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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
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Centre Study No.: 023-063
GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT
Centre Study Number 023-063, entitled "Extraction o f Potassium Perfluorooetajaesulfonate from Egg Membrane, Albumen, and Y olk for A nalysis U sing HPLC-Electroapray/Maas Spectrometry," conducted for 3M Environmental Technology and Safety Services, was performed in com pliance with US EPA TSC A G ood Laboratory Practice Standarda (40 CFR Part 792) by Centre Analytical Laboratoriea, Inc with the follow ing exceptions. 1. 8792.63. M aintenance and Calibration o f Equipm ent N o equipment SOP was in
place for the bead beater used in the extraction at the tim e o f use.
Principal avestig a io r Centro Analytical Laboratoriea, Inc.
Saan vrauagnar Study Director W ildlife International, Ltd.
Centre Analytical Laboratoriai, In.
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Centre Study No.: 023-063
QUALITY ASSURANCE STATEMENT
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
1. Protocol Review
2. Extraction, Fortification
3. Raw Data Review
4. Raw Data and Draft Report Review
5. Final Report Review
Date Inspected
Date Reported to Date Reported to
Centre
Study Director and
Management Sponsor Management
3/30/01
4/9/01
5/7/01
3/30/01
4/9/01
5/7/01
4/5-9/01 4/17-18/01
4/18/01 4/26/01
5/7/01 5/7/01
6/28/01
9/6/01
9/6/01
Ange a Morgan Qualit Assurant
ditor
% Date
Centre Analytical Laboratories, Inc.
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cnoii
C a t Study No.; 023-063
CERI TON Q F A IM E M C T T
This report, for Centre Study Number 0234)63, it a true and com plete representation of the a w data tot the Study-
Submitted by.
C ent Analytical Laboratories, Inc. 3048 Research Drive State College, PA 16801 <814)231-8032
Principal Investigator, Centre;
ly ^Decker
Scientist Centre Analytical Laboratories, Inc.
Centre Analytical Laboratories, Inc, Facility Management:
/ f a ' t y t o d ' 14
/ Richard A. ^ n z 2 in V m P ( President Centre Analytical Laboratories, Inc.
__________
Study D irector
D a
D a <5^, f-uev-oi
Sean Gallagher W ildlife International, L td
Sponsor Representative
*
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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: OOP-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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PROJECT PERSONNEL
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 lin g 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 PA G E ............................................................................................................................ 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 n. 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 B lanks...................................................... 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 VIE. Summary of PFOS for Week 1 Mallard Membrane Control Samples............26
Table IX. Summary of PFOS for Week I 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 XU. Summary of PFOS for Week 1 Mallard Albumen Samples at 6.2 ppm a i .... 28
Table XIII. 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 o f PFOS for W eek 1 M allard Y olk Sam ples at 17.6 ppm a i......... 30
Table XVQ. 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 XXII. 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 B lanks.............................................35
Table XXVUI. 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 Sam ples..........38
Table XXXIQ. 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 Sam ples............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 XXXVDI. 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 a i............. 42
Table XLI. Summary of PFOS for Week 6Quail Membrane Control Samples................42 Table XLH. Summary of PFOS for Week 6Quail Membrane Samples at 6.2 ppm a i..... 43
Table XT.ITT. Summary of PFOS for Week 6Quail Membrane Samples at 17.6 ppm a i.. 43
Table XLIV. Summary of PFOS for Week 6Quail Albumen Control Samples...............44
Table XLV. Summary of PFOS for Week 6Quail 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 XLV1I. Summary of PFOS for Week 6Quail Yolk Control Samples.................... 45
Table XLVUI. Summary of PFOS for Week 6Quail Yolk Samples at 6.2 ppm a i ..........46
Table IL. 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 UHL Summary of Moisture Determinations for Quail Membrane Sam ples......... 50 Table LTV. Summary of Moisture Determinations for Quail Albumen Samples............ 51 Table LV. Summary of Moisture Determinations for Quail Yolk Samples....................52
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Figure 1.
LIST OF FIGURES Page
Typical Calibration Curve for PFOS.................................................................54
Figure 2. Chromatogram Representing a 0.1 ng/mL standard for PF O S......................55
Figure 3. Chromatogram Representing a 1.0 ng/mL standard for PF O S......................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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0002.
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1.0 SUM 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 OBJECTIVE
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 INTRODUCTION
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 ED
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 R E FE R E N C E M A T E R IA L
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
0
1c8f17s -- 0-
= =
Perfluorooctanesulfonate 499 (CgFnSCV)
Note: The neutral molecule and standard from which the PFOS (anion) is
obtained, is potassium perfluorooctanesulfonate [C8F 17SO3K], molecular weight
538.
6.0 DESCRIPTION OF ANALYTICAL M ETHOD
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.
E gg 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 o f 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 w ere less than the low est 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 o f Instrument and Operating Conditions Instrument: PE SCIEX API 3000 Biomolecular Mass Analyzer
Interface: SCIEX Turboion Spray Liquid Introduction Interface
Computer: Dell UltraScan P I 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, 4p 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 Transition Monitored
negative
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
Centre Analytical Laboratories, Ine.
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
6.6 Quantitation and Example Calculation
Set
12
14 4 350C
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 = D ilution 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 quanitifiable 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
ss From equation 2:
Analyte found (ng/g) =
0.36 ng/mL
(0.36 ng/mL x 25 mL x 11 1.0038 (g)
From equation 3: % Recovery
= 8.97 ng/mL
= (8.97 ne/g x - 0 ng/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 DESIG 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 H 2O
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-XXXI. 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 nonquantifiable levels to 65,500 ng/g in the quail yolk samples. These results are reported on a w et w eight basis. Individual results are listed in T ables X X X II-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 CONCLUSIONS
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 RETENTION OF DATA AND SAM 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 03290IB 03290IB 033001A 033001A 033001B 033001B 033001C 033001C
Extraction Analysis Date Date
3/26/01 3/26-27/01
3/26/01 3/26-27/01
3/26/01
3/27/01
3/26/01
3/27/01
3/26/01
3/27/01
3/26/01
3/27/01
3/27/01
3/28/01
3/27/01
3/28/01
3/27/01 3/28-29/01
3/27/01 3/28-29/01
3/27/01
3/29/01
3/27/01
3/29/01
3/28/01
3/30/01
3/28/01
3/30/01
3/28/01
3/30/01
3/28/01
3/30/01
3/29/01
3/31/01
3/29/01
3/31/01
3/29/01 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 (nob)
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 Analytical Laboratories, Inc.
Page 23 o f 102
Centre Study No.: 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 NO 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 3/26-27/01
3/26/01 3/26-27/01
3/27/01 3/28-29/01
3/27/01 3/28-29/01
3/30/01
4/2/01
3/30/01
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 Analytical Laboratories, Inc.
