Document bBGjz4NEXZqNRpJZ896pO6a7Z

STU D Y T IT L E Extraction of Potassium Perfluorooctanesulfonate from Egg Membrane, Albumen, and Yolk for Analysis Using HPLC-Electrospray/Mass Spectrometry DATA REQUIREMENTS Analytical Method Requirements STU D Y D IR EC TO R Sean Gallagher STUDY COM PLETED ON September 06,2001 PER FO R M IN G LA BO R A TO R Y / TE ST IN G FA C IL IT Y 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 PRO JECT Centre Protocol Number: 00P-023-063 Centre Study Number: 023-063 Total Pages: 102 000243 Cantre Study No.: 023-063 I Centre Study Num ber 0 2 3-069. entitled "Extraction o f Potassium Perfluorooetanw ulfonatft from E gg M em brane, A lbum en, and Y olk fo r A nalysis U sing H PLC -Eleccfoipray/M asa Spectrom etry,'* conducted for 3M Environm ental T echnology and Safety Service, w as perform ed in com pliance w ith U S EPA T SC A G ood Laboratory P ractice Standards (4 0 CFR Part 792) by Centre A nalytical Laboratories, Inc w ith the follow in g exceptions. 1. 1792.63. M aintenance and Calibration o f E quipm ent N o equipm ent SO P wag in p lace for the bead beater u sed in the extraction at the tim e o f use. C entr A nalytics] Laboratories, Inc. Study U iitcto r W ild life International, Ltd. Centre Analytical Laboratori, In. Page 2 of 102 000244 C entre Study N o.: 023-063 QUALITY ASSURANCE STATEMENT Centre Analytical Laboratories' Quality Assurance Unit reviewed Centre Study Number 023-063, entitled, "Extraction of Potassium Perfluorooctanesulfonate from Egg Membrane, Albumen, and Yolk for Analysis Using HPLC-Electrospray/Mass Spectrometry". All examined phases were reviewed for conduct according to Centre Analytical Laboratories' Standard Operating Procedures, the Study Protocol, and all applicable Good Laboratory Practice Standards. All findings were reported to the Study Director and to management. Date Reported to Date Repeated to Date Centre Study Director and iDSBgctsd Management SptmafliManagsitnent 1. Protocol Review 3/30/01 4/9/01 5/7/01 2. Extraction, Fortification 3/30/01 4/9/01 5/7/01 3. Raw Data Review 4/5-9/01 4/18/01 5/7/01 4. Raw Data and Draft Report Review 4/17-18/01 4/26/01 5/7/01 5. Final Report Review 6/28/01 9/6/01 9/6/01 Date C entre A nalytical Laboratories, Inc. Page 3 o f 102 000245 C ent Study No.; 023-063 CBS mmQfAvwmism T h il report, for Centre Study Number 0234)63, it a true and com plete representation of the a w data for the study- Subm itted by; Centre Analytical Laboratories, Inc. 3048 Research Drive State C ollege, PA 16801 (814)231*8032 Principal Investigator, Centre: S c ie n tist Centre Analytical Laboratories, Inc, Centre A nalytical L&boratoriee, Inc. F acility Management: Richard A in ^ h j* P r e sid e n t Centre A nalytical Laboratories, Inc. Study Director d if* q -a u x -o i D ate ^ --------- Seen Gallagher W ild life international, Ltd. * Sponsor R epresentative I Centre Analytical Laboratories, Inc. Pag* 4 o f 102 000246 Centre Study No.: 023-063 STUDY IDENTIFICATION "Extraction of Potassium Perfluorooctanesulfonate from Egg Membrane, Albumen, and Yolk for Analysis Using HPLC-Electrospray/Mass Spectrometry," CENTRE PROTOCOL NUMBER: 00P-023-063 CENTRE STUDY NUMBER: 023-063 TYPE OF STUDY: Analytical SAMPLE MATRIX: Mallard and Quail Egg Membrane, Albumen, and Yolk TEST SUBSTANCE: Perfluorooctanesulfonate (PFOS) SPONSOR: 3M Environmental Technology and Safety Services Building 2-3E-09 PO Box 33331 St. Paul, MN 55133-3331 STUDY DIRECTOR: Sean Gallagher W ildlife International, Ltd. 8598 Commerce Drive Easton, MD 21601 TESTING FACILITY: Centre Analytical Laboratories, Inc. 3048 Research Drive State College, PA 16801 ANALYTICAL PHASE TIMETABLE: Study Initiation Date: Experimental Start Date: Experimental Termination Date: Study Completion Date: 03/26/01 03/26/01 04/02/01 09/06/01 C entre A nalytical L aboratories, Inc. Page 5 of 102 000247 C entre 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 Arjmand 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 A nalytical Laboratories, Inc. Page 6 of 102 000248 C entre Study No.: 023-063 TABLJS.QF QM JBTOnS Page TITLE PA G E.......................................................................................................................... 1 GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT.............................. 2 QUALITY ASSURANCE STATEMENT............................................................................ 3 CERTIFICATION OF AUTHENTICITY............................................................................. 4 STUDY IDENTIFICATION.................................................................................................. 5 PROJECT PERSONNEL....................................................................................................... 6 TABLE OF CONTENTS....................................................................................................... 7 LIST OF TABLES..................................................................................................................8 LIST OF FIGURES...............................................................................................................11 LIST OF APPENDICES...................................................................................................... 12 1.0 SUMMARY...................................................................................................................13 2.0 OBJECTIVE...................................................................................................................13 3.0 INTRODUCTION..........................................................................................................13 4.0 TEST SYSTEM .............................................................................................................14 5.0 REFERENCE MATERIAL.......................................................................................... 14 6.0 DESCRIPTION OF ANALYTICAL METHOD..........................................................15 6.1 Extraction Procedure................................................................................................... 15 6.2 Preparation of Standards and Fortification Solutions................................................ 15 6.3 Chromatography...........................................................................................................16 6.4 Instrument Sensitivity................................................................................................. 16 6.5 Description of Instrument and Operating Conditions................................................ 17 6.6 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 C entre A nalytical L aboratories, Inc. Page 7 o f 102 000249 C entre Study N o.: 023-063 Table I. LIST OF TABLES Page Summary of PFOS in Reagent Blanks..............................................................23 Table EL Summary o f PFOS in Mallard Membrane B lanks...........................................24 Table m . Summary o f PFOS in M allard Albumen Blanks.............................................. 24 Table IV. Summary of PFOS in Mallard Yolk B lanks.................................................... 24 Table V. Summary of PFOS Recoveries in M allard Membrane.....................................25 Table VI. Summary of PFOS Recoveries in M allard Albumen.......................................25 Table VEL Summary of PFOS Recoveries in M allard Yolk..............................................26 Table VIIL Summary of PFOS for Week 1 M allard Membrane Control Samples............26 Table IX. Summary o f PFOS for Week 1 M allard Membrane Samples at 6.2 ppm ai....................................................................................................... 27 Table X. Summary of PFOS for Week 1 M allard Membrane Samples at 17.6 ppm ai..................................................................................................... 27 Table XI. Summary of PFOS for Week 1 M allard Albumen Control Sam ples..............28 Table XU. Summary of PFOS for Week 1 Mallard Albumen Samples at 6.2 ppm a i.... 28 Table XIH. Summary of PFOS for Week 1 M allard Albumen Samples at 17.6 ppm a i.. 29 Table XIV. Summary of PFOS for Week 1 M allard Yolk Control Sam ples.....................29 Table XV. Summary of PFOS for Week 1 Mallard Yolk Samples at 6.2 ppm a i........... 30 Table XVI. Summary o f PFOS for W eek 1 M allard Yolk Samples at 17.6 ppm a i ......... 30 Table XVEL Summary of PFOS for Week 6 M allard Membrane Control Samples..........31 Table XVIII. Summary of PFOS for Week 6 M allard Membrane Samples at 6.2 ppm a i...................................................................................................... 31 Table XIX. Summary of PFOS for Week 6 M allard Membrane Samples at 17.6 ppm ai...................................................................................................... 32 Table XX. Summary of PFOS for Week 6 M allard Albumen Control Sam ples..............32 Table XXI. Summary of PFOS for Week 6 M allard Albumen Samples at 6.2 ppm a i.... 33 Table XXII. Summary of PFOS for Week 6 M allard Albumen Samples at 17.6 ppm a i. 33 Table XXUL Summary of PFOS for Week 6 Mallard Yolk Control Sam ples........... 34 Table XXIV. Summary of PFOS for Week 6 Mallard Yolk Samples at 6.2 ppm a i.. 34 C entre A nalytical L aboratories, Inc. Page 8 o f 102 ooozso C entre Study N o.: 023-063 Table XXV. Summary of PFOS for Week 6 Mallard Yolk Samples at 17.6 ppm a i.... 35 Table XXVI. Summary of PFOS in Quail Membrane B lanks.................................... 35 Table XXVII. Summary of PFOS in Quail Albumen B lanks............................................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 XXXI. Summary of PFOS Recoveries in Quail Yolk..............................................37 Table XXXH. Summary of PFOS for Week 1 Quail Membrane Control Sam ples......... 38 Table XXXIII. Summary of PFOS for Week 1 Quail Membrane Samples at 6.2 ppm a i....................................................... 38 Table XXXIV. Summary of PFOS for Week 1 Quail Membrane Samples at 17.6 ppm a i................................................................................................... 39 Table XXXV. Summary of PFOS for Week 1 Quail Albumen Control Sam ples............39 Table XXXVI. Summary of PFOS for Week 1 Quail Albumen Samples at 6.2 ppm ai... 40 Table XXXVII. Summary of PFOS for Week 1 Quail Albumen Samples at 17.6 ppm a i.................................................................................................40 Table XXXVIII. Summary of PFOS for Week 1 Quail Yolk Control Samples................41 Table XXXEX. Summary of PFOS for Week 1 Quail Yolk Samples at 6.2 ppm a i........ 41 Table XL. Summary of PFOS for Week 1 Quail Yolk Samples at 17.6 ppm a i............. 42 Table XLL Summary of PFOS for Week 6 Quail Membrane Control Samples............... 