Document NEk9No8dyj9nYrGQKRO0RD5RV

BACK TO MAIN SUMMARY OF ANALYTICAL RESULTS: MULTI-CITY FOOD STUDY The Multi-City Study was originally designed by Battelle Memorial Institute (Columbus, OH), to obtain preliminary data about the presence of fluorochemicals in foods and in drinking water to understand the potential sources of human exposure. The Multi-City Study paired each of three cities having manufacturing or commercial use of fluorochemical products (test cities) with three cities that do not (control cities). Information on residues in selected foods was obtained from analyses of food samples collected in a market basket study. The market basket sampling of the original MultiCity Study design was implemented by Pace Analytical Services, Inc., Minneapolis, MN. The samples were analyzed for PFOS, PFOA, and FOSA by Centre Analytical Laboratories, Inc., State College, PA. The distributions of the PFOS, PFOA, and FOSA residue data by food and city category reveal similar patterns of residue concentrations in the control and test cities for each type of food. A total of 12 samples were found to contain levels of fluorochemical residues above the limit of quantification. Of the 12 samples with measurable fluorochemical residue levels, eight were samples collected in test cities. Measurable quantities of PFOS were found in five samples: four whole milk samples (three from test cities) and a ground beef sample (test city). PFOS residues found in the foods ranged from non-quantifiable levels to 0.852 ng/g. Measurable quantities of PFOA were found in seven samples: two ground beef samples (neither from test cities); two bread samples (one from a test city); two apple samples (both from test cities); and one green bean sample (from a test city). PFOA residue levels ranged from non-quantifiable levels to 2.35 ng/g. A value of 14.7 ng/g was found for PFOA in a bread sample from a control city, but was considered "suspect" by Centre Analytical. BACK TO MAIN ANALYTICAL REPORT STUDY TITLE Analysis of PFOS, FOSA and PFOA From Various Food Matrices Using HPLC ElectrosprayMass Spectrometry DATA REOUrREMENTS EPA TSCA Good Laboratory Practice Standards 40 CFR 792 STUDY DIKECTOR Karen Smith, Centre SPONSOR REPRESENTATIVE Susan A. Beach, 3M ANALYTICAL REPORT COMPLETION DATE June 21,2001 PERFORMING LABORATORY / TESTING FACILITY Centre Analytical Laboratories, Inc. (Centre) 3048 Research Drive State College, PA 16801 Phone: 8 14-23 1-8032 STUDY SPONSOR 31 Environmental Technology and S a d y Services Building 2-3E-09 PO Box 33331 St. Paul, MN 55133-3331 PROJECT IDENTIFICATION Centre Protocol Number: OOP-023-057 Centre Study Number: 023-057 Total Pages: 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT Centre Study Number 023-057, entitled "Analysis of PFOS, FOSA and PFOA From Various Food Matrices Using HPLC ElectrosprayNass Spectrometry,''conducted for 3M Environmental Technology and Safety Services, was performed in compliance with EPA TSCA Good Laboratory Practice Standards (40 CFR, Part 792) by Centre Analytical Laboratories, Inc. with the following exceptions: 1. The FOSA reference substance (analytical standard) used in this study has not been characterized under GLP's. 792.105(a). 2. Specimen (egg sample) identification for three samples was not according to 792.130(c) because the labels became removed from the containers. I KarenSmith " Study Director Centre Analytical Laboratories, Inc. CblZllO I Date Susan A. Beach Sponsor Representative 3M Environmental Technology and Safety Serv:ices Centre Analytical Laboratories, Inc. Page 2 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 QUALITY ASSURANCE STATEMENT Centre Analytical Laboratories' Quality Assurance Unit reviewed Centre Study Number 023-057, entitled, "Analysis of PFOS, FOSA and PFOA From Various Food Matrices Using HPLC Electrosprayhlass Spectrometry". All phases were reviewed for conduct according to Centre Analytical Laboratories' Standard Operating Procedures, the Study Protocol, and all applicable Good Laboratoiy Practice Standards. All findings were reported to the Study Director and to management. Phase 1. Protocol Review Date Inspected 02/14/01 2. Extraction and Fortification 02/14/01 Date Reported to Studv Director* 03/23/01 03/23/01 Date Reported to Centre Management 02/26/0 1 02/26/0 1 Date Reported to Sponsor Management 03/23/0 1 03/23/01 3. Raw Data and Draft Report Review 04110-13101 0512 1/01 06/04/0 1 06/15/01 4. Raw Data and Draft Report Review 05/30/01 0610 1 /O 1 06/04/0 1 0611510 1 5. Final Report Review 06/19/01 06/20/0 1 06/21/01 0612 1 /O 1 *The Study Director resided at the sponsor until 05/01/01 at which time a protocol amendment transferred Study Director duties to Centre Analytical Laboratories. u-1, *p e--- Naomi Lovallo Senior Quality Assurance Auditor - 6 /a-ih Date Centre Analytical Laboratories, Inc. Page 3 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 CERTIFICATION OF AUTHENTICITY This report, for Centre Study Number 023-05'7, is a true and complete representation of the raw data for the study. Submitted by: Centre Analytical Laboratories, Inc. 3048 Research Drive State College, PA 16801 (814) 231-8032 IStudy Director, Centre: A' Study Director Centre Analytical Laboratories, Inc. J&I21Jo I Date Centre Analytical Laboratories, Inc. Facility Management: $hn Flaherty / - Date Laboratory Manager Centre Analytical Laboratories, Inc. Spoyor Representative, 3M Suss: A. Beach I - Date 3M Environmental Technology and Safety Services Centre Analytical Laboratories, Inc. Page 4 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 STUDY IDENTIFICATION Analysis of PFOS, FOSA and PFOA From Various Food Matrices Using HPLC ElectrosprayMass Spectrometry CENTRE PROTOCOL NUMBER: OOP-023-057 TYPE OF STUDY: Residue TEST SYSTEM: Green beans, apples, pork muscle, cow milk, chicken muscle, chicken eggs, bread, hot dogs, catfish and ground beef TEST MATERIAL: PFOS, FCSA and PFOA SPONSOR: 3M Environmental Technology and Safety Services Building 2-3E-09 PO Box 33331 St. Paul, MN 55133-3331 STUDY DIRECTOR: Karen Smith Centre Analytical Laboratories, Inc. Phone: (814) 231-8032 TESTING FACILITY: Centre Analytical Laboratories, Inc. 3048 Research Drive State College, PA 16801 ANALYTICAL PHASE TIMETABLE: Study Initiation Date: Analytical Start Date: Analytical Termination Date: 0 1/09/0 1 01/11/01 05/22/01 Centre AnalyticalLaboratories,Inc. Page 5 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: 0013-023-057 PROJECT PERSONNEL The Study Director for this project at Centre Analytical Laboratories, Inc. was Karen Smith. The following personnel from Centre Analytical Laboratories, Inc., were associated with various phases of the study: Name Karen Smith Emily Stauffer Angela Morgan Tiffany Proctor Rickey Keller Lawrence Ord Dave Bell Sharareh Zolghadr Jason Farabaugh Mitra Arjmand Yu Qiong Tan Ling Ling Liu Xlaoming Zhu Title Scientist Scientist Technician Technician Sample Custodian Sample Custodian Scientist Technician Scientist Technician Technician Technician Technician Centre Analytical Laboratories,Inc. Page 6 of 151 . BACK TO MAIN . Centre Study No. 023-057 Centre Protocol No: OOP-023-057 TABLE OF CONTENTS TITLE PAGE ...................................................................................................................... 1 GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT ............................. 2 QUALITY ASSURANCE STATEMENT.......................................................................... 3 CERTIFICATION OF AUTHENTIClTY........................................................................... 4 STUDY IDENTIFICATION............................................................................................... 5 PROJECT PERSONNEL.................................................................................................... 6 TABLE OF CONTENTS .................................................................................................... 7 LIST OF TABLES .............................................................................................................. 8 LIST OF FIGURES ............................................................................................................. 9 LIST OF APPENDICES ................................................................................................... 11 1.0 SUMMARY ............................................................................................................... 12 2.0 OBJECTIVE................................................................................................................ 12 3.0 INTRODUCTION....................................................................................................... 12 4.0 TEST SYSTEM ......................................................................................................... 13 5.0 REFERENCE MATERIAL ....................................................................................... 14 6.0 EXPERIMENTAL DESIGN ...................................................................................... 15 7.0 DESCRIPTION OF ANALYTICAL, METHOD ....................................................... 15 7.1 Extraction Procedure ........................................................................................... 15 7.2 Preparation of Standards and Fortification Solutions .......................................... 15 7.3 Chromatography .................................................................................................. 16 7.4 Instrument Sensitivity .......................................................................................... 17 7.5 Description of Instrument and Operathg Conditions.......................................... 17 7.6 Quantitation and Example Calculation ................................................................ 18 8.0 RESULTS AND DISCUSSION............................................................................... 20 9.0 CIRCUMSTANCES THAT MAY H A W. AFFECTED THE DATA .................... 21 10.0 RETENTION OF DATA AND SAMPLES............................................................. 21 11.0 TABLES ................................................................................................................... 22 12.0 FIGURES ................................................................................................................. 41 13.0 APPENDICES.......................................................................................................... 65 Centre AnalyticalLaboratories. Inc. Page 7 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 LIST OF TABLES Table I. Summary of PFOS, FOSA and PFOA Fortification Recoveries ........................ 23 Table II. Summary of PFOS, FOSA and PFOA Residues Found in Samples .................26 Centre Analytical Laboratories, Inc. Page 8 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: 00P-023-057 LIST OF FIGURES Figure 1. Typical Calibration Curve for PFOS.............................................................. 42 Figure 2. Typical Calibration Curve for FOSA ............................................................. 43 Figure 3. Typical Calibration Curve for PFOA ............................................................. 44 Figure 4. Chromatogram Representing a 0.5 ng/mL mixed calibration standard of PFOS, FOSA and PFOA ................................................................................ 45 Figure 5. Chromatogram Representing a 2.5 ng/mL mixed calibration standard of PFOS, FOSA and PFOA ................................................................................ 46 Figure 6 . Chromatogram Representing a Control Apple Sample (Centre ID: 0009347 Blank Al, Data Set: 020901C Apple)............................................................ 47 Figure 7. Chromatogram Representing a Control Apple Sample Fortified with 2.5 ng/g of PFOS, FOSA and PFOA (Centre ID: 0009347 Spk A l , Data Set: 020901C Apple) ............................................................................. 48 Figure 8. Chromatogram Representing an Apple Sample (Centre ID: 0012927, Sponsor ID: Publix, 18257, Data Set: 020901C Apple) ................................ 49 Figure 9. Chromatogram Representing a Control Green Bean Sample (Centre ID: 0009346 Blank A2, Data Set: 020601C Bean) .............................................. 50 Figure 10. Chromatogram Representing a Control Green Bean Sample Fortified with 2.5 ng/g of PFOS, FOSA and PFOA (Centre ID: 0009346 Spk A2, Data Set: 020601C Bean)............................................................................... 51 Figure 11. Chromatogram Representing a Green Bean Sample (Centre ID: 0012910, Sponsor ID: Bruno's, 18341,Data Set: 020601C Bean)................................ 52 Figure 12. Chromatogram Representing a Control Pork Muscle Sample (Centre ID: 0009348 Blank Al, Data Set: 011501A Pork)............................................... 53 Figure 13. Chromatogram Representing a Control Pork Muscle Sample Fortified with 2.5 ng/g of PFOS, FOSA and PFOA (Centre ID: 0009348 Spk A l , Data Set: 011501A Pork) ............................................................................... 54 Figure 14. Chromatogram Representing a Pork Muscle Sample (Centre ID: 0012795, Sponsor ID: Winn-Dixie, 18227,Data Set: 011501A Pork).......................... 55 Figure 15. Chromatogram Representing a Control Milk Sample (Centre ID: 0009349 Blank Al, Data Set: 020701A Milk) ............................................................. 56 Figure 16. Chromatogram Representing a Control Milk Sample Fortified with 2.5 ng/g of PFOS, FOSA and PFOA (Centre ID: 0009349 Spk A l , Data Set: 020701A ~ l k...)............................................................................ 57 Centre Analytical Laboratories, Inc. Page 9 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 LIST OF FIGURES (CONTINUED) Figure 17. Chromatogram Representing a Milk Sample (Centre ID: 0012871, Sponsor ID: Winn-Dixie, 18202, Data Set: 020701A Milk) ......................... 58 Figure 18. Chromatogram Representing a Control Chicken Egg Sample (Centre ID: 0009342 Blank A3, Data Sets: 050901A Egg for PFOS, 012401A Egg for FOSA, and 052101A Egg for PFOA)....................................................... 59 Figure 19. Chromatogram Representing a Control Chicken Egg Sample Fortified with 2.5 ng/g of PFOS, FOSA and PF0,4 (Centre ID: 0009342 Spk A3, Data Sets: 050901A Egg for PFOS, 012401A Egg for FOSA, and 052101A Egg for PFOA) ............................................................................... 60 Figure 20. Chromatogram Representing a Chicken Egg Sample (Centre ID: 0012861, Sponsor ID: Save, 18167 and 21224, Data Sets: 050901A Egg for PFOS, 012401A Egg for FOSA, and 052101A Egg for PFOA) ............................... 61 Figure 21. Chromatogram Representing a Control Bread Sample (Centre ID: 0009343 Blank Al, Data Set: 020901A Bread) ............................................................ 62 Figure 22. Chromatogram Representing a Control Bread Sample Fortified with 2.5 ng/g of PFOS, FOSA and PFOA (Centre ID: 0009343 Spk A l , Data Set: 020901A Bread) ............................................................................. 63 Figure 23. Chromatogram Representing a Bread Sample (Centre ID: 0012887, SDonsor ID: Albertson's. 18244. Data Set: 020901A Bread) ........................ 64 Centre Analytical Laboratories, Inc. Page 10 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 LIST OF APPENDICES & Appendix A Study Protocol OOP-023-057 (Centre Study No. 023-057), Amendments and Deviation ...................................................................... 65 Centre Analytical Laboratories, Inc. Page 11 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 1.0 SUMMARY The purpose of this study was to analyze residues of PFOS, FOSA and PFOA in various food matrices as specified in Centre Protocol OOP-023-057. The analytical method used for this study was entitled, "Method of Analysis for the Determination of Fluorochemicals in Green Beans, Apples, Pork Muscle, Cow Milk, Chicken Muscle, Chicken Eggs, Bread, Hot Dogs, and Catfish by LCMSMS." The limit of quantification for PFOS, FOSA and PFOA in food was 0.5 ppb (ng/g). Average fortification recoveries and relative standard deviations were 101% f 9.0% for PFOS, 96% k 13% for FOSA and 104% f 14% for PFOA. Residues ranging from non-quantifiable levels to 0.852 ppb (ng/g) for PFOS, nonquantifiable levels for FOSA, and non-quantifiable to 2.35 ng/g for PFOA (one value of 14.7 ppb (ng/g) was found for PFOA but was Considered suspect) were found in the food samples. 2.0 OBJECTIVE The objective of this study was to determine levels of PFOS, FOSA and PFOA in food samples collected from local grocery stores from six test cities outlined in 3M Quality Assurance Project Plan for Empirical Human Exposure Assessment Multi-City Study Sampling Task using the analytical method entitled "Method of Analysis for the Determination of Fluorochemicals in Green Beans, Apples, Pork Muscle, Cow Milk, Chicken Muscle, Chicken Eggs, Bread, Hot Dogs, and Catfish by LCMSMS." 