPfOS ute as Internal Standard
Expected RT 4:31
Current Method
Noise Thre 20.0 20.0
Quant Thre 4.01 4.0
Min. Wid 5 3
Mult. Wid 10 10
Base. Widt 100 100
RT Win. Iseo 25 25
Smoot1
1
t
Page 62
000660
Northwest Bioanalytical
Figure 22. Low Standard for PFOS
IffQS i Internai Standard: THPFOS
Uve Area
Absolute Retention Time
Expeaed RT 5:06
Current Method
Nose fh reQuant Hire
l.U
1.0
1.0
Min. Wfd Mult. Wid Base. WkJt
___ 5
20 00
5 20 100
RT Win. (secs 20 20
Smootl
1
S0M51001 1 OOt SOOU5 Stdl I I No Comment
Study No. NWBS00-I45 Report No. NWBRO1-155
ITHPfOS use ai Internai Standard
Eapeaed RT 4:31
Current Method
Noise Thres 20.a 20.a
Quant Thre 4.C 4.0
Min. Wid 5 s
Muli. Wid
10 10
Base. Wit 100 100
RT Win. Isecs 2S 2S
Smoot
11
S0 0U SQ I001 to o t SOO U SSuH 1 \ No Comment
Page 63
oooggi
Northwest Bioanalytical
Figure 23. Low Standard for PFOA
Internal Standard: fUPfOS Ute Arej Abtolute Retention Time
Curtent Method
Notte ihres 10.0 IU.0
Quant ihre 1.0 1.0
Min. Wid
S
8
Mull. Wid 16 16
Bate. W.dt 100 100
RT Win. (tee 26 26
Smooti 1 1
S0OU5O1001 1 00t SOOUSSidl > 1 No Comme
Study No. NWBS00-145 Report No. NWBROl-155
ITHPfOS ute Internal Standard
Expected RT 4:31
Current Method
Noite Threi 20.0 20.0
Qua Thre 4.C 4.0
Min. Wid S S
Mult, wid 10 10
Base. Wid 100 100
RT Win. (sect 25
25
Smoot 1 1
D0*ISOO\
1 SOOHSStl 1 1 No Comme
Page 64
000662
Northwest Bioanalytical
Figure 24. Low Standard for PFHS
IPFH5
1
Internal Standard: rU PfO S
Ute Area
Absolute Retention Time
Expeded RT 3:56
Current Method
Nrxse Thre s o i s .o
Quam Ihre
1.5 1.5
Min. Wid 5 5
Mult. Wid IB 18
Base. Wid! 100 to o
XT Win. isca 50 so
Smootl 1
$0014501001 I ooi 500145 Stdl t > No Comment
Study No. NWBS00-I45 Report No. NWBRO1-155
(THPfOS use at Internai Standard
Expected XT 4:31
Current Method
Noise Thres 20.0 20.0
Quant Thre 4.0 4.0
Min. Wid 5 5
Mult. Wid 10 10
Base. Wldt 100 to o
RT Win. (sea 25
25
Smoot 1 1
S001450100t 1 001 S00145 Sidt 1 i No Comment
Page 65
000663
Northwest Bioanalyticai
Figure 25. Low Standard for PFOSAA
Study No. NWBS00-I45 Report No. NWBRO1-155
iPfOSAA
1
Internoi Standard: THPF05
Use Arco
Absolute Retention Time Expected RT 6:03
Current Mediad
Noise Thret S,Q S.O
Quart Thre 1.3 1.3
Mm.Wid 6 6
Muh. Wkli 17 17
100Bose. Wldt 100
RT Win. (tea
20
Smootl_____ I
20
1
SOOUSOlOOl 1 001 SOOMS Sedi t I No Comment
ITHPfOS use as Internal Standard
Current Method
None Thre 2Q. 20.0
Quant Thre 4 0 4.0
Min. Wid 5 S
Mult. Wid
10 10
Base. With 100 100
RT Win. {sea 25 25
Smootl 1 I
SO0M 50100I I 001 $00145 Stdl 1 1 No Comment
Page 66
000664
Northwest Bioanalytical
Figure 26. Low Standard for PFOSA
Study No. NWBS00-I45 Report No. NWBR01-I55
IPfOSA
1
Internal Standard: THPFOS
Use Area
Absolute Retention Time
Expected RT 5:51
Current
Noise Ihre
7.C
Quant Thre 1.3
Min. Wid Mult. Wid
s 17
Base. Widt 100
RT Win. (sea
20
1
7.0 1.5
5 17 too 20
1
S0W 50I001 I 001 S0014S Sid 1 1 1 No Comment
Page 67
0OOGG5
Northwest Bioanalytical
Figure 27. Low Standard for M556
iM S S t
1
Internal Standard: THPFOS
Use Area
Absolute Retention Time
Expeaed RT 5:36
Current Method
Noise Thres . SO
Quant rhre 0.5 0.5
Min. Wid 5 5
MuH. Wid 21 U
Base. Wkil too 100
T Wm. (sec Smooil
20 20
11
SO0MS01002 1 002 SOOMS Std2 M N o Comme
Study No. NWBSOO-145 Report No. NWBRO1-155
Page 68
000666
Northwest Bioanalytical
Figure 28. Low Standard for M570
|m s ;o
I
Internai Standard: THPfOS
Ute Area
Absolute Retention Time
Expected R f S:54
Current Method
Norte Ihres 5.(
5.0
Quant Ihre
1.0 1.0
Min. Wld 5 S
Mull. Wid 20 20
Bate. Wldt 100 100
T Win. (tea 20 20
1!
