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INTERIM REPORT # 10 - Analysis of Fish and Clam Samples STUDY TITLE Analysis of Perflnorobutanesulfonata (PFBS), Perfluorohexanesulfonate (PFHS), and Perflnorooctanesulfonate (PFOS) in Water, Soil, Sediment, Fish, Clams, Vegetation, Small Mammal Liver and Small Mammal Serum Using LC/MS/MS for the 3M Decatur Monitoring Program DATA REQUIREMENTS EPA TSCA Good Laboratory Practice Standards 40 CFR 792 STUDY DIRECTOR Jaisimha Kesari P.E., DEE Weston Solutions, Inc. 1400 Weston Way West Chester, PA 19380 Phone: 610-701-3761 INTERIM REPORT COMPLETION DATE May 09, 2006 PERFORMING LABORATORY Exygen Research 3058 Research Drive State College, PA 16801 Phone:814-272-1039 STUDY SPONSOR 3M Company 3M Building 0236-01-B-10 St. Paul, MN 55144 Phone: 651-733-6374 PROJECT Protocol Number: P001131 Exygen Study Number: P01131 Total Pages: 132 Interim Report #10-Analysis of Fish and Clam Samples Exygen Study No. : POOO1131 GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT Exygen Study Number P0001131, entitled "Analysis o f Perfluorobutanesulfonate (PFBS), Perfluorohexanesulfonate (PFHS), and Perfluorooctanesulfonate (PFOS) in Water, Soil, Sediment, Fish, Clams, Vegetation, Small Mammal Liver and Small Mammal Serum Using LC/MS/MS for the 3M Decatur Monitoring Program," conducted for 3M Company, is being performed in compliance with EPA TSCA Good Laboratory Practice Standards 40 CFR 792 by Exygen Research. / d John Flaherty hincipal Investigator Exygen Research / Weston Solutions, Inc. Michael A. Santoro Sponsor Representative 3M Company Exygen Research Date Date Page 2 of 132 Interim Report #10-Analysis of Fish and Clam Samples Exygen Study No.: P0001131 QUALITY ASSURANCE STATEMENT Exygen Research's Quality Assurance Unit reviewed Exygen Study Number P0001131, entitled, "Analysis o f Perfluorobutanesulfonate (PFBS), Perfluorohexanesulfonate (PFHS), and Perfiuorooctanesulfonate (PFOS) 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 phases1were 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 Insnected Date Reported to Date Reported to Principal Exygen Investigator Management Date Reported to Study Director . Raw Data Review and Interim Analytical Report Review . Raw Data Review and Interim Analytical Report Review 08/01-05/05 05/03/06 09/20/05 05/08/06 09/21/05 05/09/06 09/21/05 05/09/06 !Note: All in-lab inspections will be documented in the QA statement for the final analytical report at the conclusion o f the study. This QA statement involves only the review of the Interim report and associated raw data. Exygen Research Page 3 of 132 Interim Report #IO-Analysis of Fish and Clam Samples Exygen Study No.: P0001131 CERTIFICATION OF AUTHENTICITY This interim report, for Exygen Study Number P0001131, is a true and complete representation o f the raw data. Submitted by: Exygen Research 3058 Research Drive State College, PA 16801 (814) 272-1039 Principal Investigator, Exygen: Date Exygen Research Facility Management: Zco6 Date Study Director, Weston Solutions, Inc. {/7/J Jaisimha Kesan P.E., DEE Weston Solutions, Inc. Sponsor Representative, 3M Company: Michael A. Santoro Director of Regulatory Affairs Exygen Research Date Page 4 of 132 interim Report #10-Analysis of Fish and Clam Samples Exygen Study No*; P0001131 STUDY IDENTIFICATION Analysis of Perfluorobutanesulfonate (PFBS), Perfluorohexanesulfonate (PFHS), and Perfluorooctanesulfonate (PFOS) in Water, Soil, Sediment, Fish, Clams, Vegetation, Small Mammal Liver and Small Mammal Serum Using LC/MS/MS for the 3M Decatur Monitoring Program PROTOCOL NUMBER: P 0 0 0 1 131 EXYGEN STUDY NUMBER: P0001131 TYPE OF STUDY: Residue SAMPLE MATRIX: Fish and Clams TEST SUBSTANCE: Perfluorobutanesulfonate (PFBS), Perfluorohexanesulfonate (PFHS), and Perfluorooctanesulfonate (PFOS) SPONSOR: 3M Company 3M Building 0236-01-B~10 S t 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 1IME i ABLE: Study Initiation Date: 11/05/04 Interim Analytical Start Date: 12/27/04 Interim Analytical Termination Date: 04/17/06 Interim Report Completion Date: 05/09/06 Exygen Research Page 5 of 132 interim Report #10-Analysis of Fish and Clam Samples Exygen Study No.: P0001131 PROJECT PERSONNEL The Study Director for this project is Jaisimha Kesari at Weston Solutions, Inc. The following personnel from Exygen Research were associated with various phases o f this interim portion o f the study: Name John Flaherty Karen Risha Paul Connolly Amy Sheehan Christine Edwards Brittany Kravets Mindy Cressley Krista Gallant Scott Crain Eric Hoover Carlyle Morrell Shawn Robb Mark Ammerman Eric Edwards Edward Cams Title Vice President Laboratory Supervisor Technical Lead-LC/MS Associate Scientist Technician Technician Technician Technician Technician Technician Intern Technician Technical Lead - Facilities Sample Custodian Sample Custodian Sample Custodian Exygen Research Page 6 of 132 Interim Report #10-Analysis o f Fish and Clam Samples Exygen Study No. : P0001131 TABLE OF CONTENTS Page TITLE PA G E ................ ........................................ ................................ .....................................1 GOOD LABORATORY PRACTICE COMPLIANCE STATEM ENT.............................. .2 QUALITY ASSURANCE STATEM ENT.................. 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............. ......................................... ............................................................. ...II 2.0 OBJECTIVE..................................... ........ .................................................... ....................12 3.0 INTRODUCTION................................................... ............................................. ............ 13 4.0 ANALYTICAL TEST SAMPLES.............. ............................................................... .13 5.0 REFERENCE M ATERIAL....................................................................... 13 6.0 DESCRIPTION OF ANALYTICAL M ETHOD............................................................15 6.1 Extraction Procedure............................... ............. ........................................................15 6.1.1 Sample Preparation............................................................. ....................... ....15 6.1.2 Glassware Preparation............................................ ..15 6.1.3 Column Preparation.............................................. 15 6.1.4 Sample Extraction..... ......................................................................................15 6.2 Preparation o f Standards and Fortification Solutions..................................................16 6.3 Chromatography.............................................................................. ......... ....... ............. 17 6.4 Instrument Sensitivity.............................................................................. 17 6.5 Description o f LC/MS/MS Instrument and Operating Conditions............................17 6.6 Quantitation and Example Calculation..................... .............................................. .....18 7.0 EXPERIMENTAL D ESIG N ............................. ......................... ....................................20 8.0 RESULTS................................. ............. ........... ................... .......................................... 20 9.0 CONCLUSIONS................... ................................................................................ ........... 21 10.0 RETENTION OF DATA AND SAMPLES ....... ......... ...................... ........................ .21 Exygen Research Page 7 of 132 Interim Report #10-Analysis of Fish and Clam Samples Exygen Study No.: P0001131 LIST OF TABLES Page Table L Summary of PFBS, PFHS and PFOS in Fish Samples......................................... 23 Table II. Summary o f PFOS in Re-extracted Fish Samples................................................ 30 Table III. Summary o f PFBS, PFHS and PFOS in Clam Sam ples...................... ..32 Table IV. Matrix Spike Recovery PFBS, PFHS and PFOS in Fish Sam ples...................33 Table V. Matrix Spike Recovery of PFOS in Re-extracted Fish Samples................. 46 Table VI. Matrix Spike Recovery PFBS, PFHS and PFOS in Clam Samples...................49 Exygen Research Page 8 of 132 Interim Report #10-Analysis of Fish and Clam Samples Exygen Study No. : P0001131 Figure 1. LIST OF FIGURES Page Typical Calibration Curve for PFBS in M ethanol........................................... 51 Figure 2. Non-Extracted Standards o f PFBS in Methanol, 0.0005 pg/mL and 0.001 pg/mL, Respectively............................................................ ....... .............52 Figure 3. PFBS in Control Fish Blank, 0.00625 pg/mL Fortified Control Fish Spk A, and 0.025 pg/mL Fortified Control Fish Spk B, Respectively........... ....................53 Figure 4. Chromatogram Representing a Fish Sample Analyzed for PFBS (Exygen ID: C0049478, Data Set:061605E)................... .......... ...................... 54 Figure 5. Typical Calibration Curve for PFHS in M ethanol.......... .................................. 55 Figure 6. Non-Extracted Standards o f PFHS in Methanol, 0.0005 pg/mL and 0.001 pg/mL, Respectively................................................................... ...............56 Figure 7. PFHS in Control Fish Blank, 0.00625 pg/mL Fortified Control Fish Spk A, and 0.025 pg/mL Fortified Control Fish Spk B, Respectively.................. ......... ...57 Figure 8. Chromatogram Representing a Fish Sample Analyzed for PFHS (Exygen ID: C0049478, Data Set: 061605E) .......... ....... .................................. 58 Figure 9. Typical Calibration Curve for PFOS in M ethanol.............................................59 Figure 10. Non-Extracted Standards o f PFOS in Methanol, 0.0005 pg/mL and 0.001 pg/mL, Respectively........................................... ...... ..................... .......... 60 Figure 11. PFOS in Control Fish Blank, 0.00625 pg/mL Fortified Control Fish Spk A, and 0.025 pg/mL Fortified Control Fish Spk B, Respectively................................61 Figure 12. Chromatogram Representing a Fish Sample Analyzed for PFOS (Exygen ED: C0049478, Data Set:061605E)..................................................... 62 Exygen Research Page 9 of 132 interim Report #10-Analysis of Fish and Clam Samples Exygen Study No.: P000113 LIST OF APPENDICES Page Appendix A Study Protocol P0001131 (Exygen Study No. P0001131) with Analytical Method V00017S3: "Method o f Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS", Protocol Amendments 1and 2, and Deviations 2 and 3 .............................. ............................................................................ ......... ...63 Exygen Research Page 10 of 132 Interim Report #10~Analy$is of Fish and Clam Samples Exygen Study No.: P001131 1.0 SUMMARY Exygen Research extracted and analyzed fish and clam samples for the determination of perfluorobutanesulfonate (PFBS), perfluorohexanesulfonate (PFHS), and perfluorooctanesulfonate (PFOS) according to Exygen Method V0001783 (Appendix A), The limit of quantitation (LOQ) for PFBS, PFHS, and PFOS in fish and clams was 0.2 ng/g. For one sample (C0048133) only one gram o f fish was used for extraction, so the limit o f quantitation for PFBS, PFHS, and PFOS in this fish sample was 1.0 ng/g. The initial analytical results associated with fifty-one fish samples are not reported (NR) in Table I because the endogenous levels o f PFOS were too high to calculate accurate matrix spike recoveries. These samples were re-extracted and the results of the reanalyses are summarized in Tables II and V. Analytical results and assessed accuracies for the analysis o f PFBS, PFHS and PFOS found in fish samples are summarized in Table I. Analytical results and assessed accuracies for the analysis o f PFOS found in the re-extracted fish samples are summarized in Table II. Analytical results and assessed accuracies for the analysis of PFBS, PFHS and PFOS found in clam samples are summarized in Table III. Laboratory matrix spikes that were in the appropriate concentration range (primary concentration less than three times the spiking level) were used to calculate the assessed accuracies. The average percent recoveries standard deviations for PFBS, PFHS, PFOS in fish samples were 79% 13%, 94% 12%, and 85% 20%, respectively. Fortification recoveries for PFBS, PFHS, and PFOS in fish samples are summarized in Table IV. The average percent recovery standard deviation for PFOS in the re-extracted fish samples was 109% 20%. Fortification recoveries for PFOS in the re-extracted fish samples are summarized in Table V. The average percent recoveries standard deviations for PFBS, PFHS, PFOS in clam samples were 84% 10%, 93% 13%, and 73% 16%, respectively. Fortification recoveries for PFBS, PFHS, and PFOS in clam samples are summarized in Table VI. The sample IDs assigned by the client follow this formula: Dxx-x01-xxxxxx-x-04xxxx. The first string begins with D for Decatur, Alabama and the xx defines the sampling area within the Wheeler Reservoir of the Tennessee River where: DL3 = Up river Letter o f Intent (LOI) sampling location LOC-3 DL2 = Cross river LOI sampling location LOC-2 DL1 = Downstream Fox Creek mouth LOI sampling location LOC-1 DBC = Bakers Creek mouth DOU - Cove at 3M WWTP outfall DMC = Downstream Mallard Creek mouth Exygen Research Page 11 of 132 Interim Report #1-Analysis of Fish and Clam Samples Exygen Study No.: P001131 The second string defines the general category o f the biota sample and the sampling round where: F01 = Fish sample first round 101 = Invertebrate sample first round The third string defines the species (first two characters), the sample type (third character), and the sample number (last three characters) within that sampling area and type of sample where: IP TMchannel catfish (Ictalurus punctatus) MS ~ largemouth bass (Micropterus salmoides) CF = Asiatic clam (Corbicula fluminea) F --Filet tissue sample W --Whole body sample 001 through 005 --sample number within the sampling area and sample type The fourth string (single character) additionally defines the sample type where: 0 = Primary field sample 1 = Field duplicate sample 2 - Equipment rinseate blank sample The fifth string defines the sample preparation date in YYMMDD format. Examples o f this sample ID system are: Sample DBC-F01-IPF004-0-041021 is the filet tissue sample prepared on October 21, 2004 from the fourth channel catfish collected at the Bakers Creek mouth sampling location. Sample DL1-F01-MSW002-0-041104 is the whole body sample prepared on November 4, 2004 from the second largemouth bass collected at the Fox Creek mouth sampling location. 