Document zQ51RM6XRnMXexNRnN3J1dw76

-n '!-/ f~~ ~a6_ 37,9-7 INTERIM REPORT #23 Aaaivsis of Sediment SamAles 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 DATA REOUIREMENTS EPA TSCA Good Laboratory Practice Standards 40 CFR 792 STUDY DIRECTOR Jaisimha Kesari P.E., DEE Weston Solutions, Inc. 1400 Weston Way West Chester, PA 19380 Phone: 610-701-3761 INTERIM REPORT COMPLETION DATE May 2, 2006 PERFORMING LABORATORY Exygen Research 3058 Research Drive State College, PA 16801 Phone: 814-272-1039 ,~ _STUDY SPONSOR 3M Company 3M Building 0236-01-0-10 St. Paul, MI+i 55144 Phone: 651-733-6374 r".) PROJECT Protocol Number: P0000760 Exygen Study Number: P0000760 Total Pages: 110 BEST COPY AVAILABLE bo;wlc Documenf] 000024 Interim Report #23 -Analysis of Sediment Samples Exygen Study No . : PQD0076D GOD LABORATORY PRACTICE COMPLIANCE STATEMENT Exygen Study Number Pooo{}76o, entitled "Analysis of Perfluorooctanoic Acid (PFDA) in Water, Soil, Sediment, Fish,, Clams, Vegetation, Small Marnmal Liver and Small Matnmal Serum Using LC/MS/MS for the 3M Decatur Monitoring Program," conducted for 3M Company, is being perfonn$d in compliance with EPA TSCA Good Laboratory Practice Standards 40 CPR 792 by Exygen Research, with one exception. The laboratory control and control spikes for the direct injection and alternate SPE method~ and the spiking procedures for the surrogate were not recorded promptly at the time of extraction. lmfl~ d'ohn Flaherty Principal Investigator Exygen Research Jaisimha Kesari P.E., DEE Study Director Weston Solutions, Inc. _1L Date 61_~Jo b _ _Da_te Exygen Research Page 2 of 110 000025 Interim Report #23 - Analysis of Sediment Samples Exygen Study No,: P'0000760 QUALITY ASSURANCE STATEMENT Exygen Research's QualitY'Assurance Unit reviewed Exygen Study Number P0000760, entitled, "Analysis of Perfluorooctanoic Acid (PFDA) 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 phasest were inspected for conduct according to Exygen Research's Standard Operating Procedures ; the Study Protocol; and all applicable Good Laboratory Practice Standards. All findings were reported to the Exygen Principal Investigator and Management and to the Study Director . Phase Date Inspected Date Reported to Date Reported to Principal Exygen Date Reported to Investigator Manag.ement Study Director 31 . Final Raw Data Review and Interim Analytical Report Review 512106 512106 512106 512106 a517-aWloa Date 'Note: All . in-lab inspections will be documented in the QA statement for the final analytical report at the conclusion of the study. This QA statement involves only the review of the interim report and associated raw data. Exygen Research Page 3 of 110 OooO2V lnterim Report #23 - Analysis of Sediment Samples Exygen StudyNo.: P{1000760 CERTIFICATION OF AUTHENTICITY This interim report, for Exygen Study Number P0000760, is a true and complete representation ofthe raw data. , Submitted by: Exygen Research 3058 Research Drive State College, PA 16801 (814) 272-1039 Principal Investigator, Exygen: ) Flaherty Date Vice President Exygen Research Exygen Research Facility Management : Date " Study Direcyor, Weston Solutions, Inc . Jaisimha'kesari P.E., DEE Weston Solutions, Inc. Sponsor,Representative, 3M Company: W310 ~ Dat J Date Exygen Research page 4 of 110 000027 _~., 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 Sum Using LC/MS/MS for the 3M Decatur Monitoring Program PROTOCOL NUMBER: P0000'760 EXYGEN STUDY NUMBER : P0000760 TYPE OF STUDY : Residue SAMPLE MATRIX : Sediment TEST SUBSTANCE : Perfluorooctanoic acid (PFOA) SPONSOR: 3M Company 3M Building 0236-01-B-10 St. Paul, MN 55144 STUDY DIRECTOR: Jaisimha Kesari P.E., DEE Weston Solutions, Inc. 1400 Weston Way West Chester, PA 19380 STUDY MONITOR : Michael A. Santoro 3M Company 3M Building 0236-01-B-10 St. Paul, MN 55144 PERFORMING LABORATORY: Exygen Research 3058 Research Drive State College, PA 16801 ANALYTICAL PHASE TIMETABLE: Study Initiation Date: 11105104 Interim Analytical Start Date: 04127106 Interim Analytical Termination Date: 04130106 Interim Report Completion Date: 05102106 Exygen Research f Page 5 of 110 000028 Interim Report #23 - Analysis of Sediment Samples Exygen Study No. : P004076Q ~ PROJECT PERSONNEL The Study Director for this project is Jaisinlha 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 CYespi Devon Ingrain Christa Gallant Title Vice President Scientist Technical Lead-LCIMS Technician Sample Custodian Associate Scientist Technician Technician Technician Technician Technician Exygen Research Page 6 of 1 1 0 000029 Interim Report #23 - Analysis of Sediment Samples Exygen Study No . : P0000760 TABLE OF CONTENTS RueTITLE PAGE ......... . . . . .... ............ . .... ...... .... . ............ .............. . . ..... . . . ............ . ..... . ............ . ... . ..I 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 ofStandards 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 . ....... ... . .... .. .... . ..... . ............. . .... . ...... .. .... . .. ....A7 7.0 EXPERIMENTAL DESIGN ... ..... ..... ..... .......... . .... ...... . ..... . .... ........ . . ..... . ..... .. .... . ...... ..19 8.0 RESULTS .... . ........ .... .......... . .. ......................... .... ...... ... . ...... .............. ...... ....... ..... ....... .19 9.0 CONCLUSIONS .... . .......... .. .. .......... .. ... ............ . .. ...... . .......... .. ...... . . ........... .......... .....20 10.0 RETENTION OF DATA AND SAMPLES ...... .. ....... ... ....... .. ..... ..... . ....... ... . . ...........20 Interim Report #23 - Analysis of Sediment Samples Exygen Study No .: P0000760 Table 1. LIST OF TABLES Page Summary of PFOA iri Sediment Sanagles . . ..., ....... . .......... . ........ .... . ..... ....... ..., .22 Table II. 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 ofPFCA in Sediment Samples . .... .. ...... . ..... ... ...... . .. ..25 Table V. Matrix Spike Recovery ofPF,OA in Direct Injection Sediment Samples . .... ..26 Table VI, Matrix Spike Recovery of PFOA.in SPE Extracted Sediment Samples .. .... . ..27 Table VII. Surrogate Spike, Recovery of '3'C~PFOA in Sediment Samples . ... ........... .. .. . ..28 Table VIII. ~Sunogate Spike Recovery of 13C PFOA in Direct Injection Sediment Samples ... ..... . ........ .... . .......... . .... .... ....... . ........... .........,........ ....... ... . ...... ..... ... . ....29 Table IX. Surrogate Spike Recovery of 13C PFQA in SPE Extracted Sediment Santples ........ . ........ .. .. ........... . ........ .. . ... . ..... ..... ....... ...... ....... .. .. ........ .... .........30 Table X. Total Percent Solids in Sediment Samples .. . . ......... . . ...... .... .. .... . ...... ........ . ....-31 Exygen Researub Page 8 of 110 Interim Report #23 -- Analysis of Sediment Samples Exygen Study No. : P0000760 Figure 1 . Figure 2. Figure 3. Figure 4. Figure 5. Figure 6. Figure 7. Figure 8. LIST OF FIGURES Page Typical Calibration Curve for PFOA in Methanol .. ...... . .... . . . ... ... ..... . . ... . ....... .33 Extracted Standards,of PFOA in Methanol, 25 ng/L and 50 nglL, Respectively..... . .... .,......... ........... .... . .......... . .... .. ...... ........ .... . ...... .... ........ . . ..... ...34 PFDA in Reagent Blank, 0.4 nglmL Fortified Reagent Spk A, and 4.0 ng1mL Fortified Reagent Spk B, Respectively...... . ...... .... .. . ...... . ... . . ...35 Chromatogram Representing a Sediment Sample Analyzed for PFOA (Exygen ID: Ct}172892, Data Set: 042806D) ............ . .... ........ ...... ...... . ...... .... .36 Typical Calibration Curve for 13C PFOA in Methanol..... ...... ...... .. .... . ...... .... .37 Extracted Standards of 13C PFC7A in Methanol, 25 ng/L and 50 nglL, Respectively .. . . ........... .. . . ........ . . . .. . . ........ . . . ........ .. .. .._. ...... . . . ......----.--... . . . .......... .38 13C PFDA in Reagent Blank, 0.4 nglmL Fortified Reagent Spk A, and 4.0 ng/mL Fortified Reagent Spk B, Respectively... . . ...... ........ .... . .......... .39 Chromatograrn Representing a Sediment Sample Analyzed far t3C PFOA (Exygen m- C0172892, Data Set: 442806D) . ..... . .... . . .. .. . . . ... .. ..... .4d Exygen Research Page 9 of 110 000032 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 LIST OF APPENDICES Page Appendix A StudyProtocol P0000760 (Exygen Study No. P0000760) with Analytical Method V0001782 : "Method ofAnalysis for the Determination of Perfluorooctanoic Acid (PFOA) in Sediment by LCIMSl1VIS", and Protocol Amendments ......... . ....... .. .. ... ..... . ............ . ..41 Exygen Research Page 10 of 110 O.()f0033 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 nglg . The standard sediment method (VOfl017S2) was originally used to extract all of the samples. Use of this method yielded poor spiking recoveries for PFOA. Because of 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 "C PFOA . Analytical results for the analysis of PFOA in sediment samples extracted according to the method 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 of PFOA in sediment samples by alternative SPE extraction, based on dry weight, are summarized in Table 111 . The overall average percent recovery t standard deviation for PFOA in sediment samples based on wet weight was 73 1610. Fortification recoveries for the analysis of PFOA in sediment samples are summarized in Table IV. Fortification recoveries for the analysis ofPFOA in direct injection sediment samples 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 '3C PFOA in sediment samples based on wet weight was 97 t 27%. Fortification recoveries for 13C PFOA in sediment samples based on wet weight are summarized in Table VII . Fortification recoveries for "C PFOA in direct injection sediment saniples 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 of this study was to determine levels of perfluorooctanoic acid (PFOA) in sediment according to Protocol P0000760 (Appendix A) . Exygen Research Page 11 of 110 000034 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 3 .0 INTRODUCTION This report details the results of the analysis for the determination of PFOA in sediment using the analytical methods entitled, "VOO01782: Method of Analysis for the Determination ofPerfIuorooctanoic Acid (PFOA) in Sediment by LC/MS/MS ." The study was initiated on November 05, 2004, when the study director signed protocol number P000fl760. 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 perfluomoctanoic 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 13C PFOA was atored frozen . COMMund PFOA 13C PFOA Exygen InventoryNo. SP0005444 SP0004184 Lot # ~ Purity (no 203 99 3507-195 97 Expiration Date 03I311U7 03129109 The molecular structure ofPFOA is given below : PFOA Chemical Name: Perfluorooctanoi .c acid Molecular Weight : 414 Transitions Monitored: 413 -~ 369 (for quantification) and 413 -4 219 (for confirmation) Structure : Exygen Research Page 12 of 110 oooQ3s Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 QH 13C PFOA Chemical Name: 1,2-13C perfluorooctanoic acid Molecular Weight: 416 Transition Monitored: 415 -)- 370 Structure : O F F F F P F 1 1}CII "" '3C~ OH F 6.0 DESCRIPTION OF ANALYTICAL METHOD The analytical method "VOOOl782 : 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. AS gam portion of sediment was weighed into a fifty milliliter centrifuge tube for the extraction. After fortification of appropriate samples, 35 mL of 19/o acetic acid in water was added to the samples. The samples were vortexed and allowed to shake on a wrist action shaker for-60 minutes. The samples were centrifuged for -20 minutes at -3000 rpm. The supernatant was then loaded onto a CI$ SPE cartridge conditioned with lq rnL 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 -2fl minutes at -3000 rpm. The supernatant was then loaded onto the same Clg 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 + ;~ra:{~: ~,~ Page 13 of 110 0ooo,3s Interim Report #23 -- Analysis of Sediment Samples Exygen Study No. : P0000760 hundred milliliters of water was added to the bottles. The samples were mixed by shaldng and loaded onto another C, 8 SPE cartridge conditioned with 10 rnL of methanol and 20 mL of water. The eiuate was discarded. Approximately five milliliters of methanol was added to the cartridge. Five milliliters of eluate was collected into a graduated 15 mL polypropylene centrifuge tube. Each sample was analyzed by LC/MS/MS electrospray. 6.2 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 of 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 of 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 -10 minutes at -3000 rpm. The supernatant was then loaded onto a C, s SPE cartridge conditioned with 10 mL of methanol and 5 mL of water. The eluate was discarded. Approximately five milliliters of methanol was added to the cartridge. Five milliliters of eluate was collected into a graduated 15 mL polypropylene centrifuge tube. Each sample was analyzed by LCIMSIMS electrospray . 6.4 Percent Solids Procedure For Sediment Percent solids were determined using the procedure indicated in Exygen method V000(}427. 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 ofthe pan. The percent solid for each sample was then calculated. See Table X for percent solids results. 6.5 Preparation of Standards and Fortification Solutions A mixed stock standard solution of PFOA and 13C PFOA was prepared at a concentration of 1000 pg1mL by dissolving 100 mg of each of the standards (corrected for purity and Exygen Research Page 14 of 110 00003'7 Tnterim Report #23 - Analysis of Sediment Samples Exygen Study No . : P00007fi0 salt content) in 100 mL of methanol . From this solution, the following fortification standards were prepared : Cone. of Stock or Fort. (S>,o~ lutiLo}n' 1000 100 10 1 .0 0.1 ~ of PFOA and Fort Volume (rnL) 10 10 10 10 10 13C PFOA Final Volume (mL) 100 100 100 100 100 Final Cone. of Fortification Std. 41glmL) 100 10 1 .0 0.1 0.01 A stock solution of only 13C PFOA was also prepared at a concentration of 100 pglmL by dissolving 10 rng the standard (corrected for purity and salt content) in 100 mL of methanol. From this solution, the following fortification standards were prepared : Cone. of . Stock or Fort. Solution (pg/r_.ml;, 100 10 1 .0 t of "C 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 t3C PFOA was prepared in methanol, as specified in Exygen method V0001782. The following concentrations were prepared Cone . ofFort or Calibration Solution Fort Volume (ML) 100 1 .0 100 0.5 100 0.2 10 1 .0 5.0 1 .0 2 .0 1 .0 1 of PFflA and 13C PFOA Final Volume (ML) 10 10 10 10 10 10 Final Cone. of Calibration Std . (ng/ML) 10 5.0 2.0 1 .0 0.5 0 .2 Exygen Research '~4z4~ ~ 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 nglmL 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 t 2C) when not in use. Docurnentation 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 of PFOA was -11 .1 min. Peaks above the LOD were not detected in any of the reagent blank samplds corresponding to the analyte retention time. 6.7 Instrument Sensitivity The smallest .standard amount injected during the chromatographic ran had a concentration of0.2 nglmL ofPFOA. 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 11 00 HP Quat Pump HP Vacuum Degasser HP Autosarnpler HP Column Oven HPLC Column: Thermo Fluophase RP, 50 mm x 2.1 mm Column Temp.: -30 C Injection Vol.: 15 uL Mobile Phase (A): 2 mM Ammonium Acetate in water Mobile Phase (B): Methanol Exygen Research Time &mdn) %A %B 0.0 65 35 1.0 65 35 8.0 25 75 10.0 2.5 75 11 .0 65 35 18.0 65 35 Page 16 of 110 p00Q39 Interim Report #23 -- Analysis of Sediment Samples Exygen Study No. : P0000760 Tqtal run time: -18 min Flow Rate: 0.3 mLlmin Ions monitored : Analyt e Mode PFOA PFOA Confirm Ion 13C PFOA negative negative negative 6.9 Quantitation and Example Calculation Transition Monitored 413--+369 413-+219 415-+370 Approximate Retention Time -11 .1 min. -11 .1 min. -11 .1 min. PFOA concentrations are quantitated in nanogram of PFOA per milliliter of extract and then converted to nanogram per grain. The' PFOA concentrations were rluantitated in nanogram of PFOA per liter for Set #3 (Set 042806D). Fifteen microliters of sample or calibration standard were injected into the LC/MS/MS . The peak area was measured and the standard curve was generated (using llx 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 (V4001782), but this has no effect on the final calculations . Equation 1 calculated the amount of analyte found (in nglL, based on peak area) using the standard curve (linear regression parameters) generated by the Analyst 'software program . Eauation 1 : Analyte found (nglL) = (Peak area - interce~t) x DF Slope Where: DF = Dilution Factor, factor by which the final volume was diluted, ifnecessary. For samples fortified with known amounts of PFOA and "C PFOA prior to extraction, Equation 2 was used to convert the amount ofPFOA found in nglL to nglg (ppb). uation 2: PFOA found (ppb) = tPFOA found {n&U x SPE volume extractedl fsample aliquot volume x (sample weight ! direct inject sample volume)] F,xygen .Research Page 17 of 110 000040 Interim Report #23 -- Analysis of Sediment Samples Exygen Study No.: P000076Q Equation 3 was used to calculate the percent recovery. Equation 3 : Recovery (%) = . (analvte found (nglgiwet weight) - analYte in control Lag/-& wet weight)) amount added (nglg, wet weight) x 100% 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 of PFOA found in ppb based on dry weight . Eauation 4: PFOA found (ppb) dry weight = PFOA found (ppb) x [ 100% I total solids(%)3 An example of a calculation using an actual sample follows: Sediment sample Exygen II? C0172892 Spk D (Set : 042806D), fortified at 4 nglmL with PFOA where: peak area = 236588 intercept - 4630 slope = 420 dilution factor = 100 nglg PFOA added (fort level) = 4000 amt in corresponding sample (nglg) - 586 SPE volume extracted (L) = 0.04 sample aliquot volume (niL) = 5 sample weight (g) _ 1 direct inject sample volume (mL} = 10 total solids (%) - 66.54 From equation 1 : Analyte found (nglL) From equation 3: PFOA found (ppb) = f236588 - 46301 x 1{y0 420 = 55200 nglL = (55200 nA x ~ 0.04 L) 15 rnL x (1 g I 10 mL)] .: . = 442.0 ppb From equation 3 : % Recovery = (4420 ng.,lg - 586 ng~g)- x 140% 4000 nglg = 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% l 66.54%) = 6640 ppb NOTE: This value may be slightly different than that ofthe raw data due to rounding. 