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INTERIM REPORT #26 - Analvsis of Groundwater 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 REOUIREMENTS 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 January 9,2007 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 StudyNumber: PO000760 Total Pages: 119 Interim Report #26 - Analysis of GroundwaterSamples 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. Exygen Research Study Director Weston Solutions, Inc. Michael A. Santof Sponsor Repres tative 3M Company Exygen Research Date Page 2 of 119 Interim Report #26 - Analysis of Groundwater Samples 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 Date Inspected Date Reported to Date Reported to Principal Exygen Date Reported to Investigator Management Study Director 41. RawData& Interim Analflcal Report 12/28,29/06 01/09/07 0 1/09/07 01/09/07 1Note: 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 119 Interim Report #26 - Analysis of Groundwater Samples 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: 6J Chas Simons Operations Mana Exygen Research Exygen Research Facility Management: / Richard A. Date h Study Dire or, Weston Solutions,Inc. t- JaisimhaTeyJari P.E., DEE Weston Solutions, Inc. Sponsor Representative, 3M Company: / Director of Regulatory Affairs Exygen Research Date I Page 4 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen StudyNo.: PO000760 STUDY IDENTIFICATION 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 PROTOCOL NUMBER: PO000760 EXYGEN STUDY NUMBER: PO000760 TYPE OF STUDY: Residue SAMPLE MATRIX: Ground Water 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: 11/24/06 Interim Analytical Termination Date: 12/06/06 Interim Report Completion Date: 01/09/07 Exygen Research Page 5 of 119 ~ Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.:PO000760 The Study Director for this project is Jaisimha Kesari at Weston Solutions, Inc. The following personnel fiom Exygen Research were associated with various phases of this interim portion of the study: Name Chas Simons John Flaherty Karen Risha Amy Sheehan Mark Ammerman Mindy Cressley Ellen Dashem Chrissy Edwards Krista Gallant Title Operations Manager Vice President Laboratory Supervisor Associate Scientist Sample Custodian Technician Technician Technician Technician Exygen Research Page 6 of 119 Interim Report #26.Analysis of GroundwaterSamples Exygen StudyNo .. PO000760 TABLE OF CONTENTS 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 ............................................................................................................... 11 3.0 INTRODUCTION....................................................................................................... 11 4.0 ANALYTICAL TEST SAMPLES.............................................................................. 11 5.0 REFERENCE MATERIAL........................................................................................ 12 6.0 DESCRIPTION OF ANALYTICAL METHOD ........................................................ 13 6.1. Extraction Procedure ............................................................................................... 13 6.2 Preparation of Standardsand FortificationSolutions............................................... 13 6.3 6.4 ICnhsrtorummaetongt rSaepnhsy.it.i..v....i.t.y............................................................................................................................................................................................... 14 14 6.5 Descriptionof LCMSMS Instrument and Operating Conditions........................... 15 6.6 Quantitationand Example Calculation..................................................................... 15 7.0 EXPERIMENTAL DESIGN ...................................................................................... 16 8.0 RESULTS ................................................................................................................... 17 9.0 CONCLUSIONS......................................................................................................... 17 10.0 RETENTION OF DATA AND SAMPLES ............................................................. 17 Exygen Research Page 7 of 119 Interim Report #26 . Analysis of Groundwater Samples Exygen StudyNo .. PO000760 Table I. Table II. Table ID. LIST OF TABLES Pag;e Summary of PFOA in Groundwater Samples................................................. 19 Matrix Spike Recovery of PFOA in Groundwater Samples........................... 20 Surrogate Spike Recovery of 13CPFOA in Groundwater Samples................21 Exygen Research Page 8 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Figure 1. Figure 2. Figure 3. Figure 4. Figure 5. Figure 6. Figure 7. Figure 8. LIST OF FIGURES Pane Typical Extracted Calibration Curve for PFOA in Reagent Water ................23 Extracted Standards of PFOA in Reagent Water, 0 ng/L and 25 ng/L, Respectively................................................................................................... .24 PFOA in Reagent Water, 50 ng/L Fortified Reagent Spk A, and 500 ng/L Fortified Reagent Spk B, Respectively ........................................... 25 Chromatogram Representing a Groundwater Sample Analyzed for PFOA (Exygen ID: C0222023, Data Set: 112406C)...................................... 26 Typical Extracted Calibration Curve for 13CPFOA in Reagent Water ..........27 Extracted Standardsof 13CPFOA in Reagent Water, 0 ng/L and 25 ngL, Respectively................................................................................................... .28 13CPFOA in Reagent Water, 50 ng/L Fortified Reagent Spk A, and 500 ng/L Fortified Reagent Spk B, Respectively ........................................... 29 Chromatogram Representing a Groundwater Sample Analyzed for I3CPFOA (Exygen ID: C0222023, Data Set: 112406C)................................ 30 Exygen Research Page 9 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.:PO000760 LIST OF APPENDICES Appendix A Study Protocol PO000760 (Exygen Study No. P0000760) with Analytical Method V0001780: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS", Protocol Amendments, and Deviations ..........................31 Exygen Research Page 10 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 1.0 SUMMARY Exygen Research extracted and analyzed groundwater samples for the determination of perfluorooctanoic acid (PFOA) according to Exygen Method VOOOl780 (Appendix A), The limit of quantitation (LOQ) for PFOA in ground water was 25 ng/L. Analytical results for the analysis of PFOA in groundwater samples are summarized in Table I. Fortification recoveries for PFOA in the groundwater samples are detailed in Table 11. The overall average percent recovery k standard deviation for PFOA in groundwater samples was 96 f 22%. Fortificationrecoveries for the 13CPFOA surrogate spike in the groundwater samples are detailed in Table 111. The average percent recovery k standard deviation for the 13CPFOA surrogate spike in the groundwater samples was 99 f 14%. In addition to the groundwater samples, a quality control reagent blank, reagent spike A, and reagent spike B were included in each set. 2.0 OBJECTIVE The objective of the analytical part of this study was to determine levels of perfluorooctanoic acid (PFOA) in groundwater according to Protocol PO000760 (Appendix A). 3.0 INTRODUCTION This report details the results of the analysis for the determination of PFOA in groundwater using the analytical methods entitled, "V0001780: Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Water by LCMSMS." The study was initiated on November 05, 2004,when the study director signed protocol number POOOl131. The analytical start date for this interim report was November 24, 2006,and the analybcaltermination date for this interim report was December 6,2006. 4.0 ANALYTICAL TEST SAMPLES Eleven groundwater samples (Exygen ID C0222023 - C0222033) were received on wet ice on November 21,2006 fiom Tim Frinak at Weston Solutions, Inc. Eight groundwater samples represented two groundwater sampling sites and associated field quality control Exygen Research Page 11 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 samples. Three water samples represented one trip blank and two trip blank spikes. All samples were logged in by Exygen personnel and placed in refiigerated 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 (SP0008065), was purchased from Oakwood Products, Inc. and was received at Exygen on September 8, 2006. The surrogate spiking standard, 13C labeled perfluorooctanoic acid (13C PFOA SP0007767), was received at Exygen on June 13,2006 fiom the 3M Company. The available information for the reference materials is listed below. PFOA was stored ambient and 13CPFOA was stored fkozen. Compound PFOA 13C PFOA Exvnen Inventory No. SP0008065 SP0007767 Lot # Y16G 3507-277 Purity C%) 98 97.6 Expiration Date 09/08/08 07/15/10 The molecular structure of PFOA is given below: PFOA Chemical Name: Perfluorooctanoic acid Molecular Weight: 414 Transitions Monitored: 413 +369 (for quantification)and 4 13 +2 19 (for confirmation) Structure: F F F F,/\/\/\ OH F F F 13C PFOA Chemical Name: 1,2- 13C perfluorooctanoic acid Molecular Weight: 416 Transition Monitored: 415 + 370 Structure: Exygen Research Page 12 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 6.0 DESCRIPTION OF ANALYTICAL METHOD The analytical method "V0001780: Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS" was used for the groundwater samples in this study. 6.1. Extraction Procedure 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 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 centrihge tube. Each sample was analyzed by LC/MS/MS electrospray. 