2 0 O H.l F.f rpI yt p The objective of the analytical part o f this study was to determine levels of perfluorobutanesulfonate (PFBS), perfluorohexanesulfonate (PFHS), and perfluorooctanesulfonate (PFOS) in fish and clams according to Protocol P0001131 (Appendix A). Exygen Research Page 12 of 132 Interim Report #10-Analysis of Fish and Clam Samples Exygen Study No.: P0001131 3,0 INTRODUCTION This report details the results of the analysis for the determination of PFBS, PFHS, and PFOS in fish and clams using the analytical methods entitled, "V0001783: Method o f Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS." The study was initiated on November 05, 2004, when the study director signed protocol number P0001131. The analytical start date for this interim report was December 27, 2004, and the analytical termination date for this interim report was April 17, 2006. 4.0 ANALYTICAL TEST SAMPLES Forty fish samples (Exygen ID C0048126-C0048165) were received on dry ice on October 23, 2004 from Tim Frinak at Weston Solutions, Inc. Fifty-six fish samples (Exygen ID C0049450-C0049505) were received on dry ice on November 6, 2004 from Charles Young at Weston Solutions, Inc. Twenty-six fish samples (Exygen ID C0056354-C0056479) were received on dry ice on January 13, 2005 from Tim Frinak at Weston Solutions, Inc. Six clam samples, consisting o f whole clams from each sample site (Exygen ID C0056480-C0056485) were received on dry ice on January 13, 2005 from Tim Frinak at Weston Solutions, Inc. A whole Whiting (Family: Gadidae) was purchased from a local grocer by Ed Cams on December 20, 2004 and was used as the control fish (Exygen ID C0054391). The samples were logged in by Exygen personnel and placed in frozen storage. Sample login and chain o f 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 standards, PFBS and PFHS, were supplied by 3M. PFBS was received from 3M at Exygen on July 6, 2000, and May, 13, 2005. PFHS was received from 3M at Exygen on January, 20, 2003. PFOS was purchased from Fluka Corporation and was received at Exygen on April, 23, 2003. The available information for the reference materials is listed below. PFBS and PFHS were stored frozen and PFOS was stored refrigerated. Exygen Research Page 13 of 132 Interim Report #10-Analysis of Fish and Clam Samples Exygen Study No.: P0001131 Compound PFHS PFBS PFHS PFOS Exveen Inventory No. SP0000252 SP0005726 SP02401 SP02694 Lot # 101 101 SE036 430180-1 Puritv (%) 96.7 96.7 98.6 101.2 Expiration E 12/04/06 12/04/06 10/18/06 10/31/07 The molecular structures o f PFBS, PFHS and PFOS are given below: PFBS Chemical Name: Perfluorobutanesulfonate Molecular Weight: 338 supplied as the potassium salt (C4F9SO3IC ) Transitions Monitored: 299 - 99 Structure: PFHS Chemical Name: Perfluorohexanesulfonate Molecular Weight: 438 supplied as the potassium salt (CFu SCVK*) Transitions Monitored: 399 80 Structure: FFF FFF F SO 3 FFF FFF PFOS Chemical Name: Perfluorooctanesulfonate Molecular Weight: 538 supplied as the potassium salt (CgFiySOs'K4) Transitions Monitored: 499 80 Structure: FFFF FFFF F SO3 FFFF Hxygen Research Page 14 of 132 interim Report #10-Analysis of Fish and Clam Samples Exygen Study No. : F0001131 6.0 DESCRIPTION OF ANALYTICAL METHOD The analytical method "V0001783: Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS" was used for the fish and clam samples in this study. 6.1 Extraction Procedure 6.1.1 Sample Preparation Before the samples could be weighed for the extraction, they had to be processed. To process, the frozen samples were placed into a food processor and homogenized with dry ice. The samples were transferred to one-gallon Ziploc bags and placed in frozen storage with bag left open to allow the dry ice to sublime. After sublimation, the sample bags were sealed and remained in frozen storage until time o f analysis. Sample processing records are located in the Sample Information section o f the raw data package. 6.1.2 Glassware Preparation The 125 mL pear-shaped flasks were silanized by first rinsing the flask with 30% dimethyldichlorosilane in toluene solution. The flasks were rinsed once with toluene followed by three rinses with methanol. The flasks were allowed to air dry before use. 6.1.3 Column Preparation The 20 mL columns were packed at Exygen in sequence with 2 grams florisil, 2 grams silica gel, 2 grams o f carbon, and 1 gram LC-NFL. The columns were conditioned on the day o f extraction with 20 mL of methanol followed by 20 mL o f acetonitrile. All washes were discarded. These clean-up columns were used to remove matrix interference and not to retain PFBS, PFHS, and PFOS. Additional details about the column packing materials can be found in the raw data package associated with this report. 6.1.4 Sample Extraction A 5-gram* portion o f fish or clam sample was weighed into a fiftymilliliter centrifuge tube for the extraction. After fortification of appropriate samples, 30 mL o f acetonitrile was added to the samples. The samples were allowed to shake on a wrist action shaker for -1 5 minutes. The samples were placed into a freezer for -1 hour. Freezing o f the Exygen Research Page 15 of 132 Interim Report #10-Analysis of Fish and Clam Samples Exygen Study No.: P01131 samples allowed the fats and proteins to precipitate. The samples were centrifuged for -10 minutes at -2000 rpm. The supernatant was then loaded onto the conditioned clean-up column fitted inside the silanized pear shaped flask. The eluate was collected in the silanized pear-shaped flask. This process removed the fats and proteins from the extract. Ten milliliters of acetonitrile was added to the samples left in the centrifuge tube. The remaining sample in the tube was homogenized using a tissumizer for -30 seconds. The tissumizer was rinsed with ten milliliters of acetonitrile. The rinse was added to the sample tube. The samples were allowed to shake on a wrist action shaker for another 10 minutes. The samples were placed into a freezer for -1 hour. The samples were centrifuged again for -10 minutes at -2000 rpm. The supernatant was then loaded onto the same clean-up column. The eluate was collected into the same pear-shaped flask. The column was washed with 20 mL o f acetonitrile, collecting the wash in the pear-shaped flask. Approximately 3 or 4 drops of l-octanol was added to the extracts in the flasks. The samples were evaporated using a rotary evaporator at reduced pressure. The l-octanol held the analytes in the pear-shaped flask while the solvent evaporated. Two milliliters o f 2% ascorbic acid in methanol was added to the flasks to make final volume. The flask was swirled to dissolve and mix the sample. The sample was transferred to a HPLC vial using a disposable pipet. Each sample was analyzed by LC/MS/MS electrospray. *Due to lack o f sample, a one-gram portion o f sample C0048133 (DMC-F01-MSW010-041216) was used. All other samples were weighed out in 5-gram portions. 6.2 Preparation of Standards and Fortification Solutions Individual stock standard solutions o f PFBS, PFHS and PFOS were prepared as specified in Exygen method V0001783. The stock standard solutions were prepared at a concentration of 100 pg/mL by dissolving 10 mg o f each o f the standards (corrected for purity and salt content) in methanol. From these solutions, mixed 1.0 pg/mL fortification standard solutions were prepared by taking 1 mL o f each o f the appropriate stock solutions and bringing the volume up to 100 mL with methanol. By taking 10 mL o f the mixed 1.0 pg/mL fortification standard and bringing the volume up to 100 mL with methanol, a mixed 0.1 pg/mL fortification standard was prepared. A mixed stock standard solution of PFBS, PFHS, and PFOS was also prepared. The stock standard solution was prepared at a concentration o f 1000 pg/mL by dissolving 100 mg o f each o f the standards (corrected for purity and salt content, if necessary) in methanol. From this solution, a 100 pg/mL fortification standard solution was prepared by taking 10 mL of the stock and bringing the volume up to 100 mL with methanol. By taking 10 mL o f the 100 pg/mL fortification standard and bringing the volume up to 100 mL with methanol, a 10 pg/mL fortification standard was prepared. By taking 10 mL o f the 10 pg/niL fortification standard and bringing the volume up to 100 mL with Exygen Research Page 16 of 132 Interim Report #10-Anaysis of Fish and Clam Samples Exygen Study No.: P000113I methanol, a 1.0 pg/mL fortification standard were prepared. By taking 10 mL o f the 1.0 pg/mL fortification standard and bringing the volume up to 100 mL with methanol, a 0.1 pg/mL fortification standard was prepared. A set of non-extracted standards containing PFBS, PFHS and PFOS was prepared in methanol. The following concentrations were prepared: Cone, of Fort Fort Solution Volume (Fg/mL)1 (mL) 1.0 5.0 1.0 2.5 1.0 1.0 0.05 10 0.025 10 0.01 10 0.005 10 1of PFBS, PFHS and PFOS Volume of Fortified Sample (mL) 100 100 100 100 100 100 100 Final Cone, of Calibration Std. (pg/mL) 0.05 0.025 0.01 0.005 0.0025 0.001 0.0005 The stock standard solution and all fortification and calibration standard solutions were stored in a refrigerator (4 2C) when not in use. Documentation o f standard preparation is located in the raw data package associated with this interim report. 6.3 Chromatography Quantification o f PFBS, PFHS and PFOS was accomplished by LC/MS/MS electrospray. The retention times o f PFBS, PFHS and PFOS were ~ 4.7 mins, - 10.8 mins, and - 12.9 mins, respectively. 6.4 Instrument Sensitivity The smallest standard amount injected during the chromatographic run had a concentration of 0.0005 pg/mL of PFBS, PFHS and PFOS. 6.5 Description of LC/MS/MS Instrument and Operating Conditions Instrument: API 4000 Biomolecular Mass Analyzer Interface: Turbo Ion Spray Liquid Introduction Interface Computer: DELL OptiPlex GX400 Software: Windows NT, Analyst 1.4.1 HPLC: Hewlett Packard (HP) Series 1100 HP Quat Pump HP Vacuum Degasser HP Auto sampler Exygen Research Page 17 o f ! 