7.0 EXPERIMENTAL DESIGN 13C PFOA was used as a surrogate for all the samples. l3C 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 concenthation 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. $.0 RESULTS Analytical results for the analysis of 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 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 t 16% . Fortification recoveries for the analysis of PFOA in sediment samples are summarized in Table IV. Fortification recoveries for the analysis ofPFOA in direct injection sediment samples are summarized in Table V. Fortification recoveries for the analysis ofPFOA 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 13C PFOA in sediment samples based on wet weight are szunmarized .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 Page 19 of 110 000042 Interim Report #23 - Analysis of Sediment Samples Pxygen Study No. : POOa0760 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 ail 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 of Sediment Samples Exygen Study No. : P0000760 TABLES Exygen Research Page 21 of 110 Interim Report #23 - Analysis of Sediment Samples Exygen Study No. : P00007b0 Table 1. Summary of PFOA in Sediment Samples Eacy9en Ip- C0172892 C0172892 Rep C0172893 C0172893 Rep C0172894 C0172894 Rep Client _ Sample ID DAL-5t}$PPIlt-0-D000 DAL-SD-BPPD7-0-DOOD' DAL-SD-BPP02-0-0000 DAL-SD-BPP02-0-0000' DAL-M-SPIP03-0-M DAL-SD-BPP03-0-0000" Analyte Found (ppb, nglB) Dry Weight CB Acid PFOA _ _ PerFluorooctenoic Acid NR NR NR. NR NR NR Assessed Accuracy {+/- yo) NR NR NR NR NR NR -Laboratory Duplicate NR = Not reported due to quality control resultiailures. Sea Table II for re-analysis results. Exygen Research Page 22 of 110 0po045 Interim Report #23 - Analysis of Sediment Samples Exygen Study No. : P0000750 Table YI. Summary of PFOA in Direct Injection Sediment Samples Exygen (D C0172892 C0172892 Rep C0172893 C0172893 Rep C017289+ C0172894 Rep Client sample ID DAL-SD-BPP41-0-0000 DAL-SD-BPP01-0-0000' DAL-SD-t3PP02-0-0000 DAL-SD-BPP02-0-0000' DAI--SD-BPP03-0-0000 DAL-SD-BPP03-0-0000" Analyte Found (ppD, nglg) Dry Weight C8 Acid PFOA Perfiuorooctanoic Acld NR NR NR NR NR NR Assessed Accuracy (+l- %) 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 ,_~`,~~t~kYS . Page 23 of 110 oooo4s Interim Report #23 - Analysis of Sediment Samples Exygen Study No . : P0000760 Table tIt. Summary of PFOA in SPE Extracted Sediment Samples on !0 C0172882 C0172892 Rep C0172893 C0172893 Rep C0172894 00172894 Rap Uborawry Duplicate Client Sample ID Analyb Found tppb, ng1g) Dry Weight Ca Aaitf PFOA Psrfluorooetanolc Add DAL-SD-BPPfl1-0-0000 881 DAL-SD-BPP01-0-0000 " 949 DAL-SD-ePP02-0-OD00 927 DAL-SD-BPP02-0-0000 " 823 DALSD-BPP03-0~9000 2390 DAL-SD-BPP03-O-OODO " 2380 Assessed Aecuracy ( +!- X) 30 30 30 30 30 3D Exygen Research Page 24 of 110 000047 Interim Report #23 - Analysis of Sediment Samples Exygen Study No. : P0000760 Table IV. Matrix Spike Recovery of PF4A in Sediment Samples C8 Add PFOA DAL-SD-BPP01-0-0000 (C07728s2 8pkD, 400 ppb8pake) DALSD-BPP01-G-0000 (C01728s2 Spit E, 4000 ppb Spike) DAL "SD-BPP02-0-0000 (p017t$ti3 Spk G,400 ppb Spike) DAL-SD-BPP02-0-0000 (C0172aB $pk H, 4000 ppb Spike) DAL-SD-BPPfl3-0-0000 (C01728w $pk J, 400 ppb Spike) DAL-SD-BPP03-1}0000 (Cfl7728s4 Spit K, 4000 ppbSpike) NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR Average : NR Standard Deviation- MR 'Sample residue exceeds the spiking level significantly (Sn spiking level); therefore, an accurate recovery value cannot be calculated. NR = Not reported dueto qualitycontrol result failures. See Table V for re-analysis results. Note : Sine* this sun"rnary table shows rounded results, recovery values may vary slightly from the values in Vie raw data. Exygen Research Page 25 of 110 000048 Interim Repart #23 - Analysis of Sediment Samp1es Exygen Study No.: P0000760 Table V. Matrix Spike Recovery of PFOA in Direct Injection Sediment Samples Sample ~ Description . - - DAL-SD" BPPd1-0-0000 (p017zsiQ Spk D, e0o0 ppb Spike) DAL-SD-BPP07-0-0000 (CO1TZeYZ S pk t:, 4oooo ppb Spike) DAL-SD-BPPa2-o-0000 (C0177ts93 8pk G, 4000 pph Spike) DAL-SD-8PP02-0-0400 (C017x8Y3 SpkH.40000 ppASpike) DAL-SD-BPP03-0 "0000 IC0172ss4 SpkJ, 9000 ppbSpike) DAL-SD-BPP03-0-0000 (C0172a9e Spk K, 40000 ppb B Pike) Spiked (Ogf9)} 4000 40000 4000 40000 4000 40000 C8 Add PFQA Amt Found Amount I in Sample Recovered (n9/g? we' weight __ (n9i9) wet weight NR NR NR NR NR NR NR NR NR NR NR NR Recovery t96} NR NR NR NR NR N12 Average ., NR Slaridard Deviation : NR 'Sample residue sxoeedsthe spiking level significanty, (Sx spiking l; evel) iherefore, an aocurate recovery value cannot be calculated. NR = Not repodled due to quality cwtrol result failures . See Table VI forre-analysis resuRs . Note : Slnos thls summarytable shows rounded results, recovery values may vary slightly from the values In theraw data. Exygen Research Page 26 of 110 000049 Interim Report #23 - Analysis of Sediment Samples Exygen Study No . : P0000760 Table VI . Matrix Spike Recovery of PFOA in SPE Extracted Sediment Samples CB Arad PFOA DAL-SD-BPP01-0-0000 (C017299Z $pk D, 4000 nplp Spike} 58$ 4420 96 586 23800 83 DAL-SD-BPP02-0-0000 (MyZt9d dpk 6, 4000 n01p dpike} DAL-SD-SPP02-0-0000 t0+172993 3pk H. e0060 npWn spike) 504 3880 84 504 25100 61 DAL-SO-SPP03-0-0000 IC017ZOY49pk J, 400 nplq SpIk*) 1080 4140 ' 77 DAL-SD-BPPO&D-0000 (C0171s94 Spk tt, aDOBO nyQ 8ptky y 40000 1 1060 23200 55 Average- 73 Standard Deviation : 16 Not*: Since this summary table shows rounded results, recovery values mayvary slightly from the vshres in the raw data. Exygen Research Page 27 of 110 000050 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.. P0000760 Table V11. Surrogate Spike Recovery of 13C PFOA in Sediment Samples Exygen ID Sample Description C0172892 C0172892 Rep C0172892 Spk C C0172$92 Spk q C0172892 Spit E C0172893 00172893 Rep C0172893 Spk F C0172893 Spk G C0172893 Spk H Ci1172$94 C0172894 Rep C0172894 Spk 1 C0172894 Spk J C0172894 Spk K DAL-SD-BPP01-0-0000 DAL-SD-BPPO1-0-0000 DAL-SD-BPP01-0-0000 DAL-SD-BPP01-0-DOa0 DAL-SD-HPP01-0-0000 DAL-SD-BPP02-0-0000 DAL-SD-BPP02-0-0000 DAL-SD-BPP02-0-0000 DAL-SD-BPP02-0-0000 DAL-SD-BPP02-0-0000 DAL-SD-BPP03-0-0000 DAL-SD-BPP03-0-0000 DAL-SD-BPP03-0-0000 DAL-SD-BPP03-0-0000 DAL-SD-BPP03-0-0000 Amount Spiked {nglg} 4 4 4 400 4000 . 4 4 4 40,0" 4000 4 4 4 400 4000 "C-PFUA Amount Recovered Wet Weight (n919) _ NR NR NR NR NR IMR "NR " NR ' . NR NR NR NR NR NR NR Recovery (yo) NR NR NR NR NR NR NR N R NR NR NR NR NR NR NR Average : NR Standard Deviation: NR NR = Not reported due to quality control result faflures. See Table Vill for re-analysis results. Note : Since this Summary table shows rounded results, recovery values mayvary slightly from the values in the raw data . Exygen Research Page 28 of 110 000051 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 Exygen ID 00172892 C0172892 Rep C0f72892 Spk C C0172892 Spk D C0172892 Spk E C0172893 {`.0172893 Rep C0172893 Spk F C4172893 Spk G C0172893 Spk H C0172894 C0172894 Rep C0172894 Spk I C0172894 Spk J C0172894 Spk K Sample Descriptlon . ------- DAL-SD-BPP01-0-0000 DAL-SD-BPP01-0-0000 DAL-SD-BPP01-0-0000 DAL-SD-BPP01-0-0000 DAL-SD-BPP01-0-0000 DAL-SD-BPP02-0-0000 DAL-SD-BPP02-0-0000 DAL-SD-BPP02-0-0000 DAL-SD-BPP02-0-0000 DAL-SD-BPP02-0-0000 DAL-SD-BPP03-0-0000 DAL-SD-BPF03-0-0000 DAL-SD-SPP03-0-0000 DAL-SD-BPP03-0-0000 DAL-SD-BPPD3-0-0000 Amount Spiked (WO) 40 40 40 4000 40000 40 40 40 4000 40000 40 40 40 4000 40000 1sC-PFOA Amount Recovered Wet Weight -Ing1g) 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-analysis results . Note : Sinoe this summary table shows rounded results, recovery values may vary slightly from the values In the raw data . Recovery (.6) NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR Exygen Research Page 29 of 110 000052 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.; P00007b0 Table IX. Surrogate Spike Recovery of i3C PFOA in SPE Extracted Sediment Samples Exygen 10 OW 72892 C0172892 Rep C0172892 Spk P 00172892 Spk E C0172893 C0172893 Rep C0172853 Spc G C.0172893 Spk H C0172894 C0172894 Rep 00172894 Spk J C0172894 5pk K sample DascripUon DAL-SD-BPPDI-0-0D00 DAL-SD-BPP01~0000 DAL-SD-BPP01-0-0000 DAL-SD-BPP01-0-0000 DAL-SD-BPP02-0-0000 DAL-SD-BPP02-D-0000 DAL-8D~BPP02-D-0D00 DAL-SD-BPPD2-D-0000 DAL-SD-BPP03-0~0000 DAL-SD-BPP03-0-0000 DALSD-BPP03-0-0ODO DALSD-BPP03-0-0000 Amount Spiked (nglg) 40 40 40DD 40000 40 40 4000 40000 40 40 4000 40000 N: ote Sinca this summary table slwvrs rounded rewlts, rocoveRy YWU03 may very slightly from the values In 1ho raw data . ,sCrPPOA Amount Recovered Wet Weight ~~0~ 32.0 51 .0 3750 26800 47.8 51 .8 3500 25200 45.2 40,3 3060 22100 Avenge: Standard Deviation; Recovery (yb) 130 128 94 85 120 129 88 63 113 101 77 55 97 27 Exygen Research Page 30 of 110 4)00~<J+~ hxterim Report #23 - Analysis of Sediment Samples Exygen Study No . : P0000760 Table X. Total Percent Solids in Sediment Samples Exygen ID C0172892 C0172893 C0172$94 Sample Description DAL-SD-BPP01-0-0000 DAL-SD-BPP02-0-0000 DAL-SD-BPP03-0-0000 Total Sollds (96) 66 .54 54 .37 44,48 EXygen Rcsearch Page 31 of 110 000054 Interim Report #23 - Analysis of Sediment Samples Exygen Study No . : P0000760 FIGURES Exygen Research Page 32 of 110 000055 p. 33 Interim Report #23 -- Analysis of Sediment Samples Exygen Study No.; Pd00076U Figure l. Typical Calibration Curve for PFOA in Methanol oa2eoeDS.e .SPE .rdp(PFUIq)!'tJn*rrR.p149ricn("lrxwr~yMiny7y~42Dx+4.03~+U03P~o .DCp6) 42eff ~ 4AaS " 3.8e6 ~ 3Ae5 " 3.4s5 4 3.2e6 3.065 ~ 2"~ 2.e .a 2.4W- u 2.2~b m 2.Ue8 ~ a 1 .6~6 " 1 .4~6 1 .2~5 " Exygen Research 't i ) Page 33 of 110 000056 Interim Report #23 - Analysis of Sediment Samples Exygen Study No. : P00007b0 Figure 2. Extracted Standards of PFOA in Methanol, 25 nglL and 50 ng/L, Respectively , xcORs+r.1 -Pr-Os tSrarAe4* 4xtru3ia 8me .*awpk 2oraaoW ansnen,,wrr Ilres: 7lZ:i111coartr Mv~h L10&+Mqw PT. Tl.SOfu +1 .D1 w a t i 0 6 7 B p 1D 11 12 13 14 15 16 17 Time, min , ?LC04260-2 ~ PFOA (Standard) 413 .DI36DA amu " sample 3 q440 from 04280ODmiff Atom, 20159 eouni's Nllpht 2 .14f4.Ob2 o0et RT :11 .1 min 1105 N a u c Exygen Research Page 34 of 110 OoOO5'7 Interim Report #23 -Analysis of Sediment Samples Exygen Study No. : P0000760 Figure 3. PFQA in Reagent Blank, 0.4 nglmL Fortified Reagent Spk A, and 4.0 ngImL Fortified Reagent Spk B, Respectively , Reryewt Cartrol-PFQA tMrtrowir)4736I3ifL "asr -sawA ! oT4i!ls6o IK2"Oi.0w!!P drest wottowrd) a V W C 41 15 2 3 q 6 a 7 8 6 10 11 12 13 14 16 10 17 Time. min , Riapeni Sph A- PFOA (QC) 413At360A amu " sample 10 af 46 hom Oq'28060.wH1 Area : 30296 aourrIF Height 2 .370+003 opc AT : 11 .0 min m a Z0M , 11 .00 c 1000 1 , io .ao,, ~ I 2 .00 7,66 8 .60 ~---~,~~ 13.80 0 1 3 3 q 6 Q 7 0 0 10 11 12 13 14 15 16 Time, min Reagent BpkB-PFOA(QC)413Aq6D,Damu-sampte 11ot40irom04280G0 .wiH Area : 260300 oourrtc Height 2 .OCe+004 cps AT: 11 .1 min Tlmc min Exygen Research , . Page 35 of 110 000058 Interim Report #23 -Analysis of Sediment Samples Exygen Study No. : P004076U Figure 4. Chromatogram Representing a Sediment Sample Analyzed for PF1JA (Exygen ID: C0172892, Data Set: 042806D) , G6172092 "PFOA 6"+rawr)fWAft"as,v-smrpli 79sf4ttrorr 04l9rsWxrlli Area. 33d2fcowts JWgit: Lf6r+0A1 yos RP ".0 aN+r 17n6 N o u . c Exygen Research , Page 36 of 110 OOO055 p. 37 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: Pil0U7b0 Figure 5, Typical Calibration Curve for 13C PFUA in Methanol 0428o6D s .a- SPE.rdD (13E PFOft " unnar Raprmlen C"I i,r,onlghtlep): y=40c x+v .oolv (r y O.ooo)) 9.0a6 3.445 3.2@6 3.046 2_Be61 2 .0@6 0 2 .4a8 d 2 .20a- 2A*b ~ 1 .905 1 .D86 1.4`6 1 .296 1 .O45 8.064 0.0A4 " 4.0642.Oa4- 0 .0 ~ ConoonuaCon.nplL Exygen Research Page 37 of 110 OOQO60 Interim Report #23 - Analysis of Sediment Samples Exygen Study No. : P0D00760 Figure 6. Extracted Standards of 13C PFOA in Methanol, 25 ng/L and 50 ng/L, Respectively , XGN200f+! -73C PFQA ~a~d++~4'!~,/A7a0aw~r .soaP1r2 of416 tro+r 01200601WfJF prQa : tft0ye,ywt: NbIQAU 7,lAv+003 cps R7:11.0 srJx 11.02 1001) m 800 n k BOp tc1 w c 4DD 1 2 3 4 6 e 7 8 0 10 12 13 14 i5 10 17 Time, min ! Xt042606-2-13CPFOA(StanQard)415A1C47D .Oamu " sampIO3ot401rom042808D.ni11 " Arm 26B9B oouob HoipM :2.46r,*003 ops R7: 11 .1 mla 11 .00 tu CcaL c m Exygen Research Page 38 of 110 O00O61 p. 39 Interim Report #23 - Analysis of Sediment Samples Exygen Study No . : P0000760 Figure 7. 13iv PFOA in Reagent Blank, 0,4 nglmL Fortified Reagent Spk A, and 4.0 ng/mL Fortified Reagent Spk B, Respectively , Rergaa! Coatrni " t3CPPoA "*now) 4134rJM4s+nv -*awpk Yol4itrow W2BOiAwf11 #tA1 1110 terRd) m a c 0 D 10 Time, min Reapenl Spk Jl- 13C PFOR (OC) 416.Ot370 .0 amu " srmple 10 af 461rom 0428000 .witt ' Arem :2729p counts Heiyht2 .02e+003 cps RT : 11 .0 min var . ca 2000 - 11111 cm 1000c 0. 1 2 4 -'-'1 7 B 9 10 11 12 13 14 1iG 10 17 r Time, min RupeM Bpk B - 13C P F OR (OC) 416.Ot37D.0 amu - "mple ! 1 of 48 from 0420p6 Di+ri$ Arna :270Q01 counts Height 2Ms+004 cps RT : 111 .1 min m Q 2Ae4- ' c 1 .0e4 " 11 .07 0,0 1 2 3 A 5 0 7 8 0 10 11 12 13 14 1d 18 17 Time. min Exygen Research Page 39 of 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) , Co9lxK92 " y3GPFt7A (LkrpoWlr)4r3AMe ..rer .sserph 27ofd011ver 0421rR.0sW1I Arm:3W30 eorrrtr hY1gft 2910+Oq4cpe R7," ?f,irrin 2 .064 . 47 .07 2.0a4 2.4e4 , 22.B4 2.0r4 1 .9a+1 V1 V 1 .Oa4 N c 1 .444 ar 1 .2e4 1 .0&4 8000 .0 000010 4000A 2000.0 0.0 1 2 3 4 6 0 7 B 9 10 11 12 13 14 16 10 17 Time . min Exygen Research Page 40 of 110 040()63 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 APPENDIX A Study Protocol Pd000760 (Exygen Study No. P00007b0) with Analytical Method and Protocol Amendments Exygen Research Page 41 of 110 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P000a760 Exygm Pmmooi Number. >M0760 STUDY PROTOCOL Study Title: Analysis of Pertluorooctanoic Acid (PPOA) in Water, Soil, Sediment, Ftsh, Clams, Vegetation, Small Mammal Liver and Small Mammal Serum Using LC/MS/MS for the 3M Decatur Monitoring Program Exygen Protocol Number: P0000760 ggrfomiing Laboratorv: Exygea Research 3058 Research Drive State College, PA 16801 Phone; (814) 272-1039 Sponsor Rermsentatiye : Michael A. Santoro Director of Regulatory Affairs 3M Building 0236-01-B-10 St . Paul, MN 55144 P: hone (651) 733-6374 rag. r ofas Exygen Research Page 42 of 110 000065 Interim Report #E23 -- Analysis of Sediment Samples Exygen Study No.: P00007b0 Exygen Protocd Number POOD0760 DISTRMUTIUN: 1) Jsiaimha Kaari, Study Dirxtor, Westott Solutions 2) John M. Plaherty, Principal Investigator, Exygen Reaoarch 3) Michael A. Ssntoro, Sponvor Represemtative, 3M Company 4} Fxygen Reaearch Quality Ascuuanoe Unit Exygen Research page ? of 6J Page 43 of 110 OOOOGG Interim Report #23 -- Analysis of Sediment Samples Exygen Study No.: PUUOfl760 Bxysaa Protocol Nwnber : PD00o75v 011- PROTOCOL APPROVAL Study Title: Analysis of Perfluorooctanoic Acid {PFOA} in Water, Soil, Sediment, Fish. Clams, Vegetation, Small Marrrnal Livers and Smafl Marnmal Scrum Using LCIMSIMS for the 3M Decatur Monftoring Prograrn Exygen Protocoi Numbar: POOO0760 APPROVALS Jaisim$`a Kcsari, Study Director Weston Solutions ., x ////b patc M. Flaherty, Principal Investigator Exygen Research ichud A. zini, ExYgen Rose64 sident, Facility Management 1~~~~ Date u3 -aT -740t/ IJat9 ate 0-1 Pqw3 of 6J Exygen Research Page 44 of 114 oo~a()s7 Interim Report #23 - Analysis of Sediment Samples p. 45 Exygen Study No._ P0000760 Exygen Protocol N=W : P00007b0 TABLE OF CONTENTS TITLE PAGE ._.._ ..... . . .. ... ._ .... .. . .. ... ...... .. ._ .,. .W..... .. . ... . .... .:. .,...., .. ... . .. .... .. ...., .. ..... . . ..... . .. .... . . .,.. ... . ..... . ., . .. . .. ....1 DI87'RIHU710N .. ..... . . .. ._ . .. ... ... . .. ... ...... .. . .. . .. ..., ... . . . .. . .. . . .._. .. . .,...., ., ......_.. .. ., ..... . .._. . ., . .., . .. . ...,. . . .. ... . ..,.. . . . ....2 PROTOCOL APPROVAL, .,.. ... . .. ... .. ... .._ ., .... .. . ..... . . ._. .. :. ...__ .._._ . . . . ...., . . ._ .. . . .. .,: ._ .:. .., . .. . . .. . .. . .. . . .. ... . . . ... . . . ... .3 TABLE OP CONTENTS . . . . ..... . . . ....___-___.. . . .. ... .._ . .. . . . . .._ . .._ . . . . .... . . . .. . .. . . ...... ... ... . .. . ., . . . . . . . . . . ._ . . ._.. .. . ... .4 INrRODUC'YiON .. . .. . ...... .. . . .... . .. . . . ._ ..__ .._._ . ...... .. . .. . .. . . ... .__ ._. . . . . . ._. ._ . . .... ._ . ._ . .. . ... . . . . . . ... ._. ._ . . . .. ._. .5 TEST MATERIA[., ... . .. .... .. . ... . ...., .. . .. . .. ...... ... .. . ........ . .. ... ..., . ...... . .. .. . .._. . .. . . .. .,.. .. ., .,.. . .. . ..,.. . . .. ... . . . .,.. . .. ..... . . .. ._ . .5 SURROGATE FIELD SPIKE COMPOUND ... . .. ...... . .. .. ._ .. . ...... ., ... . .. ... ... ........ .. .. . ._ ... . ....: . . . . ... . .. . ... . . . ... .. . . .. ... . .5 OBJECTIVE . ..._ . . .. ._ . . . .._.__ ..._ .. . . .. ._ . . .... .. . .. ... . . . ._ .. . . .... . .. . ...... .. .. . . . . ... ..-..._ . .... .. . ..,. . . . . ._. .. . . .._ . . .. . .-. . .. . .. . . .. . ..- .6 TESTING FACILI7Y . . ._ .. . .. ... . .._ . . .. ... . ... .. . .. .., ., . .. .,. . ...., . .... . .. .... ._. . . . .... .. .... .. . .._. . . .. ... . .. .. ... ... ... . ...--- . . . . .. . . .., .. . .6 STUDY DIRBChOR . . . . . . . . . .. ...: .. .. . .. . ... .. ... .. . ...... . . . . . . . . . .... . ....- . .. .. . ..._. . .. . . .. .... .. . ..-.. .------" --- ... . .._ ._. . . . . .. . .------------.6 SPONSOR REPRESENTATIVE. . .. ..... ... . . .... .. .. . . ..... . .. .. . .. ... .... . . . . ... ... . .. .. . . . . . .. .... . .. ... . ... ... . . . . .. . . . . . . .... .. . -7 PRINCIPAL INVESTI4ATOR ... . . ._ . .. . .. . ... .. .. ... ... . ... .. .. ., ..,.. .... . . .. ... .... . . ., . .. . .. .. .. .. .,., . .. .. . . .., .. . ., . . .. .,. . . .. . .. . . . ... . .. . ., .7 PROPOSED EXPERIMENTAL START AND TERMINATION DATES . .... .. . ... . . . .. . . . . . .. . . . . . .. . .. . . . . . . . . . . ... . . . . . . . 7 IDENTIFICATION AND )USTIPICAT10N OF THE TEST SYSTEM .. ... .... .. . .. .. . .. . .. . . .. . . . . . .. . . . . . . . .. . . . . .. . . . . . . . 7 SAMPLE PROCLIREMENr, RECEIPT AND RBTS?JTION . .. .. . .. . .. . .. .... .. . ..:... .. . ., ., .. . .. . ... . . . . ... . . . . . . .. . ... ... . .. . . . . .8 SAMPLE IDENTIFICATION . .. . ..-. . ..-. . .,. ... . . . . .. . . . . . . ... . .. .. . .. ... . .. . . . ., .,., .. . ... .. . . . . .. . .. . .. .. .. . .. .. .. . . ... . . . . . .. . . . . .. . . . . . . . .. .- . 8 ANALYTICAL PROCEDURE BCTA9v1ARY . .. .. .. . . . . .,. . . .... . . . .. . . ... . . . . .. . . ..,.. ... ... ..., .. . .. ..., . ..... .. . ... . .. . . .. ., ., ... .. . ...... ., 9 VERIFICATION OF ANALYTICAL PROCEDURE. .. .... . . . .. . .. .. . . . ... .., ., .. . .. .. . .. ., .. . .. ... ., . ... . .. .,.. . .. . .. . .. . . ., . ., . ..-. . . . . g MSI'HOD FOR CONTROL OF BIAS .... . .. .. .. . . . ..... ., . ..... . . .. .. .. . .. . ... ... . . . . .. . .. . .. . ... . .... . .. ... . . .. . .. . . . . . .. . .. . . .. .. . . .. .. . . . .11 STATISTICAL METHODS ... ... . . . . . . .. ...... ._ .. ..... . ...._ .. . .. ... . .. .- . .. . .. ...-. .-.. .. .--- . .. . .. . ..... .- .-------------- . .__ . .. . . . . . .-------1 i GL.P STATEJNENT . .. . . .- ... .. . ... .. .- . . ... . . ... .. . ._ .. ... .. . . . .... . . . .. .. . ._ .. . .. . .. . .. . . . ... .. . . . . .. . . .. ... . .. .. . . .. ... . . . . .... .. .... ., . ... .. . . .-. .- . .- .1 l RE$ORT . . . .. . .. . . . . . . . . .. . ... ..... . ..... . .. .. . . . . .._. . . .... ... .. . ..... . .. . .. .. . . ... . . .. .. . ... I I SAFETY ANDHEALTH ... . .._. . .._.. . . .._. .. .... .. . .. . ..... ... . .. .. ..... . . .. ... ., . . .. .. . . .. .. . . ..... . ....., ., . .. . .:, ... . .. .... . . . ..... . . .. ... . . . . . 12 AMENDMENTS TO PROTOCOL .,_. ..... . .. . .. . ..____ .. ._.. . ... ... . .r._ .... .. . . ...., . .: ... . .. : :.: ...... ... .... .. .,.... . . .. .. ., ., 13 DATA RECORD KP.EPINU . ... .... . . .. . ...... .. . .,. ... .. . ...._ .. ... .. ., . . .. .., ., .._ .. .. .. .. . .:. .. . . .._. . .. . ....._ . . .. . .... . . . .. . . . . . .._ . . .. . .13 QUALITY ASSURANCE .... . .. . ..... . .. ... ...,.... ... . ... .. . ..... ... ... .. .... ... .. . . .._.. .. . ._ .. .. ..__.. .._.. ... . .. . .. . . .. . .. . . . . . .._ . ... . .14 RETENTION OF DATA AND ARCHIVING . ...... . .. ... ._. ._____ .._. . . ._.. ._ . .. . . .._. . .._.._. . .. . .. . . .. ... . .. ...._ . ... ..14 APPENDIX I . ANALYTICAL METHODS ..... ... . . . . ... .. .. . ...._ .. __..... ... ... . . . . . .. . . . .. ..... . ... . . . .. . .. . . .. ... ... . .. . . . .. . ] 5 101Pqge 4 ofES Exygen Research Page 45 of 110 0000w Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Exyyen Protxol Natobe:: P0000760 INTRODUCTION The purpose of this study is to perform analysis for perfluorooctanoic acid (PFOA) in water, soil, sediment, fish, clams, vegetation, small mammal li vers and small mammal serum using LCIMSlMS for the 3M Decatur Monitoring Program. The study will be audited for compliance with EPA TSCA Good 1.aboratary Practice Standards 40 CPR 792 by the Quality Asaurance Unit of Fxygen Research. TEST MATERIAL The teat material is perfluotuoctanoic acid (PFOA) and was purchased from Sigma-Aldrich. PFOA Chemical Nam: Perfluorooetande add Molecular Weight : 414 Lot Number. 2311dHB r Purity; 97.64% Transitions Monitored : 413 -+ 369 (far quantification) and 413 -+ 219 (for confirmation) Structure: SURROGATE FIELD SPIKE COMPOUND The surrogate field tipiking compouud is `3C labeled perfluorooctanoic acid (t ;C PFOA) aad was purchased by 3Nf f:ora Perldn-Eltner Life and Analytical Sciences. '3C PFOA Chcmical Namo : 1,2-13C perfluorooctanoic ticid Molecular Weight: 416 Lot Number: 3507-195 Pusity: 97k Transition Monitored : 415 --~ 370 Page 3 of as Exygen Research Page 46 of 110 0000469 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 &xysen vnowro:Nwnb: POaoo760 OBdECTiVE The purpose of this study is to perform aualysis for perfluorooctanoic acid (PFOA) in water, soil, sediment, fish, elams, vegetation, small mammrl livers and small mammal sentrn for the 3M Decatur Monitoring Program using thc current versions of the following Exygen unalytioai methods : V0001780: Va001781 : V0001782: V0001783 : V0001784; V0001785 : V0001786 ; "Mcthod of Analysis for the Determination of Perl3uoroocianoic Acid (PFOA) in Water by LCtMS1MS" "Mdhod of Analysis for the Determination of Peffluomoctanoic Acid (PF(}A) in Soil by LCJMSlMS" '9Vletltod of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Sediment by LC/MS/MS' "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Fish and Clams by LGMSIM S" "Method of Analysis for the Dotermination of Perfluoraoctanoic Acid (PPOA) in Vegetation by LCIIvISIMS" "Method of Analysis for the Determination of Pcrfluorooctanoic Acid (PPOA) in Small Mammal Lver by LGMSIMS" "Method of Analysis for the Determination of Porfluorooctanoic Acid (PFOA) in Small Mammal Serum by LC/MS/MS" 1-11 TESTING FACILITY Exygat Rasearch 3058 Research Drive State Collegq PA 16801 Phone: (814) 272-1039 STUDY DIRECTOR Jsisitnha ICesati P.E., DEE Weston Solutions, Inc . 1400 Westoa Way Wag Chastar, PA 19380 Phone: (610) 701-3761 Fax: (610) 7D1-74E11 j .kesariowesionsolations.eom 0-1 Paga 6 q65 Exygen Research :v~}ltk:~l~ I Page 47 of 110 Interim Report #23 -Analysis of Sediment Samples Exygen Study Na. : P0000760 BxygpnProtocolNumber. POO00764 SPONSOR itEPRESENTATM Michscl A. 5antoro 3M Company Director ofRegulatory Affairs 3M Building 023"1-3-10 St. Paul, MN 55 144 Phone: (651) 733-6374 PRINCIPAL INVESTIGATOR John M . Flahetty Exygen Research 3058 Rosomb Drive State College, PA 16801 Phone : (8 14) 272-1039 john .flahertycsygan .corn PROPOSED EXPERD4EAITAI. START AND TERhMATION ,a- DATES It is proposed that the anafytical portion of this study be conductod from October Ol, 2004 to Dcocmbcr 31, 2005 . 'the actual experimental start and termination dates will be ineluded in the ftnal report. IDENTIFICATION AND JUSTIFICATION OF THE TEST SYSTEM The following ere thc test system for this study: i Water (groundwater and surface water) . soil . Sediment . Fish " clem " Vegetation Small Mammal Liver " Small Mammal Serum The samples will be collected by We9ton Solutions. The control samples wil l 10-1 be purchased and prepared by the testing facility. Purchase and processing Pop r qfa5 Exygen Research Page 48 of 110 0000'71 Interim Report #23 - Analysis of Sediment Samples Exygen Study NQ. : P0004760 F.iygen Protocol Number. POOW760 r- details for the catttrol samplcs will be included in the final report associated with this study. The test systems were chosen to access the environmental impact of PFOA in the Decatur, Alabama area . SAMPLE PRUCURIMNT, 29CWT .AND RETRNTION Water, soil. sediment, fish, clam, vegetation, small mammal liver and small mamnul serum samples will be received at Exygen directly ftam Weston Solutions. The details of sample procurement for this study are outlined in the 3M work plan entitled "Pima 2 Work Plan for Sampling Environmental Medis." The number and types of samples collected will vary depending availability in the fiold. 77re total number of samples received and analyzed for each matrix will be documented in the final report associated with this atady. Water, soil, and sediment samples will be used as received without further prooeasing at Exygen. These samples wiII be stored refrigerated at 2C-8C . Fish, ohm vegetation and small mammal liver samples will be processed according to the appropriate analytical method (see Appendix 1). These samples will be stvnd frozen at 5 -1Q'C. Small mammal whota blood samples will be centrifuged in the field at the time of collection and the serum fraction will be used for the study. Small mammal serum will be stored frozen at 5 -10'C. The receipt and processing of the samples will be documentod in the final report and raw data associated with the study. SAMRLE IDENTtFICATION Prior to analysis, cub sample will be assigned a laboratory sample reference umber. The reference number will be unique and will distinguish each laboratory sample due is processed throughout the analytical procedure. Chromatographic data will be identified by tha laboratory sarnplo raference number. Sample storage cottditions and locations will be documentod throughout the aa,ay. 10-- Page 8 o/65 Exygen Research Page 49 of 110 000072 IntetYm Report #23 - Analysis of Sediment Samples Exygen Study No . : P00007b0 ExyrYen Protocol Namher. PODD076o 00-1 ANALYTICAL ]PROCEDURE SUMAARY References: V0001780: V00017B 1 : V0001782: V0001783 : V0001794: V0001785: V000178G: "Method of Analysis far the Determination Of P0ff1uOT0OCl3noiC Aaid (PFOA) in Water by LClMSThiS" "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFaA) in Soil by LC/MS/MS" "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Sedirnent by LCIMSIMS" "Method of Analysis for the Determination of Perfluorooetanoic Acid (PFOA) in Fish and Clams by LC/MS/MS"Method of Analysis. for the Determination of Perlluorooctanoic Acid (PFOA) in Vegetation by LCAISIMS" "Method of Analysis for the Determination of Perfluorooetanoic Acid (PFOA) in Small Mammal Liver by LCJMSIM S" "Method of Analysis for the Uotermination of PcrEluorooctanoic Anid (PFOA) in Snma11 Mammal S== by LC/MSA4S" The above methods use analytical conditions capable of separating the isotues of PFDA, Thc final report will include the isomers atttnmed into total PFDA found. VERIFICATION OF ANALYTICAL PROCEDURE A laboratory control sample will be used for the preparation of fortified control samples. The test substance will be made into solutions as per the metbodc and added to the matricea via a micropipcttc . Forwater sampling, Exygen'will supply one bottle .per sample collected. The bottles will be- 500 mL prexleaned . ScilSpec Premier wide mouth HDPE bottles. . Thesebottles have been t+autinely used far PFOA sample collection at the teeting facility uxi have been shown to be frae of PFOA . All containers used for water sample collection will be shipped to the sample location contaihing 190 ng of t3C PFUA (equivalent to 500 nglL in the final sample). Sarnplea wigba addedto each contsinar so a volumetric fill lina at 200 mL. A field duplicala, a low field spike and a high feld spike of each sample will be collected. The low and high field spikes will contain PFOA as well as perfluornbutttnestalfonate (PFt13S), pertluotvhexanesutfonate MS), and perfluorooctanesulfonate (PF06). These compowuis are included in the solutions used to spike !he samples. The results for PFBS, PFHS and FFOS I" will not be reported in this study. , Exygen will supply one field blank (control water) and two field blank spikes (control water fortified with PFOA at a low Page 9 oJ 65 Exygen Research Page 50 of 110 000073 Interim Report #23 - Analysis of Sediment Samples p. 51 Exygen Study No.: P0000760 Psxygam Protocnl Number: P0000760 . ., and high level) for every tweaty samples collected . At the testing facility, each water sample (oxehiding fled duplicatea and field spikes) will be extracted in dupScate and will also be fortified at a low and high concentration with PPOA and processed thmugh the described procedure to determine method accuracy and to check for bias. For soil, sediment, clams, and vegotstion. Exygen will supply one 500 rnL precleaned ScilSpec Premier wide mouth FDPE bottle per sample collected or a zip-seal bag. AN containetg6bags used for ample collection will be shipped to the sample location . .Samp3es will -be added to each container or bag in the S:ol'd At tho testing fiurdIity, each sample will be extracted in duplicate and will also be fortified at a known concentration with PFOA at both a low and high level and processed through the described procedure to determine tn4Mhod accuracy and to check for bias . 13C-PFOA will also be added to each sample in the laboratory prior to extraction at the level specified below . For small mammal liver, Hxygen will supply a 50 mL polypropylene centriiyge tube. Tor small tnammal saruun, Exygon will supply a collection kit for each sample containing serum separator tubes (red top), vacutainers. needle holders and needles, transfer pipettes, end polypropylene tubes. At the testing facility, each liver and serum sample will be extracted in duplicate and will also be fortified at a known concentration with PFOA at both a low and high level and proceued through the described procedure to detcrmine method accuracy mid to check for bin. 13C-PFpA will al3o be added to each sample in the laboratory prior to extraction at the level specified below. Low and hi levela for each matrix Matlix Low PFOA Sp iking Level Water 500 n Soil 4 n Sediment 4 Fish 10n l0n V etaiion 10 n Small Mammal Li+ror 10 n Small Nisannipl are defined below : High PFOA S ikln Level S 5000 n 40 aWg 40 100 n 100n 100 n 100 n _ C-PFOA ikiu Level 500 n L 40 n _ 40 n 1Wn 100n 100 n 100 n Reaoveries are anticipstad to-be between 70V. and 1309C of the Fortified levels- however, The exact proGlaiota and accuracy will be dcteratincd by the analysis of the -quality cantrol sattrflles described abova A statement of accuracy will be included in the final report . Prior to aanipla Arialysis, a lahoratory control apike study will be conducted in water to justify the we of "C PTrqA results to establish precision and acetuacy of PFOA samples. ll!te following samples are to be prepared : Page lo o16s Exygen Research Page 51 of 110 000074 Interim Report #23 - Analysis of Sediment Samples Exygen StudyNo.: P0000750 Sxygen Promcol Number P0000760 e 'il+vo control waW blanks Two control water uunples fortified at 250 ng/L with "C PFOA a Two control water samples fordficd at 250 ng/L with '}C PFOA and at 250nglL with "OA. tr Two coattol water samples fortified at 5000 ng/L with "C PFOA and as 500p ng/L with PFOA The above uttrtplew are to be axtraebed and analyzed according to method YOD01780 entitled "Mcth4 of Analysis for the Determination of PerlYuotooctanoic Aeid (PFQA) in Water by LOMSIMS.' METHOD FOR CONTROL OF SiA8 Control of bias will be addressed by taking rtlmseqtative sab-samples from a homogeneous mixture of each matrix from untrested control aarnples, and by mlalyzing at least twolevels of fortifications . STATISTICAL 11IETH0IDS r~ Statistics will be limited to those specified in the subject methods and to the calculation of average recoveries, as applicable. GLP STATEIVIENT All aspects of this study shall be performed and reported in compliance with EPA TSCA Good Laboratory Practice Standards 40 CPR 792. The final report or data paekage (supplied to the Sponsor) s1taA contain a statement that the study was conducted in compliance with eurrettt and applicable GLP standards and will outlino any deviations in the study from those standards . This etsternent will be signed by the Study Director and Sponsor Reptpsentative . REPORT A final report will be prepared by the principal investigatar or their designee at the conclusion of the study. The report, will include, but will not be limited to, the following: Page I1 of 55 Exygen Research Page 52 of 110 000,0"-4,5 Interim Report #23 - Analysis of Sediment Samples p. 53 Exygen Study No. : P00aa'lso ExyW 1+mtocol Number : ffl00076D n The name and addtrsa of the Study Director, Sponsor Representative, and of tha testing facility. . A statement of GLP compliance (any related documentation, such as chain-of-eustody records, mast be in the study records}. e The signed and dated statement by the Irxygen Resoarch Quality Assurance Unit regarding dates of study inspections and dates findings were reported to the Study Diroetor and Management. 