42 Table XLIt. Summary of PFOS for Week 6 Quail Membrane Samples at 6.2 ppm a i..... 43 Table XLm. Summary of PFOS for Week 6 Quail Membrane Samples at 17.6 ppm a i.. 43 Table XUV. Summary of PFOS for Week 6 Quail Albumen Control Samples.............. 44 Table XLV. Summary of PFOS for Week 6 Quail Albumen Samples at 6.2 ppm a i...... 44 Table XLVI. Summary of PFOS for Week 6 Quail Albumen Samples at 17.6 ppm a i.... 45 Table XLVII. Summary of PFOS for Week 6 Quail Yolk Control Sam ples................... 45 Table XLVin. Summary of PFOS for Week 6 Quail Yolk Samples at 6.2 ppm a i......... 46 Table EL. Summary of PFOS for Week 6 Quail Yolk Samples at 17.6 ppm ai.............. 46 Table L. Summary of Moisture Determinations for M allard Membrane Samples....... 47 Table LL Summary of Moisture Determinations for M allard Albumen Samples......... 48 C entre A nalytical L aboratories, Inc. Page 9 o f 102 000251 C entre Study N o.: 023-063 Table LH. Summary of Moisture Determinations for Mallard Yolk Samples............... 49 Table LD0L Summary of Moisture Determinations for Quail Membrane Sam ples......... 50 Table LTV. Summary of Moisture Determinations for Quail Albumen Samples............ 51 Table LV. Summary of Moisture Determinations for Quail Yolk Samples....................52 C entre A nalytical Laboratories, Inc. Page 10 o f 102 000252 C entre Study N o.: 023-063 Figure 1. LIST OF FIGURES Page Typical Calibration Curve for PFOS............................................................... 54 Figure 2. Chromatogram Representing a 0.1 ng/mL standard for PFO S......................55 Figure 3. Chromatogram Representing a 1.0 ng/mL standard for PFO S......................56 Figure 4. Chromatogram Representing a Reagent Blank for PFOS (Centre ID Reagent Blank A, Set: 032601A).................................................. 57 Figure 5. Chromatogram Representing Control Mallard Membrane for PFOS (Centre ID: 0003802 Blank A, Set: 032601C)............................................................. 58 Figure 6. Chromatogram Representing Control Mallard Membrane Fortified with 10 ppb of PFOS, (Centre ID: 0003802 Spk A Dup. Inj., Set: 032601C)............59 Figure 7. Chromatogram Representing Mallard Membrane Sample from W eek 1 at 6.2 ppm ai (E01-0263-20825) (Centre ID: 0103092, Set: 032601C)........ 60 000253 C entre Study N o.: 023-063 LIST OF APPENDICES Page Appendix A Study Protocol OOP-023-063 (Centre Study No. 023-063) and Amendments and Deviation........................................................................61 C entre A nalytical Laboratories, Inc. Page 12 o f 102 000254 C entre 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 00P-023-063 (Appendix A). The limit of quantitation for each matrix was 10 ppb. The LOQ for each matrix was determined in a method validation study performed at Centre (Centre Study: 023-027). PFOS ranged from non-quantifiable (where non-quantifiable is defined as less than the concentration of the lowest calibration standard (0.1 ng/mL)) levels to 434 ng/g in the mallard membrane samples, from non-quantifiable levels to 3550 ng/g in the mallard albumen samples, and from non-quantifiable levels to 63,500 ng/g in the mallard yolk samples. PFOS ranged from non-quantifiable levels to 20,900 ng/g in the quail membrane samples, from non-quantifiable levels to 93.9 ng/g in the quail albumen samples, and from non-quantifiable levels to 65,500 ng/g in the quail yolk samples. All of these results are reported on a wet weight basis. The average percent recoveries standard deviations for PFOS in mallard membrane, albumen, and yolk samples were 91% 6%, 91% 16%, and 93% 13%, respectively. The average percent recoveries standard deviations for PFOS in quail membrane, albumen, and yolk samples were 102% 11% , 85% 5%, and 96% 7%, respectively. 2.0 OBJECTIVE The objective of this analytical phase of the study was to determine levels of perfluorooctanesulfonate (PFOS) in specimens of mallard and quail egg membrane, albumen, and yolk using the analytical method titled, "Determination of Perfluorooctanesulfonate in Egg Membrane, Ablumen and Yolk by LC/MS/MS". 3.0 INTRODUCTION This report details the results of the analysis for the determination of PFOS in mallard and quail egg membrane, albumen, and yolk using the analytical method entitled, "Determination of Perfluorooctanesulfonate in Egg Membrane, Ablumen and Yolk by LC/MS/MS". This analytical portion of the study was initiated on March 26, 2001, when the study director signed Centre protocol number 00P-023-063. The experimental start date was March 26,2001, and the experimental termination date was April 2,2001. C entre A nalytical Laboratories, Inc. Page 13 o f 102 000255 C entre 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 W ildlife International Ltd., Easton, MD and stored frozen at Centre. The following Centre ID's were assigned to the different matrices: M ate Cep tte P M allard Membrane 0003802 M allard Albumen 0003800 M allard Yolk 0003799 Quail Membrane 0003798 Quail Albumen 0003796 Quail Yolk 0003795 On February 23, 2001, 308 quail egg samples were received. The samples were stored frozen until they were logged in by Centre personnel on February 26, 2001 and stored frozen. On February 24, 2001, 194 mallard egg samples were received. The samples were stored frozen until they were logged in by Centre personnel on March 1, 2001 and stored frozen. Each egg sample received was divided into membrane, albumen, yolk, and shell. Each component of the egg was given a unique Centre ID at the time of log-in. Although all of the egg samples were received at Centre, only those designated by the sponsor were extracted and analyzed. Sample log-in and chain of custody information can be found in the raw data package associated with this study. Storage records will be kept at Centre Analytical Laboratories, Inc. and a true copy of the storage records can be found in the raw data package associated with this study. 5.0 R EFER EN C E M A TER IA L The analytical standard PFOS was received at Centre on June 3, 2000 from 3M Environmental Technology and Services. Stability was not assessed in this study but the reference material was stored according to the certificate of analysis (frozen) and was used prior to expiration. The solubility of the reference material was not relevant to this study. The available information for the reference material is listed below. Compound PFOS Centre Control No.. TCR No. Purity (%) Expiration Date 00-023-042 TCR 00017-46 97.9 08/31/01 Centre A nalytical L aboratories, Inc. Page 14 o f 102 000256 C entre Study N o.: 023-063 The molecular structure of PFOS is given below. PFOS Chemical Name Molecular weight O 'IC8F17S On- = = Perfluorooctanesulfonate 499 (CgFnSOs-) Note: The neutral molecule and standard from which the PFOS (anion) is obtained, is potassium perfluorooctanesulfonate [C8F17SO3K], molecular weight 538. 6.0 DESCRIPTION OF ANALYTICAL METHOD Analytical method entitled "Determination of Perfluorooctanesulfonate in Egg Membrane, Ablumen and Yolk by LC/MS/MS" was used for this study. 6.1 E xtraction Procedure Egg mem brane A 0.1 g sample was used for the extraction procedure. One m illiliter 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 m illiliters 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 m illiliter 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 P reparation of S tandards and Fortification Solutions Standard solutions were prepared on March 14, 2001 as specified in Centre Analytical Laboratories' protocol 00P-023-027. An individual stock standard solution of PFOS was prepared at a concentration of 100 pg/mL by dissolving 10 mg of the standard (corrected for purity and salt content) in methanol. From this solution, a 1.0 pg/mL fortification standard solution was prepared by taking 1 mL of the stock and bringing the volume up to 100 mL with methanol. Centre Analytical Laboratories, Inc. Page 15 of 102 000257 C entre Study No.: 023-063 The 0.1 (ig/niL fortification standard was prepared by taking 10 mL of the 1.0 pg/mL fortification standard and bringing the volume up to 100 mL with methanol. A 0.01 pg/mL standard was prepared by taking 10 mL of the 0.1 pg/mL standard and bringing to 100 mL with methanol. A set of standards containing PFOS was prepared by dilution of the 0.1 pg/mL and 0.01 pg/mL solutions in the following manner. Initial Cone. (pg/mL)1 0.1 0.1 0.1 0.01 0.01 0.01 1of PFOS Volume (mL) 5 2 1 5 2 1 Diluted to (mL) 100 100 100 100 100 100 Final Cone. (pg/mL) 0.005 0.002 0.001 0.0005 0.0002 0.0001 The stock standard solution and all fortification and calibration standard solutions were stored in a refrigerator (4C 2C) when not in use. Documentation of standard preparation can be found in the raw data associated with this report. 6 3 Chromatography Quantification of PFOS was accomplished by LC/MS/MS electrospray. The retention time of PFOS was ~ 4.4 min. Peaks were detected in the control matrices corresponding to the analyte retention time, but the amounts detected were only significant enough to alter the results in several o f the recoveries reported and the rest were less than the low est calibration standard (0.0001 pg/mL). 