Document 7RN7gjrQ5EdXpMGxENaZ8GbD6

INTERIM REPORT #22 - Analysis of Soil Samples STUDY TITLE Analysis of Perfluorooctanoic Acid (PFOA) in Water, Soil, Sediment, Fish, Clams, Vegetation, Small Mammal Liver and Small Mammal Serum Using LC/MS/MS for the 3M Decatur Monitoring Program DATA REQUIREMENTS EPA TSCA Good Laboratory Practice Standards 40 CFR 792 STUDY DIRECTOR Jaisimha Kesari P.E., DEE Weston Solutions, Inc. 1400 Weston Way West Chester, PA 19380 Phone: 610-701-3761 INTERIM REPORT COMPLETION DATE September 22,2006 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: P0000760 Exygen Study Number: P0000760 Total Pages: 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT Exygen Study Number P0000760, entitled "Analysis of Perfluorooctanoic Acid (PFOA) in Water, Soil, Sediment, Fish, Clams, Vegetation, Small Mammal Liver and Small Mammal Serum Using LC/MS/MS for the 3M Decatur Monitoring Program," conducted for 3M Company, is being performed in compliance with EPA TSCA Good Laboratory Practice Standards 40 CFR 792 by Exygen Research. Exygen Research Jaisimha E Study Director Weston Solutions, Inc. Michael A. S Sponsor Repi 3M Company Exygen Research Page 2 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 QUALITY ASSURANCE STATEMENT Exygen Research's Quality Assurance Unit reviewed Exygen Study Number P0000760, entitled, "Analysis of Perfluorooctanoic Acid (PFOA) in Water, Soil, Sediment, Fish, Clams, Vegetation, Small Mammal Liver and Small Mammal Serum Using LC/MS/MS for the 3M Decatur Monitoring Program". All reviewed phases1 were inspected for conduct according to Exygen Research's Standard Operating Procedures, the Study Protocol, and all applicable Good Laboratory Practice Standards. All findings were reported to the Exygen Principal Investigator and Management and to the Study Director. Phase Date Inspected Date Reported to Date Reported to Principal Exygen Date Reported to Investieator Management Studv Director 32. Raw Data Review and Interim Analytical Report Review 06/06-09/06 06/29/06 06/30/06 07/07/06 33. Interim Analytical Report Review 34. Raw Data Review and Interim Analytical Report Review 06/14/06 09/20-21/06 06/29/06 09/22/06 06/30/06 09/22/06 07/07/06 09/22/06 d ld l* )_______________ LydiaMaffer I J /J Technical Lead, Quality Assurance Unit Date ________ 'Note: All in-lab inspections will be documented in the QA statement for the final analytical report at the conclusion of the study. This QA statement involves only the review of the interim report and associated raw data. Exygen Research Page 3 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 CERTIFICATION OF AUTHENTICITY This interim report, for Exygen Study Number P0000760, is a true and complete representation of the raw data. Submitted by: Exygen Research 3058 Research Drive State College, PA 16801 (814) 272-1039 Principal Investigator, Exygen: Date Exygen Research Facility Management: y z-J O * - 7t Date Study Director, Weston Solutions, Inc. Jaisimha KesaK P.E., DEE Weston Solutions, Inc. Sponsor Representative, 3M Company: Wn Jj / Michael A. Smzftoro Director of Regulatory Affairs Exygen Research Da/e / Page 4 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 STUDY IDENTIFICATION Analysis of Perfluorooctanoic Acid (PFOA) in Water, Soil, Sediment, Fish, Clams, Vegetation, Small Mammal Liver and Small Mammal Serum Using LC/MS/MS for the 3M Decatur Monitoring Program PROTOCOL NUMBER: P0000760 EXYGEN STUDY NUMBER: P0000760 TYPE OF STUDY: Residue SAMPLE MATRIX: Soil and Water TEST SUBSTANCE: Perfluorooctanoic acid (PFOA) SPONSOR: 3M Company 3M Building 0236-01-B-10 St. Paul, MN 55144 STUDY DIRECTOR: Jaisimha Kesari P.E., DEE Weston Solutions, Inc. 1400 Weston Way West Chester, PA 19380 STUDY MONITOR: Michael A. Santoro 3M Company 3M Building 0236-01-B-10 St. Paul, MN 55144 PERFORMING LABORATORY: Exygen Research 3058 Research Drive State College, PA 16801 ANALYTICAL PHASE TIMETABLE: Study Initiation Date: 11/05/04 Interim Analytical Start Date: 04/15/06 Interim Analytical Termination Date:07/24/06 Interim Report Completion Date: 09/22/06 Exygen Research Page 5 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 PROJECT PERSONNEL The Study Director for this project is Jaisimha Kesari at Weston Solutions, Inc. The following personnel from Exygen Research were associated with various phases of this interim portion of the study: Name John Flaherty Karen Risha Eric Edwards Christine Edwards Mark Ammerman Amy Sheehan Mindy Cressley Brittany Kravets Krista Gallant Scott Crain Frances Crespi Sharareh Zolghadr Larissa Hoover Brian McAllister Title Vice President Scientist Sample Custodian Technician Sample Custodian Associate Scientist Technician Technician Technician Technician Technician Technician Technician Sample Custodian Exygen Research Page 6 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 TABLE OF CONTENTS Page TITLE PAGE....................................................................................................................... 1 GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT..............................2 QUALITY ASSURANCE STATEMENT.......................................................................... 3 CERTIFICATION OF AUTHENTICITY...........................................................................4 STUDY IDENTIFICATION................................................................................................ 5 PROJECT PERSONNEL.....................................................................................................6 TABLE OF CONTENTS.....................................................................................................7 LIST OF TABLES............................................................................................................... 8 LIST OF FIGURES..............................................................................................................9 LIST OF APPENDICES.................................................................................................... 10 1.0 SUMMARY................................................................................................................ 11 2.0 OBJECTIVE............................................................................................................... 12 3.0 INTRODUCTION....................................................................................................... 12 4.0 ANALYTICAL TEST SAMPLES.............................................................................. 12 5.0 REFERENCE MATERIAL........................................................................................ 13 6.0 DESCRIPTION OF ANALYTICAL METHOD........................................................ 15 6.1 Extraction Procedure For Soil.................................................................................. 15 6.2 Percent Solids Procedure For Soil............................................................................ 15 6.3 Extraction Procedure for Water................................................................................ 15 6.4 Preparation of Standards and Fortification Solutions............................................... 16 6.5 Chromatography....................................................................................................... 17 6.6 Instrument Sensitivity............................................................................................... 17 6.7 Description of LC/MS/MS Instrument and Operating Conditions...........................17 6.8 Quantitation and Example Calculation..................................................................... 18 7.0 EXPERIMENTAL DESIGN......................................................................................20 8.0 RESULTS...................................................................................................................21 9.0 CONCLUSIONS.........................................................................................................21 10.0 RETENTION OF DATA AND SAMPLES................ 22 Exygen Research Page 7 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table I. LIST OF TABLES Page Summary of PFOA in Soil Samples................................................................24 Table H Summary of PFOA in Re-extracted Soil Samples...........................................32 Table HI. Summary of PFOA in Associated Rinse Blanks and Trip Blanks................. 33 Table IV. Matrix Spike Recovery of PFOA in Soil Samples......................................... 34 Table V. Matrix Spike Recovery of PFOA in Re-extracted Soil Samples.................... 46 Table VI. Matrix Spike Recovery of PFOA in Associated Trip Blanks......................... 48 Table VH. Surrogate Spike Recovery of 13C PFOA in Soil Samples............................. 49 Table VUI. Surrogate Spike Recovery of 13C PFOA in Re-extracted Soil Samples....... 61 Table IX. Surrogate Spike Recovery of 13C PFOA in Associated Rinse Blanks and Trip Blanks.................................................................................................... 63 Table X. Total Percent Solids in Soil Samples............................................................. 64 Exygen Research Page 8 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Figure 1. LIST OF FIGURES Page Typical Calibration Curve for PFOA in Methanol........................................ 70 Figure 2. Extracted Standards of PFOA in Methanol, 25 ng/mL and 50 ng/mL, Respectively.................................................................................................... 71 Figure 3. PFOA in Methanol Reagent Blank, 50 ng/L Fortified Reagent Spk A, and 500 ng/L Fortified Reagent Spk B, Respectively................................... 72 Figure 4. Chromatogram Representing a Soil Sample Analyzed for PFOA (Exygen ED: C0156344, Data Set: 041206AR)..............................................73 Figure 5. Typical Calibration Curve for 13C PFOA in Methanol.................................. 74 Figure 6. Extracted Standards of 13C PFOA in Methanol, 25 ng/mL and 50 ng/mL, Respectively.................................................................................................... 75 Figure 7. Figure 8. 13C PFOA in Methanol Reagent Blank, 50 ng/L Fortified Reagent Spk A, and 500 ng/L Fortified Reagent Spk B, Respectively.....................................76 Chromatogram Representing a Soil Sample Analyzed for 11C PFOA (Exygen ID: C0156363, Data Set: 041306A).................................................77 Exygen Research Page 9 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 LIST OF APPENDICES Paee Appendix A Study Protocol P0000760 (Exygen Study No. P0000760) with Analytical Methods and Protocol Amendments....................................... 