Thu. N ov B. 2001 8:25 PM 8:59 in 1 period
1: 8:59 MRM. 642
570.0->4l9.0
A IM 6075
Height___ 1132____________
Start Time
5:38
End Time
6:02
0:24.4
eternimi Time
5:54 _
Integration Type A -B8
Study No. NWBS00-145 Report No. NWBR01-155
iTHPfOS
~I
use as Internal Standard
Expected RT 4:31
Current Method
Norte Thres 3 0 . 0 2 0 . 0
Quant Ihre 4.C 4.0
Min. Wid 5 5
Mult. Wid 1 0 t o
Bate, widi t o o t o o
RT Win. (secs
25
25
Smoot
1
1 11S0014501001 001 SOOUSStdl No 100i ;Thu. Nov 8. 2001 8:25 PM
0:59 in 1 period I: 8:59 MRM . 642 team
II.-
:^ : : , 52T
427.0*>407.0
Area 836307 Heicht 117889 Start Time End Time Integration Width Retention Time Integration Type
4:24 4:53 0:29.4 4:35 A - 68
"i
nUraity:.! 17938 cp*
. '::.
:.l -.
36 K 126 187 232 278
428 301 342 604
61 n iq> 241 301 361 421 9i >54i1 w60ji1 sSccmm*n--
051 42 232 3:23 4:13 3.01 5.54 6 M 7 33 323 Thw{*
Page 69
000667
Northwest Bioanalytical
Figure 29. High Standard for PFOS
jPt-OS ~~~
j
Internal Standard: HPFOS
Um Aiea
Absolute Retention Time
Expected RT 5:06
Current Method 4.0
50014501009 I 009 S00I45 Std9 I I N o Comment
Study No. NWBS00-I45 Report No. NWBRO1-155
Page 70
O fcO SSS
Northwest Bioanalytical
Figure 30. High Standard for PFOA
(___ 1
Internal Standard: TMPFOS Use Area Absolute Retention Time Expeded RT 4:3/
Current Method
Noise fhre: 10 0 t o o
Quant Ihre 3.01 3.0
Min. WkJ 8 8
Mull. Wld
16 16
Base. Widt t o o 100
RT Win. (sea
26
Smootl 1 1
SO14SO 1009 1 0O9SO0I45 Sd9 I l No
Study No. NWBS00-145 Report No. NWBR01-I55
{THPfOS
u m at Internai Standard
Expected KT 4:31
Current M ethod
Noise Ihre; 20.0 20-0
3uani Thre
4.0
Min. Wld
S
Mull. Wtd
10
Oase. Widt 100 100
' Win. (sec
25
Smoot!
S00I45 Std9 11 N a (
&.:'Thu. Nov 8. 2001 9:55 PM 8:59 >n I period
10 1
1: 8:59 M RM , 642 scans 427.fr >407.0
Area 702935 Beitritt 101201 Start Time End Time Interation Width Retention Time Inteeration Type
4:26 4:52 0:26.9 4:36 A-BB
BHEBi m
.90!