6.4 Instrument Sensitivity The smallest standard amount injected during the chromatographic run had a concentration of 0.0001 pg/mL of PFOS. C entre A nalytical Laboratories, Inc. Page 16 o f 102 000258 C entre Study No.: 023-063 6.5 Description of Instrument and Operating Conditions Instrument: PE SCIEX API 3000 Biomolecular Mass Analyzer Interface: SCIEX Turboion Spray Liquid Introduction Interface Computer Dell UltraScan P I 110 Software: HPLC: PE SciexAnalyst version 1.1 Windows NT, version 4 Hewlett Packard (HP) Series 1100 HP Quat Pump HP Vacuum Degasser HP Autosampler HP Column Oven HPLC Column:Genesis Cg (Jones Chromatography), 2.1 mm x 50 mm, 4p Column Temp.: 35 C Injection Vol.: 10 pL Mobile Phase (A): 2 mM Ammonium Acetate in ASTM type I water Mobile Phase (B): Methanol Flow Rate: 0.3 mL/min. Time % A % B 0 60 40 1.0 0 100 7.0 0 100 7.5 60 40 11.0 60 40 Ions monitored: Analvte PFOS Mode Transition Monitored negative 499 - 99 Approximate Retention Time (nain) 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 C entre A nalytical Laboratories, Inc. SI -4200.0 -51.0 -230.0 10.0 -70 -5 Page 17 o f 102 000259 C entre Study N o.: 023-063 DF-Deflector CEM-Channel Electron Multiplier 300.0 2800.0 Gas Flows Nebulizer Gas Curtain Gas Collision Gas TIS Temperature Set 12 14 4 350C 6.6 Quantitation and Example Calculation Ten microliters of sample or calibration standard were injected into the LC/MS/MS. The peak area was measured and the standard curve was generated (using 1/x weighted linear regression) by Analyst software using at least five concentrations of standards. The concentrations for mallard and quail egg membrane, albumen, and yolk (ng/g) were determined from the equations below. Equation 1 calculated the amount of analyte found (in ng/mL, based on peak area) using the standard curve (linear regression parameters) generated by the Analyst software program. Then Equation 2 calculated the amount of analyte found in ng/g. Equation 1: Analyte found (ng/mL) = (Peak area - intercept) slope Equation 2: Analyte found (ng/g) = anal, found (nf/m Ll x FV (mL) x DF sample wt. (g) Where: FV = Final Volume DF = Dilution Factor For samples fortified with known amounts of PFOS prior to extraction, Equation 3 calculated the percent recovery. Equation 3: Recovery (%) = (anal. Found (ng/g) - avg. anal, in Ctrl (ng/g) x 100 amount added (ng/g) An example of a calculation using an actual sample follows: M allard albumen sample Centre ID 0003800 Spk A (Set: 032601A), fortified at 10 pbb with PFOS, where: C entre A nalytical Laboratories, Inc. Page 18 o f 102 000260 C entre Study N o.: 023-063 peak area intercept slope dilution factor ng/g added (fort level) avg. amt in controls final volume sample weight = = = = = = = = 3369 971 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) = [336r 971] 6700 2 From equation 2: Analyte found (ng/g) = 0.36 ng/mL 0.36 ng/mL x 25 mL x 1 ) 1.0038 (g) From equation 3: % Recovery 2 8.97 ng/mL = (8.97 n g /e x - 0 ne/g)x 100 10 ng/g 2 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 EXPERIMENTAL DESIGN 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 IH 2O blank was analyzed. C entre A nalytical L aboratories, Inc. Page 19 o f 102 000261 C entre Study No.: 023-063 8.0 RESULTS PFOS found in the reagent blanks are listed in T able I. The PFOS in the mallard membrane, albumen, and yolk blanks are given in Tables II-IV . The PFOS in the quail membrane, albumen, and yolk blanks are given in Tables XXVT-XXVIIL PFOS was detected in several of the control matrices, however few were significant enough to alter the recoveries reported. Individual recoveries for PFOS in the mallard membrane, albumen, and yolk samples are detailed in Tables V-VII. The average percent recoveries standard deviations for PFOS in mallard membrane, albumen, and yolk samples were 91% 6%, 91% 16%, and 93% 13%, respectively. Individual recoveries for PFOS in the quail membrane, albumen, and yolk samples are detailed in T able XXIX-XXXI. The average percent recoveries standard deviations for PFOS in quail membrane, albumen, and yolk samples were 102% 11% , 85% 5%, and 96% 7%, respectively. PFOS ranged from non-quantifiable (where non-quantifiable is defined as less than the concentration of the lowest calibration standard (0.1 ng/mL)) levels to 434 ng/g in the mallard membrane samples, from non-quantifiable levels to 3550 ng/g in the mallard albumen samples, and from non-quantifiable levels to 63,500 ng/g in the mallard yolk samples. These results are reported on a wet weight basis. Individual results are listed in Tables VIII-XXV. Some of the high levels of PFOS reported for some of the mallard egg membrane and egg albumen samples could be from possible contamination from the egg yolk or outer shell. PFOS ranged from non-quantifiable levels to 20,900 ng/g in the quail membrane samples, from non-quantifiable levels to 93.9 ng/g in the quail albumen samples, and from nonquantifiable levels to 65,500 ng/g in the quail yolk samples. These results are reported on a wet w eight basis. Individual results are listed in T ables X X X II-IL. Some o f the high levels of PFOS reported for some of the quail egg membrane and egg albumen samples could be from possible contamination from the egg yolk or outer shell. The Sponsor Representative also informed Centre that the quail outer shells were not washed prior to separation of the internal components. The average percent moistures for the mallard and quail egg membrane, albumen, and yolk samples are given in Tables L-LV. 9.0 CONCLUSIONS The mallard and quail egg membrane, albumen, and yolk samples were successfully extracted and analyzed according to protocol 00P-023-063. C entre A nalytical Laboratories, Inc. Page 20 o f 102 000262 C entre Study No.: 023-063 10.0 RETENTION OFDATA 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 o f temperature logs will be submitted. Exact copies of all raw data, as well as a signed copy o f 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. C entre A nalytical Laboratories, Inc. Page 21 of 102 000263 C entre Study No.: 023-063 TABLES Centre A nalytical Laboratories, Inc. Page 22 of 102 000264 C entre Study N o.: 023-063 Table I. Summary of PFOS in Reagent Blanks Sponsor ID na na na na na na na na na na na na na na na na na na na na na na na na na na Centre ID Reagent Blank A Reagent Blank A* Reagent Blank A Reagent Blank A* Reagent Blank A Reagent Blank A* Reagent Blank A Reagent Blank A* Reagent Blank A Reagent Blank A* Reagent Blank A Reagent Blank A* ReagentBlank A Reagent Blank A* Reagent Blank A Reagent Blank A* Reagent Blank A Reagent Blank A* Reagent Blank A Reagent Blank A* Reagent Blank A Reagent Blank A1" Reagent Blank A Reagent Blank A* Reagent Blank A Reagent Blank A* * Duplicate injection Set Number 032601A 032601A 032601B 032601B 032601C 032601C 032701A 032701A 032701B 032701B 032701C 032701C 032801A 032801A 032801B 032801B 032901A 032901A 032901B 032901B 033001A 033001A 03300IB 033001B 033001C 033001C Extraction Analysis Date Date 3/26/01 3/26-27/01 3/26/01 3/26-27/01 3/26/01 3/27/01 3/26/01 3/27/01 3/26/01 3/27/01 3/26/01 3/27/01 3/27/01 3/28/01 3/27/01 3/28/01 3/27/01 3/28-29/01 3/27/01 3/28-29/01 3/27/01 3/29/01 3/27/01 3/29/01 3/28/01 3/30/01 3/28/01 3/30/01 3/28/01 3/30/01 3/28/01 3/30/01 3/29/01 3/31/01 3/29/01 3/31/01 3/29/01 3/31-4/1/01 3/29/01 3/31-4/1/01 3/30/01 4/1/01 3/30/01 4/1/01 3/30/01 4/2/01 3/30/01 4/2/01 3/30/01 4/2/01 3/30/01 4/2/01 AVERAGE: STANDARD DEVIATION: Analyte Found(nnb) NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) LOQ = 10 ppb C entre A nalytical Laboratories, Inc. Page 23 o f 102 000265 C entre Study No.: 023-063 Table II. Summary of PFOS in Mallard Membrane Blanks Sponsor ID na na na na Centre m 0003802 Blank A 0003802 Blank A* 0003802 Blank A 0003802 Blank A* Set Number 032601C 032601C 032701C 032701C Extraction Analysis Date Date 3/26/01 3/27/01 3/26/01 3/27/01 3/27/01 3/29/01 3/27/01 3/29/01 AVERAGE: STANDARD DEVIATION: Analyte Found (ppb) NQ NQ NQ NQ NQ NQ Table m . Summary of PFOS in Mallard Albumen Blanks Sponsor ID na na na na na na Centre ID 0003800 Blank A 0003800 Blank A* 0003800 Blank A 0003800 Blank A* 0003800 Blank A 0003800 Blank A* Set Number 032601A 032601A 032701B 032701B 033001C 033001C Extraction Analysis Date Date 3/26/01 3/26/01 3/27/01 3/26-27/01 3/26-27/01 3/28-29/01 3/27/01 3/28-29/01 3/30/01 3/30/01 4/2/01 4/2/01 AVERAGE: STANDARD DEVIATION: Analyte Found (ppb) NQ NQ NQ NQ 6.03 6.59 2.94 2.62 Table IV. Summary of PFOS in Mallard Yolk Blanks Sponsor ID na na na na Centre ID 0003799 Blank A 0003799 Blank A* 0003799 Blank A 0003799 Blank A* Set Number 032601B 032601B 032701A 032701A Extraction Analysis Date Date 3/26/01 3/27/01 3/26/01 3/27/01 3/27/01 3/28/01 3/27/01 3/28/01 AVERAGE: STANDARD DEVIATION: Analyte Found(ppb) NQ NQ 9.57 9.80 5.48 4.87 * Duplicate injection NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) LOQ = 10 ppb For residues recorded as NQ, half of the concentration of the lowest standard (1.25 ppb) was used to calculate the average and standard deviation. C entre A nalytical L aboratories, Inc. Page 24 o f 102 C entre Study No.: 023-063 Table V. Summary of PFOS Recoveries in Mallard Membrane Sponsor ID na na na na na na na na Centre ID 0003802 Spk A 0003802 Spk A* 0003802 Spk B 0003802 Spk B* 0003802 Spk A 0003802 Spk A* 0003802 Spk B 0003802 Spk B* Set Number 032601C 032601C 032601C 032601C 032701C 032701C 032701C 032701C Extraction Analysis Amt Date Date Added(ppb) 3/26/01 3/26/01 3/27/01 3/27/01 10 10 3/26/01 3/26/01 3/27/01 3/27/01 100 100 3/27/01 3/27/01 3/27/01 3/27/01 3/29/01 3/29/01 3/29/01 3/29/01 10 10 97 97 AVERAGE: STANDARD DEVIATION: RELATIVE STANDARD DEVIATION: % Recovery 86 87 87 81 97 94 95 98 91 6 7 Table VI. Summary of PFOS Recoveries in Mallard Albumen Sponsor ID na na na na na na na na na na na na Centre ID 0003800 Spk A 0003800 Spk A* 0003800 Spk B 0003800 Spk B* 0003800 Spk A 0003800 Spk A* 0003800 Spk B 0003800 Spk B* 0003800 Spk A 0003800 Spk A* 0003800 Spk B 0003800 Spk B* Set Number 032601A 032601A 032601A 032601A 032701B 032701B 032701B 03270IB 0330Q1C 033001C 033001C 033001C Extraction Analysis Amt Date Date Added (ppb) 3/26/01 3/26-27/01 10 3/26/01 3/26-27/01 10 3/26/01 3/26-27/01 99 3/26/01 3/26-27/01 99 3/27/01 3/28-29/01 10 3/27/01 3/28-29/01 10 3/27/01 3/28-29/01 99 3/27/01 3/28-29/01 99 3/30/01 4/2/01 10 3/30/01 4/2/01 10 3/30/01 4/2/01 98474 3/30/01 4/2/01 98474 AVERAGE: STANDARD DEVIATION: RELATIVE STANDARD DEVIATION: % Recovery 89 91 94 92 107 105 101 99 59 61 99 99 91 16 17 * Duplicate Injection LOQ = 10 ppb C entre A nalytical Laboratories, Inc. Page 25 o f 102 000267 C entre Study N o.: 023-063 Table VII. Summary of PFOS Recoveries in Mallard Yolk Sponsor m Na Na Na Na Na Na Na Na Centre ID 0003799 Spk A 0003799 Spk A* 0003799 Spk B 0003799 Spk B* 0003799 Spk A 0003799 Spk A* 0003799 Spk B 0003799 Spk B* Set Number 032601B 032601B 032601B 032601B 032701A 032701A 032701A 032701A Extraction Analysis Amt Date Date Added (ppb) 3/26/01 3/26/01 3/26/01 3/26/01 3/27/01 3/27/01 3/27/01 3/27/01 10 10 98 98 3/27/01 3/27/01 3/27/01 3/27/01 3/28/01 3/28/01 3/28/01 3/28/01 10 10 97 97 AVERAGE: STANDARD DEVIATION: RELATIVE STANDARD DEVIATION: % Recovery 104 104 99 99 71 73 98 94 93 13 14 Table VIII. Summary of PFOS for Week 1 Mallard Membrane Control Samples Sponsor ID E01-0263-20852 E01-0263-20852* E01-0263-20853 E01-0263-20853* E01-0263-20854 E01-0263-20854* EO1-0263-20842 E01-0263-20842* E01-0263-20843 E01-Q263-20843* Centre ID 0103200 0103200 0103204 0103204 0103208 0103208 0103160 0103160 0103164 0103164 * Duplicate Injection Set Number 032601C 032601C 032601C 032601C 032601C 032601C 032601C 032601C 032601C 032601C Extraction Analysis Analyte Date Date Found(ppb) 3/26/01 3/26/01 3/27/01 3/27/01 NQ NQ 3/26/01 3/26/01 3/27/01 3/27/01 NQ NQ 3/26/01 3/26/01 3/26/01 3/27/01 3/27/01 3/27/01 NQ NQ NQ 3/26/01 3/27/01 NQ 3/26/01 3/26/01 3/27/01 NQ 3/27/01 ______NQ____ AVERAGE: NQ STANDARD DEVIATION: NQ NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/tnL)) LOQ = 10 ppb C entre A nalytical Laboratories, Inc. Page 26 o f 102 000268 C entre Study N o.: 023-063 Table IX. Summary of PFOS for Week 1 Mallard Membrane Samples at 6.2 ppm ai Sponsor ID EOI-0263-20825 EOI-0263-20825* EOI-0263-20846 