78 Exygen Research Page 10 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 1.0 SUMMARY Exygen Research extracted and analyzed soil samples for the determination of perfluorooctanoic acid (PFOA) according to Exygen Method V0001781 and water samples for the determination of perfluorooctanoic acid (PFOA) according to Exygen Method V0001780 (Appendix A). Thirty-three of the analyzed soil samples were not reported (NR) due to quality control failure. Eight of the thirty-three samples failed because the PFOA spikes were too low relative to the endogenous level in the sample to allow assessment of matrix interference. Twenty-five of the thirty-three samples failed for unknown reasons. Additional analytical work was performed in an attempt to provide quantitative results for these thirty-three samples. Quantitative PFOA results were obtained for all samples except for nine soil samples that are not reported due to quality control failures. The limit of quantitation for PFOA in soil was 0.2 ng/g. The LOQ provided is assuming wet weight values and the results for the actual samples are reported based on dry weight. The limit of quantitation for PFOA in water was 25 ng/L. Analytical results and assessed accuracies for the analysis of PFOA in soil samples based on dry weight are summarized in Table I. Analytical results and assessed accuracies for the analysis of PFOA in re-extracted soil samples based on dry weight are summarized in Table II. Analytical results and assessed accuracies for the analysis of PFOA in water samples are summarized in Table III. Field matrix spikes that were in the appropriate concentration range (primary concentration must be less than or equal to three times the spiking level) were used to calculate the assessed accuracies. When the field matrix spikes were not available or failed to meet recovery criteria, the laboratory spikes were used. Fortification recoveries for the analysis of PFOA in soil samples based on wet weight are summarized in Table IV. The overall average percent recovery standard deviation for PFOA in soil samples based on wet weight was 80 19%. Fortification recoveries for the analysis of PFOA in re-extracted soil samples based on wet weight are summarized in Table V. The overall average percent recovery standard deviation for PFOA in re extracted soil samples based on wet weight was 74 18%. Fortification recoveries for the analysis of PFOA in water samples are summarized in Table VI. The overall average percent recovery standard deviation for PFOA in water samples was 92 7%. Fortification recoveries for 13C PFOA in soil samples based on wet weight are summarized in Table VII. The average percent recovery standard deviation for 13C PFOA in soil samples based on wet weight was 91 32%. Fortification recoveries for C PFOA in re-extracted soil samples based on wet weight are summarized in Table VIII. The average percent recovery standard deviation for C PFOA m re-extracted soil Exygen Research Page 11 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 11 , samples based on wet weight was 48 30%. Fortification recoveries for C PFOA in water samples are summarized in Table IX. The average percent recovery standard deviation for 13C PFOA in water samples was 107 14%. The total percent solids for soil samples are detailed in Table X. 2.0 OBJECTIVE The objective of the analytical part of this study was to determine levels of perfluorooctanoic acid (PFOA) in soil and water according to Protocol P0000760 (Appendix A). 3.0 INTRODUCTION This report details the results of the analysis for the determination of PFOA in soil using the analytical method entitled, "V0001781: Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS" and for the determination of PFOA in water using the analytical method entitled, "V0001780: Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS." The study was initiated on November 5, 2004, when the study director signed protocol number P0000760. The analytical start date for this interim report was April 15, 2006 and the analytical termination date for this interim report was July 24,2006. 4.0 ANALYTICAL TEST SAMPLES Five water samples, representing one trip blank and its associated spikes and two rinse blanks (RB), and forty-seven soil samples (Exygen ID C0156344 - C0156395) were received on wet ice on March 2, 2006 from Tim Frinak at Weston Solutions, Inc. Six water samples, representing one trip blank and its associated spikes and three rinse blanks (RB), and twenty-three soil samples (Exygen ID CO159443 - CO159471) were received on wet ice on March 10, 2006 from Tim Frinak at Weston Solutions, Inc. Four water samples, representing one trip blank and its associated spikes and one rinse blank (RB), and thirty-four soil samples (Exygen ID CO160867 - CO160904) were received on wet ice on March 15, 2006 from Tim Frinak at Weston Solutions, Inc. Four water samples, representing one trip blank and its associated spikes and one rinse blank (RB), and thirteen soil samples (Exygen ID CO164222 - CO164260) were received on wet ice on March 24, 2006 from Tim Frinak at Weston Solutions, Inc. Please note that rinse blank (RB) samples do not have associated field spikes or laboratory spikes. The samples were logged in by Exygen personnel and placed in refrigerated storage. Exygen Research Page 12 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 The soil samples collected represent both soil boring samples and surface soil samples. Sample ID coding for the soil samples consisted of five sections or strings in the following format: Dxxx-xx-xxxxxx-x(x)-xxxx. The first string of code, for the soil boring samples, contains DAL, which represents Decatur, Alabama. For the surface soil samples, the first string begins with D for Decatur, Alabama and the xxx defines the sampling location within the study area where: DF05 = Field 5 DF11 = Field 11 DF13= Field 13 DF10 = Field 10 DF12 = Field 12 The second string defines the type of sample where: SB = soil boring sample SS = surface soil sample The third string for the soil boring samples defines the soil boring identification. For the surface soil sample, the third string defines the location within the sampling area. The fourth string more specifically defines the sample type where: 0 = primary field sample RB = equipment rinseate blank sample DB = field duplicate sample The fifth string defines the collection depth of the soil samples where: 0000 = 0 to 3 inches 0005 = 6 inches to 1 foot 0050 = 5 to 5.5 feet 0003 = 3 to 6 inches 0010 = 1 to 1.5 feet 0100 = 10 to 11 feet Sample login and chain of custody information is located in the raw data package associated with this interim report. Storage records will be kept at Exygen Research. 5.0 REFERENCE MATERIAL The analytical standard, PFOA (PFOA SP0005444), was purchased from Oakwood Products, Inc. and was received at Exygen on March 17, 2005. The analytical standard, PFOA (PFOA SP0007768), was received at Exygen on June 13, 2006 from the 3M Company. The surrogate spiking standard, 13C labeled perfluorooctanoic acid (13C PFOA SP0004184), was received at Exygen on April 15, 2004 from the 3M Company. The surrogate spiking standard, 13C labeled perfluorooctanoic acid (13C PFOA SP0006528), was received at Exygen on October 13, 2005 from the 3M Company. The surrogate Exygen Research Page 13 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 spiking standard, 13C labeled perfluorooctanoic acid (13C PFOA SP0007767), was received at Exygen on June 13,2006 from the 3M Company. The available information for the reference materials is listed below. PFOA was stored ambient and 13C PFOA was stored frozen. Comnound PFOA PFOA 13C PFOA 13C PFOA 13C PFOA Exveen Inventory No. SP0005444 SP0007768 SP0006528 SP0004184 SP0007767 Lot# 203 HU14401DU 3507-195 3507-195 3507-277 Puritv (% ) 99 96.7 97 97 97.6 Expiration Date 03/31/07 12/15/06 03/29/09 03/29/09 07/15/10 The molecular structure of PFOA is given below: PFOA Chemical Name: Perfluorooctanoic acid Molecular Weight: 414 Transitions Monitored: 413 - 369 (for quantification) and 413 -->219 (for confirmation) Structure: 13C PFOA Chemical Name: 1,2-13C perfluorooctanoic acid Molecular Weight: 416 Transition Monitored: 415 -> 370 Structure: F FF F F FF FO F F OH Exygen Research Page 14 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 6.0 DESCRIPTION OF ANALYTICAL METHOD The analytical methods "V0001781: Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS" and "V0001780: Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS" were used for this study. 6.1 Extraction Procedure For Soil Before the samples were weighed for the extraction, they were placed into a new, clean Ziploc bag and mixed thoroughly. The samples were then transferred back to the sampling container. A 5-gram portion of soil was weighed into a fifty-milliliter centrifuge tube for the extraction. After fortification of appropriate samples, 5 mL of methanol was added to the samples. The samples were allowed to shake on a wrist action shaker for -15 minutes and were then sonicated in an ultrasonic bath for ~15 minutes. The volume was taken to 40 mL with water and the samples were then centrifuged for -10 minutes at -3000 rpm. The supernatant was then loaded onto a Cjg SPE cartridge conditioned with 10 mL of methanol and 5 mL of water. The eluate was discarded. Approximately five milliliters of methanol was added to the cartridge. Five milliliters of eluate was collected into a graduated 15 mL polypropylene centrifuge tube. Each sample was analyzed by LC/MS/MS electrospray. 6.2 Percent Solids Procedure For Soil Percent solids were determined using the procedure indicated in Exygen method V0000427. Approximately 20 grams of sample was weighed into a pan. The weight of the sample plus the pan was recorded. The samples were then dried in an oven overnight at 104 2 C. Then the samples were transferred to a dessicator and allowed to cool for -15 minutes. Each sample was then weighed again, including the weight of the pan. The percent solid for each sample was then calculated. See Table X for percent solids results. One soil sample, C00159446, was weighed at ~8g for percent solid analysis, rather than at ~20g which is stated in the method. See deviation 6. The percent solid calculation was altered to reflect this change in mass. 6.3 Extraction Procedure for Water A 40 mL aliquot of the water sample was used for the extraction procedure. After fortification of appropriate samples, the samples were loaded onto a Cis SPE cartridge conditioned with 10 mL of methanol and 5 mL of water. The eluate was discarded. Approximately five milliliters of methanol was added to the cartridge. Five milliliters of Exygen Research Page 15 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 eluate was collected into a graduated 15 mL polypropylene centrifuge tube. Each sample was analyzed by LC/MS/MS electrospray. 6.4 Preparation of Standards and Fortification Solutions A mixed stock standard solution of PFOA and 13C PFOA was prepared at a concentration of 1000 pg/mL by dissolving 100 mg of each of the standards (corrected for purity and salt content) in 100 mL of methanol. From this solution, the following fortification standards were prepared: Cone, of Fort Stock or Fort. Volume Solution (mL) (pg/mL)1 1000 10 100 10 10 10 1.0 10 0.1 10 1of PFOA and 13C PFOA Final Volume (mL) 100 100 100 100 100 Final Cone, of Fortification Std. (pg/mL) 100 10 1.0 0.1 0.01 1 -5 A stock solution of only C PFOA was also prepared at a concentration of 100 pg/mL by dissolving 10 mg the standard (corrected for purity and salt content) in 100 mL of methanol. From this solution, the following fortification standards were prepared: Cone, of Stock or Fort. Solution (pg/mL)1 100 10 1.0 0.1 1of 13C PFOA Fort Volume (mL) 10 10 10 10 Final Volume (mL) 100 100 100 100 Final Cone, of Fortification Std. (pg/mL) 10 1.0 0.1 0.01 Exygen Research Page 16 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 A set of extracted calibration standards containing PFOA and 13C PFOA were also made. The following concentrations were prepared before extraction: Cone, of Fort Fort Solution Volume (ng/mL)1 (mL) - 10 0.1 10 0.2 10 0.4 100 0.1 100 0.2 100 0.4 1of PFOA and 13C PFOA Volume of Fortified Sample (mL) 40 40 40 40 40 40 40 Final Cone, of Calibration Std. (ng/mL) 0 25 50 100 250 500 1000 The stock standard solution and all fortification and calibration standard solutions were stored in a refrigerator (4 2C) when not in use. Documentation of standard preparation is located in the raw data package associated with this interim report. 6.5 Chromatography Quantification of PFOA was accomplished by LC/MS/MS electrospray. The retention time of PFOA was ~11.2 min. Peaks above the LOQ were not detected in any of the reagent blank samples corresponding to the analyte retention time. 6.6 Instrument Sensitivity The smallest standard amount injected during the chromatographic run had a concentration of 0.2 ng/g of PFOA for sediment samples and a concentration of 25 ng/L for water samples. 6.7 Description of LC/MS/MS Instrument and Operating Conditions Instrument: Interface: Computer: Software: HPLC: API 4000 Biomolecular Mass Analyzer Turbo Ion Spray Liquid Introduction Interface DELL OptiPlex GX400 Windows NT, Analyst 1.4.1 Hewlett Packard (HP) Series 1100 HP Quat Pump HP Vacuum Degasser HP Autosampler HP Column Oven Exygen Research Page 17 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 HPLC Column: Thermo Fluophase RP, 50 mm x 2.1 mm Column Temp.: ~30 C Injection Voi.: 15 pL Mobile Phase (A): 2 mM Ammonium Acetate in water Mobile Phase (B): Methanol Time /mini 0.0 1.0 8.0 10.0 11.0 18.0 %A 65 65 25 25 65 65 %B 35 35 75 75 35 35 Total run time: -18 min Flow Rate: 0.3 mL/min Ions monitored: Analvte PFOA PFOA Confirm Ion 13C PFOA Mode negative negative negative Transition Monitored 413 369 413->219 415 370 Approximate Retention Time (min) --11.2 min. -11.2 min. -11.2 min. 6.8 Quantitation and Example Calculation PFOA concentrations are quantitated in nanogram of PFOA per liter of extract and then converted to nanogram per gram. 