3.0 INTRODUCTION This report details the results of the residues of PFOS, FOSA and PFOA detected in green beans, apples, pork muscle, cow milk, chicken muscle, chicken eggs, bread, hot dogs, catfish and ground beef, using the analytical method entitled, "Method of Analysis for the Determination of Fluorochemicals in Green Beans, Apples, Pork Muscle, Cow Milk, Chicken Muscle, Chicken Eggs, Bread, Hot Dogs, and Catfish by LCMSMS." Complete details of the analytical methodology can be found in Appendix A. The study was initiated on January 09, 2001, when the study director signed the protocol OOP-023-057. The complete protocol and amendments can be found in Appendix A. The analytical start date was January 11, 2001, and the analytical termination date was May 22,2001. Centre Analytical Laboratories, Inc. Page 12 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 4.0 TEST SYSTEM The food samples analyzed in this study were received frozen from Pace Analytical on December 09, 2000 and stored frozen (c-10C) upon receipt. Pace Analytical collected the samples for 3M Environmental Laboratory. The samples were then logged in on December 18, 2000 by Centre personnel and transferred to different frozen storage locale. All of the samples were processed according to the method, "Method of Analysis for the Determination of Fluorochemicals in Green Beans, Apples, Pork Muscle, Cow Milk, Chicken Muscle, Chicken Eggs, Bread, Hot Dogs, and Catfish by LCMSRMS." Exact details of processing can be found on page 91 of this report. After processing, the samples were stored in a freezer at a temperature of -10C or below until used and placed back into a freezer immediately after use. At the request of the sponsor, the egg samples were returned frozen to 3M Environmental Laboratory on February 26, 2001 for possible reanalysis. The egg samples were returned unanalyzed from 3M Environmental Laboratory to Centre Analytical Laboratories, Inc. These samples were received frozen from 3M Environmental Laboratory on May 03, 2001 for PFOS and PFOA re-extraction and analysis. They were assigned different 3M and Centre ID'S. The food samples used for the controls and matrix spikes were the same samples used in the method validation study (Centre Study No. 023-043). They were purchased from Giant, Store #111 in State College, PA and assigned the following sample IDS upon log in at Centre: I CentreSarnDle~~I 0009341 Samde DescriDtion I Chicken Muscle Chicken Eggs ~ 0009344 0009345 Hot Dogs Catfish Green Beans Pork Muscle Whole Milk Ground Beef All of the control samples were purchased on September 27, 2000 except for the ground beef, which was purchased on January 03, 2001. The samples were stored in a refrigerator (4" k 2OC) until processed. All of tlhese control samples were processed with dry ice in a Hobart food chopper on October OZ!, 2000 except for the ground beef, which was processed on January 03, 2001. After processing, the samples were stored in a freezer at a temperature of -10C or below until used and placed back into a freezer immediately after use. Centre Analytical Laboratories, Inc. Page 13 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Sample login 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 PFOS was received at Centre on June 05, 2000, FOSA was received on February 15, 2000 and PFOA was received on July 06, 2000 from 3M Environmental Technology and Services. Characterization of the reference materials PFOS and PFOA was performed at Centre on August 31,2000 and November 06,2000, respectively, and documentation can be found in raw data package associated with this report. The characterization of FOSA is in progress and the necessary information will be added upon complete characterization of the reference materials. All analyses were completed prior to standard expiration dates. The available information for the reference material is listed below. FOSA was stored at room temperature and PFOS and PFOA were stored frozen at I -1OOC. Compound PFOS FOSA PFOA TCR Substance No. TCROOO 17-46 SD-029 TCR99030-30 Batch No. NA L- 15709 332 Purity (%) 97.9 95.1 95.0 Expiration Date 0813 1/01 02/15/01 10/31/01 Molecular structures of PFOS, FOSA and PFOA are given below. PFOS Chemical Name - Molecular weight = 0 Perfluorooctane sulfonate 499 (CsF17S03-) Note: The neutral molecule and standard form which PFOS (anion) is derived from is potassium perfluorooctane sulfoinate [ C S F ~ ~ S O ~mKo]le, cular weight 538. FOSA Chemical Name - - Molecular weight = 0 Perfluorooctane sulfonylamide 499 Centre Analytical Laboratories, Inc. Page 14 of 151 PFOA Chemical Name - - Molecular weight = BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Perfluorooctanoate 413 Note: The neutral molecule and standard form which PFOA (anion) is derived from is ammonium perfluoroctanoate [C7F&OONH4], molecular weight 43 1. 6.0 EXPERIMENTAL DESIGN Each sample was extracted in duplicate. A set contained one matrix blank and two matrix blanks spiked with known amounts of PFOS, FOSA and PFOA and 6 - 10 samples. The re-extracted egg sets also included one reagent blank. The extracts were analyzed by LCMSMS. Samples with residues below the linear range of the calibration curve were not quantified; however, residues in the control samples were calculated up to 20% of the LOQ by extrapolation of the calibration curve, in order to correct fortification recoveries. 7.0 DESCRIPTION OF ANALYTICAL METHOD Analytical method entitled "Method of Analysis for the Determination of Fluorochemicals in Green Beans, Apples, Pork Muscle, Cow Milk, Chicken Muscle, Chicken Eggs, Bread, Hot Dogs, and Catfish by LCMS/MS" was used for this study. 7.1 Extraction Procedure Five grams of processed sample were weighed into 50 mL polypropylene centrifuge tubes and fortified (if necessary). The samples were extracted with ACN followed by a SPE column (florisil, silica gel, carbon and LC-NH2) clean up. The column eluate was collected and concentrated to near dryness. Final volume was adjusted to 2 mL with 2% ascorbic acid in methanol. The samples were analyzed using electrospray LCMSMS. 7.2 Preparation of Standards and Fortification Solutions Standard stock solutions of PFOS, PFOA and FOSA were prepared on September 18, 2000, September 26, 2000 and October 03, 2000, respectively. Fortification and calibration standard solutions were prepared on October 17, 2000 as specified in Centre Analytical Laboratories' analytical method entitled, "Method of Analysis for the Determination of Fluorochemicals in Green FJeans, Apples, Pork Muscle, Cow Milk, Centre Analytical Laboratories, Inc. Page 15 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Chicken Muscle, Chicken Eggs, Bread, Hotdogs, and Catfish by LC/MS/MS." Individual stock standard solutions of PFOS, FOSA and PFOA were prepared at concentrations of 100 pg/mL by taking 10 mg of the standard (corrected for purity and salt content where applicable) and bringing it up to 100mL with methanol. From these solutions, a 1pg/mL mixed fortification standard solution was prepared by taking 1 mL of each stock and bringing the volume up to 100 mL with methanol. The 1 pg/mL mixed fortification standard was diluted ten-fold with methanol to make the 0.1 p g / d mixed fortification standard. Then, the 0.1 pg/mL mixed standard was diluted ten-fold with methanol to make the 0.01 pg/mL fortification standard. LC/MS/MS calibration standards were prepared in methanol at the following concentrations 50,25, 10,5.0,2.5, 1.0, and 0.5 ng/mL according to the following table: Initial Concentration Volume Diluted to Final concentration 1000 5.0 1010 50 1000 2.5 100 25 10.0 1 10.0 100 1.o 5 .O 10.0 100 0.5 New standard stock, fortification and calibration solutions were made for PFOS and PFOA in order to perform the egg re-extraction and analysis. The PFOS standard stock solution was prepared on March 14, 2001 and the PFOA standard stock solution was prepared on April 11, 2001. Mixed fortification and calibration solutions of PFOS and PFOA were prepared from the stock solutions `onMay 03,2001 as described above. The only modification from the above procedure is that FOSA was omitted from the mix since the re-extraction and analysis was for PFOS and PFOA only. The stock standard solution and all fortificaticln and calibration standard solutions were stored in a refrigerator (4" k 2C) when not in use. Documentation of standard preparation can be found in the raw data associated with this report. 7.3 Chromatography Quantification of PFOS, FOSA and PFOA was accomplished by LC/MS/MS analysis using electrospray LC/MS/MS. The retention times of PFOS, FOSA and PFOA were - 5.4 min, - 6.0 min and - 5.3 min., respectively. The matrices Qd not contribute any interfering peaks corresponding to the Centre Analytical Laboratories, Inc. Page 16 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 analyte retention times; however, there were some occasions where the control matrices appeared to contain analyte. 7.4 Instrument Sensitivity The smallest standard amount injected during the chromatographic run was 0.5 ng/mL which is equivalent to 0.2 ng/g of PFOS, FOSA and PFOA in matrix. 7.5 Description of Instrument and Operating Conditions A Micromass Quattro Ultima LC/MS/MS coupled to a Hewlet Packard HPLC system was used. Data acquisition and processing were performed using Masslynx 3.4 software. Detailed operating conditions are listed below: Instrument : Micromass Quattro Ultima ELECTROSPRAY ION SOURCE: Capillary: 2.0 kV Hexapole 1: 0 V Aperture 1: 0 V Hexapole 2: 0 V Source Block Temp.: 100C Desolvation Temp.: 400C ANALYmR: LM Res 1: 10.5 V HMRes 1: 10.5 V IEnergy 1: 1.0 V Entrance: -2 V Exit: 2 V LM Res 2: 12.0 V HM Res 2: 12.0 V IEnergy 2: 2.0 V Multiplier: 650 V GAS FLOWS AND PRESSURE: Desolvation N2Flow Rate::-700 L/hr Nebuliser Nz Flow Rate: -150 L/hr Gas Cell Pressure: -0.003 1mbar Computer: Software: HPLC Equipment: HPLC Column: COMPAQ Professional Workstation AP200 Microsoft Windows NT: Version 4 Build 1381: Service Pack 5 Micromass Limited: Masslynx 3.4 Build 004 Hewlett Packard (HP)Series 1100 HP Binary Pump HP Autosampler HP VacuumDegasser HP Column Oven Genesis C-8, 5 cm x 2.1 mni i.d. x 4 p Column Temperature: 35C Mobile Phase (A) : 2 mM Ammonium Acetate iin Type I Water Centre AnalyticalLaboratories,Inc. Page 17 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Mobile Phase (B) : Methanol Time (min) %A 0.0 60.0 0.4 60.0 1.o 10.0 7.0 10.0 7.5 0.0 9.0 0.0 9.5 60.0 13.5 60.0 14.0 60.0 Total run time = 14 min %B 40.0 40.0 90.0 90.0 100.0 100.0 40.0 40.0 40.0 Flow Rate (mWmin) 0.3 0.3 0.3 0.3 0.3 0.4 0.4 0.4 0.3 Injected Volume: 15 pL Ions monitored : Anal yte PFOA FOSA PFOS Parent ion 413 498 499 Daughter ion 369 78 99 Dwell Csecs) 0.2 0.2 0.2 Coll Energy CeV) 10 40 50 Cone CV) 30 30 30 7.6 Quantitation and Example Calculation Fifteen microliters of sample or calibration standard was injected into the LC/MS/MS. The peak area was measured and the standard curve was generated (using l/x weighted linear regression) by Masslynx software using seven concentrations of standards. The residue concentration in food samples was determined using the following equations: Equation 1 was used to calculate the amount of analyte found (in ng/mL, based on peak area) using the standard curve (linear regression parameters) generated by the Masslynx software program. Equation 1: Analyte found (ng/mL) = &ak area - intercept) slope The component residue concentration (in ng/g) in matrix was determined by using Equation 2. Centre Analytical Laboratories, Inc. Page 18 of 151 BACK TO MAIN Equation 2: Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Analyte found (ng/g) = [analyte found (ng/mL) - average analytefound in blanks (ng/mT.J] x DF x FV (mL) sample weight (g) where DF = dilution factor and FV = final volume Note: the process of subtracting the average analyte found in the blanks was done only for control matrix spikes or if a reagent blank contained residue. For samples fortified with known amounts of analytes prior to extraction, Equation 3 was used to calculate the percent recovery. Equation 3: Recovery (%) = - analyte:found (ng/g) x 100 analyte added (ng/g) An example of a calculation using an actual sample follows: Pork sample Centre ID 0009348 Spk A1 (Set No.: 011501A Pork), fortified at 2.5 ng/g of PFOS, FOSA and PFOA: (Calculation is using values from FOSA) Where: peak area intercept slope final volume dilution factor sample weight ng/g added (fort level) amt found in control 149454 2524.48 25194.9 2Id. 1 5 ; 2.5 nglg 0 (NQ = Not quantifiable; assumed to be 0) From equation 1: Analyte found (ng/mL) = 1149484 - 2524.481 25 194.9 = 5.83 nghd. Centre Analytical Laboratories, Inc. Page 19 of 151 From equation 2: Analyte found (ng/g) BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 = (5.832 - 0) x 1 x 2 5.0 = 2.33 mg/g From equation 3: % Recovery = 2.33 :< 100 2.5 = 93% Note: This example calculation was done using rounded numbers, and therefore may be slightly different from the values shown in the RAW DATA. Any residues found for PFOA were adjusted for PFOA purity (95%). This was done by taking the residue found (ng/g) for PFOA and multiplying by the purity (0.95). Other statistical methods used in analyzing this data were: Standard Deviation = 1i=l n-1 Relative Standard Deviation (RSD or Coeflcient of Variation (CV))= Standard Deviatic3 x 100% Mean 8.0 RESULTS AND DISCUSSION Average fortification recoveries and relative standard deviations were 101% f 9.0% for PFOS, 96% f 13% for FOSA and 104%f 14% for PFOA. A summary of all of the fortification recoveries can be found in Table I. Residues ranging from non-quantifiable levells to 0.852 ppb (ng/g) for PFOS, nonquantifiable levels for FOSA, and non-quantifiable to 2.35 ng/g for PFOA (one value of 14.7 ppb (ng/g) was found for PFOA but was considered suspect) were found in the food samples. The residues found in all of the samples are detailed in Table 11. Centre Analytical Laboratories, Inc. Page 20 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Typical calibration curves and chromatograms representing standards, controls, fortifications, and samples are depicted in Figures 1-23. In this study, analytes were not quantified below 0.5 ng/g. If lower detection levels are required, a more definitive study, including extraction blanks in addition to matrix blanks, could be done. Initially the PFOA standard and fortification solutions were not corrected for purity, as purity information had not been received at the time. Purity data was received after residue analyses were completed and the residues found were then corrected for PFOA purity. All ng/g residues of PFOA in this report have been adjusted to reflect the purity of the PFOA used in this study. There was some PFOA contamination from the lab while performing the extraction for eggs. It was believed to be coming from the (extractionsolvent itself. Using a different brand and lot number of ACN alleviated the problem because upon reanalysis, there were no quantifiable residues of PFOA in the egg samples. 9.0 CIRCUMSTANCES THAT MAY HAVE AFFECTED THE DATA There were no circumstances that may have afkcted the quality or integrity of the data. 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 792.195 (b). Retained samples of reference substances are arlchived by the sponsor. Centre Analytical Laboratories, Inc. Page 21 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 11.0 TABLES Centre Analytical Laboratories, Inc. Page 22 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Table I. Summary of PFOS, FOSA and PFOA Fortification Recoveries Centre ID Extraction Sample Description Date Analysis Fort Level Recovery (%) Date (ng/g) PFOS FOSA PFOA 0009341 Spk A1 0009341 Spk B1 0009341 Spk A2 0009341 Spk B2 0009341 Spk A3 0009341 Spk B3 0009341 Spk A4 0009341 Spk B4 0009348 Spk AI 0009348 Spk B 1 0009348 Spk A2 0009348 Spk B2 0009348 Spk A3 0009348 Spk B3 0009348 Spk A4 0009348 Spk B4 0009344 Spk A1 0009344 Spk B 1 0009344 Spk A1 0009344 Spk B 1 0009344 Spk A2 0009344 Spk B2 0009344 Spk A3 0009344 Spk B3 0009344 Spk A4 0009344 Spk B4 0009345 Spk A1 0009345 Spk B 1 0009345 Spk A2 0009345 Spk B2 0009345 Spk A2 0009345 Spk B2 0009345 Spk A3 0009345 Spk B3 0009345 Spk A4 0009345 Spk B4 Chicken Chicken Chicken Chicken Chicken Chicken Chicken Chicken Pork Pork Pork Pork Pork Pork Pork Pork Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Fish Fish Fish Fish Fish Fish Fish Fish Fish Fish 01/11/01 0 1/11-12/01 2.5 01/11/01 0 MI-12/01 10 01/11/01 01/12/01 2.5 01/11/01 101/12/01 10 01/12/01 101/15/01 2.5 01/12101 (31/15/01 10 0I/12/01 01/15/01 2.5 01/12/01 O1/15/01 10 01/15/01 0I / 15-16/01 2.5 01/15/01 0I/15- 16/01 10 01/15/01 01/16/01 2.5 01/15/01 01/16/01 10 01/16/01 01/16/01 2.5 01/16/01 01/16/01 10 01/16101 01/16-17/01 2.5 0I/16101 01/16-17/01 10 01/17/01 01/17-18/01 2.5 01/17/01 01/17-18/01 10 01/17101 01/18/01 2.5 01/17/01 01/18/01 10 0I/17101 01/18/01 2.5 01/17/01 01/18/01 10 01/18/01 01.118-19/01 2.5 01/18/01 01.118-19101 10 01118101 011/19/01 2.5 01/18/01 011/19/01 10 01I19101 01/19-20/01 2.5 01/19/01 0lJ19-20/01 10 01/19/01 01/20/01 2.5 01/19/01 01/20/01 10 01/30/01 01/31/01 2.5 01/30/01 01/31/O 1 10 01/22/01 01~'22-23/01 2.5 01/22/01 01122-23/01 10 01/22/01 01/23/01 2.5 01/22/01 01/23/01 10 104 93 94 89 84 87 110 94 98 91 89 91 108 107 106 106 105 111 106 105 107 97 90 98 105 93 116 99 86 110 98 87 105 84 76 95 98 93 106 81 78 91 90 86 93 82 79 92 102 100 * 95 92 * 97 95 113 106 114 95 91 87 99 94 102 89 77 89 104 95 96 91 84 84 100 68 105 87 56 87 86 * 85 86 * 86 126 110 111 76 111 95 65 101 106 73 103 89 62 89 * = Rejected due to an unacceptablerecovery. Replaced .withre-extracted data. - = Not Analyzed For Centre Analytical Laboratories, Inc. Page 23 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Table I continued: Summary of PFOS, FOSA and PFOA Fortification Recoveries Extraction Centre ID Sample Description Date 0009342 Spk A1 0009342 Spk B 1 0009342 Spk A2 0009342 Spk B2 0009342 Spk A3 0009342 Spk B3 0009342 Spk A4 0009342 Spk B4 0009346 Spk A1 Egg Egg Egg Egg Egg Egg Egg Egg Green Bean 01/23/01 0 112310 1 0 1/23/01 0 112310 1 0 112410 1 01/24/01 01124101 0 112410 1 02/05/0 1 0009346 Spk B 1 Green Bean 02/05/0 1 0009346 Spk A2 Green Bean 02/06/01 0009346 Spk B2 0009346 Spk A3 0009346 Spk B3 Green Bean Green Bean Green Bean 02/06/0 1 02/08/01 02/08/0 1 0009346 Spk A4 0009346 Spk B4 0009349 Spk A1 0009349 Spk B 1 0009349 Spk A2 0009349 Spk B2 0009349 Spk A3 0009349 Spk B3 0009349 Spk A4 Green Bean Green Bean Milk Milk Milk Milk Milk Milk Milk 02/08/0 1 02/08/0 1 02/07/0 1 02/07/0 1 02/07/0 1 02/07/0 1 02108/0 1 02/08/0 1 02/08/0 1 0009349 Spk B4 0009347 Spk A1 0009347 Spk B 1 0009347 Spk A2 0009347 Spk B2 0009347 Spk A3 0009347 Spk B3 0009347 Spk A4 0009347 Spk B4 Milk Apple Apple Apple Apple Apple Apple Apple Apple 0210810 1 02/09101 02/09/0 1 02/12/01 02112101 02112101 02112101 02112101 0211 210 1 Analysis Fort Level Date (nglg) 0Y./23-24/01 2.5 01.123-24101 10 O1/24/01 2.5 01/24/01 10 01124-25/01 2.5 01/24-25101 10 01/25/01 2.5 01/25/01 10 02/05/01 2.5 0210510 1 10 02/07/01 2.5 02/07/0 1 10 02/09/0 1 2.5 O2l09lO 1 10 02112101 2.5 02/12/0 1 10 02/09-10101 2.5 02109-10101 10 0121 1 010 1 2.5 0'2110101 10 02110101 2.5 02110101 10 02,'lO-11/01 2.5 02,'lO-11/01 10 02/12/0 1 2.5 02112101 10 0211 310 1 2.5 02113101 10 02113-14101 2.5 02113-14101 10 0:2/14/01 2.5 021 1410 1 10 Recovery (%) PFOS FOSA PFOA 104 96 84 98 80 67 117 106 78 101 98 77 115 109 105 92 90 85 117 111 90 102 97 82 98 114 129 106 106 128 92 99 124 111 103 130 99 100 125 94 97 125 83 85 103 96 95 112 99 103 129 91 90 128 114 101 123 90 91 123 101 103 126 89 91 129 113 103 124 93 94 123 113 110 122 105 104 119 111 107 108 100 102 101 109 105 110 98 96 100 108 101 102 99 96 99 Centre Analytical Laboratories, Inc. Page 24 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Table I continued: Summary of PFOS, FOSA and PFOA Fortification Recoveries 0009343 Spk A1 0009343 Spk B1 0009343 Spk A2 0009343 Spk B2 0009343 Spk A3 0009343 Spk B3 0009343 Spk A4 0009343 Spk B4 0100006 Spk A1 0100006 Spk B1 01oooO6 Spk A2 01oooO6 Spk B2 0100006 Spk A3 0100006 Spk B3 0100006 Spk A4 0100006 Spk B4 0009342 Spk A1 0009342 Spk B 1 0009342 Spk A2 0009342 Spk B2 0009342 Spk A3 0009342 Spk B3 0009342 Spk A4 0009342 Spk B4 0009342 Spk A1 0009342 Spk B 1 0009342 Spk A2 0009342 Spk B2 0009342 Spk A3 0009342 Spk B3 0009342 Spk A4 0009342 Spk B4 Bread Bread Bread Bread Bread Bread Bread Bread Ground Beef Ground Beef Ground Beef Ground Beef Ground Beef Ground Beef Ground Beef Ground Beef Egg Egg Egg Egg Egg Egg Egg Egg Egg Egg Egg Egg Egg Egg Egg Egg - = Not Analyzed For Extraction Analysis Fort Level Recovery (%) Date (ng/g) PFOS FOSA PFOA 02/08-09/01 Q2/11/01 2.5 87 106 118 02/08-09/01 02/11/01 10 96 96 109 02/08-09/01 02/11/01 2.5 106 101 119 02/08-09/01 02/11/01 10 94 93 106 02/12-13101 02/14/01 2.5 106 103 103 02/12-13/01 02/14/01 10 99 92 97 02/12-13/01 02/14/01 2.5 111 109 106 02/12-13/01 02/14/01 10 102 100 100 02/13/01 02./14-15/01 2.5 104 104 100 02/13/01 02/14-1 9 01 10 103 99 95 02/13101 02/15/01 2.5 111 102 103 02/13101 02/15/01 10 103 98 98 02/14101 02/15/01 2.5 121 123 120 02/14/01 02/15/01 10 106 116 114 02/14/01 02/1516/01 2.5 112 108 105 02/14101 02/15-16/01 10 102 101 102 05/07-08/01 05/08/01 2.5 111 05/07-08/01 05/08/01 10 99 05/07-08/01 05108-09101 2.5 111 0907-08/01 05/08-09/01 10 102 05/08-09/01 O5/09/01 2.5 106 0908-09/01 05/09/01 10 94 05/08-09/01 05,109-10/01 2.5 109 05/08-09/01 05,109-10/01 10 102 OW1 810 1 0'5118/0 1 2.5 110 05/18101 05/18/01 10 99 0 9 18/01 05r'l8-19/01 2.5 102 05/18/01 05118-19/01 10 108 05/20-21/01 0 9 21/01 2.5 96 0920-21/01 05/21/01 10 96 05/20-21/01 05121-22/01 2.5 95 05/20-21101 05121-22/01 10 96 AVERAGE: 101 96 104 STD DEV: 9.0 13 14 RELATIVE STD DEV: 9.0 13 14 n: 88 