EOI-0263-20846* E01-Q263-20832 EOl-0263-20832* E01-0263-20847 E01-0263-20847* E01-0263-20848 E01-0263-20848* Centre ID 0103092 0103092 0103176 0103176 0103120 0103120 0103180 0103180 0103184 0103184 Set Number 032601C 032601C 032601C 032601C 032601C 032601C 032601C 032601C 032601C 032601C Extraction Date Analysis Date 3/26/01 3/27/01 3/26/01 3/27/01 3/26/01 3/27/01 3/26/01 3/27/01 3/26/01 3/27/01 3/26/01 3/27/01 3/26/01 3/27/01 3/26/01 3/27/01 3/26/01 3/27/01 3/26/01 3/27/01 AVERAGE: STANDARD DEVIATION: Analyte Found (ppb) 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 at Sponsor ro EOI-0263-20856 EOI-0263-20856* E01-0263-20850 E01-0263-20850* E01-0263-20857 E01-0263-208S7* E01-0263-20851 E01-0263-20851* E01-0263-20858 E01-0263-20858* Centre ED 0103216 0103216 0103192 0103192 0103220 0103220 01031% 0103196 0103224 0103224 Set Number 032601C 032601C 032601C 032601C 032601C 032601C 032601C 032601C 032601C 032601C Extraction Analysis Analyte Date Date Found(ppb) 3/26/01 3/27/01 3/26/01 3/27/01 3/26/01 3/27/01 3/26/01 3/27/01 3/26/01 3/27/01 3/26/01 3/27/01 3/26/01 3/27/01 3/26/01 3/27/01 3/26/01 3/27/01 3/26/01 3/27/01 AVERAGE: STANDARD DEVIATION: 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. C entre A nalytical Laboratories, Inc. Page 27 o f 102 C entre Study N o.: 023-063 Table XI. Summary of PFOS for Week I Mallard Albumen Control Samples Sponsor ID E01-0263-20852 E01-Q263-20852* E01-0263-20853 E01-0263-20853* E0I-0263-20854 E01-0263-20854* E01-0263-20842 EOI-0263-20842* EOl-0263-20843 E01-Q263-20843* Centre ID 0103201 0103201 0103205 0103205 0103209 0103209 0103161 0103161 0103165 0103165 Set Number 032601A 032601A 032601A 032601A 032601A 032601A 032601A 032601A 032601A 032601A Extraction Analysis Analyte Date Date Found(ppb) 3/26/01 3/26/01 3/26/01 3/26-27/01 3/26-27/01 3/26-27/01 NQ NQ NQ 3/26/01 3/26-27/01 NQ 3/26/01 3/26/01 3/26/01 3/26/01 3/26-27/01 3/26-27/01 3/26-27/01 3/26-27/01 NQ NQ NQ NQ 3/26/01 3/26-27/01 NQ 3/26/01 3/26-27/01 _____ NQ___ AVERAGE: NQ STANDARD DEVIATION: NQ Table XII. Summary of PFOS for Week I Mallard Albumen Samples at 6.2 ppm ai Sponsor ID E01-0263-20825 E01-Q263-20825* E01-0263-20846 E01-0263-20846* E01-0263-20832 E 0 1-0263-20832* E01-0263-20847 E01-0263-20847* E01-0263-20848 E01-0263-20848* Centre ID 0103093 0103093 0103177 0103177 0103121 0103121 0103181 0103181 0103185 0103185 Set Number 032601A 032601A 032601A 032601A 032601A 032601A 032601A 032601A 032601A 032601A Extraction Analysis Analyte Date Date Found(ppb) 3/26/01 3/26/01 3/26/01 3/26/01 3/26/01 3/26/01 3/26/01 3/26/01 3/26/01 3/26/01 3/26-27/01 3/26-27/01 3/26-27/01 3/26-27/01 3/26-27/01 3/26-27/01 3/26-27/01 3/26-27/01 3/26-27/01 3/26-27/01 2.49 2.75 7.90 7.89 5.67 5.79 5.76 5.58 5.95 6.29 AVERAGE: STANDARD DEVIATION: 5.63 1.80 * Duplicate Injection NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) LOQ = 10 ppb C entre A nalytical Laboratories, Inc. Page 28 o f 102 C entre Study No.: 023-063 Table XIII. Summary of PFOS for Week 1 Mallard Albumen Samples at 17.6 ppm ai Sponsor ID E01-0263-20856 E01-0263-20856* E01-0263-20856 E01-0263-20856* E01-0263-20856 EQ1-0263-20856* E01-0263-20850 E01-0263-20850* E01-0263-20857 E01-0263-20857* E01-0263-20851 E01-0263-20851* E01-0263-20858 E01-0263-20858* Centre ID 0103217 0103217 0103217 0103217 0103217Dup 0103217Dup 0103193 0103193 0103221 0103221 0103197 0103197 0103225 0103225 Set Number 032601D 032601D 033001C 033001C 033001C 033001C 032601A 032601A 032601A 032601A 032601A 032601A 032601A 032601A Extraction Analysis Analyte Date Date Found(ppb) 3/26/01 3/26/01 3/30/01 3/30/01 3/30/01 3/27-28/01 3/27-28/01 4/2/01 4/2/01 4/2/01 3550** 3490** 1388** 1369** 845** 3/30/01 4/2/01 771** 3/26/01 3/26/01 3/26-27/01 3/26-27/01 24.7 23.9 3/26/01 3/26/01 3/26/01 3/26/01 3/26/01 3/26/01 3/26-27/01 3/26-27/01 3/26-27/01 3/26-27/01 3/26-27/01 3/26-27/01 23.8 23.3 78.0** 69.7** 19.7 19.8 AVERAGE: 22.5 STANDARD DEVIATION: 2.20 Table XTV. Summary of PFOS for Week 1 Mallard Yolk Control Samples Sponsor ID E01-0263-20852 E01-0263-20852* EOl-0263-20853 E01-0263-20853* E01-0263-20854 E01-0263-20854* E01-0263-20842 E01-0263-20842* E01-0263-20843 E01-Q263-20843* Centre ID 0103198 0103198 0103202 0103202 0103206 0103206 0103158 0103158 0103162 0103162 * Duplicate Injection Set . Number 032601B 032601B 0326O1B 032601B 032601B 032601B 032601B 032601B 03260IB 032601B Extraction Analysis Analyte Date Date Found(ppb) 3/26/01 3/27/01 3/26/01 3/27/01 3/26/01 3/27/01 3/26/01 3/27/01 3/26/01 3/27/01 3/26/01 3/27/01 3/26/01 3/27/01 3/26/01 3/27/01 3/26/01 3/27/01 3/26/01 3/27/01 AVERAGE: STANDARD DEVIATION: NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) LOQ= lOppb ** Possible contamination by yolk or outer shell component of sample and as per Sponsor Representative request were not used to calculate the average and standard deviation. Centre Analytical Laboratories, Inc. Page 29 of 102 000271 C entre Study N o.: 023-063 Table XV. Summary of PFOS for Week 1 Mallard Yolk Samples at 6.2 ppmai Sponsor ID E01-0263-20825 E01-0263-20825* E01-0263-20846 E01-0263-20846* E01-0263-20832 E01-0263-20832* E01-0263-20847 E01-0263-20847* E01-0263-20848 E01-0263-20848* Centre ID 0103090 0103090 0103174 0103174 0103118 0103118 0103178 0103178 0103182 0103182 Set Number 032601D 032601D 032601D 03260ID 032601D 032601D 032601D 032601D 032601D 032601D Extraction Date Analysis Date 3/26/01 3/27-28/01 3/26/01 3/27-28/01 3/26/01 3/27-28/01 3/26/01 3/27-28/01 3/26/01 3/27-28/01 3/26/01 3/27-28/01 3/26/01 3/27-28/01 3/26/01 3/27-28/01 3/26/01 3/27-28/01 3/26/01 3/27-28/01 AVERAGE: STANDARD DEVIATION: Analyte Found(ppb) 2470 2580 12500 12100 4010 3890 10500 11100 12100 12100 8340 4450 Table XVI. Summary of PFOS for Week 1 Mallard Yolk Samples at 17.6 ppm ai Sponsor ID E01-0263-20856 E01-0263-20856* E01-0263-20850 E01-0263-20850* E01-0263-20857 E01-0263-20857* E01-0263-20851 E01-0263-20851* E01-0263-20858 E01-0263-20858* Centre ID 0103214 0103214 0103190 0103190 0103218 0103218 0103194 0103194 0103222 0103222 * Duplicate Injection LOQ = 10 ppb Set Number 032601D 032601D 032601D 032601D 032601D 032601D 032601D 032601D 032601D 032601D Extraction Analysis Date Date 3/26/01 3/27-28/01 3/26/01 3/27-28/01 3/26/01 3/27-28/01 3/26/01 3/27-28/01 3/26/01 3/27-28/01 3/26/01 3/27-28/01 3/26/01 3/27-28/01 3/26/01 3/27-28/01 3/26/01 3/27-28/01 3/26/01 3/27-28/01 AVERAGE: STANDARD DEVIATION: Analyte Found (ppb) 30300 30000 40000 43000 57200 56600 51700 53100 61000 62000 48500 12000 C entre A nalytical Laboratories, Inc. Page 30 o f 102 C entre Study N o.: 023-063 Table XVII. Summary of PFOS for Week 6 Mallard Membrane Control Samples Sponsor ID E01-0263-20993 E01-0263-20993* E01-0263-20994 E01-0263-20994* E01-0263-20995 E01-Q263-20995* E01-0263-20975 E01-0263-20975* E01-0263-20997 EOI-0263-20997* Centre ID 0103764 0103764 0103768 0103768 0103772 0103772 0103692 0103692 0103780 0103780 Set Number 032701C 032701C 032701C 032701C 032701C 032701C 032701C 032701C 032701C 032701C Extraction Analysis Date Date 3/27/01 3/27/01 3/29/01 3/29/01 3/27/01 3/29/01 3/27/01 3/29/01 3/27/01 3/27/01 3/29/01 3/29/01 3/27/01 3/27/01 3/27/01 3/29/01 3/29/01 3/29/01 3/27/01 3/29/01 AVERAGE: STANDARD DEVIATION: Analyte Found (nob) NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ Table XVIII. Summary of PFOS for Week 6 Mallard Membrane Samples at 6.2 ppm ai Sponsor ID E01-0263-20979 E01-0263-20979* E01-0263-21001 E01-0263-21001* E01-0263-20988 E01-0263-20988* E01-0263-20971 E01-0263-20971* E01-0263-20989 E01-0263-20989* Centre ID 0103708 0103708 0103796 0103796 0103744 0103744 0103676 0103676 0103748 0103748 * Duplicate Injection Set Number 032701C 032701C 032701C 032701C 032701C 032701C 032701C 032701C 032701C 032701C Extraction Analysis Analyte Date Date Found (ppb) 3/27/01 3/29/01 3/27/01 3/29/01 3/27/01 3/29/01 3/27/01 3/29/01 3/27/01 3/29/01 3/27/01 3/29/01 3/27/01 3/29/01 3/27/01 3/29/01 3/27/01 3/29/01 3/27/01 3/29/01 AVERAGE: STANDARD DEVIATION: 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. C entre A nalytical L aboratories, Inc. Page 31 o f 102 000273 C entre Study No.: 023-063 Table XIX. Summary of PFOS for Week 6 Mallard Membrane Samples at 17.6 ppm ai Sponsor ID EOI-0263-20991 E01-0263-20991* EOI-0263-20990 E01-0263-20990* E01-0263-21002 E01-0263-21002* E01-0263-21003 EOI-0263-21003* E01-0263-21004 EOI-0263-21004* Centre ID 0103756 0103756 0103752 0103752 0103800 0103800 0103804 0103804 0103808 0103808 Set Number 032701CD 032701CD 032701CD 032701CD 032701CD 032701CD 032701CD 032701CD 032701CD 032701CD Extraction Analysis Date Date 3/27/01 3/27/01 3/29/01 3/29/01 3/27/01 3/29/01 3/27/01 3/29/01 3/27/01 3/27/01 3/27/01 3/27/01 3/27/01 3/27/01 3/29/01 3/29/01 3/29/01 3/29/01 3/29/01 3/29/01 AVERAGE: STANDARD DEVIATION: Analyte Found (ppb) 434 431 365 370 347 339 306 307 259 261 342 61.2 Table XX. Summary of PFOS for Week 6 Mallard Albumen Control Samples Sponsor ID E01-0263-20993 E01-0263-20993* E01-0263-20994 E01-0263-20994* E01-0263-20995 E01-0263-20995* E01-0263-20975 E01-0263-20975* E01-0263-20997 E01-0263-20997* Centre ED 0103765 0103765 0103769 0103769 0103773 0103773 0103693 0103693 0103781 0103781 * Duplicate Injection Set Number 032701B 032701B 032701B 032701B 032701B 032701B 032701B 03270IB 032701B 032701B Extraction Analysis Analyte Date Date Found(ppb) 3/27/01 3/27/01 3/27/01 3/27/01 3/27/01 3/27/01 3/27/D1 3/28-29/01 3/28-29/01 3/28-29/01 3/28-29/01 3/28-29/01 3/28-29/01 3/28-29/D1 NQ NQ NQ NQ NQ NQ NQ 3/27/01 3/28-29/01 3/27/01 3/28-29/01 3/27/01 3/28-29/01 AVERAGE: STANDARD DEVIATION: NQ NQ NQ NQ NQ NQ ss Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) LOQ = 10 ppb C entre A nalytical Laboratories, Inc. Page 32 o f 102 000274 C entre Study No.: 023-063 Table XXI. Summary of PFOS for Week 6 Mallard Albumen Samples at 6.2 ppm ai Sponsor ID E01-0263-20979 E01-0263-20979* E01-0263-21001 E01-Q263-21001* E01-0263-20988 E01-0263-20988* E01-0263-20971 E01-0263-20971* E01-0263-20989 E01-0263-20989* Centre ID 0103709 0103709 0103797 0103797 0103745 0103745 0103677 0103677 0103749 0103749 Set Number 032701B 032701B 032701B 032701B 032701B 032701B 032701B 032701B 032701B 032701B Extraction Analysis Date Date 3/27/01 3/28-29/01 3/27/01 3/28-29/01 3/27/01 3/28-29/01 3/27/01 3/28-29/01 3/27/01 3/28-29/01 3/27/01 3/28-29/01 3/27/01 3/28-29/01 3/27/01 3/28-29/01 3/27/01 3/28-29/01 3/27/01 3/28-29/01 AVERAGE: STANDARD DEVIATION: Analyte Found(ppb) 5.83 6.23 7.92 8.04 6.56 6.26 7.02 6.81 8.72 8.33 7.19 0.987 Table XXII. Summary of PFOS for Week 6 Mallard Albumen Samples at 17.6 ppm ai Sponsor ID E01-0263-20991 E01-0263-20991* E01-0263-20990 E01-0263-20990* E01-0263-21002 E01-0263-21002* E01-0263-21003 E01-0263-21003* E01-0263-21004 E01-0263-21004* Centre ID 0103757 0103757 0103753 0103753 0103801 0103801 0103805 0103805 0103809 0103809 * Duplicate