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 of standards. The concentration was determined from the equations below. Equation 1 calculated the amount of analyte found (in ng/L, based on peak area) using the standard curve (linear regression parameters) generated by the Analyst software program. Equation 1: Analyte found (ng/L) = (Peak area - intercept) x DF Slope Where: DF = Dilution Factor, factor by which the final volume was diluted, if necessary. Exygen Research Page 18 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 For samples fortified with known amounts of PFOA and 13C PFOA prior to extraction, Equation 2 was used to calculate the percent recovery. Equation 2: Recovery (%) = (analyte found (ng/L) - analyte in control (ng/LV) xl00% amount added (ng/L) Note: For the PFOA recovery calculation, the "control" is the unspiked aliquot of the primary field sample. Equation 3 was used to convert the amount of PFOA found in ng/L to ng/g (ppb) in the soil samples. Equation 3: PFOA found (ppb) = fPFOA found (ng/L) x volume extracted (0.04D1 sample weight (5 g) Equation 4 was then used to calculate the amount of PFOA found in ppb based on dry weight. Equation 4: PFOA found (ppb) dry weight = PFOA found (ppb) x [100% / total solids(%)] NOTE: Total solids (%) = [wet weight (g) / dry weight (g)] x 100% An example of a calculation using an actual sample follows: Soil sample Exygen ID C0160874 Spk C (Set: 041706B), fortified at 500 ng/L with PFOA where: peak area = 121349 intercept = 18700 slope = 1060 dilution factor = 10 ng/L PFOA added (fort level) = 500 ng/L in corresponding sample (ng/L) = 502 volume extracted (L) = 0.04 sample weight (g) =5 total solids (%) = 87.24 Exygen Research Page 19 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 From equation 1: Analyte found (ng/L) = fl21349 - 187001 x 10 1060 = 968 ng/L From equation 2: % Recovery = (968 ng/L - 502 ng/Ll x 100% 500 ng/L = 93% From equation 3: PFOA found (ppb) = (968 ng/L x 0.04L) 5g = 7.74 ppb From equation 4: PFOA found (ppb) dry weight = 7.74 ppb x (100% / 87.24%) = 8.87 ppb Note: Numbers may vary slightly due to rounding. 7.0 EXPERIMENTAL DESIGN 13C PFOA was used as a surrogate for all of the samples. 13C PFOA was added to the soil samples in the laboratory after the samples were weighed for extraction. For soil samples designated as laboratory matrix spikes, PFOA was also added at a known concentration to the samples in the laboratory after the samples were weighed for extraction. For all o f the water samples, 13C PFOA was added to the sample collection bottles in the laboratory before being shipped to the field for sampling. For water samples designated as field matrix spikes, PFOA was also added at a known concentration to the bottles in the laboratory before being shipped to the field. The samples were filled to a 200 mL volumetric fill line in the field. The soil samples were extracted in thirty-two sets. Each set included one reagent blank and two reagent blanks fortified at known concentrations. Twenty-two of the sets contained five samples sites and one set contained two sample sites. One sample was collected for each sample site. For each site, two laboratory matrix spikes and a laboratory duplicate were extracted. The last eight sets were re-extraction sets. Four sets contained eight samples, three sets contained seven samples, and one set contained six samples. Each re-extraction set consisted of one sample and variable laboratory matrix spikes. Exygen Research Page 20 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 The water samples were extracted in two sets. Each set included one reagent blank and two reagent blanks fortified at known concentrations. Set one contained three rinse blanks, two trip blanks with the associated spikes. Set two contained four rinse blanks and two trip blanks with the associated spikes. 8.0 RESULTS Analytical results and assessed accuracies for the analysis of PFOA in soil samples based on dry weight are summarized in Table I. Analytical results and assessed accuracies for the analysis of PFOA in re-extracted soil samples based on dry weight are summarized in Table II. Analytical results and assessed accuracies for the analysis of PFOA in water samples are summarized in Table III. Field matrix spikes that were in the appropriate concentration range (primary concentration must be less than or equal to three times the spiking level) were used to calculate the assessed accuracies. When the field matrix spikes were not available or failed to meet recovery criteria, the laboratory spikes were used. Quantitative PFOA results were obtained for all samples except for nine soil samples that are not reported due to quality control failures. Fortification recoveries for the analysis of PFOA in soil samples based on wet weight are summarized in Table IV. The overall average percent recovery standard deviation for PFOA in soil samples based on wet weight was 80 19%. Fortification recoveries for the analysis of PFOA in re-extracted soil samples based on wet weight are summarized in Table V. The overall average percent recovery standard deviation for PFOA in re extracted soil samples based on wet weight was 74 18%. Fortification recoveries for the analysis of PFOA in water samples are summarized in Table VI. The overall average percent recovery standard deviation for PFOA in water samples was 92 7%. Fortification recoveries for 13C PFOA in soil samples based on wet weight are summarized in Table VII. The average percent recovery standard deviation for C PFOA in soil samples based on wet weight was 91 32%. Fortification recoveries for C PFOA in re-extracted soil samples based on wet weight are summarized in Table VIII. The average percent recovery standard deviation for 13C PFOA in re-extracted soil samples based on wet weight was 48 30%. Fortification recoveries for C PFOA in water samples are summarized in Table IX. The average percent recovery standard deviation for 13C PFOA in water samples was 107 14%. The total percent solids for soil samples are detailed in Table X. 9.0 CONCLUSIONS Except as noted above, the soil and water samples were successfully extracted and analyzed for PFOA according to analytical methods V0001781 and V0001780, Exygen Research Page 21 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 respectively. There were no circumstances that would have affected the quality or integrity of the data. 10.0 RETENTION OF DATAAND SAMPLES All original paper data generated by Exygen Research that pertains to this interim report will be shipped to the study director. This does not include facility-specific raw data such as instrument or temperature logs. Exact copies of all raw data, as well as a signed copy of the interim analytical report and all original facility-specific raw data, will be retained in the Exygen Research archives for the period of time specified in EPA TSCA Good Laboratory Practice Standards 40 CFR 792. Exygen Research Page 22 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 TABLES Exygen Research Page 23 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table I. Summary of PFOA in Soil Samples Analyte Found (ppb, ng/g) Dry Weight Client C8 Acid PFOA Sample ID_______________ Perfluorooctanoic Acid Assessed Accuracy (+/- %) C0156344 C0156344 Rep C0156345 C0156345 Rep C0156346 C0156346 Rep C0156347 C0156347 Rep C 0156351 C0156351 Rep C0156352 C0156352 Rep C0156353 C0156353 Rep C0156354 C0156354 Rep C0156355 C0156355 Rep C0156356 C0156356 Rep C0156357 C0156357 Rep C0156358 C0156358 Rep C0156360 C0156360 Rep C 0 156361 C0156361 Rep C0156362 C0156362 Rep D F13-SS-SS03-0-0000 D F13-S S -S S 03-0-0000* D F13-S S -S S 03-0-0003 D F13-S S -S S 03-0-0003* D F13-S S -S S 04-0-0000 D F13-S S -S S 04-0-0000* D F13-S S -S S 04-0-0003 D F13-S S -S S 04-0-0003* D F 13-S S -S S 05-0-0000 D F13-S S -S S 05-0-0000* D F13-S S -S S 05-0-0003 D F13-S S -S S 05-0-0003* DF 13-SS-SS 05-D B-0003 D F13-SS-SS05-D B-0003* D F10-SS-SS01-0-0000 D F10-SS-SS01-0-0000* D F10-SS-SS01-0-0003 D F10-SS-SS01-0-0003* D F10-S S -S S 02-0-0000 D F10-S S -S S 02-0-0000* DF 10-S S -S S 02-D B -0000 D F10-SS-SS02-D B-0000* D F10-S S -S S 02-0-0003 D F10-S S -S S 02-0-0003* D F10-S S -S S 03-0-0000 D F10-S S -S S 03-0-0000* D F10-S S -S S 03-0-0003 D F10-S S -S S 03-0-0003* D F10-S S -S S 04-0-0000 D F10-S S -S S 04-0-0000* NR NR NR NR 5 6 .7 5 8 .6 339 288 NR NR NR NR NR NR NR NR 191 220 119 116 105 108 9 9 .8 9 8 .9 1 6 .9 15.2 2 0 .3 2 0 .8 339 363 - - - - 40 40 40 40 - - - - - - . - 50 50 50 50 40 40 40 40 50 50 30 30 30 30 ' Laboratory Duplicate ND = Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR = Not Reported due to Quality Control failures, see Table II for re-extract data. Exygen Research Page 24 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table I. Summary of PFOA in Soil Samples Continued Analyte Found (ppb, ng/g) Dry Weight C8 Acid PFOA Perfluorooctanoic Acid Assessed Accuracy (+/- */) C0156363 C0156363 Rep C0156364 C0156364 Rep C0156365 C0156365 Rep C0156366 C0156366 Rep C0156367 C0156367 Rep C0156368 C0156368 Rep C0156370 C0156370 Rep C 0156371 C0156371 Rep C0156372 C0156372 Rep C0156373 C0156373 Rep C0156374 C0156374 Rep C0156375 C0156375 Rep C0156376 C0156376 Rep C0156377 C0156377 Rep C0156378 C0156378 Rep C0156379 C0156379 Rep C0156380 C0156380 Rep C 0 156381 C0156381 Rep D F10-SS-SS04-0-0003 D F10-S S -S S 04-0-0003* D F10-SS-SS05-0-0000 D F10-S S -S S 05-0-0000* D F10-S S -S S 05-0-0003 D F10-S S -S S 05-0-0003* D F 11-S S -S S 01-0-0000 D F11-SS-SS01-0-0000* D F11-SS-SS01-0-0003 D F11-SS-SS01-0-0003* D F11-SS-SS02-0-0000 D F 1 1 -S S -S S 0 2 -0 -0 0 0 0 * D F11-S S -S S 02-0-0003 D F11-SS-SS02-0-0003* D F11-SS-SS03-0-0000 D F11-S S -S S 03-0-0000* D F 11-S S -S S 03-D B -0000 D F 11-S S -S S 03-D B -0000* D F 11-S S -S S 03-0-0003 D F11-S S -S S 03-0-0003* D F 11-S S -S S 04-0-0000 D F11-S S -S S 04-0-0000* D F 11-S S -S S 04-0-0003 D F11-S S -S S 04-0-0003* D F 11-S S -S S 05-0-0000 D F11-S S -S S 05-0-0000* D F11-SS-SS05-0-0003 D F11-SS-SS05-0-0003* D F05-SS-SS01-0-0000 D F05-SS-SS01-0-0000* D F05-SS-SS01-0-0003 D F05-SS-SS01-0-0003* D F05-S S -S S 02-0-0000 D F05-S S -S S 02-0-0000* D F05-S S -S S 02-0-0003 D F05-S S -S S 02-0-0003* 387 435 338 313 699 560 257 250 NR NR NR NR NR NR NR NR 138 170 NR NR 2 8 .7 3 2 .2 102 9 7 .6 48 .9 4 7 .4 5 3 .4 5 8 .5 119 121 5 1 .6 40 .5 346 366 NR NR 30 30 50 50 50 50 30 30 - - - - 50 50 - 40 40 40 40 30 30 40 40 50 50 40 40 30 30 - - ` Laboratory Duplicate ND = Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR = Not Reported due to Quality Control failures, see Table II for re-extract data. Exygen Research Page 25 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table I. Summary of PFOA in Soil Samples Continued E xyg e n ID C0156382 C0156382 Rep CO156383 C0156383 Rep CO156384 C0156384 Rep CO156385 CO156385 Rep CO156386 C0156386 Rep CO156387 CO156387 Rep C0156388 C0156388 Rep CO156389 C0156389 Rep CO156390 C0156390 Rep C0156391 C0156391 Rep CO156392 C0156392 Rep C0156393 CO156393 Rep C0156394 CO156394 Rep C0156395 CO156395 Rep C0159446 C0159446 Rep C0159447 C0159447 Rep C lie n t S a m p le ID DF 12-S S -S S 01-0-0000 D F12-SS-SS01-0-0000* DF 12-S S -S S 01-0-0003 DF 12-S S -S S 01-0-0003* D F12-SS-SS02-0-0000 D F12-SS-SS02-0-0000* D F12-SS-SS02-0-0003 D F12-SS-SS02-0-0003* D F12-SS-SS03-0-0000 DF 12-S S -S S 03-0-0000* DF 12-S S -S S 03-0-0003 DF 12-S S -S S 03-0-0003* D F12-SS-SS04-0-0000 D F12-S S -S S 04-0-0000* DF 12-S S -S S 04-0-0003 DF 12-S S -S S 04-0-0003* DF 12-S S -S S 05-0-0000 DF 12-S S -S S 05-0-0000* DF 12-S S -S S 05-0-0003 DF 12-S S -S S 05-0-0003* DF 13-S S -S S 01-0-0000 DF 13-S S -S S 01-0-0000* DF 13-S S -S S 01-0-0003 D F13-S S -S S 01-0-0003* D F13-SS-SS02-0-0000 D F13-SS-SS02-0-0000* D F13-SS-SS02-0-0003 DF 13-S S -S S 02-0-0003* D A L-SB-603R -0-0000 D A L-SB-603R -0-0000* D A L-SB-603R -0-0005 D A L-SB-603R -0-0005* A n a ly te F o u n d (p p b , n g /g ) D ry W e ig h t C 8 A c id PFO A P e rflu o ro o c ta n o ic A c id 4 9 .8 8 2 .8 NR NR NR NR NR NR NR NR 2 9 .9 4 0 .7 NR NR NR NR 42.1 4 8 .8 176 174 310 315 NR NR NR NR NR NR NR NR 4 .0 2 