Document X7Q7QQEpZ94nMOeD57ERzDegK
AR226-1740
STUDY TITLE Extraction of Potassium Perfluorooctanesulfonate from Egg Membrane, Albumen, and
Yolk for Analysis Using HPLC-Electrospray/Mass Spectrometry
DATA REQUIREMENTS Analytical Method Requirements
STUDY DIRECTOR Sean Gallagher
STUDY COMPLETED ON September 06,2001
PERFORMING LABORATORY / TESTING FACILITY Centre Analytical Laboratories, Inc. (Centre) 3048 Research Drive State College, PA 16801
' Phone: 814-231-8032 STUDY SPONSOR
3M Environmental Technology and Safety Services Building 2-3E-09 PO Box 33331
St. Paul, MN 55133-3331 PROJECT
Centre Protocol Number: 00P-023-063 Centre Study Number: 023-063 Total Pages: 102
Centre Study No.: 023*063
GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT
Centre Study Number 023-063 entitled "Extraction o f Potassium Perfluorooctanesulfonate from Egg Membrane, Albumen, and Yolk for Analysis Using HPLC-Eleccrospray/Mass Spectrometry," conducted for 3M Environmental Technology and Safety Services, was performed in compliance with US EPA TSCA G ood Laboratory Practice Standards (40 CFR Part 792) by Centre Analytical Laboratories, Inc with the follow ing exceptions. 1. 792.63. M aintenance and Calibration of Equipment. N o equipment SOP was in
place for the bead beater used in the extraction at the tune o f use.
W. ;ker
. esdgator
Centre Analytical Laboratories, Inc.
Sean vauiagner Study Director W ildlife International, Ltd.
Centre Analytical Laboratories, Inc.
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QUALITY ASSURANCE STATEM ENT
Centre Analytical Laboratories' Quality Assurance Unit reviewed Centre Study Number 023-063, entitled, "Extraction of Potassium Perfluorooctanesulfonate from Egg Membrane, Albumen, and Yolk for Analysis Using HPLC-Electrospray/Mass Spectrometry". All examined phases were reviewed for conduct according to Centre Analytical Laboratories' Standard Operating Procedures, the Study Protocol, and all applicable Good Laboratory Practice Standards. All findings were reported to the Study Director and to management.
Phase
Date Inspected
Date Reported to Date Reported to
Centre
Study Director and
Management Sponsor Management
1. Protocol Review
3/30/01
4/9/01
5/7/01
2. Extraction, Fortification
3/30/01
4/9/01
5/7/01
3. Raw Data Review 4/5-9/01
4/18/01
5/7/01
4. Raw Data and Draft Report Review
4/17-18/01
4/26/01
5/7/01
5. Final Report Review
6/28/01
9/6/01
9/6/01
Date
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Centre Study No.; 023-063
CERTIFICATION OF AUTHENTICITY
This report, for Centre Study Number 023-063. is a tree and com plete reprsentation of the raw data for the study.
Submitted by.
Centre Analytical Laboratories, Inc. 3048 Research Drive State C ollege, PA 16801 (814)231-8032
Principal Investigator, Centre;
frTWjueC
jD eck er
Scientist Centre Analytical Laboratories, Inc.
Date
Centre Analytical Laboratories, Inc, Facility Management;
Richard A. ' President Centre Analytical Laboratories, Inc.
cj-it- l D ate
<?-jW-ol
Study Director
iL u ^ Sean Gallagher W ildlife International, Ltd.
Sponsor Representative
Centre Analytical Laboratories, Inc.
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IS S S *1
Centre Study No.: 023-063
STUDY IDENTIFICATION
"Extraction of Potassium Perfluorooctanesulfonate from Egg Membrane, Albumen, and Yolk for Analysis Using HPLC-Electrospray/Mass Spectrometry,"
CENTRE PROTOCOL NUMBER: 00P-023-063
CENTRE STUDY NUMBER:
023-063
TYPE OF STUDY:
Analytical
SAMPLE MATRIX:
Mallard and Quail Egg Membrane, Albumen, and Yolk
TEST SUBSTANCE:
Perfluorooctanesulfonate (PFOS)
SPONSOR:
3M Environmental Technology and Safety Services Building 2-3E-09 PO Box 33331 St. Paul, MN 55133-3331
STUDY DIRECTOR:
Sean Gallagher
W ildlife International, Ltd.
8598 Commerce Drive Easton, MD 21601
TESTING FACILITY:
Centre Analytical Laboratories, Inc. 3048 Research Drive State College, PA 16801
ANALYTICAL PHASE TIMETABLE:
Study Initiation Date: Experimental Start Date: Experimental Termination Date: Study Completion Date:
03/26/01 03/26/01 04/02/01 09/06/01
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PRO TECT PERSO NNEL
The Study Director for this project was Sean Gallagher at Wildlife International, Ltd. The following personnel from Centre Analytical Laboratories, Inc., were associated with various phases of the study:
Name Emily Decker Dave Bell Mitra Atjmand Ling Ling Liu Yu Qiong Tan Xiaoming Zhu Lawrence Ord Rickey Keller Shawn Robb
Title Scientist Scientist Technician Technician Technician Technician Sample Custodian Sample Custodian Sample Custodian
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TABLE OF CONTENTS
Page TITLE PAGE...................................................................................................................... 1 GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT............................. 2 QUALITY ASSURANCE STATEMENT.......................................................................... 3 CERTIFICATION OF AUTHENTICITY........................................................................... 4 STUDY IDENTIFICATION............................................................................................... 5 PROJECT PERSONNEL.................................................................................................... 6 TABLE OF CONTENTS.................................................................................................... 7 LIST OF TABLES.............................................................................................................. 8 LIST OF FIGURES........................................................................................................... 11 LIST OF APPENDICES................................................................................................... 12 1.0 SUMMARY............................................................................................................... 13 2.0 OBJECTIVE............................................................................................................... 13 3.0 INTRODUCTION...................................................................................................... 13 4.0 TEST SYSTEM......................................................................................................... 14 5.0 REFERENCE MATERIAL....................................................................................... 14 6.0 DESCRIPTION OF ANALYTICAL METHOD........................................................15
6.1 Extraction Procedure................................................................................................ 15 6.2 Preparation of Standards and Fortification Solutions.............................................. 15 6.3 Chromatography....................................................................................................... 16 6.4 Instrument Sensitivity.............................................................................................. 16 6.5 Description of Instrument and Operating Conditions...............................................17 6.6Quantitation and Example Calculation.................................................................... 18 7.0 EXPERIMENTAL DESIGN...................................................................................... 19 8.0 RESULTS................................................................................................................... 20 9.0 CONCLUSIONS..................................................................................................... 20 10.0 RETENTION OF DATA AND SAMPLES............................................................. 21
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Table I.
LIST OF TABLES Page
Summary of PFOS in Reagent Blanks........................................................... 23
Table EL Summary of PFOS in Mallard Membrane Blanks......................................... 24
Table HI. Summary of PFOS in Mallard Albumen Blanks............................................ 24
Table IV. Summary of PFOS in Mallard Yolk Blanks.................................................. 24
Table V. Summary of PFOS Recoveries in Mallard Membrane................................... 25
Table VI. Summary of PFOS Recoveries in Mallard Albumen..................................... 25
Table VII. Summary of PFOS Recoveries in Mallard Yolk............................................ 26
Table VUI. Summary of PFOS for Week 1 Mallard Membrane Control Samples.......... 26
Table IX. Summary of PFOS for Week 1 Mallard Membrane Samples at 6.2 ppm ai.....................................................................................................27
Table X. Summary of PFOS for Week 1 Mallard Membrane Samples at 17.6 ppm ai..................................................................................................... 27
Table XI. Summary of PFOS for Week 1 Mallard Albumen Control Samples............ 28
Table XH. Summary of PFOS for Week 1 Mallard Albumen Samples at 6.2 ppm a i.... 28
Table XDI. Summary of PFOS for Week 1 Mallard Albumen Samples at 17.6 ppm a i.. 29
Table XIV. Summary of PFOS for Week 1 Mallard Yolk Control Samples.................... 29
Table XV. Summary of PFOS for Week 1 Mallard Yolk Samples at 6.2 ppm a i........... 30
Table XVI. Summary of PFOS for Week 1 Mallard Yolk Samples at 17.6 ppm a i......... 30
Table XVII. Summary of PFOS for Week 6Mallard Membrane Control Samples.........31
Table XVIII. Summary of PFOS for Week 6Mallard Membrane Samples at 6.2 ppm ai...................................................................................................31
Table XIX. Summary of PFOS for Week 6Mallard Membrane Samples at 17.6 ppm ai..................................................................................................... 32
Table XX. Summary of PFOS for Week 6Mallard Albumen Control Samples............ 32
Table XXI. Summary of PFOS for Week 6Mallard Albumen Samples at 6.2 ppm a i.... 33
Table XXH. Summary of PFOS for Week 6Mallard Albumen Samples at 17.6 ppm a i. 33
Table XXIII. Summary of PFOS for Week 6Mallard Yolk Control Samples........... 34
Table XXIV. Summary of PFOS for Week 6Mallard Yolk Samples at 6.2 ppm a i .. 34
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Table XXV. Summary of PFOS for Week 6Mallard Yolk Samples at 17.6 ppm a i .... 35
Table XXVI. Summary of PFOS in Quail Membrane Blanks...................................35
Table XXVII. Summary of PFOS in Quail Albumen Blanks..........................................35
Table XXVIII. Summary of PFOS in Quail Yolk Blanks................................................36
Table XXIX. Summary of PFOS Recoveries in Quail Membrane................................... 36
Table XXX. Summary of PFOS Recoveries in Quail Albumen...................................... 37
Table XXXI. Summary of PFOS Recoveries in Quail Yolk............................................ 37
Table XXXII. Summary of PFOS for Week 1 Quail Membrane Control Samples......... 38
Table XXXIII. Summary of PFOS for Week 1 Quail Membrane Samples at 6.2 ppm ai......................................................
38
Table XXXIV. Summary of PFOS for Week 1 Quail Membrane Samples at 17.6 ppm ai..............................................................................................39
Table XXXV. Summary of PFOS for Week 1 Quail Albumen Control Samples........... 39
Table XXXVI. Summary of PFOS for Week 1 Quail Albumen Samples at 6.2 ppm ai... 40
Table XXXVII. Summary of PFOS for Week 1 Quail Albumen Samples at 17.6 ppm a i............................................................................................40
Table XXXVUI. Summary of PFOS for Week 1 Quail Yolk Control Samples............... 41
Table XXXIX. Summary of PFOS for Week 1 Quail Yolk Samples at 6.2 ppm a i........ 41
Table XL. Summary of PFOS for Week 1 Quail Yolk Samples at 17.6 ppm ai.............42
Table XLL Summary of PFOS for Week 6 Quail Membrane Control Samples............... 42
Table XLII. Summary of PFOS for Week 6Quail Membrane Samples at 6.2 ppm a i.....43
Table XLIII. Summary of PFOS for Week 6Quail Membrane Samples at 17.6 ppm ai.. 43
Table XLTV. Summary of PFOS for Week 6 Quail Albumen Control Samples.............. 44
Table XLV. Summary of PFOS for Week 6 Quail Albumen Samples at 6.2 ppm a i...... 44
Table XLVI. Summary of PFOS for Week 6Quail Albumen Samples at 17.6 ppm ai.... 45
Table XLVII. Summary of PFOS for Week 6 Quail Yolk Control Samples................... 45
Table XLVIIL Summary of PFOS for Week 6Quail Yolk Samples at 6.2 ppm a i......... 46
Table DL. Summary of PFOS for Week 6Quail Yolk Samples at 17.6 ppm ai.............. 46
Table L. Summary of Moisture Determinations for Mallard Membrane Samples.......47
Table LI. Summary of Moisture Determinations for Mallard Albumen Samples......... 48
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Table LH Summary of Moisture Determinations for Mallard Yolk Samples................49 Table LEI. Summary of Moisture Determinations for Quail Membrane Samples.......... 50 Table LTV. Summary of Moisture Determinations for Quail Albumen Samples............. 51 Table LV. Summary of Moisture Determinations for Quail Yoik Samples.................... 52
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LIST OF FIGURES Page
Figure 1. Typical Calibration Curve for PFOS............................................................... 54
Figure 2. Chromatogram Representing a 0.1 ng/mL standard for PFOS....................... 55 Figure 3. Chromatogram Representing a 1.0 ng/mL standard for PFOS....................... 56 Figure 4. Chromatogram Representing a Reagent Blank for PFOS
(Centre ID Reagent Blank A, Set: 032601A)................................................ 57
Figure 5. Chromatogram Representing Control Mallard Membrane for PFOS (Centre ID: 0003802 Blank A, Set: 032601C)........................................................... 58
Figure 6. Chromatogram Representing Control Mallard Membrane Fortified with 10 ppb of PFOS, (Centre ID: 0003802 Spk A Dup. Inj., Set: 032601C)........... 59
Figure 7. Chromatogram Representing Mallard Membrane Sample from Week 1 at 6.2 ppm ai (E01-0263-20825) (Centre ID: 0103092, Set: 032601C)........ 60
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LIST OF APPENDICES
Page
Appendix A Study Protocol 00P-023-063 (Centre Study No. 023-063) and Amendments and Deviation..................................................................... 61
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1.0 SU M M ARY
Centre Analytical Laboratories, Inc. (Centre) extracted mallard and quail egg membrane, albumen and yolk samples for the determination of perfluorooctanesulfonate (PFOS) according to protocol 00P-023-063 (Appendix A).
The limit of quantitation for each matrix was 10 ppb. The LOQ for each matrix was determined in a method validation study performed at Centre (Centre Study: 023-027). PFOS ranged from non-quantifiable (where non-quantifiable is defined as less than the concentration of the lowest calibration standard (0.1 ng/mL)) levels to 434 ng/g in the mallard membrane samples, from non-quantifiable levels to 3550 ng/g in the mallard albumen samples, and from non-quantifiable levels to 63,500 ng/g in the mallard yolk samples. PFOS ranged from non-quantifiable levels to 20,900 ng/g in the quail membrane samples, from non-quantifiable levels to 93.9 ng/g in the quail albumen samples, and from non-quantifiable levels to 65,500 ng/g in the quail yolk samples. All of these results are reported on a wet weight basis.
The average percent recoveries standard deviations for PFOS in mallard membrane, albumen, and yolk samples were 91% 6%, 91% 16%, and 93% 13%, respectively. The average percent recoveries standard deviations for PFOS in quail membrane, albumen, and yolk samples were 102% 11%, 85% 5%, and 96% 7%, respectively.
2.0 O BJECTIVE
The objective of this analytical phase of the study was to determine levels of perfluorooctanesulfonate (PFOS) in specimens of mallard and quail egg membrane, albumen, and yolk using the analytical method titled, "Determination of Perfluorooctanesulfonate in Egg Membrane, Ablumen and Yolk by LC/MS/MS".
3.0 IN TRO DUC TIO N
This report details the results of the analysis for the determination of PFOS in mallard and quail egg membrane, albumen, and yolk using the analytical method entitled, "Determination of Perfluorooctanesulfonate in Egg Membrane, Ablumen and Yolk by LC/MS/MS".
This analytical portion of the study was initiated on March 26, 2001, when the study director signed Centre protocol number 00P-023-063. The experimental start date was March 26,2001, and the experimental termination date was April 2,2001.
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4.0 TEST SYSTEM
The control mallard and quail egg membrane, albumen, and yolk used for the matrix
blanks and matrix fortifications were received on May 18, 2000 from Wildlife
International Ltd., Easton, MD and stored frozen at Centre. The following Centre ID's
were assigned to the different matrices:
Matrix
Centre ID
Mallard Membrane
0003802
Mallard Albumen
0003800
Mallard Yolk
0003799
Quail Membrane
0003798
Quail Albumen
0003796
Quail Yolk
0003795
On February 23, 2001, 308 quail egg samples were received. The samples were stored frozen until they were logged in by Centre personnel on February 26, 2001 and stored frozen. On February 24, 2001, 194 mallard egg samples were received. The samples were stored frozen until they were logged in by Centre personnel on March 1, 2001 and stored frozen. Each egg sample received was divided into membrane, albumen, yolk, and shell. Each component of the egg was given a unique Centre ID at the time of log-in. Although all of the egg samples were received at Centre, only those designated by the sponsor were extracted and analyzed.
Sample log-in and chain of custody information can be found in the raw data package associated with this study. Storage records will be kept at Centre Analytical Laboratories, Inc. and a true copy of the storage records can be found in the raw data package associated with this study.
5.0 REFERENCE M ATERIAL
The analytical standard PFOS was received at Centre on June 3, 2000 from 3M Environmental Technology and Services.
Stability was not assessed in this study but the reference material was stored according to the certificate of analysis (frozen) and was used prior to expiration. The solubility of the reference material was not relevant to this study. The available information for the reference material is listed below.
Compound PFOS
Centre Control No.. TCR No. Purity (%) Expiration Date
00-023-042
TCR 00017-46 97.9
08/31/01
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The molecular structure of PFOS is given below.
PFOS Chemical Name Molecular weight
O
= =
Perfluorooctanesulfonate 499 (CgFnSCV)
C8F17S-- O'
o
Note: The neutral molecule and standard from which the PFOS (anion) is obtained, is potassium perfluorooctanesulfonate [C8F17SO3K], molecular weight 538.
6.0 D ESC R IPTIO N O F ANALYTICAL M ETH O D
Analytical method entitled "Determination of Perfluorooctanesulfonate in Egg Membrane, Ablumen and Yolk by LC/MS/MS" was used for this study.
6.1 Extraction Procedure
Egg membrane
A 0.1 g sample was used for the extraction procedure. One milliliter of methanol was added to the sample along with three stainless steel beads. The sample was then placed on a bead beater set at position 7 for 5 minutes. The sample was then centrifuged on a mini-centrifuge at ~ 2000 rpm for ~ 2 minutes. Each sample was filtered and analyzed by LC/MS/MS electrospray.
Egg albumen and yolk
A 1.0 g sample was used for the extraction procedure. Twenty-five milliliters of methanol were added to the sample. The sample was placed on a wrist-action shaker for 15 minutes and then centrifuged at ~ 2000 rpm for ~ 10 minutes. A five milliliter aliquot of the sample was taken and 0.5 g of carbon was added. The sample was shaken then allowed to sit. Each sample was filtered and analyzed by LC/MS/MS electrospray.
6.2 Preparation of Standards and Fortification Solutions
Standard solutions were prepared on March 14, 2001 as specified in Centre Analytical Laboratories' protocol 00P-023-027. An individual stock standard solution of PFOS was prepared at a concentration of 100 pg/mL by dissolving 10 mg of the standard (corrected for purity and salt content) in methanol. From this solution, a 1.0 pg/mL fortification standard solution was prepared by taking 1 mL of the stock and bringing the volume up to 100 mL with methanol.
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The 0.1 |ig/mL fortification standard was prepared by taking 10 mL of the 1.0 pg/mL fortification standard and bringing the volume up to 100 mL with methanol. A 0.01 pg/mL standard was prepared by taking 10 mL of the 0.1 pg/mL standard and bringing to 100 mL with methanol.
