Document 9JXj4ZONJ62zV411R9GGEq70R

Project / Study Archive Request 1 ReauestorName Kent Lindstrom Report I SMCA areas for Archivist use onlv 1 I I Phone 651 778-5352 I I Date 09122106 I n 1 Reauestfor: Archival I Retrieval FC Title/Summary: Interim Report #24 -Analysis of Water, Sludge, and Sediment Samples Study Title: Analysis of Perfluorooctanoic Acid (PFOA) in Water, Soil, Sediment, Fish, Clams, Vegetation, Small Mammal Liver and Small Mammal Serum Using LC/MS/MS for the 3M Decatur Monitoring Program Executive Summary GrouD: 3M Decatur Group Project Number: Exygen Protocol Number PO000760 Project Lead/ Study Director: Gary Hohenstein (data management), Mike Santoro (Sponsor), John Flaherty (Principal Investigator), Jaisimha Kesari P.E., DEE (Study Director, Weston Solutions, Inc) Contract: Exygen Research Contract Proiect #: PO000760 Analyte(s): PFOA Test System (Sample Matrix): Water, Sludge, Sediment Project Type I Analysis 0Degradation 0Duct Residue Analysis II Ecotox u Regulatory Status: Non-GLP GLP I [7Method Validation I 0Phys/Chem Properties 0ResearchMethod Development I Other: Initiation Date: 11/05/04 (Protocol Signed by Study Director), Analytical Start Date for Interim Report: 05/09/06 Completion Date: Analytical Termination Date for Interim Report #24: 07/07/06 Signed by Study Director: See Report, Signed by Sponsor: See Report 0 m m Electronic Media: Data CD: Exygen) FTIR Data Page 1 of2 Project / Study Archive Request Archive Date Scan Date W-Site Storage Date Key Words: Decatur Memorandum of Understanding, MOU, GLP, Protocol P0000760, PFOA, E05-0209, E05-0210, Interim Report #24 Comments: Note to Archivist. This is Interim Report #24 for this LlMS project (GLP). There will be multiple Interim reports. Please include the Interim Report number in the title of the PDF. Please send this entire report including the raw data via web to Jai Kesari. Please send a copy of the report without raw data to Michael Santoro and to Gary Hohenstein. Distribution Point Gary Hohenstein (report without raw data) Jai Kesari (report with raw data) Michael Santoro (report without raw data) Date - 09/22/06 - 09/22/06 - 09/22/06 Page 2 of 2 INTERIM REPORT #24 - Analysis of Water, Sludge, and Sediment Samples STUDY TITLE Analysis of PerfluorooctanoicAcid (PFOA) in Water, Soil, Sediment, Fish, Clams, Vegetation, Small Mammal Liver and Small Mammal Serum Using LC/MS/MS for the 3M Decatur Monitoring Program DATA REQUIREMENTS EPA TSCA Good Laboratory Practice Standards 40 CFR 792 STUDY DIRECTOR Jaisimha Kesari P.E., DEE Weston Solutions, Inc. 1400 Weston Way West Chester, PA 19380 Phone: 610-701-3761 INTERIM REPORT COMPLETION DATE September 13,2006 PERFORMING LABORATORY Exygen Research 3058 Research Drive State College, PA 16801 Phone: 814-272-1039 STUDY SPONSOR 3M Company 3M Building 0236-01-B-10 St. Paul, MN 55144 Phone: 651-733-6374 PROJECT Protocol Number: PO000760 Exygen Study Number: PO000760 Total Pages: 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT Exygen Study Number P0000760, entitled "Analysis of Perfluorooctanoic Acid (PFOA) in Water, Soil, Sediment, Fish, Clams, Vegetation, Small Mammal Liver and Small Mammal Serum Using LC/MS/MS for the 3M Decatur Monitoring Program," conducted for 3M Company, is being performed in compliance with EPA TSCA Good Laboratory Practice Standards 40 CFR 792 by Exygen Research. John Flahdy Principal Investigator Exygen Research Study Director Weston Solutions, Inc. + Michael A. S Sponsor Representative 3M Company Exygen Research Page 2 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 QUALITY ASSURANCE STATEMENT Exygen Research's Quality Assurance Unit reviewed Exygen Study Number P0000760, entitled, "Analysis of Perfluorooctanoic Acid (PFOA) in Water, Soil, Sediment, Fish, Clams, Vegetation, Small Mammal Liver and Small Mammal Serum Using LC/MS/MS for the 3M Decatur Monitoring Program". All reviewed phases' were inspected for conduct according to Exygen Research's Standard Operating Procedures, the Study Protocol, and all applicable Good Laboratory Practice Standards. All findings were reported to the Exygen Principal Investigator and Management and to the Study Director. Phase 34) Raw Data and Interim Report Review 36) Raw Data and Interim Report Review Date Inspected 6126-27106 Date Reported to Date Reported to Principal Exygen Date Reported to Investigator Management Study Director 07/06/06 07/07/06 07107/06 09/12/06 0911 3/06 09113/06 09113/06 Date `Note: All in-lab inspections will be documented in the QA statement for the final analytical report at the conclusion of the study. This QA statement involves only the review of the interim report and associated raw data. Exygen Research Page 3 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 CERTIFICATION OF AUTHENTICITY This interim report, for Exygen Study Number P0000760, is a true and complete representation of the raw data. Submitted by: Exygen Research 3058 Research Drive State College, PA 16801 (814) 272-1039 Principal Investigator, Exygen: John Flahedfy Vice President Exygen Research Exygen Research Facility Management: President Study DireHor, Weston Solutions, Inc. Jaisimha"Kesari P.E., DEE Weston Solutions, Inc. Sponsor Representative, 3M Company: Michael A. Sftom Director of egulatoryAffairs Exygen Research Page 4 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 STUDY IDENTIFICATION Analysis of PerfluorooctanoicAcid (PFOA) in Water, Soil, Sediment, Fish, Clams, Vegetation, Small Mammal Liver and Small Mammal Serum Using LC/MS/MS for the 3M Decatur Monitoring Program PROTOCOL NUMBER: PO000760 EXYGEN STUDY NUMBER: PO000760 TYPE OF STUDY: Residue SAMPLE MATRIX: Water, Sludge, and Sediment TEST SUBSTANCE: Perfluorooctanoic acid (PFOA) SPONSOR: 3M Company 3M Building 0236-01-B-10 St. Paul, MN 55144 STUDY DIRECTOR: Jaisimha Kesari P.E., DEE Weston Solutions, Inc. 1400 Weston Way West Chester, PA 19380 STUDY MONITOR: Michael A. Santoro 3M Company 3M Building 0236-01-B-10 St. Paul, MN 55144 PERFORMING LABORATORY: Exygen Research 3058 Research Drive State College, PA 16801 ANALYTICAL PHASE TIMETABLE: Study Initiation Date: 11/05/04 Interim Analytical Start Date: 05/09/06 Interim Analytical Termination Date: 07/06/06 Interim Report Completion Date: 09/13/06 Exygen Research Page 5 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 PROJECT PERSONNEL The Study Director for this project is Jaisimha Kesari at Weston Solutions, Inc. The following personnel from Exygen Research were associated with various phases of this interim portion of the study: Name John Flaherty Karen Risha Chrissy Edwards Mark Ammerman Amy Sheehan Eric Edwards Mindy Cressley Brian McAllister Frances Crespi Sharareh Zolghadr Scott Crain Krista Gallant Brittany Kravets Kim Hall Vice President Laboratory Supervisor Technician Sample Custodian Associate Scientist Sample Custodian Technician Sample Custodian Technician Technician Technician Technician Technician Technician Exygen Research Page 6 of 140 Interim Report #24 . Analysis of Water. Sludge. and Sediment Exygen Study No .. PO000760 TABLE OF CONTENTS PaRe TITLE PAGE ....................................................................................................................... 1 GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT.............................. 2 QUALITY ASSURANCE STATEMENT .......................................................................... 3 CERTIFICATION OF AUTHENTICITY ........................................................................... 4 STUDY IDENTIFICATION................................................................................................ 5 PROJECT PERSONNEL..................................................................................................... 6 TABLE OF CONTENTS..................................................................................................... 7 LIST OF TABLES ............................................................................................................... 8 LIST OF FIGURES.............................................................................................................. 9 LIST OF APPENDICES .................................................................................................... 10 1.0 SUMMARY................................................................................................................ 11 2.0 OBJECTIVE ............................................................................................................... 12 3.0 INTRODUCTION....................................................................................................... 12 4.0 ANALYTICAL TEST SAMPLES.............................................................................. 12 5.0 REFERENCEMATERIAL........................................................................................ 12 6.0 DESCRIPTION OF ANALYTICAL METHOD ........................................................ 13 6.1 Extraction Procedure for Water ................................................................................ 14 6.2 Extraction Procedure for Sludge............................................................................... 14 6.3 Extraction Procedure for Sediment........................................................................... 14 6.4 Percent Solids DeterminationFor Sludge and Sediment.......................................... 15 6.5 Preparation of Standards and Fortification Solutions............................................... 15 6.6 Chromatography....................................................................................................... 16 6.7 Instrument Sensitivity............................................................................................... 17 6.8 Description of LC/MS/MS Instrument and Operating Conditions........................... 17 6.9 Quantitation and Example Calculation..................................................................... 18 7.0 EXPERIMENTAL DESIGN ...................................................................................... 21 8.0 RESULTS ................................................................................................................... 22 9.0 CONCLUSIONS......................................................................................................... 23 10.0 RETENTION OF DATA AND SAMPLES ............................................................. 24 Exygen Research Page 7 of 140 Interim Report #24 . Analysis of Water. Sludge. and Sediment Exygen Study .No. PO000760 Table I. LIST OF TABLES PaJg Summaryof PFOA in Water Samples............................................................ 26 Table 11. Summary of PFOA in Re-extracted Water Samples....................................... 29 Table III. Summaryof PFOA in Sludge Samples........................................................... 30 Table IV. Summary of PFOA in Sediment Samples....................................................... 31 Table V. Matrix Spike Recovery of PFOA in Water Samples...................................... 32 Table VI. Matrix Spike Recovery PFOA in Re-extracted Water Samples..................... 39 Table VII. Matrix Spike Recovery of PFOA in Sludge Samples..................................... 40 Table VIII. Matrix Spike Recovery PFOA in Sediment Samples..................................... 41 Table IX. Surrogate Spike Recovery of I3CPFOA in Water Samples ........................... 42 Table X. Surrogate Spike Recovery of I3CPFOA in Re-extracted Water Samples......49 Table XI. Surrogate Spike Recovery of I3CPFOA in Sludge Samples.......................... 50 Table XII. Surrogate Spike Recovery of 13CPFOA in Sediment Samples...................... 51 Table XIII. Total Percent Solids for Sludge and Sediment Samples................................. 52 Exygen Research Page 8 of 140 ~~ ~~ ~ Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Figure 1. LIST OF FIGURES Paae Typical Calibration Curve for PFOA in Reagent Water.................................54 Figure 2. Extracted Standards of PFOA in Reagent Water, 25 ng/L and 50 ng/L, Respectively....................................................................................................55 Figure 3. PFOA in Reagent Control, 50 ng/L Fortified Reagent Spike A, and 500 ng/L Fortified Reagent Spike B, Respectively.......,................................................56 Figure 4. Chromatogram Representing a Water Sample Analyzed for PFOA (Exygen ID: C0169336, Data Set: 042806B) ................................................. 57 Figure 5. ChromatogramRepresenting a Sludge Sample Analyzed for PFOA (Exygen ID: C0171106,Data Set: 050906BR)............................................... 58 Figure 6. Chromatogram Representing a Sediment Sample Analyzed for PFOA (Exygen ID: C0171108, Data Set: 051106A)................................................. 59 Figure 7. Typical Calibration Curve for I3CPFOA in Reagent Water .......................... 60 Figure 8. Figure 9. Extracted Standards of 13CPFOA in Reagent Water, 25 ng/L and 50 ng/L, Respectively.............,...,...................................................................61 13C PFOA in Reagent Control, 50 ng/L Fortified Reagent Spike A, and 500 ng/L Fortified Reagent Spike ByRespectively.................................. 62 Figure 10. Chromatogram Representing a Water Sample Analyzed for I3CPFOA (Exygen ID: C0169336, Data Set: 042806B) ................................................. 63 Figure 11. ChromatogramRepresenting a Sludge Sample Analyzed for I3CPFOA (Exygen ID: C0171106, Data Set: 050906B) ................................................. 64 Figure 12. ChromatogramRepresenting a Sediment Sample Analyzed for I3CPFOA (Exygen ID: C0171108,Data Set: 051106A)................................................. 65 Exygen Research Page 9 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 LIST OF APPENDICES Paae Appendix A Study Protocol PO000760 (Exygen Study No. P0000760) with Analytical Methods and Protocol Amendments ........................................66 Exygen Research Page 10 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 1.0 SUMMARY Exygen Research extracted and analyzed water, sludge, and sediment samples for the determination of perfluorooctanoic acid (PFOA) according to Exygen Methods VOOOl780, VOOOl781, and V0001782, respectively (Appendix A). The limit of quantitation for PFOA in water was 25 ng/L. The limit of quantitation for PFOA in sludge and sediment was 0.2 ng/g (wet weight). Analytical results for the analysis of PFOA in water samples are summarized in Table I. Analytical results for the analysis of PFOA in re-extracted water samples are summarized in Table 11. Analytical results for the analysis of PFOA in sludge samples are summarized in Table 111. Analytical results for the analysis of PFOA in sediment samples are summarized in Table IV. Quantitative results were obtained for all samples and analytes except for samples from sites DAL-GW-138R-0-060412, DAL-GW-138S-O060412, DAL-SD-OSPO1-0-0000, and DAL-SD-OSP02-0-0000 that are not reported (NR) due to quality control failures. Fortification recoveries for PFOA in the water samples are detailed in Table V. The average percent recovery k standard deviation for PFOA in the water samples was 75 f 22%. Fortification recoveries for PFOA in the re-extracted water samples are detailed in Table VI. The average percent recovery f standard deviation for PFOA in the reextracted water samples was 99 f 16%. Fortification recoveries for PFOA in the sludge samples are detailed in Table VII. The average percent recovery for PFOA in the sludge sample was 69%. A standard deviation was not calculated for PFOA in sludge because only one sample was analyzed. Fortification recoveries for PFOA in the sediment samples are listed as NR in Table VIII. The average percent recovery k standard deviation for PFOA in the sediment samples was not calculated due to quality control failures. Fortification recoveries for I3CPFOA in the water samples are detailed in Table IX. The average percent recovery & standard deviation for 13CPFOA in the water samples was 75 f 26%. Fortification recoveries for 13C PFOA in the re-extracted water samples are detailed in Table X. The average percent recovery f standard deviation for 13CPFOA in the re-extracted water samples was 97 k 34%. Fortification recoveries for 13CPFOA in the sludge samples are detailed in Table XI. The average percent recovery k standard deviation for 13CPFOA in the sludge samples was 49 f 16%. Fortification recoveries for 13CPFOA in the sediment samples are listed as NR in Table X I . The average percent recovery f standard deviation for I3C PFOA in the sediment samples was not calculated due to quality control failures. Percent recoveries have not been reported for samples that contained analyte levels three times or greater than the fortification level. Percent solids for sludge and sediment samples are detailed in Table XIII. Exygen Research Page 11 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.:PO000760 2.0 OBJECTIVE The objective of the analytical part of this study was to determine level of perfluorooctanoic acid (PFOA) in water, sludge, and sediment samples according to Protocol PO000760 (Appendix A). 3.0 INTRODUCTION This report details the results of the analysis for the determination of PFOA, in water samples using the analytical method entitled, "V0001780: Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS," in sludge using the analytical method entitled, "VOOO1781: Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS," and in sediment using the analytical method entitled, "V0001782: Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Sediment by LC/MS/MS." The study was initiated on November 05, 2004, when the study director signed protocol number P0000760. The analytical start date for this interim report was May 9, 2006 and the analytical termination date for this interim report was July 07,2006. 4.0 ANALYTICAL TEST SAMPLES In total, one hundred and sixty water samples, one sludge sample, and two sediment samples were received from the client. Thirty-eight water samples (Exygen ID: C0169333 - C0169370) were received on wet ice on March 13,2006 from Tim Frinak at Weston Solutions, Inc. One hundred and twenty-two water samples, one sludge sample, and two sediment samples (Exygen ID: C0170984 - C0171108) were received on wet ice on March 18, 2006 from Tim Frinak at Weston Solutions, Inc. All samples were logged in by Exygen personnel and placed in refrigerated storage. Sample log-in and chain of custody information is located in the raw data package associated with this interim report. Storage records will be kept at Exygen Research. 5.0 REFERENCE MATERIAL The analytical standard, PFOA, was purchased from Oakwood Products, Inc. and was received at Exygen on March 17, 2005. The surrogate spiking standard, 13C labeled perfluorooctanoic acid (I3C PFOA SP0004184), was received at Exygen on April 15, 2004 from the 3M Company. Exygen Research Page 12 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 The available information for the reference materials is listed below. PFOA was stored ambient and 13CPFOA was stored frozen. Compound PFOA 13CPFOA Exygen Inventory No. SP0005444 SP0004184 Lot # 203 3507-195 Purity (%) Expiration Date 99 0313 1/07 97 03/29/09 The molecular structures of PFOA and 13CPFOA are given on the following pages: PFOA Chemical Name: Perfluorooctanoic acid Molecular Weight: 414 Transitions Monitored: 413 + 369 (for quantification) and 413 -+219 (for confirmation) Structure: 13C PFOA Chemical Name: 1,2-13C perfluorooctanoic acid Molecular Weight: 416 Transition Monitored: 415 + 370 Structure: F F F I F 6.0 DESCRIPTION OF ANALYTICAL METHOD The analytical methods "V0001780: Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Water by LCMSMS," "V0001781: Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Soil by LCMSMS," and "V0001782: Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Sediment by L C M S M S " were used for this study. Exygen Research Page 13 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 6.1 Extraction Procedure for Water A 40 mL aliquot of the water sample was used for the extraction procedure. After fortification of appropriate samples, the samples were loaded onto a CISSPE cartridge conditioned with 10 mL of methanol and 5 mL of water. The eluate was discarded. Approximately five milliliters of methanol was added to the cartridge. Five milliliters of eluate was collected into a graduated 15 mL polypropylene centrifuge tube. Each sample was analyzed by LC/MS/MS electrospray. 6.2 Extraction Procedure for Sludge Before the samples were weighed for the extraction, they were mixed thoroughly by vigorous shaking the sample container. A 5-gram portion of sludge was weighed into a fifty-milliliter centrifuge tube for the extraction. After fortification of appropriate samples, 5 mL of methanol was added to the samples. The samples were allowed to shake on a wrist action shaker for -15 minutes and were then sonicated in an ultrasonic bath for -15 minutes. The volume was taken to 40 mL with water and the samples were then centrifuged for -10 minutes at -3000 rpm. The supernatant was then loaded onto a CIS SPE cartridge conditioned with 10 mL of methanol and 5 mL of water. The eluate was discarded. Approximately five milliliters of methanol was added to the cartridge. Five milliliters of eluate was collected into a graduated 15 mL polypropylene centrifuge tube. Each sample was analyzed by LC/MS/MS electrospray. 6.3 Extraction Procedure for Sediment Before the samples were weighed for the extraction, they were mixed thoroughly by vigorously shaking the container. A 5-gram portion of sediment was weighed into a fiftymilliliter centrifuge tube for the extraction. After fortification of appropriate samples, 35 mL of 1% acetic acid in water was added to the samples. The samples were vortexed and allowed to shake on a wrist action shaker for -60 minutes. The samples were centrifuged for -20 minutes at -3000 rpm. The supernatant was then loaded onto a CISSPE cartridge conditioned with 10 mL of methanol and 20 mL of water. The eluate was discarded. Twenty milliliters of methanol was added to the sediment samples left in the centrifuge tube. The samples were vortexed and allowed to shake on a wrist action shaker for another 30 minutes. The samples were centrifuged again for -20 minutes at -3000 rpm. The supernatant was then loaded onto the same CIS SPE cartridge. The eluate was collected into a 500 mL Nalgene Bottle. The column was washed with 4 mL of methanol. The wash was collected in the same bottle as the eluate. Approximately two hundred milliliters of water was added to the bottles. The samples were mixed by shaking and loaded onto another c18 SPE cartridge conditioned with 10 mL of methanol and 20 mL of water. The eluate was discarded. Approximately five milliliters of methanol was added to the cartridge. Five milliliters of eluate was collected into a Exygen Research Page 14 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 graduated 15 mL polypropylene centrifuge tube. Each sample was analyzed by LC/MS/MS electrospray. 6.4 Percent Solids Determination For Sludge and Sediment Percent solids were determined using the procedure indicated in Exygen method V0000427. Approximately 20 grams of sample was weighed into a pan. The weight of the sample plus the pan was recorded. The sample was then dried in an oven overnight at 104 f2 "C.Then the sample was transferred to a dessicator and allowed to cool for -15 minutes. Each sample was then weighed again, including the weight of the pan. The percent solid for each sample was then calculated. 