Document K31X0kg1GQ8Zz4XRzd85wyBw

AR226-1740 STUDY TITLE Extraction of Potassium Perfluorooctanesulfonatefrom Egg Membrane, Albumen, and Yolk for Analysis Using HPLC-ElectrosprayMass Spectrometry DATA REOUIREMENTS 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: OOP-023-063 Centre Study Number: 023-063 Total Pages: 102 centraStudy No.:023663 GOODLAB0RATORYPRACTICE CKlMPLIANCE S T M " T Cmtm Study Numbcr 023-065, antic14 `'Exmaion of Potassium Petfluomootanesulfonate h m Egg Membrene, Albumen, and Yolk for Analysis Using HPLC-ElecmqmyMass Spectrometry," conducted for 3M Environmental Technology and Safety Services, was performedin compliance with US EPA TSCA QoodLaboratory Practice Standards (40 CFR Put 792) by Centre Analytical Ldboratorica, Inc with thc fallowing exceptions. 1, $792.63. Mainmnaua and Calibration of Equipment. No equipment SOP was in place for thc btad beaterueedin the extraction at the tinae of use. L P- Sam Gallagher 2&)-- Study Director Wildlife Intunational, LEd. CentreAnalytical LaboratdeB,Lno. P q c 2 of 102 Centre Study No.: 023-063 OUALITYASSURANCE 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-ElectrosprayMass 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. FinalReport Review Date Inspected 3/30/0 1 3l30lO 1 Date Reported to Date Reported to Centre Study Director and Manavement Sponsor Management 41910 1 51710 1 41910 1 51710 1 4/5-9101 4117-18/01 4/18/01 412610 1 5/7/01 5/7/01 6/28/0 1 91610 1 91610 1 Date Centre Analytical Laboratories, Inc. Page 3 of 102 CentreStudy No.:023463 This repait, for Centte SWdy Number 023-063.is B true anQ cornplate mpresenration of the raw data for the study. Submitted by: Centre AnalyticalLaboratories,I~c. 3048 R-h Drive State College, PA 16801 (814) 231-8032 Principal Investigafor,Centre: Crntre AnalydcB bboratdw, bc. Cenrsc Analytical Laboratories,Inc,Facility Management: President cenm Analytical hboratories, hc. Study Director Date Wildlife International,Ltd Sponsor Repremcative Page 4 of 102 Centre Study No.: 023-063 STUDY IDENTIFICATION "Extraction of Potassium Perfluorooctanesulfonate from Egg Membrane, Albumen, and Yolk for Analysis Using HPLC-ElectrosprayLMass Spectrometry," CENTRE PROTOCOL NUMBER: OOP-023-063 CENTRE STUDY NUMBER: 023-063 TYPE OF STUDY: Analytical SAMPLEMATRIX: 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 Wildlife International, Ltd. 8598 Commerce Drive Easton, MD 21601 TESTING FACILITY: Centre Analytical Laboratories, Inc. 3048 Research Drive State College, PA 16801 ANALYTICALPHASE TIMETABLE: Study Initiation Date: Experimental Start Date: Experimental Termination Date: Study Completion Date: 03/26/0 1 03/26/0 1 04/02/01 09/06/01 Centre Analytical Laboratories, Inc. Page 5 of 102 Centre Study No.: 023-063 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 Aqmand Ling Ling Liu Yu Qiong Tan Xiaoming Zhu Lawrence Ord Rickey Keller Shawn Robb - Title Scientist Scientist Technician Technician Technician Technician Sample Custodian Sample Custodian Sample Custodian Centre Analytical Laboratories, Inc. Page 6 of 102 . Centre Study No. 023-063 TABLE OF CONTENTS PaJg TITLEPAGE ...................................................................................................................... 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 6.6 Quantitation 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 Centre Analytical Laboratories. Inc. Page 7 of 102 . Centre Study No. 023-063 LIST OF TABLES Table I. Summary of PFOS in Reagent Blanks........................................................... 23 Table II. Summary of PFOS in Mallard Membrane Blanks ......................................... 24 Table III. Summary of PFOS in Mallard Albumen Blanks............................................ 24 Table IV. Summary of PFOS in Mallard Yolk Blanks .................................................. 24 Table V. Summary of PFOS Recoveries in Mallard Membrane................................... 25 Table VI. Summary of PFOS Recoveries in Mallard Albumen ..................................... 25 Table VII. Summary of PFOS Recoveries in Mallard Yolk............................................ 26 Table Vm. Summary of PFOS for Week 1Mallard Membrane Control Samples...........26 Table IX. Summary of PFOS for Week 1Mallard Membrane Samples at 6.2 ppm ai....................................................................................................... 27 Table X. Summary of PFOS for Week 1 Mallard Membrane Samples at 17.6ppm ai..................................................................................................... 27 Table XI. Summary of PFOS for Week 1 Mallard Albumen Control Samples.............28 Table XII. Summary of PFOS for Week 1 Mallard Albumen Samples at 6.2 ppm ai ....28 Table XIII. Summary of PFOS for Week 1 Mallard Albumen Samples at 17.6ppm ai ..29 Table X N.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 ai ...........30 Table XVI. Summary of PFOS for Week 1 Mallard Yolk Samples at 17.6 ppm ai .........30 Table XVII.Summary of PFOS for Week 6 Mallard Membrane Control Samples..........31 Table XVm.Summary of PFOS for Week 6 Mallard Membrane Samples at 6.2 ppm ai..................................................................................................... 31 Table XIX. Summary of PFOS for Week 6 Mallard Membrane Samples at 17.6ppm ai..................................................................................................... 32 Table XX. Summary of PFOS for Week 6 Mallard Albumen Control Samples.............32 Table XXI. Summary of PFOS for Week 6 Mallard Albumen Samples at 6.2 ppm ai ....33 Table XXII.Summary of PFOS for Week 6 Mallard Albumen Samples at 17.6ppm ai .33 Table X x m. Summary of PFOS for Week 6 Mallard Yolk Control Samples...........34 Table XMV . Summary of PFOS for Week 6 Mallard Yolk Samples at 6.2 ppm ai ..34 Centre Analytical Laboratories. Inc. Page 8 of 102 . Centre Study No. 023-063 Table XXV. Summary of PFOS for Week 6 Mallard Yolk Samples at 17.6 ppm ai ....35 Table XXVI. Summaryof PFOS in Quail Membrane Blanks .................................... 35 Table XXVII. Summary of PFOS in Quail Albumen Blanks ........................................... 35 Table Xxvm.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 X X X I.Summary of PFOS Recoveries in Quail Yolk............................................. 37 Table X X X I T .Summary of PFOS for Week 1 Quail Membrane Control Samples ..........38 Table Xxxm.Summary of PFOS for Week 1 Quail Membrane Samples at 6.2ppm ai.................................................................................................. 38 Table XXXIV .Summary of PFOS for Week 1 Quail Membrane Samples at 17.6ppm ai................................................................................................ 39 Table XXXV.Summary of PFOS for Week 1 Quail Albumen Control Samples ............39 Table XXXVI. Summary of PFOS for Week 1 Quail Albumen Samples at 6.2 ppm ai...40 Table XXXVlI.Summary of PFOS for Week 1 Quail Albumen Samples at 17.6 ppm ai .............................................................................................. 40 Table xxxvII[.Summary of PFOS for Week 1 Quail Yolk Control Samples................41 Table XXXIX.Summary of PFOS for Week 1 Quail Yolk Samples at 6.2ppm ai .........41 Table XL. Summary of PFOS for Week 1 Quail Yolk Samples at 17.6ppm ai .............42 Table XLI.Summary of PFOS for Week 6 Quail Membrane Control Samples...............42 Table XLII.Summary of PFOS for Week 6 Quail Membrane Samples at 6.2 pprn ai .....43 Table XLIII .Summary of PFOS for Week 6 Quail Membrane Samples at 17.6pprn ai ..43 Table XLIV .Summary of PFOS for Week 6 Quail Albumen Control Samples...............44 Table XLV.Summary of PFOS for Week 6 Quail Albumen Samples at 6.2 ppm ai .......44 Table =VI . Summary of PFOS for Week 6 Quail Albumen Samples at 17.6 ppm ai ....45 Table XLVII.Summary of PFOS for Week 6 Quail Yolk Control Samples .................... 45 Table XLW.Summary of PFOS for Week 6 Quail Yolk Samples at 6.2 ppm ai ..........46 Table IL. Summary of PFOS for Week 6 Quail Yolk Samples at 17.6ppm ai...............46 Table L. Summary of Moisture Determinations for Mallard Membrane Samples.......47 Table L,I. Summary of Moisture Determinations for Mallard Albumen Samples.........48 Centre Analytical Laboratories. Inc. Page 9 of 102 . Centre Study No. 023-063 Table LII. Summary of MoistureDeterminations for Mallard Yolk Samples................49 Table LIII. Summary of Moisture Determinations for Quail Membrane Samples ..........50 Table LIV. Summary of Moisture Determinations for Quail Albumen Samples.............51 Table LV. Summary of MoistureDeterminations for Quail Yolk Samples.................... 52 Centre Analytical Laboratories. Inc . Page 10 of 102 Centre Study No.: 023-063 Figure 1. Figure 2. Figure 3. Figure 4. Figure 5. Figure 6. Figure 7. LIST OF FIGURES Page Typical Calibration Curve for PFOS.............................................................. 54 chromatogram Representing a 0.1 ng/mL standard for PFOS ...................... 55 Chromatogram Representing a 1.0ng/mL standard for PFOS ...................... 56 Chromatogram Representing a Reagent Blank for PFOS (Centre ID Reagent Blank A, Set: 032601A)................................................. 57 Chromatogram Representing Control Mallard Membrane for PFOS (Centre ID: 0003802Blank A, Set: 032601C)............................................................ 