Page 24 o f 102
Centre Study No.: 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 Spk B* 0003802 Spk A 0003802 Spk A* 0003802 SpkB 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 3/26/01 3/26/01 3/27/01 3/27/01
3/27/01 3/27/01 3/27/01 3/27/01 3/29/01 3/29/01
10 10 100 100 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 SpkB 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 032701B 03270IB 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 Analytical Laboratories, Ine.
Page 25 o f 102
Centre Study No.: 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 Spk B* 0003799 Spk A 0003799 Spk A* 0003799 Spk B 0003799 Spk B*
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
3/27/01 3/27/01 3/27/01
10 10 98
3/26/01 3/27/01
3/27/01 3/28/01
98 10
3/27/01 3/28/01
10
3/27/01 3/28/01
97
3/27/01
3/28/01
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 E01-0263-20852* E01-0263-20853 E01-0263-20853* E01-Q263-20854 E01-0263-20854*
EOI-0263-20842
EOI-0263-20842* E01-0263-20843 EOI-0263-20843*
C en tre
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
Analyte
Date Date Found (ppb)
3/26/01 3/26/01
3/27/01 3/27/01
NQ NQ
3/26/01
3/27/01
NQ
3/26/01
3/27/01
NQ
3/26/01
3/26/01
3/26/01
3/27/01
3/27/01
3/27/01
NQ
NQ
NQ
3/26/01
3/27/01
NQ
3/26/01 3/26/01
3/27/01
NQ
3/27/01 ______ NQ____ AVERAGE: NQ
STANDARD DEVIATION: NQ
NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/xnL)) LOQ = 10 ppb
Centre Analytical Laboratories, Inc.
Page 26 of 102
Centre Study No.: 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 EOI-0263-20846* E01-0263-20832 E01-0263-20832* E01-0263-20847 E01-0263-20847* 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 (nob)
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 E01-0263-20850* E01-0263-20857
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
* Duplicate Injection
LOQ = 10 ppb
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 3/26/01
3/27/01 3/27/01
3/26/01
3/26/01
3/27/01
3/27/01
3/26/01 3/26/01
3/27/01 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
** 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 27 of 102
Centre Study No.: 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* E01-0263-20842 EOI-0263-20842* EOl-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 3/26/01 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
3/26/01
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
EOI-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
3/26-27/01
3/26/01
3/26-27/01
3/26/01
3/26-27/01
3/26/01
3/26-27/01
3/26/01
3/26/01
3/26-27/01
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)
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/niL)) LOQ = 10 ppb
Centre Analytical Laboratories, Inc.
Page 28 o f 102 coo
Centre Study No.: 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 EOI-0263-20856* EOI-0263-20856 EOl-0263-20856* EOI-0263-20850 EOl-0263-20850* E01-0263-20857 E01-0263-20857* E01-0263-20851 EOI-0263-20851* EOl-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 Date
Analysis Date
3/26/01
3/27-28/01
3/26/01
3/27-28/01
3/30/01
4/2/01
3/30/01
4/2/01
3/30/01
4/2/01
3/30/01
4/2/01
3/26/01
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)
3550** 3490** 1388** 1369** 845** 771**
24.7 23.9 23.8 23.3 78.0** 69.7** 19.7 19.8 22.5 2JO
Table XIV. Summary of PFOS for Week 1 Mallard Yolk Control Samples
Sponsor ID
E01-0263-20852 E01-0263-20852*
E01-0263-20853
E01-0263-20853* E01-0263-20854 E01-0263-20854* E01-0263-20842 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
032601B
032601B 032601B 03260IB 032601B 032601B 03260IB 032601B
Extraction
Analysis
Analyte
Date Date Found (ppb)
3/26/01
3/26/01
3/26/01
3/27/01
3/27/01
3/27/01
NQ
NQ
NQ
3/26/01 3/26/01 3/26/01
3/27/01 3/27/01 3/27/01
NQ NQ NQ
3/26/01
3/27/01
NQ
3/26/01 3/26/01
3/27/01 3/27/01
NQ NQ
3/26/01
3/27/01 ...........n q AVERAGE: NQ
STANDARD DEVIATION: NQ
NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) LOQ = lOppb
** 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 No.: 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 E01-0263-20846* E01-0263-20832 E01-0263-20832* E01-0263-20847 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 032601D 03260ID 032601D 032601D 032601D 032601D 032601D 032601D
Extraction
Analysis
Date Date
3/26/01 3/26/01 3/26/01 3/26/01 3/26/01 3/26/01
3/27-28/01 3/27-28/01 3/27-28/01 3/27-28/01 3/27-28/01 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 E01-0263-20856* EOI-0263-20850 EOl-0263-20850* E01-0263-20857 E01-0263-20857*
E01-0263-20851
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 3/26/01 3/26/01 3/26/01 3/26/01 3/26/01
3/26/01
3/27-28/01 3/27-28/01 3/27-28/01 3/27-28/01 3/27-28/01 3/27-28/01
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 Analytical Laboratories, Inc.
Page 30 o f 102
COO f-J
Centre Study No.: 023*063
Table XVII. Summary of PFOS for Week 6 Mallard Membrane Control Samples
Sponsor ID
E01-0263-20993 EOI-0263-20993* EOI-0263-20994 E01-0263-20994* E01-0263-20995 EOI-0263-20995* EOl-0263-20975 E01-0263-20975* E01-0263-20997 E01-0263-20997*
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
Date Date
3/27/01 3/27/01 3/27/01 3/27/01
3/29/01 3/29/01 3/29/01 3/29/01
3/27/01 3/27/01
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(nob)
NQ NQ NQ NQ NQ NQ NQ NQ NQ NO NQ 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* E01-0263-20989 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 3/27/01 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
3/27/01
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 No.: 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* E01-0263-21002 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
3/29/01 3/29/01 3/29/01 3/29/01 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)
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* EOI-0263-20995 EOl-0263-20995*
EOI-0263-20975
EOl-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 03270IB 032701B
032701B
03270IB 032701B 032701B
Extraction Analysis
Date Date
3/27/01 3/28-29/01
3/27/01 3/28-29/01
3/27/01 3/27/01
3/28-29/01 3/28-29/01
3/27/01
3/28-29/01
3/27/01
3/27/01
3/28-29/01
3/28-29/01
3/27/01
3/28-29/01
3/27/01 3/27/01
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 Analytical Laboratories, Inc.
Page 32 of 102
Centre Study No.: 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* EO1-0263-21001 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 03270IB 03270IB 032701B 032701B 03270IB 03270IB 032701B 03270IB
Extraction Analysis
Analyte
Date Date Found (ppb)
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
5.83 6.23 7.92 8.04
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:
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-Q263-20991 E01-0263-20991* E01-0263-20990 E01-0263-20990* E01-0263-21002 E01-0263-21002*
EOI-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
032701B 03270IB 03270IB 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 3/27/01
3/28-29/01 3/28-29/01
3/27/01
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)
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 Analytical Laboratories, Inc.