4 .0 4 Assessed A ccuracy ( /-% ) 30 30 - - - - - 50 50 - - 30 30 40 40 50 50 - - - - - " - 50 50 "Laboratory Duplicate ND = Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR = Not Reported due to Quality Control failures, see Table II for re-extract data. Exygen Research Page 26 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table I. Summary of PFOA in Soil Samples Continued Client Exygen ID__________ Sample ID C0159448 C0159448 Rep C0159449 C0159449 Rep C0159450 C0159450 Rep C 0159451 C0159451 Rep C0159453 CO159453 Rep C0159454 C0159454 Rep C0159455 C0159455 Rep C0159456 C0159456 Rep C0159457 C0159457 Rep C0159458 C0159458 Rep C0159459 C 0159459 Rep C0159460 C 0159460 Rep C 0 159461 C0159461 Rep CO159462 C0159462 Rep C0159463 C0159463 Rep C0159464 C0159464 Rep C0159466 C0159466 Rep C0159467 C0159467 Rep C0159468 C0159468 Rep D A L-S B -603R -0-0010 D A L-S B -603R -0-0010* D AL-SB-603R -0-0050 D A L-SB-603R -0-0050* D AL-SB-603R -0-0100 D A L-S B -603R -0-0100* D A L-SB-603R -D B-0100 D AL-SB-603R -D B-0100* D AL-SB-604R -0-0000 D A L-SB-604R -0-0000* D A L-SB-604R -0-0005 D A L -S B -6 04R -0 -00 05* D A L-S B -604R -0-0010 D A L-S B -604R -0-0010* D A L-SB-604R -0-0050 D A L-S B -604R -0-0050* D A L-S B -604R -0-0100 D A L-S B -604R -0-0100* D A L-SB-605R -0-0000 D A L-S B -605R -0-0000* D AL-SB-605R -D B-0000 D AL-SB-605R -D B-0000* D A L-SB-605R -0-0005 D A L-SB-605R -0-0005* D A L-S B -605R -0-0010 D A L-S B -605R -0-0010* D A L-SB-605R -0-0050 D A L-SB-605R -0-0050* D A L-S B -605R -0-0100 D A L-S B -605R -0-0100* D AL-SB-138R -0-0000 D A L-SB-138R -0-0000* D A L-SB-138R -0-0005 D A L-SB-138R -0-0005* D A L-SB-138R -0-0010 D A L-S B -138R -0-0010* D A L-SB-138R -0-0050 D A L-S B -138R -0-0050* Analyte Found (ppb, ng/g) Dry Weight C8 Acid PFOA Perfluorooctanoic Acid 5 .3 4 4 .7 8 NR NR 1.45 1.10 0 .9 3 6 0 .9 9 0 NR NR NR NR 1.46 1.62 5.71 5 .5 2 ND 0 .3 5 6 7 .9 5 7 .8 6 7.61 7 .9 8 7 .8 2 7 .4 6 2 .6 4 3 .3 3 1.01 1.03 0 .8 4 9 0 .8 6 2 2 7 .6 29.1 4 8 .8 5 7 .2 4 0 .7 4 7 .4 NR NR Assessed Accuracy (+/- */) 40 40 - 40 40 30 30 - . 30 30 30 30 30 30 40 40 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 _ ' Laboratory Duplicate ND = Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR = Not Reported due to Quality Control failures, see Table II for re-extract data. Exygen Research Page 27 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table I. Summary of PFOA in Soil Samples Continued Analyte Found (ppb, ng/g) Dry Weight Client C8 Acid PFOA Exyqen ID___________ Sample ID_______________ Rerfluorooctanolc Acid Assessed Accuracy C0159470 C 0159470 Rep C 0 159471 C0159471 Rep C0160871 C0160871 Rep CO160872 CO160872 Rep CO160873 C0160873 Rep C0160874 CO160874 Rep C0160875 C0160875 Rep C 0160876 CO160876 Rep CO160877 CO160877 Rep C0160878 C0160878 Rep C0160879 C0160879 Rep CO160880 C0160880 Rep C0160881 C0160881 Rep CO160882 CO160882 Rep D A L-S B -138R -0-0100 D A L-S B -138 R -0 -0100* D AL-SB-138R -D B -0100 D A L-S B -138R -D B -0100* D F05-S S -S S 03-0-0000 D F05-SS-SS03-0-0000* D F05-SS-SS03-0-0003 D F05-S S -S S 03-0-0003* D F05-SS-SS04-0-0000 D F05-SS-SS04-0-0000* D F05-SS-SS04-0-0003 D F 0 5 -S S -S S 0 4 -0 -0 0 0 3 * D F05-SS-SS05-0-0000 D F05-SS-SS05-0-0000* D F05-SS-SS05-0-0003 D F05-S S -S S 05-0-0003* D F05-SS-SS05-D B-0003 D F05-SS-SS05-D B-0003* D F09-S S -0006-0-0000 D F09-S S -0006-0-0000* D F09-SS-0006-0-0003 D F09-S S -0006-0-0003* D F09-S S -0006-0-0010 D F09-S S -0006-0-0010* D F09-S S -0007-0-0000 D F09-S S -0007-0-0000* D F09-SS-0007-0-0003 D F09-S S -0007-0-0003* 354 269 308 309 NR NR NR NR NR NR 4 .6 0 4 .6 2 5 .7 9 5.71 7 .2 3 6 .7 6 7.41 7 .1 2 14.8 14.9 16.3 15.4 8.91 8 .9 2 173 182 124 112 40 40 30 30 - - . - - 30 30 40 40 30 30 50 50 30 30 30 30 40 40 50 50 50 50 ' Laboratory Duplicate ND = Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR = Not Reported due to Quality Control failures, see Table II for re-extract data. Exygen Research Page 28 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table I. Summary of PFOA in Soil Samples Continued Exygen ID C0160883 CO160883 Rep C0160884 C0160884 Rep CO160885 CO160885 Rep C0160886 CO160886 Rep CO160887 CO160887 Rep CO160888 CO160888 Rep CO160889 CO160889 Rep CO160890 CO160890 Rep C0160891 C0160891 Rep CO160892 C0160892 Rep C0160893 CO160893 Rep CO160894 CO160894 Rep CO160895 C 0160895 Rep CO160896 CO160896 Rep Client Sample ID D F09-S S -0007-0-0010 D F09-S S -0007-0-0010* D F09-S S -0007-D B -0010 D F09-S S -0007-D B -0010* D F09-S S -0008-0-0000 D F09-S S -0008-0-0000* D F09-S S -0008-0-0003 D F09-S S -0008-0-0003* D F09-S S -0008-0-0010 D F09-S S -0008-0-0010* D F09-SS-0009-0-0000 D F 0 9 -S S -0 0 0 9 -0 -0 0 0 0 * D F09-S S -0009-0-0003 D F09-S S -0009-0-0003* D F09-SS-0009-D B-0003 D F09-SS-0009-D B-0003* D F09-S S -0009-0-0010 D F09-S S -0009-0-0010* D F09-SS-0010-0-0000 D F09-SS-0010-0-0000* D F09-SS-0010-0-0003 D F09-SS-0010-0-0003* D F09-S S -0010-0-0010 D F09-S S -0010-0-0010* D F09-S S -0011-0-0000 D F 09-S S -0011-0-0000* D F 09-S S -0011-0-0003 D F 09-S S -0011-0-0003* Analyte Found (ppb, ng/g) Dry Weight C8 Acid PFOA Perfluorooctanoic Acid 4 6 .7 5 1 .4 3 8 .9 4 7 .9 176 149 149 135 122 108 10.1 11.1 12.2 14.6 12.3 13.5 19.0 23.1 886 934 511 498 461 357 253 297 284 360 Assessed Accuracy <+/-% ) 50 50 30 30 30 30 40 40 30 30 40 40 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 'Laboratory Duplicate ND = Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR = Not Reported due to Quality Control failures, see Table II for re-extract data. Exygen Research Page 29 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table I. Summary of PFOA in Soil Samples Continued Exygen ID Client Sample ID Analyte Found (ppb, ng/g) Dry Weight C8 Acid PFOA Perfluorooctanoic Acid Assessed Accuracy (+/-/.) C0160897 C0160897 Rep C0160898 C0160898 Rep C0160899 C0160899 Rep C0160900 C0160900 Rep C 0160901 C0160901 Rep C0160902 C0160902 Rep C0160903 C0160903 Rep CO160904 C0160904 Rep CO164247 C0164247 Rep C0164248 C0164248 Rep C 0164249 C0164249 Rep CO164250 CO164250 Rep C 0 164251 C0164251 Rep C0164252 C0164252 Rep C0164253 C0164253 Rep C0164254 C0164254 Rep D F09-S S -0011-0-0010 D F09-S S -0011-0-0010* D F09-SS-0012-0-0000 D F09-SS-0012-0-0000* D F09-SS-0012-D B-0000 D F 09-S S -0012-D B -0000* D F09-SS-0012-0-0003 D F09-SS-0012-0-0003* D F09-SS-0012-0-0010 D F09-SS-0012-0-0010* D F09-SS-0013-0-0000 D F09-S S -0013-0-0000* D F09-S S -0013-0-0003 D F09-SS-0013-0-0003* D F09-S S -0013-0-0010 D F09-S S -0013-0-0010* D A L-SB-602R -0-0000 D A L-S B -602R -0-0000* D A L-SB-602R -0-0005 D A L-SB-602R -0-0005* D A L-S B -602R -0-0010 D A L-S B -602R -0-0010* D A L-SB-602R -0-0050 D A L-S B -602R -0-0050* D A L-S B -602R -0-0100 D A L-S B -602R -0-0100* D AL-SB-601R -0-0000 D A L-SB-601R -0-0000* D AL-SB-601 R -D B-0000 D AL-SB-601 R -D B-0000 DAL-SB-601 R -0-0005 DAL-SB-601 R -0-0005 NR NR NR NR 535 500 NR NR 481 454 21.1 2 1 .0 2 5 .4 2 9 .3 5 0 .5 4 2 .7 6.11 6 .5 8 6.91 6 .7 3 4 .0 3 3 .2 4 0 .7 6 6 0 .6 7 4 0 .6 6 5 0.561 1 .7 5 2.11 2 .2 2 1 .9 7 1.91 1 .8 7 - - - - 30 30 - - 40 40 30 30 30 30 40 40 30 30 30 30 30 30 30 30 40 40 30 30 30 30 30 30 'Laboratory Duplicate ND = Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR = Not Reported due to Quality Control failures, see Table II for re-extract data. Exygen Research Page 30 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table I. Summary of PFOA in Soil Samples Continued Bt^genJD C0164255 C0164255 Rep C0164256 C0164256 Rep C0164257 C0164257 Rep C0164259 C0164259 Rep C0164260 C0164260 Rep Client Analyte Found (ppb, nglg) Dry Weight C8 Acid PFOA Assessed Accuracy D AL-SB-601R -0-0010 D AL-SB-601R -0-0010 D AL-SB-601R -D B-0010 D AL-SB-601R -D B-0010 D AL-SB-601 R -0-0050 D AL-SB-601 R -0-0050 D AL-SB-601 R -0-0100 D AL-SB-601 R -0-0100* DAL-SB-601 R -D B -0100 D AL-SB-601 R -D B -0100* NR NR 2 .1 5 1 .6 6 0 .8 9 5 1 .4 2 0 .4 7 7 0 .4 3 8 0 .4 8 5 0 .3 9 9 - 40 40 30 30 30 30 30 30 'Laboratory Duplicate ND = Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR = Not Reported due to Quality Control failures, see Table II for re-extract data. Exygen Research Page 31 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table II. Summary of PFOA in Re-extracted Soil Samples Exygen ID C0156344 C0156345 C 0 156351 C0156352 C0156353 C0156354 C0156367 C0156368 C0156370 C 0156371 C0156373 C 0 1 56381 C0156383 C0156384 C 0156365 C0156386 C0156388 C0156389 C0156393 C0156394 C0156395 C0159449 C0159453 C0159454 C0159468 C0160871 C0160872 C0160873 C 0160897 C 0160898 C0160900 C 0164255 Client Sample ID D F 1 3 -S S -S S 0 3 -0 -0 0 0 0 D F 1 3 -S S -S S 0 3 -0 -0 0 0 3 D F13-SS-SS05-0-0000 D F13-SS-SS05-0-0003 D F 1 3 -S S -S S 0 5 -D B -0 0 0 3 D F 1 0 -S S -S S 0 1 -0 -0 0 0 0 D F 1 1 -S S -S S 0 1 -0 -0 0 0 3 D F 1 1-S S -S S 0 2 -0 -0 0 0 0 DF 11-SS-SS02-0-0003 D F 1 1 -S S -S S 0 3 -0 -0 0 0 0 DF 11-SS-SS03-0-0003 D F 0 5 -S S -S S 0 2 -0 -0 0 0 3 D F 1 2 -S S -S S 0 1 -0 -0 0 0 3 D F 1 2 -S S -S S 0 2 -0 -0 0 0 0 D F 1 2 -S S -S S 0 2 -0 -0 0 0 3 DF 12-SS-SS03-0-0000 D F 1 2 -S S -S S 0 4 -0 -0 0 0 0 D F 1 2 -S S -S S 0 4 -0 -0 0 0 3 D F 1 3 -S S -S S 0 1 -0 -0 0 0 3 D F 1 3 -S S -S S 0 2 -0 -0 0 0 0 D F 1 3 -S S -S S 0 2 -0 -0 0 0 3 D AL-SB-603R-0-0050 D AL-SB-604R-0-0000 D AL-SB-604R-0-0005 D AL-SB-138R-0-0050 D F 0 5 -S S -S S 0 3 -0 -0 0 0 0 D F 0 5 -S S -S S 0 3 -0 -0 0 0 3 D F 0 5 -S S -S S 0 4 -0 -0 0 0 0 D F09-SS-0011-0-0010 D F 0 9 -S S -0 0 1 2 -0 -0 0 0 0 D F 0 9 -S S -0 0 1 2 -0 -0 0 0 3 D A L -S B -6 0 1 R -0 -0 0 1 0 Analyte Found (ppb, ng/g) Dry Weight C8 Acid PFOA Perfluorooctanoic Acid 525 551 1130 1020 857 NR 8 6 .8 NR 818 102 NR 335 197 2140 3910 2 8 .6 4390 11900 561 3350 5930 3.71 2 .2 9 3 .7 5 104 227 NR NR 238 NR NR NR Assessed Accuracy <+/-%> 40 30 40 30 50 30 30 40 50 30 50 40 50 30 30 30 50 30 30 50 30 30 40 40 - - NO = Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g. NR = Not reported due to quality control failures. Exygen Research Page 32 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table III. Summary of PFOA in Associated Rinse Blanks and Trip Blanks Exygen ID C0156348 C0156359 C0156369 C0159443 C0159452 C0159465 C0159469 C0160867 C0160870 CO164222 CO164258 C lient Sam ple ID DF13-SS-TRIP1 -0-060221 D F 1 0-S S -S S 0 3-R B -0 000 D F 1 1-S S -S S 0 2-R B -0 003 DAL-SB-TRIP1-0-060302 D A L -S B -6 03R -R B -0100 DAL-SB-138R-RB-0000 DAL-SB-138R-RB-0100 D F 05-S S -T R IP 2-0-060308 D F05-S S -S S 03-R B -0000 D A L -S B -T R IP -0-0 603 16 DAL-SB-601R-RB-0100 Analyte Found (ppt, ng/L) C 8A cid PFOA Ptrfluorooctanolc Add ND ND ND ND ND ND ND ND ND ND ND Assessed Accuracy <+/-%> 30 30 30 30 30 30 30 30 30 30 30 ND - Not detected at or above the Limit of Quantitation (LOQ) of 25 ng/L. Exygen Research Page 33 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table IV. Matrix Spike Recovery of PFOA in Soil Samples Sam ple Description D F 1 3-S S -S S 0 3-0-000 0 (C0155344 Spk C. 4 ppb Spile) D F 1 3-S S -S S 0 3-0-000 0 (C0156344 Spk D, 400 ppb Splk) D F 1 3-S S -S S 0 3-0-000 3 (C0156345 Spk E, 4 ppb Splk) D F 1 3-S S -S S 0 3-0-000 3 (C015C345 Spk F, 400 ppb Splk) D F 1 3-S S -S S 0 4-0-000 0 (C0156346 Spk G, 4 ppb Splk) D F 1 3-S S -S S 0 4-0-000 0 (C0155346 Spk H, 400 ppb Spika) D F 1 3-S S -S S 0 4-0-000 3 (C0156347 Spk 1, 4 ppb Splk) D F 1 3-S S -S S 0 4 -0 -0 0 0 3 (C0156347 Spk J, 400 ppb Splk) D F 1 3-S S -S S 0 5-0-000 0 (C0155351 Spk K, 4 ppb Splk) D F 1 3-S S -S S 0 5-0-000 0 (00150351 Spk L, 400 ppb Splk) D F 1 3 -S S -S S 0 5 -0 -0 0 0 3 (C0150352 Spk C. 4 ppb Splk) D F 1 3-S S -S S 0 5-0-000 3 (C0150352 Spk D, 400 ppb Splk) D F 1 3-S S -S S 0 5-D B -0 003 (C0150353 Spk E, 4 ppb Splk) D F 1 3-S S -S S 0 5-D B -0 003 (C0150353 Spk F, 400 ppb Splk) D F 10-SS-SS01-0-0000 (00150354 Spk G, 4 ppb Splk) D F10-S S -S S 01-0-00 0 0 (C0156354 Spk H, 400 ppb Spike) DF10-SS-SS01 -0-0003 (00150355 Spk 1,4 ppb Splk) D F 1 0-S S -S S 0 1-0-000 3 (C0150355 Spk J, 400 ppb Splk) D F 1 0-S S -S S 0 2-0-000 0 (C0150350 Spk K. 4 ppb Splk) D F 1 0-S S -S S 0 2-0-000 0 (C0150350 Spk L, 400 ppb Splk) Amount Spiked (ng/g) 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 Amt Found In Sample (ng/g) wet weight C8 Acid PFOA Amount Recovered (ng/g) wet weight NR NR NR NR NR NR NR NR 47.3 47.3 52.7 298 296 284 296 570 NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR 166 161 166 397 102 105 102 330 Recovery (% )' NR NR NR NR * 63 69 NR NR NR NR NR NR NR NR * 58 57 `Sample residue exceeds the spiking level significantly (3x spiking level); therefore, an accurate recovery value cannot be calculated. ND = Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR = Not Reported due to Quality Control bilures, see Table V for re-extract data. Note: Since this summary table shows rounded results, recovery values may vary slightly from the values In the raw data. Exygen Research Page 34 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table IV. Matrix Spike Recovery of PFOA in Soil Samples Continued Sample Description D F 1 0-S S -S S 0 2-D B -0 000 (C0156307 Spk C, 4 ppb Spike) D F10-S S -S S 0 2-D B -0 000 (C0106307 Spk D, 400 ppb Spike) D F 1 0-S S -S S 0 2-0-000 3 (C010S3SO Spk E, 4 ppb Spike) D F 1 0-S S -S S 0 2-0-000 3 (C0100306 Spk F, 400 ppb Spike) D F10-SS-SS03-0-0000 (C01M3M Spk G, 4 PPb Spike) D F 1 0-S S -S S 0 3-0-000 0 (C0106360 Spk H. 400 ppb Spike) D F 1 0-S S -S S 0 3-0-000 3 (C0156341 Spk 1,4 ppb Spike) D F 1 0-S S -S S 0 3-0-000 3 (C015S361 Spk J, 400 ppb Spike) D F 1 0-S S -S S 0 4-0-000 0 (C0100302 Spk K, 4 ppb Spike) D F 1 0-S S -S S 0 4-0-000 0 (C0150302 Spk L, 400 ppb Spike) D F 1 0-S S -S S 0 4 -0 -0 0 0 3 (C0150303 Spk C, 4 ppb Spike) D F 1 0-S S -S S 0 4-0-000 3 (CC01MM3 Spk 0 ,400 ppb Spike) D F 1 0-S S -S S 0 5-0-000 0 (C0150304 Spk E, 4 ppb Spike) D F 1 0-S S -S S 0 5-0-000 0 (C0150304 Spk F, 400 ppb Spike) D F 1 0-S S -S S 0 5-0-000 3 (C01M36S Spk G, 4 ppb Spike) D F 1 0-S S -S S 0 5-0-000 3 (C015830S Spk H. 400 ppb Spike) D F 1 1-S S -S S 0 1-0 -0 0 0 0 (C0100300 Spk 1.4 ppb Spike) DF11-SS-SS01 -0-0000 (C01503M Spk J, 400 ppb Spike) DF 11 -SS-SS01-0-0003 (C01MM7 Spk K, 4 ppb Spike) D F 11-S S -S S 01-0-0003 (C0150M7 Spk U 400 ppb Spike) Amount Spiked (ng/g) 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 Amt Found in Sample (ng/g) wet weight C8 Acid PFOA Amount Recovered (ng/g) wet weight 91.2 01.2 86.4 86.4 13.8 13.8 17.1 17.1 242 242 293 293 102 364 62.4 330 14.8 246 19.4 304 196 530 310 598 278 263 278 490 577 596 577 800 209 228 209 498 NR NR NR NR Recovery (% ) * 68 * 61 58 * 72 72 76 53 56 72 NR NR `Sample residue exceeds the spiking level significantly (3x spiking level); therefore, an accurate recovery value cannot be calculated. ND = Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR = Not Reported due to Quality Control failures, see Table V for re-extract data. 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 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table IV. Matrix Spike Recovery of PFOA in Soil Samples Continued Sam ple Description D F 1 1-S S -S S 0 2-0 -000 0 (C015S3SS Spk C, 4 ppb Spike) D F11-SS-SS02-0-0000 (C0156SM Spk D, 400 ppb Spike) D F11-SS-SS02-0-0003 (C0150370 Spk E, 4 ppb Spike) D F 1 1-S S -S S 0 2-0-000 3 (C0155370 Spk F, 400 ppb Spike) D F 1 1-S S -S S 0 3 -0 -0 0 0 0 (C0156371 Spk G, 4 ppb Spike) D F 1 1-S S -S S 0 3 -0 -0 0 0 0 (C0156371 Spk H, 400 ppb Spike) D F 1 1-S S -S S 0 3-D B -0 000 (C01SSS72 Spk 1,4 ppb Spike) D F11-SS-SS03-DB-0000 (C0150372 Spk J, 400 ppb Spike) D F 1 1-S S -S S 0 3 -0 -0 0 0 3 (C0150373 Spk K. 4 ppb Spike) D F 1 1-S S -S S 0 3 -0 -0 0 0 3 (C0150373 Spk Lt 400 ppb Spike) D F11-SS-SS04-0-0000 (C0150374 Spk C, 4 ppb Spike) D F 1 1-S S -S S 0 4 -0 -0 0 0 0 (C0150374 Spk D. 400 ppb Spike) D F 1 1-S S -S S 0 4 -0 -0 0 0 3 (C0150375 Spk E, 4 ppb Spike) D F11-SS-SS04-0-0003 (C0150375 Spk F, 400 ppb Spike) D F 1 1-S S -S S 0 5-0-000 0 (C0150370 Spk G, 4 ppb Spike) D F 11-SS-SS05-0-0000 (C0156376 Spk H, 400 ppb Spike) D F 1 1-S S -S S 0 5-0 -000 3 (C0150377 Spk 1,4 ppb Spike) D F 1 1-S S -S S 0 5-0-000 3 (C0150377 Spk J, 400 ppb Spike) DF05-SS-SS01-0-0000 (C0150378 Spk K, 4 ppb Spike) DF05-SS-SS01-0-0000 (C0150378 Spk L, 400 ppb Spike) Amount Spiked (ng/g) 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 Amt Found in Sample (ng/g) wet weight C8 Acid PFOA Amount Recovered (ng/g) wet weight NR NR NR NR NR NR NR NR NR NR NR NR 110 113 110 316 NR NR NR NR 24.9 24.9 91.2 91.2 40.9 40.9 29.4 298 96.0 342 34.6 322 45.5 45.5 44.9 317 96.8 96.8 126 318 Recovery (% ) NR NR NR NR NR NR * 52 NR NR 68 63 * 70 68 55 `Sample residue exceeds the spiking level significantly (3x spiking level); therefore, an accurate recovery value cannot be calculated. ND = Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR = Not Reported due to Quality Control bilures, see Table V for re-extract data. 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 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table IV. Matrix Spike Recovery of PFOA in Soil Samples Continued Sam ple Description D F 0 5-S S -S S 0 1-0 -0 0 0 3 (00156379 Spk C. 4 ppb S pilo) D F 0 5-S S -S S 0 1-0 -0 0 0 3 (C0156379 Spk D, 400 ppb S plkt) D F 0 5-S S -S S 0 2-0-000 0 (C0156380 Spk E, 4 ppb Splke) D F 0 5-S S -S S 0 2-0-000 0 (COI50380 Spk F, 400 ppb S plkt) D F 0 5-S S -S S 0 2-0-000 3 (C0156361 Spk G, 4 ppb Splka) D F 0 5-S S -S S 0 2-0-000 3 (COI59381 Spk H, 400 ppb Splka) D F 1 2-S S -S S 0 1-0 -0 0 0 0 (COI56382 Spk 1,4 ppb S plkt) D F 1 2-S S -S S 0 1-0 -0 0 0 0 (C0156362 Spk J, 400 ppb S plkt) D F 1 2-S S -S S 0 1-0-000 3 (COI56383 Spk K, 4 ppb Splke) D F 1 2-S S -S S 0 1-0 -0 0 0 3 (COI56383 Spk L, 400 ppb Splke) D F 12-S S -S S 02-0-0000 (C0156384 Spk C, 4 ppb Splke) D F 12-S S -S S 0 2 -0 -0 0 0 0 (COI56384 Spk D, 400 ppb S plkt) D F 1 2-S S -S S 0 2-0-000 3 (C015636S Spk E, 4 ppb S plkt) D F 1 2 -S S -S S 0 2 -0 -0 0 0 3 (COI56365 Spk F, 400 ppb Splke) D F 1 2-S S -S S 0 3-0-000 0 (COI56366 Spk G 4 ppb S p lkt) D F 1 2-S S -S S 0 3-0-000 0 (C0156366 Spk H, 400 ppb Splke) D F 1 2-S S -S S 0 3-0-000 3 (C0156387 Spk 1,4 ppb S plkt) D F 1 2-S S -S S 0 3-0-000 3 (C0156387 Spk J, 400 ppb S plkt) D F 1 2-S S -S S 0 4-0-000 0 (C0156388 Spk K, 4 ppb S plkt) D F 1 2-S S -S S 0 4-0-000 0 (C0156368 Spk L, 400 ppb S plkt) Amount Spiked (ng/g) 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 Amt Found in Sample (ngla) wet weight C8 Acid PFOA Amount Recovered (ng/g) wet weight 43.4 43.4 45.0 303 285 410 285 681 NR NR NR NR 41.2 41.2 80.8 326 NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR 26.2 26.2 22.3 254 NR NR NR NR Recovery (% ) * 65 99 NR NR * 71 NR NR NR NR NR NR NR NR 57 NR NR ` Sample residue exceeds the spiking level significantly (3x spiking level); therefore, an accurate recovery value cannot be calculated. ND = Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR - Not Reported due to Quality Control failures, see Table V for reextract data. 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 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table IV. Matrix Spike Recovery of PFOA in Soil Samples Continued Sam ple Description D F12-SS-SS04-0-0003 (C0156389 Spk C, 4 ppb S pila) D F 1 2-S S -S S 0 4-0-000 3 (C0156389 Spk D, 400 ppb Splk) D F12-SS-SS05-0-0000 (COI56390 Spk E, 4 ppb S pila) D F 1 2-S S -S S 0 5-0-000 0 (C0156390 Spk F, 400 ppb Splk) D F 1 2-S S -S S 0 5-0-000 3 (C0156391 Spk G 4 ppb S pila) D F 1 2-S S -S S 0 5-0-000 3 (C0156391 Spk H, 400 ppb S pila) D F 1 3-S S -S S 0 1-0-000 0 (C0156392 Spk 1, 4 ppb Splk) D F 1 3-S S -S S 01-0-0000 (C0156392 Spk J, 400 ppb Splk) D F 1 3-S S -S S 0 1-0-000 3 (C0156393 Spk K, 4 ppb S pila) D F 1 3-S S -S S 0 1-0-000 3 (C0156393 Spk L, 400 ppb Splke) D F 1 3-S S -S S 0 2-0-000 0 (C01SS3S4 Spk C, 4 ppb Splke) D F 1 3-S S -S S 0 2-0-000 0 (C0156394 Spk D, 400 ppb S pila) D F 1 3-S S -S S 0 2-0-000 3 (C0156395 Spk E, 4 ppb Splk) D F 1 3-S S -S S 0 2-0-000 3 (COI59395 Spk F, 400 ppb S pila) D A L-S B -603R -0-0000 (COI59446 Spk E, 4 ppb S pila) D A L -S B -6 03R -0 -00 00 (C0159446 Spk F, 400 ppb S pila) D A L -S B -6 03R -0 -00 05 (C015S447 Spk 0 ,4 ppb Splke) D A L -S B -6 03R -0 -00 05 (COI59447 Spk H, 400 ppb S pila) DAL-SB-603R-0-0010 (C0159446 Spk 1,4 ppb S pila) DAL-SB-603R-0-0010 (C0159449 Spk J. 400 ppb Splk) Amount Spiked (ng/g) 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 Amt Found in Sample (ng/g) wet weight C8 Acid PFOA Amount Recovered (ng/g) wet weight NR NR NR NR 33.0 33.0 37.3 406 143 148 143 407 240 251 240 438 NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR 3.45 3.45 4.36 4.36 5.57 268 6.81 282 Recovery (% ) NR NR 93 * 66 50 NR NR NR NR NR NR NR NR 53 66 61 69 ' Sample residue exceeds the spiking level significantly (3x spiking level); therefore, an accurate recovery value cannot be calculated. NO = Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR = Not Reported due to Quality Control failures, see Table V for re-extract data. Note: Since this summary table shows rounded results, recovery values may vary slightly from the values in the taw data. Exygen Research Page 38 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table IV. Matrix Spike Recovery of PFOA in Soil Samples Continued Sam ple Description D A L -S B -6 03R -0 -00 50 (C0159449 Spk K, 4 ppb Spike) D A L -S B -6 03R -0 -00 50 (C01SM4S Spk L, 400 ppb Spike) D A L-S B -603R -0-0100 (C01M4SOSpk C. 4 ppb Spike) D A L-S B -803R -0-0100 (C015S450Spk D, 400 ppb Spike) D A L -S B -6 03R -D B -0 100 (C01S04S1 Spk E, 4 ppb Spike) D A L -S B -603R -D B -0100 (C0159451 spk F, 400 ppb Spike) D A L-S B -604R -0-0000 (C01S04SS Spk G, 4 ppb Spike) D A L-S B -604R -0-0000 (C0199453 Spk H, 400 ppb Spike) D A L-S B -604R -0-0005 (C0159454 Spk 1,4 PPb Spike) D A L -S B -604R -0-0005 (C01S94S4Spk J. 400 ppb Spike) D A L -S B -6 04R -0 -0010 (C01S9455 Spk K, 4 ppb Spike) D A L -S B -604R -0-0010 (C0159455 Spk L, 400 ppb Spike) D A L-S B -604R -0-0050 (C0159456 Spk C, 4 ppb Spike) D A L-S B -604R -0-0050 (C0159456 Spk D, 400 ppb Spike) D A L -S B -6 04R -0 -0100 (C0199457 Spk E, 4 ppb Spike) D A L-S B -604R -0-0100 (C01594S7Spk F. 400 ppb Spike) D A L -S B -6 05R -0 -00 00 (C0159458 Spk G 4 ppb Spike) D A L -S B -6 05R -0 -00 00 (C015945S Spk H, 400 ppb Spike) DAL-SB-605R-DB-0000 (C01994S9 Spk 1,4 ppb Spike) DAL-SB-605R-DB-0000 (C0159459 Spk J, 400 ppb Spike) Amount Spiked (ng/g) 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 Amt Found in Sample (ng/g) wet weight C8 Acid PFOA Amount Recovered (ng/g) wet weight NR NR NR NR 1.21 3.80 1.21 303 0.733 0.733 NR NR 3.72 295 NR NR NR NR NR NR 1.20 4.14 1.20 343 5.04 5.04 9.20 294 ND 3.93 ND 328 6.47 6.47 6.22 6.22 11.7 361 10.6 387 Recovery (% ) NR NR 65 75 75 74 NR NR NR NR 74 85 104 72 98 82 131 89 110 95 'Sample residue exceeds the spiking level significantly (3x spiking level); therefore, an accurate recovery value cannot be calculated. ND = Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR = Not Reported due to Quality Control failures, see Table V for re-extract data. 