Injection LOQ = 10 ppb Set Number 032701B 032701B 032701B 03270IB 032701B 032701B 032701B 03270IB 032701B 032701B Extraction Analysis Date Date 3/27/01 3/28-29/01 3/27/01 3/27/01 3/27/01 3/27/01 3/27/01 3/28-29/01 3/28-29/01 3/28-29/01 3/28-29/01 3/28-29/01 3/27/01 3/27/01 3/27/01 3/27/01 3/28-29/01 3/28-29/01 3/28-29/01 3/28-29/01 AVERAGE: STANDARD DEVIATION: Analyte Found(ppb) 25.3 24.9 14.1 14.7 19.7 19.4 23.6 24.0 19.1 17.2 20.2 4.12 C entre A nalytical L aboratories, Inc. Page 33 o f 102 000275 C entre Study No.: 023-063 Table XXIII. Summary of PFOS for Week 6 Mallard Yolk Control Samples Sponsor m E0I-O263-20993 E01-0263-20993* E0I-0263-20994 E01-0263-20994* E01-0263-20995 E01-0263-20995* E01-0263-20975 E01-0263-20975* E01-0263-20997 E01-0263-20997* Centre ID 0103762 0103762 0103766 0103766 0103770 0103770 0103690 0103690 0103778 0103778 Set Number 032701A 032701A 032701A 032701A 032701A 032701A 032701A 032701A 032701A 032701A Extraction Analysis Date Date 3/27/01 3/28/01 3/27/01 3/28/01 3/27/01 3/28/01 3/27/01 3/28/01 3/27/01 3/28/01 3/27/01 3/28/01 3/27/01 3/28/01 3/27/01 3/28/01 3/27/01 3/28/01 3/27/01 3/28/01 AVERAGE: STANDARD DEVIATION: Analyte Found(ppb) NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ Table XXIV. Summary of PFOS for Week 6 Mallard Yolk Samples at 6.2 ppm ai Sponsor ID E01-0263-20979 E01-0263-20979* E01-0263-21001 E01-0263-21001* E01-0263-20988 EOI-0263-20988* EO1-0263-20971 E01-0263-20971* E01-0263-20989 E01-0263-20989* Centre ID 0103706 0103706 0103794 0103794 0103742 0103742 0103674 0103674 0103746 0103746 * Duplicate Injection Set Number 032701AD 032701AD 032701AD 032701AD 032701AD 032701AD 032701AD 032701AD 032701AD 032701AD Extraction Analysis Date Date 3/27/01 3/27/01 3/27/01 3/27/01 3/27/01 3/27/01 3/27/01 3/27/01 3/27/01 3/27/01 3/29/01 3/29/01 3/29/01 3/29/01 3/29/01 3/29/01 3/29/01 3/29/01 3/29/01 3/29/01 AVERAGE: STANDARD DEVIATION: Analyte Found(ppb) 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 C entre A nalytical Laboratories, Inc. Page 34 o f 102 C entre Study N o.: 023-063 Table XXV. Summary of PFOS for Week 6 Mallard Yolk Samples at 17.6 ppm ai Sponsor ID E01-0263-20991 E01-0263-20991* EQ1-0263-20990 E01-0263-20990* E01-0263-21002 E01-0263-21002* E01-0263-21003 E01-0263-21003* E01-0263-21004 E01-0263-21004* Centre ID 0103754 0103754 0103750 0103750 0103798 0103798 0103802 0103802 0103806 0103806 Set Number 032701AD 032701AD 032701AD 032701AD 032701AD 032701AD 032701AD 032701AD 032701AD 032701AD Extraction Analysis Date Date 3/27/01 3/29/01 3/27/01 3/29/01 3/27/01 3/29/01 3/27/01 3/29/01 3/27/01 3/29/01 3/27/01 3/29/01 3/27/01 3/29/01 3/27/01 3/29/01 3/27/01 3/29/01 3/27/01 3/29/01 AVERAGE: STANDARD DEVIATION: Analyte Found(ppb) 41900 40700 63000 63500 57500 57300 43100 43600 46600 49800 50700 8870 Table XXVI. Summary of PFOS in Quail Membrane Blanks Sponsor ID na na na na Centre ID 0003798 Blank A 0003798 Blank A* 0003798 Blank A 0003798 Blank A* Set Extraction Analysis Number Date Date 032901A 3/29/01 3/31/01 032901A 033001BR 033001BR 3/29/01 3/30/01 3/30/01 3/31/01 4/2/01 4/2/01 AVERAGE: STANDARD DEVIATION: Analyte Found (ppb) NQ NQ NQ NO NQ NQ Table XXVII. Summary of PFOS in Quail Albumen Blanks Sponsor ID na na na na Centre ID 0003796 Blank A 0003796 Blank A* 0003796 Blank A 0003796 Blank A* * Duplicate Injection Set Number 032801B 032801B 033001A 033001A Extraction Analysis Date Date 3/28/01 3/28/01 3/30/01 3/30/01 3/30/01 3/30/01 4/1/01 4/1/01 AVERAGE: STANDARD DEVIATION: Analyte Found(ppb) NQ NQ NQ NO NQ NQ NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) LOQ = 10 ppb C entre A nalytical L aboratories, Inc. Page 35 o f 102 C entre Study No.: 023-063 Table XXVIII. Summary of PFOS in Quail Yolk Blanks Sponsor ID na na na na Centre ID 0003795 Blank A 0003795 Blank A* 0003795 Blank A 0003795 Blank A* Set Number 032801A 032801A 032901B 032901B Extraction Analysis Date Date 3/28/01 3/28/01 3/29/01 3/29/01 3/30/01 3/30/01 3/31-4/1/01 3/31-4/1/01 AVERAGE: STANDARD DEVIATION: Analyte Found(ppb) NQ NQ ND ND NQ NQ Table XXIX. Summary of PFOS Recoveries in Quail Membrane Sponsor ID na na na na na na na na Centre ID 0003798 Spk A 0003798 Spk A* 0003798 Spk B 0003798 Spk B* 0003798 Spk A 0003798 Spk A* 0003798 Spk B 0003798 Spk B* Set Number Q32901A 032901A 032901A 032901A 033001BR 033001BR 033001B 033001B Extraction Analysis Amt Date Date Added (ppb) 3/29/01 3/29/01 3/31/01 3/31/01 10 10 3/29/01 3/31/01 986 3/29/01 3/31/01 986 3/30/01 4/2/01 10 3/30/01 4/2/01 10 3/30/01 4/2/01 98 3/30/01 4/2/01 98 AVERAGE: STANDARD DEVIATION: RELATIVE STANDARD DEVIATION: % Recovery 102 104 88 85 114 116 108 101 102 II 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 m C entre A nalytical L aboratories, Inc. Page 36 o f 102 000278 C entre Study N o.: 023-063 Table XXX. Summary of PFOS Recoveries in Quail Albumen Sponsor ID na na na na na na na na Centre ID 0003796 Spk A 0003796 Spk A* 0003796 Spk B 0003796 Spk B* 00037% Spk A 0003796 Spk A* 0003796 Spk B 0003796 Spk B* Set Number 032801B 032801B 032801B 032801B 033001A 033001A 033001A 033001A Extraction Analysis Amt Date Date Added (nob) 3/28/01 3/28/01 3/28/01 3/28/01 3/30/01 3/30/01 3/30/01 3/30/01 3/30/01 3/30/01 3/30/01 3/30/01 4/1/01 4/1/01 4/1/01 4/1/01 10 10 9963 9963 10 10 99 99 AVERAGE: STANDARD DEVIATION: RELATIVE STANDARD DEVIATION: % Recovery 82 79 80 83 82 88 93 91 85 5 6 Table XXXI. Summary of PFOS Recoveries in Quail Yolk Sponsor ID na na na na na na na na Centre ID 0003795 Spk A 0003795 Spk A* 0003795 Spk B 0003795 Spk B* 0003795 Spk A 0003795 Spk A* 0003795 Spk B 0003795 Spk B* Set Number 032801A 032801A 032801A 032801A 03290IB 032901B 032901D 032901D Extraction Analysis Amt Date Date Added (ppb) 3/28/01 3/30/01 10 3/28/01 3/30/01 10 3/28/01 3/30/01 %23 3/28/01 3/30/01 9623 3/29/01 3/31-4/1/01 10 3/29/01 3/31-4/1/01 10 3/29/01 4/1-2/01 98834 3/29/01 4/1-2/01 98834 AVERAGE: STANDARD DEVIATION: RELATIVE STANDARD DEVIATION: % Recovery 102 104 98 101 87 87 94 93 96 7 7 * Duplicate Injection LOQ = 10 ppb C entre A nalytical Laboratories, Inc. Page 37 o f 102 000279 Centre Study No.: 023-063 Table XXXII. Summary of PFOS for Week 1 Quail Membrane Control Samples Sponsor Centre Set Extraction Analysis Analyte ID ID Number Date Date Found(ppb) E01-0259-20506 E01-0259-20506* E01-0259-20507 0101987 0101987 0101991 032901A 032901A 032901A 3/29/01 3/29/01 3/29/01 3/31/01 3/31/01 3/31/01 5.20** 5.57** NQ EOI-0259-20507* E01-0259-20520 0101991 0102043 032901A 032901A 3/29/01 3/29/01 3/31/01 3/31/01 NQ NQ E01-0259-20520* E01-0259-20521 E01-0259-20521* E01-0259-20509 EQI-0259-20509* 0102043 0102047 0102047 0101999 0101999 032901A 032901A 032901A 032901A 032901A 3/29/01 3/31/01 3/29/01 3/31/01 3/29/01 3/31/01 3/29/01 3/31/01 3/29/01 3/31/01 AVERAGE: STANDARD DEVIATION: NQ NQ NQ NQ NO NQ NQ Table XXXHI. Summary of PFOS for Week 1 Quail Membrane Samples at 6.2 ppm ai Sponsor Centre Set Extraction Analysis Analyte ID m Number Date Date Found (ppb) E01-0259-2Q526 E01-0259-20526* E01-0259-2Q514 E01-0259-20514* 0102067 0102067 0102019 0102019 032901D 032901D 032901A 032901A 3/29/01 3/29/01 3/29/01 3/29/01 4/1-2/01 4/1-2/01 3/31/01 3/31/01 18800** 17900** 127 129 E01-0259-20527 0102071 032901A 3/29/01 3/31/01 123 E01-0259-20527* E01-0259-20515 E01-0259-20515* E01-0259-20528 E01-0259-20528* 0102071 0102023 0102023 0102075 0102075 032901A 032901A 032901A 032901D 032901D 3/29/01 3/29/01 3/29/01 3/29/01 3/29/01 3/31/01 3/31/01 3/31/01 4/1-2/01 4/1-2/01 116 172 172 11500** 11500** * Duplicate Injection AVERAGE: STANDARD DEVIATION: 140 253 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 C entre Study N o.: 023-063 Table XXXIV. Summary of PFOS for Week 1 Quail Membrane Samples at 17.6 ppm ai Sponsor ID E01-0259-20529 E01-0259-20529* E01-0259-20531 E01-0259-20531* E01-0259-20532 E01-0259-20532* E01-0259-20518 E01-0259-20518* E01-0259-20533 E01-0259-20533* Centre m 0102079 0102079 0102087 0102087 0102091 0102091 0102035 0102035 0102095 0102095 Set Number 032901D 032901D 032901D 032901D 032901D 032901D 032901D 032901D 032901D 032901D Extraction Analysis Analyte Date Date Found(ppb) 3/29/01 4/1-2/01 3/29/01 4/1-2/01 3/29/01 4/1-2/01 3/29/01 4/1-2/01 3/29/01 4/1-2/01 3/29/01 4/1-2/01 3/29/01 4/1-2/01 3/29/01 4/1-2/01 3/29/01 4/1-2/01 3/29/01 4/1-2/01 AVERAGE: STANDARD DEVIATION: 20900 20400 11000 11100 14300 13900 12500 12600 6880 6990 13100 4730 Table XXXV. Summary of PFOS for Week 1 Quail Albumen Control Samples Sponsor ID E01-0259-20506 E01-0259-20506* E01-0259-20507 E01-0259-20507* E01-0259-20520 E01-Q259-20520* E01-0259-20521 E01-0259-20521* E01-0259-20509 E01-0259-20509* Centre ID 0101988 0101988 0101992 0101992 0102044 0102044 0102048 0102048 0102000 0102000 Set Number 032801B 032801B 032801B 03280IB 032801B 032801B 03280IB 03280IB 032801B 032801B Extraction Analysis Analyte Date Date Found(ppb) 3/28/01 3/30/01 NQ 3/28/01 3/30/01 NQ 3/28/01 3/28/01 3/28/01 3/28/01 3/28/01 3/28/01 3/28/01 3/28/01 3/30/01 NQ 3/30/01 NQ 3/30/01 NQ 3/30/01 NQ 3/30/01 NQ 3/30/01 NQ 3/30/01 NQ 3/30/01 ______m ___ AVERAGE: NQ STANDARD DEVIATION: NQ * Duplicate Injection NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) LOQ = 10 ppb C entre A nalytical L aboratories. Inc. Page 39 o f 102 C entre Study N o.: 023-063 Table XXXVI. Summary of PFOS for Week 1 Quail Albumen Samples at 6.2 ppm ai Sponsor ID E01-0259-20526 E01-0259-20526* E01-0259-20514 E01-0259-20514* E01-0259-20527 EOI-0259-20527* EOl-0259-20515 E01-0259-20515* E01-0259-20528 E01-0259-20528* Centre ID 0102068 0102068 0102020 0102020 0102072 0102072 0102024 0102024 0102076 0102076 Set Number 032801B 032801B 03280IB 032801B 032801B 032801B 03280IB 032801B 032801B 032801B Extraction Date Analysis Date Analyte Found (ppb) 3/28/01 3/30-31/01 3/28/01 3/30-31/01 3/28/01 3/30-31/01 3/28/01 3/30-31/01 3/28/01 3/30-31/01 3/28/01 3/30-31/01 3/28/01 3/30-31/01 3/28/01 3/30-31/01 3/28/01 3/30-31/01 3/28/01 3/30-31/01 AVERAGE: STANDARD DEVIATION: 6.03 5.43 2.43 2.78 2.94 3.48 5.87 6.08 93.9** 93.7** 438 1.61 Table XXXVII. Summary of PFOS for Week 1 Quail Albumen Samples at 17.6 ppm ai Sponsor ID E01-0259-20529 E01-0259-20529* E01-0259-2053I E01-0259-20531* E0I-0259-20532 E01-0259-20532* EOI-0259-20518 E01-0259-20518* E01-0259-20533 E01-0259-20533* Centre ID 0102080 0102080 0102088 0102088 0102092 0102092 0102036 0102036 0102096 0102096 * Duplicate Injection LOQ = 10 ppb Set Number 032801B 032801B 032801B 03280IB 032801B 032801B 032801B 032801B 032801B 032801B Extraction Analysis Analyte Date Date Found (ppb) 3/28/01 3/30-31/01 3/28/01 3/30-31/01 3/28/01 3/30-31/01 3/28/01 3/30-31/01 3/28/01 3/30-31/01 3/28/01 3/30-31/01 3/28/01 3/30-31/01 3/28/01 3/30-31/01 3/28/01 3/30-31/01 3/28/01 3/30-31/01 AVERAGE: STANDARD DEVIATION: 17.9 17.3 9.97 10.5 10.7 11.0 32.3 31.6 9.07 10.1 16.0 8.92 C entre A nalytical L aboratories. Inc. Page 