SO T' SO SO4030-
`.t M :
.-'- -
*iKSp#
^!:vi-
. >: '! Li ^4
: 4 :- - ;-
20-
IQ
S B
580 a t
I 121 1SI 241 Ml Ml 421 ' 481 541 SOI SmivOil 1:42 232 iffl 4:13 5 03 554 S:44 733 ' 3:23 Irrlj
Page 71
000669
Northwest Bioanalytical
Figure 31. High Standard for PFHS
F h s -------------------------- 1
Curren
u [> K C Id *T 3:5*
___ h l
M.n. V M
S
M.W W l RT Win. (sea
100 50
_____ 1
SOOMSO1O09 1 009 $OQU5$d9 t \ No Comment
I.} S 18
1
Study No. NWBS00-145 Report No. NWBR01-155
Page 72
000G70
Northwest Bioanalytieal
Figure 32. High Standard for PFOSAA
Study No. NWBSOO-145 Report No. NWBR01-155
iPfOSAA
1
Internal Standard: THPFOS
Use Area
Absolute Retention firne
Expeaed RT 6:03
Current Method Ni*se Thre 5.C 5.0 Quant Thre 1.J 1.3
Min, Wid 6 6 Mult. Wid 17 17 Base. Widi 100 t o o RT Win. (secs 20 70
Smooii 1 1
S0014501009 1 009S00145 Sid9 1 1 No Comment
1THPFS
~\
use es Internal Standard
Expected RT 4;J1
Current Method
Norse Thres 20.0 30.0
Quant Thre 4.C 4.0
Min. Wld 3 5
Mult. Wid t o 10
Base. Widt 100 100
RT Win. {secs 25
25
Smootl 1 1
S014501009 1 009S0014SSid9 1 I No Comment
Page 73
000671
Northwest Bioanalyttcal
Figure 33. High Standard for PFOSA
Study No. NWBS00-I45 Report No. NWBRO1-155
Ip f o s a
I
interna) Standard: FHPFOS
Use Area
Absolute Retention Time
Expected RT 5:51
Current Method
Noise Huet 7.0 7.0
Quant Hire I.S
1.5
Min. WkJ
5
5
MuJl. Wld 17 17
Base. Wich 100 too
RT Win. Isen
20
20
Smooil 1 1
S0014501009 1 009 $0014$ Std9 1 1 No Comment Thu, Nov 0. 2001 9:55 PM 0:59 in period
1: 0:59 MRM . 642 scans
497.9*>77.9
Area 225511) Heiehr 237946 Start Time End Time Intenration Width Retention Time Inle*ration Type
5:24 6:54 1:29 5:51 A B8
intmtty: 237946 ept
19 30 i : 150 l84r229'--`:2S5:'-': 342--
6i ; m $81 i 2*i ;30i ;r-zs.T
:H0J1 -1:42 . 232:;,
ITHPFOS utc as Interna) Standard
Expected RT 4:31
Current Method
Noise Hues 20.0 20.0
Quant Threi 4.0 4.0
Min. WkJ
5
5
Muil. Wid 10 10
Base. Widt 100 ICO
RT Win. (sea 25
25
Smoot 1 1
S0014501009 1 009 $00145 Sid9 1 I No Comment
^ 3 2 r 563 623: 481 n;341 VWl Sea
Page 74
000672
Northwest Bioanalytical
Figure 34. High Standard for M556
|Mi> 1
Internat Starnarti: THPFOS U m Area Absolute Retention Time Expected RT 5:36
Current Method
Noise Threi
5.0
Quant Thre 0.5 0.5
Min. Wid 5 S
Muli. Widi 2t 21 100Base. Wrdt IDO
RT Win. (sea 20 20
Smooil
I
S0014501009 I 009 500t 45 Sld9 1 I N o Comment
Study No. NWBS00-145 Report No. NWBR01-155
ITHPfOS use as Internal Standard
Expeed RT 4:3 t
Current Method
Noise Ihres
20.0
Quant Thre Min. Wid Mult. Widi I
4.0
5 10
Base. Widi _
100
RT Win. (se a ,
25
Smootl
S0014501009 I 0O 9S00I4S Std9 1 I No Comment
Page 75
000673
Northwest Bioanalytical
Figure 35. High Standard for M570
Study No. NWBSQO-145 Report No. NWBRO1-155
IMS 70
1
Internai Standard: fHPfOS
Ute Area
Abtoiute Retention Time
Expected RT 5:54
Current Method
N o i Threi
5.0
Quent (Tire' Min. Wid Mult. Wid Bate. Wldt
1.0 1.0
5s 20 20 100 100
R TW irui
S0OU501OO9 1 009 S00145 Std9 1 I No Comment Thu. N ova. 2001 9:55 PM
8:59 in 1 period
1: 0:59 MRM. 642 scant
570.0->419.0
Area 2674524 Heieht 379007 Start Time End Time Inteeration Width Retention Time Inteeration Tvoe
BSfflS
5:34 6:59 t:25 5:54 A-BV
:.;r.ati,.':-.y.?.' ; v r ~ ;
Mmiiy:3790l2epi
31 65 ..il
61
122 158 205 247 294
i l . 181 2< r so.-' :2.32 2 5 4:13- - : l . 5 3 4 - s : :
735 - 3:237 T M *
ITHPfQS ute as Internai Standard
Expected RT 4:31
Current Method
Notte Threj 20.0 20.0
Quant Thre 4.0l 4.0
Min. Wid 5 5
Mult. Wid 10 to
Bate. Widt 100 100
RT Win. (tea 25 2V
Smoul
1
S0OI450I0O9 1 009 S00145Std9
Thu. Nov 8. 2001 9:55 PM 8:59 in Y period
ICO-
1: 8:59 MRM . 642 scant
90
427.0->407.0
Area 702935
Heieht 101201
Slart Time
4:26
End Time
4:52
inteeration Width 0:26.9
Retention Time
4:36
Inteeration Tvoe A - B B
ItliHH Kgj
8-
f-W- : !v
9
h -;-; :.;7 71.
iSM iil.="::;+!-S
htmsrty: 101283 eps
11:
'-l"i .. ^ r
W im
155 201234
429 466 307 580 626
'61 ' 121 31 24t 301 6 I ' 42* 1 ' '5-li ' ' I ' Scorr a s i 1:42 2:32 323 4:13 5lOi 5:54 6:44 7 35 823 Timi
Page 76
000674