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 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table IV. Matrix Spike Recovery of PFOA in Soil Samples Continued Sample Description D A L -S B -605R -0-0005 (C0150460 Spk K, 4 ppto Spilw) D A L -S B -605R -0-0005 (C0159460 Spk L, 400 ppb S pite) D A L-S B -605R -0-0010 (C0159401 Spk C, 4 ppb S pite) D A L-S B -605R -0-0010 (C01SS4S1 Spk D, 400 ppb Spike) D A L-S B -605R -0-0050 (C01SS4S2 Spk E, 4 ppb Spike) DAL-S B -605R -0 -005 0 (C0159462 Spk F, 400 ppb S pite) D A L-S B -605R -0-0100 (C0150463 Spk Q, 4 ppb S pite) D A L -S B -605R -0-0100 (C0150403 Spk H, 400 ppb Spike) D A L-S B -138R -0-0000 (C0150404 Spk 1,4 ppb Spike) D A L-S B -138R -0-0000 (C0150404 Spk J. 400 ppb S pite) D A L-S B -138R -0-0005 (C01504M Spk K, 4 ppb Spike) D A L-S B -138R -0-0005 (C0150400 Spk L, 400 ppb S pite) D A L-S B -138R-0-0010 (C0150407 Spk C, 4 ppb S pite) D A L -S B -138R -0-0010 (C0150407 Spk D, 400 ppb S pite) D A L -S B -1 38R -0 -00 50 (C0159400 Spk E, 4 ppb S pite) D A L -S B -1 38R -0 -00 50 (C0150405 Spk F, 400 ppb Spike) D A L -S B -1 38R -0 -010 0 (C0150470 Spk G, 4 ppb Spike) D A L -S B -138R -0-0100 (C0150470 Spk H, 400 ppb S pite) D A L-S B -138R -D B -0100 (C0159471 Spk 1,4 ppb S pite) DAL-SB-138R-DB-0100 (C0159471 Spk J, 400 ppb S pite) D F 0 5-S S -S S 0 3-0-000 0 (C0100071 Spk G, 4 ppb S pite) D F 0 5-S S -S S 0 3-0-000 0 (C0100071 Spk H, 400 ppb Spike) Amount Spiked (ng/g) 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 Amt Found in Sample (ng/g) wet weight Cfl Acid PFOA Amount Recovered (ng/g) wet weight 6.63 6.63 10.5 326 2.17 2.17 6.00 281 0.808 0.808 3.99 369 0.627 0.627 3.67 286 23.8 23.8 29.0 366 42.9 42.9 48.7 407 35.0 35.0 46.7 337 NR NR NR NR 278 345 278 546 247 378 247 543 NR NR NR NR Recovery (% ) 97 80 96 70 80 92 76 71 86 91 76 NR NR * 67 74 NR NR `Sample residue exceeds the spiking level significantly (3x spiking level); therefore, an accurate recovery value cannot be calculated. ND - Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR = Not Reported due to Quality Control failures, see Table V for re-extract data. Note: Since this summary table shows rounded results, recovery values may vary slightly from the values In the raw data. Exygen Research Page 40 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table IV. Matrix Spike Recovery of PFOA in Soil Samples Continued Sample Description D F 0 5-S S -S S 0 3-0-000 3 (00100872 Spk J, 4 ppb SpJk) D F05-S S -S S 03-0-0003 (C0160872 Spk J, 400 ppb Spik) D F 05-S S -S S 04-0-0000 (C0160S73 Spk K, 4 ppb Splks) D F 0 5-S S -S S 0 4-0-000 0 (CO10O873 Spk L, 400 ppb Splk) D F 0 5-S S -S S 0 4-0-000 3 (C016Q874 Spk C, 4 ppb Spika) D F 0 5-S S -S S 0 4-0-000 3 (CO16O074 Spk D, 400 ppb Splk) D F 0 5-S S -S S 05-Q -0000 (C0100879 Spk E, 4 ppb Splk) D F 0 5-S S -S S 0 5-0-000 0 (CO10O075 Spk F, 400 ppb Splk) D F 0 5 -S S -S S 0 5 -0 -0 0 0 3 (C01S087S Spk 6 ,4 ppb Spika) D F 0 5-S S -S S 0 5-0-000 3 (00100870 Spk H, 400 ppb Splk) D F05-S S -S S 0 5-D B -0 003 (C01S0877 Spk 1,4 ppb Spika) D F05-S S -S S 0 5-D B -0 003 (CO10O877 Spk J, 400 ppb Splk) D F 0 9 -S S -0 0 0 6 -0 -0 0 0 0 (CO10O078 Spk K, 4 ppb Splk) D F 0 9 -S S -0 0 0 6 -0 -0 0 0 0 (CO10O878 Spk t , 400 ppb Splk) D F 0 9 -S S -0 0 0 6 -0 -0 0 0 3 (CO10O879 Spk C, 4 ppb Splk) D F 0 9 -S S -0 0 0 6 -0 -0 0 0 3 (C01S0879 Spk D, 400 ppb Spika) D F 0 9-S S -000 6-0 -00 10 (COIOHO Spk E. 4 ppb Spika) D F 0 9 -S S -0 0 0 6 -0 -0 0 1 0 (C0100800 Spk F, 400 ppb Splk) D F 0 9-S S -000 7-0 -00 00 (C0180881 Spk G, 4 ppb Splk) D F 0 9 -S S -0 0 0 7 -0 -0 0 0 0 (CO10O801 Spk H, 400 ppb Splk) Amount Spiked (ng/g) 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 Amt Found in Sample (ng/g) wet weight C8 Acid PFOA Amount Recovered (ng/g) wet weight NR NR NR NR NR NR NR NR 4.02 4.02 5.01 5.01 7.72 333 7.69 310 6.26 6.26 9.60 314 6.44 6.44 8.56 298 13.2 13.2 18.0 392 14.2 14.2 19.8 332 7.68 7.68 13.0 335 150 136 150 357 Recovery NR NR NR NR 93 82 67 76 84 77 53 73 95 * 79 133 82 52 'Sample residue exceeds the spiking level significantly (3x spiking level); therefore, an accurate recovery value cannot be calculated. ND = Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR = Not Reported due to Quality Control failures, see Table V for re-extract data. 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 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table IV. Matrix Spike Recovery of PFOA in Soil Samples Continued Sam ple Description D F 0 9 -S S -0 0 0 7 -0 -0 0 0 3 (C0160882 Spk 1,4 ppb Spike) D F 0 9 -S S -0 0 0 7 -0 -0 0 0 3 (C0160682 Spk J, 400 ppb Spike) D F 0 9-S S -000 7-0 -00 10 (C0180883 Spk K, 4 ppb Spike) D F 0 9-S S -000 7-0 -00 10 (C0160B83 Spk L, 400 ppb Spike) D F 0 9-S S -C 007 -D B -00 10 (C0160884 Spk C, 4 ppb Spike) D F 09-S S -0007-D B -0010 (C160884 Spk D, 400 ppb Spik) DF09-SS-0008-0-000 (C0160885 Spk E, 4 ppb Spike) D F 0 9-S S -000 8-0 -00 00 (C01S0885 Spk F, 400 ppb Spike) D F 0 9 -S S -0 0 0 8 -0 -0 0 0 3 (C01S08M Spk 6 ,4 ppb Spike) D F 0 9 -S S -0 0 0 8 -0 -0 0 0 3 (C0160886 Spk H, 400 ppb Spike) D F 0 9-S S -000 8-0 -00 10 (C0100M7 Spk 1,4 ppb Spike) D F 0 9 -S S -0 0 0 8 -0 -0 0 1 0 (C0160687 Spk J, 400 ppb Spike) D F 09-S S -0009-0-0000 (C0100888 Spk K, 4 ppb Spike) D F 0 9 -S S -0 0 0 9 -0 -0 0 0 0 (C0160888 Spk L, 400 ppb Spike) D F 0 9 -S S -0 0 0 9 -0 -0 0 0 3 (COfOOMO Spk C, 4 ppb Spike) D F 09-S S -0009-0-0003 (C0100M9 Spk D. 400 ppb Spike) D F0 9-S S -000 9-D B -0 003 (C0180890 Spk E, 4 ppb Spike) D F09-S S -000 9-D B -0 003 <00160800 Spk F, 400 ppb Spike) D F 0 9-S S -000 9-0 -00 10 (C0160801 Spk G, 4 ppb Spike) D F 0 9-S S -000 9-0 -00 10 (C0160881 Spk H, 400 ppb Spike) Amount Spiked (ng/g) 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 Amt Found in Sample (ng/g) wet weight ca Acid PFOA Amount Recovered (ng/g) wet weight 111 109 111 319 41.8 41.8 44.2 266 34.8 34.8 38.7 425 152 150 152 518 131 120 131 371 108 99.2 108 426 8.88 8.88 14.2 435 10.8 15.8 10.8 486 10.9 10.9 14.7 394 16.7 16.7 20.6 422 Recovery <%) * 52 56 * 98 92 60 80 133 107 125 119 95 96 101 `Sample residue exceeds the spiking level significantly (3x spiking level); therefore, an accurate recovery value cannot be calculated. NP = Not detected at ex above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR = Not Reported due to Quality Control bilures, see Table V for re-extract data. Note: Since this summary table shows rounded results, recovery values may vary slightly from the values In the raw data. Exygen Research Page 42 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table IV. Matrix Spike Recovery of PFOA in Soil Samples Continued Sample Description D F 0 9-S S -0010-0-0000 (C0190892 Spk 1.4 ppb Spika) D F 0 9-S S -0010 -0-00 00 (C01S0892 Spk J, 400 ppb Spika) D F 0 9-S S -0010 -0-00 03 (C01S0893 Spk K. 4 ppb Spika) DF09-SS-001 0-0-0003 (C0190893 Spk L, 400 ppb Spika) D F 0 9 -S S -0 0 1 0 -0 -0 0 1 0 (C0190894 Spk C, 4 ppb Spika) D F 0 9-S S -0010 -0-00 10 (C0190994 Spk D, 400 ppb Spika) D F09-SS-0011-0-0000 (C0180895 Spk E, 4 ppb Spika) D F 0 9 -S S -0 0 1 1-0-0000 (C0180995 Spk F, 400 ppb Spika) D F 0 9 -S S -0 0 1 1-0-0003 (C01908H Spk G, 4 ppb Spika) D F09-SS-0011-0-0003 (C0180898 Spk H, 400 ppb Spika) D F09-SS-0011-0-0010 (C0180S97 Spk 1,4 ppb Spika) D F 0 9 -S S -0 0 1 1-0-0010 (C0180897 Spk J, 400 ppb Spika) D F 0 9 -S S -0 0 1 2 -0 -0 0 0 0 (C0180898 Spk K, 4 ppb Spika) D F 0 9-S S -0012 -0-00 00 (coieoaea spk i , aoo ppb Spike) D F 0 9-S S -001 2-D B -0 000 (C0180899 Spk C, 4 ppb Spika) D F 0 9-S S -001 2-D B -0 000 (C0180899 Spk 0 ,400 ppb Spika) D F 0 9-S S -0012-0-00 03 (C0180900 Spk E, 4 ppb Spika) D F 0 9-S S -0012-0-00 03 (C0180900 Spk F, 400 ppb Spika) D F 0 9-S S -0012-0-00 10 (C0160901 Spk G, 4 ppb Spika) D F 0 9 -S S -0 0 1 2 -0 -0 0 1 0 (C0180901 Spk H, 400 ppb Spika) Amount Spiked (ng/g) 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 4 400 Amt Found in Sample (ng/g) wet weight C8 Add PFOA Amount Recovered (ng/g) wet weight 638 706 638 1070 386 367 386 816 353 440 353 683 222 234 222 507 247 307 247 550 NR NR NR NR NR NR NR NR 484 433 484 764 NR NR NR NR 418 373 418 692 Recovery (% ) * 108 108 83 71 * 76 NR NR NR NR * 70 NR NR 69 'Sample residue exceeds the spiking level significantly (3x spiking level); therefore, an accurate recovery value cannot be calculated. ND = Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR = Not Reported due to Quality Control bilures, see Table V for reextract data. 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 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table IV. Matrix Spike Recovery of PFOA in Soil Samples Continued Sam ple Description D F 0 9-S S -0013 -0-00 00 (C0160902 Spk 1,4 ppb Spike) D F 0 9 -S S -0 0 1 3-0 -0 0 0 0 (C0160902 Spk J, 400 ppb Spike) D F 0 9-S S -0013 -0-00 03 (C0140003 Spk K, 4 ppb Spike) D F 0 9-S S -0013 -0-00 03 (C0140903 Spk L, 400 ppb Spike) D F 0 9 -S S -0 0 1 3-0 -0 0 1 0 (C0160404 Spk C. 4 ppb Spike) D F 0 9 -S S -0 0 1 3-0 -0 0 1 0 (C0140004 Spk D. 400 ppb Spike) D A L-S B -602R -0-0000 (C0144247 Spk K. 4 ppb Spike) D A L -S B -602R -0-0000 (C0164247 Spk L, 400 ppb Spike) D A L -S B -602R -0-0005 (C0144244 Spk C, 4 ppb Spike) D A L -S B -602R -0-0005 (C0144244 Spk D, 400 ppb Spike) D A L -S B -6 02R -0 -00 10 (C0144249 Spk E, 4 ppb Spike) D A L -S B -6 02R -0 -00 10 (C0164240 Spk F, 400 ppb Spike) D A L-S B -602R -0-0050 (C01442S0 Spk G, 4 ppb Spike) D A L -S B -602R -0-0050 (C0144250 Spk H, 400 ppb Spike) D A L -S B -6 02R -0 -0100 (C01S42S1 Spk 1,4 ppb Spike) DA L -S B -6 02R -0-0100 (C01M251 Spk J, 400 ppb Spike) D A L -S B -6 01R -0 -00 00 (C0144252 Spk K, 4 ppb Spike) DAL-SB-601 R-0-0000 (C01442S2 Spk L, 400 ppb Spike) DAL-SB-601 R-DB-0000 (C0144253 Spk C, 4 ppb Spike) DAL-SB-601 R-DB-0000 (C0164253 Spk D, 400 ppb Spike) Amount Spiked (ng/g) Amt Found In Sample (ng/g) wet weight C8 Add PFOA Amount Recovered (ng/g) w et weight 4 18.5 400 . 18.5 23.8 342 4 22.2 400 22.2 23.7 377 4 43.3 400 43.3 46.7 289 4 4.75 400 4.75 7.58 305 4 5.69 400 5.69 8.80 327 4 3.39 400 3.39 6.38 415 4 0.630 400 0.630 3.41 304 4 0.555 400 0.555 3.00 277 4 1.57 400 1.57 4.51 383 4 1.95 5.52 400 1.95 494 Recovery (% ) * 81 * 89 * 61 71 75 78 80 75 103 70 76 61 69 74 95 89 123 `Sample residue exceeds the spiking level significantly (3x spiking level); therefore, an accurate recovery value cannot be calculated. ND - Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR = Not Reported due to Quality Control failures, see Table V for re-extract data. 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 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table IV. Matrix Spike Recovery of PFOA in Soil Samples Continued Sample Description D A L-S B -601R -0-0005 (C01S4254Spk E, 4 ppb Spike) DAL-SB-601 R-0-0005 (C0164254 Spk F, 400 ppb Spike) DAL-SB-601 R-0-0010 (C0164255Spk 0 ,4 ppb Spikt) DAL-SB-601 R-0-0010 (C01I42S5 Spk H, 400 ppb Spike) DAL-SB-601 R-DB-0010 (C0164256 Spk t, 4 ppb Spilw) DAL-SB-601 R-DB-0010 (C0164256 Spk J, 400 ppb Spikt) DAL-SB-601 R-0-0050 (C0104257 Spk K, 4 ppb Splka) DAL-SB-601 R-0-0050 (C0104257 Spk L, 400 ppb Spike) DAL-SB-601 R-0-0100 (C0104259 Spk C, 4 ppb Spike) DAL-SB-601 R-0-0100 (C0164259 Spk D, 400 ppb Splk) DAL-SB-601 R-DB-0100 (C01042S0 Spk E, 4 ppb Spike) DAL-SB-601 R-DB-0100 (C0164200 Spk F, 400 ppb Spik) Amount Spiked (n fl/a ) 4 400 4 400 4 400 4 400 4 400 4 400 Amt Found in Sample (ng/g) wet weight C8 Acid PFOA Amount Recovered (ng/g) wet weight 1.70 1.70 NR NR 1.90 1.90 0.728 0.728 0.414 0.414 0.406 0.406 5.15 394 NR NR 7.43 490 3.80 270 3.76 366 3.22 326 Recovery (% ) 86 98 NR NR 138 122 77 67 84 91 70 81 Average: Standard Deviation: 80 19 'Sample residue exceeds the spiking level significantly (3x spiking level); therefore, an accurate recovery value cannot be calculated. ND = Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR = Not Reported due to Quality Control failures, see Table V for re-extract data. Note: Since this summary table shows rounded results, recovery values may vary slightly from the values In the raw data. Exygen Research Page 45 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table V. Matrix Spike Recovery of PFOA in Re-extracted Soil Samples Sample Description DF 13-S S -S S 03-0-00 00 (C0156344 Spk E, 400 ppb Splke) DF 13-S S -S S 03-0-00 03 (C0156345 Spk G, 400 ppb Splke) D F 1 3-S S -S S 0 5-0-000 0 (C01SSJ51 Spk M, 2000 ppb Splke) D F 1 3-S S -S S 05-0 -0 0 0 3 (C01563S2 Spk E, 2000 ppb Splke) D F 1 3-S S -S S 0 5-D B -0 003 (C01S6353Spk G, 2000 ppb Splke) DF 10-SS-SS01-0-0000 (C0156354 Spk 1,400 ppb Splke) D F 1 1-S S -S S 0 1-0 -0 0 0 3 (C0156367 Spk N, 400 ppb Splke) D F 1 1-S S -S S 0 2 -0 -0 0 0 0 (COI56366 Spk E, 2000 ppb Splke) D F 11-S S -S S 0 2 -0 -0 0 0 3 (C0156370 Spk G, 600 ppb Splke) DF 11-SS-SS03-0-0000 (C0156371 Spk 1,400 ppb Splke) DF 11-SS-SSO3-0-OOO3 (COI56373 Spk M, 400 ppb Splke) D F 0 5-S S -S S 0 2-0-000 3 (C0156361 Spk 1,400 ppb Splke) DF 12-SS-SSO1-0-0003 (C0156383 Spk M, 400 ppb Splke) D F 1 2-S S -S S 0 2-0-000 0 (C0156384 Spk E, 2000 ppb Splke) D F 1 2-S S -S S 0 2-0-000 3 (C0156385 Spk G, 4000 ppb Splke) DF 12-S S -S S 03-0-0000 (C0156386 Spk 1,40 ppb Splke) DF 12-SS-SS04-0-0000 (C0156368 Spk M, 6000 ppb Splke) D F 1 2-S S -S S 0 4-0-000 3 (C0156369 Spk E, 20,000 ppb Splke) DF13-SS-SS01 -0-0003 (COI56363 Spk M, 400 ppb Splke) D F 1 3 -S S -S S 0 2 -0 -0 0 0 0 (COI56304 Spk E, 4000 ppb Splke) Amount Spiked (ng/g) 400 400 2000 2000 2000 400 400 2000 800 400 400 400 400 2000 4000 40 8000 20000 400 4000 Amt Found in Sample (ng/g) wet weight C8 Acid PFOA Amount Recovered (ng/g) wet weight 344 422 880 864 714 NR 72.7 NR 579 81.6 NR 282 585 824 2120 3180 1750 NR 402 NR 1140 357 NR 510 160 1470 2880 23.5 3260 9280 450 2540 481 2520 5400 45.7 10600 29800 794 4760 Recovery (% ) 60 101 62 116 52 NR 82 NR 70 69 NR 57 80 53 63 56 92 103 86 56 NR = Not reported due to quality control failures. Note: Since this summary table shows rounded results, recovery values may vary slightly from the values In the raw data. i Exygen Research Page 46 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table