6.5 Preparation of Standards and Fortification Solutions A mixed stock standard solution of PFOA and I3C PFOA was prepared at a concentration of 1000 pg/mL by dissolving 100 mg of each of the standards (corrected for purity and salt content) in 100 mL of methanol. From this solution, the following fortification standards were prepared: Conc. of Stock or Fort. Fort Volume Solution (pg/mL)' (mL) 1000 10 100 10 10 10 1.o 10 0.1 10 of PFOA and 13CPFOA Final Volume (mL) 100 100 100 100 100 Final Conc. of Fortification Std. (PdrnL) 100 10 1.o 0.1 0.01 A stock solution of only 13CPFOA was also prepared at a concentration of 100 pg/mL by dissolving 10 mg the standard (corrected for purity and salt content) in 100 mL of methanol. From this solution, the following fortification standards were prepared: Conc. of Stock or Fort. Solution (pg/mL)' 100 10 1.o o P 3 cPFOA Fort Volume (mu 10 10 10 Final Volume (mu 100 100 100 Final Conc. of Fortification Std. (Pg/mL) 10 1.o 0.1 Exygen Research Page 15 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 A set of non-extracted calibration standards containing PFOA and I3C PFOA (for the sediment samples) was prepared in methanol, as specified in Exygen method V0001782. The following concentrations were prepared: Conc. of Fort Fort Solution Volume (ng/mL)' (mL) 100 1 100 0.5 100 0.2 10 1 5 1 2 1 of PFOA and I3CPFOA Volume of Fortified Sample (mL) 10 10 10 10 10 10 Final Conc. of Calibration Std. (ng/mL) 10.0 5.0 2.0 1.o 0.5 0.2 A set of extracted calibration standards containing PFOA and I3C PFOA (for the sludge and water samples) was prepared in water before extraction, as specified in Exygen methods VOOOl780 and VOOOl781. Conc. of Fort Solution Fort Volume (ng/mL)' (mL) - - 10 0.1 10 0.2 10 0.4 100 0.1 100 0.2 100 0.4 ofPFOA and I3CPFOA Volume of Fortified Sample (mL) 40 40 40 40 40 40 40 Final Conc. of Calibration Std. (ng/L) 0 25 50 100 250 500 1000 The stock standard solution and all fortification and calibration standard solutions were stored in a refrigerator (4" k 2C) when not in use. Documentation of standard preparation is located in the raw data package associated with this interim report. 6.6 Chromatography Quantification of PFOA and I3C PFOA was accomplished by LC/MS/MS electrospray. The retention time of PFOA and I3C PFOA was -1 1.5 min. Peaks above the LOQ were not detected in any of the reagent blank samples corresponding to the analyte retention time. Exygen Research Page 16 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 6.7 Instrument Sensitivity The smallest standard amount injected during the chromatographic run had a concentration of 25 ng/L of PFOA and I3C PFOA for water samples and a concentration of 0.2 ng/mL of PFOA and I3CPFOA for sludge and sediment samples. 6.8 Description of LC/MS/MS Instrument and Operating Conditions Instrument: Interface: Computer: Software: HPLC: API 4000 Biomolecular Mass Analyzer Turbo Ion Spray Liquid Introduction Interface DELL OptiPlex GX400 Windows NT, Analyst 1.4.1 Hewlett Packard (HP) Series 1100 HP Quat Pump HP Vacuum Degasser HP Autosampler HP Column Oven HPLC Column: ThennoFluophase Rp,50 mm x 2.1 mm Column Temp.: 30" C Injection Vol.: 15 pL Mobile Phase (A): 2 mM Ammonium Acetate in water Mobile Phase (B): Methanol Time (min) %A %B 0.0 65 35 1.o 65 35 8.0 25 75 10.0 25 75 11.0 65 35 18.0 65 35 Total run time: -18 min Flow Rate: 0.3 mL/min Ions monitored: Analyte PFOA PFOA Confirm Ion I3C PFOA Mode negative negative negative Transition Monitored 413 -+ 369 413 + 219 415 + 370 Approximate Retention Time (min) -1 1.5 min. -1 1.5 min. -1 1.5 min. Exygen Research Page 17 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 6.9 Quantitation and Example Calculation Fifteen microliters of sample or calibration standard were injected into the LCRLISMS. The peak area was measured and the standard curve was generated (using l/x fit weighted linear regression) by Analyst software using six concentrations of standards. The concentration was determined from the equations below. For water and sludge samples: Equation 1 calculated the amount of analyte found in sludge and water (in ng/L, based on peak area) using the standard curve (linear regression parameters) generated by the Analyst software program. Equation 1: Analyte found (ng/L) = [Peak area - intercept) x DF slope Where: DF = Dilution Factor, factor by which the final volume was diluted, if necessary. For samples fortified with known amounts of PFOA and I3C PFOA prior to extraction, Equation 2 was used to calculate the percent recovery. Equation 2: Recovery (%) = [analyte found ( n a ) - analyte in control ( n a ) ) x100% amount added (ng/L) Note: For the PFOA recovery calculation, the "control" is the unspiked aliquot of the primary field sample. Equation 3 was used to convert the amount of sludge PFOA found in ng/L to ng/g (ppb). Equation 3: PFOA found (ppb) = [PFOA found (ndL) x volume extracted (0.04L)I sample weight (5 g) Equation 4 was then used to calculate the amount of PFOA found in ppb based on dry weight. Equation 4: PFOA found (ppb) dry weight = PFOA found (ppb) x [loo% / total solids(%)] NOTE: Total solids (%) = [wet weight (8) / dry weight (g)] x 100% Exygen Research Page 18 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 An example of a calculation using an actual sample follows: Sludge sample Exygen ID C0171106 Spk D (Set: 050906B), fortified at 50,000 ng/L with PFOA where: peak area intercept slope dilution factor ng/L PFOA added (fort level) - 406453 - 6660 - 456 - - 100 -- 50,000 ng/L in corresponding sample (ng/L) = 53000 volume extracted (L) sample weight (g) total solids (%) From equation 1 : Analyte found (ng/L) = J406453- 66601 x 100 456 = 87700ndL From equation 2: % Recovery = (87700 ng/L - 53000 ng/L) x 100% 50000 ng/L = 69% From equation 3: PFOA found (ppb) = (87700 n d L x 0.04L) 5g = 702ppb From equation 4: PFOA found (ppb) dry weight = 702 ppb x (100% / 22.83%) = 3070ppb Note: Numbers may vary slightly due to rounding. For sediment samples: Equation 5 calculated the amount of analyte found in sediment (in ng/mL, based on peak area) using the standard curve (linear regression parameters) generated by the Analyst software program. Exygen Research Page 19 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Equation 5: Analyte found (ng/mL) = (Peak area - intercept) x DF slope Where: DF = Dilution Factor, factor by which the final volume was diluted, if necessary. For samples fortified with known amounts of PFOA and 13CPFOA prior to extraction, Equation 6 was used to calculate the percent recovery. Equation 6: Recovery (%) = Janalyte found (ndmL) - analyte in control (ndmL)) x 100% amount added (ng/mL) Note: For the PFOA recovery calculation, the "control" is the unspiked aliquot of the primary field sample. Equation 7 was used to convert the amount of sediment PFOA found in ng/mL to ng/g (PPb). Equation 7: PFOA found (ppb) = IPFOA found (ng/mL) x volume extracted ( 5 mL)] sample weight ( 5 g) Equation 8 was then used to calculate the amount of PFOA found in ppb based on dry weight. Equation 8: PFOA found (ppb) dry weight = PFOA found (ppb) x [loo% / total solids(%)] NOTE: Total solids (%) = [wet weight (g) / dry weight (g)] x 100% An example of a calculation using an actual sample follows: Sediment sample Exygen ID C0171107 Spk C (Set: 051106A), fortified at 0.5 ng/mL with PFOA where: peak area intercept slope dilution factor - 327495 - 1130 - 190000 - - 1 ng/mL PFOA added (fort level) - 4.0 ng/mL in corresponding sample (ng/mL) = volume extracted (L) - - sample weight (g) - - total solids (%) - - 0.440 5 5 58.39 Exygen Research Page 20 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 From equation 5: Analyte found (ng/mL) = 1327495 - 11301 190000 = 1.72ngh-d From equation 6: % Recovery = (1.72 ndmL - 0.440 ndmL) x 100% 4 ng/mL = 32% From equation 7: PFOA found (ppb) = (1.72 ndmL x 5 mL) 5g = 1.72 ppb From equation 8: PFOA found (ppb) dry weight = 1.72 ppb x (100% / 58.39%) = 2.95 ppb Note: Numbers may vary slightly due to rounding. 7.0 EXPERIMENTAL DESIGN I3C PFOA was used as a surrogate for all the samples except the rinse blanks. I3C PFOA was added to the sludge and sediment samples and sample replicates in the laboratory after collection. I3C PFOA was added to the water sample collection bottles in the laboratory before being shipped to the field for sampling. For water samples designated as field matrix spikes, PFOA was also added at a known concentration to the bottles in the laboratory before being shipped to the field. The water sample bottles were filled to a 200 mL volumetric fill line in the field except for C0169347 and C0169354, which were filled to 250 mL, and C0171088, which was filled to 230 mL. The water samples and trip blank samples were analyzed in thirteen sets. Each set included one reagent blank and two reagent blanks fortified at known concentrations. The first water set contained two sample sites and one trip blank and its associated spikes. Water sets two through twelve contained three sample sites each. The thirteenth water set contained one sample site and two trip blanks and their associated spikes. For each site, a sample, a field duplicate and two or three matrix field spikes were collected. For each site, a laboratory duplicate of the primary sample was extracted and two laboratory matrix Exygen Research Page 21 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 spikes were also extracted. For the two laboratory matrix spikes, two 40 mL portions of the primary sample collected for the site was poured from the bottle and fortified. Not only was PFOA added in the laboratory prior to extraction, but I3CPFOA was also added. The additional I3C PFOA was added because the levels of PFOA spiked into the samples were known to exceed the calibration ranges and were not analyzed without dilution; therefore, I3C PFOA levels were adjusted to require the same dilution as the other analytes. The re-extracted water samples were analyzed in three sets. Each set included one reagent blank and two reagent blanks fortified at known concentrations. The first set contained eighteen samples with appropriate spiking levels. The second set contained fourteen samples with appropriate spiking levels. The last set contained six samples and the appropriate spiking levels. For the two laboratory matrix spikes, two 40 mL portions of the primary sample collected for the site was poured from the bottle and fortified. Not only was PFOA added in the laboratory prior to extraction, but I3CPFOA was also added. The additional I3C PFOA was added because the levels of PFOA spiked into the samples were known to exceed the calibration ranges and were not analyzed without dilution; therefore, I3C PFOA levels were adjusted to require the same dilution as the other analytes. The single sludge sample was extracted in one set. The set included one reagent blank and two reagent blanks fortified at known concentrations. For each sample, a laboratory duplicate of the sample and two laboratory matrix spikes were also extracted. The laboratory spikes were fortified with known concentrations of PFOA and 13CPFOA. The two sediment samples were extracted in one set. The set included one reagent blank and two reagent blanks fortified at known concentrations. For each sample, a laboratory duplicate of the sample and two laboratory matrix spikes were also extracted. The laboratory spikes were fortified with known concentrations PFOA and I3C PFOA. 8.0 RESULTS Analytical results for the analysis of PFOA in water samples are summarized in Table I. Analytical results for the analysis of PFOA in re-extracted water samples are summarized in Table 11. Analytical results for the analysis of PFOA in sludge samples are summarized in Table 111. Analytical results for the analysis of PFOA in sediment samples are summarized in Table IV. Quantitative results were obtained for all samples and analytes except for samples from sites DAL-GW-138R-0-060412,DAL-GW-138S-O060412, DAL-SD-OSPO1-0-0000, and DAL-SD-OSP02-0-0000 that are not reported (NR) due to quality control failures. Accuracies were assessed for each sample by reviewing the individual QC results obtained for each sample site. For the water samples, there were two laboratory and two or three field spike recovery results available for each sample site that were used to assess Exygen Research Page 22 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 the accuracy. In instances of failed laboratory or field spikes, recoveries associated with other spikes were used to assess sample accuracy. For the sludge and sediment samples, there were two laboratory spike recovery results available for each sample site that were used to assess the accuracy. In instances of failed laboratory spike recoveries, the samples were not reported due to the quality control failure. Fortification recoveries for PFOA in the water samples are detailed in Table V. The average percent recovery k standard deviation for PFOA in the water samples was 75 f 22%. Fortification recoveries for PFOA in the re-extracted water samples are detailed in Table VI. The average percent recovery & standard deviation for PFOA in the reextracted water samples was 99 f 16%. Fortification recoveries for PFOA in the sludge samples are detailed in Table VII. The average percent recovery for PFOA in the sludge sample was 69%. A standard deviation was not calculated for PFOA in sludge because only one sample was analyzed. Fortification recoveries for PFOA in the sediment samples are listed as NR in Table VIII. The average percent recovery f standard deviation for PFOA in the sediment samples was not calculated due to quality control failures. Fortification recoveries for I3CPFOA in the water samples are detailed in Table IX. The average percent recovery f standard deviation for 13CPFOA in the water samples was 75 f 26%. Fortification recoveries for I3C PFOA in the re-extracted water samples are detailed in Table X. The average percent recovery f standard deviation for 13CPFOA in the re-extracted water samples was 97 f 34%. Fortification recoveries for 13CPFOA in the sludge samples are detailed in Table XI. The average percent recovery f standard deviation for I3CPFOA in the sludge samples was 49 If: 16%. Fortification recoveries for 13CPFOA in the sediment samples are listed as NR in Table XII. The average percent recovery k standard deviation for 13CPFOA in the sediment samples was not calculated due to quality control failures. Percent recoveries have not been reported for samples that contained analyte levels three times or greater than the fortification level. Percent solids for sludge and sediment samples are detailed in Table XIII. 9.0 CONCLUSIONS Except as noted above, the ground water, sludge, and sediment samples were successhlly extracted and analyzed for PFOA according to analytical methods VOOOl780, VOOOl781, and V0001782 respectively. There were no circumstances that may have affected the data quality or integrity. Exygen Research Page 23 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 10.0 RETENTION OF DATA AND SAMPLES All original paper data generated by Exygen Research that pertains to this interim report will be shipped to the study director. This does not include facility-specificraw data such as instrument or temperature logs. Exact copies of all raw data, as well as a signed copy of the interim analytical report and all original facility-specific raw data, will be retained in the Exygen Research archives for the period of time specified in EPA TSCA Good Laboratory Practice Standards 40 CFR 792. Exygen Research Page 24 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 TABLES Exygen Research Page 25 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Table I. Summary of PFOA in Water Samples Exygen ID C0169333 C0169336 C0169336 Rep C0169337 C0169341 C0169341 Rep C0169342 C0169346 C0169346 Rep C0169347 C0169351 C0169351 Rep C0169352 C0169355 C0169355 Rep C0169356 C0169359 C0169359 Rep C0169360 C0169363 C0169363 Rep C0169364 C0169367 C0169367 Rep C0169368 C0170984 C0170984 Rep C0170985 coi 70988 CO170988 Rep C0170989 C0170992 CO170992 Rep C0170993 C0170996 C0170996 Rep C0170997 C0171000 C0171000 Rep C0171001 Client Sample ID DAL-GW-TRIPI-0-060411 DAL-GW-138L-0-060411 DAL-GW-138L-0-06041I * DAL-GW-138L-DB-060411 DAL-GW-138R-0-060412 DAL-GW-138R-0-060412' DAL-GW-138R-DB-060412 DAL-GW-1388-0-060412 DAL-GW-I38S-0-060412* DAL-GW-138s-DB-060412A DAL-GW-605R-0-060412 DAL-GW-605R-0-060412' DAL-GW-605R-DB-060412 DAL-GW-605L-0-060412 DAL-GW-605L-0-060412* DAL-GW-605L-DB-060412 DAL-GW-603s-0-060412 DAL-GW-603s-0-060412' DAL-GW-603s-DB-060412 DAL-GW-604L-0-060412 DAL-GW-604L-0-060412' DAL-GW-604L-DB-060412 DAL-GW-604s-0-060412 DAL-GW-604S-0-060412* DAL-GW-604s-DB-060412 DAL-GW-601R-0-060413 DAL-GW-601R-0-060413' DAL-GW-601R-DB-060413 DAL-GW-601S-0-060413 DAL-GW-601S-0-060413* DAL-GW-601S-DB-060413 DAL-GW-602R-0-060413 DAL-GW-602R-0-060413" DAL-GW-602R-DB-060413 DAL-GW-601L-0-060413 DAL-GW-601L-0-060413* DAL-GW-601L-DB-060413 DAL-GW-602s-0-060413 DAL-GW-602s-0-060413' DAL-GW-602s-0-060413 Analyte Found (ppt, nglL) C8 Acid PFOA Perfluorooctanoic Acid ND 21 9 239 238 NR NR NR NR NR NR 92.1 106 66.1 ND ND ND 317 32 1 392 1580 1490 1710 913 937 1070 NR NR NR NR NR NR 584 563 627 NR NR NR 557 523 563 Assessed Accuracy (+I- Yo) 30 50 50 50 NR NR NR NR NR NR 50 50 50 50 50 50 30 30 30 40 40 40 30 30 30 NR NR NR NR NR NR 30 30 30 NR NR NR 50 50 50 'Laboratory Duplicate A Sample was bottle was filled to 250 mL. ND = Not detected at or above the Limit of Quantitation (LOQ) of 25 nglL. NR = Not reported due to quality control failures, see Table IIfor re-extractiondata Exygen Research Page 26 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Table I. Summary of PFOA in Water Samples Continued Exygen ID CO 171004 C0171004 Rep C0171005 C0171008 C0171008 Rep C0171009 C0171012 C0171012 Rep C0171013 C0171016 C0171016 Rep C0171017 C0171020 C0171020 Rep C0171021 C0171025 C0171025 Rep C0171026 C0171030 C0171030 Rep C0171031 C0171035 C0171035 Rep C0171036 C0171039 C0171039 Rep C0171040 C0171043 C0171043 Rep C0171044 C0171047 C0171047 Rep C0171048 C0171051 C0171051 Rep C0171052 C0171055 C0171055 Rep C0171056 Client Sample ID DAL-GW-602L-0-060413 DAL-GW-602L-0-060413* DAL-GW-602L-DB-060413 DAL-GW-604R-0-060413 DAL-GW-604R-0-060413' DAL-GW-604R-DB-060413 DAL-GW-603R-0-060413 DAL-GW-603R-0-060413* DAL-GW-603R-DB-060413 DAL-GW-603L-0-060413 DAL-GW-603L-0-060413* DAL-GW-603L-DE060413 DAL-SW-BPPOI-0-060413 DAL-SW-BPP01-0-060413* DAL-SW-BPPOI-0-060413 DAL-SW-BPP02-0-060413 DAL-SW-BPP02-0-060413' DAL-SW-BPP02-DB-060413 DAL-SW-BPP03-0-060413 DAL-SW-BPP03-0-060413" DAL-SW-BPP03-DB-060413 DAL-SW-BCTOI-0-060412 DAL-SW-BCT01-0-060412' DAL-SW-BCT01-DB-060412 DAL-SW-BCT02-0-060412 DAL-SW-BCT02-0-060412' DAL-SW-BCT02-DB-060412 DAL-SW-BCT03-0-060412 DAL-SW-BCT03-0-0604 12' DAL-SW-BCTO3-DB-066412 DAL-SW-BC01-0-060412 DAL-SW-BCOI-0-060412' DAL-SW-BCOI-DB-060412 DAL-SW-BC02-0-060412 DAL-SW -BC02-0-060412* DAL-SW-BC02-DB-060412 DAL-SW-BC03-0-060414 DAL-SW-BC03-0-060414* DAL-SW-BC03-DB-060414 Analyte Found (ppt, nglL) C8 Acid PFOA Perfluorooctanoic Acid 2820 2940 3070 699 694 737 545 55 1 574 145 148 145 NR NR NR 248000 250000 249000 23 1000 248000 352000 NR NR NR 132000 126000 125000 24700 24700 28000 103 104 Ill 126 133 140 2370 2530 2660 Assessed Accuracy (%) 30 30 30 40 40 40 50 50 50 30 30 30 NR NR NR 30 30 30 30 30 30 NR NR NR 40 40 40 50 50 50 50 50 50 50 50 50 30 30 30 'Laboratory Duplicate ND = Not detected at or above the Limit of Quantitation (LOQ) of 25 ng/L. NR = Not reporteddue to quality controlfailures, see Table II for re-extractiondata Exygen Research Page 27 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Table I. Summary of PFOA in Water Samples Continued Exygen ID C0171059 C0171059 Rep C0171060 COI71063 C0171063 Rep C0171064 C0171067 CO171067 Rep C0171068 C0171071 C0171071 Rep C0171072 C0171075 C0171075 Rep C0171076 C0171079 C0171079 Rep C0171080 C0171084 C0171084 Rep C0171085 COI71088 C0171088 Rep C0171089 C0171092 C0171092 Rep C0171093 C0171096 C0171096 Rep C0171097 C0171100 COI 71 103 Client Sample ID DAL-SW-BC04-0-060414 DAL-SW-BC04-0-060414' DAL-SW-BC04-DB-060414 Analyte Found (ppt, nglL) C8 Acid PFOA Pemuorooctanoic Acid NR NR NR DAL-SW-BC05-0-060414 NR DAL-SW-BC05-0-060414' NR DAL-SW-BC05-DB-060414 NR DAL-SW-OSPOI-0-060413 335 DAL-SW-OSPOI-0-060413* 307 DAL-SW-OSP01-DB-060413 313 DAL-SW-OSP02-0-060413 NR DAL-SW-OSP02-0-060413' NR DAL-SW-OSP02-DB-060413 NR DAL-SW-OSPO3-0-060413 NR DAL-SW-OSP03-0-060413* NR DAL-SW-OSP03-DB-060413 NR DAL-GW-BPWELL-0-060413 NR DAL-GW-BPWELL-0-060413* NR DAL-GW-BPWELL-DB-060413 NR DPWS-WWI-DCTPOI-0-060413 DPWS-WWI-DCTPOI-0-060413' DPWS-WWI-DCTPOI-DB-060413 4200 4230 4390 DPWS-WWE-DCTPOI-0-060413" DPWS-WWE-DCTPOI-0-060413*" DPWS-WWE-DCTPOI-DB-060413 7030 7180 7020 DAL-WW - E FFO1-0-0604 13 DAL-WW-EFFOI-0-060413* DAL-WW-EFFOI-DB-060413 DAL-LCH-MCLFOI-0-060414 DAL-LCH-MCLFO1-0-060414* DAL-LCH-MCLFOI-DB-060414 4650 4460 4790 NR NR NR DAL-GW-TRIPI-0-060412 ND DAL-WW-TRIPI-0-060413 ND Assessed Accuracy (%) NR NR NR NR NR NR 40 40 40 NR NR NR NR NR NR NR NR NR 50 50 50 50 50 50 40 40 40 NR NR NR 30 30 'Laboratory Duplicate A Sample was bottle was filled to 230 mL. ND = Not detectedat or above the Limit of Quantitation(LOQ)of 25 ng/L. NR = Not reporteddue to quality controlfailures, see Table IIfor re-extractiondata Exygen Research Page 28 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Table 11. Summary of PFOA in Re-extracted Water Samples Exygen ID C0169341 C0169347 C0170984 C0170989 C0170997 C0171021 C0171035 C0171060 C0171063 C017 1071 C0171076 C0171079 C0171097 Client Sample ID DAL-GW-I 38R-0-060412 DAL-GW-138s-DB-060412 DAL-GW-601R-0-060413 DAL-GW-601S-DB-060413 DAL-GW -60 1L-D8-060413 DAL-SW-BPPOI-DB-060413 DAL-SW-BCTOI-0-060412 DAL-SW-BC04-DB-060414 DAL-SW-BC05-0-060414 DAL-SW-OSP02-0-060413 DAL-SW-OSP03-DB-060413 DAL-GW-BPW ELL-0-060413 DAL-LCH-MCLFO1-DB-060414 NR = Not reported due to quality control failures. Analyte Found (ppt, nglL) C8 Acid PFOA Pemuorooctanoic Acid NR NR 107 121 I9800 526000 29000 27700 782 2660 2650 64 1000 48700 Assessed Accuracy (+/- %) NR NR 30 30 30 30 30 30 30 30 30 30 30 Exygen Research Page 29 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Table 111. Summary of PFOA in Sludge Samples Exygen ID C0171106 C0171106 Rep Client Sample ID DPWS-SL-DCTPO1-0-0000 DPWS-SL-DCTP01-0-0000' Analyte Found (ppb, ndg) Dry Weight C8 Acid PFOA Perfluorooctanoic Acid 1860 1890 Assessed Accuracy (+I- %) 40 40 "Laboratory Duplicate Exygen Research Page 30 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Table IV. Summary of PFOA in Sediment Samples Exygen ID C0171107 C0171107 Rep C0171108 C0171108 Rep Client Sample ID DAL-SD-OSPOI-0-0000 DAL-SD-OSP01-0-0000' DAL-SD-OSP02-0-0000 DAL-SD-OSP02-0-0000* Analyte Found (ppb, nglg) C8 Acid PFOA Pemuorooctanoic Acid NR NR NR NR Assessed Accuracy (+I- %) NR NR NR NR 'Laboratory Duplicate NR = Not reported due to quality control failures. Exygen Research Page 31 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Table V. Matrix Spike Recovery of PFOA in Water Samples Sample Description DAL-GW-TRIP1 -LS-060411 (C0169334,l.O ppb Field Spike) DAL-GW-TRIP1-HS-060411 (COl60335. 10.0 ppb Field Spike) - Amount Spiked (nglL) Amt Found in Sample (nglL) 1000 ND 10000 ND C8 Acid PFOA Amount Recovered (nglL) 1040 10800 Recovery (%) 104 108 DAL-GW-138L-0-060411 (C0169336 Spk C, 10.0 ppb Lab Spike) 10000 219 DAL-GW-138L-0-060411 (C0169336 Spk D, 1000 ppb Lab Spike) 1000000 219 DAL-GW-138L-LS-060411 (C016933&10.0 ppb Field Spike) 10000 219 DAL-GW-138L-MS-060411 (C0169330.100 ppb Field Spike) 100000 219 DAL-GW-138L-HS-060411 (C0169340.1000 ppb Field Spike) 1000000 219 7990 78 896000 90 5760 55 38900 39 190000 19 DAL-GW-138R-0-060412 (CO169341Spk E, 10 ppb Lab Spike) 10000 NR DAL-GW-138R-0460412 (COl68341 Spk F,1000 ppb Lab Spike) 1000000 NR DAL-GW-138R-LS-060412 (CO169343.10 ppb Field Spike) 10000 NR DAL-GW-138R-MS-060412 (CO169344.100 ppb FieldSpike) 100000 NR DAL-GW-138R-HS-060412 (C0169345. 