32 Interim Report #10-Analysis of Fish and Clam Samples Exygen Study No.: P0G0131 HP Column Oven HPLC Column: Thermo Fluophase RP, 50 mm x 2.1mm Column Temp.: -30 C Injection VoL: 15 pL Mobile Phase (A): 2 mM Ammonium Acetate in water Mobile Phase (B): Methanol Time (min! 0.0 TO 8.0 10.0 11.0 18.0 Total run time: -18 min Flow Rate: 0.3 mL/min %A 65 65 25 25 65 65 %B 35 35 75 75 35 35 Ions monitored: Analyte PFBS PFHS PFOS Mode negative negative negative Transition Monitored 299 99 399 80 499 80 Approximate Retention Time (min) -4.7 min. - 10.8 min. -12.9 min. 6.6 Quantitation and Example Calculation 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 1/x fit weighted linear regression) by Analyst software using six concentrations o f standards. The concentration was determined from the following equations. Equation 1 calculated the amount of analyte found (in ng/mL, based on peak area) using the standard curve (linear regression parameters) generated by the Analyst software program. Equation 1: Analyte found (ng/mL, wet weight) - (Peak area - intercept) x DF slope Where: DF = Dilution Factor, factor by which the final volume was diluted, if necessary. Exygen Research Page 18 of 132 Interim Report #10-Analysis of Fish and Clam Samples Exygen Study No.: P0001131 Equation 2 was used to convert the amount o f PFBS, PFHS and PFOS found in ng/mL to ng/g (ppb). Equation 2: Analyte found (ppb, wet weight) = [Analyte found (ng/mL) x Final volume (2 mL)] sample weight (5 g) For samples fortified with known amounts o f PFBS, PFHS and PFOS prior to extraction, Equation 3 was used to calculate the percent recovery. Equation 3: Recovery (%) = (analyte found (ng/mL) - analyte in control (ng/mL)) x!00% amount added (ng/mL) Note: For the analyte recovery calculation, the "control" is the unspiked aliquot o f the primary field sample. An example of a calculation using an actual sample follows (calculation is for PFBS only): Fish sample Exygen ID: C0049478 Spk E (Set: 061605E), fortified at 10 ng/g with where: peak area = 280750 intercept = 3070 slope = 13100 dilution factor -1 ng/g PFBS added (fort level) = 10 amt in corresponding sample - NQ (not quantifiable) final volume (mL) =2 sample weight (g) =5 From equation 1: Analyte found (ng/mL, wet weight) = [280750 - 30701 x 1 13100 = 21.2 ng/mL From equation 2: Analyte found ng/g (ppb) - (21.2 ng/mL x 2 mL) 5g = 8.48 ng/g (ppb) Exygen Research Page 19 of 132 Interim Report #10~Analysis of Fish and Clam Samples Exygen Study No.: P0001131 From equation 3: % Recovery - 18.48 ng/g - 0 ng/g) x 100% 10ng/g = 85 % 7.0 EXPERIMENTAL DESIGN For fish and clam samples designated as laboratory matrix spikes, PFBS, PFHS, and PFOS were added at a known concentration to the samples in the laboratory after the samples were weighed for extraction. The fish samples were extracted in thirty sets, five that were re-extraction sets. The clam samples were extracted in one set. Each fish or clam set included one control fish matrix blank and two control fish matrix blanks fortified at known concentrations. Two sets of fish contained three samples, one set o f fish contained four samples, one set o f fish contained seven samples, and twenty-one sets o f fish contained five samples. Three sets o f re-extracted fish contained ten samples each. One set o f re-extracted fish contained twelve samples. One set of re-extracted fish contained nine samples. One set o f clams contained six samples. Accuracies were assessed for each sample by reviewing the individual QC results obtained for each sample. 8.0 RESULTS Analytical results and assessed accuracies for the analysis of PFBS, PFHS and PFOS found in fish samples are summarized in Table I. Please note that the chromatography for PFBS was poor for this matrix. Exygen recognizes this and great care has been taken to integrate each data set consistently. Analytical results and assessed accuracies for the analysis of PFOS found in the re-extracted fish samples are summarized in Table II. Analytical results and assessed accuracies for the analysis of PFBS, PFHS and PFOS found in clam samples are summarized in Table III. The average percent recoveries standard deviations for PFBS, PFHS, PFOS in fish samples were 79% 13%, 94% 12%, and 85% 20%, respectively. Fortification recoveries for PFBS, PFHS, and PFOS in fish samples are summarized in Table IV. The average percent recovery standard deviation for PFOS in the re-extracted fish samples was 109% 20%. Fortification recoveries for PFOS in the re-extracted fish samples are summarized in Table V. The average percent recoveries standard deviations for PFBS, PFHS, PFOS in clam samples were 84% 10%, 93% 13%, and 73% 16%, respectively. Fortification recoveries for PFBS, PFHS, and PFOS in clam samples are summarized in Table VI. Exygen Research Page 20 of 132 Interim Report #10-Analysis of Fish and Clam Samples Exygen Study No.: P0001131 9.0 CONCLUSIONS The fish and clam samples were successfully extracted and analyzed for PFBS, PFHS and PFOS according to analytical method V0001783. 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 o f all raw data, as well as a signed copy o f the final analytical report and all original facility-specific raw data, will be retained in the Exygen Research archives for the period o f time specified in EPA TSCA Good Laboratory Practice Standards 40 CFR 792. Exygen Research Page 21 of 132 Interim Report #10~Analysis of Fish and Clam Samples Exygen Study No.: PO1131 TABLES Exygen Research Page 22 of 132 Interim Report #10-Analysis of Fish and Clam Samples Exygen Study No.: P0001131 Table I. Summary ofPFBS, PFHS and PFOS in Fish Samples Exygen ID C lie n t S am ple ID C4 Sulfonate PFBS Perfiuorotiutan esu ifonata Analyte Assessed Found Accuracy (ng/g) wet wi. (+/- %) C6 Sulfonate PFHS Parfluoro hexanesuifonate A n a ly te Found Assessed Accuracy (ng/g) wet wt. {+/- %} C8 Sulfonate PFOS Perfluorocictanesutfonats A n a ly te Found Assessed Accuracy (ng/g) wet wt. {+/-% } C0048126 DL3-F1-IPFG01-0-041021 0.309 40 ND 30 C0048126 Rep D L3-F 1-IP F 0 0 1-0-041021 * 0.316 40 ND 30 CG048127 D L 3 -F 0 1-IPFOG2-0-041021 0.265 40 ND 30 C0048127 Rep D L3-F 1-IPFO2--041021 * 0.382 40 ND 30 C0048128 D M C -FQ 1-SP F 0 0 1-0-041021 0.330 50 ND 50 C0048128 Rep DM C-F01-IPF001-0-041021* 0.366 50 ND 50 C0048129 D M C -F 1-SP F 0 0 2 -0 -0 4 1021 0,712 30 ND 30 C0048129 Rep D M C -F 01-IPF2--041021 * 0.792 30 ND 30 00048130 C0048130 Rep DMC-FQ 1-IPW 01-0-041021 D M C -F 0 1 -IP W 01-0-041021 * 1.08 1.06 40 ND 30 40 0.297 30 C0048131 DM C -F 0 1-1P W 00 2-0-041021 0.968 50 ND 30 C0048131 Rep DMC-FQ 1-I P W 0 0 2 -0 -0 4 1021 * 1.34 50 ND 30 00048132 DMC-F01-MSW 001-0-041021 0.218 30 0.524 30 C 0 0 4 8 1 3 2 R ep D M C -F 0 1-M S W 0 0 1-0-04 102 1* 0.202 30 0.484 30 C0048133A DM C -F01-M SW 002-0-041021 C 0 0 4 8 1 3 3 R e p A D M C -F 0 1-M S W 02 -0-0410 2 1* NDA NDA 30 30 NDA NDA 30 30 C0048134 DL2-F01 -IP F001-0-041021 0.660 50 ND 40 C048134 Rep DL2-F01-IPF001 -0-041021 * 1.00 50 ND 40 C4813S D L 2 -F 0 1-SPF02-0-041021 0.816 50 ND 40 C0048135 Rep D L 2 -F 0 1-IP F 0 0 2 -0 -0 4 1021 * 0.952 50 ND 40 C0048136 D L 2 -F 0 1-IP F 0 0 3 -0 -0 4 1021 0.247 30 ND 30 C048136 Rep D L 2 -F 0 1-IP F 003-0- 41021 * 0.216 30 ND 30 C0048137 D L2-F01-IP F004-0-041021 0.346 40 ND 30 C048137 Rep DL2-F1-IP F004-0-041021* 0.330 40 ND 30 C0048138 D L 2 -F 0 1 -IP F 0 5-0 -041021 0.338 40 ND 30 C0048138 Rep DL2-F01-SPF005--Q41021 * 0.318 40 ND 30 00048139 DL2-F01-IPW 01-0-041021 0.326 30 ND 30 C048139 Rep D L 2 -F 0 1-1P W 0 0 1-0 -0 4 10 2 1* 0.400 30 ND 30 C0048140 D L 2 -F 0 1-M SF001 -0-041021 0.333 30 ND 30 C0048140 Rep DL2-F1-M SF01-0 -0 4 10 2 1* 0.348 30 ND 30 C048141 DL2-F1-M S F 002-0-0410 2 1 0.408 40 ND 30 C0048141 Rep D L 2 -F 0 1-M SF 02-0- 41021* 0.398 40 ND 30 C048142 D L 2 -F 0 1-M S F 0 0 3 -0 -0 4 1021 0,416 40 ND 40 C48142 Rep D L 2-F 01-M S F 003 -0-0 410 2 1* 0.376 40 ND 40 C48143 D L2-F01-M S F004-0-041021 0.345 30 ND 30 C0048143 Rep D L2-F01-M S F004-0-0410 2 1* 0.349 30 ND 30 5.16 5.64 4.16 4.56 7.40 6.36 14.1 17.0 70,0 56.4 34,4 34.2 270 280 530 548 3.45 2.86 4.72 5,04 2.01 2.08 4.52 4,12 2.46 2.80 5.88 6.12 77,2 67.6 94.8 98.4 41.6 40.0 83.6 83.2 30 30 40 40 50 50 50 50 40 40 30 30 30 30 30 30 50 50 50 50 30 30 40 40 30 30 30 30 30 30 40 40 40 40 30 30 'Laboratory Duplicate A 1,0 g of sample used for extraction instead of 5.0 g. ND Not detected at or above Limit of Quantitation which is 0.2 ng/g {wet weight}. NDA = Not detected at or above Limit of Quantitation which is 1.0 ng/g {wet weight). NR = Not reported due to quality control result failures. Exygen Research Page 23 of 132 Interim Report #10~Anaysis of Fish and Clam Samples Exygen Study No.: P001131 Table I. Summary of PFBS, PFHS and PFOS in Fish Samples (continued) Exygen ID 00048144 C0048144 Rep C0048145 C48145 Rep C048146 C0048146 Rep C0048147 C48147 Rep C0048148 C0048148 Rep C0048149 C048149 Rep C0048150 C0048150 Rep C0048151 C0G48151 Rep C048152 C048152 Rep C0048153 C048153 Rep 00048154 C0048154 Rep 00048155 C0048155 Rep 00048156 00048156 Rep C0048157 C0048157 Rep C0048158 00048158 Rep C048159 00048159 Rep C0048160 C0048160 Rep C048161 C048161 Rep C0048162 C0048162 Rep Client Sample SD C4 Sulfonate PFBS Psffltiorobutanesutfonate Analyte Assessed Found Accuracy (rq/g} wet wt. (+/- %} 6 Sulfonate PFHS PerfluofQhaxan&su Ifonate Analyte Assessed Found Accuracy {ng/g}wet wt. .....<+'-%)... - C8 Sulfonate PFOS Peffiuofooctanesulfonate Analyte Assessed Found Accuracy (ng/g)wet wt. <+/-%} DL1-F01 -IPF001 -0-041021 DL1-F01-IPF01 -0-041021* 0.432 0.428 30 0.261 30 0,252 30 30 33.4 32.6 40 40 DL3-F01 -IPF03--41021 0.448 30 ND 30 DL3-F01-iPF003-G-041021* 0.440 30 0.229 30 3.55 3.35 30 30 DL3-F1-PFG04-0-041021 0.784 40 ND 30 0.382 30 DL3-F01-1PF04--Q41021* 0.900 40 ND 30 0.230 30 DL3-F1-i PF05-0-041021 1.53 60 ND 30 5.48 40 DL3-FQ1-IPF005-0-041021 * 1.07 60 ND 30 5.64 40 DBC-F01-IPF01-0-041021 DBC-F01-IPF001-0-041021 * 1,34 1.36 50 0.692 50 0.680 30 30 NR NR NR NR D8C-F1-I PF002-0-041021 1.43 50 ND 30 110 30 DBC-F01-IPF002-0-041021* 0,756 50 ND 30 109 30 D8C-F01-1PF003-0-041021 0.996 50 0.644 30 DBC-F01-1PF003--041021 * 0.952 50 0.624 30 NR NR NR NR DBC-F01-IPW001 -0-041021 0.864 40 2.21 30 NR NR DBC-F01-1PW01-0-041021* 0.508 40 2.20 30 NR NR DBC-F01-1PW002-0-041021 0.700 40 9.12 30 NR NR DBC-F01-1PW002-0-041021 * 0.736 40 8.96 30 NR NR DBC-F01- IPW003-0-041021 DBC-F01-IPW003-0-041021 * 0.736 0.760 30 30 6.76 7.44 30 30 NR NR NR NR DOU-F01-1PF001-0-041021 DOU-F01-1PF001 -0-041021 * 4.36 4.24 40 1.02 40 0.972 30 30 NR NR NR NR DO-F01 -fPF002-0-41021 6.76 40 0.792 30 NR NR DOU-F01-IPF002-0-041021 * 6.28 40 0.720 30 NR NR DOU-F01- 1PF003-0-041021 5,88 40 1.02 30 NR NR DQ-F01-IPF003-0-041021* 5.96 40 1.02 30 NR NR DOU-F01-IPF004-0-041021 6.12 30 2.30 30 DOU-F01-IPF04-0-041021* 6.28 30 2.38 30 NR NR NR NR DOU-F01-IPF005-Q-041021 OOU-F01-PF005-0-041021 * 4,56 4.36 30 0.956 30 30 0.892 30 NR NR NR NR DOU-F01-IPW 0 0 1-0-041021 DOU-F01- IPW001-0-041021* 2.89 2.81 30 0.948 30 30 0.964 30 NR NR NR NR DOU-F01-IPW002-0-041021 18.4 50 3.69 30 NR NR DOU-F01-IPW002-0-41021 * 19.3 50 3.70 30 NR NR DL2-F1-MSW01--41021 0.203 30 ND 30 120 50 DL2-F01-MSWQ01-0-041021 * ND 30 ND 30 120 50 DL2-F1-MSW002-0-041021 ND 30 ND 30 102 30 DL2-F01-M5WO02-O-G41021 * NO 30 ND 30 100 30 ` Laboratory Duplicate ND = Not deiected at or above Limit of Quantitation which is 0.2 ng/g (wet weight). NR = Not reported due to quality control result failures. Exygen Research Page 24 of 132 Interim Report #10~Analysis of Fish and Clam Samples Exygen Study No.: P00U31 Table L Summary of PFBS?PFHS and PFOS in Fish Samples (continued) Exygen ID Ciient Sample ID C4 Sulfonate PFBS Perfluorcbutanesuifonaie Analyte Assessed Found Accuracy (ng/g) wet wt. (+/- %> 0 6 Sulfonate PFHS Perfluorohexartesu Ifonata Analyte Found Assessed Accuracy (ng/gj wet wt. {+/- %) C8 Sulfonate PFOS Perfluorooctanesu Sfonato Analyte Found Assessed Accuracy (ng/g) wet wt. (+/- %) C0048163 DL2-F01-MSW03-Q-041021 ND 30 ND 30 C0048163 Rep DL2-F01-MSWG3-0-41021* ND 30 ND 30 C0048164 DL2-F01-MSW004-0-041021 ND 30 ND 30 C0048164 Rep DL2-F01-MSW004-0-041021 * ND 30 ND 30 C0048165 DL2-F1-MSW005-0-041021 ND 30 ND 30 C0048165 Rep DL2-FG1-MSW05-0-041021 * ND 30 ND 30 CQ4945 C0049450 Rep DBC-F1-MSW001-0-041104 DBC-F01-MSW01-0-041104* 0,888 0.852 30 30 8.32 8.32 30 30 CQ49451 C0049451 Rep DBC-FO1-MSWOO2-Q-0411O4 DBC-F01-MSW02-0-Q4114* 0.868 0,864 30 30 3,43 3.35 30 30 C0049452 C49452 Rep DBC-F01-MSW003-0-041104 DBC-F01-MSW0Q3-0-041104' 0.860 0.784 30 30 3.62 3.64 30 30 C0049453 C0049453 Rep DBC-F01-MSW004-0-41104 DBC-F01-MSW004-0-41104* 0.884 0.880 30 30 9,72 9,80 30 30 C049454 C0049454 Rep DBC-F01-MSW005-0-041104 DBC-F01-MSW005-0-041104* 0.832 0.832 30 30 2.77 2,66 30 30 C049455 C0049455 Rep DBC-F01-MS F001-0-041104 DBC-F1 -MSF001-0-041104* 0.820 0.788 30 30 3.58 3.46 30 30 C0049456 DBC-F01-MSF002-0-041104 0,768 30 1.94 30 C49456 Rep DBC-FG1-MSF002-0-41104* 0.760 30 1.90 30 C0049457 DBC-F1-IPF4-G-041104 0.504 30 0.584 30 C0049457 Rep DBC-F01-1PF04-0-041104* 0.520 30 0.540 30 C04458 DBC-F01-PF005-0-041104 1.60 30 2.15 30 C049458 Rep DBC-P01-iPF005-0-041104* 1.64 30 2.18 30 C0049459 C0049459 Rep DBC-F01-IPW004-0-041104 DBC-F01-IPW004--041104* 0.484 0.444 30 30 3.28 3,28 30 30 C0049460 C0049460 Rep DBC-F01-IPW005-0-041104 DBC-F01-IPW005-0-041104* 0.572 0.532 30 30 2.10 2.10 30 30 C049461 DOU-F01-1PW03-0-041104 3.12 30 6.00 30 C0049461 Rep DOU-F1-1PW003-0-041104* 3.05 30 5.92 30 C049482 DOU-F01-I PW004-0-041104 3.49 30 7.12 30 COO49462 Rep DOU-F01-tPW004-0-041104* 3.11 30 6.84 30 C0049463 DOU-F1-IPW005-0-041104 2.97 30 5.12 30 C0049463 Rep DOU-F01-tPW005-0-041104* 2.90 30 5.32 30 C0049464 DL2-F01-1PW2-0-041104 1.04 30 ND 30 C049464 Rep DL2-F01-i PW002-0-041104* 0.980 30 ND 30 C0049465 C049465 Rep D L 1-F 0 1-MSW 0 0 1-0-041104 DL1-F01-MSW 001-0-041104* 1.20 1.24 30 0.896 30 0,932 30 30 92.4 91,6 88.0 94.8 168 174 NR NR NR NR NR NR NR NR