Document zQbRD7mdE0gNpkrdRG0nGwkJz

M & A 6 - 37*7 INTERIM REPORT #23 Analysis of Sediment Samples STUDY TITLE Analysis o f 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 DATA REQUIREMENTS EPA TSCA Good Laboratory Practice Standards 40 CFR 792 STUDY DIRECTOR Jaisimha Kesari P.E., DEE Weston Solutions, Inc. 1400 Weston Way West Chester, PA 19380 Phone:610-701-3761 INTERIM REPORT COMPLETION DATE May 2,2006 PERFORMING LABORATORY Exygen Research 3058 Research Drive State College, PA 16801 Phone: 814-272-1039 STUDY SPONSOR 3M Company 3M Building 0236-01-B-10 St. Paul, MN 55144 Phone: 651-733-6374 C: CO O to PROJECT . Protocol Number: P0000760 Exygen Study Number P0000760 Total Pages: 110 BEST COPY AVAILABLE OocumeniJ 000024 P-2 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0D00760 GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT Exygen Study Number P0000760, entitled "Analysis o f 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, with one exception. The laboratory control and control spikes for the direct injection and alternate SPE methods and the spiking procedures for the surrogate were not recorded promptly at the time of extraction. John Flaherty ' Principnal Investigator Exygen Research Jaisimha Kesari P.E., DEE Study Director Weston Solutions, Inc. Date Exygen Research Page 2 of 110 00002S Interim Report #23 - Analysis of Sediment Samples Jfci Exygen Study No,: F0000760 QUALITY ASSURANCE STATEMENT Exygen Research's Quality Assurance Unit reviewed Exygen Study Number P0000760, entitled, "Analysis of Perfluorooetanoic 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 phases1 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 . Final Raw Data Review and Interim Analytical Report Review Date Insnected 5/2/06 Date Reported to Date Reported to Principal Exygen Date Reported to Investiaator Management Studv Director 5/2/06 5/2/06 5/2/06 after Lead, Quality Assurance Unit s /toe / , Date lNote: 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 110 000026 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 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: ilm f k lj/ ; Flaherty Vice President Exygen Research s A /fiDate Exygen Research Facility Management: {I L , , ,__ Richard y<Mzzini President Exygen Research A) K tdurJ Date Study Director, Weston Solutions, Inc. gM tA /i/ JaisimhaTvesari P.E., DEE Weston Solutions, Inc. m Date Sponsor Representative, 3M Company: %2k Date Exygen Research Page 4 of 110 000027 ....... ...................................------------------------H-5. Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 STUDY IDENTIFICATION Analysis of Perfluorooctanoic Acid (PFOA) in Water, Soil, Sediment, Fish, Clams, Vegetation, Small Mammal Liver and Small Mammal Seram Using LC/MS/MS for the 3M Decatur Monitoring Program PROTOCOL NUMBER: P0000760 EXYGEN STUDY NUMBER: P0000760 TYPE OF STUDY: Residue SAMPLE MATRIX: Sediment TEST SUBSTANCE: Perfluorooctanoic acid (PFOA) SPONSOR: 3M Company 3M Building 0236-01-B-I0 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: 04/27/06 Interim Analytical Termination Date: 04/30/06 Interim Report Completion Date: 05/02/06 Exygen Research f ; f i Page5 of 110 000028 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 PROJECT PERSONNEL The Study Director for this project is Jaisimha Kesari at Weston Solutions, Inc. The following personnel from Exygen Research were associated with various phases of this interim portion of the study: Name John Flaherty Karen Risha Paul Connolly Christine Edwards Mark Ammerman Amy Sheehan Brittany Kravets Scott Crain Frances Crespi Devon Ingram Christa Gallant Title Vice President Scientist Technical Lead-LC/MS Technician Sample Custodian Associate Scientist Technician Technician Technician Technician Technician Exygen Research Page 6 of 110 000029 Interim Report #23 - Analysis of Sediment Samples SLl Exygen Study No.: P0000760 TABLE OF CONTENTS Page 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........... ............................................................................................. 12 4.0 ANALYTICAL TEST SAMPLES................................................................................12 5.0 REFERENCE MATERIAL.......................................................................................... 12 6.0 DESCRIPTION OF ANALYTICAL METHOD......................................................... 13 6.1 Extraction Procedure For Sediment............................................................................13 6.2 Direct Inject Extraction Procedure For Sediment...................................................... 14 6.3 Additional SPE Extraction Procedure For Sediment................................................. 14 6.4 Percent Solids Procedure For Sediment.....................................................................14 6.5 Preparation of Standards and Fortification Solutions............................. 14 6.6 Chromatography......................................................................................................... 16 6.7 Instrument Sensitivity................................................................................................. 16 6.8 Description of LC/MS/MS Instrument and Operating Conditions........................... 16 6.9 Quantitation and Example Calculation................................................................ 17 7.0 EXPERIMENTAL DESIGN........................................................................................ 19 8.0 RESULTS...................... 19 9.0 CONCLUSIONS........................................................................................................20 10.0 RETENTION OF DATA AND SAMPLES............ ................................................. 20 Exygen Research Page 7 o f 110 000030 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 LIST OF TABLES Table L Summary of PFOA in Sediment Samples................ Page 22 Table n. Summary of PFOA in Direct Injection Sediment Samples.............................23 Table III. Summary of PFOA in SPE Extracted Sediment Samples...............................24 Table IV. Matrix Spike Recovery of PFOA in Sediment Samples................................. 25 Table V. Matrix Spike Recovery of PFOA in Direct Injection Sediment Samples.......26 Table VI, Matrix Spike Recovery of PFOA in SPE Extracted Sediment Samples......... 27 Table VE. Surrogate Spike Recovery of 13C PFOA in Sediment Samples..................... 28 Table VE3. Surrogate Spike Recovery o f 13C PFOA in Direct Injection Sediment Samples.......................... ...... ....... ................................................................. 29 Table IX. Surrogate Spike Recovery o f ,3C PFOA in SPE Extracted Sediment Samples................................................................... ............. ...........................30 Table X. Total Percent Solids in Sediment Samples................................ .....................31 Exygen Research Page 8 of 110 000031 Interim Report #23-A nalysis of Sediment Samples Exygen Study No.: P0000760 Figure 1. LIST OF FIGURES Page Typical Calibration Curve for PFOA in Methanol......................................... 33 Figure 2. Extracted Standards o f PFOA in Methanol, 25 ng/L and 50 ng/L, Respectively.... ................................... .............................................................. 34 Figure 3. PFOA in Reagent Blank, 0.4 ng/mL Fortified Reagent Spk A, and 4.0 ng/mL Fortified Reagent Spk B, Respectively................................... 35 Figure 4. Chromatogram Representing a Sediment Sample Analyzed for PFOA (Exygen ID: C0172892, Data Set: 042806D).................................................36 Figure 5. Typical Calibration Curve for l3C PFOA in Methanol................................... 37 Figure 6. Extracted Standards of 13C PFOA in Methanol, 25 ng/L and 50 ng/L, Respectively............ ......................... ............................................................... 38 Figure 7. ,3C PFOA in Reagent Blank, 0.4 ng/mL Fortified Reagent Spk A, and 4.0 ng/mL Fortified Reagent Spk B, Respectively................................... 39 Figure 8. Chromatogram Representing a Sediment Sample Analyzed for l3C PFOA (Exygen ID: C0172892, Data Set: 042806D)...............................40 Exygen Research Page 9 o f 110 000032 Interim Report #23 - Analysis of Sediment Samples n. 10 Exygen Study No.: P0000760 LIST OF APPENDICES Appendix A Study Protocol F0OOO76O (Exygen Study No. P0000760) with Analytical Method V0001782: "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Sediment by LC/MS/MS", and Protocol Amendments.............................................41 Exygen Research Page 10 of 110 00.0033 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 1.0 SUMMARY Exygen Research extracted and analyzed sediment samples for the determination of perfluorooctanoic acid (PFOA) according to Exygen Method V0001782 (Appendix A). The limit of quantitation for PFOA in sediment was 2.0 ng/g. The standard sediment method (V0001782) was originally used to extract all of the samples. Use of this method yielded poor spiking recoveries for PFOA. Because o f this, a direct injection method (Section 6.2) was used, which also yielded poor recoveries. Finally, an alternative SPE column extraction (Section 6.3) was used to analyze for PFOA and ,3C PFOA. Analytical results for the analysis o f PFOA in sediment samples extracted according to the method based on dry weight are summarized in Table I. Analytical results for the analysis o f PFOA in sediment samples by direct injection, based on dry weight, are summarized in Table II. Analytical results for the analysis of PFOA in sediment samples by alternative SPE extraction, based on dry weight, are summarized in Table III. The overall average percent recovery standard deviation for PFOA in sediment samples based on wet weight was 73 16%. Fortification recoveries for the analysis o f PFOA in sediment samples are summarized in Table IV. Fortification recoveries for the analysis o f PFOA in direct injection sediment samples are summarized in Table V. Fortification recoveries for the analysis o f PFOA in SPE extracted sediment samples are summarized in Table VI. The average percent recovery standard deviation for 13C PFOA in sediment samples based on wet weight was 97 27%. Fortification recoveries for l3C PFOA in sediment samples based on wet weight are summarized in Table VII. Fortification recoveries for 13C PFOA in direct injection sediment samples based on wet weight are summarized in Table VIII. Fortification recoveries for 13C PFOA in SPE extracted sediment samples based on wet weight are summarized in Table IX. The total percent solids for sediment samples are detailed in Table X. 2.0 OBJECTIVE The objective of the analytical part o f this study was to determine levels of perfluorooctanoic acid (PFOA) in sediment according to Protocol P0000760 (Appendix A). Exygen Research tv . Page 11 of 110 000034 Interim Report #23 - Analysis o f Sediment Samples Exygen Study No.: P0000760 3.0 INTRODUCTION This report details the results o f the analysis for the determination o f PFOA in sediment using the analytical methods entitled, "V0001782: Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Sediment by LC/MS/MS." The study was initiated on November 05,2004, when the study director signed protocol number P0000760. The analytical start date for this interim report was April 27, 2006 and the analytical termination date for this interim report was April 30,2006. 4.0 ANALYTICAL TEST SAMPLES Three sediment samples (Exygen ID: C0172892-C0172894) were received on wet ice on April 22, 2006 from Tim Frinak at Weston Solutions, Inc. The samples were logged in by Exygen personnel and placed in refrigerated storage. Sample login and chain of custody information is located in the raw data package associated with this interim report. Storage records will be kept at Exygen Research. 5.0 REFERENCE MATERIAL The analytical standard, PFOA, was purchased from Oakwood Products, Inc. and was received at Exygen on March 17, 2005. The surrogate spiking standard, 13C labeled perfluorooctanoic acid (13C PFOA SP0004184), was received at Exygen on April 15, 2004 from the 3M Company. The available information for the reference materials is listed below. PFOA was stored ambient and I3C PFOA was stored frozen. Compound PFOA !3CPFOA Exveen Inventory No. SP0005444 SP0004184 Lot# 203 3507-195 Purity (%) Expiration Date 99 03/31/07 97 03/29/09 The molecular structure o f PFOA is given below: PFOA Chemical Name: Perfluorooctanoic acid Molecular Weight: 414 Transitions Monitored: 413 ->369 (for quantification) and 413 - 219 (for confirmation) Structure: Exygen Research Page 12 o f 110 000035 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 ,3C PFOA Chemical Name: 1,2-13C perfluorooctanoic acid Molecular Weight: 416 Transition Monitored: 415 --> 370 Structure: 6.0 DESCRIPTION OF ANALYTICAL METHOD The analytical method "V0001782: Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Sediment by LC/MS/MS" was used for the sediment samples in this study. 6.1 Extraction Procedure For Sediment Before the samples were weighed for the extraction, they were mixed thoroughly by vigorously shaking the container. A 5 gram portion o f sediment was weighed into a fifty milliliter centrifuge tube for the extraction. After fortification o f appropriate samples, 35 mL o f 1% acetic acid in water was added to the samples. The samples were vortexed and allowed to shake on a wrist action shaker for -6 0 minutes. The samples were centrifuged for ~20 minutes at -3000 rpm. The supernatant was then loaded onto a Ci8 SPE cartridge conditioned with 10 mL of methanol and 20 mL of water. The eluate was discarded. Twenty milliliters of methanol was added to the sediment samples left in the centrifuge tube. The samples were vortexed and allowed to shake on a wrist action shaker for another 30 minutes. The samples were centrifuged again for -20 minutes at -3000 rpm. The supernatant was then loaded onto the same Ci8 SPE cartridge. The eluate was collected into a 500 mL Nalgene Bottle. The column was washed with 4 mL of methanol. The wash was collected in the same bottle as the eluate. Approximately two Exygen Research * f Page 13 of 110 000036 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 hundred milliliters o f water was added to the bottles. The samples were mixed by shaking and loaded onto another Ci8 SPE cartridge conditioned with 10 mL of methanol and 20 mL of water. The eluate was discarded. Approximately five milliliters of methanol was added to the cartridge. Five milliliters o f eluate was collected into a graduated 15 mL polypropylene centrifuge tube. Each sample was analyzed by LC/MS/MS electrospray. 6.2 Direct Inject Extraction Procedure For Sediment Before the samples were weighed for the extraction, they were mixed thoroughly by vigorously shaking the container. A one-gram portion of sediment was weighed into a 15-milliliter centrifuge tube for the extraction. Ten milliliters o f 1% acetic acid in methanol was added to each sample. The samples were then shaken by hand, vortexed, and sonicated for thirty minutes. The samples were then centrifuged for -10 minutes at -3000 rpm. Each sample was analyzed by LC/MS/MS electrospray, 6 3 Additional SPE Extraction Procedure For Sediment The samples that were prepared in 1% acetic acid for the direct injection method were used for this extraction. Five milliliters o f each sample was aliquoted into a 50-mL polypropylene centrifuge tube and the volume was taken to 40 mL with water. The samples were then centrifuged for -1 0 minutes at -3000 rpm. The supernatant was then loaded onto a Cir 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 o f eluate was collected into a graduated 15 mL polypropylene centrifuge tube. Each sample was analyzed by LC/MS/MS electrospray. 6.4 Percent Solids Procedure For Sediment Percent solids were determined using the procedure indicated in Exygen method V0000427. Approximately 20 grams of sample was weighed into a pan. The weight of the sample plus the pan was recorded. The samples were then dried in an oven overnight at 104 2 C. Then the samples were transferred to a dessicator and allowed to cool for -15 minutes. Each sample was then weighed again, including the weight o f the pan. The percent solid for each sample was then calculated. See Table X for percent solids results. 