58 Chromatogram Representing Control Mallard Membrane Fortified with 10 ppb of PFOS, (Centre ID: 0003802 Spk A Dup. In$, Set: 032601C)............59 Chromatogram Representing Mallard Membrane Sample from Week 1 at 6.2 ppm ai pol-0263-20825) (Centre ID: 0103092, Set: 032601C).........60 Centre Analytical Laboratories, Inc. Page 11 of 102 Centre Study No.:023-063 LIST OF APPENDICES Page Appendix A Study Protocol OOP-023-063 (Centre Study No. 023-063) and Amendments and Deviation.....................................................................61 Centre Analytical Laboratories, Inc. Page 12 of 102 Centre Study No.: 023-063 1.0 SUMMARY Centre Analytical Laboratories, Inc. (Centre) extracted mallard and quail egg membrane, albumen and yolk samples for the determination of perfluorooctanesulfonate (PFOS) according to protocol OOP-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 f standard deviations for PFOS in mallard membrane, albumen, and yolk samples were 91% f 6%, 91% f 16%, and 93% k 13%, respectively. The average percent recoveries f standard deviations for PFOS in quail membrane, albumen, and yolk samples were 102% f 11%, 85% f 5%, and 96% f 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 Perfluorooctanesulfonatein 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. Centre Analytical Laboratories, Inc. Page 13 of 102 Centre Study No.: 023-063 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 u> Mallard Membrane 0003802 Mallard Albumen 0003800 Mallard Yolk 0003799 Quail Membrane 0003 79 8 Quail Albumen 0003796 Quail Yolk 0003795 On February 23, 2001, 308 quail egg samples were received. The samples were stored frozen until they were logged in by Centre personnel on February 26, 2001 and stored frozen. On February 24, 2001, 194 mallard egg samples were received. The samples were stored frozen until they were logged in by Centre personnel on March 1,2001 and stored frozen. Each egg sample received was divided into membrane, albumen, yolk, and shell. Each component of the egg was given a unique Centre ID at the time of log-in. Although all of the egg samples were received at Centre, only those designated by the sponsor were extracted and analyzed. Sample log-in and chain of custody information can be found in the raw data package associated with this study. Storage records will be kept at Centre Analytical Laboratories, Inc. and a true copy of the storage records can be found in the raw data package associated with this study. 5.0 REFERENCE MATERIAL 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. ComDound PFOS Centre Control No.. TCR No. Purity (%) ExDiration Date 00-023-042 TCR 00017-46 97.9 08/3 1/01 Centre Analytical Laboratories, Inc. Page 14 of 102 Centre Study No.: 023-063 The molecular structure of PFOS is given below. PFOS Chemical Name - Molecular weight = 0 Peffluorooctanesulfonate 499 (C8F17S03-) Note: The neutral molecule and standard from which the PFOS (anion) is obtained, is potassium peffluorooctanesulfonate[ C ~ F I ~ S O ~mKo]le,cular weight 538. 6.0 DESCRIPTION OF ANALYTICAL METHOD Analytical method entitled "Determination of Peffluorooctanesulfonate in Egg Membrane, Ablumen and Yolk by LC/MS/MS" was used for this study. 6.1 Extraction Procedure Egg membrane A 0.1 g sample was used for the extraction procedure. One milliliter of methanol was added to the sample along with three stainless steel beads. The sample was then placed - - on a bead beater set at position 7 for 5 minutes. The sample was then centrifuged on a mini-centrifuge at 2000 rpm for 2 minutes. Each sample was filtered and analyzed by LC/MS/MS electrospray. Egg albumen and yolk A 1.0 g sample was used for the extraction procedure. Twenty-five milliliters of - - methanol were added to the sample. The sample was placed on a wrist-action shaker for 15 minutes and then centrifuged at 2000 rpm for 10 minutes. A five milliliter aliquot of the sample was taken and 0.5 g of carbon was added. The sample was shaken then allowed to sit. Each sample was filtered and analyzed by LC/MS/MS electrospray. 6.2 Preparation of Standards and Fortification Solutions Standard solutions were prepared on March 14, 2001 as specified in Centre Analytical Laboratories' protocol 00P-023-027. An individual stock standard solution of PFOS was prepared at a concentration of 100 pg/mL by dissolving 10 mg of the standard (corrected for purity and salt content) in methanol. From this solution, a 1.0 pg/mL fortification standard solution was prepared by taking 1mL of the stock and bringing the volume up to 100 mL with methanol. Centre Analytical Laboratories, Inc. Page 15 of 102 Centre Study No.: 023-063 The 0.1 pg/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 Conc. (pg/mL)' 0.1 0.1 0.1 0.01 0.01 ' 0.01 of PFOS Volume (mL) 5 2 1 5 2 1 Diluted to (mL) 100 100 100 100 100 100 Final Conc. (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 f 2OC) 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 time of PFOS was 4.4 was min. accomplished by LC/MS/MS electrospray. Peaks were detected in the control matrices The retention corresponding to the analyte retention time, but the amounts detected were only significant enough to alter the results in several of the recoveries reported and the rest were less than the lowest calibration standard (0.0001 pg/mL). 6.4 Instrument Sensitivity The smallest standard amount injected during the chromatographic run had a concentration of 0.0001 pg/mL of PFOS. Centre Analytical Laboratories,Inc. Page 16 of 102 Centre Study No.: 023-063 6.5 Description of Instrument and Operating Conditions Instrument: PE SClEX API 3000 Biomolecular Mass Analyzer Interface: SCIEX TurboIon Spray Liquid Introduction Interface Computer: Dell Ultrascan P1110 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 Co1umn:Genesis CS(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 mUmin. Time %A 0 60 40 1.o 0 100 7.0 0 100 7.5 60 40 11.0 60 40 Ions monitored Analyte PFOS Mode negative + Transition Monitored 499 99 Approximate Retention Time (min) 4.40 Tune File Parameters Controls IS-Iospray DP-Declustering Potential FP-Focusing Potential EP-Entrance Potential CE-Collision Energy CXP-Collision Cell Exit Potential Centre Analytical Laboratories, Inc. Set -4200.0 -5 1.O -230.0 10.0 -70 -5 Page 17 of 102 Centre Study No.: 023-063 DF-Deflector CEM-Channel Electron Multiplier Gas Flows Nebulizer Gas Curtain Gas Collision Gas TIS Temperature 6.6 Quantitation and Example Calculation 300.0 2800.0 - Set 12 14 4 350C Ten microliters of sample or calibration standard were injected into the LCMSMS. The peak area was measured and the standard curve was generated (using l/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. Ecruation 1: Analyte found (ng/mL) = (Peak area - intercept) slope Equation 2: Analyte found (ng/g) = anal. found (ndmL) x FV (mL) x DF sample wt. (8) Where: FV = Final Volume DF = Dilution Factor For samples fortified with known amounts of PFOS prior to extraction, Equation 3 calculated the percent recovery. Euuation 3: Recovery (%) = (anal. Found (nglg) - 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: Centre Analytical Laboratories, Inc. Page 18 of 102 Centre Study No.: 023-063 peak area intercept slope dilution factor ng/g added (fort level) avg. amt in controls final volume sample weight 3369 97 1 6700 1 10 ng/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) = 13369-9711 6700 = From equation 2: Analyte found (ng/g) = 0.36 ng/mL J0.36ndmL x 25 mL x 1 1.0038 (8) = 8.97 ng/mL From equation 3: % Recovery = (8.97x-ng/ 100 10 ng/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 EXPERIMENTALDESIGN - 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 H20 blank was analyzed. Centre Analytical Laboratories, Inc. Page 19 of 102 Centre Study No.: 023-063 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 11-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 f standard deviations for PFOS in mallard membrane, albumen, and yolk samples were 91% f 6%,91%f 16%, and 93% f 13%, respectively. Individual recoveries for PFOS in the quail membrane, albumen, and yolk samples are detailed in Table XXIX-XXXI.The average percent recoveries f standard deviations for PFOS in quail membrane, albumen, and yolk samples were 102%& 11%, 85% f 5%, and 96%f 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 wet weight basis. Individual results are listed in Tables XXXII-IL.Some of 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 OOP-023-063. Centre Analytical Laboratories, Inc. Page 20 of 102 Centre Study No.: 023-063 10.0 RETENTION OF DATA AND SAMPLES 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. Centre Analytical Laboratories, Inc. Page 21 of 102 Centre Study No.: 023-063 TABLES Centre Analytical Laboratories, Inc. Page 22 of 102 Centre Study No.: 023-063 Table I. Summary of PFOS in Reagent Blanks Sponsor Centre Set Extraction Analysis Analyte na Reagent Blank A 032601A 3/26/01 3126-27101 na Reagent Blank A* 032601A 3126101 3126-27101 na Reagent Blank A 032601B 3/26/01 312710 1 M Reagent Blank A* 032601B 312610 1 312710 1 na Reagent Blank A 032601C 312610 1 312710 1 na Reagent Blank A* 032601C 3/26/01 312710 1 na Reagent Blank A 032701A 3127101 3/28/01 na Reagent Blank A* 032701A 312710 1 3/28/0 1 na Reagent Blank A 032701B 3/27/01 3128-29101 na Reagent Blank A* 032701B 3/27/01 3128-29101 na Reagent Blank A 032701C 3/27/0 1 3/29/01 na Reagent Blank A* 032701C 312710 1 3/29/01 na ReagentBlank A 032801A 3128101 3/30/01 na Reagent Blank A* 032801A 3/28/01 3/30/01 na Reagent Blank A 032801B 312810 1 3/30/0 1 na Reagent Blank A* 032801B 3/28/01 3/30/01 na Reagent Blank A 032901A 312910 1 313 110 1 na Reagent Blank A* 032901A 3/29/01 313 110 1 na Reagent Blank A 032901B 3/29/01 31314/1/01 na Reagent Blank A* 032901B 3/29/01 3131-4/1/01 na ReagentBlank A 033001A 3/30/01 4/1/01 na Reagent Blank A* 033001A 3/30/01 4/1/01 na Reagent Blank A 033001B 3/30/01 41210 1 na Reagent Blank A* 033001B 3/30/01 4/2/01 na Reagent Blank A 033001C 3/30/01 41210 1 na Reagent Blank A* 033001C 3/30/01 41210 1 NQ * Duplicate injection AVERAGE NQ STANDARD DEVIATION 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 of 102 Centre Study No.: 023-063 Table 11. Summary of PFOS in Mallard Membrane Blanks Sponsor Centre Set Extraction Analysis Analyte na 0003802 Blank A 032601C 3/26/01 3/27/01 NQ na 0003802 Blank A* 032601C 312610 1 312710 1 NQ na 0003802 Blank A 032701C 3/27/01 3/29/01 NQ na 0003802 Blank A* 032701C 3/27/01 3/29/01 NQ AVERAGE: NQ STANDARD DEVIATION: NQ Table 111. Summary of PFOS in Mallard Albumen Blanks Sponsor na na na na na na Centre 