1000 ppb Field Spike) 1000000 NR NR NR NR NR NR NR NR NR NR NR DAL-GW-1388-0-060412 (C0169346 Spk C. 10 ppb Lab Spike) 10000 NR DAL-GW-1388-0-060412 (C0169346 Spk D. 1000 ppb Lab Spike) 1000000 NR DAL-GW-138s-LS-060412 (C0169346.10 ppb Field Spike) 10000 NR DAL-GW-138s-MS-060412 (CO169349.100 ppb Field Spike) 100000 NR DAL-GW-138s-HS-060412 (COl69350.1000 ppb Field Spike) 1000000 NR NR NR NR NR NR NR NR NR NR NR DAL-GW-605R-0-060412 (C0169351 Spk E. 0.1 ppb Lab Spike) DAL-GW-605R-0460412 (COl6935l Spk F. 1.0 ppb Lab Spike) DAL-GW-605R-LS-060412 (C0169353.0.1 ppb Field Spike) DAL-GW-605R-HS-060412 (C0169354,l.O ppb Field Spike)" 100 1000 100 800 92.1 92.1 92.1 92.1 175 83 833 74 138 46 566 59 DAL-GW-605L-0-060412 (C0169355 Spk G. 0.1 ppb Lab Spike) 100 ND DAL-GW-605L-0-060412 (C0160355 Spk H. 1.0 ppb Lab Spike) 1000 ND DAL-GW-605L-LS-060412 (C0169357.0.1ppb Field Spike) 100 ND DAL-GW-605L-HS-060412 (C0169356,l.O ppb FieldS p W 1000 ND 79.5 80 839 84 56.8 57 487 49 'Sample residue exceeds the spiking level significantly (3x spiking level); therefore, an accurate recovery value cannot be Calculated A Sample was bottle was filled to 250 mL instead of 200 mL, causing the spiking level to be modified. ND = Not defected at or above the Limit of Quantitation (LOCI) of 25 ngL. NR = Not reponeddue i o quality coniml failures, see Table VI for reextractiondata. Note: Since this summary table shows rounded results, rocoveryvaluer mayvarysllghtlyfrom the values In the raw data. Exygen Research Page 32 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Table V. Matrix Spike Recovery of PFOA in Water Samples Continued Sample Amount Spiked (nglL) Amt Found in Sample (nglL) C6 Acid PFOA Amount Recovered (nglL) Recovery (%) DAL-GW-603S-0-060412 (CO1693.59 Spk C. 0.1 ppb Lab Spike) 100 317 DAL-GW-603S-0-0604 12 (COl693.59 Spk D, 1.0 ppb Lab Splke) 1000 317 DAL-GW-603S-LS-060412 (C0169361.0.1 ppb Field Spike) 100 317 DAL-GW-603S-HS-060412 (C0169362,1.0ppb FieldSpike) 1000 317 418 1130 81 458 1040 72 DAL-GW-604L-0-060412 (COl69363 Spk E, 0.1 ppb Lab Spike) DAL-GW-604L-0-060412 (COW9363 Spk F. 1.0 ppb Lab Spike) DAL-GW-604L-LS-060412 (C0169365, 0.1 ppb Fleld Spike) DAL-GW-604L-HS-060412 (COl69366.1.0 ppb FleldSpike) 100 1000 100 1000 1580 1580 1580 1580 1680 2350 77 1670 2210 63 DAL-GW-604S-0-060412 (C0169367 Spk G. 0.1 ppb Lab Splke) 100 913 DAL-GW-604S-0-060412 (C0169367 Spk H, 1.0 ppb Lab Spike) 1000 913 DAL-GW-604S-LS-060412 (COl69369.0.1 ppb Field Spike) 100 913 DAL-GW-604S-HS-060412 (C0169370, 1.0 ppb Fleld Spike) 1000 913 987 1800 a9 1300 2040 113 DAL-GW-601R-0-060413 (C0170984 Spk C. 0.1 ppb Lab Spike) 100 NR DAL-GW-601R-0-060413 (C0170984 Spk D, 1.0 ppb Lab Spike) 1000 NR DALGW-6OlR-LS-060413 (C0170986. 0.1 ppb Field Spike) 100 NR DAL-GW-601 R-HS-060413 (C0170987.1.0 ppb Field Sp(ke) 1000 NR NR NR NR NR NR NR NR NR D A L - G W M 1S-0460413 (COl709SeSpk E. 0.1 ppb LabSpike) 100 NR DAL-GW-601 S-0-060413 (C01709.96Spk F. 1.0 ppb Lab Spike) 1000 NR DAL-GW-601S-LS-060413 (C017ooW. 0.1 ppb Field Spike) 100 NR DAL-GW-601 S-HS-060413 (C017wO1.1.0 ppb Field Spike) 1000 NR NR NR NR NR NR NR NR NR DAL-GW-602R-0-060413 (C0170992 Spk G. 0.1 ppb Lab Splke) 100 584 656 DAL-GW-602R-0-060413 (C017wO2 Spk H. 1.0 ppb Lab Spike) 1000 584 2010 143 DAL-GW-602R-LS-060413 (C0170994.0.1 ppb Field Spike) 100 584 762 DAL-GW-602R-HS-060413 (C017wO5.1.0 ppb Field Spike) - 1000 584 1880 130 'Sample residue exceeds the spiking level significanuy(3x spiking level); therefore, an accurate recoveryvalue cannot be calculated ND = Not detected at or abovethe Limit of bantitation (LOQ) of 25 ng/L. NR = Not reported due to quality mtrd failures, see Table VI for re-extractiondata. Note: Since thk summary table shows rounded resub, recovery values may vary slightly from the values In the raw data. Exygen Research Page 33 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.:PO000760 Table V. Matrix Spike Recovery of PFOA in Water Samples Continued Sample Description Amount Spiked (nglL) Amt Found in Sample C8 Acid PFOA Amount Recovered Recovery DAL-GW-601L-0-060413 (CO170996SpkC. 0.1 ppbLabSplke) 100 NR DAL-GW-601 L-0-060413 (C0170996Spk D. 1.0 ppbLabSpike) 1000 NR DAL-GW-601 L-LS-060413 (C0170998.0.1ppb Field Spike) 100 NR DAL-GW-601L-HS-060413 (C017OS99.1.0 ppb Field Spike) 1000 NR NR NR NR NR NR NR NR NR DAL-GW-602S-0-060413 (COl71000Spk E, 0.1 ppb LabSpike) 100 557 DAL-GW-602S-0-060413 (C0171MH)Spk F. 1.0 ppb LabSpike) 1000 557 DAL-GW-602S-LS-060413 (C0171WZ.0.1 ppb Fkld Spike.) 100 557 DAL-GW-602S-HS-060413 (C0171W3.1.0 ppb Field Spike) 1000 557 697 1280 72 573 1120 56 DAL-GW-602L-0-060413 (C0171004Spk G. 0.1 ppbLabSpike) DAL-GW-602L-0-060413 (C0171004Spk H. 1.0 ppbLabSpike) DAL-GW-602L-LS-060413 (C0171008.0.1 ppbFkidSpike) DAL-GW-602L-HS-060413 (C0171W7,1.0ppb Field Spike) 100 1000 100 1000 2820 2820 2820 2820 3250 3720 90 2870 4030 121 DAL-GW-604R-0-060413 (C0171006SpkC. 0.1 ppb LabSpike) 100 699 DAL-GW-604R-0-060413 (COl71008Spk 0.1.0 ppb LabSpike) 1000 699 DAL-GW-604R-LS-060413 (C0171010.0.1 ppb Field Spike) 100 699 DAL-GW-604R-HS-060413 (CO171011.1.0 ppbField Spike) 1000 699 DAL-GW-603R-0-060413 (C0171012Spk E. 0.1 ppb LabSpike) 100 545 DAL-GW803R-0-060413 (C0171012Spk F. 1.0 ppb LabSpike) 1000 545 DAL-GW-603R-LS-060413 (C0171014.0.1 ppb Field Spike) 100 545 DAL-GW-603R-HS-060413 (C0171015.1.0 ppb Field Spike) 1000 545 780 1440 74 789 1340 64 623 1230 69 682 1100 56 DAL-GWg03L-0-060413 (C0171016Spk G, 0.1ppb LabSpike) 100 145 DAL-GW-603L-0-060413 (C0171016Spk H. 1.0 ppbLabSpike) 1000 145 DAL-GW-603L-LS-OM)413 (C0171018.0.1 ppb Field Spike) 100 145 DAL-GW-603L-HS-060413 ~C0171019.1.0ppb Field Spike) 1000 145 249 104 1050 91 214 69 882 74 p p 'Sample residue exceeds the spiking level significantly (3x spiking level); therefore, an accurate recovely value cannot be calculated ND = Not detected at or above the Limit of Cuantitation( L a ) of 25 ng/L. NR = Not reporleddue to qualitycmtrol failures. see Table VI for re-extmctim data. Note: Slnce this summary table shows rounded result.. recovery values may vary slightly from the values in the raw data. Exygen Research Page 34 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Table V. Matrix Spike Recovery of PFOA in Water Samples Continued Sample Description DAL-SW-BPPO1-0-060413 (C0171020 Spk C, 1.0 ppb Lab Spike) DAL-SW-BPPO1-0-060413 (COl71020 Spk 0 , l O ppb Lab Spike) DAL-SW-BPPO1-LS-060413 (C0171022.1.0 ppb Fkid Splke) DAL-SW-BPWI-MS-060413 (C0171023.10 ppb FIeld Spike) DAL-SW-BPPO1-HS-060413 (C0171024,100 ppb Field Spike) DAL-SW-BPW2-0-060413 (C0171025 Spk E. 1.0 ppb Lab Spike) DAL-SW-BPP02-0-060413 (C0171025 Spk F, 100 ppb Lab Spike) DAL-SW-BPP02-LS-060413 (C0171027.1.0 ppb Field Splke) DAL-SW-BPW2-MS-060413 (C0171028.10 ppb Fleld Spike) DALSW-BPP02-HS-060413 (C0171029.100 ppb Field Spike) DAL-SW-BPP03-0-060413 (C01710M Spk G. 1.0 ppb Lab Spike) DAL-SW-BPP03-0-060413 (C0171030 Spk H. 100 ppb Lab Spike) DAL-SW-BPP03-LS-060413 (C0171032.1.0 ppb Field Spike) DAL-SW-BPP03-MS-060413 (C0171033.10 ppb Fleld Spike) DAL-SW-BPP03-HS-060413 (C0171034.100 ppb Fleid Spike) DALSW-BCT01-0-060412 (C0171035 Spk C, 1.0 ppb Lab Spike) DAL-SW-BCT01-0-060412 (C017i035Spk D. i o ppb Lab Spik.) DAL-SW-BCTO1-LS-060412 (C0171037.1.0 ppb Field Spike) DAL-SW-BCTOI -HS-060412 (C01710Js.10 ppb Fleld Spike) DAL-SW-BCT02-0-060412 (C0171039 Spk E. 1.0 ppb Lab Spike) DAL-SW-BCT02-0-060412 (COl71039 Spk F. I O ppb Lab Spike) DALSW-BCT02-LS-060412 (C0171041.1.0 ppb Field Spike) DAL-SW-BCT02-HS-060412 (C0171042.10 ppb Fleld Spike) Amount Spiked (nglL) 1000 100000 1000 10000 100000 1000 100000 1000 10000 100000 1000 100000 1000 10000 100000 1000 10000 1000 10000 1000 10000 1000 10000 Amt Found in Sample (nglL) NR NR NR NR NR 248000 248000 248000 248000 248000 231000 231000 231000 231000 231000 NR NR NR NR 132000 132000 132000 132000 C8 Acid PFOA Amount Recovered (nglL) NR NR NR NR NR 224000 319000 226000 219000 324000 229000 352000 231000 206000 346000 NR NR NR NR 144000 138000 119000 138000 Recovety (%) NR NR NR NR NR 71 76 121 115 NR NR NR NR 60 60 'Sample residue exceeds the spiking level significantly (3x spiking level), therefore,an accurate recovery value cannot be calculated ND = Not detected at or above the Limit of Quantitation (LOQ) of 25 ngR NR = Not reported due to quality control failures. see Table Vi for re-extraction data Note: Since this summary table shows rounded results. recoveryvalues may vary slightly from the values I n the raw data. Exygen Research Page 35 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Table V. Matrix Spike Recovery of PFOA in Water Samples Continued Sample - Amount Spiked Amt Found in Sample C8 Acid PFOA Amount Recovered Recovery DALSW-BCT03-0-060412 (C0171043 Spk G. 1.0 ppb Lab Spike) DALSW-BCT03-0-060412 (C0171043 Spk H, 10 ppb Lab Spike) DAL-SW-BCT03-LS-0604 12 (C0171045.1.0 ppb Fleid Spike) DAL-SW-BCT03-HS-060412 (C0171M6.10 ppb Field Spike) DALSW-BCO1-0-060412 (C0171047 Spk C. 1.0 ppb Lab Spike) DAL-SW-BCO1-0-060412 (C0171M7 Spk D. I O ppb Lab Spike) DAL-SW-BC01-LS-060412 (C0171049.1.0 ppb Field Spike) DAL-SW-BCO1-HS-060412 (C01710M. 10 ppb FieldSpike) DAL-SW-BC02-0-060412 (C0171051 Spk E. 1.0 ppb Lab Spike) DALSW-BC02-0-060412 (C0171051 Spk F. 10 ppb Lab Spike) DALSW-BC02-LS-060412 (C0171053.1.0 ppb Field Spike) DAL-SW-BC02-HS-060412 (C0171054.10 ppb Field Splke) DAL-SW-BC03-0-060414 (C0171055 SpkG, 1.0 ppb Lab Spike) DAL-SW-BC030-060414 (C0171055 Spk H, 10 ppb Lab Spike) DAL-SW-BC05LS-060414 (C0171057.1.0 ppb Field Spike) DAL-SW-BC03-HS-060414 (C01710M. 10 ppb Field Spike) DAL-SW-BC04-0-0604 14 (C0171059 Spk C, 1.0 ppb Lab Spike) DAL-SW-BC04-0-060414 (C0171059Spk D, 10 ppbLab Spike) DAL-SW-BC04-LS-060414 (C0171061.1.0 ppb Fleid Spike) DAL-SW-BC04-HS-0604 14 (C0171062.10 ppb Field Spike) DAL-SW-BC05-0-060414 (C0171063 Spk E, 1.0 ppb Lab Spike) DAL-SW-BC05-0-060414 (C0171063Spk F. I O ppbLabSpike) DAL-SW-BC05LS-060414 (C0171065.1.0 ppb Fleld Spike) DAL-SW-BC05-HS-060414 (C0171066,10 ppb Field Spike) 1000 10000 1000 10000 1000 10000 1000 10000 1000 10000 1000 10000 1000 10000 1000 10000 1000 10000 1000 10000 1000 10000 1000 10000 24700 24700 24700 24700 103 103 103 103 126 126 126 126 2370 2370 2370 2370 NR NR NR NR NR NR NR NR 26000 32100 74 26500 30400 57 863 76 7760 77 647 54 4780 47 850 72 8090 80 654 53 5500 54 3360 99 9900 75 3300 93 9200 68 NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR `Sample residue exceeds the spiking level significantly (3x spiking level); therefore, an accurate recovery value cannot be calculated ND = Not detected a1or above the Limit of Quantitation (LOQ) of 25 ng/L. NR = Not reporteddue to quality controlfailures. see Table VI for re-extraction data Note:Slnce this summary table shows rounded results, recovery values may vary slightly from the values In the raw data. Exygen Research Page 36 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Table V. Matrix Spike Recovery of PFOA in Water Samples Continued Sample Description DALSW-OSPOl-0-060413 (CO171W7 Spk G. 0.1 ppb Lab Spike) DAL-SW-OSPOI-0-060413 (CO171W7SpkH.1.OppbLlbSpike) DAL-SW-OSPOI-LS-060413 (CO171OW, 0.1 ppb Field Splke) DAL-SW-OSPOI-HS-060413 (C0171070,l.O ppb Field Spike) DAL-SW-OSWZ-0-060413 (CO171071Spk C. 1.0 ppb Lab Spike) DAL-SW-OSP02-0-060413 (C0171071Spk D. 10 ppb Llb Spike) DAL-SW-OSPOZ-LS-060413 (CO171073,l.O ppb Fleld Spike) DAL-SW-OSP02-HS-060413 (C0171074,10ppb Fhld Spike) DAL-SW-OSP03-0-060413 (C0171075 Spk E, 1.0 ppb Lab Spike) OAL-SW-OSP03-0-060413 (CO171075 Spk F. 10 ppb Lab Spike) DAL-SW-OSP03-LS-060413 (CO171077.1.0 ppb Field Spike) DAL-SW-OSP03-HS-060413 (C0171078.10 ppb Field Spike) Amount Spiked (ne/L) 100 1000 100 1000 100 1000 100 1000 100 1000 100 1000 Amt Found in Sample (nglL) 335 335 335 335 NR NR NR NR NR NR NR NR C8 Acld PFOA Amount Recovered (nglL) 375 1070 402 97 1 NR NR NR NR NR NR NR NR Recovery (X) 74 64 NR NR NR NR NR NR NR NR DAL-GW-BPWELL-0-060413 (CO17107BSpkG,10ppbLabSplke) DAL-GW-BPWELL-0-060413 (CO171079Spk H. IO00 ppb Lab Spike) DAL-GW-BPWELL-LS-060413 (CO171001.10 ppb Field Splke) DAL-GW-BPWELL-MS-060413 (COl7lM2,100 ppb Fleld Spike) DAL-GW-BPWELL-HS-060413 (CO171003. IWO ppb Field Spike) DPWS-WWI-DCTWI -0-060413 (CO17iOM Spk C. 10 ppb Llb Splke) DPWS-WWI-DCTP01-0-060413 (COITIOMSpk D. 100 ppb Lab Splke) DPWS-WW I-DCTP01-LS-060413 (CO1710M,lO ppb Fkid Spike) DPWS-WWI-DCTPO1-HS-060413 (CO171M7,100ppb Field Spike) DPWS-WWE-DCTPOI-0-060413 (CM71OMSpk E, 10 ppb Lab Splker DPWS-WWE-OCTPOI-0-060413 (CO171OM Spk F. 100 ppb Lab Spike)' DPWS-WWE-DCTPOI -LS-060413 (C0171OW. 10 ppb Field Splke) DPWS-WWE-DCTPOI-HS-060413 (CO171W1,1W ppb fleid Spike) 10000 I000000 10000 100000 1000000 10000 100000 10000 100000 10000 100000 10000 100000 NR NR NR NR NR 4200 4200 4200 4200 7030 7030 7030 7030 NR NR NR NR NR NR NR NR NR NR 13100 89 96500 92 9220 50 46700 43 11700 47 95100 88 12600 56 64300 57 'Sample residue exceeds the spiking level SlgnilicanUy (3x spiking level); therefore,an accurate recovery MlUe cannot be calculated A Samplewas bonkwas filled to 230 mL insteadof2M) mL. ND = Not detected at or above the Limitof Quantitation (LOQ)of25 nglL. NR = Not reporteddue to qualitycontrolfailures, see Table VI for reextractiondata Note: Sincethissummarytable shows roundedresults,recoveryvalues mayvaryslightlyfrom the values In the raw data. Exygen Research Page 37 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Table V. Matrix Spike Recovery of PFOA in Water Samples Continued Sample Description - Amount Spiked (nglL) Amt Found in Sample (nglL) C8 Acid PFOA Amount Recovered (nglL) - Recovery (x) DAL-WW-EFF01-0-060413 (C0171092 Spk G. 1.0 ppb Lab Spike) DAL-WW-EFFO1-0-060413 (C0171092 Spk H, 10 ppb Lab Spike) DAL-WW-EFFO1-LS-060413 (C0171094.1.0 ppb Field Spike) DAL-WW-EFF01-HS-060413 (C0171095.10 ppb Field Spike) DAL-LCH-MCLFO1-0-0604 14 (C0171096 Spk C. 10 ppb Lab Spike) DAL-LCH-MCLFO 1-0-0604 14 (C0171096 Spk D. 100 ppb Lab Spike) DAL-LCH-MCLFO1-LS-060414 (C0171098.10 ppb Field Spike) DAL-LCH-MCLFO1-HS-060414 (C0171099.1W ppb Field Spike) DAL-GW-TRIP1-LS-060412 (C0171101.1.0 ppb Field Spike) DALGW-TRIP1 -HS-060412 (C0171102. I O ppb Field Spike) DAL-WW-TRIP1-LS-060413 (C0171104.1.0 ppb Field Spike) DAL-WW-TRIP1-HS-060413 (C0171105.10 ppb Fleld Spike) 1000 10000 1000 10000 10000 100000 10000 100000 1000 10000 1000 10000 4650 4650 4650 4650 NR NR NR NR ND ND ND ND 5250 11700 71 5890 11200 66 NR NR NR NR NR NR NR NR 776 78 7950 80 857 86 9500 95 Average: IS Standard Deviation: 22 'Sample residue exceeds the spiking level significantly (3x spiking level); therefore. an accurate recovery value cannot be calculated ND = Not detected at or above the Limit of Quantilation (LOQ)of 25 nglL. NR = Not reported due to quality control failures, see Table VI for reextraclion data. Note: Since this summary table ahows rounded results. recovery values may vary slightly from the values in the raw data. Exygen Research Page 38 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Table VI. Matrix Spike Recovery PFOA in Re-extracted Water Samples Sample Description DAL-GW-138R-HS-060412 (COl80345. 1000 ppbFieldSpike) DAL-GW-138s-DB-060412 (C0189347 Spk E. 100 ppb Lab Splke) DAL-GW-138s-MS-060412 (COl8#34#.100 ppb FieldSplke) DAL-GW-601R-LS-060413 (C0170086.0.1 ppbFieldSplke) DAL-GW-601S-DB-060413 (C0170089 SpkG, 0.1 ppbLabSpike) DAL-GW-601S-LS-060413 (C0170990.0.1 ppb FieldSplke) DAL-GW-601L-DB-060413 (C0170997 Spk E, 10 ppb Lab Spike) DAL-SW-BPPO1-DB-060413 (C0171021Spk E. 1000 ppbLabSpike) DAL-SW-BCTO1-HS-060412 (C0171038.10 ppb FieldSpike) DALSW-BC04-HS-060414 (C0171082.10 ppb FieldSplke) DAL-SW-BC05-LS-060414 (C0171065,l.O ppb Field Spike) DAL-SW-OSP02-HS-060413 (C0171074.10 ppb FieldSplke) DAL-SW-OSP03-DB-060413 (C0171076 Spk G, 1.0 ppb Lab Spike) DAL-GW-BPWELL-HS-060413 (C0171083,lo00 ppbFieldSpike) DAL-LCH-MCLFO1-HS-060414 (COl71099.100 ppb Fleid Splke) Amount Spiked (nglL) 1000000 100000 100000 100 100 100 10000 1000000 10000 10000 1000 1000 1000 1000000 100000 Amt Found in Sample (nglL) C8 Acid PFOA Amount Recovered (nglL) NR NR NR NR NR NR 107 222 121 223 121 220 19800 30100 526000 1790000 29000 37300 27700 39700 782 1520 2660 3610 2650 641000 3620 1540000 48700 132000 Recovery (%) NR NR NR 115 102 99 103 126 83 120 74 95 97 90 83 Average: 99 Standard Deviation: 16 'Sample residueexceedsthe spiking levelsignificantly (3x spiking level);therefore, an accuraterecoveryvaluecannotbecalculated NR = Notreporteddueto quality controlfailures. Note: Sincethis summarytable shows roundedresults. recoveryvalues mayvary slightly from thevalues in the raw data. Exygen Research Page 39 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Table VII. Matrix Spike Recovery of PFOA in Sludge Samples Sample Description Amount Spiked (ng/g) Amt Found in Sample (ng/g) wet weight Amount Recovered (ng/g) wet weight Recovery (%) DPWS-SL-DCTPOI-0-0004 (C0171106 Spk C, 4 ppb Spike) 4 424 DPWS-SL-DCTPO1-0-0000 (C0171106 Spk D, 400 ppb Spike) 400 424 I I 439 70 1 69 Average: 69 Standard Deviation: NA 'Sample residue exceeds the spiking levelsignificantly (3x spiking level);therefore,an accurate recoveryvalue cannot be calculated. Note: Since this summarytable shows rounded results, recoveryvalues may vary slightlyfrom the values in the raw data. Exygen Research Page 40 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.:PO000760 Table VIII. Matrix Spike Recovery PFOA in Sediment Samples Sample Description DAL-SD-OSP01-0-0000 (C0171107 Spk C, 4 ppb Lab Spike) DAL-SD-OSPOl-0-0000 (C0171107Spk D, 400 ppb Lab Spike) DAL-SD-OSP02-0-0000 (C0171108Spk E,4 ppb Lab Spike) DAL-SD-OSP02-0-0000 (C0171108 Spk F, 400 ppb Lab Spike) Amount Spiked (ng/g) Amt Found in Sample Jng/g) wet weight C8 Acid PFOA Amount Recovered (ng/g) wet weight 4 NR NR 400 NR NR 4 NR NR 400 NR NR NR = Not reporteddue to quality controlfailures. Recovery (%) NR NR NR NR Exygen Research Page 41 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Table IX. Surrogate Spike Recovery of 13CPFOA in Water Samples fiygen ID C0169333 C0169334 C0169335 C0169336 Spk C C0169336 Spk D C0169336 C0169336 Rep C0169337 C0169338 C0169339 C0169340 C0169341 Spk E C0169341 Spk F C0169341 C0169341 Rep C0169342 C0169343 C0169344 C0169345 CO169346 Spk C C0169346 Spk D C0169346 C0169346 Rep C0169347 C0169348 C0169349 C0169350 C0169351 Spk E C0169351 Spk F C0169351 CO169351 Rep C0169352 C0169353 C0169354 C0169355 Spk G C0169355 Spk H C0169355 C0169355 Rep C0169356 C0169357 C0169358 Sample Description DAL-GW-TRIP1-0-060411 DAL-GW-TRIP1-LS-060411 DAL-GW-TRIP1 -HS-060411 DAL-GW-138L-0-060411 DAL-GW-138L-0-060411 DAL-GW-138L-0-060411 DAL-GW-138L-0-060411 DAL-GW-138L-DB-060411 DAL-GW-138L-LS-060411 DAL-GW-138L-MS-060411 DAL-GW-138L-HS-060411 DAL-GW-138R-0-060412 DAL-GW-138R-0-060412 DAL-GW-138R-0-060412 DAL-GW-138R-0-060412 DAL-GW-138R-DB-060412 DAL-GW-138R-LS-060412 DAL-GW-138R-MS-060412 DAL-GW-138R-HS-060412 DAL-GW-138s-0-060412 DAL-GW-1388-0-060412 DAL-GW-1388-0-060412 DAL-GW-1388-0-060412 DAL-GW-138s-DB-060412A DAL-GW-138s-LS-060412 DAL-GW-138s-MS-060412 DAL-GW-138s-HS-060412 DAL-GW-60513-0-060412 DAL-GW-605R-0-060412 DAL-GW-605R-0-060412 DAL-GW-605R-0-060412 DAL-GW-605R-DB-060412 DAL-GW-605R-LS-060412 DAL-GW-605R-HS-060412" DAL-GW-605L-0-060412 DAL-GW-605L-0-060412 DAL-GW-605L-0-060412 DAL-GW-605L-0-060412 DAL-GW-605L-DB-060412 DAL-GW-605L-LS-060412 DAL-GW-605L-HS-060412 Amount Spiked (nglL) I'C-PFOA Amount Recovered (nW Recovery (%) 500 546 109 1000 1070 107 10000 10700 107 10500 8240 78 1000500 861000 86 500 41 3 83 500 450 90 500 466 93 10000 5020 50 100000 37000 37 1000000 166000 17 10500 NR NR 1000500 NR NR 500 NR NR 500 NR NR 500 NR NR 10000 NR NR 100000 NR NR 1000000 NR NR 10500 NR NR 1000500 NR NR 500 NR NR 500 NR NR 400 NR NR 10000 NR NR 100000 NR NR 1000000 NR NR 600 478 80 1500 1090 73 500 371 74 500 401 80 500 393 79 100 53.8 54 800 440 55 600 546 91 1500 1230 82 500 403 81 500 438 88 500 512 102 100 54.2 54 1000 449 45 A Sample was bottle was filled to 250 mL, causing the IJCPFOA spiking level to be modified. NR = Not reported due to quality control failures. see Table X for re-extraction data. Note: Since this summary table shows rounded results, recoveryvalues may vary slightly from the values In the raw data. Exygen Research Page 42 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Table IX. Surrogate Spike Recovery of 13CPFOA in Water Samples Continued Exygen ID C0169359 Spk C C0169359 Spk D C0169359 C0169359 Rep CO169360 C0169361 CO169362 C0169363 Spk E C0169363 Spk F C0169363 CO169363 Rep C0169364 C0169365 C0169366 C0169367 Spk G C0169367 Spk H C0169367 C0169367 Rep C0169368 C0169369 C0169370 C0170984 Spk C C0170984 Spk D COI 70984 C0170984 Rep COI 70985 CO170986 C0170987 C0170988 Spk E C0170988 Spk F C0170988 C0170988 Rep C0170989 C0170990 C0170991 C0170992 Spk G C0170992 Spk H C0170992 C0170992 Rep C0170993 C0170994 C0170995 Sample Description DAL-GW-603s-0-060412 DAL-GW-603s-0-060412 DAL-GW-603s-0-060412 DAL-GW-603s-0-060412 DAL-GW-603s-DB-060412 DAL-GW-603s-LS-060412 DAL-GW-603s-HS-060412 DAL-GW-604L-0-060412 DAL-GW-604L-0-060412 DAL-GW-604L-0-060412 DAL-GW-604L-0-060412 DAL-GW-604L-DB-060412 DAL-GW-604L-LS-060412 DAL-GW-604L-HS-060412 DAL-GW-604s-0-060412 DAL-GW-604s-0-060412 DAL-GW-604s-0-060412 DAL-GW-604s-0-060412 DAL-GW-604s-DB-060412 DAL-GW-604s-LS-060412 DAL-GW-604s-HS-060412 DAL-GW-601R-0-060413 DAL-GW-601 R-0-060413 DAL-GW-601R-0-060413 DAL-GW-601R-0-060413 DAL-GW-601R-DB-060413 DAL-GWdOl R-LS-060413 DAL-GW-601R-HS-060413 DAL-GWdOl S-0-060413 DAL-GW-601S-0-060413 DAL-GWSOl S-0-060413 DAL-GW-601S-0-060413 DAL-GW-601S-DB-060413 DAL-GW-601S-LS-060413 DAL-GW-601S-HS-060413 DAL-GW-602R-0-060413 DAL-GW-602R-0-060413 DAL-GW-602R-0-060413 DAL-GW-602R-0-060413 DAL-GW-602R-DB-060413 DAL-GW-602R-LS-060413 DAL-GW-602R-HS-060413 Amount Spiked (ngW 600 1500 500 500 500 100 1000 600 1500 500 500 500 100 1000 600 1500 500 500 500 100 1000 600 1500 500 500 500 100 1000 600 1500 500 500 500 100 1000 600 1500 500 500 500 100 1000 "c-PFOA Amount Recovered (ngW Recovery (%) 520 87 1220 81 395 79 41 1 82 51 1 102 72.9 73 656 66 547 91 1350 90 431 86 440 88 446 89 62.7 63 536 54 491 82 1270 85 430 86 420 84 472 94 66.2 66 538 54 NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR 530 88 2280 152 974 195 948 190 94 1 188 120 120 1050 105 NR = Not reported due to qualitycontrol failures, see Table X for re-extractiondata. Note: Since this summatytable shows rounded results, recoveryvalues may vary slightly from the values in the raw data. Exygen Research Page 43 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Table IX. Surrogate Spike Recovery of 13CPFOA in Water Samples Continued Exyeen ID C0170996 Spk C C0170996 Spk D C0170996 C0170996 Rep C0170997 C0170998 C0170999 C0171000 Spk E C0171000 Spk F C0171000 COI71000 Rep C0171001 C0171002 C0171003 C0171004 Spk G C0171004 Spk H C0171004 C0171004 Rep C0171005 C0171006 C0171007 C0171008 Spk C C0171008 Spk D C0171008 C0171008 Rep C0171009 C0171010 C0171011 C0171012 Spk E COi71012 Spk F C0171012 C0171012 Rep C0171013 C0171014 C0171015 C0171016 Spk G C0171016 Spk H C0171016 C0171016 Rep C0171017 C0171018 C0171019 Sample Description DAL-GW-601L-0-060413 DAL-GW-601L-0-060413 DAL-GW-601L-0-060413 DAL-GW-601L-0-060413 DAL-GW-601 L-DB-060413 DAL-GW-601L-LS-060413 DAL-GW-601 L-HS-060413 DAL-GW-602S-0-060413 DAL-GW-602s-0-060413 DAL-GW-602S-0-060413 DAL-GW-602s-0-060413 DAL-GW-602s-DB-060413 DAL-GW-602s-LS-060413 DAL-GW-602s-HS-060413 DAL-GW-602L-0-060413 DAL-GW-602L-0-060413 DAL-GW-602L-0-060413 DAL-GW-602L-0-060413 DAL-GW-602L-DB-060413 DAL-GW-602L-LS-060413 DAL-GW-602L-HS-060413 DAL-GW-604R-0-060413 DAL-GW-604R-0-060413 DAL-GW-604R-0-060413 DAL-GW-604R-0-060413 DAL-GW-604R-DB-060413 DAL-GW-604R-LS-060413 DAL-GW-604R-HS-060413 DAL-GW-603R-0-060413 DAL-GW-603R-0-060413 DAL-GW-603R-0-060413 DAL-GW-603R-0-060413 DAL-GW-603R-DB-060413 DAL-GW-603R-LS-060413 DAL-GW-603R-HS-060413 DAL-GW-603L-0-060413 DAL-GW-603L-0-060413 DAL-GW-603L-0-060413 DAL-GW-603L-0-060413 DAL-GW-603L-DB-060413 DAL-GW-603L-LS-060413 DAL-GW-603L-HS-060413 ls~-~~~~ Amount Spiked (nglL) Amount Recovered (ngW Recovery (%) 600 NR NR 1500 NR NR 500 NR NR 500 NR NR 500 NR NR 100 NR NR 1000 NR NR 600 598 100 1500 1210 81 500 473 95 500 449 90 500 469 94 100 53.9 54 1000 581 58 600 557 93 1500 1330 89 500 423 85 500 437 87 500 448 90 100 67.6 68 1000 710 71 600 508 85 1500 1160 77 500 433 87 500 428 86 500 448 90 100 68.3 68 1000 575 58 600 541 90 1500 1140 76 500 436 87 500 433 a7 500 445 89 100 68.6 69 1000 486 49 600 555 93 1500 1350 90 500 434 87 500 438 88 500 439 88 100 76.6 77 1000 676 68 NR = Not reported due to quality control failures, see Table X for re-extraction data. Note: Since this summary table shows rounded results, recoveryvalues may vary slightly from the values in the raw data. Exygen Research Page 44 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Table IX. Surrogate Spike Recovery of 13CPFOA in Water Samples Continued Wgen ID C0171020 Spk C C0171020 Spk D C0171020 C0171020 Rep C0171021 C0171022 C0171023 C0171024 C0171025 Spk E C0171025 Spk F C0171025 C0171025 Rep C0171026 C0171027 C0171028 C0171029 C0171030 Spk G C0171030 Spk H C0171030 C0171030 Rep C0171031 C0171032 C0171033 C0171034 C0171035 Spk C C0171035 Spk D C0171035 C0171035 Rep C0171036 C0171037 C0171038 C0171039 Spk E CO171039 Spk F C0171039 C0171039 Rep C0171040 C0171041 C0171042 C0171043 Spk G C0171043 Spk H C0171043 C0171043 Rep C0171044 C0171045 C0171046 Sample Description DAL-SW-BPPO1-0-060413 DAL-SW-BPPO1-0-060413 DAL-SW-BPPO1-0-060413 DAL-SW-BPPO1-0-060413 DAL-SW-BPPO1-DB-060413 DAL-SW-BPPOI-LS-060413 DAL-SW-BPPO1-MS-060413 DAL-SW-BPPO1-HS-060413 DAL-SW-BPPO2-0460413 DAL-SW-BPP02-0-060413 DAL-SW-BPPO2-0460413 DAL-SW-BPP02-0-060413 DAL-SW-BPP02-DB-060413 DAL-SW-BPP02-LS-060413 DAL-SW-BPP02-MS-060413 DAL-SW-BPPO2-HS-060413 DAL-SW-BPP03-0-060413 DAL-SW-BPP03-0-060413 DAL-SW-BPP03-0-060413 DAL-SW-BPP03-0-060413 DAL-SW-BPP03-DB-060413 DAL-SW-BPP03-LS-060413 DAL-SW-BPP03-MS-060413 DAL-SW-BPP03-HS-060413 