6.2 Preparation of Standards and Fortification Solutions A stock standard solution of PFOA was prepared as specified in Exygen method VOOOl780. The stock standard solution was prepared at a concentrationof 10,000 p g / d by dissolving 1.0 g of the standard (corrected for purity and salt content, if necessary) in acetonitrile. From this solution, a 1000 pg/mL fortification standard solution was prepared by taking 10 mL of the stock and bringing the volume up to 100 mL with acetonitrile. By taking 10 mL of the 1000 p g / d fortification standard and bringing the volume up to 100 mL with acetonitrile, a 100 pg/mL fortification standard was prepared. By taking 10 mL of the 100 pg/mL fortification standard and bringing the volume up to 100 mL with acetonitrile, a 10 pg/mL fortification standard was prepared. By taking 10 mL of the 10 pg/mL fortification standard and bringing the volume up to 100 mL with acetonitrile, a 1.0 pg/mL fortificationstandard was prepared. By taking 10mL of the 1.0 pg/mL fortification standard and bringing the volume up to 100 mL with acetonitrile, a 0.1 pg/mL fortification standard was prepared. By taking 10 mL of the 0.1 pg/mL fortification standard and bringing the volume up to 100 mL with acetonitrile, a 0.01 pg/mL fortification standard was prepared. Exygen Research Page 13 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.:PO000760 A stock standard solution of 13CPFOA was also prepared as specified in the raw data. The stock standard solution was prepared at a concentration of 100 pg/mL by dissolving 0.01 g of the standard (corrected for purity and salt content, if necessary) in methanol. From this solution, a 10 pg/mL fortification standard solution was prepared by taking 10 mL of the stock and bringing the volume up to 100 mL with methanol. By taking 10 mL of the 10 pg/mL fortification standard and bringing the volume up to 100 mL with methanol, a 1.0 pg/mL fortification standard was prepared. By taking 10 mL of the 1.0 pg/mL fortification standard and bringing the volume up to 100 mL with methanol, a 0.1 pg/mL fortification standard was prepared. By taking 10 mL of the 0.1 pg/mL fortification standard and bringing the volume up to 100 mL with methanol, a 0.01 pg/mL fortification standard was prepared. A set of standards containingPFOA and 13CPFOA were prepared in water and processed through the extraction procedure, identical to samples. The following concentrations were prepared: Conc. of Fort Fort Solution Volume (ng/mL)' (pL) 0 0 10 100 10 200 10 400 100 100 100 200 1 00 400 of PFOA 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 refigerator (4" k 2OC) when not in use. Documentation of standard preparation is located in the raw data package associated with this interim report. 6.3 Chromatography Quantification of PFOA was accomplished by LC/MS/MS electrospray. The retention time of PFOA was -10.5 min. Peaks above the LOQ were not detected in any of the reagent blank samples corresponding to the analyte retention time. 6.4 Instrument Sensitivity The smallest standard amount injected during the chromatographic run had a concentrationof 25 ng/L of PFOA. Exygen Research Page 14 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen StudyNo.:PO000760 6.5 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: Therm0 Fluophase 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: Analvte PFOA PFOA Confirm Ion 13C PFOA Mode negative negative negative Transition 4M13on-+ito3re6d9 413 +219 415 + 370 Approximate Retention Time 0 -10.5 min. -10.5 min. -10.5 min. 6.6 Quantitation and Example Calculation Fifteen microliters of sample or calibration standard were injected into the L C M S M S . 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 fiom the equations below. Equation 1 calculated the amount of analyte found (in ng/L, based on peak area) using the standard curve (linear regression parameters) generated by the Analyst software program. Exygen Research Page 15 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Eauation 1: Analyte found (ng/L) = (Peak area - intercmt) 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 13C PFOA prior to extraction, Equation 2 calculated the percent recovery. Eauation 2: Recovery (%) = lanalvte found (ng/L) - analvte in control (nrz/L)) x 100% amount added (ng/L) An example of a calculation using an actual sample follows: Water sample Exygen ID C0222023 Spk C (Set: 112406C), fortified at 250 ng/L with PFOA where: peak area - intercept - slope - dilution factor - ng/L PFOA added (fort level) - amt (n&) in corresponding sample = 53 1829 6960 822 1 250 358 (Set: 112406C) From equation 1: Analyte found (ng/L) = 1531829 - 69601 x 1 822 From equation 2: % Recovery = 639ng/L = (639 - 358ngL) x 100% 250 = 112% NOTE: Numbers may differ slightly fiom raw data due to rounding. 7.0 EXPERIMENTAL DESIGN 13C PFOA was used as a surrogate for all the samples. 13C PFOA was added to the sample collection bottles in the laboratory before being shipped to the field for sampling. For 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 samples were filled to a 200 mL volumetric fill line in the field. Exygen Research Page 16 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 The samples were extracted in one set. The set included one reagent blank, two reagent blanks fortified at known concentrations. The set also contained two sample sites and the field blank and the field blank spikes collected for the groundwater samples. For each site, a sample, a field duplicate and two-matrix field spikes were collected. For each site, a laboratory duplicate and two laboratory matrix spikes were also extracted. 8.0 RESULTS Analytical results for the analysis of PFOA in the groundwater samples are summarized in Table I. Accuracies were assessed for each sample by reviewing the individual quality control results obtained for each sample site. In most cases, there were two laboratory and two field spike recovery results available for each sample site that were used to assess the accuracy. In instances of failed laboratory or field spikes, recoveries associated with other spikes were used to assess sample accuracy. Fortification recoveries for PFOA in the groundwater samples are detailed in Table 11. The average percent recovery standard deviation for PFOA in the groundwater samples was 96 f 22%. Fortification recoveries for the 13C PFOA surrogate spike in the groundwater samples are detailed in Table 111. The average percent recovery -f: standard deviation for the 13CPFOA surrogate spike in the groundwater samples was 99 k 14%. In addition to the ground water samples, a quality control reagent blank, a low reagent spike A, and a high reagent spike B were included in each set. 9.0 CONCLUSIONS The groundwater samples were successfully extracted and analyzed for PFOA according to analytical method VOOO1780. 10.0 RETENTION OF DATAAND 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 Standards40 CFR 792. Exygen Research Page 17 of 119 Interim Report #26 -Analysis of GroundwaterSamples Exygen Study No.:PO000760 TABLES Exygen Research Page 18 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: pooO0760 Table I. Summary of PFOA in Groundwater Samples Cbnt Cm22M7 C0222027 Rep cOa2M28 CO222031 I OALOW-RESW01-0-081117 DAL-GW-RESW01-0-061117. DALGWRESWOl-DM61117 DAL-GW-RESWMQOG1117 DAL-GW-RESW02-0-06111'r DAL-GW-RESW02-DBO61117 DAL-GW-TRIP-O-El 117 Analyte Found (ppt, nHL) C8 Acid PFOA A s r r r d Accuracy 2580 30 2430 30 m0 30 ND 30 Exygen Research Page 19 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Table 11. Matrix Spike Recovery of PFOA in Groundwater Samples P - Sample Descrlptlon DAL-GW-RESWOI-0-061117 (C0222023 Spk C, 250 n@L Lab Spike) DAL-GW-RESWO1-0-061117 (C0222023 Spk D, 5ooo ng/L Lab Splke) DAL-GW-RESWO1-LS-061117 (CO222025,0.25 ppb Field Splke) DAL-GW-RESWOl-HS-061117 (C0222026,5 ppb Field Spike) DAL-GW-RESW02-0-061117 (C0222027Spk E,250 ng/L Lab Spike) DAL-GW-RESWO2-0-061117 (C0222027 Spk F, 5000 n@LLab Splke) DAL-GW-RESW02-LS-061117 (CO222020,0.25 ppb Fldd Spike) DAL-GW-RESWO2-HS-061117 (C0222030,5 ppb Field Splke) DAL-GW-TRIP-LS-061117 (C0222032.025 ppb Field Spike) DAL-GW-TRIP-HS-061117 (CO222033,5ppb FieldSpike) ____ Amount Amt Found Spiked in Sample (na/L) (ngL) P 250 358 5000 358 250 358 5000 358 250 2580 5000 2580 250 2580 5000 2580 250 ND 5000 ND C8 Acid PFOA Amount Recovered 638 5310 667 3160 3060 7150 2630 6190 269 5260 - Recovery 112 99 124 56 . 91 72 1oa 105 Average: 96 Standard Deviation: 22 'Sample residue exceeds the spiking level significantly; therefore, an accurate recovery value cannot be calculated ND = Not detected at or above the Limit of Quantitation (LOQ) of 25 ngL. Note: Since this summary table shows rounded results, recovery values may vary sllghtly from the values In the raw data. Exygen Research Page 20 of 119 Interim Report #26 -Analysis of GroundwaterSamples Exygen Study No.: PO000760 Table 111. Surrogate Spike Recovery of 13CPFOA in Groundwater Samples - - Exysen - ID Sample Description C0222023 Spk C DAL-GW-RESWO14 - 0 6 1117 C0222023 Spk D DAL-GW-RESWO1-0-061117 C0222023 DAL-GW-RESWO1-0-061117 C0222023 Rep DAL-GW-RESWOl-0-061117 C0222024 DAL-GW-RESWO1-DBO61117 C0222025 DAL-GW-RESWO1-LS-061117 C0222026 DAL-GW-RESWO1-HS-061117 C0222027 Spk E C0222027SpkF C0222027 C0222027 Rep C0222028 C0222029 C0222030 DAL-GW-RESWO2-0-061117 DAL-GW-RESW02-0-061117 DAL-GW-RESW02-0-061117 DAL-GW-RESWO2-0-061I 1 7 DAL-GW-RESW02-DE061117 DAL-GW-RESW02-LS-061117 DAL-GW-RESW02-HS-061117 C0222031 C0222032 C0222033 DAL-GW-TRIP-0-061117 DAL-GW-TRIP-LS-061117 DAL-GW-TRIP-HS-061117 l s ~ ~ ~ ~ Amount Spiked (n@// Amount Recovered (nglL) Recovery (%) 750 811 108 5500 5860 107 500 517 103 500 517 103 500 475 95 250 308 123 5000 2900 58 750 756 101 5500 5580 101 500 485 97 500 458 92 500 504 101 250 244 98 5000 3930 79 500 524 105 250 263 105 5000 5280 106 Average: 99 Standard Deviation: 14 Note: Since this summary table shows rounded results, recoveryvalues may vary slightly from the values In the raw data. Exygen Research Page 21 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 FIGURES Exygen Research Page 22 of 119 Interim Report#26- Analysis of GroundwaterSamples Exygen Study No.: PO000760 Figure 1. Typical Extracted Calibration Curve for PFOA in Reagent Water 112406C P7W-1131 W h r R.r.rdb (PF0A):"Lind` RagiHion ("1 l i m i q h t i n q ) : y = 8 2 2 x+G.-36.+003 (1=0.9991) 8.3.6 8.0.5 7.5.6 7.0.5 G.5.6 G.O.6 5.5.6 5.0.6 u1 c c 4.5.6 a 0 4.0.6 g 4 3.54 3.0.6 2.5.6 2.0.5 1.5.6 1. O d 5.0.4 On 100 200 300 400 m mo 700 800 Conantration. n d L 900 laxl Exygen Research Page 23 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Figure 2. Extracted Standards of PFOA in Reagent Water, 0 ng/L and 25 ng/L, Respectively IX C I I ~ P~FO. A(Standrrd)413.0#69.0 rmu - sample I of29 from 11240dCR.