A set of standards containing PFOS was prepared by dilution of the 0.1 pg/mL and 0.01 pg/mL solutions in the following manner.
Initial Cone. (pg/mL)1 0.1 0.1 0.1 0.01 0.01 0.01
1of PFOS
Volume (mL) 5 2 1 5 2 1
Diluted to (mL) 100 100 100 100 100 100
Final Cone. (pg/mL) 0.005 0.002 0.001 0.0005 0.0002 0.0001
The stock standard solution and all fortification and calibration standard solutions were stored in a refrigerator (4C 2C) when not in use. Documentation of standard preparation can be found in the raw data associated with this report.
6.3 Chromatography
Quantification of PFOS was accomplished by LC/MS/MS electrospray. The retention time of PFOS was ~ 4.4 min. Peaks were detected in the control matrices corresponding to the analyte retention time, but the amounts detected were only significant enough to
alter the results in several o f the recoveries reported and the rest were less than the lowest
calibration standard (0.0001 pg/mL).
6.4 Instrument Sensitivity
The smallest standard amount injected during the chromatographic run had a concentration of 0.0001 pg/mL of PFOS.
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6.5 Description of Instrument and Operating Conditions Instrument: PE SCIEX API 3000 Biomolecular Mass Analyzer
Interface: SCIEX Turboion Spray Liquid Introduction Interface
Computer: Dell UltraScan PI 110
Software: HPLC:
PE SciexAnalyst version 1.1 Windows NT, version 4 Hewlett Packard (HP) Series 1100
HP Quat Pump HP Vacuum Degasser HP Autosampler HP Column Oven
HPLC Column:Genesis Cg (Jones Chromatography), 2.1 mm x 50 mm, 4jx Column Temp.: 35 C Injection Vol.: 10 pL Mobile Phase (A): 2 mM Ammonium Acetate in ASTM type I water Mobile Phase (B): Methanol Flow Rate: 0.3 mlVmin.
Time % A % B
0 60 40
1.0 0 100
7.0 0 100
7.5 60 40
11.0 60
40
Ions monitored:
Analvte PFOS
Mode negative
Transition Monitored 499 99
Approximate Retention Time (min)
4.40
Tune File Parameters
Controls IS-Iospray DP-Declustering Potential FP-Focusing Potential EP-Entrance Potential CE-Collision Energy CXP-Collision Cell Exit Potential
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Set -4200.0
-51.0 -230.0 10.0
-70 -5
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DF-Deflector CEM-Channel Electron Multiplier
300.0 2800.0
Gas Flows
Nebulizer Gas Curtain Gas Collision Gas TIS Temperature
Set
12 14 4 350C
6.6 Quantitation and Example Calculation
Ten microliters of sample or calibration standard were injected into the LC/MS/MS. The peak area was measured and the standard curve was generated (using 1/x weighted linear regression) by Analyst software using at least five concentrations of standards. The concentrations for mallard and quail egg membrane, albumen, and yolk (ng/g) were determined from the equations below.
Equation 1 calculated the amount of analyte found (in ng/mL, based on peak area) using the standard curve (linear regression parameters) generated by the Analyst software program. Then Equation 2 calculated the amount of analyte found in ng/g.
Equation 1:
Analyte found (ng/mL) = (Peak area - intercept) slope
Equation 2: Analyte found (ng/g) = anal, found (ng/mL) x FV (mL) x DF sample wt. (g)
Where: FV = Final Volume
DF = Dilution Factor
For samples fortified with known amounts of PFOS prior to extraction, Equation 3 calculated the percent recovery.
Equation 3:
Recovery (%) =
(anal. Found (ng/g) - avg. anal, in Ctrl (ng/g) x 100 amount added (ng/g)
An example of a calculation using an actual sample follows: Mallard albumen sample Centre ID 0003800 Spk A (Set: 032601A), fortified at 10 pbb with PFOS, where:
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peak area intercept slope dilution factor ng/g added (fort level) avg. amt in controls final volume sample weight
= = = = =
= = =
3369 971 6700 1 10ng/g
0 (Not quantifiable) 25 mL 1.0038 g
Note: Not quantifiable is defined as less than the concentration of the lowest standard (0.1 ng/mL).
From equation 1: Analyte found (ng/mL)
= T3369- 9711 6700
From equation 2: Analyte found (ng/g) =
0.36 ng/mL
(0.36 ne/mL x 25 mL x l ) 1.0038 (g)
From equation 3: % Recovery
ZZ 8.97 ng/mL
= (8.97 ne/e x - 0 ne/e) x 100 10ng/g
= 90% Note: This example calculation was done using rounded numbers, and therefore may be slightly different from the values shown in the RAW DATA.
7.0 EXPERIM ENTAL D ESIG N
Each set of samples consisted of one reagent blank, one matrix blank, two matrix blanks fortified at known concentrations, and ~ 20 samples. Each sample was extracted using the appropriate method and then analyzed in duplicate. Also, in each set, a Type I H2O blank was analyzed.
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8.0 RESULTS
PFOS found in the reagent blanks are listed in Table I. The PFOS in the mallard membrane, albumen, and yolk blanks are given in Tables II-IV. The PFOS in the quail membrane, albumen, and yolk blanks are given in Tables XXVI-XXVIII. PFOS was detected in several of the control matrices, however few were significant enough to alter the recoveries reported.
Individual recoveries for PFOS in the mallard membrane, albumen, and yolk samples are detailed in Tables V-VII. The average percent recoveries standard deviations for PFOS in mallard membrane, albumen, and yolk samples were 91% 6%, 91% 16%, and 93% 13%, respectively. Individual recoveries for PFOS in the quail membrane, albumen, and yolk samples are detailed in Table XXIX-XXX1. The average percent recoveries standard deviations for PFOS in quail membrane, albumen, and yolk samples were 102% 11%, 85% 5%, and 96% 7%, respectively.
PFOS ranged from non-quantifiable (where non-quantifiable is defined as less than the concentration of the lowest calibration standard (0.1 ng/mL)) levels to 434 ng/g in the mallard membrane samples, from non-quantifiable levels to 3550 ng/g in the mallard albumen samples, and from non-quantifiable levels to 63,500 ng/g in the mallard yolk samples. These results are reported on a wet weight basis. Individual results are listed in Tables VIII-XXV. Some of the high levels of PFOS reported for some of the mallard egg membrane and egg albumen samples could be from possible contamination from the egg yolk or outer shell.
PFOS ranged from non-quantifiable levels to 20,900 ng/g in the quail membrane samples, from non-quantifiable levels to 93.9 ng/g in the quail albumen samples, and from non quantifiable levels to 65,500 ng/g in the quail yolk samples. These results are reported on
a wet weight basis. Individual results are listed in T ables XXXII-IL. Some o f the high
levels of PFOS reported for some of the quail egg membrane and egg albumen samples could be from possible contamination from the egg yolk or outer shell. The Sponsor Representative also informed Centre that the quail outer shells were not washed prior to separation of the internal components.
The average percent moistures for the mallard and quail egg membrane, albumen, and yolk samples are given in Tables L-LV.
9.0 CO NCLUSIO NS
The mallard and quail egg membrane, albumen, and yolk samples were successfully extracted and analyzed according to protocol 00P-023-063.
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10.0 R E T E N T IO N O F DATA A N D SA M PLES
When the final report is complete, all original paper data generated by Centre Analytical Laboratories, Inc. will be shipped to the sponsor. This does not include facility-specific raw data such as instrument logs, however exact copies of temperature logs will be submitted. Exact copies of all raw data, as well as a signed copy of the final analytical report and all original facility-specific raw data, will be retained in the Centre Analytical Laboratories, Inc. archives for the period of time specified in 40 CFR Part 792. Retained samples of reference substances are archived by the sponsor.
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TABLES
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Table I. Summary of PFOS in Reagent Blanks
Sponsor ID
na na na na na na na na na na na na na na na na na na na na na na na na na na
Centre ID
Reagent Blank A Reagent Blank A* Reagent Blank A Reagent Blank A* Reagent Blank A Reagent Blank A* Reagent Blank A Reagent Blank A* Reagent Blank A Reagent Blank A* Reagent Blank A Reagent Blank A* Reagent Blank A Reagent Blank A* Reagent Blank A Reagent Blank A* Reagent Blank A Reagent Blank A* Reagent Blank A Reagent Blank A* Reagent Blank A Reagent Blank A* Reagent Blank A Reagent Blank A* Reagent Blank A Reagent Blank A*
* Duplicate injection
Set Number
032601A 032601A 032601B 032601B 032601C 032601C 032701A 032701A 032701B 032701B 032701C 032701C 032801A 032801A 032801B 032801B 032901A 032901A 032901B 032901B 033001A 033001A 033001B 03300IB 033001C 033001C
Extraction Analysis
Date Date
3/26/01 3/26/01 3/26/01 3/26/01 3/26/01 3/26/01 3/27/01 3/27/01 3/27/01 3/27/01 3/27/01 3/27/01 3/28/01 3/28/01 3/28/01 3/28/01 3/29/01 3/29/01 3/29/01
3/26-27/01 3/26-27/01
3/27/01 3/27/01 3/27/01 3/27/01 3/28/01 3/28/01 3/28-29/01 3/28-29/01 3/29/01 3/29/01 3/30/01 3/30/01 3/30/01 3/30/01 3/31/01 3/31/01 3/31-4/1/01
3/29/01 3/31-4/1/01
3/30/01
4/1/01
3/30/01
4/1/01
3/30/01
4/2/01
3/30/01
4/2/01
3/30/01
4/2/01
3/30/01
4/2/01
AVERAGE:
STANDARD DEVIATION:
Analyte Found (ppb)
NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ
NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) LOQ = 10 ppb
Centre A nalytical Laboratories, Inc.
Page 23 o f 102
Centre Study N o.: 023-063
Table II. Summary of PFOS in Mallard Membrane Blanks
Sponsor ID
na na na na
Centre ID
0003802 Blank A 0003802 Blank A* 0003802 Blank A 0003802 Blank A*
Set Number
032601C 032601C 032701C 032701C
Extraction Analysis
Date Date
3/26/01
3/27/01
3/26/01 3/27/01
3/27/01 3/29/01
3/27/01
3/29/01
AVERAGE:
STANDARD DEVIATION:
Analyte Found (ppb)
NQ NQ NQ NQ NQ NQ
Table III. Summary of PFOS in Mallard Albumen Blanks
Sponsor ID
na na na na na na
Centre ID
0003800 Blank A 0003800 Blank A* 0003800 Blank A 0003800 Blank A* 0003800 Blank A 0003800 Blank A*
Set Number
032601A 032601A 032701B 032701B 033001C 033001C
Extraction Analysis
Date Date
3/26/01 3/26/01 3/27/01 3/27/01 3/30/01 3/30/01
3/26-27/01 3/26-27/01 3/28-29/01 3/28-29/01
4/2/01 4/2/01 AVERAGE:
STANDARD DEVIATION:
Analyte Found (ppb)
NQ NQ NQ NQ 6.03 6.59 2.94 2.62
Table IV. Summary of PFOS in Mallard Yolk Blanks
Sponsor ID
na na na na
Centre ID
0003799 Blank A 0003799 Blank A* 0003799 Blank A 0003799 Blank A*
Set Number
032601B 032601B 032701A 032701A
Extraction Analysis Date Date
3/26/01
3/27/01
3/26/01
3/27/01
3/27/01
3/28/01
3/27/01
3/28/01
AVERAGE:
STANDARD DEVIATION:
Analyte Found (ppb)
NQ NQ 9.57 9.80 5.48 4.87
* Duplicate injection
NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) LOQ = 10 ppb
For residues recorded as NQ, half of the concentration of the lowest standard (1.25 ppb) was used to calculate the average and standard deviation.
Centre A nalytical Laboratories, Inc.
Page 24 o f 102
Centre Study N o.: 023-063
Table V. Summary of PFOS Recoveries in Mallard Membrane
Sponsor ID
na na na na na na na na
Centre ID
0003802 Spk A 0003802 Spk A* 0003802 Spk B 0003802 SpkB* 0003802 Spk A 0003802 Spk A* 0003802 Spk B 0003802 SpkB*
Set Number
032601C 032601C 032601C 032601C 032701C 032701C 032701C 032701C
Extraction Analysis
Amt
Date Date Added (ppb)
3/26/01 3/26/01
3/27/01 3/27/01
10 10
3/26/01 3/26/01
3/27/01 3/27/01
100 100
3/27/01 3/27/01
3/29/01 3/29/01
10 10
3/27/01
3/29/01
97
3/27/01
3/29/01
97
AVERAGE:
STANDARD DEVIATION:
RELATIVE STANDARD DEVIATION:
%
Recovery
86 87 87 81 97 94 95 98 91 6 7
Table VI. Summary of PFOS Recoveries in Mallard Albumen
Sponsor ID
na na na na na na na na na na na na
Centre
ID
0003800 Spk A 0003800 Spk A* 0003800 Spk B 0003800 SpkB* 0003800 Spk A 0003800 Spk A* 0003800 Spk B 0003800 SpkB* 0003800 Spk A 0003800 Spk A* 0003800 SpkB 0003800 SpkB*
Set
Number
032601A 032601A 032601A 032601A 032701B 03270IB 032701B 032701B 033001C 033001C 033001C 033001C
Extraction Analysis
Amt
Date Date Added (ppb)
3/26/01 3/26/01 3/26/01 3/26/01 3/27/01 3/27/01 3/27/01 3/27/01 3/30/01 3/30/01 3/30/01 3/30/01
3/26-27/01 3/26-27/01 3/26-27/01 3/26-27/01 3/28-29/01 3/28-29/01 3/28-29/01 3/28-29/01
4/2/01 4/2/01 4/2/01 4/2/01
10 10 99 99 10 10 99 99 10 10 98474 98474 AVERAGE:
STANDARD DEVIATION: RELATIVE STANDARD DEVIATION:
%
Recovery
89 91 94 92 107 105 101 99 59 61 99 99 91 16 17
* Duplicate Injection
LOQ = 10 ppb
Centre A nalytical Laboratories, Inc.
Page 25 o f 102
Centre Study N o.: 023-063
Table VII. Summary of PFOS Recoveries in Mallard Yolk
Sponsor ID
Na Na Na Na Na Na Na Na
Centre ID
0003799 Spk A 0003799 Spk A* 0003799 Spk B 0003799 SpkB* 0003799 Spk A 0003799 Spk A* 0003799 SpkB 0003799 SpkB*
Set Number
032601B 032601B 032601B 032601B 032701A 032701A 032701A 032701A
Extraction Analysis
Amt
Date Date Added (ppb)
3/26/01 3/26/01 3/26/01 3/26/01 3/27/01 3/27/01 3/27/01 3/27/01
3/27/01 3/27/01 3/27/01 3/27/01 3/28/01 3/28/01 3/28/01 3/28/01
10 10 98 98 10 10 97 97
AVERAGE: STANDARD DEVIATION: RELATIVE STANDARD DEVIATION:
% Recovery
104 104 99 99 71 73 98 94 93 13 14
Table VIII. Summary of PFOS for Week 1 Mallard Membrane Control Samples
Sponsor ID
E01-0263-20852 EOI-0263-20852* EOl-0263-20853 E01-0263-20853* E01-0263-20854 E01-0263-20854*
E01-0263-20842
E01-0263-20842* E01-0263-20843 EOI-0263-20843*
Centre ID
0103200 0103200 0103204 0103204 0103208 0103208
0103160
0103160 0103164 0103164
* Duplicate Injection
Set Number
032601C 032601C 032601C 032601C 032601C 032601C
032601C
032601C 032601C 032601C
Extraction Analysis
Date Date
3/26/01 3/26/01
3/27/01 3/27/01
3/26/01 3/26/01
3/27/01 3/27/01
3/26/01 3/26/01
3/26/01
3/27/01 3/27/01
3/27/01
3/26/01 3/26/01 3/26/01
3/27/01 3/27/01 3/27/01
AVERAGE:
STANDARD DEVIATION:
Analyte Found (ppb)
NQ NQ NQ NQ NQ NQ
NQ
NQ NQ NQ NQ NQ
NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) LOQ = 10 ppb
Centre A nalytical Laboratories, Inc.
Page 26 o f 102
Centre Study N o.: 023-063
Table IX. Summary of PFOS for Week 1 Mallard Membrane Samples at 6.2 ppm ai
Sponsor ID
E01-0263-20825 E01-0263-20825* E01-0263-20846 E01-0263-20846* E01-0263-20832 E01-0263-20832* EOI-0263-20847 EOl-0263-20847* E01-0263-20848 E01-0263-20848*
Centre ID
0103092 0103092 0103176 0103176 0103120 0103120 0103180 0103180 0103184 0103184
Set Number
032601C 032601C 032601C 032601C 032601C 032601C 032601C 032601C 032601C 032601C
Extraction Analysis
Date Date
3/26/01 3/26/01 3/26/01 3/26/01 3/26/01 3/26/01 3/26/01 3/26/01 3/26/01 3/26/01
3/27/01 3/27/01 3/27/01 3/27/01 3/27/01 3/27/01 3/27/01 3/27/01 3/27/01 3/27/01 AVERAGE:
STANDARD DEVIATION:
Analyte Found (ppb)
168** 163** 46.8 46.2 27.1 27.2 33.1 32.9 53.4 53.8 40.1 11.2
Table X. Summary of PFOS for Week 1 Mallard Membrane Samples at 17.6 ppm ai
Sponsor ID
E01-0263-20856 E01-0263-20856* E01-0263-20850 EOI-0263-20850* EOl-0263-20857
E01-0263-20857*
E01-0263-20851 E01-0263-20851* E01-0263-20858 E01-0263-20858*
Centre ID
0103216 0103216 0103192 0103192 0103220
0103220
0103196 0103196 0103224 0103224
Set Number
032601C 032601C 032601C 032601C 032601C
032601C
032601C 032601C 032601C 032601C
Extraction Analysis
Date Date
3/26/01
3/27/01
3/26/01
3/27/01
3/26/01
3/27/01
3/26/01
3/27/01
3/26/01
3/27/01
3/26/01
3/27/01
3/26/01
3/27/01
3/26/01
3/27/01
3/26/01
3/27/01
3/26/01
3/27/01
AVERAGE:
STANDARD DEVIATION:
Analyte Found (ppb)
248** 264** 26.5 28.6 78.9
79.0
287** 281** 50.1 49.2 52.1 23.1
* Duplicate Injection
LOQ = 10 ppb
** Possible contamination by yolk or outer shell component of sample and as per Sponsor Representative request were not used to calculate the average and standard deviation.
Centre A nalytical Laboratories, Inc.