Page 33 o f 102
Centre Study No.: 023-063
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
Date Date
3/27/01
3/28/01
3/27/01 3/27/01 3/27/01
3/28/01 3/28/01 3/28/01
3/27/01
3/28/01
3/27/01
3/28/01
3/27/01
3/28/01
3/27/01
3/28/01
3/27/01
3/28/01
3/27/01
3/28/01
AVERAGE:
STANDARD DEVIATION:
Analyte Found (ppb)
NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ 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* EOI-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
Analyte
Date Date Found(ppb)
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:
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 Analytical Laboratories, Inc.
Page 34 of 102
Centre Study No.: 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 033001BR
3/29/01 3/30/01
3/31/01 4/2/01
033001BR 3/30/01
4/ 2/01
AVERAGE: STANDARD DEVIATION:
Analyte Found (ppb)
NQ NQ NQ NO 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 3/28/01 3/30/01 3/30/01
3/30/01 3/30/01 4/1/01 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 Analytical Laboratories, Inc.
Page 35 o f 102
Centre Study No.: 023-063
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 03290IB 032901B
Extraction Analysis
Date Date
3/28/01 3/28/01 3/29/01 3/29/01
3/30/01 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 3/29/01 3/29/01 3/30/01
3/31/01 3/31/01 3/31/01 3/31/01 4/2/01
10 10 986 986 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 Analytical Laboratories, Inc.
Page 36 o f 102 00Q & 3G
Centre Study No.: 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
032801B 032801B 032801B 032801B 033001A 033001A 033001A 033001A
Extraction Analysis
Amt
Date Date Added (ppb)
3/28/01 3/28/01 3/28/01 3/28/01
3/30/01 3/30/01 3/30/01 3/30/01
10 10 9963 9963
3/30/01
4/1/01
10
3/30/01 3/30/01 3/30/01
4/1/01 4/1/01 4/1/01
10 99 99
AVERAGE: STANDARD DEVIATION: RELATIVE STANDARD DEVIATION:
% Recovery
82 79 80 83 82 88 93 91 85 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 03290IB 032901B 03290ID 032901D
Extraction Analysis
Amt
Date Date Added (ppb)
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:
%
Recovery
102 104 98 101 87 87 94 93 96 7 7
* Duplicate Injection
LOQ = 10 ppb
Centre Analytical Laboratories, Inc.
Page 37 o f 102
Centre Study No.: 023-063
Table XXXII. Summary of PFOS for Week 1 Quail Membrane Control
Samples
Sponsor
Centre
Set
Extraction Analysis
Analyte
ID
ID
Number
Date
Date Found (ppb)
EOI-0259-20506 E01-0259-20506* E01-0259-20507 E01-0259-20507* EOI-0259-20520 EOl-0259-20520* E01-0259-20521
0101987 0101987 0101991 0101991 0102043 0102043 0102047
032901A 032901A 032901A 032901A 032901A 032901A 032901A
3/29/01 3/29/01 3/29/01 3/29/01 3/29/01 3/29/01 3/29/01
3/31/01 3/31/01 3/31/01 3/31/01 3/31/01 3/31/01 3/31/01
5.20** 5.57**
NQ NQ NQ NQ NQ
E01-0259-20521* E01-0259-20509 E01-0259-20509*
0102047 0101999 0101999
032901A 032901A 032901A
3/29/01
3/31/01
3/29/01
3/31/01
3/29/01
3/31/01
AVERAGE:
STANDARD DEVIATION:
NQ NQ NQ NQ NQ
Table XXXIII. Summary of PFOS for Week I 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 E01-0259-20527* E01-0259-20515 E01-0259-20515* E01-0259-20528 E01-0259-20528*
0102067 0102067 0102019 0102019 0102071 0102071 0102023 0102023 0102075 0102075
032901D 032901D 032901A 032901A 032901A 032901A 032901A 032901A 032901D 032901D
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
4/1-2/01 4/1-2/01 3/31/01 3/31/01 3/31/01 3/31/01 3/31/01 3/31/01 4/1-2/01 4/1-2/01 AVERAGE:
18800** 17900**
127 129 123 116 172 172 11500** 11500** 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 vaiues 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 Analytical Laboratories, Inc.
Page 38 o f 102
g c q s //<
Centre Study No.: 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 EOI-0259-20532* EOl-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 03290ID 032901D 032901D 03290ID 032901D 032901D 03290ID
Extraction Analysis
Date Date
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
4/1-2/01 4/1-2/01 4/1-2/01 4/1-2/01 4/1-2/01 4/1-2/01 4/1-2/01 4/1-2/01 4/1-2/01 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*
EOl-0259-20521
EOI-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 03280IB 032801B
03280IB
032801B 03280IB 03280IB
Extraction Analysis
Analyte
Date Date Found (ppb)
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
3/30/01 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
NQ NQ NQ NQ NQ NQ
NQ
NQ NQ NQ
AVERAGE: STANDARD DEVIATION:
NQ NQ
* Duplicate Injection
NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) LOQ = 10 ppb
Centre Analytical Laboratories. Inc.
Page 39 o f 102
C O O ' sc*
Centre Study No.: 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* E01-0259-20527 E01-0259-20527* 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 03280IB 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 EOI-0259-20529* EOl-0259-20531 EOI-0259-20531* EOl-0259-20532 E01-0259-20532* E01-0259-20518 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
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:
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 Analytical Laboratories, Inc.
Page 40 o f 102
Centre Study No.: 023-063
Table XXXVIII. Summary of PFOS for Week I Quail Yolk Control Samples
Sponsor
ID
E0I-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
Date Date
3/28/01 3/28/01 3/28/01 3/28/01
3/30/01 3/30/01 3/30/01 3/30/01
3/28/01 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 3/30/01
AVERAGE:
STANDARD DEVIATION:
Analyte
Found (ppb)
NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ
Table XXXIX. Summary of PFOS for Week 1 Quail Yolk Samples at 6.2 ppm al
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 EOI-0259-20528*
Centre ID
0102065 0102065 0102017 0102017 0102069 0102069 0102021 0102021 0102073 0102073
* Duplicate Injection
Set Number
032801 AD 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
3/31/01 3/31/01 3/31/01 3/31/01 3/31/01 3/31/01 3/31/01
3/28/01 3/28/01 3/28/01
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 Analytical Laboratories, Inc.
Page 41 of 102 G 0 0 ? ,3
ssssr-sgp
Centre Study No.: 023-063
Table XL. Summary of PFOS for Week 1 Quail Yolk Samples at 17.6 ppm ai
Sponsor ID
Centre ID
Set Number
Extraction Date
Analysis Date
Analyte Found(ppb)
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*
0102077 0102077 0102085 0102085 0102089 0102089 0102033 0102033 0102093 0102093
032801AD 032801AD 032801AD 032801AD 032801AD 032801AD 032801 AD 032801AD 032801 AD 032801 AD
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(ppb)
E01-0259-20735 E01-0259-20735* E01-0259-20736 E01-0259-20736* EOI-0259-20737 EOl-0259-20737* E01-0259-20720 E01-0259-20720* E01-0259-20738 E01-0259-20738*
0102903 0102903 0102907 0102907 0102911 0102911 0102843 0102843 0102915 0102915
* Duplicate Injection
033001B 033001B 033001B 033001B 033001B 033001B 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
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:
NQ NQ 9.85** 8.93** 15.6** 14.4** 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 Analytical Laboratories, Inc.