0003800 Blank A 0003800 Blank A* 0003800 Blank A 0003800 Blank A* 0003800 Blank A 0003800 Blank A* Set Extraction Analysis 032601A 032601A 032701B 032701B 033001C 033001C 3126101 3/26-27101 3/26/01 3/26-27/01 3/27/01 3128-29101 3/27/01 3128-29101 3/30/01 4/2/01 3/30/01 4/2/01 AVERAGE: STANDARD DEVIATION: Analyte NQ NQ NQ NQ 6.03 6.59 2.94 2.62 Table IV. Summary of PFOS in Mallard Yolk Blanks Sponsor Centre Set Extraction Analysis Analyte na 0003799 Blank A 032601B 3/26/0 1 312710 1 NQ na 0003799 Blank A* 032601B 3/26/01 312710 1 NQ na 0003799 Blank A 032701A 3/27/01 3/28/01 9.57 na 0003799 Blank A* 032701A 3/27/01 3/28/01 9.80 AVERAGE: 5.48 STANDARD DEVIATION: 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 of 102 Centre Study No.: 023-063 Table V. Summary of PFOS Recoveries in Mallard Membrane Sponsor Centre Set Extraction Analysis Amt % na 0003802 Spk A 032601C 3/26/01 3/27/01 10 86 M 0003802 Spk A* 032601C 3/26/01 3/27/01 10 87 na 0003802 Spk B 032601C 3/26/01 3/27/01 100 87 na 0003802 Spk B* 032601C 3/26/01 3/27/01 100 81 na 0003802 Spk A 032701C 3/27/01 3/29/01 10 97 na 0003802 Spk A* 032701C 3/27/01 3/29/01 10 94 na 0003802 Spk B 032701C 3/27/01 3/29/01 97 95 na 0003802 Spk B* 032701C 3/27/01 3/29/01 97 98 AVERAGE: 91 STANDARD DEVIATION: 6 RELATIVE STANDARD DEVIATION 7 Table VI. Summary of PFOS Recoveries in Mallard Albumen Sponsor Centre Set Extraction Analysis Amt % na 0003800 Spk A 032601A 3/26/01 3/26-27/01 10 89 na 0003800 Spk A* 032601A 3/26/01 3/26-27/01 10 91 na 0003800 Spk B 032601A 3/26/01 3/26-27/01 99 94 na 0003800 Spk B* 032601A 3/26/01 3/26-27/01 99 92 na 0003800 Spk A 032701B 3/27/01 3/28-29/01 10 107 na 0003800 Spk A* 032701B 3/27/01 3/28-29/01 10 105 na 0003800 Spk B 032701B 3/27/01 3/28-29/01 99 101 na 0003800 Spk B* 032701B 3/27/01 3/28-29/01 99 99 na 0003800 Spk A 033001C 3/30/01 4DJO 1 10 59 na 0003800 Spk A* 033001C 3/30/01 4/2/01 10 61 na 0003800 Spk B 033001C 3/30/01 4/2/01 98474 99 na 0003800 Spk B* 033001C 3/30/01 4/UO 1 98474 99 AVERAGE: 91 STANDARD DEVIATION: 16 RELATIVE STANDARD DEVIATION: 17 * Duplicate Injection LOQ = 10 ppb Centre Analytical Laboratories, Inc. Page 25 of 102 Centre Study No.: 023-063 Table VII. Summary of PFOS Recoveries in Mallard Yolk Sponsor Centre Set Extraction Analysis Amt 9% Na 0003799 Spk A 032601B 3/26/01 3/27/01 10 104 Na 0003799 Spk A* 032601B 3/26/01 3/27/01 10 104 Na 0003799 Spk B 032601B 3/26/01 3/27/01 98 99 Na 0003799 Spk B* 032601B 3/26/01 3127101 98 99 Na 0003799 Spk A 032701A 3/27/01 3128101 10 71 Na 0003799 Spk A* 032701A 3/27/01 3/28/01 10 73 Na 0003799 Spk B 032701A 3/27/01 3128101 97 98 Na 0003799 Spk B* 032701A 3/27/01 3128101 97 94 AVERAGE 93 STANDARD DEVIATION: 13 RELATIVE STANDARD DEVIATION: 14 Table VIII. Summary of PFOS for Week 1 Mallard Membrane Control Samples Sponsor Centre Set Extraction Analysis Analyte E01-0263-20852 EO1-0263-20852* EO 1-0263-20853 EO1-0263-20853* EO 1-0263-20854 EO1-0263-20854* E O 1-0263-20842 EO1-0263-20842* E01-0263-20843 E01-0263-20843* 0103200 0 103200 0103204 0 103204 0103208 0103208 0103160 0103 160 0103164 0103 164 * Duplicate Injection 032601C 032601C 032601C 032601C 032601C 032601C 032601C 032601C 032601C 032601C 3/26/01 312710 1 NQ 3/26/01 3/27/01 NQ 3/26/01 3/27/01 NQ 3/26/01 3/27/01 NQ 3/26/01 3/27/01 NQ 3/26/01 3/27/01 NQ 3/26/01 3/27/01 NQ 312610 1 3/27/01 NQ 312610 1 312710 1 NQ 312610 1 312710 1 NQ AVERAGE: NQ STANDARD DEVIATION NQ NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) 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 Centre Set Extraction Analysis Analyte EO1-0263-20825 EO1-0263-20825* EO1-0263-20846 E01-0263-20846* EO 1-0263-20832 E01-0263-20832* E01-0263-20847 E01-0263-20847* EO1-0263-20848 E01-0263-20848* 0103092 0103092 0103 176 0103176 0103120 0103120 0103 1 80 0103180 0 103 184 0103 184 032601C 032601C 03260 1 C 03260 1 C 032601C 032601C 032601C 032601C 032601C 032601C 312610 1 3/27/01 3/26/01 3/27/01 312610 1 312710 1 3/26/01 312710 1 312610 1 3/27/01 3/26/01 312710 1 312610 1 3/27/01 3/26/01 3/27/01 3/26/01 3/27/01 3/26/01 3/27/01 AVERAGE: STANDARD DEVIATION: 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 Centre Set Extraction Analysis Analyte E01-0263-20856 EO1-0263-20856* E01-0263-20850 E01-0263-20850* EO1-0263-20857 E01-0263-20857* EO1-0263-20851 E01-0263-20851* EO1-0263-20858 E01-0263-20885* 0103216 0103216 0103192 0103 192 0103220 0103220 0103196 0103196 0103224 0 103224 032601C 032601C 032601C 032601C 032601C 032601C 032601C 032601C 032601C 032601C 3/26/01 3/27/01 312610 1 3/27/01 312610 1 3/27/01 312610 1 3/27/01 312610 1 3/27/01 3/26/01 3/27/0 1 312610 1 3/27/01 3/26/01 3/27/01 3/26/01 312710 1 312610 1 312710 1 AVERAGE: STANDARD DEVIATION: 248** 264** 26.5 28.6 78.9 79.0 287** 281** 50.1 49.2 52.1 23.1 * Duplicate Injection LOQ = 10 ppb ** Possible contamination by yolk or outer shell component of sample and as per Sponsor Representative request were not used to calculate the average and standard deviation. Centre 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 Centre Set Extraction Analysis Analyte E01-0263-20852 EO1-0263-20852* EO 1-0263-20853 EO1-0263-20853* E01-0263-20854 EO1-0263-20854* E01-0263-20842 EO1-0263-20842* E01-0263-20843 EO1-0263-20843* 0103201 0 103201 0103205 0103205 0103209 0 103209 0103161 0103 161 0103165 0103 165 032601A 032601A 032601A 032601A 032601A 032601A 032601A 032601A 032601A 032601A 3126101 3126-2710 1 NQ 312610 1 3126-27101 NQ 312610 1 3126-27101 NQ 3126101 3126-27101 NQ 3/26/01 3126-2710 1 NQ 312610 1 3/26-27/01 NQ 3/26/0 1 3126-27101 NQ 3/26/0 1 3126-27101 NQ 312610 1 3126-27101 NQ 3126101 3126-27101 NQ AVERAGE: NQ STANDARD DEVIATION: NQ Table XII. Summary of PFOS for Week 1 Mallard Albumen Samples at 6.2 ppm ai Sponsor Centre Set Extraction Analysis Analyte EO1-0263-20825 E01-0263-20825* E01-0263-20846 EO1-0263-20846* EO1-0263-20832 E01-0263-20832* EO1-0263-20847 EO1-0263-20847* EO 1-0263-20848 E01-0263-20848* 0103093 0103093 0103177 0103 177 0103121 0 103 12 1 0103 18 1 0103 18 1 0103185 0103 185 032601A 032601A 032601A 032601A 03260149 03260119 032601A 032601A 032601A 032601A 3l26lO 1 3126-2710 1 2.49 312610 1 3126-27101 2.75 3l26lO 1 3126-2710 1 7.90 312610 1 3/26-27101 7.89 312610 1 3126-27101 5.67 3/26/01 3/26-27/01 5.79 3/26/01 3/26-27/01 5.76 3/26/01 3/26-27101 5.58 3l26lO 1 3126-2710 1 5.95 3/26/0 1 3126-2710 1 6.29 AVERAGE 5.63 STANDARD DEVIATION: 1.80 * 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 28 of 102 Centre Study No.:023-063 Table XIII. Summary of PFOS for Week 1 Mallard Albumen Samples at 17.6 ppm ai Sponsor Centre Set Extraction Analysis Analyte E0l-0263-20856 EO1-0263-20856* E01-0263-20856 EO 1-0263-20856* E01 -0263-20856 E01-0263-20856* E01-0263-20850 EO1-0263-20850* EO1-0263-20857 E01-0263-20857* EO1-0263-20851 E01-0263-20851* EO1-0263-20858 EO1-0263-20858* 0103217 0 1032 17 01032 17 01032 17 0103217 DUP 0103217 DUP 0103193 0103 193 0103221 010322 1 0103197 0103197 0103225 0 103225 032601D 032601D 033001C 033001C 033001C 033001C 032601A 032601A 032601A 032601A 032601A 032601A 032601A 032601A 3/26/01 3127-28101 312610 1 3/27-2810 1 313010 1 4lUO 1 313010 1 4lUO 1 3l30lO 1 4lUO 1 3l30lO 1 4/2/01 3126101 3126-27101 312610 1 3126-27101 312610 1 3126-27101 3/26/0 1 3126-27101 3126101 3/26-27101 3/26/0 1 3126-27101 312610 1 3126-27101 3/26/0 1 3126-27101 AVERAGE STANDARD DEVIATION: 3550** 3490** 1388** 1369** 845** 771** 24.7 23.9 23.8 23.3 78.0** 69.7** 19.7 19.8 22.5 2.20 Table XIV. Summary of PFOS for Week 1 Mallard Yolk Control Samples Sponsor Centre Set , Extraction Analysis Analyte E01-0263-20852 EO1-0263-20852* E01-0263-20853 EO1-0263-20853* EO 1-0263-20854 EO1-0263-20854* EO1-0263-20842 EO1-0263-20842* EO1-0263-20843 EO1-0263-20843* 0103 198 0103198 0 103202 0 103202 0103206 0 103206 0103158 0103 158 0103162 0103 162 * Duplicate Injection 032601B 032601B 032601B 032601B 032601B 032601B 032601B 032601B 032601B 032601B 3/26/01 3/27/01 NQ 312610 1 3/27/01 NQ 3/26/01 3/27/01 NQ 3/26/01 3/27/01 NQ 312610 1 312710 1 NQ 312610 1 312710 1 NQ 3/26/01 312710 1 NQ 3/26/01 3/27/01 NQ 3/26/01 312710 1 NQ 3/26/01 3/27/01 NQ AVERAGE NQ STANDARD DEVIATION: NQ NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) LOQ = 10 ppb ** Possible contamination by yolk or outer shell component of sample and as per Sponsor Representative request were not used to calculate the average and standard deviation. Centre Analytical Laboratories, Inc. Page 29 of 102 Centre Study No.: 023-063 Table XV. Summary of PFOS for Week 1 Mallard Yolk Samples at 6.2 PPm ai Sponsor Centre Set Extraction Analysis Analyte EO1-0263-20825 E01-0263-20825* E01-0263-20846 E01-0263-20846* EO 1-0263-20832 E01-0263-20832* EO 1-0263-20847 EO1-0263-20847* EO1 -0263-20848 E01-0263-20848* 0103090 0 103090 0103174 0103174 0103118 01031 18 0103178 0103178 0103182 0103182 032601D 032601D 032601D 032601D 032601D 032601D 032601D 032601D 032601D 032601D 3/26/01 3/27-28/01 3/26/01 3/27-2810 1 312610 1 3127-28101 3/26/0 1 3/27-2810 1 312610 1 3127-28101 3/26/01 3127-28101 3/26/01 3127-28101 3/26/01 3127-28/01 3/26/01 3127-28101 312610 1 3127-28/01 AVERAGE: STANDARD DEVIATION: 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 Centre Set Extraction Analysis Analyte E01-0263-20856 EO1-0263-20856* EO1-0263-20850 E01-0263-20850* E01-0263-20857 E01-0263-20857* EO1-0263-20851 E01-0263-20851* EO 1-0263-20858 E01-0263-20858* 0103214 01032 14 0103190 0103 190 0103218 01032 18 0103194 0103194 0103222 0103222 * Duplicate Injection LOQ = 10 ppb 032601D 032601D 0326012) 032601D 032601D 032601D 032601D 032601D 032601D 032601D 3/26/01 3127-28101 3/26/01 3127-28/01 3/26/01 3/27-28/01 312610 1 3127-28/01 312610 1 3127-28101 3/26/01 3127-28101 3/26/01 3/27-28/01 3126101 3/27-28101 312610 1 3/27-28101 312610 1 3/27-28/01 AVERAGE STANDARD DEVIATION: 30300 3m 4m 43000 57200 56600 51700 53100 61000 62000 48500 12000 Centre Analytical Laboratories, Inc. Page 30 of 102 Centre Study No.:023-063 Table XVII. Summary of PFOS for Week 6 Mallard Membrane Control Samples Sponsor Centre Set Extraction Analysis Analyte E01-0263-20993 