NR NR NR NR NR NR 142 152 NR NR NR NR NR NR NR NR NR NR NR NR 8,20 6.60 NR NR 30 30 30 30 30 30 NR NR NR NR NR NR NR NR NR NR NR NR NR NR 30 30 NR NR NR NR NR NR NR NR NR NR NR NR 40 40 NR NR ` laboratory Duplicate ND = Not detected at or above Limit of Quantitation which is 0.2 ng/g (wet weight}. NR = Not reported due to quality control result failures. Exygen Research Page 25 of 132 Interim Report #10-Analysis of Fish and Clam Samples Exygen Study No.: P001131 Table I. Summary of PFBS, PFHS and PFOS in Fish Samples (continued) Exygen ID 00049466 C0049466 Rep C0049467 C049467 Rep 00049468 C49468 Rep C0049469 C049469 Rep 00049470 C0049470 Rep C0049471 C0049471 Rep C0049472 C049472 Rep C049473 C0049473 Rep 00049474 C0049474 Rep C0049475 00049475 Rep C0G49476 C49476 Rep C049477 C0049477 Rep C049478 C0049478 Rep C049479 00049479 Rep 00049480 C49480 Rep 00049481 C049481 Rep 00049482 00049482 Rep 00049483 C0049483 Rep C0049484 C0049484 Rep Client Sample ID C4 Sulfonate PFBS Perfitiorobtitan esutfonate Analyte Assessed Found Accuracy (ng/g)wet wt. {+/- %} CS Sulfonate PFHS Perfluorohexanesutfenate Analyte Assessed Found Accuracy (ng/g)wet wt. (*/- %) C8 Sulfonate PFOS Perfiuorooctanesuifonat# Analyte Assessed Found Accuracy (ng/g) wet wt. {+/-%> DL1-F01 -MSWO02-Q-O41104 1.30 30 1.72 30 DL1-FQ1-MSWQ2-0-041104* 1.15 30 1.54 30 DL1-F01 -MSW003-0-041104 0,560 30 1.13 30 DL1-FQ1-MSW003-0-041104* 0.528 30 1.14 30 D L 1-F1-MS W04-0-041104 DL1-FQ1-MSW0Q4-0-041104* 1,22 1.12 30 0.908 30 30 0.880 30 D L 1-FQ1-M SW 005-0 -041104 1.22 30 1.18 30 DL1-F1-MSW5-G-041104* 1.20 30 1.18 30 DL1-F01-MS F001-0-041104 DL1-F1 -M SF001-0-041104* ND NO 30 0.315 30 30 0.299 30 L1-F1-MSF002-0-41104 DL 1-F01-MSF002-0-041104* ND ND 30 0.624 30 30 0.604 30 DL1-F01-MSF003-0-41104 0.203 30 0.652 30 DL 1-F01-MSF0G3-G-Q41104* ND 30 0.676 30 DL1-F01-MSF004-0-041104 DL1-F01-MSF004-0-041104* ND ND 30 0.532 30 30 0.536 30 DL1-F01-ySF005-0-041104 DL 1-F01-MSF005-0-041104* ND ND 30 0.684 30 30 0.644 30 DL1-F01-1PW001-0-041104 0.772 30 0.556 30 DL 1-F01-IPW001-0-041104* 0.870 30 0,744 30 DL1-F01-1PW002-0-0411Q4 0.468 30 0,536 30 DL1-F01-IPW002-0-041104* 0.472 30 0,516 30 DL1-F01-IPW003-0-041104 DL 1-F01-IPW003-0-041104* ND ND 30 0,209 30 30 0.247 30 DL1-F01-PW004-Q-041104 DL1-F014PW0O4--41104* ND ND 30 0,238 30 30 0.246 30 DL1-F1-IPF02--041104 DL 1-F01-IPF002-0-041104* ND ND 30 0.222 30 30 0.247 30 DL-1 ~FG1-IPF3-0-041104 DL-1-F01-IPF003-0-041104* ND ND 30 0,265 30 30 0.262 30 DL-1-F01-IPF0Q4-0-41104 ND 30 ND 30 DL-1-F01-IPF04-0-41104* ND 30 ND 30 DL-1 -F01-IPF005-0-041104 DL-1-F01-IPF005-0-041104* ND ND 30 0.373 30 30 0,386 30 DL3-F01-MSW 0 0 1-0-041105 ND 30 ND 30 DL3-F01-MSW001-0-041105* ND 30 ND 30 DL3-F01 -MSW002-0-041105 ND 30 ND 30 DL3-F01-MSW002-0-041105* ND 30 ND 30 NR NR NR NR 181 186 NR NR 293 307 NR NR NR NR NR NR NR NR 17.4 18.9 15.7 15.6 9.36 9.52 59.6 57.2 11.1 11.2 20.9 21.4 7,60 7.36 8.72 8.56 139 137 160 162 NR NR NR NR 30 30 NR NR 30 30 NR NR NR NR NR NR NR NR 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 'Laboratory Duplicate NO = Not detected at or above Limit of Quantitation which is 0,2 ng/g (wet weight}, NR = Not reported due to quality control result failures. Exygen Research Page 26 of 132 Interim Report #10-Analysis of Fish and Clam Samples Exygen Study No.: P001131 Table I. Summary of PFBS, PFHS and PFOS in Fish Samples (continued) Exygen ID Client Sample ID C4 Sulfonate PFBS Perfluorobutanesuifonate Analyte Assessed Found Accuracy (ng/g)wet wt. (+/- %) C6 Sulfonate PFHS Rerfiuorohexa nesolfcnats Analyte Assessed Found Accuracy (ng/g)wet wt. (+/- %) 08 Sulfonate PFOS Ferfluorooctanesu ifcmate Analyte Assessed Found Accuracy (ng/g) wet wt. {+/- %) 00049485 DL3-F01-MSWG03-0-041105 ND 30 ND 30 C0049485 Rep DL3-F01-MSW03-G-041105* ND 30 ND 30 C0049486 C049486 Rep DL3-FQ1-MSW004-G-Q41105 DL3*F01 -MSW04-0-041105* ND ND 30 0.285 30 30 0.282 30 C0049487 C0049487 Rep DL3-F01-MSW005-0-041105 DL3-FG1-MSW5-0-G41105* ND ND 30 0.366 30 30 0,341 30 C0049488 DL3-F01-MSF001 -0-041105 ND 30 ND 30 C0049488 Rep DL3-F01-MSF0Q1-0-041105* ND 30 ND 30 C049489 DL3-F01-MSFQQ2-0-041105 ND 30 ND 30 C049489 Rep 0L3-F1-MSF2--041105* ND 30 ND 30 C04949 DL3-F01-MSF003--041105 ND 30 ND 30 C049490 Rep DL3-FQ1-MSFO03-0-O4115* ND 30 ND 30 C0049491 DL3-F01-MSF04-Q-Q41105 ND 30 ND 30 C0049491 Rep DL3-F1-MSF04-0-041105* ND 30 ND 30 C0049492 DL3-F1-MSF05--41105 ND 30 ND 30 C0049492 Rep DL3-F01-MSF005--041105* ND 30 ND 30 C0049493 CQ49493 Rep DMC-F01-IPW00 3-0-041105 DMC-F01-IPW003-0-041105* ND ND 30 0.386 30 30 0.334 30 C49494 DMC-F01-IPW004-0-041105 0.300 30 0.396 30 C0049494 Rep DMC-F01-IPW004-0-041105* 0,345 30 0.464 30 C049495 C0049495 Rep DMC-F1-IPW005-0-041105 DMC-F01-SPW005-Q-041105* ND ND 30 0.270 30 30 0,255 30 C0049496 DMC-F01 -MSW003-0-041105 C0049496 Rep DMC-F01-MSW003-0-041105* ND ND 30 0,592 30 30 0.560 30 C049497 DMC-F01-ySW004-0-041105 0.318 30 1.90 30 C0049497 Rep DMC-FQ1-MSW004-0-041105* 0.233 30 1,88 30 C049498 DMC-F01-MSW005--41105 C0049498 Rep DMC-F01-MSWQ05-0-41105* ND ND 30 0.980 30 30 0.936 30 C049499 C0049499 Rep DMC-F01-MSF002-0-041105 OMC-F01-MSF02-G-041105* ND ND 30 ND 30 30 0.367 30 C0049500 DMC-F01-MSF03-0-41105 ND 30 ND 30 C04950 Rep DMC-F01-MSF03-0-041105* ND 30 ND 30 C04951 C0049501 Rep DMC-FG1-MSF04-0-41105 DMC-F1-MSF004-0-41105* ND ND 30 0.254 30 30 0.228 30 CG04952 C004952 Rep DMC-F01-MSF005-0-041105 OMC- F1-MSF005-0-41105* ND ND 30 0.420 30 30 0.448 30 C004953 Df4C-F1-IPF003-0-041105 ND 30 ND 30 C0049503 Rep DMC-F01-SPF003-0-041105* ND 30 ND 30 146 144 198 199 173 168 116 115 116 111 75,6 72.0 130 134 81.6 83.6 61.2 63.6 68.8 69.6 64,4 62.8 354 330 259 269 NR NR NR NR 56.8 56.0 116 119 146 147 7.92 7.92 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 NR NR NR NR 40 40 30 30 30 30 30 30 ` Laboratory Duplicate NO = Not detected at or above Limit of Quantitation which is 0.2 ng/g (wet weight). NR =s Not reported due to quality control result failures. Exygen Research Page 27 of 132 Interim Report #10-Analysis of Fish and Clam Samples Exygen Study No.: P000113 Table I. Summary of PFBS, PFHS and PFOS in Fish Samples (continued) Exygen iD Client Sample ID 04 Sulfonate PFBS Perfluorobutanesu tfonaie Analyte Assessed Found Accuracy (ng/g)wet wt. (+/- %} 0 6 Sulfonate PFHS Perfiuoro hexanasu ifonate Analyte Assessed Found Accuracy (ng/g) wet wt. (+/- %) C8 Sulfonate PFOS Peffluorooctanasuifonate Analyte Assessed Found Accuracy (ng/g)wet wt. (*-% ) CG49504 DMC-F01-IPF04--41105 ND 30 ND 30 44.0 30 C0049504 Rep DMC-F1-IPF004-0-041105* ND 30 ND 30 45.6 30 00049505 DMC-F01-I PF05-0-Q41105 ND 30 ND 30 46.4 30 C04955 Rep DMC-F01-IPF05--41105* ND 30 ND 30 46.0 30 C0056354 DL2-F01-MSF005-0-41216 ND 30 ND 30 49,2 30 C0056354 Rep DL2-F01-MSF005-0-041216* ND 30 ND 30 51.6 30 C056455 DL2-F01 -IRW005-0-041215 ND 30 ND 30 13.2 30 C0056455 Rep DL2-F01-IPW005-0-041215* ND 30 ND 30 13.3 30 C0056466 DOU-F01-MSW002-0-041216 0.206 40 1.72 30 NR NR C0056456 Rep DOU-FQ1-MSW002-0-041216* 0.227 40 1,70 30 NR NR C056457 OL2-F01-iP W 004-0-041215 ND 30 ND 30 6.80 30 C0056457 Rep DL2-F01-i PW004-0-041215* ND 30 ND 30 6.80 30 C056458 DOU-F01-MSW03--041216 ND 30 10.4 30 NR NR CQ056458 Rep DOU-F01-MSW03-0-41216* ND 30 10.8 30 NR NR 00056459 DL3-F01-IPW004-0-041130 ND 30 ND 30 7.92 30 C0056459 Rep DL3-F01-IPW4--041130* ND 30 ND 30 7.52 30 C0056460 DOU-F01-MSW001-0-041216 7.08 30 15,4 30 C0056460 Rep DOU-F01-MSW001-0-041216* 7.12 30 14.2 30 NR NR NR NR C056461 DOU-F01-MSW004-0-041216 4.72 40 21.7 30 C0056461 Rep DOU-F01-MSW004-0-041216* 4.68 40 22.5 30 NR NR NR NR C056462 DOU-F01-MSW005-0-041216 0,364 30 1.34 30 C0056462 Rep DOU-F01-MSW005-0-041216* 0.36 30 1.41 30 NR NR NR NR C0056463 DOU-F01-MS F005-0-041218 1.41 30 8.40 30 C0056463 Rep DOU-F01-MSFQ05-0-041216* 1.43 30 8.44 30 NR NR NR NR C056464 DL3-FO1-1PW0O2--41130 ND 30 ND 30 12.0 50 C0056464 Rep DL3-F01-1PW002-0-041130* ND 30 ND 30 11.5 50 C0056465 DL3-F01-IPW003-0-041130 ND 30 ND 30 5.44 40 C0056465 Rep DL3-F01-IPW0Q3-0-41130* ND 30 ND 30 5,28 40 C0056466 DOU-P01-MSF003-0-041216 1.72 30 6.56 30 NR NR C056466 Rep DOU-F01-MSF003-0-041216* 1.73 30 6.68 30 NR NR C0056467 DBC-F01-MSF005* 1-041216 0.220 30 1.79 30 NR NR 00056467 Rep DSC-F01-MSF005-1-041216* ND 30 1,76 30 NR NR C056468 CGQ56468 Rep DBC-FG1-MSF005-0-41216 DBC-F01-MSF005-0-041216* ND ND 30 1.68 30 30 1.59 30 NR NR NR NR C056469 DOU-F01-MSF02-0-041216 0.888 30 4.56 30 NR NR C0056489 Rep DOU-F01-MSF002-0-041216* 0.872 30 4.56 30 NR NR C0056470 DL3-F1-IPW001-0-041130 ND 30 0.246 30 9.04 30 C0056470 Rep DL3-F1-IPWG1-0-041130* ND 30 0.219 30 8.80 30 'Laboratory Duplicate ND = Not detected at or above Limit o f Quantitation which is 0.2 ng/g (wet weight}. NR = Not reported due to quality control result failures. Exvgen Research Page 28 of 132 Interim Report #10-Analysxs of Fish and Clam Samples Exygen Study No.: P000I13I Table I. Summary of PFBS, PFHS and PFOS in Fish Samples (continued) Exygen ID C056471 CG56471 Rep C056472 C0056472 Rep C0056473 CQQ56473 Rep C056474 C0056474 Rep C0056475 C056475 Rep C0056476 C0056476 Rep C0056477 C0056477 Rep C056478 C0056478 Rep C0056479 C056479 Rep Client Sample ID C4 Sulfonate PFBS Paffiuorobutanesutfonate Anaiyte Assessed Found Accuracy {ng/g) wet wt. (+/- %) C6 Sulfonate PFHS PerRuorohexanesutfonate Anaiyte Assessed Found Accuracy (ng/g) wet wt. (+/- %) C8 Sulfonate PFOS PerESuorooctanesuffonat# Analyte Assessed Found Accuracy (ng/g) wet wt. f+F %> DOU-FO1-MSF002-1 -041216 DOU-F01-MSF002-1-041216* DOU-F01-MSF004-0-041216 DOU-F01-MSF004-0-041216* DOU-F01-MSF001-0-041216 DOU-F01-MSF001 -0-041216* DBC-F01-MSF03-0-G41216 DBC-F1-MSF003-0-041216* DBC-F01-MSF04-0-041216 DBC-F01-MSF04-0-41216* DMC-F01-MSF01-0-041216 DMC-F01-MS F001-0-041216* DL1-F01- IPW005-0-041216 DL1-F01-IPW005-0-041216* DL2-F01-IPW003-0-041201 DL2-F01-I PW003-0-041201* DL3-F01-1PWGG5-0-41201 DL3-F1-I PW005-0-041201 * 0.840 0.860 0.792 0.816 2.18 2,28 ND ND ND ND ND ND 0.253 0.249 ND ND ND ND 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 4.68 4.72 2.35 2.22 10.9 10.7 2.09 2.09 3.87 3.77 0.420 0.456 0.644 0.644 ND ND ND ND 30 30 40 40 30 30 30 30 30 30 30 30 30 30 30 30 30 30 NR NR NR NR NR NR NR NR NR NR NR NR 68.8 67,6 7.48 7,44 4.88 4.72 NR NR NR NR NR NR NR NR NR NR NR NR 30 30 40 40 30 30 ' Laboratory Duplicate ND = Not detected at or above Limit of Quantitation which is 0.2 ng/g (wet weight). NR = Not reported due to quality control result failures. Exygen Research Page 29 of 132 Interim Report #10-Analysis of Fish and Clam Samples Exygen Study No.: P001131 Table II. Summary of FFOS in Re-extracted Fish Samples Exygen ID C48151 C0048152 C48153 C048154 C0048155 C0048158 C048160 C0049450 C0049451 C049452 C0049453 C0049454 C0049455 C0049458 C49458 C0049459 00049460 C0049461 C049462 C0049463 C0049465 C0049466 C049467 C0049469 C0049471 C0049472 C lient S am ple ID D S C -F 1-IP W Q 01-0-041021 D BC-F01-IPW 002-0-041021 D B C -F 0 1-IP W 0 0 3 -0 - 4 1021 DO U-F01-PF001-0-041021 D O U - F 0 1-IP F 0 0 2 -0 -0 4 1 021 D O U -F Q 1 -IP F Q 05-0- 4 1021 D O U -F 0 1-SP W 0 0 2 -0-041 021 DBC-F01-M SW 001 -0-041104 DBC-F01-M SW 002-0-041104 D B C -F01-M S W 003-0-041104 D 8C -F01-M SW 004-0-041104 D BC-F01-M SW 005-0-041104 D B C -F 0 1-MSFQQ1-0 -0 4 1 1 0 4 DBC-F01-M SF002-0-041104 DBC-F 1-IPF005-0-041104 DBC-F01-I PW 004-0-041104 D B C - F 0 1-I P W 0 0 5 -0 -0 4 1 1 0 4 DO U-F 1-IPW 003-0-041104 D U -F 01 -I P W 0 0 4 -0 -0 4 1 1 0 4 DOU-F01-IPW 005-0-041104 DL1-F01-M SW 001 -0-041104 D L 1-F 0 1-M S W 0 2 -0 -0 4 1 104 D L 1- F 0 1-M S W 0 0 3 -0 -0 4 1104 D L1-F01-MSW5--041104 D L 1 -F 01 -M S F 0 2-0 -04 1104 D L1-F01-M SF003-0- 41104 NR = Not reported due to quality controf result failures. Exygen Research C8 Suffonate PFOS Perffuorooctanestilfonate Analyte Found Assessed (ng/g) Wet Weight A c c u ra c y {+/- %} 2080 30 3330 30 1660 50 596 30 636 30 544 30 1210 9480 40 40 7520 30 12600 30 3440 30 5240 30 5240 30 4760 608 1230 30 30 40 2620 30 NR NR 1740 30 2130 30 968 30 708 30 698 30 604 30 568 30 432 30 Page 30 of 132 Interim Report #10-Analysi$ of Fish and Clam Samples Exygen Study No.: P01131 Table II. Summary of PFOS in Re-extracted Fish Samples (continued) Exygen ID C049474 C0049498 C0049499 C056456 C0056458 C056460 C0056461 C0056462 C0056463 C0056466 C0056467 C0Q56468 C0056469 C0Q56471 00056472 C0056473 C56474 C0G56475 C0056476 C0048148 C0048150 CQ48156 C0048157 C0048159 C0049473 Client Sam ple ID DL1-F01-MSFQQ5-Q-041104 DMC-F01-M SW 005-0-041105 DMC- F1 -MS F002--041105 DO U -F01-MSW 002-0-041216 DOU-F1-M S W 003-0-041216 DOU-FQ1-MSW 0 0 1-0-041216 D O U -F01-M S W 004-0-041216 DOU-F1-MSW05-0-041216 D O U -F01-M SF005-0-041216 DOU-F01-MSF003-0-41216 DBC-F01-MSF005-1 -041216 D B C -F 01-M S F 00 5 -0 -0 4 1 2 1 6 DU-F01-MSF02-0-041216 DOU-F1-M S F002-1-041216 DOU-F01-MSF004-0-041216 DOU-F01-MSFQ01-0-041216 DBC- F01-MSF003-0-41216 D B C -F 1-M S F 0 4 -0 -0 4 1216 D M C -F 01 -M S F 0 0 1-0 -04 1 216 DBC-F01-IFFQ01-0-04121 DBC-F01-IPF003-0-041021 DOU-F01 -I PF003-0-041021 DQU-F01-1PF004-Q-04121 DOU-F01 -IPW1--41021 DL1-F01-M SF004-0-041104 NR = Not reported due to quality control result failures. C8 Sulfonate PFOS Pcrfluorooctanesulfo nate Analyte Found Assessed (ng/g) Wet Weight A ccuracy (+/- %) 740 30 560 30 620 30 28200 60 4040 NR 30 NR 5120 30 496 30 720 30 848 NR 8960 640 692 30 NR 50 30 30 1820 30 2070 30 4640 30 9280 840 50 30 384 30 293 30 359 30 424 30 387 30 348 30 Exygen Research Page 31 of 132 interim Report #10-Analysis of Fish and Clam Samples Exygen Study No.: P0001131 Table III. Summary of PFBS5PFHS and PFOS in Clam Samples Exygen iD Client Sampie iD C4 Sulfonate PFSS Perftuorobutanesulforiate Analyte Assessed Found Accuracy (ng/g) wet wt. .....