0 A description of the exact analytic-at conditions eatployod in the study. If the subject method was followed exactly, it is necessary to include only a copy of the analytical tnathod. Any modifications to this method will be incorporatod into the report . If the method is photo-reduced, the project number and page number must be included on each page, Description of the. itutttmtentatlon used and operating eonditions . . All results from all sets analyzed. Control and fortified samples will be identified and the data table will include sample number and fortification level. Representative chromatograms for each analyte in each matrix, including chromatograms ofa standard and a control sample, and a cfuomatogram at a fortification level. The location of the analyte peaks will be clearly identified in all ehromatograma . ~ All circumstances that may have affeccted the quality or integrity of the data will be documented in the report. Locations wharo raw data and the final report are to bo archived, a 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 of the report that is being altered and the reasons for the alterations. Tbe atneauLnent will be signed and dated by the Study Director andtheSponsor Representative. SAFETY AND HEALTH Laboratory personnel will 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 rCferencc subttancc(:). Page !1 of6J Exygen Research Page 53 of 110 Og'907s Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P000076{} Exygeat ProrocoY Numbec: P000o7W r` AmE1VDMENTSTOFRQII'OCOL Ali significant changes to the analytical protoeol outlined hero will be cxprassod in writing, aigaed and dated by the Study Diroctor and Sponsor Itapresentativc . Amendments usually will be issued prior to initiation of study plan change. However, when a change is required without sufficient time for the issue of a written atnatdment, that change may be effected verbally with supporting documeitation signed and dtttod by the Study Director and followed with a written amendment as oom as possible . In this case, the effective date of the written amendment will be the date of the documented chaaga. Copies of6c signed sQaendmemts will be appended to all distributed study plan copies . The original timewtdtcnt will be maintained with the original studyplan . Any deviations from the study plan or from the analytical method as provided will be documented and reporoed promptly to the Sponsor Repmentative. DATA RECORD MZEMG Records to be maintained include the following (as appropriate): Sample tracking sheet(s) Sample receipt records, storage history, and chains ofeustody History andpreparation ofstandards (stock, fortification, calibration) . Description of anymodifications to the method Instrument run sheets, bench-sheets or logs Analytical data tabla All chromatographic and instrumental conditions . Sample extraction and analysis dates A complete listing of study personnel, signatures mid initials . Chronological presentation of all study cornapondcncc Any other documentation necessary for the reconstruction of thc study Cbromatogramg- All chrnataEogtams will contain the following: . Sample identification, injection date, urow or other indication of the area of inicreat, sad injection narnber corresponding to the nm. . Additionally, fortifications will include the amount of analyte added and the sample number of the sample that was fortified. Analytical standard chromatograms will additionally include the concentration (e .g., jlghql.), page 13 oj63 Exygen Research Page 54 of 110 o000W Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: PODU0760 ExyBen Ptvoxot Number: PODDO?80 . As pmt of the documattation the following sheets will be included in each analytical sot : a run shoat listing the aaaApfea to be run in the act. and an instrument conditions sheet describing the instrument type and operating conditians. Ail applicable requirements for reporting of study results as per 40 CFR 792 .185 . QUALITY ASSURANCE The QA Unit of 13xygen Research will inspect the study at intervals adequate to aenure compliance w4th GLP's, and will report the findings of audits to the StudyDiroc.-tror, Exygen M*rlsgemez*, and the Sponsor R. epresentative RETFNTION OF DATA AND ARCIRViNG All hard copy raw data, including, but not limited to, the original chromatograms, worksheets, correspondence, and results shall be included with the data package eubmitted to the Study D. irector These will be archived with the original study plan, amendments, final report, and all pertinent information from the Sponsor. The besting facility shall keep all electronic raw data and any instrurnsnt, equipment, and storage loge for the period of time specified in 40 CFR 792 .195 . An exact copy of the materials submitted to the study director will also be kept at Exygea Research. Exygan will obtain permission ftom the study director before discarding or returning sarnpim Exygen Research Pop! JY of 63 Page 55 of 110 000078 Interim Report #23 --.Analysis of Sediment Samples Exygen Study No . : P0000760 Hxygeo Pratucot Number: P0000760 APPENDIX I ANALYTICAL METHODS V0001780 : "Method of Analysis for the Deteiaurmtion of Parfluormoctanoic Acid (PFDA) in Water by LCJMSIMS" V0001781 : "Method of Analysis for the Determination of Pcrfluoroodanoic Acid (PFDA) in Soil by LCIMSIMS" V0001782: "Method of Analysis for thE Determination of Ptrfluorooctamic Acid (PFOA) in Sediment by LCIMSIMS" V0001783 : "Method of Analysis for the Determination of Perfluorooetanoic Acid (PPA) in Fish and Clams by LGMSlMS" s~ V0001784 : "AMceitdh(oPdFoAf)AnianlVyesgiestfaotriotnhebDyeLteCnInMinSaItMiSon" of Ferfluooooctanoic V0001785 : "Method of Aaalysis for the Detsmnination of Perfluorooctanoic Acid (PFOA) in Small Maumrnsl Liver by LC/MS/MS" V0001786 : "Method of Analysis for the Datarmination of Perfluorooctanoic Acid (PFDA) in Small Mammal Serum by LC/MS/MS" Exygen Research wW 1s of6s Page 55 of 1 I O 0000'79 Interim Report #23 - Analysis of Sediment Samples Eixygen Study No . : P00007b0 Exygen Protooot Number: P0000760 /'' ANALY97CAL METHOD bfaAWNmaba: VOOOi76D MeOo " OfAo*7k1Yq`tY1D1ltrmWo1b00fpet1lYavatsnokAcid (rlOA) k Water by LCIMNpf9 . .:4aAytiral Tetdal Pvility: Pyvaeixaanxh Ma Aaprcb Arive Sua War, PA t"01 App--d ir ?-A c--U. rT..euezc:awriocu.yarr, L.amta, I+hyp-oxe ..vaa I alsao+l Dw I'^+ Exygen Research ~~ 4- 4"1 ~'L k .I Totd Pyr : 7 Pare / colas Page 57 of 110 000080 Interim Report #23 - Analysis of Sediment Samples Exygen Study No. : P0000750 Exygen Protocol Number. PODDD760 DsyvslGwt" A .YALY1YCwL MLTAqD Mbbod O(Analyw0 he The UeoqrmsrWen of LOhtS~d4S 64lkyN,yaenvppOt7lo It Acid (PFOA) in Wuer by 1.0 Scope 7fis mdbod i0 1p be aaptayet tor the belepon podqo0elilatioe of pwfluae00etOpoio add by High Parbtfnuto0 139oW (7e0malepyhy oouPbd to a tandnn Mass lipec4ometr9c Dalectcr (LC443&M in vow. 3~0 8" 2 .1 Allwq-abewvOnlrIObOrelerypracNior . 12 Chapels de sppneipuaxMSDS helhrOhWliq anychmdael Air proper safety P--d-L 3.0 SwnpIhAerriinawt 3.1 At Sam 10 rnL Oftea ample farextrocka 1.7 Hosempliyroow~usieseem ktOMerwnpba )J Sqpplep *M teEiSanted ahould be HIOwOd to equill3rale to roem 3.4 All woplamodbe Utoerullikly adttad LeRn beingswipled farextraction 3.5 Any+tvdpla ceaqiMy PWidwabauW be aeomfNPd as -300D W Ibr-3 minulw andOw wpenIWYAt DhdON dw wrtarsction. 3.6 SORlple cWllehon yiocaduRS will be q10eilled in Ow sYripli6/ plan fm tAit Pwism 4.0 Respowle and bowl" I+ 0.1 Water-RPLCyede 4 .2 ASolhorol-HPLCpede 13 Axasaoium Acalsde-A.CS.1el~mt1 Grsde l0 TaIInoraoslratoicAcid -9IDpa-AD3idl S.0 Io,6Wrmcot and Fqoipmmt 51 A ho .*ermnnnee liqodd e4romWOgepA capable of pump4 op u 7 solvmtta equ47aA with o vsds310 v0lome igeatar enpOble of iujoclin= 5-406 pLevaeeobd 10 e hndOm Mw Splehnuur ([.QA6yJMS) . 7.2 A derlea ro0oU0el mw Atu Ilepask iateyadou and qu0mituioa 5.3 Aaely6salLnlaarscapONestreadnKtoO .Dq001 ll~ 5.4 SO eI. diyowSbpolyxapyleue exaaeih.Se roba . !.s ISm2.dlepemYlepa13ya0pylntec,urriifa0,bbea !.5 Dimponblemfipapbpos(lG)00u1, f00d00ui,). 7.7 173iaL LIRBatucowaaaehDarJn. SE 7mLolurFiP'LCrWkk. !.9 biepoubY plpmllez. 7.30 Autopdpetla (100" 10p0 pL ted 1.0100 rL), with dispoe" tips. S1! Waters Oep?atYae6oa(1t) ICIiEPEtYrnidpa. v,e. : ..r+ . Pat" 17 of63 Exygen Research Page 58 of 110 000081 Interim Report #23 - Analysis of Sediment Samples p. 59 EXygen Study No.: P0000760 Exygsn Protoeol Nmnber: POOD0760 r.r,ws.. . .Rs brdhoa orAn.ly~ u."dKre. .v0eonlo A11ALY77CAL hIL77IOD lhr,wovremlo.doe orFatnwMoounols Ae1a cProN in WON by una"a 112 BDa ..ewum.elrew. s.is p~ayra~,tf,ar~i~50ar,VOVypropym= n,a..cwo6rm. 6.0 CwMawr"We sy.M a.l Arayat.l oa,m,n:.plwyew (PrN:uwsuu]3o> RP nc.ymaw ftnm6ch x.i mm x So rm. 5v a2 Tamp4uaM )VC 6.3 LiobiblMre (Al : 2 ol0liamoeitim Acedro 1a Wtx 6.4 MshiloPhnr(B) : A6t9Met 6.S Gmtim# hop= ISAi.401C1 YsA 03 65 lA 65 S.0 25 20.0 23 27 .5 43 Flow Rus 45.8 SmLhmjy) 35 0.3 35 0.3 71 0.3 75 a.3 35 0.3 6.6 IOJootbu VOtYme: IS pL [wpbe inccsued ro u mxcT a30 pC). 6,7 QuLntihlioo: ~edcAra-cximul~t~a6adalibrariancuwe. 6.i Rws15mc -23 miNnx The a6mr<0edltipsl an mtoaded u a Sddr ud mRyDs4]umgpt b, ader to opda~e thr I1PY.C ryveam . 7,4 M$fM6 33atm 7.1 tdOds: 8lecpuWnY N9D6+ti )rlRhi mOd$ . mmdhnin; 433 - 369 mh. The WOYS ooadGiON Yo 3oteoded r L yuide md WAY be Ctnmled m adv iC~ aptimisp.lhe MSAL4 sYjqan, I'D vv=rloreaU"w 1 .1 "it Phua l.l.l T ~pli ~wpooimn ~ce0~9s in w~Ox ia pcepa~'ed by a0dinl 4.136 Y o! wmnplnnweOU b ]OWml.Odmiw. A1SaaW voMwm may be plepma6. hysea~= PdBs !8 Q1'63 Exygen Research I~' aoi Page 59 of 110 000082 Interim Report #23 - Analysis of Sediment Samples Exygen Study No . : P0000760 r~ Bxygon Protocul Number: POODD7b0 6~qpplura4 I/npYd N=be VYp017jQ ANALYTICAL MZTHOD b(lihsd aiAotyeie rirlks Ds~nOiw afPKiluorooclan6ic Add (PFOA~ in Wam by 1CMSMS 9.0 Studied 9.1 Steadied SleeYlAertillation Boluiau 9_ ;_t rrqw .r.~ .ewaoa or-1 aa Iy~ILor~+aA b~ir~~ Io ~,,~ o< asel)oer+ha .,a(=Wed fu rxity) wa 4kh1cto 100 mL with msOteol in n 125-mi LM bokb. 41s A to Pymi. somne+aon 90mion orProA i. yrepl6a by brin0ie010 MI, or go 100 XWmL solotion b s and Volume of 100 w i14 ~nsswnw is s 125 neL LpPB bapls. 9.1 .7 A 1.0 *BhuL lbnitketian apimivp ofPPOA is pnpued by brinjing r~i ML of1At 10 K&6oL wlwNon to o flew vubam of 100 with ~ihuwl io s 123 mL LOPE bottle. 9.1." A0.1 p#~nL fow46ceuoo wlution of PFOA is prqutxd by briayny 1 1) aLar>5o 1 .a,tyiqL 7oluo6. Do .ft)vOIWna of l 0o wilh mNhwel in 017S s.I. LDPBbokb. f.l.l A 00.01 kvsL meusoelioo soAroon orpppA Is prnwW by bda0ipk lD aL or lba 0.1 W+mL o6hRioo to a (but vWLUM of 10D wok dAuml ins 19S mL LDPS botda 9.1.0 TSoAadred rerti9osdon solwions seabbe Owed in srolS6prseor et IppRKIptWCly <'C and aresqble fog smsximum Period of 6awrnhs 6&m Yr ago afpnpcpior. 9.2 0tandse0 CGtifw4oa Solutions 9.2.1 LOIMlIAIS talhrnfo6 nuufuds in pq~sasd in HFLC water. The odiYwdOY fluldards we peoceusd tArW/h tho ssnaction faocedwe. idainiodlo nmpLs, 4.2,I 7bs 1411049 is a Sypdqi menplec IdOfdpol ooneeatratis6ss msy be pepns! a. needodL P (kaneitrsmu oIPOpU9opnor 5610105"b) 0 1I0a Itooo Pbdlfauon VeLmu o1 Vo1oaee PanIEd CbnWnl 0 lro 40 40 :66 +o a60 48 lob +o Cammaftiou of CaliLstipy 3usdred iDs 50 l66 zw tYllhrnioe Spuxwd II7 -e k X xcmffdayy.l Xcb,nayy.a xxQareor,.dadayyyy.sa IOD 200 411 006 4 S06 XClurWdyy-0 1666 7CQa~eddf7rfi ~Tm s~oolua aooomm.Iioo orms raliAnnooalMend is pusu m te ;u alidu oopaeslnNoo, dso is rM wooawmou ohbat+nderd dwioB the CRUMi0a WE). xC-ceapsd'nlibrsuon xsei .ea. py6i W" J01- Page 19 oj3J Exygen Research . Page 50 of 110 00~'$'~ Interim Report #23- Analysis of Sediment Samples Exygen Study No . : P0000760 Exygen Prol0eol Number. POqpp76q &yrrlw0r .aa waedxwj .evooalrw ANALYTWAi MnKM ld~od oTAoqyuie tbrur Uearme~em urrerlWmoeeumiic Acid tPa ILOIRMS rp in Water by 9,2.7 A Um aenlaid eolecin (ejpeq bta*j mma bo unmat4d with ooch at ol,rpedrrde wdneled. 92.4 Blon e!l exireoted nh'haSiea Meedvdr in 1 S-ML yvlff"Avw tub" r 2'C lo6'C, vp to tom waka. Y2s Atlaae7s voitmr ad oaroemruiooe of amdude may be pop" n MWIL 19.0 iiekD Ss Uf Ial 'ZKA YYaA or emp7s e=eeLwO (e3pfeldly 20 or list) mue ioeludr n Icuc on seOVA onlrol (alalbod bboit umn HlyLt wow) rd tvro m0ont omeodo fttillMd or boon ooomntretion (kb cannot IpRe) to ntwly Re tbs back. veriApocd" I6.2 Raqodnonoatr far Gold said Iobeneory dvpilalae end Irikea will Ee speci0od in to qudity Yeurmoe Y1m Far die plojwe. 11.0 SmorpleEicheetios t .(.1 lduema 40 su.of AMeepb K e yeruoo ut UNDple dilute6lo 40 ML wiEa rates ime $0 ML 1olywovrlene oeoa;mje a16ed (Miy AM eeedvl, replee 5d IM ma win 11 .2 Condlioo tbr C>, SYE eervldpe (1 p, 6 ML) by PaWng 10 ML methanol tollowadby S mL o(HP1.C valor(- .2 droyfnoc). Do mw column mndty 11.3 l.aed wlpAm wpdipood Cu 8f8 awlridpe, l~ecerd eNatr. 1011, 11 ." MOW wLb -! MIL 10% mel5rwl. Calbct 5 sod. or duet, into grnfuaood IS 1p1.Aotypopylae oestrlll,de ro6ee (6me1 whm,e - S ml.}. 1 L.S Mob- eemplw ucqJ aeelroeprtY LCdMSlMS . 12A qmose"repity 12.1 12.2 123 lZ.! bWortlAe ewe otwmd of Mb fAWda:d, Sample epd WOON sample inlo 1he LCAIHr1dB spun A eeoleatlen pmdard mw praoede WA follow all ae.lY+sd umqa SSandudraf9B0A camAOOdln~ ~ et laat iirv or mon oonoanuetian levals moMbe =WWin mroatlticolae4 An Elfin art of ractraelad ahbndor qaabrdo W" I* !aglow v Or blinaio6 ead at 1M a+d v[ e sample ret_ Extracted rtordm ;de mug he Iptrmpirsed bdrNie ovary f-10 aemplee. As on ilteantiw, Am entira re OF 9xtufed cribraHoo atuWrde ntay be isyecead at Ilsa bagirm~ of a 9e1 16dowad by oaRSded embistloq uanEode inlerrpaaW every 5-10 fea+plel (to room Me a retOmi in of exveeted atiot0aeda). in aimflr em, exvacted calibration pudmdsmoat be ft Dnt Pod JntInjection in asample ect, Ue0 Now dmdrd cwva Lor qwmhwim+. Linero sand" cumn at preqew(Udw adYtaby RawroVewiaa neiqs 1h weitLting of pcel a,ow v.f. s en /^ Pqgc?d of6f Exygen Research Page 61 of 110 OOOON4 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P00007fi0 /"" Bxygat Protocol Nvmber. F0000760 ft1r .2uuP1 " ;et" Npw&ee vpdp17lb AP41LYPfCJ1lr MEl'SOD " bteiiod of Antlpie forlboDcommlaYk~~of~c Acid MA) ix Wata by whpe oNl'bcadm "md~rQ emo~lYYiou udoQ Jsdrs4ymc 3.3 (au equivakNJ wiMan syYm 129 Ba4pls aelpoans aboci:d not aaacd at~odod rs~opcp ;y,y sc111D1e: Ihx axceed auod~d eelqcaKS Aouldbo fw0lad;lated low nuulyaed_ 13.0 Aocpnccepitc~la 13 .1 {:bromsupsn went diner a PccY at* dLC*ow ioa ac 361s wlr 4clo a petan of 413 ML TSMM 413 am prsse cornqwode to the PPDA rnioq while The hp6111ecial(569 wu) ~+R~Is the ba u[earbcndiceide. 13J lp9ud bLl11d anllt aot ooafaioD ?FOA st Jonle pnclr than tbe L(JQ, if a bYet Contains FFOA at IwW prlr Ow SO ciipfli, Wn n now blank sunDJe rpaatbe abldmod aod do codes set mist be w.mctaeae0 . i3.3 Recoveries of ooesol *km at roatrint aPllus ram be berarm 78"10% or 91,11r, mowa tietua Us,aepollDikr hibolrtodetlW aoepfableIlmlls. The native so of rp]r jneL 1e Isaatuceed. Any~mAia Ipika enlnide 7.0 130% sboald be ovdnate6 by ibe aNlyet 1o detwnliee if ee.atlnctio is rcRaudW. 13.4 Aqy alibndaa Ysodnd bud le be a slefiefieal we0iae by otirl6 dle Hao Enoe TM way be axcWdOd Ilaln dc eetanlLlioo of ft cdibratlon cavrc. 1lee+wa. Ow topf rmabee of oa7wK*ed p2ibp0tel Ousludo Ow could km ao0r11d meet not eLwed 20% of Me low vbec of anew elsndardc irycowt 0.5 7fa sonn]cuon coc$isiek (R) is t,allbration ouvo ylalasnlcd most ba 2 .90 A2 iR' x .9150 ), If alibemon me* AII oubids Ilues 6min, d., be WCOR M ~ and Ihe ~ ~ srWOt ad odat~laa choo r~ptnlY>~ 13 .6 Febsldoo these bdwun clnndnett sad almfPha must am MR mm then :4 %xi0linae wllylier nm. veebetioe tlmr ddRexceeda ths iSmilwilhin an andydcdrue then IbrmmiMDe nleoclynd. 14.0 GicolNlolu 11.1 Uw eAe i411owlns poarim b eafcnWi Ihe I~wum of FFY7A (ow~d tin ngrL, bud oo Vale uw) usinQ tbe Mndsrd eeeva (linsllr reFueioo Dsrxaitunl aeiltcWdbytlr VAN L,ynx wRwwc ploree: PFQA fvoed OWL) - [Pd( seen - inusneoe x AF +bp DP - hawby whieb do Bonl vdsma ww dllolad, if neoewey. rue6ad 7 /"" Asps 21 of 63 Exygen Research Page 62 of 110 dD 1111" Interim Report #23 - Analysis of Sediment Samples p. 63 Exygen Study No. : P0000760 Faygen Protocol Number P0000760 00- eqpalreeucr ~r.maN~.. vooo~o j ANALYTICAL AQTHQD ~~ for7u15t1rmio~tiwafPslloaroooWOkAaid(pFOA)inWOtaby LGI$91M3 1l1 thrsreplLe mtXfd with loroxn aemamb of PlpA pdor to exnceFOn. use iM IB71Ww'loj equmoo loodaohlo 1M ppent :ocovay . AmwarD (%) - [toWrilrtlEtsd Gyl,).' amlyb toutdia camad(u~Ll] Ewlyle ~adea(n9L) - 10D rs Exygen Research . . np7or Pagt as ofss Page 63 of 110 \~\/86 OoOl) Interim Report #f23 - Analysis of Sediment Samples Exygen Study No.: FD0076Q Dcypn rroloool Nwrb: PoM760 ANALYIiCAL 31fETHOD bletLod Nwnbe : V0001761 mdLOd a1AFdp(f ha 1b5 ftMmWadoo afftrAuwoeetaisalo Acid (rFOA) m Son er LCIIASIAti Adly6cd 7rbpP Facidty: Hcp= A~b 3Q38 RaeaaA Dri+'s MM COW PA 16101 Approrad SK '~ L.J1J~ Paul L1inmly T.olnaeW :.r.aw,r.c .bas,axrnuxaarsb ~jo,j,tj,104 Drts m l~y v~ 00-1 r,&~Lt naw Exygen Research ToatP+pa 7 . Pago 23 of 65 Page 64 of 110 000087 Interim Report #23 - .Analysis of Sediment Samples p. 65 Exygen Study No . : POOdd760 sxnel, ranllwal rrwntwr poWso aa,ftie."A .,.MdN..,,.,OwiM AxALYncAt.laereoa MAW ofA7Wyait 6w IM Dauemipatipm plPwfIuWOOCUUofe Aaid (PFOA) in Roil by 1.G2H8lMB 1.0 Scwpa 7aii malbod iu On be ~plvyad fm 16a i~eWioa aM qwotie~tiq~ olpaAaavatanac acid by High Iaibooaoa Liqldd CkwnlWaPby coupW to a qnqaa Muc S~sehomatric Odadar {~.CJ6(SIM8) ia soiL 2.0 fa8lty 2.1 Alvap aboars ab bDoeraqpraehcs . 2 .2 (3bmmItiLrappmfaialcUlDBbofmshaadfiq8alyoApmk,tioipwpsraaicy PrwmWwm 10 SaVbRpuinaed 3.1 At 2ert1l8ofla2aaNPb16rWCVaccori 3.2 No wwnple Proerlies it naedad tr wit ampla. 3.1 3mylr a0oeodrohf,llaled aonld ba aliovrod lo aquiilbraa to mom 3.s All aompid mw be YmrouWy mixed Lafaee beint pmpla6 fa mcpaoGon . 3.S 9UoiPlo f9Uwlao pvcadmsw will be ayecide6 in do iampliuy Ptaa PM On 1~M- 4.9 Eap4ir ad &raRam 4.1 wow-MLCg.dr 41 Llwhmol-11PLCgsde 4.3 Amnoadum MOO-A.CB. Raaynr Orndo ,C4 pfffhwoaroloAcia-sism..Ala;en 5,0 lnmument ana Hquipmeal S,1 A hljb ymfUmaooe ]iqu16 tWoerROVapp eapa6la of pwwpin8 ~p ro 2 mMema eqmpped aitb a ruIa31e volaxar ioJaetar aVaLle of iajacdn8 S-200 pL aamaoud 19 a OW= Mm EpxYOppta [LCJMSfMS) 5.2 AdaWotSOS0UeoIrawdmltkPeak lr,ta<rntlw,andqwnQulion S.3 Aaalydea1 halaate eqabk ofiodin8 to 6.000018, $.i SD mL dijyeabls PaWoPylrie cenl6U4ye twbea S.! 15 ml. Aiqp0oblr Polyprepylsr aennifoge tobs. 116 Dwowbb mksopipep (304100u" 1004200uL} 3.7 12S-mLl.DPH asKOw-nowk bqtla. 