80 88 Centre Analytical Laboratories, Inc. Page 25 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Table 11. Summary of PFOS, FOSA arid PFOA Residues Found in Samples Centre ID Data Set No. Sponsor ID Store Name Location 0012776 011lOlA Chicken 0012776 Dup 011lOlA Chicken 0012777 011lOlA Chicken 18249 18249 18226 Publix Port St. Lucie, FL Publix Port S t . Lucie, FL Winn-Dixie Port St. Lucie, FL 0012777 Dup 011lOlA Chicken 0012778 011lOlA Chicken 0012778 Dup 011lOlA Chicken 0012779 011lOlA Chicken 0012779 Dup 011lOlA Chicken 18226 18239 18239 18356 18356 Winn-Dixie Port St. Lucie, FL Albertson's Port St. Lucie, FL Albertson's Port St. Lucie, FL Delchamps I1 Mobile, AL Delchamps I1 Mobile, AL 0012780 011lOlA Chicken 0012780 Dup 011lOlA Chicken 18197 18197 Winn-Dixie Columbus, GA Winn-Dixie Columbus, GA 0012781 011lOlB Chicken 0012781 Dup 011lOlB Chicken 18345 18345 Delchamps I Delchamps I Mobile, AL Mobile, AL 0012782 0012782 Dup 0012783 0012783 Dup 0012784 0012784 Dup 0012785 0012785 Dup 0012786 0012786 Dup 0012787 011lOlB Chicken 011lOlB Chicken 011lOlB Chicken 011lOlB Chicken 011lOlB Chicken 011lOlB Chicken 011lOlB Chicken 011lOlB Chicken 011201A Chicken 011201A Chicken 01 1201A Chicken 18209 18209 18335 18335 18323 18323 18312 18312 18301 18301 18187 Piggly Wiggly Piggly Wiggly Bruno's Bruno's Kroger's Kroger's Winn-Dixie Winn-Dixie Haloway Haloway Cub Columbus, GA Columbus, GA Mobile, AL Mobile, AL Decatur, AL Decatur, AL Decatur, AL Decatur, AL Decatur, AL Decatur, AL Columbus, GA 0012787 Dup 0012788 0012788 Dup 0012789 0012789 Dup 0012790 0012790 Dup 0012791 0012791 Dup 011201A Chicken 01 1201A Chicken 01 1201A Chicken 01 1201A Chicken 011201A Chicken 01 1201A Chicken 01 1201A Chicken 011201B Chicken 011201B Chicken 18187 18174 18174 18163 18163 18283 18283 18260 18260 Cub B i-Lo Bi-Lo Save-a-Lot Save-a-Lot Albertson's Albertson's Food World Food World Columbus, GA Cleveland, TN Cleveland, TN Cleveland, TN Cleveland, TN Pensacola, FL Pensacola, FL Pensacola, FL Pensacola, FL Matrix Chicken Chicken Chicken Chicken Chicken Chicken Chicken Chicken Chicken Chicken Chicken Chicken Chicken Chicken Chicken Chicken Chicken Chicken Chicken Chicken Chicken Chicken Chicken Chicken Chicken Chicken Chicken Chicken Chicken Chicken Chicken Chicken Residue Found (ng/g)* PFOS FOSA PFOA NQ = Not Quantifiable. The residue detected, if any, was below the limit of the study which is 0.5 ng/g. *Value is based on wet weight (5.0 g sample processed as received) ** Values rejected based on unacceptable recoveries. Data has been replaced with either reanalysis or re-extraction values. = Results rejected based on apparent contamination. Data has been replaced with re-extracted values. - = Not analyzed for Centre Analytical Laboratories, Inc. Page 26 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Table I1 continued: Summary of PFOS,,FOSA and PFOA Residues Found in Samples Centre ID Data Set No. Sponsor ID Store Name Location 0012792 01 1201B Chicken 18151 Watson's Cleveland, TN 0012792 Dup 0012793 0012793 Dup 0012794 0012794 Dup 0012795 0012795 Dup 0012796 0012796 Dup 0012797 0012797 Dup 0012798 011201B Chicken 011201B Chicken 01 1201B Chicken 01 1501A Pork 01 1501A Pork 01 1501A Pork 01 1501A Pork 01 1501A Pork 01 1501A Pork 01 1501A Pork 01 1501A Pork 01 1501A Pork 18151 18272 18272 18250 18250 18227 18227 18238 18238 18357 18357 18198 Watson's Cleveland, TN Food World I1 Pensacola, FL Food World I1 Pensacola, FL Publix Port St. Lucie, FL Publix Port St. Lucie, FL Winn-Dixie Port St. Lucie, FL Winn-Dixie Port St. Lucie, FL Albertson's Port St. Lucie, FL Albertson's Port St. Lucie, FL Delchamps 11 Mobile, AL Delchamps I1 Mobile, AL Winn-Dixie Columbus, GA 0012798 Dup 0012799 0012799 Dup 0012800 0012800 Dup 0012801 0012801 Dup 0012802 0012802 Dup 0012803 0012803 Dup 01 1501A Pork 01 1501B Pork 011501B Pork 01 1501B Pork 01 1501B Pork 01 1501B Pork 011501B Pork 01 1501B Pork 011501B Pork 011501B Pork 011501B Pork 18198 18346 18346 18210 18210 18334 18334 18322 18322 18311 18311 Winn-Dixie Delchamps I Delchamps I Piggly Wiggly Piggly Wiggly Bruno's Bruno's Kroger's Kroger's Winn-Dixie Winn-Dixie Columbus, GA Mobile, AL Mobile, AL Columbus, GA Clolumbus, GA Mobile, AL Mobile, AL Decatur, AL Decatur, AL Decatur, AL Decatur, AL 0012804 0012804 Dup 0012805 0012805 Dup 0012806 0012806 Dup 0012807 0012807 Dup 01 1601A Pork 011601A Pork 01 1601A Pork 01 1601A Pork 011601A Pork 01 1601A Pork 011601A Pork 011601APork 18299 18299 18186 18186 18175 18175 18164 18164 Haloway Haloway Cub Cub B i-Lo Bi-Lo Save-a-Lot Save-a-Lot .Decatur, AL Decatur, AL Columbus, GA Columbus, GA Cleveland, TN Cleveland, TN Cleveland, TN Cleveland, TN Matrix Chicken Chicken Chicken Chicken Pork Pork Pork Pork Pork Pork Pork Pork Pork Pork Pork Pork Pork Pork Pork Pork Pork Pork Pork Pork Pork Pork Pork Pork Pork Pork Pork Pork Residue Found (ng/g)* PFOS FOSA PFOA NQ = Not Quantifiable. The residue detected, if any, was below the limit of the study which is 0.5 ng/g. *Value is based on wet weight (5.0 g sample processed as received) ** Values rejected based on unacceptable recoveries. Data has been replaced with either reanalysis or re-extraction values. AA = Results rejected based on apparent contamination. Data has been replaced with re-extracted values. - = Not analyzed for Centre Analytical Laboratories, Inc. Page 27 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Table I1 continued: Summary of PFOS, FOSA and PFOA Residues Found in Samples Centre ID Data Set No. Sponsor ID Store Name Location 0012808 0012808 Dup 0012809 01 1601A Pork 01 1601A Pork 011601B Pork 18282 18282 18261 Albertson's Albertson's Food World Pensacola, FL Pensacola, FL Pensacola, FL 0012809 Dup 0012810 0012810 Dup 011601B Pork 011601B Pork 011601B Pork 18261 18152 18152 Food World Watson's Watson's Pensacola, FL Cleveland, TN Cleveland, TN 0012811 011601BPork 0012811 Dup 011601BPork 18271 Food World I1 Pensacola,FL 18271 Food World I1 Pensacola,FL 0012812 011701AHot Dog 0012812 Dup 011701A Hot Dog 0012813 011701A Hot Dog 0012813 Dup 01 1701A Hot Dog 18251 18251 18228 18228 Publix Publix Winn-Dixie Winn-Dixie Port St. Lucie, FL Port St. Lucie, FL Port St. Lucie, FL Port St. Lucie, FL 0012814 011701AHot Dog 18240 Albertson's Port St. Lucie, FL 0012814 Dup 011701A Hot Dog 18240 Albertson's Port St. Lucie, FL 0012815 01 1701A Hot Dog 18358 Delchamps I1 Mobile, AL 0012815 Dup 011701AHot Dog 0012816 011701AHot Dog 18358 18199 Delchamps I1 Mobile, AL Winn-Dixie Columbus, GA 0012816 Dup 011701AHot Dog 18199 0012812 011701AR Hot Dog 18251 Winn-Dixie Columbus, GA Publix Port St. Lucie, FL 0012812 Dup 011701AR Hot Dog 18251 0012813 011701AR Hot Dog 18228 Publix Port St. Lucie, FL Winn-Dixie Port St. Lucie, FL 0012813 Dup 011701AR Hot Dog 18228 Winn-Dixie Port St. Lucie, FL 0012814 011701AR Hot Dog 18240 0012814 Dup 011701ARHot Dog 18240 Albertson's Port St. Lucie, FL Albertson's Port St. Lucie, FL 0012815 011701ARHot Dog 0012815 Dup 011701AR Hot Dog 18358 18358 Delchamps I1 Delchamps I1 Mobile, AL Mobile, AL 0012816 011701AR Hot Dog 18199 Winn-Dixie Columbus, GA 0012816 Dup 011701AR Hot Dog 18199 0012817 011701B Hot Dog 18347 Winn-Dixie Columbus, GA Delchamps I Mobile, AL 0012817 Dup 011701B Hot Dog 18347 Delchamps I Mobile, AL 0012818 011701B Hot Dog 0012818 Dup 011701BHot Dog 18211 Piggly Wiggly Columbus, GA 18211 Piggly Wiggly Columbus, GA Matrix Pork Pork Pork Pork Pork Pork Pork Pork Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Residue Found (ng/g)* PFOS FOSA PFOA NQ NQ NQ NQ NQ NQ NQ N*Q* ** ** ** ** ** ** ** ** ** NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ = Not Quantifiable. The residue detected, if any, was below the limit of the study which is 0.5 ng/g. *Value is based on wet weight (5.0 g sample processed as received) ** Values rejected based onunacceptable-recoveries. Data has been replaced with either reanalysisor re-extraction values. = Results rejected based on apparent contamination. Data has been replaced with re-extracted values. - = Not analyzed for Centre Analytical Laboratories, Inc. Page 28 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Table I1 continued: Summary of PFOS,,FOSA and PFOA Residues Found in Samples Centre ID Data Set No. Sponsor ID Store Name Location 0012819 011701B Hot Dog 0012819 Dup 011701B Hot Dog 0012820 01 1701B Hot Dog 0012820 Dup 011701B Hot Dog 0012821 01 1701B Hot Dog 0012821 Dup 01 1701B Hot Dog 0012822 011801A Hot Dog 0012822 Dup 01 1801A Hot Dog 0012823 01 1801A Hot Dog 0012823 Dup 011801AHotDog 0012824 01 1801A Hot Dog 0012824Dup 011801A Hot Dog 0012825 01 1801A Hot Dog 0012825 Dup 01 1801A Hot Dog 0012826 01 1801A Hot Dog 0012826 Dup 01 1801A Hot Dog 0012827 01 1801B Hot Dog 0012827 Dup 01 1801B Hot Dog 0012828 01 1801B Hot Dog 0012828 Dup 01 1801B Hot Dog 0012829 011801B Hot Dog 0012829 Dup 01 1801B Hot Dog 0012830 01 1901A Fish 0012830 Dup 011901A Fish 0012831 01 1901A Fish 0012831 Dup 011901AFish 0012832 01 1901A Fish 0012832 Dup 01 1901A Fish 0012833 01 1901A Fish 0012833 Dup 01 1901A Fish 0012834 01 1901A Fish 0012834 Dup 011901AFish 18336 18336 18324 18324 18313 18313 18302 18302 18188 18188 18176 18176 18165 18165 18285 18285 18263 18263 18153 18153 18273 18273 18252 18252 18229 18229 1824 1 1824 1 18359 18359 18200 18200 Bruno's Mobile, AL Bruno's Mobile, AL Kroger's Decatur, AL Kroger's Decatur, AL Winn-Dixie Decatur, AL Winn-Dixie Decatur, AL Haloway Decatur, AL Haloway Decatur, AL Cub Columbus, GA Cub Columbus, GA B i-Lo Cleveland, TN Bi-Lo Cleveland, TN Save-a-Lot Cleveland, TN Save-a-Lot Cleveland,TN Albertson's I'ensacola, FL Albertson's I'ensacola, FL Food World Pensacola, FL Food World Pensacola,FL Watson's Cleveland,TN Watson's Cleveland,TN Food World I1 F'ensacola, FL Food World I1 F'ensacola,FL Publix Poirt St. Lucie, FL Publix Port St. Lucie, FL Winn-Dixie Port St. Lucie, FL Winn-Dixie Port St. Lucie, FL Albertson's Port St. Lucie, FL Albertson's Port St. Lucie, FL Delchamps I1 Mobile, AL Delchamps I1 Mobile, AL Winn-Dixie Columbus, GA Winn-Dixie Columbus, GA Matrix Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Hot Dog Fish Fish Fish Fish Fish Fish Fish Fish Fish Fish Residue Found (ng/g)* PFOS FOSA PFOA NQ = Not Quantifiable. The residue detected, if any, was below the limit of the study which is 0.5 ng/g. *Value is based on wet weight (5.0 g sample processed as received) ** Values rejected based on unacceptable recoveries. Data has been replaced with either reanalysisor re-extractionvalues. = Results rejected based on apparent contamination. Data has been replaced with re-extracted values. - = Not analyzed for Centre Analytical Laboratories, Inc. Page 29 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Table I1 continued: Summary of PFOS, FOSA and PFOA Residues Found in Samples Centre ID 0012835 0012835 Dup 0012836 0012836 Dup 0012837 0012837 Dup 0012838 0012838 Dup 0012839 0012839 Dup 0012835 0012835 Dup 0012836 0012836 Dup 0012837 0012837 Dup 0012838 0012838 Dup 0012839 0012839 Dup 0012840 0012840 Dup 0012841 0012841 Dup 00 12842 0012842 Dup 0012843 0012843 Dup 0012844 0012844 Dup 0012845 0012845 Dup Data Set No. 01 1901B Fish 01 1901B Fish 01 1901B Fish 01 1901B Fish 01 1901B Fish 01 1901B Fish 01 1901B Fish 01 1901B Fish 011901B Fish 01 1901B Fish 013001A Fish 013001A Fish 013001A Fish 013001A Fish 013001AFish 013001A Fish 013001A Fish 013001A Fish 013001A Fish 013001A Fish 012201A Fish 012201A Fish 012201A Fish 012201A Fish 012201A Fish 012201A Fish 012201A Fish 012201A Fish 012201A Fish 012201A Fish 012201B Fish 012201B Fish Sponsor ID Store Name 18348 Delchamps 18348 Delchamps 18212 Piggly Wiggly 18212 Piggly Wiggly 18337 18337 18325 18325 18314 Bruno's Bruno's Kroger's Kroger 's Winn-Dixie 18314 Winn-Dixie 18348 18348 18212 Delchamps Delchamps Piggly Wiggly 18212 18337 18337 18325 18325 18314 Piggly Wiggly Bruno's Bruno's Kroger's Kroger's Winn-Dixie 18314 Winn-Dixie 18303 18303 18177 18177 Halo way Haloway Bi-Lo Bi-Lo 18166 18166 Save-a-Lot Save-a-Lot 18284 18284 Albertson's Albertson's 18262 18262 18154 Food World Food World Watson's 18154 Watson's Location Mobile, AL Mobile, AL Columbus, GA Columbus, GA Mobile, AL Mobile, AL Decatur, AL Decatur, AL Decatur, AL Decatur, AL Mobile, AL Mobile, AL Columbus, GA Clolumbus, GA Mobile, AL Mobile, AL Decatur, AL Decatur, AL Decatur, AL Decatur, AL :Decatur, AL IDecatur, AL Cleveland, TN Cleveland, TN Cleveland, TN Cleveland, TN Pensacola, FL Pensacola, FL Pensacola, FL Plensacola, FL Cleveland, TN Clleveland, TN Matrix Fish Fish Fish Fish Fish Fish Fish Fish Fish Fish Fish Fish Fish Fish Fish Fish Fish Fish Fish Fish Fish Fish Fish Fish Fish Fish Fish Fish Fish Fish Fish Fish Residue Found (ng/g)* PFOS FOSA ** ** ** ** ** ** ** ** ** ** PFOA 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. The residue detected, if any, was below the limit of the study which is 0.5 ng/g. *Value is based on wet weight (5.0 g sample processed as received) ** Values rejected based on unacceptable recoveries. Data has been replaced with either reanalysisor re-extractionvalues. Ah = Results rejected based on apparent contamination. Data has been replaced with re-extracted values. - = Not analyzed for Centre Analytical Laboratories, Inc. Page 30 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Table I1 continued: Summary of PFOS, FOSA and PFOA Residues Found in Samples Centre ID 0012846 0012846 Dup 0012903 0012903 Dup 0012904 0012904 Dup 00 12905 0012905 Dup 00 12906 00 12906 Dup 00 12907 0012907 Dup 0012908 0012908 Dup 00 12909 0012909 Dup 0012910 0012910 Dup 0012911 0012911 Dup 0012912 0012912 Dup 0012913 0012913 Dup 0012914 0012914 Dup 0012915 0012915 Dup 00129 16 0012916 Dup 0012917 0012917 Dup Data Set No. 012201B Fish 012201B Fish 020501A Bean 020501A Bean 020501A Bean 020501A Bean 020501A Bean 020501A Bean 020501A Bean 020501A Bean 020501A Bean 020501A Bean 020601C Bean 020601C Bean 020601C Bean 020601C Bean 020601C Bean 020601C Bean 020601C Bean 020601C Bean 020601C Bean 020601C Bean 020801C Bean 020801C Bean 020801C Bean 020801C Bean 020801C Bean 020801C Bean 020801C Bean 020801C Bean 020801C Bean 020801C Bean Residue Found (ng/g)* Sponsor ID Store Name Location Matrix PFOS FOSA PFOA 18274 18274 18256 18256 Food World I1 Pensacola, FL Fish Food World I1 Pensacola, FL Fish Publix Port St. Lucie, FL Green Bean Publix Port St. Lucie, FL Green Bean 18234 18234 18245 18245 Winn-Dixie Winn-Dixie Albertson's Albertson's Port St. Lucie, FL Green Bean Port St. Lucie, FL Green Bean Port St. Lucie, FL Green Bean Port St. Lucie, FL Green Bean 18363 Delchamps I1 Mobile, AL Green Bean 18363 Delchamps I1 Mobile, AL Green Bean 18204 18204 Winn-Dixie Columbus, GA Green Bean Winn-Dixie Columbus, GA Green Bean 18352 Delchamps I Mobile, AL Green Bean 18352 Delchamps I Mobile, AL Green Bean 18216 Piggly Wiggly Columbus, GA Green Bean 18216 Piggly Wiggly Columbus, GA Green Bean 18341 Bruno's Mobile, AL Green Bean 18341 Bruno's Mobile, AL Green Bean 18329 Kroger's Decatur, AL Green Bean 18329 18318 18318 Kroger's Winn-Dixie Winn-Dixie Decatur, AL Decatur, AL Decatur, AL Green Bean Green Bean Green Bean 18307 Haloway Decatur, AL Green Bean 18307 Haloway Decatur, AL Green Bean 18193 18193 18182 Cub Cub Bi-Lo Columbus, GA Columbus, GA Cleveland, TN Green Bean Green Bean Green Bean 18182 18289 Bi-Lo Albertson's Cleveland, TN Green Bean Pensacola, FL Green Bean 18289 18267 18267 Albertson's Food World Food World Pensacola, FL Pensacola, FL Pensacola, FL Green Bean Green Bean Green Bean NQ = Not Quantifiable. The residue detected, if any, was below the limit of the study which is 0.5 ng/g. *Value is based on wet weight (5.0 g sample processed as received) ** Values rejected based on unacceptable recoveries. Data has been replaced with either reanalysis or re-extraction values. AA = Results rejected based on apparent contamination. Data has been replaced with re-extracted values. - = Not analyzed for Centre Analytical Laboratories,Inc. Page 31 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Table I1 continued: Summary of PFOS,,FOSA and PFOA Residues Found in Samples Centre ID 0012918 0012918 Dup 0012919 0012919 Dup 0012867 0012867 Dup 0012868 0012868 Dup 0012869 0012869 Dup 0012870 0012870 Dup 0012871 0012871 Dup 0012872 0012872 Dup 0012873 0012873 Dup 0012874 0012874 Dup 0012875 0012875 Dup 0012876 0012876 Dup 0012877 0012877 Dup 0012878 0012878 Dup 0012879 0012879 Dup 0012880 0012880 Dup Data Set No. 020801D Bean 020801D Bean 020801D Bean 020801D Bean 020701A Milk 020701A Milk 020701A Milk 020701A Milk 020701A Milk 020701A Milk 020701A Milk 020701A Milk 020701A Milk 020701A Milk 020701B Milk 020701B Milk 020701B Milk 020701B Milk 020701B Milk 020701B Milk 020701B Milk 020701B Milk 020701B Milk 020701B Milk 020801A Milk 020801A Milk 020801A Milk 020801A Milk 020801A Milk 020801A Milk 020801A Milk 020801A Milk Sponsor ID Store Name Location Matrix 36868 36868 Watson's Watson's Cleveland,TN Green Bean Cleveland,TN Green Bean 18278 Food World I1 :Pensacola, FL Green Bean 18278 Food World I1 Pensacola, FL Green Bean 18254 Publix Port St. Lucie, FL Milk 18254 Publix Port St. Lucie, FL Milk 18231 Winn-Dixie Port St. Lucie, FL Milk 18231 Winn-Dixie Port St. Lucie, FL Milk 18243 Albertson's Port St. Lucie, FL Milk 18243 Albertson's Port St. Lucie, FL Milk 18361 Delchamps I1 Mobile, AL Milk 18361 Delchamps I1 Mobile, AL Milk 18202 Winn-Dixie Columbus, GA Milk 18202 Winn-Dixie Columbus, GA Milk 18350 Delchamps I Mobile, AL Milk 18350 Delchamps I Mobile, AL Milk 18214 Piggly Wiggly Columbus, GA Milk 18214 Piggly Wiggly Clolumbus, GA Milk 18339 Bruno's Mobile, AL Milk 18339 Bruno's Mobile, AL Milk 18327 Kroger's Decatur, AL Milk 18327 Kroger's Decatur, AL Milk 18316 Winn-Dixie Decatur, AL Milk 18316 Winn-Dixie Decatur, AL Milk 18305 Haloway Decatur, AL Milk 18305 Haloway Decatur, AL Milk 18190 Cub Columbus, GA Milk 18190 Cub Columbus, GA Milk 18179 18179 Bi-Lo Bi-Lo Cleveland,TN Milk Cleveland,TN Milk 18168 Save-a-Lot Cleveland,TN Milk 18168 Save-a-Lot Cleveland,TN Milk Residue Found (ng/g)* PFOS FOSA PFOA NQ = Not Quantifiable. The residue detected, if any, was below the limit of the study which is 0.5 ng/g. *Value is based on wet weight (5.0 g sample processed as received) ** Values rejected based on unacceptable recoveries. Data has beten replaced with either reanalysisor re-extractionvalues. AA = Results rejected based on apparent contamination. Data has been replaced with re-extracted values. - = Not analyzed for Centre Analytical Laboratories, Inc. Page 32 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Table I1 continued: Summary of PFOS, FOSA and PFOA Residues Found in Samples Centre ID 0012881 0012881 Dup 0012882 0012882 Dup 0012883 0012883 Dup 0012884 0012884 Dup 0012926 0012926 Dup 0012927 0012927 Dup 0012928 0012928 Dup 0012929 0012929 Dup 0012930 0012930 Dup 0012931 0012931 Dup 0012932 0012932 Dup 0012933 0012933 Dup 0012934 0012934 Dup 0012935 0012935 Dup 0012936 0012936 Dup 0012937 0012937 Dup Data Set No. 020801A Milk 020801A Milk 020801B Milk 020801B Milk 020801B Milk 020801B Milk 020801B Milk 020801B Milk 020901C Apple 020901C Apple 020901C Apple 020901C Apple 020901C Apple 020901C Apple 020901C Apple 020901C Apple 020901C Apple 020901C Apple 021201A Apple 021201A Apple 021201A Apple 021201A Apple 021201A Apple 021201A Apple 021201A Apple 021201A Apple 021201A Apple 021201A Apple 021201B Apple 021201B Apple 021201B Apple 021201B Apple Sponsor ID Store Name Location 18287 Albertson's Pensacola,FL 18287 Albertson's Pensacola,FL 18265 Food