(+E%)..... C6 Sulfonate PFHS Perfluorohexanesulfonate Analyte Assessed Found Accuracy (ng/g) wet wt. M - %) C8 Sulfonate PFOS Perfiuorooctanesulfonate Analyte Assessed Found Accuracy (ng/g) wet wt. (+/- %} C0056480 DL3-I01-CFW001-0-041216 ND 40 ND 40 1.82 50 C0056480 Rep DL3-01-CFWQ01 -0-041216* ND 40 ND 40 1.82 50 CQ056481 DMC-i01-CFW001 -0-041216 ND 30 ND 30 6.16 50 C0056481 Rep DMC-101-CFW001 -0-041216* ND 30 ND 30 6.12 50 C0056482 DL2-101-CFW001-0-041216 ND 30 ND 30 1.81 50 C0Q56482 Rep DL2-I01-CFWQ1-0-G41216* ND 30 ND 30 1.77 50 C0056483 DBC-i01-C FW 001-0-041215 NO 30 ND 30 4.48 30 C0Q56483 Rep D8C-IQ1-CFW01 -0-041215* ND 30 ND 30 4,44 30 C0056484 DL1-I01-CF W 001-0-041215 ND 30 ND 30 8.80 40 00056484 Rep DL1-I01-CFW001-0-041215* ND 30 ND 30 8,84 40 C0056485 DOU-I01-CFW001-0-041216 C0056485 Rep DOU-I01-CFW001-0-041216* ND ND 30 0.202 30 30 0.205 30 11.4 10.9 50 50 Laboratory Duplicate NO = Not detected at or above Limit of Quantitation which is 0.2 ng/g (wet weight). Exygen Research Page 32 of 132 Interim Report # 10-Analysis of Fish and Clam Samples Exygen Study No.: P0001131 Table IV. Matrix Spike Recovery PFBS, PFHS and PFOS m Fish Samples Sample Description C4 Sulfonate P F B S _______ ._______C6 Sulfonate PFHS________ C8 Sulfonate PFOS Amount Amt Found Amount Amt Found Amount Amt Found Amount Spiked In Sample Recovered Recovery in Sample Recovered Recovery In Sample Recovered Recovery iogfg) (ng/g)wetwt. ingig)wetwt. ...m ... (ng/g)wetwt, (ng(g)wetwt. ....(%)... (rvgyg)wetwt. (ng/g)wetwt, {%) DL3-F01"IPFQ01--G41Q21 {C004S1265pkC>10ppt>) DL3-FGMPF0Q1-0-041021 (COW0126SpkF, 100ppb) 10 0,309 100 0.309 6,56 62,4 63 62 ND 8.28 83 5.16 ND 73.2 73 5.16 12,6 74 78.4 73 DL3-F01-PF002-0-041021 (C004B127SpkDr10ppb) DL3-F1-IPF002-0-041021 (CCKW8127SpkG, 100ppb) 10 0,265 6,52 63 100 0.265 69.6 89 ND 7,48 75 4.16 ND 79.6 80 4.16 10,3 61 80.8 77 DMC-FQ1-IPF001-0-041021 (00048120Spk, 10ppb) 10 0.330 4.48 42 DMC-FQt-IPFG1-0-041021 (C0048128SpkH, 100ppb) 100 0.330 66.4 66 ND 5.24 52 7.40 ND 74.8 75 7.40 9.96 26 81.6 74 DMC-F01"1PFGG2-0-041G21 (C4B12SSpkC, 1Qppb) 10 0.712 7.68 70 DMC-F01-IFFQQ2-0-41G21 {C0O4l2aSpkH(1ppb) 100 0,712 78.0 77 ND 9.60 96 14.1 ND 96.0 96 14.1 29.6 155 104 90 DMC-F1-PW01-0-041021 (C0G4&13OSpkD, 10ppb) 10 1.08 7.60 65 ND 7,84 78 70.0 71.6 * DMC-FHPW01*0^041021 fC004ef30SpkL 10Gppb) 100 1.08 64.8 64 ND 80.0 80 70.0 131 61 DMC-FO-PW0O2-O-O41O21 (C004St31 SpkE. 10ppb) 10 0.968 6.76 58 DMC-F01-IPW002-0-041021 (C0048131SpkJ, 100ppb) too 0.968 60,8 60 ND 7.96 80 34,4 ND 74.8 75 34.4 44.8 . 101 67 DMC-F01-MSW01-0-041021 (C0048132Spk, 10ppbSpike) 10 0,218 9.76 95 0.524 9.44 89 DMC-F-MSW01-0-041021 (C00481MSpkF,100ppbSpike) 100 0,218 81,2 81 0.524 93,2 93 270 270 270 * 375 105 DMC*F01-M$W002-0"041021A (0)040133SpkG. 50ppbSpike) 50 NDA 45.2 90 DMC-FO1-MSWGO2-O-041O2tA (C0048133SpkH,500ppbSpike) 500 NDA 432 86 NDA 52,0 104 NDA 482 96 530 530 470 * 924 79 DL2-F014PF001*0^041021 (C04B134SpkF, 10ppbSpike) DL2-F01-IPF001-0-041021 (COO40134SpkK, 100ppbSpike) Oo 10 0.660 5.80 0.660 46.4 51 46 ND 6.56 66 3.45 ND 58.8 59 3,45 8.36 49 56,4 53 DL2-F01-IPF02-0-41021 (0046135SpkG, 10ppbSpite) 10 0.816 5.64 48 DE2-FG14PFQ02-G-G41021 (00048135SpkLh100ppbSp&e) 100 0.816 64.8 64 ND 6.40 64 4.72 ND 78.8 79 4.72 8.56 38 72.4 68 '`Sample residue exceeds the spiking level significantly; therefore, an accurate recovery value cannot be calculated A 1,0 g of sample used for extraction instead of 5,0 g, ND = Not detected at or above Limit of Quantitation which is 0,2 ng/g fwei weight), NDA = Not detected at or above Limit of Quantitation which is t .0 ng/g (wet weight), HR = Not reported due to quality control result failures. Re-analysts results are shown in Table tl. Note: Since this summary table shows rounded results, recovery values may vary slfghiiy from the values In the raw data. Exygen Research Page 33 of 132 Interim Report #IO-Analysis of Fish and Clam Samples Exygen Study No.: P0001131 Table IV. Matrix Spike Recovery PFBS, PFHS and PFOS in Fish Samples (continued) Sample Description DL2-FC14PF003~G~041021 (CG048?36 Spk C, 10 ppb Sp&e) DL2-FG1-IPF003-0-041021 (000481C6 Spk H, 100 ppb Spike} DL2-F01-SPFG04-0-041021 (00048137 SpkO,lOppb Spike) DL2-FG1-IPF004-G-041021 (C0048137 Spk t, 100 ppb Space) DL2-FG1-1PFOO5-G-041021 (CG04813& Spk E, 10 ppb Spike) DL2-F01 -IPF0O5~Q~G41021 (C0G48136 Spk J, 100 ppb Spike) DL2-F01-SPWOQ1-0-041021 (C0G48139SpkF, 10 ppb Spite) DL2-FG1-SPW001 -0-041021 (CQ048139 Spk K. 10Qppb Spike) DL2-F01-MSF001 -04341021 (00048140 Spk G. 10 ppb Spike) D L 2 -F 0 1 - M S F 0 0 1 - 0 4 K 1 021 (C004614G Spk l r 100 ppb Spike) DL2-F01-MSF002-0-041021 (0048141 Spk C, 10 ppb Spike) DL2-F01-MSF002-0-041021 (C0043U1 Spk H, 100 ppb Spike) DL2-F01-MSFOO3-0-O41021 (C0IM8U2 spkD, 10 ppbSpike) DL2-FQ1-MSF003-G-041021 00048142 Spk I. 100 ppb Spite) DL.2-F01 -MSF004-0-G41021 (C0048143 Spk E. 10 ppb Spike) DL2-F01 "M$FQG44>041021 (000431-43 Spk J, 100 ppb Spike) D L 1 -F 0 1 - IP F 0 0 1 - 0 - 0 4 1021 (CO04S144 Spk F, 10 ppb Spike) DL1-F01-IPF001-0-041021 (C0048144 Spk K. 100 ppb Spike) DL3-F01-SPF003-0-041021 (C0048145 Spk G, 10 ppb Spike) DL3-F01-IPF003-0-041021 (C0048145 Spk L, 100 ppb Spike) Amount Spiked {ng/g} C4 Sulfonate PFBS_______________ C6 Sulfonate PFHS________________ C8 Sulfonate PFOS Am t Found in Sample (ng/gSwetwt. Amount Recovered Recovery (nip)wetwt. ..... m .... Am i Found in Sample (ng/g) wet vrt. Amount Recovered (ng/g) wetwt. Recovery <%} Amt Found in Sample (ng/g; wet wt. Amount Recovered Recovery (ng/g) wet wt. ...... w ...... 10 0.247 100 0.247 7.00 62.8 68 63 NO 7.96 80 2.01 ND 73.2 73 2.01 9.20 74.0 72 72 10 0.346 100 0.346 6.84 67.6 65 67 ND 7,60 76 ND 76.8 77 10 0.338 6,88 65 ND 7.68 77 100 0.338 68.4 68 ND 80.8 81 10 0,326 100 0.326 7.04 66.0 67 66 ND 7.60 76 ND 77.2 77 10 0,333 100 0.333 10 0.408 100 0.408 10 0,418 100 0.416 7.36 64.8 6.88 61.6 6.52 64.8 70 64 65 61 61 64 NO 8.20 82 ND 77.6 78 ND 7.48 75 ND 68.0 68 ND 6.76 68 ND 70.8 71 10 0.345 100 0.345 7,08 71,6 67 71 ND 7.56 76 ND 81.2 81 10 0.432 7.44 70 0,261 8,16 79 100 0.432 67.2 67 0.261 76.4 76 10 0.448 7.20 68 ND 8.08 81 100 0,440 63.2 69 ND 81.6 82 4,52 4.52 2.46 2.46 5.88 5.88 77.2 77,2 94.8 94.8 41.6 41,6 83.6 83.6 33.4 33,4 3.55 3.55 11.3 80.4 9.68 82.0 13.9 85.2 76.0 158 101 156 41,2 104 91.2 167 40.0 106.0 10.8 80.0 68 76 72 SO 60 79 * 81 * 61 . 62 * 83 * 73 73 76 ` Sample residue exceeds the spiking level significantly; therefore, an accurate recovery value cannot be calculated ND = Not detected as or above Limit of Quantitation which is 0.2 ng/g {wes weight). NR * Not repotted due to duality control result failures. Re-analysis results are shown in Table II. Note; Since ibis summary table shows rounded results, recovery values may vary slightly from the values in the raw data. Exygen Research Page 34 of 132 Interim Report # 10-Analysis of Fish and Clam Samples Exygen Study No.: P0001131 Table IV. Matrix Spike Recovery PFBS, PFHS and PFOS in Fish Samples (continued) Sample Description L3-F1-IPF004-0-041021 {CG4S14&Spk C< 10 ppb Spike} DL3-F1-IPFG04-0-041021 (C0048146 Spk H, 100 ppb Spike) DL3-F1-1FF005-0-041021 (C004SU7 Spk Dl10 ppb Spike} DL3-F01- IPF005-0-41021 {0004614? Spk l, 100 ppb Spike) DBC-F01-1PF001-0-041021 (CC48148 Spk , 10 ppb Spike) D8C-F1-1PF01 -0*041021 fC04146 Snk J, 100 ppb Spike) DBC-F01-IP F002-0-041021 {C048149 Sf* f , 10 ppb Spike) DBC-F01-IPF002-0-041021 (CGQ48149 Spk K, 100 ppb Spike) DBC-F01-IPF003-0-041021 (C0G48150 spk G, 10 ppb Spike) DBC-F01-1PF03--041021 (C004150 spk L 100 ppb Spike) DBC-F01- 1FW01-0-041021 (C0048151 Spk C. 10 ppb Spike) DBC-F01-1PW001-0-041021 (C0048151 Spk h, 100 ppb Spike) D B C -F 0 1 4 P W 0 0 2 -0 -Q 4 1021 (C048152 Spk 0 , 10 ppb Spike) DBC-F01-1PW002-0-041021 000*8152 Spk 1,100 ppb Spike) D B C -F 0 1 -IP W 0 0 3 -0 -0 4 1 021 {000*8153 Spk E, 10 ppb Spike) DBC-F01-l PW003-0-041021 (C0048153 Spk j, 100 ppb Spike) OOU-F01-1PF001 -0-041021 (C0048154 Spk F, 10 ppb Spike) DOU-F01-1PF001-0-041021 00048184 Spk K. 100 ppb Spike) D O U -F 0 1-1 PF0 02-0-0 41021 CO4S155 Spk G. 10 ppb Spate) D O U -F 0 1-1PF002-0-0 41021 (G0048155 Spk L, 100 ppb Spike) DOU -F01- iPF03-0-041021 00048156 Spk C, 10 ppb Spike) DOU-F01-1PFGQ3-G-G41021 (C415Spk H, 10ppb SpikeI Amount Spiked (ng/q) C4 Sulfonate PFBS_______________ CS Sulfonate PFHS________________ C8 Sulfonate PFOS Amt Found Amount Amt Found Amount Amt Found in Sample Recovered Recovery in Sample Recovered Recovery in Sample fng/g) wet wi. (ng.'g)wet wi. (%) (rtg/g) wet wt. (ng/g) wet wt. (%) fnp/g) wet wt, Amount Recovered {pg/g)wetwt. Recovery {%} 10 0.784 6,84 61 ND 10.6 106 0.382 8.96 86 100 0.784 69.6 69 ND 88.4 88 0.382 81.2 81 10 1,53 5,44 39 ND 9,16 92 5.48 11.7 62 too 1,53 60.4 59 NO 93,2 93 5,48 86.8 81 10 1.34 6,24 49 0,692 10.8 101 100 1.34 56,4 55 0,692 86,8 86 10 1,43 5,84 44 ND 9,36 94 100 1.43 59.2 5B ND 88.0 88 NR NR 110 110 NR NR NR NR 113 . 189 79 10 0.996 6.72 57 0,644 10,4 38 100 0.996 60.8 60 0,644 88.8 88 NR NR NR NR NR NR 10 0,864 7.08 62 2.21 9.68 75 100 0.864 70.0 69 2.21 88.0 86 NR NR NR NR NR NR 10 0.700 100 0.700 7,56 69.6 69 69 9,12 9,12 18.2 96,0 91 87 NR NR NR NR NR NR 10 0.736 100 0.736 8.32 70.8 76 70 6,76 6.76 16.5 90.8 97 84 NR NR NR NR NR NR 10 4,36 4.36 10.9 76.8 65 72 1.02 9.48 85 1.02 86.0 85 NR NR NR NR NR NR oo 10 6,76 100 6,76 13.1 76.0 63 0.792 9.72 69 0.792 60.0 89 79 NR NR NR NR NR NR 10 5.88 100 5.88 12,8 81.2 69 75 1.02 10.6 96 1.02 88.4 87 NR NR NR NR NR NR ` Sample residue exceeds the spiking ievei significantly: therefore, an accurate recovery value cannot be calculated ND = Mot detected at or above Limit of Quantitation which is 0.2 ng/g (wet weight), NR = Not reported due to duality control result failures. Re-analysis resuite am shown in Table n. Note: Since this summary table shows rounded results, recovery values may vary slightly from the values in the raw data. Exygen Research Page 35 of 132 Interim Report # 10-Analysis of Fish and Clam Samples Exygen Study No.: P0001131 Table IV. Matrix Spike Recovery PFBS, PFHS and PFOS in Fish Samples (continued) Sample Description DOU-FQ14PF0G4-G-G41021 jC004S157 Spk D. 10 ppb Spike) DOU-F014PF004-0-041021 (C<xwai57 S pk i, 10 0 p p b S p ike ) DU-F01dPFQG5*CK)4'i21 (C0G4158 S pk E> 10 ppt> DOU-F01-IPF005-0-041021 (COfWBfSe Spk J . 100 ppb s p ik e ) DU-F01-IPW01 -0-041021 (CQ481S9 Spk F, 10 ppb S pike) DOU-F014PW001 -0-041021 (C D04S159 S pk K, 100 p p b S p ike ) DOU -FQ1--IPW G02-0-41021 (CGG48160 S p k G , 10 p p b S p tkb ) D Q U - F 1 -IPW002-Q-041021 (C004S160 Spk L 100 ppb S pike} DL2-F01- M S W 1-0-041021 (C04B161 S pk l, 10 p p b S pike) D L 2 -F 0 1 -M S W 0 0 1-0*041021 1C04161 S pk J f 100 p p b S p ik e ) DL2-F1- M S W 0 0 2 -0 -0 4 1021 (00048162 Spk K f 10 ppb Spike) DL2 -F01 -MSW 002-0-041021 (C0O4S162 S p k L, 100 p p b s p ik e } OL2-F01 -MSW003-0-041021 (CQ4S163 Spk C, 10 ppb S pike) D L 2 -F 0 1 -M S W 0 0 3 -0 -0 4 1021 00048163 Spk 0 , to o ppb Spike) DL2 -F1-MSW 004-0-041021 {C 0046164 Spk E, 10 ppb S pike) DL2-F 01-M S W 0 04-0- 41021 (C 0048164 Spk F . 100 ppb Spike) D L 2 -F 0 1 -M S W 0 0 5 -0 -0 4 1021 (C004818S S pk G , 10 ppb S pike) D L 2 -F 0 1 -M S W 0 0 5 -0 -0 4 1021 (C004165 Spk H f 100 ppb Spike) DBC-F01-MSW0G1 -0-041104 (C004945Q Spk C, 10 ppb Spike) DSC-F144SWG01 *0*041104 (C0G46450 Spk 0, 100 ppb Sptfce) Amount Spiked (9/9) C4 Sulfonate PFBS_______________ CS Sulfonate PFHS Amt Found in Sample Irtp/g) w et wt. Amount Recovered fng/gj w et wt. Recovery {% ) Amt Found in Sample fn g /jj} w et wt. Amount Recovered fn g /g jw e t wt. Recovery (% ) CS Sulfonate PFOS Am t Found in Sample fng/g) w et wt. Amount Recovered jnp/gS w e tw t. Recovery 10 8,12 100 6,12 13,2 76.4 71 70 2.30 2.30 12.0 86.0 97 84 NR NR NR NR NR NR 10 4.56 100 4.56 12.2 76 0.956 11.2 102 76.0 71 0,956 85,6 85 NR NR NR NR NR NR 10 2.89 to o 2.89 10.2 72.4 73 0.948 10.7 70 0.948 83.6 98 83 NR NR NR NR NR NR t o 18.4 34.4 160 3.69 13.2 95 O O 18.4 94.8 76 3.69 89.2 86 NR NR NR NR NR NR 10 0.203 9,32 91 ND 8.68 87 120 100 0.203 76,8 77 ND 85.2 85 120 126 177 57 10 ND 100 ND 10 ND 100 ND 10 ND 100 ND 9,68 86.4 9.24 81.2 7.56 82.0 97 86 92 81 76 82 ND 9.12 91 ND 93.6 94 102 102 ND 9.08 91 92.4 ND 95.2 95 92.4 ND 7.28 73 88.0 ND 93,6 94 88.0 110 185 103 182 75.6 166 * 83 * 90 . 