6.5 Preparation of Standards and Fortification Solations A mixed stock standard solution of PFOA and 13C PFOA was prepared at a concentration of 1000 jig/mL by dissolving 100 mg of each of the standards (corrected for purity and Exygen Research Page 14 o f 110 000037 p. 15 Interim Report #23 - Analysis of Sediment Samples xygen Study No.: P0000760 salt content) in 100 mL o f methanol. From this solution, the following fortification standards were prepared: Cone, of Fort Stock or Fort. Volume Solution (mL) (pg/mL)1 1000 10 100 10 10 10 1.0 10 0.1 10 1o f PFOA and 13C PFOA Final Volume (mL) 100 100 100 100 100 Final Cone, of Fortification Std. (pg/mL) 100 10 1.0 0.1 0.01 A stock solution o f only 13C PFOA was also prepared at a concentration of 100 pg/mL by dissolving 10 mg the standard (corrected for purity and salt content) in 100 mL of methanol. From this solution, the following fortification standards were prepared: Cone, o f Stock or Fort. Solution (pg/mL)1 100 10 1.0 1o f 13C PFOA Fort Volume (mL) 10 10 10 Final Volume (mL) 100 100 100 Final Cone, of Fortification Std. (pg/mL) 10 1.0 0.1 A set of non-extracted calibration standards containing PFOA and l3C PFOA was prepared in methanol, as specified in Exygen method V0001782. The following concentrations were prepared Cone, of Fort Fort or Calibration Volume Solution (mL) (ng/mL)1 100 1.0 100 0.5 100 0.2 10 1.0 5.0 1.0 2.0 1.0 1o f PFOA and 13C PFOA Final Volume (mL) 10 10 10 10 10 10 Final Cone, o f Calibration Std. (ng/mL) 10 5.0 2.0 1.0 0.5 0.2 Exygen Research Page 15 of 110 000038 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 The standards at final concentrations of 10, 5.0,2.0,1.0, 0.5 and 0.2 ng/mL were used for calibration. Extracted calibration standards were prepared following method V0001780. The stock standard solution and all fortification and calibration standard solutions were stored in a refrigerator (4 2C) when not in use. Documentation of standard preparation is located in the raw data package associated with this interim report. 6.6 Chromatography Quantification of PFOA was accomplished by LC/MS/MS electrospray. The retention time o f PFOA was -11.1 min. Peaks above the LOD were not detected in any of the reagent blank samples corresponding to the analyte retention time. 6.7 Instrument Sensitivity The smallest standard amount injected during the chromatographic run had a concentration of 0.2 ng/mL of PFOA. 6.8 Description of LC/MS/MS Instrument and Operating Conditions Instrument: API 4000 Biomolecular Mass Analyzer Interface: Turbo Ion Spray Liquid Introduction Interface Computer: DELL OptiPlex GX400 Software: Windows NT, Analyst 1.4.1 HPLC: Hewlett Packard (HP) Series 1100 HP Quat Pump HP Vacuum Degasser HP Autosampler HP Column Oven HPLC Column: Thermo Fluophase RP, 50 rara x 2.1 mm Column Temp.: -30 C Injection Voi.: 15 pL Mobile Phase (A): 2 mM Ammonium Acetate in water Mobile Phase (B): Methanol Exygen Research Time fmin) 0.0 1.0 8.0 10.0 11.0 18.0 %A 65 65 25 25 65 65 %B 35 35 75 75 35 35 Page 16 of 110 000039 p-4X Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P000076Q 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 Monitored 413 -> 369 413 -> 219 415->370 Approximate Retention Time (min) ~11.1 min. -11.1 min. -11.1 min. 6.9 Quantitation and Example Calculation PFOA concentrations are quantitated in nanpgram o f PFOA per milliliter o f extract and then converted to nanogram per gram. The PFOA concentrations were quantitated in nanogram o f PFOA per liter for Set #3 (Set 042806D). Fifteen microliters o f sample or calibration standard were injected into the LC/MS/MS. The peak area was measured and the standard curve was generated (using 1/x fit weighted linear regression) by Analyst software using seven concentrations of standards. The concentration was determined from the equations below. Please note that the equations in this section are presented in a different order than the equations in the method (V0001782), but this has no effect on the final calculations. 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. Equation 1: Analyte found (ng/L) = fPeak area - intercept! x DF Slope Where: DF = Dilution Factor, factor by which the final volume was diluted, if necessary. For samples fortified with known amounts of PFOA and 13C PFOA prior to extraction, Equation 2 was used to convert the amount o f PFOA found in ng/L to ng/g (ppb). Equation 2: PFOA found (ppb) fPFOA found (ng/L) x SPE volume extracted! [sample aliquot volume x (sample weight / direct inject sample volume)] Exygen Research Page 17 of 110 000040 Interim Report #23 - Analysis o f Sediment Samples Exygen Study No.: P0000760 Equation 3 was used to calculate the percent recovery. Equation 3: Recovery (%) = . (analyte found (ng/g. wet weight) - analvte in control (ng/g, wet weight)') xl00% amount added (ng/g, wet weight) Note: For the PFOA recovery calculation, the "control" is the unspiked aliquot of the primary field sample. Equation 4 was then used to calculate the amount o f PFOA found in ppb based on dry weight. Equation 4: PFOA found (ppb) dry weight = PFOA found (ppb) x [100% / total solids(%)] An example of a calculation using an actual sample follows: Sediment sample Exygen ID C0172892 Spk D (Set: 042806D), fortified at 4 ng/mL with PFOA where: peak area = 236588 intercept 4630 slope dilution factor = 420 = 100 ng/g PFOA added (fort level) amt in corresponding sample (ng/g) SPE volume extracted (L) = =s = 4000 586 0.04 sample aliquot volume (mL) = 5 sample weight (g) =" 1 direct inject sample volume (mL) = 10 total solids (%) = 66.54 From equation 1: Analyte found (ng/L) From equation 3: PFOA found (ppb) = r236588 - 46301 x 100 420 ~ 55200 ng/L ~ (55200 ne/L x 0.04 L) [5 mL x (1 g / 10 mL)] = 4420 ppb From equation 3: % Recovery = (4420 ng/g - 586 ng/g) x 100% 4000 ng/g = 96% Exygen Research Page 18 of 110 000041 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 From equation 4: PFOA found (ppb) dry weight - 4416 ppb x (100% / 66.54%) = 6640 ppb NOTE: This value may be slightly different than that o f the 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 sediment samples in the laboratory after the samples were weighed for extraction. For sediment samples designated as laboratory matrix spikes, PFOA was also added at a known concentration to the samples in the laboratory after the samples were weighed for extraction. The sediment samples were extracted in one set. The set included one reagent blank and two reagent blanks fortified at known concentrations. The set contained three sediment samples. For each sample, two laboratory matrix spikes and a duplicate were extracted. 8.0 RESULTS Analytical results for the analysis o f PFOA in sediment samples based on dry weight are summarized in Table I. Analytical results for the analysis of PFOA in sediment samples by direct injection, based on dry weight, are summarized in Table II. Analytical results for the analysis o f PFOA in sediment samples by alternative SPE extraction, based on dry weight, are summarized in Table III. The overall average percent recovery standard deviation for PFOA in sediment samples based on w et weight was 73 16%. Fortification recoveries for the analysis of PFOA in sediment samples are summarized in Table IV. Fortification recoveries for the analysis o f PFOA in direct injection sediment samptes are summarized in Table V. Fortification recoveries for the analysis of PFOA in SPE extracted sediment samples are summarized in Table VI. The average percent recovery standard deviation for C PFOA m sediment samples based on wet weight was 97 27%. Fortification recoveries for 13C PFOA in sediment samples based on wet weight are summarized in Table VII. Fortification recoveries for 13C PFOA in direct injection sediment samples based on wet weight are summarized in Table VIII. Fortification recoveries for 13C PFOA in SPE extracted sediment samples based on wet weight are summarized in Table IX, The total percent solids for sediment samples are detailed in Table X. Exygen Research ' i i i Page 19 o f 110 000042 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 9.0 CONCLUSIONS The sediment samples were successfully extracted and analyzed for PFOA using an alternative SPE method, which yielded acceptable QC data. 10.0 RETENTION OF DATA AND SAMPLES All original paper data generated by Exygen Research that pertains to this interim report will be shipped to the study director. This does not include facility-specific raw data such as instrument or temperature logs. Exact copies of all raw data, as well as a signed copy of the interim analytical report and all original facility-specific raw data, will be retained in the Exygen Research archives for the period of time specified in EPA TSCA Good Laboratory Practice Standards 40 CFR 792. Exygen Research Page 20 of 110 000043 Interim Report #23 - Analysis o f Sediment Samples Exygen Study No.: P0000760 TABLES Exygen Research .fi't t M Page 21 of 110 000044 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Table I. Summary of PFOA in Sediment Samples Exygen ID C0172892 C0172892 Rap C0172883 C0172893 Rep C0172894 C0172894 Rep C lient Sam ple ID OAL-SO-BPPC1-0-QQOO D A L-S D -B P P 01-0 -0 0 0 0 * DAL-S D-BPP02-0-0000 D A L -S D -B P P 02 -0-0 000 * D A L -S D -B P P 03-<H )000 D A L -S D -B P P 03 -0-0 000 * A n alyte Found (ppb, n g/g) Dry W eight C 8 Acid PFOA P erfluorooctanoic Acid NR NR NR NR NR NR Assessed A c cu ra cy < * * NR NR NR NR NR NR ` Laboratory Duplicate NR = Not reported due to quality control result failures. See Table II for re-anatysis results. Exygen Research f Page 22 of 110 000045 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Table II. Summary of PFOA in Direct Injection Sediment Samples Exvasn ID C0172892 C0172892 Rep C0172893 C0172893 Rep C0172894 C0172894 Rep C lient Sam ple ID D A L -S D -B P P 0 1 -0 -0 0 0 0 D A L -S D -B P P 01-0 -0 0 0 0 * D A L -S D -B P P 0 2 -0 -0 0 0 0 D A L -S D -B P P 0 2 -0 -0 0 0 0 * D A L -S D -B P P 0 3 -0 -0 0 0 0 DAL-SD-B PP03-0-0000* A nalyte Found (ppb, ng/g) D ry W eight C8 A cid PFO A P erfluorooctanolc A cid NR NR NR NR NR NR Assessed A cc u ra c y [+ /-% ) NR NR NR NR NR NR "Laboratory Duplicate NR * Not reported due to quality control result failures. See Table III for re-analysis results. Exygen Research Page 23 of 110 000046 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Table III. Summary of PFOA in SPE Extracted Sediment Samples E x v a a n ID C0172892 C0172892 Rep C01728S3 C0172893 Rep C0172894 00172694 Rep Laboratory Duplicate C lien t S em ple ID A n a lyte Found {p p b , n g /g ) D ry W eig h t C 8 A cid PFO A P erflu o ro o c tan o lc A d d D A L-SD -B PP01-0-0000 QAL-SD-BPPOi-O-OOOO* D A L-SD -B PP02-0-0000 D A L-SD -B P P 02-0-0000' 881 945 927 823 D A L-SD -B PP03-0-0000 D A L-SD -B PP03-0-0000* 2390 2380 A tte s te d A ccuracy 30 30 30 30 30 30 Exygen Research Page 24 of 110 000047 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Table IV. Matrix Spike Recovery o f PFOA in Sediment Samples Sample Description DAL-SCFBPP01-0-0000 (C0172BM Spk D,400 ppb Spike) DAL-SD-BPP01-0-0000 (C0172892 Spk E, 4000 ppb Spike) DAL-SD-BPP02-0-0000 (C01S*S Spk G,400 ppb Spike) DAL-SD-BPP02-0-0000 (C0172M3 Spk H, 4000 ppb Spike) DAL-SD-BPP03-CM)000 (C0172S04spk j , 400 ppb Spike) D A L-SD -B PP03-0-0000 [C0172804 Spk K, 4000 ppb Spike) Amount Spiked fng/a) Amt Found in Sample <ng/g) wet weight C8 Add PFOA Amount Recovered fno/p) wet weight 400 4000 NR NR NR NR 40 4000 NR NR NR NR 400 4000 NR NR NR NR Recovery <%> NR NR NR NR NR NR Average: Standard Daviation: NR NR 'Sample residua exceeds Ihe spiking level significantly <3* spiking level); therefore, an accurate recovery value cannot be calculated. NR * Not reported due to quality control result failures. See Tatte V for re-analysis results. Note: Since this summary table shown rounded result*, recovery values may vary slightly from the values In the raw data. Exygen Research Page 25 of 110 000048 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Table V. Matrix Spike Recovery of PFOA in Direct Injection Sediment Samples Sara pie Descrtptlon DAL-SD-BPP01-0-0000 (com an spk d, too ppb spike) DAL-SD-BPP01 -0-0000 (00172192 8pk E,40000 ppb Spike) DAL-SD-BPP02-0-0000 (W17SS9S Spk G, 4000 ppb Spike) DAL-SD-B PP02-0-0000 (C0172SM Spk H.40000 ppb Spike) DAL-SD-BPP03-0-0000 (C0172BP4spk J, 4000 ppbSpike) D A L-SD -B PP03-0-0000 (C0172S94 Spk K, 40000 ppb 8 pite) Amount Spiked -W 4000 40000 4000 40000 4000 40000 Amt Found in Sample (ng/g) wet weight C8 Arid PFOA Amount Recovered (ng/g) wet weight NR NR NR NR NR NR NR NR NR NR , NR NR Recovery (% ) NR NR NR NR NR NR Average: Standard Deviation: NR NR ` Sample residue exceeds the spiking level significantly (3x spiking level); therefore, an accurate recovery value cannot be calculated. NR Not reported due to quality control result failures. See Table VI for re-analysis results. Note: SIrice th Is summary table shows rounded results, recovery values may vary slightly from the values In the raw data. Exygen Research Page 26 of 110 000049 Interim Report #23 - Analysis o f Sediment Samples Exygen Study No.: P0000760 Table VI. Matrix Spike Recovery of PFOA in SPE Extracted Sediment Samples Sample Description DAL-SD-BPP01-0-0000 (C0172SS2 Spk D,4000 nfl/g Spike) DAL-SD-BPP01-0-0000 (COI7WB2Spk E, 40000 n M Spike) DAL-SD-BFP02-0-0000 <CW?2*S Spk e , 4000 ng/g Spike) DAL-SD-BPP02-0-0000 (C0172003 Spk H.40000 np4oSpike) DAL-SD-BPP03-0-0000 |C0172094 Spk J, 4000 ne'0 Spike) DAL-SD-BPP03-0-0000 (C0172034 Spk K. 40000 ngfg Spike) Amount Spiked (nate) 4000 40000 4000 40000 4000 40000 Amt Found in Sample (ng/g) wet weigh 585 586 504 504 1060 1060 CB Acid PFQA Amount Recovered (no/o) wet weight 4420 25600 3860 25100 4140 23200 Average: Standard Deviation: Reoovery {%) 96 83 84 61 77 55 73 16 Note: Since this summary table shows rounded results, recovery values may vary slightly from the values In the raw data. Exygen Research r-j* i1 !>, , Page 27 of 110 000050 Interim Report #23 - Analysis o f Sediment Samples Exygen Study No.: P0000760 Table VII. Surrogate Spike Recovery of 13C PFOA in Sediment Samples Exygen ID Sem ple Description C0172892 C 0172892 Rep C0172892SpkC C0172892 Spk D C0172892 Spk E C0172893 C0172893 Rep C 0172893 Spk F C O t72893 Spk G CD172893 Spk H CO172894 C0172894 Rep C 0172894 Spk 1 C 0172894 Spk J C0172894 Spk K D A L -S D -B P P 0 1 -0 -0 0 0 0 D A L-SD -B PP01-0-0000 D A L -S D -B P P 0 1 -0 -0 0 0 0 DAL-SD-BPP01-0-DQOO D A L -S D -B P P 0 1 -0 -0 0 0 0 D A L-SD -B PP02-0-0000 D A L-S D -B PP02-0-0000 D A L-SD -B PP02-0-0000 D A L-SD -B PP02-0-0000 DAL-S D -B PP 02-0-0000 D A L-SD -B PP03-0-0000 DAL-S D -B PP 03-0-0000 DAL-S D -B PP 03-0-0000 DAL-S D -B PP 03-0-0000 D A L-SD -B PP03-0-0000 Amount Spiked (n g /p ) 4 4 4 400 4000 4 4 4 40,0' 4000 4 4 4 400 4000 ,!>c - p f o a A m ount R e c o v e red W e t W eight (ng/g> NR NR NR NR NR NR NR NR ' NR NR NR NR NR NR NR Recovery (% ) NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR Average: Standard Deviation: NR - Not reported due to quality control result failures. See Table VIII for re-analysis results. Note: Since this summary table shows rounded results, recovery values may vary slightly from the values in Ihe raw data. NR NR Exygen Research Page 28 of 110 00.0051 p. 29 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Table VIII. Surrogate Spike Recovery of 13C PFOA in Direct Injection Sediment Samples Sam ple Description . C0172892 C 0172892 Rep C O I72892 Spk C C0172892 Spk D C0172892 SpkE 00172893 C0172893 Rep C0172893 Spk F COI72893 Spk G C O I72893 Spk H C0172894 C 0172894 Rep C 0172894 Spk I C0172894 Spk J C0172894 Spk K D A L-S D -B PP01-O-OQOO D A L-SD -B PP01-0-0000 DAL-SD-BPP01 -0-0000 DAL-SD-BPP01 -0-0000 D A L-SD -B PP01-0-0000 D A L-S D -B PP02-0-0000 D A L-S D -B P P 02-0-0000 D A L-S D -B PP02-0-0000 D A L-S D -B PP02-0-0000 D A L-S D -B PP02-0-0000 D A L-S D -B PP03-0-0000 D A L-S D -B PP03-0-0000 D A L-S D -B PP03-0-0000 D A L-S D -B PP03-0-0000 D A L-SD -B PP03-0-0000 Amount Spiked (ng/g) 40 40 40 4000 40000 40 40 40 4000 40000 40 40 40 4000 40000 1sC -PFO A A m ount R eco vered W e t W eight _____ _________ NR NR NR NR NR NR NR NR NR NR NR NR . NR NR NR R eco very (% ) NR NR NR NR NR NR NR NR NR NR NR NR NR ' NR NR Average: Standard Deviation: NR = Not reported due to quality control result failures. See Table IX for re-analys)s results. Note: Slrtce this summary table shows rounded results, recovery values may vary slightly from the values In the raw data. NR NR Exygen Research . i* *'*.f . Page 29 of 110 000052 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Table IX. Surrogate Spike Recovery of 13C PFOA in SPE Extracted Sediment Samples Exygen ID COI72892 C0172892 Rep C0172892 SpkD C0172892 Spk E C0172893 C0172893 Rep C0172893 Spk C0172893 Spk H C0172894 C0172894 Rep C0172894 Spk J C0172894 Spk K Sample Description D At-S D-BPP01-O-ODOO DAL-SD-BPPOt-0-0000 OAL-SD-BPP01-0-0000 DAL-SD-BPP01-0-0000 DAL-SD-BPPQ2-0-0000 DAL-SD-8PP02-0-0000 DAL-SD-BPP02-0-0000 DAL-SD-BPP02-tK)000 OAL-SD-BPP03*<M)000 DAL-SD-BPP03-0-0000 DAL-SD-BPP03-0-0000 DAL-SD-BPP03-0-0000 Amount Spiked (ng/g) 40 40 4000 40000 40 40 4000 40000 40 40 4000 40000 13c -p f o a Amount Recovered Wet Weight (note) 52.0 51.0 3750 26800 47.8 51.8 3500 25200 45.2 40.3 3050 22100 Note: Sines this summary table shews rounded results, recovery values may vary slighUy from the values In Ihe raw date. Average Standard Deviation: Recovery (*) 130 128 94 65 120 129 88 63 113 101 77 55 97 27 Exygen Research Page 30 of 110 000053 p. 31 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Table X. Total Percent Solids in Sediment Samples Exygen ID CO172892 C0172893 CO172894 Sample Description DL-SD-BPP01-0-0000 DAL-SD-BPP02-0-0000 DAL-SD-BPP03-0-0000 Total Solids (%) 66.54 54.37 44.46 Exygen Research Page 31 of 110 000054 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 FIGURES Exygen Research Page 32 of 110 000055 Interim Report #23 - Analysis o f Sediment Samples Exygen Study No.: P0000760 Figure 1. Typical Calibration Curve for PFOA in Methanol 042 9 0 0 0 5 SPE.rdfc. (PFOA) " U n jr R giin p > * wnigM ing) y - 420 *-* 4.03**003 O'- o a s e s ) Area, counts Exygen Research Page 33 of 110 000056 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Figure 2. Extracted Standards of PFOA in Methanol, 25 ng/L and 50 ng/L, Respectively I X CM ZSM -f-PFOA (5taArd)4W.miS. mm* -tampt* 2 o f 4 t from M 2 M c n irfir A n*: fJZM coart* HUgMl 1.W*M3tpo KT; rt.tm h t 11.01 11D6 Exygen Research Page 34 of 110 000057 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P000076 Figure 3. PFOA in Reagent Blank, 0.4 ng/mL Fortified Reagent Spk A, and 4.0 ng/mL Fortified Reagent Spk B, Respectively I R MtnfaMtat cCtfoormtrtaiiS-)PFOA ( U n t m n ^ ^ n K K M m -tampli i fFt from HUXMfff EcU0>> I A m : 30296 unir Height 2 5 7 rK)03 op* RT: 11.0 min 11.00 Exygen Research . Page 35 of 110 000058 Interim Report #23 - Analysis o f Sediment Samples Exygen Study No.: P0000760 Figure 4. Chromatogram Representing a Sediment Sample Analyzed for PFOA (Exygen ID: C0172892, Data Set: 042806D) CM 722 -PFOA (itoJl*o\rr)-tllteiii.