World Pensacola,FL 18265 Food World Pensacola,FL 18156 Watson's Cleveland, TN 18156 Watson's Cleveland,TN 18276 Food World I1 Pensacola,FL 18276 Food World I1 Pensacola,FL 18205 Winn-Dixie Columbus, GA 18205 18257 18257 Winn-Dixie Columbus, GA Publix Port St. Lucie, FL Publix Port St. Lucie, FL 18235 18235 Winn-Dixie Port St. Lucie, FL Winn-Dixie Polrt St. Lucie, FL 18246 18246 18364 Albertson's Port St. Lucie, FL Albertson's Port St. Lucie, FL Delchamps I1 Mobile, AL 18364 Delchamps I1 Mobile, AL 18353 Delchamps I Mobile, AL 18353 Delchamps I Mobile, AL 18217 18217 18342 Piggly Wiggly Columbus, GA Piggly Wiggly Columbus, GA Bruno's Mobile, AL 18342 Bruno's Mobile, AL 18330 Kroger's Decatur, AL 18330 Kroger's Decatur, AL 18319 Winn-Dixie Decatur, AL 18319 Winn-Dixie IDecatur, AL 18308 Haloway IDecatur, AL 18308 Haloway IDecatur, AL 18194 Cub Columbus, GA 18194 Cub Clolumbus, GA Matrix Residue Found (ng/g)* PFOS FOSA PFOA NQ = Not Quantifiable. The residue detected, if any, was below the limit of the study which is 0.5 ng/g. *Value is based on wet weight (5.0 g sample processed as received) ** Values rejected based on unacceptable recoveries. Data has been replaced with either reanalysisor re-extraction values. = Results rejected based on apparent contamination. Data has been replaced with re-extracted values. - = Not analyzed for Centre Analytical Laboratories, Inc. Page 33 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Table I1 continued: Summary of PFOS,,FOSA and PFOA Residues Found in Samples Centre ID 0012938 0012938 Dup 0012939 0012939 Dup 0012940 0012940 Dup 0012941 0012941 Dup 0012942 0012942 Dup 0012943 0012943 Dup 0012885 0012885 Dup 0012886 0012886 Dup 0012887 0012887 Dup 0012888 0012888 Dup 0012889 0012889 Dup 0012890 0012890 Dup 0012891 0012891 Dup 0012892 0012892 Dup 0012893 0012893 Dup 0012894 0012894 Dup Data Set No. 021201B Apple 021201B Apple 021201B Apple 021201B Apple 021201B Apple 021201B Apple 021201C Apple 021201C Apple 021201C Apple 021201C Apple 021201C Apple 021201C Apple 020901A Bread 020901A Bread 020901A Bread 020901A Bread 020901A Bread 020901A Bread 020901A Bread 020901A Bread 020901A Bread 020901A Bread 020901B Bread 020901B Bread 020901B Bread 020901B Bread 020901B Bread 020901B Bread 020901B Bread 020901B Bread 020901B Bread 020901B Bread Sponsor ID Store Name Location 18183 18183 18170 18170 18290 18290 18268 18268 18159 18159 18279 18279 18255 18255 18233 18233 18244 18244 18362 18362 18203 18203 18351 18351 18215 18215 Bi-Lo Cleveland, TN Bi-Lo Cleveland,TN Save-a-Lot Cleveland,TN Save-a-Lot Cleveland,TN Albertson's Pensacola, FL Albertson's I'ensacola, FL Food World Pensacola, FL Food World I'ensacola, FL Watson's Cleveland,TN Watson's Cleveland,TN Food World I1 Pensacola, FL Food World I1 Pensacola, FL Publix Port St. Lucie, FL Publix Port St. Lucie, FL Winn-Dixie Port St. Lucie, FL Winn-Dixie Port St. Lucie, FL Albertson's Port St. Lucie, FL Albertson's Port St. Lucie, FL Delchamps I1 Mobile, AL Delchamps I1 Mobile, AL Winn-Dixie Columbus, GA Winn-Dixie Columbus, GA Delchamps I Mobile, AL Delchamps I Mobile, AL Piggly Wiggly Columbus, GA Piggly Wiggly Columbus, GA 18340 18340 18328 18328 18317 18317 Bruno's Bruno's Kroger's Kroger's Winn-Dixie Winn-Dixie Mobile, AL Mobile, AL Ilecatur, AL IJecatur, AL Decatur, AL Decatur, AL Matrix Apple Apple Apple Apple Apple Apple Apple Apple Apple Apple Apple Apple Bread Bread Bread Bread Bread Bread Bread Bread Bread Bread Bread Bread Bread Bread Bread Bread Bread Bread Bread Bread NQ = Not Quantifiable. The residue detected, if any, was below the limit of the study which is 0.5 ng/g. *Value is based on wet weight (5.0 g sample processed as received) ** Values rejected based on unacceptable recoveries. Data has been replaced with either reanalysisor re-extraction values. = Results rejected based on apparent contamination. Data has been replaced with re-extracted values. - = Not analyzed for A = Sample result fell below 0.5 ng/g when adjusting for purity of I'FOA. Centre Analytical Laboratories, Inc. Page 34 of 151 BACK TO MAIN le I1 continuec Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Summary of PFOS, FOSA and PFOA Residues Found in Samples Centre ID 0012895 0012895 Dup 0012896 0012896 Dup 0012897 0012897 Dup 0012898 0012898 Dup 0012899 0012899 Dup 00 12900 0012900 Dup 0012901 0012901 Dup 0012902 0012902 Dup 0012758 0012758 Dup 0012759 0012759 Dup 0012760 0012760 Dup 0012761 0012761 Dup 0012762 0012762 Dup 0012763 0012763 Dup 0012764 0012764 Dup 0012765 00 12765 Dup Data Set No. 021301A Bread 021301A Bread 021301A Bread 021301A Bread 021301A Bread 021301A Bread 021301A Bread 021301A Bread 021301A Bread 021301A Bread 021301B Bread 021301B Bread 021301B Bread 021301B Bread 021301B Bread 021301B Bread 021301C Beef 021301C Beef 021301C Beef 021301C Beef 021301C Beef 021301C Beef 021301C Beef 021301C Beef 021301C Beef 021301C Beef 021301D Beef 021301D Beef 021301D Beef 021301D Beef 021301D Beef 021301D Beef Residue Found (ng/g)* Sponsor ID Store Name Location Matrix PFOS FOSA PFOA 18306 Haloway Decatur, AL Bread NQ 18306 Haloway Decatur, AL Bread NQ 18192 Cub Columbus, GA Bread NQ 18192 Cub Columbus, GA Bread NQ 18181 Bi-Lo Cleveland,TN Bread NQ 18181 Bi-Lo Cleveland,TN Bread NQ 18172 Save-a-Lot Cleveland,TN Bread NQ 18172 Save-a-Lot Cleveland,TN Bread NQ 18288 Albertson's l'ensacola, FL Bread NQ 18288 Albertson's I'ensacola, FL Bread NQ 18266 Food World l'ensacola, FL Bread NQ 18266 Food World I'ensacola, FL Bread NQ 18158 Watson's Cleveland,TN Bread NQ 18158 Watson's Cleveland, TN Bread NQ 18277 Food World I1 I'ensacola, FL Bread NQ 18277 Food World I1 Pensacola,FL Bread NQ 18248 Publix Port St. Lucie, FL Ground Beef NQ 18248 Publix Port St. Lucie, FL Ground Beef NQ 18225 Winn-Dixie Port St. Lucie, FL Ground Beef NQ 18225 Winn-Dixie Port St. Lucie, FL Ground Beef NQ 18237 Albertson's Port St. Lucie, FL Ground Beef NQ 18237 Albertson's Port St. Lucie, FL Ground Beef NQ 18355 Delcharnps I1 Mobile, AL Ground Beef NQ 18355 Delchamps I1 Mobile, AL Ground Beef NQ 18196 Winn-Dixie Columbus, GA Ground Beef NQ 18196 Winn-Dixie Columbus, GA Ground Beef NQ 18344 Delchamps I Mobile, AL Ground Beef 0.570 18344 Delchamps I Mobile, AL Ground Beef 0.587 18208 18208 18333 Piggly Wiggly Columbus, GA Ground Beef NQ Piggly Wiggly Columbus, GA Ground Beef NQ Bruno's Mobile, AL Ground Beef NQ 18333 Bruno's Mobile, AL Ground Beef NQ NQ = Not Quantifiable. The residue detected, if any, was below the limit of the study which is 0.5 ng/g. *Value is based on wet weight (5.0 g sample processed as received) ** Values rejected based on unacceptable recoveries. Data has been replaced with either reanalysisor re-extractionvalues. A = Result is suspect. Apparent contamination. = Results rejected based on apparent contamination. Data has been replaced with re-extracted values. - = Not analyzed for Centre Analytical Laboratories,Inc. Page 35 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Table I1 continued: Summary of PFOS,,FOSA and PFOA Residues Found in Samples Centre ID 0012766 0012766 Dup 0012767 0012767 Dup 0012768 0012768 Dup 0012769 0012769 Dup 0012770 0012770 Dup 0012771 0012771 Dup 0012772 0012772 Dup 0012773 0012773 Dup 00 12774 0012774 Dup 0012775 0012775 Dup 0012847 0012847 Dup 0012848 0012848 Dup 00 12849 0012849 Dup 0012850 0012850 Dup 0012851 0012851 Dup 0012852 0012852 Dup Data Set No. 021301D Beef 021301D Beef 021301D Beef 021301D Beef 021401A Beef 021401ABeef 021401A Beef 021401A Beef 021401A Beef 021401A Beef 021401A Beef 021401A Beef 021401A Beef 021401A Beef 021401B Beef 021401B Beef 021401B Beef 021401B Beef 021401B Beef 021401B Beef 012301A Egg 012301A Egg 012301A Egg 012301A Egg 012301A Egg 012301A Egg 012301A Egg 012301A Egg 012301A Egg 012301A Egg 012301B Egg 012301B Egg Sponsor ID Store Name Location Matrix Residue Found (ng/g)* PFOS FOSA PFOA 18321 18321 18310 18310 18300 18300 18185 18185 18173 18173 18162 18162 18281 1828 1 18259 18259 18161 18161 18270 18270 18253 18253 18230 18230 18242 18242 18360 18360 18201 18201 18349 18349 Kroger's Decatur, AL Ground Beef Kroger's Decatur, AL Ground Beef Winn-Dixie Decatur, AL Ground Beef Winn-Dixie Decatur, AL Ground Beef Haloway Decatur, AL Ground Beef Haloway Decatur, AL Ground Beef Cub Columbus, GA Ground Beef Cub Columbus, GA Ground Beef Bi-Lo Cleveland,TN Ground Beef Bi-Lo Cleveland,TN Ground Beef Save-a-Lot Cleveland,TN Ground Beef Save-a-Lot Cleveland,TN Ground Beef Albertson's l'ensacola, FL Ground Beef Albertson's l'ensacola, FL Ground Beef Food World I'ensacola, FL Ground Beef Food World I'ensacola, FL Ground Beef Watson's Cleveland,TN Ground Beef Watson's Cleveland,TN Ground Beef Food World I1 Pensacola, FL Ground Beef Food World I1 Pensacola,FL Ground Beef Publix Port St. Lucie, FL Egg Publix Port St. Lucie, FL Egg Winn-Dixie Port St. Lucie, FL Egg Winn-Dixie Port St. Lucie, FL Egg Albertson's Po.rt St. Lucie, FL Egg Albertson's Port St. Lucie, FL Egg Delchamps I1 Mobile, AL Egg Delchamps I1 Mobile, AL Egg Winn-Dixie Columbus, GA Egg Winn-Dixie Columbus, GA Egg Delchamps I Mobile, AL Egg Delchamps I Mobile, AL Egg NQ = Not Quantifiable. The residue detected, if any, was below the limit of the study which is 0.5 ng/g. *Value is based on wet weight (5.0 g sample processed as received) ** Values rejected based on unacceptable recoveries. Data has been replaced with either reanalysisor re-extractionvalues. AA = Results rejected based on apparent contamination. Data has been replaced with re-extracted values. - = Not analyzed for Centre Analytical Laboratories, Inc. Page 36 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Table I1 continued: Summary of PFOS.,FOSA and PFOA Residues Found in Samples Centre ID 0012853 0012853 Dup 0012854 0012854 Dup 0012855 0012855 Dup 0012856 0012856 Dup 0012857 0012857 Dup 0012858 0012858 Dup 0012859 0012859 Dup 0012860 0012860 Dup 0012861 0012861 Dup 0012862 0012862 Dup 0012863 0012863 Dup 00 12864 0012864 Dup 0012865 0012865 Dup 0012866 0012866 Dup 0 106429 0106429 Dup 0106430 0106430 Dup Data Set No. 012301B Egg 012301B Egg 012301B Egg 012301B Egg 012301B Egg 012301B Egg 012301B Egg 012301B Egg 012401A Egg 012401A Egg 012401A Egg 012401A Egg 012401A Egg 012401A Egg 012401A Egg 012401A Egg 012401A Egg 012401A Egg 012401B Egg 012401B Egg 012401B Egg 012401B Egg 012401B Egg 012401B Egg 012401B Egg 012401B Egg 012401B Egg 012401B Egg 050801A Egg 050801A Egg 050801A Egg 050801A Egg Sponsor ID Store Name Location 18219 18219 Peggy's Peggy's Port St. Lucie, FL Port St. Lucie, FL 18213 18213 18338 18338 18326 18326 18315 18315 18304 Piggly Wiggly Piggly Wiggly Bruno's Bruno's Kroger's Kroger's W inn-Dixie Winn-Dixie Haloway Columbus, GA Columbus, GA Mobile, AL Mobile, AL Decatur, AL Decatur, AL Decatur, AL Decatur, AL Decatur, AL 18304 18189 18189 Haloway Cub Cub Decatur, AL Columbus, GA Columbus, GA 18178 Bi-Lo Cleveland,TN 18178 Bi-Lo Cleveland, TN 18167 Save-a-Lot Cleveland,TN 18167 Save-a-Lot Cleveland,TN 18286 18286 Albertson's Albertson's Pensacola, FL Pensacola, FL 18264 Food World Pensacola, FL 18264 18296 18296 18155 18155 Food World Farmer's Market Farmer's Market Watson's Watson's Pensacola, FL Mobile, AL Mobile, AL Cleveland, TN Cleveland, TN 18275 18275 21213 21213 21214 21214 Food World I1 Food World I1 Winn-Dixie Winn-Dixie Albertson's Albertson's Pensacola, FL Pensacola, FL Port St. Lucie, FL Port St. Lucie, FL Port St. Lucie, FL Port St. Lucie. FL Matrix Residue Found (ng/g)* PFOS FOSA PFOA Ah All Ah Ah Ah Ah Ah Ah Ah AA Ah AA AA AA AA Ah AA Ah AA Ah AA AA Ah Ah Ah Ah AA AA Ah Ah Ah Ah Ah Ah Ah AA Ah AA AA AA Ah Ah AA AA AA AA AA Ah Ah Ah AA AA AA Ah AA AA NQ Ah NQ Ah NQ AA NQ Ah NQ = Not Quantifiable. The residue detected, if any, was below the limit of the study which is 0.5 nglg. *Value is based on wet weight (5.0 g sample processed as received) ** Values rejected based on unacceptable recoveries. Data has been replaced with either reanalysisor re-extractionvalues. AA = Results rejected based on apparent contamination. Data has been replaced with re-extracted values. - = Not analyzed for Centre Analytical Laboratories, Inc. Page 37 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: 00P-023-057 Table I1 continued: Summary of PFOS,,FOSA and PFOA Residues Found in Samples Centre ID 0106431 0106431 Dup 0106432 0106432 Dup 0 106433 0106433 Dup 0 106434 0106434 Dup 0106435 0106435 Dup 0106436 0106436 Dup 0106437 0106437 Dup 0106438 0106438 Dup 0106439 0106439 Dup 0106440 0106440 Dup 0106441 0106441 Dup 0106442 0106442 Dup 0 106443 0106443 Dup 0106444 0106444 Dup 0 106445 0106445 Dup 0106446 0106446 Dup Data Set No. 050801A Egg 050801A Egg 050801A Egg 050801A Egg 050801A Egg 050801A Egg 050801B Egg 050801B Egg 050801B Egg 050801B Egg 050801B Egg 050801B Egg 050801B Egg 050801B Egg 050801B Egg 050801B Egg 050901A Egg 050901A Egg 050901A Egg 050901A Egg 050901A Egg 050901A Egg 050901A Egg 050901A Egg 050901A Egg 050901A Egg 050901B Egg 050901B Egg 050901B Egg 050901B Egg 050901B Egg 050901B Egg Sponsor ID Store Name Location 21216 Winn-Dixie Columbus, GA 21216 21217 21217 21218 21218 21219 21219 21221 21221 21222 21222 Winn-Dixie Delchamps I Delchamps I Peggy's Peggy's Piggly Wiggly Piggly Wiggly Kroger's Kroger's Winn-Dixie Winn-Dixie Columbus, GA Mobile, AL Mobile, AL Port St. Lucie, FL F'ort St. Lucie, FL Columbus, GA Columbus, GA Decatur, AL Decatur, AL Decatur, AL Decatur, AL 21223 21223 21224 21224 21225 Haloway Haloway Cub Cub Bi-Lo Decatur, AL Decatur, AL Columbus, GA Columbus, GA Cleveland, TN 21225 21226 21226 Bi-Lo Save-a-Lot Save-a-Lot Cleveland, TN Cleveland, TN Cleveland, TN 21227 21227 Albertson's Albertson's Pensacola, FL Pensacola, FL 21228 21228 Food World Food World Pensacola, FL Pensacola, FL 21229 21229 Fanner's Market Farmer's Market Mobile, AL Mobile, AL 21230 21230 21231 2123 1 Watson's Watson's Food World I1 Food World I1 Cleveland, TN Cleveland, TN Pensacola, FL Pensacola, FL na $ $ . na $ $ Residue Found (ng/g)* PFOS FOSA PFOA NQ - Ah NQ - Ah NQ - AA NQ Ah NQ - Ah NQ - Ah NQ - Ah NQ - Ah NQ - Ah NQ - AA NQ - Ah NQ - Ah NQ - Ah NQ - Ah NQ - AA NQ - Ah NQ - AA NQ - Ah NQ - AA NQ - Ah NQ - Ah NQ - AA NQ - Ah NQ - Ah NQ - AA NQ Ah NQ - Ah NQ - AA NQ - Ah NQ - Ah NQ - Ah NQ - Ah NQ = Not Quantifiable. The residue detected, if any, was below the limit of the study which is 0.5 ng/g. *Value is based on wet weight (5.0 g sample processed as received) ** Values rejected based on unacceptable recoveries. Data has been replaced with either reanalysisor re-extraction values. = Results rejected based on apparent contamination. Data has been replaced with re-extracted values. - = Not analyzed for $ = Sample labels fell off during second shipping and there was ncl way of telling samples apart. Centre Analytical Laboratories, Inc. Page 38 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Table I1 continued: Summary of PFOS, FOSA and PFOA Residues Found in Samples Centre ID 0 106447 0106447 Dup 0106448 0106448 Dup 0 106429 0106429 Dup 0106430 0106430 Dup 0106431 0106431 Dup 0 106432 0106432 Dup 0106433 0106433 Dup 0106434 0106434 Dup 0106435 0106435 Dup 0106436 0106436 Dup 0106437 0106437 Dup 0106438 0106438 Dup 0106439 0106439 Dup 0 106440 0106440 Dup 0106441 0106441 Dup 0 106442 0106442 Dup Data Set No. 050901B Egg 050901B Egg 050901B Egg 050901B Egg 051801AEgg 051801A Egg 051801A Egg 051801A Egg 051801A Egg 051801AEgg 051801A Egg 05 1801A Egg 051801A Egg 051801A Egg 051801B Egg 051801B Egg 051801B Egg 051801B Egg 051801B Egg 051801B Egg 051801B Egg 051801B Egg 051801B Egg 051801B Egg 052101A Egg 052101A Egg 052101A Egg 052101A Egg 052101A Egg 052101AEgg 052101A Egg 052101A Egg na na na na 21213 21213 21214 21214 2 1216 21216 21217 21217 21218 21218 21219 21219 21221 21221 21222 21222 21223 21223 21224 21224 21225 21225 21226 21226 2 1227 21227 21228 21228 $ $ $ $ $ $ $ $ Winn-Dixie Port St. Lucie, FL Winn-Dixie Port St. Lucie, FL Albertson's Port St. Lucie, FL Albertson's Port St. Lucie, FL Winn-Dixie Columbus, GA Winn-Dixie Columbus, GA Delchamps I Mobile, AL Delchamps I Mobile, AL Peggy's Port St. Lucie, FL Peggy's Port St. Lucie, FL Piggly Wiggly Columbus, GA Piggly Wiggly Columbus, GA Kroger's :Decatur, AL Kroger's Decatur, AL Winn-Dixie Decatur, AL Winn-Dixie Decatur, AL Haloway Decatur, AL Haloway Decatur, AL Cub Columbus, GA Cub Columbus, GA B i-Lo Cleveland, TN Bi-Lo Cleveland, TN Save-a-Lot Cleveland,TN Save-a-Lot Cleveland,TN Albertson's Pensacola,FL Albertson's Pensacola,FL Food World Pensacola,FL Food World Pensacola,FL Residue Found (ng/g)* PFOS FOSA PFOA NQ - AA NQ - /\A NQ - AA NQ - AA 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. The residue detected, if any, was below the limit of the study which is 0.5 ng/g. *Value is based on wet weight (5.0 g sample processed as received) ** Values rejected based on unacceptable recoveries. Data has been replaced with either reanalysis or re-extractionvalues. Ah = Results rejected based on apparent contamination. Data has been replaced with re-extracted values. - = Not analyzed for $ = Sample labels fell off during second shipping and there was no way of telling samples apart. Centre Analytical Laboratories, Inc. Page 39 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Table I1 continued: Summary of PFOS, FOSA and PFOA Residues Found in Samples Centre ID 0106443 0106443 Dup 0106444 0106444 Dup 0106445 0106445 Dup 0106446 0106446 Dup 0106447 0106447 Dup 0106448 0106448 Dup Data Set No. 052101A Egg 052101A Egg 052101B Egg 052101B Egg 052101B Egg 052101B Egg 052101B Egg 052101B Egg 052101B Egg 052101B Egg 052101B Egg 052101B Egg Sponsor ID Store Name 21229 21229 21230 21230 21231 21231 na na na na na na Farmer's Market Farmer's Market Watson's Watson's Food World I1 Food World I1 $ $ $ $ $ $ Location Mobile, AL Mobile, AL Cleveland, TN Cleveland, TN Pensacola, FL Pensacola, FL $ $ $ $ $ $ Matrix Egg Egg Egg Egg Egg Egg Egg Egg Egg Egg Egg Egg Residue Found (ng/g)* PFOS FOSA PFOA NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ NQ = Not Quantifiable. The residue detected, if any, was below the limit of the study which is 0.5 ng/g. *Value is based on wet weight (5.0 g sample processed as received) ** Values rejected based on unacceptable recoveries. Data has been replaced with either reanalysis or re-extractionvalues. AA = Results rejected based on apparent contamination, Data has been replaced with re-extracted values. - = Not analyzed for $ = Sample labels fell off during second shipping and there was no way of telling samples apart. Centre Analytical Laboratories, Inc. Page 40 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 12.0 FIGURES Centre Analytical Laboratories, Inc. Page 41 of 151 BACK TO MAIN Figure 1. Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Typical Calibration Curve for PFOS :ompound 1 name: PFOS hefficient of Determination:0.993160 ' Xibration a w e : 7518.25 x + 2045.22 3esponsetype: ExternalStd, Area hrve type: Linear, Origin: Exclude, Weighting: l/x, Axis trans: None Contro &8lytieal L.boratorios, Ina. Centre Analytical Laboratories, Inc. Page 42 of 151 BACK TO MAIN Figure 2. Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Typical Calibration Curve for FOSA - ompound 2 name: FOSA + oefficientof Determination: 0.997449 ahbration cuwe: 29189.8 x 4791.49 esponse type: Extemai Std, Area uwe type: Linear, Origin: Exclude,Weighting: lh,Axis trans: None >.. . .. .... ... ... 010 I ' S!O I ' 16.0 Ti.0 . 26.0 I 2i.O ' 36.0 I 4 0 7 d.0 ' .5.0i.n0g/mL Centre Analytical Laboratories, Inc. Page 43 of 151 BACK TO MAIN Figure 3. Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Typical Calibration Curve for PFOA bmpound 3 name: PFOA Mficiint of Detenination: 0.997082 Ubration CUM: 104281 x + 20236.1 lesponse type: External Std, Area :unmtype: Unear, Origin: Exdude,Weighting: lh,Axis trans: None 523& Response Centre Analytical Laboratories, Inc. Page 44 of 151 BACK TO MAIN Figure 4. Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 ChromatogramRepresenting a 0.5 n g / d mixed calibration standard of PFOS, FOSA and PFOA C101700-7,0.5 ng/mL Standard 021201A-1001 Sm (Mn, 2x3) ' 4 l l - " 13-Feb-2001 14:11:38 LC/MS/MS it6 MRM of 3 Channels ES- 499 > 99 1 .00 2.00 3.00 4.00 5.00 C101700-7,0.5 ng/mL Standard 021201A-1001 Sm (Mn, 2x3) '3. , , ` , ; , , * , , , , , I . , . , I , , , , I , , , . , , , , 1 .00 2.00 3.00 ,,,, ,,,,,,*,, 4.00 5.00 C101700-7,0.5 n@mLStandard 021201A-1001 Sm (Mn, 2x3) 6.00 7.00 Time 13-Feb-2001 14:11:38 LC/MS/MS #6 MRM of 3 Channels ES498 > 78 , , , , , , , , 7T.izm:e 6.00 7.00 13-Feb-2001 14:11:3d LC/MS/MS Y 6 MRM Of 3 Channels ES413 > 369 4.44e5 Area 1 .00 2.00 3.00 4.00 5.00 6.00 7.00 Time Centre Analytical Laboratories, Inc. Page 45 of 151 BACK TO MAIN Figure 5. Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 ChromatogramRepresenting a 2.5 n g / d mixed calibration standard of PFOS, FOSA and PFOA C101700-5,25 ng/mL Standard 021MlA-1003 Sm (Mn, 2x3) 1 3 , * , , 1 . , . , I , , , , I. . , , ) . , ,I , . , - i : , , , I ,I',,,, I,,, , I , 23032 ,'I"' 13-Feb-200114:42:56 L C I I V I ~ S#6 MRM of 3 Channels ES- ,,',,,I ,,., 12605 Time 1.00 2.00 3.00 4.00 5.00 6.00 7.00 1021201A-1003 Sm (Mn, 2x3) 5.85 817351 MRM Of 3 Channels ES498 > 78 4.0705 Area 3.00 4.00 5.00 6.00 7.00 Time C10170M, 2.5 ng/mL Standard 021201A-1003 Sm (Mn, 2x3) ,"','"' I ,,,, I ,,,,,,I , . I ,,,, I.*,,I , , . , , , , )i , , , ~ , * l 375397 13-Feb-2001 14:42:56 LC/Ms/MS #e MRM of 3 Channels ES- , , , , l,;3>z 1.7w 1 .00 2.00 3.00 4.00 5.00 6.00 7.00 Tim Centre Analytical Laboratories, Inc. Page 46 of 15 1 BACK TO MAIN Figure 6. Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Chromatogram Representing a Control Apple Sample (Centre ID: 0009347 Blank A l , Data Set: 020901C Apple) 0009347 Blank A1 12-Feb-2001 18:28:02 lrnJ-- 020901c3009 Sm (Mn, 2x3) LC/MS/MS #6 MRM of 3 Channels ES- 499>99 1.4884 Area 1 .oo 2.00 3.00 4.00 5.00 6.00 7.00 8.0T0ime 2: FOSA 0009347 Blank A1 020901C-3009 Sm (Mn, 2x3) * I 12-Feb-2001 18:28:0 LC/MS/MS # MRM Of 3 Chan~lsES- 3.26 3.633eg6 4*394.61 5.2 5.44 7-27 7.51 7.80 ~.~....~....~....~.,.,~,,.,~,,.,~....~T 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 0009347 Blank A1 020901C-3009Sm (Mn, 2x3) 12-Feb-2001 182802 LC/MWMS X6 MRM of 3 Channels ES- Contra a l ~ i c . 1~aboratorias,Ina. Centre Analytical Laboratories, Inc. Page 47 of 15I BACK TO MAIN Figure 7. Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Chromatogram Representing a Control Apple Sample Fortified with 2.5 ng/g of PFOS, FOSA and PFOA (Centre ID: 0009347 Spk Al, Data Set: 020901C Apple) -: 020901C-3010 Tuttr 0009347 splc ALr 2.5 pob 1: PFOS 0009347 Spk A l , 2 5 ppb OIGl;3010 Srn (Mn, 2x3) 5.21 549311 12-Feb-2001 18:43: MRM of 3 Channels ES499 > 99 2.9085 0009347 Spk Al, 2.5 ppb 020901C-3010 Sm (Mn, 2x3) '3 .oo , , , . I , , , , I , , , , I , . , , I , , , , I 1 2.00 ,,,, I ,,,,I 3.00 , , ) ,1 , 4.00 , ,~, , , , , 5.00 12-Feb-2001 18:43:4C LCNSIMS it6 MRM of 3 Channels ES- 498 w 78 , , " " 6.00 I ,,,, 7.00 Time 8.00 '3,. ,Ill,,,-4.4-7-7 0009347SpkAl,2.5ppb 020901C-3010 Sm (Mn, 2x3) ,,I . , ,.I , . , * , ,,,, I , , I , I , , , I . , , , I , ,,,I , , 12-Feb-2001 18:43:40 LC/MS/MS #6 MRM of 3 Channels E S 413 w 369 ,,I , , ,, I , , ,,I , , Time 1 .00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 Contra U l y f i a a Z Laboratorian. ~nc. Centre Analytical Laboratories, Inc. Page 48 of 151 BACK TO MAIN Figure 8. Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Chromatogram Representing an Apple Sample (Centre ID: 0012927, Sponsor ID: Publix, 18257, Data Set: 020901C Apple) 0012927 020901C-3016 Sm (Mn, 2x3) 12-Feb-2001 20:1258 Lc/MSAmS#6 MRM of 3 Channels ES- 1.6284 Area I~...I....I....,....I..,,I..,.I....rTi~ 1 .oo 2.00 3.00 4.00 5.00 6.00 7.00 8.00 020901C-3016 Sm (Mn, 2x3) V MRM of 3 Channels ES- 1 .00 2.00 3.00 4.00 I""I'"'"'I""I"~~I~'~'I~'~'1 5.00 6.00 7.00 8.00 3 : PFOA 0012927 020901C-3016 Sm (Mn, 2x3) 1 12-Feb-2001 201758 LC/MS/MS #f MRM of 3 Channels ES- . 3.04 I . . ..I....I....,....I.,..I...,I....~~ 1 .oo 2.00 3.00 4.00 5.00 6.00 7.00 8.00 Centre Analytical Laboratories, Inc. Page 49 of 15 1 BACK TO MAIN Figure 9. Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Chromatogram Representing a Control Green Bean Sample (Centre ID: 0009346 Blank A2, Data Set: 020601C Bean) 1S.w: 020601C-5000 `putt: 0009346 D I M f i 020601C-5009 Sm (Mn, 2x3) LC/MS/MS 16 MRM of 3 Channels ES- OOO9346 Blank A2 020601C-5009 Sm (Mn, 2x3) 1 .OO 2.00 0009346 Blank A2 020601C-5009 Sm (Mn,2x3) 3.233.56 07-Feb-2001 08:59:49 1 LC/MS/MS # MRM of 3 Channels ES6 5.94 498 > 78 6.18 1. a 3 6*75 7.27 7 . q q e 2 l....I.~..I....I....I,..,I....I....-~me 3.00 4.00 5.00 6.00 7.00 29717 07-Feb-2001 08:59:4 LC/MS/MS f MRM Of 3 C h a n ~ l eES- 1-. 1.OO 2.00 3.00 4.00 ~ ~ . ~ ~ l . ~ ~ ~ l . . ~ . l ~ . . ~ l , . . . l T.im. .e , , , , , , 5.00 6.00 7.00 Centre Analytical Laboratories, Inc. Page 50 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Figure 10. Chromatogram Representing a Control Green Bean Sample Fortified with 2.5 ng/g of PFOS, FOSA and PFOA (Centre ID: 0009346 Spk A2, Data Set: 020601C Bean) U r n : 020601C-1010 0009346 eplr A 2 8 2.1 oob '4,1: PFOS OOO9346 Spk A2,2.5 ppb 020601C-5010 Sm (Mn, 2x3) , " I . , , , , , . , ) , , . , . , , , , , , , I , , , , I , . , , ) , l;, 1 . 07-Feb-200109:15: LC/MS/MS MRM of 3 Channels ES- ,"I , , , ,I ,,,, 1 .oo 2.00 3.00 4.00 5.00 l j2: FOSA OOO9346 Spk A2,2.5 ppb 020601C-5010 Sm (Mn, 2x3) ..,,I , , , * I , , , * I , , * , I , , , , ) . , , , ,"',,' ~ l ,,, *1i, , "I ,, , ,, , l 6.00 7.00 8.00 07-Feb-200109:15:3 LC/MS/MS # MRM of 3 Channels ES- 498 > 78 ,,,, I ,"',',":"~ Time 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 OOO9346 Spk A2,2.5 ppb 020601G5010 Sm (Mn, 2x3) l&j 606443 , , , , I , , , .) , , , , I , , , , , , , , , ) , , , , ) , , , , I ) , , ,I , , , , 1.00 2.00 3.00 4.00 5.00 07-Feb-200109:15: LC/Ms/MS MRM of 3 Channels ES- 3.3986 Area ,,,, ) ,,,,I,,,II ,,,, 6.00 7.00 8.00 Centre Analytical Laboratories, Inc. Page 51 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Figure 11. Chromatogram Representing a Green Bean Sample (Centre ID: 0012910, Sponsor ID: Bruno's, 18341, Data Set: 020601C Bean) -8 020601C-5020 Taxtr 0012910 0010010 020601C-5020Sm (Mn, 2x3) 07-Feb-200111:53:24 LC/Ms/MS#6 MRM of 3 Channels ES- 499 > 99 6.5864 Area l 1 .00 2.00 3.00 4l.00 5.00 6.00 7.00 zTime 001 291 0 020601C-5020Sm (Mn, 2x3) 100- %- 0.64 0 I I I #*.. ,?. , . , 2 r . , 1 1 I ,T. -n I I II 1 .00 2.00 3.00 4.00 5.00 b 07-Feb-200111:53:24 LCIMSNS #6 MRM of 3 Channels ES- 5.97 498 > 78 6.00 7.00 Centre Analytical Laboratories, Inc. Page 52 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Figure 12. Chromatogram Representing a Control Pork Muscle Sample (Centre ID: 0009348 Blank A l , Data Set: 011501A Pork) -8 0115OlA-1009 Text: 0009348 BluJL Al 0009348 Blank A1 011501A-1009 Sm (Mn, 2x3) . , . , I , . , ,~ 163an-2001 01:48:47 ,i\dn-ii! LC/MS/MS#6 MRM of 3 Channels ES- ~,,,, , 1209 1.oo 2.00 3.00 4.00 5.00 6.00 7.00 Time 0oO9348 Blank A1 1Wan-2001 01:48:47 LC/MS/MS #6 '3,. lee 011501A-1009Sm(Mn,2x3) , , , , , .1,.0, ,0, , , I , , ,2,.00 , , I , , ,3,.00 4.00 MRM of 3 Channels ES498 > 78 Area 5.~00~ . ~ ~ 6~.00. . ~ . l 7..0. 0~ . l ~ . ~T.8.im2Iee3, . , , I . , , , 0009348 Blank A1 011501A-1009 Sm (Mn, 2x3) 16Jan-2001 01:48:47 LC/MS/MS #6 MRM of 3 Channels ES413 > 369 1.3385 Area 1.oo 2.00 3.00 4.00 5.00 6.00 7.00 Time Centre Analytical Laboratories, Inc. Page 53 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Figure 13. Chromatogram Representing a Control Pork Muscle Sample Fortified with 2.5 ng/g of PFOS, FOSA and PFOA (Centre ID: 0009348 Spk.Al, Data Set: 011501A Pork) m: 01150l&-1010 Tut: 0009348 splr A l n 2.5 opb !j,, 0009348SpkAl, 2.5 ppb 011501A-1010 Sm (Mn, 2x3) , . I ,,, , I . , ,, I " ,, ,,,., * , ,, Il,. 'r ,,I,,,,I 16Jan-2001 02:04:26 LC/MS/MS #6 MRM of 3 Channels ES499 > 99 , , I , ~ ,I ,, , , , I , T , , I , , , , Time 1 .OO 2.00 3.00 4.00 5.00 6.00 7.00 8.00 2: FOSA 0009348 Spk Al, 2.5 ppb I '"1 5.81 1494541 16Jan-2001 02:04:2 LC/MS/MS # MRM of 3 Channels ES498 > 78 8.1585 Area " ' 1 . b ' 2.b ' 3 : PFOA 0009348 Spk A l , 2 5 ppb 01119501, A-101.0. ,S,m (Mn, ,,I 2x3) ,, .. I . , , , , . , . . I I . . I - .1 Time 3.00 ' 4.00 5.00 6.00 7.00 8.00 , , , , I , , , , I ,, * ,, : ,a : , 777851 16Jan-2001 02:04:26 LC/MS/MS #6 .MRM of 3 Channels ES- , I , , ,.I ,,, , I , , , I , , , . 413>369 5.2586 Area Time 1 .oo 2.00 3.00 4.00 5.00 6.00 7.00 8.00 Centre Analytical Laboratories,Inc. Page 54 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Figure 14. Chromatogram Represei~tinga Pork Muscle Sample (Centre ID: 0012795, Sponsor ID: Winn-Dixie, 18227, Data Set: 011501A Pork) Bl-: 01150lA-1016 T o x t r 0011795 0012795 011501A-1016 Sm (Mn, 2x3) 1'7 16Jan-2001 03:39:0 LCNSIMS # L 0012795 011501A-1016 Sm (Mn, 2x3) 1Wan-2001 03:39:09 LC/AIISfMS #6 MRM of 3 Channels ES- 498 > 78 5l e; 1 9 , . , , 1 ~ , , 1, , , , I , . . , I , , , , I . , , . I 1 .00 2.00 3.00 3 : PFOA 0012795 4.00 5.00 . 0111350,1,A,-10I 1, ,6,S,m (M* n, ,I . 2. x, 3), I , , , , I , , , , I ~, , , I , , , , I , , , 5*OmL 15654 6.00 7.00 Time 16Jan-2001 03:39:0! LC/MS/IIIIS It( MRM of 3 Chan413n>:els ES. 1Joe! .oo , - . . . I .I I r l I . . . . I . . . . I . . . . Time 1 2.00 3.00 4.00 5.00 6.00 7.00 Centre Analytical Laboratories,Inc. Page 55 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Figure 15. Chromatogram Representing a Control Milk Sample (Centre ID: 0009349 Blank A l , Data Set: 020701A Milk) 1: PFOS OOO9349 Blank A1 I y i l A - 3 0 0 9 Sin (Mn, 2x3) 5.04 99741 09-Feb-20012346:1 LC/MS/MS # MRM of 3 Channels ES- 499 > 99 4.6384 Area 2: FOSA 020701A-3009 Srn (Mn, 2x3) OSFeb-2001 23:46:1 LC/MS/MS #I i MRM of 3 Channels ES. 5.89 498 > 71 A 1sa: 6.59 7.15 1 .00 2.00 3.00 4.00 5.00 6.00 7.00 Time 'rnn OOO9349BlankA1 020701A-3009Sin (Mn,2x3) 09-Feb-2001 23:46:11 LCIMSNS #6 MRM of 3 Channels ES- 413 > 369 1.41e5 Area 1 .oo 2.00 3.00 4.00 5.00 6.00 7.00 Time Centre Analytical Laboratories, Inc. Page 56 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Figure 16. Chromatogram Representing a Control Milk Sample Fortified with 2.5 ng/g of PFOS, FOSA and PFOA (Centre ID: 0009349 Spk A l , Data Set: 020701A Milk) '40009349SpkAl, 2.5 ppb 020701A-3010 Sm (Mn, 2x3) , . , . I , , :, , E) , ' ., ,2, ' 7~ '1I,, ,~, , l 10-Feb-2001 00:01:51 LC/Ms/MSM MRM of 3 Channels ES499 > 99 Time 1 .00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 020701A-3010 Sm (Mn, 2x3) '3, . , , I * , . , I , , , ' ) , , , .) , ~ * ), . , , , ) , , . , I , , , , I , , , , ) , 1 .00 2.00 3.00 4.00 5.00 MRM of 3 Channels ES- . : ;498 78 ,,, ),,, , ) ,,,, 6.00 7.00 8.00 7;;L,,,l.'" 0009349SpkAl,2.5 ppb 020701A-3010 Sm (Mn, 2x3) 'Cj , , , , I , , , .) , , . , I , * , , I , . , , ) , , ,,),, ,,I,,, 10-Feb-2001 00:01:51 LC/MWS #6 MRM of 3 Channels ES- Time 1 .OO 2.00 3.00 4.00 5.00 6.00 7.00 8.00 Centre Analytical Laboratories,Inc. Page 57 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Figure 17. Chromatogram Representing a Milk Sample (Centre ID: 0012871, Sponsor ID: Winn-Dixie, 18202, Data Set: 020701A Milk) -8 02Q70l.A-3014 T u t r 0012871 1: PFOS 0012871 101;A3-024 Sm (Mn, 2x3) I 7 2 : FOSA 0012871 IO;!lA-3024 Sm (Mn. 2x3) 10-Feb-2001 03:42: MRM of 3 Channels ES- 5.05 499 > 99 10-Feb-2001 03:42:3 I LC/MS/MS It Y MRM of 3 Channels ES- A5.84 498 > 78 988 3 : PFOA 0012871 020701A-3024 Srn (Mn, 2x3) 1 10-Feb-2001 03:42:38 1.1885 Area I ~ ~ . . l . ~ ~ ~ l ~ . . ~ I . . . . I . . .T,imIe. . . . 1 .00 2.00 3.00 4.00 5.00 6.00 7.00 Cuitro Analytical trboratorioa, m. Centre Analytical Laboratories,Inc. Page 58 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Figure 18. Chromatogram Representing a Control Chicken Egg Sample (Centre ID: 0009342 Blank A3, Data Sets: 050901A Egg for PFOS, 012401A Egg for FOSA, and 052101A Egg for PFOA) N m : 0509OLA-3010 T e x t : 0009342 Blank A3 1: PFOS 0009342 Blank A3 050901A-3010 Sm (Mn,2x3) 1 3 , ,( , , ( , , , ( . , , , 1 1 , , , , , ~, _I, 09-May-2001 19:16:31 LC/MS/MS #6 MRM of 3 Channels ES- , ,: :( r, ,$ ; Time 1 .oo 2.00 3.00 4.00 5.00 6.00 7.00 8.00 2: FOSA 0009342 Blank A3 012401A-5009 Sm (Mn, 2x3) 244an-2001 19:27:4 MRM of 3 Channels ES498 > 78 1: PFOA 0009342 Blank A3 052101A-3010 Sm (Mn, 2x3) 21-May-2001 17:37:0 LC/MS/MS # MRM of 3 Channels ES- Area % 2.00 3.00 r l I I j , ,I 4.00 5.00 l l l ~ l l l , I I l , . , , , , , , , , , Time 6.00 7.00 Centre Analytical Laboratories, Inc. Page 59 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Figure 19. Chromatogram Representing a Control Chicken Egg Sample Fortified with 2.5 ng/g of PFOS, FOSA and PFOA (Centre ID: 0009342 Spk;A3, Data Sets: 050901A Egg for PFOS, 012401A Egg for FOSA, and 052101A Egg for PFOA) 0009342 Spk A3,2.5 ppb 244an-2001 19:43:27 LCIMSIMS #6 :I,MRM of3ChannelsES> 78 1 .ow Area . . . . . . . . . . , ~ ~ 1 , . , I , , , ~, , , I , , , I , ~, , , I . , . I , 1 , I0 . , I . . , , ~, Time ~ .oo 1 2.00 3.00 4.00 5.00 6.00 7.00 I Centre Analytical Laboratories, Inc. Page 60 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Figure 20. Chromatogram Representing a Chicken Egg Sample (Centre ID: 0012861, Sponsor ID: Save, 18167 and 21226, Data Sets: 050901A Egg for PFOS, 012401A Egg for FOSA, and 052101A Egg for PFOA) N m : 0509OlA-3017 Text: 0106440 1: PFOS 0106440 ,_-;-.l.-I.(z; 050901A-3017Srn (Mn,2x3) 1 3 , I , , , , / ~ , , ,, , I , ~ , , 1 , , 09-May-2001 21:06:31 LC/MS/MS #E MRM of 3 Channels ES- 1.oo 2.00 2: FOSA 0012861 - - .- - -. )012401A-5024Sm (Mn, 2x3) 1OqI 3.00 4.00 I'*"''(' 5.00 ' ~ ~ ' l ' ~ ' ' l ~ ~ ~ ~ l ~ , / , , I ~ I I , 6.00 7.00 8.00 .I 243an-2001 23:24:1 V MRM of 3 Channels ES- 1: PFOA 10106440 052101A-3017 Srn (Mn, 2x3) lo? 21-May-2001 19:26:457 LC/MS/MS #7 MRM of 3 Channels ES- 413 > 369 3.14e4 Area - I " ' I'''~I"''"''l~'~ ' ' ' ' I " ' Time I 1.00' " 2.00 ' 3.00' ' I 4.00 5.00 6.00 I " 7.00 Centre Analytical Laboratories, Inc. Page 61 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Figure 21. Chromatogram Representing a Control Bread Sample (Centre ID: 0009343 Blank Al, Data Set: 020901A Bread) -1 0209OlA-1009 Toxtr 0009343 Blank Al 0009343 Blank A1 020901A-1009 Sm (Mn, 2x3) 11-Feb-2001 03:36:14 LC/MS/MS #6 MRM of 3 Channels ES- 499 > 99 1 .00 2.00 3.00 4.00 5.00 6.00 7.00 Time 0009343 Blank A1 020901A-1009 Sm (Mn, 2x3) 11-Feb-2001 03:36:14 LC/MrnS it6 MRM of 3 Channels ES- 1 .00 2.00 3.00 5.28 4.00 , . . . . I . . s . I q . q ~ I . . . . I ~ . . . I . , . . I , . . T. ime 5.00 6.00 7.00 Contro Analytical Laboratories, Tpo. Centre Analytical Laboratories, Inc. Page 62 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Figure 22. Chromatogram Representing a Control Bread Sample Fortified with 2.5 nglg of PFOS, FOSA and PFOA (Centre ID: 0009343 Spk A l , Data Set: 020901A Bread) '30009343SpkAl, 2 5 ppb 020901A-1010 Sm (Mn, 2x3) , , , . I, , , , I , , , , I , , , , I, . , , I , , , . I , , , , I , , , , I , , , , 11-Feb-2001 03:52:01 LC/MS/MS #6 MRM of 3 Channels ES- 499 > 99 1.oo 2.00 3.00 4.00 5.00 '9,IO09343SpkAl, 2.5 ppb D20901A-1010Sm (Mn,2x3) , , , , , , , , I , , , , I , , , , I , , , , I ,,,,I , , ,,,.,, ,,I ,,,, l , 6.00 7.00 11-Feb-2001 03:52:01 LC/MS/MS 16 MRM of 3 Channels ES- 490 > 70 ~ ~ , , , , Il , , , ,aI , , , . 1.2386 Area ' 1.oo 2.00 3.00 4.00 5.00 6.00 7.00 Time 0009343 Spk Al, 2.5 ppb 020901A-1010 Sm (Mn,2x3) 11-Feb-2001 03:52:01 LC/MS/MS #6 MRM of 3 Channels ES- , , , , I , , , , 1 , , ~ ,~ , 1, ) , , , , I , , , , I , , , , I , , , , 1.OO 2.00 3.00 4.00 g:,,l , , , , ,-,,, ~ 5.00 6.00 7.00 Time Centre Analytical Laboratories, Inc. Page 63 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Figure 23. Chromatogram Representing a Bread Sample (Centre ID: 0012887, Sponsor ID: Albertson's, 18244, Data Set: 020901A Bread) -8 0209OlA-1020 T u t r 0012887 ll j =; : PFOS lo12887 320901A-1020 Sm (Mn, 2x3) , , , , I , . . , ) , ' , ' I . , , , I , , , , I , , , ,~ ~ , , ",' , '), 11-~~tb-200106:29:4 LC/MS/MS #C MRM of 3 Channels ES499 > 9s 1 .OO 2.00 3.00 4.00 5.00 6.00 7.00 Time : F- DSA 012887 11-Feb-2001 06:29:43 120901A-1020Sm (Mn, 2x3) 498 > 78 1.1883 1 .oo 2.00 3.00 4.00 I . . . . I . . . ~ I . . . . I . ~ . . l . , , . I . , . . , . . .T.ime 5.00 6.00 7.00 3 : PFOA 0012887 020901A-1020 Sm (Mn, 2x3) 1 ii-~~tb-200106:29:4: LC/Ms/MS #t MRM of 3 Channels ES- Area 1.00 2.00 3.00 4.00 I ~ ~ . . I . ~ ~ ~ I . ~ . . I ~ . . . I . , . , I . . , , I T. i.m. e. 5.00 6.00 7.00 Centre Analytical Laboratories, Inc. Page 64 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 13.0 APPENDICES Centre Analytical Laboratories,Inc. Page 65 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 APPENDIX A Study Protocol OOP-023-057 (Centre Study No. 023-057), Amendments and Deviation Centre Analytical Laboratories, Inc. Page 66 of 151 . . - BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 .._.... .-.- . .- . . . ... - . . .. . -. . . . . .- . .- -.... Centre Protocol No. OOP-023-057 STUDY PROTOCOL STUDY TITLE .. SPONSOR 3M Environmentd Technologyand Safety Services . Building 2-3E-09 . PO BOX33331 St. Paul, MN 55133-3331 DATA REQUIREMENTS EPA TSCA Good LaboratoryPractice Standards 40 CFR 792 TESTING LABORATORY Centre Analytical Laboratories, Inc. (Centre) 3048 Research Drive State College, FA 16801 Phone 814-231-8032 PROTOCOL IDENTIFI(:ATION NUMBER OOP-023-057 .-- Centre AnalyticalLaboratories,Inc. _ _ -.- _ _ _ ._ - Centre AnalyticalLaboratories,Inc. Page 1 of 81 Page 67 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Centre Protocol No. OOP-023-057 Title: E m k c T I O N OF PFOS, FOSA A N D PFOA FROM VARIOUS FOOD MATRICES FOR ANALYSIS USING HPLC ELECTROSPRAYM4SS SPECTROMETRY TABLE OF CONTENTS Page. STUDYTrrLE ................................................................................................................... 1 1. PURPOSE............................................. .................................................................. 3 2. . TESTMATERM3 ................................................................................................ 3 3. SPONSOR............................................. .................................................................. 5 4. TESTING F A C I L ~ YAND PERFORMBGG LABORATORY ............................. 5 5. PROPOSEDEXPERIMENTALT . I h 4 E - m................................................... 5 6. . TEST SYSTEM....................................................................................................... 5 7. JUSTIFICATIONFOR THE SELECT'I(XIOFTHETESTSYSTEM.................6 8. SAMPLE PROCESSING, STORAGEAND EX3'4TIFICATION........................ 6 9. ANALYTICALMETHOD................................................... 1................................ 6 10. - EXPERIMENTALDESIGN................................................................................... 7 11. - STATISTICALMETHODS TO BE USED .............1............................................. 7 12. PROTOCOL AMENDMENTS AND DEV~ATIONS....._.:.:................................. 7 13. RECORDSAND DATA REPORTING ................................................................. 