~ff (peak notfound) 1500- (0 8 1m: - m c c 500 - 2.6l M 0' W 6.18 -7.94 8.06 r 0.w. -1 Exygen Research Page 24 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Figure 3. PFOA in Reagent Water, 50 ng/L Fortified Reagent Spk A, and 500 ng/L Fortified Reagent Spk B, Respectively I-Ut a E t r O # .ffm R * r k U O W t t ) 4 l 3 M b l O s ~-W~MP#C ) O f MfrOM ll2406CWiW Area: 400counb f i g h t : l.C4m+Q@2cpRrE lassin 3 M .I 10.46 0- 1 2 3 4 5 G 7 8 0 10 I1 12 13 14 15 16 17 I Tima. min Riagent Spk A - PFOA(OC)413.OW6~.0 m u - sample 10 of34from 112@3C.wiW Ana: 40888 oounts Haiqht: 3.11e+W3 ops RT: 10.4 min I O 44 I Time, min Raapenl Splc B . PFOA(OC)413.0WG9.0 a m u . rampla 11 of34from 112@3CmiW Araa: 42115~34counts H d q M 2.6Ga+W4 cpr RT: 10.4 min 10.46 Time, rnin Exygen Research Page 25 of 119 Interim Report #26 -Analysis of GroundwaterSamples Exygen Study No.: PO000760 Figure 4. Chromatogram Representing a Groundwater Sample Analyzed for PFOA (Exygen ID: C0222023, Data Set: 112406C) 2R.4 - 1.9.4 - 1.84- 1.7.4- 16.4- 16.4- 1.4.4 - 13.4- W 12r4- E 1.1.4- g 1.0.4- m -E Qooo.0- - Bmo.0 - 7000.0 - 0000.0 - 5 m . o 4ooo.o m . 0- mw.0 - 1000.0 0.0 J n Time. min Exygen Research Page 26 of 119 Interim Report #26 - Analysis of GroundwaterSamples Exygen Study No.: PO000760 Figure 5. Typical Extracted Calibration Curve for 13CPFOA in Reagent Water 112-C P700.1131 Water Rcsrdb (13C PFOA): 'Lineat' RiprelrionC l lx" miphtinp): y = 724x+ 0.00017(r = 0.9979) - 722.6 7.02.6 - O D d 4.56 - 4.02.6 - c a -- d 3.62.6 e 4 3.02.6 0 I00 700 Conanhation. ng/L 1wO Exygen Research Page 27 of 119 Interim Report #26 -Analysis of GroundwaterSamples Exygen Study No.: PO000760 Figure 6. Extracted Standards of 13CPFOA in Reagent Water, 0 ng/L and 25 ng/L, Respectively 8.08 fin- - 1 2 3 4 6 0 7 8 9 10 11 12 13 14 16 I O 17 Time, - I I XC112qw-1.13C PFOA(St~ndrrd)415.013700 a m u . rrmpli 2 of34from 112-C wiff A I ~ J 20179 counb Haight. 1.54.- cps RT. 1 0 4 min 10.45 Time. min Exygen Research Page 28 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Figure 7. 13C PFOA in Reagent Water, 50 ng/L Fortified Reagent Spk A, and 500 ng/L Fortified Reagent Spk B, Respectively 0.08 0 lpn. # 5.0 c -c c 0 a 1 2 3 4 5 6 7 8 0 10 11 12 13 14 15 16 17 Time, fln _I I Reagent Spk A. 13C PFOA(OC)415.0t370.0 amu. sample I O of 34frmm 112400C mff Area.42- nnunk Hdpht 32!2e+003 opr R T 10.4mln 10.46 1 Time. n!& ~ Reagent Spk B . 13C PF0A(0C)415.0/3?0.0 amu. sample I ~ o f 1 3 4 l r o m112406C wff Area: 373725 counk Height. 2BOe+W4 nps RT 10 6 min 10.46 Time, min Exygen Research Page 29 of 119 Interim Report #26 -Analysis of GroundwaterSamples Exygen Study No.: PO000760 Figure 8. Chromatogram Representing a Groundwater Sample Analyzed for 13CPFOA (Exygen ID: C0222023, Data Set: 112406C) Time. min Exygen Research Page 30 of 119 Interim Report #26 - Analysis of GroundwaterSamples ~~ Exygen Study No.: PO000760 APPENDIX A Study Protocol PO000760 (Exygen Study No. P0000760) with Analytical Method, Protocol Amendments, and Deviations Exygen Research Page 31 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.:PO000760 Exygen Protocol N u m k 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 perform in^ Laboratory Exygen Research 3058 Research Drive State College, PA 16801 Phone: (814) 272-1039 - L- .~ Swnsor ReDresentative: Michael A. Santom Director of Regulatory Affairs 3M Building 0236-01-B-10 St. Paul, M N 55144 Phone: (651) 733-6374 Page I 0165 -_ Y Exygen Research Page 32 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen StudyNo.:PO000760 Exygcn Protocol Number: PO000760 DISTRIBUTION: 1) Jaisimha Kesari, StudyDirector, Weston Solutions 2) John M.Flaherty, Principal Investigator, Exygen Research 3) Michael A. Santoro, Sponsor Representative, 3M Company 4) Exygen Research Quality Assurance Unit Exygen Research Page 33 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.:PO000760 Exygen Protocol Number: PO000760 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 ProtocolNumber:PoooO760 APPROVALS c Weston Solutions w dzia ' Michael A. Sayt6ro. SponsorRepresentative 3M Company ,Mm M. Flahcrty, principal Investigator / Exygen Research I' ., - ident, Facility Management #A? Date - Exygen Research Page 3 of 65 - - . Page 34 of 1 19 Interim Report #26 - Analysis of GroundwaterSamples ~ ~~ Exygen Study No.: PO000760 Exygen F'rotocol Number: PO000760 TABLE OF CONTENTS TITLE PAGE ...................................................................................................................... DISTRIBUTION ................................................................................................................. PROTOCOLAPPROVAL................................................................................................................................ 3 TABLE OF CONTENTS....................................................................... INTRODUCTION.................................................................................. TESTMATERIAL............................................................................................................................................ 5 SURROGATEFIELD SPIKE COMPOUND........................................................................................ OBJECTIVE..................................................................................................................................................... 6 TESTINGFACILITY....................................................................................................................................... 6 STUDY DIRECTOR ............................................................................................................... SPONSOR REPRESENTATIVE.................................................................. PRINCIPAL INVESTIGATOR... ......................................................................................................... 7 PROPOSED EXPERlMENTALSTART AND TERMINATIONDATES. IDENTIFICATIONAND JUSTIFICATIONOF THE TEST SYSTEM ......................................................... 7 SAMPLEPROCUREMENT,RECEIPT AND RETENTION......................................................................... 8 SAMPLE IDENTIFICATION ....................................................................................................... 8 ANALYTICAL. PROCEDURESUMMARY................................................................................................... 9 VERIFICATIONOF ANALYTICALPROCEDURE...................................................................................... 9 METHODFOR CONTROLOFBIAS................................................................................ STATISTICAL METHODS ............................................................................................................................. I 1 GLP STATEMENT .......................................................................................................................................... I 1 REPORT ........................................................................................................................................................... I I SAFETY AND HEALTH ........................................................................................................................ AMENDMENTSTOPROTOCOL............ ................................................................................................ 13 DATA RECORD KEEPING ...... ................................................................................................................ 13 QUALITY ASSURANCE .......... RETENTION OF DATA AND ARCHIVING ................................................................................................. I4 APPENDIX I, ANALYTICALMETHODS ...................... - _.___-- ___- Page 4 of65 Exygen Research Page 35 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Exygcn Protocol Number: PoooO760 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 Lc/MS/MS for the 3M Decatur Monitoring mwn. 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 pduorooctanoic acid (PFOA) and was purchased from Sigma-Aldrich. PFQA ChemicalName: Pduorooctanoic acid Molecular Weight: 414 Lot Number: 231 16HB PtUity: 97.64% TransitionsMonitored. 413 +369 (for quantification)and 413 +219 (for confirmation) structure: FFFF SURROGATE FiELB SPIKE COM.PQI.ND The surrogate field spiking compound is "C labeled pduorooctanoic acid (I3CPFOA) and was purchasedby 3M from Perkin-Elmer Life and Analytical Sciences. "C PFOA ChemicalName: 1,2-13Cperfluorooctanoicacid Molecular Weight: 416 Lot Number: 3507-195 Purity: 97% Transition Monitored: 415 -+ 370 Exygen Research Page 36 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 rc OBJECTIVE "he 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: V0001780: V0001781: V0001782: V0001783: V0001784: V0001785: 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 LCIMS/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 LCIMSIMS" "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Small Mammal Serum by LCIMSIMS" 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. 1400Weston Way West Chester, PA 19380 Phone: (610) 701-3761 Fax: (610) 701-7401 j.kesari@westonsolutions.com - Page 6of6S Exygen Research Page 37 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 . Exygcn Pmtocol Number: P0000760 P SPONSOR REPRESENTATIVE Michael A. Santoro 3M Company Directorof RegulatoryAffairs 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@cxygen.com PROPOSEDEXPERIMENTALSTARTAND "EWWNATION r DATES It is proposed that the analytical porIion 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 JUSTIFICATEONOF TNE TESTSYSTEM The followingare the test systems for this study: Water (groundwaterand surfacewater) Soil sediment Fish Clams Vegetation SmallMammalLiver SmallMammalSerum The samples will be collectedby Weston Solutions. The control samples will be purchased and prepared by the testing facility. Purchase and processing Page 7 of 65 Exygen Research Page 38 of 119 Interim Report #26 -Analysis of GroundwaterSamples Exygen Study No.: PO000760 Exygen Protocol Number: PoooO760 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. Water, soil, sediment, fish, clam, vegetation, small mammal liver and small mammal serum samples will be received at Exygen directly from 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 availabilityin the field. The total number of samples received and analyzed for each matrix will be documented in the h a l 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 2OC-8OC. 