Page 42 of 102
Centre Study No.: 023-063
Table XLII. Summary of PFOS for Week 6 Quail Membrane Samples at 6.2 ppm ai
ID
E01-Q259-20742 EOI-0259-20742* E01-0259-20743 E01-0259-20743* E01-0259-20744 E01-0259-20744* E01-0259-20745 EOI-0259-20745* EOl-0259-20746 E01-0259-20746*
ID
0102931 0102931 0102935 0102935 0102939 0102939 0102943 0102943 0102947 0102947
Number
03300IB 033001B 033001B 033001B 033001B 033001B 033001B 033001B 03300IB 033001B
Date Date
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
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
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
EOI-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
03300IBD 033001BD 033001BD 033001BD 033001BD 033001BD 033001B
03300IB
033001B 033001B
Extraction Analysis
Date Date
3/3a m 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
3/30/01 3/30/01 3/30/01 3/30/01
4/2/01 4/2/01 4/2/01 4/2/01
AVERAGE:
STANDARD DEVIATION:
Analyte Found (ppb)
405 389 318 318 394 400 262 250 186 185 311 86.6
Centre Analytical Laboratories, Inc.
Page 43 of 102
GOO .*v.
Centre Study No.: 023-063
Table XLIV. Summary of PFOS for Week 6 Quail Albumen Control Samples
Sponsor ID
EOI-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
4/1/01 4/1/01
3/30/01
4/1/01
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
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
4/1/01 4/1/01
3/30/01
4/1/01
3/30/01 3/30/01
4/1/01 4/1/01
3/30/01
3/30/01
4/1/01
4/1/01
3/30/01 3/30/01 3/30/01
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
Centre Analytical Laboratories, Inc.
Page 44 of 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)
EOI-0259-20747 E01-Q259-20747* 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
Centre
Set
Extraction Analysis
Analyte
ID ID Number Date Date Found(ppb)
E01-0259-20735 E01-0259-20735*
0102901 0102901
032901B 032901B
3/29/01 3/29/01
3/31-4/1/01 3/31-4/1/01
NQ NQ
E01-0259-20736 E01-0259-20736* E01-0259-20737
0102905 0102905 0102909
032901B 032901B 032901B
3/29/01 3/29/01 3/29/01
3/31-4/1/01 3/31-4/1/01 3/31-4/1/01
NQ NQ 51.5**
E01-0259-20737*
0102909
032901B
3/29/01 3/31-4/1/01
51.1**
E01-0259-20720
0102841
032901B
3/29/01 3/31-4/1/01
NQ
E01-0259-20720* E01-0259-20738
0102841 0102913
03290IB 03290IB
3/29/01 3/29/01
3/31-4/1/01 3/31-4/1/01
NQ ND
E01-0259-20738*
0102913
03290IB
3/29/01
3/31-4/1/01 AVERAGE:
ND NQ
* Duplicate Infection
STANDARD DEVIATION: 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 Analytical Laboratories, Inc.
Page 45 of 102 0 0 0 2 ,8 9
Centre Study No.: 023-063
Table XLVIII. Summary of PFOS for Week 6 Quail Yolk 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* EOI-0259-20745 EOl-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 03290ID 03290ID 03290ID 03290ID 032901D 03290ID 032901D
Extraction Date
Analysis 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^ 36100^ 23600. 23600" 40000\ 40600" 24600v 25800^ 33100 7610
Table IL. Summary of PFOS for Week 6 Quail Yolk Samples at 17.6 ppm ai
Sponsor ID
E01-0259-20747 E01-0259-20747* E01-0259-20748 E01-0259-20748* E01-0259-20749 EOI-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
03290ID 032901D 03290ID
Extraction Analysis
Date Date
3/29/01
4/1-2/01
3/29/01 3/29/01 3/29/01 3/29/01
4/1-2/01 4/1-2/01 4/1-2/01 4/1-2/01
3/29/01
3/29/01
4/1-2/01
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 465(Xr
59400N
59300^ 56800 57800 56500 6510
NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL))
Centre Analytical Laboratories, Inc.
Page 46 o f 102 e sc o n o
Centre Study No.: 023-063
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 EO1-0263-20832 EO1-0263-20847 E01-0263-20848 E01-0263-20856 E01-0263-20850 EO1-0263-20857 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 EO1-0263-21003 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
Centre Analytical Laboratories, Inc.
Page 47 o f 102
Centre Study No.: 023-063
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
EOl-0263-20991
EO1-0263-20990 E01-0263-21002 E01-0263-21003 EOl-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
Centre Analytical Laboratories, Inc.
Page 48 o f 102
Centre Study No.: 023-063
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 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 EOl-0263-20971 E01-0263-20989
EO1-0263-20991
EOl-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
% M oisture
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
Centre Analytical Laboratories, Inc.
Page 49 o f 102
Centre Study No.: 023-063
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 % Moisture
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 Analytical Laboratories, Inc.
Page 50 of 102
Centre Study No.: 023-063
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
E01-0259-20747
E01-0259-20748 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
% M oisture
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
Centre Analytical Laboratories, Inc.
Page 51 of 102
Centre Study No.: 023-063
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
EOl-0259-20747
E01-0259-20748 EOl-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
% M oisture
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
Centre Analytical Laboratories, Inc.
Page 52 o f 102
Centre Study No.: 023-063
FIGURES
Centre Analytical Laboratories, Inc.
Page 53 o f 102
Centre Study No.: 023-063
Figure 1. Typical Calibration Curve for PFOS
Centre Study No.: 023-063
Figure 2. Chromatogram Representing a 0.1 ng/mL standard for PFOS
Sm * u r y p i i
AC*. MuiC o n cen tratio n
Acq. Tin#
/t . l M
01 21/01 02i 5s20 PM
M odified*
ta w U a i factor i
UVUnomale.#mftulmWmiMMetiledldtl
RT Window B o a c e i d RTi o o . NlCCAi o p . R oi*ti b p . PMk Ratio Kao. A4). R atlo t
R*p. i l . Ratio Oao R e la tiv e Rft
T15o.0o
21.30 2
30.0 4.43 0.20
50..0010
4.00
3.00
l o t . typo
aroaR o to o tio a T la ti
H olftlti
t a r t T tM i Rod TUm i
4.44 i
}.1S001o*.302. 4.34 i 4.50 i
Centre Analytical Laboratories, Inc.