5.8 2 ML elw nPLC list d.l 3.9 DiayaaWopipnua S.10 Aulepepa0r (1D0-1000 pL ud 14100 pL~ wilb ~SpouEU 5pa . 5.15 Wa[as 8W Pak Vu 6 a 431) SO " SPB caruidyp. 5.12 OBraeumouqito1d 5,13 UlOarooiebWIK PVs2af) I~ " Page 24 of G Exygen Research Page 65 of 110 000088 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P04QD7b0 l:hcygen Promcol Nuenber: DQppp760 saf7K AwwONf itauot en-, . v6o6tya ANdl.17TCAL AlSTii4D 1~IethOd otAwly~ek fw tLeDmamiorioo ofY~tlluoroocwwic Acid (PFOA) M Soil try . GCIMSII6S l.14 SVebLaWm eLdtw. 5~1$ cA;ne%PobearqivAiw %Q mL Pw,pp,im, ttb .w soaQ rpm 6.4 CM-M*Vhk sY.w. 6.1 . AoJytiod Cdnar, FWephreRP (Xeyeleme$pinfjfir), 2.1 MM x $0 mm . Sp ErfN: 12707-032i30J 62 7YmFaapw: ]0'C 6.3 Mobile Plww (A) : 2 mM Aetewnium AcaYts m Wptr 6.4 MObiuPbwe (D) e MeD4mi 6.5 GtrlIaotPmgM= I1Dato{Dw 1i~sd la 6s ea 2$ 2a0 25 2T.3 61 PIOw Rile 7fH 3s 1Dia1.p3 IN 3s 0 .3 7s 0.3 7S 09 3S 0.3 6.6 io6CIi0qValue: l3 pL (em be yq6P06606 tn 6e mkh p y0 ytL). 6.7 QueWIaYiop, PpkAna-mctMwlaodtd=fib,atiaapurs 6.9 RMTime: - 23 0ft". 'Ibaebovs oomd;tian w i~~e6ed w a lpw4 ~d mry bs eh4a~d in mdp to Optlmil:e do 117W eyMm. 7 .0 MS/NS Sysam , 7.l MD6r EUe0wp~4y Neptive ABWrI pwd6 monilooin6 413 -~ Jb9 m2 for PFOA . 7!e ebsw taeditioee 4n int+oded r a juide >Jrd my ba cAr*ed iLS orda to apqmiyr to 1lSll4 ayWem. 6Q Mtp4WawMSsL6ion~ a.l btob3le Pbw 6.1 .1 2 m8[.wunamlm aaehte in .rrn 1, peqvd by addiay O.IS4 got sutmaoiom uaqlv to tUDO mL of watcr. AMdnete rorlmnr wq be pepxd. PV I ory Page ?J of 65 Exygen Research Page bb of 110 000089 Interim Report #23 - Analysis of Sediment Samples p. 67 Exygen Study No. : P00007G0 r PaygenProtocol Nl>mbo.: Pooo0760 , . . +W - " bUehiN" wM v0001'rJt APULYI7CAL h1E'1'HOD Mabod of Aodyefa Arr tLsDeowmioseaqof PsBnamotepeio A. cid (PFOA) ia 9.15y - - 1,C1M.4+M8 9 .0 SWAMRqrariw 9.1 ssodapdlbeifFeedBsaliep9e/ulios 9 .1.1 Prepo a Aod IoLltiomof -tOD pjlml. afPPOA by wei`hiaj l0 mg 0( ~ +~edod (aomaled !or Pmity) nr1 dihoo so 100 ml- xdh selbissol ins Us-ml. IDPB boet3r. 9.1.2 A 1QpymL ~dcnion >taiwiw efPlpA h pwPwd by br1r:)eln! 10 anL ol1Le l00 Pymt wlution fo a lkal whme of l00 wilb nxW~nol in a 125 feL IDPE bonle. 11,13 A 1 .0 P#m1. krdSntiom avlutlat olPPOA is psPowd by 6vinfing 10 n1.e! t6e l01101IeL talotimn to a liwl woLane o! 100 wilh nxdr~el in R 125 UL [DPH berqe. PA 4 A 0.1 prmb 1brtlSatlm adyqoa cfPPOA ie ;upuad bylsiyins 10 ' mLaillAe 1 .0 P6hni.aebtioa to a MW volume of 100 with meduewl in s12stoL l.nrs ln,lee. 9.1 .7 A 6.Oi pV'n,L fotitimtiom nlullm of "OA ii prepwd by brinyinE 10 imL of Ow RI pahsl. 70hWa1 1o a fins] voinat Or too with nnqeri la a 12i mL [DP8 bMQa . 9.1 .6 TV Mock endftetilWpon solstooe sas to be Itond in ambigoratow et qProxhw0ety4'C sad mIpbk Ibra atudnstua Poiod of 6 monOr howtbado*of pnPoeetia~n. 9.2 fnndudCehLatien$nlpdem 9J.1 1.GRdSRdi aUbraqon mdtudo :e ptqwpl Ia HPLC wap. The ab'brt$ao wordmde are praewoed th~ to ealsacliou proecdurc . idenDlal w alnlplw. 9.t3 711e 1alowi .y is a lypial wnWle ; a0d'Itiwnl oaacanruionf may be pnaaad Y me" Finel Gbaolnbboa PoqIfullioa VoRrmeef CbnoaiYarioaof Gltibration efFaAifioalien Ve1ms ia1i11edCor,v1 CefibnNoe StrdrdlD Solmioefodd (PL) SunolelnLl Standard looll " [wMo6S 0 o ro 0 xanmadyr~b " 199 49 23 xcmmd6y1~1 1l0a a96 ao $0 xCMrmoyya It 400 40 IN xcinmdayrl IDo lao 49 sw xanmddyl4 lao sou +o so xcinn~ry-s 100 ~60 ~0 1600 XCmeddrr.e " 4lwooctrauroei oenomaulos olSlrullbeMios Wadad it aqlrl m ix 1U inilif,l ooaoaqnqelt, doe GO do eoeomtretfop oftheetaodard daAn; the extnetion (SPE) xc-exvaere railbcvlon Itandeed . r4 .r+ page 26 Of63 4`1 j Exygen Research Page 67 of 110 000090 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 filcygen Protooo) Numbor: P0000760 ell aww itexame. Mwoe Numb" voonn6 r AmLY7ICAL 61Z7HOD ~-6LlhoiafAadyw Jim be Ditamioedop of Polfiverawas(o ,1e{d (PFOA) io Sail by 1CMBM3 91.3 A ~ akmdrd moLtdco (rea6eor bVSlt) MKIellmollisd with omch vast ePauederdl prism" MA . MM a4 otraeLli crlha4np aadarde in 1!-mL PelYpnpY)errc wbcs eE 2*C in 41T. wile to two weeks. Ahwnohrolunao rqi ooperahrtlar atawdds maybe prepmed as feaiYd IOA =eleLlpe[Ip 30.1 19cH Wock of orarp/eo arliaMe/ (1)yipiHy 20 or low) Coast hisclimle, or least on lWlrrl cosehol [matlod bburlt uehrt 5 sell. orwtbaaei) mod two ttveor eoohoU IbtlEad at known soooortrWom (kb eorrno) epSke) m verily pnadrY nawwY fortMl~rh. 02 AeOdreemoolle ror SoW erd lebvn0ory dotliataa mad eplYa wil l be specified iu Or9~h "" """"""" P~ 10e tLfr pqjott . It-0 8a"plb~rkreliw 11 .! We(ye S S or nmplo imo 10 mL pobpropylana cmdftlrya ntbn (Aedll" u No" "Place lid nd wax wow us Add $ mL oraoelb.m] .nd dab m . Wrist aouem lbererra -I s aluirsei 11 .3 7naafir, flee a*er is au ultrawoic barb end oooicaoo Ar -15 miavks. 11 .1 Brine the rqkuoe 19 m 40 mL with walur in ft SO IoL polypropylene orutri0iyr lua. U .b CermillqeFor-lDmipohuet-)00prpea 11 .6 Can6i6a11 tLe Cu SPH cuuidget (1 s. 6 mL) by pauiey io mL mnhenol ibDarved ey S m1. orrF1.C rvetar (-= dropMo) . Do not let columa n+d ary 11 .7 Load (deeoaO Ox =MW on dro caodiuonad CGr SPE eantidst . Discard eLruu. ]l .t Blutr wlrh -$ mL 100% welhrml. CoUoot $ ml o1 oluue IMo priWwxd IS mLpo)yWupyEma omaillyo tuba fEyieivotume-! rnU. t1 .9 Amtyioatay+laiWoeeloeeroqerayLGbttJldS . la.o aaammilarallity ml how am etaammouet oraKh iordmd, wmpb rd 1hrU8od tenniio Ime ft )LCIAI6IAES ,yellies. A eahbeulea oloodard end preoede end fonow all aomlyaad uoplm. 922 SaWr* alPiOA emulrleelo&a is, oorrnpw6bj 40 sea muyae.l w. at UM Ave of SAM MMUNUM 10"k 12.3 An saden :x ot eomrcrod oaft+don qlrfOmda mot be included u die begliessisrj rd at Yr ad or a eunpla at BzUSatW wndorde rowu be wknyowlelrrwn every sao reasons. u an alomiw4w. me ere en of tW $ .re 001, - AVe 27 oJ63 Exygen Research Page 68 of 110 0000'~1 Interim Report #23 -Analysis of Sediment Samples p. 69 Exygen Study No. : P0000760 Haygtm Protoool Nnmber: P0000760 'I)qVn4usdr- . . . Method NN*K V401781 L APlALYttCAL MSTHOD MeYWSd of Aoa>yw f~tba Deorevtertbn ofPnfluaoommda Aeid {PPWy in Soil by LCIklEltd9 aaqKUS alibatlen nrdlmdr may be blJeaad ilt do balplnabt= of r, ax foRowql by a4siad aitMradw tlrfdvda ioledtpaaed anry 5.10 sumples (ro ON= Rr tl uoand sat of avacpd rundwde), la Htlha cae. at,tfacud . ollblraFpl ataedtrde mum LrfLa hat aad bee iojaelian in a nmpk teL 12.4 Use law aaadud serves, 10 qurl~eKim, Lisar psadrd serves ne pauntet fOr Ur snlybblr liowrnPveou rrfaP Ux vrofFAt61` oFpe+t araa tvmuu oaRb1111oo Ipaldpd aonoenmtbn uft MrpLyra 3.3 (or equivqtet I soRlwrayWfe. 725 loirvlo rapsaa rhwld sat aoceaad wndtad f9foeeua Any gp*k* that exewd pmdmd=erpofreq eb+uld be Audw diluted and reanalyzed . 13A Aoorptanoo CliOaria 13.1 CMomaeo~am mun show a poak o(a daaPhfor ion at 369 amu lhom a puem of "11 wu. The "13 oeu yaraut oeerwporbs to tlts PFDA stim while the OauybM iau (369 mu) mpaaqa iLe low oCcacboa diuxid . 13 .2 Method blalb mint not wnaiwPFOA ri Lewis arsdor thsu the LOQ. 1f a b" eo.efNW PPpA nlrvw jmaurUM 30 ejr1 . thin a oaW blank sample mowpeobabwd aodas mtlra art mustloo ro-roam". ' 133 Reoovniea of eondal spiltp ad m~tna spilna must be bqwot:a M 130% of link lspwa valaaf. 1f a control apr'Icw folio outside do aoowxable ]imua, the ' ap8M wt of uoiplw Woald be rfaxtnced. Any autuix tpiF.e outside 70 IM should bo avaltatod by Ow twlyrft to dolamiue if fPf7[Q74IImt if wifYdid 13-4 _ 13.5 13.6 Amy cabfbimCC ahadYd bold to be a ltYilliesl o111Rt1r by utbq the Huge Pnor Tar, row be eteludod 2m dr tdwlxian of tltr eaRbrntion turve . llawvu, 1ke Wag euubar of ectraodd calibration. stood" that could be oxsluded mmt set axertd 30% of do WW mlmber of extracted stand"s We" The oonelatiaa ceaRStasd Qt) Ix aiib,atiuol outvet Pemarated must be iD.9V2 [H= 2Q9R5} It oab7ritim results fall outside these Iimur, thm approprHaa slop tnwl be tntm to ad/utt inttnomant operation, VA 1hC Wodafdrnr 1herekvuat aetafampko ~wttdbe rvanalyaed . Aatenlfoti times baqreep moldtda tmd samples mw not MR more dun *4!4 ritphaa rW~dultav<. 1lfeteation tifsa AriR aoorW11da Lzo1t uithin bltYWl'h~1YGtbp1lh(frqlRUathtr7YN1y7Cd . ry.#et7 Peps ?8 oj65 ,a ; ; , Exygen Research Page 69 of 110 000092 Interim Report #f23 - Analysis of Sediment Samples Exygen Study No . : PQ004760 I"` Exygen Pmmcol Number: ?0000760 :winft ""Rb r,wAee 1u,anvooorni ANAI.Yl7CA4 AlLI80b LiwIAodofAmlyis tx do Dqaurtian ofPmfluaroomnoio Acid (PFOA) in Soil by I.CIM9DdB KO Ca6ar1+ve4 1" ,1 Usa dW lDHwr*oQariloo to wlpd&0e 1bc moum orFFDA Forod (in ng1L bad oo yp4 nws) -in Ih. e-6w - 0kn- reFaWoa F(rWAonl inafaa+b tLelLU W- u2wnre Pmfm: PFOA fo~md (ol+L}~ ~t ana . feouaeet1 x DF WOP DP . Jimoorby w6iek Ilr aod volono,rr d1med, if naaepnry. 14.2 Foruqis Andlld wlA Uaow mumof PFOA prior to caurclion. we the b0owiog sqn.4oa wa .liuberm. Pffonq rooorny. Aeeo+a+9 (%)' [toldaL!h*h.d(,aL) - ~1~~ ~ ~aorot{ay[.1] ,100 -kto nddrd Gy+f.7 14.3 UM d~s fotbwio6 equdioo lo convert On arnarmt af FFDA [hund in tulL w pWLUPb~ PFOA hod(ppb) -lPFOAIbaod (ndL) t yoYgUe eUacied(Q09L11 ompkwa*(S j) 14.4 Ux do hAvr.ig oqua<lon ro erwrax ft imam or PFOA bua in Ppb brWan ay wajha. PFQA fowd (PPb) dq wai;Ler FFDA fsud (ppb) x [100% ! qW yolidaf%)1 rys ?era /"w poge 2 9 of 65 Exygen Research Page 74 of 110 O Interim Report #23 -Analysis of Sediment Santples p. 71 Exygen Study No.: P00007b0 Sxyyea Protocol Number. P000076D r% ANALYTICAL METHOD lA*0od Nwqb~r. V0001782 afotlhod of Ax*nt4 Sor As DatauYMov, of lr0anropeyapk wefd (PPOA) 7w tlodlmeatlyLCl61l14MJ! - AsWytiet111tOtinSFilitY EnYpwRwweh ]OSi Reeauch Ddvo SIw Collpr, PA M01 AMowdH7 2=~ (_ 11, PAW Comnur Tab" LeaOa.LGdfs~ 8xlpo Raewh igh61,104 Dut9 ^ Exygen Research Tow rosa: dr Pogo 30 of 65 Page 71 of 110 000094 Interim Report #f23 -- Analysis of Sediment Samples Exygen Study No .: P00007b0 ExygeA pmroaoi Number : Poo007ao 100- Iran RqnlH Wafted NWOW V004 702 .urAt .YttCAl.MZTMop Mtlhod ofAeoly* hr Ow A~mia~tim ofY~lluwooounoia Avid (PP]A) in Sodimpa by LGWSTIS 1.0 $00" iW molbod ib babe mploya lbe Op iw>wdae md awnliution orpaAisorooctrwic Mid !y Hiok Pat- LM" Cgommonoy ooupled to a Andan mass 8per-eaah7a Dclaalor (LCJId$AdMim ~odfiowf. 2.0 9~Aoq~ 2.1 Ahvsys aErRVS ofi LLaveoryp:acGees 2.1 CunYdt Ow wpaD4ate kSDB bda+ bmdNnj any checdcat iar mnPR ..Cny pkwtiaK 3.0 Sumb xquirazaal ~ 3 .1 At iwtlOjotwnsaap1o10rviaKUae . 3 .3 No wuploproowdpjlbaeedWlbroo4mratamplu . 3 .3 Drapla NOW nddler+lld should be allowed w oquilibrua to roam 3 .4 AOwap)amwtbtlhaouyNyWeed beAoobdajarmplrdforexsnclinn . $ .1 Swnpk eolkeuon prooodww will bc Wod6vd is Ow tiuapline phn fur this ptalxt 4.0 RaSenb *ed Somd~rdt 4.1 W_Qa-HPLClpadr " 3 AvedcAvidrAuSatrr9k 4A AmmonivmA0elde-~1C.9.1byneMade 0.} POfloomoehsuieAd4-SiprAlddelt 5.0 pql[uanmaadBquipnN4 S.1 A bi# pwf6anaiw 6qodd CpOm-WVh tupbls of pumpiua up to 2 yotreap eqpiVpad +W4 " rrehblo rnlm iWopar Capable ofinjecURS 5-200 YLaon~po~dlo ~t~ 1YIw Bp~eav~~NrSLClMSfIdS1. 32 A dwia to edUWq envdaft ibr peek iaEeSrmao and V=Wutim J Awlydczibal~ tapeMe ofzudbW to UM1 1. 5A $0 ML di4po"10 pabpro"10s, Omlftp b1baL J I$ ML d1sposable, p4pop7ltns castriNee Uibm :.6 =I . W-2. (so-, OX '00-MO-L). SJ 125-MAL LDM KKMW40~ bWa. M 2 mL ckpor )HPLC YW kiL 5.0 Dimp")v 3 4 ", $30 AW9410K (MJOOQ 6LL MA 10- 100 PLI VACh dkpcubW 3.n Wan 99p Pak Vic 6 oc (12) C II SPI curl4ps. 5al OFE vis=oe MOILIAL P-10 I Id I Page 31 Of6s Exygen Research Page 72, of 110 000095 Interim Report #23 --Analysis of Sediment Samples Exygen Study No.: pOd40764 Hxygeo Protocol Nmnber' P0000760 atyW Waert. j MOW cfAirl a,uoarx.e, . voroi?p Arr.u.ttrxst, eMSOa hr dw Deoenniwlw olPuAnmooanmic Acid (PPOA) in SsdimcM by LC1M9" s.l3 von.~r. s.14 wria. .aioe,Ldoo,. S.li f-hi5Pcaqbl0oPmiceini SO mL O*4S"S'kmwm .t 3000 qa. 6.0 Q~eowYo~PYieSy~lm~ 6 .1 AmqroW Co4ioux Pluupavss itP (]fay2leae 5ciawisc), 2 .t mm t SO mas, SN (PIH: ~2SOSC32130j 6Z T+qxrwroe 30'C 43 btabibFliw(A.) : 2aWAmmadumJleruWinWmtr 6.4 MobiUh6rs48) : Whim) 6.6 t]ladime bo0m ; Flow Raft 0.0 65 35 0.3 I:JAD 63 33 0.3 23 73 0-1 2.00 27 73 0.3 72S 6f 37 0.3 6,i Ljsi=YoI~o~: 73 KI. (c~u 5s imw~edlo w much w SQ pL). 6.7 QwoduBmc Pmt Arr-ememdoWAtwq WlbntMdeurvi. 4.1 AMTSma: -13=WML 71t~Yov~ ooAditieoaxn iotmded u a i~o md m~7 4e chiwY~ in ar4r W opdam do FK)LC &ysbnL 7~0 il8Ad38yrko 7.1 lfodc 81wtr62ywy NqQvs MM awdih mcoitadni 413 -+ 9l9mrx Ow PFO& IM rbovi *=Ad= AM rotetw ar a 0" =W may b . dtnpd in mdor to oI , , to MSWB Iyoan l.0 1hepvn6oabffaWtioU 8.1 awoea.re.. a .l.l i oac mmmot6m waa. in run u ynQ.na hy .aaieS QA s4 90f ra~e~anieu wtw ro toOQ w2. .rv . .as. nc0 ~~e r ell page 32 ofas ~:s~<M~~t~ Exygen Research Page 73 of 110 000096 Interim Report #23 - Analysis of Sediment Samples Exygen Study No . : P0000760 Exygen Pr+olocol Number. P00pp76p Fxflr"2"-wa alaaer Nub. vaso 1 711 Al"YTJCAL MAlOD AfnfodofMaqhdt bt01a RFIameeWan ofTadluaaooctanok Aeid (APpA) in 9edimeet by LC~I8IAI8 81 DIoaRloe Qo7wlala $.7.1 18 rada aaiJ p +171oc ia pcapnad by addind 10 IqL o! acedc acid a IOODmI. eftiralw. Abaeeak +o1alFesmsr be pwpKrd 9A il+edndTnpeaalkr 9.5 1Aadud 9rebFavd&aNOa Salaean 9~1 .1 lrMan a aw3 odoboe OS-100 plohnL Of C+FpA by w"ieAinl WiinngK af rruuyekai ateadaud (smrooto016e purity) aad Mule la 190 mL wiih vielbmoi iiL a 12f~m1. IM batels. 9.12 A 101cymL fortiljealOn 1FInEka of TBOA ia pnpmrad by bringing it) mL of do 100 ponL aohmiva to a SW vahono of 100 wi0i mMosa 'so OtS mL1.PP8 boale. 9.1,3 A 1.0 K&L 6orliAaal;vn ool9doeo1'PPDA is paFand by bringing I D ML of do 10 pymL aOhltiOp to a 1W roluDU of 100 wilb ~edu,nl in a123ML LDMbetas. 9s .4 w0 .1 ~aL ~lHladmaelntian otAlOA 4 prepood ly lais"10 mL oSto 1.0 p&Laotatioa be sAzWvolpme of f 00 wftL mauonol u, a 125 vY, LIVE bDWL 9.1 .7 A 0.01 10 mL py5ai. of to lctlikadaa 0.1 yoAyi . aolaDau oflPDA is Malian to a Ged "plrcapmaemdobfy bringing 100 wA eWMel in a 125 mL LDPBboula . 9.1 .6 Tbe sue* +at ISatiGelttion ealLniame ate rn Lo awmd in a cafrtVnwr fl ~ppmdaoataXv 'e aod an apbA Ox s mlolfsama period of # monlhe ltam tha d+p of popralion 9lZ ilinOndCdilxoboa8oloticllc 42.1 LC11d8re1,$mhvauealordsdaoxepnpatedromatMnotvfadituuon *two.] py'mLlbe4lkFHoawlnlion 9 .31 711e AIIoYPiej is o tyyled aota*c addlliooA eaaaaohalionc may he 9roI ~s aaad~ CMD=bmboa p[Fndllo111rn Qplueaa.(edS1 )DD sob 100 10 b Winds (IMF W s z 10 10 Modb (ML) 100 InD 100 100 100 Find CaowotrWon foahaL~ Lo.9 s .o 2 .0 1 .9 0-f . 1- -v. 100 0.] _- Bqc .af7 t` Page 33 of63 Exygen Research Page 74 of 110 00009"7 Interim Report #23 -- Analysis of Sediment Samples Exygen Study No. : P0000760 Exygea lhotoool Number. P0000760 Mw,rx~w.. vtqo ~ Tp ANALYTICAL METHOD AieAlad of Amdyeir IN tbe pWnpiwqouof Hrtlumooonooie Acid (PFDA) in Sedimeni }by LC1MSlMB 9.2 .3 61t1ro ell aoli2edoa laiEudtId i23-rnL LWE nsmnu-Moldh WOOS . .'t TCA 6'C. up lo six mopthn" 9.14 ApwnwwlUmumW eonowpaliaert of tppdade msybe proprad u NOOK 30.0 HnoYdetUD 10,1 Foci baOSA of eimpkr auaelad (typ1o11y 20 or IM) Mull include an kut au erraamt eW10e1 rd too nrNnted moho4 Wltod at known awemauw (labwonal aplb) se vmilypoocadunl saeovay ft ft eweh. 10.2 Ropimmer ftaold ma le6aa.rs.y duylie.ee. mud xp0Oe. will bo opeocr ed ia IheA~b eruueoe pier 5ov thit ptajea. 11 .0 ae~,owlllanow , 1 1 .1 Wqkh $ j of ample ina $0 MS. golyproprlme oeouifu4e tuba (tortih as an" nylewlia cod mIR wu] 11 .2 AOi 3s II11. of 1Si uellc add, wP, vmex end drloe on a nirt udan ihaher Il .s Ar,amlenae. e.aahy.fl .mu.m-joaoqma0aomimft .. )I A Coodition do CN SINE artridp (1 R 6 ml,) by pauina 10 mL telhLnl 1b11tnwdbyZDml.onpLGpYOf("2dmplwo). DonDtICtcolYmRfnodry 11 .3 Looi (it0two 1W erylt Go dN ealldhioMO Csi SPB euuidp. Diacud /+~ - Iiusle. 11 .6 . Add 29 mL of niqtwool to dre a<dimonm ]e! ia tbe boltom of dw 30 mL e4nOrillye dbe. Cap, roAU end ihlke on e vend aetica shaku fm -30 mimw. 11 .7 CemtriTapSafe6aeR Ja00 qxe, fm -iD sedu4ss. 11 .1 ~tc~nt WemeMxpol m0o Ow aa~ 6PkapbidOe. Cbpeat Cw elute. ' 11 .4 Wpb Ibe oateoln with t ML of melhn". Collect On olaw ad add it m dw piWeaoLaoledi~aoy Il .i. I1 .10 Ceodklcet a 1appld Cle VB ula0idys ( I L 6 ML) 6y tnlssinl 10 ML methanol f011o+radDy 20 MS. of I1pLC wsltr (- 2 dopJa). Ao rwt iet column rw dry 11 .11 Add On moCmlol to- -Z00 inL of WOW =4 load 06 the second canatoned arls~alerw~o. 11.19 BIoY YPil6-4 mL IDOl; mlAstqol . Collect S ML of Alum* inq Fadutled IS mLPalype:pylmo cmuifhpsuEae (Anal volume . SmL). 11.34 Aaalyse 7ropleo udn5 eloxon"q LCMBIM4 . pap 5 of? r' Page 34 of 6s Exygen Research Page 75 of 110 00001,98 Interim Report #Z3 - Analysis of Sediment Samples Exygen Study No .: Pt1000750 EXygen Ptotocol Number: P0000960 mVJ . trrsw +IwelNMMInrvo00171r +w~'+W ~TI~OD MWwd odAaalyrfef~rlhM Dosadwtiaa ofPw~wamoootaoic Aafd (PFOA) in SedimwK by . LCJ11SM8 12.0 Mammoyraply, 121 WW 0R irWnr aooold of eerA woodri, aeaiplo and 1bdlnd etrnpb i n to the LQrq!M qsl~wot. A eallbrriioa oatnderd must pieeade and follow alt apdpod oaeplw, 12.2 SuodadaofPPOA ooeoqoodby a uiee~ Aveoe~wre oonoentrocion k.rJs owrtbr mclodd muao~Fy4cal aR . 12.7 Am ealin wt of enpaRrd twflkstioo pandarde most be inoiudod st tde beginning and et Go end at o moplo Iet. ShndYds owot be imvprerred bwuoW *rcy 5-101ppploa AM ri Wminflvo, a mtiea wt of calibration tlaadardt my be a4eoled at do beptninE of a sot folbwad by calibradon Awoqrda ineolnpeeood wwy Sdtl erWls (to woman !6r a snood s*i or ~ In *ON am odiErflfoti aYodolda soil b0 1116 rag ead last i%WbHe m t mosp10aA. I2.1 Ups Haw smdoa awes Or 9rtcda7ioo . IJaer wndud eurns ore ' Oeoreald" br to ainp" by Flow nFathm oft Ion wp3Mn= ofpeWc am vaeita ' oij, abaidad aMrwtOtotion udo` MarLynx 3 .3 (or aquiraknt) roilwas a30ak 12 .5 3napls swpaaee aLoold ox nosed, rmodtud raapamow. Any asnDkr thu roroond o4edrod rapaoaea shoakl he Raduer dilpUd and epmNyoed . 