V. Matrix Spike Recovery of PFOA in Re-extracted Soil Samples Continued Sam ple Description D F13-SS-SS02-0-0003 (C0156395 Spk G, 4000 ppb Spike) D A L -S B -6 03R -0 -00 00 (C0159446 Spk E, 4 ppb Spike) D A L -S B -6 03R -0 -00 50 (C0159449 Spk M, 4 ppb Spike) D A L -S B -6 04R -0 -00 00 (C0150453 Spk 1,4 ppb Spike) D A L -S B -6 04R -0 -00 05 (C0150454 Spk K, 4 ppb Spike) D A L -S B -1 38R -0 -00 50 (C0159468 Spk G, 400 ppb Spike) D F 0 5-S S -S S 0 3 -0 -0 0 0 0 (00100871 Spk 1,400 ppb Spike) D F05-SS-SS03-0-0003 (C0100872 Spk K, 400 ppb Spike) D F05-SS-SS04-0-0000 (C0180873 Spk M, 4 ppb Spike) D F09-SS-0011-0-0010 (00160807 Spk K, 400 ppb Spike) D F09-SS-0012-0-0000 (C0160808 Spk M, 400 ppb Spike) D F09-SS-0012-0-0003 (00160900 Spk G, 400 ppb Spike) D A L -S B -6 01R -0 -00 10 (C01642S5 Spk I, 4 ppb Spike) Amount Spiked (ng/g) 4000 4 4 4 4 400 400 400 4 400 400 400 4 Amt Found in Sample (ng/g) wet weight C8 Acid PFOA Amount Recovered (ng/g) wet weight 4760 NR 3.07 2.05 3.08 85.6 183 NR NR 205 NR NR NR 7920 NR 6.50 4.29 6.26 498 442 NR NR 455 NR NR NR Recovery (% ) 79 NR 86 56 80 103 65 NR NR 63 NR NR NR Avarag: Standard Davlation: 74 18 NR = Not reported due to quality control failures. Note: Since this summary table shows rounded results, recovery values may vary slightly from the values In the raw data. Exygen Research Page 47 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table VI. Matrix Spike Recovery of PFOA in Associated Trip Blanks S a m p le D e s c rip tio n D F 1 3 -S S -T R IP 1 -L S -0 6 0 2 2 1 (C 01 5 6 34 9 ,1.0 ppb Field Spike) D F 1 3 -S S -T R IP 1 -H S -0 6 0 2 2 1 (C 0156350,100 ppb Field Spike) D A L -S B -T R IP 1 -L S -0 6 0 3 0 2 (C0159444,1.0 ppb Field Spike) D A L -S B -T R IP 1 -H S -0 6 0 3 0 2 (C 0159445,100 ppb Field Spike) D F 0 5 -S S -T R IP 2 -L S -0 6 0 3 0 8 (C 0160868,1.0 ppb Field Spike) D F 0 5 -S S -T R IP 2 -H S -0 6 0 3 0 8 (C 0160869,100 ppb Field Spike) D A L -S B -T R IP -L S -0 6 0 3 1 6 (C 01 6 4 22 3 ,1.0 ppb Field Spike) D A L -S B -T R IP -H S -0 6 0 3 1 6 (C 0164224,10 ppb Field Spike) Am ount S p ik e d (n g /L ) Am t Found in S a m p le (n g /L ) C 8 A c id P F O A Am ount R e co ve re d (n g /L ) R e co ve ry (% ) 1000 100000 ND ND 989 96700 99 97 1000 100000 ND ND 862 81500 86 82 1000 100000 ND ND 985 96900 99 97 1000 10000 ND ND 907 8390 91 84 A verage: S ta n d a rd D e v ia tio n : ND = N ot detected a t o r above the L im it o f Q uantitation (LO Q ) o f 25 ng/L. Note: Since this summary table shows rounded results, recovery values may vary slightly from the values In the raw data. 92 .7 Exygen Research Page 48 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table VII. Surrogate Spike Recovery of 13C PFOA in Soil Samples Exygen ID Sam ple D escription C0156344 C0156344 Rep C0156344 Spk C C0156344 Spk D C0156345 C 0156345 Rep C0156345 Spk E C0156345 Spk F C0156346 C0156346 Rep C0156346 Spk G C0156346 Spk H C0156347 C0156347 Rep C0156347 Spk I C0156347 Spk J C 0 1 56351 C0156351 Rep C0156351 Spk K C0156351 Spk L C0156352 C0156352 Rep C0156352 Spk C C0156352 Spk D C0156353 C0156353 Rep C0156353 Spk E C0156353 Spk F C0156354 C0156354 Rep C0156354 Spk G C0156354 Spk H C0156355 C0156355 Rep C0156355 Spk 1 C0156355 Spk J C0156356 C0156356 Rep C0156356 Spk K C0156356 Spk L DF 13-S S -S S 03-0-0000 DF 13-S S -S S 03-0-0000 DF 13-S S -S S 03-0-0000 DF 13-S S -S S 03-0-0000 DF 13-S S -S S 03-0-0003 DF 13-S S -S S 03-0-0003 DF 13-S S -S S 03-0-0003 DF 13-S S -S S 03-0-0003 DF 13-S S -S S 04-0-0000 D F13-SS-SS04-0-0000 DF 13-S S -S S 04-0-0000 DF 13-S S -S S 04-0-0000 D F13-SS-SS04-0-0003 D F13-SS-SS04-0-0003 D F13-SS-SS04-0-0003 D F13-SS-SS04-0-0003 D F13-SS-SS05-0-0000 D F13-SS-SS05-0-0000 D F13-SS-SS05-0-0000 D F13-SS-SS05-0-0000 D F13-SS-SS05-0-0003 D F13-SS-SS05-0-0003 DF 13-S S -S S 05-0-0003 DF 13-S S -S S 05-0-0003 D F13-SS-SS05-D B-0003 DF 13-SS-SS 05-D B-0003 DF 13-SS-SS 05-D B-0003 DF 13-SS-SS 05-D B-0003 D F10-SS-SS01-0-0000 DF 10-S S -S S 01-0-0000 D F 1 0 -S S -S S 0 1 -0 -0 0 0 0 DF 10-S S -S S 01-0-0000 D F10-SS-SS01-0-0003 D F10-SS-SS01-0-0003 D F 1 0 -S S -S S 0 1 -0 -0 0 0 3 DF 10-S S -S S 01-0-0003 D F10-SS-SS02-0-0000 D F10-SS-SS02-0-0000 D F10-SS-SS02-0-0000 D F10-SS-SS02-0-0000 Am ount S p ik e d (ng /g ) 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 13c - p f o a Am ount R ecovered W e t W e ig h t (ng /g ) NR NR NR NR NR NR NR NR 4 .8 7 4 .7 8 2 .2 8 240 4 .1 8 4 .0 3 2 .6 9 267 NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR 3 .9 8 3.91 1.90 233 4 .4 9 4 .3 2 2 .1 0 246 R ecovery (% ) NR NR NR NR NR NR NR NR 122 120 57 60 105 101 67 67 NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR 100 98 48 58 112 108 53 62 Note: Since this sum m ary table shows rounded results, recovery values m ay vary slightly from the values In the raw data. NR = Not Reported due to Quality Control failures, see Table VIII for re-extract data. Exygen Research Page 49 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table VII. Surrogate Spike Recovery of 13C PFOA in Soil Samples Continued Exygen 10 Sam ple D escription C0156357 C 0156357 Rep C0156357 Spk C C0156357SpkD C0156358 C0156358 Rep C 0156358 Spk E C0156358 Spk F C0156360 C0156360 Rep C0156360 Spk G C0156360 Spk H C 0 156361 C0156361 Rep C0156361 S p kI C0156361 Spk J C0156362 C0156362 Rep CO156362 Spk K C0156362 Spk L C0156363 C 0156363 Rep C0156363 Spk C C0156363 Spk D C0156364 C0156364 Rep C 0156364 Spk E C0156364 Spk F C0156365 C0156365 Rep C0156365 Spk G C0156365 S pkH C0156366 C0156366 Rep C0156366 Spk I C0156366 S pkJ C0156367 C0156367 Rep C 0156367 Spk K C0156367 Spk L D F10-SS-SS02-D B-0000 DF 10-S S -S S 02-D B -0000 D F10-SS-SS02-D B-0000 D F10-SS-SS02-D B-0000 D F10-SS-SS02-0-0003 D F10-SS-SS02-0-0003 D F10-SS-SS02-0-0003 DF 10-S S -S S 02-0-0003 D F10-SS-SS03-0-0000 DF 10-S S -S S 03-0-0000 DF 10-S S -S S 03-0-0000 DF 10-SS-SS 03-0-0000 DF 10-S S -S S 03-0-0003 D F10-SS-SS03-0-0003 D F10-SS-SS03-0-0003 DF 10-S S -S S 03-0-0003 D F10-SS-SS04-0-0000 DF 10-S S -S S 04-0-0000 DF 10-S S -S S 04-0-0000 DF 10-S S -S S 04-0-0000 D F10-SS-SS04-0-0003 D F10-SS-SS04-0-0003 D F10-SS-SS04-0-0003 DF 10-S S -S S 04-0-0003 DF 10-S S -S S 05-0-0000 D F10-SS-SS05-0-0000 D F10-SS-SS05-0-0000 DF 10-S S -S S 05-0-0000 DF 10-S S -S S 05-0-0003 D F10-SS-SS05-0-0003 D F 1 0 -S S -S S 0 5 -0 -0 0 0 3 D F 1 0 -S S -S S 0 5 -0 -0 0 0 3 DF 11-S S -S S 01-0-0000 DF 11-S S -S S 01-0-0000 DF 11-S S -S S 01-0-0000 D F11-S S -S S 01-0-0000 D F11-S S -S S 01-0-0003 DF 11-SS-SS 01-0-0003 D F 1 1 -S S -S S 0 1 -0 -0 0 0 3 D F11-SS-SS01-0-0003 Am ount S p ik e d (n g /g ) 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 13c -p f o a Am ount R ecovered W e t W eig ht (n g /g ) 4 .9 0 4 .9 5 2 .4 3 250 4 .8 5 4 .9 8 2 .7 4 247 5.31 5 .2 5 2 .6 4 235 5 .6 2 5 .4 6 2 .9 0 270 5.41 5 .1 7 3 .1 5 253 3 .1 0 2 .7 8 1.78 258 3 .5 3 3 .2 8 1.78 243 2 .8 6 2 .7 8 1.38 235 2 .2 4 2 .9 0 1.61 250 NR NR NR NR R ecovery (% ) 123 124 61 63 121 125 69 62 133 131 66 59 141 137 73 68 135 129 79 63 78 70 45 65 88 82 45 61 72 70 35 59 56 73 40 63 NR NR NR NR Note: Since this sum m ary table shows rounded results, recovery values m ay vary slightly from the values in th e raw data. N R = Not Reported due to Quality Control failures, see Table V III for re-extract data. Exygen Research Page 50 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table VII. Surrogate Spike Recovery of 13C PFOA in Soil Samples Continued Exygen ID Sam ple D escription C0156368 C 0156368 Rep C0156368 Spk C C0156368 Spk D C0156370 CO156370 Rep C0156370 Spk E C0156370 Spk F C 0 1 56371 C0156371 Rep C0156371 Spk G C0156371 Spk H C0156372 CO156372 Rep C0156372 Spk I C0156372 Spk J C0156373 C0156373 Rep C0156373 Spk K C0156373 Spk L C0156374 C0156374 Rep C0156374 Spk C C0156374 Spk D C0156375 C0156375 Rep C0156375 Spk E C0156375 Spk F C0156376 C0156376 Rep C0156376 Spk G C0156376 Spk H C0156377 CO156377 Rep C0156377 Spk I C0156377 Spk J C0156378 C0156378 Rep C0156378 Spk K C0156378 Spk L D F11-SS-SS02-0-0000 D F11-SS-SS02-0-0000 D F11-SS-SS02-0-0000 D F11-SS-SS02-0-0000 D F11-SS-SS02-0-0003 D F 11-S S -S S 02-0-0003 D F11-SS-SS02-0-0003 D F11-SS-SS02-0-0003 D F11-SS-SS03-0-0000 D F11-SS-SS03-0-0000 D F11-SS-SS03-0-0000 D F11-SS-SS03-0-0000 D F11-SS-SS03-D B-0000 D F11-SS-SS03-D B-0000 D F11-SS-SS03-D B-0000 D F 11-S S -S S 03-D B -0000 D F11-SS-SS03-0-0003 DF 11-SS-SS 03-0-0003 DF 11-S S -S S 03-0-0003 D F11-SS-SS03-0-0003 D F11-SS-SS04-0-0000 D F11-SS-SS04-0-0000 D F11-SS-SS04-0-0000 D F11-SS-SS04-0-0000 D F11-SS-SS04-0-0003 D F 11-S S -S S 04-0-0003 DF 11-S S -S S 04-0-0003 D F 11-S S -S S 04-0-0003 DF 11-SS-SS 05-0-0000 D F 1 1 -S S -S S 0 5 -0 -0 0 0 0 D F 1 1 -S S -S S 0 5 -0 -0 0 0 0 D F 1 1 -S S -S S 0 5 -0 -0 0 0 0 D F 1 1 -S S -S S 0 5 -0 -0 0 0 3 D F 1 1 -S S -S S 0 5 -0 -0 0 0 3 D F 11-S S -S S 05-0-0003 D F 1 1 -S S -S S 0 5 -0 -0 0 0 3 D F05-SS-SS01-0-0000 D F05-SS-SS01-0-0000 D F05-SS-SS01-0-0000 D F05-SS-SS01-0-0000 Am ount S piked (n g /g ) 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 13c - p f o a Am ount R ecovered W e t W eig ht (ng /g ) NR NR NR NR NR NR NR NR NR NR NR NR 3 .7 4 3 .1 0 1.65 203 NR NR NR NR 4 .8 0 4 .5 2 2 .2 9 259 4 .1 5 4 .1 5 1.77 267 4 .6 6 4 .2 7 2 .5 5 288 4 .3 8 4 .1 2 2 .2 2 270 3 .7 4 3 .4 2 1.86 244 R ecovery (% ) NR NR NR NR NR NR NR NR NR NR NR NR 94 78 41 51 NR NR NR NR 120 113 57 65 104 104 44 67 117 107 64 72 110 103 56 68 94 86 47 61 Note: Since this sum m ary table shows rounded results, recovery values m ay vary slightly from the values in th e raw data. N R - Not Reported due to Quality Control failures, see Table V III for re-extract data. Exygen Research Page 51 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table VII. Surrogate Spike Recovery of 13C PFOA in Soil Samples Continued Exygen ID C0156379 C0156379 Rep C0156379 Spk C C0156379 Spk D C0156380 C0156380 Rep C0156380 Spk E C0156380 Spk F C 0156381 C0156381 Rep C0156381 Spk G C0156381 Spk H C0156382 C0156382 Rep C0156382 Spk I C0156382 Spk J C0156383 C 0156383 Rep C0156383 Spk K C 0156383 Spk L C0156384 C0156384 Rep C 0156384 Spk C C 0156384 Spk D C0156385 C 0156385 Rep C0156385 Spk E C0156385 Spk F C0156386 C0156386 Rep C0156386 Spk G C0156386 Spk H C0156387 C0156387 Rep C0156387 Spk I C0156387 Spk J C0156388 C0156388 Rep C 0156388 Spk K C0156388 Spk L Sam ple D escription D F05-SS-SS01-0-0003 D F05-SS-SS01-0-0003 D F05-SS-SS01-0-0003 D F05-SS-SS01-0-0003 D F05-SS-SS02-0-0000 D F05-SS-SS02-0-0000 D F05-SS-SS02-0-0000 D F05-SS-SS02-0-0000 D F05-SS-SS02-0-0003 D F05-SS-SS02-0-0003 D F05-SS-SS02-0-0003 D F05-SS-SS02-0-0003 D F 12-S S -S S 01-0-0000 D F 12-S S -S S 01-0-0000 DF 12-S S -S S 01-0-0000 DF 12-S S -S S 01-0-0000 DF 12-S S -S S 01-0-0003 DF 12-S S -S S 01-0-0003 D F12-SS-SS01-0-0003 DF 12-S S -S S 01-0-0003 DF 12-S S -S S 02-0-0000 D F12-SS-SS02-0-0000 D F12-SS-SS02-0-0000 D F12-SS-SS02-0-0000 D F12-SS-SS02-0-0003 DF 12-S S -S S 02-0-0003 DF 12-S S -S S 02-0-0003 D F 1 2 -S S -S S 0 2 -0 -0 0 0 3 DF 12-S S -S S 03-0-0000 DF 12-S S -S S 03-0-0000 DF 12-S S -S S 03-0-0000 DF 12-S S -S S 03-0-0000 D F12-SS-SS03-0-0003 D F12-SS-SS03-0-0003 DF 12-S S -S S 03-0-0003 DF 12-S S -S S 03-0-0003 DF 12-S S -S S 04-0-0000 D F12-SS-SS04-0-0000 DF 12-S S -S S 04-0-0000 DF 12-S S -S S 04-0-0000 Am ount S p ik e d (n g /g ) 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 13c -p f o a Am ount R ecovered W e t W eig ht (ng /g ) 4 .7 0 4 .5 9 3 .0 4 278 1.37 1.16 0 .6 2 5 248 NR NR NR NR 3 .8 7 2 .6 6 1.90 270 NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR 4 .5 4 4 .2 9 2 .3 2 2 .1 4 NR NR NR NR R ecovery (% ) 118 115 76 70 34 29 16 62 NR NR NR NR 97 67 48 68 NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR 114 107 58 1 NR NR NR NR Note: Since this sum m ary table shows rounded results, recovery values m ay vary slightly from the values in the raw data. N R = Not Reported due to Quality Control failures, see Table V III for re-extract data. Exygen Research Page 52 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table VII. Surrogate Spike Recovery of 13C PFOA in Soil Samples Continued Exygen 10 C0156389 C0156389 Rep C0156389 S pkC C0156389 Spk D C0156390 C 0156390 Rep C0156390 Spk E C0156390 Spk F C 0156391 C0156391 Rep C0156391 Spk G C0156391 Spk H C0156392 C0156392 Rep C0156392 Spk I C0156392 Spk J C0156393 C0156393 Rep C 0156393 