Page 55 of 102
OOG 399
Centre Study No.: 023-063
Figure 3. Chromatogram Representing a 1.0 ng/mL standard for PFOS
SIMM*V1li TInydpoxis
AC. O ltC i T lM i
lu a ttrd 1.060
63/36/01 03i)7tl7 nt
n o d lflo d i N M U if rrn o ti
MAxl*MflTvlUNMMhodldii
XT windows b p e t 4 KTt ~ p . W idth. Sp. H olfhti ' P . *M * lUtlOl
p . M l. ftotioi
<iup . Mv *iel.tivlu*cmier..
3S9..3006
a
10.0 I . 4J
0.20 0.01 3.00
4J..0 0
OO
ta t. typi U tM C iM f ia it CM .
M alpbti l u r e Tlaat Cad T lM s
k l ! To Shoo 4 .4 4 mil
`>343.4 cm ICO 1.13003 epo
4.39 a la 4.41 a la
Centre Analytical Laboratories, Inc.
Page 56 of 102 GV.< Ot n ; v '*ui A\ >
Centre Study No.: 023-063
Figure 4. Chromatogram Representing a Reagent Blank for PFOS (Centre ID Reagent Blank A, Set: 032601A)
la m it la d n i v v l t typsi C eK M tm im i A e a . du i
A. TfcMi
IS
B la ah s .s s o 03/31/01
0 4 i U i )< fM
M odiClad<
Taa
aaeM lao F a c ta r:
1
M ela* T hraah o ld i
t.U
a t m Tliroafcoldi
23.30
NUB. f e N t k l l
2
RT Wladowi
30.0 aaa
lip a c c a d Rfi
4.43 la
S ap. M idtiii
0.20
la p . Natalie i Sap. Fat* R atio i
0.01 3.00
1
Bap. A dj. R aclai Bap. v a l. R acial Oao R a l a t l v a NTt
4.00 3.00 MO
1 !
X at. Typai R acaotiaa t u a i A caai
Naiptifci
cart tlaai Bad T laa i
M aul
MMa.a4.43 la
*H .3 I . U h W I CB8
4.37 4.31 a l a
110 100 0
70 W&-
40' 30
1
0-
13 TtoUaMi U U U 3 7.0 73
Centre Analytical Laboratories, Inc.
Page 57 of 102 CO 0 3 0
Centre Study No.: 023-063
Figure 5. Chromatogram Representing Control Mallard Membrane for PFOS (Centre ID: 0003802 Blank A, Set: 032601C)
S M U X atoxi
C* MMPclr*nuYAypUaeini
AS. D atai AC. T lM t
n
S9U.0o0k0
93/2Y /01 0llllC 7 |
M adltle*i Pan e fatug F M t B n M il m u iu U i.
ACM I k t t D s M i
M . OSSOtftOt
trimK t tfUtfovtrrt
3*9- SitftA i M*p. M U lU i b tp . taak Ratloi Kxp. A4J. R acial C ap. V a l. Mactot
tts o M U t i v o RTi
1Yaa
S. 04
32 4 .JO
30.0
4.43 0.20 0.01 4.00 4.00 3.00
la c . Typoi M c m U m T iasi At m i
lurt TlMi
tact t i m i
4.44 i 40.0 1.44a*002
4.1? |
4.41 i
Centre Analytical Laboratories, Inc.
Page 58 of 102 000302-
Centre Study No.: 023-063
Figure 6. Chromatogram Representing Control Mallard Membrane Fortified with 10 ppb of PFOS, (Centre ID: 0003802 Spk A Dup. Inj., Set: 032601C)
C eaw ur> U M i o.300
Acq. & '
03/21/01
Acq. TlJMt
93.SC.1C n t
lu M U a i Facto*i R o ll* TIUrtldi
in
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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)
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APPENDIX A
Study Protocol OOP-023-063
(Centre Study No. 023-063) and
Amendments and Deviations
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0003
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CemieProtocol No.OOP-023-063
ANALYTICAL PHASE PROTOCOL
ANALYTICAL PHASE TITLE EXTRACTION OF POTASSIUM PERFLUOROOCTANESULFONATEFROM v n n
MEMBRANE, YOLK AND ALBUMEN FOR ANALYSIS USING HPLC ELECTROSPRAY/MASS SPECTROMETRY
SPONSOR 3MEnvironmental Technology and Safety Services
Building 2-3E-09 PO Box 33331 St. Paul, MN 55133-3331
DATA REQUIREMENTS Analytical Method Requirements
PERFORMING LABORATORY Centre Analytical Laboratories, Inc. (Centre)
3048 Research Drive State College, PA 16801
Phone 814-231-8032
PROTOCOL IDENTIFICATION NUMBER OOP-023-063
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0003O S
Centre Study No.: 023-063
CentreProtocol No. OOP-023-063
Title: EXTRACTION OF POTASSIUM PERFLUOROOCTANESULFONATEFROM EGG MEMBRANE, YOLK AND ALBUMEN FOR ANALYSIS USING HPLCELECTROSPRAY/MASS SPECTROMETRY
TABLE OF CONTENTS '' _________ _____________________________ _____ Page TABLE OF CONTENTS------------------------------------------------------------------ -
1. PURPOSE------------------------------------------------------------------------- -
2. REFERENCE MATERIAI-------------------------------------------------------- -
3. SPONSOR_________________________________________________
4. TESTINGFACILITY---------------------------------------------
4
5. PERFORMINGLABORATORY--------------------------------------------------
6. PROPOSEDEXPERIMENTALTIME-FRAME--------------
4
7. JUSTIFICATIONFORTHE SELECTION OFTHETESTSYSTEM------------4
8. SAMPLEPROCESSING, STORAGE AND IDENTIFICATION-----------------5
9. ANALYTICALMETHOD---------------------------------------------------------
10. EXPERIMENTALDESIGN------------------------------------------------------ 11. PROTOCOLAMENDMENTS ANDDEVIATIONS---------------------------- -
12. RECORDS________________________________________________ _
13. QUALITY ASSURANCE-------- ----------------------------
7
14. DATA AND REPORT------------------------------------------------------------ -
15. ARCHIVE STATEMENT....
8
16. REFERENCES----------------------------------------------------------------------- --
17. PROTOCOL APPROVAL...------- ------------------------------------------------- --
APPENDIX: ANALYTICALMETHOD--------------------------------- ----- ----------- 10
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~ Cetrtffe Study No:: 023-063
CentreProtocolNo.OOP-023-063
Title: EXTRACTION OFPOTASSIUM PERFLUOROOCTANESULFONATEFROM p n r,
MEMBRANE. YOLK ANDALBUMEN FOR ANALYSIS USING HPLT ELECTROSPRAY/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 methodnumber 00M-023-015.
The analytical phase will be conducted under EPA TSCA Good Laboratory Practice standards 40 CFRPart 792 (1).