EO1-0263-20993* EO 1-0263-20994 E01-0263-20994* EO 1-0263-20995 EO1-0263-20995* EO 1-0263-20975 E01-0263-20975* EO1-0263-20997 E01-0263-20997* 0103764 0103764 0103768 0103768 0103772 0103772 0103692 0103692 0103780 0103780 032701C 3/27/01 3/29/01 NQ 032701C 312710 1 3/29/01 NQ 032701C 312710 1 3/29/01 NQ 032701C 3/27/01 312910 1 NQ 032701C 3/27/01 3/29/01 NQ 032701C 3/27/01 3/29/01 NQ 032701C 3/27/01 312910 1 NQ 032701C 312710 1 312910 1 NQ 032701C 312710 1 3/29/01 NQ 032701C 312710 1 3/29/01 NQ AVERAGE NQ STANDARD DEVIATION NQ Table XVIII. Summary of PFOS for Week 6 Mallard Membrane Samples at 6.2 ppm ai Sponsor Centre Set Extraction Analysis Analyte EO1-0263-20979 EO1-0263-20979* E01-0263-21001 E01-0263-21001* EO1 -0263-20988 E01 -0263-20988 * EOl-0263-20971 EO1-0263-20971* EO1-0263-20989 EO1-0263-20989* 0103708 0103708 0103796 0103796 0103744 0103744 0103676 0103676 0103748 0103748 * Duplicate Injection 032701C 032701C 032701C 032701C 032701C 032701C 032701C 032701C 032701C 032701C 312710 1 312910 1 3/27/01 312910 1 3/27/01 312910 1 312710 1 3/29/01 312710 1 3/29/01 312710 1 3/29/01 3/27/01 3/29/01 3/27/01 3/29/01 3/27/01 312910 1 312710 1 3/29/01 AVERAGE: STANDARD DEVIATION: 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 Analytical Laboratories, Inc. Page 31 of 102 Centre Study No.: 023-063 Table XIX. Summary of PFOS for Week 6 Mallard Membrane Samples at 17.6 ppm ai Sponsor Centre Set Extraction Analysis Analyte E01-0263-20991 E01-0263-20991* EO 1-0263-20990 EO1-0263-20990* EO1-0263-21002 EO1-0263-2 1002* E01-0263-21003 EO1-0263-21003* EO1-0263-21004 EO1-0263-21004* 0103756 0103756 0103752 0103752 0103800 0103800 0103804 0103804 0103808 0 103808 032701CD 312710 1 312910 1 434 032701CD 312710 1 312910 1 43 1 032701CD 3/27/01 312910 1 365 032701CD 312710 1 312910 1 370 032701CD 3/27/01 312910 1 347 032701CD 3/27/01 312910 1 339 032701CD 3/27/01 3/29/01 306 032701CD 3127101 312910 1 307 032701CD 312710 1 312910 1 259 032701CD 312710 1 3129101 26 1 AVERAGE 342 STANDARD DEVIATION 61.2 Table XX. Summary of PFOS for Week 6 Mallard Albumen Control Samples Sponsor Centre Set Extraction Analysis Analyte EO1-0263-20993 EO1-0263-2O993* E01-0263-20994 E01-0263-20994* EO1-0263-20995 E0l-0263-20995* E01-0263-20975 E01-0263-20975* E01-0263-20997 E01-0263-20997* * Duplicate Injection 0103765 0 103765 0103769 0103769 0103773 0103773 0 103693 0103693 0103781 0103781 032701B 3127101 3128-29101 NQ 032701B 3127101 3128-29101 NQ 032701B 3127101 3/28-29/01 NQ 032701B 3127101 3128-29101 NQ 032701B 3127101 3128-29101 NQ 032701B 3127101 3128-29101 NQ 032701B 3/27/01 3/28-29101 NQ 032701B 3127101 3/28-29101 NQ 032701B 3/27/01 3128-29101 NQ 032701B 3/27/01 3/28-29101 NQ AVERAGE NQ STANDARD DEVIATION 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 StudyNo.:023-063 Table XXI. Summary of PFOS for Week 6 Mallard Albumen Samples at 6.2 ppm ai Sponsor Centre Set Extraction Analysis Analyte EO 1-0263-20979 EO 1 -0263-20979* E01-0263-21001 EO1-0263-21001* EO 1-0263-20988 EO1-0263-20988* EO1-0263-20971 E0l-0263-20971* EO1-0263-20989 E01-0263-20989* 0103709 0103709 0103797 0103797 0103745 0103745 0103677 0 103677 0 103749 0 103749 032701B 032701B 032701B 032701B 032701B 032701B 032701B 032701B 032701B 032701B 3127101 3128-29101 3/27/01 3128-29101 3127101 3128-29101 3127101 3128-29101 3127101 3128-29101 3127101 3128-29101 3127101 3128-29101 3/27/01 3128-29101 3127101 3128-29101 3/27/01 3128-29101 AVERAGE: STANDARD DEVIATION: 5.83 6.23 7.92 8.04 6.56 6.26 7.02 6.81 8.72 8.33 7.19 0.987 Table XXII. Summary of PFOS for Week 6 Mallard Albumen Samples at 17.6 ppm ai Sponsor Centre Set Extraction Analysis Analyte E01-0263-20991 E01-0263-20991* E01-0263-20990 ~oi-0263-20990* EO1-0263-21002 E01-0263-21002* E01-0263-21003 EO1-0263-12 W3* EO1-0263-21004 E01-0263-21004* 0103757 0103757 0103753 0103753 0 10380 1 0 10380 1 0103805 0103805 0103809 0103809 * Duplicate Injection LOQ = 10 ppb 032701B 3127101 3128-29101 25.3 032701B 3127101 3128-29101 24.9 032701B 3127101 3128-29101 14.1 032701B 3/27/01 3128-29101 14.7 032701B 3127101 3128-29101 19.7 032701B 3127101 3128-29101 19.4 032701B 3/27/01 3/28-29/10 23.6 032701B 3D7IO 1 3128-29101 24.0 032701B 3127101 3128-29101 19.1 032701B 3127101 3128-29101 17.2 AVERAGE: 20.2 STANDARD DEVIATION: 4.12 Centre Analytical Laboratories, Inc. Page 33 of 102 Centre Study No.: 023463 Table XXIII. Summary of PFOS for Week 6 Mallard Yolk Control Samples Sponsor Centre Set Extraction Analysis Analyte EO1-0263-20993 EO1-0263-20993* EO 1-0263-20994 E01-0263-20994* EO 1-0263-20995 EO 1-0263-20995* EO 1-0263-20975 EO1-0263-20975 * E01-0263-20997 EO1 -0263-20997 * 0103762 0103762 0103766 0103766 0103770 0103770 0103690 0103690 0103778 0103778 032701A 312710 1 3/28/01 NQ 032701A 312710 1 3/28/01 NQ 032701A 312710 1 3/28/01 NQ 032701A 3/27/01 312810 1 NQ 032701A 3/27/01 312810 1 NQ 032701A 312710 1 3/28/01 NQ 032701A 312710 1 3/28/01 NQ 032701A 3/27/01 3/28/01 NQ 032701A 3/27/01 312810 1 NQ 032701A 3/27/01 3/28/01 NQ AVERAGE: NQ STANDARD DEVIATION: NQ Table XXIV. Summary of PFOS for Week 6 Mallard Yolk Samples at 6.2 ppm ai Sponsor Centre Set Extraction Analysis Analyte E01-0263-20979 E01-0263-20979* EO1-0263-21001 E01-0263-21OO1* EO1-0263-20988 EO1-0263-20988* E01-0263-20971 E01-0263-20971* E01-0263-20989 EO1-0263-20989* * Duplicate Injection 0103706 0103706 0103794 0103794 0103742 0103742 0 103674 0103674 0103746 0103746 032701AD 032701AD 032701AD 032701AD 032701AD 032701AD 032701AD 032701AD 032701AD 032701AD 3127101 312910 1 3/27/01 312910 1 312710 1 3/29/0 1 312710 1 312910 1 312710 1 3/29/0 1 312710 1 312910 1 3/27/01 3/29/01 3/27/01 3/29/01 312710 1 3/29/01 3/27/01 312910 1 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 StudyNo.:023-063 Table XXV. Summary of PFOS for Week 6 Mallard Yolk Samples at 17.6 ppm ai Sponsor Centre Set Extraction Analysis Analyte EO1-0263-2099 1 E01-0263-20991* E0l-0263-20990 EO1-0263-20990* E01-0263-21002 E01-0263-21002* E01-0263-21003 EO1-0263-21003* EO1-0263-21004 E01-0263-21004* 0103754 0103754 0103750 0103750 0103798 0103798 0103802 0 103802 0 103806 0103806 032701AD 032701AD 032701AD 032701AD 032701AD 032701AD 032701AD 032701AD 032701AD 032701AD 312710 1 312910 1 3/27/01 312910 1 312710 1 3/29/01 312710 1 3/29/01 3/27/01 3/29/01 3/27/01 312910 1 3/27/01 312910 1 3/27/01 312910 1 3/27/01 312910 1 312710 1 312910 1 AVERAGE: STANDARD DEVIATION: Table XXVI. Summary of PFOS in Quail Membrane Blanks 41900 40700 63000 63500 57500 57300 43100 43600 46600 49800 50700 8870 Sponsor Centre Set Extraction Analysis Analyte na 0003798 Blank A 032901A 3129101 313 110 1 NQ na 0003798 Blank A* 032901A 3/29/0 1 313 1/01 NQ na 0003798 Blank A 033001BR 3/30/01 41210 1 NQ na 0003798 Blank A* 033001BR 3/30/01 4/2/01 NQ AVERAGE: NQ STANDARD DEVIATION: NQ Table XXVII. Surnmary of PFOS in Quail Albumen Blanks Sponsor Centre Set Extraction Analysis Analyte na 0003796 Blank A 032801B 3/28/01 3/30/01 NQ na 0003796 Blank A* 032801B 3/28/01 3/30/01 NQ na 0003796 Blank A 033001A 3/30/01 4/1/01 NQ na 0003796 Blank A* 033001A 3/30/01 4/1/01 NQ AVERAGE: NQ * Duplicate Injection STANDARD DEVIATION: 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 of 102 Centre Study No.:023-063 Table XXVIII. Summary of PFOS in Quail Yolk Blanks Sponsor Centre Set Extraction Analysis Analyte na 0003795 Blank A 032801A 3/28/01 3/30/01 NQ na 0003795 Blank A* 032801A 312810 1 3/30/01 NQ na 0003795 Blank A 032901B 3/29/01 3131-4/1/01 ND na 0003795 Blank A* 032901B 3/29/01 3131-4/1/01 ND AVERAGE: NQ STANDARD DEVIATION: NQ Table XXIX. Summary of PFOS Recoveries in Quail Membrane Sponsor Centre Set Extraction Analysis Amt 9% na 0003798 Spk A 032901A 3/29/01 3131101 10 102 na 0003798 Spk A* 032901A 3/29/01 3131/01 10 104 na 0003798 Spk B 032901A 3/29/01 3131/01 986 88 na 0003798 Spk B* 032901A 3/29/01 3/31/01 986 85 na 0003798 Spk A 033001BR 3/30/01 41210 1 10 114 na 0003798 Spk A* 033001BR 3/30/01 412101 10 116 na 0003798 Spk B 033001B 3/30/01 4/2/01 98 108 na 0003798 Spk B* 033001B 3/30/01 412101 98 101 AVERAGE: 102 STANDARD DEVIATION: 11 RELATIVE STANDARD DEVIATION: 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 of 102 Centre Study No.: 023-063 Table XXX. Summary of PFOS Recoveries in Quail Albumen Sponsor Centre Set Extraction Analysis Amt 9% na 0003796 Spk A 032801B 3/28/01 3/30/0 1 10 82 na 0003796 Spk A* 032801B 3/28/01 3/30/01 10 79 na 0003796 Spk B 032801B 3/28/01 3130101 9963 80 na 0003796 Spk B* 032801B 3/28/01 3/30/01 9963 83 na 0003796 Spk A 033001A 3/30/01 4/1/01 10 82 na 0003796 Spk A* 033001A 3/30/01 4/1/01 10 88 na 0003796 Spk B 033001A 3/30/01 4/1/01 99 93 na 0003796 Spk B* 033001A 3/30/01 4/1/01 99 91 AVERAGE 85 STANDARD DEVIATION: 5 RELATIVE STANDARD DEVIATION: 6 Table XXXI. Summary of PFOS Recoveries in Quail Yolk Sponsor Centre Set Extraction Analysis Amt 9% na 0003795 Spk A 032801A 3/28/01 3/30/01 10 102 na 0003795 Spk A* 032801A 3/28/01 3/30/01 10 104 na 0003795 Spk B 032801A 3/28/01 3/30/01 9623 98 na 0003795 Spk B* 032801A 3/28/01 3/30/01 9623 101 na 0003795 Spk A 032901B 3/29/01 3/31-4/1/01 10 87 na 0003795 Spk A* 032901B 3/29/01 3/31-4/1/01 10 87 na 0003795 Spk B 032901D 3/29/01 4/1-210 1 98834 94 na 0003795 Spk B* 032901D 3/29/01 4/1-2/01 98834 93 AVERAGE: 96 STANDARD DEVIATION: 7 RELATIVE STANDARD DEVIATION 7 * Duplicate Injection LOQ = 10 ppb Centre Analytical Laboratories, Inc. Page 37 of 102 Centre Study No.: 023-063 Table XXXII. Summary of PFOS for Week 1 Quail Membrane Control Samples Sponsor Centre Set Extraction Analysis Analyte EO 1-0259-20506 E01-0259-20506* EO 1-0259-20507 EO1 -0259-20507* EO 1-0259-20520 EO1 -0259-20520* E01-0259-20521 EO1-0259-20521* EO 1-0259-20509 E01-0259-20509* 0101987 0101987 010199 1 010199 1 0102043 0102043 0102047 0102047 010 1999 0101999 032901A 032901A 032901A 032901A 032901A 032901A 032901A 032901A 032901A 032901A 312910 1 313 1/01 3/29/01 313 1/01 3/29/01 3/29/01 3/29/01 3/29/01 312910 1 3/29/0 1 3/29/0 1 312910 1 3/31/01 3/31/01 313 110 1 313 110 1 313 110 1 313 110 1 313 110 1 313 110 1 AVERAGE: STANDARD DEVIATION 5.20** 