40 o f 102 C entre Study No.: 023-063 Table XXXVIII. Summary of PFOS for Week 1 Quail Yolk Control Samples Sponsor ID E01-0259-20506 E01-0259-20506* E01-0259-20507 E01-0259-20507* E01-0259-20520 E01-0259-20520* E01-0259-20521 E01-0259-20521* E01-0259-20509 E01-0259-20509* Centre ID 0101985 0101985 0101989 0101989 0102041 0102041 0102045 0102045 0101997 0101997 Set Number 032801A 032801A 032801A 032801A 032801A 032801A 032801A 032801A 032801A 032801A Extraction Analysis Analyte Date Date Found (unb) 3/28/01 3/30/01 3/28/01 3/30/01 3/28/01 3/30/01 3/28/01 3/30/01 3/28/01 3/30/01 3/28/01 3/30/01 3/28/01 3/30/01 3/28/01 3/30/01 3/28/01 3/30/01 3/28/01 3/30/01 AVERAGE: STANDARD DEVIATION: NQ NQ NQ NQ NQ NQ NQ NQ NQ NO NQ NQ Table XXXIX. Summary of PFOS for Week 1 Quail Yolk Samples at 6.2 ppmai Sponsor ID E01-0259-20526 E01-0259-20526* E01-0259-2Q514 E01-0259-20514* E01-0259-20527 E01-0259-20527* E01-0259-20515 E01-0259-20515* E01-0259-20528 E01-0259-20528* Centre ID 0102065 0102065 0102017 0102017 0102069 012069 0102021 0102021 0102073 0102073 * Duplicate Injection Set Number 032801AD 032801AD 032801AD 032801AD 032801AD 032801AD 032801AD 032801AD 032801AD 032801AD Extraction Analysis Date Date 3/28/01 3/28/01 3/31/01 3/31/01 3/28/01 3/28/01 3/31/01 3/31/01 3/28/01 3/31/01 3/28/01 3/28/01 3/31/01 3/31/01 3/28/01 3/28/01 3/31/01 3/31/01 3/28/01 3/31/01 AVERAGE: STANDARD DEVIATION: Analyte Found(ppb) 17500 17500 8530 8410 12100 12200 13800 13800 11700 10900 12600 3150 NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) LOQ = 10 ppb C entre A nalytical L aboratories, Inc. Page 41 o f 102 000283 C entre Study No.: 023-063 Table XL. Summary of PFOS for Week 1 Quail Yolk Samples at 17.6 ppm ai Sponsor Centre Set Extraction Analysis Analyte ID ID Number Date Date Found(ppb) E01-0259-20529 E01-0259-20529* 0102077 0102077 032801AD 032801AD 3/28/01 3/28/01 3/31/01 3/31/01 42500 41200 E01-0259-20531 E01-0259-20531* E01-0259-20532 E01-0259-20532* E01-0259-20518 E01-0259-20518* E01-0259-20533 E01-0259-20533* 0102085 0102085 0102089 0102089 0102033 0102033 0102093 0102093 032801AD 032801AD 032801AD 032801AD 032801AD 032801AD 032801AD 032801AD 3/28/01 3/28/01 3/28/01 3/28/01 3/28/01 3/28/01 3/28/01 3/28/01 3/31/01 3/31/01 3/31/01 3/31/01 3/31/01 3/31/01 3/31/01 3/31/01 33200 31100 37400 38900 38100 38300 43000 43600 AVERAGE: STANDARD DEVIATION: 38700 4130 Table XLI. Summary of PFOS for Week 6 Quail Membrane Control Samples Sponsor Centre Set Extraction Analysis Analyte ID ID Number Date Date Found(ppb) E01-0259-20735 E01-0259-20735* E01-0259-20736 E01-0259-20736* E01-0259-20737 E01-0259-20737* E01-0259-20720 E01-0259-20720* E01-0259-20738 E01-0259-20738* 0102903 0102903 0102907 0102907 0102911 0102911 0102843 0102843 0102915 0102915 * Duplicate Injection 033001B 033001B 03300IB 033001B 033001B 033001B 033001B 033001B 033001B 033001B 3/30/01 4/2/01 3/30/01 4/2/01 3/30/01 4/2/01 3/30/01 4/2/01 3/30/01 4/2/01 3/30/01 4/2/01 3/30/01 4/2/01 3/30/01 4/2/01 3/30/01 4/2/01 3/30/01 4/2/01 AVERAGE: STANDARD DEVIATION: NQ NQ 9.85** 8.93** 15.6** 14.4** 21.8** 24.4** 6.51** 6.30** NQ NQ NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) LOQ = lOppb ** 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. C entre A nalytical Laboratories, Inc. Page 42 o f 102 000284 000285 C entre Study No.: 023-063 Table XLTV. Summary of PFOS for Week 6 Quail Albumen Control Samples Sponsor ID E0I-0259-20735 E01-0259-20735* E01-Q259-20736 E01-0259-20736* E01-0259-20737 E01-0259-20737* E01-0259-20720 E01-0259-20720* E01-0259-20738 E01-0259-20738* Centre ID 0102904 0102904 0102908 0102908 0102912 0102912 0102844 0102844 0102916 0102916 Set Number 033001A 033001A 033001A 033001A 033001A 033001A 033001A 033001A 033001A 033001A Extraction Analysis Date Date 3/30/01 3/30/01 3/30/01 4/1/01 4/1/01 4/1/01 3/30/01 3/30/01 3/30/01 3/30/01 3/30/01 3/30/01 3/30/01 4/1/01 4/1/01 4/1/01 4/1/01 4/1/01 4/1/01 4/1/01 AVERAGE: STANDARD DEVIATION: Analyte Found(ppb) NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ Table XLV. Summary of PFOS for Week 6 Quail Albumen Samples at 6.2 ppm ai Sponsor ID E01-0259-20742 E01-Q259-20742* E01-0259-20743 E01-0259-20743* E01-0259-20744 E01-0259-20744* E01-0259-20745 E01-0259-20745* E01-0259-20746 E01-0259-20746* Centre ID 0102932 0102932 0102936 0102936 0102940 0102940 0102944 0102944 0102948 0102948 * Duplicate Injection Set Number 033001A 033001A 033001A 033001A 033001A 033001A 033001A 033001A 033001A 033001A Extraction Date Analysis Date 3/30/01 4/1/01 3/30/01 4/1/01 3/30/01 4/1/01 3/30/01 4/1/01 3/30/01 4/1/01 3/30/01 4/1/01 3/30/01 4/1/01 3/30/01 4/1/01 3/30/01 4/1/01 3/30/01 4/1/01 AVERAGE: STANDARD DEVIATION: Analyte Found(ppb) 5.34 5.93 6.19 6.91 6.82 6.82 5.26 5.15 3.56 4.34 5.63 1.12 NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) LOQ ss 10 ppb C entre A nalytical Laboratories, Inc. Page 44 o f 102 000286 C entre Study No.: 023-063 Table XLVI. Summary of PFOS for Week 6 Quail Albumen Samples at 17.6 ppm ai Sponsor ID Centre ID Set Number Extraction Date Analysis Date Analyte Found (ppb) E01-0259-20747 E01-Q259-20747* E01-0259-20748 E01-0259-20748* E01-0259-20749 E01-0259-20749* E01-0259-20750 E01-Q259-20750* E01-0259-20751 E01-0259-20751* 0102952 0102952 0102956 0102956 0102960 0102960 0102964 0102964 0102968 0102968 033001A 033001A 033001A 033001A 033001A 033001A 033001A 033001A 033001A 033001A 3/30/01 4/1/01 3/30/01 4/1/01 3/30/01 4/1/01 3/30/01 4/1/01 3/30/01 4/1/01 3/30/01 4/1/01 3/30/01 4/1/01 3/30/01 4/1/01 3/30/01 4/1/01 3/30/01 4/1/01 AVERAGE: STANDARD DEVIATION: 9.81 9.84 13.2 13.1 14.6 14.2 16.3 16.7 15.6 15.4 13.9 2.44 Table XLVII. Summary of PFOS for Week 6 Quail Yolk Control Samples Sponsor Centre Set Extraction Analysis Analyte ID ID Number Date Date Found (ppb) E01-0259-20735 E01-0259-20735* 0102901 0102901 032901B 032901B 3/29/01 3/29/01 3/31-4/1/01 3/31-4/1/01 NQ NQ E01-0259-20736 0102905 032901B 3/29/01 3/31-4/1/01 NQ E01-0259-20736* 0102905 03290IB 3/29/01 3/31-4/1/01 NQ E01-0259-20737 E01-0259-20737* 0102909 0102909 032901B 032901B 3/29/01 3/29/01 3/31-4/1/01 3/31-4/1/01 51.5** 51.1** E01-0259-20720 0102841 03290IB 3/29/01 3/31-4/1/01 NQ E01-0259-20720* E01-0259-20738 0102841 0102913 03290IB 032901B 3/29/01 3/29/01 3/31-4/1/01 3/31-4/1/01 NQ ND E01-0259-20738* 0102913 032901B 3/29/01 3/31-4/1/01 ND AVERAGE: NQ * Duplicate Injection STANDARD DEVIATION: NQ NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL ND = No peak detected LOQ = 10 ppb ** Possible contamination by outer shell component of sample since outer shell was not washed prior to separation of internal components. As per Sponsor Representative request these values were not used to calculate the average and standard deviation. C entre A nalytical L aboratories, Inc. Page 45 o f 102 C entre Study No.: 023-063 Table XLVIII. Summary of PFOS for Week 6 Quail Yolk Samples at 6.2 ppm ai Sponsor ID E01-0259-20742 E01-0259-20742* E01-0259-20743 E01-0259-20743* E01-0259-20744 E01-0259-20744* E01-0259-20745 E01-0259-20745* E01-0259-20746 E01-Q259-20746* Centre ID 0102929 0102929 0102933 0102933 0102937 0102937 0102941 0102941 0102945 0102945 Set Number 032901D 032901D 032901D 032901D 032901D 03290ID 03290ID 03290ID 03290ID 032901D Extraction Analysis Date Date 3/29/01 4/1-2/01 3/29/01 4/1-2/01 3/29/01 4/1-2/01 3/29/01 4/1-2/01 3/29/01 4/1-2/01 3/29/01 4/1-2/01 3/29/01 4/1-2/01 3/29/01 4/1-2/01 3/29/01 4/1-2/01 3/29/01 4/1-2/01 AVERAGE: STANDARD DEVIATION: Analyte Found (ppb) 38700\ 40500^ 37100^ 36KXT 23600. 236<KT 40000% 4060CK 24600v 25800^ 33100 7610 Table IL. Summary of PFOS for Week 6 Quail Yolk Samples at 17.6 ppm ai Sponsor ID E01-0259-20747 E01-Q259-20747* E01-0259-20748 E01-0259-20748* E01-0259-20749 E01-0259-20749* E01-0259-20750 E01-0259-20750* E01-0259-20751 E01-0259-20751* Centre ID 0102949 0102949 0102953 0102953 0102957 0102957 0102961 0102961 0102965 0102965 * Duplicate Iniection Set Number 032901D 03290ID 032901D 032901D 032901D 032901D 032901D 032901D 032901D 032901D Extraction Analysis Date Date 3/29/01 3/29/01 3/29/01 4/1-2/01 4/1-2/01 4/1-2/01 3/29/01 4/1-2/01 3/29/01 4/1-2/01 3/29/01 3/29/01 4/1-2/01 4/1-2/01 3/29/01 3/29/01 4/1-2/01 4/1-2/01 3/29/01 4/1-2/01 AVERAGE: STANDARD DEVIATION: Analyte Found(ppb) 53900v 54400-" 65500 64800^ 46400. 4650059400. 5930(r 56800 57800 56500 6510 NQ = Not Quantifiable (less than the concentration of the lowest standard (0.1 ng/mL)) C entre A nalytical L aboratories, Inc. Page 46 o f 102 000288 C entre Study No.: 023-063 Table L. Summary of Moisture Determinations for Mallard Membrane Samples Sponsor ID E01-0263-20852 E01-0263-20853 E01-0263-20854 E01-0263-20842 E01-0263-20843 E01-0263-20825 E01-0263-20846 E01-0263-20832 E01-0263-20847 E01-0263-20848 E01-0263-20856 E01-0263-20850 E01-0263-20857 E01-0263-20851 E01-0263-20858 E01-0263-20993 E01-0263-20994 EO1-0263-20995 E01-0263-20975 E01-0263-20997 E01-0263-20979 E01-0263-21001 E01-0263-20988 E01-0263-20971 E01-0263-20989 E01-0263-20991 EO1-0263-20990 E01-0263-21002 EO1-0263-21003 E01-Q263-21004 Centre ID 0103200 0103204 0103208 0103160 0103164 0103092 0103176 0103120 0103180 0103184 0103216 0103192 0103220 0103196 0103224 0103764 0103768 0103772 0103692 0103780 0103708 0103796 0103744 0103676 0103748 0103756 0103752 0103800 0103804 0103808 Average % Moisture 80.0 55.0 75.0 80.0 80.0 70.0 70.0 75.0 65.0 60.0 75.0 50.0 85.0 85.0 85.0 75.0 71.4 80.0 65.0 80.0 75.0 65.0 75.0 90.0 80.0 75.0 70.0 70.0 75.0 65.0 C entre A nalytical Laboratories, Inc. Page 47 o f 102 C entre Study N o.: 023-063 Table LI. Summary of Moisture Determinations for Mallard Albumen Samples Sponsor ID E01-0263-20852 E01-0263-20853 E01-0263-20854 E01-0263-20842 E01-0263-20843 E01-0263-20825 E01-0263-20846 E01-0263-20832 E01-0263-20847 E01-0263-20848 E01-0263-20856 E01-0263-20850 E01-0263-20857 E01-0263-20851 E01-0263-20858 EQ1-0263-20993 E01-0263-20994 EO1-0263-20995 E01-0263-20975 E01-0263-20997 E01-0263-20979 E01-0263-21001 E01-0263-20988 E01-0263-20971 E01-0263-20989 E01-0263-20991 E01-0263-20990 E01-0263-21002 E01-0263-21003 EO1-0263-21004 Centre ID 0103201 0103205 0103209 0103161 0103165 0103093 0103177 0103121 0103181 0103185 0103217 0103193 0103221 0103197 0103225 0103765 0103769 0103773 0103693 0103781 0103709 0103797 0103745 0103677 0103749 0103757 0103753 0103801 0103805 0103809 Average % Moisture 90.0 88.0 87.0 85.0 87.0 82.0 84.0 86.0 89.0 88.0 85.0 85.0 84.0 83.0 85.0 88.0 85.0 85.0 84.0 90.0 88.0 84.0 87.0 83.0 87.0 84.0 86.0 83.0 82.0 87.0 C entre A nalytical L aboratories, Inc. Page 48 o f 102 000290 C entre Study N o.: 023-063 Table LH. Summary of Moisture Determinations for Mallard Yolk Samples Sponsor ID E01-0263-20852 E01-0263-20853 E01-0263-20854 E01-0263-20842 E01-0263-20843 E01-0263-20825 E01-0263-20846 E01-0263-20832 E01-0263-20847 E01-0263-20848 E01-0263-20856 