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 frozen at S -10C. Small mammal whole blood samples will be centrifuged in the field at the time of collection and the serum fraction will be used for the study. Small mammal serum will be stored frozen at < -10C. The receipt and processing of the samples will be documented in the final report and raw data associatedwith 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 8 of 65 .- - Exygen Research Page 39 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Exygen F~~tocNoulmber: P0000760 ANALYTCALPROCEDURE-S References: V0001780: V0001781: V0001782: V0001783: V0001784: V0001785: V0001786: "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 Sedimentby LCMSIMS" "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 Vegetationby LCIMSIMS" "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA)in Small Mammal Liver by LCIMSIMS" "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Small Mammal Serum by LCIMSIMS" 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. VERlFICATION 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 pcr sample collected. The bottles will be 500 mL preclcaned SdSpec Premier wide mouth HDPE bottles. These bottleshave been routinely used for PFOA sample collectionat the testing facility and have been shown to be &ee of PFOA. All containers used for water sample collection will be shipped to the sample location containing 100 ng of 13cPFOA (equivalent to 500 ng/L in the final sample). Sampleswill be added to each containerto 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 spkes will contain PFOA as well as perfluorobutanesulfonate (PFBS), pduorohexanesulfonate (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 Page 9 of63 Exygen Research Page 40 of 119 Interim Report #26 -Analysis of GroundwaterSamples . Exygen Study No.: PO000760 Exygcn 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 preclmed ScilSpecPremier wide mouth HDPE bottle per sample collected or a zip-seal bag. All contain&ags 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 JeveJ and processed through the described prucedure to determine method accuracy and to check for bias. "C-PFOA will also be added to each sample in the laboratoryprior to extractionat 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 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 laboratoryprior to extractionat the level specified below. Vegetation Small Mammal Liver Small Mammal Serum 10 ng/g 10 ng/g IO ng/g 100 ng/g 100 ng/g 100 ng/mL 100 ng/g 100 ng/g 100 ng/mL 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 "C PFOA results to establish precision and accuracy of PFOA samples. The following samples are to be prepared: Page IO of 6S --- Exygen Research Page 41 of 119 Interim Report #26 -Analysis of GroundwaterSamples Exygen Study No.: PO000760 Exygcn Protocol Number: POW0760 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 n g 5 with I3CPFOA and at 5000 n g 5 with PFOA. The above samples are to be extracted and analyzed according to method V0001780 entitled "Method of Analysis for the Determination of Pduorooctanoic Acid (PFOA)in Water by LC/MS/MS." METHQD FOR CONTROL OF BIAS Control of bias will be addressedby 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 of 65 Exygen Research Page 42 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Exygen Protocol Number: woo0760 f4 The name and address of the Study Director, Sponsor Representative, and of the testing facility. 0 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 Stwly 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. Descriptionof the instrumentationused and operatingconditions. All results fkom 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 chromatogramsof 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 documentedin the report. Locationswhere raw data and the finalreport are to be archived. Additions or comections to the h a l 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 SponsorRepresentative. SAFETY AND HEALTH Laboratorypersonnelwill 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 Page43 of 119 Interim Report #26 -Analysis of GroundwaterSamples Exygen Study No.: PO000760 Exygcn ProtocolNumbcp:PoooO760 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 amendmentswill 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 documentedand reportedpromptly to the Sponsor Representative. DATA RECORD KIWFlNG Records to be maintained include the following (as appropriate): 8 Sampletracking sheet(@ Sample receipt records. storage history, and chains of custody History and preparationofstandards (stock, fortification, calibration) Descriptionof any modificationsto the method 8 Instrumentrun sheets, bench-sheets or logs 8 Analytical data tables 8 All chromatographicand instrumentalconditions 8 Sample extraction and analysis dates A completelisting of study personnel,signaturesand initials Chronologicalpresentationof all study correspondence Any other documentationnecessary for the reconstruction of the study Chromatograms-All chromatogramswill contain the following: 8 Sample identification, injection date, amlw or other indication of the area of interest, and injectionnumber correspondingto the run. Additionally, fortifications will include the amount of analyte added and the sample number ofthe sample that was fortified. 8 Analytical standard chromatograms will additionally include the concentration (e.g.. pghnL). Page I3 of65 Exygen Research Page 44 of 119 Interim Report #26 -Analysis of GroundwaterSamples Exygen Study No.: PO000760 Exygen Protocol Number: PoooO760 As part of the documentationthe 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 792.185. QUALITY ASSURANCE The QA Unit of Exygen Research will inspect the study at intervals adequate to assure compliancewith GLP's, and will report the findings of audits to the StudyDirector, Exygen Management, and the SponsorRepresentative. RETENTION OF DATA AND ARCHMNG 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 informationh m 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 from the study director before discarding or returning samples. Exygen Research Page I4 of6S . - Page 45 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 APPENDIX I ANALYTICAL METHODS V0001780 "Method of Analysis for the D e t e r m i don o. A ;rfluorooctanoic Acid (PFOA) in Water by LC/MS/MS" V0001781: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS" V0001782: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Sedimentby LC/MS/MS" V0001783: "Method of Analysis for the Determination of Pduorooctanoic Acid (PFOA)inFish and Clams by LC/MS/MS" P- V0001784 A"Mciedth(PodFOofAA) innaVlyesgisetfaotriotnhebyDLetCer/MmiSn/aMtiSon" of Perfluorooctanoic V0001785: "Method of Analysis for the Determination of Pduorooctanoic Acid (PFOA) in Small Mammal Liver by LC/MS/MS" V0001786: "Method of Analysis for the Determination of Pduorooctanoic Acid (PFOA) in SmallMammal Serum by X/MS/MS" Exygen Research Page IS of 6s Page 46 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 Exygen Research Page I6 of 6s - -- ..-- Page 47 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.:PO000760 Exygen Protocol Number: PO000760 wm- W N m b r VOOOllM I I MALITICAL METHOD MahDd of Anrlydc formS DaunmiDpionof PCrBuoroocutloicAcid (PFOA)in Water by LCIMSIMS 1 .o 2.0 3.0 4.0 5.0 Exygen Research Page I 7 of 65 - Page 48 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 T3mhinlu 0.0 65 1 .o 65 8.0 25 20.0 25 22.5 65 PloWlCo u&lJKmm 35 0.3 35 0.3 7s 0.3 75 0.3 35 0.3 - 6.6 InjgjcccionVohunc: 15 pL (canbe iapaudto Y much u 50 gL) 6.7 puntitltioll:PolLAm caerml atandudulibntioncurve. 6.1 ~ u o ~ i m-s23: mi~~te1. Tbo.boMsonditiQuMinradodu I guide udndrmy bo changedm order to optimiu(bOHpLC1yWm 7.0 M s m d s S F 7.1 MEloctrmpnr Noptivo MRM modo. monitoring413 -b 369 m/z. Tbc above ColditiOnr M in(Crd0d u I &la .ad may be changed in orda io cp4imka tho MSMS 8ystem 8.0 FTcpuuhofbl* 8.1 MobilePhw 8.1.1 2 mM d m rct.tc in wtex u prop.rod by d i n g O.IS4 y of M m o d p m ~ t 1oOOOmLofwum. Al~W~MYbopnplrcd Exygen Research Page 18 of 65 Page 49 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Exygcn Protocol Number: PoooO760 Exypheueb MdmdI'hhrVW1780 I 1 ANALYTICAL METHOD MabodofAnrlylisforb DctcrminuiollofPatluomocunok Acid (PFOA) in Wuer by LCIMSIMS 9.1 SundpdStadr/portifiutiollSoutiOn 9.1.1 R c p r o I d a L ~ n o f - I W p ~ o f P F O A b y w a1~0mpg of dyt*.lundud (mrrcctcd for purity) md dilute to 100 mL with mollvnoIin.12~mL~LDPEbotllc. 9.1.2 A lOpc/mLhtuhaon whxion of PFOA is p m p d by brinlyng I O mLoftho100p~wlutionO.fuulvolumcoflwwithmclh.Ml in I 125 OIL LDPE boltlo. 9.1.3 A I . O p p h n L l o r c i f ~ . t i o n w l u t i m o f P P O A i s ~ ~ b y b r i n g i1n0g mL of m0 10 p- lolution to I find volume of 100 with mcihMol in I 125 mL LDPE bonk. 9.1.4 A 0.1 &nL fonifkalion w l u t i ~of PFOA is p r e p d by bnnging I U mLofthol.Opc/mLwlutionaifuulvohvncof 100 withmrlunolin rlZ5mLLDPEbotlla 9.1.5 A 0.01 pghL fintifiraim wlution of PPOA is p l o p d by bringiny 10 mL of h e 0.1 pc/mL lolulim to 8 hl volume of 100 with rm(hm1m1 I U m L L D P E W a 9.1.6 T&#OI& d fm(i5ution mlutiolv.IC lo be dorod inI rcbgoruor II eppoxbutely4% urd Mstable for I maximum period of 6 months fmm tho &to of plopmtion. 9.2 Suldrrdc.libntionSolulioru 92.1 LOMWMS u l i o n rtlndvdt M pnpuod in HPU: w u u . the u l i dmdud~yc poccucdtluuugb thc cxtnction proccdurc. identical to umplor 93.2 Tbo IolloviDpi8 I typiul exotpla: additional coI1cdnlntionsmiy bc Exygen Research Page 19 of 65 Page 50 of 1 19 Interim Report #26 -Analysis of GroundwaterSamples Exygen Study No.: PO000760 Exygen Protocol Number: PoooO760 12.1 12.2 12.3 12.4 Exygen Research Page 20 of65 Page 51 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen StudyNo.:PO000760 Exyeen Protocol Number: P0000760 13.0 13.1 13.2 133 13.4 13.5 13.6 14.0 Exygen Research Page 21 of 65 -- Page 52 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.:PO000760 Exygen Protocol Numbrr: PoooO760 Exygen Research Page 22 of 65 Page 53 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.:PO000760 Exygen Protocol Number: PO000760 ANALYTICAL METHOD M a b o d M ~ m kVOW178l ww- 3058 W h Drive Stam Coll.ea PA 16801 SOU by Exygen Research ToULPyol: 7 Page 23 of 65 ____. _. Page 54 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Exygcn Protocol Numkr: PoooO760 1 .o 2.0 3.0 4.0 5.0 Exygen Research Page 24 of 65 . Page 55 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Exygcn Protocol Number: PO000760 bs-- UnLOdNrmbr VwO1781 I 1 ANALYTICALMETHOD M W o f W & tiuoptluIkmmidm of PaflumraevlDicAcid (PFOA)in Soil by LCIMSIMS 5.14 W ~ ~ I I ~ ~ . 