5.57** NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ Table XXXIII. Summary of PFOS for Week 1 Quail Membrane Samples at 6.2 ppm ai Sponsor Centre Set Extraction Analysis Analyte E01-0259-20526 E01-0259-20526* E01-0259-20514 EO1-0259-20514* EO 1-0259-20527 E01-0259-20527* E01-0259-20515 EO1-0259-205 15* EO 1-0259-20528 EO1-0259-20528* 0102067 0102067 0102019 0102019 0102071 0102071 0102023 0102023 0102075 0 102075 * Duplicate Injection 032901D 032901D 032901A 032901A 032901A 032901A 032901A 032901A 032901D 032901D 3/29/01 4/1-2/0 1 3/29/01 411-2/01 3/29/01 313 110 1 3/29/0 1 313 1/01 312910 1 313 110 1 3/29/01 313 1/01 312910 1 313 110 1 3/29/0 1 313 110 1 312910 1 4Il-UO 1 3/29/01 411-2101 AVERAGE: STANDARD DEVIATION 18800** 17900** 127 129 123 116 172 172 11500** 115oO** 140 25.3 NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) LOQ = 10 ppb ** Possible contamination by yolk or outer shell component of sample since outer shell was not washed prior to separation of internal components. As per Sponsor Representative request these values were not used to calculate the average and standard deviation. For residues recorded as NQ, half of the concentration of the lowest standard (0.5 ppb) was used to calculate the average and standard deviation. Centre Analytical Laboratories, Inc. Page 38 of 102 Centre Study No.: 023-063 Table XXXIV. Summary of PFOS for Week 1 Quail Membrane Samples at 17.6 ppm ai Sponsor Centre Set Extraction Analysis Analyte E01-0259-20529 EO1-0259-20529* EO1-0259-2053 1 E01-0259-20531* EO 1-0259-20532 E01-0259-20532* E01-0259-20518 E01-0259-20518* EO1-0259-20533 EO1-0259-20533* 0102079 0 102079 0102087 0102087 0102091 0102091 0 102035 0102035 0 102095 0102095 032901D 032901D 032901D 032901D 032901D 032901D 032901D 032901D 032901D 032901D 312910 1 41 1-210 1 312910 1 411-2101 3/29/01 411-2JO 1 312910 1 411-2/01 312910 1 411-2/O 1 312910 1 411-2/O 1 3/29/0 1 411-2/O 1 312910 1 411-YO1 312910 1 411-2/01 312910 1 411-YO1 AVERAGE: STANDARD DEVIATION: 20900 20400 1 1000 11100 14300 13900 12500 12600 6880 6990 13100 4730 Table XXXV. Summary of PFOS for Week 1 Quail Albumen Control Samples Sponsor Centre Set Extraction Analysis Analyte EO1-0259-20506 E01-0259-20506* E01-0259-20507 EO1-0259-20507* EO 1-0259-20520 EO1-0259-20520* E01-0259-20521 E01-0259-20521* EO 1-0259-20509 EO1-0259-205O9* 0101988 0101988 0101992 0 101992 0102044 0 102044 0102048 0102048 0102000 0102000 * Duplicate Injection 032801B 032801B 032801B 032801B 032801B 032801B 032801B 032801B 032801B 032801B 312810 1 3130101 NQ 312810 1 3l30lO 1 NQ 312810 1 3130101 NQ 3128101 3130101 NQ 3/28/01 3l30lO 1 NQ 312810 1 3/30/0 1 NQ 3/28/01 3/30/01 NQ 3/28/01 3/30/01 NQ 3/28/01 3/30/01 NQ 3128101 3l30lO 1 NQ AVERAGE NQ STANDARD DEVIATION: NQ NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) LOQ = 10 ppb Centre Analytical Laboratories, Inc. Page 39 of 102 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 EO1-0259-205 14* EO1-0259-20527 EO1-0259-20527* E01-0259-20515 E01-0259-20515* EO 1-0259-20528 E01-0259-20528* Centre ID 0 102068 0102068 0102020 0102020 0102072 0 102072 0 102024 0102024 0102076 0 102076 Set 032801B 032801B 032801B 032801B 032801B 032801B 032801B 032801B 032801B 032801B Extraction Analysis Date Date 3/28/01 3130-31/01 3/28/0 1 3130-3 1/01 3/28/01 3130-31/01 3/28/01 3130-31/01 3/28/01 3130-3 1/01 312810 1 3130-3 1/01 3/28/01 3130-31/01 3/28/01 3130-31/01 3/28/01 3130-31/01 3/28/01 3130-31/01 AVERAGE STANDARD DEVIATION Analyte Found pb) 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 Centre Set Extraction Analysis Analyte EO1-0259-20529 EO1-0259-20529* EO1-0259-20531 EOl-0259-20531* EO 1-0259-20532 EO1-0259-20532* EO1-0259-20518 E01-0259-20518* EO 1-0259-20533 EO1-0259-20533* 0102080 0 102080 0102088 0102088 0102092 0102092 0 102036 0102036 0102096 0102096 * Duplicate Injection LOQ = 10 ppb 032801B 032801B 032801B 032801B 032801B 032801B 032801B 032801B 032801B 032801B 312810 1 3130-3 1/01 17.9 3/28/01 3130-3 1/01 17.3 3/28/01 3130-31/01 9.97 3/28/01 3130-3 1/01 10.5 3/28/01 3130-3 1/01 10.7 312810 1 3130-3 1/01 11.0 3/28/01 3130-31/01 32.3 312810 1 3130-31/01 31.6 312810 1 3130-31/01 9.07 3/28/0 1 3130-31/01 10.1 AVERAGE 16.0 STANDARD DEVIATION: 8.92 Centre Analytical Laboratories, Inc. Page 40 of 102 Centre Study No.: 023-063 Table XXXVIII. Summary of PFOS for Week 1 Quail Yolk Control Samples Sponsor Centre Set Extraction Analysis Analyte EO1-0259-20506 EO1-0259-20506* EO 1-0259-20507 E01-0259-20507* EO 1-0259-20520 EO1-0259-20520* EO1-0259-20521 EO1-0259-20521* E0l-0259-20509 EO1-0259-20509* 0101985 0101985 0101989 0101989 0102041 0102041 0102045 0102045 0 10 1997 0 10 1997 032801A 3/28/01 3/30/01 NQ 032801A 312810 1 3/30/0 1 NQ 032801A 3/28/01 3/30/01 NQ 032801A 3/28/01 3/30/01 NQ 032801A 3/28/01 3/30/01 NQ 032801A 312810 1 3/3W01 NQ 032801A 3/28/01 3/30/01 NQ 032801A 3/28/01 3/30/01 NQ 032801A 3/28/01 3/30/01 NQ 032801A 312810 1 3/30/01 NQ AVERAGE NQ STANDARD DEVIATION NQ Table XXXIX. Summary of PFOS for Week 1 Quail Yolk Samples at 6.2 ppm ai Sponsor Centre Set Extraction Analysis Analyte EO 1-0259-20526 E01-0259-20526* EO1-0259-20514 EO1-0259-205 14* E01-0259-20527 EO1-0259-20527* E01-0259-20515 EO1-0259-20515* EO1-0259-20528 EO1-0259-20528* * Duplicate Injection 0102065 0102065 0102017 0102017 0102069 0 102069 0102021 0102021 0 102073 0102073 032801AD 032801AD 032801AD 032801AD 032801AD 032801AD 032801AD 032801AD 032801AD 032801AD 3/28/01 3/31/01 3/28/0 1 313 110 1 312810 1 313 110 1 312810 1 313 1/01 312810 1 313 110 1 3/28/01 313 1/01 3/28/01 3/31/01 3128101 3131101 3/28/01 313 110 1 3/28/01 313 1/01 AVERAGE STANDARD DEVIATION 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/rd,)) LOQ = 10 ppb Centre Analytical Laboratories, Inc. Page 41 of 102 Centre Study No.: 023-063 Table XL. Summary of PFOS for Week 1,QuailYolk Samples at 17.6 ppm ai Sponsor Centre Set Extraction Analysis Analyte EO1-0259-20529 EO 1-0259-20529* EO1-0259-20531 EO1-0259-20531* EO1-0259-20532 EO1-0259-20532* E01-0259-20518 E01-0259-20518* EO 1-0259-20533 EO1-0259-20533* 0102077 0102077 0102085 0102085 0102089 0 102089 0102033 0102033 0102093 0 102093 032801AD 032801AD 032801AD 032801AD 032801AD 032801AD 032801AD 032801AD 032801AD 032801AD 312810 1 313 110 1 3/28/01 313 110 1 3/28/01 3/31/01 3/28/01 3/31/01 3/28/01 313 110 1 3/28/01 313 110 1 3/28/01 313 1/01 3/28/01 313 1/01 3/28/01 313 1/01 3/28/01 313 1/01 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 E01-0259-20735 E01 -0259-20735 * EO1-0259-20736 EO1-0259-20736* E01-0259-20737 E01-0259-20737* EO 1-0259-20720 E01-0259-20720* Eo1-0259-20738 EO1 -0259-20738* * Duplicate Iniection 0102903 0 102903 0102907 0102907 0102911 0102911 0102843 0102843 0102915 0102915 033001B 033001B 033001B 033001B 033001B 033001B 033001B 033001B 033001B 033001B 3/30/01 4WO 1 3/30/01 3/30/01 4/UO 1 4/UO 1 3/30/01 41210 1 3/30/0 1 4/UO 1 3/30/01 4/UO 1 3/30/01 4lUO 1 3/30/01 3/30/01 41210 1 4lZO 1 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 Representativerequest 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 E01 -0259-20742 EO1-0259-20742* EO1-0259-20743 EO1-0259-20743* EO 1-0259-20744 EO1-0259-20744* E01-0259-20745 EO1-0259-20745* EO 1-0259-20746 EO1-0259-20746* 0102931 0102931 0102935 0102935 0102939 0 102939 0102943 0 102943 0102947 0102947 033001B 3l30lO 1 4nIO 1 120 033001B 3l30lO 1 4/2/0 1 119 033001B 3130101 4/2/01 213 033001B 3l30lO 1 41210 1 208 033001B 3l30lO 1 4/2/0 1 123 033001B 3l30lO 1 41210 1 116 033001B 3/30/01 41210 1 109 033001B 3/30/01 41210 1 116 033001B 3/30/01 41210 1 134 033001B 3l30lO 1 41210 1 134 AVERAGE: 139 STANDARD DEVIATION: 38.4 Table XLIII. Summary of PFOS for Week 6 Quail Membrane Samples at 17.6 ppm ai Sponsor ID E01-0259-20747 EO1-0259-20747* EO 1-0259-20748 EO1-O259-20748* EO1-0259-20749 EO1-0259-20749* E01-0259-20750 E01-0259-20750* E01-0259-20751 E0l-0259-20751* * Duplicate Injection LOQ = 10 ppb Centre ID 0102951 0102951 0 102955 0 102955 0102959 0102959 0102963 0102963 0 102967 0102967 set , Number 033001BD 033001BD 033001BD 033001BD 033001BD 033001BD 033001B 033001B 033001B 033001B Extraction Analysis Date 3130l01 3130101 3l30lO 1 3130101 41210 1 4/2/01 4WO 1 41210 1 3130101 3l30lO 1 41210 1 4/2/01 3130101 3/30/01 41210 1 4/2/01 3l30lO 1 3l30lO 1 41210 1 41210 1 AVERAGE: STANDARD DEVIATION: Analyte 405 389 318 318 394 400 262 250 186 185 311 86.6 Centre Analytical Laboratories, Inc. Page 43 of 102 Centre Study No.: 023-063 Table XLIV. Summary of PFOS for Week 6 Quail Albumen Control Samples Sponsor Centre Set Extraction Analysis Analyte EO1-0259-20735 E01-0259-20735* EO1-0259-20736 EO1-0259-20736* E01-0259-20737 EO1-0259-20737* E01-0259-20720 EO1-0259-20720* EO 1-0259-20738 EO1 -0259-20738* 0102904 0102904 0102908 0 102908 0102912 0102912 0102844 0102844 0102916 01029 16 033001A 3/30/01 4/1/01 NQ 033001A 3/30/01 4/1/01 NQ 033001A 3/30/01 4/1/01 NQ 033001A 3/30/01 4/1/01 NQ 033001A 3/30/0 1 4/1/01 NQ 033001A 3/30/0 1 4/1/01 NQ 033001A 3/30/01 4/1/01 NQ 033001A 3/30/01 4/1/01 NQ 033001A 3/30/01 4/1/01 NQ 033001A 3/30/01 4/1/01 NQ AVERAGE NQ STANDARD DEVIATION NQ Table XLV. Summary of PFOS for Week 6 Quail Albumen Samples at 6.2 ppm ai Sponsor Centre Set Extraction Analysis Analyte EO 1-0259-20742 EO1-0259-20742* EO 1-0259-20743 EO1-0259-20743* EO 1-0259-20744 E01-0259-20744* E O 1-0259-20745 E01-0259-20745* EO1 -0259-20746 E01-0259-20746* * Duplicate Injection 0102932 0102932 0102936 0102936 0102940 0102940 0102944 0102944 0102948 0102948 033001A 3/30/01 4/1/01 5.34 033001A 3/30/01 4/1/01 5.93 033001A 3/30/01 4/1/01 6.19 033001A 3/30/0 1 4/1/01 6.91 033001A 3/30/0 1 4/1/01 6.82 033001A 3/30/0 1 4/1/01 6.82 033001A 3/30/01 4/1/01 5.26 033001A 3/30/01 4/1/01 5.15 033001A 3/30/01 4/1/01 3.56 033001A 3/30/0 1 4/1/01 4.34 AVERAGE: 5.63 STANDARD DEVIATION: 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 Centre Set Extraction Analysis Analyte EO 1-0259-20747 E01-0259-20747* E01-0259-20748 EO1-0259-20748* E0l-0259-20749 E01-0259-20749* EO1-0259-20750 EO1-0259-20750* E01 -0259-2075 1 E01-0259-20751* 0 102952 0 102952 0 102956 0 102956 