8 14. QUALITY ASSURANCE ...................................................................................... 8 15. DATA AND REPORT.........................................................................................8 .. 16. ARCHIVE STATEMENT..................... ................................ :.............................. 10 17. REFERENCES.................................. .................................... 1..................:............10 18. PROTOCOLAPPROVAL.................................................................................... 11 APPENDIX: ANALYTICAL METHOD...................................... :................................... 12 Centre AnalyticalLaboratories,Inc. Centre Analytical Laboratories, Inc. -.- ~ - Page 2 of 81 Page 68 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-O23:057 . .. . Centre Protocol No. OOP423-057 1. PURPOSE The purpose of this study is to analyze PFOS, FOSA and PFOA in various food matrices using the analytical method entitled "Method of Analysis for the Determination of Fluorochemicals in Green-Beans, Apples, Pork Muscle, Cow Milk, Chicken Muscle, Chicken Eggs, Bread, Hotdogs, and Catfish by LC/MS/MS". A copy of the method is ,appendedto this protocol (Appendix). The Quality Assurance Unit of Centre 14ndytkalLaboratories, Inc. will audit the study for compliance with EPA TSCA Good Labontory Practice standards 40 CFR 792 (1). 2. TEST MATERIALS The followinganalyticalstandardswill be used: - Test Materiai Lot or TCR Numbe-r Purity (%) PFOS TCR00017-46 - 97.9 PFOA 332. - TBD FOSA L-15709 - 95.1 Expiration Date 08/31/01 07/06/01 02115/01 -Note: G.& characterization,solubility and stability data for PFOA and FOSA are unavailable at this time. However, available characterization data for the test materials will be supplied t6 the testing facility (Centre) by the sponsor (3M). Therefore, do not correct for percent purity when preparing standard solutions for these two fluorochemicals. _ _ . . - _ 1 Centre Analytical Laboritones. Inc. Centre Analytical Laboratories, Inc. Page 3 of 81 Page 69 of 151 I - BACK TO MAIN Centre Study No.: 023-057 Centre Protocol NO: OOP-023-057 ~ .__.. -.-- . -...- .... Centre Protocol No. oOP-023-057 Chemical names and structures of the compounds are presented below. PFOS Chemical Name: Molecular Weight: CAS Number: 0 I-Octanesulfonic acid, 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8heptadecafluoro-, potassium salt. 499 (CgFd03') 2795-39-3 - Note: The neutral molecule and standard form which PFOS (anion) is derived from is perfluorooctanesulfonate potassium salt [CsFi7SO&], molecular weight 538. PFOA Chemical Name: Molecular Weight: CAS Number: Octanoic acid, penkdecafluoro-, ammonium salt 413 3825-26-1 (FC-143) - Note: The neutral molecule and standard.form which PFOA (anion) is derived from is perfluoroctanoate ammonium salt [C7FisCOONHi], molecular weight 431. FOSA Chemical Name: Molecular Weight: CAS Number: I-Octanesulfonamide, 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8heptadecafluoro- - 499 754-9 1-6 A record of test and reference substance receipt, storage conditions, and a record of use will be maintained at Centre Analytical Laboratories, Inc. Chain-ofcustody forms and shipping documents for tracking of the test substances will be included as part of the raw data packag. Centre Analytical Laboratories, Inc. Page 4 of 81 Centre Analytical Laboratories, Inc. Page 70 of 151 . . . . BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 .. . . .. . ... - ..... - _.-. - Centre Protocol No. OOP-023-057 I All standardshest substances and any prepared solutions must be identified with a unique label or number on the container or cross-referenced to the'container. HAZARD INFORMATION A current MSDS for the chemical(s) used in this study will be maintained ai the I - testing facility. 3. SPONSOR 3M EnGronmental Technology and Safety Services Building 2-3E-09 PO Box 33331 St. Paul, MN 55133-3331 STUDY DIRECTOR: Susan A. Beach, 3M TELEPHONE:651-778-7452 . FAX NUMBER: 651-778-6176 4. TESTING FACILITY AND PERFORPdING LABORATORY Centre Analytical Laboratories, Inc. (Centre) 3048 Research Drive State College, PA 16801 PRINCIPAL INVESTIGATOR: Karen Smith, CentreTELEPHONE: 814-231-5032 FAX NUMBER 814-231-1580 5. PROPOSED EXPERIMENTAL TIME-FRAME Experimental Start Date Experimental Termination Date Report Issued January 8,2001 February 8,2001 March 8,2001 6. TEST SYSTEM Food samples obtained by Centre Analytical Laboratories, Inc. and used in the validation study (Centre Study NO. 023-043) will be used the control samples. Samples will be collected from local grocery StOreS from six test cities outlined in Centre Analytical Laboratories, Inc. Page 5 of 81 ~.. ...--.... .- - .. Centre Analytical Laboratories, Inc. . . .. . . _._~ _ . .. . - . . -. . ._.... . . . Page 71 of 151 -- - BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Centre Protocol No. OOP-023-057 3M Quality Assurance Project Plan for Empirical ,Human Exposure Assessment Multi-City Study Sampling Task. All food samples obtained from grocery stores in the six-test cities by the sponsor will be shipped to Centre Analytical Laboratories, Inc. for analysis. 7. JUSTIFICATIONFOR THE SELECTION OF-THETEST SYSTEM Representative samples such as, honey, vegetable (green beans, carrot, radish, lettuce, potato), fruit (Apple), livestock (pork muscle, cow milk), poultry (chicken muscle, egg), fish and processed foods (bread, hot dogs), will be collected by the sponsor and shipped to Cmtre for traCe-k!Vel analysis Of fluorochemical residues. The cities chosen as the collection sites either have major fluorochemicd production facilities, major fluoroche+cal users, or no major fluorochemical production or usage facilities. The matrices were chosen to determine if fluorochemicals are being transferred froni the environment to food sources. 8. SAMPLE PROCESSING, STORAGE AND 1.DENTIFICATION All food samples will be processed accorlding to the method entitled, "Method of Analysis for the Determination of Fluorochemicals In Green Beans, Apples, Pork Muscle, COWMilk, Chicken Muscle, Chicken Eggs, Bread, Hotdogs, and Catfish by LC/MS/MS." Upon receipt at Centre, the fwd samples will -be assigned unique sample identification numbers, which will be used for tracking and identification of the samples. The samples will be stored in a temperature monitored freezer, maintained at < -10" C, except when removed for amlysis as described in the method. The samples will be kept isolated from the test substance during storage. Sample receipt and storage location and conditions -during the study must be documented. All samples and any resu1::ing sample extracts must be identified with a unique label or sample number. Such identification will be either. on the container or cross-referenced to the container. .. 9. ANALYTICAL METHOD The samples will be analyzed according to the analytical method entitled "Method of Analysis for the Determination of Fluorochemicals in Green Beans, Apples, Pork Muscle, Cow Milk, Chicken Muscle, Chicken Eggs, Bread, Hotdogs, and Catfish by LC/MS/MS" (see Appendix). Centre Analytical Laboratories. Inc. ..-- . ._.... Centre Analytical Laboratories, Inc. .. -. . . - .. . . . - .. .... . Page 6 of 81 Page 72 of 15 1 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Centre Protocol No. OOP-023-057 10. EXPERIMENTAL DESIGN I For each matrix, a set of samples will be extracted in duplicate and analyzed. Each set of samples analyzed will Contain at least one matrix control and two matrix control samples fortified at known Concentrations and camed through the extraction procedure to verify recovery. Metho& to control bias will includl: assay of untreated control samples, fortification of untreated control smples to obtain recovery data, and replicate analysis of fortified samples to provide an indication of reproducibility. 11. STATISTICAL METHODS TO BE USED Standard (hear, l/x weighted) curves used to determine the concentration of the compound being analyzed will be constructed from a minimum of six concentrations. Quantification of residues will only occur within the range of the - standard curve (residues in control samples may be calculated up to 20% of. the LOQ [2O% of LOQ = 0.1 ppbl by extrapolation of the calibration curve, in order to correct fortification recoveries). The correlation coefficient (r) for calibration curves generated must be greater than or equal to 0.9925 (320.985). Overall averages, standard deviations and relative standard deviations of fortification recoveries will be calculatedr - Other than these descriptive statistics, no other statistical methods will be used during the course of this study. - 12. PROTOCOL AMENDMENTS AND I~EVIATIONS. Any deviations from the protocol or from the analytical method as provided will be documented and reported promptly to the Study Director. Planned changes to this protocol or to the analytical method *Nil1be made in writing as an amendment or modification and approved by the Principal Investigator and the Study Director. Any amendments will be appended to this protocol and included in the final report. 1. Protocol amendments: Planned changes to the approved protocol shall be documented by amendments that c1e:uly describe the change, justification for the change, and impact on the study. Amendments will be signed and dated by the Principal Investigator and Study Director. Copies of amendments will be sent to the quality assurance Unit. 2. Protocol deviations: Protocol deviations, which are one time and unplanned . deviations from the protocol shall be documented in the study'records, noting the nature of the deviation, potentid effect or impact on the study, and corrective action if required. Protocol deviations are signed by the Principal Investigator and Study Director and reviewed by QAU. . Centre Analytical Laboratories, Inc. I Page 7'of81 .. Centre AnalyticalLaboratories,Inc. Page 73 of 151 .. .- .- BACK TO MAIN Centre'StudyNo.: 023-057 Centre Protocol No: OOP-023-057 . ... .. . . . . .. - Centre Protocol No. OOP-023-057 13. RECORDS AND DATA REPORTING Records to be maintained include, but are not limited to the following (as appropriate): 1.Sample trackin-g sheet($ 2. Sample receipt records, storage history, and chains of custody 3. History and preparation of standards (stock, fortification, calibration) 4. Description of any modifications to h e method 5 . Instrument run sheets, bench-sheets or logs 6 . Analytical data tables - 7. All chromatographic and instrumental conditions 8. Sample extraction and analysis dates 9. A complete listing of study personnel, sisatures and initials 10. Chronological presentation of all study correspondence 11. Any other data necessary for the reconstruction of the study All chromatopms will contain the following: a. Sample identification, date, arrow or other indication of the area of interest, and injection number corresponding to the run. b. Additionally, fortifications will include the fortification level of each analyte. c. Analytical standard chromatograms will additionally include the concentration (e.g., pg/ml, n g / d - , ppb, ppt. etc.). Each data set will contain information on temperatures, flow rates, column parameters, gases, instrument parametea;, and instrument type, etc. if any of these differ from the method. The Centre study number will also be recorded on the first chromatogram of each daily ana1yiic:al run. 14.. QUALITY ASSURANCE Centre QA Unit will review the protocol and audit the study conduct and the study documentation (including raw data and final report) to ascertain that all QNGLP procedures are adhered to. Ckntre QA Unit will inspect the study at intervals adequate to assure compliance to GI-F"S, and will report the findings of the audits to the Principal Investigator, Centre Management, and the Study Director. 15. DATA AND REPORT 1. All raw data and the original signed protocol will be maintained i; the study file. This data includes the protocol amendments, protocol deviations, laboratory notebooks, analytical sthdard solution preparation, sample chain of custody sheets, sample work sheets, chromatograms, calibration curves, and any other appropriate data generated. Raw data not used will be stored in the study file. The reason for exclusion will be documented. Centre Analytical Laboratories, Inc. Centre Analytical Laboratories, Inc. Page 8 of 81 Page 74 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 ... . ... . . - .- - .. .. . . .. .- - Centre Protocol No. oOP-023-057 2. A final report will be issued by Clintre, approved and signed by the Study Director and sufficient for submission to EPA. The report contents should include, but are not limited to: 1. Objectives and procedures stated.in the protocol 2. Analytical and statistical methods used 3. Test materials identified by name, lot, purity and other characteristics 4. Name of testing facility and study initiation and completion dates 5. Statement prepared andsigned by the quality assurance unit 6 . Tables containing all applicable data 7. All chromatographic and instrumental conditions 8. A complete listing of Centre study personnel . Centre will send a copy of the draft report to the Study Director, who will return the report with comments. Centre Analytical Will make revisions and finalize the - report with the approval of the Study D:irector. The Centre QA unit will conduct an audit of the final draft report and data files to assure accuracy and GLP compliance. The final report and copies of the raw data must be approved by the - Study Director prior to signature. A statement of inspections and a statement of compliance will be included in the final :report. Any corrections or additions to the final report shall be in the form of dn ame&nent by the Principal Investigator. The amendment shall clearly identify the part of the final report that.is -being corrected and the justification for correction. The amendment shall tle signed -and dated by the Principal Investigator and the Study Director. REPORT DISTRIBUTION: Original: Study Director Copy: Centre Archives I Centre Analytical Laboratories, ~ n c . Centre Analytical Laboratories, Inc. - __ . Page 9 of 81 Page 75 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol,No: OOP-023-057 Centre Protocol No. 0013-023-057 I 16. ARCHIVE STATEMENT Study records to be maintained: Records to be maintained for the study include all raw data, observations recorded during the conduct of the study, documentation, chromatograms, specimens, and study related correspondence. This includes a '- description of equipment used during the conduct of the study. All characterization data and any shipping records shall be retained. Document archives: Upon completion of the study, the study records, protocol and - amendments, and the final report and amendments shall be retained in the 3M document archives. If it is necessary to substitute a copy for an original record, it will be certified as an exact copy. Centre -will retain facility-related originals and a copy of the final report and raw data package. 17. REFERENCES 1. U.S. Environmental Protection Agency, Toxic Substances Control Act (TSCA). Good Laboratory Practice Standards. Final Rule, 40 CFR Part 792. Centre Analytical Laboratories. Inc. Centre Analytical Laboratories,Inc. - ._ Page 10 of 81 .. . Page 76 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Centre Protocol No. OOP-023-057 18. PROTOCOL APPROVAL This protocol was audited by the Quality Assurance Unit of Centre Analytical Laboratories; Inc. Quality Assurance Reviewer, Centre I %baa B. Kinne 1 Qudity Assurance Auditor b! /03/0 1 Date Principai Investigator, Centre Facility Management, Centre .. . . Laboratory Manager President 6 /e&+ Study,Director, 3M VL-. Susan A. Beach Sp o n i s o T23M - Dale acon ' Technical Director Centre AnalyticalLabontories, Inc. Centre Analytical Laboratories, Inc. 03-@-6I Date ~,~ .i I Date Page 11 of 81 Page 77 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: 00P-023-057 Centre Protocol No. oOP-023-057. APPENDIX: ANALYTICALMETHOD METHOD OFANALYSIS FOR `rmDETERMINATION OF FLUOROCHEMICALS IN GREEN BEANS, APPLES, PORK MUSCLE, COW MILK; CHICKEN MUSCLE, CHICKEN EGGS, BREAD, HOTDOGS, AND CATFISH BY LC/MS/MS. . . - - ... Centre Analytical Laboratories, Inc. Centre Analytical Laboratories, Inc. Page 12 of 81 Page 78 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 - -. .. Centre Protocol No. OOP-023-OW-- .. .I TITLE Method of Analysis for the Dertmination of Fluorochemicalsin Green Beans, A ~ ~ ] ~ ~ , Pork Muscle, COWMilk, Chicken Muscle, Chickm Eggs, Bread, Hotdoss, and Catfish by LCIMSIMS I .. -AUTHORS . Enaksha Wickremesinhe,Shaozhi Zheng and John H & e q DATE ISSU@ September 13,2000 -SPONSOR . . . jhl Environmend Technologyand Safety Services Building 2-3E-09 -POBOX33331 . ' St. Pad, MN 5512'3-3331 PERFORMINGLABORATORY Centre AnalyticalLaboratories,Inc. (Centre) 3048 Research 1)rive State College. PA 16801 C E m STUDYNUMBER 02340.1 . TOTAL 7U'UiMBER O'FPAGES I " I Centre Analytical Laboratories, Inc. Page 13 of 81 Page 79 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 ....... . . -. . - . . --.- . . . ........... .... 4 ._.. :. .. . . ..- . .- . . --_ .. _ _.. . .. .. Centre Protocol No. 00P-023-Oj7 : '. ... ... . Author, Principal Investigator Centre Analytical Labontories, Inc I\ .. 9-13-00 Shaozhi Zheng Author, Principal Investigator Centre Analytical Laboratories,Inc. Date , . $,h; I._. .. Date .. ' hboratory Manager Centre Analyticai Laboratories,Inc.. .. . . .. . -- R i c h s d k Grauini, PhD. Date President' . Centre Analytical Laboratories,Inc. . - , Hans.cn, PhD. . Date SponsorRepresentative '. 3M Environmental Labontory .. CenrreAmlyrical kbontorics. Inc. Study C 023601 Page 2 of 69 I , . Centre Analytical Inboratoiies. Inc. I ' Page 14 of 81 .. . . . . . . . . . . . . . . . . . .. . . . . ...._ .._._......, .__._:... Centre Analytical Laboratories, Inc. ---_ --- _ _ _ - _ _ - Page 80 of 151 .. . . ............ ... BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 . . . ... . . ............ . . . . . ...... ......... ... -- .. . . . . ... . . Centre Protocol No. OOP-023-05.7. : .. '. TABLE OF C 0 " T S ?rrr;EPAGE .................................................. ................1.............................. ..................... 1 WAG- AppROVAL ............................................................ ............................... 2 TABLEOF c o r n s...._..:............ ~........... ~ ............................................. 3 LIST OFTABLES ........... :.............................................................. :........................ 4 LISTO F R G W ............._.i................................................................_.............. 5 1. SUMMARY ...................._.:..................................... ............................................... 7 2. COMPOUNDS...:.... ....................: ........i ......... ................................. 8 3. .bJvD S ~ P ..W................................................................ ................9 .3 . 1 ; . m w ..........__............."............................................ ............................... g 3.2. STANDARDS?.e................ ................................................ g . . 3.3. E ~ J ~ , W A M........)......~................................ ................................... 10. . 