Page 27 o f 102
Centre Study N o.: 023-063
Table XI. Summary of PFOS for Week 1 Mallard Albumen Control Samples
Sponsor ID
E01-0263-20852 E01-0263-20852* E01-0263-20853 E01-0263-20853* E01-0263-20854 E01-0263-20854* EOI-0263-20842 EOl-0263-20842* E01-0263-20843 E01-0263-20843*
Centre ID
0103201 0103201 0103205 0103205 0103209 0103209 0103161 0103161 0103165 0103165
Set Number
032601A 032601A 032601A 032601A 032601A 032601A 032601A 032601A 032601A 032601A
Extraction Analysis
Date Date
3/26/01
3/26-27/01
3/26/01
3/26-27/01
3/26/01
3/26-27/01
3/26/01
3/26-27/01
3/26/01
3/26-27/01
3/26/01
3/26-27/01
3/26/01
3/26-27/01
3/26/01
3/26-27/01
3/26/01
3/26-27/01
3/26/01
3/26-27/01
AVERAGE:
STANDARD DEVIATION:
Analyte Found (ppb)
NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ
Table XII. Summary of PFOS for Week 1 Mallard Albumen Samples at 6.2 ppm ai
Sponsor ID
E01-0263-20825 E01-0263-20825* E01-0263-20846 E01-0263-20846* E01-0263-20832
E01-0263-20832*
E01-0263-20847 E01-0263-20847* E01-0263-20848 E01-0263-20848*
Centre ID
0103093 0103093 0103177 0103177 0103121 0103121 0103181 0103181 0103185 0103185
Set Number
032601A 032601A 032601A 032601A 032601A
032601A
032601A 032601A 032601A 032601A
Extraction Analysis
Date Date
3/26/01 3/26/01 3/26/01
3/26-27/01 3/26-27/01 3/26-27/01
3/26/01 3/26/01
3/26/01
3/26/01 3/26/01 3/26/01
3/26-27/01 3/26-27/01
3/26-27/01
3/26-27/01 3/26-27/01 3/26-27/01
3/26/01
3/26-27/01
AVERAGE: STANDARD DEVIATION:
Analyte Found (ppb)
2.49 2.75 7.90 7.89 5.67
5.79
5.76 5.58 5.95 6.29 5.63 1.80
* Duplicate Injection
NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) LOQ = 10 ppb
Centre A nalytical Laboratories, Inc.
Page 28 o f 102
Centre Study N o.: 023-063
Table XIII. Summary of PFOS for Week 1 Mallard Albumen Samples at 17.6 ppm ai
Sponsor ID
E01-0263-20856 E01-0263-20856* E01-0263-20856 E01-0263-20856* E01-0263-20856 E01-0263-20856* E01-0263-20850 E01-0263-20850* E01-0263-20857 E01-0263-20857* E01-0263-20851 E01-0263-20851* E01-0263-20858 E01-0263-20858*
Centre ID
0103217 0103217 0103217 0103217 0103217 Dup 0103217 Dup 0103193 0103193 0103221 0103221 0103197 0103197 0103225 0103225
Set Number
032601D 032601D 033001C 033001C 033001C 033001C 032601A 032601A 032601A 032601A 032601A 032601A 032601A 032601A
Extraction Analysis
Date Date
3/26/01 3/26/01 3/30/01 3/30/01 3/30/01 3/30/01 3/26/01
3/27-28/01 3/27-28/01
4/2/01 4/2/01 4/2/01 4/2/01 3/26-27/01
3/26/01
3/26-27/01
3/26/01
3/26-27/01
3/26/01
3/26-27/01
3/26/01
3/26-27/01
3/26/01
3/26-27/01
3/26/01
3/26-27/01
3/26/01
3/26-27/01
AVERAGE:
STANDARD DEVIATION:
Analyte Found (ppb)
3550** 3490** 1388** 1369** 845** 771**
24.7 23.9 23.8 23.3 78.0** 69.7** 19.7 19.8 22.5 2.20
Table XIV. Summary of PFOS for Week 1 Mallard Yolk Control Samples
Sponsor ID
E01-0263-20852 EOI-0263-20852*
EOl-0263-20853
E01-0263-20853* E01-0263-20854 EOI-0263-20854* EOl-0263-20842 E01-0263-20842* E01-0263-20843 E01-0263-20843*
Centre ID
0103198 0103198
0103202
0103202 0103206 0103206 0103158 0103158 0103162 0103162
* Duplicate Injection
Set Number
032601B 032601B
0 3 2 6 0 IB
032601B 032601B 032601B 032601B 032601B 032601B 032601B
Extraction Date
Analysis Date
Analyte Found (ppb)
3/26/01
3/27/01
3/26/01
3/27/01
3/26/01
3/27/01
3/26/01
3/27/01
3/26/01
3/27/01
3/26/01
3/27/01
3/26/01
3/27/01
3/26/01
3/27/01
3/26/01
3/27/01
3/26/01
3/27/01
AVERAGE:
STANDARD DEVIATION:
NQ NQ
NQ
NQ NQ NQ NQ NQ NQ NQ NQ NQ
NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) LOQ = 10 ppb
** Possible contamination by yolk or outer shell component of sample and as per Sponsor
Representative request were not used to calculate the average and standard deviation.
Centre Analytical Laboratories, Inc.
Page 29 of 102
Centre Study N o.: 023-063
Table XV. Summary of PFOS for Week 1 Mallard Yolk Samples at 6.2 ppm ai
Sponsor ID
E01-0263-20825 E01-0263-20825* E01-0263-20846 EOI-0263-20846* EOI-0263-20832 EOI-0263-20832* EOI-0263-20847 E01-0263-20847* E01-0263-20848 E01-0263-20848*
Centre ID
0103090 0103090 0103174 0103174 0103118 0103118 0103178 0103178 0103182 0103182
Set Number
032601D 032601D 03260ID 03260ID 032601D 032601D 032601D 032601D 032601D 032601D
Extraction Analysis
Date Date
3/26/01
3/27-28/01
3/26/01
3/27-28/01
3/26/01
3/27-28/01
3/26/01
3/27-28/01
3/26/01
3/27-28/01
3/26/01
3/27-28/01
3/26/01
3/27-28/01
3/26/01
3/27-28/01
3/26/01
3/27-28/01
3/26/01
3/27-28/01
AVERAGE:
STANDARD DEVIATION:
Analyte Found (ppb)
2470 2580 12500 12100 4010 3890 10500 11100 12100 12100 8340 4450
Table XVI. Summary of PFOS for Week 1 Mallard Yolk Samples at 17.6 ppm ai
Sponsor ID
E01-0263-20856 EOI-0263-20856* EOl-0263-20850 EOI-0263-20850* EOl-0263-20857 E01-0263-20857*
EO1-0263-20851
E01-0263-20851* E01-0263-20858 E01-0263-20858*
Centre ID
0103214 0103214 0103190 0103190 0103218 0103218
0103194
0103194 0103222 0103222
* Duplicate Injection
LOQ = 10 ppb
Set Number
032601D 032601D 032601D 032601D 032601D 032601D
032601D
032601D 032601D 032601D
Extraction Analysis
Date Date
3/26/01
3/27-28/01
3/26/01
3/27-28/01
3/26/01
3/27-28/01
3/26/01
3/27-28/01
3/26/01
3/27-28/01
3/26/01
3/27-28/01
3/26/01
3/27-28/01
3/26/01
3/27-28/01
3/26/01
3/27-28/01
3/26/01
3/27-28/01
AVERAGE:
STANDARD DEVIATION:
Analyte Found (ppb)
30300 30000 40000 43000 57200 56600
51700
53100 61000 62000 48500 12000
Centre A nalytical Laboratories, Inc.
Page 30 o f 102
Centre Study N o.: 023-063
Table XVII. Summary of PFOS for Week 6 Mallard Membrane Control Samples
Sponsor ID
E01-0263-20993 E01-0263-20993* E01-0263-20994 E01-0263-20994* E01-0263-20995 E01-0263-20995* E01-0263-20975 EOI-0263-20975* EOI-0263-20997 EOl-0263-20997*
Centre ID
0103764 0103764 0103768 0103768 0103772 0103772 0103692 0103692 0103780 0103780
Set Number
032701C 032701C 032701C 032701C 032701C 032701C 032701C 032701C 032701C 032701C
Extraction Analysis
Analyte
Date Date
3/27/01
3/29/01
NQ
3/27/01
3/29/01
NQ
3/27/01
3/29/01
NQ
3/27/01
3/29/01
NQ
3/27/01
3/29/01
NQ
3/27/01
3/29/01
NQ
3/27/01
3/29/01
NQ
3/27/01
3/29/01
NQ
3/27/01
3/29/01
NQ
3/27/01
3/29/01 ____ NQ_____
AVERAGE:
NQ
STANDARD DEVIATION:
NQ
Table XVIII. Summary of PFOS for Week 6 Mallard Membrane Samples at 6.2 ppm ai
Sponsor ID
E01-0263-20979 E01-0263-20979* E01-0263-21001 E01-0263-21001* E01-0263-20988 E01-0263-20988*
E01-0263-20971
E01-0263-20971* EOI-0263-20989 E01-0263-20989*
Centre ID
0103708 0103708 0103796 0103796 0103744 0103744
0103676
0103676 0103748 0103748
* Duplicate Injection
Set Number
032701C 032701C 032701C 032701C 032701C 032701C
032701C
032701C 032701C 032701C
Extraction Analysis
Date Date
3/27/01 3/27/01 3/27/01 3/27/01 3/27/01 3/27/01
3/29/01 3/29/01 3/29/01 3/29/01 3/29/01 3/29/01
3/27/01
3/27/01 3/27/01 3/27/01
3/29/01
3/29/01 3/29/01 3/29/01
AVERAGE: STANDARD DEVIATION:
Analyte Found (ppb)
70.8 70.6 55.3 55.3 95.7 98.7
169**
171** 83.8 88.2 77.3 17.0
NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) LOQ = 10 ppb
** Possible contamination by yolk or outer shell component of sample and as per Sponsor Representative request were not used to calculate the average and standard deviation.
Centre A nalytical Laboratories, Inc.
Page 31 o f 102
Centre Study N o.: 023-063
Table XIX. Summary of PFOS for Week 6 Mallard Membrane Samples at 17.6 ppm ai
Sponsor ID
E01-0263-20991 E01-0263-20991* E01-0263-20990 E01-0263-20990* E0I-0263-21002 E01-0263-21002* E01-0263-21003 E01-0263-21003* E01-0263-21004 E01-0263-21004*
Centre ID
0103756 0103756 0103752 0103752 0103800 0103800 0103804 0103804 0103808 0103808
Set Number
032701CD 032701CD 032701CD 032701CD 032701CD 032701CD 032701CD 032701CD 032701CD 032701CD
Extraction Analysis
Date Date
3/27/01 3/27/01 3/27/01 3/27/01 3/27/01 3/27/01 3/27/01 3/27/01 3/27/01 3/27/01
3/29/01 3/29/01 3/29/01 3/29/01 3/29/01 3/29/01 3/29/01 3/29/01 3/29/01 3/29/01
AVERAGE: STANDARD DEVIATION:
Analyte Found (ppb)
434 431 365 370 347 339 306 307 259 261 342 61.2
Table XX. Summary of PFOS for Week 6 Mallard Albumen Control Samples
Sponsor ID
E01-0263-20993 E01-0263-20993* E01-0263-20994 E01-0263-20994* E01-0263-20995 E01-0263-20995*
E01-0263-20975
E01-0263-20975* E01-0263-20997 E01-0263-20997*
Centre ID
0103765 0103765 0103769 0103769 0103773 0103773
0103693
0103693 0103781 0103781
* Duplicate Injection
Set Number
032701B 032701B 032701B 032701B 032701B 032701B
032701B
032701B 032701B 032701B
Extraction Analysis
Date Date
3/27/01 3/27/01 3/27/01 3/27/01 3/27/01
3/28-29/01 3/28-29/01 3/28-29/01 3/28-29/01 3/28-29/01
3/27/01
3/27/01
3/28-29/01
3/28-29/01
3/27/01 3/27/01 3/27/01
3/28-29/01 3/28-29/01 3/28-29/01
AVERAGE:
STANDARD DEVIATION:
Analyte Found (ppb)
NQ NQ NQ NQ NQ NQ
NQ
NQ NQ NQ NQ NQ
NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) LOQ = 10 ppb
Centre A nalytical Laboratories, Inc.
Page 32 o f 102
Centre Study N o.: 023-063
Table XXI. Summary of PFOS for Week 6 Mallard Albumen Samples at 6.2 ppm ai
Sponsor ID
E01-0263-20979 E01-0263-20979* E01-0263-21001 E01-0263-21001* E01-0263-20988 E01-0263-20988* E01-0263-20971 E01-0263-20971* E01-0263-20989 E01-0263-20989*
Centre ID
0103709 0103709 0103797 0103797 0103745 0103745 0103677 0103677 0103749 0103749
Set Number
032701B 032701B 032701B 032701B 032701B 032701B 032701B 032701B 032701B 032701B
Extraction Analysis
Date Date
3/27/01 3/28-29/01
3/27/01 3/28-29/01
3/27/01 3/28-29/01
3/27/01 3/28-29/01
3/27/01 3/28-29/01
3/27/01
3/28-29/01
3/27/01
3/28-29/01
3/27/01
3/28-29/01
3/27/01 3/28-29/01
3/27/01 3/28-29/01
AVERAGE:
STANDARD DEVIATION:
Analyte Found (ppb)
5.83 6.23 7.92 8.04 6.56 6.26 7.02 6.81 8.72 8.33 7.19 0.987
Table XXII. Summary of PFOS for Week 6 Mallard Albumen Samples at 17.6 ppm ai
Sponsor ID
E01-0263-20991 E01-0263-20991* E01-0263-20990 E01-0263-20990* E01-0263-21002 E01-0263-21002*
E01-0263-21003
E01-0263-21003* E01-0263-21004 E01-0263-21004*
Centre ID
0103757 0103757 0103753 0103753 0103801 0103801
0103805
0103805 0103809 0103809
* Duplicate Injection
LOQ = 10 ppb
Set Number
03270IB 032701B 032701B 032701B 032701B 032701B
032701B
032701B 032701B 032701B
Extraction Analysis
Date Date
3/27/01 3/27/01 3/27/01 3/27/01 3/27/01 3/27/01
3/27/01
3/27/01 3/27/01 3/27/01
3/28-29/01 3/28-29/01 3/28-29/01 3/28-29/01 3/28-29/01 3/28-29/01
3/28-29/01
3/28-29/01 3/28-29/01 3/28-29/01
AVERAGE: STANDARD DEVIATION:
Analyte Found(ppb)
25.3 24.9 14.1 14.7 19.7 19.4
23.6
24.0 19.1 17.2 20.2 4.12
Centre A nalytical Laboratories, Inc.
Page 33 o f 102
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Table XXIII. Summary of PFOS for Week 6 Mallard Yolk Control Samples
Sponsor ID
E01-0263-20993 E01-0263-20993* E01-0263-20994 E01-0263-20994* E01-0263-20995 E01-0263-20995* E01-0263-20975 E01-0263-20975* E01-0263-20997 E01-0263-20997*
Centre ID
0103762 0103762 0103766 0103766 0103770 0103770 0103690 0103690 0103778 0103778
Set Number
032701A 032701A 032701A 032701A 032701A 032701A 032701A 032701A 032701A 032701A
Extraction Analysis
Analyte
Date Date Found (ppb)
3/27/01
3/28/01
NQ
3/27/01 3/27/01
3/28/01 3/28/01
NQ NQ
3/27/01 3/27/01 3/27/01 3/27/01
3/28/01 3/28/01 3/28/01 3/28/01
NQ NQ NQ NQ
3/27/01
3/28/01
NQ
3/27/01 3/27/01
3/28/01
NQ
3/28/01 _____ NQ____ AVERAGE: NQ
STANDARD DEVIATION: NQ
Table XXIV. Summary of PFOS for Week 6 Mallard Yolk Samples at 6.2 ppm ai
Sponsor ID
E01-0263-20979 E01-0263-20979* E01-0263-21001 E01-0263-21001* E01-0263-20988 E01-0263-20988*
EO1-0263-20971
E01-0263-20971* E01-0263-20989 E01-0263-20989*
Centre ID
0103706 0103706 0103794 0103794 0103742 0103742
0103674
0103674 0103746 0103746
* Duplicate Injection
Set Number
032701AD 032701AD 032701AD 032701AD 032701AD 032701AD
032701AD
032701AD 032701AD 032701AD
Extraction Analysis
Date Date
3/27/01
3/29/01
3/27/01
3/29/01
3/27/01
3/29/01
3/27/01
3/29/01
3/27/01
3/29/01
3/27/01
3/29/01
3/27/01
3/29/01
3/27/01
3/29/01
3/27/01
3/29/01
3/27/01
3/29/01
AVERAGE:
STANDARD DEVIATION:
Analyte Found(ppb)
16100 15700 9860 10400 11200 10600
16800
16900 27200 26600 16100 6330
NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) LOQ = 10 ppb
Centre A nalytical Laboratories, Inc.
Page 34 o f 102
Centre Study N o.: 023-063
Table XXV. Summary of PFOS for Week 6 Mallard Yolk Samples at 17.6 ppm ai
Sponsor ID
E01-0263-20991 E01-0263-20991* E01-0263-20990 E01-0263-20990* E01-0263-21002 E01-0263-21002* E01-0263-21003 E01-0263-21003* E01-0263-21004 E01-0263-21004*
Centre ID
0103754 0103754 0103750 0103750 0103798 0103798 0103802 0103802 0103806 0103806
Set Number
032701AD 032701AD 032701AD 032701AD 032701AD 032701AD 032701AD 032701AD 032701AD 032701AD
Extraction Analysis
Date Date
3/27/01
3/29/01
3/27/01
3/29/01
3/27/01
3/29/01
3/27/01
3/29/01
3/27/01
3/29/01
3/27/01
3/29/01
3/27/01
3/29/01
3/27/01
3/29/01
3/27/01
3/29/01
3/27/01
3/29/01
AVERAGE:
STANDARD DEVIATION:
Analyte Found (ppb)
41900 40700 63000 63500 57500 57300 43100 43600 46600 49800 50700 8870
Table XXVI. Summary of PFOS in Quail Membrane Blanks
Sponsor ID
na na na na
Centre ID
0003798 Blank A 0003798 Blank A* 0003798 Blank A 0003798 Blank A*
Set Extraction Analysis
Number
Date
Date
032901A
3/29/01
3/31/01
032901A
3/29/01
3/31/01
033001BR 3/30/01
4/2/01
033001BR 3/30/01
4/2/01
AVERAGE:
STANDARD DEVIATION:
Analyte F'ound (ppb)
NQ NQ NQ NQ NQ NQ
Table XXVII. Summary of PFOS in Quail Albumen Blanks
Sponsor ID
na na na na
Centre ID
0003796 Blank A 0003796 Blank A* 0003796 Blank A 0003796 Blank A*
* Duplicate Injection
Set Number
032801B 032801B 033001A 033001A
Extraction Analysis
Date Date
3/28/01
3/30/01
3/28/01
3/30/01
3/30/01
4/1/01
3/30/01
4/1/01
AVERAGE:
STANDARD DEVIATION:
Analyte Found (ppb)
NQ NQ NQ NQ NQ NQ
NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) LOQ = 10 ppb
Centre A nalytical Laboratories, Inc.