78 10 N D 7.96 80 ND 8.08 81 168 too ND 90.8 91 ND 103 103 168 135 . 234 66 10 0,888 100 0,888 10.3 90.4 94 90 8.32 8.32 17.6 93 113 105 NR NR NR NR NR NR *Samp!e residue exceeds the spiking level significantly; therefore, an accurate recovery value cannot be calculated NO = Not detected s i or above lim it of Quantitation which is 0.2 ng/g (wet weight). MR * Mot reported due to quality control result failures. Re-analysts results are shewn in Table If, Note; Since this summary table shows rounded results, recovery values may vary slightly from the values In the raw data. Exygen Research Page 36 of 132 Interim Report # 10-Analysis of Fish and Clam Samples Exygen Study No.: P0001131 Table IV. Matrix Spike Recovery PFBS, PFHS and PFOS in Fish Samples (continued) Sample Description D B O F 1 -M S W C 2 -0 -0 4 1 104 {C0043451 Spk e, 10 ppb Spike) DBC-FO1-MSW 002-0-041104 (00043451 Spk F, 100 ppb Spike) DE3OF01-MSW3-0-041104 {G4&452 Spk Gf fO ppb Spike) DBC-F1-MSW003--041104 (00049452 Spk H, 100 ppb Spike) O B C -F 0 1 -M S W 0 0 4 -0 -0 4 1 104 {CO0494S3 Spk 1.10 ppb Spike) DBC-F01-MSWO4--O41104 (C0049453 Spk J, 100 ppb Spike) DBC-F01-MSWG05-0-041104 (CQ049454 Spk K, 10 ppb Spike) DBC-F01- NSSWQO5-0-041104 (CG049454 Spk L, 100 ppb Spike) DBC-F01-MSF001 -0-041104 (C0Q49455 Spk M, 10 ppb Spike) DBOF1 4SF1 *0*041104 C0049455 Spk N, 100 ppb Spike) DBC-F1-MSF002-0-041104 (C00494S6 Spk 0, 10 ppb Spike) OBC-F01-MSF02-0-41104 (CD04945Spk Ph100 ppb Spite) DBC-F01 -IPF004-G*041104 (C04457 Spk: C, 1ppb Spinel DBC-F1-IPF04-0-G41104 (CCM)49457 Spk 0,100 ppb Spike) D 8 C -F 0 1 4 P F 0 0 5 - 0 - 0 4 1 104 (C4945BSpk E, 10 ppb Spike) DBC-F01-IPF006-0-041104 (C0D4945S Spk Fr 100 ppb Spite) D B C *F 0 1 4 P W 0 0 4 -0 *0 4 1 104 C0049459 Spk G, 10 ppb Spfee) DBC-F01-t PW 004-0-041104 (C0049459 Spk H. 100 ppb Spite) D B C -F 0 1 -IP W 0 0 S -0 -0 4 1104 (C0Q4946G Spk 1,10 ppb Spite} D B C -F 0 1- IP W 0 0 5 -0 -0 4 1 104 {C004946 Spk J, 100 ppb Spite) Amount Spiked io g /g ) C4 Suifonate PF8S_______________ C6 Suifonate PFHS________________ C8 Sulfonate PFOS Am t Found Amount Amt Found Amount Amt Found n Sampie Recovered Recovery in Sample Recovered Recovery in Sampfe fog.'j) wetwt. (ng/g) wet wt. <%) (ng/g) wet wt. (ngig)wetvrt. (%> (neig) wet wt. Amount Recovered fng/g) wet Recovery <%) to 0.868 100 0.868 9.96 94,8 91 94 3,43 3,43 12.8 94 113 110 NR NR NR NR NR NR 10 0.860 100 0.860 9.92 88.0 91 85 3.62 3.62 13.6 100 104 100 NR NR NR NR NR NR 10 0.884 10D 0.884 9.96 97.2 91 96 9.72 9.72 19.5 98 124 114 NR NR NR NR NR NR 10 0.832 100 0,832 8.20 91.6 74 91 2.77 2 .7 7 10.2 74 108 105 NR NR NR NR NR NR 10 0.820 100 0.820 9.60 88,4 88 88 3.58 3.58 13.7 101 106 102 NR NR NR NR NR NR 10 0,768 100 0,768 9,92 90.8 92 90 1.94 1.94 10 0.504 100 0,504 10.5 94.8 100 0.584 94 0.584 12.5 106 102 100 11.6 110 106 105 NR NR 142 142 NR NR NR NR 152 * 254 112 10 1.60 100 1.60 10.4 90.8 88 89 2.15 2.15 12.2 101 105 103 NR NR NR NR NR NR 10 0.484 100 0.484 9.72 87,8 92 87 3.28 3.28 13.0 97 104 101 NR NR NR NR NR NR 10 0,572 100 0.572 10.4 92.8 98 92 2.10 2.10 13.2 111 102 100 NR NR NR NR NR NR ` Sample residue exceeds die spiking level significantly; therefore, an accurate recovery value cannot be calculated ND = Not detected at or above Limit of Quantitation which is 0,2 ng/g (wet weight). NR = Not reported due to qualify control result faifures. Re-analysis results are shown in Table It. Note: Since this summary table shows rounded results, recovery values may vary slightly from the values in the raw data. Exygen Research Page 37 of 132 Interim Report # 10-Analysis of Fish and Clam Samples Exygen Study No.: F0001131 Table IV. Matrix Spike Recovery PFBS, PFHS and PFOS in Fish Samples (continued) Sample Description DOU-FG1-SPWG03-0-G41104 (CQ04S461 Spk It, 10 ppb Spike; DOU-FG1-1PW0O3-O-O411Q4 {C0049461 Spk L, 100 ppb Spike) D O U -F 0 1 -IP W 0 0 4 -0 -0 4 1 104 (C0049462 Spk C. 10 ppb Spike) DO U-F01-1PW0G4-0-041104 (C004S462 Spk D, too ppb Spike) DOU-F01-IPW0G5-G-041104 (C0049463 Spk E, 10 ppb Spike) DOU-F01 -IPW0Q5-G-O41104 00049463 Spk F, 100 ppb Spike) D I2 -F 0 1 -1 P W 0 0 2 -0 -0 4 1104 (C0049464 Spk G, 10 ppb Spike) D L 2 -F 0 1 4 P W 0 0 2 -0 -0 4 1 104 (C0049464 Spk H, 100 ppb Spike) D L1-FG1 'MSW001-0-041104 (C0049465 Spk k 10 ppb Spike) DL1-F01-MSW001 -04341104 {030494&5 Spk Jr 100 ppb Spike) DL 1-F01-MSW002-0-041104 {C0048466 Spk K, 10 ppb Spike) DL1-F01-MSWOO2-0-O41104 (C004S466 Spk L, 100 ppb Spike) DL 1-FG1-MSW003-0-041104 (C004&467 Spk C, 10 ppb Spike) DL1-F01-MSW0G3-0-041104 (C0049467 Spk D. 100 ppb Spike) DL 1' F01-MSWC04-0-041104 {C004&468 Spk E, 10 ppb Spike) O L1-F0 1 -M S W 0 0 4 -0 -C 4 1 104 (C o o t9466 Spk F. too ppb Spike) DL 1-F01-MSW005-043411G4 fC0G494e& Spk Gr 10 ppb Spike) D L 1 -F 0 1 -M S W 0 0 5 -0 -0 4 1 104 (C0043469 Spk H. 100 ppb Spike) DL1-F01-MSF001-0-041104 {C0049470 spk 1,10 ppb Spike) DL1-FQ1-MSF001-0-041104 (C0049470 Spk J, 100 ppb Spike} Amount Spiked fhS/Q) 0 4 Sulfonate PFBS Amt Found Amount in Sample Recovered Recovery (ng/gi wet wt. (ng/g) wet wt. .. . (%) C6 Sulfonate PFHS _______ C8 Sulfonate PFOS Amt Found Amount Amt Found in Sample Recovered Recovery in Sample (ng/g) wet wt. (ng/g) wet wt. {%> (ng/g) wet wt. Amount Recovered (ng/g) wet wt. Recovery W ..... 10 3.12 100 3.12 13.1 88,0 100 85 6.00 6.00 16.3 103 107 101 NR NR NR NR NR NR 10 3.49 100 3.49 12.8 88.4 93 85 7,12 7,12 17.4 103 108 101 NR NR NR NR NR NR 10 2.97 100 2.97 10.5 83.6 75 81 5.12 5.12 13.5 84 109 104 NR NR NR NR NR NR 10 1.04 100 1,04 9.48 80.0 84 79 ND 10.4 104 8.20 ND 97.6 98 8,20 14,3 61 124 116 10 1.20 100 1.20 10.6 94 0.896 11.3 104 89.6 8 8 0.896 112 111 NR NR NR NR NR NR 10 1.30 100 1.30 9.76 80.4 85 79 1.72 12.4 107 1.72 100 98 NR NR NR NR NR NR 10 0.560 100 0.560 8,92 85.2 84 85 1,13 1.13 10.2 98.8 91 98 NR NR NR NR NR NR 10 0.908 8.52 76 0.908 9.76 89 100 0,908 84,8 84 0.908 99.6 99 181 181 201 289 108 1 0 1.18 100 1.18 8.80 82.4 76 81 1.18 1.18 10.4 98,8 92 98 NR NR NR NR NR NR 10 ND 100 N D 7.76 78 0,315 9.52 92 76.4 76 0.315 90.8 90 293 293 303 * 394 101 'Sample residue exceeds the spiking level significantly; therefore, an accurate recovery value cannot be calculated NO = Not detected at or above Limit of Quantitation which is 0,2 ng/g {wet weight), NR = Mot reported due to quality control result failures. Re-analysis results are shewn in Table II, Note: Since this summary table shows rounded resuite, recovery values may vary slightly from the values In the raw data. Exygen Research Page 38 of 132 Interim Report # 10-Analysis of Fish and Clam Samples Exygen Study No.: P0001131 Table IV. Matrix Spike Recovery PFBS, PFHS and PFOS in Fish Samples (continued) Sample Description DL 1-FQ1-MS F002-0-041104 (C049471 Spfc K. 10 ppb Spike) D l 1-F01-MSF02-0-G41104 iC004S4?1 Spk t, 100 ppb Spite) DL1-F01-MSF003-0-G41104 (CO049472 Spk C, 10 ppb Spre) D Lt -F01-MSF3-O-041104 (C0048472 Spk D, 100 p i* Spike) DU-F01-MSF004-0-41104 {C004M73 Spk 10 ppb Spike) DL 1-FG1-MSFOG4-0-41104 (0X149473 Spk F. 100 ppb Spike) DL1-F0 MWSF005*0*041104 (C4474 Spk Gr ppb Spike) Ot1 -F01-MSF005-0-041104 {C43474 Spk H, 100 ppb Spike) DL1-F14PW0G1*0*041104 (CO049475 Spk 1,10 ppb Spike) DL1-F0 i-IPW O01-0-041104 (C0Q49475 Spk J, 100 ppb Spike) DL1-FG14FW02--041104 {CO40476 Spli K, 10 ppb Spike) DL1-F01-SPW02-0-041104 (C0049476 Spk L. 100 ppb Spike) DL 1-F01-IPWO03-O-G41104 (C04S477 Spk C, 10 ppb Spike) D t1 -F 01-IPWG03--041104 (C0049477 Spk D, tOOppb Spike) DL1-F01-1PW004-0-041104 (0)049478 Spk E, 10 ppb Spike) DL1-F01 *iPW 004-0-041104 (00043470 Spk F, 100 ppb Spike) DL1-F01-SPF2-0-41104 (C0049479 Spk G. 10 ppb Spike) DL1-F01-IPF002--041104 (C004047 Spk Hr 100 ppb Spike) DL-1-F01-IPF003-0-41104 (C00494S0 Spk f, 10 ppb Spike) DL-1 *F014PF003-0-041104 (CO04948O Spk J, tOOppb Spre) C4 Sulfonate PFBS_______________ C6 Sulfonate PFHS________________ 0 8 Sulfonate PFOS Amount Am t Found Spiked in Sample i...M l . (ng/g) wet wt. Amount Recovered Recovery (ng/g) wet wt. (%) Amt Found in Sample (ng/g) wet wt. Amount Recovered ing(g)wetwt Recovery {%! Am! Found in Sample (ngfg) wet wt. Amount Recovered (rtp/S) wetwt- Recovery (%) 10 ND 100 NO 8.60 86 0.624 10.5 99 84.8 85 0,624 98.4 98 NR NR NR NR NR NR 10 0.203 8.76 86 0.652 10.2 95 100 0.203 84.4 84 0.652 95.2 95 NR NR NR NR NR NR 10 ND 100 ND 8,88 89 0,532 10.4 99 86.8 87 0.532 101 100 NR NR NR NR NR NR 10 ND 100 ND 8.64 86 0.684 10.1 94 82.4 82 0.684 94,0 93 NR NR NR NR NR NR 10 0,772 8,64 79 0,556 9.72 92 100 0,772 84.0 83 0.556 97.6 97 17.4 17.4 26.4 113 90 96 10 0.4S8 8.68 82 0.536 9.96 94 100 0.468 76.4 76 0.536 88.0 87 15,7 15.7 27.4 100 117 84 10 ND 100 ND 10 ND 100 ND 9.08 91 0,209 9.44 92 9.36 90.4 90 0.209 105 105 9.36 8.48 85 0.238 9,80 96 71.6 72 0.238 84.8 85 59.6 59,6 16.7 109 72.0 140 73 100 . 80 10 NO 100 ND 8.00 80 0.222 9.28 91 86.4 86 0.222 98.4 98 11.1 11.1 18.9 78 108 97 10 ND 100 ND 8.80 88 0.265 10.6 103 20.9 78.8 79 0.265 90,4 90 20,9 30.5 109 96 88 ` Sample residue exceeds the spiking level significantly; therefore, an accurate recovery value cannot be calculated NO = Not detected at or above lim it of Quantitation which is 0,2 ng/g (wet weight), NR = Not reported due to quality control result failures. Re-analysis results are shown in Table II. Note: Since this summary table shows rounded results, recovery values may vary slightly from the values In the raw data. Exygen Research Page 39 of 132 Interim Report # 0-Analysis of Fish and Clam Samples Exygen Study No. : POOO1131 Table IV. Matrix Spike Recovery PFBS, PFHS and PFOS in Fish Samples (continued) Sample Description DL-1-FQ1-IPF004-0-041104 (01049481 Spk K, 10 ppb Spike) DL-1 -F01-SPFOO4-O-041104 (C004B481 Spk L. 100 ppb Spike) DL-1 -F01-1PF0G5-G-041104 {C004S482 Spk C. 10 ppb Spike) DL-1 -F014PFO05-0-O41104 (C0049482 Spk D, 100 ppb Spike) DL3-F01-MSW001-0-041105 (00049483 Spk E, 10 ppb Spike) DL3-F01-MSW001-0-041105 {C0049483 Spk F, 100 ppb Spike) DL3-F01-MSWCC2-Q-041105 (CO04S484 Spk G. 10 ppb Spike) D L 3 -F 0 1 -M S W 0 0 2 -0 -0 4 1105 (C0049484 Spk H, 100 ppb Spike) D L 3 -F 0 1 -M S W 0 0 3 -0 -0 4 1 105 (C0049485 Spk 1,10 ppb Spike) DL3-F01-MSW003-G-041105 (C0G49485 Spk J. 100 ppb Spike) DL3-F01-MSW004-G-041105 (CG049466 Spk K, 10 ppb Spike) DL3-FG1-MSW0G4-G-041105 {C0G4S4S6 Spk Lt 100 ppb Spike) D L 3 -F 0 1 -M S W 0 0 5*0*0411 05 (C0049487 Spk C, 10 ppb Spike) O L3-F 0 1 -M S W 0 0 5 -0 -0 4 1105 (00049467 Spk 0.100 ppb Spike) DL3-F01 *MSF001 *0-041105 (C0049488 Spk E, 10 ppb Spike) DL3-F01-MSF001 *0-041105 (C00494S8 Spk F, 100 ppb Spike) DL3-F01-MSF0G2-O-O41105 (C0049489 Spk G. 10 ppb Spike) DL3-FQ1-NSSFG02-0-041105 (C0049489 Si* H, 100 ppb Spike) DL3-FQ1-MSF003-0*041105 (C0049490 Spk 1,10 ppb Spike) DL3-F01-MSF003-0-041105 (C004S490 Spk J, 100 ppb Spike) Amount Spiked <ng/g) C4 Sulfonate PFBS Amt Found in Sample (ng/g) wet wt. Amount Recovered Recovery (ng/g) wet wt. {% ) . C6 Sulfonate PFH$________________ CS Sulfonate PFOS Amt Found Amount Amt Found Amount in SampSe Recovered Recovery in Sample (ng/g) wet wt. n g /g l wet wt. ......( * ) . . fng/g) wet wt. Recovered Recovery (n g /g ) w e t w t. ........ L * 1 ........ 10 ND 100 ND 8.00 84.8 80 85 ND 9,48 95 7.60 ND 98.8 99 7.60 15,8 82 102 94 10 ND 100 ND 7,44 74 0.373 9.00 86 84.8 85 0.373 38.4 98 8.72 8.72 15.9 72 102 93 10 ND 100 ND 8,36 85,6 84 86 ND 9.20 92 139 ND 99,6 100 139 165 * 258 119 10 ND 100 ND 8.44 90.4 84 90 ND 9.92 99 160 ND 106 106 160 150 * 263 103 10 ND 100 ND 10 ND 100 ND 10 ND 100 ND 10 ND 100 NO 10 ND 100 ND 10 ND 100 ND 8.60 88.4 8.48 88,4 9.56 100 9,48 94.8 9.88 89.6 9,32 90.8 86 ND 88 ND 85 0.285 88 0.285 96 0.366 100 0.366 95 ND 95 ND 93 ND 90 NO 93 ND 91 ND 9,56 102 9.68 102 11.1 105 11.4 109 10,4 103 11.0 107 96 102 94 102 107 105 114 109 104 103 110 107 146 146 198 198 173 173 116 116 116 116 75,6 75,6 154 254 225 301 193 246 138 234 96.8 216 92.0 190 * 108 * 103 73 * 118 * 100 . 