13.0 Amepwm t7dlwii 13.1 CIuItObAtVM eaWl IeoW apML of adeusplt ion at 369 AM Mao rpMrs 1~ ofA3 IIOqI. 'ILe "11 am PRAM Catiopoodi to OW PFQA saion, whilc the daoliktp' imt (3d4 aow) npreeoop the Ion of oerbon diuaide. 132 Mathod blplo low am eoptaie MA 1[ Joveb Veasor, won the LAQ. if n Me* oentains PFOA rt ler+lb peNr thtaa 0~7 aymL, then a new Weak aoesPL MM be obblAed rnd go mtiro ret must be reixvaoud. 13.3 Aoooreriw of oooaol *km md waaix rpiipo motc bo bnweea 70-1)0% of dtdr blrws valom, iY a ooonol apW A2b atYide the aoe,pta41, 6mitx tAe utio got af srmptes alnuid be ioaxvec7od. Any maaiY apit. ootsid. 70. 130% ehoni! be o+aAtMaf by 11w tauiya to detormEoe if tbextractwn ie wetaatwl 13 .4 Any odihratieo slardord bimd to De a rtufrtcol aW1Ur by todea tde Nuj,c aAr Ta.. my b " w;We+ bon, de oaioulowom of ft eweradon curve. Howww. du OW oumeee Of aaoaa wlbneioa stoodartio thin could be .alodae men not axeand 7on of thr now Mmihar or sxtradea .uoeaM, 4004 ' 13 .6 'br amoledon ooollieiwo (x) ibr vallbqtioat oorwt pimeued muwt be ZA.992 00 20 .%S~ If aaacsoan raulb A11 eutttdo thae tisnils. G. oppropefas Rptt Move be Irhm m aOwt ineoomwn o. peration sod The nuosindi aOw eelsMeM go otfo"lew abouId be remndyaad. 1Nriel7 Page 33 of OS Exygen Research Page 76 of 110 Interim Report #23 - Analysis of Sedimertt Samples Exygen Study No. : P0000760 ell Exygee Promcol N: umber p000p76o HqynYAwrel . . . 1406" iMbw VODYIld2 _ - 91HALYITCAL IiSLT80D AtellodotAslyde fix Ow DWrmiwlim ofTxllroxooetimoic Acid (lAOA) in Sednan by LCJA48W 17i iweEm firr Ilpw-en wodxidr and xwmptn =t so dril! am liun t416v"q iwxlydw! nn ifnlel4nn umn driR aclpy Ihds lbnit wilhin narlytial r~oIbwdw wt ow~D~ raalyad. 14-0 Cxtoubdeoc 14 .1 . . use On 44u cqpwrioa rora1c911ne in uaouqt pfPFOA 6mxld (in r*mL. bdnxd 40 Pest uwy trlq 60 rlmdud cWw (lipnr fqrordaa pkxwnen) . bmaue6by0r Mw Lypx SnOwoa yroptw: PPOA Fpmr( fnofmL) -OAWLAML-iafMAWl . x DP Stops m' ~ 6tlofbywlieh ebr 1W1 vWumx rvu ~tiWa4 if ncanry. 142 Pa wwplowfERi6d wI011no~w xouwompx of PPOA prior lo axbaction. uK W 16suxanjwlrWee le qbWw Ow ywewu neowry. kowY(%J' (som .n.ir~fbma(~y~[.1- mxlyl .rwna .Aar~""aa.e c~) ;a~xw1(na~.)J .loo 14J IJis tbs lallnwio~ quplmi lo copvrn $w want of PFDA found in nbM~[. lo aft (tPbl PPL1A&vnd (*) r [tjOAApP4(flpW x Em) - .val-e (s all 37 . UM* wai& (5 b) 1i.t Uw Ibs E~o~rm= xquxUon (if aeowry) ro catluuLN tlw atnouut qf PPOA 2wn0 in ppb 4wnd andry wsitYe. pFOA /bund (PPb) dqr+rai&-BPQA fwnd (Ppb) x ( 10016 f bW wlidpKll rhp T.r T / Page 36 of 63 Exygen Research Page 77 of 110 000100 Interim Report #23 - Analysis of Sediment Samples Exygen Study No. : P0000760 .00~ A-~ Exyg= FrMC01 NUrabgT: P0000760 ANALY77CAL METHOD "adodNwaor VMIM M66SA OfANOP66F OnA Chm byWJbWM Uld (TFOA) 14 NO AsWvWdTmi1nxfsdW. Amm Racard, 306IRSwftdkDrivo Sim C*U^ PA t6vol Appm"d ft raw COAMIly Twb"Ladw. azFkh-y Ito. P-Ma", op-dom 1hyso ONO,, Exygen Research TYWPMU: I Page Vvf65 Page 78 of 110 Interim Report #23 - Analysis of Sediment Samples p. 79 Exygen Study No.: P0000760 /^ I~` bxyBan Protooai Nunber. P0000160 la"hedlh..r.. vooovu Aw+( .rrrncAR.NOMaa uphod o(Aablis ks pnpnennziArtian p/'Pee~paraoctamic Acid (PFDA) ia Fbb and Clamsby LCJat31AS9 1.0 I)00~~ '1'!tio m~od In In w syloyed Lortlw iwlalion od quntilntim dpmComooctowic WA by Hlyl Ietrsitooe Iiquid CkomuoF4pigS cnpkd lo t tandam Mug 9p~nuov~ollicDdsbrll.dAi&wJ in Gdi rad drm. 20 3oRty 2.1 Alw~sobr+" nfrlafiodarlrpr~ti~. 2.2 Cannhlhr opAoAAb MW Who bwdleS any donical rurpapm uroly 3.0 9asipb Rhubomrm 3 .1 At Wed 2D s oFlwl tmpk be tfihfelia. 3.7 - "11t Ib0Y10 ho pro0nundl6afnre abnc8an Plane Iho fin= soenyk in n food pi;ooeros m0 homoooqiu w11W dry ioa. Mw tM staiploa La oonnmns Iqd were apm In itx.at paap ovwnijit 1o n11ov fnr cwbon diaxidc iubllmftlai, Sea pul place Ow wrpla la fm- reprw wuil um or t1101)"10, 7.3 lampb collection proeoduns -ill be specified in the sampling plan fcr this FMj- s.0 t,CqoWs wed $undcb 4.1 WOW-F1pLCp+do 4.2 AoeWalvilr-IffLCyade 4.3 Crbon(120400mwA)-ReWetpWe 4w MaOml-HPLCVa1e 4.5 eiGa P1(&200 maL)-ReoOM osde 1.6 F1od3il(60-IOOnala-R3ysfmtptadst 4.7 Supefatan LC-Mh-xeYew Fv4 4.2 1-0cfimol-HPLCy3& 49 410 Inr6Avr&o0x&hwiocfnfecwg-ilA.ta"-"ae&.I&w g.ae 4.11 7olvrnn-lleqant Fr& 4 .13 AmmaolmnAaolm-A.C.E.1wqwrcOnde < .13 PaAaoeooewwioAmid -Si;mt-A]ddeh 5.0 Lroroonsr aod 8qniyannl 3 .1 A Will; pedbruttna liqwl vWwww5ropit aopabln of pumping up w 2 .e]v.ru .quirp.a wilk o,nai.blb vopn%o iqia,o, c,pWlO of ilk)snEnB 3_200 w[. aa~wecea a a pad3en>d3r ~elromitatiLGiNSfMSI. J~2 A deria to oollptrsw didafaryak iotbsidaa and Yuacmlman . 5 .3 ArWy4aa1bd3perCmpablosteendiojb0 .000D1 p p,p ; ra Page 38 ojdS Exygen Researeh ( ~ ~~ j , Page 79 of 110 000102 Interim Report #23 -Analysis of Sediment Samples Exygen Study No. : P0000760 ExyganPfowcpl Number. F000076p Orr" AMMA 3i~oiof u0aa6rwobKvoumJ AxwLYT1C4LKCTAW !1~ Mw DMlaad~rlioe of /sMo~Oaoi~ook Aad (PFOA) tin PiYt rd , Clm bt LCI1d9fAid !A 11Nrqsvqwrmps. !d 711n~i :1r. ib 125pLprrwhrpdarls. 5,7 SO mL Liqpw*kPobAOpy]Womodfhpluba, 5.3 5.9 5.10 15 al . iiPwhlipolypmpyla anai6oie fnbss, Dbyaatl10 wiepn,iMk(SO " IOOyi.,1004.00uL). .175~pSl. LW81lrcM.pl0olp peRlY. 5.11 2 mLOIwrMlWr7d W $12 Diyaa4lbpdy~aw. 5,13 MWAPOOM (toaioooILL .e63a1ooJ&LaMmailporr,bcP. . s.w 3,15 swAseef+iae. .u(n2a0rMWO.(:SUPWM UL ma W057177) . 5.18 cOe0ifYlLecWOsot.parmhys0n1.polJWapylen.wbaa 20M W. aa cwMdav*kcsWAb . a.t Ae.bUwlcoWwptneyhn, RP (xerimae sc4matk). il MM,c so mm. syi (rn~: 825654aul30 42 T*qMwM 30'C GS FfoFikl~6~s fA) : 2 sM Amonwidma Acaerb is Wuer 6.6 Moti~i~IB): )Ala6arol 6.3 GwiSmtTroywm: T1pIP.L01G1 OA 1 .0 IA 20.0 22 .i MLA 68 65 25 23 LS Flow Rabc SSB (m11= 35 0 .3 33 0.3 75 0.3 75 0 .) 35 0.3 L.L lqjeplasYohew : 1S yL ~awDoma~wd ta r muoh u !0 pL) . i.3 Qwpafatioe~ PuY Afe~-eactwnd ~twlad crlilus~ieo curve. Cf Rat Tinx -23 suoutes . 1b* "" WpMou6 an blooded o bauide md may bt etunaed m ardor to oyd~mme P8[ .C ~ynsn. hp ).t~ Page 39 qfO Exygen Research Page goof 110 000103 Interim Report #23 - Analysis of Sediment Samples Fxygen Study No.: P0000760 /- Exygen Prooocul Nmqber. POU00760 hrw~ MeOwd Aaalyaia M.uoal+=1e..vvolm ANA41c[!CAl. Jd>cl80D 4aDalrftdoel{oa alPeOopewxqopie Acld (1Fw In Pi~h rul Cbms by LC40lA44 7 .0 MSIA6flyua 7.1 Ltode SInYOe~y1JKaYw t~!)~( moda, oinai4eio~ ~ l l -~ 769 nh fu PFC1A . 4he abow ea~lhwr aa inh~dad u a ~ai4a md ouy M elsoqd ia adac w ' ~Iimis1he11SAOal+twa to tnpnWmofSoAmwu . 9.1 rw611 .Pen. Rl .l ~ xou aumcoiyq, .nwe i. ...,Q y wqweo br ~^e a=u of ~ aaw to Sooamt ad.ratx. as - llre~woa swwia~ 0.1 1 2% ooable add to mNhaMl ia prepaced Lr dijnivasj 2 gof aeorbio tcii iar 100 taL ofmepuuwl. 9.7.2 3Qli D~yldiebloroillbu Le bluan is preprsd by Mip~n` 3 mt ., of4imeftldiehbta3ppa too llnd volume of 19 mLwith loluau . A]wrwa valumer auy hepnpred. 9.0 Atradrd Pntpnaiko 90 PAmfaNSmrJc'Potef9c+donsolwimt 9.1.1 Pnp=o a stock phtioo of-100 yySoL ofPFDA by vsigl<ln; 10 mg ofinalylial qaodud (tuarxiod fnt pwity) and diluto co IOD mL wk1A nrlhuel is a 125-aa1IDP8 be" 9.1.2 A l.9 pyhoi faelidczdon aohrooe oPPFOA is pte,pirad by brindina 1 mL of fLa IOP+yhaL aalrtion bs Erol roiux of 100 wilh nxaueol in s 123 xnL IAPEbeltSe . 9.1.7 A D. f pOImLfaudoloionaeludofPFOA iaprapvM Ay Mating 10 qoL oftite 1 .0 pyqL oh" Ip a foal volume 9f 100 with wamol 4i a 1Z; mL LDPkbeqla 9.1A A a01 )tyML 10tUMalloe aolulfoa of PPOA is pcewrOd by OrinPfng ID mL o! ft al IV+d. aoLkioe to S Find ypluwpe of 100 with umad In t 1].S mf. LM boldo. 9.11 Mw iOOtk .nd Jlnilicadoa whwoma an to be .mna ln arsSSZm.wr .t aPylo7tlAtEdy "C id w alahla fm a nnntimuW Paind of 6 ~ths 6a2 9e d,tsof PnPvaflm POW 49r, /" Page 40 qf63 Exygen Research Page $1 of 110 000104 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Exygqn Protneol Num6ar. P0000760 s~a+wwer 1a.emr+rmw woonu AlIALYYICAL IAIGYSIO ) MAWnfAnapfalatnhsDlltaioYior Of?" uotooetmue Aoid(pFOAia?Ycdulid CINU ayLCIMSAt! 9.1 SWWadC@libndon9o)~dma 9.2.1 932 31&989O qlihrntio4aftedan01 wpapered in methanol viadilution K11e lA )IDImLlbetilleulan aolutiml. 'Plea fo)Ivwityis appioa) axnnyla additlmwl ooocswaoons me;, be ~klwWatl. Cato0010011010 otpalifealim YuAnm nilnudb Meet! Gaaotwatlam idtCea~.(utiLS .. m- ..r . fm[.? IuMLS 1:0 5.0 IOD 1.0 _ 2.5 100 0.03 Q02$ 1.0 1.0 100 US 'o lQQ '0.024 i9 100 0A1 GAGS DAPZS 0.1 t0 100 0001 O .OD! 10 100 0.9005 0.7.3 SSOM W oa5Entian atsn0aade ia S7S~mL SDPSi aanow~quth bunks It 21C to 451C. op to Moaft, 9.2.A klIiraYe volunuf ao*1d1ooaocn6ttioqs o7 rtmduda may be prspned as 40011101 . 1.0D BatdtSalUP IG.1 Bad Ukh at wwppSw nmactad (typkally 20 or IM) Mast inc lade u Iron OW tmUadd am1Do1 MW two ucrttlaled ccceola Sb[d8me at lo~ nn11C011VYSd1/ (~ OOIIRW JpIYC~ to YIIITI' A~Il t0n0vCry bribe UlICII. 1Q3 Rpu$~4 Sor Ddd dd liptYOry dtyllGtp altd spitu will be spedfied ia Iha qoaiily aasoraaoe 00 ]hr We Proem 11 .11, Slmapls Ilibaotica Sl .l w,4ah 3 s or twne mmpas InoP 70 ad, w1YFwylane cwnriinye tubes (kttiil' w aae" iaplaoc lid wd six well~ III Add 70 mt of aeliollisilr at 11Wb os a wrist Woo ahnkw flu -13 minuwn . )1.3 171or+toa tvibaa in 0 4wsrfar-11wur. !1A Prek well aoalltitiaallhM Ev8 mba ad lUWm IM pranlbsDnd nstkx t1.3 h01< the 20 dLL 99B lubr m nyursa niJh 2 s floriul, 2 j silica pl. 2 ` *46m, ad 1 ` I+GMi. CendiBen ft aolumM vilh 20 mL of n1etbrnol . do 70 ml. of wtanitrSlr. Massed al1 washes . Do aot slow do eolamn eo ]IA sky. 8i]mits On 125 mL pa4aEWpad Seeks by lledsy wilh IF6n 30% d ;nmQyIWehIMadlrk io Bolueaa aolnSloou Rim tba Auk -kb lolaeaw G. us AIIOh'W by iMIh1110i SI11fN 1]R7M)- Dh' 1/N We COplylOta)y Wore uat, :Ihf 1y IINAFAIaRidl l 10lIYI YflaWyln . f+ye S afn Page Il of 61 Exygen Research Page 829f 110 Interim Report #23 - Analysis of Sediment Samples p. 83 Exygen Study No. : P0000760 ^ r ltcygea Prnlocol Number P0000'760 6t1>Ir1olYkb ~ ' IdrlbdllambrYpn0171) AXALYTICAL hIi'li{OD ) Mehod ofAs" fm do Deoecriaaion ofP Chma by LCJAtSIM3 ' Acid (PpOA) in 1Rth and i l .7 112 119 11 .10 ! ,1:.-I12 11 .13 11 .1 " ]i .1S 1 l.li 11 .17 1 t .li Cabifiaa San 70 mi. pofypapylaw tubw ooetoiuinO annplo w -2DD0 ipm tot-SOmiowtea. DIOYdOw tdnotft IQ aoolNltlaaad S!B aotartal fitEad iqtide an mouth of Ow poor-tbtgd 11qK Collat the eluft in pte 123 mL tilanima perr-tnape Flalk Add 10 sL of aoalonltrile Ia Ihe tan7plo in t11o $O mL carttifup tubc. HoMqwi"sathe Noon No pWuYoung a IbaNriirlbr -30 aoeeada ~xW rinac q tbwnoadrrhh -10 mL ot rptooitrik in4 Ute ntha. Bldlr Ow Mwla apirlar-10 murtrno awqiq-actian ilmka. Dlua'9n tubw bt oGeacr~r- 1 how no". CHN111bW,a tho SO 7at.polMopykne Value ecnpne"raepie at -3000 rpm Mr-IOmleolet. Damn! No wtraCl abla dip PAW Spa c. olumn Cotket be oluote into Inc eme yep.aLapdd tlnk will caeabine with the slum aom the initial IRKKfio " Mr ZO rsL af turoldidle 4rouy+the bPE co7uma ud cambine the tlwx in tbe ayoo ~u.~l~rd fla+k. Add 3-4 dropt of 1-ootYOl In, the exVao[ In ft par~ahapN ppk Bad erAonu undwodyadussousy arawlr dvaporalmta9 <"0"C1. tdmlu Me find voiuulq, by addpQ 2 m4 of 2% anootbio acid in teetbaod w ~htp~'thaPadAuk Ilnd fwift bdlWldkqhr4. 71aoaGrlhaaccwaqa0>F1pLCVldIaednjdlrDaaaWpptpatt . Anatrw eupkd apial doonolpptsy 1.CJA481Df8, 12.0 chfov+laywhr 1Y .1 IqJaR "wniammm afrrL alaadad, umpla arl Ibridifiad ample into the LCJALV111S rydtaa. A cdlbudaa a4adard mut peaoada nad Ibllow sit aoalyaad -=ON. 12.2 1fn9mi4 edFPDA oarn4andLtS o at leat live or nrore ooacsatratian krelt curbeinoloded inasaad)ruaa! aw. 14.3 An adwaa of ntihertiadmdeia mum be ia]uldd a ebebesinwns;aad ol dlsandnt asw~pla aat. &aodadda nwat ba ineenporoW batww, rwry i-In uiupka Ad to alktaadve, an w6rc act of calibration aandnda may be ipdetad at the beginning of a am daDotrld by alihntion Wndenh IlnrrparoN sverqS-lo rraplu ge aaenmt 1braaocapd ex oi rtmdfrdiy. 1n aiWr wit, ealtaatiea alandarda swill be DIP Not wA lap iqfaetion in a aamp4te . 12A Un liwc pnldsd pars 16or quonlWtiaa . Licau oftedard wrvr vc gumlod forthawlyleDy Ilmp royerion wk4 11% waiahactitof paak am randd oh'ka6ein +medard oomoanbYian uoqn ; btapLym $.3 (orequivalent) ' aOteWQaayalYa, ry,sota Pap 41 of 63 Exygen Research `ft Page 83 of 110 000106 Interim Report #E23 - Analysis of Sediment Samples Exygen Study No . : P0000760 B+lygao proiocot Number POW760 syrsa+~wea r~.r.eH~vooalru ASiALYMCAL MS T11Go NWwd ofMdysis 0lf the Dasedebdan of )aAnqooqabofc Mld~FOA- ) in Fish &W Cpme byLCIAl81M9 12.$ SE*! IllpnY OINIld II01 Oio7Cd IIfY1dYd fllp<n11Y AAy soBlpla IMt eaori standard responses should be 6u4rr diluted and resulyzed 13.0 Acaptoieefkllatr 1] .l CkooNeo~+m mult Jlhow apwto{n dw/bw ian u 369 atmu trom a parsni of 417 was. TL 413 uoupacenl Coqo4opdr b 16e PFOA anion, While Jhff dwrtriaa(369m$rbPr- ~Or lopof eblbaa dian0e . 13.2 IiGOqd Woft most aot omu{e lPOA p Iwe1s Flow IbNS dbs LOQ. its, bimtkoomtdbr PFOA a1 taseis FuWtiup D.6 ppD, thm n aaw blann aample an* toobl4iwd sold ow Oaks so Mug be V&4xbmtod. 1] .3 AowYwiul of onWtai splloa rd mw spitm must be brtween 70.130K of sMfrlrsolrb rwAlr. Ss aomool apille Wk ouNde the nooeDpiWe limits, the wihe wt ofmwlw ilwldlrnsoctrmsnd. 13A AnD'salibmiow aaodud lbaad oo Ls a Mvdfical owliK br win ft iinla 1'aoe 1wr, mV ho audadsd Own So eakulYaun of dx Ca111Dcuion eYrrs. Uowrra, Ilu 1otY number of edWmatiou standards that oould be excluded must 10 allowed 7D'KatfLe OW aumbtroiaundards bysolud. 39,5 S3s eondatiao snldleisS l3q for adi6nlios "via Illustrated, mm1 he t0.990 (0 k0.913} ]I COMM raWW fdl outside tbese limit.. dl, applapfilfe 4bpi most bt taken 1o adjust iuuhumeet operation, and t1te rudnedsat We nlwbbt bN of wpia rhOYW be terlalyted. 73.6 ielOtim dnw leHlwl standards tnd nYl4llq must Aot doft men [hall # 4 % wimiw rs mdyuW nm. If reeeeulon limb d74fl exceeds WO UrAit with ill ap rlpdy4od nut Ik1ep do wt must be nurlysad. 14.0 G"skrolYiau 14 .f Iktdie Poi16xl~qwilwl9o wkulbta Qleallowt of PPOA fuumd (in nymL, based on ppk K0111 wlnj the abmdnd ~(liaaar reReniuoi yuamdasY ;IalMedb7Mrofd1M WasIIbRwfnpnptas: PFDA bad pIpImL) - iPM men, - lm6m=l do" 14.Z Vsa tho fcllolIFiRS pWRipa 10 ooAwu Br uboum of "OA found In nVnL to 11041114 PFOA loved Wpb) " IPFt1;-feue!le I_~,~ ,1Tp~;,~1 v,Leti (ML) : DFl -wig wwgw (J) ppwfaotasbp,vhie! dw tlal volwtw wu d11utud, it nrowory. rqe r eer tow Page 43 of 65 Exygen Research Page 84 of 110 a' Interim Report #23 - Analysis of Sediment Samples Exygen Study No . : P000076fl 8xygen Pmlocol Nwnber: P90007b0 9" x"tt.,rwor .f+1WrM er*"a rrb.,vooeIn3 . . . .~x~u,mc.v.ruruoa teeaaramcl;m.."daw,o. it.rcoMlslr~uoactao~c ncW (MA) m xh +na t4.3 Par Mqop1K fmefi0 with Imo" Omnunp of PPOA pdar to extroaion, au 16=fo0owmj vquriou ro oUculuo t]x PacontTwovery . tteoowry.{%)+ ~. EOUlra~yEeICuod{xesyMiel~-ddw~ldy(b+f~o1m?iiuoootrel(o~1~ ~100 Exygen Research Pqr I .ti Page 44 of63 Page 85 of 110 OMIOS Interim Report #23 - Analysis of Sediment Samples Exygen Study No . : P0000760 Bxygen Protocol NunaEar : P0900960 ANAtY1ICA1. M6THOD Itldbud M": Vppp1784 Mum *tAaNyNi hr MDrorslziqoa s( Pwflsionmfaoak Ado (PTQA) In . VKebNpq by LClMB1M8 Aaab'6d TstinY F.Wiq"" BxYys R"wma 3058 rt,roa Dr1rs sr.ee cAnqe, PA DUN AM-a By Pw1 Ctqwolly TWuda1 Lwqr. LC-Mf.BxyM RraKB s6hLla~ DW A?~ Flahrry Dow Vio. Awldmt ~~por, ~cY~w A~ue6 ToW Po`a: 7 /`` Exygen Research Page 86 of 110 0001.09 hiterim Report #23 - Analysis of Sediment Samples Exygen Study No. : P0000760 Exygm PrOtooot Nlnber. P000076o ANALYTiCA1,JO HOD Fdatiod of Awtyds 16r Ihe Dawim~uf .r~fr:p~oootanoie Acid (FPOASi~ v.dniiiao by LCIMSIMd 1 .0 $tapt T.V memod u a. be .mptahw rOr ap boltUm oil quanticusien orpavllunrcocunoir aoid by Hith PnfEnaaxe Liq~dd ikro"myW6y oouplad to a rK+dam Men 9rwtraoWia DKoolor(LGN31M7) in vgomim. 2.0 sa6ary 11 Alwkyobaerenlbiltarrmryyraeucea 2 .2 C9RN! 1he appopwb MM 14lboa lundliae any cheroiul fix penpq aarenJ pnamoipqt. 3-G EsapL ltaqwamwal 7 .7 At kW 20 S ailrt arotplolos eaprllmo. 33 gmsplea dlwld qe.proowad Doors nlraefieo . ft. tbe Enasn ~pnplN in a 11443 aoopl~ wr1 bdmapPdm wilh dry ice, plsoe lly pmpla m eoMtiam nd laW tpoa fm Evrm irofW 9+M& io dlow !or ouban dioaide Obi-Gm saw now plars Or aanylaa in 6oua abnya undl t'vae or " 33 Stpqsls tollo0ou pmoldlum mill bo ap,ei8ad in IhM aamylmd plan for An YaRIwL 4.0 3tsMto4sdBaodarda i.l Wnx-HPI.CVade 9 .2 Aoatad4iJa-NPLCUnde 4.3 Clcbaw (la0JD0 mwsb)--Aayrl yrads 4.4 Ma111i17D1-HlLC ~~rdo 4.s ae srlde u.;.rjl(ds(0s-a1X0oommr.er)h)--aIaytrqaaerygaid..d. 4.7 aupcicl.im l.crrx,-aaapnvpaae 4s i-oednet-1u7.C~.d. l9 LAaeaAie aaid-Rr~1o[ pada ".1D DLuatbyllGblaoai(w-ftaadaxy~b 4.11 Tolaeae" Roadeneard. +.1 : Arainodum A . .w- e A .na. ae,llan c3raao 4.33 welLwmo .d..ianadd-sypo.v+ldrfa4 5.0 twcmaaR md eqalpromc 3~1 A " pellBrnAplot liquid tSrpmatopapb capable or pumping up io 3 "o(raea oqYinld with a widte volwno (njeolar cnipablo or iraierdny, s.20n PLeonrectwllo aOadrm Mw 3yook~x{LCIM91M8) !.2 A dsvioa to eolfeGnw ns rorpaak iouWauaa upd qxantlupw 3 .3 Apnlylied balauee oaabp ofwadind so 0.00001 e. Pip aar+ eft Page (b qf6J Exygen Research ,it Page 87 of 110 00011.0 Interim Report #23 - Analysis of Sediment Samples Exygen Study No. : P00007b0 Exyg= rvooxotxummr;c : roooo-r6o r grApoacroor Hae.t21vq*.rvopor,K Meuw6 DrAnYHi. her to. Determination orrxeuramaoneole Acid tMA) in Veipeiutan by LCxs,W sA 5.5 R13O5 aWnt.PraapwacaOk+r4.w finks. dm Saml.mrpoue(.p.ypcopyLO.coma+fay .aae. aa 15mr.auyeueup.Wop)aee.cena;figChb,r. Si DSAOru4lrmkmpoer(50.t0001.10040GaL). 3.9 1. 25-m1 LAJBauro+y-maa6 bvplrw. 7.]0 1 mLcIUa FIp1.C viajlot . s:u . ~~(l~oo~iooayt.