5. I5 C d h cupablc of@inning50 mL polypmpylenctukr at SO00 rpm. 6.0 C h m r m t O ~ S y r t a m 6.1 AMI@WI Columa:Fluophc RP (Kcmonosoioati6c). 2.1 mm x 50 mm. %I (Pm 825M452130) 6.2 T y 30% 6.3 Mobile Phvo (A) : 2 mM Anrmaaillm Awtata in Watm 6.4 MobileFiuw(e): Mclb.nol 6.5 G m l i ~ U P m g ~ : Thmaialy 0.0 1 .o 65 8.0 25 20.0 25 22.5 65 Fbw Rae &RfInKIiw 35 0.3 35 0.3 75 0.3 15 0.3 35 0.3 rc Exygen Research Page 25 of6S Page 56 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Exygcn Protocol Numkr: PoooO760 9.0 Surdud- 9.1 Stlld.rdStn&Fd&.tioll Solution 9.I. I Rcp.rc a stack solutionof -1 W pdmL of PFOA by weighing IO mg of rnrl- *mdpd(oonostedfapluily) and diluta to I00 mL with maimol in a 1u-am p e bottls. 9.1.2 A 10 )I&&fd6UtiOnwlulion of PFOA is prcpurd by bringing IO mL of tk 100 p&& solution to a find volume of 100 with mlhnol in a lu mLLDPEboala. 9.1.3 A 1.0 pplmL fntifiwiollsolutionof PFOA isprrpucdby bringing IO mL oftho 10 pdmL solutionto a find volume of 100with ndunol in a 125 mLLoPE 9.1.4 A0.1 p&&fomticlMn ~Iolutionof PFOA u pnpuodby bringing IO mLof tho 1.O p&& wlutiocl to a 6 d volume of 100with mdunol in a 125 mL LDPE boala 9.1.5 A 0.01 p#mL fdficdim solutioa of PFOA is p r c p d by bringing IO mL of tho 0.1 pJmL solution to a R d volume of 100 with mab.llolin L iu m~mpebotua. 9.1.6 Tho mock and lorcl5cdion~IvCiOnrw to be s W in a refripasator u rpporimrtclyIT and w stable for a nuximum paid of 6 monthr 6mmthodaaofprcQuuioa 9.2 SbndudC!diion Soluliau 9.2.1 933 LclMslMs ulik.tioa 110 pnpuod in HPLC wuas. The dibntiou asdub m psoccuod thmugb the nrmction procedure. idddtollmplor Ibcfollowing ia typid ~ ~ p n paldd~it:ianrl concco~ioncmay be PrapMdunsCdsd. Find &wnbuim Fmiflclcioa Volwwof Conwmimof ofFmifiarm Volume Fa~iif*dcocuml Wibnlim Calibntion SundudID SOluIirmW) OL) 0 0 IO 100 IO 100 SMdC(mL) 40 40 40 Sludrdlwtl* 0 2.5 50 (ex.mulC) XCmmddyy.0 xcmyy.1 Xcmmddyy-2 IO 4m 40 100 xcmmddyy-3 im 100 40 250 xcmmddyy4 100 100 40 500 XCmmddyy-5 -100 .Ihonmqrd- . 400 40 loo0 xcmmddyy 4 o f t k u l i o n atmdmdisqual to 8x ib initial ~ ~ W ' t ofthaamdhorld m o smrction~ (SPE). xc oxmorod uubntbasmndad. Page 26 of 65 Exygen Research Page 57 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.:PO000760 Exygen protocol Number: PoooO760 Exygen Research Page 27 of 65 ._- Page 58 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 gwv.- I Methodof Anal* M.cLodN h r VmO1711 I AIyALmcAL m i l O D lortbe D m d n U i o n ofpclnuomo*.noic Acid PFOA) in Soil by LCIMSIMS 13.0 Aacpt.lroClit& 13.1 Cbmutommudlbow a pmk ofada&Ier ion U 369 m u Rom a puenl of 413 am.'Ib.413 ~u pMu CUNQOI& to tba PFOA anion. while Le aurghmim(369Mu)rrpclcntltho lou of urbondioxide. 13.2 Mabod bhnbmudod W PFOA U b o b grcu8rU1~t1he LOQ. If a blank codm PFOA U Imb proltathm50ngA, them a l l ~ wblank umplc mwtkobt.iaodndtho~rrtinvtmwbcrocrrmnPd 13.3 ~ O S O f o O ~ ~ Q i k c r M d d Q ~ ~ ~ b o b70a-13wO.hcOcf n b i r knownvalua. If a m8ol &ike fill8 outddo tho .cccpuble limits. the cntim (d of umpla diould be n-.xlnctcd. Any matrix rpike outside 70- 130% should bo cVrhu(0d by the uulylt to daamine if rc-exlnction is WPRlltLd 13.4 Any d k d i c a rurdvd fomd to be a shtkicd ouUior by ruing the Huge Enw Tat. m y be ~ c l u d o dfrom tho ularluion of the calibration curve. H ~ ~ w ehtrm, wl n u m k of Qmcrod dbntion ItMdudr that muld be ududod must mt cxwod 20% of tba total number of cxmftcd rtudardr iqjecbd. 13.5 l l m Eonoldion EwffioieM (R) for ulibntiw c w o l gc& muat be 20.9% (R' 20.985). If lion rclultt fa11 o v u i d ~l b o r ~limita. hi .pproprilto *cp mutt bo llLoa to &jut iDmumau opntion. and the It.nd.rdrorth m h t yt ofumpla rhouldkwdyrod. 13.6 Ratmh'on timol bdweua d u m p l a mutt not driA mom Ih.n f 4 Kwitbinm~arlyricll m UraoDtlon timadriftexcocd~tbir limit within .IImulyUcal n m t b t~ho nt m u t k nuulyzSa. Exygen Research Page 28 of 65 - Page 59 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Exygcn Protocol Number: PO000760 14.3 UICIhfollowingopvltion lo wnvm tbs munI of PFOA found in n f l lo rulp W). PFOA found (pp-b) * SmIlplcweight(5 g) 14.4 Un tho f o l h i q qwtioll to ul&c bUCdOlldryW6igllL tb mount of PFOA found in ppb PFOAfacrd @pb) dry wcigk =PFOA found@pb)x [100%/ toul rcliWN)] Exygen Research Page 29 of6S -- .- Page 60 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 ANALYTICAL. METHOD M a h o d N e V0001782 L Due Exygen Research ToulPysr: 7 Page 30 of 6s Page 61 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Exygen protocol Number: PO000760 Exygen Research Page 31 of 6s .-. Page 62 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.:PO000760 Exygcnh t o c o l Number: PoooO760 6.0 IidJnbI 0.0 1 .a 8.0 10.0 22.5 %A. &R 65 35 65 35 25 75 25 75 65 35 ... . . Imllmin) 0.3 0.3 0.3 0.3 0.3 7.0 8.0 Exygen Research Page 32 of 65 Page 63 of 119 Interim Report #26 - Analysis of GroundwaterSamples Exygen StudyNo.:PO000760 Exygen Protocol Number: PO000760 Wm- Ld.ILodN u d m VOml782 I i ANAL.Y"lCAL METHOD Mabod of M y d iforb Dotaminr6n,ofPerflmuoroocuaoicAcidQFOA) in Sodimen1by LCIMSIMS 9.0 9.1 S t a r I d u d S ~ ~ S o l v l i o n 9.1.1 R0p.na sIo& colu(i0n of -100 pghnL of PFOA by wmghmg IO m8 of llulylicrl amdud (conscodfor purity) midilute to 100 mL with malmol in 1 115-mL LDPE Me. 9.1.1 A 10 p a f d f i d o n colution of PFOA u propndby bringing I O mL of clu 100 pphaL coltnirmto 1 Bnrl volumo of 100 with methanol in a 125 mL LDPE bottle. 9.1.3 A 1.0p@ Bntifiution mluhnof PPOA i PrrpMdby bringing I O d o l tlw 10 p@ colution to 1 A d volvmc of 100 with mthnol in 125 mLLDPEbotlle. 9.1.4 A 0.1 tostilldon mlutionof PFOA 11 ppadbybringing 10 . mLof Uu 1.0 p@ colutionto 8 ~IU voluImo of LOO with m t h l m a1UmLLDPE~. 9.1.5 A 0.01 p@ fomhlum colutbn of PFOA is pleplrrd by bringing 10 d Of tbo 0.1 p@L lohhl to 1 Ibd V O ~ u m COf IM) Wlth nr(b.adin. 125 mL LDPEbotllo. 9.1.6 Tbc ltodr ad forti5o.tioncoluliom Mto be norodin a rcfrigmtor 11 -y 4'C ad M stab10 for a muhum poriodof 6 months fnmclu dueof plcp.ntioo. 9 2 sclndpdt 3 b b d o n Solutirm 92.1 Lc/Ms/Msulibntimruldpdr 110 prop.rod in mcuvaOl via dilution 0fcluO.l (rphaLblu6cuimWlUtiaL 9 2 3 llw followingu 1 typiuloxlmpk dditid SOMlltruions may be WcrUndUIlWkd. FilNl ofFmihth V h solulh(ad.IL) (mL) Dihpdto (mL) Caoca~UUion (D&L) 100 IO 100 10.0 100 5 I00 5 a 100 2 100 2.0 IO IO 100 1 .o 5 IO 100 0.5 -.. .... Exygen Research Page 33 of45 Page 64 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Exygen Rotocol Number: PoooO760 wI-- MnLod Nu&a V00011t2 I I ANALYITCALMETIIOD Mabod of And@# for tho -nation ofPcrfluorooctmoicAcid (PPOA) in Sedimentby Ll3MSfMS . 9.2.3 Stam dl ulibntion dmdarda in IZSmL LDPE narrow-mouthbollls atZ*Cto6T,~ptodxm0ntl~ 9.2.4 Altormtevolumm d COWlltlltlOlU oflt.ldub my k propMd u mdod. 10.0 Bltchsct Up 10.1 Euh b.lcb of lrmpla oxlnctd (Wully 20 or lea) mult include m l u s t 0110 mtmtad c a m 1 ad (WQ uahor(0d CMhDl: fonifid u known -usion# (lab conIrol #pike)to Val& pmculunl recoveryfar the balfh. 10.2 Rot, ' . for dcld ud L.bontory duplicuu ad pika will ba rpccilied in tho q d t y lyvllllcop h forthia project. 11.1 11.2 11.3 11.4 11.5 11.6 11.7 11.8 11.9 11.10 11.11 11.12 11.13 - ._._.- Exygen Research Page 34 of 6s Page 65 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 12.1 12.2 12.3 12.4 12.5 13.1 13.2 13.3 13.4 13.5 Exygen Research -. ..... . . __- Page 35 of 6s ._ Page 66 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 ExygcnProtocolNumber: PO00760 14.0 CI*u*tiorn 14.3 UMtbo r o lcq~don to follvaUm yllwlllof PFOA found in nm lo 4 8 @Pb). Exygen Research Page 36of65 ._ -- Page 67 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.:PO000760 Exygen Protocol Number: PoooO760 ANALYTICAL METHOD MduMNuuhx VooO1783 .. ... Exygen Research Page 37of 6S Page 68 of 119 ~ ~~ Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Exygy Pmtocol Number: PO000760 'wmlloou* Makd of AMI* 1.0 himdndN~VOW1711 AlyALyTlcAL METHOD forthe IMedmtirnof Porfhnooavpic Acid (PFOA)in Fish and clulubyLcIMsIMs 2.0 3.0 4.0 5.0 Exygen Research Page 38 of 63 Page 69 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen StudyNo.: PO000760 Exygen Protocol Number:PO000760 5.4 5.5 5.6 5.7 5.8 5.9 5.10 5.11 5.12 5.13 5.14 5.15 5.16 6.0 ChmnutogqhkSwtmn 6.1 Adylicd Cohunn:PlUDphuc RP nrcyMIc Siiontiflc).2. I nunx 50 mm. 5p ( P N 8Z505-052130) - tf 6.2 Tea~pmme:3W: 6.3 MobihPluM(A): 2mMAmmoniumA.c&UoinWUa 6.4 MobiloPhw(B): Mah.001 6.5 Gr8dimtPmm: now Rue %BhL!minl 0.0 35 0.3 I .o 65 35 0.3 8.0 7.5 75 0.3 20.0 25 75 0.3 22.5 65 35 0.3 6.6 Iqialioa Volume 15 pL (cank iacroucd(0u much u 50 KL). 6.7 Qumtiwh: Pcsk A m - oxtnvl Itnbrdulibntion curve. 6.8 Runw-Umirmtar. Tbs .bow saaditiolv Mintdod u L &do ud may be dunged in order to opamizouuHPLclyltom. Exygen Research Page 39 of 65 Page 70 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Exygen Protocol Nuder: PoooO760 7.0 MSlMSSyllom 7.1 Modr: Blwbolpny Nepciw MRMmodo. moniming413 -b 369 m/z for PFOA. The abovecolditiolllyo intcrdtdas iplidc ud m y be clungcd in order to OplhiZehMSMS~ 8.0 ~ o n o f s o l p t i o l l l 8.1 Mobikphuo 8.1.1 2 mM d u m rotlto in w r t a is PrOpMd by adding 0.154 g of mnmmillmscam to loo0 mL ofmta. 8.2 exh.e(ionsolutiolu 8.2.1 2% ucorbic loid in methDolUpCpMd by d * l o l v i ~2 g of ascorbic ridinlWmLofmahuto1. 8.2.2 30% D ~ y l ~ in to~lumwais pn~pMcd by bringing 3 mL o f ~ y ~ to ifhfnrlvoelumeof IO mLwith tolume. AltCiaWVOIUUlMUUykpopmd. 