3.4. S O L ~ O ~ ~ . . ............; ................................................................ ........................ 11 . 3.5. mcx PREPARA~ONOF FOR~XATIOX C-U~ON S O L ~ O N....~...11 . 3.j.1. Stock Soludons...................................... ........................................... 11 . 3.5.2. Fortifidon SOlUuOnS.............................................. ......................... 11 . 3.5.3. Cdbntim Stanndards.............................................................................. 12 4. METHOD.............. 1.................................................... .................................. 12 .: . 4.1; OWD DUG^................................................................... .............................. I2 .- . 4.2. S&p~pRo(zssING...; ...................................................... ................................ 13 . 4.3. O ON.......-.... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ...................:..........13 .. 4.3.1. Apple, Green Beans, Bmd, Ho~!DoiLand Pork ........I........... 13. . . 4.3.2. Chicken mdMilk ............................ ....................... ........................... 14 . 4.3;3. E% and atfish........................................ :........................................ 1j 4.3.4. SpE Column PrcpantiOn................................................................... 16 . 43,5. Sil&adon of Pear-Shaped FMcs................:.............................16 . 43.6. Sm&-&arion ofSPE column^ ...................................................... 16 4.4. & ~ ~ ~ ~ B Y L C / M S.../.:.M...._S .............................................. ..................... 16 . .. 4.4.1. h C i / M S System and openti% (hditions filectrospmy).........16 . 4.4.2. h m p l e Tunc File Pmmten ............................... ........................... 18 4.4.3. Cdibntionh c t d u m .............................. :.-........... ..............19 4.4.4. samplehdysis.-........................................................ ........... 19 4.j. pERFO&\WE ................................................................. ...................20. . 4.6. &QVIR~,A)S ANI\LySIs............................................. ............................ 21 . 5. C A L W n O N S ................................. 1............ .........................................21 5.1. C 0 m m - r m b i CokcEEinunON...............2................I............. 21 5.7.. F O R ~ ~REC~OVEORYN...................................................... ....................... 22. 6. s m............................................................................................ ........................ 22 .. . Ccnm Analydal Lbonrorics, IncSNdy 03001 p3gc 3 of 69 ... Centre Analytical Laboratories. Inc. ............... .. ..... . Page 15 of SI ..... ... Centre Analytical Laboratories, Inc. ...... ........ . I Page 81 of 151 BACK TO MAIN I. .. ........ Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 . . . . . -. . . . . . . . - . CentreProtocol No. OiP-023-057 -. .. I LISTOFTAI3LES . . . Table 1: Su-~y ofRecoveries for Et-FOSE-OH, Me-FOSE-OHjFOS, p o u and pK)SA in Whole Apple............................................................... -74 Tiible 2 Summary ofKecoveries for Et-FOSE-OH, Me-FOSE-OH,PFQS, p0,A.A a d PFOSA in Ge:n Beans'........................................................ .......... 25 -Table3: Summary of Recoveriesfor Et-FOSE-OH, Me-FOSE-OH,PFOS, POAA and PFOSA in Chicken E,QUS .........................................:..................... 26 Table 4: Summaryof Recoveries for Et-FOSE-01%Me-FOSE-OH. PFOS, p0,A.Aa d PFOSA in Chicken Muscle............................................................ 27 . Table 5: Summary of.Recoveries for Et-FOSE-011, Me-FOSE-OH, PFOS, ' pOAA and PFOSA in cow Milk .............................................................. .......28 . Table 6: pSuommuaarnydopfFROecSoAveinrieBsrfeoirdE..t.-..F..O:..S...E...-.0...1..1...,..M....e...-.F...O....S...E...-..O....Y....P.FOS,...................2.9 . Table 7: Summary ofRecoveries forEt-FOSE-OH, hfe-FOSE-OH,PFOS, POAA and pF0S.A in Hot DOg.'........................ ........................... ........... 30 Table 8: Summary of RecoveriesforPFOS, and F'FOU in Cmish .................31 . Table 9: SummaryofRecoveriesfor Et-FOSE-011,Me-FOSE-OHJFOS, .. pOAA and PFOSA in Pofi'Muscle ................................................................ 32. -- .. .. -.. - .. ... . Centre Analytical Laboratories,Inc. ' . . . . . . Centre Analytical Laboratories,Inc. .. - . . Page 1 6 o f 8 l Page 82 of 151 BACK TO MAIN .. . Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 ... Centre Protocol No. COP-073-057 I .. - _ _ _ _ C_ _e_n_tr_e_A_n_alytical. Laboratories, Inc. .__-_.- . ' . _-.-... Centre Analytical Laboratories, Inc. , .. .. ... .. __ . Page.17 of 81 ' ..... Page 83 of 151 .. BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 .. Centre Protocol No. OOP-023-057 . ~ ~ I LIST OF FIGURES (:CONTIiNTJED) F = p e 17: Repnsentative Chromatogmm from the Injection of Control Catfish Fodfied at j00ppt each of PFOS, PC)hl, and PFOSA (Set 012$00 Catfish, C e n k ID99120.52 Spk A) ............................. ................................ 65 Figure 18: Representative Chromato-guns of a Control Pork Muscle Sample (Set 172199 Pork, Centre'D 9911783 Pork Blank B) ........................ .......... 66 Figure 19: Repmentauve Chromatogruns from the hjxtion OfControl Pork hiuscle Fonified at 500 ppt each of Et-FOSE-OH. M:-FOSE-OH, PFOS,POAA, . and PFOSA (Set 122199Pork, Ccnm ID9911783 Spk B)................ ..........68 ._. Centre Analytical Laboratories. Inc. Centre Analytical Laboratories, Inc. ..... pap: 6 Of 69 'Page 18 of 81 .. Page 84 of 151 .. . . . BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 .. . . . .. . . . . . . . . .-_ . . . Centre Protocol.No. OOP,O23-057 ___ .. I Centre AnY!yticalLaboratories, Inc. .. Centre Analytical Laboratories,Inc. ---.- - ..... . .. . Page 19 of 81 Page 85 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 . _. . . .......... Centre Protocol No. OOP-023-057. ... .. .. . I ,. 2. EXPERTILIENTAL COMPOUNIIS Molecular structures of Et-FOSE-OH.Me-FOSIXW PFOS, POA.4, and PFOSA are given below. .. .. PFOS molecular weight: 499 (CgF17S037 ' - - ChauicalNanc = P c d u o m n r ~sculfonate ....... -----.----.-- I Centre.A. nalytical Laboratories, Inc. ........ ... ..... ..._---.. . I .. .... (. . Page 20 of 81 Centre Analytical Laboratories, Inc. . ~--. . .- ..- .._.--. .-. . -. .. . . . . . . . -.. . . . . . Page 86 of 151 . - ........ . .. . . "........ _ . - .. , BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 .. - ....... ......... __ ... ..... ._ - Centre Protocol No. OOP-023-057 - ... .___ PFOSA molecular weight: 499 0 3. cHEMIc.QLs SUPPLES 3.i. CIEMICALS . ' . . Chemicti Acetonide (ACN) Carbon .. . . Methmol~eOH) . Silica gel (60-200 mesh) FIon'siI(60-100 mesh) S uperclem'LC-N& Gnde . HPLC 120/400 HPLC Reagent Reagent Reagent Source m)J.T. Baker . Supelco 0J. T. Baker m)J. T.Baker J. T.Ba-k.er Supelco Catalog Nr JT~OI~:~ 57210-U n9093-2. . 3405-0j 3372-08 .5-7205 I-Octanol Ammonium Acetate L-Ascorbic acid Reagent Reagent .Reagent Aldnch Chemical . Si,gna-Aldn'ch - . Si-m-AIdn'ch 111-57-5 ~-7330 L-5960 . Dimetyldichlorosilane Toluene ... Water Reagent Reagent Type1 - m) SuPelco J. T.Baker inhouse 3-3009 J'l-946013 nype I water = e!ectricd resistivity,minie:um of 16.67W c m at 25C. from a Lbconco waterpro workst3tion) 3.2. STANDARDS Standad Et-FOSE-OH Me-FOSE-OH PFOS POAA PFOSX . 1 Gnde - Andytid Analytical . AildytiCal Analytical AndvdCal Source 3M 3M 3M ~ 3M . 3M Centre Analytical Laboratories, Inc. ................ .... .. .. 9 Of69 Page 21 of 81 Page 87 of 151 .. BACK TO MAIN .: , .. .. ........... Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 ............. ......... ........... Centre Protocol NO. OOP:O..2..3.:.0..5.7.. _-... _. 3.3. EQUIPMDTANDSUPPLES EQUIPMENT SOURCE Balance, analvrical Balance. top-loading ' Rotary ev$orator .. Rotary evaporator trap Tissumizei . Sorvall RC 5C plus Centrifuge ' Mett~ ler.-_ _ Mettler . Buck : Buchi . Tekmar Dupont IEC HN SII centrifuge IEC Pm-shaFd flasks . SPE tubes (20 ILLc)at no. NO57177 PPX Supelco Vacuum pump . Buchi Visiprep vacuiim manifold Supelco Wrisi-action s@cr. . Burrell Bench-top centrifuge 50 mLdisposable polypropylene.centlifuge tubes . . Disposable pipei, tes! tubes etc. .. IEC VWR VWR- 125-mLLDPE narrow-mouth bottles Nalgene 2-mZ.~levHPLCvialkit(catn'5181-3400) Standard lab equipment (clgs A pipets and - . - -. H P volumetric flasks,graduated cylinders, etc.) . , L U M S ~ S -and HPLC systems are . . As described in Section 4.4.1. - __ Centre Analytical Laborat+eS, Inc. .. . . . . Centre Analytical Laboratories, Inc. .... .. Page 22 of 81 .. . . ... .. Page 88 of 151 BACK TO MAIN ._. ~ . . .. . Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 ... -. - . .. Centre Protocol No. . O...O.P..-.02.3.-_05_7... . ,. Centre Analytical Laboratories, Inc. .-.- .... . . . .- __ - .- . .. ... . . . ... .. . . , . _-.- Page 89 of 151 . .. . . .: . . . .. BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Centre Protocol No. OOP-023-057 --- I 1 .o 2.5 100 * 0.025 1.0 1.0 100 . 0.01 0.05 10.0 100 0.005 0.025 10.0 ' 100 0.0025 0.01 10.0 - 100 0.00 1 0.005 10.0- 100 0.0005 each of Et-FOSE-OHh,.lc-FOSE-OH.PFOS, POAA,and-PFOSA - - -- - ... ..... 4. METHOD 4.1. FLOWDUGRAM - The flow &;gam of the method is g h I k l O W , followed by a 'detailed description of a c h step. . . .. . 5 g of matrix. Fortify if needed 1 E.xtnct with ACX 3. .. . SPE column clean up :1 . Concenmte to dmost dryness .. 1' !. Adjust final volume 5 LCRrlSh4.S analysis ! '. . Ccnrre h d p i c d ~ . & ~ r Iinc.~SN, dy 3 I I pa& 12 of 69 ' , Centre Analytical LaMratorics, ~ n c . .-. .. . .I. Page 24 of 8 1 Centre Analytical Laboratories, Inc. - -. Page 90 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 Centre Protocol No. COP-023-057 ,_-. .-. . .. Centre Analytical Laboratories, b c r Centre Analytical Laboratories, Inc. ! I.. ' Page 25 of 81 .. Page 91 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 b. C. 'J, Centre Analytical Laboratories,Inca . ..... .. . . ......... Centre Analytical Laboratories, Inc. .. ..... .- . Page 14 of 69 Page 26 of 8 1 ...... Page 92 of 151 BACK TO MAIN - ... .- - .... .. ..... _. .. Centre Study No.: 023-057 .CentreProtocol No: OOP-023-057 . .- . .. . . . . . ~. Centre Protocol No. COP-023-057 . . .. , - 1. ... . .- . . ..-__-. -. ._ .. - ' .. . . Centre Analytical Laboratories,Inc. -. .--- Page 93 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 - - - - . - -. _ _ _ _ Centre Protocol No. COP-023-057 - .. , , I .. . I a. b. C. ' d. : e. f. Centre Analytical Lhomtories, h c . ' .. .._..-... . .. . .-.-- Centre Analytical Laboratories,Inc. . .~... . . . . . -.- ! :P&lhof69 I I .. I 'Page 28 of 81 , .I *.I -. . . ...- .. , Page 94 of 151 - .. BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 .. .. .. I - .. . .,..:- ...-... Centre Protocol No. 00PT023-057 . ....-_....._ ~ HPLC: Hewlett Packard (HP)Series 1100 HP Quat Pump Hp Vacuum De=w s e r HP Autosampier . Hp Column Oven - , Note: A 4 x 10 mm hypermi dropin guard canridge (Keystone, parr ::841017400) is aaachecl on-line after the purge valve and before the sample injector pOn 10 m p my residue contaminants [hat may be in the mobile phase and/or HPLC system. HPLC Cslury: Genesis CS (Jones iChromatopphy),2.1 rnm ColumnTemp.: 35" C . Injection V O ~.: IO PL . MobilePhase (A):? mMAmmonium Acetat in Type I water Mobile Phase (B): Methanol 50 rnm,4p - Time .o - % A 60 ' . -% B 40 0.4 . 60' 40 1.0 - 10 90 7.0 10' . ' 90 75 - : 9.0 - 0 - . . 100 0 100 . 95 135 60 40 60 . 40 . 14.0 60 40 0.i00 0300 0300 0.500 0.400 0.400 0.400 . 0.300 Ions monitored: - Analvte Et-FOSE-OH Me-FOSE-OH PFOS POM . PFOSA - Mode negative negative negative negative negative - Tnnsition Monitored 6ZlO,+J 59' 616 9 59 499 3 99 413 369 - 498 9 78 Approximate Retention.Time 67 .a. I 6.7 5.3 . . 5.1 . 6.0 - . . . . ... . ! Centre Analytical Laboratories. Inc. , t i . . . . . .. .,. . . .. ._ -. I . ~ .. Centre Analytical Laboratories, Inc. .- .. 17 of 69 Page 29 of 81 . . ..__. . ... Page 95 of 151 .. . . -. . . . . BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 - .. .... . - Centre Protocol No. OO.P--023-057. I Arialvte Dwell (sl Et-FOSE-OH . 0.2 Me-FOSE-OH - 0.2 PFOS . 0.2 - POAA . . 0.2. PFOSA 0.1 ' -S7 ource . Capiilary Hexipole 1 - . Awl Hexawle 2 Sourcc Block Temp. Desolvation Temp. . -Collision E n e r p (em 31 Cone 30 31 30 43 76 11' 25 2s 34 - - Set. 2.56 kV . O S V - .- 0.1 v 0.8 V . lo0T . 400T . -Set . 13.0 V 13.0 V . 0.7 V . -2v IV 11.ov . ll.0V 1.0 v . 650V , G s Flows 'ConeG~ . Desolvation .I Pressures GY Cell I Centre h i y t i ~ hdb o n t o r i a k S t u d y nY E5.j-001 . I Set -1 5 F ! - 700 I/hr -Read back 3.Oe-3 mbar Centre Analytical Laboratories,inc. .- . - ._ . .. - .. . .--.. Centre AnalyticalLaboratories,Inc. .I Page 30 of 81 Page 96 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 - - - - - _ - I _ .: Centre Protocol No. OOP-023-057 ~ .. ... . ... Centre Analytical Laboratories, Inc. ._ . . . . . _- Page 97 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 .. '. .. . . ~ .. . , , -. - . .. . . - - .. -- I _ .. . Centre Protocol No. COP-023-057 .. ._. I ' Centre'Analytical 'Laboatories, Inc. ,: Centre Analytical Laboratories, Inc. Page 32 of 81 Page 98 of 151 BACK TO MAIN Centre Study NO.: 023-057 Centre Protocol No: OOP-023-057 -- .- . . ..... . . - . . .- . .. .. Centre Protocol No-..OOP-023-057 .-- 5. CALCULATIONS Equationk . Determine the component residue concentntion using Equation 2. Equation ?: c=-!~Atnalytical Laboratories. h c . -. .- . . .. . . . - - .. . ._. - . .. . . . .. . .... Centre Analytical Laboratories, Inc. . 'Page33of81 ' ' . .. . . BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: 00P-023-057 ._ Centre Protocol No. 0013-023-057 5.2. FORTIFICATRIOENC O ~ Y Y - Equation 3: .. Recovery (%) = - (ndg found (avengen&/:ifound in control)) ' ndgadded XI00 or 6, SAFETY . ._ Centre Analytical Laboratories; Inc. .' . . -. . ... . -. -- Centre Analytical Laboratories, Inc. Page 34 of 81 '- .- --__ _. Page 100 of 151 ..... BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 .................... - - - - - .---- - Centre Protocol No. OOP-023-057 ... .. ... :TABLES . . ... Centre Anal)tical Laboratories. he. .... ..... - .- . Centre Analytical Laboratories, Inc. ........ ........ ......... Page 13 of 69 ...... Page 35 of 81 Page 101 of 151 BACK TO MAIN , ... 6 'I'nhlc 1: S I I I I I I oIfII~tc~coYvcrics for IZITOSE-011, ILlc-FOSII-011,I'FOS, POAA n t d I'FOSA in \Vl~olcApplc E-OH Trlal 2 Me-FOSE-OH ' PFOS . Trial 1 Trlal2 PI TrlJl 1 11111K Trld 2 PF Trlnl 1 I SA - - Trlrl 2 . 111 89 irk ND NQ ND ND 0.432 0.411 0.892 0.840 1.787 1.781 4.514 4.787 9.100 I - 9.024 09 0.134 a2 91 '6 ' 5 41 .. . 1 z. W o\ % '%' 00 c c % 0 N .. .- c VI c Celilre Amlytical I.aliornlorics. Iiic. Sliitly N 023-001 I I I I ! f 1':igc 24 of 63 i I I I I BACK TO MAIN i : I cm en : iE rn i i" |8 BACK TO MAIN F.' Y,. A' El-FOSE-OH Trlal 1 Trlal 2 114100 : hde-F( Trlal E 1W20180 i .! . . . . . . '. . Reegenl Blenk B Malrlx Blank A NO Malrlx Blank'B NO 1.0 ppb Spk D 2.0 ppb'Spk F 10.0 ppb Spk I AVERAQE NO NQ - * NO NO * NU 0.311 83 0.492 08 0.615 81. 0.831 03 1.po 69 1.206 60 3.642 71 4.4OQ 88 &OS0 se - 10.030 8.708 87 - 10' 7G - 16 15 L 15 !' . I I !I .: w c1 mrD W 00 % m c-. . a . Ccnlrc Arinlylical ~.~bnraloricsIn.c. Sliidy It 023-001 .~ * . 8 a . PI Trlrl 1 A Trlal 2 I- & PI Trlal 1 Trlal 2 NQ NO Nil NQ NO NQ NO k.rb EIQ 0.465 03 0.302 78 0.436 0.488 08. 0.380 00 0.466 01 1.1 11 111 0.898 60 1 . 0 ~ 2 , 1.163 115 0.042 04 1.120 2.060 103 1.683 79 2.138 107 1.081 90 1.726 86 2.145 107 4.623 W 4.004 07 5.203 4.807 98 P.038 101 6.410 0.120 01 8107 08 - to.lra 101 0.220 02 - I 11c 100 OB - - - 9 8 0 10.400 .. - ,. .. h g c 26 01-69 I . . :,I /, ! .- .. .' I BACK TO MAIN G , , p 2 0 3. Ei 0 .. I . Et-F( Trlal 1 11f20/99 5-01-1 Trlil2 11/22199 Aeagenl BlankA Roagonl'Blank B Malrlx Blank A ' Malrlx Blank B ND 0.5 ppb Spk A 0.5ppb Spk B 1.O ppb S p k C . 1.0 ppb Spk D 2.0 ppb Spk E 2.0 ppb Spk F 6.0 ppb Spk G . .. , 10.058 AVERAOE 95 101 7 40 Trlal,e-FjSE-OH Trlal2 p9 42 53 60 '66 60 62 --85 ,, -80 1 O( 66 10 ! ~ PFOS P Trld 1 - 11/21 9- - WtL nai ND ND NO NQ D.488 84 0.470 84 0.881 88 0.669 07 1.921 98 1.848 82 4.761 as 4.628 81 - - 10.166 101 9.920 88 95 5 \A' I Trlol 2 94 6 PF Trlal 1 1 1120/88 I 06 10 Trlal 2 I . .. . . . .- , -s00 Ccnlrc Aiialylicnl 1-nbornlorics. Inc. Sliidy N 023-001 I'ngc 27 of G9 BACK TO MAIN ! I ! .. '2 1 ;5 F : ' ' 9 . , . . . 1. -. .- -. . -. - j 'I'JIIIIC 5: Suiiliiinry of Rccoverics for El-~os~-ohIfcI-I,POSE-OIi,I'FOS, POAA rind I'FOSA i t I C e ~ Mv illc Mc-F Trl.1 1 llI9B9 iltl F Trl.1 1 11mm9 I. T r l i l 2 ' llIlU&S !!!YL &L NO .453 NO NQ ,335 ND 8.332 4.417 7.832 3.449 D.480 1.030 0.908 1.785 1.971 5.380 6.314 0.168 .7 g ' 1.552 44. D.604 101 1.570 39 0.632 105 2 1 7 88 0.951 05 1.018 68 0.007 87 1.844 68 1.578 79 1.038 80 1.787 88 4.800 4.810 89 ,4.02e 99 Ro , Ie?? 93 0.388 01 5.782 88 - o . a e 01 8 2 6 2 53 10.880 I . 7 5 93 1O i 1 10 20 8 8 -. .- . . .- .. .. PI Trlil 1 llmloo 8.200 8.152 00 7 . ... ;A Trial 2 lltl8B9 -. .. - ._ . .- 1.423 .. .. 2 0,. . 0o - 0 z 0 Cciilrc Aiialylicnl Lnlmratorics, Iiic. Stiidy N 023-001 . I I . .. I BACK TO MAIN .. I ' ' ' IC ; 5. !. . -2 . IF 0 I .. RosgenlBlankE Malrlx Blank B 0.5 ppb Spk A 0.5 ppb Spk B 1 .O ppb Spk C , 1.O ppb Spk D 2.0 ppb Spk E 2.0 ppb Spk F 5.0 ppb Spk 6 5.0 ppb Spk H .n n I"." _ I.#'_ " L q_,_mI _ I . El-F . Trlsl 1 -1I/ 9I) 32IL (R.l , ND iE-OH Trial 2 11118/99 ND ND' NO 0.356 71 0.305 77 0.058 88 0.047 05 1.728 88 1.700 85 5.188 .4.8n8 - i i . 0 i i 0.708 103 88 -110 87 /'4::g;i 91 12 973 103 9 Me-F Trlal 1 11/16/90 . 07 a10 ;-OH Trlal2 -111 - 22% NO' )o L net NO NO NO 0.4 18 84 0.423 85 0.038 04 1.071 2.010 2.001 4.707 4.814 10.031 107 101 105 08 88 -100 111 00 9 P - . Trlall 111 rn 3t!L 0.248 . _ 0.523 0.363 0.040 0.693 07 0.168 72 1.204 100 1.218 101 2.178 00 2.087 85 4.684 02 4.700 02 - @.e20 8.700 -08 08 IS Trlal2 I P AA Trlal 1 19 Trlal 2 -1111 *BB NO ND NO NO 73 0.421 84 78 88 84 72. 73. 85 88 -100 00 03 - 9 0.388 78 0.884 88 1.116 111 1.041 07 2.141 107 4.801 82 4.394 an - - i0.001 100 10.402 105 96 10 PFOSA . .. -. . '! i '! I :. . BACK TO MAIN .. -. T:ilile 7: Suiitrilnry of Recoveries for Bl-FOS&-OII,Me-ITOSE-011,I'ITOS, I'OAA nr~tlYFOSA ill IIot Dog RoaQenl Blank B MalflX Blank A EAalflw Blenk B 0.5 ppb Spk A 0.6 ppb Spk 0 , 1.O ppb Spk C 2.0 ppb Spk E 6.0 ppb Spk Q 6.0 ppb Spk H 10.0 ppb Spk I 10.0 b S k J AVEnAOe - .. Et-FI Trlal 1 95 12 3 2w4 NO NO NO NO 0.488 0.385 1,004 1.027 1.000 1.080 4.950 6.340 - io.6oa 11.240 *89 I 07 73 Io0 103' 00 00 09 107 -108 112 rpo 10 --Mo-F Trial 1 ' -111 '9- 34L % NO m 0.307 NO 0.448 77 0.453 7 8 1.Do1 04 0.083 02 i.ie5 67 1.253 so 6.805 11; 1.380 11.462 Io.BBB 107 114 -108 90 21 Trlal 2 0.441 , 88 0.353 71 0.879 88 0.088. 07 1.820 , 08. 2.001 100 10.120 06 10 -,P. Trlnl 1 -111 'e Trlsl 2 11111/00 2 L L (R.c 0.178 P Trlsl 1 11/8/00 NO 0.070 0.173 0.857 87 0.640 84 1.041 92 1.137, 1or 1.087 03 2.008 00 6.248 103 6.1 17 10.080 - 0.812 100 -too 87 90 6 11.830 103 ' 106 0 6 AA Trlal2 flllllD0 10.428 97 B PI Trl.1 1 0.380 88 7 . Trlal2 I I I'npc 30 of 63 BACK TO MAIN ` h l h 8: Suttiitinryof Itccovcric for I'liOS, I'OAA :ti11 I'P0S.