114 ' Sample residue exceeds the spiking level significantly; therefore, an accurate recovery value cannot be calculated ND = Not detected at or above Limit of Quantitation which is 0.2 ng/g (wet weight). NR = Not reported due to quality control result failures. Re-analysis results are shown in Table il. Note: Since trs summary tabi shows rounded results, recovery values may vary slightly from the values in the raw data. Exygen Research Page 40 of 132 Interim Report #10-Analysis of Fish and Clam Samples Exygen Study No.: P0001131 Table IV. Matrix Spike Recovery PFBS, PFHS and PFOS in Fish Samples (continued) Sampie Description QL3-FQ1-MSF004-0-G41105 (CQ049491 Spk K, 10 ppb Spike) D L 3 -F 0 1 -M S F 0 0 4 -0 -0 4 1105 (C4&491 Spk L 100 ppb Spike) D L 3 -F 0 1 -M $ F 0 0 5 4 > 0 4 1 105 {C0049492 Spk C, 1ppb Spike) D L 3 -F 0 1- M S F 0 0 5 -0 -0 4 1 105 {C049492 Spk 0 , 100 ppb Spike} DMC-F01-1PW0G3-Q-041105 (C00494S3 Spk E, 10 ppb Spike) DMC-F1-IPW 003-0-041105 (C0049493 Spk F. 100 ppb Spike) DMC-FG1-1P W 004 -0-041105 (C049404 Spk G. 10 ppb Spike) DMC-F014PW004-0*041105 (C0049494 Spk H, 100 ppb Spike) D M C -F Q 1 4 F W 0 0 5 -0 -0 4 1105 (C049495 Spk i, 10 ppb Spike} D M C -F 0 1 -IP W 0 0 5 -0 -0 4 1 105 (C0049495 Spk J, 100 ppb Spike) D M C -F 0 1 -M S W 0 0 3 -0 -G 4 1105 (C0049490 Spk K, 10 ppb Spike) DMC-F01 -M$W0G3"(M)41105 (C0049496 Spk L, 100 ppb Spike> DMC-FG t -MSW 004*0-041105 (C0O49497 Spk C, 10 ppb Spike) DMOF01 -M$WGC4-0-G41105 (C0D4949? Spk 0T10ppb Spake) D M C -F 0 1 -M S W 0 0 5 -0 -0 4 1 105 (00049498 Spk E 10 ppb Spike) D M C -F 0 1 -M S W 0 0 5 -0 -0 4 1105 (C0049498 spk F, 100 ppb Spike) DMC- F01- MSF002-0-041105 (C049499 Spk G. 10 ppb Spike) DMC-FQ1-MSF002~G-G41105 (00040499 spk H, 100 ppb Spike) DMC-F01-MSF3-O-G411O5 (C0049500 spk 1,10 ppb Spike) D M C -F 0 1 -M S F 0 0 3 -0 -0 4 1 105 (00049500 Spk J, 100 ppb Spike) Amount Spiked (ng/g) C4 Sulfonate PFBS_______________ CS Sulfonate PFHS________________ C3 Sulfonate PFOS Amt Found in Sample (ng/g) wet wt. Amount Recovered Recovery (ng/g) wet wt. ......i$ y ..... Am i Found in Sample (ng/g) wet wt. Amount Recovered (ng/g) wet wt. Recovery f%> Am t Found in Sample (ng/g) wet wt. Amount Recovered Recovery (rtg/g) wet wt. ......1%)....... 10 ND 9,80 96 ND 11.0 110 130 100 ND 90,4 90 ND 104 104 130 10 ND 8.92 89 ND 10.3 103 81.6 too ND 85,6 86 ND 98.4 38 81.6 10 ND 100 NO 8.28 83 0.386 9.52 91 84.4 84 0,386 99.6 99 61.2 61.2 10 0.300 8,04 77 0.396 9.28 89 68.8 too 0.300 90.4 90 0,396 104 104 68.8 10 NO 100 ND 8.96 90 0.270 10.1 98 64.4 88.4 88 0,270 105 105 64,4 10 ND too ND 9,64 96 0.592 11.0 104 92,4 92 0.592 106 105 354 354 10 0,318 9.52 92 1.90 12,1 102 259 100 0.318 82.4 82 1.90 101 99 259 144 238 97.2 181 78,0 156 107 168 75.6 153 389 464 259 313 108 . 99 . 95 99 * 89 * 110 . 54 10 ND 100 ND 9.16 92 0.980 11.6 106 93.6 94 0.980 118 117 NR NR NR NR NR NR 10 ND 8,36 84 ND 10.4 104 NR 100 ND 97.6 98 ND 104 104 NR 10 ND 8.20 82 ND 10.4 104 56,8 100 ND 87,2 87 ND 110 110 56.8 NR NR NR NR 108 . 194 137 ` Sample residue exceeds the spiking level significantly; therefore, an accurate recovery value cannot be calculated ND = Not detected at or above Limit of Quantitation which is 0.2 ng/g (wet weight). NR s Not reported due to quality control result failures. Re-anaiysis results are shown in Table II. Note; Since this summary table shows rounded results, recovery values may vary slightly from the values in the raw data. Exygen Research Page 41 of 132 Interim Report #10-Analysis of Fish and Clam Samples Exygen Study No.: P0001131 Table IV. Matrix Spike Recovery PFBS, PFHS and PFOS in Fish Samples (continued) Sample Description D M C -F 0 1 -M S F 0 0 4 -0 -0 4 1 105 (03049501 Spit K, 10 ppb Spike) D M C -F 0 1 -M S F 0 0 4 -0 -0 4 1 105 00049501 Spk L. 100 ppb Spite) D M C -F 0 1 -M S F 0 0 5 -0 -0 4 1 105 'CQ049502 Spk C, 10 ppb Spite) DMC-F01-MSFG05-0-041105 (C0049502 Spk D, 100 ppb Spike) DMC-FQ1 -iPF003-0-Q41105 (0049503 Spk E, 10 ppb Spike) DM OF014PF003-0-041105 (C0049503 Spk F. 100 ppb Spite) D M C -F 0 1 -IP F 0 0 4 -0 -0 4 1 105 (CQ049504 Spk Q. 10 ppb Spike) DMC-F01-) PF004-0-041105 {C004950* Spk H, 100 ppb Spite) DMC-F01 -SPFOOS-O-041105 000*9505 Spk 1,10 ppb Spike) DMC-F 014PF005-G-041105 (C004-0505 SpkJ, 100ppb Spite) DL2-FQ1 -MSF005-0-41216 (CGG56354 Spk E, 10ppb Spike) DL2~F014PW005-0-041215 (C0056354 SpkF, 100ppb Spike) D L 2 -F 0 1- IP W 0 0 5 -0 -0 4 1 215 {C0C56455 Spk C, 10ppb Spike) OL2-F01-IPW005-G-Q41215 CSJ056455 Spk D, 100ppb Spike) DOU-FG1-MSW002-O-041216 (C0056456 spk 1,10 ppb Spike) DO U-F01-MSW 002-0-041216 {C0056456 SpkJ, 100ppb Spike) D L 2 -F 0 1 -IF W 0 0 4 -0 -0 4 1 2 1 S {CQ05645?Spk C. 10 ppb Spike) DL2-F01 ~!PW004-0-041215 (C0056457Spk DH100ppb Spike) DOU-F01-MSW003-0-041216 {0)056458 Spk K, 10 ppb Spike) DOU-F01-MSW003-0-041216 (C0056458 SpkU 100 ppb Spite) Amount Spiked ..toflto). C4 Sulfonate PFBS_______________ CS Sulfonate PFHS________________ CS Suifonate PFOS Amt Found in Sample {fiiFEPwetwt, Amount Recovered Recovery fnjyg)wetwt. (% ) .... Am i Found in Sample (ng/g) wet wt. Amount Recovered np/g) wet wt Recovery i%> Am t Found in Sample (ng/g) wet wt. Amount Recovered ngigiwetwt. Recovery (%) 10 ND 100 ND 8,76 88 0.254 11.1 108 86,0 86 0,254 107 107 116 116 123 226 110 10 ND 100 ND 8,52 85 0.420 12.2 118 83,6 84 0.420 110 110 146 146 193 260 114 10 ND 100 ND 7.92 82.0 79 82 ND 10.6 106 7,92 ND 106 106 7.92 16.0 81 101 93 10 ND 7.92 79 ND 10,7 107 44.0 55.2 100 ND 83,2 83 ND 107 107 44.0 134 90 10 ND 7.88 79 ND 10,3 103 46.4 54.4 100 ND 75,6 76 ND 96.4 96 46.4 121 75 10 ND 8.44 84 ND 10.0 100 49.2 59.2 100 100 ND 83.2 83 ND 102 102 49.2 142 93 10 ND 8.00 80 ND 11,5 115 13.2 22,7 95 too ND 64.8 65 ND 107 107 13.2 121 108 10 0.206 6.92 67 1,72 10,9 92 100 0,206 67.2 67 1,72 92.8 91 NR NR NR NR NR NR 10 ND 100 ND 10 ND 100 ND 8.08 76.8 8.12 68.4 81 77 81 68 ND ND 10,4 10.4 10.4 104 6.80 95.2 95 6,80 19,4 90 105 95 NR NR 14.8 97.2 NR NR 80 90 NR NR 'Sample residue exceeds the spiking ievei significantly; therefore, an accurate recovery value canned be calculated ND = No! detected at or above Limit of Quantitation which is 0,2 ng/g (wet weight), NR * Not reported due to quality control result failures. Re-analysis results are shown in Tabie IS, Note: Since this summary table shows founded results, recovery values may vary slightly from the values In the raw data. Exygen Research Page 42 of 132 Interim Report # 10-Analysis of Fish and Clam Samples Exygen Study No.: P0001131 Table IV. Matrix Spike Recovery PFBS, PFHS and PFOS in Fish Samples (continued) Sample Description DL3- FQ1-i PW 004-0*041130 (C0056459 Spk E, 10 ppb Spike) DL3-FG14PW004-0-041130 iC00$64$9 Spk Fh100 ppb Spike) t Amount Spiked t (ng/g) C4 Sulfonate PFBS_______________ C6 Sulfonate PFHS________________ CS Sulfonate PFOS Amt Found in Sample (ng^g) wetwt Amount Recovered Recovery (ng/g) wet wt i% ) Amt Found in Sample (ng/g) wet wt. Amount Recovered Recovery ina/giwet wt. ......{% )... Amt Found in Sample (ng/g) wet wt. Amount Recovered (ng/g) wet wt. Recovery ... 1%)...... l 10 100 ND ND 7,52 66,4 75 66 ND 11.0 110 7.92 ND 118 118 7.92 17.3 94 114 106 DOU-F01-MSW0Q1 -0-041216 (CG05646G Spk a 10 ppb spike) DQU-F01 -MSW001 -0-041216 {C0056460 Spk h. 100 ppb Spike) 10 100 7.08 7.08 14,6 83.2 75 76 15.4 27.5 121 15.4 122.0 107 NR NR NR NR NR NR DO U-F01-MS WG04-0-041216 (C0056461 Spk E, 10 ppb Spike) DOU-F01-MSWOO4-0-O41216 (COOS646t Spk F, 100 ppb Spike) 10 100 4.72 4.72 10.8 72.8 61 68 21.7 21.7 29,3 104 76 82 NR NR NR NR NR NR DOU-FG1-MSWO05-0-O41216 (CG056462 spk G, 10 ppb Spike) 10 0,364 DOU-FG1-MSW005-G-041216 (C0056462 spk H, 100 ppb Spike) 100 0.364 7.08 72.0 67 72 1.34 8.52 72 1.34 87,6 86 NR NR NR NR NR NR DOU-FG1-MSFG05-0-041216 (C0056463 Spk C, 10 ppb Spike) 10 1.41 DOU-FG1-MSFOO5-0-G41216 (C005M63 Spk D. 100 ppb Spike) 100 1,41 8.60 72,0 72 71 8.40 8.40 17.6 94.0 92 86 NR NR NR NR NR NR OL3-F014PW002-0-O41130 (C005M64 Spk E, 10 ppb Spike) 10 ND D I3-F014PW 002-0-041130 (C005&464 Spk F, 100 ppb Spike) 100 ND 7,92 81.2 79 81 ND 9.04 90 12,0 ND 110 110 12.0 16,6 46 105 93 DL3-F01-iPW 003-04)41130 (CG05646S Spk G. 10 ppb Spite) 10 ND 7.96 80 ND 8.88 89 5,44 12.3 69 D L 3 -F 0 1 -IP W 0 0 3 -0 -0 4 1130 (CQ0S6465 Spk H, 100 ppb Spike) too ND 84.4 84 ND 102 102 5.44 92.0 87 DOU-F01 'MSF003-0-0412T6 (CGQ56466 Spk K, 10 ppb Spike) 10 1.72 D O U -F01 -M S F0 03-0-Q 41216 (C0056466 Spk L, 100 ppb Spike) too 1.72 9.00 89.6 73 88 6.56 6,56 14,3 77 107 100 NR NR NR NR NR NR DBC-F01-MSF005-1 -041216 (C0056467 Spk C, 10 ppb Spike) D B C - F 0 1-M S FG05-1-041216 (CO056467 Spk D, 100 ppb Spike) 10 0.220 100 0.220 8.36 83.6 81 83 1,79 11.5 97 1,79 96.8 95 NR NR NR NR NR NR DBC-FO 1-.MSF005-0-041216 (00056466 Spk K, 10 ppb Spike) DBC-F01-MSFG0S-0-O41216 (CQC564S8 Spk L 100 ppb Spike) 10 100 NO ND 8.36 80,8 84 81 1.68 11.4 97 1.68 99.6 98 NR NR NR NR NR NR 'Sample residue exceeds the spiking eve! significantly; therefore, an accurate recovery value cannot 5e calculated ND = Not detected at or above Limit of Quantitation which is 0,2 ng/g (wet weight). NR * Not reported due to quality control result failures. Re-analysts results are shewn in Table 11. Note: Since this summary table shows rounded results, recovery values may vary slightly from the values in the raw data. Exygen Research Page 43 of 132 Interim Report #10-Analysis of Fish and Clam Samples Exygen Study No.: P0001131 Table IV. Matrix Spike Recovery PFBS, PFHS and PFOS in Fish Samples (continued) Sample Description DU-F01 M$FQG2--G41216 {C05646Spk Q, 1Gppb Spite) D O U -F 0 1 -MSFO2-0-O41216 (00056469 Spk H, tOOppb Spike) D L 3 -F 0 1 -IP W 0 0 1- 0 -0 4 1130 (CO05647OSpk C. 10 ppb Spike; DL3-F01-IPWQG1 -0-041130 (CQ05647Q Spk 0 , 100 ppb Spike) DOU-F01-MSF0O2-1-D41216 (C0056471 Spk !. 10 ppb Spike) DOU-F01-MSFC02-1 -041216 (CQ56471 Spk J. 100 ppb Spite) DOU-F01-MSF0Q4-O-O41216 (CQ056472 Spk K, 10 ppb Spte) QOU-F1 "MSF004-0-041216 (00050472 Spk i, 100 ppb Spite) DOU-F01-MSF001 -0-041216 {C0056473 Spk E. 10 ppb Spike) DOU-F01-MSF001--Q41216 (C0056473 Spk F, 100 ppb Spike) DBC-F01^1SF003^041216 (C056474 Spk G, 10 ppl>Spite) DBC-FO1-MSFDG3-0-041216 (CG56474 Spk H, 100 ppb Spike) DBC-F01-MSF004-0-041216 (C0056475 spk i, 10 ppb Spike) D8C-F1-MSF0Q4-O-O41216 (C0056475 Spk J, 1(H) ppb Spite) D M C -F 0 1 -M S F 0 0 1-0-0 4 1 2 1 6 (C05S476 Spk C, 10 ppb Spite} DMC-F01-MSF0Q1-0-41216 (CO05S47S Spk Dr 100 ppb Spike) DL1-FG14FW 005-0-041216 (C0056477 Spk l 10 ppb Spike) DL1-F1- IPWQ05-0-041216 (C0056477 Spk J, 100 ppb Spike) DI2-F01-IPW003-0-041201 (COO56470 spk K, 10 ppb Spike} D L 2 -F 0 1-SP W 003-0-041201 (C005647S Spk L. 100 ppb Spike) Amount Spiked -O stS tL C4 Sulfonate PFBS Am t Found in Sample (ngfglwetwt. Amounl Recovered Recovery (fpg/g) wetwt (.%) C6 Sulfonate PFHS ____________ CS Sulfonate PFOS Amt Found Amount Am i Found in Sample Recovered Recovery in Sample (ng/g) wetwt. (bgjg) wet wt. ....0 9 . . (ng/g) wetwt. Amount Recovered (ng/g) wet wt. Recovery (%> 10 0.888 100 0.888 9.24 83.6 84 83 4.56 4,58 14.5 99 104 99 NR NR NR NR NR NR 10 ND 100 NO 7.80 78 0,246 9.80 96 9.04 82,4 82 0.246 101 1-01 9.04 17.2 82 102 93 10 0.840 100 0.840 9.20 82.0 84 81 4,68 4,68 14,5 98 103 98 NR NR NR NR NR NR 10 0.792 100 0.792 7.76 81.2 70 80 2.35 2,35 8.) 85.6 65 83 NR NR NR NR NR NR 10 2.18 100 2.18 11.1 82.8 89 81 10.9 20,8 99 10.9 104 93 NR NR NR NR NR NR 10 ND 100 ND 8.48 84.0 85 84 2,09 2.09 11,9 98 106 104 NR NR NR NR NR NR 10 NO 100 ND 7.72 75,8 77 76 3.87 3,87 12.9 98.8 90 95 NR NR NR NR NR NR 10 ND too ND 9.08 91 0.420 9.08 87 86.0 86 0.420 102 102 NR NR 10 0.253 7.40 71 0.644 9.96 93 100 0,253 72.4 72 0.644 86.8 86 68.8 68.8 NR NR 83.2 145 NR NR . 76 10 ND 100 ND 7.04 71.6 70 72 ND 9.04 SO 7,48 ND 92.0 92 7.48 13.7 84.0 62 77 'Sample residue exceeds the spiking level significantly; therefore, an accurate recovery value cannot be calculated NO = Not detected at or above Limit of Quantitation which is 0.2 ng/g (wet weight). NR = Not reported due to quality control result failures. Re-analysis results are shown in Table it Note; Since this summary table shows rounded results, recovery values may vary slightly from the values in the raw data. Exygen Research Page 44 of 132 Interim Report # 10-Analysis of Fish and Clam Samples Exygen Study No.: P0001131 Table IV. Matrix Spike Recovery PFBS?PFHS and PFOS in Fish Samples (continued) Sample Description DL3-F01-1PW005--41201 (00056479 Spk 1,10 ppb Spike) DL3-F01 -PWO5-O-041201 {C0056479 Spk J, 100 ppb Spike) Amount Spiked (ngfg) C4 Sulfonate FF8S Amt Found Amount in Sample Recovered Recovery (ng/g) wet wt. {ng/g} wet wt. ... ...... C6 Sulfonate PFHS__________ C8 Sulfonate PFOS Am t Found Amount Amt Found in Sample Recovered Recovery in Sample (ngig) wet wt. {ng/g) wet wt {%) (ngtg) wet wt. Amount Recovered ing/g) wet wt. Recovery <%) 10 ND 8.16 82 ND 8.96 90 4.88 14,0 91 100 ND 84.8 85 ND 97.2 97 4.88 94,4 90 Average: Standard Deviation: 79 13 Average: Standard Deviation: $4 12 'Sample residue exceeds the spiking level significantly; therefore, an accurate recovery vaiue cannot he calculated ND = Not detected at or above Limit of Quantitation which is 0,2 ng/g (wet weight), NR = Not reported due to quality control result failures. Re-analysis results are shown In Table 1!. Note: Since this summary table shows rounded results, recovery values may vary slightly horn the values in the raw data. Average; Standard Deviation: 85 20 Exygen Research Page 45 of 132 Interim Report # 10-Analysis of Fish and Clam Samples Exygen Study No.: P0001131 Table V. Matrix Spike Recovery of PFOS in Re-extracted Fish Samples S am pie D escription D B C - F 0 1-1P W 0 0 1 -0 -0 4 1 0 2 1 (C0048151 Spk C, 1000 ppb) D B C - F 0 1-t P W 0 0 2 -0 - 0 4 1021 (C048152 Spk D, 5000 ppb) D B C -F 01-P W 03-0-041021 (C0G48153 Spk E, 1000 ppb) D O U -F01 -iP F O 0 1-0 -04 1 0 21 (C004S154 