9.0 Sullaod- 9.1 sMc*rdslocwpatiflutionsowion 9.1.1 Ropuoiltoolr loldon of-100 pghL ofPFOA by weighing 10 mg of d y l i u l Usdad (Hmostod f a purity) and dilutc to I 0 0 mL with AI in a 1 2 s - mm~ p e ~OIUC. 9.12 A 1.0 p#mL forti6catk.n solutionof PFOA u pcp.ndby bnnging I m. mL of tho 100 p&IL solution to iBrvl vohunc of 100 with mcthmol in a 12s mLLDpB 9.1.3 A 0.1pdmLf d 5 d m loWmof PFOA & PlpIlSd by bringins 10 mLof tho 1.0 p&IL solutionto if h l volumc of 100with mcdunol in ii u m~m eboala 9.1.4 A 0.01 pg/mL fortifidon solution of PFOA is PlOpMd by brinliny 10 mL of tho 0.1 &mL solution to a Brvl volume of 100 with .*clu;lolin 125 m~m p e boato. 9.13 Iho mck Od foiti5caion aol~~tioM m to bo atmodin a reffigentor11 t~polirmtdy4~uCd M rtlbla for a nuximumperiod of 6 mon~hr h hdateof plopultiw. Exygen Research .. . - Page 40 of 65 Page 71 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Exygen Protocol Numbcr: PoooO760 P S O M i m W ) (mL) (mL) 1 .o 5.0 I00 OlW) 0.05 1.0 2.5 100 1 .o 1.o I00 0.025 0.01 0.05 IO 100 0.005 0.025 10 I00 0.W25 0. I 10 100 0.001 0.005 100 0.m5 93.3 SaPnJl C a I i b L l Illldud, in 125-mL LDPE nurow-mouth M e 3 m2.cto 6.c. up to dx mtiu. 9.2.4 Altan#te Voluma aod c u ~ ~ ~ ~ t r u oifoslmvrdad~may be prrpvodu medad. 10.0 W W l J P 10.1 Bub buchof sanpla aanaod (typically 20 or leu) mud include 81 leut ow lmhapd canhol mld hro lmhaod Coeholl dMifiod ll known COKOlYntiOlll(lab Csn(r0l c p h ) to Vaiflpooodunl rccovary for rho buch. 10.1 R ~ ~ ~ ~ I W OfarM5&U ad Lbontory duplicsta and s p h will k specified in tho qualityu s u m c o p h for tbii project. 11.0 SmnpkExwim 11.1 Wei@ 5 g of bzm wupb into 50 mL polyempylonc catrirUgo tubes (lorti* u lloodrdrop*oolidmdmix well). 11.2 Add 30 mL ofawbmibiib aod OD a m * 1 adm h k o r for-15 minuter. 11.3 P l w t b o @ b a i n a ~ f o r - l b r u . 11.4 P d aod coldibolltho SPEblbcl .adrilvlizc (bo par-- flukr. 11.5 Pack tbo 10mLSPEtabe1 h lopuacc with 2 g florid.2 g riliu gol. 2 y cuboa, aod 1 g LC-NHz. Conditirm (bo colvmru with 20 mL of mcthml. tboa 20 mL of leaordbilc. oirud all wuhcl. Do not dlow h e column to dry. 11.6 S i h h tbr 1zT mL pouJbscd Iluka by rinsing with Ute 30% dimdtlyldichbmdlnoin tolucno Iolutiw. Rinr ch fluLwith tolusllDonce, fallowed by meUunol (URW h)t. hy tbo !luhcomplotcly before ue. dthrby lldryllllorwithastmaa ofniwogm. Exygen Research Page 41 of 65 -- Page 72 of 119 Interim Report #26-Analysis of GroundwaterSamples Exygen StudyNo.: PO000760 Exygen Protocol Number: PoooO760 11.7 11.0 11.9 11.10 11.11 11.12 11.13 11.14 11.15 11.16 11.17 11.18 12.0 c l u a m u o m y 12.1 12.2 12.3 12.4 Exygen Research Page 42 of 65 -_ __ -. Page 73 of 119 Interim Report #26 -Analysis of GroundwaterSamples Exygen Study No.: PO000760 Exygen Protocol Numbcr: PoooO760 .- - I I ANALYTICAL METAOD Mabod of Arulyds tor Ih.DaamLvtonof FwhomffuoOic Acid (PFOA) in Firh ud C*nubyLc/Ms/Ms 13.1 133 13.3 13.4 13.5 13.6 14.1 14.2 Exygen Research Page 43 of 65 Page 74 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.:PO000760 Exygen Protocol Number: PoooO760 - 14.3 For umpk lortifind witb known mom48 of PFOA prior u) extraction. UIC thc followillgOpdmlo cdcul.tc thc paccnlIccovay. Recovmy (%) Exygen Research Page 44 of 65 - Page 75 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.:PO000760 Exygen Protocol Number: PO000760 Exygen Research Page 45 of 65 Page 76 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Exygcn Protocol Numbcr: PoooO760 1 .o 2.0 3.0 4.0 5.0 Exygen Research Page 46 of 65 Page 77 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Exygcn Rotocol Number: PO000760 I Mehodof f a tho I AlyALYTlCU METROD ofPufiuomoeImoic Acid (PMA) in Vegnrlion by LCIMSIMS 6.0 chmm*ogrplu'E spml 6.1 Ao8lytbI Column:Ruophuc RP (KcyUom SOionlifis). 2.1mm x 50mm. Sp (PN 112505-052130) - % 63 Taqmture:3OY: 6.3 Mobilepboo(A) : 2 mM AmmDnium M 6.4 MDbibPho(B): Mctbm~ol 6.5 GndicntRogun: e in W.ra Plow Ruc kLB6nuninl 0.0 35 0.3 I .o 65 35 0.3 8.0 25 75 0.3 20.0 25 75 0.3 22.5 65 35 0.3 7.0 Msm(9Syltom 7.1 Made: BI&bolpnyNe#ativr MRM modo, monitoriq413 -D 369mlz for PFOA. Exygen Research Page 47 of 65 -. -_ Page 78 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen StudyNo.:PO000760 Exygcn Rotocol Numbcr: PoooO760 ?be above owditiaum intadsdua guideand m ybo clungod in order to OgtiminothcMSMs~. 9.1.1 9.I.2 9.1.3 9.1.4 9.1.5 Rcpuc a stock mlutimof -100 &nL of PFOA by weighin&I O mg of rmlyt*.l stmldmd (eorrocld lor purity) and dilute 10 IW mL with msttvnolio1 17.5-mLLDPBbottle. A 1.0 fonifiuciOa solution of PFOA u pnprrod by bringing I mL of tb 100 @mL solution 10 I dnrl volume of 100 with m c h l io a 123 mL LDPEbatlo. A 0.1 p&tL hti8ulionloltuionof PFOA u PICpMdby brinsing 10 mLoftbD 1.0p#mL solutioato 1dnrl volumeof 100with mthnolin a 115mLLDPHboWa A 0.01 &mL f&li5clrion solutionof PFOA ICPrqHndby bringing 10 mL of tb 0.1 flmmlutioll to 1 Brvl volume of 100 with nmnmwlin a 17.5mLLDPEW e . Tholtalrlld dmifiutioa mlutiolum to be rtorodin a mbipator at mppxhuely 4 T .Idm stable f o r i m u h u m period of 6 months ~1&~OfprsprUion. 9.2 SmadudCllibn(ionSolutiorv 9.2.1 LclMslMsdWcnItladub PO prrp.rcdin m*h.nol via dilution of tho 1.0 p&hlLthti8crionsolUtiaL Page 48 of 65 Exygen Research Page 79 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 9.23 Thr following ia a weal example:dditiond concamu'OIU may k prrp.rd==kd Find ofFatihh Vohn Dilubdro Conoamam solulim(rzhnL) (d) (mL) (lldml.) 1.0 5.0 100 I .o 1.3 100 I .o 1.0 I00 0.0s IO IW 0.0s 0.025 0.01 0.003 0.025 IO I.0.0- 0.0023 0. I IO 100 0.001 0.003 IO 100 o.Oo05 9.2.3 Slon JIe d h l i o a Nldrbin 125-mL LDPE numw-mouth bottler urCto6.C.UD to dx monmC. 11.1 11.2 11.3 11.4 11.5 11.6 11.7 Exygen Research Page 49 of 6s Page 80 of 119 Interim Report #26 - Analysis of GroundwaterSamples Exygen Study No.: PO000760 Exygen Protocol Number: Po000760 12.0 avormtoorphy 13.1 clnmvtopmmu# cbow I pmk of I duybca ion .t 369 mu fmm Ipuenl of 413 OIL 'Ibs 413 .mu pra c m q o o d #to tbo PFOA mioq while rhc dw@uion(369mu)rop~lolmcbo lor,of wboa dioxide. Exygen Research Page 50 of65 .- .Page 81 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.:PO000760 Exygen Protocol Number: PO000760 13.2 13.3 13.4 13.5 13.6 14.0 l3kuMom 14.1 F 14.2 14.3 Exygen Research Page 51 of 65 Page 82 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.:PO000760 Exygen Rotocol Number: PoooO760 Exygen Research ToQlPya: 7 Page 5.2 of 6J Page 83 of 119 Interim Report #26 -Analysis of GroundwaterSamples Exygen Study No.: PO000760 Exygcn Protocol Number: PoooO760 ' 1 aim- MdbdNuabu VOW1795 WALWICAL METHOD Mabod ofADllydr forthe DctmninUimOfPorfluomoaPIoicAcid (PPOA) in Small Mmmad Liw by Lc/Ms/Ms I .o 2.0 3.0 4.0 5.0 Exygen Research Page 53 of6S . -___ Page 84 of 119 Interim Report #26 -Analysis of GroundwaterSamples Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 5.9 Diqnmbbm 5.10 ~~(lM)-lMW)~md10-1W(rL),withdirpoorblctipr. 5.11 Wmm Scp Pak Vlo 6 oc (I& tCl8 SPEcutridpu. 5.12 SPE vacuum nunifold. 5.13 T i . 5.14 Wrirr-Utianlhkor. 5.15 catrifugacp.bleofapipirminl15 mLpolypropyla~tubes m 3WOrpm. 6.0 ChmnutogrmphkS y U m 6.1 Anal- colrmm:Fluop&#RP (Koyrtooo Scioolific).2.1 mm I 50 mm. Sp (PN 8ZSOEo52130) 6.2 Tmpmhne: 30'C 6.3 M&ikphSc(A): 2mMAmmmivmAfctucinWItcr 6.4 Mobibpbuo(B): Mab.nol 6.5 O n d i a U R o ~ : nm&W%A 0.0 65 I .o 65 8.0 25 20.0 25 22.5 65 Plow Ruc " Y 35 35 0.3 75 0.3 75 0.3 15 ' 0.3 6.6 6.7 Inj& V o Qluotirrtioa: P k uk I5 @ AIU -(CUI ex bo ta MIo u much u d uadudcllibntionw 50 e pL). . 6.8 RunTi -23minUta lblbovo condition,ua kuordodu .guide mdm y bo clungedinordcr Io optimiz8r&HpLc~ 7.0 MSIMSSWcm 7.1 Modo: ElecbwpayNegalive MRMmodc.m~llitming413 -b 369 m/z for PPOA. lb lbow CaditiODl IIintdsd u iguide md m y bo faangod in order to opthizer&MsMs~ P Exygen Research Page 54 of 65 ~. Page 85 of 119 Interim Report #26 -Analysis of GroundwaterSamples Exygen Study No.: PO000760 Exygen Protocol Number: PoooO760 Exygen Research Page 55 of 65 Page 86 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.:PO000760 Exygen Protocol Number: PoooO760 10.0 BatchSdUp 10.1 Brb batch of umpk o)(tTytod(typically 20 or leas) muat include PI least one ullpcltcd Eoluml Isd two Mmmla fortified at known (lab conkelrpiko) to Wriflprocodu~I~OCOYQfYor the batch. 10.2 R o q u i ~ m m bfor field ud *bonrmyduplicuol .ad@ea will be spined in thcquality.IIIIIWp l for~ tbia pjkt. 11.1 112 I13 11.4 11.5 11.6 11.7 11.1 11.9 11.10 11.11 12.0 chmmrto*y 12.1 12.2 123 12.4 Exygen Research Page 56 of 6s . - . Page 87 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 13.1 13.2 13.3 13.4 133 13.6 Exygen Research Page 57 of65 Page 88 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Exygen Rotocol Number: PoooO760 %mwh ~~vOoo17aJ t I ,wwric,uMETHOD MCtbodofAnrlyshIbrbothoDmmidonofPduMmctmoic Acid (PFOA) in Small M m n ~L~ivler by Lc/Ms/Ms 14.2 For llmptr Iortiilod wilh known MMlDy of PFOA prior lo cxmction, use tho fohwiog eqlutian lo u*ul.to UW pucQlt rocovay. 14.3 Exygen Research Page 58 of 65 Page 89 of 119 Interim Report #26 - Analysis of GroundwaterSamples Exygen Study No.: PO000760 ExygenProtocol Number: PoooO760 ANAL.YnCAL METHOD MabodNumba: VOOO1786 Exygen Research TdPlgor: 7 . .. Page 59 of 6s ___. . Page 90 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen StudyNo.:PO000760 Exygen Protocol Numbcr: PoooO760 Exygen Research __ -. Page 60of 65 Page 91 of 119 Interim Report #26 - Analysis of GroundwaterSamples Exygen Study No.: PO000760 I - 1 hYa= MSlhodNvnbrr VwPl 7Ob .uxAitync.uMETHOD Mahod of Arvlylir for Ilw Drtaminuion of PerfluarowUmic Acid (PFOA)in Small MamudSonunby LClMSlMS _. 5.14 W n i - d m - . 5.15 ~ ~ ~ ~ ~ O O f I p ~ 1 ~ ~ p o l y p r n p ~ ~ N b o l a t 3 ~ r p m . 6.0 C b ~ ~ ~ I ~ g r a p l i i c S y r t o m 6.1 An8I*cd cohnaa:plvophwRP (Keytone Scientific).2.1 mm x 50 mm. Sp (PN 82505452130) 6.2 TanpUum 3 0 T 6.3 k b d e pbvo (A): 2 mMAmmonium Acot.ta in Water 6.4 Mobilephus(B): M a h a o l 6.5 ondieorProw: ROWW l k l m i n l F y * 0.3 0.0 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 6.1 6.8 - - lnjcstimVolume: 15 pL (uobo iaaowdto u much u 50 pL). Qaa~tituionP: rrlcAwn wtaual rtlndud cdibruioncurve. Rm Time: 23 minucsl. Ths above condition#Minvododu iguide od m y bo changed inordn lo opttnizotho Hpu:iymm. 7.0 Ms/MsSyUarr 7.1 M a BlctmrpnyNoptivoMRMmods.moni~413~369m/zfor PFOA Tho above d c i o l u M intadcd u a plida and m y bo changed in order to opthniza mS M S M iyrtom. 