~alalo6vt~,~lr~qe..aawy .- s.ls 5.14 srewc.(mPr.?.tsrpwrac .l .ra.NOrrlr7~ wdsMOOD .],.u.. 515S Caemirtps etppbleo[qiamft 30 mLp0lrpapYlare Mb" E12000 epm . 6.0 Chamuop%Phio d1MM 6.t Analytical Co1Wm1o IhYqlup RP (Reyltooe 9ciuft ] .1 avn x 70 a.m Sp Q'A4 :12903-OS2170) 4.2 Tpsat,rrt ]0'e b.D Hari6Phue (A) : 2 ta1H MuooaWm Aceue in Water 6,a Mehilc Phone M : MV&Md 6.3 Wrrdirot7+ojom: Timtlmw 0.0 1.0 1.0 20.D 23.5 L' A 6S 63 25 23 65 Flow 34" 'hH ~h~o! ]S 0.] 35 0.3 73 0.3 75 0.3 35 0,) 6.i lqjostlogVaL=r 15 ILL (or be i0avuat to ea mueT u !0 w4 6.9 QOtetibticec Pat Arm-~fjsdsdcall6raHoacurve. 6.9 RanYSnwi -23aolawca. Tlw am oenaidony are idcaded as a Ode and may be ohinsyed in order eo opdmite dr HPCC syrpm 7.0 MOM 61I1am 1.1 Lfa& Bla4uWey Nepliw MM modr, ruoakoriny 413 ~ 364mh far FM nr )or " Page 47 of 6S Interim Report #23 - Analysis of Sediment Santples p. 89 F-zygen Study No . : P0000760 ExyBna Protocol Number: P000076p >~'"~ " ' -- ~ehnoeiveabr.vDODi~~ I - --_ ' . . X1vaLY1iCAL ~1t1'!{OD bdwhodafAaat,raii 16r Ih~ Oe~idn ot - by LCIM3fMS ' Asid {PFOA3 in Yegcunon The doto0n8ilism oa Int.dal a a oipe and terybs tlnnpA in order io vpe~i~tfto M~Ad eum . HA Fnjentioe&or$eMfios 8.1 Mobila Phaee 1.1 .1 . 2 146! anmaq~lm aoehp In wpu he peapand by addinR D.04 ` of ~oei~ostwlaeaq 1000umLorwWe. 12 sxatAraan sawhaae i.Z.l 2% aKaEio .dd inanefLbot ia pnpenli bY diWOhbu z g cr.tcbic radla IODeoLoraletlaqol. a .i2 30% P1mW0didlloradhle in l0iw*s it Ra9ilv+d by beiadiej 3 mL oddimedlykUJaallano m a $M1 volaula or 10 ML with laiuwe. Ahcnalo,olomamaybs prepaoea. 9 .0 sma.d Paepantion . 9 .1 8hodwed SIecbFeetiBcatiow Soludoa . _ - 9.1 .1 PRpan a Oak aoiKlon or-100 yvwL ofPFQA by washing 10 ng Of nalY" OEWNd (aORMlod tX pqatY) and diHr1c 10 100 m I, with faelbraol in a 123-mil. [APE baede. 9.1 .2 A1.10 pjlynl . Ioniticaflwsolntbn of PFDA b1ve"red by brinyin= I i7Q.of do 100p. >%1 ealoelool in a Anal va111moor 1G0 with nectleareol ia a12SmLL[X8 baqe. 9.1 .3 A 0.! py'uLAvlifidtiaaoolulios of "DAis pmptnd by bd~10 ' eeLbribe, 1 .0 p/JmLaoioGon to af" voloma or l00with bedhmo I in a lss m1. LPl?8 botqa. 91.1,4 A1101 yatsL 1brdAadaet wluttion of PYOA is prop" by brinpn ; 10 ML of flW 0.1 Pt'mL eoluEOet lo a 3na1 v6luwno of 100 with mrbaaol io a iX7 mLL171;E boWo. . 9.13 Mine mink and kitidcodaa aoludow am to bee stored in a rikiprseor n applaotimately 4"C and as enable for a maimmn period of 6 montin Am to dotofpreparatioin. 9.2 bmdud Ct1lEeadoa Eoludow 9.2.1 6C1Atylir9 eaL'Maiipn ahmdpde are ptapve0 in M94heinal via dilution Df f1a 1.0 L91mL lbtdncviml folotf6m peyaua " Pape 48 of6.1 Exygen Research Page $9 of i 10 00011.2 interim Report #23 - Analysis of Sediment Samples Exygsn Study No . : P0040760 Exygen PmpoQol Nulclber: P0000760 lExypo AwwcA JNWOmd Nunisr VCQOI7H u~~u .rn~u, l~u:rlloa hbmbd ofADqyul Ta Iho PrUQmbtNwt OfPerAQQraoollimie And MA) in Yeptition by 1.C1N8IAt8 922 1U 1D11tnrwt In " typlCt3 outpyrl0. aWiliOIW OpoQ00Q'aipWi tn0y b0 /f~YdafAMdad CauwnkatiW v(Fp0(wr'OO0 VohOar piklW b 7111161 Clresvtrrtiim laiutimlrshai.l 1m1.1 (mLl fudmt7 1 .0 S.0 lop 1 .9 23 110Q IA 1.0 Fp0 oOnaMs IQ w 190 +Go a0.o1a a tloa 110900 9 .2.3 Slen rl1 etl0x.tlan >t~drde ie al R'GO 4't up lo six tawtGu, I zs-rOL ball 0.02f QAI 9-9003 0aQs 4011.0-0O1M t,ppH aaerouamouNt yoeua Q14 A11rqW velrous rldeane~phutiow af wndatds mayb~ pcapvd r aedd. 10.Q eakltBetUp 191 Bath OOQ* e1 Srnplu OIIIne1W hyDieaOy 20 OF leos) mluc include 61 3ust m0 sfalimaw eeard ati two mnnoNO warub fanil)+d sl fswwn "I'MlotlwlQ (" ~01 tqiloe) to vvrily preoidanl reccimy for the buck . '1012 Aoqtli[qormtfOr9eldrWlaboxalarydupGaAmlaid Wiicmwill6efpecified in t1us+IwGty aaaaoa Ping IM this pavjeot. 1 1 .0 snpie s:lnQdos 11.1 Woo S Y of $osm aimgiu iaa SO mL palypaopylafs cenui$t=c tuba (tb1t1[y u iyabd, none Lid and mix well). 113 Add)OrpS.pla"cetaeioiibmWahateva=wrirtaction chaterfot-I7minuta . 113 Caut'iBuK~ be 30 mL polypopylme tuba containing tample u -2000 rpm 11-4 IPN.~-t1Ct~m,ialeonusfofiru. W de sPatuba ad r;l .me.ateyo,w-awp.o A.akii. 11s r.k do 20 m1, ara mw. in mclusnt. wsu s a noaNl . 2 1 *i1w. wl . 2 j erem and I g L:C-, NH CoamSioQ Ihe oQlttttmw with 20 m1 of nuuhuI, 16eo 10 mi of ~c~ooaiCllr. Dircmd all wrhea Do aM allow UK cqlrmn to dry, 11 .1i 9iWdse 1W 125 taL p0avhpOd fLda by litaul= with ft 30% ttinsdtylmehlovOdlmr in toiutw rQluOao. Rim thb nook with Wainit *AM ilfOowed by tanOmttot (lhm tinlal). City The Atrlu coaqbnly bubto uee, dtlvpy dr-deyinjor v1Yt anmm ofnitreywi 11 .7 DOuot ilhisacpad w oc aawratitwrd SPB aalu~ Qaed in" Ow pwaqb at lltr Pwadiaw Ilult. Cblbet t60 alLOer in *e 123 OIL riUn[1"u pw-Fbyx 6WI. Pqc]st' . Pop49 0163 Exygen Research Page 90 of 110 Interim Report #23 - Analysis of Sediment Samples p. 91 Exygen Study No.: P0000760 r~ Pxypea Protocol Nwn6er. POD00760 lb,ygeaseewA . . . uediea Hur.e voout 154 A .'+IALI~TIGA1.31ET80D OQolhad ofAdlye4 to On DdumioeliouofTfeihlawaetauoie Acid (TApA) in VeOeution by LCJMS9dS 11.0 Add 20mLefaoeleoiLVilem the =epb in 9w 50 nd. cui5ye tubw 11.9 'balol6r moeple spin fn< -10minutee on a+wiel-actim d. eter 11.10 Cfah30ye tLe'60 wi. pob0eory4w ]oDa oooWnInS ample Y "2000 epm 11 .12 Read me ~ad coo1he woe APE =ban (bllece the ebuft into IM sow peerAlpe " ant MA oonDiwe wHh 04 elueot dnm the Ini6a1 11 .12 11 .11 31 .14 11 .15 11.16 luprew0s t1 .e Ibmufe 11.u spitz Add K *op of Waned tb Oe sdeart a do par-hoped flat and eeqaeb at xedueed peeeeufe being a eepty ovaporalor(at c 406C)- lift"And va(aw4 by WOi I mL of 27S wcabio add in mel0anel to iheper'16eped Oakad i1Yi,1110 Geii)dlel0ive, 7}aofw the sltrsb lo HPLC viW udnj diepoeebda pipea. A1ri1t" wu01a alai ca6oepeaY LCJMSM. 12.0 C3fOmilOOePby 12.1 I~etR fLe aeme amwtt af we1t elendaeQ, "load ImliEed ample wig thc LCJAplAi9 syetai A CaTi6ratian Wendcd mod preeedi nil Ec11oW all belyted t-VIN, 32.2 Shadar6e COMA eoerapandLy baletp Aveamen oonupnmion lovcb ' mawLe iocloded in aa anebliod eel. I1.) An mu"ne ew of ecQaavd ealibntion tendude mud be included at the be0innira aad at On pod of a anopls eae. Qx]eacepd apeadude tnuet be intaepaned belveen every S-10 yuoplee. As 1m altmneiive. an entiro SON nf eeRped eaKbntim atmleda guy be hpecled at tba bellinnwy of a td Oallowei by stuwted ca6bofian eleodftde iat.eqnned every 5-10 "Win {so aoamd 8n a xwa16 ed of athulW 111:11111130de). In aQur COW aM Iractwl ee0trafiun ahoda~e meet be the hit and lad Weolivn in aaample at 12.4 Um GneR "endud anrvea for qvamiledmt . Linav standeed wrven an Oeasaaa!!r t00 aia]Yre by Now rejretwloa oft IA weighting ofpeH; ores vans odbl+0iw eOmdWd aoaaakwgc wia0 MurL7au 3J (of opuivelcall eoAwen s)qas. 12 .3 SenpD eespaene abould ml eaeeed eOndaed r. esponses Any nrnpks that maeed Aafldad }aepooecs iLwW be fiatLer tlilWSd ad nanelynd. 13.0 Awepwwo aaeda 13 .1 ~ mWteAor ape~dl<deuar6s iptat 360 emu6omaparret 917413 rmn 7b 113 eau panot ounpoods to an "OAaoioe, ~hile am deollbler inn(!69 amu)mprumletbe loaofoabon d. ioxide rep sort /"' Pagr SQ oJ 65 Exygen Research Page 91 of 110 00011_4 Interim Report #23 - Analysis of Sediment Samples Exygen Study N.: P0000760 r- i:xygea Protocol Number: PON0760 fXyyisA~r~ Msdioil+~iY0W1'7)J ANA4Y17CAL 11Mpp i~bplil,AulydoOclhoDotwnonoWaotlRAae~oaaamaaAoid by LCJMM[9 A)iaVayemoca 13 .2 go" bLOU van mt aaonf0 "OA at bvdi 0 ~tv am do LOQ. H a bhalc ooaldw "OA N dbreY Vaow Cm 0.3 ppL, iEMa a new blwk uunpto marl bo obldwd ond IM aAin mt muRb >,extnwa0 . 13.3 liooormia ottamkellpNmr Yld mIWa "a MW be bmwaan 70-130% of Ow loMa vatow. If a **Awl 10Ite 6W aupldr the moePabla limits. lhe ~IYr rt efpmiplr dwub b~ nraxkabM . lUA My eaUhwdft 0064 bOOd lo Do a aolin9al a011iw by utirt an Huye Bnm Twt, my W qcludod dm da caiwlodon of ihs mdibuion curve. Uo'ravw, 1!o bW mlm6w of allkaliao obaderh dim soOW be excluded mwtaa aeeoW RWi otqual wunbwaFrtwndw6 nyemct 13.3 The oamolldM oa(11rirA pU ft t+L'bluiao MM lCMM14d mwt be i0.991 Oh 3a.p16b V nL'hrploe rerulq AI( oWidr MM lvoib. tl . ~0/th0~ rh'o mug be tdmn to s{fuK inWWmmR uperuiaq Mad the oUUdm6athAidprnt M otiuapla itloWd be raalymd. 13.6 RINK** liuw hlr++im'Mmdmdt wd mmplrn must not drift mom Wun . t~%w0ldOmao0ljdealnm. VnHOtimdmedeifleacaedtlhitnmiivitlvn rt wdylia) nmdna IM I* MW %09 M"3"& 14.0 Gkula(ow 19.1 ~M bN mIIpYY10`p11>1110Y 00 Ei1Q1Ylf td1~ qAD1WS 0~ PF~11 fOW10 (In n~mL. bnod on PHk aimJ Wag dw WmdW alt" (lioav trymwiou p~rtusl f~dbY7M Ff+Y 1.7qx mRw~ piOyYn : PPOA kW (op'mL) - fEYtY'!t_lOttwD ~tapo 1 ".4 Up 4w bilowWmpw8on co eaorwt qKaaoum ofPFOA foumd in nymL w MA (VA) . MQA fbaot (ypb)- nnrol. -2*W [YF=Illdm by .hleh 16 . emd+"otmn. wm:dilut.a, if neosm.ry. 14.3 Yer "wfflwYaliboa w(O Ymwn *wmmls of lPOA prim so vut.acuua me as followiq eqoodm b odhrLleUfspwomR rooovay. RD--70i1[ YW ~YbhltwmA{~~ . _WIY~fWnd iAC01md (ny>t3~ x 100 m~ytiroddKl(ay'p Pw 1 .0 Pcpr SI of6S Exygen Research Page 92 of 110 0 t) 41 a.1.IS Interim Report #23 - Analysis of Sediment Sarnpies Exygen Study No.: P0000760 Ecygen Protocol Number- P0000760 ANALYITCAL METfiOD u.u .ataume.e: v00011e5 - Iliaba~aorwc .l~rtOPISlsnru.Iwd..err~ra ..woctindcA~am'DA)LS.ua x . - Mluotmd 1Jvo b7 LClb~llifi Ae.btieal9lutift F.rr. ^ ' Exytp kwMh 3058 Rwroh Driv. NmCW1Lp,PA 16I1)t APpcwd 8y. Pal a ...ll7 Tabqi .I Lssdar, LC,M, 8qpn Aertrreh t41uf04 Due ~+"aDaft vice rrweeut. op.ruta.,, s,ry,e se~rh 101- Exygen Research TaW Papr. 7 Page 52 of 63 Page 93 of 110 000116 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: PDU40760 111, 8xygen Ptptocol Nlmlbes:P00007t10 r15"eultowrsp NrsWHwwiw Naool 7st ABOAi .Y1TCAL 1mrfHOq Sddhai odMdyrblx Iln Dile~itltllien afApilwtoecmaic Acid WOOk in Small Mu=d laveDy LClr!5W (_0 sow 76ie Ma" in Io1Le.emplpyed at the 1Lelqkp and yomlltuiost of perfluaooGpoie aspd by IfiliD PoAeotaeoe ilqwd Ckemampaphy ooppled to a tumdem Man dpsoua~Ris Daltotar{iaC1MSrM3)itt Nun artl1iver. 2.0 saw . . 2.1 AlrqeaEwer~aRleloeeotrptaetleee. 21 COMu11 Gu oppcupiqW id$Gi be0ow islqdlipi ac.y clatantcsl fer prapar ufety 3.0 9aepleftaqsiressaw 7.1 3.= ' 3.3 At West s 1 otteel u.ple lor aco.cdoa. Sampler should Le ytoetwd Were mnractien. Place do fiveea sampk in a food paoceteot and hontDJodn wilh ft ioe. Place Ox snapke in eomunur rd JOWL Open im Ana staaye orembylti to sltusr rev eehon dioaida wDlimsdpl. Seel end ybce do eYnpia io fROetD ttqsae until lime of YYlyeie. AIptNlely. $flrte is an 111sY8fcian nascent all'aample (-k&* Ow 7j~ dm onpreopeiej fe eteweryad Ihs saapk can be used as supoad. SeEb aollectioo pcoaedwes will bv epeolAed in the stmplinp pin ft ibis Pplok 4.0 AeepnfaeadSvrtdtsds 0.1 Walr-HP'LC wa6 as I4,deoa-ttrl,c a,ae 4.s Aoaadttlle-11Tl .cveds aA ABlntapiM Aelpte-A.CS.IIaMCsaa ".3 rmtlaaoeeueoieAdd-siSms-Awden s.o L~ntwt>avebnttne 3.1 A NO Pttktmess Sqsid dnnnsDoprqh cspebU of P-000 up is ; wolwnk sqoippei with avsiwbk valance injeotor capable ofirjectiq 1.200 PL 000neGed 1Oit08ft Mast IIpc.40mItaf U.CJM9IMS). 5.2 Ado"iesto onUsnt cw~dqafarpedlo inmynti0n Wd qoaottntien. S.] An1134Gi! 6eLoee ap" of reading Is CLWW1 S." so mLd6lpocaEbpslypsoPSleneceoxSlitjeto6w . !,S 1UJaLdiopxs61epo1ypopyiOwcaitrilL2stv6ea . D.! Dltpowbkmiatapdpel(SPIOOoL, IolY70l1uL}. sa 12syn,L LIVE .uro,r-meual eenlee. 311 Zol.w.rllpuviatxn. res. i.r Pagr 33 aJbS Exygen Research Page 94 of 110 000117 Interim Report #i23 - Analysis of Sediment Samples Exygen Study No.: P000076{1 /1 8xygpi Protocol Number. POODD76D ir.mrrwww vo6o1rta I4sGlwmeor~ao aftaDwraxwwic Add {CFOA) in Swall - 1.ivw 6y I,CA69,7M3 69 plqon16160043.18 AuaoDIp~Yt(SOF10DDyywdi0 "]OOyL1,w~W6Upvwbkipr. 3 .17 W11pScpTrcVw6x{1~+clearEew~aia,a . 5,12 Sr6ruam aidibld. S.11 15uucmirw. 5.14 1Kir4e6roa dab3-13 C.MaW rapbl.or*wAag uML pflyPooYl..n wua.r 3000 rpm. 6 .0 pasmi6w*ka S3we% Fl AiayRkA Coliaoa: PkoqLwRT{Kqydmw 9ci"Bc), 2.1 mm r SO mm, 5~ . (Ylll: DI503-0f7130) 6.2 71earuntlw: !0'C 6.3 16olGhPhcr (A): 2aJI Ammowium weeuee ie W~r 6.4 LtoLlk PAre (D) : Memrol AS (;1rdlad Prepsm: MpILIOW hA O.D 65 1 .0 47 9 .0 73 20A 73 2L3 65 Flaw Rue !S$ WA1i1! ]S 01.3 SS D .l 75 D .7 77 0.3 35 O.D 6.6 1cjeadun Yohmew: l5 pi {aa6~ mowed1o umuct ss 30 pL). 6.) QrmdndoK Tqtt Am1- wdaodaMniud uliLradoa nwe. GE RmTeor .13mie~Y4 7he Arm caqdtiew ww uaaded r a yuids ad my Dr abs4ai in m4x w opdmdmfMHncwt~ 7A Id8l1638yMm 7.1 b(aft 81ecRacpmY Ne.EWwAd(]N modc, momdYhiRq 413 -i 369 m% for 1FDti 1'1e Aots cwdllicot Am miatd r " Vide nd may be c6aa`ed in owdar w eptitiiutiw MSMS rynem. Pwld1 t1 Page 34 qj'65 Exygen Research Page 95 of 110 0OQ11_8 Interim Report #23 - Analysis of Sediment Samples Exygen Study No . : P00007b0 rk ExygenPPDmoflI rnober: roooo7a0 O51aw1Metw1 AwAIMM %rrgou Malbad ofAMai2dw fblalw IIt~edae af 1{aomi Iiver b7 LCJDIS+HS kMhad Nnnbrt VPODI7a7 Add t]'F~DA1 in Sxull i .0 AspeatfamofBaDail~na i.l Mnbr'IsRkav 8.1.1 Y tnlS simoaritr wWS 1a waEnt u DrePasd by addiaf D .SSa q af aernmdtraeehta to IM mL ofwataK . A1wna10 vslmM may be pnprad 9.0 EtaMNdlr"q$oR 9 .1 9tandrd StockArarGlratip] Solurion 9.1 .1 Ptwffa a wPCk edPtim of -10p P~mI. ofPPSJA by wciqlti n~ !0 mr . o(nWylipli_aqprUtd(opeweSedlbr'wlt~/anddilutetolODmLwitb mrlknmol in a 193"w1 . LDPE boRla 9.11 AlA 1+>hpL fotq6oaoun pLndpq of PFpA is prep" by bringing 1 ro1. of Ow IDO p&ML Pohttion lo a !laal vokunr of 100 with metlrnoi ho aUS AWL LDPZ beds. 9.1 .3 A 0.1 p&hnL. bqillastiso aPlxiemof PPI)A is DrWoW by briniini 10 aLofOr 1 .0 N#sL wlu6on In a Eml vohnne of 100 wilh rtrtbnot in al2SmL LpPBbastk. 9.1A The M* and Rtdl!rdan Polwiau an so bs nored in a nfa(qararor at sppraorEmNdy A'C sad an atabls for q muirPym pariod o! B monp+s 8nm t)ttrdua a8pepoHim 9.2 9faadod Cdilntiaa solutfomi 9.11 IdY!(8tdlS eallSncos standards an preDld in malluoal viadilution *fib* Ql DYIoL f1~stiEo~oq aoiutim. 9.2 .2 Tfir gol6winY ia a lypitd aaaotpk: additional coaoenbatioaa msY be paper" Y Oa011d. Cmmhmnn ofPwrilkaia VnWms awaamrnwl.l ~_ Dihr1Y tml.l Fnri Carcnvnron ~1,~ 100 fA SOP f.D Igo i.D 3PD I.D IDD IA t041 Lo 5,D to 100 D.s L0 to 1D0 P2 1.D 10 100 Al 9.23 Sku H aikeaor eMUlrda ia 1t5-ML LDPE naaow-mouth bodes et YC so 6'C, lip to ds mondu MA Alpera volwas end aomenpaliona of "a,d,ofa may bo Prawrod aa P404 .r? Exygen Research Page :'b of 110 Ofl-~~~19 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P(}000760 l'^ firtVaen Proloeol Number- POM760 asq~~nwreeb ua~ew,.arvooolra / . ' AMAi.17iCAL 1dEnipD itllhod efAn.lS~alhreM Dweml6~be etPoorflaostepate"Add (PPOA) in Small . Maomm.lllYarbyLCJMSA1fS 10.0 aelebbetilp MI 'Feob b.9Cb t,f .mqika axpuwf {c/0449y 20 ur Ness) mutt ipcle0e at feem most embattled CA" and low tuakaded ooeaalt Ibttiti .d au known vaoc~atieerpabav.hdi~'a) wnrdYD~sdyc+l rew+xS fbr Wc 61rtch. IOZ Requirwmb br6eld .qd IaboorshoryAeplicaon and apikaa wiN be tpuife.d loan "01YUMMMPIMINobillyntarL 1lA SraoLExRklioe 11.1 We* 1 j of epple iwlo o 30 mt pllqtopylen. oeanilirp tebu (fOrtity as n..A .d~ 7pl.es lid p1d t4iR well} Nare quf alternate weights of liver may be bwwfnd dayeedhy oe Or aeaple sin avellablelor wa . 11.2 AM wapelodanmybllrallodeeWtnootl0mL 11:3 Floanmpqifememleofinpatiwemlsrlar-1 ednum. 1L" TrwRr 1 ML of Iho nrople wing a dkponbk pdpecte into . 13 mL I I .S l1 .6 11,7 !IJ IL! 11.10 11.11 AM f mL of enEmiikib ala k+Ik far-2O mioelw on a wrifl-action Maker. CmPdllya do Who o-)000 Ira by -3 adnmaa Does* the .upauelaet into a!0 mL disposable tootrihgo tube and add 3S mLef ralr. C.bnditim doe Cu 818 cwlrid6ta (I S, 6 mL) by posting 10 mL neatbaql Jbllevodtiy S mLoftlrLC w .lw (- t dmpyuc) . Do nat lac eohmat ixn dry Lodmeea~Pkmeoedldm.dCuSPEoetidLo- Dierardelsaas. 8ima,witb "2 arL of medmel . OWGeot 2 ml, of dupe mw . .*rdened 15 mL polyprepyleaaom6iGqe blbe(dmal tnbnsme- 2=L). AeelyselrsplxeeineekcdoqYaylCIMSrMS . 12.0 Clqow .rograyby al 122 1] .3 22 .e Wool dm wpe uaowd of eWr d=6rd e.mplo and fmtifiod aamplc ma thc kCJM,4JIIS syoitebr. A evlbr.tian alleJuQ muat precede and folle .r all llotlY++d e-Vls. Standards 4fPPt1Aowrapeedla6 ID aQ JUNfive or pane conoenuahios larsb sm!be feeYeea in .. aeetytioal AM . Iut rrqks eet er o.llbnlfm tOndnde mata loclod W at ihebaibnmS aid a the ad of .simple .A. Sbutd.rde mug 1'e ial .rapar.ra between every s.l D Y7lplaa. I11 bkllCee4v.0 pt aellee aR of qlibtftiell JGOdaI4i way br kg.cbd r 1!a bepsuoj of a am followed by cdibntion awidardr inlmlpU+rd mrrs-lo emtpla [w .cconne fer..epone 1a ofilaaa.rd.l. In eYOtr are, eallOa.Uen tttaOnCe mug be IOo flrn aod Last injeciion in a ample ad. Use Ilomr ahedad eaevea AV qo2efibilkno. Lima standard coves, 6Cner.pdhe Ow auwiyee by Motorrapiepioausing Ilx wuyldizs ofpeYc MI, pop s 017 Page S6 of 63 Exygen Research ~ , ;, - Page 97 of 110 000120 Interim Report #23 -Analysis of Sediment Samples Exygen Study No.: P0000760 r ExyQan Ptnt00ol lVUfnlfei: P0000750 B"e1u.sl Itwhe114w6nvOGona I - ----AlLALYACAL lllS7NOb Mwhod ofAutpielbe AeDnmlwdeo o{Pwllnsroomppd Aold (PPOA) in Small IRmmW Liver by WNW$ vwrw eel6r>tinn Y=dPA ooaeeqtnRiOO wing L(wiLyrot 3.3 (or equivdem) l2S Sauryk nAanw Whould oat eaNd 3Innilnd naPonaw. Any oarnples rh.u aseeed rA~ds~pa4es i+wJd ba fuNw OLApa6 WA nrWysed. 13.0 Aaqhmarrflileaia 13 .1 CArowlopnm muM nlfow sywkof n6WP1iv ianat 369 mu knee a parem sf 413 aM 'YA~ ~13 anns preat eornRnol~ to pw PPpA anion, wlrilotiho dqqhlu No 069MM)repieKnp ft )owofeaiboo diwldo . 13 .1 Ifdlwd b1La1n niul net eoWmi PPDA at lmla Vaakr tbar Lhe LOO. if 6iant cautdBu PFOA at Ievdb ym" er L1nA 10 aft their a new blk* ample ma absi.ad mA me veft so mw be re-ex 133 1lnarwiw of aoalYOl MOM and mntrix rPilna anral be bxwaan 70-130% of Ihdr lmown "M If & e0enel 090 We nreW On wu7ubbi Nn*" the Ktln wt of wOrp)w dpOk bo twakaepd. Any pawlx Van uuuida 70- " ' 130% a1on14 lta ardualbd by $e andyat No dAatnine if malntviua is ~mrlmod. 