*amu-aampt* 19ef4*fromMZMEOMflr Ana:3S421ccuata HeJffkt: ZTtnOOScps R7t 11.0min 1103 Intensity, cp s Exygen Research Page 36 of 110 ooooSS Interim Report #23 - Analysis o f Sediment Samples Exygen Study No.: P0000760 Figure 5. Typical Calibration Curve for 13C PFOA in Methanol 04200608*4. 8PE.rib?13CPFOAfc"Unaar"R*0ruil6ftf l wtlghting);y40Bx+D.D010(f*OAOGO) Area, counts Exygen Research Page 37 o f 110 000060 Interim Report #23 --Analysis of Sediment Samples Exygen Study No.: P0000760 Figure 6. Extracted Standards of 13C PFOA in Methanol, 25 ng/L and 50 ng/L, Respectively I XCMMM+f 13CPFOA & sm f* * * )4 1 tW 7 ft*mu A n * : 1td01 c * * * t* H tigA ti ftffe+MQcp R 7 :11.$ m /* Z o f4 ttr o 1M2$MXw!Tr 11.02 11.00 Exygen Research Page 38 of 110 000061 interim Report #23 - Analysis o f Sediment Samples Exygen Study No.: P0000760 Figure 7. 13C PFOA in Reagent Blank, 0.4 ng/mL Fortified Reagent Spk A, and 4.0 ng/mL Fortified Reagent Spk B, Respectively I Hongont Control - 13CPFOA fimtnown) m .0/m .9nmn -*amph t o f4tfrom UQWKOMtff t* n t nottonne') (A 10.0-1 Tima, min I Raagarrl Spk A - 130 PFOA (00)415.0/370.0 im u u m pla 10 of 40 from 0428000 w tlf Ar*a:2729Q oounte H*ight2.C2+C3 cps RT: H U min 11-01 Tima, min I Raigafti Spk B - 13C PFOA(00)415.0/370.0 jm u * u m p l 11 of 48 from 042806&-*ff A ra i: 270001 oounfc Haight 22a+004 opr RT: 11.1 min o> a. 2j0a4 11.07 1.0*4 i 7 Tima, min Exygen Research <} Page 39 o f 110 000062 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Figure 8. Chromatogram Representing a Sediment Sample Analyzed for 13C PFOA (Exygen ID: C0172892, Data Set: 042806D) C*72*2 ttC P raA (tMt*OW*)415.V3Tt.e m u MKtfih 21o f Arm: 3149S7 coi/tts HUgJit; 2.9U*094tpt KT; 11.1milt from 042MtDMfT 11i>7 Intensity, cps Exygen Research Page 40 of 110 000063 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 APPENDIX A Study Protocol P0000760 (Exygen Study No. P0000760) with Analytical Method and Protocol Amendments Exygen Research ?, i Page 41 of 110 000064 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 ExygesProtocolNumber:P0000760 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: P0OO0760 Performing Laboratory: Exygen Research 3058 Research Drive State College, PA 16801 Phone: (814)272-1039 Sponsor Representative: Michael A. Santoro Director o f Regulatory A ffairs 3M Building 0236-01-B-10 St. Paul, MN 55144 Phone:(651)733-6374 P a g t ! c f 65 Exygen Research Page 42 of 110 000065 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Cxygen Protocol Number P0000760 DISTRIBUTION: 1) Jaisimha K m i, Study Director, 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 2 o jii Page 43 of 110 000066 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Exygen Protocol Numbei: P000076Q PROTOCOLAPPROVAL Study Tide: Analysis o f Perfluorooctanoic Acid (PFOA) in Water, Soil, Sediment, Fish. Clams, Vegetation, Small Mammal Livers and Small Mammal Serum Using LCAiS/MS for the 3M Decatur Monitoring Program Exygen Protocol Number: P0000760 APPROVALS Jaisimwi Kcsari, Study Director Weston Solutions Michael A. Si 3M Company Date a? -a x 'l D ata QualityAssurance Unit P u f s o fts Exygen Research Page 44 of 110 000067 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Exygen Protocol NumbT: P00CQ7W TABLE OF CONTENTS TITLE PAGE.................... ...... ...... ... ........................... DISTRIBUTION........... ......... ...... ..... ............ ............... PROTOCOL APPROVAL.......... .................................... . . ...................... 3 TABLE OF CONTENTS.................................. ..... ...................................... . ... ......... ........ 4 INTRODUCTION........................ ................ .................... .................................... ........................5 TEST MATERIAL.......... ................................. ................ .................... ...5 SURROGATE FIELD SPIKE COMPOUND........................... ....... ...I................. ...5 OBJECTIVE................................. ...................... ............. ......... ........... ...6 TESTING FACILITY..... ................................ ................ . ........ ............ ...6 STUDY DIRECTOR........ ................................... _............. SPONSOR REPRESENTATIVE................. ............................. PRINCIPAL INVESTIGATOR.... ............... ............................ PROPOSED EXPERIMENTAL START AND TERMINATION DATES........ ......................7 IDENTIFICATION AND JUSTIFICATION OF THE TEST SYSTEM....... ... ..................... 7 ^ SAMPLE PROCUREMENT, RECEIPT AND RETENTION...'...................... SAMPLE IDENTIFICATION....... .............................................................................. .ANALYTICAL PROCEDURE SUMMARY.... ........................................................ VERIFICATION OF ANALYTICAL PROCEDURE................................................. . .METHOD FOR CONTROL OF BIAS......... ..... .................................. !................. STATISTICAL METHODS....................... .................................... . GLP STATEMENT................... .............. ........................ REPORT....... ................... ...................... ................... SAFETY AND HEALTH.... ...................... ...................... ... _;.................. 12 AMENDMENTS TO PROTOCOL.____ ________ _____ .._ :... ............ .....13 DATA RECORD KEEPING.... ....... .. .................................. ............. ... .... 13 QUALITY ASSURANCE................. ......... .................. ...... .......................................14 RETENTION OF DATA AND ARCHIVING......... ........................................ ....................... ............ ...................... ......14 APPENDIXLANALYTICALMETHODS....... _ ............... .................................... ... 5......... .........................*..................... Pag* 4 o f 65 Exygen Research Page 45 o f 110 000068 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Exygen Protocol Nurobct: P0000760 INTRODUCTION The purpose of this study is to perform analysts 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 Program. The study w ill be audited for compliance with EPA TSCA Good Laboratory Practice Standards 40 CFR 792 by the Quality Assurance Unit of Exygen Research. TEST MATERIAL The test material is perfluorooctanoic acid (PFOA) end was purchased from Sigms-Aldrich. PFOA Chemical Name: Perfluorooctanoic add Molecular Weight: 414 Lot Number 23116HB Purity: 97.64% Transitions Monitored: 413 --> 369 (for quantification) and 413 ->-219 (for confirmation) Structure: F, F F 'OH SURROGATE FIELDSPIKE COMPOUND The surrogate field spiking compound is 1>C labeled perfluorooctanoic acid (1}C PFOA) andwas purchasedby 3M from Peririn-Ehner Life and Analytical Sciences. UC PFOA Chemical Name: 1,2-130 perfluorooctanoic add Molecular Weight: 416 Lot Number: 3507-195 Purity. 97% Transition Monitored: 4 1 5 - -3 7 0 P ag* S o f 65 Exygen Research Page 46 o f 110 000069 interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 ExygenProtocolNumber:P000C760 OBJECTIVE The purpose o f thi* study i t to perform analysis for perfluorooctanoic acid (PFOA) in water, soil, sediment, fish, clams, vegetation, small mammal livers and small mammal scrum for the 3M Decatur Monitoring Program using the current versions o fthe following Exygen analytical methods: VO0O178O: "Method o f Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS" V 0001781: "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) to Soil by LC/MS/MS" V0001782: 'Method o f Analysis Sot the Determination o f Perfluorooctanoic Acid (PFOA) in Sedimentby LC/MS/MS" V 0001783: "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS" V00017S4: " Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Vegetation by LC/MS/MS" V 0001785: "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Small Mammal Liver by LC/MS/MS" VOOOI786: "Method o f Analysis for tbe Determination o f Perfluorooctanoic Acid (PFOA) in Small Mammal Serum by LC/MS/MS" TESTING FACILITY Exygen Research 3058 Research Drive State College, PA 16801 Phone:(814)272-1039 STUDY DIRECTOR JaisimhaKesari P.E., DEE Weston Solutions, hie. 1400 W eiton W ay W est Chester, P A 19380 Phone: (610) 701-3761 Fax: (610) 701-7401 j.kesari@ w eBtonsolutions.com Paga 6 o f65 Exygen Research Page 47 o f 110 000070 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 fixygDQProtocolNumberPOO00760 SPONSORREPRESENTATIVE Michael A. Santoro 3M Company Director o fRegulatoryAffairs 3M Building 0236-01-B-10 St. Paul. MN 55144 Phone:(651)733-6374 PRINCIPALINVESTIGATOR John M. Flaherty Exygen Research 3058 Research Drive StateCollege, PA 16801 Phone:(814)2724039 jolm.flaherty@cxygcn.com PROPOSED EXPERIMENTAL START AND TERMINATION DATES It is proposed that the analytical portion o f this study he conducted from October 01, 2004 to December 31, 2005. The actual experimental start and termination dates w ill be included in the fatal rtjtort. IDENTIFICATION AND JUSTIFICATION OF THE TEST SYSTEM The following arethe test systems for tfaiastudy: Water (groundwater and surface water) Soil a Sediment Fish Clams Vegetation Small Mammal Liver Small Mammal Serum The samples w ill be collected by Weston Solutions. The control samples will be purchased and prepared by the testing facility. Purchase and processing Fegt 7v/6S Exygen Research Page 48 of 110 000071 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Eiygtn Protocol Number P0000760 details for the control samples w ill be included in the final report associated with this study. The test systems were chosen to access the environmental impact o f PFOA in the Decatur, Alabama area. SAMPLE PROCUREMENT, RECEIPTAND RETENTION Wales, soil, sediment, fish, clam, vegetation, small mammal liver and small mammal serum samples w ill be received at Exygen directly from Weiton Solutions. The detail* o fsample procurement for this study are outlined in the 3M work plan entitled " Phase 2 Work Plan for Sampling Environmental Media." The number and types o f samples collected w ill vary depending availability in the field. The total number o f samples received and analyzed for each matrix w ill be documented in the final report associated with this study. Water, soil, and sediment samples w ill be used as received without further processing at Exygen. These samples w ill be stored refrigerated at 2C-8C. Fish, clam, vegetation and small mammal liver samples w ill be processed according to the appropriate analytical method (fee Appendix I). These samples w ill be stored frozen at -UPC. Small mammal whole blood samples w ill be centrifuged in the field at the time o f collection and the serum fraction w ill be used for the study. Small mammal serum w ill be stored frozen at -HPC. The receipt and processing o f the samples w ill be documented in the final report and raw data associated with the study. SAMPLE IDENTIFICATION Prior to analysis, each sample w ill be assigned a laboratory sample reference number. The reference number w ill be unique and w ill distinguish each laboratory sample that is processed throughout the analytical procedure. Chromatographic data w ill be identified by the laboratory sample reference number. Sample storage conditions and locations w ill be documented throughout the study. Page 8 c f 65 Exygen Research 'S. t** !1 f ' Page 49 of 110 000072 Interim Report #23 - Analysis of Sediment Samples -sa Exygen Study No.: P0000760 ExygenProtocolNumberP0000760 ANALYTICALPROCEDURE SUMMARY Reference*: V000178Q: "Method o f Analysis for the Determination of Perfluorooctanoic Add (PFOA) in Water by LC/MS/MS" V00017B1: " Method o f Analysis for the Determination o f Perfluorooctanoic Add (PFOA) in Soil by LC/MS/MS" VOOOt782: "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA)in Sediment by LC/MS/MS" V0001783: "Method o f Analysis for the Determination o f Perfluorooctanoic Add (PFOA) in Fish and Clams by LC/MS/MS" V0001734: " Method o f Analysis for the Determination o f Perfluorooctanoic > Acid (PFOA) in Vegetation by LC/MS/MS" VOOOl73$: "Method o f Analysis for the Determination o f Perfluorooctanoic . Acid (PFOA) in Small Mammal Liver by LC/MS/MS" VOOOl786: "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Small Mammal Serum by LC/MS/MS" The above methods use analytical conditions capable o f separating the isomers o f PFOA. The final report w ill include the isomers summed into total PFOA found. VERIFICATIONOFANALYTICALPROCEDURE A laboratory control sample w ill be used for the preparation of fortified control samples. The test substance w ill be made into solutions as per the method, and added to the matrices via a miciopipettc. For water sampling, Exygen w ill supply one bottle persample collected. The bottles w ill be 500 mL precleaned Sci/Spec Premier wide mouth HDPE bottles. These bottles have been routinely used for PFOA sample collection at the testing facility and have been shown to be free o f PFOA. A ll containers used for water sample collection w ill be shipped to the sample location containing 160 ng o f 15C PFOA (equivalent to 500 ng/L in the final sample). Samples will be added to each container to a volumetric fill line at 200 m L A field duplicate, a low field spike and a high field spike o f each sample w ill be collected. The low and high field spikes w ill contain PFOA as well as perfluorobiitanesulfbnate (PFSS), perfluorohexanestitfonate (PFHS), and perfhioicoctanesulfonatc (PFOS). These compounds are included in the solutions used to spike the samples. The results for PFBS, PFHS and PFOS w ill not bereported in this study. Exygen w ill supply one field blank (control water) and two field blank spikes (control water fortified wilh PFOA at a low Page 9 q f6 J Exygen Research Page 50 of 110 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 ExygftnProtocolNumber:P0760 and high level) for every twenty sample collected. At the testing facility, each water sample (excluding field duplicates and field spikes) w ill be extracted in duplicate and w ill also be fortified at a low and high concentration with PFOA and processed through the described procedure to determinemethod accuracy and to check for bias. For soil, sediment, clems, and vegetation, Exygen w ill supply one 500 mL precleanedSci/Spcc Premier wide mouth HDFE bottle per sample collected or a zip-seal bag. A ll containers/bags used for sample collection w ill be shipped to the sample location. Samples w ill be added to each container or bag in the field.' A t th testing facility, each sample w ill be extracted in duplicate and w ill also be fortified at a known concentration with PFOA at both a low and high level and procesaed through the described procedure to determine method accuracy and to cheek for bias. u C-PFOA w ill also be added to each sample in the laboratory prior to extraction at the level specified below. ' For small mammal liver, Exygen win supply a $0 mL polypropylene contrifoge tube. Forsmall mammal serum, Exygen w ill supply acollection kit for each sample contaming serum separator tubes (red top), vacutainers. needle holders and needles, transfer pipettes, and polypropylene tubes. At the testing facility, each Hverand serum sample w ill he extracted in duplicate and w ill also be fortified at a known concentration with PFOA at both a low and high level id processed through the described procedure to determine method accuracy and to check for bias. n C-PFOA w ill also be added to each sample in the laboratory prior to extraction at the level specified below. Low and high spiking levels for each matrix aredefined below: M atrix Low PFOA High PFOA "C-PFOA Sulking Level Spiking Level Spiking Level Water SOOng/L 5000 nsn . 