0102960 0 102960 0102964 0 102964 0102968 0102968 033001A 3/30/01 4/1/01 9.81 033001A 3/30/01 4/1/01 9.84 033001A 3/30/01 4/1/01 13.2 033001A 3/30/01 4/1/01 13.1 033001A 3/30/01 4/1/01 14.6 033001A 3/30/01 4/1/01 14.2 033001A 3/30/01 4/1/01 16.3 033001A 3/30/01 4/1/01 16.7 033001A 3/30/01 4/1/01 15.6 033001A 3/30/01 4/1/01 15.4 AVERAGE: 13.9 STANDARD DEVIATION: 2.44 Table XLVII. Summary of PFOS for Week 6 Quail Yolk Control Samples Sponsor Centre Set Extraction Analysis Analyte EO1 -0259-20735 EO1-0259-20735* E01-0259-20736 EO1-0259-20736* EO 1-0259-20737 EO1-0259-20737* EO 1-0259-20720 EO1-0259-20720* E01 -0259-20738 E01-0259-20738* * Duplicate Injection 0102901 0102901 0 102905 0102905 0102909 0102909 0102841 0102841 0102913 01029 13 032901B 032901B 032901B 032901B 032901B 032901B 032901B 032901B 032901B 032901B 312910 1 3/29/01 3/29/01 3/29/01 3/29/01 313141101 3131-4/1/01 3131-4/1/01 3131-4/1/01 3131-4/1/01 3/29/01 3131-4/1/01 3/29/01 3131-411/01 3/29/01 3131-4/1/01 3/29/01 313 1-4/1/01 3/29/01 3131-4/1/01 AVERAGE STANDARD DEVIATION: NQ NQ NQ NQ 51.5** 51.1** NQ NQ ND ND NQ NQ NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) N D = 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 Centre Study No.: 023-063 Table XLVIII. Summary of PFOS for Week 6 Quail Yolk Samples at 6.2 ppm ai Sponsor Centre Set Extraction Analysis Analyte EO 1-0259-20742 E01 -0259-20742* EO 1-0259-20743 EO1-0259-20743 * EO 1-0259-20744 EO1 -O259-20744* E01-0259-20745 E01-0259-20745* EO1-0259-20746 EO1-0259-20746* 0102929 0102929 0102933 0102933 0102937 0102937 0102941 0102941 0102945 0102945 032901D 032901D 032901D 032901D 032901D 032901D 032901D 032901D 032901D 032901D 3/29/01 4/1-20 1 3/29/01 411-201 3/29/01 411-201 3/29/01 411-201 3/29/01 411 -210 1 3/29/01 411-2101 3/29/01 411-201 312910 1 411-2101 3/29/01 411-2101 3/29/01 411-2101 AVERAGE STANDARD DEVIATION: 387005 40500 37 100) 36100 23600, 23600' 4m\ 406w 24600, 25 8oo.i' 33100 7610 Table IL. Summary of PFOS for Week 6 Quail Yolk Samples at 17.6 ppm ai Sponsor Centre Set Extraction Analysis Analyte EO1-0259-20747 EO1-0259-20747* E01-0259-20748 E01-0259-20748* E01 -0259-20749 E01-0259-20749* EO1-0259-20750 EO1-0259-20750* EO1-0259-20751 E01-0259-20751* * Duplicate Injection 0102949 0102949 0 102953 0 102953 0102957 0 102957 0 10296 1 0102961 0102965 0102965 032901D 032901D 032901D 032901D 032901D 032901D 032901D 032901D 032901D 032901D 312910 1 411- a 0 1 312910 1 411-2101 312910 1 411-2/01 3/29/01 411-YO1 3/29/01 411-201 3/29/01 411-201 3/29/01 4/1-2/10 312910 1 411-YO1 3/29/01 411- 2 01 312910 1 411-201 AVERAGE STANDARD DEVIATION 53900, 54400' 65500 648d :E3 59400, 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 of 102 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 EO1-0263-20825 E01-0263-20846 EO 1-0263-20832 E01-0263-20847 E01-0263-20848 EO 1-0263-20856 E01-0263-20850 E01-0263-20857 E01-0263-20851 EO1-0263-20858 E01-0263-20993 E01-0263-20994 EO 1-0263-20995 EO 1-0263-20975 E01-0263-20997 E01-0263-20979 E01-0263-21001 EO 1-0263-20988 E01-0263-20971 EO 1-0263-20989 EO1-0263-20991 E01-0263-20990 E01-0263-21002 E01-0263-21003 E01-0263-21004 Centre ID 0103200 0103204 0103208 0103160 0103164 0103092 0103176 0103120 0103180 0103184 0103216 0103192 0103220 0103196 0103224 0103764 0103768 0 103772 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 .O 85.0 85.0 75.0 71.4 80.0 65.O 80.0 75.0 65.O 75.0 90.0 80.0 75.0 70.0 70.0 75.0 65 .O Centre Analytical Laboratories, Inc. Page 47 of 102 Centre Study No.: 023-063 Table LI. Summary of Moisture Determinationsfor Mallard Albumen Samples Sponsor Centre Average E01-0263-20852 0 103201 90.0 EO1 -0263-20853 0103205 88.0 EO 1-0263-20854 0103209 87.0 EO 1-0263-20842 0103161 85 .O E01-0263-20843 0103165 87.0 E01-0263-20825 0103093 82.0 EO1-0263-20846 0103 177 84.0 E01-0263-20832 0103121 86.0 EO 1-0263-20847 0103181 89.0 E01-0263-20848 0103185 88.0 EO1-0263-20856 0 1032 17 85.0 E01-0263-20850 0103193 85.0 EO1-0263-20857 0103221 84.0 E01-0263-20851 0103197 83.0 EO 1-0263-20858 0103225 85.0 EO 1-0263-20993 0103765 88.0 E01-0263-20994 0103769 85.0 E01-0263-20995 0103773 85.0 E01-0263-20975 0103693 84.0 EO1-0263-20997 0103781 90.0 EO1-0263-20979 0103709 88.0 EO1-0263-21001 0103797 84.0 E01-0263-20988 0103745 87.0 E01-0263-20971 0103677 83.0 EO 1-0263-20989 0 103749 87.0 EO 1-0263-20991 0103757 84.0 E01-0263-20990 0103753 86.0 EO1-0263-2 1002 0 103801 83.0 EO1-0263-2 1003 0103805 82.0 E01-0263-21004 0103809 87.0 Centre Analytical Laboratories, Inc. Page 48 of 102 Centre Study No.: 023-063 Table LII. Summary of Moisture Determinationsfor Mallard Yolk Samples Sponsor ID E01-0263-20852 EO1-0263-20853 E01-0263-20854 E01-0263-20842 E01-0263-20843 EO1-0263-20825 E01-0263-20846 EO1-0263-20832 EO 1-0263-20847 EO 1-0263-20848 EO1-0263-20856 EO1-0263-20850 EO 1-0263-20857 EO1-0263-20851 EO 1-0263-20858 EO 1-0263-20993 EO 1-0263-20994 E01-0263-20995 EO1-0263-20975 EO 1-0263-20997 EO1-0263-20979 EO1-0263-21001 EO1-0263-20988 EO1-0263-20971 EO 1-0263-20989 EO1-0263-20991 E01-0263-20990 EO1-0263-21002 E01-0263-21003 E01-0263-21004 Centre ID 0103198 0 103202 0103206 0103158 0103162 0103090 0103174 0103118 0103178 0103182 0103214 0103190 0103218 0103194 0103222 0103762 0103766 0103770 0 103690 0103778 0103706 0103794 0103742 0103674 0103746 0103754 0103750 0103798 0103802 0103806 Average % Moisture 44.0 45 .O 48.0 45 .O 43.0 54.0 49.0 43.0 43.0 48.0 45.0 50.0 49.0 48.0 45 .O 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 of 102 Centre Study No.: 023-063 Table LIII. Summary of Moisture Determinations for Quail Membrane Samples Sponsor Centre Average E01 -0259-20507 0101991 45 .O E01-0259-20509 010 1999 65.O E01-0259-20526 0102067 65.0 EO 1-0259-20738 0102915 60.0 EO 1-0259-20742 0102931 60.0 EO 1-0259-20745 0102943 68.3 EO 1-0259-20748 0 102955 70.0 EO1-0259-20750 0102963 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 Determinationsfor Quail Albumen Samples - Sponsor ID E01-0259-20506 EO 1-0259-20507 EO 1-0259-20520 EO1-0259-2052 1 EO 1-0259-20509 EO 1-0259-20526 EO1-0259-205 14 EO 1-0259-20527 E01-0259-20515 E01-0259-20528 EO 1-0259-20529 E01-0259-20531 EO 1-0259-20532 EO1-0259-20518 EO 1-0259-20533 EO 1-0259-20735 EO 1-0259-20736 EO 1-0259-20737 EO 1-0259-20720 EO 1-0259-20738 EO 1-0259-20742 E01-0259-20743 E01-0259-20744 EO 1-0259-20745 EO 1-0259-20746 EO1-0259-20747 EO 1-0259-20748 EO 1-0259-20749 E01-0259-20750 EOl-0259-20751 Centre ID 0101988 0 101992 0102044 0102048 0102Ooo 0102068 0102020 0102072 0102024 0102076 0102080 0102088 0102092 0 102036 0102096 0102904 0102908 0102912 0102844 01029 16 0 102932 0102936 0102940 0102944 0 102948 0102952 0102956 0102960 0102964 0102968 Average 9% Moisture 87.0 79.0 89.0 88.0 85.2 87.0 88.0 86.0 89.0 87.0 89.0 88.0 88.0 87.0 87.0 88.0 86.0 87.0 87.0 87.0 86.0 82.0 84.0 87.0 86.0 89.0 86.0 85.0 87.0 86.0 - 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 EO1-0259-20506 EO1-0259-20507 EO1-0259-20520 E01-0259-20521 EO 1-0259-20509 EO 1-0259-20526 E01-0259-20514 EO 1-0259-20527 E01-0259-20515 EO1-0259-20528 EO 1-0259-20529 E01-0259-20531 EO 1-0259-20532 EO1-0259-20518 EO 1-0259-20533 EO1 -0259-20735 E01-0259-20736 E01-0259-20737 E0l-0259-20720 EO 1-0259-20738 EO1 -0259-20742 E01-0259-20743 E01-0259-20744 E01-0259-20745 E01-0259-20746 E01 -0259-20747 EO 1-0259-20748 E01-0259-20749 E01-0259-20750 EO1-0259-2075 1 Centre ID 0101985 0 101999 0102041 0102045 0101997 0102065 01020 17 0 102069 0102021 0102073 0 102077 0102085 0102089 0102033 0102093 010290 1 0102905 0102909 0102841 0 1029 13 0102929 0 102933 0102937 010294 1 0102945 0102949 0102953 0 102957 0102961 0 102965 Average % Moisture 48.0 43.0 46.0 45.0 45.0 49.0 49.0 47.0 54.0 47.0 47.0 41.0 49.0 50.0 47.0 47.0 46.0 47 .O 41.0 43.0 44.0 43.0 45.0 53.0 46.0 47.0 46.0 45.0 45 .O 43.0 Centre AnalyticalLaboratories,Inc. Page 52 of 102 Centre Study No.:023-063 FIGURES Centre Analytical Laboratories, Inc. Page 53 of 102 . . . 0 . 0 ' . I . , . - .I . 1 . I ~ 1 ~ 1 ~ I ~ I . , ~ , ~ , . , . , . , . , _ ) . , _ , . , _ , 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 2.4 2.6 2.8 3.0 3.2 3.4 3.6 3.8 4.0 4.2 4.4 4.6 4.8 5.0 %2 _ _ ---.-_ Concentration, n g h L Centre StudyNo.:023-063 Figure 2. Chromatogram Representing a 0.1 ng/mL standard for PFOS 1.. 1 S.06 asao a 30.0 a.5 4.41 ria 0.10 0.01 s.00 4.00 1-00 mo Centre Analytical Laboratories, Inc. Page 55 of 102 Centre Study No.:023-063 Figure 3. Chromatogram Representing a 1.0 ng/mL standard for PFOS 0 CM 3s 2 0 - 0 45 5.0 u LO 65 7.0 75-I Centre Analytical Laboratories, Inc. Page 56 of 102 Centre Study No.: 023-063 Figure 4. Chromatogram Representing a Reagent Blank for PFOS (Centre ID Reagent Blank A, Set: 0326018) .- . COILC8UCZAClOUr k q . Mcer Mq. T 1 r . r 0.000 01126101 04:52:58 ?tl 18. 1 5.06 15.30 2 30.0 S.C 4.43 .in 0.10 0.01 9.00 U..i rn.lQhCt Scafc t i v r .nd t l r r m.1 4.4s 490.3 l.5k.001 4.37 4.58 sin COODU .in ria 0' l.o 1.s io is io is i o 4 3 co . 5s i o 6 5 7.0 7.5 l!ens.ml I Centre Analytical Laboratories,Inc. Page 57 of 102 Centre Study No.:023-063 Figure 5. Chromatogram Representing Control Mallard Membrane for PFOS (Centre ID: 0003802 Blank A, Set: 032601C) Centre Analytical Laboratories, Inc. Page 58 of 102 Centre Study No.: 023-063 Figure 6. Chromatogram Representing Control Mallard Membrane Fortified with 10 ppb of PFOS, (Centre ID:0003802Spk A Dup. Inj., Set: 032601C) Centre Analytical Laboratories, Inc. Page 59 of 102 I Centre Study No.: 023-063 Figure 7. Chromatogram Representing Mallard Membrane Sample from Week 1 at 6.2 ppm ai (E01-0263-20825) (CentreID:0103092, Set: 032601C) Centre Analytical Laboratories, Inc. Page 60 of 102 ~~~~ ____ Centre Study No.:023-063 APPENDIX A Study Protocol OOP-023-063 (Centre Study No. 023-063) and Amendments and Deviations Centre Analytical Laboratories, Inc. . .. Page 61 of 102 ._ __ . .