DAL-SW-BCTO1-0-060412 DAL-SW-BCTO1-0-060412 DAL-SW-BCTO1-0-060412 DAL-SW-BCTO1-0-060412 DAL-SW-BCTO1-DB-060412 DAL-SW-BCTO1-LS-060412 DAL-SW-BCT01-HS-060412 DAL-SW-BCT02-0-060412 DAL-SW-BCT02-0-060412 DAL-SW-BCT02-0-060412 DAL-SW-BCT02-0-060412 DAL-SW-BCT02-DB-060412 DAL-SW-BCT02-LS-060412 DAL-SW-BCT02-HS-060412 DAL-SW-BCT03-0-060412 DAL-SW-BCT03-0-060412 DAL-SW-BCT03-0-060412 DAL-SW-BCT03-0-060412 DAL-SW-BCT03-DB-060412 DAL-SW-BCTO3-LS-060412 DAL-SW-BCT03-HS-060412 Amount Spiked (ng/L) 1500 100500 500 500 500 1500 10500 100500 1500 100500 500 500 500 1500 10500 100500 1500 100500 500 500 500 1500 10500 100500 1500 10500 500 500 500 1000 10000 1500 10500 500 500 500 1000 10000 1500 10500 500 500 500 lo00 10000 'c-PFOA Amount Recovered (nW Recovery (%) NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR 770 51 84400 84 146 29 134 27 134 27 366 24 3820 36 45000 45 81 1 54 86100 86 152 30 144 29 138 28 404 27 4230 40 59800 60 NR NR NR NR NR NR NR NR NR NR NR NR NR NR 935 62 7620 73 152 30 147 29 143 29 379 38 4980 50 1110 74 7610 72 276 55 266 53 267 53 585 59 5130 51 NR = Not reported due to quality control failures, see Table X for re-extraction data. Note: Since this summary table shows rounded results, recoveryvalues may vary slightly from the values In the raw data. Exygen Research Page 45 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Table IX. Surrogate Spike Recovery of 13CPFOA in Water Samples Continued Exygen ID C0171047 Spk C C0171047 Spk D C0171047 C0171047 Rep C0171048 C0171049 C0171050 C0171051 Spk E C0171051 Spk F C0171051 C0171051 Rep C0171052 C0171053 C0171054 COI71055 Spk G C0171055 Spk H C0171055 C0171055 Rep C0171056 C0171057 C0171058 C0171059 Spk C C0171059 Spk D C0171059 C0171059 Rep C0171060 C0171061 C0171062 C0171063 Spk E C0171063 Spk F C0171063 C0171063 Rep C0171064 C0171065 C0171066 C0171067 Spk G C0171067 Spk H C0171067 COI71067 Rep C0171068 C0171069 CO171070 Sample Description DAL-SW-BC01-0-060412 DAL-SW-BCO1-0-060412 DAL-SW-BC01-0-060412 DAL-SW-BCOI-0-060412 DAL-SW-BCO1-DB-060412 DAL-SW-BCOI -LS-060412 DAL-SW-BC01-HS-060412 DAL-SW-BC02-0-060412 DAL-SW-BC02-0-060412 DAL-SW-BC02-0-060412 DAL-SW-BC02-0-060412 DAL-SW-BC02-DB-060412 DAL-SW-BC02-LS-060412 DAL-SW-BC02-HS-060412 DAL-SW-BC03-0-060414 DAL-SW-BC03-0-060414 DAL-SW-BC03-0-060414 DAL-SW-BC03-0-060414 DAL-SW-BC03-DB-060414 DAL-SW-BC03-LS-060414 DAL-SW-BC03-HS-060414 DAL-SW-BCO4-0-060414 DAL-SW-BC04-0-060414 DAL-SW-BC04-0-060414 DAL-SW-BC04-0-060414 DAL-SW-BC04-DB-060414 DAL-SW-BC04-LS-060414 DAL-SW-BC04-HS-060414 DAL-SW-BC05-0-060414 DAL-SW-BC05-0-060414 DAL-SW-BC05-0-060414 DAL-SW-BC05-0-060414 DAL-SW-BC05-DB-060414 DAL-SW-BC05-LS-060414 DAL-SW-BC05-HS-060414 DAL-SW-OSPO1-0-060413 DAL-SW-OSPO1-0-060413 DAL-SW-OSPO1-0-060413 DAL-SW-OSPO1-0-060413 DAL-SW-OSPOl -DB-060413 DAL-SW-OSPOI-LS-060413 DAL-SW-OSPO1-HS-060413 Amount Spiked (nglL) 1500 10500 500 500 500 1000 10000 1500 10500 500 500 500 1000 10000 1500 10500 500 500 500 1000 10000 1500 10500 500 500 500 1000 10000 1500 10500 500 500 500 1000 10000 600 1500 500 500 500 100 1000 "c-PFOA Amount Recovered (ngW Recovery (%) 1200 80 8240 78 389 78 389 78 429 86 509 51 4680 47 1240 83 8430 80 421 84 450 90 433 87 499 50 5300 53 1230 82 8100 77 41 9 84 420 84 451 90 589 59 6300 63 NR NR NR NR NR NR NR NR NR . NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR 56 1 94 1250 83 479 96 477 95 440 88 78.3 78 650 65 NR = Not reported due to quality control failures, see Table X for re-extraction data. Note: Since this summary table shows rounded results, recoveryvalues may vary slightly from the values in the raw data. Exygen Research Page 46 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Table IX. Surrogate Spike Recovery of 13CPFOA in Water Samples Continued Exygen ID C0171071 Spk C C0171071 Spk D C0171071 C0171071 Rep C0171072 C0171073 COI71074 C0171075 Spk E C0171075 Spk F C0171075 C0171075 Rep COI71076 COI 71077 C0171078 C0171079 Spk G C0171079 Spk H C0171079 C0171079 Rep C0171080 C0171081 C0171082 C0171083 C0171084 Spk C C0171084 Spk D C0171084 C0171084 Rep C0171085 C0171086 C0171087 C0171088 Spk E C0171088 Spk F C0171088 C0171088 Rep C0171089 C0171090 C0171091 C0171092 Spk G C0171092 Spk H C0171092 C0171092 Rep C0171093 COI71094 C0171095 Sample Description DAL-SW-OSP02-0-060413 DAL-SW-OSPO2-0-060413 DAL-SW-OSP02-0-060413 DAL-SW-OSP02-0-060413 DAL-SW-OSP02-DB-060413 DAL-SW-OSP02-LS-060413 DAL-SW-OSP02-HS-060413 DAL-SW-OSP03-0-060413 DAL-SW-OSP03-0-060413 DAL-SW-OSP03-0-060413 DAL-SW-OSP03-0-060413 DAL-SW-OSP03-DB-060413 DAL-SW-OSP03-LS-060413 DAL-SW-OSP03-HS-060413 DAL-GW-BPWELL-0-060413 DAL-GW-BPWELL-0-060413 DAL-GW-BPWELL-0-060413 DAL-GW-BPWELL-0-060413 DAL-GW-BPWELL-DB-060413 DAL-GW-BPWELL-LS-060413 DAL-GW-BPWELL-MS-0604413 DAL-GW-BPWELL-HS-060413 DPWS-WWI-DCTPOI-0-060413 DPWS-WWI-DCTPOI-0-060413 DPWS-WWI-DCTPOI-0-060413 DPWS-WWI-DCTPOI-0-060413 DPWS-WWI-DCTPOI -DB-060413 DPWS-WWI-DCTPOI-LS-060413 DPWS-WWI-DCTPOI-HS-060413 DPWS-WWE-DCTPOI-0-060413" DPWS-WWE-DCTPOI-0-060413" DPWS-WWE-DCTPOI-0-060413" DPWS-WWE-DCTPO1-0-060413" DPWS-WWE-DCTPOI-DB-060413 DPWS-WWE-DCTPO1-LS-060413 DPWS-WWE-DCTPOI-HS-060413 DAL-WW-EFF01-0-060413 DAL-WW-EFF01-0-060413 DAL-WW-EFF01-0-060413 DAL-WW-EFF01-0-060413 DAL-WW-EFFOI-DB-060413 DAL-WW-EFF01-LS-060413 DAL-WW-EFFOI-HS-060413 Amount Spiked (nglL) 600 1500 500 500 500 100 1000 600 1500 500 500 500 100 I000 10500 1000500 500 500 500 10000 100000 1000000 10500 100500 500 500 500 10000 100000 10435 100435 435 435 500 10000 100000 1500 10500 500 500 500 1000 10000 "C-PFOA Amount Recovered (ng/L) Recovery (%) NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR 8680 83 96200 96 391 78 393 79 426 85 4600 46 42500 43 8260 79 92400 92 343 79 350 80 391 78 4870 49 48800 49 1180 79 7880 75 360 72 335 67 425 85 744 74 6690 67 NR = Not reporteddue to qualitycontrol failures, see TableX for re-extractiondata. Note: Since this summary table shows rounded results, recoveryvalues may vary slightly from the values in the raw data. Exygen Research Page 47 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Table IX. Surrogate Spike Recovery of 13CPFOA in Water Samples Continued ExYgen ID Sample Description Amount Spiked (nglL) lac-PFOA Amount Recovered Recovery (nglL) (%) C0171096 Spk C DAL-LCH-MCLFO1-0-060414 10500 NR NR C0171096 Spk D DAL-LCH-MCLFO1-0-060414 100500 NR NR C0171096 DAL-LCH-MCLFO1-0-060414 500 NR NR C0171096 Rep DAL-LCH-MCLFOl-0-060414 500 NR NR C0171097 DAL-LCH-MCLFO1-DE-060414 500 NR NR C0171098 DAL-LCH-MCLFO1-LS-060414 10000 NR NR C0171099 DAL-LCH-MCLFOl -HS-060414 100000 NR NR C0171100 C0171101 C0171102 DAL-GW-TRIPl-0-060412 DAL-GW-TRIP1-LS-060412 DAL-GW-TRIP1-HS-060412 500 41 2 82 1000 748 75 10000 8150 82 C0171103 C0171104 C0171105 DAL-WW-TRIP1-0-060413 DAL-WW-TRIP1-LS-060413 DAL-WW-TRIP1-HS-060413 500 50 1 100 1000 808 81 10000 01 30 81 Average: 75 Standard Deviation: 26 A Sample was bottle was filled to 230 mL. causing the 13CPFOA spiking level to be modified. NR = Not reported due to quality control failures, see Table X for re-extractiondata. Note: Since this summary table shows rounded results, recovery values may vary slightly from the values in the raw data. Exygen Research Page 48 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Table X. Surrogate Spike Recovery of 13CPFOA in Re-extracted Water Samples Exygen ID C0169341 C0169345 C0169347 Spk E C0169347 C0169349 C0170984 C0170986 C0170989 Spk G C0170989 C0170990 C0170997 Spk E C0170997 C0171021 Spk E CO 171021 C0171035 COI71038 C0171060 C0171062 C0171063 C0171065 C0171071 C0171074 C0171076 Spk G CO 17 1076 C0171079 C0171083 C0171097 C0171099 Sample Description DAL-GW-I 38R-0-060412 DAL-GW-138R-HS-060412 DAL-GW-138s-DB-060412 DAL-GW-138s-DE-060412 DAL-GW-138s-MS-060412 DAL-GW-601R-0-060413 DAL-GW-601R-LS-060413 DAL-GW-601S-DB-060413 DAL-GW-601S-DB-060413 DAL-GW-601S-LS-060413 DAL-GW-601L-DB-060413 DAL-GW-601L-DB-060413 DAL-SW-BPPOI-DB-060413 DAL-SW-BPPOI-DB-060413 DAL-SW-BCTOI-0-060412 DAL-SW-BCTOI-HS-060412 DAL-SW-BC04-DB-060414 DAL-SW-BC04-HS-060414 DAL-SW-BC05-0-060414 DAL-SW-BC05-LS-060414 DAL-SW-OSP02-0-060413 DAL-SW-OSP02-HS-060413 DAL-SW-OSP03-DE-060413 DAL-SW-OSP03-DB-060413 DAL-GW-BPWELL-0-060413 DAL-GW-BPWELL-HS-060413 DAL-LCH-MCLFOI-DB-060414 DAL-LCH-MCLFOI-HS-060414 Amount Spiked (nglL) 13C-PFOA Amount Recovered (nglL) Recovery (Ym) 500 NR NR 1000000 NR NR 100500 NR NR 500 NR NR 100000 NR NR 500 639 128 100 114 114 600 777 130 500 772 154 100 106 106 10500 14600 139 500 540 108 1000500 1180000 118 500 243 49 500 475 95 10000 6850 69 500 432 86 10000 7270 73 500 607 121 1000 761 76 500 669 134 1000 641 64 1500 1590 106 500 630 126 500 341 68 1000000 230000 23 500 398 80 100000 53600 54 Average: 97 Standard Deviation: 34 NR = Not reported due to quality control failures. Note: Since this summary table shows rounded results, recoveryvalues may vary slightly from the values in the raw data. Exygen Research Page 49 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Table XI. Surrogate Spike Recovery of 13CPFOA in Sludge Samples Exygen ID CO171106 C0171106 Rep C0171106 Spk C CO171106 Spk D Sample Description DPWS-SL-DCTPO1-0-0000 DPWS-SL-DCTPOl-0-0000 DPWS-SL-DCTPO1-0-0000 DPWS-SL-DCTPOI-0-0000 Amount Spiked (ng/g) 4 4 4 400 I3C-PFOA Amount Recovered Wet Weight 1.82 1.67 1.48 291 Recovery (%I 46 42 37 73 Average: 49 Standard Deviation: 16 Note: Since this summary table shows rounded results, recoveryvalues may vary slightly from the values in the raw data. Exygen Research Page 50 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.:PO000760 Table XII. Surrogate Spike Recovery of 13CPFOA in Sediment Samples fiygen ID C0171107 C0171107 Rep C0171107 Spk C C0171107 Spk D C0171108 C0171108 Rep C0171108 Spk E CO171108 Spk F Sample Description DAL-SD-OSPOI-0-0000 DAL-SD-OSPOI-0-0000 DAL-SD-OSPOI-0-0000 DAL-SD-OSPOI-0-0000 DAL-SD-OSP02-0-0000 DAL-SD-OSP02-0-0000 DAL-SD-OSP02-0-0000 DAL-SD-OSP02-0-0000 NR = Not reported due to quality control failures. Amount Spiked (nglg) "c-PFOA Amount Recovered (nglg) Recovery (%) 4 NR NR 4 NR NR 4 NR NR 400 NR NR 4 NR NR 4 NR NR 4 NR NR 400 NR NR Exygen Research Page 51 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Table XIII. Total Percent Solids for Sludge and Sediment Samples Exygen ID C0171106 C0171107 C0171108 Sample Description DPWS-SL-DCTPOI-0-0000 DAL-SD-OSPOI -0-0000 DAL-SD-OSP02-0-0000 Total Solids (%) 22.83 58.39 53.33 Exygen Research Page 52 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 FIGURES Exygen Research Page 53 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Figure 1. Typical Calibration Curve for PFOA in Reagent Water 0428068 P760-1131 Water.rdb(PFOA): "Lineat' Regressionr 1 / $ weighting): y = 3.77e+003 x + 0.042 ( I = O.QQ82) 3.8e6 - 3.6e6 3.4e8 - 3.2e6 3.0e6 - 2.8e6 - 2.6e6 - 2.4e6 - 2.2e6 ~ Ln c c 3 2.0e6 ~ 0 u - m- 1.8e6 E - c 1.Be6 - 1.4e6 - 1.2e6 l.Oe8 - 8.0e5 8.0e5 4.0 e5 - 2 . 0 ~ 5 0.0 - 0 100 200 300 400 500 600 700 800 goo Concentration. nglL 1000 Exygen Research Page 54 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Figure 2. Extracted Standards of PFOA in Reagent Water, 25 ng/L and 50 ng/L, Respectively - XCO57006-7 PFOA Ftpndard) 47ML?620 amu -sample 2 of 38 front 042806B.wiR Area: 720787 covndp Wight: 7.67e+003cps R7: 77.5 mi# 1.00e4 - 1151 (o 8000.00- & u 6000.00 - 2000.00 - 0 . 0 0 - 1 - , .I . I 1 2 3 ~ I ~ I ~ II . ~I . tI . #~ ' I I . I ~. I .I 1 . ~ 4 5 6 7 8 Q 10 11 12 13 14 15 16 17 1 - Time, min XCO51008-2. PFOA(Standard)413.01389.0 arnu sample 3 of 38 from 042808B.wiff Area: 237159 counts Height: 1.48e+004 cps R T : 11.5 min 11.51 0.0 1 2 3 4 5 6 7 8 Q 10 11 12 13 14 15 16 17 Time. min Exygen Research Page 55 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Figure 3. PFOA in Reagent Control, 50 ng/L Fortified Reagent Spike A, and 500 ng/L Fortified Reagent Spike B, Respectively I - Reagent Control PFOA M k n o w n ) 47Z0.0610 amy -sample 9 of38 frorrr 042806B.wif7 @ e r r not found) 3000 2 v) Q 0 1 0' 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 Time, min Reagent Spk A . PFOA(QC)413.0#69.0 dmu .sample 10 of 38 from 042806B.wiff Area: 209542 counts Height 1.4Qe+004cps RT: 11.5 min 1.5e4- 0 1.0e4- - (cu 5000.0 -c c 0.55 0.0 1.63 3.88 4.73 5.97 - 8.08 15.00 16.Q2.17.39 0) 1.0e5- F0 ) c 5.0e4- c (u - c 0 .o 11.51 1 2 3 4 5 6 7 8 g 10 11 12 13 14 15 16 17 Time. min Exygen Research Page 56 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Figure 4. Chromatogram Representing a Water Sample Analyzed for PFOA (Exygen ID: C0169336, Data Set: 042806B) I - C0769336 PFOA (thkrown)473.On620 PMY -08lWPh 20 O f 3 8 from 0428068.wift Area: 824652 conif& Might: S85r+004cpo RT: 77.5 miif 1 5.584- - 5.0e4 4.584 - 4 . 0 ~ 4- uI 3.5e4- P Y $ 3.084: c al -c c 2.5e4 - I 2.004- 1.5~4- - I.Oe4 - 5000.0 0.59 0.0 1 I 2 3 4 5 6 7 8 Q 10 11 12 13 14 15 16 17 Time, min Exygen Research Page 57 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Figure 5. Chromatogram Representing a Sludge Sample Analyzed for PFOA (Exygen ID: C0171106, Data Set: 050906BR) 2.4e4 I I '3 2.2e4 2.0e4 1.Be4 1.6e4 pv) Q 1.4e4 u 1.2e4 -a, c c 1.0e4 8000.0 6000.0 4000.0 7 L 2000.0 0.0 1 2 3 4 5 6 7 8 Q I O 1; 12 13 14 I 5 I b I? Exygen Research Page 58 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Figure 6. Chromatogram Representing a Sediment Sample Analyzed for PFOA (Exygen ID: C0171108, Data Set: 051106A) - CO171708 PFOA (lhkrrown)4730/3620 ame -rarrrpte 21 of 24from 051106A.wif7 Area: 270507coentr Wight: 1.47e+OO4cp+ RT: 7 a 8 m i n 7 1.4e4 1.3e4 1.2e4 l.le4 1 .Oe4 QOOO.0 CaD u 8000.0 .$2_ 7000.0 -c c 6000.0 5000.0 4000.0 3000.0 2000.0 - 1000.0 10.32. I Time. min Exygen Research Page 59 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Figure 7. Typical Calibration Curve for 13CPFOA in Reagent Water 0428068 P760-1131 Water.rdb (13C PFOA): "Linear" Regression c1 /*'weighting): y = 3.86e+003 x + 0.0168 (I = O.QQQ4) 0 IO0 200 300 40 5d0 600 Exygen Research Page 60 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Figure 8. Extracted Standards of 13CPFOA in Reagent Water, 25 ng/L and 50 ng/L, Respectively I - XCO57006-7 73C PFOA (Standard]471M371LOamu -tample 2 of 38 frorrr 042806B.wifl Area: 702370 counts thiqht: &59+003 cpr RT: $7.5 min 2 8000 ,,, 6000 Q u 3w 4000 c a, -c c 2000 0 1 2 3 4 5 6 7 8 9 10 11 I - Time. min XCO51006-2 13C PFOA(Standard)415.01370.0 amu. sample 3 of 38 from 042806B.wiff Area: 212002 counts Height: 1.77e+004 cps RT: 11.5 min L_ 12 13 14 15 16 17 1.5e4- w nu . - 1.Oe4 5 c a, c c - 5000.0- 0.0'. I . I . I . I . I - I . 8 . I . I . 1 . I% - I . I . I . I . I . Exygen Research Page 61 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Figure 9. 13C PFOA in Reagent Control, 50 ng/L Fortified Reagent Spike A, and 500 ng/L Fortified Reagent Spike B, Respectively I - Reagent Control 73C PFOA &hrk$town]4fSW37aOamu -sample 9 of 38 from 042806B.wiff $eak not found) 60 (D D. Y 2.35 11.27 0- 1 2 3 4 5 6 7 8 Q 10 11 12 13 14 15 16 17 I Time, min Reagent S p k A - 13C PFOA(QC)415.01370.0 amu. sample 10 of 38 from 042806B.wiff Area: 207074 counts Height: 1.73et004 cps RT: 11.5 rnin 1.5e4 11.53 c m 5000.0 - c c 0.0 - 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 I Time. min Reagent Spk B. 13C PFOA (QC)415.0rj70.0 arnu. sample 11of 38 from 042806B.wiff Area: 1932996 counts Height 1.56e+005 cps RT: 11.5 min g-c m c c Time, min Exygen Research Page 62 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Figure 10. Chromatogram Representing a Water Sample Analyzed for 13CPFOA (Exygen ID: C0169336, Data Set: 042806B) I - e0169336 13cPFOA &knnrown) 411m7aom u -$8Wpfi! 20 of 38from 0428068.wiff Are8: 1595574courb l-&ig&t: 1.34?+005 cps R E 71.5 mitt 1 1.3e5 1.2e5 1.1~5 1.0e5 Q.Oe4 8.0e4 In Q 0 g 7.0e4 c -c 6.Oe4 5.0e4 4.0e4 3.0e4 2.0e4 1 .Oe4 0.0 1 L 1 2 3 4 5 6 7 8 Q 10 11 I 2 13 14 15 16 17 Time. min Exygen Research Page 63 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Figure 11. Chromatogram Representing a Sludge Sample Analyzed for 13CPFOA (Exygen ID: C0171106, Data Set: 050906B) I - C0171106 13cPFOA mknow@ 4ism7aoamv -+8mple f 6 of 2 1from 050906B.wiff Area: 131528covnb Height: f.73e+004cpt RT: 1aZmirr l.lOed 1 `3 1.OO eL oooo.oa 8000.00 7000.00 cn g Q u 6000.00 c 0 E- 5000.00 4000.00 3000.00 2000.00 1000.00 0.00 1 2 3 4 5 6 7 8 Q 10 11 12 13 14 15 16 17 Time, min Exygen Research Page 64 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Figure 12. Chromatogram Representing a Sediment Sample Analyzed for 13CPFOA (Exygen ID: C0171108, Data Set: 051106A) I - C0777208 73C-PFOA (Ulk~OWrr)4710/37a08MU-$aMple 27 of 24froM 057706A.wiff Area: 306027 covrrts might: 7.89~+004cprRT:l a 8 mirr 1.8e4 1.7e4 1.Be4 1.5e4 1.4e4 1.3e4 1.2e4 cn l . l e 4 P E 1.0e4 .- gooo.0 a, c G 8000.0 7000.0 6000.0 5000.0 4000.o 3000.0 2000.0 1000.0 0 .o `7 Time. rnin Exygen Research Page 65 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 APPENDIX A Study Protocol PO000760 (Exygen Study No. P0000760) with Analytical Methods and Protocol Amendments Exygen Research Page 66 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 STUDY PROTOCOL Study Title: Analysis of Perfluorooctanoic Acid (PFOA) in Water, Soil, Sediment, Fish, Clams, Vegetation, Small Mammal Liver and Small Mammal Serum Using LC/MS/MS for the 3M Decatur Monitoring Program Exygen Protocol Number: PO000760 Performing Laboratory: Exygen Research 3058 Research Drive State College, PA 16801 Phone: (814) 272-1039 Sponsor ReDresentative: Michael A. Santoro Director of Regulatory Affairs 3M Building 0236-01-B- 10 St. Paul, MN 55144 Phone: (65 1) 733-6374 -- Exygen Research Page 67 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.:PO000760 Exygen Protocol Number: PO000760 DISTRIBUTION: 1) JaisimhaKesari,Study Director, Weston Solutions 2) JohnM. Flaherty, Principal Investigator, Exygen Research 3) Michael A. Santoro, SponsorRepresentative, 3M Company 4) Exygen Research Quality Assurance Unit Page 2 of 65 -~ - - Exygen Research Page 68 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 F PROTOCOL APPROVAL Study Title: Analysis of Perfluorooctanoic Acid (PFOA) in Water, Soil, Sediment, Fish, Clams, Vegetation, Small Mammal Livers and Small Mammal Serum Using LC/MS/MS for the 3M Decatur Monitoring Program Exygen Protocol Number: PO000760 APPROVALS c QAhhL J a i s i 6 Kesari, Study Director Weston Solutions w &4?ka Michael A. S d r o ,-Sp.onsorRepresentative ' 3M Company $hn M. Flaherty, Principal Investigator / Exygen Research r sident, Facility Management MA&Date w g e n Research L/ " ~. Page 3 of 65 Exygen Research Page 69 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 TABLE OF CONTENTS TITLE PAGE ........................................................................................................ DISTRIBUTION....... ....................................................... 2 PROTOCOL APPROVAL ............................................................................................ 3 TABLE OF CONTENTS.................................................................................................................................. 4 INTRODUCTION............................................................................................................................................ .5 TEST MATERIAL ......... ............................................................. 5 SURROGATE FIELD SPME COMPOUND................................................................................................... 5 OBJECTWE ............................................................. TESTING FACILITY..... ........................................................ 6 STUDY DIRECTOR........................................................................................................ ...................... 6 SPONSOR REPRESENTATIVE ............................................................................................... 7 PRINCIPAL INVESTIGATOR ............... ............................................. 7 PROPOSED EXPERIMENTAL START ..................................................... 7 IDENTIFICATION AND SAMPLEPROCUREMENT, RECEIPTAND RETENTION ......................................................................... 8 SAMPLEIDENTIFICATION.......................................................................................................................... 8 ANALYTICALPROCEDURE SUMMARY .......... VERIFICATION OF ANALYTICAL PROCEDURE................ .................................................... 9 METHOD FOR CONTROLOF BIAS ............................................................................................................. 1 1 STATISTICALMETHODS ....................................... ......................................................................................................... I 1 REPORT .................................................................. SAFETYAND HEALTH ....................................... ............................................. 12 AMENDMENTS TO PROTOCOL ........................ DATA RECORD KEEPING .................................. QUALITY ASSURANCE ................................................................................................ 14 RETENTION OF DATA AND ARCHIVING ................................................................................................. 