500 n tfL Soil 4 ngte 40ng/g 40ng/g Sediment 4 nate 40 ngte 40 ng/g Fish 10 nate lOOngte lOOng/g Clams 10 nate lOOnste lOOngte Vegetation 10 nute lOOng/g lOOngte Small Mammal Liver lOng/g lOOng/g lOOngte Small M ammaliSerum lng/g 100 ng/mL 100 ng/mL Recoveries am anticipated to -b e between 7CW4 an d 130% o f the fortified levels; however, the exact precision and accuracy w ill be determined by the analysis o f the quality control samples described above. A statement of accuracy w ill be included in the final report. Prior to sample analysis, a laboratory control spike study w ill be conducted in water to justify the use o f l3C PFOA results to establish precision and accuracy o f PFOA samples. The following samplesare Id be prepared: Page t0 oj 6S Exygen Research Page 51 of 110 000074 Interim Report #23 - Analysis o f Sediment Samples Exygen Study No.: P0000760 Exygen Protocol Number P0000760 Two control water blanks Two control water samples fortified at 250 ng/L with 1JC PFOA Two control watercamples fortified at 250 ng/L with !1C PFOA and at 250ng/L with PFOA. Two control water samples fortified at 5000 ng/L with ,3C PFOA and at 5000 ng/L with PFOA. The above samples are to be extracted and analyzed according to method Y0001780 entitled " Method o f Analysis for the Determination of Perfluorooctanoic Acid (PFQA) in Water by LGMS/MS." METHODFOR CONTROL OF BIAS Control o f bias w ill be addressedby taking representative sob-samples from a homogeneous mixture of each matrix from untreated control samples, and by analyzing at least two levels o f fortifications. STATISTICAL METHODS Statistics w ill be limited to (hose specified in the subject methods and to the calculation of averagerecoveries, a* applicable. GLP STATEMENT A ll aspects o f 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 w ill outline any deviations in the study from those standards. This statement w ill be signed by the Study Director and Sponsor Representative. REPORT A final report w ill be prepared by the principal investigator or their designee at the conclusion of the itudy. The report w ill include, but w ill not be limited to, the following: Page I I o f f! Exygen Research Page 52 o f 110 OOOOV Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 . The name and address of the Study Director, Sponsor Representative, and o fdie testing facility. A statement o f GLP compliance {any related documentation, such as chain-of-custodyrecords, most he in the study records). .. The signed and dated statement by the Exygen Research Quality Assurance Unit regarding dates o f study inspections and dates Sndings were reposted to the Study Director and Management. A description o f the exact analytical conditions employed in the study. If the subject method was followed exactly, it is necessary to include only a copy o f the analytical method. Any modifications to tins method w ill be incorporated into the report. I f the method is photo-reduced, the project number and page number mustbe included on each page. Description o f the instrumentation used andoperatingconditions. e A ll results from all sets analyzed. Control and fortified samples w ill be identified and the data table w ill include sample number and fortification level. Representative chromatograms for each analyte in each matrix, including chromatograms o fa standard and acontrol sample, and a chromatogram at a fortification level. The location o f the analyte peaks w ilt be clearly identified in all chromatograms. A ll circumstances that may have affected the quality or integrity o f the data w ill be documented in the report. Locations where raw data and the final report are to bo archived. Additions or corrections to the final report shall he in the form of an amendment signed by the Study Director. The amendment shall clearly identify that part o f the report that is being altered and the reasons for the alterotions. The amendment w ill be signed and dated by the Study Director andthe SponsorRepresentative. SAFETY AND HEALTH Laboratory personnel w ill practice goad sanitation and health habits. Every reasonable precaution shall be taken to prevent inadvertent exposure of personnel and the environment to the test or reference substancefs). Page i2 q f( i Exygen Research Page 53 of 110 #076 Interim Report #23 - Analysis of Sediment Samples f*"64 Exygen Study No.; P0000760 Exygen Protocol 'Number: P00007&0 /-* ameendments to protocol A il significant change* to the analytical protocol outlined here w ill be expressed in writing, signed and dated by the Study Director and Sponsor Representative. Amendments usually w ill be issuedprior to initiation o f study plan change. However, when a change is required without sufficient time for the issue o f 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 aa soon aa possible. In this case, the effective date o f the written amendment w ill be foe dale o f the documented change. Copies o fthe signed amendments w ill be appended to ail distributed study plan copies, the original amendment w ill be maintained with the original studyplan. Any deviations from the study plan or from the analytical method asprovided w ill be documented and reported promptly to foe Sponsor Representative. DATARECORD KEEPING Records to bemaintained include the following (as appropriate): Sample tracking sheet(s) Sample receipt records, storage history, and chains o fcustody History andpreparation o fstandards (stock, fortification, calibration) Description o fany modifications to the method Instrument run sheets, bench-sheets or logs f Analytical datatables A ll chromatographic and instrumental conditions Sample extraction and analysis dates A complete listing o f study personnel, signatures and initials Chronological presentation of all study correspondence Any otherdocumentation necessary for the reconstruction o f the study C hrom atogram s- A il chromatograms w ill contain the following: Sample identification, injection date, arrow or other indication o f the area of interest, and injectionnumber corresponding to the run. Additionally, fortifications w ill include foe amount o f analyte added and the sample number o ffoe samplethat was fortified. Analytical standard chromatograms w ill additionally include foe concentration (e.g., ggfaiL). Page 13 o f 63 Exygen Research Page 54 of 110 000077 Interim Report #23 - Analysis of Sediment Samples p. 55 Exygen Study No.: P0000760 ExygenProtocolNumber:P000760 As part o f the documentation the following sheets w ill be included in each analytical set; a run sheet liltin g the samples to be run in the set. and an instrument conditions sheet describing the instrument type and operating conditions. A ll applicable requirements for reporting o f study results as per 40 CFR 792.185. QUALITY ASSURANCE The QA Unit o f Exygen Research w ill inspect the study at intervals adequate to asaure compliance with GLP's, and w ill report the findings o f audits to the Study Director, Exygen Management, andthe Sponsor Representative. RETENTION OFDATAANDARCHIVING A ll hard copy raw data, including, but not limited to, the original chromatograma, worksheets, correspondence, and results shall be included with the data package submitted to the StudyDirector. These w ill be archived with the original study plan, amendments, final report, and all pertinent information from the Sponsor. The testing facility shall keep all electronic raw data and any instrument, equipment, and storage logs for the period o f time specified in 40 CFR 792.195. An exact copy o f the materials submitted to the study director w ill also be kept at Exygen Research. Exygen w ill obtain permission from the study director before discarding or returning samples. Exygen Research Arg Ho/6} Page 55 of 110 000078 Interim Report #23 - Analysis o f Sediment Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 APPENDIX I ANALYTICAL METHODS VOOOl780; `'Method o f Analysi* for the Determination o f Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS" V0001781: " Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS" V0001782: "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Sediment by LC7MS/MS" V000178J: "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Fish andClams by LC/MS/MS" V0001784: "Method of Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Vegetationby LC/MS/MS" VQO01785: " Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Small Mammal Liver by LC/MS/MS" V0001786: "Method o f Analysis for the Deicmunation of Perfluorooctanoic Acid (PFOA) in Small Mammal Serum by LC/MS/MS" Exygen Research Pag* 15 o f 65 Page 56 of 110 000079 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Exygen Protocol Number: FOOOQ760 AMNfAtWLNYwTInCbAenL MVOEWTH17O*0D M4fcp4tAnaJytiittrthDaftrmlafcaytLkOaaMfSP/wMflMfl wctwiQieAdd(TTOA}lnWater AnalyticalTatis]Facility: EIQiSya*aRieltseeaaerMchcDbnvc State Ctafe, PA lOI ApprovedBy fTM^aidlCnauaaolLU*y4tcr.LJCMLS,.B'xygtttomith Date /WJUohFnFltiULMboty ' VViiec*PFrte*m*idemOparMta*. btyg Roetefcb A . Exygen Research TotalPafaa: ? PftgC 16 Bf65 Page 57 of 110 000080 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P000760 ExygenProtocolNumberPODD0760 M**4H*wVO06ino [ /W AlYnCAl MKTBQft Me*edfAailjmferAeDmnmatfLuOBWcfSP/MerQSucirooclafiolcAddPFOA)inWaterby 1.0 Scope HaSdpidocatbronynteeHOdi|xhieDIPeetetrefcotrommr(apLntCoce/yM*LSifqeMnrSifde)eCDMtovaeeMmttroa.t*otHraopdfc^?uecsotiqteptlieodaotofp*nfltimNvdenonctMtMic 3.0 Safety 2.1 Alwayvobavvelifelaboratorypractice*. 12 CmtfoeapportiateMSDSbeforehaadliafanychemicalfarpropersafety 3.0 SmeblUfiiinMt 33..11 ATinifelaaieigi4d0ajcmaLaoiwflaiinifaIrunepetiortefodrfcneitmrveeinttam.iiiiiIm cempentace.J J Ssnplce atomd xe&ifcnted ahrok! te allqwed \q equilibraie so roarn 33..34 nAAalliayMt-Mlaaipainlepd*lqnfefiNfeffnlltloKpelnlhntoautfialMpamtia|tiettfyeedemfeixbrewtdhieMtefbxecmfftoeectiiiontgntru.ifamjepdleadi-f3o0rc0x0trrapmctiofbiir-3 3.4 pSraqnjepctLecoOectionprocedimiwillterprctodiotheamplinpplanfar thit 4.0 fteefcntem i Simfeed 4sl A4.23 MAWxErathDtooorni-ouKlm-PHALFCcLeiCntadftne*d-Ac.C.3.Rei^aMGrade A4 PnflBurWttteioicAcid-Slpna-Ajdrich 5.0 TiuhumfiiimdElJMipmOli 3.1 Abi|hpertbnnanct liqnd doroauuojrapbcapabl*of pumpugcpto2 pioLItcooauetotiojjpdyfoodatiartnddemvMiraiiMatSapvaectvtoomeMvp*o(LcOtorMcaSpAabiSh)t.ofiojectins3*200 3333133......234378 2D31AA!302rietduonn3eaLeL-vlt*myttededttMa*Llit*ritnptUpobHeaoVtcodPnirrUaoLlttflnaCofoe^ciprvpPlofcrielaoUwqllw]jyp(Ii*5pidrffaoer0eo.p-bip!eyiIiOyhUlffeebrOnneropuaeLecda}lew*fkea1f.ri0|tatriof0iftoee-20j^fae0.r000ttoout0iobbL0aeo)1.a*..%u.3 qwmituion. 331..19110 ADWuiqatowrmpeIpeStobcMlippi>^(M1*0VH0ea*e1c.0600e|i(L)anIdC1H0*1m00eptLoXtriwdtpibed.ippoeahfetip*. H vi*n Pag* i 7 o f t t Exygen Research Page 58 o f 110 000061 Interim Report #23 - Analysis o f Sediment Samples Exygen Study No.: P0000760 Exygen Protocol Number: P000076 MMMUmtm VQOQ1790 AlVLYTICALMrniQP McodtfAttlpiftirtgeDiMmlnaoBofNrfluonoeianolAdd(PPA>inWsmtby 59..112) SCePEeVi^KUtUeBtfnduoliiJtbflkyLiaB|WLfOtyprapyten*tabeaatWOOipuv 6.0 ChrowloyoplieSylt 6.1 66..2) {MTMGAPenemWooatMitl:yiyaatbiP2rcmPta5fPah0lerCa5a:M-^oi0olvXu5(fABbm2Tl)))*C3:n:0:F}li2MwnepktfhcMaaniAeeoB!iPno(KoiayonAac*cSuctoieUntWifica),ts2r.1nmx50mm,5p 0.D IJ> 22U30>05 26662L35555A &33377355$5 Ffmlolw0A.3Rninitet 0000...3333 66..76 lQojouicnttlooaotiVeotf:wnPn:klA5rpeL(-cnarotcbnetUiltcturuuilinltkooiiliBbnrsticohnMcu5rv0ep.L). 6.6 JfcacTUw: -23miiiie. qTpbdmeJbsoevetboHn7d1itfCc*t#y*rrtwmi.tndad guJtkandmybecimjedinenterto 7.0 m m S ym 7.1 Mods:!Dc|iiny>feptiv*MRMmo4t>iMmitarii44])--069nv'*. The tk v t coodtlioni m loaded u a guide n d may be d m ged in order i (f tim u e th i MSMS ykm . 1.0 PnjNuaiioaofSdlirtiMi 1.1 MctnlPhiM UM v2nuM*davonwMoetaimtelaocelOtaOteOamwLaotfewrDuerp.reparedbyaddin0.154*of A lternate vqNiim * m ay be prepared. Exygen Research ' * * f *>? M j Fgt 18 (ifto Page 59 of 110 000082 Interim Report #23 - Analysis o f Sediment Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 gyywlu -ili hirfiwV0Q0I7W [ Sfrfafcd|ofAnajyiafarftDeAfanNuAisLaYtLicTGqICMoAfP&LeMMrfSluoEroocTluHoicOAdDd(P_FO_A_)i_nW__e_rby] 9.1 SlandsrdStoefc/FeriUteationSolution 911 orfrolernihfatunlojlaklfawaelssetUkanoi-edmlaudhLo(LeftDooPnf-eB1cb0itdo0tffomerfpouLritoyr)PeFnOddAtobtyewtoeig10h0inngiL10wmithg 9.1.2 miAelLOo1p2f5gfe/mneloOl*LQLieDpnPgiBfictobsLotiettsleno,ltuotifotnnfalonoitfnppiiOl vAoliupmreepocfre10d0bwyWithiongrtinagno10i 9.1 J A1.0pgfaLtonificationsolutionofPPOAisprepvedbybrtrv|1ngnr 9.1.4 m1A10L2.15oapffgLt*faL1LD0Ppfogrtbfifaioclt.atlertoio.tantiroontattoionfoiufPlPvOoluAmieporefp1a0re0dwbityhbnrwinighianrgallmu fAa1L2e5fnfaLeLlD.0PBboldss.olutiontoeflmlvolumeof100withtneihuolin 9.1.5 wA10OfamiOwLliploignfAtaOnL12t5o0.co1tiiLfijeigLaDttaioPLoBSsbDoot&tatltoeist.tfalitotfoPF4OfiAnt1ivporslupmitresdobfy1b0r0inwjieithg 9.1.0 efaTprbmprewthademiodMctkfeeBlyei4df*pfCereretyinfilsdolatiuffieeen.stkaoblheiHfinMieninsixoubnfuttioofpeedriiondAonfft6gmeroAnithtrsti 9.2 SuedsrdCAUfeatieaSplutiees 92.1 LeidQeonlMfttcio1sdMtittoStsimteanitpdibalanrdif*ltoticlupnrodcetfsdstedArthtrpoeupgshesthdeintrtiaFcUtiConwpartoecr.eduTrhee, 902 Tprhsepsfaroldowisinngeeidsesd.lypdenlQuonplecadtttiosialeowentnaioatmsybe *oTcXCSoJhCoPMbelunoceti111tomneJe11I6Q000dn00Qzt00rnwfstacrbMeaeroiebdikotd)ei*,cddocFuaaboVltierob(434o1l21#nin00M000ktltLuhr000t0asw)etdeluaConofKlno_aPefstea_ShdVnreatiitrefndliedcut444.444iwdualt000e000mlaCi_bfasobroa_Lnfftft)itao_onl sstCaSunetauCdecaaituienrlP1b5i2edLb250e05im00ri500ddn0a1000fumttaiosrlxiqnonuPngaodtfhteoXXXXXXXafSxOOcQCCCCCiettawamamwiattztiaosomlmneiwnbmmttdianddknndHddrndddnddiyytdkdwyimrlyyaD<yr)yj..HyAS1J2l^iPE). Page t9 o f 65 Exygen Research Page 60 of 110 000083 Interim Report #23 - Analysis o f Sediment Samples Exygen Study No.: P0000760 Exygsa Protocol Numhtr P00007W WittedNwadbVO*0IHO ANALYTICAL MFTIfQP M*ttw6ofMtlynifortfIfotsraaticaofFerikttmtiSBkAdd(PFG)bWuby LC/MSMS 9Z1 ^acMtafmMaBddit>drifiwotdentxmte(dn.aiub(c*)SBiHiJuL8[Bii9slwUhBch ' 914 iS(2te*f*ClUlfetx{,ttCe|tVedpcl9atWlQ9mWiDNBkas.tandardsinl^mLyetypopylcnttube* 9.2.3 AhmatevctonmindMfteeiOrMtaaofilaiadaidsmaybeprepared*a nwted* 19.0 fto ca S g tV f 10.1 Ifo2 RpoEienaoaemfocaoqchrrsTniabqtfAasSaumttKsatfbMolfistctyoetittiffMoaUfouvdirwunlwSyrparlofetiolnd(rkernpntmeioolnaxdbwtnrbealnafadcobttceoroMhdtarh.anni(tsooifjpcrapyfncrudetUmsusiJpnieoylgitMca2Wt0e(lsaLobraCmImctwoipan)ittkecreorm)al iv(twpfioniiklcleltubw)edoeteoprietcvci(ticrfciiomluVdt 11.0 ganviteErinflfai . U.l itaHeianww5w0iUaiXs*L0fomtyi.porofpytaiaoeranptroifortfieontuobfeesw(nfipxteitydiluateodeemdeodm,reLplawciethfidwaenrd 11.2 11.3 lCfoeOaondowdsleaiodmobpydle5omnCcl*.oonSfdFBitEiPonLusCrdrlwdCagtseSsrT(~B(12c&adrrtor6ipdmgfreeLc.>)f.btaDypcpmiofltlotinktegtacts1o.0lummnl.rmunetdhraynol 11.4 E15luntetipwoklybp-r5epay*Ltaa10o0a%amaifcOfcimI. C(finolaelcvts5homnaLo5femluLa>teintograduated 1LJ AnalyseimptesucogtiectroaprtyL0MS/MS. 12j0 Cbsoaatopnpby 12.) ifqjMae&tlhM8tamsyesatemmo.unAtocfaefaibcmhstjatanndsatradn,dsaarmdpmleaaxndpfroerctiefideedasanmdpfloeliomwotahell 12.2 125 analysed ucDpUa- AbmUSnxantuwMwpaiaanaAtbtatnarf*rnbdlaianeifoccdfalaapPnebdKPdteafQtcowdanAeftktetMfeaonMxieBtrkiaiefuteWecanwotredyd^fdtiwitoMekdcisfialaO^Jamlafmosacac*mtmm-yitiotppbl*leeet--sr.vtaerqcAtaet.dBscatewarEddntx*vaMtrmtawocfaoatweuetdttbrbv^aeeeMtg,vi^mnaTdcnnnlruin^tdsTmgei^idrorPofuasb%stett(hbo.feieytf 12.4 gf(cUoteoaslnoetkeiwkccliieeatnHttderidaabsfroiMySrdaMihruxuf*afeloaenlnascattcreelifyidiDnetscdvisaarbnvtliyebbtaerUoat&fhtfioaoeaarnajrJtr(snenptgeaatratwneeadsddUaswUruJdtauni*cbtiuinmdia.naeijrpedngSacpyteilin.ofsrensIwnewindteiitaetiagtveaaheatdrirmnyacgpna5ioesl*e1fsc.0peuecers.xvaitekmrasmcptveledirt Patctsr* jPage 20 o f Exygen Research Page 61 of 110 000084 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Bxygcn Protocol Number. P0000760 brfaaftJMirck MHk4MMWrV009l7| CMetedofAnefynafirIbeDeAteWrmAtaLtLWtaCiI/oCMfFASei/MIrIf5ioEoTrHMOcDttawAdd(FFOA)isWtterby vsoeftrwtuareealyUabfncdae.atfoidvdeosemnoausingMuiLym33<orequivalent) )2-5 tBwenefdltsTtaOnpdcaarde#raepboooridietnsohtoeuxcldee!