-._ . . Centre Study Not023463 Cenae Protocol No. OOP-023-063 A " X l J C A L PHASE PROTOCOL SPONSOR 3M Environmental Technology and Safety Services Building 2-3E-09 PO Box 33331 St.Paul, IvN 55133-3331 DATA REQUIREl"B Analytical Method Requirements PERFORMING LABORATORY Centre Analytical Laboratories,Inc. (Centre> 3048 Research Drive State College, PA 16801 Phone 814-231-8032 PROTOCOL IDENTIFICATIONNUMBER ooP-023-063 Centre Analytical Laboratories, Inc. .. . P a p 1of 38 Page 62of 102 .. . ... . .. .. Centre Study No.: 023-063 Centn Protocol No. OOP023-063 Title: EXTRACTION OFPOTASSIUMP ~ U 0 R ~ " E S U L F O N A TFREOM EGG MEMBRANE,YOLK AND ALBUMENFOR ANALYSIS USINGHPLCELE~~TROSPRAY/MSAPSESCIROMERY TABLEOF C 0 " T S - TABLE OF CO-S Page _...................................................--- --................. 2 I 1. PT.JRPOS..E...................................... -..............,.........-.... .-e ................ 3 2. REFERENCEMATERIAL............ ".-..........-..----. -.*.-............. 3 3. SPONSOR ........................................................................... ......................".......4 4. m G FACILJTY.............................................. I----.-.- .................................. 4 5. PERFORMINGLABORATORY..........-.-..............------................................... 4 6. PROPOSEDE X P ~ A TILME-FRAME----------................ 4 7. JUSTIFICATION FOR "HJiSELECTIONOFTHE"EXf SYSTEM................... 4 8. SAMPLEPROCESSING,STORAGEAND JDE"JlWA'l''ION .............. 5 9. ANALyTicALMETHOD .............-........... -.-.--_. ................................. 5 10. m m aDESIGN.............................-.......-....--.- ................. 5 11. PROTOCOLAMENDMENTS AND DEVIATIO"IT-----.-................. 6 12. ~ C O ......~.........S............................................... -......--...--.---.-.............................6 13. Q U M ASSURANCE._..........-........-...........--.-.-.--..-.-...-...-............................ 7 14. DATA AND REPORT........................................................ ............................. 7 15. A R m STATEMENT.......................... "-.... ................ 8 16. REFERENCES....................................... . . . . . . . . . . . . . . . . . . . . . . . . . ".................. 8 17. PROTOCOLAppROvAL ...............-................................. .....................................9 AppENDn: MAL~TICALMI"OD ..........-....................-..-.--...................................10 centreM y t i c a l Labomories,Inc. Centre Analytical Laboratories, Inc. -Page 2 of 38 Page 63 of 102 Centre ProtocolNo.OOP-023-063 me analytical phase will be conducted under EPA TSCA Good Laboratory Practice standards40CFR Pat 792 (1). PFOS TCR00017-46 I 97.9 I ~ -~ 08/31/01 0 0 Centre Analytical Laboratories. In& Centre Analytical Laboratories,Inc. Page 3 of 38 Page 64 of 102 ~~ .-.. ...................... - .... .... ._....._. . . ............ . . . . . . . . "CentreStudy No.: 023-063 HAZARD INFORMATION Centre Protocol No. CQP-023-063 3 M EnvironmentalTechnologyand SafetyServices Building 2-3E49 Po Box 33331 St. Paul, MN 55133-3331 Sponsor Representative: RochelIe Robideau TELEPHONE: 651-778-7065 FAX NUMBER: 651-7786176 4. TESTINGFACILITY WiIdlife International,Ltd. 8598 Commerce Drive Easton, MD '21601 STUDY DIRECTOR Sean GaUagher TELEPHONE 410-822-8600 FAX NUMBER:410-822-0632 5. PERFORMING LABORATORY Cmtn Analytical Laboratories, Inc. (Centre) 3048 Research Drive State College, PA 16801 PRINCIPAL INVESTIGATOR EmilyR.Stauffcr, Ccnm TELEPHONE: 814-231-8032 FAXNUMBER: 814-231-1580 6. PROPOSEDEXF'ERlMEMTAL " I h 4 E - m Analytical Start Date Analytical Tnmination Date Report Issued Mar. 19,2001 Mar. 31,2001 April 1,2001 Centre Analyticd Laboratories. hc. Centre Analytical Laboratories, Inc. ........... ,. Page 4 of 38 . . . . Page 65 of 102 Centre Study No.:023-063 Centre Protocol No. OOP-023-063 9. ANALYTICALMETHOD 10. EXPERIMENTALDESIGN Centre Analytical hthatoriesk Centre Analytical Laboratories, Inc. Page 5 of 38 Page 66 of 102 .. . . . .... CentreStudy No.:023-063 Centre Protocol No. OOP423-063 11. PROTOCOLAMENDMENTSAND DEVIATIONS 2. p r o t ~dleviations: Protocol deviations. which are one time and unplanned deviationsfromthe proto~~shl,all be docu~ntedin the study records, noting the n e of the deviation, potential effect or impact on the study, and c o d v e action if required. Protocol d e v h h ~ signedand dated by the Investigator, Study Director. and Sponsor Representative and reviewcdby QAU. 12. RECORDS Records to maintained include, but are not limited the following (as appropriate): 1.Sampleacking shect(s1 2. Samplereceiptrecords,storage history. and chains of custody 3. &tMy ;md p n p d o n of stvldards (Stock, foItification, calibration) Centre Analytical L3bOntorics. Inc. Centre Analytical Laboratories, Inc. Page 6 of 38 Page 67 of 102 . . .. . . .. . . ,. -... . _.__..... ... . . - . .- I . . .. .. . .. . ' Centre Study No.:.023-063 . . .. . . I- Centre Rotocoi No. OOP423-063 4. Descriptionof any modificationsto the method 5. Instrumentm sheets, bench-sheetsor logs 6. Analytical data tables 7. All chromatographic and instrumental conditions 8. Sample extraction and analysis dates 9. A completelisting of studypersonnel,signaturesand initials 10.Chronologicalpresentationof all studycorrespondence 11.Any other data neceSSary for the reconstructionof the study All chromatograms will contain the following: Each data set will contain information on temperatuns, flow rates, column param#ers,g a s , instxument panmeten, and instrument type, etc. n e cmb study number will also be recorded on the first chromatogram of each daily analytical m. 13. QUALITYASSURANCE Cenb QA Unit will review the protoCOl, audit the study conduct and the study documentation (including raw data and final Rport) to ascertain that all GLp st;m&& an met. Centre QA Unit will inspect the study at intervals adequate to assure compliance to GLP's, and will npoa the findings of the audits to the principal Investigator, Centre Management, and the Study Director. 14. DATA AND REPORT 1. All raw data and the original signed ptoCO1 will be maintained in the study file. This data includes the p r o ~ o lamendments, protocol deviations, labomtory notebooks. analficd Standard SOlUtiOn preparation, sample chain of custody sheets. sample work sheeu, Chromtognms, calibration curves, and any other appropriatedata generated. Raw data not used will be s t o d in the studyfile.Thew o n for exclusionwill be documented. 2. A nport will be issued by Centre, approved and signedby the Study k t o r and the Sponsor Rcpnsentativc and sufficient for submission to EpA. m e npoa contents should include, but not limited to: 1. Objectivesand procedures stated in the protocol 2 Analytical and mtistical methods used Centre AnalyticalLaboratories,Inc. h g e 7 O f 38 Page 68 of 102 -- Centre Study No.: 023-063 C e n t P~r~wccol No.OOP-023-063 3. Reference materialsidentifiedby name, lot, purity, and other characteristics 4. Name of performing laboratory a d analytical SM and -nation dates 5. Statement prepared and signed by the quality assurance unit 6. Tables containingall applicable data 7. All chromatographic and instrumental conditions 8. A complete listing of Centre study personnel REPORT DISTRIBUTION: Original: Study Director copy: CentreArChiVes Document archives: Upon completion Of the study, the study ncords, protocol and amendmenu, and the final report and mdments shall be retained in the 3M document archives. If it is ncccSSary to Substitute a COPY for an originid ncord, it will be certified as an exact copy. Centre will retain facility-rclated originals and a copy of the final npon and raw data p a c k e . 16. REFERENCES 1 U.S. Environmental Protection Agency. Toxic Substances Control Act (TSCA). Good Labomtory Practice Standards. Final Rule, 40 CFR Part 792. Centre M y t i c a l ~ r i u o r i c sI,nc. Centre Analytical Laboratories, Inc. Page 8 of 38 Page 69 of 102 Centre Study No.: 023-063 . Cen- Protocol No. OOP423-063 17. PROTOCOL APPROVAL This protocol was audited by the Quality Assurance Unit of Centre Analyticai Laboratories, k Quality Asnvance Reviewer, Centre U A b - : @LL(!/L Naomi Lovallo, W . Quality Assurance Auditor 'PrincipalInvestigator, Centre n Facility Management, Centre Study Director, Wildlife Intern?tional, Ltd. Date SenioGrdBai-o ogkt Sponsor, 3M F . CenmAnalyticalmais,Inc. Centre Analytical Laboratories,Inc. Page 9 of 38 Page 70 of 102 ... Centre Study No.:023-063 1 @PENDM: ANALYTICAI. METHOD A "Dewnation of Pedluorooctanesulfonate in Egg Membrane, Albumen, and i Yoik by Lc/Ms/MS". Centre method number OOM-023415 .. - ......... ...:'.. . . Centre Mytical Laboratories,Inc. Page 10of 38 .Page 71 of 102 .... ... CentreStudy No.: 023463 .. . .. . . .. .' . . . . . . rr,.- . . . . . *- 1 . . . - . . 5: *.. .- .. .. - TOTALNUMBEROFPAGES . ' -13 * .. ... .. ..? ' I .! . - -.... . _. ... -* ........... . . .* . .. . . . . . . .- .. ." - .. . -. . I beAne alyiicj! :. GK&i-..s;Fi.-. - . . . ... ..-.--. . . . . . . ..-.-._ .. . ._ . Centre Analyticai Latmrattries,~ n c . .. . . .. , . .- I 11.of 38 .. * Page 72 of 102 ... _-.......... ...... ~ Centre Study No.: 023-063 .. * . . * .. . .. Centre Analytical Labbratories, IUC. .. . *. . *. ... _. I ha2 . - . ......2. . . .. -.-. .- * '-..Page E of 38 Page 73 of 102 Centre Study No.: 023-063 . -.. j 1" I I .. .....--.--................ . . .--I -.":.&- - - .....:.... ..... . .-. ... - - 7 ---> - ." ' '.. i I .. .. .: . TABLEOFCONTENTS PAGF kR0Y.U .. .T&.EOFCON"TS . ' 9OFTABUS UsTOFrlGURFs .* .-.. 1. . -. . . -2 . . '-3 .- ; -* I 4 ... -. *. ... .. .-. 6. S-" . . i. ' . -. * . .. I ... 1 .. - - - ._. . . '-' ......... . . e.- ..2.. .. .. ..... ..--:-......... ...... Centre Analytical Labor;;tories;Inc. ' 162 ......................... ...... .... . . . . . Centre SWdy No.:023-063 ...... ........ 1 : . . . . . . . . . . . . . . .. . .. . . . . . . . . . . . . . . . . - . . ... : &xic pnrtocol No. OOP-023p63 ..___......... ...-..-...-". . : . '. ? .. 1 I-. .. .---.=L-- .. .- '.' .cult.reMakd No: OO&I.(J~.(J1j I .. ... - . . .- . . . .- ' .. - .. . .. . . - 1 - . - .e .. .. ... * . . . _. .. ...- . . . .