E01-0263-20850 E01-0263-20857 EO1-0263-20851 E01-0263-20858 E01-0263-20993 E01-0263-20994 E01-0263-20995 E01-0263-20975 E01-0263-20997 EO1-0263-20979 E01-0263-21001 E01-0263-20988 E01-0263-20971 E01-0263-20989 EO1-0263-20991 E01-0263-20990 EO1-0263-21002 E01-0263-21003 E01-0263-21004 Centre ID 0103198 0103202 0103206 0103158 0103162 0103090 0103174 0103118 0103178 0103182 0103214 0103190 0103218 0103194 0103222 0103762 0103766 0103770 0103690 0103778 0103706 0103794 0103742 0103674 0103746 0103754 0103750 0103798 0103802 0103806 Average % Moisture 44.0 45.0 48.0 45.0 43.0 54.0 49.0 43.0 43.0 48.0 45.0 50.0 49.0 48.0 45.0 45.0 44.0 59.0 45.0 46.0 45.0 48.0 45.0 48.0 50.0 56.0 42.0 51.0 44.9 44.0 C entre A nalytical Laboratories, Inc. Page 49 o f 102 000291 C entre Study N o.: 023-063 Table LIII. Summary of Moisture Determinations for Quail Membrane Samples Sponsor ID E01-0259-20507 E01-0259-2Q509 E01-0259-20526 E01-0259-20738 E01-0259-20742 E01-0259-20745 E01-0259-20748 E01-0259-20750 Centre ID 0101991 0101999 0102067 0102915 0102931 0102943 0102955 0102963 Average % Moisture 45.0 65.0 65.0 60.0 60.0 68.3 70.0 68.3 The percent moisture was detecmined for eight out of the 30 samples received because there was not enough sample available for the other samples. C entre A nalytical Laboratories, Inc. Page 50 of 102 600292 C entre Study N o.: 023-063 Table LIY. Summary of Moisture Determinations for Quail Albumen Samples Sponsor ID E01-0259-20506 E01-0259-20507 E01-0259-20520 E01-0259-20521 E01-0259-20509 E01-0259-20526 E01-0259-20514 E01-0259-20527 E01-0259-20515 E01-0259-20528 E01-0259-20529 E01-0259-20531 E01-0259-20532 E01-0259-20518 E01-0259-20533 E01-0259-20735 E01-0259-20736 EO1-0259-20737 E01-0259-20720 E01-0259-20738 E01-0259-20742 E01-0259-20743 EO1-0259-20744 E01-0259-20745 E01-0259-20746 EO1-0259-20747 E01-0259-20748 E01-0259-20749 E01-0259-20750 E01-0259-20751 Centre ID 0101988 0101992 0102044 0102048 0102000 0102068 0102020 0102072 0102024 0102076 0102080 0102088 0102092 0102036 0102096 0102904 0102908 0102912 0102844 0102916 0102932 0102936 0102940 0102944 0102948 0102952 0102956 0102960 0102964 0102968 Average % 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 C entre A nalytical L aboratories, Inc. Page 51 o f 102 000293 C entre Study No.: 023-063 Table LV. Summary of Moisture Determinations for Quail Yolk Samples Sponsor ID E01-Q259-20506 E01-0259-20507 E01-0259-20520 E01-0259-20521 E01-0259-20509 E01-0259-20526 E01-0259-20514 E01-0259-20527 E01-0259-20515 E01-0259-20528 E01-0259-20529 E01-0259-20531 E01-0259-20532 E01-0259-20518 E01-0259-20533 E01-0259-20735 E01-0259-20736 E01-0259-20737 E01-0259-20720 E01-0259-20738 E01-0259-20742 E01-0259-20743 E01-0259-20744 E01-0259-20745 E01-0259-20746 E01-0259-20747 E01-0259-20748 E01-0259-20749 E01-0259-20750 E01-0259-20751 Centre ID 0101985 0101999 0102041 0102045 0101997 0102065 0102017 0102069 0102021 0102073 0102077 0102085 0102089 0102033 0102093 0102901 0102905 0102909 0102841 0102913 0102929 0102933 0102937 0102941 0102945 0102949 0102953 0102957 0102961 0102965 Average % Moisture 48.0 43.0 46.0 45.0 45.0 49.0 49.0 47.0 54.0 47.0 47.0 41.0 49.0 50.0 47.0 47.0 46.0 47.0 41.0 43.0 44.0 43.0 45.0 53.0 46.0 47.0 46.0 45.0 45.0 43.0 C entre A nalytical Laboratories. Inc. Page 52 o f 102 000294 Centre Study No.: 023-063 FIGURES Centre Analytical Laboratories, Inc. Page 53 o f 102 000295 Centre Study No.: 023-063 F ig u re l . T v m ca i c a lib r a tio n c u r v e ror f f o s m\ \ \ \ \ \ m\ \ \ \ m 51 \ \ \ \ \ 1 12 i12 13 Lp r- r i 13 [o 14 l" S' rii i s Si L<m3! UI iI rf p r13 Lp Id m 13 ---,---- ----- ---- - 1 i 1................ ..... ............ ---- 1" ............... ----r--* 4 m i l i q # 8 ' j 6 < l '! 3 3 g 8 8 8 o m Tin* 1IV 000296 Centre Study N o.: 023-063 Figure 2. Chromatogram Representing a 0.1 ng/mL standard for PFOS AO*. Men AC. T im s .10 81/31/01 03 t* ltU 1 Nodlfladi w e h ia Factors oiaa n n ilw M i Ac m Tttroahalr RmT Mi LaLdde * rti S0o*#0,. Nftoditofchsti KM. N k R atio s Adj . R a tio s CM. v * i , M t l t t 000 a o lfttiv o RTs T13.U0* 39.30 a 10.0 4.43 0.30 0.01 3.00 4.00 1.00 X at. Typos t e t m l M Tlaos Ai m s OoioKti S ta rt Tlaas t a d TUMs 4.44 1470.3 3.MOO00: 4.14 4.30 C entre A nalytical Laboratories, Inc. Page 55 o f 102 000297 C entre Study N a : 023-063 Figure 3. Chromatogram Representing a 1.0 ng/mL standard for PFOS XMCSI SM U lYoci c w tm m u n Ac, t o u t M . tiaci 4 ttia n ri 1.060 o a m / 03.37.17 m im ittu t rw tn i N tlM n rw lM M i Ai m T h rc c to ld i i a . iM ctkit AT Kladov* f t i c c c c l R7i l* f . Vldtbt cp. lU libti tu p . fw tk ta tlo i tap . M L to tio i t* p . VAl. AMiCi BM M t U l v KTi Me 1 i.tc .3 3 30. 4.43 0.3 0.01 i.M 4.00 3.00 at* * i 1 1 la c . fypet M tcacitm t im i Arc** tt*lhl tu n ti i t a d TUMI f SMC 4.44 7347.4 n o w 1.73**001 4.3 at 4.41 a la 1TON tw o MOD 7300 taoo 1100 K mUll 0 m mU 000 400 300 300 too Centre Analytical Laboratories, Inc. Page 56 o f 102 000298 Centre Study N a : 023-063 Figure 4. Chromatogram Representing a Reagent Blank for PFOS (Centre ID Reagent Blank A, Set: 032601A) 03/31/01 Natl e u i aarBta F a et on Iteti T t e M b o I d i ktw TUMlWUi n r wtteiiowi BHa*iw.cacUadttaer.t Sate- Mietici tete> f M k M U o i bV. Mj . M(li late Vai. Baciai Q * a B a l e d * art AraciBaeoteciote f i a i t Molateti Barf Tiao* C entre A nalytical Laboratories, Inc. Page 57 o f 102 000299 Centre Study No.: 023-063 Figure 5. Chromatogram Representing Control Mallard Membrane for PFOS (Centre ID: 0003802 Blank A, Set: 032601C) C M n atn U aw M . Bate* AO. T lM i e .te a 03/17/01 0 ] i l l 107 M nf toapiuloopiiru to n UKMiMThAulMwAjmcldUl i AT MMOHI lH Q IICtCl *Tl M p. M ifiktt to o . Oook l a t U i to o . AdJ. JU tU i too Vi, Aaciot V M U e lv t KTi l1.H 11 9 .3 0 30.0 4.43 00..03 10 9.00 4.00 3.00' w e . Type JRtfMMIlM tlH TlM i KaiofcCi I t M C TUMI tod TlM i 4.44 1.44440*.0003 4 .3 7 4.11 t C entre A nalytical Laboratories, Inc. Page 58 o f 102 000300 Centre Study No.: 023-063 Figure 6. Chromatogram Representing Control Mallard Membrane Fortified with 10 ppb of PFOS, (Centre ID: 0003802 Spk A Dup. Inj., Set: 032601C) Centre Analytical Laboratories, Inc. Page 59 o f 102 000301 Centre Study N o.: 023-063 Figure 7. Chromatogram Representing Mallard Membrane Sample from Week 1 at 6.2 ppm ai (E01-0263-20825) (Centre ID: 0103092, Set: 032601C) Viaiypa> C eeeeecraeleei Naditladi laachlai raceart Mlucaia imf kcMn itloaaMldti mm. awet&e. RT Window Hafi,e t MideRaTi i U p- N tlfhc < tu p - M ek Ratio to . M J. R aclai ta p . V l. Raciei tfaa R o la tlv * RT< t a t . typa R acaatiea Tiaor Areas M iM i tot adr eTimTlas Centre Analytical Laboratories, be. Page 60 o f 102 000302 Centre Study No.: 023-063 APPENDIX A Study Protocol 00P-023-063 (Centre Study No. 023-063) and Amendments and Deviations C entre Analytical Laboratories, Inc. Page 61 o f 102 000303 000304 000305 000306 000307 C entreS tudy N a : 023-063 CejttreProtocolNa OOP-023-063 analysis of fortified samples to provide an indication of reproducibility. Fortification will be made to the matrix priorto extraction at levels tanging from theLOQto above the highestresidue found. Thefortification solution containing the test/reference substance will be applied to the designated samples via a Hamilton syringe, micropipette, pipet, or equivalent, to maintain consistency and accunuy. The average recovery andrelative standard deviation of the fortified samples will be 'rniteA Also, the average residue found and standard deviation for each matrixwill be calculated. The percent moisture will also be determined for each extracted sample if ampin remains afterextraction. Foreach matrix, 0.1 g samples will be weighed and dried in an oven at - 105C for -12-14 hours. Percent moisture for each ample will be determined in duplicate when there is sufficient sample available. H . PROTOCOL AMENDMENTS AND DEVIATIONS Any deviations from the protocol or from the analytical method as provided will be Hneifmenteri and reported promptly to the Study Director and Sponsor Representative. Planned changes to this protocol a to the analytical method will be mariain writing as an amendment and approvedby the Principal Investigator, the Study Director, and the Sponsor Representative. Any amendments or deviationswill be appendedto thisprotocol andincludedin thefinal report. 1 amendments: Planned changes to the approved protocol shall be ` ^^^^dbyM C Tdm ents that clearly describe the change,justification for the change,andimpacton the study. Amendments will be signed anddated by the Principal Investigator, StudyDirector, andSponsor^Representative. Copies of amendments will be sentto the qualitym a m rant. Z Protocol deviations: Protocol deviations, which are one time and unplanned deviations from the protocol, shall be documented in the studyrecords, noting the nature of the deviation, potential effect or impact on the study, and corrective action if required. Protocol deviations are signed anddated by the Principal Investigator, Study Director, and Sponsor Representative and reviewedby QAU. 12. RECORDS Records to be maintained include, but are not limited the following (as appropriate): 1. Sample tracking sheets) 2. Sample receipt records, storage history, andchains of custody 3. Historyandpreparationofstandards (stock, fortification, calibration) Centre AnalyticalLaboratories,foe. C entre A nalytical Laboratories, h e . Page 6 o f38 Page 67 of 102 000308 C entre Study No.: 023-063 CentreProtocol No. 00P-O23-063 4. Description of anymodifications to the method 5. Instrument run sheets,bench-sheets orlogs 6. Analytical datatables 7. Ail chromatographic and instrumental conditions 8. Sample extraction andanalysis dates ' 9. A complete listing of study personnel, signatures andinitials 10. Chronological presentation of ail studycorrespondence 11. Any datanecessary for the reconstruction of die study All chromatograms will contain the following: a. Sample identification, dat, arrow or other indication of the area of fotxyfr, andinjection numbercorrespondingto theran. b. Additionally, fortifications will include the fortification level of the analyte. c. Analytical standard chromatograms will additionally include the concentration (e.g* pg/ml, ng/mL, ppb, ppt, etc.). Each data set will contain information on temperatures, flow rates, column parameters, gases, instrument parameters, and instrument type, etc. The Centre study number will also be recorded on the first chromatogram of each daily analytical run. 13. QUALITY ASSURANCE Centre QA Unit will review the protocol, audit the study conduct and the study documentation (including raw data and final report) to ascertain that all GLP standards are met Centre QA Unit will inspect the study at intervals adequate to assure compliance to GLP's, and will report the Endings of the audits to the Principal Investigator. CentreManagement, andthe StudyDirector. 