8.0 RcprntimofSolutioDl 8.1 Mobibphua 1.1.1 2 mM ammonium wmto in w . t o ~i i prepared by dding 0.154 g of d m Y M c 1 0 1OOOmLofwatcr. Altmut8 vohunorm y bo preplmd. Page 61 of 6s Exygen Research Page 92 of 119 Interim Report #26 - Analysis of GroundwaterSamples Exygen Study No.: PO000760 Exygen Protocol Number: PoooO760 am- M*IpdMvllbu VWO178b I I ANALYTICAL .WTIIOD Mclbod of A d l y l i D for tho k h n h s t i o n ofPeffluDmoclMoicAcid (PFOA) in Small M.mmrlSorumbvLCIMS/MS 9.0 st.ndpdRcpmtion 9.1 SIladvd SlofWPomeulion Solution 9.1.1 RopM aock mlution of-I00 fi&/mLof PFOA by wci@ing 10 mg of mrlyriulctmdud (COnMod for purity) ud dilute u, 100 mL with mccha~ilrr 1 izs-cn~LDPE bottle. 9.1.2 A 1.0&mL f o r r i f i c h mlution of PFOA is prepared by bringinp I mL of tbr I 0 0 fiVmLmlvtionto I Bnsl volume of 100 with mhinol inal2SmLLDPEbouk. 9.1.3 A 0.1 &nL foitificltionmluticaof PMA is preppodby bringing IO mL of tbo 1.0 pglolL mlutiOn to a hrulvolume of 100with mcilunol in 1 12s mL LDPE boltlc. 9.1.4 Tiw st& ad fonificuioa mlolutioolam to bo s t o d in a refrigenlorat appxiuutsly 4 T d M stable for a nuximum period of 6 months franmc dueof prpultim. 9.2 ScodudcllibntionSolurioru 9.2.1 Lc/Ms/MsOllibnrioll smulanhmpslwrodinmaUunolvia dilution of thr0.1 @L.fod5mtionlolulion. 9.2.2 Tho foliowing is typiul clunplo: dditiod CoIlCclltntiona m y be pop.rodunrded. cmr+nntioll ofFariRclth V o l w Diluudto Final Concentrstion solution(nJmL) (mL) (mL) (ndml,) 100 5.0 100 5.0 I00 2.0 IW 2.0 100 1 .o 100 I .o s_a._ IO I.0..0 0..5. 2.0 10 I00 0.2 1 .o 10 100 0.I 93.3 ston dl ulibruica itlld.rdiin 125-mL LDPE m w - m o u t h bottler 10.0 Buchsctup 10.1 E.ch buchof ampla or& (typiully 20 or leu) m w include u least ora uumwd amtrol ad two unhrrtod conbula foniflcd i t known DoIloatntio~(l lambdspike) to vripV procedunlrrsovcryfor the batch 10.2 i b q h m a m fw field and laboratory duplicate# and spikes will be spaifid intbo qualityumnnccplanlor tthpja. Par 4 of 7 Page 62 of 65 Exygen Research Page 93 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.:PO000760 Exygen Protocol Number: PoooO760 t ' M e h d ofAnd+ ANALYTICAL METBOD fur tIm Docaminuiooof PduomoamOie Acid (PFOA) in Small MMnvl Saum by Lc/Ms/MS 11.0 Sunplefimnim 11.1 I 1.2 11.3 11.4 11.5 11.6 11.7 11.8 11.9 11.10 I 1.1I Mcwco 1 m L o f u m p l o ~ a 5 0 m L p o l y p m p y l a a c o l t r i ~ e ~ ~ ( f o n i f y u ar+dcd.mplrc lid ad mix well). Notc Uut altmute volumu of serum may be meuurcd ca theumple I*C avihble foruse. MdsNatotbcumpk fora w voiunw of20 m ~c.lptightly Vorra31for-I T ~ m m k1 mL. of tba umple wing dkponbk pipem into a I5 mL dirpovbloolhifuge tube. M d 5 mLof d b i l e ad dukefor -20 minuteaon a mist-ution shaker C d h p t b a tube8 lt-3OOO Ipmf o r d minutor. Doclnt the supamta into a 50 mL dirpoublc ocrraifuge be ud add 35 mLorwucr. Cmdilim tba CII SPE umidga (I & 6 mL) by p w 10 mL methanol wm). followadby 5mLof HPU:water (- 2 Do not In column Nn dry Lod tba umpk 011 soaditioncdC,I WE c d d g e . Dirdeluate. Elute with -2 mL of nahswl. Collect 2 mL of cluuc into I grdualed - 15mLpolypropylenecmaifugetub0(Wvolume 2 d). Wyze -1- wingelectnuspy Lc/Ms/MS. 12.1 Meatbc smw anout of suh wdud, umple rad fodBed umplc into Ihc LCIMSIMS ryltaa. A d l n i i o a 8 t m d d must m e and follow all d W umpbr 12.2 Smuimnb of PPOA-OE to u IOUI Bve or mom consenention levels mudbeitlciud~din oa n d ~ ~ma .l 12.3 An c n h sd of ulibntionstandabmum be includedu Ibc beginning and at tho ond of umple KL Smuimnb must be i n t a r p d between every 5-10 m p l a . h o .homuivc. o ODtirS m of wlibruion Dududs may be iSi0d.a u tbc begbaing of a WI followed by calibration sundads iMonpaud way S-IO umpb (to for a tacond m of wldarde). In either cue, u l i b d m mutt be tho &rt rad *R injection in a umpk m. 12.4 Uw lbmr undrd cwc( for qumtimion. Lincu nnd.rd cutvea m paatcd for the mdytc by linourcprolionus& I/x waehting of peak a m v m w ulibruicn,danlmd EoIyQltRtio.wing M.uLjna 3.3 (or equivalent) WOlhrM ryltra 12.5 Sunplc rclponw lhould ao( a d s t x d d rUpOUWI. Any sunplu that cxotcdltadndr o r p m ~Wl d b e Iiuthadilutcdrad mnalyrcd. - -- Page 63 of 65 ___ -- Exygen Research Page 94 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Exygen Protocol Number: PO000760 hub Mnhod Nvmbn VWOIIU I I ANALYllCAL WTHOD Mahod of M y i r fortbe Lk&mhUhofP c r f l m m i c Acid (F'FOA) in Small Munmrl Sorum by Lc/Ms/Ms 13.1 13.2 13.3 13.4 13.5 13.6 Exygen Research Page 64 of 65 - . ._ Page 95 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Exygea Protocol Number: PoooO760 ' .ANALYTICAL .METROD Mabod of A d y d r brchsDorormirution of Pdvnooctraoic Acid (PPOA) in Small MvllmrlSaumbyLCMS/MS 14.3 Uw thc ppb. equationtocmval mC mountof PFOA found in np/mL to Exygen Research Page 65 of 6.5 Page 96 of 119 ? Interim Report #26 - Analysis of GroundwaterSamples Exygen Study No.: PO000760 E w e m R E S E A R C H 3058 ResearchDrive Phone: 814-272-1039 State College, PA 16801 Fax: 814-231-1580 PROTOCOL AMENDMENT Exygen Study Number; Amendment N u m b e r 1 Effective Date: 01/19/05 PO000760 Client Study Number: Page 1 of 1 DESCRIPTIONOF AMENDED SECTION 1) Analytical Procedure Summary V0001780:Section 9.1 2) Verification of Analytical Procedure None AMENDED TO 1) Add to Section 9.1: Section 9.1.6, Alternate weights of standards may be used to prepare alternate concentrations of stock solutions as necessary. Alternate levels of fortification solutions may also be prepared. 2) Low and high spiking levels of the analytes for each matrix may be altered depending on sample size available for extraction and/or to cover analyte concentrations expected in the samples. RATIONALE 1) Higherconcentrations of standards need to be preparedin order to spike the sample bottles at higher levels. 2) The sample size availablefor small mammal liver and serum was smaller than expected. Spiking at the predetermined levels in the protocol puts the spiked ' concentrationlower than the detection limit. Also, the analyte levels in the ground water samples are expected to greatly exceed the pre-determined spiking levels listed in the protocol. When the levels in the samples greatly exceed the spiking levels, an accurate recovery value cannot be calculated for the QC sample. Higher spiking levels in the bottles will cover the analyte concentrations expected in the water samples. IMPACT ON STUDY The LOQ is 100 nglg for a 0.1 g sample of small mammal liver and is 1000 ng/mL for a 0.01 mL sample of small mammal serum. Higher levels of spiking for the water samples will ensure that more QC recovery data can be used. ~~ - LIBRARY IO: WOOO1228-6 Exygen QAU Review ,kJ<I ADMINISTRATIVE FORM Exygen Research Page 97 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.:PO000760 ETBene R E S E A R C H 3058 Research Drive State College, PA 16801 Phone: 814-272-1039 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 DESCRIPTIONOF AMENDED SECTION Report. page 11 of 65 AMENDFD TO Insteadof one final report, interim reportswill be issued. m RATIONALE -,.- Due to the excessive sizes of the data sets, dient to receivedata in a timelier mwm. interim reports will 03,&kw be issued to allow the IMPACT ON STUDY The client will be able to receive and review the data more quickly. Study Director Management Signature ph.dpj=sj=- Exygen Manage e. LIBRARY IO: VOOO122E-E fG$@i 2 - Dale Date I;/,+$8-*-os Exygen QAU Review ,kls &/6 ~ ~~ ADMINISTRATIVE FORM Exygen Research Page 98 of 119 Interim Report #26 - Analysis of Groundwater Samples 08-02--2-0.0.5. - 0-0:llu .Cr_orK.STOW S.OL..UTlOWf Exygen Study No.: PO000760 w Exygen Research Page 99 of 119 InterimReport #26 -Analysis of Groundwater Samples Exygen Study No.: PO000760 w e n *. S~tRatsesCeoollergreh, OPAr1f6wWl phFoanxe::8114.227321--11053890 RESEARCH - Amendment Number. PROTOCOL AMENDMENT 49 m e 1dl fafecth Daw 07/18/06 Exygen Study Number P760 Client Study Number: NA Exygen Research Page 100 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygem Study No.:PO000760 ty@em RESEARCH 3058 Research Drive Phone: 814-272-1039 State College, PA 16801 Fax: 814-231-1580 Amendment Number: Effective Date: Exygen Study Number PROTOCOL AMENDMENT 5 09/13/05 P760 Client Study Number: Pagelofl NA DESCRIPTION OF AMENDED SECTION Analytical Procedure Summary: VOOOl780:'Method of Analysis for the Determinationof PerfluoroooctanoicAcid (PFOA) in Water by LC/MS/MS." AMENDED TO The reanalysis of two water samples, DFOS-GW-131R-0-040126 and DFB-GW-135L-0- 040121will be performed usingthe following procedure: An aliquot of the water sample is spiked with the surrogate standard andlor other matrix spike compounds as appropriate. Add methanol to the fortified aqueous sample. Mix and aliquot for LC/MS/MS 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=2mM ammonium acetate in water and B=Methanol (6 min equilibration). Use a 5 pL injection volume and a 0.3 mllmin flow rate. LC conditions may be further modified as appropriate for the system being used. An interim report will be issued that will include the exact details of the reanalysis. PATIONALE DFOS-GW-131R-O-040126 and DF8b-GW-135L-M)40121 could not be reported 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 negativeimpact on the study. More usable data may be obtained. Study Director Signature dlh Exygen ManagementSignature Sponsor Signature (if required) Date 4 Date Date Exygen QAU Review Oh. 9h3/0r ADMiNlSTRATlVE FORM Exygen Research Page 101 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen StudyNo.:PO000760 T0:*88142311588 NO.z!94 P.2 P:2'2 3058 b - r c h Drive Phone: Si4-272-1033 State College, PA 1W1 Fax: 614-231-1580 , Amydment Number: Effective Date: Exygen Study Number PROTOCOL AMENDMENT 5 OW1 3/05 P760 Client Study Number: Fmg.1061 NA ,DESCRIPTIONOFAMENDED SECTK)N Analytical Pmrdun Summary WOO178O~hodd Analysk fa the DobrminatianE PeAw-ctandc Add (PFOA) In Water by LCIMSMS,' AMENDED TO The reanalysis oftw water sampler, DFW-131RO.040128 and DFEGW-13614 oaOl21 will be perfamod usingthe fdlowing procedure: An diquat of the water sample is spiked with the sumgab standard andlor other maM spike compounds as appropriate. Add methanal to the farwkd aqueouo sample,. Mb and aliquotfw LUMSIMS anslysis, The LC mnditionsindud0the use Oia BeWlC78 2.1 x 50 mm cdumn, a gradknt bf 30% B to 100% B in 10 minutes, where b 2 m k ammonium acetate in water Md B=h4ethanol(6min equtlibrption). Use a 5 VLinjediw volume and 8 0.3 mUmh flow ram. LC andifions may be further rnodykd at appropriate for the system being osed. An interhn mpmt will ba issued th.t will indudc the madetailsof me reanalysis. RATIONALE DFOQ-GW-131RQQ10126 and DFSb-GW-G5L-M)40121 could net be npcwtcc No nwathre mpact on the me. MPACT ON Core usable SN beobteined. RECEIVED TIME 5EP.14. lW3fAM PRINT T I E 9 . 