*dfaarUthmeUrdsdilurtee*dOenMdre,anAanlyyzeudm. ple*chat 13.0 AecaptanctCriteria 13.1 1U 13.3 wodMbafeIO1ftaeof3UrMmteeatrt04eiotfMnehnrH1#tuoikbko3nKaMdnocMseMroMh(ribtlrhwokoMeIoMtpLunnibolfadwo(unufaMHxobhPdMooeo9imHpeiweumelh4naM.v1otdteu3tltnIdkaeytf)oaeiuetn>wtkkfueepncocndlncrafdeatntpmiblrprtmaAheamyaeeeffourreililtune*hrhiPwwttcfeeemceFpeeuew&iOmeuterraIkAitmdexdhtiCehJhnrtahicyaewalbpevomipftdSueltfoOm.ctelevocuaaordtArcimtdbncgtiinadocoopiyeOfnne,taaidomtie(nhhbtdimonePea3e.xiF6tnuirbeidi0OeiexehecetiAnmwe.tfmpopemrwt\ueeatkhnM>if*Meeae7nonxLa0nItOlrmwr.ieuIt3eQcilhdtB0epti.iolomeSerii7itptthfhon0ilsae*e-if 13.4 AylibratiooatududtendtobeaitotiitieslouterbywingtheHuge eHExwrcrolwurdeTerader,amt.hemeeattoyntoabtplenexuxcmcelebudedre2do03fiw4exontrfat(chbteeedtcoectafeeilubnltaoulftmeicnbsteoritfioaibfdueixrctdraailcibttehradattioscntaonucdaldarvrbdee*. 13.3 13.6 2eRTtph4oepen9tmrd*eem2pioodri(meenRtoo*lteirmthtihMeGeeapiPreabcfitSoaonet)vww.fmfeiaceiItioefabndecetatoaMlGnfibjidemOraaasngtdIionrsitnaoraaernaeeadtbs&utbrMaalrta*lmdbfiphcublaeeeU#trmeeOmumnruveutetr&ayidetzentgodowdpt.emidmranrtitfioatbndmm,moiaturneu,dsttthhtahabnene 1m4aSnawlyittihcainlnuonthnelnytthieeislentmm.oIaftbrsetonsfiaiansaltyimueed.driftexceedsQtitlimitwithin 14.0 Calculation 14.1 gUbaetonindtbit4oanAJbplloeyewthkienagrMeeaa)gmuweLitnyiongnxttboeAewaeiteaontraedleeptrodrothc-aeurmvme:w(ulinmeaorfrPeFpOeAateffdraipuaMraminent^eLrs.) PFOAfoundML) slope xPF DFfbeterbywbUbdwfinalvolumeweedihitod,IfiMOHMiy. rut4on P a ($ 2 i o f 63 Exygen Research Page 62 of 110 O O O 08S Interim Report #23 - Analysis of Sediment Samples p. 63 Exygen Study No.: P0000760 Exygen Protocol Number P0000760 - loiiihttittfc IfilW itVl-- v- VGD017I adofAwfoiufcftoPifcAwNpAinLoHYwTIiCaAfPLorMflu1cTroHoOctOaaclcAdd(?P0A)inWwrby LOMS/MS 1 4 2 f t r i w f l i t ftrtU ad with known imounta o f PFOA prior to unction. vie tb* feOowlnj oy wrioo lo c a la ik*c tb* percent rc c o v a y . fumlMilyOfcjafldEmil)*mlytofaintmcongoj{pg/L)|^^ ui))tciddod(iiL) r* Exygen Research HpToP Page 22 c/65 Page 63 of 110 000086 Interim Report #23 - Analysis o f Sediment Samples Exygen Study No.: P0000760 Exygcp Protocol N um ber P0000760 ANALYTICAL METHOD Uctiud Number V00017II M ette* e fA mbriblbr tbe Detonalittloa of Ferfiupfoectuctc Acid (PFOA) m sou by LCM3/M AnalyticalTaatinfFacility: B3S0tce5lye6fRCoedAelie--coaaerea,*PrcDAhriv1e6101 ApprovedBy PTaaeuThlCnkoUealuLwel.yvider,LCC-ldSJ,tBLxy*imRteeerch . Dale / JV6ihcuePFliaehaeurltaytt,QpmtwfM,EayyaRe-- rob z s . Exygen Research Total?** 7 P a g * 23 o f 65 Page 64 of 110 000087 Interim Report #23 - Analysis o f Sediment Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 AiyiMkMMnfl MaftodNiHfcfV0QQI7I I analyticalmethod .. NedwdofAnatyaltfortoDManuaLrOiMWSfN/MfiSlumooctaitofeAaid(PPOA)inSoilby Ilia maJfcod a to to p toyad v tto iaalitka awl quantitation of perfloorooetMoic acid by Higb NifouuiuM Liquid Onpgiuofnpby coupto} to undos M ut Spotto metric Dilirtcr (LC/M&WS) is ta il 2.0 totay 22..12 ACproetonccaiyuuiittaiottono*earpvptraofpiriiaiitetoMraStIo33rybpenfoeretohtat.iidtinfanycbermralforpropersafay 1 0 SwvloJIaqmvaawnt . 333..J12 33..54 NSI8AArUonemtlMeymyiplurnmleiatpptcIMfiSaeoaUaiptl|.omcrreeefteedwMwtonttoipniMturhgoopfiatlrndongtuefomegtdrdhiedlydwtowoIrittlailxkttaooddonaibltaope.mafocrapieflloleobdw*e.iienndgtNotocnpaaiemqdupifUlbiabtgreepxitiretcofiooetno.iohmo pwf 4.0 KmgftiUtodtontatts 4444.3341 MAPWmgafiltmtatoro-onHiodcuodPmc-HLtuACoPocLfnfoCdtAiftrocai-ddA-.CS4tg.aRuo-Al<AoicihtGrt* 10 Indnumst and Equipment 3.1 A Ugh jwfonnaoce liquid bmoatognipb capable o f jHuuping up to 2 orfveBti equipped with * variable VoUmt injector ci^tble ot injecting 5000 pL onwiertod to a tooduaM* Spcc>octtr (LOM&MS). 5.2 A devito tooDeoi raw dtta t o peak integration and quantitation 5.3 Analytical *'***capable ofreading to 0.00001 g. M 50 mL dilpmd>La polypropylene mWfuge lube* 5.3 15 b L dUpotablt polyrepylm eetttrifoga tube* 3.6 Ditpoaabk micropipets (MOOvL, 100200uL). 5.7 llS'tnL LDPS a*m>w-muthtofUre. 5.8 2 mL daw HPIC vial tai $.9 DitpNtbkpipMH. 535...111120 SWAPuatCotmvpatSpae^nt)moPaam(HkaKVnHiafoGcldd0.0op6L(]agn>d)ClflrtSlO?OBpcLa)rt.rwidgiteh*t.opcaabk tipi. 5.13 Ultrasonic tu b . P*r2Pf> Page 24 o f 63 Exygen Research Page 65 of 110 0000S8 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 BayyMKaMKfc womi f ANALYTICALMITHOP ~ MflhodofAm'lydltoritoflmnniriniileoBmesf/tmefshurtoctinoicAcid(PFOA) in Soilby 5.W Wria-ictitinkir. S.15 CtblrUUieafwbWofipuunc50roLpolypropylenetuba 5000rpm, 6.0 araMftfnpblctyfMm 61 .AwdytiwIOoImbmkFtoophtHR>flCeyrtca$oiitito)2-1roiax3Qmm.? 1116,225 MMTO(BobenNbmttMil:leepEUoFSmntOtiwPT-:t(atA3imI0)i*J:CQ)iMweMtnAaroi ttOwinAnhtinWmr fin2201t121i.44.0v05nl 26622t5555A H 55 55775555 nfnotwLtoRwulel O000...33J5 W 6 4 tyeftttV o lm K L3 jiL (eie be taremed to as and) u 50 y U 6-7 Quaadliti: FeikAm-cMcnidtodaricdihntioacrr>. 6.1 RudTime -1 3 mfmtti, Uteatev* ecmtftiMi tr* intmded et * g&ida cod may Wclun^ad in order to optimise (heKFLCtyvfeoi. 7.0 MS/MSSyitam 7.1 ModrBlieboepeayNegativeidBMmode,monilotin|413-t369 nVz for ?FOA. Tto aboveecodmoee bo ktended u i fuide *od may be chuged in order to optimiM1bMSMS system 10 Fn^rtiefSliM 3.1 M oKl'tt* 1-1.1 2unsmaMesuiBnnmKaBoeimuiaomaceItDatOe CinmwLaotefrwI*p.epired by adding 0.15* | of Ataoute wlaroee m*rbeptepued. P|<2of? Page 25 o f 65 Exygen Research Page 66 o f l l O 000089 Interim Report #23 - Analysis of Sediment Samples p. 67 Exygen Study No.: P0000760 ExygenProtocolNumber: P0000760 Nwtorvoeoi 1 ANALYTICALMETHOD MfdwdofAnalyiiiIbrtbDetanninLtGliiMo3f/hMeSfleoroomeekAcid(?P0A)inSoilby 9.0 SU BtO fdfnp 9.1 fmndmdftek/FactlfleatcBSolution 9.1.1 VKpnftodeMinteof-100pffmLofPFOAbywegkitg10mg u<4ifttaannnlplKinila1a2n3d'attLriLfoDomMc!tbeodtftia*rpurty)andtatolo100mLviih 9 .2 amAlL&tltotMarottifLnL1L0Dfb0PiitgiEfirctnbtiLfoctailooirhiiittfio>nM<jfMfinKti)vAolfuempecoipfiflOdObwyitbbrnif*tgdaifttolfll A1.0iiftal. fortifintiooaniutfonofPPOAli preparaibyhrfl|in| IO nLof0 1 0 jigtaLMbiliMiofintivolumeof100 withmrinnelin 9AA 9.1.5 AloAIOU1LO0S2aXo5ailHLfLm|fio|jLfLieoftdDfLaiLLFDfOoLf&eofpBrfni(ctfbiokoffktoiltiLfqitiCjcatiaig*eo.k.thtekitLniiaollntiooiiltuhootitiaitno(aaobefurloffiVFvotFAluOlfmiipAmenjioiMvpfo1iNcl0up0mdtrwbeediytbotbefmymiaDbdOrinaiemewfinii1ihgn0 9.1. mTheetetemclklaaid1f2c5rtmiftcLeLtDioPcanbtauitotoaerotobertortd!rtrefcgeretoiu SppurncxtittBdMao*fro4f"pCreupdarat0i,tubkfi1 maadawmjmMof6 rotai 9.2 ftudeniCeJilsitkenSoluti 9J.I LeGaliWbitSiteMDStUcandlibmralsionprdtcaarndiatsiatirparetfpbartaitiIneoHtoPteLtCion*apireocr.edTuTreie. 9X1 TIdheeaftbimBio1f0tinvagmptli.lypicalxmnpie;additimi coocentntioasmtybe pipantitneeded. _TeXSC_foLCFetenm*t"uOac1iiiono11IrrM0S5aa00fr*nu2etMr_fenaaudbrtet_aroaettaotiitiddeaacPeatetVlriabmtff424e2w1riuot1a0000r0tu1ti000tt0wtinioeanteie.eKteoaFonfoSctdirVrmeataifornidenclun4444444.dAami0000000*Clfiemboerrflaft.t)rimttil itaCSanbntdCisDdewaadrFilrid2rrbiil5tartni50iimdmir00dI0aotiuttniiioaroinqnlreioa*trfiirtauXXXXXXXSC*CCQCCtdCfrteanmanmmuiHuonuilmBiam&bdutaniirdardidoddondnardddddddldtfiaiyt)yyotwni7dEya^y)n(**D-OSfO^44ltPEL tra*'* Pag* 26 a fte *>*"* v+! Exygen Research Page 67 of 110 000090 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 AKALYnCALMETHOD MethodofA-ailpsfelheDflunma*LipCo/MafPSMflc3fiiefMQtaaolcAddfPFOA)inSoilby ] 92.1 AMt--a/staMndairldr)ilxofkrMtSosDd.fretaswbbafl amhe preparedwithwh WJ U AgiSthdto2elw*unCLw1to1te<av*obChid.ipveadtoemtawiltoeaawtnioscanekaast.rtaadnodnasrdcsfiae1n3d-medL*pm<Jyypbraeppyriepiwaretudbes lOdk BitchS*Up ]0.1 tto pocRiEnfoaweentceoqfhtreeewodqMelruanrnaaebflowtmlietrycateeiOfoiiifeioaMtvodmaruerSmyplllt0teieodMekrrntpohfotla6deWwndblnaafbtebtlrccaodhotnrh.aDki(tsotouyrpaspyetitnidnctygwdaeodl$clynitcmae3cL0o(lsoabrarnIsdcamosl)bnptrirataoiodlnjstwpiniiidklclelbut)wedioesepaevicaleiefirae^ilsdyit HjO Sapbtoriin 1 1 ,1 J11l..23 TnAYenddedadg5eabdfdt,^rlnLtph|otoiorleuafberstelhaisdmaleaapnlouedlutomlattendixsi9wbo9eUunl)ctL.obopaoithtwyapnirdnopaaeycnlstiioMcaniescebfoniirao-1riff5loifsrm-m1tob5ueteaasm-(feitfreityi as 1(4 Bornintgrifiihinetvuobleu.me19 la40mLwithwaisrm the50eil polypropylene l)i CwxlriAifflEor^lminute*st-3000rpsxi. 11.5 CfooloawdJettdaabyti5tmCtl*omfHPLCrtwfiditwf(-(12gd.ro5jrntel*c)).bDyopcnsoatiAlegtIoDJmmtmLrmunedthryinol U? U.I 11,9 ELA1o5hlnuaumafdoibyLws(dpeeoistclhayamp-m5rop)pkm(sybaleLesns1lfeti0mgc0ecpHlcoleetmifce0oeg0^&opdmftaiueoybtLce.CosnC{/MfdinoiStaliioc^lnMwS$l)u.CrmonLa-oS5PfmolucUaa.lretriidngtoe.graDdiuscaaterdd 1 2 .1 InfectthsaaccauuztouKorcacbstudied,strap!andfortifiedsampl*lowift uLCdyMae8/dMsSaispyrltiaim. Aeabbcilknstandstri must praeedeandfolowal 123 SiMtaflnthdtiB*ioilisPdPaOdAiaouoktfeinaipiyetoiuitalgi*tot athastfiveormorsconoerruadonlevel! 123 Ablaetnegnohpuowttrutetdibt1ae0dCWoaftMothaoeenveaefnordyd3o-e1f0a*tfatmlammlpeplamt*sstAaenit.duardEdMxtrmancuiatseejvdebs,etiaaInndscaalurbddr*eedismeaitaottfthbe* ftp*') Page 27 o f5 Exygen Research Page 68 o f 110 000091 Interim Report #23 - Analysis o f Sediment Samples Exygen Study No.: P0000760 ExygmProtooolNumberP0000760 DntiiNwcb- MethodNinberUMWSi A N A LY TICA L M E T H O D MetedcfAnafymfartbDetecmmeutJoznhoiftPmNflimtwGtaooieAcidfPFOA^inSoilby eJ(lboxDtraaeccwctaeoddunbctyfatertxfttioraafceateciodncdtaotnotfdbtarodaeftkaamttrSaytcatnebddoasridnta*fneincdttaeerdrdtps)e.tnIdeoedeUbvemegtriynna5ain-e1g.0ooxftwnic^Ukwdst 12.4 Uh Kbmt(antedtwvflt ft quantitation. UnearMatedcurvesik rstomonftuwraatcroedaslytbsetnettmeia.nntityetnetebdyeImenocawnKnagtieFaVuiradnu|sMingseIfeeLwyntexf3b3rin(goroefqpueivaatlaerneta| )ZS te w p ti t e -- d m ld m l exceed e n te il o f . A ny sm ptes th * t u t e ittafcrdT C ipeaec* should b e fio te rd ih itc d an d reanalyzed. 13j0 AcccpUix*Criteria 13.1 13.2 mbodCMalfutieuna4stthr1ktfaobStDadeeianoobwntbnleautiapn.imikn(ia3HseiPuddmiFNPauOo4iistw1daAt3nhtuueoem)ttwI^sccuuvouaawopttpeeaismerieeaoamkPttotmcFeofoOlu*aOtrsAtedhtHsbaaapeuntoognl3nehfh0dcvteuae*nrurltgboiso^oigcU(nnhmtodoteditoPee3.xPnridOtaheaAan.mnemwuthofberoolaLm,nOwkaQhspailemtrIstpfhaleei ' 133 RacewnteoreoiteleptkMandmatrixspikesmustbebetween70-130%of taenatreberoewtonfvitaoluyeles.sIsfhaocuoldnbtreolrqo-ekx*tnftclttetdo.utAridneydmeaterioxcatppitkeeblaoulimtsiudae.7th0e w13a0rr%antsehdo.uldbeeviJuajsdbytheanalyst todotermineif re-otbaotianis 13.4 AnycalibrationltandodfemdtobeastalLitlCftieulBerbyuctngdeHuge * EHorrwoervTeor*dmetyotbaelneuxmclubdeerdofAoctnraothtaedceaafckubUraitMiouoitfuddeucntiKtbhmatiocnoucladrvbee Ienxjcelcutdeed.dtaut bmexceed20%of(hefoal rusherofextnewdstandards 133 Theoooela&ucoefficient R)fttr taUbrttkmcurvesfederatedmuit be 3KfpXi.oWy2ak(Rte2 2na&p9eK5n).unJfbecatJajtkeetaniatuoraedsjuulst*t finosiltruomutesnidteo(pheoreastiolnim, iaun,dthtehne ettndaidtorIheidBvmtsetofsimptoiteutdbeleenilyaed. IZ6 iR4e%tenwtionftttiiimtaentabneatwlyeticeanlreann.tIefrdvtsanntdtoadmmptcderimfturoondostfdtUrifttemfotrswitthhainn nSMlytUeJrunthenheictmustherasrulyaed TffttoT? Paft 26of65 Exygen Research Page 69 of 110 000092 interim Report #23 - Analysis o f Sediment Samples Exygen Study No.: PQ000760 ExygenProtocolNumber: P0000760 IinnltMuiEb StatedMinionVOOOI711 I ANALYTICAL M17BOP ..... MethodefAoibwfi*IbePfatweLriOnMoSf/PMe9rilucioocttnoicAltd(PFOA)inSoilby (4.6 Cikuladooa 14.1 bfUtaomtead6lMmfepbBeyeotkbwmelnlAfo)uqunLafyilnknfot(uftaoStCmwTnkinduvbptdomApwcvnfoc:a(cKunnelwofrFeFjrDs*A*kfwinepdtm(uinnenlatrfL), PFOAfound(naff.)' xOF DPbcMrbywhichIbefinelvolumewe(filmed,Lfneotttiiy. 14.2 feFeorfomlonwpinligtAiqrnteiJifIntdnwtoiefeekbnuoltwfentoim*poeiurcneinotrfePcoFvOeAry.priortocxtncion,um lmvoy(K)a [jotilwiliyfefamadtna/L)- mtyttfbuadipcontrol(ayT*)] ioafyteadded(ptfL) 144 thelbfalowingoquniontoconvert\benoun*afPFOAfoundinng/Lic P^iiPPbX FFOAfoundfppb>*IPPQAfoundfart,)g ntradediQ-MU) aojp!wtoghi(5f) W.4 bUamteddOienffeeBytVwfjtnaggfaelquationwcilotlMdieameustofPFOAfoundinppb PFOAfound<ppi)dbyweight-PPOAfound(ppb)x[160%l wildeofideftl] foe*7r? Pag* 2? o f 65 Exygen Research Page 70 of 110 000095 Interim Report #23 - Analysis o f Sediment Samples Exygen Study No.: P0000760 ExygenProtocolNumber P000076Q AINMAhiLxYlHTuIBCbAL;MVOEOTMHTOOD MHImlif rtI)itilii i>i tMiiitiaiBMfctbyilLmO..M....S...M............. m in i 1'1lil (T r m ) ln AaslytjtaJ TWingFacility. B3gt(^UgtCwP4iRwl>wwgr>b,PdDAiriv1e6901' Apffovgd By. C--iL1_____ PTttucltmCoc&iloioMydcrtLC'MBiBxyjai ofaeewtb a /m / b l / f/VhfkeDe-PFrluihid"detym;Optntkm,fttyg*Jtieeereh _Dm*titiuJaL D*t* TcftlPite* t Exygen Research Page 71 of 110 000094 Interim Report #23 - Analysis o f Sediment Samples Exygen Study No.: PQ000760 Exygen Protocol Number: P0000760 MatedNietervrooiHi | WALVTtCAUIBMQP ] MethodofAwlyjiifeethepf minmignofPoflwinwwtin ioAoid(PFQA)inSedimentby LUM3!M3 1.0 Stop* aSIVdpdreibmteyoMuHtirgichitDMPettiberibectneoamre(LpnlCocye/MedLSiCqMburSitdV)emClahroorodfcmttaioaetBoangLrdapqhyuincotituspbleodaotofpaertloaoncdoeoineuMnoaic 2 .0 Safely . 122.1 CAikhwnUytohbewipnFreCepamfetlibMorSeDtoSytaptraarcaticbeu*r.odbieanychemdcitbrproperlefely 3j0 Simple Xequnaueut 333.11} 33..54 NAMAdpSoirwUmiowleerjaieeppaemctmliceKnteppiJuctleorOetesplek.ojremeeiefoidtiutoeiRlnwMrbatpaMeroiiigchtpeofnkldrertufaeeerveddrserodNwithibryaioirlmleucbeliidiMeadiBbubpeee.ataocAtiedfileebodrweaimenpdiegdie.iteoasinaepmqleupdilliinbfograreplevlatnrluoftoiMrro.tobmi* 4.D ReaganrodSroodMda 444...123 AA AAWMmceeitmthteiac-anAHroclP-idHLA-P*C'dRLgdeCraiagtgtereroa-tdAge.rCa.dge>ReagMtfOnde 4.5 RorftuwooetiatrfcAdd*Sigrae-Akirich iJO lotrwronwdquqHiMm 5.1 AhighperforarneHqddetamategrapbeapeblaof pumpingupto2 rpoLtocrcenneeaqcudipptoed>tawnidthemvMaraiaebBlepvobhewaueieti^ie(dLCo/rMcaSp/MabSle).ofinjecting5*200 5921 S5AJ 5S.A7 AA50admeevlyilcddectpiolVOceMoliblaefocdepnotteplywpiNdroaepdoyiflfreoenreepdeeiaurokgtiratioftoe0gg,0rlie0a0tit6fa1*r.go.d quantitalioil 1Dtd1iam5p-emLetLefi^bLolDeamPitlEeoefloaocppripoocwlye-pemCrooJnOurt-htiQrotaOeticaLr,oi.i0nCft-i2ieOt0uubLc)e.. 9.1 2mLcbeKPLCvialkiL 55..010 DAinpettpefeobelettwjip(1d0c0tm*1.000gLid 10*100ptXwithdhpoeebfodpi. 55..1112 W$?atenre*uSuipmNnkroViebcU6.ec(lfl)lClt$P8 cartridge*. fee*}n P a g * J o f 65 Exygen Research Page 72 of 110 000095 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: POOOO760 Exygen Protocol Number' P0000760 i ANALYTICALMETBOB ~ J MethodofAwlyihibiAtDtfcmiMfaLiOoMfMSb/MtoSrooctmicAdd(PPOA)irtSedifrwnt>y 555..J11J34 WVCwrwirtiHmiriwfbr.yMtoibpotohrl-tofipuuof50Lpolyptspylgtubwi300Qepa. W Q w k y p h i c Syrtam 6.1 AniljliciJColumn:Fluopbu*RP(XayitoeeScienpfcl 2.1 nun1 50mm,Su 64LU..234 t<TMMhMaoofboMcbilpe2ninlFi5t1tfaMuthntmet4*f8<9i1Aw*)3>C:;[:Q3MmrthMmAomlmoniumAeetiUiowee FlowRete 0Ij.0D 22902Jt.O50 26U5 63 22555 333773L5$555B rinlupini 00000.....33333 44W..7$ QbRwyuwmnTiMmiiiueV:ito.-luyPnmeook:aAuliilimLe*tu-.aexbteermnalcitraMadevddlPc*aXflrm*tkuucAh*ir*v3Q>nL). Thethtveeooditkiittiebundedletguideandnwybechangedidonfcvto iTilibiiTe Itii ITTI T ijiiiim 7,0 M&MS3ytfca 7.1 FMFoOdAe.SltttM^yNifriveMJOdmade,noeiftoritig413-t 369m/zfer loiwptaibtoow<meoM3dMhkSelyieertecmia.toidedti sguideandmaybtdwgedlnder 1 0 fnpmttooofSeltiHaiii 9.1 Mobilita . 6.U w2wmMMaiuemaaloeaettaaqtttoKe1uQaC0eminiweiiiwiraJteirp.rependbyadding0.154gof Exygen Research Page 32 o f65 Page 73 of 110 000096 Interim Report #23 --Analysis of Sediment Samples Exygen Study No.: P0000760 Exygen Protocol Number; POOQ07SO 1 . - ' ANALYTICALMXTHOD tfttbodofAnlyciibrOrwuroinctlimLOorMP9dMAim9oocuioicAcid(WOA)inMimratOy 1-3 Bund!Mde U>1 l1OHOcDmeniLIbelafwidrt(wA.nuticps^mdbyedding10 ml.oiaceticKidm Aleresvotemaybaprepand. 9.1 SaadariSlBckffatSiMfSdulioc 9.1.1 9.1.2 9.13 mAmPAtftMa]fgrla0Oouaa>1rbfa*i5oihj*faIlm^ntcaifnaaLe1LoiL0arfdt0Ditu1Mtkia(P2ciHftNBhLnannllbiinCLLoslwtf*nLtMk4hW.oonadfaw-mtuIOiftbtcthiiOaidttltotlottAalp*aie.PoihfpfinPPnuaFOLrlitoOyvAo)fAPltuainfldQpiapnArotpHpbfvAuy1*r*0dtwdt0obibwgyHyhbithBhtrrmiiomnimtginlIOi,Qrituwgfai)nmIh1De> mal2L9omfthLtLiODPpBtfmboLtt)cfo.totimtoaAnalvolumtofICCwithmeth*miin 9.1.4 amA]02L,51onfntXii^.gU1L>.0f?cyBrgtbiffaoitaitLt]iaiao.oohattwoototokaiitfciaf&PifvOoluAmiaeimofy1a0i0fwlVityhtranwtthinutgol1in0 9.1.3 9.1.5 fTA1ab0MeMefkbloLalcnikiooftwfian1aLb4)ot2afe05t.vmi1ftieULpiLklifoarOtatitfLaBiobsiLootalifatlUitokia.io*ti*voteofFtoaFbEeAsMnislovproarelulipnmaaerere)obfrfyigIbWerxinwgwirnitihgt aItpwpowtabU*imdaistotyo4fp*CnpumdtaaonaattbkBbrimaiawnperiodgf0 month 92 StredariCalibrationgototient . 9.7.1 LCM&i'McalflKtiUitiededae-ptaparelinmethanolviadilution 922 HefwttwfoOloJwingit typicaliwplaiadditionaloowtotniioiumyb* WoQfFnuoiMroiAtlUcnitiuiWka 1wto0oo0 510 2 Vhum (15 . 