14. DATA AND REPORT 1. All raw data and the original signed protocol will be maintained in the study file. This data includes the protocol amendments, protocol deviations, laboratory notebooks, analytical standard solution preparation, sample chain of custody sheets, sample work sheets, chromatograms, calibration curves, a n d a n y other appropriate data generated. Raw datanot used will be stored in thestudy file. The reason forexclusion will be documental 2. A report will be issued by Centre, approved and signed by the Study Director and the Sponsor Representative and sufficient for submission to EPA. The reportcontents shouldinclude, but not limited to: 1. Objectives andprocedures statedin the protocol 2. Analytical andstatistical methods used Centre A nalytical Laboratories, lac. C entre A nalytical Laboratories, Inc. Page7 of38 Page 68 o f 102 000309 Centre Study N o.: 023-063 CentreProtocolNo. OOP-023-063 3. materialsidentifiedbyname, lot,purity, andother characteristics 4. Nameofperforminglaboratoryandanalytical startandterminationdates 3. Statementpreparedand signedby the qualityassuranceunit 6. Tables containingall applicabledata 7. All chromatographicand instrumental conditions 8. A completelistingof Centrestudypersonnel Centre will send a copy of the draft report to the Study Director and Sponsor Representative, who will return the reportwith comments. Centre Analytical will make revisions and finalize the report with the approval of the Study Director and SponsorRepresentative. The Centre QA unit will conduct an audit of tbe final draft report and data files to assure accuracy and GLP compliance. The report must be approved by the Study Director and Sponsor Representative prior to signature. A statement of QAU inspections and a statement of GLP compliance will beincludedin tirefinalreport Any corrections or additions to the final report shall be in the form of an amendment by tire Principal Investigator. The amendment shall clearly identify the part of the final report that is being corrected and die justification for correction. The amendment shall be signed and dated by the Principal Investigator, StudyDirectorandSponsorRepresentative. REPORTDISTRIBUTION: Original: StudyDirector Copy. CentreArchives IS . ARCHIVE STATEMENT Study records to be maintained: Records to be maintained for die study jjj raw data, observations recorded during tbe conduct of the study, documentation chromatograms, specimen tracking information, and study related correspondence.' This includes a description of equipment used during the conduct of the study. All "*--*!hi mAanvshinning records shallbe retained. Document archives: Upon completion of the study, the study records, protocol and amendments, and the final report and amendments shall be retained in the 3M document archives. If it is necessary to substitute acopy foran original record, it will be certified as an exact copy. Centre wilt retainfacility-related originals and a copy of the final reportandrawdatapackage. 16. REFERENCES 1 U.S. Environmental Protection Agency. Toxic Substances Control Act (TSCA). GoodLaboratoryPractice Standards. FinalRule,40 CFRPart792. Centre Analytical Laboratories, Inc. C entre A nalytical Laboratories, Inc. Page 8 of38 Page 69 o f 102 000310 Centre Study No.: 023-063 CentreProtocol No. OOP-023-063 17. PROTOCOLAPPROVAL This protocol was audited by the Quality Assurance Unit of Centre Analytical Laboratories, be. QualityAssuranceReviewer, Centre _______ tU fJL-- U t/" Naomi Lovallo, PhD. Quality AssuranceAuditor Principal Investigator, Centre 3 /Wt>/ Kte FacilityManagement, Centre .Jbhn Flaherty Operations Manager Study Director, WHdIlfe International, Ltd. ALf- J U ^ l . Senior Biotogst Sponsor, 3M -sA i f >te ' Centre Analytical Laboratories, Inc. C entre A nalytical Laboratories, lac. Page 9 of 38 Page 70 o f 102 000311 Centre Study No.: 023-063 Cwne Protocol No. OOP-023-063 APPENDIX: ANALYTICAL METHOD A. "Determination ofPerSoorooctaaesulfonatein EggMembrane, Albumen, and Yolkby LC/MS1MS", Centremethod number0OM-O23-O15 Centre Analyticallaboratories, lo t C entre A nalytical Laboratories, Inc. Page 10 of 33 Page 71 o f 102 000312 C entre Study No~ 023<463 CentreProtowlNo. 00^023^)63 I TITLE AUTHORS Emily S o n fe m iW mHiheny : . - : DAtElSSOEp . Jm&ay2,2001 SPKBSE 3MBiTBOBBicnnirihontcij , Bufldhij2-3E-09 FO B 33331 &.J>i4MHiS133i3331 P er fo r m in g l a bo r a t o r y CentreAnalyticalLaberatorio,he. (Centre) 3043 Research E rin Sate'CoBqe,PA 16101 5EHEE n'ia'Bofii:. 0034U CENTRE METHODNUMBER 0MB-01S t^ ' TOTAL NUMBER OF PAGES 23 : CentreAnalyticalOBrtbri"'Lnicr C entre A nalytical Laboratories, Inc. Page H of38 Page 72 o f 102 000313 Centre Study No.: 023-063 .C om e Protocol N0.OOP-O23.063. * ' .. MANAGEMENT APPROVAL. * * . pg^iC FR 7913. method dtwlcjaacatU n rrqntol tobe cnafarr^i^. _ .. '^C V xxl LaboratoryP n c . Bowevc; the wotfcwet in confer----- L?!!" 1* 1 w od d op etai| procdera*andjeaeralG lPie|nW qiH . W icable ^ u & / '^tbl Stauffer Pnoc^lbrnaptor. ' .Date CcotraAnaljtieal L3boi3torio .h e . ^ - ft- r fth T M / ' Date laboratoryMaaaj^ Croire AiidydcdLiboraioris,iBC> . -------- y g * -- ^ . R ieh roij^ fdn im .P h i). . PieideaL- Centre Analytical Labontoria, h e . f` Dare , . f Rcpnsrnativg . ' SMEeemanental Laboratory ' CanAati]^Lte3aries.lae.SadjrtJtD34U Centre Analytical Laboratories; lu e' C entre A nalytical Laboratories, Inc. a*e2 ' Page 12 of 38 Page 73 o f 102 000314 Centre Study No.: 023-063 -CentrePraoedNa 00P-23-O63 TABLEOFCONTENTS TnUl4GE_ "H ^G EM EN T APPROVAL. TABLE OF CONTENTS. uetw tabus. LISTOVIGDHES. L SOMMAS?__ 1 EZPESJMENTAL'COMPpwbs. L CHEMICALSANDSU PPLES-i- U-Omdu. U**&JIWWWMUITBWt, .... BJmmirMmSMiMs AA Sam oa__:-----------------:-------------- :------ - JAU- jUnUn..aMxiBr&Ha.MhvSfc_io_a_i.f_tt-*--c-k-V--B-i-L-M--O- -M--i-K-m---sd:_im__s_ie___1 All frrffcfcnSulnimf. ; All r.nimlmT^fctrft < ETH0D FuMTQwaaM. f'A. AuauFnouno. luauEeacnw fcMr - Ami-nairlCHOalS. .. - ------. .....' O pintCmM m i(TBpalrm^eijl. << Eatflt1kmnftnmata------ :-------- :-- ; lr n fc h fc - T--yli tufrjii *--At*eaia'iuiinNwcxuQgBMBBWn-.1. A. Tyuam*BKmMiLm- * CALCULATIONS;---- SAETT. TABUS___ FIGURES__ CmAfc]rialubnja.'tK.Sm ififiO W -7 --7 -7 - 1 3 Centre AnalyticalIaboiticnTIhi' C entre A nalytical Laboratories. Inc. Paje 13 of38 Page 74 o f 102 000315 ................ C eutreS tudy No.: 023-063 : OsanePtotocola 00P-O23-OS3 LISTOFXAJBLES ' 000316 000317 000318 000319 000320 000321 000322 000324 000325 000326 000327 000328 000329 000331 000332 000333 C00334 000335 C entre Study N o.: 023-063 C^ftW teoINo. OOP-023-063 CourtMethodNo:OOM-^CJ^XS' Psurt3: Bmoefcdfte Chromat}niniofa QuailE jYolkControl --- --CentreAnalytkilibapioriei.Inc-- -,-------C entre A nalytical Labratorie, Inc. . Page 34 of38 Page 95 o f 102 000336 C entre Study N o.: 023-063 ' CHtaRBtSMiNo:OOP-023-063 CentreMethodNo:OttM.023415 Figure5: R epraentalire Chromatogram ofa Qoafl Eg* Albumen Coittrol ;----DrcTOAnafyBcatLaboratories,Idc.- - C entre A nalytical Laboratories, In c ., : feras Page35 of 38 Page 96 o f 102 000337 l > Centre Study No.: 0234)63 "^CrPiiocoWfc 0QP-023-O63 Ftp 7: IUprts*ntafinCfcronmn>jramofaQnaBE--Menbran*Control * FortffledatI#n'j(ppW. .I mm * Jj 5 tata tofcfactnci i(Q ata "".lui:1.l-i . . U ir FortffledatU ngfetppb) CwA*^Ulwiate.&*r*fl&OtJ Centi? Ai^yticalLboratnes, inc. CentreAnalytical Laboratories, Inc. F>is26 , Page36 of38 Page 97 o f 102 000338 C entre Study N o.: 023-063 "Cato'Procr3%rDpPJ323^)63 CotttMtthodNo:0aM-023mij ! R p m y . tp m ta ta re Q roniatograni ofa QuaB Eg*Tft Contra! Ffcttfledat 1&0nf^tippt>) -- CentreAnsJytkalLabotatoria,Inc. Centre Analytical Laboratories, Inc: Page37of38 P age98ofl02 000339 C entre Study N o.: 023-063 : CentProtocolNo. 00P-O23-063 . ' CnUreMethodKe; OOM-O2ioi5, ***'* tl! K ipratalalhre QtromatogniO ofa Qa a BAHr a j a e , Contni . FortU M atlO JosCppb) (^ fM tJial3haan.MSalfimJiiS Centre AnalyticalLaboratories, Inc. C entre A nalytical Laboratories, Inc, Page38 of 38 Page 99 o f 102 000340 C entre Study No.: 023-063 MHS Besaurh Drire, State Cot*, PA IM01. Phone (SUH31-M32, Foolm&e (*1<)23M2J3 PROTOCOL AMENDMENT Amendment Number 1 Effective Date: 3/23/01 Page 1 of 1 Centre StudyNumber 023-063 Centre Protocol Number 00P-023-063 1. Title DESCRIPTIONOF AMENDED SECTION 000341 Centre Study N a : 023.-063 3048Km irdi DHm StmCttUew. PA IMO!. J?boggJSKtt31-M 32. F ao tafle 1*141231.12 PROTOCOL AMENDMENT Amendment Number 2 Effective Date: 3/2&01 Page 1 of 1 Centre Study Number 023-063 Centre Protocol Number 0OP-O23-O63 DESCRIPTION.QEAj lactam ECTIQN 1. Section 9: AnalyticalMethod 2. Section 9: Analytical Method 3. Section 10: Experimental Design AMENDEP.TO 1. Amend Page 21 Section 4.3.2 a, to read: Priorto weighing andextracting the egg membrane samples, the mranKran; will be rinsed with ASTMType IH2O and patted dry. 2. OnPage 24 Section 4.4.4.d, remove: One ASTM TypeT water blank 3. Amend Page 6 lastparagraph, second sentence to read: Foregg yolk andalbumen,- 0.5 g samples and eggmembrane - 0.1 g samples will be weighed anddriedin anoven at - 105Cfor.- i2-l'4 hours. RATIONA! B 1. At the instruction of the study directorto ensure that no contamination of egg yolk or albumen is present in the sample. Z This sample is only to check for PFOS residue present in the ASTM Type IH20 used. 3. To account for sample availability. IMPACTONTHE STUDY 1-3. No negative impact on the study. Study DMirreeccttoorr Silganature ai Date Dai CentreQAUReview &L (I Febnwry 12,1998/1 Centre A nalytical Laboratories, Inc. Page 101 o f 102 000342 Centre Study No.: 023-063 3MSB*aeardiDrivenStata CaOefe, FAMMI. tourna (4) 231-M32,Ftalm ik: (SW)23I-US3 PROTOCOL DEVIATION DeviationNumber: i DateofOccurrence: (1) 03/27AH. (2)04/02/01 Centre StudyNumber: 023-063 CentreProtocol Number OOP-023-063 BESCRIFnON OFDEVIATION 1. 8 Analytical Method4.4.4.f. QuantifiedCentresample0103196andits duplicateinjection when the concentrations weregreaterthanthatof thehigheststandardinSet032601C 2. SAmendment!: Acceptedrecoveries of58%and61%forCentresample0003800SpkAandits duplicate injection in Set 033001G ACTIONS TAKEN 1 neadment itmed. SOP mriiioa. tic - 1-2. Protocoldeviationissued. RecordedBy/Date: A M s . ^ i. v i wIMt nPA< C- rTnOu NT Tu Hc Ee SnTmUDv Y 1. No negativeimpactbecausetheareaswere<5%greaterthanthehigheststandard. 2. NonegativeimpactbecauseSpkBgaveacceptablerecoveriesandsampleresults reportedinthe set were only forconfirmation. DetJiLes Principal InvestigatorSignature Study Director Signature ....... " Date 1 J G|>V[o| Date SpoggpafrfinagfaaeatSignKue s' Date 1 Mo' CALQAUReview fi k Mil February 1Z 199S/2 Centre A nalytical Laboratories, Inc. Page 102 o f 102