1 4 . 18:38AM Exygen Research Page 102 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.:PO000760 e . Amendment Number. Efkeive M a : E q p n Study Number PROTOCOL AMENDMENT P -01 dl Exygen Research Page 103 of 119 ~~ ~~ Interim Report #26 - Analysis of Groundwater Samples 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 PROTOCOL AMENOMENT 6 09120105 P760 Client Study Number: Pageldl 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 samoles for reanalvsis should be DFOS-GW-131R-0-040120 anc DF8b-GW-135R-0-040121. RATIONALE The correct samples are identifiedfor reanalysis. IMPACTON STUDY No negative impact on the study. StudyDiredorSignalyre, , Date m Mrector y ~ a g e m e n St ignature - Date Sponsor Signature (ifrequired) Dats Exygen QAU Review h . W 9/zo/os ~~~ LIBRARY I D VOOO1228-8 ADMINISTRATIVE FORM Exygen Research Page 104 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.:PO000760 T0:m6107817401 E8.388 P-. : 2'2 - P.2 Amendment Number: Elliecthm D a k ExVgen Study Number + PROTOCOLAMENDMENT ~ 7 8 0 UIOMMY ~mbet Page1 of1 NA The buo water JMlplrt for DFBbOW-135RSW012'1. TO be DFo9QW-131RQbQO120 and Exygen Research Page 105 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 - CHEH EHSR 236 16 SEP.2aEB85 2:- 651 733 1958 -5- 09/20 '05 1$:_4_6__Np.92002/02 , .- rD.JBB P.2 a RECEIVED TIM SfP.tB. 3:ElPfI P R I N T T I M .SEP.2B. 3:82pM Exygen Research Page 106 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 Exygen Research I Page 107 of 119 Interim Report #26 -Analysis of GroundwaterSamples Exygen Study No.: PO000760 CHEH EHSR 236 l B - 651 733 1958 JUL-V-CIIUJ W : l c rrMi-av uw. Lw W l ( ( 0 elf0 K w . a . m 4:- OClGplm 11/23 '05 1 4 : l l N0.979 tu:-% r s s Y a e -02/03 ~:5,4 --.-, NO.924 p.2 I RECEIVED T I E W.23. 332PM PRINTTIE Nov.23. 3:- Exygen Research Page 108 of 119 Interim Report #26 -Analysis of GroundwaterSamples Exygen Study No.: PO000760 EHS OPNS ENVIRONNENTAL LAB 1 .. .. .,. .0...6. 5.. 1. ..??.a 4226 ...I .......... . . . . ,.07./11/06 11:.:4.6.:,@.::0. 5/0.7. N.O.::68.1 . . .. .:, ... .. .......:... . . . . . . . . '..i ....... . . .. LIBRARY ID: wiab ' .. ., . . .. . .. . .' :..:.: .. . . . .....'.... . . .. ...... ... . ........ . . . . '. .. ;-.. . . I .. . ...,. .. ... .. . ./. .., ., . . . . ....... . . . ... ... . .... . . .. .. . . .. '.:.. . . . , . .. . .. . ........,..:.............: ..:.- .........+...................:....,-.. ..... . ................. *.,&..t..i.:..t::..'& ., . '-.* .; Exygen Research Page 109 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen StudyNo.:PO000760 \ RECEIVEOTIM JAN. a. 4 : m PRINTTIME J W . 8. 4:04PM Exygen Research Page 110 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.:PO000760 SP.20.- 1:- D(W;ENREm M.377 P.2 RESEARCH 3058 Research Ddve Phone: 814-272-1039 State Colkge, PA 16801 Fax: 814-211-1580 -?%tjeotsp-cDeviation Exygen plojsd#: ClientPmJeot# : DNlATfON FORM - Facnity Deviation P760P1131 NA P8ar1of1 w e of O c c m : 0316K)5 Deviation# : f if Ref-#: 05122 grvirtion (Indude Wt formothodJ end SOPS) Login#: NA Ccmtaiir # : NA Lot# : NA - - - - all son a d sedimenta m p k Dnrlyzed for pamnt 8olldn befae 07/07/05. a No blanks or duplleaft wgnt runaa mquindbysection @S. b. Some sample weightssrcrsd the allowable rang,01Ogl. Cauw: Pqamtion Anelysis lnsbumsnt Clht Rqurrt Other masbeenno negativeimp& onthe mdy. All of the permitaolld dues thatwere determined drrino me t h e m o d in question are cunsklered valM, dthoughthe SOPwae not fclbwd. In the wuly wised version of the SOP blanks and dupllcetm am no longer requimd. Atso, In the new SOP, Uw allowable amourd of sample to be used is c 20 g. All ofthe samples in quastion Inthis deviatbn weighed lesaman20 g. Theteohnldan anaiyzlngthe samples far p e w solids was fdlowlng the nw pmcedure bobre N was fopmdlyapproved. *e Actlom: A neW t!erskn $the SOP hM beenh u e d and appmwd (VO0004274). Exygen Research ADMINISTRATIVE FORM Page 1 1 1 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen StudyNo.:PO000760 N0.m P.3 3058 &%arch DdVe PhW: 814-272-1039 State College, PA 16801 Fax: 814-231-1580 Wula&ty D ~ i v p ~ : - OMP -X GLP byhst -- Ncfle 0.viatian Tvpo:(mcrude Mbrrnothodoand SOPS) -x Rotocal -Method -SOP w:NA Notebook reference: NA I / LIBRARY ID: MM)15so6 Exygen Research A D M I N I ~ l l V EFORM Page 112 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.:PO000760 N0.m P.4 3058 Research Drhre Phone: 814-272-1039 State College. PA 16801 Fax: 814-231-1580 DEVIATION FORM -Gxon#Prlm: jectspecifii~icltlon -FaclliiDeviation Exyeen PmjeCr #: P7601p1131 ~ldl Date of Occurrenee: 12n7/w Deviation#: 3/3. Client Project X : NA on Tuea: (/ndude V# livmslhads and SOPS) - - - Other NOM Olscriptlon: Pmtocol WI: 202-20 S & h 5.2.3 Notsbook lwlwmce: NA Login#: NA COrIklinerY : NA Lot#: NA ummawd * ! ~ k h l o n A l a n ineToluene) wa6 g k n the erpinbion data of WlJIoS,but RE544 (Toluene) used b makeSUI14 sxpind on 03/28/04. S K I14 IkaE usedtosilannlzed glasswireprepekdfor the tkh extmtion fmm 12/27/04through 01/07/05. - CIUSO: Preparetion -Anaiysio -Instrument -Client Request 0 t h ~ IDWi No IWWtIVe impadon the rtudy. TdumeWM u r d onlyas a sdwntfor the glasswarapreparation. Dimethyldlchlorosilane.w h i i ia tho coating agent, was not -ked. Cornutiw c\tifipnr; Dsviition issued. I LIBRARY ID:vwo16406 ADYNISTRATW FORM Exygen Research Page 113 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.:PO000760 3058ResearchDrive P h m : 814-272-1039 RESEARCH DEVIATION FORM - a!s!c& X Prcjed Specik D.vleUon - Facility Devistlon Exygen prollactw : P76O/P1131 meld2 Date of Ocarnence: 04)26106 D&tion#: 5 clienl Pmjed # : NA R&rmw#: 06476 m:(/ndudeMfor met)lods end SOPS) I - x Pmtml __ Method SOP W: NA - Summar d 0 . v l s t l o ~ The three sediment aamples inLE191(CO172892 C0172694) were ~fblnallyutrsdsd wine the sediment methodMoo1782. Poor rewwieawra obtainedfor PFOS, PFOAend "C PFOA Because of thk, Vu, ahaJyaponsarequeated the us% of an alternative extr#lbnfor theae m p a u d s , asMlowa: Dinct Inl.ction Mothod: Wore Ihe rampleawanwewedfor the extraclion,Uteyw(y0 mixed thoroughly byv4gorously hakhp the conlainor. Aone-gr~mportionof cedlment was weighed into lknill% centrifuge tube for the extradion. Ten mllilitsnof 1%acetk wid h melhanolwm adde4 to wch mpb.The rampleawan than shakenbyhand,vortexed. and sonhledfor themhutea. The umplei wen then mnlrifumfor -10 minuter nt -3aK)rpm. Each mpk was a w e d by LclMslMs eledrwpray. Usingthk mehod eCcapEabl0dnla waa obWned.forPFOS. but Me recoverleafor PFOA end "C PFOAwere Still poor. Another elternalivemethod waa lhmuaed for PFOAand `kPFOA, as follows: Altrrmtlvo 6PEMethod The .emplea Wl w w preparedh 1%wticsdd for me dhrctinjecllon me methodwere uced lor Wa exbadion. Fiw,milititeraof .achsamplewaa ahquoted into I5O-mL polypmpylennc e n f r y u ~tu~be and the volumowas taken to 40 mLwilh waler. The anmpkr were then c a n l r l w for -10 mhutea at -5M)o rpm. aupmmtent was lhmloaded onto a C,, SPE cartridge oondtknedwith 10 mLof melhanolend 5 mL of water. Theeluete waa discarded. Approximatelyfm rnYlilitenof m e l h a d was 8dded to the cam-. Five mlllom of duele waa cc4ected into a graduated 16 mL p o m y k n a untrlfugatub. Eachmpk was llnelyzed by LCIMSIMS .I-rPny. ADMINISTRATIVEFORM Exygen Research Page 114 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.:PO000760 Exygen Research Page 115 of 119 Interim Report #26 - Analysis of Groundwater Samples Exygen Study No.: PO000760 EHS OPNS ENVIRONMENTAL LAB .. &YC lf;Cm-io39. 8 6 5 1 778 4226 ..... . .07/11/06 . . . .. . . .. . ..._... ... .. . . . 11:46,f?.:0.4/0.7 N. O:.681 .... ., : : .j . . .. . .. . 3056 RWM& smtc Cotlpc, .PA i.beol . . . . '. ... .. .. : phohc: ..... F U : . .#.f.4.4; .3..1.rf.W. .' , , .. --' QMP GLP ' ' Oh* NOM ... - . .. . !Et%EiP .. . '. . ._ '. . . . ..... . . . . .. .. .. .. ..' .'. .. . .. . . .. . ,:.;. .. ..:. . ..:. .. .. . ..:.... . . .. .. . ..: :.. . . . ' . :.'.'. .. ..,:.. .. .....:. ... .. .. .. .. . . .. ... .. . . .. :.: ,: . . . .. . ... . . . . . .. ...:...;. ....I..:..;.... .;..... , .: .:.ii::..... :a.. :'.. ...... . ': . . . ' . .; : : . . Exygen Research Page 116 of 119 InterimReport #26 - Analysis of GroundwaterSamples Exygen StudyNo.:PO000760 EHS OPNS ENVIRONNENTAL LAB -. .. w.6651 778 4226 , fl 07/11 106 71 :46 :06/07 NO: 681 .. .. .. .. ..... .. .. .. .. . 3058 Itmead Drlvi' P m ' B l 4 ; State College, PA 18801 Fax: $...1, 4. ...' :.. ... ' .:.. . . . .. ,' ....a . .... . .... .. .. , : .. .. .. . ... .. ....... a,.. ' f d\k' 1... . . . ..... . . .................:..i..-..:....................... .. . ..L*I . . . . S.pol.m6.r+&.iyQ@O~ . . . . .:.. ..., s::D.,*.... .': , . . . . . Exygen Research Page 117 of 119 InterimReport #26 -Analysis of GroundwaterSamples Exygen Study No.: PO000760 . ; EHS OPNS ENVIRONHENTAL LAB . . .. ......~_......6......5. 7.17.8 4226 _, , ~ I . 3051w&hw~e State College,.PA. t6. 801 . . : .... . P .. . . . ......:........ . : . , .'., . . . . ...,:'.. -omurl.: . X".ProjoctSpecifkDevirtkn . ~. A&; .D. itr.al ........ ... F d l W. Owkuan ....... .: . . . .{..- .. ... ... ... . . . . ..... ... . ..: I;(,: . . ... .......... ............ .... . . . .... .. .. . . . . ...:.:... : :.:..:...:,....).......... . . . . . .. . . .: . .I I .. ..._-_._...._ ._ _ _ _ ._._. ip& . . .:. - . . . . . ..... .. ..D. m.b,. . :... ..2.. .. .. . .. . . . . . . . . . . . . .' . . ... .. .. . . . .I . . . . . . . . .... .. . . .:.., .... . . . . . . . ..... . .... . .. . . . .. . . .:.... ...,.._._- _ : : . . ' . i .,. ' .. . . '.. .. .. I .. ;,:.,;:? , , .- .....I .-.:.-z.:-i'.2. Exygen Research Page 118 of 119 Interim Report #26 - Analysis of Groundwater Samples Ol-08-200? 03:IBm FrorEfTMI SOlUTlfflS t Exygen Study No.: PO000760 MI P . O O ~ AmOa~ I I' RECEIVEDTIM JIW. 8. 4:82R1 PRIMTIME JW. 8 . 4 : e m Exygen Research Page 119 of 119