2 10 10 _J2__ DO(LtttICo*ooOt-OoooiO>dO __ wo CofnawFnaitnoOssa!Lltlow so10.0 2 .0 031.0 0 .2 Page 33 o f 65 Exygen Research Page 74 of 110 000097 Interim Report #23 - Analysis o f Sediment Samples Exygen Study No.: P0000760 ExygcnProtocolNumber.P0000760 MvlMiNu^w VOMIm ANAUrnCAlMETHOD MethodofAmdytfitytb*PWnnlnsrimofPoffluoroocwtoicAdd(FFOA)inSedimentby IMS/M8 .U..J SnlorrcanlUSMTlif,enuhpotBedMioimolmadh# iin IZStbL LJ3PE nanvw^oeuth boniet U i AiwnMvehinufndeoAOhadeM oflinM amftybeprepedu ]0.O 10.] wEtchMtaktwd)fcodfefeasnaphfeeeletrnedctetadp(tyupnictfaMlltyed30oooroJbimol)imfoonilifliendcludaet k1nbowunt tO.2 RibniebthfmBeOwquWmebBIitmWymttQftuiernbficeoolcdn*tproofcdluiUpUfboaoreOn)tMdeitvpygriloutjfpeyhcptcrMocHedaunradlrocoverwyfiollrWtfnspheactihre.ri 11.0 IL1 11.3 AfWmtrodad-i4ifMdb0.5mapJiatieuoOclfef*rIo.UtmdtpMsloedtmlmelowba$wd0e,mcl)^.L,pvoolyipterorpynldenIbetkceartoriFnuegwertiu*bsaeti(ofonrdtirfnykeur y-"v 1111A.3 11.5 cCCJhbbruooUtdbeDHi.fiiwOpna1ddfbiceeyC3lB0bmr$oLJKoftfKt3an0rCQtriwd0rfWpwmr((d1"'2tf-gd9.rQ96psmiubmeL)eitw)b.y.DpouusointlgetoJOoLlmueanLdemr(uiteinfdtdnuruQyulfei sunglsonbeconditionedC-.i S?Bcmridge, Discard IM. AMdOdtr2iftdigemtLaob*fmCeadp,mvealrielodtteaddsbimekeeso!nlefet iwsrtishteabcotitotonmshoikfathrefe5e0-m30L ' . 1H1..7l PCcsciatBruftahfe>mlhuehteebiomloitn-3to0000iwpnnicfeETP-3E0ciaarthridMget*.*.Colectft*ehuta. It.? WfaauibtoteheolcMotbardaw*ewteithp4I].mL, ofmeduned. Colectdieelma*rod*ddinothe 11.10 11.11 fQSAodftotBdoocwd0wi>etb*dtrambideyafeA2ro.0eronmoeLldtooCf1-H21S0PP0L5CmMwLsrtetreidfrw|(B~a2(tetdrraonp6d/soekieinl))d.bDoynopntnhroeotusleiegtcco1o0nldummcLonnmrdweittihodenrneyodl 11.13 Bhdftwife-4mLIDQHmethanol. Colect5mLofehtaiintopsduied 11.13 1A5aiQaLlyfMGsiyapmiqppley*ttBirionsgmirleilcifiefoBetpiirbaaycL(Cfi/nMalSv/MolSum. e Sml). r**;on P e g* 34 o /6 5 Exygen Research J k. r\.$i * Page 75 of 110 000098 Interim Report #23 - Analysis o f Sediment Samples Exygen Study No.: F0000760 fxygenProtocolNumber: P0MO7SO tl--irb IllOl)rmtarV(KC1712 I ~ . " MtefcynCALMITHOP ' | MWbaJcfAn]ylftrtbPwlmnbtOoMiPSnlMAwBooniaoicAcid(PPQA>ISodirocniby )2j0 Chaenutaptyliy 12.1 LaIqnOaMMlyzSefeAde4Ht3omnpaelyeiaetum,too.unAtocfaXcfberafttotocruin.daarmdpmleuasntdpfbranctufidaadaanmdp)folionwtotheIt 12i aSutinotdboeimliodtfaPdaFdOAueooDwaybedaudailna|ilto. tiIea<liveormoreeeoeeoiritioi)level* 12.3 biMAklereanntwgddetanaMenrdnttaioriomfnawoaonyadytnboaMpotfmOOinaojewdmcenaetnepdtdneddoayaftc.5iQa-Alt1iaob0mtrbaaoMptnijoulenamplmnilenncmtag.nM(odttoeafivrdadai.aacwicanotdumfheonmltomtifjwbo*ouremtdainbcobmoluycBfdmccCeamaCdKl!ifbpbamrpraatatiinitoohconnedf amodeattrodii)d.<IanmpAlteearccts* eaJibmtosmdanltnunboQilint ud tmt *' 12.4 125. SvftUeeamopnfmtoMaimapnldiiltinettMnetetnataafaluibyrbpdanotraataod6naanolaotltedtaiaahimnptaolalayuoudtledweaabnvnlayooCbt1t1oMmfeaexCldrcaebqenomdtrpfttinairoorifatittcntauttrorioiidndniMte|rloMionpldiatnemalnefnarLadeywrranw.oxitnag3aAnM.l3dynaeiyn(reodfsdrom.enfpqpteurJvaivteakatlaetbranerntae) 13.0 AcceptanceOfteri 13.1 doCa/bu4gM1h3taemtrtt0uo9.F(37m41)9tua4um1n3od)aaiqmopurwepeaaeptnetanaattbckoaonlfeeaaup*aoooiffdbCeiatroibMo(nihaodt3iPn6tFld9OeaA.inauniOontx,mwahpileiyeitni 132 bMlaenthkodOMbtOekkiaaPMFlAMaltleMwdrienpPePeOuArKtbaleove0l.2pneprntaoLr,dtmhenthaeLnOawO-blaifnka 13.3 MdJnButotDirop*invlessoaeietwwosntfovbaaflcuooebnatfa.toinLIiehfdaoUauonnloddomtbhaaeenoideraonmpHtiikSraeaurtmrxMetciiiacmUdo.teautAnbmdeneyumnndeiubexentnaibcxeecteetipwpdietk.aeebnleo7ol0im-t1aii>td0t,%7ih0oc.f w13u0%iflOadbo.utabecntoModbytheanalyaltodetonateif ro*curcicnit U.l Anyoatnnattoatnxtartfan) tobaaatatieticeJMillbymlnftheHup JEtorrwoarvTatro.,tbm*aMyabei nounmckbuMoffaonnribtaadocaatoUulbartkemttaoitafndardUaRtKhaSttioonovciudrvbee. UKgaelatfttbdd. matnotturned20%ofthetotal numberofextractedtindifdi Hi TheQMialalioncoeffletat (E>for oaUbndoaenrvofanentedmut be i2tm0p.9r9ofa2prrdi(Rit3eSAXteeQprJseVtemheuuMetIfbeettiointfaakateaentpalba>eraihsdoujuultxaltdiMbnealtrreaamunUaalntydtzeeodtph.eesraetbUnm, aiun.dthdeien >Miof7 Page 35 of6$ Exygen Research Page 76 of 110 000039 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 I - aWALYIXCALMETHOD | ftMhmtofAmJytitfixUmDUmamMkLnOofTMWMflOyosooctnaicAnd(FFOA)inSedimcniby 13A R4ttmHwfloitibiftchan*xaiintaaffyMdcMalTKuaas.dffatteibieainlqdnudmmpeiMdricftmcateeneobtdAriifttbmririwiUhiuini msaailjffieil rm ftw fiM w t e uri bv nanlycd. Kl blUHamaiaatdhileBblbdpUbtprtkltwitMm#M)nwlm.)ioiX|iioAttfitJWitpadaNdt*uopdftttcpu*arvm#:o(u)nkt*p*fPrFleOgArfou*n*dp(aiorfnttgitt/emnL) ypnAfatwrffnyAwn.ffMbarM.lfilTcqpf) pp BF-fixtorbycflta]volumewutiLutad,ifucMury. 142 OFoarfr&oangfttxqtnlfeisltdanwleboblenttwlMafinopimerel*ftottfroPeFoOvwAyp.rWtocatrttfioo.me Ronwy(K)- Iw l-- dytofandfrylnL)- BttlytxfoundincoatreltBgfaiL)] xuJyteiddfld(agiL) 100 UJ BU*tfnet(Uffbfo).UowinfquetkiioeocvtrtftamountofPFOAfoundinnpfaLto P F O A * fPPQAJhimJ faafeaLI final volume ml , wruptewdjbt(51) 1A4 Ufamnd&in*ppbUdadqnufatiyticws(iif|bnt.ooatiiry)tocafoulaMthaamooRtofPFOA PFOAfound(ppb)drywwgb- PFOAfand(ppb)xILOOM/tot}bdif*.)] r^T*r7 Page 36 o f 63 Exygen Research page 77 of 110 OOOlOO Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Exygcn Protocol Number P000076 AMNtAtiuLdYNTuICnbAtLr:MV0E0T0H!TOMD MUitOfAttlyitabrIMmluilODol'MHIiHracctaiwteAcM(FFOAIInFM M O C U nibyLC M M W AulrticalTm!Ptcilily: BOSitWtygCIleMotUolrnn5k,nPDhAiivKcK1 AfflWtOBy: - ^ J s . .CJ L____ Thll*lCk*oJioiMollfm , LC-MS.Sryim JUMw OltB /rVJVaUykhlb*bbiPHFmWm*i wld?rtfytit/,Q&pmtffa/oixygnjUmicb DJb>*4r Exygen Research TbUlhtM: S Page 37 a f65 Page 78 of 110 000101 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 ExygenProtocolNumberP0000760 AftAbYflCAtMTniOD MelhedofAaalyrieftcflI>t wicCitnaem6oeabqyLfPOtMofl&ooMraSoeuanicAcid(PFOA)tnFilfeand 1.D Scope UmmafbodlitoWiBifliijiiilbribeindiripnadquantitationofpaflsonecimic a3dpdwdbyBBHi|JjhDiFtrcteri(bLaCmWmi<Mliq>uCid)iCnbGraebmedtotfrleinpliiy. coupkdto tandemMeis 2.0 Safety 3.1 Ahutpaehenvarafclihwlorypractica. 3.3 CoonhlfaeappropriateM5DSbeferebladinganyChemicalforpropersafety 3.0 Sniy InH q u im iin i 33.31 SAuiLrafkoHitOadgdefbieeiiperencaebuleafdebreefxotrraecetixotnra.ction. Pin*lbfoiensamplein bawoidbdliIpnnurontktafoiwfaSioedablIwabnna)mcigpSinMetitccrtahwgeittaodmvryeplneteige.hinlPtltaoncueamtlhoewsstaofonrairpgleeceauribnnotcnilolndimtioaexinidoe*f 3.3 laenmalph**coMwtionprocedurewillbespecifiedmthesamplingpiar fortKia Wjwt. 4.9 IlMKDbndSundmb 4444.A..132 44..56 44.7t 44..910 444..M1U3 Watar-HFLCgrade ACacrebtoonn(itLri2le0--4H0PLmCegrirta)-deReagentgrade MSielicthaagnaoll-(9H0-P30L0Cmgroadbe)-Roagnatpede Roriail(60-100ncab)- Raagmtgrade LSt-~OuApeelcanonkcoaUlac-LaHCcPi-dNL-CRH2egar-aRgdceeangtgerpadigerade DToimbtcefo^-JRdkehagkeranetgilraaid*e- Reagentgrade AnuDOdumAcotst- MXB.ReagentOrade PflrfborooclinofciAoid-Sigma-Aldjich 5.0 InavMMitiDdRgviyeMot ' 51 55.32 AaAiAldldvhareiglnyhittat*cepb*qeteuotifepttetpmilneedie*wcrcae^iwfuelWiqau#viad*ftorfianrcbrphaler^odmviinoa|ltutteoomgjrDrera.DiphiOahdOAeOccLataopgn*adcbqalepiaaonblfteitrpoiiuefmainp.jeincgtinugp5t-o20 2 0 PgtMo/65 Exygen Research Page 79 of 110 000102 Interim Report #23 - Analysis o f Sediment Samples Exygen Study No.: P0000760 ExygeaProtocolNumberP000076Q ibw > iMeiMiBbKVwgm] I ..................................... a jia l y r i o i , l a i n o p " | lbeflio4ofA--lyleimiwDtiWfliwluiai>ftic<lMoeqwnftfcAtwt<FPOA)iePithmd ' OaubyLCMS/MS SA JUfyiwyimr. . 9S55j,4S67 151T5205iwwmnLLmLddpiiiamyyuo.r--ihtballipppirolrlHiywy--ruoIrpyryyhlmmccwwnrirhifl|igiftuubbce*i., 54 55..1110 2Di2mSiiLy-*QncLliMrit-lDriHrFaLaCnrvoiwaJ-mt<iiS.o0-fl10b0uoLtt,iulO. O-2 0 thiL). 55.4125 AuMfipMM 10-190M.Xwithdapotabktip*. 55..1154 wSPiBtiKdwttaa(2ih0ankLe)it.Uqwfca ml na 10057177). $.16 CitirifcioapiMfipralnc$0nLpOlytffopylnMbU20C0ipm. 60 CbvBMtofaAie S p tm . 6A <tntlytlfiirtitm Tm : Fluophsse HP (X iystM Scientific). M m n x So mm. Jji (P/N: I25GS*OS2130) 62 TeapeW W 39*C 6.5 HolePfaiM (A): 2 oM Ammomio Acattfe w Wler 64 MobiMPhua (B): Methanol 1.5 Qndfentfrofnm: Tim2216120..0f.09.5i0ria) 364$A 6255 25 6$ 337735$555 PfmloUw03mRiitrei 060a-.3s33 iS iMMhohVhhMMt l$^L(eabeiKiaHdt0i4akKhii$O^i}. 6.7 flwimiiifomr tnk An*- ttttmaJ atandMd catihradoe curv*. 6 1 R m H n r ~-Z3aunutaL TObjtimabizaavt*iacHePtiLtjeCot*yattiofiol.tesdodtsfcfuidetodnuyb*(tankedinordrto h|r}fH P a g ti9 o f6 i Exygen Research Page 80 of 110 000103 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Exygen Protocol K utober P0000760 BiyfmBywwfc Method)MrVwwirti t Mahdo(ABS*ysUftr4*PcA4WwiAeLii*rlnrMCAfJI^*fMliMITaMHHOaPiKicAdd(?FOA)toFih>nd3 ________________________ Cbm ibyLPM S/M S 70 MVMflflyitta 7.1 PMFoOdA*.&NMlEd*y}|*gtfV*MIUkltt0d,meBftaitu4L3-tJtmfcfor ' qTh>efaibmoi1^hceoldodSUMigS?*}rtiaiLhpdedAI1 guideladmiybechmydwouterto 1.0 rnoMdAOfMudoi t i MobilePhase 11.1 2exmaMeamhmmmftnihmwtmo1MMiomisLw*ftwaaiterp.reparedbyodding(U54gof Sr2 -StationSohttka 1.2..2. 7c0cfdiHdinDeotiOhMOydldrtyaicdtfdocielfMmUoOOfMMUlbMliMalie.toteiflntoiltuveonluemjeporeflpOarmedLbwyibthrinlogluinegn*7. ml Atomtevolume*vaybprepmd- 9.0 SiandvdPreyedm* 0,1 SuadirtStodcTcrtiftcaonSolution 9.1.1 PnpmstochsoJodnoof-100figrmLofFEOAbyveighing[0mg orwfM&rtetJdyaluia*1ls2t*oHodtLfdL(eo>nf>eIcbtlodttflbe.fpurity)Anddilusvso100mlwkh 9)2 A1.0pgrtuLfixttBcxdoDsohdieeafPFOAisprspuedbybringingI minL*o12f5wttLlOLOOPpEtb^eattoleJ.u^toAfliMlvotomooflOOwithttdhsnol *1.3 A0,tig/mLftriflcatiofiertitkooofPFOAi*pnpindbyblinking10 rbLofthe12)[igfatf.sotuUento*fiot)volumeof100rjtbwibmlin .0.1/4 sAlP021a1ljLiftBtamLfteAUtnte*idQ&aofcttkofPFOAipaperedbybringta* m10utbelsLnlplaMAbe1250.mJlp.LpDfaPLEboolludt*ie.nto* flail volumeof 100 with 9.1.5 TfrpotopmmmdimicekditoaCtoyoA4f*pfCllnifcpeiirdtebfmoncio.ttlaetbfoliiiIbarr*etuoxbiiBeuttennkdpJeroio*ditfoli|fvnmioonrthsi tw to tt Page 40 o f65 Exygen Research Page 81 of 110 000104 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Bxygca Protocol N um ber P0000760 MetoedtttttoVOQOiru I NAUTICALMKTMOO ~ ~ McfeedefAaa^fectiwDMcaiutfetofMlunoetamcArid(PFOA)inFtihwd ClunibyLGM&MS 9.2 StandudCilfenCnaSriatkee 9.U LC/M8/MS *UbratitttaiMdrdi k* piepend i meOMeioi via dilutkm fd c 1-9 pfftail. M fleidaQ olutxuk. 9-22 Hw futfariaf ie typte! extntplc: ddttuxud Mcentmien may bc pnpeed needed. S9tCofJPl.umJoti6No40m041n6114i.lS0..1.f2f9005atrtJf5oattovUewmLlnU)eeV&fborj211nn111.t.0.u000050&tmtBBehsdDini4lu111kt111mot000000e00oiU0000dst12S*mLCUO>lu9KO00OPf.s09MDtJl4OofB0.Me900S2Os205n255nfIr5l5LletllOrruAw-enouthbettle* 9X4 AtJ2W*CmtUoY*eClukmeepetldixoeeieottcQeist,rttkmecftfXJird*nUybepreparedu tao StockSetUp 10.1 ccB0otno0nhnIbiitduncrfathom<nslf(fcebtmocMpottooniltroalxnipedUctotwd)ot<otyywpirktrtf*trollypard2o0cederunrlaatruloreel)lomofevuretrilfyliaafodciruihdKebukuscoLhwu.nn 10J liUn^0uaqmulivtyittfeenrreSsefiiPdmptdaUfeferoIcfcfttoiivpyeqfjcepri.lleitiaandpftuwillbeipedfted 11.0 SimpleBmctioa ll.L Wtifh Sf effroneinopllato50mLpoljpicpyleo*contrifu|etutai {farfifyuMfed,npketfdindmixwell) IHU3 HAdiedv3O0ntoLfloATeaeeteotoeifrfeeBeteRdVvb'-lilktoeioira. ewrlnctimihiltcrfcr-13minum. 214 PiekRndeciMttiofltNSFBtobteeediOuDzaitopeer^hepednaefc. 11,5 Pi(kthe20itoLSPBhtbeeist4u.i. wdb2f f}oriselk7 $ itlicepK2g tht<oba2a0, stL4 e1 f9BLutCa-iuKtHh},. .CDonimditdionlUttw* funhitentr.imDowisfolvt2if0comwttnh*f emeeiutmhwnoul>. )14 S*jr.Bil Hw 125 mL peiMhyri Bukt by jfeeiag Willi the 30K - . rdfeiitmuhevtrtfnbtydyldebklyMlilnttuaocytsflheIlfmMertclw{iiohtthraedi#umctiamamee)o-ehiftDakntrttry-adfKeieintw.fleiethhetOeoemdtpJUtobelytobleemfeereenucees,, ' P a g * 4 ) o f 6S Exygen Research Page 82 of 110 O O O IO j Interim Report #23 - Analysis o f Sediment Samples Exygen Study No.: P0000760 Bxygen Protocol N um ber PDOO076O BupoJtm ta MMtedttUBfc*VWXn?J3 1 . ANALYTICAL METHOD | McdlPdofAittlyartfw-ibBDciCCfrahaiutbitayotOfPWttflSao/MtcSwtquukAcidfflOA^nFiihifld H.7 flfctriftjitee50mLyofypwpyloetutacocteiningampim2000/pra 1l-t Dhs*este&efutafl*wkf.ttoColteevtidtlhtlcneslduiStPBin<Doiteam|2n3fimtteLtidiniasmideoc(hiepmwoihuathpoef 11.9 HAACddikdtttkMit.a1m0jamitiaiLirthwfHfbtco~feQ1i0oambf>tLriplcotoflesweIewbtet>emuba4imleitpitnei*tnt(ohettubt#urbe5t0.m30isetccoauirditfuuijdfrtuinbsee. 11U11...M1120 1UJ DufnCSt>rleentrM-enem4lew0prififefmtIittfrfaMeftinta-sAuUhtbeatfu*pl5.ief0adiiem|oiBwaLaatfospebiroihJeryn-p]mdtOrohapcgNyou.tmctSrimebtPainEtueMcptaaawluw.eimtroiiott.tiahl'inaCcmt&is<jlt;enacrantmiiAapkfireeeena.luitetteh30ia0t0ionrdpmimaei ILIO Pdsa*tnsaa2tin0onmp.eLadrf^mpteMdrfilriiilk.Ortmjh6SPBoJum&sndcombinethelusiin 111111...111756ITAMetdueadnifpcoaeef3anaIsh-ci4-eamhdtauaopnreeplddtivafnioaettatraismd1mep-Ho,wFboimytLeuCaoidtluiv&liioionaagmgliA(2aeisrm/odintLtaxiietdrieyalifhoptvtv%i,aeIpntasaaf0nepKpontprpi(etmeatems.<c-s4idlu0pi"nCed)m, eDttisakmeinedo IL.It A adya -- piia|taoBO((peeyLQMS/MS. 13.0 Ouomatep^fay 12.1 13.2 12.3 ILaSmtWqyahnCtajeeeuanMtlseetytdfiennbtitadSadde0erdiMdeInMatSactrAtaalfruqPteideMtphPetfteeesOdeeia.DiAmnbttaifehfl.paisgenfedmApttnasi.naiutrndcafmiSavtaiestefyoelictb,ntneekbrddsnatfaeliiatttraau*taietmfetemstimteriiaediuilunte,isJtdssebibtaitstDraidnewifnpiivnftrmUeeecurdlnsuuloipadbrtebidenmrydpstfoeroitooredcrscntaietfsbiildeieebeedntrbetaien*iaetieminodp&dnsrptairwttAlieiiribhwilamnionjnywaahadn5yvad(-enbhalJfetleiiil uInnttearmsspteeartsn*e.d, cVaSteyta3ti-oOassatmenpdleasrd(mi muumsmi bmeffotraEsettcioannddnolatoufsintajencdtiaornd!}i.nina 13> Upvseoasfretsnwilaaaitarmceeasivybfasbtsretcdemint.ataantiaeelycdtuearrbvdyoeLeutcecMrernqrMeuptarieanaatilaoisn!tau.isMinLafina1efsuaLyrwmteiojg3d.h3wti(aodiroecfquprueviavekasleavnistt) ililefl Page 42 o f 65 Exygen Research Page 83 of 110 000106 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 ExygenProtocolNumber:P0076 Mn- tenth hutedMntevwoiTij I wrnc*iwwBw MatedofAinlyiitfeito DawmCUlnndmbyL[OFoMfhSi/oMraSactainttAcid(?FOA)inFilhand 12.5 cSauacapiliiasntaqnpdMardMrutfeotmildciisnhoucldbeedtoalttwisdiidrdilurtaedspsaodnore.snAslnyzysdu-nplamu UJ Aq?cmeeOtarie 13J Qfl4fW13Msohmy.iBTihmea4l1i3diimwuppawnnkiqefec(nfc^acjbiutebrloimIhiutP3F6OAemsnifoienn.wihpUarfsraei 132 iMua^teWdMieaet<a3m6uetmne)tnpmmosieBPtP0QlAowutolfMveibl*ogeradltooutihdcs.ntbeLOQ. ifa 13.3 s(JbtUauteeteoirrokatvbaseeoersraoiteaobremaftowohvuieuftecnPdoMpFauOteiattdAeiMV(shipeeidiklehcevdoeeaiartaieo*tntsfeedrKeotmenimnioocwrMtnituMhndiepIO.ifhM-omSOmMpedptuntbeco,letthhhde.eboeeMtmwepweuntbte7lu0lim.*1Ssial0im.Htphokf 134 HmABdo*uqywvttaetTwiebi,tnpmthtnoe^toteerZsshtteteifocwednfiusCedftbhttfeoeatdoeteofteelneeaUutobothntaberaoerutssfatitteiatsaunokdctUaaionlnrUdoonilittdfhiaiOthcubedcojreu.olldhr'wehegetiotehnxcclHuudrttvegede. 133 TheecmUticueoeflkiart (X) te csUbmcurvetravate west he kafif$a9t2el(aH*Jtt&eMffJm).uri1/beiltafttaroratcloems4ujultaet ftnidOinouuntoidrtectphen*tiolenn,itaen,dihtSene 133 RatetemdliaanlatintfaaeatebeertwanartaesftUonfdnoodafteanidboruaimdhpeirtrmausslejwndo.t doli moreUni sie4s%nstlyvtileheiolnmuilBiwsolyIhik*]sentom.uisftrheetaranadnoetilylimzeed.dillincied]diUni wihin M4 CeknalsUeos 14.1 Uhme*e*ealdhneetaefijBbpeoyawfkataaMrgeeeieq}eUuLa*Jysnmb*athrtaocfaaafnkaeontrtptitifIvltecpuinra:(lineoafTrFraOfrsAsinoounndp(uinEnij/amcLn}, PFOAtete(ngtiiL)- lippe 14,2 Unthfolfannfertelo*wccovtttte imouaiofPFOAforo)inn*/mLco ampiWl|ht<f)PSQ A fonte! l'pfO.l * f f f f lA faaad y final v n lu p if ( p L ) X DF1 PP*fttbywfaiehfafaalvotata*wadilute,iffteaeesiry. Page 43 o f 65 Exygen Research ai Page 84 of 1IO 0 0 0 10 , Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Exygen Protocol N um ber P000C764 iw -- * . .. _____ modim vwoma 1 .....analytical a a r u o p | f Ab 1)o1i fordu Detcrnuoitiin ofPoifluoniccMiioic Add (PFOA) io Fuh ind C luubyU V M SM S U .J Fern toplx fintibto with komm 11111111111) of PPOA prior 10extnciioa. um Ab fcJkwiof MftHUoDtoCctlUe tbvpercent toovery. Recov*ry<S)* ftOUl )>< Piyl*foqi)diilpgiitrDt(Di,g)) Exygen Research Page 44 o f6} Page 85 of 110 000108 interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Exygen ProtocolNumber: P0000760 ANALYTICAL METHOD MimiM uster VoeotlM Matte*fAjsljsiitarttOVslftdsstntllsuatOrynLOteMteSrf/lMan8sMluMcAd*(PFOA)I Anfrtteal T stin | Fadlhy; SE0ta5teypClUoMKltenMleiirb,ef>epriuiyttoi AfpmedBy: _____ - j TMtatodUadar,LO-MS,fbtyiaBRttrcb Dite VkaFPrlutaiintoy,QptfitioinBxyj*R#*rtb Exygen Research TttlPlgM! 7 Page 45 <$61 Page 86 of 110 000109 interim Report #23 - Analysis o f Sediment Samples Exygen Study No.: P0000760 Exygen Prolocai Num ber P0000760 M eM N e efcti VflO| 71 _ ANALYTICAL METHOD MthodofAae^fcrihoDrtcroifli>Hofyfluccoet*toicAcid(PPOA)ipVifsiiiiori byLC/MS/MS 1.0 Scope nSnoyieeMmbbnyeeHdebitafjfhltDbP**elrafoaprCo)o/yMMSUl/MqfatdrJdA)iCnbvrtofotsr*ietornga.rd^fqcyuecsotitegtiita*ot/eprfluaornvU>omcUMM)iiue 2.0 Sl* 1 AlwiprtNcveMfcJtaittKyfisetiMi. 2.X CpteicmiotiiaIho**.ippnpiwM9D8biforahurilin^iaychwpicel6x propernTfty 10 Beguine 11 Aiferat20gcftMtuopifarpvirertan. 