14 APPENDIX I. ANALYTICALMETHODS..................................................................................................... I5 Page 4 of 65 Exygen Research Page 70 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 - INTRODUCTION Exygen Protocol Number: PO000760 The purpose of this study is to perform analysis for perfluorooctanoic acid (PFOA) in water, soil, sediment, fish, clams, vegetation, small mammal livers and small mammal serum using L C M S N S for the 3M Decatur Monitoring Program. The study will be audited for compliance with EPA TSCA Good Laboratory Practice Standards 40 CFR 792 by the Quality Assurance Unit of Exygen Research. TEST MATERIAL The test material is perfluorooctanoic acid (PFOA) and was purchased from Sigma-Aldrich. PFOA Chemical Name: Pduorooctanoic acid Molecular Weight: 414 Lot Number: 23116HB Purity: 97.64% Transitions Monitored 413 -+ 369 (for quantification) and 413 -+ 219 (forconfimation) Structure: SURROGATEFIELD SPIKE COMPOUND The surrogate field spiking compound is I3C labeled periluorooctanoic acid (I3CPFOA) and was purchased by 3M from Perkin-Elmer Life and Analytical Sciences. "C PFOA Chemical Name: 1,2-13C perfluorooctanoic acid Molecular Weight: 416 Lot Number: 3507-195 Purity: 97% Transition Monitored: 415 + 370 Page 5 of 65 Exygen Research Page 71 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 Lc OBJECTIVE The purpose of this study is to perform analysis for perfluorooctanoic acid (PFOA) in water, soil, sediment, fish, clams, vegetation, small mammal livers and small mammal serum for the 3M Decatur Monitoring Program using the current versions of the following Exygen analytical methods: VOOOl780: VOOO1781: V0001782: V0001783: VOOOl784: VOOOl785: VOOOl786: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Water by LCIMSIMS" "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Soil by LCIMSIMS" "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Sediment by LCIMSIMS" "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS" "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Vegetation by LCIMSIMS" "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Small Mammal Liver by LC/MS/MS" "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Small Mammal Serum by LC/MS/MS" TESTING FACILITY Exygen Research 3058 Research Drive State College, PA 16801 Phone: (814) 272-1039 STUDY DIRECTOR Jaisimha Kesari P.E., DEE Weston Solutions, Inc. 1400 Weston Way West Chester. PA 19380 Phone: (610) 701-3761 Fax: (610) 701-7401 j.kesari@westonsolutions.com - Page 6 of 65 Exygen Research Page 72 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 SPONSOR REPRESENTATIVE Michael A. Santoro 3M Company Director of Regulatory Affairs 3M Building 0236-01-B-10 St. Paul, MN 55144 Phone: (651) 733-6374 PRINCIPAL INVESTIGATOR John M. Flaherty Exygen Research 3058 Research Drive State College, PA 16801 Phone: (814) 272-1039 - john.flaherty@exygen.com PROPOSED EXPERIMENTAL START AND TERMINATION DATES It is proposed that the analytical portion of this study be conducted from October 01, 2004 to December 31, 2005. The actual experimental start and termination dates will be included in the final report. IDENTIFICATIONAND JUSTIFICATIONOF THE TEST SYSTEM The following are the test systems for this study: Water (groundwater and surface water) Soil Sediment Fish Clams Vegetation Small Mammal Liver Small Mammal Serum The samples will be collected by Weston Solutions. The control samples will be purchased and prepared by the testing facility. Purchase and processing Exygen Research Page 73 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 details for the control samples will be included in the final report associated with this study. The test systems were chosen to access the environmental impact of PFOA in the Decatur, Alabama area. SAMPLE PROCUREMENT, lUK!EIPT AND RETENTION Water, soil, sediment, fish, clam, vegetation, small mammal liver and small mammal serum samples will be received at Exygen directly fiom Weston Solutions. The details of sample procurement for this study are outlined in the 3M work plan entitled "Phase 2 Work Plan for Sampling Environmental Media." The number and types of samples collected will vary depending availability in the field. The total number of samples received and analyzed for each matrix will be documented in the final report associated with this study. Water, soil, and sediment samples will be used as received without further processing at Exygen. These samples will be stored refrigerated at 2"C-SOC. Fish, clam, vegetation and small mammal liver samples will be processed according to the appropriate analytical method (see Appendix I). These samples will be stored fiozen at 5 -10C. Small mammal whole blood samples will be centrihged in the field at the time of collection and the serum - fiaction will be used for the study. Small mammal serum will be stored frozen at 5 10C. The receipt and processing of the samples will be documented in the final report and raw data associated with the study. SAMPLE IDENTIFICATION Prior to analysis, each sample will be assigned a laboratory sample reference number. The reference number will be unique and will distinguish each laboratory sample that is processed throughout the analytical procedure. Chromatographic data will be identified by the laboratory sample reference number. Sample storage conditions and locations will be documented throughout the study. Page 8of 6s Exygen Research Page 74 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 ANALYTICAL PROCEDURE SUMMARY References: VO00l 780: V000178 1: V0001782: V0001783: V0001784: V0001785: VOOOl786: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS" "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Soil by LCMSMS" "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Sediment by LC/MSIMS" "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Fish and Clams by LCIMSIMS" "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Vegetation by LCIMSIMS" "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Small Mammal Liver by LCMSIMS" "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Small Mammal Serum by LC/MS/MS" The above methods use analytical conditions capable of separating the isomers of PFOA. The final report will include the isomers summed into total PFOA found. VERIFICATION OF ANALYTICAL PROCEDURE A laboratory control sample will be used for the preparation of fortified control samples. The test substance will be made into solutions as per the method, and added to the matrices via a micropipette. For water sampling, Exygen will supply one bottle per sample collected. The bottles will be 500 mL preclean4 Sci/Spec Premier wide mouth HDPE bottles. These bottles have been routinely used for PFOA sample collection at the testing facility and have been shown to be free of PFOA. All containers used for water sample collection will be shipped to the sample location containing 100 ng of I3CPFOA (equivalent to 500 ng/L in the final sample). Samples will be added to each container to a volumetric fill line at 200 mL. A field duplicate, a low field spike and a high field spike of each sample will be collected. The low and high field spikes will contain PFOA as well as perfluorobutanesulfonate (PFBS), perfluorohexanesulfonate (PFHS), and perfluorooctanesulfonate (PFOS). These compounds are included in the solutions used to spike the samples. The results for PFBS, PFHS and PFOS will not be reported in this study. Exygen will supply one field blank (control water) and two field blank spikes (control water fortified with PFOA at a low Exygen Research Page 75 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 and high level) for every twenty samples collected. At the testing facility, each water sample (excluding field duplicates and field spikes) will be extracted in duplicate and will also be fortified at a low and high concentration with PFOA and processed through the described procedure to determine method accuracy and to check for bias. For soil, sediment, clams, and vegetation, Exygen will supply one 500 mL precleaned Sci/SpecPremier wide mouth HDPE bottle per sample collected or a zip-seal bag. All containersbags used for sample collection will be shipped to the sample location. Samples will be added to each container or bag in the field. At the testing facility, each sample will be extracted in duplicate and will also be fortified at a known concentration with PFOA at both a low and high level and processed through the described procedure to determine method accuracy and to check for bias. "C-PFOA will also be added to each sample in the laboratory prior to extraction at the level specified below. For small mammal liver, Exygen will supply a 50 mL polypropylene centrifuge tube. For small mammal serum, Exygen will supply a collection kit for each sample containing serum separator tubes (red top), vacutainers, needle holders and needles, transfer pipettes, and polypropylene tubes. At the testing facility, each liver and serum sample will be extracted in duplicate and will also be fortified at a known concentration with PFOA at both a low and r" high level and processed through the described procedure to determine method accuracy and to check for bias. '.'C-PFOA will also be added to each sample in the laboratory prior to extraction at the level specified below. I Low and high spiking levels for each matrix are defined below: Matrix I LowPFOA I HighPFOA I "C-PFOA Recoveries are anticipated to be between 70% and 130% of the fortified levels; however, the exact precision and accuracy will be determined by the analysis of the quality control samples described above. A statement of accuracy will be included in the final report. Prior to sample analysis, a laboratory control spike study will be conducted in water to justify the use of I3C PFOA results to establish precision and accuracy of PFOA samples. The following samples are to be prepared: Page 10 oJ65 - . Exygen Research Page 76 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 Two control water blanks Two control water samples fortified at 250 ng/L with "C PFOA Two control water samples fortified at 250 ng/L with "C PFOA and at 250 ng/L with PFOA. Two control water samples fortified at 5000 ng/L with "C PFOA and at 5000 ng/L with PFOA. The above samples are to be extracted and analyzed according to method V0001780 entitled "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS." METHOD FOR CONTROL OF BIAS Control of bias will be addressed by taking representative sub-samples from a homogeneous mixture of each matrix from untreated control samples, and by analyzing at least two levels of fortifications. STATISTICAL METHODS Statistics will be limited to those specified in the subject methods and to the calculation of average recoveries, as applicable. GLP STATEMENT All aspects of this study shall be performed and reported in compliance with EPA TSCA Good Laboratory Practice Standards 40 CFR 792. The final report or data package (supplied to the Sponsor) shall contain a statement that the study was conducted in compliance with current and applicable GLP standards and will outline any deviations in the study from those standards. This statement will be signed by the Study Director and Sponsor Representative. REPORT A final report will be prepared by the principal investigator or their designee at the conclusion of the study. The report will include, but will not be limited to, the following: Page I I of65 - Exygen Research Page 77 of 140 ~~~~~~ ~~ Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygcn Protocol Number: PO000760 The name and address of the Study Director, Sponsor Representative, and of the testing facility. A statement of GLP compliance (any related documentation, such as chain-of-custodyrecords, must be in the study records). The signed and dated statement by the Exygen Research Quality Assurance Unit regarding dates of study inspections and dates findings were reported to the Study Director and Management. A description of the exact analytical conditions employed in the study. If the subject method was followed exactly, it is necessary to include only a copy of the analytical method. Any modifications to this method will be incorporated into the report. If the method is photo-reduced, the project number and page number must be included on each page. Description of the instrumentationused and operatingconditions All results from all sets analyzed. Control and fortified samples will be identified and the data table will include sample number and fortification level. Representative chromatograms for each analyte in each matrix, including chromatograms of a standard and a control sample, and a chromatogram at a fortification level. The location of the analyte peaks will be clearly identified in all chromatograms. All circumstances that may have affected the quality or integrity of the data will be documented in the report. Locations where raw data and the final report are to be archived. Additions or corrections to the final report shall be in the form of an amendment signed by the Study Director. The amendment shall clearly identify that part of the report that is being altered and the reasons for the alterations. The amendment will be signed and dated by the Study Director and the Sponsor Representative. SAFETY AND HEALTH 0 Laboratory personnel will practice good sanitationand health habits. Every reasonable precaution shall be taken to prevent inadvertent exposure of personnel and the environment to the test or reference substance(s). Page 12 of 65 Exygen Research Page 78 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 r AMENDMENTS TO PROTOCOL All significant changes to the analytical protocol outlined here will be expressed in writing, signed and dated by the Study Director and Sponsor Representative. Amendments usually will be issued prior to initiation of study plan change. However, when a change is required without sufficient time for the issue of a written amendment, that change may be effected verbally with supporting documentation signed and dated by the Study Director and followed with a written amendment as soon as possible. In this case, the effective date of the written amendment will be the date of the documented change. Copies of the signed amendments will be appended to all distributed study plan copies. The original amendment will be maintained with the original study plan. Any deviations from the study plan or from the analytical method as provided will be documented and reported promptly to the Sponsor Representative. DATA RECORD KEEPING Records to be maintained include the following (as appropriate): 0 Sample tracking sheet(s) 0 Sample receipt records, storage history, and chains of custody 0 History and preparation of standards (stock, fortification,calibration) 0 Description of any modifications to the method 0 Instrument run sheets, bench-sheets or logs . Analytical data tables All chromatographic and instrumental conditions Sample extraction and analysis dates . A complete listing of study personnel, signatures and initials Chronological presentation of all study correspondence Any other documentationnecessary for the reconstruction of the study Chromatograms-All chromatogramswill contain the following: Sample identification, injection date, arrow or other indication of the area of interest, and injection number corresponding to the run. Additionally, fortifications will include the amount of analyte added and the sample number of the sample that was fortified. Analytical standard chromatograms will additionally include the concentration (e.g., @nL). Page I 3 of65 Exygen Research Page 79 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 P- As part of the documentation the following sheets will be included in each analytical set: a run sheet listing the samples to be run in the set, and an instrument conditions sheet describing the instrument type and operating conditions. All applicable requirements for reporting of study results as per 40 CFR 792.1 85. QUALITY ASSURANCE The QA Unit of Exygen Research will inspect the study at intervals adequate to assure compliance with GLP's. and will report the findings of audits to the StudyDirector, Exygen Management,and the SponsorRepresentative. RETENTION OF DATA AND ARCHIVING All hard copy raw data, including, but not limited to, the original chromatograms, worksheets, correspondence, and results shall be included with the data package submitted to the Study Director. These will be archived with the original study plan, amendments, final report, and all pertinent information from the Sponsor. The testing facility shall keep all electronic raw data and any instrument, equipment, and storage logs for the period of time specified in 40 CFR 792.195. An exact copy of the materials submitted to the study director will also be kept at Exygen Research. Exygen will obtain permission fiom the study director before discarding or returning samples. Exygen Research Page 14 of 6s Page 80 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PoooO760 APPENDIX I ANALYTICAL METHODS V0001780: VOOO1781: V0001782: V0001783: V0001784: VOOOl785: V0001786: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS' "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS" "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Sediment by LC/MS/MS" "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS" "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Vegetation by LC/MS/MS" "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Small Mammal Liver by LCMSMS" "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Small Mammal Serum by LCIMSIMS' Exygen Research Page IS of6S Page 81 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 ANALYTICAL METHOD MabodNumbcr: V0001780 Metbod of Andytb for the htermlnatlon of Ptrfluorooct.noic Acid (PFOA) in Water by LCIMSIMS AnalyticalT n t i n ~Flciiity: Exygen Ruurch 3058 R-h Drive StrteCollcge,PA 16801 Appmvcd By Vice Ruidcnl, Opartiom, Exyga Ruearch Exygen Research Page I 6 of 6s .- .. Page 82 of 140 Interim Report #24 - Analysis of Water, Sludge,and Sediment Exygen StudyNo.: PO000760 Exygen Protocol Number: PO000760 -&Yam Metbod Nwnbcr VOW1780 I 1 ANALYTICALMETHOD Method of Analysis for the Dczrmninationof PerIluorooctanoicAcid (PFOA) in Water by LCIMSIMS 1.0 scopc Tbii method is to bc employed for tbc bolation and quatitation of pcrfluomoctanoic acid by High Pafomuncc Liquid Chromatography coupled lo a tandem Mars SpecbumctricDetector (LUMSiMS) u1 wata. 2.0 safely 2. I A w a y observe d e lsbontorypracticer. 2.2 C o d t the rppmpriatc MSDS before handling m y chemical for pmpm safety P~tiOM. 3.0 SampleRcquircmmt 3. I At l a m 40mL of t a l ample for extraction. 3.2 No iamplc pmccuins,is Mdedfor water samplcr. 3.3 Sampled stored rcfngcnted ahould be allowed to equilibrate to r a m (a3np-. 3.4 All r m p l a nul bc thoroughlymixedbcforc being sampled for extraction. 3.5 Any u m p l a containin#patticla should be centrifuged at -3000 rpm for -5 minuta and the mpanr~nutrcd for the extraction. 3.6 Sample colleclion pmcedura will be apccified in lbe sampling plan for this pmject. 4.0 RcagenuandSundvdr 4.1 Wata-HPLC 4.2 Mahvlol-HPLC&~adc - 4.3 Ammonium -e A.C.S. Reagent Grade - 4.4 P d u o m a c ~ iAccid Sip-Aldrich 5.0 InreumcntdEquipmcnt 5.1 A hi@ pcrfomruref liquid h m t o g n p h capable of pumping up lo 2 lolventa oqu~ppcdrnth 8 vvieblc volume injector c.p.ble of injccnng 5-200 pL ~ M e c t o tdo 8 rnndcmM U SpCCholUCter (Lc/MS/MS). 5.2 A device to collectn w dit. for pwk integration and quatitstion. 5.3 AMlyliulb.lmcc capable of rcding to o.mo1 g. 5.4 50 mL dirpoaablcpolypmpylmc cmhifugetuber. 5.5 I5 mL diipoublcpolypmpylme centrifugetuba. 5.6 Disposableminopipar (SO-loOUr,lOO-ZOOuL). 5.7 125-mLLDPE numw-mouth bonlol. 5.8 2 mL clur HPLC vial kit. 5.9 Dirpoublcpipatcs. 5.10 AutopipaW(lOO-1OOOpLMd 10-100pL). wilhdirporablctips 5.11 W~tenScpPJ;Vr6cc(lg)tC18SPEcurridgea. Page I7 of 6S Exygen Research Page 83 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 f- Exygen Protocol Number: PO000760 Exyga Ruurch MelhodNvmberVMM1780 I 1 AhALYTlCAL ,METHOD Mschodof Asllysufor the Dacrminstionof P d w m o c w o i c Acid (PFOA) in Water by Lc/Ms/Ms 5.12 SPE m l u m maaifold 5.13 C c n t r i f u ~ e u p a b l c o f a p5O~ mLpolypmpylenetubat 3000rpm. 6.0 ChromltogrrphicS y ~ l a n 6.1 M y t i c a l Column: Flwplw RP (KeystoneScientific).2.1 nun x 50 mm.Sp (PN 82505-052130) 6.2 Tcm~uUtu'e:30.C 6.3 MobilePhuc (A): 2 mMAmmonium Accl.1~in Water 6.4 Mobile Phuc (B) : Mnhrnol 6.5 Gradient Fmlpsm: IbrJS6mu 0.0 65 1.0 65 8.0 25 20.0 25 22.5 65 Flow Rate ULPLELmial 35 0.3 35 0.3 75 0.3 75 0.3 35 0.3 - 6.6 InjectionVolume: IS pL (anbe i m p r e d to PI much PI 50 pL) 6.7 QuultiIAon: Pulr A m - exlcnul standard calibrationcurve. 6.8 Rua Time: 23 minutu. The abovec o d i t i a w intendedu D guide md may be changed in order to o p ( i m i the mu:ryrtan. 7.0 M S M System 7.1 Mode:ElectmaprayNegalivo MRhimode, monitoring 4 I3 + 369 dz The above conditio^ UT intended u a guide and may be changed in order IO optimizethe MSMS system. 8.0 Repantion of Solutiom 8.1 MobilePbc 8.1.1 2 mM ammonium &e in water ia prepared by adding 0.154 L( of v ~ ~ n i uawmtuc to IO00 mL of water. Altcmptc volumo m y be pnpsred. Exygen Research Page 1%of 65 Page 84 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 Exyprn Rarvch Method Nvmbn VwO1780 1 1 ANALYTICAL rnmioD Mctbodof ADslylis for the Detaninltion OfPerfluomtanoic Acid (PFOA) in Water by LC/MS/MS 9.0 SlsodprdRcpprptjon 9.1 Standad StocwFortification Solution 9.1.1 Rcpvc a stc& solutionof -100 p&L of PFOA by weighing IO mg of dfidNndud( ~ ~ ~ c fflorepdurity) urd dilute to 100 mL with mdh.aol in a 12s-mLLDPE bottle. 9.1.2 A IO p g h L fonificationwlution of PFOA is prcpsred by bnnginp 111 mL of thc IM) pB/mLwlution to L f i ~ vlolume of 100 with methanol in a 125 mL LDPE bottle. 9.1.3 A 1.0 p g h L fonificationwlution ofPFOA is prepared by bringing I o mLofUle IO pB/mL wlution to a final volume of 100 with msihanolt~i a 125 mL LDPE bottle. 9.1.4 A 0.1 p g h L fortificaion solutionof PFOA is prepared by bnnginp Ill mL of thc 1.O pg/mLsolutionto a final volume of I00with ~ t h r n oiln a 125 mL LDPE bottle. 9.1.5 A 0.01 p#mL fo8tificationsolution of PFOA is preparedby bringing IO mL of the 0.1 pdmL solution to a h l volume of 100 with mcchrol in a 12s mL LDPE bottle. 9.1.6 Ihsrtock d fortificationW ~ U ~ ~ aOrcMto be i t o d in a refrigerator ai appmximltsly 4'C urd arc [table for 8 maximum period of 6 months from the data ofpreprrPtion. 9.2 Sundud Calibration Solutions 9.2.1 LCMSMS calibration stpndudr arc pnppred in HF'LC water. The calibration ruodudr arc procured ulrou@~the extraction procedure. idantid to rmplu. 9.2.2 The following ir a typical a m p l e : additional conccnaationr may be prepared M needed. Fuvl Concmprrim Fatifiutim Volume of Conccnmtion of C.librni~on ofFuiif& Volume FortifiedContml Calibnum SmdnrdID Solution~wb) (Id,) S m l c (mL) St.ndrrd [upt)* (example) 0 0 40 0 XCmmddyy.0 IO 100 40 15 XCmmddyy.1 IO 200 40 50 XCmmddyy.2 IO 400 40 IW XCmmddyy-I 100 100 40 250 XCmmddyy.4 100 200 40 500 XCmmddyy-J 100 400 40 loo0 XCmmddyy-6 -'Thoatnc!ed concerhtion oftha dibration standard is equal to 8x its initial concCnmUon, due to thc sonscnmtion of the standard during the extraction(SPE) XC atnctsd calibration sundud. Page 19 of 65 Exygen Research Page 85 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 ExygenProtocol Number: PO000760 F- 9.2.3 A m u rtrrrdsrdsolution (rcagmt blank) with each ccl of ilmdubumsted. 9.2.4 StoreaIl cxlxackdsrlibntion rtudnrdr in 15-mL polypropylenetubes .I2'C to 6'C, up to two wccb. 9.2.5 AltanUc volumu md collccnhltio~of slandudr m y be prepared as needed. 10.0 BatchSctKJp 10.1 !5&batch of u m p l a extndod (typically 20 or krs) must include at least OM - a t contml (mctbod blank using HPLC water) and two reagent wntmla fortified at h o r n concmlntionr (lab control spike) to venf? p d u r a l recovery for tho batch. 10.2 RcpuirCmentafor field and I.bantory duplicatm and spikes will be specified in the quality auurulcc plan for thisproject. 11.o Sample Extnction 11.1 Mcaatuc 40mL of umpleor a p o r t i ~of rmplc diluted to 40 mL with water into 50 mL polypmpylene mlrihge Nbm (fortifyas needed. replace lid and mix well). 11.2 Condition tbc CII SPE Wvidga (I g, 6 mL) by pusing 10 mL methanol followedby 5 mL of HPLC water (- 2 droplra). Do not let column run dry - 11.3 Load ampleon conditionedCIISPE oareidgc. Discard eluate. 11.4 ElW with -5 mL 100% methanol. Collect 5 mL of eluate into graduated I5 mLpolypropylene cenhihpc Nbu(find volume 5 mL). I1.I AnaIya umpluusingolectmlpny LCIMSIMS. 12.0 Chronulopaphy 12.1 Injoet lhc umc mount of each slandud. runplc and fortifiedsample into the Lc/MS/Ms sylum. A calibration iund.rd must precede and follow a11 rnrlyLCd ,unplm. 12.2 Sunduda of PFOA-oc to .t least five or more concmtrauon levels must ba included in ID .nrlyticd act. 12.3 A0 enlire set of cxtnctcd calibration nurdudr must be included at the bagi@ning and .Ithc cnd of I umplc act. Extracted standards must k intmpcncd bawcea cvcy 5-10 aunplu. Aa M alternative. an entire sei 01' cxmelcd dibntion ctandard~may be injected at the beginning of a ret followed by extncLtd c.librocion rundrrdr intcrapersed every 5-10 samples (to account for I w n d sct of exhlcted slandds). In either case. cxvaclcd dibrptioo at.odudr must k the 6Rt .ndl ~injtection in a umple set. 12.4 UK iinur NDdvd c w u for qutituion. Linear standard curves are generuedfor lhc arulytc by linarrepmion uriog l/x weighting of peak area Exygen Research Page 86 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 E x y p Ruurcb Mnhod Nvmbn VMo1780 I I ANALYTICAL METHOD Mahod of Analysis for the -on of PCmuomoaMoic Acid (PFOA) in Water by Lc/Ms/MS v ~ l wcrlibmion Nndvdconcaotruionwing MarsLynx 3.3 (or quivalcnt) r o syltan~. ~ 12.5 Sunplc rapow should not exceed standard responses Any samples that ex& rt.ndud responsesahould be iintha diluted and reanalyzed. 13.0 Acccptanw criteria 13.1 ChrOmDtopm mwl &ow a peak of L drugblcr ion at 369 MU from a pueni of413 mu. The 413 MU parent coMponds to the PFOA anion. while ihe dauphtcr ion (369 MU) rcpreunu the lo66 ofcarbon dioxide. 13.2 Method blank: must no1conlain PFOA at levels greater than the LOQ. I f a blank conuiol PFOA aI Isvela pnlu than 50 n#L. then a new bImA sample mwbcobt.ined .adthe entireset must be m x m e t e d . 13.3 Rocovmicl of conml8piLcr ud rmeiX spike@mmut be between 70-130% of thir knownvalua. If a control apike falh outside the acceptable limits, h e enlire ad of n m p k ahould be rwrtncted. Any mahix spike outside 70- 13Vh lbwld be cvlluatcd by the uulyat to determine if re-extraction is WYlrnlCd. 13.4 Any d b n t i o n Nndvd found to be a atuisticd outlia by using the Huge Emf TuL m y be excluded from the calculation of the calibration cuwe. Howwcr, tbc total number of cxmctcd calibration standards that could be acludcd must MIex& Mated. 20% of the total number of cxtrscled standards 13.5 m* The oorrclrtion COcmCicnt (R) for 20.992 20.985). If calibration d d b n t ti r on fdl c o w utr u id pnaat e thsrs cd must be limiu. then appropriate steps must be taken lo adjust itumMent operation, and the lund.rdror the relevant IC(of umpla should be reanalyzed. 13.6 Retation limu between and rsmplcr must not drift more than f 4 K w i t h UI d y l i c d ND. If retentiontime drifl exceedsthis limil within an uulyliul run thcn the KImwt be runalyred. 