13A Any ealbwlioa rMOdrd Mind to be a wind" ovllia 6y ruin` lpe Huge Below 7nf, n" hr, wmlo" bas tIM ealwlulee of tha nilKadon curve. Ho7rwu, Go WW emhw of OdIhO00 Ipuadwh Iet oauld be aealuded anuxnel asoad70SLsdS1wIoW rurabR of apodar* h. owled 13.7 IV pawhalon cwfidaot (it) for alibntioa ewws lamented oqut 6e t ."02 dtr ZD.9RS). I[ whbralian enulu dll am" Ihna limitt. oim qqlqnilfp Shop WA a Ukm ta a41aa foWupoan oyir>tiui and dc mediodoallao nltr~ul aewmphsalwnW ho rwuiyma. 13A teeruio. dews iao.aea md.a. rrd .+mple, maw ro, drift more ar, t 1'~i wi(~e manalltical wn [i rswnflm fine ddlt excaws this linrt wilhm u auMial ~ lbatha wt muu brr rwW]~sd. N.0 Gllarlrion 144 Um as f6doNA1ioP eqqela0lo eWindslo TAe amoma of PFOA lbwid {in nymL lowan Pnlt NO) Lviuy the UWdaid currs (know regression p"Metcw prrmld by doe Efas Llmz ioflaui pmjraw: PFOAlpwd t0mL) - OnLam-Meerewa~ x CP x d4wr fmar a" t*-flow bywhiahfhn firul rolnme rn dilulsd, if aaesneyAiqwlfoam -10 pop 6 .17 Page 57 ofss Exygen Research Page 9$ of 110 000121~ Interim Report #23 - Analysis of Sediment Samples Exygen Study No . : P0000760 ExyIIem Pfotoool NWrober. PDDD0760 urr+zm-wk 1fefted Iumlrilsbr wa"a..eK VoooIrs ANALY'i1CAL AET80D ca 01PROwoockmak Add "OA) in Small _ Maima Cirw by LGMSMS 34.2 IM Ympln b11f6W WO lmawn mfolmlr of PPOA pdar to ernmpqpn, vu 11~ bAY+'ei 49rat1tN1 to a)cdpv Ibe pttcsAhwrwy. RC00Ory ('K) - , [ mhl mJYw romd(p-ilmYLw)a-daune~ibaWmfoLr)od in eouerol (ab'+nW) 100 1~.1 Uh r6r fo0o.wiyeqoatiam w ooavsr the ~onum af PFDA found in nymL ~o +qr'i ~PV~), r_ tUA MW pD) - A'Faut 1Qyaagap T-i x nn j reluma rmLll --pl+waib iW Exygen Research ~~i ? . .(v ,7 Page 58 of65 Page 99 of 110 00W0122 Interim Report #23 - Analysis of Sediment Samples Exygen Study No, : P0000'7b0 Exyyea Pro0oool Nucqbpr : M=-]W AWALYMCAL METHOD Ideahoa rYneber. vppol t86 HAW of Arab+b Ler Ih DWrab+rior d heAeawacUUSPo Add (RPOA)1h sm.n MadmN ~enw bt LC1MBlMi ANWI'tkWIaceXFWiGty: Brypnkaesuab 3058 Reaeenb Drive sowCONW, PA 0801 Appmvw Dr. .. Ywol Due 7ae)mleal Leader. LGhli, Faqpa $eecegtE Exygen Research row tqe.: '+ Pw ss ofas Page .] 00 of I 10 0001.23 Interim Report #23 - Analysis of Sediment Samples Exygen Study No. : POa00760 /" Fxyacn riotocol Number: P0000760 Rrrrth ia'Aodyiir IiwblMuuEer Yuoot7ae AAALYIYCAL METHOD . ~ ~ slfer~io AM (PYOA) in Small Mmued'aum by LGhLSIM3 1.0 Socpe This mcWod ia to be wtqtbyad for Ila lsolstion aad t;uautlmion of partlwrooataeoic acid by Hlas Tar7tQUaooe Lfqvid Cbromaoosuip3y oaaplad lp a Widem Mass Spaoreomeoao DemNtor{LGA191M9) in =all mammd astm 3.0 Salby Ll AlwayaobKriadrhbordaaypra .9lca . 22 Con" 11w tprcaprlpc MSDS bslbrs 1aNliquany ehaMc:I IbrPwpDr safay V011-ill- 3.0 SaeRla Aaqolraml ,. ; 3.1 Ac LW 1 ML Of "d aa,apia far e:Lacooo. 3.2 No iaffilt4 pmsoa's,0 la owbd firaanmr aataplw. Ho~ar. Gom serum aao*la AM 10 allowqdlo Cqapblalyfhaw to tenm tavparuna bafae use. 3.3 immp7a wIIn:oen prowduw will bo qmcikcd in do uMlics plat fm this w0 It.apeu aaa AUedanh 1.1 Waw-BpLC8aW 4,2 . Meqqmot-Nl601pwia 43 AulmitAla-HPLCSade 4A Amaarinm Acdtw-A.C .S . R,wpat (3tada 4.S Arfiweooa.looieAcid-Sitma"AiOrioA S_9 Iaa4ooaat aed Qqaipneat 7 .1 A WO petOxsfea liquid eMempoprWh olpabk of puoqlny rp to ioWSrti aqdppad t+'l1Y a WW" >oluun iqjxtor oipaWs of iqjecuttS 3" ZIIP KL soeaward ID a Wrdwu Man gpaVatume (1.C1ARSlMS), f .7 A dsvia hr sOUWOt Ow 4Wlbr DntSntepll4ipn wd qtwuintVon . 3 .1 AWyNaI baLnee oapi4le uf eudiosto 0.00001 S . SA 50 aS dipeplb yolyDroPyNa unei6qe lobes. 3 .3 lS ML &IpmWA pol.YptaplAaw anbtlltae blboa 7 .a D*paaablsadwopipata{SO.f00aL19(M1LOuL). S .7 12S.mL LION oorew4bonR bNCO. SA 2nl . ekWMIXrW lut S.9 niRoaabhPipdler. 3 .10 Autoptpaen t100-1400 ILL md 1a10o pL.L with dia9a "ab1, tiyr . 5 .11 wae .a Sapr.ic We a ac (IS) Icli PO mtr.aW. alu Srs .a.wmaeanirola f.13 VaaxR. Pqe 2 .(, r' Page 60 af6S Exygen Research Page 101 of 110 Interim Report #23 - Analysis of Sediment Samples Exygen Study No.: P0000760 Exyael P=ocwai r),nnee- poooo760 hyP. how* eiv:a.rncu. METH.09 Meda.14.N.r y0C0Mn MamodorMd7lsis.lb[.tl7eDdon~io~Aion ofiWlluaooetuwie Arid (PApA) in sm&il Mend Sauma by LCJM6+RMS S.i4 W~LL~dSm~Iflur. 5. 15 Ccultilbp sip" dapianing i! omL paiyyroyylane mLea a13000 rpm. a.o chMMwsw+we sywm 6.1MYy11W Ooiuroe:Fiuepbw RP (1Gypon~ Sciatti&), t.l mm a 30 nm . SN (PJN:8230'F0621~) gl Tam~amxc 30'C 6.3 Mob~lsPnew (L) : S mM Ammonium Acatia in Water gA AIdMbPNw(e) : MOWMl dS Cradimtl4agram: ` 1'Loafnin~ 2iA QO SS 1.0 53 i.0 23 20.0 25 22.5 63 Flovr Rut }Fi ImLlaig1 35 0.3 33 0.3 IS 0.3 73 0.3 33 0.3 0.0 *onion Yolume: i5 KL (cmboinaouedlo a muchu SO pL). 6.7 Quaotikotioo: Pphluaa--ncfaad Mabdard e+libraeian cum. 6.9 RmnTimr-23minvMa. Thsatom a"upes areinomdnd n*guide andmqrbpduogadin nvdarto aPtimir Iha H7LG arlm 7A MSMSSypwp 7.1 AIadK $IaamplpayHepalLviblABd moab,muniLuring4l7 ~ 369 M'S fat PI" The abore condhipw are iauoded a a pude MW may be chimsed in oMnr a1 oPdimMths x3115 ayauM 2.0 RontiooofSahnima 4.1 Mebib PMa R1 .1 2 a~M ammquiu~a uwpa is rna i1 ~paesd by s8iias 0.1 Sa j of irnaonian sc .aaio 100amL ur+ruit . Aroaow vohmw my be pmpaw. Pop ~ -r- r+ Pap 61 of 63 Exygen Research Page 102 of 110 (30U125 Interim Report #23 - Analysis of Sediment Samples p. 103 Exygen Study No. : P0000760 r 8xygen Protocol Numbey: P0000760 a,ya Irra .ce . _ nfaw rw.e. voo0te ANA1.mcA1. Msraae UdW Dr Aadyw lb did, DelamsbUal ofAerllwrovelanoic Acid (PF A) in maD 1 e- rmlat Swim by LC/M&MS 9.0 Swndacd PnpwaWa 9 .1 Suylud StnilFarliBalioq 861alion 9:I.t Pfrpri a rank roAttio of-100 pyht of PFOA by wtijpiS i Ding of malytical a-dwA (m-?WW !oa pqpfry) mad dilais lo log ML a.nh moshavOl is a 1. 1S-al LDP80atla. 9.1 .2 A 1 .0 ItWmLfurd6oYion qAition of PFOA ii prapus0 by bringing I . ML oftb 10.0py's[ . outinn in 61ina1 vol4mna of 100with meDnnor in s 12f s1L LDPE bcnla 9.1 .) . A Rl W+mL Ioetibwlioe eelntianolPFDA is prepard by bringing 10 sJ. 0 11114 IA 1troL aolume W a final 1; 091m4 of 100 whll mnrunol in a 123 esL LDPEbot<k. 9.1 .4 9b dad[ and (otCdoabmt w1uliooa art Io ba aEarad in areBit;or6lDr at apymainuply4'C and as atabk faramaximun pvlod or6 monCv lime dn: drmeefpaplprAfa. 9.2 Standard C6libntiem Sobulorr 43.1 LGDd91WLaJikatiOaWadqdtp6PnpRbdinplelbanalviadiiutim of9uQ1 p~lalLlbplilicitiom aolwim 9.2_2 76s fblbwlltg is alypibfl example: admrlrl eaoomtratis may be pnrgall Yno" CWACAWOK NFiw~illeauon so, (.I-,, 100 lOD L00 S.0 2.0 Vowvx jjO 20 Jill t9 IO t>",rI.~ to LOO MD 100 100 100 Final Caianbuia uwmt.l .01 12.U IV, O.S 0.2 91.3 9.].4 1 .0 16 100 0.1 3.k eii 125-ML f.ApE,qero-uwuh bouka at 2 "C to 6'C, ny to Yx ansth. Allwmol6 volnmrs aod mooqmertlorq of au,pdw<dt meybt propved aa "kW. 1910 Dwh 5M UP 10.1 P.wb board, or wmylar Qeae4d (typleally 20 or laay) fokw bCIuQa at leut am wlt;ppd pOppol and two in6anad oomoola llxtilSnd at known wneaotradma (lab csntrol ipOcs) to "wify poeedmdrecovery far tke Ebtab. I D.2 Rsqlirclaai4 far Said add 11botamrydllPlluta ad spikes witt be ~paiDaf In rLr qoaliprarwraaos PLO R7 two ptojrtt. PAP- .I J Pop 62 of65 Exygen Research .. `, Page 103 of 110 OOOI+zs Interim Report #f23 - Analysis of Sediment Samples Exygen Study No .: P0000760 Exygea ProtOapl NwnbOt: P0000760 ,yrasw..nci Af&l1.YTICAL METHOD Dlv~dOfAoaiydtlbrlltDinambitilloa Ofla lrlapwad 9m" Ly LCIMSlMB wya N,.t, vooot~~a ' Acid (PP9A) in5mall 11 .D Swp1a Sxaaelam 11 .1 A(aura 1 eI. of sample lift a 50 mL polyptvpyle0e eeaailpp tuho (feeify q giant oeplh0a !id mind nut wdl). ?lok NW aftpraaft vola0tat Of asnon sfyl b'mewni drpandft on IM sample fka avwflabls fer not 112 A6dWVNr001hawdtplOfOf~GWvD1w00af2De1L CIDtIpAlly t L3 vo<a .hr-i Mind l1 ." llaeahc 1 t.ut Of 1W csnpk vwbj a d4ponbb pipana into a IS mL iirpaabfe wmylr6a 41011141, I1 .S AddSml .atamlenit,ile ad shalom fm-20ainnua son a wriat.action shata . 11 .f 10 11 .9 ' 1L! 11.15 11.11 L1mIa1blp4a "asat-3000 toofOr-5 embtnla Daa'aot liawptaaxomat into a30 mL diopaaahla eankihp whit and %ft 35 mL ofwatar. Oust" 16p Cu 6PB aa[hidaaa (1 p, 6 ivL) by passing 10 ML MaLbsivol 1h0owadby 3 mL ol1>r1.CwalOrt~2 dteprwc} bo aot lq coiunn tun dry Load go asiiDlt oasaadltioael Cu SI"E caatri~a, Oabcand eluue. &~]q w9111-9 roL of aaahaeol . COW 7 !rat eLUla imo a pnduatcd l5 ml .polyprapylaw eentrilhpp tWs (NW vaitw- 2 mL1_ Analyze II using witoubcopimyLCMINS. 12 .0 ckwamxtvfmft 121 LjaR tha an= amo0at ofacb plrdatd, sraplv AM 6otlfAtd toopla inu tde r` LanCaIlMySsItDLn4owssysvG,an . A cauleadrqm v0atdatd mud paceds and follow all 123 Stadacda nfPRQiA toIrGpOfdlnQ f0 N WAR 8v# of own o00emtrazim levels vow OebdYdod i0 its mlf6aal rat. 1xJ An eduaeam"ofsaUWadOnwndWlrawtboi0cludad+Otbeb0Yimi']ing anOrt tTaasdofasuopk vk. 6Yrrdaada arwt be inter~poeiod hstwaa rrecy sIn annyla . ila an ahernatira~ an endee as; of plZnlioa atandarth may bc iajqeeed at the bKiaelat of a in tonsured by cawmatmm standards ioterrpcay rrwy 3.10arrsplar (to woo" %braaeoond rei of awrdarda). In silhar Cast, wlibatioo paco7atda MM le the t1n1 and last injccuun in a noaDla ast. L2A '1w Imer Mamdad ewvaa Dr quaatiud0n . Limn ala0dra curves am genaraud 11w Iha nrlKo by liaweege,aaioo using !ht waightin` or peak am rwaua calibralioe alaaaatd ooaiolattntion ueiiig Mwl .ynz 3~3 (w equrvalvnt) wlh+vn Stations. 12.3 Saint 7ecpOnp aboiiid tlot cxceed aundatd tesponmew . Aay wnples that acrd wrdaad nrpoawaa abaWd be Batlxrdilondand ranalysod, tap ! or' Page 63 Of6i Exygen Research Page 104 of 110 000127 Interim Report #23 -" Analysis of Sediment Samples Exygen Study No. : POOUa7b4 I- 8xyyea Protoool Numbcr: P0000? 60 6&rani+ewn4 MOooaNea6n voao1:N ' ~N~4L.lEI1CAL " DlILTFW9 =MYym1br ~}ehrmYtmor~ a Palledmosevwic Acid (PFOA) in Smell .. . - .. (2attand Swam by LVM&%s 13.0 Aeaaplrro L1iPa1L , 13.1 13 .2, . . 13.3 chnsomociinis aria nlow a pah Of e danypaa ien a1369 pass Bum e parem of4,13 aolu" Thr 413 au praat oarrpordr lo on FFOA eniaa wnile the dapFldr)wi769.a11 rrvnnea au lo . croueea dinsidR UeAx# Missile must, mat ood= IFGA et Isrell poster am as IJOQ- if n O*ll aonb(al; rFDA rk 1ai11W Peaks, IAsA 10 WiwL, Man e new 6fc* sapie ourt ba obWsrd md 1MwpiKawt mwl brastrxtW. Romemir of oaa~Yol ~7or and wwa qdua muw be Dnwwn TO"qOK of Hria 1>+awa valrra . ,I_fa ceo6d Van Wits *abide do aoerptuble liraiu, the 129 ".13091 a4ad~ I eraluaW by On MW)* wlniplrd MIWX d~tamime i~n:exuletiap~~s 13A Any ceGbr+dow atmdard lbun6 fo be s wtqtiav ordla by usoq din H,qo Raw Too, maybe wWoded boo IdncalerlLfioe of the tnlibnliae emvr . However, 1M ICQiI . YY0140r of eablistoloi4 MoodYdf 60 wald 11e eKe4u10d nor6aol OasAd 20KOf I4atutal We4saifaemdl0da 4ihtnd. 133 21e oomw]eiaa ooedlkiont (R) far aah" bmlop turves sannraed MUM be 2p997 (R= 2D.4RS), If w86qaim rKU14 AU ouwida these limits. then aqpoql(sAa rlrQe seat he taken to sijuR feytrmeyeet opesatioa . sad d~ s0uldrrda or 1M taiwut sel a~aol~]n ahwld be munlYeed, 134 Rwamiun tlma 6dwewt afllndaodl sea waplea rnmuw am drill male 12nn ' Y" "(" wltLin aa rnlYliu) flim. Ilr(aentlen lima dri$ etceeda this limit wilhin M 'aneiylioil AM lhro the h! Boost be reastrmsd- 14 .4 Cabolulcro 19.1 Uaa dr,fdbwl4pwGoa q qkuWo Cq Ilmaupl of lPOA fwnd (in n~hnL hard an ywlc ra) orintll IhM etaaAad cur" (linav rarnnian Farnmmsnl poratad by 04 MM Lps eoflwus prorm: FFOA folnd (sy1mL)- Maui am . inmccuq x dFx diquet hCwr slo" bF - Itirforly whlththe 16u1 valurne Who diAued~ ifmcawY. Aliqnol 1Yacar - 20 1a.2 Fa wbFbc farYSed wit lmenm sonowaet of PPpA prior to exlnction, ux do 16110frin/ aq0alioq to aNoulYe IL percent roeovtay- RXO-7 M - ~ WW tw1~Reload(ni~~ " ~Sf+~md mauouoll~L)~. r l00 ' ~rI,LMaddIdftWmL1 r.p s ~r " Pnge 64 o,t6S Exygen Research Page 105 of 110 000128 Interim Report #23 - Analysis of Sediment Samples N; Exygen Study No . : POfl00760 Hxygeo Protocol Number : P00007b0 eqym Lu=t wmeaHd .,voaonn A(rw1.Y7JCAL btiTBQD 1(r1Eod ofAodyws FMn'i0 DOWNEMmden u ropc4tqic Amd . Meeuael Sattm by I17MlAdS F A) in Smell 14J Up Ort l6llowiuj oqv0on 1n oaarat ihr . al PFOA fauod in n=ImL to gpk . "OA *WW (1T~~~ (tpAA ...1( .h. .T}1 tMiph sW~m~s {m4) jw" Exygen Research pop 7or7 Page 65 of63 Page 106 of 1 10 000129 Interim Report #23 - Analysis of Sediment Samples Exygen Study No. : P04U0760 ~~~i /RI,~~E~I-5EARCF! 3058 Rasaash Drive State College, PA 1619D1 Phone: ai{-272-1d34 Fax: 814231-15W i,iQMirQ_ . M . Qi ' Arnandment Number.j- FIfeaUYa DOW. Qjj191~ Exygan Study Number: PQ001 i31 Cilerrl Study Nnmbar. Pawl of 1 NoM DESCRII?EIOUF aMMu SECTION - ---- 1) Aaiely6cal Procadun eurnmary V000178D:Sa#1on 8.1 2) VeriBCatbn of Ara?AW Procedure AMENQIRDTO 1) Add W Ssction 9-l : Section 9 .1 .8, Altsmal9 wsiphts ad standaNs maybe coed to prepare altarnals concentraU4ns ad stock solutions as necessary. Alternate Web of fortllicatitm solutions may also be prepared, 2) Low and high apiktnp levels of the analy6es for each matrix mey 6e altered depending on sample shce avaltabfe for exfrac5qn and/or to cow anslyto concentrations 8)peated In to sarnples. 1) Hphercontsntratbns of standanisneed to be jonpArad In orderto spike to simple battles st hipher lemls. 2) The sanple sias available foran*[ rtarrnial barandsertxn was smaller dsan a"actad. Soft at !he pre-delorrillned levels In the protocol puts the spftd concentration km than the datacllon Ifrttit. Also, ft arm" levels In the ground water samples am expected 10 graatfy wooed the prs,dstarminad spiking ienble listed In Ow proloool. When the baels In ft samples Wpatly eoroeed the spiking (aveia, an accurate recovery vakie carw'yvt be calculated for dw QC aarnpis . Higher spiklrV levels in tha bottles WO cover Ow analyte cortcer+fratians opsded in the water samples . The LOQ Is 100 nglg for a 0 .1 g sample d small ~mammal liver and is 1000 ng&nL for a 0 .01 mL sangla of small manxrsal sornua, Higher lenels of :pBdng for the water eanipies wit ensure that more QC recovery data can be used. . r w, Sponr r nnrlnti (B "Mold) DoW "an OW PAWI&W ads? t19WqRYID : VV000122d-B= ADAYHISTiiATNE FORM Exygen Research Page 107 of 110 000130 Interim Report #23 - Analysis of Sediment Samples Exygen Study No, : P000U7b0 3058 Research Drive Phone. 814-272-1039 State College, PA i"0c Fax: ai4-231-150 Amerldment Numbr. Effedlve Data Exygen Study Number PROTOCOL AMENDMENT 2 0'.i107106 PQbt}7131 Gliant Study Nurnber N 1) RepmL page 11 of 8!9 2) Teal Mateliala, page B d 85: PF4S transition menflored 499 so. page 1 or 1 None AMENDED 1) Insteed of one flrlat taport, fnterim mpar>ot vw1U be issued. 2} PFQS hansflinn monftarod mey also be 499-~ 80. . RsrAsla7wfcaS:W~i1w~ 1) Due b 1he eXoessive oines of ft data gets, inEarim reporb rrDl be issued tv ellow ft dbM b oeceire data in s tirnellsr*rrwR omaur oskls 4*9 2) The API +fOW LC.IM.SAriS syatsrn! deReqt 1na 40 -> 80 PFCIS baumltlon Wih greater sarl:lldvlly thnn the 498-)- 991raneiGan. IMPACT ON S_,JUDY 1) 711e cYent will be able to reeeNm and review ft data mm Auia*" 2) The 499 -> SO tremition con be detaatsd with gresier aensifjvily; 48srefrxe, 9fvlng i #letber thrflcrlWWaphy. : LAPt 8k+dy ota~Ern ~re Do w , S Dow Lr . shwNoeIyrl'~ C a~wpnr++r+t~+v~ EXAM Lip x7~G1-f! ~~ 8poryrx 81' ~C _ ~ l +wH~-c~' 3' li Q~ s. . . MARY Vk VODC122O.a Exygen ReseaYCh ADYfNIS'*PLA'RYE FORM Page 108 of 110 000131 Interim Report #23 - Analysis of Sediment Samples Exygen Study No . : P0000760 RESEAttCH 3058 Research Drive State College, PA 16801 Phone: 814-272-i039 Fax: 814"231-1580 DEVIATION FORM Genaral : k Project Specific Deviation - Facility Deviation Page i of 2 Date of Occurrence, 04f2t3106 Exygen Project tt : P78DIP1131 ' Deviation 5 Client Project # : NA Reference # : 06-078 Regulatory Rrhrer. Deviation Twe: (Include Y# for mefhods and SOPs) GMP X Protocol Method 50P X {;LP Other W: NA None Notebook reference: NA Sample: Descriotfan : Login # : LO008181 Container # : NA Lot # : NA Summary of Revistf6n : The three sedimernt samples In L8191 (C0172892 - C0172894) wene,originaliy extracted using the sediment method V040,1782 : Poor recoveries were obtained for Pt=O3, PFOA and jC PFOA. Because of this, the study sponsor requested the use of an alternative axtraction for these compounds, as follows: Direct Injection Method; 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-mffliliter centrifuge tube for the extraction. Ten mdlifiters of 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 -3Q00 rpm . Each sample was analyzed by LClMSJMS electroapray. Using this method acceptable data was obtainedtar PFOS, but the .recoverFes for PFOA and '3C PFQA were still poor. Another-aftamative method was then used for PFOA and 1C PFOA, as follows ; Alternative 8PE Method: The samples that were prepared in 196 acetic acid for the dlrect injection method were used for this, ~exlraction . Five milliliters of each sample was aliquoted ft a 60-ml. polypropyfene centrifuge tube and the volume was taken b 40 mL with water. The aampies were then ceritri(uged for -10 minutes at -~3000 rpm. The supernatani was then loaded onto a C SPEcariridgs conditioned with 10 ml. of methanol and 5,mL of water. The eluala was discarded . Approximately five milliliters of methanol was added to the cartridge- Five mHllilters of eluate was collected into a graduated 15 mL polypropylene centrifuge tube. Each sample was analyzed by LCIMSIMS elecbrosprey. Causr : ~ Preparation _ Anaiysis _ Instrument ~ Client Request X Other tmtsact; More usable data was obtained. LIBRARY ID : VD001B4M t;xygen Research ADMINISTRATIVE FORM page lU1 ot I to OOQ1 .'~2 Interim Report #23 - Analysis of Sediment Samples CHEM EHSR 236 1B 651 733 1958 , cMr-1001 Qt:o1pa Fru-NM Saurlaa Exygen Study No.: P0000760 0+5101 '04 73 :46 ki-0Q.a;106r.o0n3ro1c093 F-91a Exygen Research Page 110 of 110 o001,33