Page 35 o f 102
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Table XXVIII. Summary of PFOS in Quail Yolk Blanks
Sponsor ID
na na na na
Centre ID
0003795 Blank A 0003795 Blank A* 0003795 Blank A 0003795 Blank A*
Set Number
032801A 032801A 032901B 032901B
Extraction Analysis
Date Date
3/28/01
3/30/01
3/28/01 3/29/01 3/29/01
3/30/01 3/31-4/1/01 3/31-4/1/01
AVERAGE:
STANDARD DEVIATION:
Analyte Found (ppb)
NQ NQ ND ND NQ NQ
Table XXIX. Summary of PFOS Recoveries in Quail Membrane
Sponsor ID
na na na na na na na na
Centre ID
0003798 Spk A 0003798 Spk A* 0003798 SpkB 0003798 SpkB* 0003798 Spk A 0003798 Spk A* 0003798 SpkB 0003798 SpkB*
Set Number
032901A 032901A 032901A 032901A 033001BR 033001BR 033001B 033001B
Extraction Analysis
Amt
Date Date Added (ppb)
3/29/01 3/29/01
3/31/01 3/31/01
10 10
3/29/01 3/31/01
986
3/29/01 3/31/01
986
3/30/01
4/2/01
10
3/30/01
4/2/01
10
3/30/01
4/2/01
98
3/30/01
4/2/01
98
AVERAGE:
STANDARD DEVIATION: RELATIVE STANDARD DEVIATION:
% Recovery
102 104 88 85 114 116 108 101 102 11 11
* Duplicate Injection
NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) ND = No peak detected.
LOQ = 10 ppb
Centre A nalytical Laboratories, Inc.
Page 36 o f 102
Centre Study N o.: 023-063
Table XXX. Summary of PFOS Recoveries in Quail Albumen
Sponsor ID
na na na na na na na na
Centre ID
0003796 Spk A 0003796 Spk A* 0003796 SpkB 0003796 SpkB* 0003796 Spk A 0003796 Spk A* 0003796 SpkB 0003796 SpkB*
Set Number
03280IB 032801B 032801B 032801B 033001A 033001A 033001A 033001A
Extraction Analysis
Amt
%
Date _ ^ D a te _ _ Added (ppb) ^Recovery
3/28/01
3/30/01
10
82
3/28/01
3/30/01
10
79
3/28/01
3/30/01
9963
80
3/28/01
3/30/01
9963
83
3/30/01
4/1/01
10
82
3/30/01 3/30/01 3/30/01
4/1/01 4/1/01 4/1/01
10 99 99
88 93 91
AVERAGE: 85
STANDARD DEVIATION: RELATIVE STANDARD DEVIATION:
5 6
Table XXXI. Summary of PFOS Recoveries in Quail Yolk
Sponsor ID
na na na na na na na na
Centre ID
0003795 Spk A 0003795 Spk A* 0003795 SpkB 0003795 SpkB* 0003795 Spk A 0003795 Spk A* 0003795 SpkB 0003795 SpkB*
Set Number
032801A 032801A 032801A 032801A 032901B 032901B 032901D 032901D
Extraction Analysis
Amt
Date Date
3/28/01 3/30/01
10
3/28/01 3/30/01
10
3/28/01 3/30/01
9623
3/28/01 3/30/01
9623
3/29/01 3/31-4/1/01
10
3/29/01 3/31-4/1/01
10
3/29/01 4/1-2/01
98834
3/29/01 4/1-2/01
98834
AVERAGE
STANDARD DEVIATION
RELATIVE STANDARD DEVIATION
* Duplicate Injection
LOQ = 10 ppb
%
102 104 98 101 87 87 94 93 96 7 7
Centre A nalytical Laboratories, Inc.
Page 37 o f 102
Centre Study N o.: 023-063
Table XXXII. Summary of PFOS for Week 1 Quail Membrane Control
Samples
Sponsor ID
Centre ID
Set Number
Extraction Date
Analysis Date
Analyte Found (ppb)
E01-0259-20506 E01-0259-20506* E01-0259-20507 E01-0259-20507* E01-0259-20520 E01-0259-20520* E01-0259-20521 E01-0259-20521* E01-0259-20509 E01-0259-20509*
0101987 0101987 0101991 0101991 0102043 0102043 0102047 0102047 0101999 0101999
032901A 032901A 032901A 032901A 032901A 032901A 032901A 032901A 032901A 032901A
3/29/01
3/31/01
3/29/01
3/31/01
3/29/01
3/31/01
3/29/01
3/31/01
3/29/01
3/31/01
3/29/01
3/31/01
3/29/01
3/31/01
3/29/01
3/31/01
3/29/01
3/31/01
3/29/01
3/31/01
AVERAGE:
STANDARD DEVIATION:
5.20** 5.57**
NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ
Table XXXIII. Summary of PFOS for Week 1 Quail Membrane
Samples at 6.2 ppm ai
Sponsor
Centre
Set
Extraction Analysis
Analyte
ID
ID
Number
Date
Date Found (ppb)
E01-0259-20526 E01-0259-20526* E01-0259-20514 E01-0259-20514* E01-0259-20527
0102067 0102067 0102019 0102019 0102071
032901D 032901D 032901A 032901A 032901A
3/29/01 3/29/01 3/29/01 3/29/01 3/29/01
4/1-2/01 4/1-2/01 3/31/01 3/31/01 3/31/01
18800** 17900**
127 129 123
E01-0259-20527* E01-0259-20515 E01-0259-20515* E01-0259-20528 E01-0259-20528*
0102071 0102023 0102023 0102075 0102075
032901A 032901A 032901A 032901D 032901D
3/29/01 3/29/01 3/29/01 3/29/01 3/29/01
3/31/01 3/31/01 3/31/01 4/1-2/01 4/1-2/01
116 172 172 11500** 11500**
AVERAGE: 140
* Duplicate Injection
STANDARD DEVIATION: 25.3
NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) LOQ = 10 ppb
** Possible contamination by yolk or outer shell component of sample since outer shell was not washed prior to separation of internal components. As per Sponsor Representative request these values were not used to calculate the average and standard deviation.
For residues recorded as NQ, half of the concentration of the lowest standard (0.5 ppb) was used to calculate the average and standard deviation.
Centre A nalytical Laboratories, Inc.
Page 38 o f 102
Centre Study N o.: 023-063
Table XXXIV. Summary of PFOS for Week 1 Quail Membrane Samples at 17.6 ppm ai
Sponsor ID
E01-0259-20529 E01-0259-20529* E01-0259-20531 E01-0259-20531* E01-0259-20532 E01-0259-20532* E01-0259-20518 E01-0259-20518* E01-0259-20533 E01-0259-20533*
Centre ID
0102079 0102079 0102087 0102087 0102091 0102091 0102035 0102035 0102095 0102095
Set Number
032901D 032901D 032901D 032901D 032901D 032901D 032901D 032901D 032901D 03290ID
Extraction Analysis
Date Date
3/29/01
4/1-2/01
3/29/01
4/1-2/01
3/29/01
4/1-2/01
3/29/01
4/1-2/01
3/29/01
4/1-2/01
3/29/01
4/1-2/01
3/29/01
4/1-2/01
3/29/01
4/1-2/01
3/29/01
4/1-2/01
3/29/01
4/1-2/01
AVERAGE:
STANDARD DEVIATION:
Analyte Found (ppb)
20900 20400 11000 11100 14300 13900 12500 12600 6880 6990 13100 4730
Table XXXV. Summary of PFOS for Week 1 Quail Albumen Control Samples
Sponsor ID
E01-0259-20506 E01-0259-20506* E01-0259-20507 E01-0259-20507* E01-0259-20520 EOI-0259-20520*
EOI-0259-20521
EOl-0259-20521* EOl-0259-20509 E01-0259-20509*
Centre ID
0101988 0101988 0101992 0101992 0102044 0102044
0102048
0102048 0102000 0102000
Set Number
032801B 032801B 03280IB 032801B 032801B 032801B
03280IB
032801B 03280IB 032801B
Extraction Analysis
Date Date
3/28/01
3/30/01
3/28/01
3/30/01
3/28/01
3/30/01
3/28/01
3/30/01
3/28/01
3/30/01
3/28/01
3/30/01
3/28/01
3/30/01
3/28/01
3/30/01
3/28/01
3/30/01
3/28/01
3/30/01
AVERAGE:
STANDARD DEVIATION:
Analyte Found (ppb)
NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ
* Duplicate Injection
NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/noL)) LOQ = 10 ppb
Centre A nalytical Laboratories, Inc.
Page 39 o f 102
Centre Study N o.: 023-063
Table XXXVI. Summary of PFOS for Week 1 Quail Albumen Samples at 6.2 ppm ai
Sponsor ID
E01-0259-20526 E01-0259-20526* E01-0259-20514 E01-0259-20514* EOI-0259-20527 EOl-0259-20527* E01-0259-20515 E01-0259-20515* E01-0259-20528 E01-0259-20528*
Centre ID
0102068 0102068 0102020 0102020 0102072 0102072 0102024 0102024 0102076 0102076
Set Number
032801B 032801B 032801B 032801B 032801B 032801B 032801B 032801B 032801B 032801B
Extraction Date
Analysis Date
Analyte !Found (ppb)
3/28/01
3/30-31/01
3/28/01
3/30-31/01
3/28/01
3/30-31/01
3/28/01
3/30-31/01
3/28/01
3/30-31/01
3/28/01
3/30-31/01
3/28/01
3/30-31/01
3/28/01
3/30-31/01
3/28/01
3/30-31/01
3/28/01
3/30-31/01
AVERAGE:
STANDARD DEVIATION:
6.03 5.43 2.43 2.78 2.94 3.48 5.87 6.08 93.9** 93.7** 4.38 1.61
Table XXXVII. Summary of PFOS for Week 1 Quail Albumen Samples at 17.6 ppm ai
Sponsor ID
E01-0259-20529 E01-0259-20529* E01-0259-20531 E01-0259-20531* E01-0259-20532 E01-0259-20532* EO1-0259-20518 E01-0259-20518* E01-0259-20533 E01-0259-20533*
Centre ID
0102080 0102080 0102088 0102088 0102092 0102092 0102036 0102036 0102096 0102096
* Duplicate Injection
LOQ = 10 ppb
Set Number
03280IB 032801B 032801B 03280IB 032801B 032801B 032801B 032801B 032801B 032801B
Extraction Analysis
Date Date
3/28/01 3/28/01 3/28/01 3/28/01 3/28/01 3/28/01 3/28/01 3/28/01 3/28/01 3/28/01
3/30-31/01 3/30-31/01 3/30-31/01 3/30-31/01 3/30-31/01 3/30-31/01 3/30-31/01 3/30-31/01 3/30-31/01 3/30-31/01 AVERAGE:
STANDARD DEVIATION:
Analyte
17.9 17.3 9.97 10.5 10.7 11.0 32.3 31.6 9.07 10.1 16.0 8.92
Centre A nalytical Laboratories, Inc.
Page 40 o f 102
Centre Study N o.: 023-063
Table XXXVIII. Summary of PFOS for Week 1 Quail Yolk Control Samples
Sponsor ID
E01-0259-20506 E01-0259-20506* E01-0259-20507 E01-0259-20507* E01-0259-20520 E01-0259-20520* E01-0259-20521 E01-0259-20521* E01-0259-20509 E01-0259-20509*
Centre ID
0101985 0101985 0101989 0101989 0102041 0102041 0102045 0102045 0101997 0101997
Set
Number
032801A 032801A 032801A 032801A 032801A 032801A 032801A 032801A 032801A 032801A
Extraction Analysis
Analyte
Date Date _^otmd(npb)
3/28/01
3/30/01
NQ
3/28/01 3/28/01 3/28/01 3/28/01 3/28/01
3/30/01 3/30/01 3/30/01 3/30/01 3/30/01
NQ NQ NQ NQ NQ
3/28/01 3/28/01 3/28/01 3/28/01
3/30/01 3/30/01 3/30/01 3/30/01
NQ NQ NQ NQ
AVERAGE: NQ
STANDARD DEVIATION: NQ
Table XXXIX. Summary of PFOS for Week 1 Quail Yolk Samples at 6.2 ppmai
Sponsor ID
E01-0259-20526 E01-0259-20526* E01-0259-20514 E01-0259-20514* E01-0259-20527 E01-0259-20527* E01-0259-20515 E01-0259-20515* E01-0259-20528 E01-0259-20528*
Centre ID
0102065 0102065 0102017 0102017 0102069 0102069 0102021 0102021 0102073 0102073
* Duplicate Injection
Set Number
032801AD 032801AD 032801AD 032801AD 032801AD 032801AD 032801AD 032801AD 032801AD 032801AD
Extraction Analysis
Date Date
3/28/01 3/28/01 3/28/01 3/28/01 3/28/01 3/28/01 3/28/01 3/28/01 3/28/01 3/28/01
3/31/01 3/31/01 3/31/01 3/31/01 3/31/01 3/31/01 3/31/01 3/31/01 3/31/01 3/31/01
AVERAGE: STANDARD DEVIATION:
Analyte Found (ppb)
17500 17500 8530 8410 12100 12200 13800 13800 11700 10900 12600 3150
NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) LOQ = 10 ppb
Centre A nalytical Laboratories, Inc.
Page 41 o f 102
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Table XL. Summary of PFOS for Week 1 Quail Yolk Samples at 17.6
ppmai
Sponsor ID
Centre ID
Set Number
Extraction Date
Analysis Date
Analyte Found (ppb)
E01-0259-20529 E01-0259-20529* E01-0259-2053I E01-0259-20531* E01-0259-20532 E01-0259-20532* E01-0259-20518 E01-0259-20518* E01-0259-20533 E01-0259-20533*
0102077 0102077 0102085 0102085 0102089 0102089 0102033 0102033 0102093 0102093
032801AD 032801AD 032801AD 032801AD 032801AD 032801AD 032801AD 032801AD 032801AD 032801AD
3/28/01
3/31/01
3/28/01
3/31/01
3/28/01
3/31/01
3/28/01
3/31/01
3/28/01
3/31/01
3/28/01
3/31/01
3/28/01
3/31/01
3/28/01
3/31/01
3/28/01
3/31/01
3/28/01
3/31/01
AVERAGE:
STANDARD DEVIATION:
42500 41200 33200 31100 37400 38900 38100 38300 43000 43600 38700 4130
Table XLI. Summary of PFOS for Week 6 Quail Membrane Control
Samples
Sponsor
Centre
Set
Extraction Analysis
Analyte
ID ID Number Date Date Found (pp
E01-0259-20735 E01-0259-20735* E01-0259-20736 E01-0259-20736* E01-0259-20737 E01-0259-20737*
0102903 0102903 0102907 0102907 0102911 0102911
033001B 033001B 033001B 033001B 033001B 033001B
3/30/01 3/30/01 3/30/01 3/30/01 3/30/01 3/30/01
4/2/01 4/2/01 4/2/01 4/2/01 4/2/01 4/2/01
NQ NQ 9.85** 8.93** 15.6** 14.4**
E01-0259-20720 E01-0259-20720* E01-0259-20738 E01-0259-20738*
0102843 0102843 0102915 0102915
* Duplicate Injection
033001B 033001B 033001B 033001B
3/30/01
4/2/01
3/30/01
4/2/01
3/30/01
4/2/01
3/30/01
4/2/01
AVERAGE:
STANDARD DEVIATION:
21.8** 24.4** 6.51** 6.30** NQ NQ
NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) LOQ = 10 ppb
** Possible contamination by yolk or outer shell component of sample since outer shell was not washed prior to separation of internal components. As per Sponsor Representative request these values were not used to calculate the average and standard deviation.
Centre A nalytical Laboratories, Inc.
Page 42 o f 102
Centre Study N o.: 023-063
Table XLII. Summary of PFOS for Week 6 Quail Membrane Samples at 6.2 ppm ai
ID
E01-0259-20742 E01-0259-20742* EOI-0259-20743 E01-0259-20743* E01-0259-20744 EOI-0259-20744* EOl-0259-20745 E01-0259-20745* E01-0259-20746 E01-0259-20746*
ID
0102931 0102931 0102935 0102935 0102939 0102939 0102943 0102943 0102947 0102947
Number
033001B 033001B 033001B 033001B 033001B 033001B 033001B 033001B 03300IB 033001B
Date Date
3/30/01
4/2/01
3/30/01
4/2/01
3/30/01
4/2/01
3/30/01
4/2/01
3/30/01
4/2/01
3/30/01
4/2/01
3/30/01
4/2/01
3/30/01
4/2/01
3/30/01
4/2/01
3/30/01
4/2/01
AVERAGE:
STANDARD DEVIATION:
Found (ppb)
120 119 213 208 123 116 109 116 134 134 139 38.4
Table XLIII. Summary of PFOS for Week 6 Quail Membrane Samples at 17.6 ppm ai
Sponsor ID
E01-0259-20747 E01-0259-20747* E01-0259-20748 E01-0259-20748* E01-0259-20749 E01-0259-20749* E01-0259-20750
E01-0259-20750*
E01-0259-20751 E01-0259-20751*
Centre ID
0102951 0102951 0102955 0102955 0102959 0102959 0102963 0102963 0102967 0102967
* Duplicate Injection
LOQ = 10 ppb
Set Number
033001BD 033001BD 033001BD 033001BD 033001BD 033001BD 033001B 033001B 033001B 033001B
Extraction Date
Analysis Date
3/30/01
4/2/01
3/30/01
4/2/01
3/30/01
4/2/01
3/30/01
4/2/01
3/30/01
4/2/01
3/30/01
4/2/01
3/30/01
4/2/01
3/30/01
4/2/01
3/30/01
4/2/01
3/30/01
4/2/01
AVERAGE:
STANDARD DEVIATION:
Analyte
Found (ppb)
405 389 318 318 394 400 262 250 186 185 311 86.6
Centre A nalytical Laboratories, Inc.