32 Sfeonftdtrctniinplpettentmkrmimboo,eoplirdSameb4tfi.ipbrrabomduu--cmpflearcettetlouteniffoegttoretuaecndrdrpeyrnkeiucstgeiohlectP.lifeeMicceeieeifhe*wibHeeofomnOrgcpmuiezihubjeanetcmnilatpdimnlirteaiiiienirotnef 5-5 pSmnagfpeapaeiate..eoBtctfaaprceeduieewillbeipeeiedlatlaznpUogplmJbrih <0 IraggiforadSmdrdi 4A444...34312 SMWACtcedtkeerhtsetiomwge-(o1H)Wl(3-6HH00i--P-4HC10L0J0^C0'inLdmgCaerwaegdhrfouy)-f-aJRleeaifgreanitiggrraaddia 44.7fi nuopric*iile(dleO-ILO&ONiHaMi-Rbe)-iIgUBfifglgKraddgendi AA44..I58Ol) DTL1Aot-aOfMieerredtarpAa*Wo~rileRo-HMMePoifidtrlaoC-oifc^gUaaranded*#p-i'iRDetepg^etoigrido 44112) hAeimiluneomdiometAtaotitilAlie-dA-CS.Siy.nHse*iAnml*ieGitrado 50 Initrumeniendfeuiemini 5.1 Abheignhi!peqeuriibprpaewdewettlhliquviednec)h)fMvUotluomgreepliybeccatopracbqlepepbflepeufmirpyeincgtinugp3i*o202 yLeonNctodteeUmbraMepecfeeme'ir{LC/)d$/MS> 55..2) AAMdegvjqisitetllebacloalnlecectnwabdleuci ffomrptfrbaifgciact0e.p0s0i0ic0a)ugt.dqiuaiUstio- Pip]r' Page 46 qfS Exygen Research Page 87 of 110 O O O llO Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 ExygenProtocolNumber:POQO760 foAMUlHRt cfcfetlwdofAnelyrieforttm ANALYTICALMETHOD DatarmanititttofParflionMctaMlcAcid{PFA)inVegetation 55.45 K13f5tunyi-ivpefK-eiML^>r.edfluks. U5S53...7J910 3150amLLitiiptpoOtclfMeforppoellyypprcoeppyjlteenMeoegannrtiiffaigifes tlUi. D12iip5o-maaLbU)*>wPBjriiueupriopwi-m(ScOivttbOCbouLttl.MK.XMOftiU. 3mLlMrHPLCm}ldL 553S5...J1.IfIS342t SAleipteNpiipblitwpi<ji1it0o{iM<0C>iLid10.100pL),withdilpoaeblelip. WCOTraitmettVurilM^ltat(eyaimqfatTefllc,e)m(St.fuvpHlonoiflgat$n0om.NL0p3o7h17'p7f>Wtawtub*n1000qm. 0.0 Qrowmr^McSpNQ 54 (AJYnaNl:yBtic5a0lJC-0o5lu2m13n:F)lutphaaeR?(K*yitwScientific),3.1nunx50ms,5m 44.J2 TMaontpilenPtmfaa*e;e3(A0")C: 2mMAmmoftiuniAeateteinWater 4<L43 MGfoeebdilekaPlfhxaoagan(Bm)r: Methanol JjggJgglgj) 22o0120.o..0005 &<6221A5555 FlowRaw 3377355555 00000.....33333 M 44..70 RlQ^auneae7a1Htmiteatew;Va~cd2u3Fmjaaei:iku1Ae5n|iaL.~(eceoiiebtaeiui>dcmi&Kda4nlo}ecBaloiberfalatiochnuc5u0rvi*e,L>. TcphteimItfoe*fftteOQHwFOLrCiootyratenmin.tendeda*agoideandmaybehangedinorderto 7.0 MS/MSSyMm 7.1 MIToOdAat.BedwprayNegativ*MRMmode,monitoring413-* 169mfrfor Papa)a t' Page 47 o f 6$ Exygen Research ' I ! < Page 88 of 110 O O O lll Interim Report #23 - A nalysis o f Sediment Samples Exygen Study No.: P0000760 Exygen Protocol N um ber P0000750 MMtodftuiwrVPWI71* analyticalmethod MittadofAulyiforIhDetnnmilunofPfrfiuorooottoOfeMid(TTQA)inVmttation ....................... by LC/MS/MS TDbptknbuottl*boOMMSttMtSontyiniumfo.undedui guidein)rneybchangedinorderio 1.1 Mofatiopfaase 11.1 2nMMBCNifunaGMMiwittrJepnpendbyadding(MMgof 92 BxWkSohitfcm 12.1 2c%idMi*eoIOtbidonaLridotfnirtuktniuoo).olk pnpesacibydueotvingZ gofaieotbic 122 t3f0%dinDMitMhyfcl&tfdlfUaMblpbrontettaloio*iofimtpfiovcoata*cinleparfv1p0MmLbwyittahinlogiuiaegee3.mL Alternavotatamtybpnpamd. 9.0 StudiedPhpanben 9.1 SludufSIwkfftelifintkMSoliKMa 9.U JoPMfrcafpnomauloyIMtlicUcatclfe1ua3toa3fcrtdtfLio(LGDoPtfBk*W1b0ot0tfipt.rgrptauLrity)ffuFdOdAilubfycwkd-KjhWingmlL^rnihg 9.1.2 9.1.3 tAmAm1n1L0L112..o1510f2Vfptu5bOthgeLmefUrLa1L1EL.0L0V0fOipomEpgPritbjOffiEafeoiccinnbLaLiOs*hloo.tecttkDuo.ltnsieootoioelhuAUttoiolfao*nnfoicndfafPPulFvPioOOvlueAAmluieiemppo*wrfeoy1pfm0a10rd0aw0ibitbwyhyibtmhbrinehtgoiMigniwngsi1in0l . 9.1.4 9.1J TmafAIlDepbae4p*tm*t4MraathmLletjoiinefiotidttanaafaLenfl*tdify1*oef24ofi30tp*riC.tfnr1iiefMitcpuij,kaaUddgrtaiofmmat*inoEiLnico.osbsluLuoobutltuttiiieiMctioe.f*onarftcFao*FtmoObn2oimAnrdoiu*nnvpodrpelueipnmraioreodr**ooibhffyg61b0rmr0>inotpgwnirtnihthgsi 9.2 sondartiCitibntioeSolution 92.1 pLCfthMeS1A.0Cj9itfnntiLbnfatutiOncsatetinMdasrodliuatitoenp.rependiomethanolviadilution Pipio' P a g e 4 8 q fM xygen Research Page 89 of 110 000112 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P000076Q Exygen Protocol Nurobii: P0000760 Exyyw MrtmlNumberVtt]7t4 I- aiw-yhUlm iim d Mcao4afAjlr'ty.ilfaIlioPa'nnatbiayaLCofMPgSfMlooSrtocUootAnd(TTOA)ioVc(tMcon 922 UftUowiaghlyplwJexampteadditionaloocsomiwimaybe facCMffbummrG0t0i111f0a.hi.144Le$tSansMttiooatUi lM $ V(m$e.t0oi) 21w111,J0000 Di(lmu11i1I>1tt0000t0ooeL00090)d Or(On0M0W0t0P0Qio.0m00f0OOtQWn2n$2Ii$tI$oUcihea 92 i 93.1 aSAtk2lnt"--CatlioXl tPivlCfot,hn1a9tntleaurdtifxdftiuPmfeuffdrlxiiiu,inrtliiZtxSj-iimicLfiLlaUaPtfEbinduirmpwy-abweuptrhepbaordaiies 19.9 8eh9rtUp Itti 10.2 JaaibwUndotqntoUmiqfbicrtourrtlcariMMlrfhctofaytmpsudl(sjflalarnbrtpudclbcModeenltmprtoxiaadlttnn^ictaiJiktoboetrowdt)nianCt(otoiyvpvapyrwkmodajeieiDyrgaltpyi,cjoi2taio0toacodortnnnttnedralirge)ictmootovuteiutriwyfiintifldbcIlrubUedeotttpbaetaiitcccliehfekn.di 11.4 SmpteBratti 12.L 11.2 AiWfcdrfidglifb3y0uSnw$iLotodfKefedfi,'uarantapicitreui)*inalinittidoiohidaikntoai*o5w0jitewmU)rL-ivptoaclytipofnopshyakkoe*rfcteuc-u1r3iAjmgcintuutbae.i 11J Cfeermotitoffitn|cnattwta*S.OmLpoJyprcpyltt*tubcacontaintagmnpletu-2000ipm 1111.14 ttdFPuimruybd.koc3aimh0,*damn2mdL0no1ibdfLtgKlioLoSaCtPMd-ENmulHotStlbt*?eo.S.CtDiunobUaradcaiiatmiOfidtWBai alIlhiwueecdaoltahbleimf2itr.niglaDpfwolwoMnirtiioboktai2l*,pJ0o2l*mdwgfLiftuihlnitoicm.ogalmathl,iun2olagl. Ll.fi 11.2 tDdSfbofiiJait&iymokmemnrcbitftiyytdIttroiblUirn&yndybmtqkiitrtl1fr*'Mi2ft*log$uiLnokiactmr.oatwiL(iCfotiiiinotnIpiDoofaJtte*iitnuteMicatnmiiltnuueaiapdo)teS.tbdutuatPDoitfrn'riBoyu.inatRtcmfoadoeiimefafbie1any3ttt5iiocftiiimtm0taouLminlkitigpUwniertddttou>oeraicltyediItowbpatnemh&eorr-eeosat$tbou0spo%vee,f PageJeP P & 4 9 o f6 S Exygen Research Page 90 of 110 oooH 3 Interim Report #23 - Analysis o f Sediment Samples Exygen Study No.: P0000760 Exygen Protocol Number: P000C76 >mw> MrtbedNtiAnVOMI'>4 AXALWICALMETHOD____________ byLC/MMS Afiid(fFOA)uiM w fa o d o fA ia iy z k fafaf D e iro ifliu tto fN rfle tg u cc laBOk Vegelaiion U1IM1.J9O CIAhedaadlc3r0ifimffcLtehfaeeegLoitaityrfaiklrtfo~frtlbOifeynkpMnliektiuaabfefcoow*aSiNoONttmkiBiLkiiuo*anttesrmihfaaptkiaeetrua.bl-e2.0pm 11.12 11.11 MuDrmtpnraaeuaittpieAcaa,irpeah1at1op.e8oddlmoftosutagokbttIeModEIssaeogmaAtaPb.tEatcowtutrnti.tboCoalliueaeottthtraemhutbeiolienitIihoel 411.11 A 1-4 t o p i r f toctanol I k t n e t in l k p w ahapcd flask and 11.14 mMyikeociihktiflazl^luveoeldinpeie,b*y*isddniinkgf 2a oteiLk ioyfw2%ipeieceoorbr [icaiac4id0*inC)m. ietano!to )U$ *TkeTs""ft"r*?balatrattiloKF"LmC1hv--ie!uaUtgdkpoetb)pipata. 21.16 An4>w rapici tini ekctuejmy LGMS/MS. 12,0 Gtanakpaphy 12.1 kLCn./iMtte8dA1taSznqplwaak.ft ACalibrationstandard.mint3pfaraliAnikdwanndpflealionwkth1e1 ita SratnudtbaeidmiochfPxfFcdOiAneacawKSiapiyotakdiJfagle1i.it leakfiveormoreconcentrationlev*Is II) AnAlin of(notedcalibration*andantemuit baincludedat the 12.4 Uf{cfbbaataotehmnoliogdaaaewunsncltipmclneteieDoeeddidurfBattefnbtcrdtoayafktn&boaatdeobtrmsotrdakdaaakabaanMeotitdtketnhaedxleyneerducvaoareaanvbatrddleyyibcbaeetH5onnot-fttfh1Maafice0arnpvftMtsiqruvroavauemtymataftaarpabpoadndielarekdieaaa.dtuaetatoknuaAnatitx.Mnatc.qhtieeaijnrcEddnLgcxpiiat)antwlr.wJfateatowicaIetnbtrewiMdnedeeniaeiiaitbvghivAtnea*heedmg.ntrriaynaidccprgnadna5leeeo*mre1dcnaf,0uiiptceiroerntvxmeeeufukntsaieepeltarleaoebroedeeeiif 12.5 vSoeaAsnwpulaiceareafiypbiMraarMne.aaitbaouukldrdnomteexmcebeadtwstaonodaairadymMpaonuaijoau. 2AJnyfoirmepquieivalethralit] racaeditmdaidrevpaoeuibooldbefaztbexdilutedandanalyzed. 3,0 AcceptaocaCriteria 13.1 Qodmfv*o1fab3ueUarmobfirekaM(T3baua4cn1tn3ib)aonawpuianppuaanrcakkothoofeuaIsmdapuoogoffdaeatavrtibooontnbasdtioF3aF0iOd0eAa.uaroukfrronm,wahpileairtuet Fa|*ter7 Pag* SOaj 6S Exygen Research Page 91 of 110 000114 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 ExygeoProtocolNumber:PQ000760 Nedtasto?Vooovm AXAUmCAl.MTOOP MefiodofAeelyeiiftrfioDataaaiaHbyittLaCof/rMerSiluAocooctaaacAcid(PPOA)aVtgctnicfl \.2 MafiodMenilitauitno*eeottiP70AstlevalipattarthantheLOQ. ifa mMautat*bNecabtatanneaPdFaOmAiihetfoorlairelipetoaemtaurtathbaaniuP*.$*tprepbotfdie.riam*blankempir 13J U.4 KnfAfiienatotorijrroecvkTnaanreeUitcawoahl;fnotnevifescmb*Ioyupatnbatierne.otfeaIlioxfifncte*fliul*wwfd)oeouhtdenkindsifdt)i-inpeamptbtlaettehaetee*MfiDceipai.eelfkctoMuinulaiiacatJi*ondlneaotluohbtefleietMhbreeobtewcypaetiaMleibfnafrolae7gt0iol-tmh1ne3it0cehu%,urtvhpoee.r UJ BTHboUewAeev*eo*riranfliaenreidloi2tei0)oHmoofulUfnthtbaeirjmo(Rfi)coafuUirnbibraaeHttraobfmjittmaondedvircddusjimdyeuwiiccenodcu.mbiebdeeaxucnlutdebde 13.6 *^tRt0pjri.U9cd9npo2tireriA(iRioi?tuftt2thatDMep.z9ebUftmoa>uwuaN*rItdfMbeertoemnlfthidunaNqedpdieltteoeoerfediieofiunwulUl*ndpifbnliMealltrreaomozuoawteesmliiydencooLtptheedrrereiftttlaimm.ioteerna,dtkhhtehanen . tdHwttda&Bd)tMltUDLirretentiontimedriftexceedtbitlimitwithin ananetyica)ronfiafieaetmub#reanalyzed 14.0 CAkulutae 14.i VbItaaasueamdfiaeeafdotpbloeyawIhkine`agrteboaqu)wuJat.kylnnntufei*eocoftmwlpaditanwlepdrfoiogcruaormvvep:u(lninowfPrFvOjraAnfiocumndpt*inrvrntp*'ttnwLi.i PFOAfind(nftotL) ***ito.jinfamali 143 UBtflefffeiepbfc)f.towtafaquationtocautafiemaudofPFOAfoundInng/mLto W Q A ftxm rfte h l - fP T O A S m M m Ah L I t 9r*l sampleweight<|> nn O f*"fietovbyofikbIbefinalvotatawwdiluted,ifneeeieary. 14.3 fTiebrfaaelbuwpttaaegfoqrtoifiteadtawloifnitkanaowfofiempoanrtcao**fraPoFoOwAryp.riortoextraction.use RecoveryPta- ffoula--frtBfiuodtnrt)oalytcfoundipoamrol(nt/j>3 lQ aifyteadded(ap|) h|t >'? Page S i o f 65 Exygen Research M- . Page 92 of HO (lO U Interim Report #23 - Analysis o f Sediment Samples Exygen Study No.: P0000760 ExygenProtocolNumber:P0000760 AXNaAlMLYNTuImCAbeLrMVWET0H11OISD " HU. kdtfAflilyjtfcrfliDMiimuiwmtdlnUrtvivebrfPLCirfVlieMrago*fr*tMikAiM<FTOa)Im$n*U AcabtkalTntiiitFKilt}'; B3tafxtttaXnCMmoaRUmeiMfefthi.nPDlAkrivI6eB01 ApprcvedBy: CL----iLd TfaeeCht&ticmBloLOayder,LC-M9SxypInlUMweb O..iteitofcM /vj6icqteo/ nFjr5eee-tt5eu,OJpera/tion_*,_fix_yp_a_B_m_ctr_cb Dit Exygen Research ToutPapa: 7 Pa%e 52 o f 65 Page 93 of 110 000116 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 EiygcnProtocolNumberP0000760 XiypeKwwdi VoCJTiS I ... ANALYTICALMETHOD .. MethodefAndy*fartheDMeatmammaiwlUtiewnnbfyPLvOiAMmSo/oMe9tanakAdd(FFOA}inSmal 10 Seofi MSTpbidciaebtrwrydanHartitftifceiaPP>ietMtbcefbwamn{aybatCoMyVeMdLiSfqtrMutfiadJ)edianmncisautitotteBt--|tnadpmlqiayutlmievatyipW.plesdotofpaerftalunodnexmxtwMipaictt 2J> Ssfcty .. : 2.1 AlwgtyiobwvgojbUbwiToryptution, 21 CondllbrqvnipriiiMSOibRftin)uud]iiiiuy(Seiiite4lforprojwifeiy 3.0 finpklbqmnt 3.1 Attea*5| oftailsampfeJprextnctMi. 3.2 3.3 afSs5Soanu|ottadXaubdlpyalJidpmuleepao*ail.ncclwniooa,Apl*awoalotpecpowacbrSnnooWoeao4waitaeplbhlyrleobn.enoepIgdwmfaii*(hpimbwolttaarmwc#imterdmtteUwitb(nbawaeigelrlnefyiwfbsItenaoadnsmvsrdfunyplpwnflehlffeieeccetigiireefl.ibieimnnnidPt.tpaIofikncmFieoboa(tobecu4leaenonUnwatsbssouteeoinrffoorsu6paaprsglviemnkec*dgsvpuublteipnnosla[stan*oiunl*mop!ldfippnmtlil*eioieeiMnidiibnd.soui#csirtf 4.0 ffd?*wdtrdf 4.1 Water-HFLCfriib 44..21 i4Aj PAMAwtoatcfflmuttOaonefrtdoibooUint-t*Ha`A-rHtfoBiCPie*LgACcnd-fdAdra-a.SCdeig^.mKa-4A|idnrticGh(ade 3.0 InatiteMntandequipment 3.1 AsolvheignhtspeqaurfiapupuemdwceibliaquwidietbfclerwvaoaImtomgniyqljiecoatoprabcalepaobflepeufmirjpeinegiinugp5i*n2020 3553....1324 5A1AM05nmdLaMeloyiv,Ltodiiccdbneaispttlptoohaoo*ccsoaoa--tfeeUbi--d*leaptoopdtamdyytpvimprderoadytpnaojrylQffsiorMeirMapcdcewieiuanvktgSrnfifmnprtiitfgeel0igegc.0armt0astu0tbiMob0cen1eis.e$.rnadaqMua^n^tXitation. 555...07 L2D25iqj4L>WocLnJmLblDteKnfBPcL--foCpitveipiwoai*lmt(o5o0tn*lOttOboutLtle>*I.QD-ZOOuL). Exygen Research Page 94 o f 110 000117 Interim Report #23 - Analysis o f Sediment Samples Exygen Study No.: P0000760 ExygeaProtooolNiunber:P0000760 >""* MuMMurtWVWPH I aaa.ytm :a l m k t h o p MrfcajnfAMlyit.oftrllirMiitammiiiainlltjwiKbiifygllCliiM/MuSecMMSaMeAtMffroAibi'SiMH 1 $& DfepoafckpifMM. 1555..11121220 SW'AHPuimMuIc*y*irttelSipzwaitpKMH.m{lkO*VJ^fmcUld)0N.iiLaMCtlH0>9HP)E0iilfc),twridjgibd.bpwbleiipi. 55..1154 CWaritMtriCbfk*MHfBabt>ttkksa: fjfrkf15cnLpotypropylfntub**M3000rpm. 6j0 Ctawkykrftittym m 11 (AWidNy:tI2k5a&lCMoUto2n13a0:P)hio>fcMR>(KytonScitatiiic).2.1oboi i JQmnvV 1132 )TAwattp>mFthwiM*: (WA)C: 2mMAjcauajunAeetemWater 1134 CMonbickdF:fPcaritcjp(Bu):: M a te d - jQvh0(1l...0^00hb) 2220..30 116555 1655 &35S 3755 3753 P/rJawlJtwmiwoi 00O..D31 01.33 16?4 QkjottotrttoitanbVoortaPmtio:k15xpwL(-cdbobmtminUrnntiartti1cdUtanutloehDucu3n0*h.L) 18 RimTom:**23minuta. oItoptaitata*ih#oHedPUICtoaipamin.tanto!Mt juidm i miyU luogo*inoitaio 7,0 U $M 33yw tt 7.) MoteBtatoipayNofrtvoMRMmotatatari#<13-- 369irtfor T fQ K . Topbftaiubtohi*acMonSdMitSeiyrrat.iatadidm pudandtu/ bt dunfpd inorderto Exygen Research ~t -$ * M * * Page 54 o f 65 Page 95 of 110 000118 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 ExygmProtocolNumber:P0000760 ftypalanini_____ taHkriNanfa*VTOOPIJ I NALVUCAtMfTHOD Mi^ofA^fatlwrirTinMllallnrtnMnagiLrrvwarflbyLBCoAIaSmIUEoicdd(FFOAJiil3roll 10 PrqpMttatof&ftfatfttU (.1 MnUblbw AM 2ammmMaanritwbnaacnehtaMtetaocttaOaOtaOlanLwoaffewraUfepr.nwudbyMin*O.LV**of AlternateyritaiMmqtInpnyand. 9.0 itmdirtPrmamte 9.1 StandardStocOTortiflcatiooSolution 9.U o*nfepaanlytatedae*cmkuataotteodtio(ofteonfw-1ta0d0tueprptoutLritoyfPmPdOdAilubtyevtotifh10m0geL10wmithg nadUBOlinal23mLU2F6bottle. >U AmtnaILJlZopStfttmabLlL1tLe0D0rtPipfigEMtabtlioLDManaoa.XpdilottoNtoeofffUFuOlvAoJifutpmttrpoaff1e0d0bwjrithbrminegfitnigw!l 9.1.3 aaAL102o.51fmte{ifl*/.iniQ1L.0fPe|iiBgtifttteaoLattlttu-oaluafoiMlultoioiafoinfpaplvtMoluiampereopf1v0M0lwbyithbrmtAetghainaogli1n0 9.M TbeatockutaIbrtUScrtiuapotationarttobtitcredinarefrigeratorli afaputMntMhhnudaaMlelyo4f*pCepanmdaatimn.atebteforamaximumperiodof6 month 97 StandardCalibrateSoteteea 97.1 9.27 oTLfVDtiwMfOoSl.o1IMwp5inggfraiiLillfkntaittyteilpaeicaaattfalonanadaamrodpiu*ltaew:rnta.pdndpitaiorncdalincoBnMcethnmtraati)ovnisamdialuytioben S9nCd7fouJfuaierB1I1f71tWa,i99S.9ff99nt99iLrttaltniiadrMtaaii.1l eaVfi<bo311111*r.t4400U00tetouattadnRrdh(m1It111rTow9009OU9999ltadto125-mLCLmDOcPFi9MA32991Em.....14n9930panLaiJatiozrnow-moutkboidm 97.4 itA2i*taCvtaotei*Cvo,itpattead*xanid0onMAanl.ntooofKandardamaybepttpartd** Page SS o f 65 Exygen Research Page 96 of 110 000119 Interim Report #23 - Analysis o f Sediment Samples Exygen Study No.: P0000760 Bxygen Protocol Number P0000760 BVPMMM V00OI7U I ANALYTICALMBTHOP ' MltodefAiwljiiuftnlwDMawmifminallnlLltivilelrobfyPLirCflMinSw/oMjSwwtoAeM(PW>A)inSmal 10.0 BllchtaUp 10.1 10J 'wEkemitq*wutlmeinrtkemttbaiaeta*slidfbQercmafabipercltledoeea)aleinumxditpralaicbctetCewd)not(toDtyvurpyeaicdtrniaufwypllyptfaircdo3itc0oaacodoawrnnIatndr)oneil*)pcaoiktmvtoaetirfylnatHeodlub0demecpbkeanctlocierhtwaasndt intoputtitymummplantothisproject ILO SempleBxmedoc 11.1 111UJ AWotHneeiaMdadiiwne^tdortaa,)|jdmst$edpreiolt*epteftcewootnsdltiMpdilnflJgeeaBnmpdinliemtDotimoii!xftaiw9tmkf0eimnpln)al.eLlNvanpoleloclsuelrymitreptftvb#rrtoaaropi~letfyolb1lneal0endumefooteuLrmw.mreig.hiaotoMbwW(efcrriitirifyybue 1L4 TflanppoeiabbleIBmtotLdfUogfethtuebeampleuaingdlipMtbk pipetteinto i l) mL 1111..99 CAdadu9tomflijeLo(hfeletuebtoeo*iiltril*o>1m00d0(tbpantefafor-r**34m0inmuitneut.o*onawrist-actionihiker. 11.7 Docentthefcperubntmtoa 90mLdisposableeentrlftfemb#*rxJadd35 b L ofwart. 1tL1uL.P10 lCLB1b9ehottamrenieddwLdiwtepiwooidntshlhjaptyhmr$eeppmCmlyeiLleomLoaSoefeHPofDPmoaeLdfaeCltttiiffhOutwargMindieModftlturCt(b`Gi(-eS31(bdPfi^fenlcE6acoptlB^v2mileeiUmLctu)g).LnebD.yooDpf2nicaseJcmirotainlaLerdgt)tt.aei1nlou0thosm*r.aLigmmraaedtdhuaarynteed) 11.11 AltolywJeoplmsineelect>o|iCaylC/M8^(S. 12.1 tycctbeaameamountofeachkmrtrdtsaapfeandfortifiedsampleintoto LanOaMlyz&eMdSmspyteeta.oaAaHfanttaetaadaedmust precedeandfolkal 112 mSiaudadb-edenroiufTdPadQiAnaonemlueapljortiHfeKanlgtotto.atlaaelfiveorrosteooticcrttxahcaIcvcb 112 12-. geaaUitiAnnhaemtmjnekemntoicepaclpptrranhleUaeaddodtaertmiu.iooedasmetaeftd.Ao,eeteetaohAtvamumeefUslerepyJaabmlauUo5efl-ilbttlte1gycncr0eattMnuonlaarfutovbtmatSsoieyvnsisfpeedLtl.naeainoidhaoceftnddroa(sadrtaqironrssmdugateituricRttecutsciolubetoMnartubfiotbunneobletucoelotoLuuhwtsifeorleeicntcdaafgLsplmlleiieneblbc/rerdoxeyaanaetwaridnodctedniaettbaihgelielnbtealhitadrwostbaiitanKeeftisngieogntua)ninojnoedfeendccpvmiatenieeormgmadvrnynkat*asnd)ay5i.andrare.db)Irainelafest Pm Page 56 of 65 Exygen Research i %s pagc 97 of 110 000120 Interim Report #23 - Analysis o f Sediment Samples Exygen Study No.: P0000760 fcxygProtoco)Number:PQD00760 rnk HteaJ HinterVQOQJ713 I ANALYTICAL JHETHOP MethodcfAiutalafafaDMetaommmkaxlltilvetarbryPLsOrihMmSw4cuofaAold(PFOA)in Smal sVoWfVtwUBaeretfeihymtmo.r'itandardeceuetmioaminiM*nLytw3.3 (orequivalere) US SeximcepeldemmxidpinndiuiisdxpteouaUittenfeatttwii eteedfurttahnedradrdhfwloapomneiani,a Anyunties to i/u A . ihre 13.0 AcceptanceCriteri* 13.1 oCfh4r1o3Mmto*fum.mItemu4f1i3ebamewitppeenlalieetoanadaupboiotda*rtoondieetF3P6Q9 An uamfeanrwahipleeriilrili 13.2 133 dbIfWmtawinaeidiegrotethtkettvcenedoostobnowIbnttanteaatveiCnefa!meWPidamFslam\mOMin.dAulItnhfi)petailrlklpsecewoovoentawstaotlaminnpnmdwiiemaPiiapntFdeatulOectierelAiefxftlbiimencetpoiriltae)fka0-vxenaaxtlmeatme*dajgeo#ten.tefdftaadtileoc.nxratiedcathcetwane.nepMwtaihnbUle7m0Lli-Om1k3Qt0eit.9im,1Jiph/pie>f . UenGtirHesfaetuoMfeelonpelevealuttaoteedldbbye-nfaheaxntraelcytsetdteAdneytemrmaintreixifpfeil~eeoautniacMtwoQ70it- 17.4 13.3 TAEmHhnruoreyowturmceaTXmledist,aetn(,tltlrnaeitetmunttiopliUmfcXboHeDeBIfflotufxeiccsfditebfoaantduerttdaoo(Rdtftiole)immosfatmtoerfrhatc--eaaclirb)tfteiaetaxxttuikonrafraiitoxdrcatuefxroovifntefaitjsfeeatngboccycodnxueiuUlrdaitioosbdet|tkfeamxcuclHKuudrieyebde.t jttffaeXdPfuattd(diRe1rf2fa&aPif4a5en)w.aetlIefbdeuoiiUbffirctametaioptaek*r*de4swjwuult?sldtOioeifielreuoaantfKdalndytzsedtph.eesnetoliamailin.dihfean I li R*xn4cnteHurtlyi*atnifeaaloinlnieoiaftnaeaonlryeftaisciaslneetimmlul[sxinirbtfeeinmetniondenulytiotmadpe.id*rimfteuxscteneodtsdthrisiftlimmoitrewiifumii 14J UtjeasneeeafdmaoendfbpbfeayafakafaaMrefaau)ttueLaiyioxnfxtfolaecAaiwtlcanudrlaestpeidrfoagsruaarmmra:o(ulinnteoafrPrifjOrcAwibiounndp(inrainMgUtonL> PFOAfami(nfteL) dnlundmeseti.xDP<fareifalor IAIPfa-pn(blefatetebyw1h0ichfa finalvolarnereediluted,ifaeocaiary. Pig*iaf ' Page 5 7 o fS i Exygen Research Page 98 of 110 000121 Interim Report #23 - Analysis of Sediment Samples n. 99 Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 VOM) I A H M Y nC M , METHOD M ahalefAadyibfteiluDeunelmilcofportoonocUnckArid(PPOA)laSmal . - U m nulUnrbylCM SrM S 14.1 Fee nmol-- M ilM wlUi tnowu m o u rn o f PPOA prior u unatriui, w IteM kwiogoquIlMM colnlmltwpeKMlnamry. - ,Rseovwy [lotelpuly>BumdQiaM.)