14.0 Cdculrtiolu 14.1 U K UK followinp q u t i o n lo UlculUe the mount of PFOA found (in ng/L. brrcd on peak area) win# the standard curve (linear regreasion parameters) gmmatcdby the Mur Lynx softwarepmgrun: - PFOA found(npn) x DF slops -DF frtor by which the fuul volumew u diluted,if necessary. Page 21 of 65 Exygen Research Page 87 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 I I ANALYTICAL METHOD MdhodofAnJyrir for tbsDolcnninuion of Pu'fluorooctmoicAcid (PFOA) in Water by LC/Ms/MS 14.2 For wnplor forti6od with known mounu of PFOA prior lo cxlraclion. use the followiq equationto cdcul.tc thep e n t recovery. Rewvay (%) = Exygen Research Page 22 of 65 Page 88 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 Metbod ANALYTICAL METHOD MochodNumba: VooO1781 Analficd Testing Facility: ExygmR t s u n h 3058 R a u n h Drive Stale College,PA 16801 Approved By: Pull CoMOlly I Tahniul hda,LC-MS, Exygm R a u r c h In Soil by Exygen Research ToulPaga: 7 Page 23 of 65 Page 89 of 140 ~ ~ _ _ _ _ _ _ ~_~ ~~~ Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 Exygen HelbodNumbrrVOM)I7SI I i ANALYTICALMETHOD M c W of And@ for thc IMamirutionof PCmuomoctnnoicAcid (PFOA)in Soil by LC/MS/MS 1.o Scope This mabod b to be anployed for the iaolntionand qwtilation of perfluorooctanoic acid by High Perfomuux Liquid Chromatography coupled to a tandem Mass SpcstromctncDctcstor (Lc/MS/MS) in roil. 2.0 Sltcty 2.I Alwap oblorve u f c l h t o r y practicer. 2.2 Consult the approprialcMSDS before handling any chemical for proper safely precautions. 3.0 SampleRcquinmenc 3. I AI lcut 15 gof tu(m p l e for exmtion. 3.2 No ample pmccuing ir needed for soil sampler. 3.3 Sampla ltorcd rebigented should be allowed to equilibrate to mom tompcnhuc. 3.4 All ramplu mwf be ulomughlymixed before being sampled for extmclion. 3.5 Sample collection pmcedurra will be rpcfificd in the sampling plan for this projccr 4.0 Ragenu and Standuds - 4.1 W.Lor HPU:pndc 4.2 Mdhrnol-HPLCgrule - 4.3 AmmoniumAcd.tc A.C.S. Ruga11Grade - 4.4 P4uorooct.ooic Acid Signu-Aldrich 5.0 Insment and Equipment 5.1 A high p e r f o m liquid chromatograph capable of pumping up to 2 solvcnu equipped with a vuiable volumc injector capable of injecting 5-2W & omncslodto a 1.ndunM u r Spcctromota (LUMSMS). 5.2 A device to collect raw data for p u k inteption and quantitation. 5.3 Arulylic.lb.luuecapableofreldingto0.00001 g. 5.4 50 mL disposlblepolypropylenecentrifugetubes. 5.5 I5 mL dicporrblepolypropyleneccnnifuge tubes. 5.6 Dicporrblcmicropipetr(50-1ooul IW2oouL). 5.7 125-mLLDPEnurow-mouthbonlw. 5.8 2mLcleuHPLCvi.lkk 5.9 Dirpoublcpipetta 5.10 Autopipeuu (100-1OOO pL ud 10-100 pL). with dilpo8ablc tips 5.11 W.tmS~P~Vr6ee(Ig)tC18SPEoutridger. 5.12 SPE v.cuum auaifold. 5.13 Ultrvonicbr(b. Exygen Research Page 24 of 45 .. Page 90 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 %a=- MetbodNumberVW01781 1 i ANALYTICALMETHOD Methodof Arulpia forthc KkIaminalion of Pcrtlwmtanoic Acid (PFOA) in Soil by LC/Ms/MS 5.14 Wriat-lctionIh*cr. 5.I5 Centrifuge capable of spinning 50 mL polypropylcnc ~ b Cart SO00rpm. 6.0 Cluomato~hicSynQn 6.1 halycicll Column: Flwpbvs RF' (Kepbne Scientific), 2.1 mm x 50 mm. 5p (PM: 82505-052130) 6.2 Tcmpcnturc: 30-C 6.3 Mobile Phuc (A) : 2 mM Ammonium Acetate in Water 6.4 Mobile Pbvs (e) : MeUunol 6.5 GndientPmgnm: Flow Rate !&hllull& 65 35 0.3 1.o 65 35 0.3 8.0 25 75 0.3 20.0 25 75 0.3 22.5 65 35 0.3 6.6 6.7 6.8 - - InjectionVolume: I5 )tL( QurntiUtioa: P u k Am Run Time: 23 minutoc. C c U x bI e t inwurcd to d cundud d B( much M 50 pL) b n t i o n curve. Thc above Eonditio~Mintended PI Iguide and my be changed in order lo optimize thc HPLC rptem. 7.0 MS/MSSptcm 7.1 Mode: Electmapray Negative MRM mode, monitoring413 +369 m/r for PFOA. The.bovc c d t i o n r Mintcndcdu 1 guide and may be changed in order 10 optimize thc MSMS ryltm. 8.0 Rcp.ntion of soluciorv 8.1 MobilePhrc 8.1.1 2 mM unmonium acetate in water is p r c p d by adding 0.154 8 of d u m rctate b loo0 mLof water. AltpMtc volumsl m y be prepared. Page 25 of 6s Exygen Research Page 91 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 1 I ANALYTICALMETWOD Method of Annlyak for the L l e k m h i o n of PufluomocUnoic Acid (PFOA) in Soil by LC/MS/MS 9.0 S(udsrdReppntion 9.I Sundnrd StocWFortificationSolution 9.1.1 Rcrvro a stock solutionof -100 udmL of PFOA bv weinhinn IO my of k l y t i c d iundud(concctcd fGpurity) and dildc IO YO0 kL wgi; meUunol in a 1 2 5 4 . LDPE bottle. 9.1.2 A IO pghL fortification solution of PFOA is p r e p d by bringing I O mL of lhe 100 pP/mL solution to a fmrl volume of 100 with methanol m a 125 mL LDPE bottle. 9.1.3 A 1.0 )IglmLfortifiulion solution of PFOA is prepared by bnnging IO mL ofthe IO pg/mL solution to a final volume of 100 with methanal in a 125 mL LDPE bottle. 9.1.4 A 0.1 pghnL fortificationsolution of PFOA ia prepared by bringing IO mL of C 1.0 pP/mL solution to a fmal volume of 100 with m e t t ~in~ l a 125 mL LDPE bottle. 9.1.5 A 0.01 p g h L fortification solution of PFOA is prepared by bnnging IO mL of thc 0.1 &mL solution to a final volume of 100 with mNvnolin a 125 mL LDPE bottle. 9.1.6 Ths stack and fortificationsolution8PIC to be stored in a refrigeratorPI lppmximatcly 4'C and arc ruble for Imaximum pcriod of 6 months born the date o f p r e p d o n . 9.2 S U D MMibration Solutiotu 9.2.1 Lc/Ms/MS calibntion U a u d d ~PIC p'cpucd in HPLC water. The cdibntion standda arc proccued Ulrough the extraction procedure. identicalto umpla. 9.23 The following ir a typical example: additional concentrations may be prrpprcd u nscded. CcaatrUioo Fortificatioa Volums of Fmal Concentration of Calibration IO 100 40 25 XCmmddy;- I IO 200 40 50 XCmmddyy-2 IO 400 40 100 xcmmddyy-3 100 100 40 250 XCmddyy4 100 200 40 500 xcmmddyy-5 I00 400 40 loo0 XCmmddw-6 -The arm~tcdcowllh.tionof the cdibnt wncmtration,due (0 the EooMmfion of XC extnctcdcalibntion rt.nd.rd. ion thc tundud Madud is equal to during the 8x its initial extraction (SPE) Exygen Research Page 26 of 65 - - .. .. - Page 92 of 140 - ~~~~~~ Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 9.2.3 9.2.4 9.2.5 A zao cIandnrd solution (reagent blank) with each ret of sundprb extracted. Store all Cxtnctcdulibruion ctpndnrdr in 15-mL polypropylene rubes 2'C to 6'C. up to two we&. Altcmr(cvoluma urd CO~lceallfnOMof staadud6may be prepared as noodpd. 10.0 BuchSc~Up 10.1 Each bat& of rrmplu extracted (typically 20 or Ius) must include a1 least 01%rea@ mnhol (mahod blank Wing 5 mL of methanol)and two reagent conhull fonified u Lwm eoncpltntionr (lab control spike) to verify pmcedvnl m v a y for the batch. 10.2 Fkquiremcntr for field urd labontorydupliutu d spikes will be specified in the q d t y Y~YMEC plan for thisproject. 1I .o Sampleextraction 11.1 Wcigb 5 g of umplc into 10 mL polwopylcnc centrifuge tube6 (foflify as nccdod.r o p h lid admix well). 11.2 Add 5 mL of mcIhw1and hake on a Wrist action rhrker for -1 5 minuter. 11.3 T ~ f c r U w l u t u ( o u 1 u l t r w o i bs .thandronicitcfor-I5minutsr. 11.4 Bring cbc volume up to 40 mL with water in the SO mL polypropylene cenhifuge tubc. 11.5 ccsleifugefor-l0minutu.t -3ooorpm. 11.6 Condition the CII SPE oucridga ( I g, 6 mL) by passing 10 mL methanol followedby 5 mLOfW L C Water (- 2 drop/CCC). Do MI Icl column N n dry 11.7 Load (decant) the sample on the conditioned C,I SPE cartridge. Discard - elute. 11.8 Elute witb -5 mL 100% methawl. Collect 5 mL of eluale into graduated I5 mLpolypropylme centrifugeNbcs(Wvolume 5 mL) 11.9 Anrlyzc vmplol wing clectrolpny m S / M S . 12.0 chmmrtomby 12.1 Inject thc v m e YILDLUL( of e& ~undrrds.ample and fortified sample into the Lc/Ms/Ms iyaan. A ulibntion d u d must precede .nd follow all lrulyzul umpla. 12.2 Smbrb of PFOA sonclpwdino to u l u l l five or more cornamlion levels mudbe included in u1 uulytiod w. 12.3 An auirc ret of cxtncted calibration standuds must be included at the bc-g and 81 the end of a umplc (01. Extracted standards must be lnlarpensdbetween every 5-10 sampla. an altanntivc. an entire set of PI@ I of 7 Page 27of6S Exygen Research Page 93 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 I 1 .ANALYTICAL METHOD Method of Andyrir for the Daarminuiooof P d w r o o c w o i c Acid (PFOA)in Soil by Lc/Ms/MS extracted d i W n atsndsrb m y bo injected at the bcginning of a set followed by ex(nctcd c d i i d o n rtsndarda intarpencd every 5-10 samples (to mx~untfor a m n d set of extracted stadanla). In either case. extracted crlibntion slanddn mwt be thc fint and laat injection in a sample set. 12.4 Ute liwu rt.ndud c w u for quantitstion. Linear standard curves u e penmted forthe anrlytc by linear reerasion wing Ihr weighting of puk area v m ulikuion Nadudconcentration uing MppsLpx 3.3(or equivalent) toftarM ryrtam. 12.5 Sample rorpoluo ibwld not c x d standud rcsponeu. Any samples that exceed standardruporucishould be M u dilutedmd WWlyzed. 13.1 Chmnatogrmnmurt show a peak of a daughter ion u 369 MU From a parent of 413 mu. Tbc 413 M u pplaot correqonda to the PFOA Mion. while Ihe bughtuion(369m u ) rqnuenuthe IOU of carbon dioxide. 13.2 Mcthod b l a b mwt not contin PFOA at lcvelr greater than the LOQ. IF n blank contdna PFOA at lwelr p t m than 50 npn. then I new blank sample mwt be ob- rad thc cntircwt mwt bo nzxtncted. 13.3 RsoveJi~olf c o n h l apikm md mbix #pika must be between 70-130% of their h w n vdua. If a control lpike Wls outside the acceptable limits. the entire m of wmplu should be re-extracled. h y matrix spike outride 70. 13W. rhould be cvduated by the d y r t to detuminc if re-extraction is WuMLLd 13.4 Any dibnlion rt.adsrd found to be a stPtirtical outlier by wing the Huge Error TCn may be excluded fmm the dculation of the calibration curve However, the cwl number of cxtncted calibration rtmdarda that could be excluded mwt not exceed 20.h of the total number of extracted standards injected. 13.5 'Ihc cornlalion cocfficicnt (R) for calibration CIUVCII generated must be 20,992 (R' 20.985). If crlibntion rcrulu fall outtidc these limits. then 8ppropri.tC ncpi mwt be t.km to adjust inmumcnt operation. and the ttand.rdr or thc ~CICVUIImof umplu should be rednalyzed. 13.6 Ratention tima bstwosn ttsndud~rad umplu muit not drift more than f 4 K within UIw l y t i u l run. Ifracntion time driA cxceadathis limit within UI uulyticll IUII then thc wt muit bo reuulyLod. Exygen Research Page 28 of 65 Page 94 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 1 I B v l m M e M Number VMx)1781 ANALYTICAL.METHOD Method of ruulyxi~for thc Dstmninationof Pcrfluomoclanoic Acid (PFOA)in Soil b) LCIMSIMS 14.0 Calculations 14.1 Use the foUowing equation to cdculntcthe mounl of PFOA found (in nuL. b u d rm peak ma) &g tbo I(.nd.rd CIUVC (linur regreasion pusmeterr) - pcnontcdby thc Mur Lynx loflwmp m p m : PFOA found (#L.) @s!& ' ' tL x DF ,loPC DF = factor by which tbo fiollvolme w u dilulsd. if necessary 14.2 F a umplu fodficd with known u n o ~ l l Uof PFOA prior Io cxvaclion. LSC - the followingopuuiwIO k c o v e r y (%) wlcul.tc che percent recovery. 14.3 Use tbc following equuion to EonvcIt the mounl of PFOA found in ng/L 10 ndg Wb). PFOA found @pb)= umplc weight (5 g) 14.4 Use the fobwing eqwion (0 d c u h e the mounl of PFOA found in ppb bucdOD dry Weight. PFOA found (ppb) dry weight - PFOA found (ppb) x [ 100% / lolsl soli&(%)] Exygen Research Page 29 of 65 Page 95 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 ANALYTICAL METHOD Mahod N u m k VOOO1782 Methodof h d y a l s for the Determinitlon of P e m o ~ c u n o i Ae cid (PFOA) in Sediment by LCIMSIMS hdytlcd T&g Facility: Exygcn Rcrurch 30S8 Raurch Drive State College, PA 16801 Exygen Research Page 30 of 65 Page 96 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 -b Y P M1thDdNumbcrVOOOl181 I I AUALYnCAL METHOD Msthod of Anrly*s for the Dacrminsrionof Pduomoctanoic Acid (PFOA) in Sediment by LC/Ms/MS 1.0 scape Thii m d u 10 k employed for the iuolstionand qvPIltitationof ~ u o m o c t ~ o i c acid by Hi& Puf&mance Liquid chmrmtography coupled to a tandem Mars SpcetromchicLkktor (LcIMS/MS)m acdimmt. 2.0 safety 2.1 ruwsy8 obrsnc d e lrboruory practicer. 2.2 C o d t thc appmpaialcMSDS before handling any chemical for proper safety prrcurtoIu. 3.0 SatnpkRcquimmt 3.1 AtI~30goftutumplcforcrmstion. 3.2 No umple procsuinp is Medsdfor d m m t umplu. 3.3 Sunpla atorcd m5igcratcd ahodd be allowed to equilibrate to room 1CUlpWWS. 3.4 All umplm musibe thoroughly mixed before being umpled for extraction 3.5 Sample collectionpmccdurm will be rpsificd in the sampling plan for this 4.0 RupaumdSt.ndudr - 4.1 Wacr HPU:grade 4.2 McUlnnol-HPLcpdc 4.3 Acetic Acid - Rupcnt pdc - 4.4 Ammonium Accutc A.C.S. R q a u Gndc - 4.5 Pcrflwrooctmaic Acid Sigma-Aldrich 5.0 ~ n r t r u w s l l d E q u i p w n t 5.1 A high pcrfarmncc liquid chmmuograph capable of pumping up to 2 rolvcnu equippoi with L vuiable volumc injector capable of injecting 5-200 pL coonootcdto a undm M u c Spcnromctcr(Lc/MS/MS). 5.2 A device to collect raw data for peak integrationand quantiution. 5.3 Anrlyliulb a l m capableof d i g (0O.oooO1 g. 5.4 50 mL dirpoublepolypropylmccentrifugea b . 5.5 15 mL dirpoublepoljpmpylmcccllailuge a b . 5.6 DirpoublemiMpipoU (501oouL, 100-200uL). 5.7 12s-mL LDPE numw-mouth bottlm. 5.8 2 mLc l w HPLC vial kit. 5.9 D*poublopipctta. 5.10 Autopipeam(lWlooO p L u d 10100 pL), with ditposobletips. 5.11 W.tsnSopP~V~.c6cc(Ip)tClBSPEcu(ridga. 5.12 SPEvruvmmrtrjfold. Page 31 of 65 Exygen Research Page 97 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 ,- I I ANALyllCAL METHOD Mnhcd of Arulytit for IhcDC(armnr0on of PedwmofunOic Acid (PFOA)in Sediment b\ 5.13 Vomer. 5.14 W r i ~ l - . ~bt ik~ . 5.15 Cdri%ge capable of @innin#SO mL polypropylene tuba at 3000rpm. 6.0 cbromltopnphicsptcm 6.1 W ~ UCollumn: FlwphwRP (Keyrtonc Scientific).2. I mm x 50 mm. 5p ('M82505-052130) 6.2 Tanpaauc:3o.C 6.3 Mobilo phvo (A) : 2 mMAmmonilrmAcetate in Water 6.4 Mobile Phuo (B): Methanol 6.5 WonIRogram: Timc[miolu 0.0 65 I .o 65 8.0 25 20.0 25 22.5 65 Flow Rate ULmlLminl 35 0.3 35 0.3 75 0.3 75 0.3 35 0.3 - 6.6 hjjostionVolumc: 15 pL (uabe incrcuedto Mmuch u SO pL). 6.7 Qudtuion: PaL A m -exlanul standud cdibruion CWC. 6.8 Run Tim: 23 minuter The obove waditi~lvM inlcodcdM P guide and m a y be changed in order to optimiro chc WLC syttcm. 7.0 MslMs Syrlan 7.1 Mode: ElsWQnyNOgntiVCMRMmodc,monitoring413 -b 3 6 9 d Z for PFOA. Tbs obove wnditi~lvM W e d u a guide md may be changed in order 10 optimirctbsMsMstydan 8.0 mmto inof SolUIionr 8.1 Mobile phvc 8.1.1 2 mM uauonium w(.(c in wucr is prepared by adding 0.154 g of unmnium &e lo 1000mL of water. Exygen Research Page 32 of 4S Page 98 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 I 1 ANALYTICAL METHOD Mabod of AtuIvlis Cthe D*omuuu'on ofPaflwroocuMic Acid (PFOA) in Sedimentb) LC/MS/MS 8.2 Exovtion Solutiolu 8.2.1 1% .caiCloid in water u prrprredby adding IO mL of acetic acid to IMX) mLofw.tar. A l m . t c volumorm y bo prcpusd. 9.0 StradvdRcpurtion 9.1 Stlldvd Stock/Fortification Solution 9.1.1 Rspuca stock wlutiw of -I00 p#mL of PFOA by weighing IO my, of uulylicrluvldvd (corned for purity) and dilute to 100mL with moth001 in a IZS-mL LDPEbottle. 9.12 A IO p#mL fortificationrolution of PFOA is prepared by bringing IO mL of tbo 100 pghnL solution lo 1 bl volume of I 0 0 with mcrhinol in a I25 mL LDPE bottle. 9.1.3 A 1.0p&L fortifidon wlution ofPFOA is prepared by bringing IO mL of the 10 p#mL wlulion to a final volume of 100 with mcthrnol in a 125 mL LDPE bottle. 9.1.4 A 0.1 p@ fortificationlolution of PFOA is preparedby bringing IO mL of the 1.0 p#mL colution to a hod volume of 100 with methanol in a125mLLDPEbo(tlc. 9.1.5 A 0.01 p@nL fortifiutionMIU~~OIIof PFOA is prepared by bringing 10 mL of the 0.1 p#mL wlution to a final volume of 100 with mdunol in a I25 mL LDPE bottle. 9.1.6 The mck md fortificationlolutionr art to be slorcd in a refrigeratorat lppmxinutoly 4C d M rtable for a muimum period of 6 months fmm tbo date of pmparation. 9.2 StltldudC.LibntionSolutionr 9.2.1 L#MSihlS ulibntion atudlrdr UT p r o p a d in mcthanol via dilution oftbo0.l p#mL fortificationwlution. 9.2.2 TICfollowing is 1 lypicd a m p l e : additional concentralionsmay be PTCPUCdUIIcodsd. cmcm- Find ofFoniliution Volwnc Dilutcdlo Concmmtion Solutim(nplmL) (mL) (mL) (np/mL) I00 10 100 10.0 100 5 100 5 .o 100 2 100 2.0 IO IO I00 I .o 5 IO IM) 0.5 2 IO 100 0.2 Page 33 of 65 Exygen Research Page 99 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 .. I I ANALYTICAL METHOD Muhod of AIyIpia forb D*sminUionof PerfluorwcUnoic Acid (PFOA) in Sediment by Lmms 9.2.3 Stom d UlibRtlon aundudr in 125-mLLDPE nnrrow-mouth bolller u 2'C to 6.C. up IOsix m o n k 9.2.4 Altmutc voluma urd concmmtionr of a l p n d u b may be prepared os nocdcd. 10.0 BucbSctUp 10.1 Euh batch of vnqrla cx(Trlcd (typiollly 20 or Its) musl include 81 ICBII one conrrol md two untrorted cootrolr fortified ai known coIwlPDDoM(lab conbul Qiko) to vmfy procedural recovery for the batch 10.2 Rsqumnmufor deld and hborato~d' uplicaica and spikes will bo rpcsilicd in h e q d t y UWUKX p h for h r project 11 0 SvnplcExtncpon 11.1 11.2 11.3 11.4 11.5 11.6 11.7 11.8 11.9 11.10 11.1 I 11.12 11.13 Weigh 5 g of llmplc into 50 mL polypropylae centrifuge lubes (fomfy as needed, rcplw lid ud mix well). Add 35 mL of 1% pcaic acid, cap, v o I t u and &&e on a wrist action sbnker for do minuta. Ccntrifugothe Nbaat -3wO rpm for -20 minuter. Condition tbc CISSPE Cprtridgea (1 g, 6 mL) by passing 10 mL methanol followedby 20 mL ofHPLCwaIw (- 2 dmp/scc). Do not let column run dry Load (decant) thc umplo on h e conditioned C,I SPE canridge. Dircurd elwe. Add 20 mL of mah.nol to the tcdimat left in Le bottom of the 50 mL mtrifuge d e . Cap, vortcx and duke on a wriat action &&cr for -30 minu-. 6 c Catrifuno thahlbar u -3w0 mmfor -20 minuter. Dcunt maluno1onto the &me SPE cartridge. Collect tho eluate. W u b the solvrrm with 4 mL of mS(ha0l. CoUeclthe eluate and add it to the eluUe collalod in Ucp 1I .8. Conditiona second CIISPEclrrridps (I g, 6 mL)by passing 10mL methanol followedby 20 mLof HPLCwater (- 2 dmp/scc). Do DO let Icolumn run dry Add thc mc4bu10l to -200 mL of water and load on the secund conditioned SPE cutridge. Elute wilh -5 mL loof( mahrwl. Collecl 5 mL of CIUIC into graduaied I5 mL polypmpylmomttifligo t v h (final volume = 5 mL). Aualyro umpla wing claburpny LC/htSIMS. Exygen Research Page 34 of 63 . __ Page 100 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 I I ANALYTICAL METHOD Msthod of M y a i s for tho Dozamidon of Pcrfluorooctonoic Acid (PFOA) in Sediment by LCIMs/MS 12.1 Iqject tho U ~ m C ount of csch stmd.rd. m p l c ud fonificd sample into the LUMSMS system. A ulibntion at.nd.rd muat precede and follow all d y r c d :ampla. 12.2 St.ndudr of PFOA cotrrrpondineto at leut five or more concentration ICVC~E must be included in an andyiiul ICI. 12.3 An entire Iot of oxtncted calibration itandud8 muat be included at the beghhlg md U ths ad of a Snmplc rel. %ndardl must be interspersed brawart every 5-10 umplu. A8 an dtemativc, an entire set of calibration r(nadudr may bc injected at the beginning of a set followed by calibrahon ctaduda i M c n p d way 5-10 umpla (to account for n second set of r(mdudr). In either UIC, ulibration rtmdarda must be the fim and last injection in ample Iot. 12.4 Uw liacu ltpldvd c w a for qumtitetion. Linear standard curvu are p a r t e d hntho vulytc by linear ragmuion using I/x weighting of peak area v w y l d b d o n atandud conccntntion usin8 MsrrLynx 3.1 (or cquivaleni) lDfhvMa m . 12.5 Smple mspine lhould not ex& ltudud ruporua. Any samples that ox& stmdatdrorporuwlhould be furtherdiluted and mandyred. 13.0 AccoptracoMtosir 13.1 c h r o r m t o ~must rbow I peak of r dauphter ion .t 369 MU hom a parent of 413 ma Ibc 413 m u pvcnt ~orrclpondrlo the PFOA anion, while Ihe daughterion (369mu) npmcDtr tho IOU of cubondioxide. 13.2 Method blrnlrr must not Ooncrin PFOA at lwelr groatcr than the LOQ. If a blank contain8 PFOA It levelt greater than 0.2 W L . then a new blank umple mult ba obtainedmd tho entireact must be rc-cxtnctcd. 13.3 Rocovosia of contml lpikcl d mahix spika must be between 70-130% of their lmDm vrl~ar.If a control spike fall1 outaidc the lcccpfablc limits, the e O h ad of umpla Ibmld bc raabacied. Any matrix spike outside 70130% lhould k wduated by the d F t to d e t m i n e if re-extraction is wunntod 13.4 Any dibfuim atandud found to bc a atatiUid outlier by using the Huge Error Tort.MY be oxcludcd from thc c d d U i o n of the calibration curve Howova. Ibo told number of oxtnctd calibration s u n d u d s tbat could be oxc~udcdmust not uccsd 2C% of the total number of cxmcted standards injected. 13.5 The cormlUion Ewffiucnt (R) for calibration SWCI generated must be 20.992 (R' 20.985). If catibnuion ruulta fall outside thue limits. then sppmpriltc ltcpr must be taken to d j w t in~ttumentoperation. and the st.ndudr or thc rclevant set of ramplo rbould be reanalyzed. Page 35of 65 Exygen Research Page 101 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygcn ProtocolNumber:PO000760 1 1 ww- McrtbodNumber V00017Bl ANALYTICAL METHOD Mcthod of Andpis for tho D e t a m i ~ toif~P4~u~omocmoic Acid (PFOA) in Scdimcni by LCJMS/MS 13.6 Rotontion timar bchvoan s u n d u b and ramplea must not ~ f miore than f 4 94 within an uulylicd nm. IfmentionrimCdriR exfwdr this limit within an uulyticdrnthm the let mustbo d y z c d . 14.0 Cplsvlatiom 14.1 Use thc following equationto cllculatcL e mount of PFOA found (in ng/mL. b w d on peak uu)wing thc tUndvd curve (linear regression paramaersl gcndcd by ths Maw Lynx M ~ U Wp m p m : P F O A f o u n d ( @ m L ) - ~ ' . '1 x DF DF - factor by which the final volume w dope u diluted, if nccuury. 14.2 For umpla f d d c d with known ~ o u n t aof PFOA prior lo cxwtion. use - tbo following equation to cdculale theporccnt recovery. R k o v q (%) - 14.3 Use m0 followingequationto c o n v a the unounlof PFOA found in n d d to ndg W). F'FOA found @pa) x firul volume I S umplc weigh1(5 e) 14.4 U#c tbe following equation (if IIOMUY~)to calculate L e mount of PFOA found in ppb bwd PFOA found (ppb) dry - on dry weight. weight PFOA found @pb) x [ I WK / total rolidr(%)l P.p.1 of I Page 36 of 65 Exygen Research Page 102 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 ANALYTICAL METHOD __ Molhod N u m k V0001783 Method o f h d y s l c for the Detennlnatlon of Perlltloroc-rtmolc Acid (PFOA) in Fish and Clam by LCIMSIMS AnalyticalTcsting Facility: Exygen Research 3058 R-h Drive State College,PA 16801 Approved By: TechnicalLcdcr.LC-MS.E x ~ R~sQlsMh IPIl.bfPJ Date Exygen Research Page 37of 65 -- Page 103 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 E x m llnsuEh M r W N k r VWOl783 I I ANALLYTICALMETHOD Mcchod of Aorlpir for the DO(crminr1Mnof PcttluMooct.noicAcid (PFOA) in Fish and Clam by Lc/Ms/MS 1.0 scope Thir mclhod is to be amploycd for the iwlaiion ud gumtitation of perfluomtanoic r i d by High Perfommace Liquid Cluonutognphy coupled lo a landcm Mass Spcctmmchic Detector( L c / M s N S ) in fuh ud clams. 2.0 safely 2.1 A l w i p obwve safe Ilboralo~p~rssticu. 2.2 COMUIIthe rppmprialc MSDS before handling any chemical for proper rnlbly pIzcpuIions. 3.0 SvnpleRcqvirrmCnt 3.1 Atl~20pofrcnlvaplcforwrhaction. 3.2 Suapla lbovld bo p d bofore ahaction. Place thc frozen sample in a food proccuor md bmopcnizewith dry ice. Place the samples in containen aud leave opm in l?ozen storage overnight to allow for cnrban dioxide sublinu!ica Sed and plaw the implu in frozen storage until lime of dylis. 3.3 Sunple collcetion p d m will bo lpscihcd in the sampling plan for this pmjcct. 4.0 R u g c o t r m d S ~ 4.1 Wales-HPLC grade 4.2 Acctonibilc-HPLC grade 4.4 4.5 4.6 4.1 4.8 4.9 4.10 4.11 4.12 4.13 5.0 Iruhumclll md Equipmoat 5.1 A high pafomun~li~quid chmnutognph capable of pumping up to 2 wlvau equipped with a variable volume injector clpablc of injecting 5-2W FL coonsrsdIo P tandem MuSpecoomuu(LCMSMS). $ 2 A device to mllcclraw b fororpclk integration and qumtitalion. 5.3 Aorlyticd bllrncc capable of reding to O.oooO1 g. PIgr 2 "f 8 . . _._ . Page 38 of 6s - .- Exygen Research Page 104 of 140 ~~~ ~ Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 Method of Anal+ for the Daaminuionof PcrflwmoclanoicAcid (PFOA) in Fish and Clam by Lc/MsIMS 5.4 R o t . r y o v l p o ~ r . 5.5 TiuUmira. 5.6 I25 mL pur-rhrpod h l u . 5.7 50 mL dilpoublcpolypmpylcnc ccnhifuge tube:. 5.8 I5 mLdilpollblepolypropylmecmhifugetubcr. 5.9 Dirporrblcmicmpipda(50-100uL. IW-ZOO&). 5.10 125mL LDPEnurow-moutbboctlcc. 5.11 2 mL clur WU:vial kit. 5.12 Diapoublc pipottcr. 5.13 A u I o p ~ ( 1 C # l 0 0 0 f l L ~ dlDlOOflL),withdirpoublelipr. 5.14 SPE tuba ( 2 W ) (Supclco ut.no. N057177). 5.I5 Wrist action rh.Lu. 5.16 ConbifLoccapable of Ipinnhg 50 mL p l ~ m p y l e n eNbuat Zoo0rpm. 6.0 chromstopphic Synm 6.1 Analytiul Column:Fluophuc Rp (KeynoneScientific),2.1 nun x 50 mm. $I (PN:82505-052130) 6.2 Tarpcnhm: 30.C 6.3 Mobile Phve (A) : 2 mM Ammonium Acetate in W . t a 6.4 MobilcPhw(B): M h l 6.5 GndimtProgun: rimoou 0.0 65 I .o 65 8.0 25 20.0 25 22.5 65 Flow Rue YULmlLminl 35 0.3 35 0.3 75 0.3 75 0.3 35 0.3 - 6.6 InjectionVolumo: IS pL (wbe inacucd Io M much M 50 FL). 