Page 43 o f 102
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Table XLIV. Summary of PFOS for Week 6 Quail Albumen Control Samples
Sponsor ID
E01-0259-20735 E01-0259-20735* E01-0259-20736 E01-0259-20736* E01-0259-20737 E01-0259-20737* E01-0259-20720 E01-0259-20720* E01-0259-20738 E01-0259-20738*
Centre ID
0102904 0102904 0102908 0102908 0102912 0102912 0102844 0102844 0102916 0102916
Set Number
033001A 033001A 033001A 033001A 033001A 033001A 033001A 033001A 033001A 033001A
Extraction Analysis
Date Date
3/30/01 3/30/01 3/30/01 3/30/01 3/30/01 3/30/01 3/30/01 3/30/01 3/30/01 3/30/01
4/1/01 4/1/01 4/1/01 4/1/01 4/1/01 4/1/01 4/1/01 4/1/01 4/1/01 4/1/01
AVERAGE: STANDARD DEVIATION:
Analyte Found (ppb)
NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ
Table XLV. Summary of PFOS for Week 6 Quail Albumen Samples at 6.2 ppm ai
Sponsor ID
E01-0259-20742 E01-0259-20742* E01-0259-20743 E01-0259-20743* E01-0259-20744 E01-0259-20744*
E01-0259-20745
E01-0259-20745* E01-0259-20746 E01-0259-20746*
Centre ID
0102932 0102932 0102936 0102936 0102940 0102940
0102944
0102944 0102948 0102948
* Duplicate Injection
Set Number
033001A 033001A 033001A 033001A 033001A 033001A
033001A
033001A 033001A 033001A
Extraction Analysis
Date Date
3/30/01 3/30/01 3/30/01 3/30/01 3/30/01
4/1/01 4/1/01 4/1/01 4/1/01 4/1/01
3/30/01
3/30/01
3/30/01 3/30/01 3/30/01
4/1/01
4/1/01
4/1/01 4/1/01 4/1/01
AVERAGE:
STANDARD DEVIATION:
Analyte Found(ppb)
5.34 5.93 6.19 6.91 6.82 6.82
5.26
5.15 3.56 4.34 5.63 1.12
NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) LOQ = 10 ppb
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Page 44 o f 102
Centre Study No.: 023-063
Table XLVI. Summary of PFOS for Week 6 Quail Albumen Samples at
17.6 ppm ai
Sponsor ID
Centre ID
Set Number
Extraction Date
Analysis Date
Analyte Found (ppb)
E01-0259-20747 E01-0259-20747* E01-0259-20748 E01-0259-20748* E01-0259-20749 E01-0259-20749* E01-0259-20750 E01-0259-20750* E01-0259-20751 EOI-0259-20751*
0102952 0102952 0102956 0102956 0102960 0102960 0102964 0102964 0102968 0102968
033001A 033001A 033001A 033001A 033001A 033001A 033001A 033001A 033001A 033001A
3/30/01
4/1/01
3/30/01
4/1/01
3/30/01
4/1/01
3/30/01
4/1/01
3/30/01
4/1/01
3/30/01
4/1/01
3/30/01
4/1/01
3/30/01
4/1/01
3/30/01
4/1/01
3/30/01
4/1/01
AVERAGE:
STANDARD DEVIATION:
9.81 9.84 13.2 13.1 14.6 14.2 16.3 16.7 15.6 15.4 13.9 2.44
Table XLVII. Summary of PFOS for Week 6 Quail Yolk Control
Samples
Sponsor ID
Centre ID
Set Number
Extraction Date
Analysis Date
Analyte Found (ppb)
E01-0259-20735 E01-0259-20735* E01-0259-20736 E01-0259-20736* E01-0259-20737 E01-0259-20737* E01-0259-20720 E01-0259-20720* E01-0259-20738 E01-0259-20738*
0102901 0102901 0102905 0102905 0102909 0102909 0102841 0102841 0102913 0102913
* Duplicate Injection
032901B 032901B 032901B 032901B 032901B 032901B 032901B 032901B 032901B 032901B
3/29/01 3/31-4/1/01 3/29/01 3/31-4/1/01 3/29/01 3/31-4/1/01 3/29/01 3/31-4/1/01 3/29/01 3/31-4/1/01 3/29/01 3/31-4/1/01 3/29/01 3/31-4/1/01 3/29/01 3/31-4/1/01 3/29/01 3/31-4/1/01 3/29/01 3/31-4/1/01
AVERAGE: STANDARD DEVIATION:
NQ NQ NQ NQ 51.5** 51.1** NQ NQ ND ND NQ NQ
NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) ND = No peak detected LOQ = 10 ppb
** Possible contamination by outer shell component of sample since outer shell was not washed prior to separation of internal components. As per Sponsor Representative request these values were not used to calculate the average and standard deviation.
Centre A nalytical Laboratories, Inc.
Page 45 o f 102
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Table XLVIII. Summary of PFOS for Week 6 Quail Yolk Samples at 6.2 ppm ai
Sponsor ID
E01-0259-20742 EOI-0259-20742* EOl-0259-20743 E01-0259-20743* E01-0259-20744 E01-0259-20744* E01-0259-20745 E01-0259-20745* E01-0259-20746 E01-0259-20746*
Centre ID
0102929 0102929 0102933 0102933 0102937 0102937 0102941 0102941 0102945 0102945
Set Number
032901D 032901D 032901D 032901D 032901D 032901D 032901D 032901D 032901D 032901D
Extraction Analysis
Date Date
3/29/01
4/1-2/01
3/29/01
4/1-2/01
3/29/01
4/1-2/01
3/29/01
4/1-2/01
3/29/01
4/1-2/01
3/29/01
4/1-2/01
3/29/01
4/1-2/01
3/29/01
4/1-2/01
3/29/01
4/1-2/01
3/29/01
4/1-2/01
AVERAGE:
STANDARD DEVIATION:
Analyte Found (ppb)
38700\ 40500 37100^ 361(Xr 236002360o r 400004060CT 2460025800^ 33100 7610
Table IL. Summary of PFOS for Week 6 Quail Yolk Samples at 17.6 ppmai
Sponsor
ID
E01-0259-20747 E01-0259-20747* E01-0259-20748 E01-0259-20748* E01-0259-20749 E01-0259-20749* EO1-0259-20750 E01-0259-20750* E01-0259-20751 E01-0259-20751*
Centre ID
0102949 0102949 0102953 0102953 0102957 0102957 0102961 0102961 0102965 0102965
* Duplicate Injection
Set Number
032901D 032901D 032901D 032901D 032901D 032901D 032901D 032901D 032901D 032901D
Extraction Analysis
Date Date
3/29/01
4/1-2/01
3/29/01
4/1-2/01
3/29/01
4/1-2/01
3/29/01
4/1-2/01
3/29/01
4/1-2/01
3/29/01
4/1-2/01
3/29/01
4/1-2/01
3/29/01
4/1-2/01
3/29/01
4/1-2/01
3/29/01
4/1-2/01
AVERAGE:
STANDARD DEVIATION:
Analyte Found (ppb)
53900 v 54400 65500 64800" 46400 465O0S 59400n 59300^ 56800 57800 56500 6510
NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL))
Centre A nalytical Laboratories, Inc.
Page 46 o f 102
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Table L. Summary of Moisture Determinations for Mallard Membrane Samples
Sponsor ID
E01-0263-20852 E01-0263-20853 E01-0263-20854 E01-0263-20842 E01-0263-20843 E01-0263-20825 E01-0263-20846 E01-0263-20832 E01-0263-20847 E01-0263-20848 E01-0263-20856 E01-0263-20850 E01-0263-20857 E01-0263-20851 E01-0263-20858 E01-0263-20993 E01-0263-20994 E01-0263-20995 E01-0263-20975 E01-0263-20997 E01-0263-20979 E01-0263-21001 E01-0263-20988 E01-0263-20971 E01-0263-20989 E01-0263-20991 E01-0263-20990 E01-0263-21002 E01-0263-21003 E01-0263-21004
Centre
ID
0103200 0103204 0103208 0103160 0103164 0103092 0103176 0103120 0103180 0103184 0103216 0103192 0103220 0103196 0103224 0103764 0103768 0103772 0103692 0103780 0103708 0103796 0103744 0103676 0103748
0103756
0103752 0103800 0103804 0103808
Average % Moisture
80.0 55.0 75.0 80.0 80.0 70.0 70.0 75.0 65.0 60.0 75.0 50.0 85.0 85.0 85.0 75.0 71.4 80.0 65.0 80.0 75.0 65.0 75.0 90.0 80.0
75.0
70.0 70.0 75.0 65.0
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Table LI. Summary of Moisture Determinations for Mallard Albumen Samples
Sponsor ID
E01-0263-20852 E01-0263-20853 E01-0263-20854 E01-0263-20842 E01-0263-20843 E01-0263-20825 E01-0263-20846 E01-0263-20832 E01-0263-20847 E01-0263-20848 E01-0263-20856 E01-0263-20850 E01-0263-20857 E01-0263-20851 E01-0263-20858 E01-0263-20993 E01-0263-20994 E01-0263-20995 E01-0263-20975 E01-0263-20997 E01-0263-20979 E01-0263-21001 E01-0263-20988 E01-0263-20971 E01-0263-20989
E01-0263-20991
E01-0263-20990 E01-0263-21002 E01-0263-21003 E01-0263-21004
Centre ID
0103201 0103205 0103209 0103161 0103165 0103093 0103177 0103121 0103181 0103185 0103217 0103193 0103221 0103197 0103225 0103765 0103769 0103773 0103693 0103781 0103709 0103797 0103745 0103677 0103749
0103757
0103753 0103801 0103805 0103809
Average % Moisture
90.0 88.0 87.0 85.0 87.0 82.0 84.0 86.0 89.0 88.0 85.0 85.0 84.0 83.0 85.0 88.0 85.0 85.0 84.0 90.0 88.0 84.0 87.0 83.0 87.0
84.0
86.0 83.0 82.0 87.0
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Table LII. Summary of Moisture Determinations for Mallard Yolk Samples
Sponsor ID
E01-0263-20852 E01-0263-20853 E01-0263-20854 E01-0263-20842 E01-0263-20843 E01-0263-20825 E01-0263-20846 E01-0263-20832 E01-0263-20847 E01-0263-20848 E01-0263-20856 EO1-0263-20850 EO1-0263-20857 EO1-0263-20851 EO1-0263-20858 E01-0263-20993 E01-0263-20994 E01-0263-20995 EO1-0263-20975 EO1-0263-20997 EO1-0263-20979 E01-0263-21001 E01-0263-20988 EO1-0263-20971 EO1-0263-20989
E01-0263-20991
E01-0263-20990 EO1-0263-21002 E01-0263-21003 E01-0263-21004
Centre ID
0103198 0103202 0103206 0103158 0103162 0103090 0103174 0103118 0103178 0103182 0103214 0103190 0103218 0103194 0103222 0103762 0103766 0103770 0103690 0103778 0103706 0103794 0103742 0103674 0103746
0103754
0103750 0103798 0103802 0103806
Average % Moisture
44.0 45.0 48.0 45.0 43.0 54.0 49.0 43.0 43.0 48.0 45.0 50.0 49.0 48.0 45.0 45.0 44.0 59.0 45.0 46.0 45.0 48.0 45.0 48.0 50.0
56.0
42.0 51.0 44.9 44.0
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Table LIII. Summary of Moisture Determinations for Quail Membrane Samples
Sponsor ID
E01-0259-20507 E01-0259-20509 E01-0259-20526 E01-0259-20738 E01-0259-20742 E01-0259-20745 E01-0259-20748 E01-0259-20750
Centre ID
0101991 0101999 0102067 0102915 0102931 0102943 0102955 0102963
Average % Moistui
45.0 65.0 65.0 60.0 60.0 68.3 70.0 68.3
The percent moisture was determined for eight out of the 30 samples received because there was not enough sample available for the other samples.
Centre A nalytical Laboratories, Inc.
Page 50 o f 102
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Table LIV. Summary of Moisture Determinations for Quail Albumen Samples
Sponsor
ID
E01-0259-20506 E01-0259-20507 E01-0259-20520 E01-0259-20521 E01-0259-20509 E01-0259-20526 E01-0259-20514 E01-0259-20527 E01-0259-20515 E01-0259-20528 E01-0259-20529 E01-0259-20531 E01-0259-20532 E01-0259-20518 E01-0259-20533 E01-0259-20735 E01-0259-20736 E01-0259-20737 E01-0259-20720 E01-0259-20738 E01-0259-20742 E01-0259-20743 E01-0259-20744 E01-0259-20745 E01-0259-20746
EO1-0259-20747
EO1-0259-20748 E01-0259-20749 E01-0259-20750 E01-0259-20751
Centre ID
0101988 0101992 0102044 0102048 0102000 0102068 0102020 0102072 0102024 0102076 0102080 0102088 0102092 0102036 0102096 0102904 0102908 0102912 0102844 0102916 0102932 0102936 0102940 0102944 0102948
0102952
0102956 0102960 0102964 0102968
Average % Moisture
87.0 79.0 89.0 88.0 85.2 87.0 88.0 86.0 89.0 87.0 89.0 88.0 88.0 87.0 87.0 88.0 86.0 87.0 87.0 87.0 86.0 82.0 84.0 87.0 86.0
89.0
86.0 85.0 87.0 86.0
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Page 51 o f 102
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Table LV. Summary of Moisture Determinations for Quail Yolk Samples
Sponsor
ID
E01-0259-20506 E01-0259-20507 E01-0259-20520 E01-0259-20521 E01-0259-20509 E01-0259-20526 E01-0259-20514 E01-0259-20527 E01-0259-20515 E01-0259-20528 E01-0259-20529 E01-0259-20531 E01-0259-20532 E01-0259-20518 E01-0259-20533 E01-0259-20735 E01-0259-20736 E01-0259-20737 E01-0259-20720 E01-0259-20738 E01-0259-20742 E01-0259-20743 E01-0259-20744 E01-0259-20745 E01-0259-20746
E01-0259-20747
E01-0259-20748 E01-0259-20749 E01-0259-20750 E01-0259-20751
Centre
ID
0101985 0101999 0102041 0102045 0101997 0102065 0102017 0102069 0102021 0102073 0102077 0102085 0102089 0102033 0102093 0102901 0102905 0102909 0102841 0102913 0102929 0102933 0102937 0102941 0102945
0102949
0102953 0102957 0102961 0102965
Average
% Moisture
48.0 43.0 46.0 45.0 45.0 49.0 49.0 47.0 54.0 47.0 47.0 41.0 49.0 50.0 47.0 47.0 46.0 47.0 41.0 43.0 44.0 43.0 45.0 53.0 46.0
47.0
46.0 45.0 45.0 43.0
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FIGURES
Centre A nalytical Laboratories, Inc.
Page 53 o f 102
Centre Study No.: 023-063
Figure 1. Typical Calibration Curve for PFOS
M
i 8
Centre A nalytical Laboratories, Inc.
Page 54 o f 102
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Figure 2. Chromatogram Representing a 0.1 ng/mL standard for PFOS
Saapla Index: Saapla Type:
Concentration: Aeq. Date: Acq. Tiae:
1 Standard
0.100 03/20/01
02:59:28 PM
Modified:
Bunehing Factor:
tfolae Threshold: Area Threshold: Mua. Saoothai XT Window:
Expected XT:
Sap. Width: Sap. Height: Exp. faak Katie: Exp. Adj. Katie:
Exp. Val. Katie:
Use Relative XT:
Yea
1 5.0
25.30 2
30.0
4.43 0.20 0.01 $.00 4.00
3.00 Wo
aae ain
5
i* 5
1
Xnt. Type: Xatantien Tiae:
Area: Haight: Start Tiae: End Tlaa:
Manual
4.44 ain 1478.3 count 3.5-oo: cpa
4.34 ain
4.St ain
312 350 300 250 200 150 100-
80
0
---
1.5 2.0
2.5
4.0 Tim,min
Centre A nalytical Laboratories, Inc.
Page 55 o f 102
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Figure 3. Chromatogram Representing a 1.0 ng/mL standard for PFOS
S ritN ^ Sttgetn ^lO r^jSw iW C eSi Pmk
Sas^le index: Saaple Type:
Concentration: Acq. Oate: Aeq. f i M i
*
Sta1n.0d0a0rd
03/26/01 03:37:1? PM
Modified: Bunching Factor: Noiso Threshold: Arts Threshold:
Nua. Saooths: RT Window: Expected RT:
Sep. Width: Sep. Height: Exp. Teak R e d o : Exp. Adj. Retioi
Exp. vel. Ratio: Use Relative OT-.
1
5.08
235.30
30.0
40..2403
0.01 5.00 4.00 3.00 Mo
Xat. Type: Reteaeion Tine: Area:
Height: Start Tistt: End Tine:
Base To Base 4.44 ain
7367.4 counts
l.?2e+003 cps 4.29 ain 4.61 ain
Centre A nalytical Laboratories, Inc.
Page 56 o f 102
Centre Study N o.: 023-063
Figure 4. Chromatogram Representing a Reagent Blank for PFOS (Centre ID Reagent Blank A, Set: 032601A)
S w o t * Induct Sanpl Typ*:
Concentration: Acq. Date:
Acq. Tin:
10
0l.0a0n0k
03/2S/O1 04:52:51 PH
Nodlfiodt Bunching Factor:
Mois Thrshold: Aroa Threshold:
Mun. Smooth: RT window: ExpCtd RT:
Sp. Width:
3p. Haight: Exp. Pak Ratio: Exp. A d j . Ratio:
Exp. Val. Ratio: 9 Relative RT:
Ya
1 5.0
25.30
2
30.0 4.43
0.20 0.01 5.00 4.00
3.00 MO
ae ala
1
e
J
Znt. Type: Rtntion Tin: Ara: Haight t Start Tin: End Tin:
Manual 4.45
490.3 8.S8+00J
4.37
4.58
in
counts cps
in nin
110 100
90
ao
70 90
so
40 30 20
10
0
445
Centre A nalytical Laboratories, Inc.
Page 57 o f 102
Centre Study No.: 023-063
Figure 5. Chromatogram Representing Control Mallard Membrane for PFOS (Centre ID: 0003802 Blank A, Set: 032601C)
gBMqaaSM
Staple Index: ~ Saapi Type:
Concentration* Acq. Date* Acq. Tiae*
12
Blank
0.000 03/27/01
03:10:07 PH
Modified:
lunching rector* Moioe Threshold: Ares Threshold: Mua. Saootftsr
XT Window: Expected JtT: Sep. Width: Sep. Height:
Exp. reek Ratio: Exp. Adj. Ratio: Exp. Vel. Ratio: Use Relative XT:
1Yes
9.0
29.30 2
30.0
4.43 0.20 0.01
9.00 4.00 3.00 No
Inc. Type:
Retention Tiae: Area: Heighc: Start Tiae:
End Tiae:
Manual
4.44 0(9.0 1.4(e*003
4.37
4.01 In
f9t g |
Centre A nalytical Laboratories, Inc.
Page 58 o f 102
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Figure 6. Chromatogram Representing Control Mallard Membrane Fortified with 10 ppb of PFOS, (Centre ID: 0003802 Spk A Dup. Inj., Set: 032601C)
C ntnw *3W B lsSei0q
Sanple Indexs
15
Saisie Types
0C
Concentracions
0.300
Aeq. Satot
03/27/01
Acq. Tin*i
03J516 PH
Modified*
Bunching Factors Koiao Threaholds
Area Threaholds Hua. Snootha:
AT Windows
Expected ATi Sap. Widths
Sap. Haight:
Sxp. Peak Aaeioi
Cxp. Adj. Ratios Exp. val. Ratios
Uae Ralativ RTs
Ye* 1 3.00
23.30
2 30.0
4.43 0.20 0.01
3.00 4.00 3.00
No
aee ain
Xnt. Typas Ratantion Tiaai Areas Haights
Start Tinas End Tinas
Manual
4.44 ain 1346.3 counta l.3e*002 epa
4.34 ain 4.61 ain
240 33
300'
280
240*
140
12
Centre Analytical Laboratories, Inc.