- in IliSll Raogonl Blank P P! Trhl 1 Trlrl2 112 1/00 -1li -0 2dlL if& 0.710 PI Trial 1 , 1A Trlal 2 * - 112 - 0 S!L ND ~ PI Trlal 1 llZl/OO sn]' -11; 2dlL E . ND Malrix Blank A Malilx Olank B 0.5 ppb. S p k A 0.5 ppb Spk B 1.0 ppb S p k C 1.0 ppb Spk D 2.0 ppb Spk E 2.0 P P S~pk F 5.0 ppb S p k G 5.0.ppb Spk,H 10.0 ppb Splc I .. AVERAGE: 115 26 Oi737 1.320 1.166 1.650 1.044 2.200 2.224 3.608 3.710 e.210 0.800 12.07~ 12.08: 61 120 102 00 113 123 00 315 -108 108 107 12 ND 4' ND NQ NQ - . NQ NQ 0.506 120 0.401 06 0.642 120 0.500 101 1.310 132. 1.003 100 1.300 140 1.116 112 2.884 143 2.141 107 .. 2.031 I.:& 1.892 9s 6.074 3.363 I 12.101 113 .^^ I40 121 11.140 4.942 9 0 5.701 114 11.43: 120 . I - 3 ., 108 ,130 117 6 11 9 , ' I -. .. .. P I 0 1 c 8 x F :? I :, . + CI 0 . 'E: c-i VI Y BACK TO MAIN I .. I BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 ....... .......... ... - . ._ - ._-.. _ _ - ---- .... ....... . -. .... .. Centre Protocol No. OOP-023-057 'FIGURES I ...... Ccnuc An;llytid L b O n t o r i ~ .In:. Study # 0 2 3 - W Centre Analytical Laboratories, Inc. .. ........... Centre Analytical Laboratories, Inc. .- -. ! I I. !. . I ._-__! .. . .Page33 of 69 . Page45 of 81 ....... Page 111 of 151 BACK TO MAIN .. .-... . ... .: .. . . . ........ .. .. ... . . ....... . . . . . . . . . . . . . . . .. Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 ................. .- . .. , -. -.. . . . . . . . . . ...._.. :. . ..... :: .'. . . _ :.. Centre Protocol No. oOP-023-057 ....;. ........ ....... - 1: ,. I, ..-. . .. 1 ___ _____ Centre Analytical Laboratories, Inc. . ! ! .! i page 34 Of 69 Page 46 of 81 Centre Analytical Laboratories, Inc.. __ . _. __-__ - -- - - - - . . ............ Page 112 of 151 .. ..... ... .-... BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-.057 ...... . ....... _. - ........... ..... Centre...Prot-o-c-o-l-N--o_. O_O_P_-023-057 I .. ! Centre Analytical Laboratories, Inc. - - - -- - -__ - - - ._ ._ Page 47 of 81 Page 113of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 .. . . - - . . . -. .- .. - . . .. .-. ---_ -. ..- - .. -- ----- F .. .. .. . . __ . -. - - ..-.-- .-. -'-le n tr . e Protocol No. OOP-023-057 --.__-------..- --__ .. I Figure 2: Representative Chromato,-; o f a Control Whole Apple sample (set ,111799Apple, Centre ID 983732 tipple Blank A) ! I cmrc h l y f i a l ' L b o n t o n ~I.nc. Study':: 02391 . I . Centre Analytical Laboratories, Inc. -. .... -- .- __.I ... ~ ' ' ... . . . .. _ . - - __- Centre Analytical Laboratories, Inc. Page 114 of 151 BACK TO MAIN . . . . . . . . . . . ......... ........... Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 .. ...... .. -- Centre Protocol No. COP-023-057 i Figure 2 (continued): Representative Chromatograms of a Control )no~le& l. ~ Sample (Set 111799 Apple, Centre ID 9 S X ? Apple Blank A) .. 4 : POXX 1983732Apple Blank A .. ._ .. .. I Centre Analytical Laboratories,Inc. __ -.- -.___ . .. - .......... Centre Analytical Laboratories, Inc. .. . . - .... ... ............... . . . . .- Page 49 of j.81 . Page 115 of 151 . . .. i . .. BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 -. .. ... -- Centre Protocol No. OOP-023-057 --. . .. ! - . ' l.bO ' 2.bo I 1 3.00 0 4.b0 ' 5.b 6.00 7 I . Centre Analytical Labol,+ories, Inc. Centre Analytical Laboratories, Inc. ,. ____......_ .... .- .. .. I ! ! ! i I -. Page 38 of 69 Page 50 of 8I Page 116 of 151 BACK TO MAIN __ ..................... Centre Study No.: 023-057 . Centre Protocol No: OOP-023-057 .. - - . . . . .- ..... .. .. .... ..... -. . . . . . -. -- . . . . . . ... . ----- .C. e. n. t.r.e.P.rotbcol NO. OOP-023-057 -2--- . . . I ! ! _. . Centre Analytical Liboratones, Inc. .- - - -- Centre Analytica1Laboratories,Inc. pig: 39 of 69 Page.51 of 81 Page 117 of 151 BACK TO MAIN ........... . . ...... Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 ................. . .-.-_.- . __-. . ...... .-... - ... Centre P r o t ~ o 1 . ~ . . ~ ~ ~ ~ 2 3 - 0 5 . 7 .. i LCIMSNS unit 6' MfiM of5Channels ES- 630 > 59 1.30e3 3: P?OS . bS08032 Green Eeans Blank A -2, 2137 .. i ! Centre Analytical Laboratories', IW.. -.- _ -.- -_.I _ __.._. . . ...... - ...... ~ __.._-.-. . .. ...... . Centre Analytical Laboratories, Inc. Page 40 of 69 Page 52 of 51 . . .. Page 118 of 151 BACK TO MAIN Centi-e Study No.: 023-057 Centre Protocol No: OOP-023-057 .. Centre Protocol No. 00P-023-057 . .- - . .. I .. I .?! l.bO " 2.b0 ' 3.b0 ' .. Area/ ------___- - . ...Centre Analytical Laboratories, Inc. . ...-. .. h$41 of 69 .I . . Page 53 of 81' Page 119 of 151 ............. . BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 , .. ............... . -.-. - --. - -. - - . ................... .. . . .. . . . .- ....-- _ _Centre Pro-.to..co--l-N-o-.-COP-023-057 1: E t - F O s i a 9908032 Spk B, Green Beans 500 ppt - . . . . I 1 . ............. Centre Analytical Laboratories; hc. _ _ .. ,. ,, . ._. Centre Analytical Laboratories,Inc. ..................... .. .. .. I Page 54 of 8 1 Page 120 of 151 ..... ............... BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 . ... . . _.. . ' Centre Analytical Laboratories,Inc.. I ,. . -..- -- ..-_. . ._.... -. -- .. . . . ... Centre Analytical Laboratories, Inc. ~ a g e 5 i O f ~. 1 - Page 121 of 151 ... - ......... i ' BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 -_ . . . . . -- .. .. ... - . --.-----_- .. . .........C..entre. P. r.ot.oc. o.l N..o..-...O. OP-623-057 ' ! - 2 : ne-?os-oE 912031 Egg BIankA . - . - 'i.2n2nCCS-1011 Srn (SG,2x2) - . 4 . 20.- 620 I .- 1.30e31 I. 3.09e4 Area ._ Centre Analytical Labora'tories. bc., .-. __ .-..___-. _ -. Centre Analytical Laboratories, Inc. __ - . . . . . . . .- . -. ......... ! Page 56 of 81; ' . . ..-__. Page 122 of 151 BACK TO MAIN ... .- I. Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 ........ ... . .. . . . . . . . . . Centre Protocol No. OOP-023-057 . -. ..i: . m990-1011 9912031 Lpg Bl& X 122?991011Srn (SG,2x2) . 5.87 1: . 29-Dec-1999 . .. of 5 Channels ES- . 4sa 78 .. .... .. C:me myrid~.,bo~toria~ncs~udya c r h o i ___ ----. ....... Centre Analytical Laboratories,Inc. ... ........... .---- .... ! ! ! . I i I --.I 45 of69 Page 57 of 81 . I .. - - .... Centre Analytical Laboratories, Inc. . ~ . ......._ ~ .. .. ...... Page 123 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023.057 I ._ .. r 1.f 1: 1" '.I .. . 7.18e31 Area I 6.58e4 - --- - , Centre Analytical Laboratories. Inc. '! I . ... . . . . -. .. ... .. . . .. .. I Centre Analytical Laboratories, Inc. . - . - -_-_. .......... Page 58 of 81 Page 124 of 151 ._..__ _.. .... .- - .- __ .. .. .. .. BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: 00P-023-057 .. . ----- . .... .. . -. .. . . . . . . ...'C.e. nt.re.P.rotocol No. COP-023-057 ~ I . .. .. "entre Analytical Labojtories, Inc. . . .. . . - . . I . ..._.. . - . .. ..-.. ..-...-. . Centre Analytical Laboratories, Inc. . - .-_.. page 47 of 69 Page 59 of 8I . Page 125 of 151 . . . .. . . . . . ...... .. .. BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: 00P-023-057 - -- Centre Protocol No. .@P:023r057--. l-. I 10 6 . E I: PCOS 310292 Chicken Ejlank B 11l,OSP612Sm (SG,2x2) ' 4.75 .- ~ 20-Nov-10(1903:41:4a . LCIMS/MS unit d . M%V af 5 Channeh ES- 49s > cc 420e3 I ---.. . . pentre Analytical Laboratories, Inc.' . . ..-.. - . . .- . . _..____.___C_....----. -. - .. Centre Analytical Laboratories, Inc. .--.- . - - I .--.-- - . _.... . ~ - -- - . P& 48 af 69 I Page 60 of 8 1 Page 126 of 151 . _ - . .. . . . - . - BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 - . . .. - . - - -..- .' .. Centre P r o t o c o f ~ . . ~ P - 0 2 ~ ~ 7 - - - - 1 . 1- 4.16e4 ATOa[ .. .. Centre Analytical Laboratories, In'c. -------_ .. _... . I - - _ _..... Page 49 of 69 Page61of81 . .. . . .. . Centre AnalyticalLaboratories,Inc. I--. - - __ __ Page 127 of 15 1 ... , . .. . , .. . BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: 00P-023-057 ! 1: Et-FOSz-oE . 9910292 Spk A, Chicken 500 ppt . . . .. - I .. Areal I -.._-_ _ _ _ _ _ _ . .. , GWCA n d y t i d hhntorio.Inc. Snrdy t 0 3 3 - ~ 1 . .. .. Centre Anhytical Laboratories. Inc. - .. . .- ~ . - .. -..--. Centre Analytical Laboratories, Inc. 50 of 69 Page 62 of 81 Page 128 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 . 559 1003 MRM of5Channels ES- 49a 78 9.Ei.4 __Centre Analytical.Labontorie;. Inc: -.---- . .- . . Centre Analytical Laboratories, Inc. ....- .. . Page63.of81 , -. . Page 129 of 151 .. I . I BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 . . Centre Protocol No.lX)P~023=05.7'-. 09-Nov-1s,o9 20:43:5$ MRM Of 5 Channels ES-1 u aI I 6.60 630 > 591 ---- Centre Analyticai Laoorat, L,!x. .. ....... ..... .. . . . - ._ Centre Analytical Laboratories, Inc. -- . ._ _ ~ __ . ........... ..... ... _I .... ! Page 130 of 151 ... .-......... BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 .. . .- . . . . . . . . . . . -- ........ . . . . . . . _.. . . . . . . .. . _. Centre prot&o&-IMEa't -057 1,-110999-2012Sm (SG,2x2) 1. . 4.07 9024 .. *- 413 > 369 3.99e4 . Area Centre myrid ~b~uoriInoc.,Study 03CO1 . ! j of 69 1: .I Centre Analytical Laboratories,Inc. ! Page 65 of 81 . - .. - -__... . . . . . -. .... ., ,- Centre Analytical Laboratories, Inc. Page 131 of 151 .. --- . ........................................ ........................................... . -. BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 I 1: Et-XJOS2-m ' . 572627SpikeA, Milk 500 ppt . ........ ~.-. ... CenUc hdyrid I;lbonmrics, he. SNdy n' 023-Wl Centre Analytical L.aboratories,Inc. ... . . . . --- . ..... Centre Analytical Laboratories, Inc. . ._ . .. I ! ! 1. I I 1 Page 132 of 151 BACK TO MAIN . .. . Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 -. . - - -- .- .--_ .. Centre Protoco1,No.-00P-023-057..... .. .. . . .. '! Centre Analytical Laboratories, Inc. . I Centre Analytical Laboratories, Inc. , ! Page 67 of 8 1 Page 133 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol NO: OOP-023-057 Centre A n d y t h Laboratories, h c . . i .. . . '! ---. . ...- ..__I--- I . Centre Analytical Laboratories, Inc. 1 . .~-.-____..__-.I._. I . . Page65of51 .. Page 134 of 151 .. .. 1 . ... BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: 00P-023-057 I E,We 12 (continued): `Representative bomato,$pms Ofa Control Brmd sample .(Set 111999Bread, Centre ID9905031Bread Blank A) . 111929-511 Sm (SG,2Y2) I17 "7. 4.57 . LC/MSIMS unit ' : I I ! .. . ccXrc .41.?Iytiul Lbamorir &I Study G3-001 ! . I I Centre Anatyucal Lauorarories, hc. --.. . . __. . . 1. - . -. . . . . . Centre Analytical Laboratories, Inc. . .............................. ....... - ........ ..... .Page 69 of 81 ........ Page 135 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 . . . . ., ........ .............. . . . . . . ....... ... .. ........... ................ .. . . . . . . . . . . . -.... .-. . . . Centre Protocol kOap--.O2~7-,,. .. I ,1 ' . .. . . . . , . . . . , _ .,_.._.., . . . , . . . . ( . . . . , . .. . ( . . . ) . ) . I . _ 1.00 200 3.00 4.00 5.00 1 I;.- 12202 . . I616>59 3.37.4 . Area, 4.84 19.81 I "1I 1 . 2.05e41 Area/ ! ... !' -.. . . . . . . . . --_. ....... F?&reAnalytical hboktories; Inc. , .... .. ....... Centre Analytical Laboratories, Inc. ....... .. ._ . . . . ... pa& sa of 69 Page 70 of 81: Page 136 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 .. .. .. .. . . . - ............ ... I . . .- .. --- ..... ._.__- ' Centre Protocol No. OOP-023-057. i I 1 '. .I . Centre m y l i c ~~ltmnioria~. n c . ~ d8 y023-001 . I .- page 59 of 69 Centre Analytical Laboratories, h C . ------- . .~.. . . . . .... - . Page 71 of 81 --- .--...... Centre Analytical Laboratories, Inc. .............................. . . .-...... ........... - . Page 137 of 151 ....... BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 i Centre Analytical Laboratories, Inc. . - ------- . . - ___ - - . - - -- Centre Analytical Laboratories, Inc. - - - - - _ -_ _ _ - - _-. - . ... ..___. .. . .. . Page 60of 69 I Page i ~of 3.1. -- Page 138 of 151 BACK TO MAIN ....... ,. . . . ......... . Centre Study No.: 023-057 Centre Protocol No: 00P-023-057 __ . . ... . . . . . . . . . . . -. . .-........-..... ...-. . . ............. .. .- . . -.. ... . . . . . . . . . . ... . ... . . .. .. - .: centre Protocd~a-QO.E42323-57 -1 .. Figure 14 (continued): Representative C ~ ~ ~ ~ ~ O oWfaI CLonStrol Hot Dog Sample (Set 110899Hot Dog, Centre ID 9903035Hot Dog Blank B) .. .- . Centre Analytical Laboratories. Inc. . .--__.I. ....... - - ............ . - - -_I_ _ _ - Centre Analytical Laboratories,Inc. __ ~ I ... I ........ Page61 of69 . Page 73'0f81 . .. ....... Page 139 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 .. i a : 110899-1015 ' T e - : Hot Dcg 500 ppt SpikS B .. _. .---------- C a m Mpd kbontoricr. I n r Study X 023-001 . Centre Analytical Laboratories; Inc. -.-- .- .-.... . . ., , . . .. -. _____ Centre Analytical Laboratories, Inc. ! I f. I b e 62 of 69 I 1 Page 140 of 151 BACK TO MAIN .- . .. Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 . - .. . .... ... . .. __---- ,. .... .... Centre Protocol No.-WP-O23--05Z__._ .. I Figure 15 (continued): Representative C h r o m a t o m from the Injection of Control Hot Dog Fortified at 500ppt each of Et-FOSE-OH, Me-FOSE-OH, PFOS, POA.4, and PFOSA (Set 110899Hot Dog, Centre ID9908035Spk B) -- -- Centre Analytical Laboratories, Inc. . .. . . . - - . .. . -. ..-. - .. -. .. .---- .. - - ___ Page _ .. 141 of . .. 151 .. ........... . . : . .... .. I BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 .... ...... Centre Protocot SO.-OQP-O~MW--- M W : 012400-2011 . 9 9 x 0 3 2 &tCLs6 BI& .. . . I 31 1 .. 5.31e31 . Area ! !. .---- ..__-I--_....._ Centre Analytical Laboratories, Inc. . .. . -. ! .I .. ,* .......... : Page 76 of 81 ..... -__..._. . . Centre Analytical Laboratories, Inc. .. - -. ........ - .... - ........... .- . .... . .... Page 142 of 151 . . . . . ....... BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: 00P-023-057 -. . . - .. . . . .. _ ._......... . ~ . . . . . . . ! I . N-: Ol2400-101( . 9912032 S;k A, mtfish 500 1: PFOS . 9912032 Spk A, Catfish 500 ppt . 012-1014 Sm (SG.2Q) 24-Jan-2000 20:;3:11 LC/MShlS unit 6 MRM of 5 Channels ES- .. ... Centre Analytical, Laboratoriej, Inc. -.-- ; Centre Analytical Laboratories, Inc. .- .- Page 77 of 81 .. . . Page 143 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 - . . . . - . . . .. ..-.. . .- . . . - ... .. - .- . - .. .- ... . . . ... . Centre Protocol N.Q?Oqp,O~3~057-.-~ .. I I. (. ~- - .. ! 2. __- I Centre Analytical Laboratories. Inc. . .. i . . ._- -. . . .. ....._._... .-. Centre Analytical Laboratories,Inc. --- - - __ - -. ._- . - .. 66 of 69 I Page 78 of 81 . .- Page 144 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 ... ......... ... .- .Eentre Protocei-~e,-001r823.057--. .. .. i .. .- _ I Figure 18 (continued): RepresentativeChromatograms of a Control Pork&rucle ' Sample (Set 122199 Pork,CentreID9911753 Pork Blank B) 4: PFOOSX -9911783Pork Blank B I ! ! ! .-- ~ (. Centre Analytical Laboratories, Inc. . I. . . . __....... .-..--- ..... - CentreAnalytical Laboratories,Inc. __-_. ... . . .. . ......, . ~ .... Page 79 of 81, . Page 145 of 151 .. .. BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 ,- . .. Centre Protocol N~.-WP-023,0.5J __ N-: 122139-2s . Tax:: 9911783 spk 9, Pork 530 P P C 1221SS215Sm ( S G , W ) .. . LC/MS/MSunit MRM of S Channels 3- 4528 630 > 5-0 4.17eS l 3: ??OS IS911783Spk B,Pork 500 PP* I 523 196iJ1 1.. 21-Dec-issg ;s:o?:34 : - __ - Centre Analytical Laboratories, Inc. _ . - .- ' P. a.ge 80 of 81 Page 146 of 151 BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 .. . ..... ~ . . ' . .' . ..... . Centre Protocol No. KIP-023-057 I 4 : 'OPOSX .. 9911783Spk E,Pork500 ppt ' 3.b0 - " 4.b0 I' - a 5.00- I- 6.b . 1S674 12A0reeila , Centre Analytical Laboratories. Inc. Centre Analytical Laboratories, Inc. Page 69 of 69 . Phge 81 of 81 Page 147 of 151 BACK TO MAIN Centre Study No.: 023-05L1.Centre Protocol No: 00P-023-057 CEntrE Analytical Laboratorks, Inc. 3048 Research Drive phone: (814) 231-8032 State College, PA 16801 Fax: (814) 231-1253 or . . (814) 231-15- 8_0- Page 1 of PROTOCOL A . ? V l E ~ M E ~ Amendment Number: 1 EffectiveDate: March 26,2001 Centre Study Number: 023-057 Centre Protocol Number: OOP-023-057 DESCRTPTION OF AMENDED SECTION 1. Page 5 of Protocol: Assips Susan Beach as the StudyDirector at 3M. 1. Change the study director to Karen Smith at Centre Anidyt.iCd Laboratories and make Susan Beach the Sponsor Representative at 3M. RATIONALE .. Reassignment of responsibi1ities for study at the request of Sponsor. IMPACT ON THE STUDY . There is no negative impact on the study. presentative Signature - Date Centre Analytical Laboratories, Inc. Page 148 of 151 ~ .. ,.. . _ .... . . ...... . ~~ ~~ BAC~ K TO MAIN .... . , - .. Centre Study No.: 023-057 a\ Centre Protocol No: OOP-023-057 C m t r r Analytical Laboratorks, Inc. 3048 Research Drive State College, PA I 6801 - Phone: (814) 231-8032 Fax: (814) 231-1253 or (814) 231-1580 Page 1 of 1 PROTOCOL AMENDMENT Amendment Number: 2 EffectiveDate: April 30.2001 Centre Study Number: 023-057 Centre Protocol Number: OOP-023-057 DESCRIPTION OF AMENDED SECTION 1. Page 7 of Protocol-Experimental Design: States that each set Of samples analyzed will contain at least one matrix coilti01and two fcrtificaticns. AMENDEQ! . 1. Add that each set of samples analyzed will also contain a reagent blank. 1. Added at request of the sponsor. RATIONALE W A C T ON THE STUDY 1. There is no negative impact on the study. I & V Sponsor Representative Signature b Date - Date CAL QAU Review .Centre Analytical Laboratories, Inc. Page 149of 151 BACK TO MAIN 3048 Research Drive State College, PA 16801 W&w.centrelab.com Phone: (814) 231-8032 (814) 231-1253 or (814) 231-1580 P Page 1 of 1 PROTOCOL A M E " E N T Amendment Number: 3 Effective Date: June 15,2001 Centre Study Number: 023-057 Centre Protocol Number: OOP-023-057 1. Study Title DESCRIPTION OF AMENDED SECTION AMENDEDB 1. Analysis of PFOS, FOSA and PFOA From Various Food Matrices Using HPLC Electrospray/Mass Spectrometry. RATIONALE 1. Changed for clarity at the request of the sponsor. IMPACT ON THE STUDY 1. There is no negative impact on the study. Centre Analytical Laboratories, Inc. Page 150 of 151 .__ BACK TO MAIN Centre Study No.: 023-057 Centre Protocol No: OOP-023-057 CEntre Analytical Laboratories, Inc. 3048 Research Drive State College, PA I 6801 phone: (814) 231-8032 Fax: (814) 231-1253 or (814) 231-1580 PROTOCOL DEVIATION Page'l of Deviation Number: 1 Date of Occurrence: 1) January 19,2001; 2) February 15,2001 Centre Study Number: 023-057 Sponsor Study Number: NA DESCRIPTION OF DEVIATION 1) A recovery of 56% for FOSA is being accepted for the 10.0n d g Spk B in data set 011901A Fish. 2) A CCV recovery oi 143% is being accepted for the 2.5 nd/mL standard for FOSA in data set 021401A Beef. ACTIONS TAKEN i.e.. amendment issued, SOP revision. etc... 1) Deviation issued. 2) Deviation issued. *lizh - tecorded ByiDate: U IMPACT ON THE STUDY 30 1) There was no negative impact on the study since the 2.5 n g h d Spk A for FOSA in lata set 011901A Fish was within the 60-130%acceptable range and the levels of residue found in the fish samples were: all below 2.5 ng/pd?@ @@ky 2) There was no negative impact on the study since the CCV'S before and the C C v run after the standard in question were acceptable. / gncipal u t i g a t o r Signature Date FJk c kudy Director Signature 3-?-/, s- ponsor Manazement Signature Date WA,A Date Centre QAU Review 6 hh, Centre Analytical Laboratories, Inc. Page 151of 151