6.7 Qumtitation: P u k Area- cxtund 1Undrrdcalibrationcurve. 6.8 Run Time: 23 minUta. The dmve condition,uo intended u Ifide and may be changed in order Io optimizc thc HPLC syltsm. Exygen Research Page 39 of 65 Page 105 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 1 ' exygcnRuuxh MctbodN&z VooO1781 ANALYTICAL METHOD Methodof U y r * fm tha -on of Perflvoroocunoic Acid (PFOA) in Fish and Clam bv LcIMs/MS 7.0 MSlMSSyrtsm 7.1 Mode:Eloctrospny NepUivc MRMmode,monitoring413 +369 m/z for PFOA. The h v e eoaditiorvM intcndcd u I fide and m y be changed in order IO op(imizethe MSMS syltan. 8.0 prrP.ntion of Solutionr 8.1 MobilePhvo 8.1.1 2 mM uurnonium .oartc in wola is p q d by adding 0.154 g of dum-to 1WOmLofw.tUr. 8.2 E x d o n Solutinm 8.2. I 2% ucorbic acid in mc(hm0lu prepared by diuolving 2 8 of ascorbic acid in I 0 0 mL ofmcthol. 8.2.2 30% D ~ y l d i c b m s i l a n ien Iolucnc is prepared by bringing 3 mL of diutathyldichlormil.aeto a firul volume of IO mL with toluene. Altermtc volvmcrm y be prrp.rod. 9.0 suaanrdwon 9.I Strabrd S W o r t i f i u t i o nSolution 9.1.1 9.1.2 9.1.3 9.1.4 9.1.5 RcpMn atock solutionof -100 pphnL of PFOA by weighing 10 mg of OJYtiUl suaanrd (wmtcd for purity) and dilute to 100 mL with m t h ~in~al125-mL LDPE bottle. A 1.0 pdmL for(i6catioowlution of PFOA is prepared by bnnging I mL of the 100 p g h L solution to n I i ~ vlolume of 100 with methanol in n 125mL LDPE bott~e. A 0.1 pghL fortificuion wlution of PFOA is prepared by bringing IO mLofthe 1.0 cdmL wlution to a fmd volume of 100 with me~hanolin a 125 mL LDPE bottle. A 0.01 pdmL fortification wlution of PFOA is prepared by bringing IO mL of the 0.1 pdmL wlution to a final volume of 100 with nwluwlin a I25mL LDPE bottle. The at& md fortification wlutiorv am to bs stored in a refrigerator at qpnxinut8ly 4'C and uc slablc for a maximum period of 6 months fmm the dateof prcpmtion. Page 40 of 65 Exygen Research Page 106 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygcn Protocol Number: PO000760 -Exyam Mebod Number V0001783 1 ANALkTICAL METHOD Metbodof Atulyds601 thc Ddaminationof PerfluorooctPnoicAcid (PFOA)in Fish and Clam by LC/MS/MS 9.2 Slaadard Calibration SOlUtiOM 9.2.1 Lc/Ms/MS calibrationNnd.rdr arc preparedin methanol via dilution ofthc 1.0 p&L forlificltion wlution. 9.2.2 The following is typical exnmplc:additionalc o ~ c e n ~ a ~ m~ aoynbse propped u nceded. conccau F,d ofFarifiutim V d w Dilutedm Cmcenmtion SOlutiaolllmL) (mL) (mL) (m/mL) 1.0 5.0 100 0.05 1.0 2.1 100 0.025 1.0 1.0 100 0.01 0.05 IO 100 0.005 0.025 10 100 0.0021 0. I IO 100 0.005 IO loo 0.00I 0.m5 9.2.3 Store dl Wlibntion 8lduda in 12SmL LDPE runowmouth bottles .t2% to 6% up to nix moatha. 9.2.4 VO~UmWud WnCUlklIiOMOf S l a a d d S may bC PrCpPrcd 15 Eecded. 10.0 BuehSnUp 10.1 Eub bat& of umplu nrtnsIed(lypicdly 20 or less) must include at least one unbwed wnhul and two untrattd contmla fortified at known wncenmtioru(I& wnaul @&E) to veri@ pmocdunl rrcovcry for the batch. 10.2 Rcquiranmu for Beld and labomtory duplicatu and spikes will be specified in thc quality UNMCC plan for thir project. 11.0 SampleExtraction 11.1 Weigh 5 g of k o m umple into SO mL polypropylene centrifuge lubes (forti@uneeded. rcpllcclid andmix wen). 11.2 Add 30 mL of lsctonibilemd &&e on a m i s t action ah&.% for -1 5 minutes. 11.3 Place thc tuba in IKwza for -I hour. 11.4 Pack ud cwditiontbc SPENbuud ailvlizc the pcar-rhpped flasks. 11.5 Pack the 20 mL SPE lubw in sequence with 2 g florid. 2 g silica gel. 2 g cubon, and 1 8 LC-NH,. Condition the column8 wilh 20 mL of mahanol. tbcn 20 mL of acdonidlc. Dincud dl w u h a . Do not allow the column to dry. 11.6 Silsnisc the I25 mL pur-lb.psd flplkr by rinsing with the 30% dimdhyldichlorwih in t o h wlution. Rinse thc flaskwith tolueneonce, followed by mchnol (btimu). Dry the fl& wmplclcly before use. eilha by &drying 01 wilh a -1 of nitmgm. Page 41 of 65 Exygen Research Page 107 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 Expltacach I Mcthod of Analpis forthc ANALYTICAL METHOD 1 ~ a m ~ ~ l l l b e r v m i i a ~ of P n f l w ~ t s n o i cAcid (PFOA) in Fish and Clam by LC/MS/MS 11.7 11.8 11.9 11.10 11.11 11.12 11.13 11.14 11.15 11.16 11.17 1I. I8 Centrifugethe 50 mL p o l y p ~ y l m teube0 containing rnmple at -2000 rpm for-lOmiWtsl. DCCMtIhe cxhrt on to a wnditionod SPE column fitted inside the mouth 01' the par-rh.psd flask. CoUscl the clualc in the 125 mL silanized pear-shape flask. Add IO mL of ICa(0nitrilc Io the umple in the 50 mL centrifuge tube. Homogoniw the frozen fat phase wing a IiMumiza for -30 sccondr and rinse the tiuvmircrwith -10 mL of .Cnonitrilc into thc tube. ShLctbournplc~~for-lOminutclonamiat-ffitionrhnka P b t h c h t b s r i n a f h - I hourmore. cclleihgatbo 50 mL polypmpylme ~ b caontainingumplc at -2000 rpm for-lominutea. Deunt thc exlnct onto the m e SPE wlumn. Collect the eluate into the rune peu-llupcd fluk ud w m b k with the eluent from the initial cX&W. Pur 20mL of acetoNbilc lhroughthe SPE column and combine the eluate in thc ume pear-duped flask. Add 3-4 drops of I-octulol to the cxlracl in the pewshaped flask and evaporite u reduced p m wing a mtuyevaporator(01z. 40'C). M8kc the tirul volume. by adding 2 mL of 2% ascorbic acid in methanol to thcpar-ih.pcdfluk ud swirl to mix/diuolvc. Tnnafu the cxmcb to HPLC vida wing dirpoaablcpipcu. Analyze u m p l a wingelecbuspray LC/MS/MS. 12.1 wect the u m e llwylllof each rundud. umplc and fortified sample into the Lc/Ms/MS rptan. A u l i b d o n standard mwt precede and follow all d y r e d wrplol. 12.2 Sunda& of PFOA wnwpondinglo It lwt five or more concentrationlevels muat be included in m aulyiicd act. 12.3 An enth acZ of calibration aunda& mu1 be included a1 the beginning and at the a d of 8 umplc act. Sundvdr mwt be intcnpcraed between every 5-1U umpla. At an dlunativc. m entire act of calibration standards may be i n j d at the beginning of a set followed by calibration standards inmapaned wuy 5-10 lamplei (to .ccount for a iccond Mof standard:). In citha case. ulibntion standub mwt be the first ud last injection in a umple IOL 12.4 Uac linear d.rdud ma for qtuntitstion. L i n w standard curves are pcntcd forthc d y t c by linur regrearionwing I/x weightingof puk area vumu cllibntion atandud wnccimation wing MurLynx 3.3 (or equivalent) mflwm ayltan Page 42 of 65 Exygen Research Page 108 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 ww- Metbod Number VooO1713 1 I ANALYTICALMETHOD Mothod of Andpi#for theDetcrmhUbn of PerfluorwcIanoic Acid (PFOA)in Fish and C l a m bv Lc/Ms/MS 12.5 Sample nrposw should not exceed Nndnrd ruponrsr. Any samples that exceed sundud rclponresshould bc m e r diluted and reanalped. 13.0 Accqmcscnfai. 13.1 chmnutogrmmust rbow I peak of a daughter ion at 369 m u from a pnrcni of 413 m u . Tho 413 m u p m t wncspondr to the PFOA anion. while Ihe daughter ion (369 mu) rsprotcnuthe IOUof carbon dioxide. 13.2 Mahod bl9*c mvnf no1 wntsin PFOA at lcvclc p a l e r lh8n the LOQ If a blank Eontaitu PPOA at levels p a l e r than 0.5 ppb. then a new blank sample mvnf bo obtained and the a~tircret must be rc-cxlruted. 13.3 Rsoovaicr of control spiku and matrix qiku must bc bcovcen 70-1 30% of thdr known valua. If I w n m l spike Wl. outtide the acceptable limits. the clltire ICz of umplol thould be re-oxh.GtOd. 13.4 Any ulibntion mandard found to be a atatistiul outlier by wing the Huge Error Tort,m y be excluded lrom the calculation of the calibration CUNC. HOWCVKt.hc total numba of u l i b d o n strndrrdr that could be excluded mvnfnot axwd 20% of thc totalnumberof rund.rdr injected. 13.5 shc m l . l i o n coefficient (R) for calibration curves generated must be 20.992 (R' 20.985). If calibntion ruultr WI outside thew limits, then Ippmpriat~s k p ~muat bc taken to adjust instrument operation. and the suadudc w mC mlevant act of tunplushould be nmalyzcd. 13.6 RMltion rime bawm c w d u d r md samples must not drill more than f 4 K within m uulylicd run. Ifretention time drift cxcccds this limil within an d y t i u l tun then the ret must be runrlylsd. 14.0 Ca!alatioru 14.1 UICthc foUowin#e q d o n to clkul.tc the amount of PFOA found (in ng/mL. b a d on peak uu)using thc atandad curve (linear r e p s i o n paramctcn) - g~lsntodby tho M u r Lynx K&WM PFOA found (ng/mL) pmgnm: dope 14.2 UICthc following equationto wnvat thc mount of PFOA found in ng/mL lo nd8 (PPb). - PFOA found @pb) volume (mL) x DF] sample weight (p) -DF factorby which the fuul volumew u diluted, if necessary. Page 43 of 65 Exygen Research Page 109 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 Exygcn RewucLI M e W Number V0001781 1 I AN.4LYIIC.U MF,THOD Method of Aarlflir forthe DacrminuionofPcrtlwmlMoic Acid (PFOA) in Fish and Clam by l.CMS/MS 14.3 For umplw fOrti6cd with known unounui of PFOA prior to extraction. use R e - the following covey (%) equation (0 c l l c u l a t e the percent r c c o v n y . Exygen Research Page 44 of65 Page 110 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 ANALYTICAL METHOD McUlodNumk V0001784 Metbodo l h i l y h lor the Dctuml.ition of PsrfluoroocUnoicAcid (PFOA) in VcgcUtlon by LCMSrmS Exyga Rucsrch 3058 R w u c h Drive St.cs Collspc. P A 16801 Approved By: Exygen Research TotalPagu: 7 Page 45 of 65 Page 111 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 Exygen Rnoush ~erbo~~vmbcrvw17a~ I A%lLLFTICAl MhTROD Method of M y s i c for the Determimionof P d w m o e m o i c Acid (PFOA)in Vegetation' by L C M S M S 1.0 scope This mdwd is to be amploycd for h e iwluion and quantiruion of pcrfluorooctanotc acid by High Pcrformrncc Liquid chmmatography coupled to P mdcm Mars SpcctromekicDetector (Lc/MS/MS) in vegetation. 2.0 safely 2.1 Ahvayco b i w e UTc lpbontoty practicss. 2.2 Conauk tho rppropri.lc MSDS before hlndling MYchemical for proper safely prec.utiolu. 3.0 SlmplcRequimnat 3.1 At leut 20 g of Mumplc for exmstion 32 Svnpla lhould k pracucdbefore extraction. Place the frozen sample in a foodpmwmr ud homogak with dry ice. Place the m p l c s in containcs ud lmve opcn in frozcD #torage ovcmipht to allow for carbon dioxide s u b l i i Sed d p l w thc lnmpl~sin frozen ston~cuntil tune of uulysis. 3.3 Sunplc collection p d u w will be specified in ulc sampling plan for this project. 4.0 RagatsudSundvdr 4.1 W.la-HPLCgrde - 4.2 Acetonikih HPLC & - 4.3 Cubon(120-400 mwh) Reagent grade 4.4 M c l h m O l - H P U : ~ - 4.5 Silicagel (60-200 mesh) Reagent @e 4.6 Floriril(aO-l00 mash) - R a g a t @e 4.7 Supaclun LC-NHz - Reagent @e 4.8 I-OctlaOl- HPU: pndc - 4.9 L h r b i e r i d Reagentgndc - 4.10 Dimstby!diobhm*lmo wail grade 4.11 Tolucoc-Rugentpndc 4.12 Acwc-A.C.S. GRdc 4.13 Puflwmocuwio Acid - Sigrm-Aldricb 5.0 ~ d E q v p m s n t 5.1 A high Paapmunsc liquid chromatograph capable of pumping up to 2 tolventc equipped with &able volume injector capable of injecting 5-200 pL coMc*odto a ludanM u l spcchom*cr (LCIMSMS). 5.2 A dcviwto collect nw data for p u k integrationand quantitntion 5.3 Adytical b a l m capable of Mding to O.oooO1 g. Page 46 of 65 Exygen Research Page 112 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.:PO000760 Exygen Protocol Number: PO000760 ExymRru~sh Mnbod Numbcr VOW1784 muyric,u WTHOD 'Mehod of M y i a for the Dctamirution of Pduomoetlnoic Acid (PFOA) in Vegetation by LCMS/MS 5.4 Rouryevlpontor. 5.5 125mLpur-lh.pcdoMka. 5.6 50 mL dilpovblo polypmpylcnccenltihge ma. 5.7 15 mL a*pollslc polypropylenemltifuge tube:. 5.8 Dispoublcmicmpipe~(SO-loOUr,lOO-ZWuL). mu: 5.9 125-mLD P B narrow-mouthbottle& 5.10 2 mL clear v*l kit. 5.l'l Diapoublepipoau. 5.12 AuIqipeItu(IM1000pLaud IO-1M)pL),withdisporPbletips. 5.13 SPEtuba(PCmL)(Supclcocat. no.NO57177). 5.14 Wrilt actionrh.lrpr. 5.15 Ceneihge~~leofrpinnine50mLpolypropylwKNb3ut Z W Q m . 6.0 C h m m U o ~ c S y t a m 6.1 Analytiul Column: FluophprcRP (Keystone Scientific).2. I mm x 50 mm. 511 ( P N 82505052130) 6.2 Tanpmlure: 30% 6.3 Mobile Phue (A) : 2 mM Ammonium Acetate in Water 6.4 Mobile Phue (e) : Mah.nol 6.5 GndimtPm&nm: ndlnird%A 0.0 65 1 .o 65 8.0 25 20.0 25 22.5 65 Flow Rate UhUKQw 35 0.3 35 0.3 75 0.3 75 0.3 35 0.3 6.6 6.7 - hjcctioo Volume: 15 pL (w e inaCaud to u much u 50 p[ Qulnlit8lion: Puk Arm exlanrl W a r d calibrationcurve. 6.8 Run Timc: -23 minutes. The above conditionr are intadcd u a guide and may be changed in order to Optimk tho 8yuCm. 7.0 W S S y d C m 7.1 Mode: ElcchapnyNe@ve MRM mode, monitoriq 413 + 369 d z for PFOA. ._ Exygen Research Page 47 of 65 Page 113 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 I ' F x m L m r c b MotbdNwnkr VWJOI784 AX4LYTlCirL !WIXOD Method o f h d p i s for the DUamination OfPduorooeIanoic Acid (PFOA)in Vegetation bv LC/Ms/MS Thc lbave conditiolvM intcnded as a guide and may be changed in order IO optLnic the MSMS synm. 8.0 Plcp.ntion of Solutiolu 8.1 Mobile Phuc 8.1.1 2 mM d u m .couIc in w l t a is propued by adding 0.154 g of Lmmooium m e to loo0 mL of water. 8.2 ExWtion Solutions 8.2.1 2% ucorbic acid inmahrnol u PnpMdby dissolving 2 g of ascorbic r i d in LOO mLofmclhsaol. 8.2.2 3oK ~ y l d i c h l o r o a i l ain~to~luene ir prepared by bringing 3 mL of dim&yldichlmsihe to a fid volumeof IO mL withtoluene. A l t d e volumcl may be p e p r e d 9.0 SundvdPrepmtion 9.I Sundvd Surwportifidon Solution 9.1.1 Prep= a stock solutionof -100 &mL of PFOA by weighing IO niy o f d y I i u I sUndud (correctedfor purity) md dilute 10 100 mL with mahrnol in i125-mL LDPE bottle. 9.1.2 A 1.0 (rghnL forrificlcion Iolution of PFOA is p r c p d by bringing 1 mL of the 100 pg4nL wlution to a find volume of 100 with methanol in a 125 mL LDPE bottle. 9.1.3 A 0.1 p#mL fo6fiaIion wlution ofPFOA is prepared by bringing IO mLoftbc I.Ou~~solutiont4ahlL.lvoIumeo10f 0withmethrnolin i125 mLLDPfbotlle. 9.1.4 A 0.01 pp/mL f a r t i 5 c i t h wlution of PFOA I: prepared by bnnging 10 mL of cbo 0.1 pg/mL wlution to 8 find volume of IW with mclh.nd in a 1x5 mLLDPEbotlle. 9.1.5 Th stock and fortificationwlutiona arc to be aorcd in a refigerator at yl- 4OC lad us s1.blc for imaximum period of 6 months fmm the date of preparation. 9.1 S~andudCdibntion Solutim 9.2.1 LCMSiMS d b n t i o n :Ianda& u e propared in methanol via dilution of the 1.0 pghnL fortificationwlution. Page 48 of 65 Exygen Research Page 114 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 Exyamb- M e W Number VWOIl84 I I AhlrLYIICAl. MYMOD Mcthod of Analyds for the Ddwmiartion of PafluoroOarnoic Acid (PFOA) in Vegetation byLClMWhiS 9.2.2 Tho foUowing is ;Itypical example: miditionel concmaotionr may be pqmedun#dcd. CaKaCRtioa Fuul of Fortifdon Volums Diluted to Cowsnuation Solutim~g/mL) [I&) (mL) I .o 5.0 100 I .o 1.5 100 OldmL) 0.05 0.025 1.0 1.0 100 0.01 0.05 IO I00 0.005 0.021 IO 100 0.0025 0.I IO 100 0.001 0.005 IO 100 o.ooo5 9.2.3 Store all c d W m stah& in 125-mL LDPE nurow-mouth bottler at 2'C to 6'C. up to i x months. 9.2.4 AltanUe v o l m u ud wnmaotions of slandds may be preparcd as nc+dcd. 10.0 BatchSctUp 10.1 E.ch b.tsh of umplu cxlnctcd (typically 20 or leas) must include at leas1 one unmtcd confml ud two unbuatcd wnv~lr fonificd at known coDccsltRliDau(lab control @e) to v a i w procrdwl recovery for the batch. 10.2 RcquironwnU for field ud laboratory dupliutu ud apiksr will be specified in the qualityu m c eplan for thir pmjcct. 11.0 Sunplc Exrnction 11.1 Weigh 5 g of fmzcn sample into SO mL polypropylene centrifuge tubes (fortipv M wedcd. replace lid md mix well). 11.2 Add 30 mL of rctonihile and shakeon a wriM actionshakerfor -I 5 minules. 11.3 Ccnaifupe the 50 mL polypropylmc N b u containing rmple at -2000 rpm for-lominutar. 11.4 Pack ud condition the SPE Nba and aiIpoizcthe pur-&ped flasks. 11.5 P.ck the 20 mL SPE ~ b inavqumce with 2 p florid. 2 g silica gel. 2 g urbon, ud I g LC-NHI.Condition the colurrrm with 20 mL of methanol. thsa 20 mL of raonibile. D i w d all w u h n . Do not allow the column lo ?& 11.6 the 125 mL pcrr-lbrpod fl& by rinsing with the 30% d i m e t h y l d i c h l o r o ~in~tolucoe wlution. R~MCthe flwk with toluene once. followod by mahnol (tbroo timcC). Dry the flasks~omplstclybefore use. either by airdryingor with 8 s t r u m of nitmgm. 11.7 Dcunt the abrclon to a conditionedSPE column fitted inaide the mouth of the pcrr-llupca h k . C o l l a thc elute in the 125 mL iil?.nized pear-shape tluk. Page 49 of 65 Exygen Research Page 115 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 11.8 11.9 11.10 lI.11 11.12 11.13 11.14 11.15 11.I6 Add 20 mLof mmnilrilc to tbe umplc in the 50 mLcenlrifugctube. S~thcumplo.p.iofor-IOminutaonawriit-.ctionrbPL~. Ccntrifbgc the 50 mL polypropyleac tubu contuning sample at -2000 'pm for -5 minutsr Deunt thc cxblct onto thc m e SPE columo. Collect the eluate into the mum pur-- fluL md combme with the eluent from the initial ormetioll. Rsputltcp1l.8thmugblI.ll win. Add 3 4 dmpi of I-oClmol to the cxhct in thc pc9-1haped flask and evq-an~en~reducedpmm using a mtuy evaporator(at < 40C). Makc thc Aoll vohunc, by adding 2 mL of 2% ascorbic acid in methanol to thcpsu~flukMdrnirIlom*/dirrohrc. T~ferthenrmctltoHpLcVi.lrusingdirpoaDblcpipcU. Analyze umplu wingelectrospny Lc/MS/MS. 12.1 Injw the sane mouul of ufb rundua suaplc and fortified umple into the Lc/Ms/MS sptwn. A ulibration ~tpnd.rdmuat pmcde d follow all uulyrcd umplol. 12.2 Stlndudrof PFOA comrpodqto at leut flve or mom concentration lcvcls must be included in YI uulyticd act. 12.3 An enlirc sc4 of extnctod cllibntion nandlrdr must be included at the beginning lad u the a d of a m p l c set. Exmeted standards must be intcnponcd bawocn every 5-10 sampler. AI UI dtcrnative. an entire set of oxtncted d b d m s t d a r d ~m y be injected at the bc@nning of a ret followed by armctod calibration standard8 intcnpcncd every 5-10 samples (to mu01for a vcnd of atnctcd I(uduc*). In either CMCe,xtracted calibrationd.ndudr mu81bc the fin1 and lut iqjection in a umple set. 12.4 Uw lineu Nndrrd CWCI for qrntitntion. Linur Nndard curves arc gomratsd for the d y t c by linarmolcrrim wing l h weighting ofpeak area vcmu d b d w alandudconccntntion uing MurLynx 3.3 (or quivalcnt) loftwuclyl(cm. 12.5 Sunplc rupoars &odd not c x d ruadud responses. Any sampler that a c a d Nodvdr a p o w shouldbc huthw dilutod and reanalyzed. 13.0 A c a p l ~ ~ ~ ~ C r i m i a 13.1 Ummatogrun m W i b w a PQk of a drughta ion at 369 m u from a parent of 413 mu Tbo 413 MU psrat conorpondato Ule PFOA mion, while the a.Ugtaion (369 m u ) rspmontlthe IOM of urbon dioxide. Pipe 6 017 Page 50 of 65 Exygen Research Page 116 of 140 ~~ Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.:PO000760 Exygen Protocol Number: PO000760 I 1 .ASALYTICAL ~METEOD Metbod of ADllyrirfor the Dc(aminrtionof Pafluomaunoic Acid QFOA) in Vegetation by LClMslMS 13.2 M*hod blvrlo must not contain PFOA U INSIS patcr t h th~e LCQ. If a blank d PFOA at lcvelr m a than 0.5 ppb. then a new blank sample mylt be obtained ud the entire sa mwl be rc-cxtrac.ted. 13.3 Reum'aia of cantrol apika ud rmtrix apika must be between 70- 130% of thcu knownvdw. If control apiks fdlr outside the acceptable limits. the entire sa of umpla shouldbe ro-cxoletcd. 13.4 Any ulibntion umdud found to be a at.tirticd outlicr by using the Huge Ermr Tu& m y ba axcludcd h m the ulcullcion of the calibration CUIVC. Howcver, the low numbcr of ulihtition Nndvdr that could be cxcludcd must notarced 2oK of thc total numberof Il.ndvdr injacd. 13.5 The oarcllcion mrsoiaat (R) for ulibntion curvu gcncntcd must be 20.992 (R' 20.985). If ulibntion nrults fdl outside thuc limits, then appmpriala at- mus1 bc ULOn to adjw irumunent opention. and the lImddl or thc ICkVaut I*of umplor d~ouldbo MIIpIylcd. 13.6 Raclltiw tima bawcu~ aud a m p l a must not drift more than f 4 Y.withinMdyticd run. IfretentiontimedriA exceeds this limil within UL analyticalrun then the ad must be MO.Iyld. 14.0 14.1 Uw tho fobwingequationto cdculatcthc mount ofPFOA found (in ng/mL. b w d on pclk ua)wing thc rundsrd curve (linear regression parameters) - sCncn(0dby thc M.u Lynx loftwucpmgnm: PFOA found (nglmL) rlope 14.2 Ww tho following equationto convertthc mount of PFOA found in ngmL 10 nolo 6 M . - PFOA fouud @pb) umplc weight(g) -DP f.aMby which the flual volume WM diluted. if wceasary 14.3 For runplol W e d with kunvn mount^ of PFOA prior to cx1TPct100.use - the followingequationto c Rooovcoy (40 a l w the psreoat rocovay. Exygen Research Page 51 of 65 - ___ Page 117 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen ProtocolNumber: PO000760 ANALYTICAL METHOD Mahod N u m k VOOOl785 Aaslytisd T ~ t h Fgaciliw Bxygm R c l u r e h 3058 Rwarch h i v e Stntc College. PA 16801 Approved By: Paul Coawlly I Technical Lcder, LC-MS, Exygm R d P Exygen Research ToulPqu: 7 Page 52 of63 Page 118 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygcn Protocol Number: PO000760 Exygrn Ruarch L Method of Anal* Memod Numbrr VwO1785 1 ANALYTICAL METHOD forthcDaennhtionof Pcrfluomoctanoic Acid (PFOA) in Small M M M d Liverby LC/MS/MS I .o SEope This method ii to be snploycd for the isolation and qmtitation of pertluorooctmoic acid by High Pcrfomuocc Liquid Chnmlography coupled lo a tandem Mass SpccoomCaicDokctor (Lc/Ms/Ms)in smallm ~ ~live.r. l 2.0 safely 2.1 Alwayr O b m c d e iabommypncticu. 2.2 Corvult thc wropriatc MSDS before handling any chemical for proper sarctv prec.utionr. 3.0 Sunple Rcquimnmt 3.I At leut S g of tu1 umplc for exunction. 3.2 Sunplu lhould be procoascd before ueaction. PI= the frozen sample in Q food pmocuor ud bomogcnizc with by ice. Place the sampler in containers ud leave open in from rtonge ovClni@t 10 allow for carbon dioxide rublimltion. SUI and p h thc aunplu in h m atorage until lime of d y * s . Al(Cmrt0ly. if tbac u au hUmcimt amount of sample (-less than S g), lhenM procdng u n e w u u y a$ the samplecan be ucdas supplied. 3.3 Sunple OollSaion pmccduru will be spcci6d in the sampling plan for this plojcct. 4.0 W g C l l t I m d S t d d l 4.1 Water- HPLC &e 4.2 Mcthuml-HPLCgdC 4.3 ACaonitrilc-HPLCgdC - 4.4 Ammonium A c W A.C.S. Rupcn(Gnde - 4.5 PCmuamocevloicAcid Sigma-AMrich 5.0 IrulNnMnt.ndBquipmsnt 5.1 A high liquid chmnutogmph capable of pumping up to 2 VrlvCnU opvlppedmth I vui.ble volume injectorcapable of injecting 5-200 pLconncctdtoatrndanMurSpcckome(cr(LC/MS/MS). 5.2 A &ice 10c e l l a n w d.1.for pesk integration and quantitation. S.3 Analyticalb.*ncccapable of d n g IO 0.0M)OI g. 5.4 SO mL dirpoidlo polypropylme ccneifugotubu. S.S I5 mL disposablepolypropylme ccnhifuge lubes. 5.6 Diapoaablemiuupipou (50-1OOuL 100-200uL). 5.7 125-mLLDPEnmw-mouth botllu. 5.8 2 mL c l w HPLC vial kit. Exygen Research Page 53 of 65 ____ Page 119 of 140 ~~ ~~~ Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 1 I M'ALYTICAL METHOD Mabod of Aorlydcto0th Dcmmbtioo of PcrfluomocunOicAcid (PFOA) in Smell htmnuulLivar by LCMS/MS 5.9 oirpollblepipstlar 5.10 Autopipotlu(lM)-1ooO & d 10-100pL). with disposable tips. 5.11 W 1 ~ S c p P ~ V l c 6 c c ( l g ) l C 1 8 S P E c . m i d g c r . 5.12 SPE vwuum manifold. 5.13 Tiuumiru. 5. I4 Wriit-action WR. 5.I5 Ccsltriflrgeuplble of spinningI S mL polypmpylmc tubes at 3000 rpm 6.0 ChmmatogmphicSyffom 6.1 Mylical Column: Fluoph.scRP (Keystone Scientific). 2.1 mm x 50 mm.Sp (PM: 82505-052130) 6.2 Trmpsnture: 30.C 6.3 Mobile Phrc (A) : 2 mMAmmonium Acetate in WUcr 6.4 MobilePhrc(B): Msth.nol 6.5 Gndionthgrm: F 1 .o 8.0 20.0 22.5 Flow Rate % n f i f z a h i d 35 0.3 65 35 0.3 25 75 0.3 25 75 0.3 65 35 0.3 - 6.6 InjectionV o k 15 pL (enn lxinarnred lo u much as 50 pL). 6.1 Quurtitltion: P u k Aru ntad rtladard calibration curve. 6.8 h T k -13minurs* Thc lbovc d t i o m Minladcd u a guide d my be changed in order to optimize IhoHPLC SyItom. 7.0 MWSSyrtcm 7.1 Modc: Elahuspray Negative MRM mode. monitoring413 -t 369 m k for PFOA. Thc lbove d t i o l v yo intoadd u a guide and may be changed in order to optimize the MSMS lyltsm. . Exygen Research Page 54 of65 __ -~ Page 120 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 1 1 exysa- MomDdN&r VWO 11115 ANAL\TICALXETHOD Mahod of hdyh forthc D & d ~ t oif P~effluomoctrnoicAcid (PFOA) in Small Munmrl Liver by LCIMSIMS 8.0 RcpMtion of S ~ l u t i ~ ~ 8.1 MobilePhuc 8. I , I 2 mM ammonium acelate in w.ta ir prepared by adding 0.I54 g of amnomum Lca(.te to loo0 mL of water. Altcnute volumol m y be prcpd. 9.0 SuadvdPreplntion 9.1 Suadvd StaWFonificition Solution 9.1.1 RspMirtocLsolutionof-100pghnLofPFOA byweighing IO mg of dyliul culd.rd (comted for purity) md dilute to 100 mL with mothanol in a 125-mLLDPE bottle. 