Page 59 o f 102
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Figure 7. Chromatogram Representing Mallard Membrane Sample from Week 1 at 6.2 ppm ai (E01-0263-20825) (Centre ID: 0103092, Set: 032601C)
Saapla Index: Saapla Typ*s
Concentration: Acq. Data: Acq. Tiaa:
30 Unknown
W/A 03/21/01 07:04:57 PM
Moditiad: Bunching Factort
Noiae Threahold:
Art* Threshold: Nua. Saoothai RT Windowt Expected RT
Sap. Width
Sap. Haight: Exp. Peak Ratio: Exp. Ad j . Ratio: Exp. Val. Ratio:
Uaa Ralativa RTs
ho
1 5.0<
25.30
2 30.0
4.43
0.20
0.01 5.00 4.00
3.00 VO
aac ain
4000
3900
3000
ftIT 250911 2000
Xnt. Typai Ratantion Tiaa: Araa:
Haight:
Start Tiaa: End Tiaa
Baaa To Baaa
4.43 ain 20013.1 counts 4.540+003 epa
4.29 ain 4.44 ain
1500 1000
SOS'
3J1.5 2.0 2.5 3.0
4.0 43 5.0 $.5 0.0 6.5 7.0
Tlma,inif
Centre A nalytical Laboratories, Inc.
Page 60 o f 102
Centre Study N o.: 023-063
APPENDIX A
Study Protocol 00P-023-063
(Centre Study No. 023-063) and
Amendments and Deviations
Centre A nalytical Laboratories, Inc.
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Centre Study N a : 023-063
CentreProtocol No. 00P-023-063
ANALYTICAL PHASE PROTOCOL
ANALYTICAL PHASE TITLE
extraction of potassium perfluorooctanesulfonate from egg
MEMBRANE, YOLKAND ALBUMENFORANALYSIS USINGHPLCELECTROSPRAY/MASS SPECTROMETRY
SPONSOR 3MEnvironmental Technology andSafetyServices
Building 2-3E-09 POBox 33331 St Paul, MN 55133-3331
DATA REQUIREMENTS Analytical MethodRequirements
PERFORMING LABORATORY
Centre Analytical Laboratories, Inc. (Centre) 3048 Research Drive
State College, PA 16801 Phone 814-231-8032
PROTOCOL IDENTIFICATIONNUMBER 00P-023-063
CentreAnalyticalLaboratories, Inc. Centre A nalytical Laboratories, Inc.
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Centre Study N o.: 023-063
Centre Protocol No. 00P-023-063
Title: EXTRACTION OF POTASSIUM PERFLUOROOCTANESULFONATE FROM
EGG MEMBRANE, YOLK AND ALBUMEN FOR ANALYSIS USING HPLC ELECTROSPRAY/MASS SPECTROMETRY
TABLE OF CONTENTS TABLE OFCONTENTS______________________
....
Page ,,
1. PURPOSE...................................................................
2. REFERENCEMATERIAI______________________
3. SPONSOR_____________________________ ____
4. TESTINGFACILITY... ........... .............. ..................... .........
5. PERFORMINGLABORATORY--------------------- ------
6. PROPOSEDEXPERIMENTALTIME-FRAME..... ......
7. JUSTIFICATIONFORTHESELECTIONOFTHETESTSYSTEM ... A 8. SAMPLEPROCESSING. STORAGEAND IDENTIFICATION
9. ANALYTICAT.METHOD-------- --------------- ------------
10. EXPERIMENTALDESIGN__________________ ____
11. PROTOCOLAMENDMENTS AND DEVIATIONS..................
12. RECORDS___________________________________ ___
13. QUALITYASSURANCE............-...........................................
14. DATA ANDREPORT.....................................................
15. archive statement.............................................. ,
16. REFERENCES.............. .......................... ............ ............
17. PROTOCOLAPPROVAL...................... ............................
APPENDIX: ANALYTICALMETHOD........................ ....... ...................... in
Centre Analytical Laboratories, Inc.
Centre A nalytical Laboratories, Inc.
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Centre Study N o.: 023-063
Centre Protocol No. OOP-023-063
EXTRACTION OF POTASSIUM PERFLUOROOCTANESULFONATE FROM EGG MEMBRANE, YOLK AND ALBUMEN FOR ANALYSIS USING HPLCELECTROSPRAY/MASS SPECTROMETRY
1. PURPOSE The purpose of this study is to analyze mallard and quail egg membrane, yolk, and albumen samples for residues of perfluorooctanesulfonate (PFOS) using method entitled, "Determination of Perfluorooctanesulfonate in Egg Membrane, Albumen, and Yolk by LC/MS/MS", Centre method number 0OM-O23-O15.
The analytical phase will be conducted under EPA TSCA Good Laboratory Practice standards 40 CFR Part 792 (1).
2. REFERENCE MATERIAL The following analytical standards will be used:
Test Materia] PFOS
TCR Number TCR00017-46
Purity (%) 97.9
Expiration Date 08/31/01
Chemical name and structure of the compound is presented below.
Chemical Name: IUPAC Name:
CAS Number Molecular Weight:
Perfluorooctanesulfonate
l-Octanesulfonicacid,l,UA33,4,4,5,5,6,6,7,7,8,8,8heptadecafluoro-, potassium salt 2795-39-3 499 (C8F,7S03">
O
II
C8F i 7S-- 0 `
II
0
Note: The neutral molecule and standard form that PFOS (anion) is derived from is perfluorooctanesulfonate potassium salt [CgFnSOjK], molecular weight 538. A record of test and reference substance receipt, storage conditions, and a record of use will be maintained at Centre Analytical Laboratories, Inc. Forms documenting chain-of-custody and shipping records for tracking of the test substances will be included as part of the raw data package.
All standards/test substances and any prepared solutions must be identified with a unique label or number on the container or cross-referenced to the container.
Centre Analytical Laboratories, Inc.
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Centre Study N o.: 023-063
Centre Protocol No. 00P-023-063
HAZARD INFORMATION
A current MSDS for the chemical(s) used in this study will be maintained at the testing facility.
3. SPONSOR
3M Environmental Technology and Safety Services Building 2-3E-09 PO Box 33331 St. Paul, MN 55133-3331
Sponsor Representative: Rochelle Robideau TELEPHONE: 651-778-7065 FAX NUMBER: 651-778-6176
4. TESTING FACILITY
Wildlife International, Ltd. 8598 Commerce Drive Easton, MD 21601
STUDY DIRECTOR: Sean Gallagher TELEPHONE: 410-822-8600 FAX NUMBER: 410-822-0632
5. PERFORMING LABORATORY
Centre Analytical Laboratories, Inc. (Centre) 3048 Research Drive State College, PA 16801
PRINCIPALINVESTIGATOR: Emily R. Stauffer, Centre TELEPHONE: 814-231-8032 FAX NUMBER: 814-231-1580
6. PROPOSED EXPERIMENTAL TIME-FRAME
Analytical Start Date Analytical Termination Date Report Issued
Mar. 19,2001 Mar. 31,2001
April 1,2001
7. JUSTIFICATION FOR THE SELECTION OF THE TEST SYSTEM
Mallard and quail egg membrane, yolk, and albumen samples collected by Wildlife International, Ltd. will be used as the analytical samples for this phase. Mallards and quails represent wild bird populations and they are an EPA recommended species because they do well in a laboratory environment. Quail
Centre A nalytical Laboratories, Inc.
Page 4 of 38
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Centre Study N o.: 023-063
CentreProtocol No. OOP-023-063
habitats include bushy pastures, grassy roadsides, farmlands and open woodlands
and they are considered land grazers. Mallard habitats include ponds, law-; and
marshes, and they are considered aquatic grazers. The use of these matrices for
the study was to determine if PFOS was deposited in the egg membrane, yolk, and
albumen of mating adults that were fed PFOS.
'
A complete description of the test system along with all of the in-life parameters is documented in Mallard Protocol No. 454/010600/MP/SUB454, Project No.
454-105 and Quail Protocol No. 454/01060Q/QP/SUB454, Project No. 454-104. This project is the analytical portion of those studies.
8. SAMPLE PROCESSING, STORAGE AND IDENTIFICATION No processing will be required for the any of the samples.
Each sample will be assigned a unique sample identification number at Centre
which will be used for tracking and identification of the samples. The samples
will be stored in a temperature-monitored freezer, maintained at < -10 C, except
when removed for extraction and analysis as described in the
The
samples will be kept isolated from the test substance during storage.
Sample receipt and storage location and conditions during the study will be documented. All samples and any resulting sample extracts will be identified with a unique label or sample number. Such identification will be either on the container or cross-referenced to the container.
9. ANALYTICAL METHOD
All samples will be analyzed according to the analytical method titled
"Determination of Perfluorooctanesulfonate in Egg Membrane, Albumen, and
Yolk by LC/MS/MS", Centre method number 0OM-O23-O15
'
10. EXPERIMENTALDESIGN
Samples obtained by Wildlife International Ltd. have been shipped to Centre Analytical Laboratories, Inc. for analysis. The are no known contaminants expected in the test system that would affect the results of this study.
Only those samples designated by the Sponsor will be extracted and analyzed at Centre according to method, "Determination of Perfluorooctanesulfonate in Egg Membrane, Albumen, and Yolk by LC/MS/MS", Centre method number 00M023-015.
Methods to control bias will include assay of untreated control samples, fortification of untreated control samples to obtain recovery data, and replicate
Centre Analytical Laboratories,Inc. Centre A nalytical Laboratories, Inc.
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Centre-Study N o.: 023-063
Centre Protocol No. 00P-023-063
analysis of fortified samples to provide an indication of reproducibility. Fortification will be made to the matrix prior to extraction at levels ranging from the LOQ to above the highest residue found. The fortification solution containing the test/reference substance will be applied to the designated samples via a Hamilton syringe, micropipette, pipet, or equivalent, to maintain consistency and accuracy.
The average recovery and relative standard deviation of the fortified samples will be calculated. Also, the average residue found and standard deviation for each matrix will be calculated.
The percent moisture will also be determined for each extracted sample if adequate sample remains after extraction. For each matrix, 0.1 g samples will be weighed and dried in an oven at - 105C for - 12-14 hours. Percent moisture for each sample will be determined in duplicate when there is sufficient sample available.
11. PROTOCOL AMENDMENTS AND DEVIATIONS
Any deviations from the protocol or from the analytical method as provided will be documented and reported promptly to the Study Director and Sponsor Representative. Planned changes to this protocol or to the analytical method will be made in writing as an amendment and approved by the Principal Investigator, the Study Director, and the Sponsor Representative. Any amendments or deviations will be appended to this protocol and included in the final report.
1. Pmtnral amendments: Planned changes to the approved protocol shall be Hnfuiwntd by amendments that clearly describe the change, justification for the change, and impact on the study. Amendments will be signed and dated by the Principal Investigator, Study Director, and Sponsor Representative. Copies of amendments will be sent to the quality assurance unit.
2. Protocol deviations: Protocol deviations, which are one time and unplanned deviations from the protocol, shall be documented in the study records, noting the nature of the deviation, potential effect or impact on the study, and corrective action if required. Protocol deviations are signed and dated by the Principal Investigator, Study Director, and Sponsor Representative and reviewed by QAU.
12. RECORDS
Records to be maintained include, but are not limited the following (as appropriate):
1. Sample tracking sheet(s) 2. Sample receipt records, storage history, and chains of custody 3. History and preparation of standards (stock, fortification, calibration)
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Centre Study No;: 023-063
Centre Protocol No. OOP-023-063
4. Description of any modifications to the method 5. Instrument run sheets, bench-sheets or logs 6. Analytical data tables 7. All chromatographic and instrumental conditions 8. Sample extraction and analysis dates 9. A complete listing of study personnel, signatures and initials 10. Chronological presentation of all study correspondence 11. Any other data necessary for the reconstruction of the study
All chromatograms will contain the following:
a. Sample identification, date, arrow or other indication of the area of interest, and injection number corresponding to the run.
b. Additionally, fortifications will include the fortification level of the analyte.
c. Analytical standard chromatograms will additionally include the concentration (e.g., pg/ml, ng/mL, ppb, ppt, etc.).
Each data set will contain information on temperatures, flow rates, column parameters, gases, instrument parameters, and instrument type, etc. The Centre study number will also be recorded on the first chromatogram of each daily analytical run.
13. QUALITY ASSURANCE
Centre QA Unit will review the protocol, audit the study conduct and the study documentation (including raw data and final report) to ascertain that all GLP standards are met Centre QA Unit will inspect the study at intervals adequate to assure compliance to GLP's, and will report the findings of the audits to the Principal Investigator, Centre Management, and the Study Director.
14. DATA ANDREPORT
1. All raw data and the original signed protocol will be maintained in the study file. This data includes the protocol amendments, protocol deviations, laboratory notebooks, analytical standard solution preparation, sample chain of custody sheets, sample work sheets, chromatograms, calibration curves, and any other appropriate data generated. Raw data not used will be stored in the study file. The reason for exclusion will be documented.
2. A report will be issued by Centre, approved and signed by the Study Director and the Sponsor Representative and sufficient for submission to EPA. The report contents should include, but not limited to: 1. Objectives and procedures stated in the protocol 2. Analytical and statistical methods used
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Centre Study N o.: 023-063
Centre Protocol No. OOP-023-063
3. Reference materials identified by name, lot, purity, and other characteristics
4. Name ofperforming laboratory and analytical start and termination Hat^c 5. Statement prepared and signed by the quality assurance unit 6. Tables containing all applicable data 7. All chromatographic and instrumental conditions 8. A complete listing of Centre study personnel
Centre will send a copy of the draft report to the Study Director and Sponsor Representative, who will return the report with comments. Centre Analytical will make revisions and finalixe the report with the approval of the Study Director and Sponsor Representative. The Centre QA unit will conduct an audit of the final draft report and data files to assure accuracy and GUP compliance. The report must be approved by the Study Director and Sponsor Representative prior to signature. A statement of QAU inspections and a statement of GLP compliance will be included in the final report
Any corrections or additions to the final report shall be in the form of an amendment by the Principal Investigator. The amendment shall clearly identify the part of the final report that is being corrected and the justification for correction. The amendment shall be signed and dated by the Principal Investigator, Study Director and SponsorRepresentative.
REPORT DISTRIBUTION: Original: Study Director Copy: Centre Archives
15. ARCHIVE STATEMENT
Study records to be maintained: Records to be maintained for the study include all raw data, observations recorded during the conduct of the study, documentation, chromatograms, specimen tracking information, and study related correspondence. This includes a description of equipment used during the conduct of the study. All characterization data and any shipping records shall be retained.
Document archives: Upon completion of the study, the study records, protocol and amendments, and the final report and amendments shall be retained in the 3M document archives. If it is necessary to substitute a copy for an original record, it will be certified as an exact copy. Centre will retain facility-related originals and a copy of the final report and raw data package.
16. REFERENCES
1 U.S. Environmental Protection Agency. Toxic Substances Control Act (TSCA). Good Laboratory Practice Standards. Final Rule, 40 CFR Part 792.
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Centre Study N o.: 023-063
Centre Protocol No. OOP-023-063
17. PROTOCOL APPROVAL This protocol was audited by the Quality Assurance Unit of Centre Analytical Laboratories, Inc. Quality Assurance Reviewer, Centre
_______ uX Naomi Lovalio, PhD. Quality Assurance Auditor
PrincipalInvestigator, Centre
3 /Wt?/ bate
Facility Management, Centre
q L j jU ^ I -
yioh n Flaherty 7 / Operations Manager
S tudy D irecto r, W ildlife In tern atio n al, L td .
A L l JMLl ,,
Sean Gallagher
'
Senior Biologist
Sponsor, 3M
Date
I h /o I
Date
Centre Analytical Laboratories, Inc.
Centre A nalytical Laboratories, Inc.
Page 9 o f 38 Page 70 o f 102
Centre Study N o.: 023-063
Centre Protocol No. 00P-O23-063
APPENDIX: ANALYTICAL METHOD
A. "Determination of Perfluorooctanesulfonate in Egg Membrane, Albumen and
i Yolk by LC/MS/MS", Centre method number 00M-023-D15
'
Centre Analytical Laboratories, lac:
Centre A nalytical Laboratories, Inc.
Page 10 of 38 Page 71 o f 102
Centre Study N o,: 023-063
CentreProtocol No. 00P-02363
.
. TITLE ,
:
Determination ofPerfluoroocanesulfon*a"t*e*inUEggMembrane, Albumen
LQMS/MS
Wd Yolk by
AUTHORS EmilyStaufferandJohn Flaherty ' .
DATE ISSUED ' " January2,2001
. SPONSOR .
3MEnvironmentalLaboratory ; ' . Buding 2-3E-09 . POBox33331 '
` ' Sl Pau!,MN53133-3331
PERFORiVflNGLABORATORY
3048ResearchDrive . State CbDege,PA 16801 '
CENTRE STUDY NUMBER
023-015
: CENTRE METHOD NUMBER
"I
0OM-023-0I5
. *
. TOTAL NUMBER OF PAPTC
Centre Analytical Laboratories,'Inc' Centre A nalytical Laboratories, Inc.
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Centre Study N o.: 023-063
CentreProtocolNaWP-023-063: ' '
' Centre Method No: 00M4&M5
. . MANAGEMENTAPPROVAL.
'
. As per40 CFR 792J, methoddevelopment is not required to be conducted'
with GoodLaboratory Practices. However, the work was in conformance 1.n,con3Pi" nce
standard operating procedures andgeneral GLP regulations.
Wlth applicable
4 Q 4 io l
_ J S tanffer
Date
Principal Investigator- ` " '
CentreAnalytical Laboratories,hie.
-Q L
iM f
./JohnHaheny * " * ' Date
laboratory Manager .
*
Centre AnalyticalLaboratories,Inc.- _
AM
/*
I1 - iU fl- O
^Qrfcim. PhD. Prerideat.'
Date
Centre Analytical Laboratories,Lie. '
" r yyD^e.
/
Caere Analytical Uboretories.Inc.Study9.0Q3-O1S
' Centre Analytical Laboratories, iri" Centre A nalytical Laboratories, Inc.
Page2 '
Page 12 o f 38 Page 73 o f 102
Centre Study N o.: 023-063
Centre Protocol No. OOP-023-063
TABLE OF CONTENTS
TITLE PAGE------------- -- m anagem ent PPROVAL.
-1
.2
TA?LE OF CONTENTS LIST OFTABLES_____
.3
t
.4
UST OFFIGURES.
-
L SUMMARY-
-i.
2. ex perim en ta l com pounds.
-7
3. CHEMICALSAND SUPPLIES --
3.L CxatEAU--
3i S a M o a ss.
33.qamoeirANDSumi&r
O
3.A Saunons
33. he**AMwSTOCX.ftinff*aT*A>eCAUWaTMSouniis.
3JJ. Sud Sabt*.
3S Z FonificanoitSolutions--
3 S 1 Colraion Standards--
< METHOD..----
4*1. FunrOufiUM__-- 43. SamrsFujcssnS-
4 3 . SAMRSBCnCin*-
<A1EgMtairBU.
4- EjjK)Saa4iUiwKi(
4.4. AMK AuUsTsSi Br Il CY UUC ISMMS MSS -- -------------
-- ----
- 4.4.1..LCMS/HSSystm and O fim tinf Conditions(Ttrpolonspriy)-
t . 4.412.' Fmmpl*Xu FtU Parmnetas---- ---- -- -- ----
'4.4J. CaSraim fn es n ra . ^ *
................ - 1 .
4.4.4. SmpUAaafyrit........... --
'" . .