Z REFERENCEMATERIAL The following analytical standards will be used:
Test Material PFOS
TCR Number TCR00017-46
Purity (%) 97S
Expiration Date 08/31/01
Chemical name and structure of the compoundis presented below.
PFOS Chemical Name: IUPAC Name:
Perfluorooctanesulfonate I-Octanesulfonic acid,l,1,203,3,4,4,5,5,6,6,7,7,8,8,8heptadecafluoro-, potassium salt
CAS Number.
2795-39-3
Molecular Weight: 499 (CtFnS03~)
OIt
CgF 1 7 S"
II
0
Note- The neutral molecule and standard form that PFOS (anion) is derived from ^perfluorooctanesulfonate potassium salt [C.FraSOjK], 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 standaidsAest substances and any prepared solutions must be identified with a unique label or number on the container or cross-referenced to the container.
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600-3Off
Centre Study No.: 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
PRINCIPAL INVESTIGATOR: 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
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hahitaw include bushy pastures, grassy roadsides, farmlands and open woodlands, and they are considered land grazers. Mallard habitats include ponds, lakes and marshes, and they are considered aquatic grazers. The use of these matrices for the study was to determine if FFOS 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 Ma&ard 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 Noprocessing will berequired for the any of the samples.
Farh 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 method. The tnpbKwill be kept isolated from thetest substance during storage.
Sample receipt and storage location and conditions during the study will be Hnmrnwtted- All samples and any resulting sample extracts will be identified with a unique label or sample number. Such identification will be either on the containeror cross-referenced to the container.
9. ANALYTICALMETHOD
All samples will be analyzed according to the analytical method tided "Determination of Perfluorooctanesulfonate in Egg Membrane, Albumen, and Yolk by LC/MS/MS", Centre method number OOM-023-015
10. EXPERIMENTAL DESIGN
Sample obtained by Wildlife International Ltd. have been shipped to Centre Analytical Laboratories, Inc. for analysis. The are no known contaminants expectedin the test system that would affect the results ofdus study.
Only tho"* 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 OOM023-015.
Methods to control bias will include assay of untreated control samples, fortification of untreated control samples to obtain recovery data, and replicate
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Centre ProtocolNo. OOP-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 falffltated 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 flrVr^rc sample remains after extraction. Foreach matrix, 0.1 g samples will be weighed and dried in an oven at - 105C for - 12-14 hours. Percent moisture for each 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 Direct' and Sponsor Representative. Planned changes to this protocol or to the analytical method will be mack in writing as an amendment and approved by the Principal Investigator the Study Director, and the Sponsor Representative. Any
deviations will be appended to this protocol and included in the final report.
1. Protocol amendments: Planned changes to die approved protocol shall be
documented by amendments that clearly describe the change, justification for
the change, and impact on the study. Amendments will be signed and dated by
thePrincipal Investigator, StudyDirector, andSponsorRepresentative. 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 aid
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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00310
Centre Study No;: 023-063
Cintre Protocol No. 0QP-O23-063
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 m il additionally include the concentration (e.g., pgtal, ng/mL, ppb, ppt, etc.).
Each 4ta 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 fust 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 dat 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 tire Study Director.
14. DATA AND REPORT
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 1 Analytical and statistical methods used
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O O O tiJjl
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Centre Protocol No. OOP-023-063
3. materials identified by name, lot, purity, and other characteristics
4. Name of performing laboratory and analytical start and termination dates 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 finalize 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 GLP 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 mwictnvmr 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: StudyDirector Copy. Centre Archives
15. ARCHIVESTATEMENT
Study records to be maintained: Records to be maintained for the study include all raw data, observations recorded during the conduct of die study, documentation, chromatograms, specimen tracking information, and study related correspondence. This a description of equipment used during the conduct of the study. All characterization darn and any shipping records shall be retained.
rwviim^.t frh iw 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 Pan 792.
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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
rL -- ULr Naomi Lovallo, PhD. Quality Assurance Auditor
Principal Investigator, Centre
I=*Iqi Kate
'S tlu ffe r Scientist
Facility Management, Centre
S tu d y D ir e c to r , W ild life In te r n a tio n a l, L t d .
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Centre Study No.: 023-063
Ctare Protocol No. 00P-O23-O63 APPENDIX: ANALYTICAL METHOD
A. "Dctennination of Perfluorooctanesulfonate in Egg Membrane, Album s anj ' Yolk by LC/MS/MS", Centre method number 0OM-O23-O15
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C e n tre M ethod N o:
3 3 . EquipmentandSupplies
E Q U IP M E N T
B a la n ce , 5 p in e analytical B a h y i [ p lace
M in i B ead B eatcr-t W o sM ctien shaker B e n d H o p centrifu ge M in t-ce n trifu g e -B 01 SO m L disposable polypropylen e ce m rifu g c'tu b g S tain less ste d b a d s ( a t f 11079) M jc n x a u r ifu g e tubes D isp o sab le pipettes, te a tubes e tc 1 3 -tn . disposable polypropylene ce n trifu g e tu b a
W hatm an1" A n otcj)1" fib e r (eat # 6809-1022) m il Eppendorf" tn fao ejn trifu ge lub es (cat# 2 2 -
36-335=2) 2 -m L d e a r H E ljC v ia i k it (bat # 3111 -3 400) . S ta n d a ri lab equipm ent (d ass A pipettes and vo tm en ie fla s k , graduated cy B n d en , e tc .) IX V M S M S and H F L C system s .
-SQUgjp
M etder M ettler B io % e ftn d a co
EC VW R VW R
VW R VW R VW R VW R VW R
HP
* * scrib e d i , _S e a ia n 4.a i
liais _ L I d Older to awed contamination, lb use of
,_, * *
. h igh ly rccom m eaded (co n ia in e n , tu b a , p ip ettes, e tc.)"- TM lt" c "
- Z T eflo o o r tflc o Jm e d e n n u in e a o r eq uipm ent, n d tK fin * t*ru- r '
H P 1 C v ial caps,should m a be u sed . .
" " " I W bao-Iincd
: ' 4. It is n eessasy to ch eck the so lv en ts (m ethm oQ fo r A ,
eontam inants by L O M S / M S b e fo re u se. Certain lo t
been found to b e 'im ju io b le fo r u se . .
" U tnhes have
s. -Ihe deposable dneropipeues or pipettes to aliquot standard J k -o L
and gbaptetUnet standards and samples forcstracrioo. * Wnno,u
" 6. .Equivalent mataialsmay be.substimted fcr diosc - 4 - . .
method Hw tw r. d* use' of carbon `o n SunefaTtT
recommended.
Wtojiy
3.4.SOUJI1QI.
'
L SO mM anwnonintn acetate soitnion:' Dissolve 3ZS t of -
'
acetate in I L ofASTM type I water. Store in an a p p r a ^ l S * ""
'm antcap eaneforiiptolyear.