9.1.2 A 1.0 p a f0rIjfiutionsolutionof PFOA is prepared by bnngmp I mL of thc IW p g h L solution to ifind volume of 100 with &no1 in a I25 mL LDPE bottle. 9.1.3 A 0.1 fo~tificationsolutionof PFOA ir prepared by bringing IO mLof thc 1.0 p a solutionto 1 dnrl volume of 100with mthanol in a 125 mL LDPE W e . 9.1.4 Thc ltoclr d foIiificatiionsolutionsMto be stored in a refrigeratorat lppmxirrvtoly4C .nd M itdlo for a maximum period of 6 months 6um the date of prapmtion. 9.2 Stlodvd Cdibntion Solutiolu 9.2.1 LCMYM.9 d b n t i o n Itlcldub Mp r e p 4 in methanol via dilution O fh0. I p g h L foltifiC&On WlUtiM. 9.2.2 The r0UoWin.gia a typical cxunple: dditional concentrations may be prrpvsdvncedcd. cancalmrim Find of Faiifmh Volume Dilulcd lo Concentration SoluhaI(nlhnL) (mL) (mL) (np/mLI 100 5.0 100 5.0 100 2.0 100 20 100 I .o 100 10 5.0 IO 100 03 2.0 IO 100 0.2 I .o IO 100 0.I 9.2.3 Sbm d d b n t i o n 8Undardn in 12s-mL LDPE m w - m o u l h bottles 812C to 6.C. up to dx months. 9.2.4 Altanm volumu and concentnlioru of iundnrdr may be prepared 01 nocded Exygen Research - . . . Page 55 of 65 Page 121 of 140 ~ ~~ ~~ ~~~ ~~~~~ ~ Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 Exypm R.uush M U NvmbrrVOOO178J 1 1 AXUYTICAL METHOD Mathod of M y d s forthe Detamkdon ofParfluomoctlnoic Acid [PFOA) in Small Munmnl Liver by Lc/MS/MS 10.0 Batch Set Up 10.1 Each bUcb of samples extracted (typically 20 or less) must include at leas1 ON uutrutcd wntml and two untruted controls fortified at known cmcenlratim(labwntml apike)to vaify pmcedunl rccovny for thc batch. 10.2 Rcquimn~~fobr field and Inborntory duplicntes and spikes will be specified in lhc quality YIUMCC plan for thispmjst. 11.0 SunpleEXmction mc - 11.1 Wagh 1 g Of umple into n 50 mL polypropyleneccnhifvgctubes (fortlfy as needed, rcplvc lid d mix well). Note hat dtcrnate weights of livcr may bc msL(uTed dspsnainson tbc umple rim available for use. I I 2 Add w m t o umple for 8 . W volumeof 10mL. 11.3 n o m o w vmpie wing a for -1 minute. 11.4 T d m 1 mL of the sample using 8 dilporablc pipette into I IS mL dkpouble Esotrifilge tubc. 11.5 Add 5 mL of lcctonitrileud rhrkc for -20 minuteson n wrist-%lion shaker 11.6 Centrifugetbc tubuu -3000 rpm for -5 minuto. 11.7 D d the supumbnt into a 50 mL. disposable centrifuge tube and ndd 35 mL of WUCI. 11.8 Condition tbc CIISPE d g u (I g, 6 mL) by paring IO mL mclhnnol followedby 5 mLof WLC water (- 2 drop'icc). Do MI let column run dry -i - 11.9 Load tho uwle on wndihoncd C MSPE c.rtridge. Discard eluate 11.10 Eluts with mL of mdbmol. Collsst 2 mL of eluate into a gruluatcd I5 mL polypmpylcaccentrifuge tube (find volume 2 mL). 11.1I A d p o umplcr wingclectmrpny m m s . 12.0 chronut0gni)hy 12.1 h j s t the ume amount ofeach Utmdud. sample and fortified m p l c into the LCIMSIMS tyrtem. A calibration rt.ndud must precede and follow all d y d s.mpla. 12.2 S t d u d s of PFOA mmpndt'ng to U lcut five or more concentrationlevels must be incMedin an d y t i c d set. 12.3 An entiro #elof ulibntion ~Lmdardsmwt be included al the beginning and ai the cnd of a umple set. Standardsmust be interspersed between every 5-10 umpla. As an d t d v c , an entire set of calibration standard6 may be injected U lhc kpming of a set followed by cslibrntion standards inrarparod way 5.10 u m p l u (to m u n t for n second set of standards) In cilhu cue. calibration lundsrds mwt be the fmt and lut injection in il umplc Is(. 12.4 UM liacu ltudvd CWCI for quuttitation. Lincar rhndard curycs are gencntodfor the uulyte by lincu regreuion wing l h weightingof pcnk area Page 56 of 65 Exygen Research Page 122 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 1 I m u m c u METHOD Method of h d y d s far the Demmhuionof P d u o m o c w o i c Acid (PFOA) in Small MimmslLiver by KMSIMS vrmu ulibntionuudrrd wncentntionuing MpuLynx 3.3 (or equivalent) cofnnmspm. 12.5 Sunple rocpoluo lhould not exceed 6tMd.rd rerpnses. Any samples rhak cxcocd nmdpd rapoares should bc further diluted and m a l y z s d . 13.0 Accspluw Criteria 13.1 Chromuopun mvrt &ow a p k of a daughter ion at369 m u from a parent of 413 mu. 'The 413 a m u parent wrreapondr l o the PFOA anion, while !he daughter ion (369 MU) rcpnrmu ths loss of carbon dioxide. 13.2 Mctbod blanks muat MI cOnt.in PFOA at level6 greater than the LOQ. If a blank wntaiau PFOA .Ilevels p t c x U.n IO nd&then a new blank sample must be obtained and Ihcen& M1 mud be re-cxkactd. 13.3 Rocovaia of conhol spikes md matrix lpika muat bc betwccn 70-130% of their knownvalua. Ifa concml spike falb ouuido the accepublc limits. the aotirs IU of umplor h u I d be rr-olhs*sd. Any mMix spike outside 70130% lhwld be evaluated by the vulylt to dctmnine if re-extrsction is WvMtcd 13.4 Any ulibntion uudrrd found to be a rt.tidcal outlier by wing Ihe Huge Ermr Teat, m y be excluded fmm the ulculuion of the calibration curve HOWNU. Ihc told number of calibration standda that could be excluded m u t MI exceed20% of the toul number of tUmdud8 iqiccted. 13.5 Thc coml.tiOn ursfficicnt (R) for calibration curves generated must bc 20.992 (R' 20,985). If calibration mulu fall outside thsrc limits. Ihcn qpmp*td step# mum be taken to adjut inmUment operation. and Ihc rundvdior Ihcr8lcvmtrct of umpla ahould bc runalyzsd. 13.6 Retention tima b e e n rtmdrrdc md tunplea must not drift more than f 4 %within an d y l i c a l run. If rchtion time drift exceeds this limil within an vulytiul run ulcnthe LCI muat be nwlyzed. 14.0 C.leuluioar 14.1 UM tho followingequalionto calcularc the amount of PFOA found (in ngimL. bwd on polk uu)winp the studvd curve (linear regression parunetcrs) puwdby thc M u s Lynx aoRwuc pmgrun: - PFOA fwnd (nghnL) dope x DF x aliquot factor -DF frcor by whichthe hdvolumeWY diluted, if ncceuuy. Aliquot f m r - IO Exygen Research Page 5 7 of 65 .. ._ Page 123 of 140 ~ ~ ~~ Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 %em-h M e b d Numbor VooO1185 1 Memod of Me for the mu Doledn m atio c aMETHOD n of P a f l u o m u w i c Acid (PFOA) in Small I Munmd Liver by LCNSNS 14.2 POI rsmplw fOnj6d wilh h o r n m u n U of PFOA prior Io extraction. use tbo following equation to cdcullte the percent rccovay. 14.3 Use tb fol- equrlionlo m v a l the mount of PFOA found in nghL 10 - WSW). PFOA found (ppb) ample weight (g) Exygen Research Page 58 of 65 ~~ - Page 124of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 ANALYTICAL METHOD Mehod Number: VooOl786 Analytical Testing Facility: Approved By: Exygcn Rsreueh 3058 R-h Drive State College, PA 16801 Exygen Research TdPlpa: 7 Page 59 of 6S -_ Page 125 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 L m,u,nwx.MeTnoD Mcthod ofh d y a i a for tbc DUamidonofP~uoroostMotcAcid (PFOA) in Small MuMlrlScnun by LCIMSNS I .o s w p c Thia mahod ia to k anploycd for the ilolation and quvltitrtion of periluomwtanoic acid by High Pcrfn- Liquid Chronulogmphy couplal lo a landern Mass SpcchumetrisDdcslor(Lc/MS/MS)in d l mprrrrmlrerum 2.0 safely 2.1 Alwayl obrave d e pnctica. 2.2 COMlll the Opmprirtc MSDS before h l n d l i any chemical for proper safely plW4UboM. 3.0 SampleRequirmal 3.1 At lout I mL oftal umplc fmC X m S t i w . 3.2 No umple pmceaaing ia noodrd for MM I'I I umpla. However. frozen serum umplcr must to .Uowed D wmplctely thaw to room tsmpcrahm before use 3.3 Sample collection pmccdvsr will bc rpccifial in the sampling plan for thlr pmjm. 4.0 RupmtrradSlMddS - 4.1 W.ta HPLC @ 4.2 McUuool-HPLC@C /- 4.3 4.4 4.5 A~l~ilc-HPWgndc - Ammoni~mAcav~ A.C.S. R q c n l GI& - Psfluomoct.noic Acid Sigma-Aldrich 5.0 Irvtrumclltrad Equipment 5.1 A high pcrbnnmoe liquid chmrmtomh capable of pumping up 10 2 lolvcnta equipped with a vuiable volume injcetor capable of injecting 5-2Wl pL connectedIo a undcm M r u Spcchumctcr(LCIMSh4.S). 5.2 A dwiato wUect NW bu forpuk intcgdon and quanutotion. 5.3 Analylicalhlvrscapableof~dinpto0.OM)OIg. 5.4 50mL di8pUblcpolrpropylcncccntrifigc a h . 5.5 IS mL dispodle polypropylene CcDhifUgetubu. m e 5.6 Dirpoublcmicropipar(50-l00& 100-2OOuLI. 5.7 125-mL Ilpmw-mouth bottles. 5.8 2 mL clcu HPLC vial kil. 5.9 D*pornblepipetta. 5.10 Autopipctta (1OO.IM)o pL md 10-100 pL), with disposable tips. 5.11 Wrtm Sop Pak V u 6 cc (IdC I S SPE ur(ridgu. 5.12 SPB vlcuum mmifold. 5.13 Vonexa. Page 60 oj 65 Exygen Research Page 126 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 ' ' A1IALYllCAL YETHOD Method of Anrlysii for the Daorminuion of Perflwmoclanoic Acid (PFOA) in Small MMmd Sorum by LClMSlMS 5.14 Wriot-aaionhka. 5.15 Cmbihgc up.ble of lpinning I5 mL polypropylenetuba at 3000 rpm. 6.0 ChmnutolplphicSptm 6.1 Anrlyticrl Column: Plwphuc RP (Keystone Scientific), 2.I mm x SO mm. 5th - 2 (PM:82505-0S2l30) 6.2 Tanpsnturc: 30.C 6.3 Mobile Phus(A): 2 mMAmmoniumAcclate in Water 6.4 Mobile Phuc(e): Mnhrnol 6.5 GndicntRo&nm: Flow Ralc Yaw 0.0 35 0.3 1 .o 65 35 0.3 8.0 25 75 0.3 20.0 25 75 0.3 22.5 65 35 0.3 - 6.6 InjectionVolume: I5 pL (canbe incrrucd to as much as 50 #L) 6.7 Quantilation: Puk Area - extenul nandud calibration curve. 6.8 Run Time: 23 minutea. The 8bove conditionauc intendedu a guide and may be changed in order to optimizc the HPLC aptem. 7.0 MS/MSSystan 7.1 Modc: Elecmspny Neg.Cive MRM mode. monitoring41 3 + 369 m/z foi PFOA. The h v c conditiolu M intended u a guide and may be changed in order to optimize the MSMS :yuan. 8.0 P r e p d o n of Solutions 8.1 MobilePhuc 8.1.1 2 mM Mllllonium ~ c c u t ein water is prepared by adding 0.154 g of ammoniumWUICto loo0 mL of water. Altanatcvolumu m y be prspusd Exygen Research ... -_ Page 61 of 6s Page 127 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 E x y p llmuxh MahadNumbcrV0001786 I 1 ANALYTICAL .W:THOD Method of AnaIyci: for the DctmnhationofPerfluomwtnnoic Acid PFOA) in Small Mammal S m m by LCIMSIMS 9.0 StandadPrcpmtion 9.1 Standud StoddFortifiutionSolution 9.1.1 Rtpm a rtock solution of -100 pUmL of PFOA by weighing IO mg of analytical1Undud(comcted for purity) md dilute to I 0 0 mL with methum1 in 8 125-mL LDPE bottle. 9.1.2 A 1.0 p g h L fortification solution of PFOA ir prepared by bringing I mL of tho 100 pglmL solution to a GMI volume of I 0 0 wilh methanol in a 12) mL LDPE bottle. 9.1.3 A 0.1 p&nL fonificalion solution of PFOA ir prepared by bringing I O mL of tho 1.0 r@L solution to a find volume of LOO with mctbnol in a 125 mL LDPE bottle. 9.1.4 Thsstock and fortificationsolution$are to be stored in a rehigemor at approximrtcly 4*C ud arc itable for a maximum period of 6 months from chs date of prepantion. 9.2 StandardCalibrationS O ~ U ~ ~ O M 9.2.1 LUhlS/MS EllibrationNndpdr arc p r c p d m methanol via dilution of tho 0.1 pUmL fortificationwlution. 9.2.2 Ths following is a typical example: additional concentrations may be prepared u needed. Cauxnmtion Final ofFonification Volume Diluted to Concenlration Solution (ng/mL) (mL) (mL) (ndmL) I00 5.0 100 5.0 I00 2.0 IW 20 100 1 .o 100 I .o 5 .O IO 100 0.5 2.0 10 IW 0.2 1 .o IO 100 0.I 9.2.3 Sima dl Ulibdon rtnndudr in 125-mL LDPE narrow-mouth bottles nt 2'C to 6.C. up to rix months. 9.2.4 Altmuto volvmu d C O I I C W I ~ ~ O I Mof standards may be prepared as needed. 10.0 BuchSaUp 10.1 Each buob of umplcl exmusd (typiully 20 or less) must include at least one u n d calm1 and two untreated contmlr fonifkd at known concmtdonr (lab contml spike.)to verify procedural recovery for the batch 10.2 Requimncntlfor field md labontoryduplicntn Md spikes WIIIbe specified in the quality amuance plan for this project. Exygen Research Page 62 of 65 . Page 128 of 140 _______~ ~ Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen ProtocolNumber: PO000760 1 1 AMLYTICAL ,METHOD Mahod of And+ for ch DnamirrPtionof PcrflwrooclsnoicAcid (PFOAI in Small 11.0 SampleExlnction 11.1 11.2 11.3 11.4 I I .S 11.6 11.7 11.8 11.9 11.10 11.1 I Mururc 1 mL of sample inlo a SO mL polypmpyla~centrifugetubes (fonify u needed q l a c c lid and mix well). Note that d t m t e volumes of serum may be mouulod dopadingon the sample sizc available for use. Add wuu to the ramplc for a fuul volume of 20 mL. Cap lightly Voncx for -I minute. Tramfor 1 mL of the umple wing a disposable pipcnc into a IS mL dirpwblc CPIlrifugctube. Add S mL of rcczOnitrilculd W e for -20 minutes on a wrist-actionshaker Centrifugethe tvber U -3000 rpm for -5 minutcs. Deunt the nrpomntsnt inlo a SO mL disposable cmtrifugc tube and add 35 mL of w u a . Condition th CMSPE sadridga (1 g, 6 mL) by passing IO mL methanol followedby 5 mLof HPLC water (- 2 drop/ca). Do MI let column run dry - Lodthe umple on wnditionedCII SPE cadridge. Discard eluate. Elute with -2 mL of mcthulol. Collcct 2 mL of cluatc into a graduated 15 mL polypmpylonc mkifuge ~ b (fcinal volume 2 mL). Mpumpla usingclfftmspray LC/MS/MS. 12.0 Chromuopphy 12.1 lnjcct thc same LIIyIuo( of c u b ~Undudr, q l e and fortified sample into the LC/MS/MS sptcm. A calibntion It.ndard must precede and follow a11 d j z e d umpla. 12.2 Stpndrrdr of PPOA wmrponding lo 01 l u s t five or more concentration levels must be included in M uulyticd IC[. 12.3 An onlire sl of calibrationUUIdudI must be included Ithe begiMinp and at the end of a m p l e set Stvlduda must be interspersed betwan every S-ICl umpler. AI an d;omUivc. an cntirc 101 of calibration standards may be i n j d U th beginoing of st followed by calibration standards intmpeMdovary 5-10 r m p l a (lo account for a caond $elof standads). In citbcr cue. calibration rllndvdr must bc the fmt and last injection in a ample I d . 12.4 UIC limU dmdard c w a for quantitnlion. Linear standard curves are gcnaatedfor the d y t c by lmar r c p u i o n using Ilx weighting of peak area vmua calibration klmulud w m l r a t i o n Usingh l y n x 3.3 (or cquivalcntl IoftwMIystWl. 12.5 Sample rapcmc dwuld not sxcecd rundud ruponrcr. Any samples that a d dmdard rrrpolucr d w d d be hutbcrdilutcd md nurdped. Exygen Research Page 63 of65 Page 129 of 140 _ _ _ _ _ _ _ _ _ ~ ~~ ~ ~ ~~~ ~ ~~ ~_______~ Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number:PO000760 Exygm Rucuch MnhodNvobcr V w O 1 7 8 b 1 I ANALYTICAL MITHOD Method of Analylis for the Delamination of Perfluomoetanoic Acid (PFOA) in Small Mammal Scnun by LC/MS/MS 13.0 AcccpunScCritaia 13.1 c h r o m s t o ~must ahow a peak of a dauater ion at 369 m u hom a parent of 413 mu. Ihc 413 m u pumt corrcrpandato the PFOA anion, while the daughterion (369MU) r r p s e n utha loa of carbon dioxide. 13.2 Melbod b l h mwt not contain PFOA at levelr greater than the Log. If a blank con& PFOA It lovcli greater than IO ng/mL, then a new blank sample mwt beobtain& and tho e n h sa must be re-cxtloEted. 13.3 Rkovcria of control rpika and matrix rpiku mwt be between 70.130% of their kmwn valuea. If i control rpike falh outside the acceptable limits. the e n h 8cl of umplel ahould be rr-cxmEtod. Any mauix spike outside 70130% h u l d bo ovduited by the 8nItylt 10 dctnminc i f re-exIraction IS Wprrmtcd. 13.4 Any calibration sUdud found to be a atltiitical outlier by usins the Huge Ermr Tat, m y be exsludcd from thc CPlmLntion of the calibration CUNC. However, tho tow number of calibntion Wud~th.1 could be cxcludcd mud not cxcced 20% of the toul number of rt~dardi.n~jected. 13.5 Tba cornlalion Mefficient (R) for calibration c w a generated must be 20.992 (R' 20.98s). If calibration rerultr fall outside there limits. then ~ppropr*(s#@a must be taken to ldjusl inslrwncnt operation. and the 8mdard1or tho relevant set of m p l e r should be reanalyzed. 13.6 Mention time1 between itandub and samples must not drift more than t 4 K witbin an uulytical run. Ifrelention time drift exceeds this limit within M d y t i u l run then the rst must be runrlyzed. /- 14.0 Cdculaliwr 14.1 Use tha followingcqudon w alcuhte cho mount of PFOA found (in n@mL. bvcd on porL ma) w i q thc 11.adsrd c w e (linear mgrcsaion parameten1 generated by tbc h a Lynx MPWM program: PFOAfovod(nghnL)-&&~ . ' 'L x DF x aliquot facta - - slope DF &tor by which the find volume WM dilutcd. if nccersary. Aliquot fwtor 20 14.2 For umplcl foltified with known mounts of PFOA prior to extraction. YSK R c tho eov -following ay ('A) equalion to calculate the percent r a e o v n y . [toul d y t e found (nghnL) - d y t e foundin control (ng/mL)1 I oo d y t e ddcd(n&L) Page 6 of 7 Page 64 of 6s Exygen Research Page 130 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 hylm llrHvrh I Method of AMI+ ANALYTICAL %TIOD 1 uemodNumbcr vooo1786 for theDctcrminationof PortluomoctanoicAcid (PFOA) in Small M m l l Sawn by LCIMSIMS 14.3 UICIIIC followingqvuionto wnvm h e amount of PFOA found in ng/mL Io PPb. PFOA fouud @pb) - v m sample volume (mL) .. - ~ Exygen Research Page 65 of6S Page 131 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 3058 Research O& phone: 814-272-1039 State College, PA 16801 Fax: 814-231-1580 PROTOCOL WENDMENT Amendment Nwnber.L Effective Date: 01/19/05 Exygen Study Number; woo0760 C l h t Study Number. Page 1 of1 None P E X RIPTIONOF ED SECTION 1) AnaryUcal Procedure Summary V0001780:sedlon 9.1 2) Verification of Analytical Procedure A4Ewm2 1) Addta~~9.l:Section9.1.6,AltemabweightsofBtandardsmaybewedto prepare alternate concentrations of stock solutions as mcessary. Alternate levels of fortiRcationsolutions may also be prepared. 2) Low and highspiking levelsof the analytes for each matrix may be altered depending on sample stze available far exbaction andlor to cover analyte concentrations expected in the samples. RATIONAL5 1) Higherconcentralions of standardsneedto be preparedin order to spike the sample bottles at higher levels. 2) The samplesize available for smell mammalliver and semwas smaller than expected. Spiking at the prsdebrmlnedlevels h the protocol putsthesplked ancantration lawer than the detecbion limit Also, the analyte levels in the gnxlnd water samples are expecbd to greatly exceed the pre-determinedspiklng levels listed Inthe pmtocol. When the levels in the samples greatly exceed the spiking levels, an accurate recoveryvalue cannot be calculatedfor the QC sample. Higher spiking levels in the bottleswill c o w the anaJyte concentrationsexpeded Inthe water samples. IMPACT ON STUDY The LOQ is 100 ng/gfor a 0.1 g sampleof m Umammal liver and is 1000 ng/mLfor a 0.01 ml Sampb of small mammal serum. Higher levels of spiking for the water sampleswill ensure that more QC recoverydata can be used. LIBRARY IDWOO012268 Exygen Research Exygen QAU Review kl,I & ADMINISTRATIVE FORM Page 132 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 1 KsEA R c H . :-. 3058 Research Drive Phone: 814-272-1039 State College, PA 16801 Fax: 814-231-1580 Amendment Number: Effective Date: Exygen Study Number PROTOCOLAMENDMENT 2 03/07/05 PO000760 Client Study Number: Page 1 of 1 None DESCRIPTION OF AMENDED SECTION Report, page 11 of 65 fYrmwm Instead of one final report, interim reports will be issued. RATlONALE Due to the excessive sizes of the data sets, interim reports will be issued to allow the dient to receive data in a timelier uwmr. c$+m IMPACT 0N STUDY The dient will be able to receive and reviewthe data more quickly. LIBRARY IO: VOOO1226-8 Exygen Research ADMINISTRATIVE FORM Page 133 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 -0 W ueruRvm:wmrpge RECEIMI)fIrE JUi. 2. 9:m PRINTTIM nDMlNlglRAm~ JUi. 2. 9:3EWl Exygen Research Page 134 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 JUL.ZS.ZBBS 8 : M MYGEN RE- No.774 P.4 Amendment Number: Effecthrs Date; ExygenStudy Number 3058 Research Orive Phone: 814-272-1039 State CoUege, PA16801 Fa: 814,231-1580 H PROTOCOL AMENDMENT - 07/18/05 P780 Client Study Number; Pag.1 dl NA - PESCRIPTION OF AMENDED SECTION Verificationof Analytfcsi Procedure,page t 0 ofprotocd. PaJlENOEO TO The field dupiicate can be used for the laboratoryspikes and replicate Wen the primary sampie volume Is limited. ii -. -. pATlUNAL me smpb size for a water sampk mL sample site requires raextmmn any reason, there would not be enough of the primary sample to w e a t two labamtar spikesand a rwliccate. The field duplicate is technically the same sample as the prlmar sample and iherefors,can be used for taboratory spikes and replicates as nersdrsd. JVlPAcr ON STUDY No mzgatiie impact on the study, Using the dupflcatesample allows for the ful QC c thm sample site to be completed. / Exygen Research Page 135 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 RESEARCH 3058 Research Drive Phone: 814-272-1039 State College, PA 16801 Fax: 814-231-1580 Amendment Number: Effective Date: Exygen Study Number PROTOCOLAMENDMENT 5 09113/05 P760 Client Study Number: Page 1 of 1 NA DESCRIPTION OF AMENDED SECTION Analytical Procedure Summary: V0001780:"Method of Analysis forthe Determination o Perfluoroooctanoic Add (PFOA) inWater by LCIMSIMS." AMENDED TO The reanalysis of two water samples, DFOS-GW-I31R-0-040126 and DFB-GW-135L-0. 040121 will be performed using the following procedure: An aliquot of the water sample is spiked with the surrogate standard andlor other matri) spike compounds as appropriate. Add methanol to the fortified aqueous sample. Mi) and aliquot for LClMSlMS analysis. The LC conditions include the use of a Betasil C18 2.1 x 50 mm column, a gradient of 30% B to 100% B in 10 minutes, where A=2mN ammonium acetate in water and B=Methanol (6 rnin equilibration). Use a 5 pL injectior volume and a 0.3 mUmin flow rate. LC conditions may be further modified a6 appropriate for the system being used. An interim report will be issued that will include the exact details of the reanalysis. RATIONALE DFOS-GW-I31R-0-040126 and DF8b-GW-135L-0-040121 could not be reportec quantitatively in interim report #3 due to quality control sample failures. The study director and sponsor have requested reanalysis using the above conditions. IMPACT ON STUDY No negative impact on the study. More usable data may be obtained. Study Director Signatye llrincipal Investigator %nature hllA Exygen ManagementSinatura Sponsor Signature (if required) Date . Date / Date Dale Exygen QAU Review m.+ qh3/Or LIBRARY ID: VOoO1226-8 -~__ ADMINISTRATIVE FORM Exygen Research Page 136 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 - PROTOCOLAMENDMENT e RECEIVED T I M SEP.14. 16:43AM Exygen Research PRINT TIME SEP.14. iB:44AM Page 137 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 TO: *88142311588 P: 2'2 twin No.294 P.2 3058 Raearch Drive Phone: 814-272-1039 State CdleEe, PA 16801 Fax: 814-231-1580 RESEARCH Amendment Number: Effective Date: Exygen Study Number PROTOCOL AMENDMENT 5 OW1 3/05 P760 Client Sbdy Number: Pagelofl NA DESCRIPTIONOF AMEN= SECTION AnaMcal Procedure Summanr: VOOO178O:%iefhod of Analv'sls fa the Determinationof Perfl;rCmwdanoicAcid (PFOA) inWater by LCIMSMS.' __ AMENDED TO The reanalysisof two water samples, D F O W - I 31RoO40128 and DFB-OW-135L-UOW121 wit be performed using the fdlowing procedure: An aliquot of the water sample is spiked with ?hesurrogatestandard andlar othw matrix spike compounds as appropriate. Add methanol to the f d e d aqueous sample. Mi and aliquot for Lc(MSIMSanalpiis.. The LC conditionsindudethe use ofa Betasif CW, 21 x 50 mm column. a gradient bf 30% B to 100% B in 10 minutes, where A=2mM ammonium acatate in water and kMethanol(6 min equilibration). Use a 5 pL injection - - I = I RECEIVED TIME SEP.14. 18:3?w1 --_. Exygen Research PRINT TIME SEP.14. 16:38Fy1 Page 138 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 EfKemR E S E A R C H 3058 Research Drive Phone: 814-272-1039 State College, PA 16801 Fax: 814-231-1580 0 PROTOCOLAMENDMENT Amendment Number: 6 Page 1 of 1 Effective Date: 09120105 Exygen Study Number P760 Client Study Number: NA DESCRIPTION OF AMENDED SECTION The identification of the two water samples listed in protocol amendment No 5 arc incorrect. AMENDED TO The two water samples for reanalysis should be DF09-GW-131R-0-040120 anc DFBb-GW-135R-O-O4012 1. RATIONALE The correct samples are identified for reanalysis. 0 IMPACT ON STUDY No negative impact on the study. Study Director Signalyra , , %ipal JA Investigator Sighture Exyen M a g w n a t u r e SponsorSignature(if required) 1) LIBRARY ID: VOOO1226-8 Exygen Research Dale Date / Date U-JtP-4,' Date Date ExygenQAU Review h . H 9/zo/Or ADMINISTRATIVE FORM Page 139 of 140 Interim Report #24 - Analysis of Water, Sludge, and Sediment Exygen Study No.: PO000760 CHEM EHSR 236 1B 651 733 1958 " 5Ep.a.ZRt5 2 : m 09/20 '05 13:-4-6--N- 0.920 02/02 m.380 P.2 0 .L c AmendmentNmbw EffectIw Dale: Exyoen Study Number PRotOCa AMENDMENT 6 o9/20/05 P760 Qlent Study Number: hgeIof1 NA LIBRARY1 O : V w Q l ~ RECEIVED TIME SEP.28. 3:BlPM Exygen Research ADMyJISTRA~~M PRINT TIME .YP.28. 3:02PM Page 140 of 140