43. FatnxiMMCsOBExa------- ;------- :-------------
4.6. T1MERB5QBS)TO*AfOLYIB-- ---------------------------------
s. .CALCULATIONS-- ------ ;------- r ---- -
.10
-10 -10 -10 -11
-U
-1 2 -12 -IS -13 -1 4
-15
-16
.16
SAFETY_
.17
TABLS____
.18
FIGURES.
"n
CsaBe A m ljoalU bcniorie. Ibc. Sm4j 4,0234113
fcr-3
. Centre Analytical Laboratories, Ine." Centre A nalytical Laboratories, Ine.
'.
. ' ' Page 13 o f 38 " P a g e7 4 o fl 2
...............Centre Study N o.: 023-063
: CentreProtocolNo. 00P-023-063
. CentreMethodNo: OOM-023-015
LIST OF TABLES '
'
ta ble i: summary o f recoveries for pfos in quail egg' . ` ` MEMBRANE--:---------------- :-----------_______ _, '
ta ble il- summary o f recoveries for pfos in mallard egg
` ; MEMBRANE----------:---------- >-------------
`
-19
'
TABLEn t SUKftiARYOFRECOVERIESFORPFOS IN QUAILEGG ' '
' YOLK_i__ _______________________ --
.20
Ta b l e iv : summary ofrecoveries for pfos in Mallard egg
. ' YOLK------------------;---------------- ________________ _ --20-
TABLE V: SUMMARYOFRECOVERIESFORPFOS IN QUAILEGG ' ; .' ALBUMEN-----------------------------:------------- '
--.21
TABLE V t SUMMARYOFRECOVERIESFORPFOSIN MALLARDEGG
` ' ALBUMEN.---------
. -21
C aars Analytic! Labonario. Ire. Sindy S02M U
Centre Analytical Laboratories, Inc. Centre A nalytical Laboratories, Inc.
Pegs 4
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Centre Study N o.: 023-063
Centre ProtocolNo. OOP-023-063
CentreMethod No: ltfM.023.0j5
LIST OFFIGURES-
fisw e 1: Representative Chromatogram ofa QuailE y MembraneComnii
-23
figure 2: Representative Chromatogram of1 MallardEggMembran; Control
, .*
I
' figure 3: Representative Chromatogram ofa QuailHy YolkControl.
-23 .2 4
. ' figure 4: Representative Chromatogramofa MallardEggYolkControl.
.2 4
Kgnre 5: Representative Chromatogramofa QomTEyAlbumen Control
1-25
figure 6: Representative Chromatogram ofa MallardEggAibunren Control
figure 7: -RepresentativeChromatogramofa QuailEggMembraneControl~
' . . lOng/gTppb)-- ------------;_________ ___
^
-.2 5
-r *
"edat 26
figure 8: Reorcsentative Chromatogram ofa MallardEggMembrane Control p-_-r
- 10.0 ng/g (ppb)__------- :--------------:--
tt?uCed at -- 26
Hgure 9f Renrcsentative Chromatogramofa-Quai]EggYolk Control Fortifier) t "
. ' 10.0 ng/g (ppb)------------------------ ___________ _
-' -2 7
' HgunrlO: Representative ChromatogramofaMallardEgg YolkControlFortified
- ; ` . 10.ng/g (ppb):----------- ----~ -- --
* "
figure.ll; Representative Chromatogramofa Qna EggAlbum Control n _ ` 10.0 ng/g (ppb)------------------- :------- j-------------
' figure 12: Representative Chromatogtamofa MallardEsg Albumen Control Forti'f H : ' . 10.0 ng/g (ppb)------------- -- -- --1-------------- --. le^
C=areAnalyticalLaboratories.I*. Study*023-015
Centre nducdTj&ratonsTInc. Centre A nalytical Laboratories, Inc.
for-5
Page 15 o f 38 Page 76 o f 102
Centre Study No.: 023-063
_ CentreProtocol No. OOP-023-063
I " trt Method No: OOM-Oa^tf
L SUMMARY '
.
''
'
Hus docatneat details a method of analysis for the residual
Ptfluorooctanesulfonate (PFOS) in egg membrane, albumen and jo lt I ? Inall'on of .
formula of the"analyte'is.given in Section 2 of this method
Hie chemical
' '
Perfluorooctanesulfonate is extracted from each matrix with methanol ftfenm
tuonibtane samples, the methanol extra is passed thrash a
^r egg
alhrnnm '.j --m ___ i-- . n c . - c
. . ",s" ane filter.- R*- --
SexBtpraj'clmt. iMQui-a.n/Tti/f*icna/tioonn ofP-F_OiS_i;s_a_cc_o_m_plii_sh_e_dr`b`y li.quid dm xn.awt5o*m"iJfjr/t3l](oejgejntf,anl
For this method the LOQ is 10 ngfg (parts-per-bilh'on). -This j
-
conducted with egg membrane, albumen, and yolk control samni-- y*1. 00 ruddies
development.'
'- .
-T
pIes durinS method
C=r: Xmtyiical Labomoriej, Inc. Studyi C M IJ
Centre Analytical Laboratories, Inc. Centre Analytical. Laboratories, Inc.
Pajs6
' ` Page 16 of 38 . '
' Page 77 of 102
Centre Study N o.: 023-063
, CentreProtocol No. OOP-023-063
Centre Method No: 0OM-O23^is
EXPERIMENTAL COMPOUNDS
The m oleste antenne ofPFOS is given below: .
PFOS . .
Molecularweieht: 499 (CjFnSOj- ) - .
''
*.
o' '
. -s
o
.
- . *.
`
ChemicalNic e : Perfluorooctanesulfonate
'-
IUPACnans :l-Ocqnesulfonic icid,U 22J,4,4Jj,6,6,7,7t8 g.
'
hepiadcafluoro-potassiumsalt ' :
''
CAS#:
2795-39-3
`' ' '
` Note:
fromB^perfluorooctanesulfonarepotassium salt [CirnSOiKl '(
'
' .w s tS S S ) .
uiar
'3.-. CHEMICS-ADSUPPLIES. 3.i: CHBflCALS
-
Chemical______
Can " Methanol (MeOH)
_ Ammonium Acaate ' Water
Grade
120/400 HPfcC Reagent Tvoel
;
Source
.i ' Calalo Nn
Supdco . .
` 57210-U
(YWR)J..r Baker
Aldrich
*
Centre "
.
JT9093-2 11X-7-5
"
' (TypeI water=electrical resistivity, minimumof 16.61 Mit-cm at 25r f"-----
` Labcooco1" waterptoworkstation, based'oa ASTMclassification)
rm 1
' 3.2. Standards .
Standard Grade PFOS _ Analytical
Test Control Reference Number
TCR-000I7-4
^MfeivjroiuneataT
UboratoO'.St.PauiiMN
.
Centre AnalyticalT.
be*3tody9 023415
* i,
hje7. '
Centre AnlytcaE&oratdnerInc. Centre A nalytical Laboratories, Inc.
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Centre Study N o.: 023-063
CentreProtocofci' 0P-23-063'
.33. Equipmentand Supples
Cintre Method No: MM.023.oij
EQUIPMENT
Balance, 5 place analytical
Balance, 1place Uploading
MiniBeadBeater-8 - '
Wrist-action shaker
-
BendHop centrifuge
Mini-centrifuge-1201 *
50 mLdisposablepolypropylene centrifuge'tubes
Stainlesssteel beads (catf 11079)
'.
Mjcnxcntrifuge tubes *
.
Disposablepipettes,testrobes etc.
15-mLdisposablepolypropylene centrifuge tubes
.WhatmanTMAnocopTMfiller (cat# 6809-1022)
. 2-mLEpoendorfTMimcrocentrifuge tubes (cat?22
36-3353)
'
MettleMettle
BioSpecftnducts Butte]] EC VWR VWR
Bio sI*c Products VWR VWR
' VWR . 'VWR' VWR '
2-mLdearHPLCvialbit (cat i 5181-3400) . Standatd.lab equipment(dass A pipettes'and
' HP
volumetric flash, gratitraM cylinders, etc.)
LC/MS/MS andHPLCsystems . . - ( A*describedin
Section4a i
'N ote:
L In Older to avoid contamination, the use of
y** .
..highlyrecommended (containers, tubes, pipettes, erojr- TMwarc
- ' Z Teflon or tefloolmed containers or eqiripment/incJudm t.n -
HPLCvialcaps,should not be used. . *
ton-lined
; ' 4. It is neessaty to cheek the solvents (methanol) fir
-
contaminants by LC/MS/MS before use. Cenain ]
been foundto heunsuitable foruse. .
number have
' 5. Use disposable micropipettes or pipenes to aliquot standard t '
andwhenpreptnng'standards and samples forextraction. ' SOUOns
" 6. .Equivalent matenals may be.substituted for those suerif^i ' .
method Bowver.'lhe use'of carbon flom j , . : . " .ds
irmrnmnnfcd *
.
' 8 ^ g ly
3.4.'Solutions. .
.
' '
1. 50 tnM ammonium acetate solution:' Dissolve 3.J5 ,,#
.* `
acetate in HofASTM typeI water. Store in an ap p ro j^.
aroom teapenmttfbruptolyear.
. ^ooule
. 1 ' 2 mM ammonium acetate solution: Dilute 40 mL of the so
ammonium aceate solution in a liter of ASTM type I
f a M
C-c Analytical Lahon io tic . Is . Sftaty 1023-015
K8
Ceafre Analytical iibratoiftesrliiE Centre A nalytical Laboratories, Inc.
Page 18 of 3 8 . Page 79.o f 102
Centre Study N o.: 023-063
: Centre Protocol No. OP-023-063
: ' ; : . Centre Method NM 0M -023^5 /
ipoha1s-eyeAar.. Store in an appropriate glass kittleat room^tePam^ t-ui re f,,or up
.
Note
Toe volumes shown may be prepared
are
provided '
for
dance;
aliena' ,ve
volumes
.3.5. Preparation of Stock, Fortification, and CaL b
'
. Solutions - .
. r BRA'n0N
1;
'Analytical standards.are prepared for two purpose^. Hey are ii^
entreated samples in order to determine analytical recovery and too i"
. responseofthe detectornsedin themalysis. . '
Wuorateth-
'
The.analystmayvary theabsolute volumes of the standards as Ion" k proportions of solute to sofanfare maintained. The solutions died iJ COiIia ' givenasaue^mple; alternativeconcentrationsmay bepreparedifneeded -W**
.
* 3SL SlockSolution' r .
.
Prepare stock solution rf PFOS;at 100 ftg/mL.by weigBag out ,nn
analytical-standard (correctedforpertrentsalt). Adjust fiiiai volume f
. with methanol in a lOCMnL.volumetric flask. .Store this stock ? . mL
125-mLLDPHbottles) in a refrigerator at 2*C to 6'C for a maxim - ^ -n
. of6 monthsfitonrthedateofpreparation.
. ..n period'
. '
3S1 FortificationSolutions
. .
,
a. 1,0ug/mLFottiticarion Soution - Pipette 1.0 mT nf fa 1QQ - ' -johnon into a 100 ml/volumetric flask. Brin- im / `f mLst0<* .
" "P 10.-volume with
k -0 1 ;.;mL Fbmnation Solution - Pipette inn - 'i of d i i il . . .
focriflcarion soluti into a 100-mL volumetric flask
Tohnnewith methanol. ' .'
" * * * "P
: J '
*
c.`0.01 ntftnL FoitincaioB Solution - Pipette'inn ^ ^
.'
bonification solution into a 100-mL volumetric flask
1
. '.
volumewith methanol "
'.
m
up to
..
(
Store-til fortification standard solutions (in 125-mL LDPE botti 1
'
remgeatorat 2*Cto6 ^ foramaximumperiod of6months fiomScL
>
. 3 S X m iration Standards
, .' '
.. Prepare'sii LC/MS/MS alibtation standards'in methanol via dii.,-- , . .
0.1pg/mLand 0.01 pgfaL fortification solutions.
. of the
- .Centre AnaMed laloaorics, Inc. Study5 023-015 - "
. P. a*-0
Centre Analytical Laboratories, Ioc.'"
'
Centre A nalytical Laboratories, Inc.
~ ~~"epage-jg jg Page 80 o f 102
Centre Study No.: 023-063
Centre Protocol No. OOP-023-063 CentreMelh^a N:OOM-023-015 '
0.1
A1 ' 0.01 0D1 0.01
25 - 100
. 1`
100
.. 5 ;
- 100
1
. 100
- 05 li ,'
.
oar
0.002 0.001 ` 0.0005 : 0.0601 0.00005
. `Store all calibration standard solutions (in `125-inL LDPE bant \
' refrigeratorat 2`C to 6"C fora maximumperiod of6 months from dtl" * . .preparation. . . " dateof
4. METHOD
`
..
.
.4.1.' flowDiagram
-.
; The flow diagram of the.,method is given below,'followed hv , j .
description ofeachstep.
.
-. . ~ailed
. ' _ W eighlg ofmatrix (0.-lgformembrane).Fniiyif needed-"
".
. . i' *
-
*.
: "
` . . net with MeOH * *
-
'
.1 ' ' `
`" .
Carbon cleanup (exceptmembrane)
.
_ .
.
.
:`
*
.
.
. '.
. Hlter
: i .
'
'
.. Dilution
'
."
. -. 1 ` ' '. *
' ` LC/MS/MS anaiysis . .
' .
.
' 42. Sam ple Processing
. : ...
` ; All.samples are received freren andwill be kept froztn'(below -10 *q
ofth extraction.
.
, Ume
43-.. Sample Em -^cnoN . .
`
NOTE All eggsample matrices wereseparated priorto arrival at Centre.
.CsatreAnalytics!Labonscrics. Jos.Soidy 023*015
Pas10
Centre Analyticallaboratories,ifcl"" Centre A nalytical Laboratories, Inc.
"Page 20 of 38 Page 81 o f 102
Centre Study N o.: 023-063
Cintre Protocol Na 0P-23-063
1 Centre Method No: OOM-OZj^ '
4 3 .1 Egg M embrane
, "'
a. Allow sample to thaw.
.
b- Weigh OJ g ( 0.005) ofsample into a 2-mLP TM - . , ,
'
the weightto the nearest O.OOOlg. Fortify untreated control W* ' ^cord point fordetermination of method recovery. . ; saoPlc at this-'
Aaa i mum yicua ana ones smniess*stcsl beads, reblar* rj -* homogenize with bead beaterset atposition 7 (_250 0 mml 5" "SMyand
' Centrifuge the tubes with mini-cenntirriiffuugse at V X 0 r-o m 'r-*>Io0r5*" "t"utes."
' .th..e..n...u..i.c..i.i.i.l.f..y...t.r.a..n..s.f'er^!P--a2-nnaitnan*mtomaa new 2-mr c filtering withWhatmanTMAtntortonpoTM 10filterif necessar^y pgutorf^ tube..
d. . Make a JX (Slurion by transferring 0.1 ml, offfltrate to W r adtfing0.4 mLofmethanol rising disposable micropipeaes (10a20C^.^Kl
e. Store the tea of the extras In a refrigerator at approximately f n '
-forfuturere-dfiurion orre-injection.
cm 5*c-.
4 3 3 .. E gg Yolk and A lbum en
a.' Allow sampleto thaw.
"
' . :
b. Weigh 1.0 g fe 0.03g) of sample Into 50-mL (5 <TM ;,b|c nnll
'-
centrifuge tube: Record the weight to the n e ^ t 00001PtOP3<!^ i '
untreated control samples at this point -for -detenriinaiinn *V Fotsi6 ' ' :
recovery.
'
:
'-
* hod '
'` " ' .
.
c. -Add 25 mL of MeOH, replace lid tightly, and shake on a
.
shakerfor 15amnues. Centrifuge tubes i t - 2,000 iprn for- 1 0 ^ ^ ''" j
Transfer about 5 mL of extra into a 15-mL disposable ' t centrifuge mb* addO Jg carbon, replace lid tightly, and shake about 10seconds.
. ' `
e. L the tubes sit for--2 minutes, carefully tnmsfer-2.0 mL f'k
into a 10-ml'disposable syringe. barrel connected to a Wt, *Bapls
' AnotopTM 10 filter: Filter sample into two HPLC vials, one , '! annann<
. 'and one forstorage in a refrigentor at approximately o*r r .~ rU,je::ion
re-dnurionorre-injecaon. . . `
' " >rfuture
CentreAnalyticalUboranris.Inc.Studyi 0ZWJL5 '
'
' :' ` '
"' 'Fsgell
Centre Analytical Laboratoriesvinc:' "CentreA nalytical Laboratories, Inc.
Page21 of 38 Page 82 o f 102
Centre Study No.: 023-063
: CentreProtocol No. 00P^023^63
4.4. Analysis by LC/MS/MS
Centre Method No: OOM-023-015
4 .4 J: LC O S/M S System and Operating C onditions (T w io Iorup j
Mass Spec . PESC E API3000BiomolecularMass Aial . .
Interface
SOE X Tutholon SprayLiquid IntroductionW -f '
Harvardinfusionpomp ' ;
" aacs
PowerMacintosh G3
' Software
PESoexAiralyst L IWindowsNT
KPLC:
EewiettPadani (HPJSenes 1100 .
'
'' ' '
HPQaaiPnmp. ;
_j
'.
HP VacuumDegasser * '
.
- - ' HPAmosamnkr .
.. `
' ' . . HPCoTumnOvcn'.
-HPLCColumn: Genesis Ci (JonesChromatozraDhvl ) | ; .
ColumnTempj 35" C
- . *"
Injection VoL;... 10(iL '
* Mobile-Phase (A): 2 mMAmmoniumAceate in ASTMimi r
' MobilePhase(B): Methanol
`` .
-^
. How Rate: . OJmL/min.
.
Time
' 0 " U) . 7J)
7J
ILO
A
a
o o ao
40
S
40
100
100
` 60
60 :
'
. .
.` It may be oeeasaiy to .adjust the HPLC gradient in order to' / _instrument performance. Columns with different dimensions (e.* -?Umizs
' 30 mm) and columns fromdifferentmanufactures (Keystone Betasilr-mm * can beused,providedequivalentchromatographyis obtained ' . ^
Ions monitored: '
AnaNfe. _ Mode Transition Moniion-f)
PFOS. negative
499 ^ 99
Approximate
etentinn-p~-fnl||1
47q
CansAnalytical Laboratories. In5BtiySQ23*dLS
Page12 '
Cntre Analytical Laboratories;Inc.'
Centre Analytical Laboratories, Inc:
Page22 of3g
Page 83 of 102
Centre Study N o.: 023-063
pntre ProtocolNo. OOP-023-063
CentreMethod No: O O M ^o ^
4 .4 2 . E xam ple Tune F ile P aram eters * '
'
The following values are provided as an example. Actual val
from insuument to instrument. Also, these values may be Chan'S va,y
to rimein orderto optimize forgreatest sensiu'vhy.
snaaS*d from time
r.
.
The mass'spearometeris:tuned using a OJpgfrnLPKlS solution dQudon of the stock solution in methanol The soiyticn is *T" connector) at 10 jtLfrmn into a 0.2 mlAnin stream e
' vv*ia .a
consisting of40% methanol and 60% 2.mM antmtauum acetate.1? ,! ? ' piase
are initially timed for theparent ion and1then nmed ftoor the prod .. ^Wy tlyetess
the insmnnent is tuned, the optimized parameter are savedas i*" ^ nci
`This,tone fDeis then used during routine analysis. The tuning J L m 1* e"
be repeated,as necessaryto'ensure optimal sensitivity.