. W D Pna* I t e bottle
1 ' 2 inM ammon im acetate solution: Dilute 40 mL of . ----------, -------------, jolutioo in a liter of ASTM type I
w
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CentreMethod
p im c A . Store is an appropriate ( la s b o n k at room u I-je a r.
H * g T h e vohnnej ahrm p are provided fo r ja d a n c t; a h e a J
m ay be prepared.
n TM .ve W hane*
3 i. RtHARAHt OF STOtt, FORTIRCAUDH, AMD CaLbRATION
* soumoNs '
'!
. *I
are prepared fo r tw o p o p . T h ey are toed to fo n fy - J T t T if a g a in e a a tju ro l ttstw ety an il to ca E b n ie t ie
. th e a b s o te ^ K to " 0 0 ^ * " *00S a * e e o n t a 'n>e-a a l f f ? L ^ to to iV d ifa re n n in u in e i T h e t a k t e d ie d b elo w are
#' *<y_ ' - rirrV
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F R 1WMfaL-W " 0 * 1 w lO Jtn to f
^ i ^ t i T . KKMnLwtanemc ta k . .Store thu sod bdon fm
' '! ^ S b o ( U b ) i o * * r ^ " w ^ t e * , ^ P ^ o i '
- K p e ttn i - > d - o f 1 p s ^ L M ock
- l l- io4 t ^a sb u of m ^ S, e t r i e f l a s f c B rio * o p lo .w h r n *
mrrhanol
with-'
V - - ' ftffiM iaJslaisa - Kpette lOD mL of the 1J>iitfm) 'K aTonl. w han* Bk nd 6dn| up to
f tt'in--.withtnettanol.
,n W D H W _t ^
w ith m e g a -
.
4. ;n fnriiF-^ gaytal wlatioio 0" ITSnnLLDPEbottle) hi a
.tfjrcontiofl.
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l i l r*sniitinSm&r&
,,___ .. -ribS/MS caBboioo sandatds in methanolm Anion of the
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h je 9
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' C oin M eth o d N o : OOM-9 & 4 1 5
* . ' **
T h is a a typ ical exam ple; add itiona l c o n c a s s io n s m zv k .
*
7 " P*s>*rcd as
, b aciai C o n e . (u s/m Ll
0.1
0.1 ,0.1 `
; 0.01
. oxi oxi
' 100 1 100 j 100
I 100 015 100
' S u r e anll calib ratio n stan d sd soitnationos (in 125-tnL L D p g
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0.001
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.
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refrigerator S 2 * 0 to "C fc r 1tau xbm m period o f fi m onths Go b S * * ^ ^ . preparation.
4. MEIBOD
.4 .1 .'R w D ia g r a m
H ie flo w diagram o f th e., m ethod is given b e lo w ,'fo llo w e d h v .
descriptio n o f each step .
.
,
. W eigh 1 g o f m a in * (O dg fo n n em b ran e},';F o n iiy*jf n eed ed -
- '
. . . . Extract w ith M eO H
'
. Carbon dean np (except mem brane)
.' .
..
filte r
l
D fludon
. '- l '
' .
` L O M S/M S ansdjs ir . .
`
-
*
-
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.*
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` A U sam p les are w a iv e d A n cenan d w fll be kept fro m '(b e lo w -1 0 , q ^
-
4 s .S A M P lE E C T R .tC n O N . .
N O T E A A e g a m p le n ia in c a were separateti prior to arrival a r n t r c
. C aacA inlytialU ocaarics.be.S aidyIIB M U
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I a tre M ethod N o : D O M -O B ^ -
43.1 EfS Mawrant
L A llo w sam ple to d a w .
bu W d r itO Jt fc Q JtB r i o f a m p le t o o the w eight to the nearest 0.0001* . r b n ifr o a n e m ri
p o in t fo r d a ezn riiB tio o 'o fn reth o d reco tety .
. ._ 8 c "<
* 7^ * d r is -
A d d 1 m L afM e O H a n d t
------ -------- . * * * * * .
h otn ogcnit t w ith bead b e ate rset stp o sitio n 7 ( - 2S B n n '< / y f t ***
C e a tr iftg e the tubes w ith m ia i-ce a iriiin g e 1 0 0 0 n a f o r * ! " " " " v
then ca re M ly transfer supernatan t to a n c w p .m r ^ n [|
fflte tin jiH th W h a n ia n TM A a o c o p 1 0 fflte rlfn e e e * s a i/ ,oe iTM ,i ^
ethanol ^ i ! % ^ l e ' m c m p i p ^ ^ ^ .
e. -
Sto ft*
re th fttn u
e e
test re-d
o ih
f the itk H
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x r
tract re -in
is je c
a re tio n
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untreated eouttol sam ples at th is p o in t-fo r d e te n a n a tio n f
.
tp co v e ry .
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.
d . T ran sfer about S ad. of extract in to a U -m L d isp o sab le 'm ii,;
w a tn fty n t e a d d 0 -5 * carbon , rep lace S d tig h ty , arid s h a k e ^ S f a
e . linato thae m10b-asdsi'tdIibspr-o2sabnled nsaptersin, egaer,ebftatlnTeyltrcaonnsafeeore-Zd O nrrn L, tr f ih e ia n m i.
A n otop" 10 filte r. F ilte r sam ple in to tw o H P L C y ia fe , ^
. ' and one fc r storage in a refrigerator at ap p ro xim ately 2 * C to i t r r ^ f 80"
re-dthm oo er re-in jectio n .
<- to rn ito re
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Page 21 of38
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4 .4 . A n a l y s e b y L C / M S / M S
Ct0in M e th o d N o : OOM -O23415
4.4J: ICMSIMSSystem andOperatingCendtdons(Ttaialonqi j
M a S p e c . P E C E C A H 3000 B io m aleeu lar
bustes
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Potw er M acintosh G3 *
P E S e ie x A M ljs tU " W indow s N T
B ew ies P ackard (H P JSeris U 0 0 H PQ ssPenp. H P V acan o D e fisse B P A m o m o p Is
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lhe H P LC p a & n t fa w rier to opomixj
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a C i , e tc.)
b a sm o n h n sfc '
A n a h e e . . U S T ra n siti M om tw eA
P F O S . p riv e
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batch etm obile phase, etc.
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Centre Analytical Laboratories, Inc.
Page22 of3 Page 83 o f 102
0 0 0 3 ;'
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Centre Study No.: 023-063
eoe Protocol No. OOP-023-063
V
' ' CentreMethodNotOOM.^.
4.43. ExampleToneFtParamtUn . *
T h e fo flo w in j valu er are provided as an
from io sg e m e a to im m iin e a t Also, these vetoes m ay be d a m Z i
to tim e ia o rd a 'to o p c tim z e fw |ie a te s tie a iid n r r.