- opolytrfaundincoarrol(ngrinL)) toriyM-kM(a^iiiL) * 14.1 Um thtfeOriwiagequraioorov--n h e w --uvtofPFOA foumlm ripftnLto >*<*), puh* ennuifnnti-mmAe--<miIp--la'mwLrigiilne(gn)tivoIqmrmin r*- Exygen Research , ;*- V x T ' * Page 58 o f 65 Page 99 of 110 000122 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 ANALYTICAL METHOD M edndNm ter Vexol? Mrked g/Aeetnli b ribe PtferaBeaee ef Perilpuniecleeele AcM(PPOA)li sm u M em el ( e r r a t? LCMB/M8 AubtiulIcethjJ'eeUity . BeygetKeeeeitk J3c0e3c8rCReeilitgarec,faFDAr1iv3eMI AppiovtfBy: PTeeeetlmCioce!eLNcteyrier,LCc4U,jExOyftri1Reectrch DattleliA/m ' JVfoiceftaPnaaRidflynieOpmtiflM,ExygtoRewrek /'J d r Dete Exygen Research TetetPejot-' 7 Pttge 59 o f61 Page 100 of 110 000123 Interim Report #23 - Analysis of Sediment Samples n . 101 Exygen Study No.: P0000760 Bxygen Protocol M usber: P000076D b^nkiM M ih MattedM nterYnome I *WALVTICAXMTHOP M ated ofAmlym forth* r>iHrtrrmrrttir>nefTeritowooctooic Acid (PPOA) u> Smell Maanmil 9iura by LC/MS/MS 1.0 Scop iV* method to be employed for Um iwbticn mid quantitation of peffloorooctuoic aSc^idettbMyMJn|cbDPemtefcotonrM(LiOnMSL^iqMuiSd.)iCnhbcrmeImInoepapuhMy]aceonuimpl.ed to b tondem M ui 2.0 Siftty 22..12 ACtoweacytaditltbeiipmpiroi plitfiKttleibMwSiDteStybpeifeotrieciwro.dlmtMychemicalIbrpropertafeiy pwwii-- . 3.0 Sropl* Requirement 33..12 3.3 5ANtaititmobalapealsercatetpIenobuSLpacetRtoootkefmaeteailepoairwiaaafmeciadortdonotbencatfotoemwrdepixflleoltrbtreaealcyrtgetihoucanmcwtfUatoadamapinolemath.teaHmaomwpeapvnleionrgnfbptoelamfnonrefoauraan(ehw.ien paejeet 40 ArepsiarodSteidarfc 4412 MWeatfbemr-wHPI-LHCPjntCttfreed 444AJJ APAaaeuroaftetioawocnofomoUcAtewc-HentoPaAtiCeo-jiArde^.dCSe.i5u,mJUtoeMpdiortleGbrid 3.0 InctrawatandEquipment 3.1 Ahi$hperfemncaliquidcftrecutopiph oipabtoof puafsngupto2 tetventiquippedwith*friablereturn*Iqfoctorc?aMeofvertrag3*200 33..32 S555.A473 AA50ndnaeeloLyvotiidcMiapotlootbecadalUaMtontceoaetptaomonalwydppetdeematootMpeoyfkofuroreteaScdpeineagtoicrttiotferUtjt0wrg.a0tite0twu0rb0o(et1Csa-n/M.dq&uMan$tit)a.tion. DtI2^i^i5uohIsfbdaiLiftaoLboDscdEcObfiTttomppipiOoeltwfefcC-iHaSpQQyul-IilhDObiOcottotLfi,Uto1U.00ta*e2o0f*o0e.). 55.J0 2DmisLadbeelerHpPipLeCttevsW. hit 55S51.1.1U1032 SAVWPuoaBtrotairpnenfoeScu.eSnpteF(da1el0taV0n*ci1feo0l0t0L0ccp(Llgro|dtC10ll-180P0EueUmitwridiptheed.isposabletipi. Page 60of6$ Exygen Research < t * 5t i Page 101 o f 110 000124 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 ExyjenProtocolNumberP000076 Muto*Nur*VQOOirib I A>i,U.ynCAtMSTUOD MettpdafAadyuiefrtimPMatemnmiHeolSneiorunmbftayLftmC'MoMi/MteSrerieArid(PFOA)in5mn 55..1154 CWftiUlinrifaatfoocifigcaibiMJar.afiptiin^ 13mLpolypropylenetubatl3QQQrpm. 6,0 ChmeuejriphleSyriern 6.1 (APM/Nly:8lie23e0lC5*o0l5u2m1n3:0F)fciopbee*RP(Xeyeton*Scientific),2.1mota50f*n.5H 66M72 6.5 MMQTmsoodbmbitefoinufPetBfPcfBaltteottcaofra(i3WAmd)*;:C: 3MmaMtiuAMmlmoniumAcantiinWater ' TW220S120f.t.0.09a50laV S655A 22555 s337355a 7355 FfaloiL0w0000/.....m33333JUintl 66..021 QRInuujeniecTttiiiUonnttoiV:ao^:lu2Pm3eaenk:imAl5ina|ictL-.e(etXmbciJjrtenewttowdiolibBrne>ttcoh!*ciu5r0veM,L). ThebovtcaotiiceiKaretnindadift(indBandmaybechangedinontoto <VtmB(beHPLijotacL 70 MS/MSSyuero 7.1 MPFodeA;.BbctwjaayNejtfiviMRMUMde,aioaitBriji}4l3-*Jin*fai - oTphteimisbeovtheeoMoSndMtSenyieemnm.intendedm i jude endmey he ctamjed io orte to 60 PnpestiofSotationi t.i MobilePheat 6.1.1 w2waiaMonummatoonrtiuemlotoelOieOtimoLw.oeTmwaieterp.ropiredbyaddiaf0154gof AtaroitevohutMmaybepiepend. repli" Pagi 6 of65 Exygen Research Page 102 of 110 000125 Interim Report #23 - Analysis of Sediment Samples p. 103 Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 SwmRamftb MtoodMwibNVWOmA 1 ANALYTICALMXTHQD 4atiWdefAnd)k(brDMM*mSmMdSSccr0a9mrPbyciLhCM/MraS(/>Me$ttMicAcid(PFOA)aSmal 10 StandardProparato* 0.1 9ta.1rJdanRtSetpoed/rFtia*r*tieflckaitchmitioSnfrleotfi-o*n106p/mLofPfOAby IDmy , ojmfmanhlymtoriaaJ1m25la^*1l.(oLUcPifrEeubdetftoir.purity)anidiluteto100 rotwith 9.1.2 Aminf1Lr.0firpftbfmmlILlWLDJcPjuiEft'iifitiLfalitottamoohtoliJnuitfloan*tffimPFJOvAohhtmprteopfatroeodwbyithbnmn(ihintnpo1l 9.1J . 9.1.4 aATffmiroph1LnOpe2srei5ioltthft*paBiedr|mLetdtaitLaaeULOnt1e>dyT*fpobH4rpcR"pbtCtrioioekBLaptiiKenaidt.roilaouomtaatepo*u.oil*dlbulberliiobmnOfaaibferPin]aFvjtOnooalAubxmeibxnietpeuorrr*ofap1pei0rei0*nrdiwobdirlyddoHbmlrdgiancingmraeinnoloognrthIiiOnt* 9.2 StatoriCililntiMSoluto 9.2.1 9.2-2 LoTCbfffrtft*fBc0l/Mo.1wSpfogMgtalkifalfrforattryotipfaictaaatta)tinwedximintloupmtiboo:np".eI^ymmodIinmmsLecthimaniotilmviiidmiluiytiobne Cooombciwa fVmtUkiton Aohlktt fortnD 100 100 ICO 54 24 14 Vota* CnOO JjO 24 14 10 10 10 Diluted te <TMw 100 100 1 100 100 100 Find CoftMAfrvcioa (u 'm U S4 24 10 0.5 0.2 0.1 Aitb2*oCut*0t*oC!u,aOepetooikl!lefiottwoetin*t.racionioftutorfemiybcpreparedu 10.0 BatchS*iUp 10.1 efa bods of staple exneetad (tjpieilly 20 or hm> rouft toclLtd* d lean CM reread torero) and two imhiiim) coom b fortified at known 10.2 eRocqDiMMTeronetBoDotiil(olarbScaoidntaronldslpafbco*r)afatorvyerdiufypplieroacueaduarned) rsepcikoevserwy filolrbt*hesptaeechifie.d lo ih t quetityiswiDe* pUa for this projttT. Ptfriaeir Page 62 o f 65 Exygen Research <bt . {, Page 103 o f l 10 0 l2 Interim Report #23 - Analysis o f Sediment Samples 11.101 Exygen Study No.: P0000760 ExygenProtocolNumber.P00007W MmMNufkWTV000PI* I AWAIYTICAL M tTHO P .... j MwtwdtAmljettfcnbe'ItumlnHDffcrfluoroocieaoicAeaffFOA)inSmilI M auul Scnui V} LCMS/MS tt.O SampleSxoicob 11.1 awMnhnyewbeedmeid1a,nnmuLprebrdfewdaKapdipaanlntddininditotrnt5wh0eemml).alppNloeoityalpaiswhanyvyaalilqltaetbrtnoleamtfet*rcifmtot|leu.mmnbaofifmetuiimfy 112 Addvmrtoilsample&rillrulvolumeof2Pml. Ceptigttly ' I11.4 11.5 AVdTitdoapdcnet5aeKfamnfhiLfrt1-oc1mfeejdlcnamrifofluatiftnyeIi.klrtuilBbeea.nadn4pltealmuiffianf-*-a20dmkpineMalufeleonpeipwemttet*einettoionash1e5ltemr.L 11L1.?4 11IU1L.P1't 11.11 bDCCIBfItwSAtalLaiopwnmucdtowad*MLkltlfybweipwifottleodiMaicihtith.*tbayaeUptyarMa-t.imo2JeoqpmmpmpC)dt4awLknbLcuoixnDr5aneefwHPUinfel6^ifmJta3Circrufier0caatwrte0cihrlbat0atir4lrsinrdotpaf5orfqiM(mlC0t.'uj-n2bnufyC(oda1LSLror(-e(^HCld5bepi/sBMuc4Bip'ttcSoajv3enerb/otMaacrLmlilhude>S)lsJamy,b-.D.yoeeoD"pfna2aieecitfMiracrtiilhMLioord|Sgteacoi)ntlQuulutbometmc.Lanninnndmopatuhddtadityen3dp5i 12.9 Chrotnatopijhy 12.1 lLaCje/MctStb/MaaSimayiuMron.MAloofdiabcnbtfUilamadastradn,dsaarmdptlnewenrtdp&arctJefdl<dasoadmpfloelionwtoUaKl I2J 12J 124 12.5 gIxeAtuiMcUSirwSnbdMaximmratjMantefctanhaaatmewceecpelffnkjyetbtapernrbekSaetdiilepedetadwenrtcdeau*rdo.daaxitaeirtarIaeiieynlfue.inttebradAtfi,dtbql'prsaMofaswacsPorCtimalricaastbactMiadniafosKmee^enaadnnbyMp.samtirnbataialprbe5tdhttoafmak*Mirfe)o1oMunat*c0aireelmtcBfauidettpeetrsiaetbvadilfnaStvttlaayenityeol-ailnabptottt,tadiniedlesoeofaedobaUenaelxrfafnldrridiacgf(yrdsaeabtrondqoaneteinwttour*d.aftssrafiotematnaiaeimnoottitettaoanlibetatiaunWfoeaomcbatdibBrnlteotuaodeofntndfotmwhillii.gvunflerielnrectMeaedlrgaautLoadaaoptdpeireiaa1nbbemoetmaAnruydp?rLedtoaaxmeywWtmndinrdcoedXcl.aa(tUwbei)ielfkieIho3bnataAat,oftrdwJ3ntblrnaeiynaneefoytt(igng/teidogenduqnstdinvrounrjcie.medinecdcfiqruipvisnaipatureeotgflvnimieiearJvneyakcalaaesl)Mylneaiev.nthneonmbOIarJlntaeei*)tti tlfiJCf* djPage 65 o f Exygen Research Page 104 of 110 000127 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 SxyRbo Protocol Number: P0000160 wb - ________ tpttoJ Hubw V00017H I - - ANALYTICALMETHOD 3 * ^ w 5 w i ^ S 3 5 5 n i m w S l5 i n i h d t o w ^ S JtoMfffQA) in Smfclt ~ .................. Munmd Saum >y IjC/MS/m s 13.0 M tftumeritarla ' v 11.33...21...rCobtal^fifna4g1knh3mtcir*oMmjosqpta.i0iiBT6h9tnTnemuO4ai1ntiA1a)bonetmpe*rwleiapviawpaatauetmabaknpPIohwPafOtetlwaoAdwratepuUoofIahrfwoetavir1btioa0oynrfitMata/foitPfs3LgFt6d,O9Q*-Aepm>uuihcaR*nuLn,mQ*wwQtipt.bicaEtrae[rrniteliit! . . . 13.3 134 .8dw1en3ntpai0miyrg%iiiscehratmmiattbwRibdiupwfnfmHcUfemviebvaiesatbaaio/ekpttbtbatUemieu.voelsaIhfaftfiechoiddnltoMnaaodafntdihUuaadodsobtltfbdoywgfetrroi*titwfmaaumpabiairferbtketeiteataahiecll)wlranhnolueatlciaflnreiobbluiAl#tlidiodq/n!*arpo-yatiahumdoaxUaeiUfmetmhMvrici*ubMwitycrripdpabwni.tektib-bebelerfaolmi|iiiwlouitmhcaftljtafiirccnlHiiutn,7ur?vit(e0eMjea.-30Sof . Howewr,thMalnueteofeatifanttateinb fu* couidbeeieJkdod isi TaBp0hUpe4Mr9one2peorimai(Jlatic*MariCet*aBd.f9a2|cSe0mc)*.daPctfIiffutbeeutttoftb(atRakne])tMnnfceBioruberawedufjotmharitiaieJbDekuUttorcueudamr*vtafeitndaicfotetphdaec*teaetidmlm.mitiuu.stithtchbi>ec 13.6 mRitm4aai%adoraintJwwlyanitiohodtirni)mtmbenianIbbiraaaatovwljiitekkhatauMlteiuttitcoa.idiumaItfrdbpelieiinacliiatxaian)haealuymtilnedpttdebia-edrriaftajeinuitacWleyzemcddi.dtirrii*fllummoiiawithhainn 14.0 Cafcelaitons 14.1 U|bWaMaaMtdfMtob.JfopblaMyatvhkbintMae)vtaudLtjUy|na*iPtfstcsoaftawlcteuadrlaeatpBrdr(ohceguarrvama:e(lunitlaetfPrnFOpwAfioiwundp(ithncim\|AiintrLe.i . antrta 'f- fpgfrrHtop* DAPliqutMelobcrtbayr*f2tfc0idilhafinalvohunewudihusd,ifosccMiry. 14J dForftMHtot^wdainagieoqctuifaitaiodnwobalakutoilwMncisbsaupieBrciteimofrePePoOvAo^p.rierioa x im tio n , u k Recovery(K) FMaliUlWteBd(BglM3ana^lfcBBdpeeBiaol<B0^1.)} _____ .Mlv>Mwt(aWj________ l __F_#f4_o* ' Page 64 o f6$ Exygen Research '! , Page 105 of 110 000128 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Exygc Protocol Number: P0000750 B iffttU mn t j M cbodN ui^ti VM0I7I1 WAimCAiMITBOD ....... MwnraiSerumbyLDM5/MSMethod rfA ia ly iU fc c S Ito te in a M lia B o /frB rlW ra c c ta a o k AndTPFt^ATm Small 14J UM*t&l3awfa| aquationto convert Ihflmoatf of FFQA found in n|/mL to PI*. w in * 'rr*.^ '"*' *--J ' f --1--fr-- 'rrt H t t m p b toIuitw (mlO Exygen Research r |> T fr Page 65 o f 65 Page 106 of 110 000129 Interim Report #23 - Analysis o f Sediment Samples n va Exygen Study No. : P0000760 n RESEARCH 3058 Research Drive Phone: 014*272-1039 State College, PA 16801 Tax: 814-231-1580 PROTOCOL AMENDMENT Amendment Number 1 Effective Date: 01/19/05 Exygen Study Number: P0001131 Cileni Study N u m b er. Pegel of 1 Npffl DESCRIPTION OF AMENDED SECTION 1) Analytical Procedure Summary V 0001780;Section . 1 2 ) Verification o f Analytical Procedure AMENDEDTO 1) Add to Section 8 .): Section 9.1.8, Alternale weight of standards may be used to prepare alternale 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 sb e available for extraction and/or to cover analyte concentrations expected In the samples. RATIONALE 1) H g h er concentrations of standards need to be prepared In order to spike the sample bottiee at higher levels. 2) The sample sure avaleW e for small mammal 8ver and serum was sm aler than expected. SpHdng at the pre-dstermlned levels In the protocol puts the spliced concentration lower than the detection Until Also, the analyte levels in the ground water samples are expected to greatly exceed the predetermined spiking levels listed in the protocol. W hen ths levels In the samples greatly exceed the pfldng levels, an accurate neawery value cannot be calculated for the Q C sample. Higher spiking levels In the bottle* will cover the analyte concentrations expected In ihe w ater samples. _ IMPACT ON STUPY T heL O Q is 100 ng/g far a 0.1 g sample o f small mammal lhm ran d ts1000n g /tn L fara 0.01 mL sample of small mammal serum. Higher levels of spiking for Ihs water samples will ensure that more Q C recovery data can be used. LIBRARY ID: W0001226-8 t Dm Jfa ft* J * /* Exvaen QAU Review 1 .11 o r ili I f ADMINISTRATIVE FORM Exygen Research Page 107 of 110 000130 Interim Report #23 - Analysis o f Sediment Samples Ti. <100 Exygen Study No.: P0000760 3058 Research Drive Phone: 814-272-1039 State Coltese, PA 16KM Fax: 814-231-1580 Amendment Number Effective Date: Exygen Study Number PROTOCOL AMENDMENT 2 03/07/05 P0001131 Client Study Number: ^ ia1 Nona DESCRIPTION OF AMENDED SECTION 1) Report, page 11 o f 65 2 ) Teel Matriels, page 8 o f 85: P F 0 8 transition monitored 489 -> 99. AMENDED TO 1) Instead o fo r final report, interim port will be Issued. 2) PFOSbansWon monitored may also be 4 9 9 -> 80. RATIONALE 1} Due to Iheexoesaivs sizes o f ihe data sate, Interim reports w ll be Issued to allow the ctont to receive data in a timelier manan '' >*<*' l* r W 2 ) The API 4000 LC/MS/MS systems detect toe 499 > 80 PFOS transition with greater sensitivity than the 499 -> 99 transition. IMPACT O N STUDY 1) The d e n t wtil be able to receive and review the data more qutokfy. 2} The 499 -> 80 transition can be detected with greater sensitivity; therefore, giving better chromatography. LBRARV IO: V000123M - Exygen QAU Rsvtew. L u a h s f a ADUtNISTRTiyE FORM Exygen Research Page 108 o f 110 000131 Interim Report #23 - Analysis of Sediment Samples p. 109 Exygen Study No.: P0000760 t d m * RESEARCH 3058 Research Drive Phone: 814-272-1039 State College, PA 16801 Fax: 814-231 *1580 DEVIATION FORM _______________________ . -________________________________________ Pag 1 of 2 Sanaa!; X Project Specific Deviation ____ Facility Deviation Date of Occurrence: 04/26/06 Exygen Prefect # : P78WP1131 Deviation # : 5 Client P ro je c t# : _______ NA_______ R eference#: 06-076 Regulatory D river: X _____ _____ GMP GLP Other None Sam ple DeecrloUon: Deviation Type: (Inctude VH fa r m ethods and SOPs) X Protocol _____ Method _____ SO P W : NA Notebook reference: NA L o g in # : L0DP8191 C ontainer#: _______ NA_______ L o t# : __________ NA Sum m ary Of Deviation: The three sediment samples In L8191 (C0172892 - C 0172894) were originally extracted using the sediment method V 0001762: Poor recoveries were obtained for PFOS, PFO A and t5C PFOA. Because of this, the study sponsor requested the u3e of an alternative extraction for these compounds, as follows: D irect Infection M ethod; Before the samples were weighed for the extraction, they were mixed thoroughly by vigorously shaking the container. A one-gram portion of sediment was weighed into a 15-miffifrter centrifuge tube for the extraction. Ten milliliters o f 1% acetic acid in methanol was added to each sample. The samples were then shaken by hand, vortexed, and sonicated for thirty minutes. The samples were then centrifuged for -1 0 minutes at -3 0 0 0 rpm. Each sample was analyzed by LC/M SiM S elactroapray. Using this method acceptable data was obtained for PFO S. but the recoveries for PFOA and 15C PFOA w ere s ill poor. Another alternative method was then used for PFOA and ,3C PFOA, as follows; A lternative SPE M ethod: The samples that w ere prepared In 1% acetic acid tor the direct injection method w ere used for this extraction. Five milliliters of Bach sample was aliquoled into a 50-rnL polypropylene centrifuge tube and the volume was taken to 40 mL with water. The samples were then ' centrifuged for -1 0 minutes at -3 0 0 0 rpm. The supernatant was then loaded onto a C ig SPE cartridge conditioned with ID mL of methanol and 5 mL of water. The eluate was discarded. Approximately five mlSillters o f methanol was added to the cartridge. Five milliliters o f eluate was collected into a graduated 15 mL polypropylene centrifuge tube. Each sample was analyzed by LC/MS/MS electrospray. Cause: ____ Preparation____ A nalysis_____ Instrum ent____ Client Request X Other Im tii& b More usable data was obtained. : LIBRARY ID: V0001B40-6 fcxygen Research ADMINISTRATIVE FORM Rage lUy ol liU 0001.12 Interim Report #23 - Analysis of Sediment Samples Exygcn Study No.: P0000760 CHEH EHSR 236 1B 651 733 1955 .. B M l-M O I fltiilw FraMUTTW Ltm O B 05/01 `06 13:66 N0.106 03/03 + i-iM r m nn wh JUtfeKWChlMv P te w rli+ 1 S titt esU*t, PA1W1 RMtlA-t SEARCH O fY ifcTm rW ; -i , , unMwiRvasaiUM Exygen Research .* s v r AOBWTIW^IS Page 110 of 110 000133