S Procedure jy
Canfrols
''
IS-Iospray.
"
OR-Otifice . - .
RNG-FocusRing _
QfcQuad.ORod Offset
. IQ-Interqu5d 1 lens '
_
'ST-5nibbi'
'
ROl-Quadl Rod Offset _
*
IQ2-hueruad21ens ' '
R02-Quad2 rod offset -
ST3-Smbbies .
R03-'.Qnad3 rod offset
DF-EMDefkction Plate
CEM-Oannel Bectron Multiplier
' G as Flows
NebulizerGa . . CmtainGas Collision Gas
TE T em perem re
Sst 4000.0
-61.0
-270.0
10.0
' 92
' ' 15.0
R3
20.0
" 84.0
` 10.0
86.0
300.0
2400.0
. Set
M2
13 '
4
350*C '
.
.
.
4 .4 3 . Calibration Procedures-
I*
'
*'
a. 'Inject the same volume (between 10 to 20 pL) feach calibration
(preoaredin MeOE) into the LC7MS/MS.* . .
standard .
Use linear. 1/x weighted standard curves for qnmrtft.-,,, ^ j . standatd curves are generated for each set'by finear regression usinv-'tte
Centre Aiwlyrii-,1t .hnrwww Tue,SmrfySOELittlS
Page 13
TXSHtR'Allalytical Labutatuties, tc."" Centre Analytical Laboratories, Inc.
-- .Page 23 of38
i
' Page 84 o f 102
Centre Study No.: 023-063
Centre Prtgocol So. 00P-23-O63
- : C" 1TM N:
aporopriate'software system. Anyeafibtation
_
, 305. based on its calculated conceatration, mm be esdriS atsi<,i -
afibration curve. However, the total numberofcaiH satint* *"*a ***
- . may be excluded must not exceed 30* of the t o t a l aa8^an^ that.
injected.. _
' . standards
c. The correlation coefficient ft) for calibration eaves r-n.i_._i
20.9925 (fS&SSS). If catibratiOT.'results fall ootade^fetL TM **
appropriate steps should be taken to adjust instrument cu ^ ._ ^ ^
relevantset ofsanqrles mustbereanalyzed.
?P'-*uon, and the
a. Ihjea the same volume used for the calibration standards o__ 25pL) ofeach sample, fortification,control, etc. into the
b. Standards corresponding to at least six concentrations
an analyticalset.
' 10
c. Inject an entire set ofstandards (six) arthe beginning oftbe-nm `
' standards hrterppKsed abotrt every-5-10 samples.. AD
^ "?= t
post be bracketedbystandard injections (see Section4
-, ea`ons `- ';
d. Each set.ofSamples analyzed (aot to exceed 25)nnrst include 'w .
reagent control (method blank), one ASTM Typei w ater-yank* '
one matrix control, and two matrix control a
a l" 8
concentrations andcarried through theprocedure toverifyrecovery ^nWn '
e. AUsamplesmust beanalyzedwith dupficate injections.
:'
' Note: The."analysis performed -during method *-- __ . . . fortificationsat 10,50 and 250 ngfgfppb) for the analyse. m" ud=d '
from tire-standard carve based orr the peak area of the jn^ 7 * `funed
standards injected during a se t .` The standard responses-m um w . ^
responses of the residue found in the sample set. ffnecessarv ~
`
samples aid te-analyze to give a responsewithin theso n d a td a ^ 016 ***'
g. Fortifications that bracket the highest residue expected in n t ' sample will be included with each sample set. ff residues T "**
` . outside these limits, additional fortifications wifi be included
. subsequentsampleset to establish thatmethod recoveries are avathhlT * the analyteofinterestat concentrationsexceeding thosem treatedTM * **
Centre Analytical L sbo n o n s. Inc. Snrdy 023-015
f>geI4
' `Centre-Analytical Iaboratories^ac;--
Centre A nalytical Laboratories, Inc. `
`
-Page 24 o f 38
. - Page 85 o f 102
Centre Study N o.: 023-063
" CentreProtocolNo. OOP-023-063
:.
' CmtreMethodHo; oOM^GMli
' . h. Fortification recoveries vrithin 60 to I30S are aceatablt for f,,--
' at the LOQ level. Recoveries between 70 a 120%-
,"
fortifications at levels,greaterthan da LOQ. K hre to
f.
. teqtrires an invcsrigarioa of eaase and a fiiil teanaiysis of fte
'
peaks are detected at toecotresponamganalyteretention times bmare (than theloweststandardmfi bernxatcdasNQ(iwqo^y)|e^
, '
.-
.t
j. Background levels of analyte found in'commlfblank sanmles th
correspond to wines below the LOQ, but are sriB
...T j
used tocone fortificationrecoveries.-
* * '. ^ w)u "e
: j- If samnles are notloadedon extracted, samples mostbe stt nntDanalysisandanalyzedpreferablywithina wee -
are to6*C
Recoveries from metfiod development for all matrices can be foundT
: TablesI-YL * ,
` *
/ .
^
a
t .
'n
4.5. Phhformanch.Cruesu . .
- ' t/.
*
* *
* . **
-
The following two critreia must fie met before the burial analysis of satnpla. " '
esoecialiy. what using 'diffaeat instrumentation set-ups-t&m those cited in this '
method. _
\ . '
. f.
First Criterion - Inject a satxfirdsolmion on theLOMS/MScotresoondine to'the
estimated LOQ (10 ppo LOQ ir equivalent to a standard solution 'of 0.2 ngftnL)
and obtain a signal to noise ratio of at lest 9:1 rthnV to the reagent blank. jf
this criterion cannot be met,- optimize antt change .instrument opcatin
panmeten.
': . " . ' '
' .
:
I
Second Criterion *Meet a ret ofstandards tangjngfroma orhelm
u,
to the highestcoocentrntionJeyeL GenerateacaDbtationcarve fortbcamiw.
' obtain a linear regression with aCoefficient ofdcterminarion(r2) of at least 0.9S5
for the analyte. Once this criterion has been deaonaaed. samples may be
` analyzedwith standards interspetsed.
'-
*.
.
*
Centre AnalyticalLaboratories. lot Swdy*023-015
-CentreAnalytical Laboratories, Iric. Centre A nalytical Laboratories, Inc.
! hje!5
Page 25 o f 38 Page 86 o f 102
Centre Study N o.: 023-063
CentreProtocol No.'00P^)23-063
C tatrt Method No:00M-023-015
4.6. TimeRequiredforAnalysis
.
.
A set of 14 samples can be taken thronjh the extraction
'
approximately three horns by- one person. ' The LC/MS/MS **?cTiure *n
- standardsand i4 samples) win takeapproximately6 hoars. ' . 3113 (8-10
5- .- CALCULATIONS.
5.1 Analyte Found: '
t^icauB ute annum oi auiyie round {a ngrmc, h^ d n standardcarve genented bythe MaeQuansoftwareprogram usin
.Equation 1: -
*
`
,'
'
Analytefound fngfmD = freak area - iwctteafl
..
slope- "
- 3.2 ComponentResidueConcentration `
.
__ "
'
. . Determinethe component residue concentration usingEouhtron 2
Equation!; *
( " UIg ut
' I.
' Residue found (ntt/g) * . ".
(ng/m L )xD FxFV ^nj^
sample weight (g)
~
WhereDF=dilution factorandFV= final volume
22s< ng^=ppb
-
.
' -'
i
5 J PESCSNrRECOVHtY
`
Calculate the percent tccovety for samples fortified with known
analytespriorto extraction, fromEquation 3.
. ounts f
Equation3: .
;. *
(ng/g found- avengeng/gfoundin control! Recovery^) =
! ' ng/gadded [
.
;Car: AmlytieelUontories.Inc.StudySQS-OU
Past 16
CentreAnalytical-Laboratories, Inc.Centre A nalytical Laboratories, Inc.
''
-- - -- Page 26 of3$ Page 87 of. 102
Centre Study N o.: 023-063
Cernie ProtocI"N:'00P^l23-063
Centre Method No: 00M-023-015
- SAFETY'
Thera are no unusual hazards associated with this method. The anal
'
read the material safetydatashees forall reagents before penormm siloul<i
Normal laboratoryprecautions should be talcea.
a method.
' t
Crane Analytical Laoiatcries. h t Stod 023-015
'Centre Analytical'Laboratories,be.'-- Centre A nalytical Laboratories, b e,
P a se l7
Page 27-of38 Page 88 o f 102
Centre Study N o.: 023-063 CentreProtocol No'. 0OP-O23-O63
* Centre Method No: OOM-023-oiS
i
TABLES
Cenre Ait3hQC3lLsba(siories.Iitc.S{udT*(C34}15- *
* .
.
.
*.
. Pajc18
'Centre Analyticallaboratories, Inc. Centre A nalytical Laboratories, Inc.......
: ................... ...
Page 28 b38 ' Page 89 o f 102
Centre Study N o.: 023-063
CentreProtocol Na OOP^023^63
Centre Method No: O O M ^O Ii
Table fc Summary o f Recoveries forPFOS in Quail Egg Membrane
s flc . ; `
F o ille v d "
(pob)
SSea^v
'
0001791 MamaBlank A
0003791MariaBlank B .
000379 ManiaBlank C
0003791Spk A
. 0003791SpfcB
. 000379tSpkC
0 0
:o
10
w
- 10
. NA
NA
- A '
' 95
92
' 000379SpkD 0003791SpkE 000379SpkF
.
' SO
so
. 92
* 95 96 '
000379SpkG
.
250
. 93 .
000379SpkH
. 000379SexI '
'
250
250 .
95 93 '
. AVERAGE: * J3
STANDARD DEVIATION
i.
. ; : RELATIVESTANDARDDEVIATION: _
3
;
* ' '
.
'
T ab le IL- Sum m ary o f R ecoveries fo r PF O S in M allard E K Membrane '
S&OB6 . "
F a tlrv d
feoM
SR eesw y
'
0003E M ar BlankA 00Q38C2MasixBlank B
0 NA
* 0 . - "NA '
00038Q2M assBlank C * . 0 ' na' `
ooaamspkA.
10
106 '
0003302SpkB
10
: 122 * :
000382 SpkC
'
10
106 ' -
000388SpkD.. - OOBS&SpirH
..
30 -
` ` 96
- 96 ' *
WBSSSpkF .
sa . .
90 .
OOtBJtfiSpkG .
2
. 93 .
'
ootBaaipfcH
250
.. ' 92 ' :
000382SokI *.
- *
25 .
` AVERAGE:
93
99 '
' ~
..
STANDARDDEVIATION:
10 .
. RELATIVE STANDARD DEVIATION:
10 .
Na = NotApplicable
Centre Analytical Lataanries, Ine. Study #023415
Cffltre'Analytical'laboratories, Inc.-' Centre A nalyticai Laboratories, Inc.
ftp: 19
. Page 29 o f 38 Page 90 o f 102
Centre Study N o.: 023-063
C ^S nsraocl-N a 00P:g23-063
Centre Method No: 0 0 M - q 2 3 - 0 I S
TableIQ; Summary orRecoveries for PFOS in Quad E Yolk
Srapte
0WSMaanBlankA C795 MarieBUakB 3S ManiaBhnle.C
OraSSjk
: 0003795SpkB 0003795SRkC 0003795SpkD 1 0003795 SpkE
. 0003795SpkF 0003795SpkG
0003795SpkH .
Fort. Lrrd
fwfe) 0 0 0 . 10.
10 . 10
50 50 50 250
250
` `*2eoie^
~T
HA HA.
a a.
70 ' 72 ' 10 13 IS
_00Q3795SpkT
250 *
AVERAGE: .STANDARD DEVIATION: RELATIVESTANDARDDEVIATION:
g
71
5.
11
TableIY: Summary ofRecoveries forPFOS-ln Mallard Eg
NA = NotApplicable '
AVERAGE: ' STANDARDDEVIATION: RELATIVESTANDARDDEVIATION:
Ceare Aralytial Laboaoria. 1rs. Sadr 023-015
CentreAnalyticallaboratories, Inc----Centre A nalytical Laboratories, Inc.
hie20
-' Page30of38 Page 91 o f 102
Centre Study N o.: 023-063
'Centre'Protocol No. OOP-023-063
Centre Method No: 00M-023-015
Table V: Summary ofRecoveries for PFOS in Quail Egg Albumen
. S apte *
* * FortLevd
SR CSHBj .
(W*>)
0003296 Manx Bhnk A 3796MatrixB hnk3
' 000796 U nix S la b C
.
0. 0 . o'
r
* .
0003796SpiA '
w *.
003796SpkB
10. -
'
0003796 SpkC
.
10.
-
003796SpkD
:
0003796SpkE 0003796SpkF
. 0003796 SfkO . 0003796SpkH
'.
.' - JO '
2
. . 250 . .
*'
0003796SokI
250 ' ' . AVERAGE:
_. t<
. STANDARDDEVIATION: * RELATIVESTANDARDDEVIATION:
.
Ha Na NA . 79 93 X 96 95 90 97 102 102 54 7* 7
-- *
.
'. .
"" '
'
TableVI: Summary ofRecoveriesforFOSin Mallard Eg* Albumen ' '
** Satwpl
.* Eon.Lewi
.
ftilecsvtty
. .(nokV
0003100Manx Bbak A 0003100Manx BbakB 0003.100M anx Blank C
0003100SpkA.
' .
. .
. o.;.
0. 10
'
. . Na
..
na
' na
.. as
" *
** * .
V .<
. 0003100SpkB OOOHOOSpkC '
. .
10 : 10 .
ts .B
'.
! *
. .0003100SpkD OOOSSOOSpkE
. 30 ... - -I * '
96 ' . I0 * *
* **
0603100Spk? 'OOQ3SOOSpkG` ' '
50. ' 250 * '
. .-
100 . 98 * .
)
. 0003100SpkH '
250 :
102 *
` '*
-- __ ^.0003100 SokI ' '
2 . _ 101
,
. . ; AVERAGE: ' STANDARDDEVIATION: '
96 7
`
., RELATIVESTANDARDDEVIATIOtfe
7`
. . NAs NotApplicable
CiamAsaljdtaloeaoric^IntS^iQ^^U .
-entre-Analyticallaboratories.Inc. - - - - -------; . .---- -------------- Eage.,3.1 of 38 ' . .
Centre A nalytical Laboratories, Inc,
` Page 92 o f 102
Centre Study N o.: 023-063 fo.'00P^023^063'
Centre Method No: OONfor^j-
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CrtreHotocol No:0dpt(3233363-
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Page 35of 38
Page 96 o f 102
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................^ Cratre ProtocolNo. 0OP-O23-Q63
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Page 37 of 38 Page 98 o f 102
Centre Study N o.: 023-063
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Page 38 of 38 Page 99 o f 102
Centre Study No.: 023-063
Centre Analytical Laboratories, Inc.
3048 R esearch D riv e, S tate C ollege, PA 16801.
Phone: (814)231.8032, Facsim ile: (814)231-1253
PROTOCOL AMENDMENT Amendment Number I Effective Date: 3/23/01
Page 1 of 1
Centre Study Number 023-063
Centre Protocol Number 00P-023-063
DESCRIPTION OF AMENDED SECTION
1. Title 2. 8Analytical Method 4.4.4.f.
AMENDED TO
1. Change the analytical phase title to read: Extraction of Potassium Perfluorooctanesulfonate from Egg Membrane, Albumen, and Yolk for Analysis Using HPLC-Electrospray/Mass Spectrometry (This will also change on Page 2 and Page 3)
2. Change the first sentence to read: Fortification recoveries within 70-120% are acceptable for fortifications at the LOQ level.
RATIONALE
1. Per Sponsor request to maintain consistency with method. 2. Reflect the requirements that were followed during the method validation.
IMPACT ON THE STUDY
1-2. No negative impact on the study.
ft P^kJlvv
Study Director SCiigmniQattuiirre
Date
'm o j
Date
Centre QAU Review AYW
February 12. 1998/1
Centre A nalytical Laboratories, Inc.........................................
Page 100 o f 102
Centre Study No.: 023-063
Centre Analytical Laboratories, Inc.
3048 Research Drive, State College. PA 16801.
Phone: f814)231-8Q32. Facsim ile (814)231-1253
PROTOCOL AMENDMENT Amendment Number 2 Effective Date: 3/26/01
Page 1 of 1
Centre Study Number. 023-063
Centre Protocol Number 00P-Q23-Q63
DESCRIPTION OF AMENDED SECTION
1. Section 9: Analytical Method 2. Section 9: Analytical Method 3. Section 10: Experimental Design
AMENDED TO
1. Amend Page 21 Section 4.3.2 a. to read: Prior to weighing and extracting the egg membrane samples, the membranes will be rinsed with ASTM Type IH20 and patted dry.
2. On Page 24 Section 4.4.4.d, remove: One ASTMTypeTwater blank
3. Amend Page 6 last paragraph, second sentence to read: For egg yolk and albumen, ~ 0.5 g samples and egg membrane - 0.1 g samples will be
weighed and dried in an oven at - 105C for.- 12-14 hours.
RATIONALE
1. At the instruction of the study director to ensure that no contamination of egg yolk or albumen is present in the sample.
2. This sample is only to check for PFOS residue present in the ASTM Type IH 20 used. 3. To account for sample availability.
IMPACT ON THE STUDY
1-3. No negative impact on the study.
A u , PStudy Diiinrecrtnor SSiiogmnaattuiir*e
U
U ill Date
Date/ ' Centre QAU Review %
//A
February 12, 1998/1
Centre A nalytical Laboratories, Inc.
Page 101 o f 102
Centre Study No.: 023-063
3048Research Drive,State College, PA 16801.
Phone: (814)231-8032,Facsimile: (814)231-1253
PROTOCOL DEVIATION
DeviationNumber 1
Date ofOccurrence: (1)03/27/01,(2)04/02/01
Centre Study Number 023-063
Centre Protocol Number OOP-023-063
DESCRIPTION OF DEVIATION
1. 8Analytical Method 4.4.4.f: Quantified Centresample 0103196 and its duplicate injection when the concentrations were greater than that of the highest standard in Set 032601C
2. Amendment #1: Accepted recoveries of58% and 61%for Centre sample 0003800 Spk A and its duplicate injection in Set 033001C.
ACTIONS TAKEN i.e.. amendment issufflLSQP.wviaon. etc.,,
1-2. Protocol deviation issued.
Recorded By/Date:
IMPACT ON THE STUDY
1. No negative impact because the areas were < 5%greater than the highest standard.
2. No negative impact because Spk Bgave acceptable recoveries and sample results reported in the set were onlyfor confirmation.
S $ 1 )W L U _
Study Diirreeccttnorr SSigonnatnuire --
v
Date
[o 1 Date
/J 9 //
Date
q/(>/ol
CALQAU Review -K&. i f a l t l
February 12. 1998/2
Centre A nalytical Laboratories, Inc.
Page 102 of 102