Spk F, 1000 ppb) D O U -F 0 1-IP F 0 0 2 -0 -0 4 1021 (C0048155 Spk G, 1000 ppb) D O U -F 1-IP F Q 5 -Q - 4 1021 (CQ48158 Spk H, 1000 ppb) D O U -F 0 1-P W 0 0 2 -0 -0 4 1021 (C0048160 Spk 1,1000 ppb) D BC -F01-M SW 01-0-0411G 4 (C049450 Spk J, 10,000 ppb) D 8C -F 1-MSW G2-0-G41104 (CGQ49451 Spk K. 10,000 ppb) D B C -F 01 -M S W 0 03-0 -0 41104 (CQ049452 Spk L, 10,000 ppb) D B C -F 01-M S W 0 0 4-0 -0 4 1104 (C04S453 Spk M, 5000 ppb) D 8 C -F 0 1-M S W 0 0 5 -0 -0 4 1104 (C0049454 Spk N, 10,000 ppb) D B C -F 0 1-M S F O 0 143-041104 (C0049455 Spk C, 10,000 ppb) D B C -F 1-M S FQ 2- - 41104 {C004S456 Spk D, 10,000 ppb) D 8 C -F 0 1-IP F 0 0 5 -0 -0 4 1 104 (C0049458 Spk E, 1000 ppb) D B C -F 0 1-IPW QQ4~0~041104 (CQ049458 Spk F, 1000 ppb) D B C -F 0 1-i PW 0 0 5 -0 -0 4 1104 (0)049460 Spk G, 50 ppp) D O U - F 0 1-1P W 0 0 3 -0 -0 4 1 1 0 4 (C04S461 Spk H, 1000 ppb) D O U -F 01'!P W 004-G -041104 (C0049462 Spk i, 1000 ppb) D O U -F01 -1PW 0Q5-0-G41104 (C0049463 Spk j , 5000 ppb) D U -F 0 1-M S W 001 -0 -0 4 1 1 0 4 (CG4S4S5 Spk K, 1000 ppb) D L1-F01-M S W 002-0-041104 (C049466 Spk C, 1000 ppb) D L 1-F 0 1-M S W 0 3-0 - 4 1104 (C049467 Spk D, 1000 ppb) D L 1-F 0 1-M S W 0 0 5 -0 -0 4 1104 (03049469 Spk E, 1000 ppb) Am ount S p ike d (rtg/g) 1000 5000 1000 1000 1000 1000 1000 1G00G 10000 10000 5000 10000 10000 10000 1000 1000 5000 1000 1000 5000 1000 1000 1000 1000 C8 Sulfonate PFOS Am t Found A m ount in S a m p le {ng/g) w et w t R ecovered (n g /g ) w e t wt. R ecovery (% ) 2080 2860 78 3330 8680 107 1660 3160 150 596 1760 116 636 1740 110 544 1650 111 1210 2610 140 9480 23200 137 7520 16600 91 12600 23200 106 3440 8520 102 5240 15200 100 5240 14400 92 4760 15200 104 608 1:600 99 1230 2590 136 2620 6520 78 2270 2640 37* 1740 2740 100 2130 7600 109 968 1860 89 708 1780 107 696 1930 123 604 1680 108 ` H ot reported due to quality c ontrol resuit failures. Values are not included in the sum m ary statistics. Exygen Research Page 46 of 132 Interim Report #10-Analysis of Fish and Clam Samples Exygen Study No.: P0001131 Table V. Matrix Spike Recovery of PFOS in Re-extracted Fish Samples S am ple D escription (continued) C8 Sulfonate PFOS Am ount Am t Found Am ount S p ik e d (ng/g) in S a m ple (np/g) w et w t R e covered fnq/cj) w et wt. R e co ve ry (% } D L 1-F 01-M S F 002-0-0411Q 4 (C0G49471 Spk f , 1000 ppb} 1000 568 1690 ' 112 D L1-F O 1-M S F 0 3-O -O 4 1104 (CG049472 Spk G, 1000 ppb) 1000 432 1670 124 D L1-F01-M SF005-0-041104 (C0049474 Spk H, 1000 ppb) 1000 740 1830 109 DM C -F 0 1-M S W 0 G 5-0-041105 (C0049496 Spk I. 1000 ppb) 1000 560 1780 122 D M C -F01-M S F 002-0-041105 (C0049499 Spk J, 1000 ppb) 1000 620 1500 88 DOU-F01-M SW 0O2-G-G41216 (C0056456 Spk K, 10,000 ppb) 10000 28200 32700 45 DOU-F01-MSW G3-O-O41216 (C0056458 Spk L, 5000 ppb) 5000 4040 10200 123 D O U-F01-M SW 001-0-041216 (C0056460 Spk C, 10000 ppb) 10000 7600 25000 174* DOU-F01 -M S W 004-0-041216 (C0056461 Spk D, 5000 ppb) 5000 5120 9880 95 DO U -F01-M SW 005-0- 41216 (C0056462 Spk E, 1000 ppb) 1000 4% 1780 128 D O U -F01-M SF005-0-041216 (CG056463 Spk F, 1000 ppb) 1000 720 1820 110 DO U-F01-M SF003-0-041216 (C0056466 Spk G, 1000 ppb) 1000 848 1720 87 D B C -F 0 1-M S F0G 5-1-041216 (C0056467 Spk H, 10000 ppb) 10000 9320 32400 230* D B C -F 0 1-M SFG05-0-G 41216 (CQ056468 Spk I, 10000 ppb) 10000 8960 24100 151 DOU-F01-M SF0G2-G-O41216 {CQ05646S Spk j , 1000 ppb) 1000 640 1720 108 DOU-F01-M SF0G2-1 -041216 (COO56471 Spk K, 1000 ppb) 1000 692 1950 126 D O -F 01-MSF0G4-G-G41216 (C0056472 Spk l , 1000 ppb) 1000 1820 2700 88 D O U -F01 -M S FO 01-0-041216 (COO56473 Spk C, 5000 ppb) 5000 2070 7960 118 DBC-FG 1-M SF003-0-041216 (C0056474 Spk D, 10000 ppb) 10000 4640 15500 109 D B C -F 0 1-M S F0G 4-0-041216 (CC056475 Spk , 10000 ppb) 10000 9280 24500 152 DM C -F01 -M S FO 01-0-041216 (COO56476 Spk F, 1000 ppb) 1000 840 2040 120 D B C -F 0 1 -IP F 0 0 1-0-041021 (CG048148 Spk G, 1000 ppb) 1000 384 1350 97 D 8C -F01 -i P F 0 03-0-041021 (C0O481SO Spk H, 1000 ppb) 1000 2S3 1340 105 D O U -F 0 1 -1 P F 0 0 3 -0 -0 4 1021 (C0O48156 Spk i, 1000 ppb) j 1000 359 1400 104 *Not reported due to quality control result failures- Values are not included in the sum mary statistics - Exygen Research Page 47 of 132 Interim Report #10-Analysis of Fish and Clam Samples Exygen Study No. : P0001131 Table V. Matrix Spike Recovery of PFOS in Re-extracted Fish Samples (continued) Sample Description DOU-FO1-IPF04-0-041021 (C048157 Spk J. 1000 ppb) DOU-F01-IPW001-0-041021 (00048155 Spk K, 1000 ppb) DL1-F01 -MSF004-0-041104 (C0Q4S473 Spk L. 1000 ppb) Amount Spiked (ng/g) 1000 1000 1000 C8 Sulfonate PFOS Amt Found Amount in Sample Recovered (ng/g) wet wt. {ng/g) wet wt. Recovery <%> 424 1520 110 387 1480 109 348 1400 105 Average; Standard Deviation; 109 20 Exygen Research Page 48 of 132 Interim Report #10-Analysis of Fish and Clam Samples Exygen Study No.: P0001131 Table VI. Matrix Spike Recovery PFBS, PFHS and PFOS in Clam Samples Sam pie Description DL341-CFWQ1-04341216 {CG564SGSpk C, 10 ppo DI3-I1 -CFWO01-Q-41216 (000564805^0, 100 ppb) DMC4Q1-CFWQQ1*(M)41216 (C00564S1 $pk Er 10 ppb) DMO1G1-CFW001-0-041216 (00056461 Spkf=. 100 ppb) DL2401-CFW001 -0-041216 (C0Q564&2 Spit G, W ppb) DL2-01-CFW00143-041216 {C56462Spk H, 100 pp^> DBG-01-CFW001-0-041216 (C056463 Spk f iGppb) DBC-I01-CFW001-0-0412t6 (C0056403 Spk Jr lOOppb) DL1401-CFW00143-41215 {C05&44 Spk K 10 ppb) DL1-101-CFWO01*04)41215 (00056404 Spk Lt 100ppD) DOU-t01-CFWQQ1-0-041216 (CQ056485 Spy M. 10 ppb) DOU-iO 1-CFWO01-0-041216 (C00564S5 Spy N. 100 ppb) Amount Spiked (ng/g) C4 Sulfonate PF8S_________________ C6 Sulfonate PFHS________________ C8 Stifo naie PFOS Amt Found Amount Amt Found Amount Amt Found Amount in Sample {flQ/iriwSlwi. Recovered Recovery in Sam pie in#i)wet wt. (%) (vg/g)w$<M. Recovered (ng/g) we! wt. Recovery in Sample f%) (npig) wet w*. Recovered Recovery (nFg)w&i wi. f%) 10 ND 5,96 60 N 6.16 62 1.82 6.32 45 100 ND 82,4 82 ND 95.6 96 1.82 85.2 83 10 ND too ND 9.04 90 ND 82.8 83 ND 9.44 98.8 94 6.16 99 6,16 11.3 51 92.8 87 10 N D too ND 7.28 73 ND 83.6 84 ND 7.84 78 1.81 101 101 1.81 7.52 57 88.4 87 10 ND 100 ND 10 ND 100 ND 10.3 103 ND 84.4 84 ND 8.96 90 ND 85.2 85 ND 10.8 97.2 9.32 103 108 4.48 97 4.48 93 8.80 103 8.80 13.0 85 86.4 82 15,0 62 96.8 88 10 ND 100 ND 8,48 85 0.202 8.32 81 11.4 17.2 58 84.4 84 0.202 98.0 98 11 4 98.0 87 Average: 84 Standard Deviation: 10 Average: Standard Deviation; ND = Not detected at or above Limit of Quantitation which is 0.2 ng/g (wei weight), Note: Since Sits summary table shows rounded results, recovery values may vary sitghBy from (he values in the raw data. 3 13 Average; 73 Standard Deviation: 16 Exygen Research Page 49 of 132 Interim Report #10-Analysis of Fish and Clam Samples Exygen Study No.: P0001131 r 1 Vjr U r v l L 3 Exygen Research Page 50 of 132 Interim Report #10~Analysis of Fish and Ciam Samples Exygen Study No,: PlDl Figure 1. Typical Calibration Curve for PFBS in Methanol G8165E Fbh.rdb (PFS); "L in e a r" Regression (" 1 />? w e ig htin g ); y = 1.31 e+4 x + 3.07e+Q03 <r = 0.98913) Ares, counts Exygen Research Page 51 of 132 Interim Report # 10-Analysis of Fish and Clam Samples Exygen Study No.: P01131 Figure 2. Nun-Extracted Standards of PFBS in Methanol, 0.0005 pg/mL and 0.001 pg/mL, Respectively I SS7279 PFBS (Standard) 29SUVW .9ama -samp} 1 o f 39 fro m O6?60SEmW Area; 30O3 counts fie ig ftt: 3 5 f. cps RT: 0.572# 0.51 Area: 16857 eaunti Height: 781. cps RT: 0.515 min 0.52 Intensity, cps Intensity, cps Exygen Research Page 52 of 132 Interim Report # 10-Analysis of Fish and Clam Samples Exygen Study No. : P0001131 Figure 3. PFBS in Control Fish Blank, 0.00625 pg/mL Fortified Control Fish Spk A, and 0.025 pg/mL Fortified Control Fish Spk B, Respectively Raagettt C o n tro l - PFBS (Unknown) 299.9/99,9 am u -sam ple 9 o f 39 fro m &61$95E,wtff (peak no t fo o tid } 8 9 10 11 12 13 14 15 16 17 Time, min I R eagent Spk A - PFBS (Q C ) 2 98 .0/9 9.0 amu - sam ple 10 of 38 ite m 0 8 1 6 0 5 E.w ifi Area: 7 4 4 7 4 counts H e ig h t 1160. ops RT: 4 .42 min 1 2 3 4 5 6 7 8 8 10 11 12 13 14 15 16 17 Tim e, min R eagent Spk B - PFBS (Q C )2 69 .0/8 8.0 amu - sample 11 ef 38 from 0618G6.sylff A fe a : 335 32 0 counts H eight: 18900. ops RT: 0 .527 m in 0.53 Exygen Research Page 53 of 132 Interim Report #10-Analysis of Fish and Clam Samples Exygen Study No.: P01131 Figure 4. Chromatogram Representing a Fish Sample Analyzed for PFBS (Exygen ID: C0049478, Data Set:06I6O5E) C049479 PFBS (UiHiHQwtt} 299.m 9.0a/tts -am pin 20 o f 38 fro m iW U S E .w tff " Arna: 4127 counts H nigkt: 69.3 cp s RT: 0.S4min intensity, cps Exygen Research Page 54 of 132 Interim Report # 10-Analysis of Fish and Clam Samples Exygen Study No.: P1131 Figure 5. Typical Calibration Curve for PFHS in Methanol 051S5E Fish,fdb (PFHS): "Lineai1*Recession {"1 t x11weighting); y = 2,8Se+004x + 3 ,84e+0G3 (r = 0,9981) Area, counts Exygen Research Page 55 of 132 Intensity, cps Interim Report # 10-Analysis of Fish and Clam Samples Exygen Study No.: P001131 Figure 6, Non-Extracted Standards ofPFH S in Methanol, 0.0005 pg/mL and 0.001 pg/mL, Respectively SS7279 - PFtiS s ta n d a rd } 3S&1W .0 am -sam ple 1 o f 39 fro m tWSQSE.wtff Area: W 3S co asts H eight; 749. cps ftT : 19.3 m in 10.32 Tim, min SS727S - PFHS (Standard) 388.0/30.0 amu sampie 2 o f 38 from 061805E,cuiff Area: 33178 counts H eight: 13SO, cps RT: 10.3 min 10,33 12001000800800- 400- 2000J-- 1----- 1----- 1----- 1-- --- 1----- --- --- !----"r 1 23 45 67 8 Time, min Intensity, cps Exygen Research Page 56 of 132 Interim Report #10-Analysis of Fish and Clam Samples Exygen Study No.: P0001131 TFr*migure 7urn. PFHS in Contaroli HFani*shi 'Barli anki , 0a .0\0\625p* pg//mL Forjtigf**iedi Control Fish Spk A, and 0.025 pg/inL Fortified Control Fish Spk B, Respectively | Reagent C o n tro l - PFHS (Unkttewtt} 3 9 9 .m M a m a -sampte 9 o f 38 fro m Q616$5.wffi (peak Not found) | R e ag e nt Spk A - PFHS ( 0 )3 9 8 ,0 /8 0 ,0 am u - sam ple 10 Of 3 8 from Q$18G5E.wiff Area: 186711 counts H e ig ht: 12000. ops RT: 1 0 3 min i/> g - 1.00 e4 10.32 2<0 5000.00 8 9 10 Time, min | Reagent Spk B - PFHS (0)399.0/80.0 im u - sam ple 11 of 38 from 061805,tariff Area: 721871 counts Height: 30100. cps RT: 10.4 min Exygen Research Page 57 of 132 Interim Report #10~Analysis of Fish and Clam Samples Exygen Study No.: P0001131 Figure 8. Chromatogram Representing a Fish Sample Analyzed for PFHS (Exygen ID: C0049478, Data Set: 061605E) I C0049479 PFHS (Unknown) 399.0/80.0 nmn sample 20 o f 38 fro m 961605.wtff Aron; 21019 c o m t t H eight; 1130. c p t RT: 10,3 s tilt Intensify, cps Exygen Research Page 58 of 132 Interim Report #10-Analysis of Fish and Clam Samples Exygen Study No.: P0001131 Figure 9. Typical Calibration Curve for PFS in Methanol 061S05E Fish.rdb (PFOS): "Lineat" Regression ("1 /x " weighting): y m 1 15e+004 x + 3.36e+GQ3 ft * 0.9998} Area, counis Exygen Research Page 59 of 132 Interim Report #10-Analysis of Fish and Clam Samples Exygen Study No.: P0001131 Figure 10. Non-Extracted Standards of PFOS in Methanol, 0.0005 fig/mL and 0.001 (xg/mL, Respectively I SS7279 -PFOS {Standard) 4m 8 iff -sampte 1 o f M from m iM SE .w ff Area: 8368 counts fieigkt: 432. Cps ftT: 72.5 mia intensify, cps Afea: 1487 counts Haight: BOO. ops RT: 12,5 min 12,46 Intensify, cps Exygen Research Page 60 of 132 Intensity, ops Interim Report #10-Analysis of Fish and Clam Samples Exygen Study No.: P0001131 Figure 11. PFOS in Control Fish Blank, 0.00625 pg/mL Fortified Control Fish Spk A, and 0.025 pg/mL Fortified Control Fish Spk B, Respectively I ife s g s ift C ontrol * PFOS (Unknown) 499.0/t&.S amn -sam ple 9 o f 38 fro m t}616d5,wiff A rea: Z2SS counts H eight; S0.4 eps RT: 1Z$ a tilt 5.44 h J S U TM. .. s .. 9.20 0.87 1 2 3 4 5 6 7 8 9 10 Tima, min Reagent Spk A - PFOS (QC) 49S.OiSO.O amu - sample 10 of 38 from G01SO5E.wiff Area: 71990 counts Height: 4640, ops RT: 12.5 min | Reagent Spk S - PFOS (QC) 488,0/80.0 amu sample 11 of 38 from G810G5E.<ftriff Area: 287141 counts Height: 169. ops RT: 12.5 min 12.49 intensify', cps fntensity.eps Exvgen Research Page 61 of 132 Interim Report # 10-Analysis of Fish and Clam Samples Exygen Study No.: P0001131 Figure 12. Chromatogram Representing a Fish Sample Analyzed for PFOS (Exygen ID: C0049478, Data Set:0616O5E) intensify, cps I CQ04M79 PFO S Jttkmown} - sampfn 20 o f 38 fro m 001005E.wiff u ta : 628966co asts H ogitt; 35M 0. cps RT: 1 2 5 m in 1,3e5 - 1,2e5 - 1.1*5 1.0*5 - 9.0*48.0*4- 7,0*48.0*4- 5.0*4- 4,0*4 - 3,0*4 - 2.0*4- 1,0e4 - 0.0 ---------.---- -----1--- -----1---- ----- 1---- -----1--- -----,--- -- r 12 345 67 Time, min 12.47 Exygen Research Page 62 of 132 Interim Report #10-Analysi$ of Fish and Clam Samples Exygen Study No.: P0001131 APPENDIX A Study Protocol P0001131 (Exygen Study No. P0001131) with Analytical Method, Protocol Amendments 1 and 2 and Deviations 2 and 3 Exygen Research Page 63 of 132