* .. - . * Page 4 ... ... Centre Study No.: 023-063 . . . .... ........ . .-. . . -.. - _ .. i .... .. .. :-. . . --__ ... . ... ........ . ....... . . . . .*: Ceme protocblNo.'OoP-023-0,53 .......... -" -- ._ _. ... f.Jt"-' .. L. _.......... .---A::::-:::.. ... '. . !.. .- .. .. ... . .. . . . .... .' a . .. .. .. -. .. __ . . . ! d' ........... -.--. .. . e. .......-- ... .- ...... . ... .. . . .. .. . .. - .. . . Pas5 - . . - . , .... . E .. 0 . .. Centre Analytical Laboratories, Inc. Page 76 of 102 . .' . .. .. .. .. I t .---. .---- .- .. . . ! : !. .: I .- .. .. -. Centre Study No.:023. -. 063 .. . ... . centre A n a l y t i c a L L e s , ~ n c . Centre Study No.:023-063 .. ..-_.-..._ ____. .. . ... 1 2. e . . .* P - . I e&O-K+ U I . 0 .. , .. i ::. .. .. .. *-:3. .. - .. . .. .. . : -3:. .. . . - . cHag~:&rnPLlEs. - !.I: C k g A u * - . :::. . - - . .* .. . . - ..... -:. - . . . .- -. . .- .. .. - .: - -. *.. *z: - .. . ..-.. .. I .. 3 .. .......... - .._ .-.- .- - ..-.. . .. ! . - .. ....- . .. . .-. Page 17 of 38 .. . . Page 78 of 102 . . ._..;....._.-.._._ . .. .. . . .. .... ... .. ... .Centre Study No.:023-063 .. I .. .. .. I .. .. .. .. .. .. .. .. . / .' - .. .. . .- . ....- .. ..: * - Page 79 .of 102 Centre Study No.:023-063 .. -- . . . . . ............. .__.-............ .-- . . -..... .. . *: centre Rotocblr;&. jp.JJ&a3 . "' .. a. I . .... . i. .. .. .. .. .. .. .. .. .. .. -. . . -. centre -;tical .. Labotatoies,~ n c . . .. : Page 80 of 102 .. I t I' I Centre Study .. No..:. 0. 2. 3. -063 .- . . .-- .'. . CentreAnalytical Laboratories,Inc. .. ' Page 81 of 102 .. ' .. .. : ' ... . . - . .. . .. .. . Centre Study No.:023-063 . ------ - .. ). .. . .. i .... . . ?L I .. . . :. *:. Centre Study No.:023-063 . ' .. .. * e .. - ... .. i . .. . . .. .. .. . . . . .... .. . . . . . . . "___..... .. . . ^ . ;. . . .- - ..- . . a . Page 83 of 102 .... Centre Study No.: 023-063 . . . . . . --. .- i .. . ' .... --. -___-_.* . ... .. ._.__... ... I-..- . . . . . - . I Page 84 of 102 .' .. Centre Study No.:023-063 .. 1- .- , - ' ..- .. .. -.. . . ..i .. .. - f 1. . . .. .. . .. -. .! . . ..- ... . ...-... .. .. *. . Page 85 of-102 '1. .! . .... .. ' ! :. . . i Centre Study No.:023-063 . - . . .- .............-...-.. .... . .. . . . . .5 . ' . .......................... -i-....&--.L-, - - .. . . . ..''a .. NQ.00p423463 . . . - * . e*-.-.-,, .* .. .. .. .. .. . . . ., - . -- . . ... .8' .. a. .. , .. .. .. .. .*. . . . *. . .. '. .. .- , :. ' . . . .. _-_.. centre Analytical Laboratories, Inc. " . ......... ._ I %e 25 of 38 Page 86 of.102 ...... .. ......... .. , . .- Centre StudyNo.:023-063 . .- - . . ............ - . . --..-- -_._ . .~ . * I . - . . - . . . . . . . e..--........ .' . -. ...*..-. .- :* .---.. ._. . . ............-...... .. ...I . .A. Centre Study No.:023-063 - ... . -.. - . . . . .---. . . . ...-. . .. ----.---- .. .. I .. * Ci&-k-lN6:.mpm.?ooP..3d6j . . - . . . . . . . . . . L 2' . * . s- ' . .- . . . . .. . . -. .. .. . I e. . . . . * . . . .:.. .. .. I' , e . .. . . .. e. -. . .-. - !... .-. . .. . - .. . . . . . .. ... .. Lauc hi-& :DltOlS .. .. - .- k 17 .. .d .--. . . . . .... -----. .._.-._ . .. :---.. .: .... ._... . . . . ... . . . .-........ - ...- .....- . . . .I .--- __. _h.. h u y c i E a t .Ma .. _-- ....-..--.. . . .- ..... -. .. ,. . Page27~f38 . : .. .- &ire.Ahalytical Laboratories,I~C. ' Page 88 of 102 . . . _. Centre Stu. dy. No..:023-063 . . . . . i . ,I ' .. I . ... I r ?. I . . .. e . - . I .' i . .. .TIB-I----S - .. - .. . .. - .. .. - .. e - . .. ..... .. .. .. .- . - .... -. .. ;'' j '.j . . c' - .- . ' -. .- . . - _ % . . . .,.-. . .... I.. .- . .I . . . 0 .'. . . . . . . . . . .. e. . .* . I ' .- -. . . -'. .* .. .. .. . - .. - .. .. -.._ .. -Cent&.Analytical.Labomtdries~, nc...... .. . . . - ... - Page 89.of io2 Centre Study No.:023-063 .. I .... . . ......... . ..- _. . . . . . . . _._. ._... . l." e.---. .. ._ ........ . . t ._-.-..--.-.. .. .' - .--.. .-I-. . . ------ cz.nfr.-ek w o l '-L No.o o .. ::-- p-M3 4 j " 3 . ... a. -_ . ' .-. -------.- - . ----.. . ~ .' ,. .. I. . % ... ... .. .. .. .. .. . . . . ..... . . .. . . . . . . . . Page29of38 . .. . .' Page 90 of 102 Centre Study No.:023-063 ...... -- , *. ,_ ..- .. Centre Study No.: 023-063 . .. -.-. .. .. .- . . .. . . . . . .*.# .$. . .- I' .. e . .. .- . ...._- ..' Centrel hdytical Laboratories,Inc. .. .- .. .I .............. Page 92 of 102 Centre StudyNo.:023-063 . -. . --_ -.,'i I ...... .:.--I_ ... ......--.I.,................-.-.. .... --- ....... ....-._..._. _____...._- i 2- I .. .. .. -. .... . -. .. . I * . -r c . . !. - . I. -. I ... .. .. ... . - - .. * . . . .. - ... . - ... .- . . . . . . -I .. .. . . . . .. . .. .. . .. - - .. .. .. .* .. *.. . .. .. .. -... . . . ..-. .. .. ........ . . . . . . . ... . .. .. .-.._. Centre Study No.:023-063 . . . ..................... --' ................. . ...... --- ... -....... 1 .. .;-....--. .... ......... &-&*-&@o,No.w&.....036.363.. .* .. . Y .- . - .6 . . .. . . .- . . ... .. .. .. --e .. .. .. .. * . .. . .. ... - . . . . .- . . .. . . . .. . ! .. ..: .. .. .- I .\-; ! . . *Awdjlidr.L.i*#m1J -- ... . . ... . ... __.__........... 7 I -.--_-..... Cen~.:.AnalflcaLl aboratories,Inc. , .. e . . . - . '. *. .. - .-..-- . ... +a. . .. . .. .. ... . -330f38 ............ .. . . . - . Page94 of 102 .......... ....... ..... ....... . _. . . Centre Study No.:023-063 . ...... *-.....-..... ------. ,. I ..-., . ....... I. -- . .__..._....-........................ ....&fi-RO!&OI NO.00pM3463 .- -. . ."_.". . ..-----. - . . . . .- __ _-.-.-.-_-...-.......... .---..--_ .-.__ I * .-. - . -e-- -----._. I .. .. . . ..- -.. * .. .. . . c . .. .. .' .. I .... . . . - - .. I .. b-. ' ..... ......._. . . .. . . . .^---n--. . . . .*----.--.- 24 . ... .-.--.. . . . ..-- ..... .......- . . . . . . . . . . . . Centre Study No.:023-063 --.-...-. ......... i t ' .. e-. j I :. . ...._. ....-....... ......-..--. *. ... . *--. .- .. . .-- '?':''&i~-~brNoo:opM3-063 ... * . .- : i- ...-. ............. . . . - . - - -.-----.-: . ^._.__1.. i ' ........ , .. .. .- .. ... . ... e . .. . .. .. ..::I . . ... .. .. I. .. .. e . ................. .. ...... ...---.__ ._ --- . .- . "":'&fitreAnalyticalLaboratories, bc. . . .. . . ... ..-. . . . . . Page%of 102 e Centre Study No.:023-063 I ._.._.. . I I . '.. :. .. .- -. - _.. .i . ... ...... -...-....-. .-.--:117.. . . . . . -. .. I . . .--c.areMctaoa No:ooM.fJ~.o~; I I -. ' .. I. . . . -. .. ._ .. .. _. -- . . . . .. . . . .' . .... z. . .. I I . . ..-. .... . ... . . . :. .. .. -. .. . Page36of38 . Page 97 of 102 Centre Study No.:023-063 . . . . -. - .. . - . .. . .. .. .. .. ... ... - . . .. .... . . - - .. _ . .: . .. . -. :I 2 - .. .- 2 i .. .. --.i - ~~~jnstudy?oliou . . . . . ..._.* . . . . . . ...... .. ....... . . .. ' antre Analytica~iabomtories,~nc; . .. .". . . . ... . .. pq: 3 ............. .. .. . page 370f 38 . Page 98 of 102 Centre Study No.: 023-063 -. ... .-Y '> .. ... . ... "..... .. .. .. ... I I 1 ... .. .. . . .. - .. .- .: .. -- -. . . . .+= e..! - '5 ... *i r. - .. : . .. .. . ... .- .---.::._ .-...-__-- .-.-..-..- .............. ..i" . .U... . .- . ---.---. ..._ .. Page 38 of 38 _.. Centre hdyticd Laboratories, Inct . Page 99 of 102 Centre StudyNo.:023-063 mfii!!\ CEntre Analytical LaboratoriEs, Inc, 3048 Racpreh Drive, State College, PA 16801. Phone: (814)231-8032,Faaimile: (814)231-1253 Pase 1 of 1 PROTOCOL AMENDMENT Amendment Number: 1 11Centre StudyNumber:023-063 EffectiveDate: 3/23/01 Centre Protocol Number: OOP-023-063 DESCRIPTIONOF AMENDED SECTION i 1. Title 2. 0 8 Analytical Method 4.4.4.f. II AMENDEDTO 1. Change the analytical phase title to read Extractionof PotassiumPerfluorooctanesulfonatefromEgg Membrane,Albumen, and Yolk forAnalysisUsing HPLC-ElectrospraYl'Mass Spectrometry(This will also change I on Page 2 and Page 3) 2. Change the first sentence to read Fortificationrecoverieswithin 70-12046 are-acceptablefor fortificationsat the LOQ 1. Per Sponsorrequest to maintain consistencywith method. 2. Reflectthe requirementsthat were followed duringthe method validation. IMPACTONTHESTUDY 1-2. No negative impact on the study. Study Director Signature Date / d CentreQAUReview h 6/d7b I Febwry 12.1998/1 Centi.e.Arralytica1Laboratories, Inc; .. Page 100 of 102 Centre Study No.: 023.-063 mti!!\ CcntrE Analytical Laboratories, Inc. PROTOCOLAMENDMENT Amendment Number 2 Effective Date: 3/26/01 Page 1of; Centre StudyNumber.023-063 Centre Protocol Number: OOP-023-063 DESCRIPTION OF AMENDED SECTION 1. 9 Section 9 Analytical Method 2. 9 Section 9: AnalyticalMethod 3. 9 Section 1 0 ExperimentalDesign I' AMENDEDTO 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 ASTMType I H20 and patted dry. 2. On Page 24 Section 4.4.4.d, remove: One ASTM Type1water blank 3. Amend Page dlast paragraph,second sentenceto read - - For egg yolk and albumen, 0.5 g samplesand eggmembrane 0.1 g sampleswill be - weighed and dried in an oven at 105Cfor-- 12-f4 hours. It 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 sampleis only to check forPFOS residue present in the ASTMType I H20used. 3. To account for sample availability. IMPACTONTHESTUDY 1It1 1-3. No negativeimpact on the study. JL/e(, 1. b U Study Director Signature GlSla\ Date / / Centre Analytical Laboratories, Inc. Centre QAU Review Qk 4l;L7/p/ February 12, 199811 Page 101of 102 Centre Study No.: 023-063 PROTOCOL DEVIATION Deviation Number: 1 Date of Occurrence: (1) 03/17/01. (2) 04102/01 1. 8 8 AnalyticalMethod 4.4.43 Quantified Centre sample0103196 and its duplicate injectionwhen the concentrations wen greater than that of the highest standardin !kt 032601C 2. 8 Amendment #1: Accepted recoveries of 58% and 61% for Centre sample 0003800 Spk A and its duplicate injection in Set 033001C. 1-2. Protocol deviation issued. 1. No negative impact because the areas were c 5% greater than the highest standard. 2. No negativeimpact because Spk B gave acceptablerecoveriesand sample resultsreported in the set were only for confirmation. I spa- Date & 'Centre Analytical Laboratories, Inc. Page 102 of 102