^
**** tim e
T h e n u sip e c tro m d e r attu n ed m in i * O -S u ffa L fF O S so ta tio ,,
dthtdou at the o d e so la d o a h i m ethan oL T h e m t a it a h
***
* T " connector) a t 10 p L to m in to a 0.2 mUum stream , * * * ^"n* *
acrpeohdnsdtaiOaijrotfon%ediBteefdthHe ptiadraeinitdi6o0n%ta2d.mthMenatmnamed*(matmfaaZclafijJ,ivv*!l*efl****>*,e***
the im m in en t is tuned, the o p tim ized p aram etea a rt s a v ^ 0TM 8 * '
T h is ta ie file -is then used d a rin } ro u tin e an aly sis. T h tan in * * * *lu i,e *****
h e repeated,as necessaty to ensure o p t n a l sen sitiv ity .
P fssth jre n a y
v-onrai
* E -Jn sja a y .
O R -O rific R i*3 -F o cn sR in j
Q O Q u ad .Q g o d O ffe r K J-h a e rq d d 1 le ` S T E tn h h ief ` R O l-Q o a d l R o d O fib t _
IQ 2-Interquad2 1eM /
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-S T 3 -S m h h ies. ' R 0 3 -jQ e a d 3 ro d offset D F - B iD e fle e n o o P late C E M -O u n u id B ce m a i M u ltip lie r
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'4.43. CeibratwnPrtctdura-
a.
'b jm the sam e w taam (betw een 10 to 20 p L ) 6f (prepared in M eO H ) in to the L C / M S / M S .'
each
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.
b . th e Sneer. Ux tin te d standard cu rves 1er
T[
standani a n v e r ere je n e n te d fo r each se t'b y B asar regression u s in g - ^
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fo rtific3niaOiDnJ a i 10, and 2J 0 n j/ i (ppb) fo r tbe
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h . Fortificatio n recoveries w ithin 40 to 1 3 0 * are > ~ t t -nic . .
;
it the L O Q lev el. Recoveries b e t a 70 a n g - _ . ` o n i" tio w
liforrificaaons at k v d a.g riate r d u L O Q .
io r
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,,a
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L Sam ples in w hich bo peaks a t (acted i f the tetcarioo f la g w d be repotted a t M D fa st i f a .^ |I||||[| | nalyre o e a b a ra detected at die h w tw ao ciiitm h itg .w p ^ ^ r t ^ ^ in w hich
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' * / *
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E n t J jijH ia - f e je e t a standard s o h m o o o o th e L O M S ftC e o n e sp o n ifin .i-iK . esd atated L O Q (10 ppb I Q trerprivafent to a a t a o d a r t 'g t e m ^ ^ " ' ^
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' obtain a lin ear regression w idi a bo eflleieal o f detenaiiaim a <r*) o f M j - J l
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' analyzed w ith standards ia n ip e tm d . -
. "***
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C ffl& e M eth o d N o : O O M -023^15
4 .6 . T im e R e q u ir e d k r k a l y s s A le t. t f M a m p le * cm b e taken threnjfa b e e a r e d ip proj m alely d u e b o a s by- one p e n .' l e L O M S / M S l* * T lu re *n S 2n d b x n d id a n q ile s lv iS u k e ip ;x Q z ia n te iy 6 b o Q n .
s- : CALCULATIONS.
5 .1 A k a l y i e F o u k k Cafadae (he imount of analyte tbond (a nttoh. baaed 1 , r< .. 'aandudam* generatedbytbeMreQae 10ft* pnisnin ejEonrektU* * " . J^attaoL* A n th te foim d fae/m LI c (peak B O - in e ig n tl
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C en tre M ethod N O O M -O ZM 15
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T h e n ere do n u s a l h e z sd s etso d ated ridi th is m ethod. T h e a i u w
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T a b le fc Sm anrj o fR e c o r tr ie s fo r P R O S In Q u a il E j j
S a ijfc
F a tlm T
________ ________fadrt
KKBftvIteftBtafcA eeen n iW tB fad cr. e o o n n ita tta w c oocmS(*A . a n sisfts
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Page 34 of 38 Page 95 o f 102
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Page 37 of 38
Page 98 o f 102
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Centre Study No.: 023-063
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Page 38 of 38
Page 99 o f 102
0003
Centre Study No.: 023-063
3048 Research Drive, State CoUt|e, PA 16801.
Phone: (814)231-8032, Facsimile: (814)231-1253
PROTOCOL AMENDMENT Amendment Number. 1 Effective Date: 3/23/01
Page 1 of 1
Centre Study Number 023-063
Centre Protocol Number OOP-023-Qfit
DESCRIPTION OF AMENDED SECTION
1. Tide
2. 8AnalyticalMethod4.4.4.f.
AMEMBEDJQ
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.
Study Diifrpercftrotrf .S^igonathuimre
G |u |21
Date
CentreQAUReview Aifh.
--________ February12. 1998/1
Centre Analytical Laboratories, Inc.
Page 100 o f 102
Centre Study No.: 023.-063
304* lUsmrdiPriTt, Stote Cotte. VA l 0 I.
Phone 814Q31-Sn32. Fatrimiu, /a n w n .i?
PROTOCOL AMENDMENT Amendment Number. 2 Effective Date: 3/26/D1
Page 1 of 1
Centre Study Number 023-063
Centre Protocol Number OOP-Q2 3 -Q63
DESCRIPTION OF AMENDED SECTION
1. Section 9: Analytical Method 2. Section 9: Analytical Method 3. Section 10: Experimental Design
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 IH 2O and patted dry.
2. On Page 24 Section 4.4.4.d, remove: One ASTM TypeI water 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.- 12-1'4 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 I HjO used.
3. Toaccount forsampleavailability.
IMPACT ON THE STUDY
1-3. No negative impact on the study.
Date
Centre Analytical Laboratories, Inc.
Centre QAU Review
February12. 1998/1
Page 101 of 102
GOOGAJ
Centre Study No.: 023-063
3048 R esearch D rive, S tate C edete, PA 10801.
Phone: (814) 2314032, Facsim ile; (814) 231-1253
PROTOCOL DEVIATION
Deviation Number 1
Date ofOccurrence: (1)03/27/01,(2)04/02/01
Centre Study Number 023-063
Centre Protocol Number OOP-023-063
D E SC R IP T IO N O F D E V IA T IO N
1. 58 Analytical Method 4.4.4.f: Quantified Centre sample 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% far Centre sample 0003800 Spk A and its duplicate injection in Set 033001C
ACTIONS TAKEN U .. imendmem ioued. SOP revision, eie..
1-2. Protocol deviation issued.
Recorded By/Date:
IM P A C T O N T H E S T U D Y
1. No negative impact because die areas were < 5% greater than the highest standard.
2. No negative impact because Spk B gave acceptable recoveries and sample results reported in the set were only for confirmation.
estigator Signature
k fel Date
Study Director SSigemnaatnuarre f'tSfiWit,
2 11 V Date
O t l/o S
D ate
CAL QAU Review. f f k l i b i l i
F tb e im y 12,1998/2
Centre Analytical Laboratories, Inc.
Page 102 of 102
o c o iv 6