Spk K C0156393 Spk L C0156394 CO156394 Rep C0156394 S pkC C0156394 Spk D C0156395 C0156395 Rep C0156395 Spk E C0156395 Spk F C0159446 C0159446 Rep C0159446 Spk E C0159446 Spk F C0159447 C0159447 Rep C0159447 S pkG C0159447 Spk H C0159448 C0159448 Rep C0159448 Spk I C0159448 Spk J Sam ple D escription D F12-SS-SS04-0-0003 DF 12-S S -S S 04-0-0003 DF 12-S S -S S 04-0-0003 DF 12-S S -S S 04-0-0003 DF 12-S S -S S 05-0-0000 DF 12-S S -S S 05-0-0000 DF 12-S S -S S 05-0-0000 DF 12-S S -S S 05-0-0000 D F12-SS-SS05-0-0003 D F12-SS-SS05-0-0003 D F12-SS-SS05-0-0003 D F12-SS-SS05-0-0003 DF 13-S S -S S 01-0-0000 D F13-SS-SS01-0-0000 DF 13-S S -S S 01-0-0000 DF 13-S S -S S 01-0-0000 D F13-SS-SS01-0-0003 DF 13-S S -S S 01-0-0003 DF 13-S S -S S 01-0-0003 D F 1 3 -S S -S S 0 1 -0 -0 0 0 3 DF 13-S S -S S 02-0-0000 DF 13-S S -S S 02-0-0000 DF 13-S S -S S 02-0-0000 DF 13-S S -S S 02-0-0000 D F13-SS-SS02-0-0003 D F13-SS-SS02-0-0003 DF 13-S S -S S 02-0-0003 DF 13-S S -S S 02-0-0003 D AL-SB-603R -0-0000 D AL-SB-603R -0-0000 DAL-SB-603R -0-0000 DAL-SB-603R -0-0000 DAL-SB-603R -0-0005 DAL-SB-603R -0-0005 D AL-SB-603R -0-0005 DAL-SB-603R -0-0005 D A L-S B -603R -0-0010 D AL-SB-603R -0-0010 D A L-S B -603R -0-0010 D AL-SB-603R -0-0010 Am ount S piked (ng /g ) 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 13c -p f o a Am ount R ecovered W et W eig ht (ng /g ) NR NR NR NR 2 .3 4 2 .3 0 1.57 335 1.71 1.70 0 .9 4 4 281 0 .8 8 0 1.37 0 .5 4 7 216 NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR 4 .6 2 4 .3 2 2 .5 3 265 4.4 7 4 .8 6 2 .8 2 285 R ecovery (% ) NR NR NR NR 59 58 39 84 43 43 24 70 22 34 14 54 NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR 116 108 63 66 112 122 71 71 Note: Since this sum m ary table shows rounded results, recovery values m ay vary slightly from the values in the raw data. NR n Not Reported due to Quality Control failures, see Table VIII for re-extract data. Exygen Research Page 53 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table VII. Surrogate Spike Recovery of 13C PFOA in Soil Samples Continued Exygen ID Sam ple D escription C0159449 C0159449 Rep C0159449 Spk K C0159449 Spk L C0159450 CO159450 Rep C0159450 Spk C C0159450 Spk D C 0 159451 C0159451 Rep C0159451 Spk E C0159451 Spk F C0159453 C0159453 Rep C 0159453 Spk G C0159453 Spk H C0159454 C0159454 Rep C0159454 Spk I C0159454 Spk J C0159455 CO159455 Rep C 0159455 Spk K C0159455 Spk L C0159456 C0159456 Rep C0159456 Spk C C0159456 Spk D C0159457 C0159457 Rep C0159457 Spk E C0159457 Spk F C0159458 C0159458 Rep C0159458 Spk G C0159458 Spk H C0159459 CO159459 Rep C0159459 Spk I C0159459 Spk J D AL-SB-603R -0-0050 D AL-SB-603R -0-0050 D AL-SB-603R -0-0050 D AL-SB-603R -0-0050 D A L-SB-603R -0-0100 D A L-SB-603R -0-0100 D A L-SB-603R -0-0100 D A L-SB-603R -0-0100 D A L-SB-603R -D B-0100 D A L-SB-603R -D B-0100 D AL-SB-603R -D B-0100 D AL-SB-603R -D B-0100 D AL-SB-604R -0-0000 D AL-SB-604R -0-0000 D AL-SB-604R -0-0000 D AL-SB-604R -0-0000 D AL-SB-604R -0-0005 D AL-SB-604R -0-0005 D AL-SB-604R -0-0005 D AL-SB-604R -0-0005 D A L-S B -604R -0-0010 D A L-S B -604R -0-0010 D AL-SB-604R -0-0010 D AL-SB-604R -0-0010 D AL-SB-604R -0-0050 D AL-SB-604R -0-0050 D AL-SB-604R -0-0050 D AL-SB-604R -0-0050 D AL-SB-604R -0-0100 D AL-SB-604R -0-0100 D AL-SB-604R -0-0100 D AL-SB-604R -0-0100 D AL-SB-605R -0-0000 D AL-SB-605R -0-0000 D AL-SB-605R -0-0000 D AL-SB-605R -0-0000 D AL-SB-605R -D B-0000 D AL-SB-605R -D B-0000 D AL-SB-605R -D B-0000 D AL-SB-605R -D B-0000 Am ount S p ik e d (n g /g ) 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 ,3c -p f o a Am ount R ecovered W e t W eig ht (ng /g ) NR NR NR NR 5 .2 4 5 .0 4 2 .5 6 296 4 .7 9 4 .8 5 2 .9 4 287 NR NR NR NR NR NR NR NR 5 .2 6 5 .3 2 3 .0 8 348 5 .1 6 5 .2 9 3 .3 4 305 5 .6 9 5 .1 4 3 .6 3 314 5 .1 8 5 .1 5 3 .1 4 351 4 .7 6 5 .2 2 2 .9 5 362 R ecovery (% ) NR NR NR NR 131 126 64 74 120 121 74 72 NR NR NR NR NR NR NR NR 132 133 77 87 129 132 84 76 142 129 91 79 130 129 79 88 119 131 74 91 Note: Since this sum mary table shows rounded results, recovery values m ay vary slightly from the values In th e raw data. NR = Not Reported due to Quality Control failures, see Table V III for reextract data. Exygen Research Page 54 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table VII. Surrogate Spike Recovery of 13C PFOA in Soil Samples Continued Exygen ID C0159460 C0159460 Rep C0159460 Spk K C0159460 Spk L C0159461 C0159461 Rep C0159461 Spk C C0159461 Spk D CO159462 C0159462 Rep CO159462Spk E C0159462 Spk F C0159463 CO159463 Rep C 0159463 Spk G C0159463 Spk H C0159464 C0159464 Rep C0159464 S pkI C0159464 Spk J C0159466 C0159466 Rep C0159466 Spk K C0159466 Spk L C0159467 C0159467 Rep C0159467S pkC C0159467 Spk D C0159468 C0159468 Rep C0159468 Spk E C0159468 Spk F C0159470 C0159470 Rep C0159470 Spk G C0159470 Spk H C 0 159471 C0159471 Rep C0159471 S pkI C0159471 S pkJ C0160871 C0160871 Rep C0160871 Spk G C0160871 Spk H S am ple D e scrip tio n D A L -S B -60 5R -0 -00 05 D A L -S B -60 5R -0 -00 05 D A L -S B -60 5R -0 -00 05 D A L -S B -60 5R -0 -00 05 D A L -S B -60 5R -0 -0010 D A L -S B -60 5R -0 -0010 D A L -S B -6 0 5 R -0 -0 0 10 D A L -S B -6 0 5 R -0 -0 0 10 D A L -S B -60 5R -0 -00 50 D A L -S B -6 0 5 R -0 -0 0 5 0 D A L -S B -60 5R -0 -00 50 D A L -S B -60 5R -0 -00 50 D A L -S B -60 5R -0 -01 00 D A L -S B -6 0 5 R -0 -0 1 00 D A L -S B -6 0 5 R -0 -0 1 00 D A L -S B -6 0 5 R -0 -0 1 00 D A L -S B -13 8R -0 -00 00 D A L -S B -13 8R -0 -00 00 D A L -S B -13 8R -0 -00 00 D A L -S B -13 8R -0 -00 00 D A L -S B -13 8R -0 -00 05 D A L -S B -13 8R -0 -00 05 D A L -S B -13 8R -0 -00 05 D A L -S B -13 8R -0 -00 05 D A L -S B -13 8R -0 -0010 D A L -S B -13 8R -0 -0010 D A L -S B -13 8R -0 -0010 D A L -S B -13 8R -0 -0010 D A L -S B -13 8R -0 -00 50 D A L -S B -13 8R -0 -00 50 D A L -S B -13 8R -0 -00 50 D A L -S B -13 8R -0 -00 50 D A L -S B -13 8R -0 -01 00 D A L -S B -1 3 8 R -0 -0 1 00 D A L -S B -13 8R -0 -01 00 D A L -S B -13 8R -0 -01 00 D A L -S B -13 8R -D B -0100 D A L -S B -13 8R -D B -0100 D A L -S B -13 8R -D B -0100 D A L -S B -13 8R -D B -0100 D F 0 5 -S S -S S 0 3 -0 -0 0 0 0 D F 0 5 -S S -S S 0 3 -0 -0 0 0 0 D F 0 5 -S S -S S 0 3 -0 -0 0 0 0 D F 0 5 -S S -S S 0 3 -0 -0 0 0 0 Am ount Spiked (ng/g) 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 ,3c - p f o a A m ount R e cove red W et W eight (ng/g) 5.60 5.37 2.59 323 5.94 5.89 2.71 278 5.54 5.53 2.98 357 5.46 5.06 2.98 276 5.45 5.52 2.93 329 4.82 4.85 2.67 359 5.58 5.35 3.03 291 NR NR NR NR 4.54 5.06 2.78 274 4.40 4.19 2.41 328 NR NR NR NR R e cove ry (% ) 140 134 65 81 149 147 68 70 139 138 75 89 137 127 75 69 136 138 73 82 121 121 67 90 140 134 76 73 NR NR NR NR 114 127 70 69 110 105 60 82 NR NR NR NR Note: Since this summary table shows rounded results, recovery values may vary slightly from the values in the raw data. NR = Not Reported due to Quality Control failures, see Table VIII for re-extract data. Exygen Research Page 55 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table VII. Surrogate Spike Recovery of 13C PFOA in Soil Samples Continued Exygen ID Sam ple D escription C 0160872 CO160872 Rep C 0160872 Spk I C0160872 Spk J CO160873 CO160873 Rep C 0160873 Spk K C0160873 Spk L CO160874 C0160874 Rep C0160874 Spk C C0160874 Spk D C0160875 C0160875 Rep C0160875 Spk E CO160875 Spk F C0160876 CO160876 Rep CO160876 S pk G C0160876 Spk H C0160877 C0160877 Rep C0160877 S p kI C0160877 Spk J C0160878 C0160878 Rep C 0160878 Spk K C0160878 Spk L CO160879 C0160879 Rep C0160879 Spk I C0160879 Spk J C 0160880 CO160880 Rep CO160880 Spk K C 0160880 Spk L C0160881 C0160881 Rep C0160881 Spk C C0160881 Spk D D F05-SS-SS03-0-0003 D F05-SS-SS03-0-0003 D F05-SS-SS03-0-0003 D F05-SS-SS03-0-0003 D F05-SS-SS04-0-0000 D F05-SS-SS04-0-0000 D F05-SS-SS04-0-0000 D F05-SS-SS04-0-0000 D F05-SS-SS04-0-0003 D F05-SS-SS04-0-0003 D F05-SS-SS04-0-0003 D F05-SS-SS04-0-0003 D F05-SS-SS05-0-0000 D F05-SS-SS05-0-0000 D F05-SS-SS05-0-0000 D F05-SS-SS05-0-0000 D F05-SS-SS05-0-0003 D F05-SS-SS05-0-0003 D F05-SS-SS05-0-0003 D F05-SS-SS05-0-0003 D F05-SS-SS05-D B-0003 D F05-SS-SS05-D B-0003 D F05-SS-SS05-D B-0003 D F05-SS-SS05-D B-0003 D F09-SS-0006-0-0000 D F09-S S -0006-0-0000 D F09-SS-0006-0-0000 D F09-SS-0006-0-0000 D F09-SS-0006-0-0003 D F09-SS-0006-0-0003 D F09-SS-0006-0-0003 D F09-SS-0006-0-0003 D F09-S S -0006-0-0010 D F09-SS-0006-0-0010 D F09-S S -0006-0-0010 D F09-S S -0006-0-0010 D F09-SS-0007-0-0000 D F09-SS-0007-0-0000 D F09-SS-0007-0-0000 D F09-SS-0007-0-0000 Am ount S p ik e d (n g /g ) 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 13c -p f o a Am ount R ecovered W e t W eig ht (ng /g ) NR NR NR NR NR NR NR NR 5 .1 9 5 .1 9 3 .0 0 306 5.61 5 .3 4 2 .8 5 294 5 .3 9 5 .1 4 3 .9 4 314 5 .3 8 4 .9 4 3 .0 9 291 5 .2 3 5.21 4 .0 2 390 6 .0 4 5 .6 2 3 .8 3 348 5.81 5 .5 8 3 .0 0 348 2 .8 6 2 .5 5 1.74 261 R ecovery (% ) NR NR NR NR NR NR NR NR 130 130 75 77 140 134 71 74 135 129 99 79 135 124 77 73 131 130 101 98 151 141 96 87 145 140 75 87 72 64 44 65 Note: Since this sum m ary table shows rounded results, recovery values m ay vary slightly from the values in the raw data. N R = Not Reported due to Quality Control failures, see Table V III for re-extract data. Exygen Research Page 56 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table VII. Surrogate Spike Recovery of 13C PFOA in Soil Samples Continued Exygen ID Sam ple D escription C0160882 C0160882 Rep C 0160882 Spk E C 0160882 Spk F C0160883 C0160883 Rep C 0160883 Spk G C 0160883 Spk H CO160884 CO160884 Rep C0160884 Spk I C0160884 Spk J CO160885 CO160885 Rep C0160885 Spk K C 0160885 Spk L CO160886 CO160886 Rep C 0160886 Spk C C0160886 Spk D CO160887 C0160887 Rep C0160887 Spk E C 0160887 Spk F CO160888 CO160888 Rep CO160888 Spk G C0160888 Spk H CO160889 CO160889 Rep C0160889 Spk C C0160889 Spk D C0160890 CO160890 Rep C0160890 Spk E C0160890 Spk F C 0160891 C0160891 Rep C0160891 Spk G C0160891 Spk H D F09-SS-0007-0-0003 D F09-SS-0007-0-0003 D F09-SS-0007-0-0003 D F09-SS-0007-0-0003 D F09-S S -0007-0-0010 D F09-S S -0007-0-0010 D F09-S S -0007-0-0010 D F09-S S -0007-0-0010 D F09-SS-0007-D B-0010 D F09-S S -0007-D B -0010 D F09-S S -0007-D B -0010 D F09-SS-0007-D B-0010 D F09-SS-0008-0-0000 D F09-SS-0008-0-0000 D F09-SS-0008-0-0000 D F09-SS-0008-0-0000 D F09-SS-0008-0-0003 D F09-SS-0008-0-0003 D F09-SS-0008-0-0003 D F09-SS-0008-0-0003 D F09-SS-0008-0-0010 D F09-S S -0008-0-0010 D F09-S S -0008-0-0010 D F09-S S -0008-0-0010 D F09-SS-0009-0-0000 D F09-SS-0009-0-0000 D F09-S S -0009-0-0000 D F09-SS-0009-0-0000 D F09-SS-0009-0-0003 D F09-SS-0009-0-0003 D F09-SS-0009-0-0003 D F09-SS-0009-0-0003 D F09-SS-0009-D B-0003 D F09-SS-0009-D B-0003 D F09-SS-0009-D B-0003 D F09-SS-0009-D B-0003 D F09-SS-0009-0-0010 D F09-SS-0009-0-0010 D F09-S S -0009-0-0010 D F09-S S -0009-0-0010 Am ount S p ik e d (n g /g ) 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 13c -p f o a Am ount R ecovered W e t W eig ht (ng /g ) 3 .0 7 3 .0 0 1.70 240 4 .2 6 4 .0 5 2 .2 4 251 4 .8 8 4 .9 3 3 .5 0 395 2 .6 8 2.61 2 .0 3 369 2 .1 0 2 .3 5 1.39 278 2 .7 9 3 .0 4 1.84 330 5 .1 0 5 .2 2 3 .7 0 429 5.11 5.11 3 .4 6 480 4 .9 4 5 .0 0 3 .4 2 391 5 .1 0 4 .8 2 2 .9 2 413 R ecovery (% ) 77 75 43 60 107 101 56 63 122 123 88 99 67 65 51 92 53 59 35 70 70 76 46 83 128 131 93 107 128 128 87 120 124 125 86 98 128 121 73 103 Note: Since this sum m ary table shows rounded results, recovery values m ay vary slightly from the values in th e raw data. NR = Not Reported due to Quality Control failures, see Table V III for re-extract data. Exygen Research Page 57 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table VII. Surrogate Spike Recovery of 13C PFOA in Soil Samples Continued Exygen ID Sam ple D escription CO160892 CO160892 Rep CO160892 S p k I C 0160892 Spk J CO160893 C 0160893 Rep C 0160893 Spk K C0160893 Spk L C0160894 CO160894 Rep C0160894 Spk I C 0160894 Spk J CO160895 C0160895 Rep C 0160895 Spk K C 0160895 Spk L CO160896 C0160896 Rep C 0160896 Spk C C 0160896 Spk D CO160897 CO160897 Rep CO160897 S pk E C0160897 Spk F C 0160898 CO160898 Rep C0160898 Spk G C0160898 Spk H CO160899 C0160899 Rep C0160899 Spk C C 0160899 Spk D CO160900 C0160900 Rep C 0160900 Spk E C 0160900 Spk F C 0160901 C0160901 Rep C0160901 Spk G C0160901 Spk H D F09-S S -0010-0-0000 D F09-S S -0010-0-0000 D F09-S S -0010-0-0000 D F09-S S -0010-0-0000 D F09-SS-0010-0-0003 D F09-SS-0010-0-0003 D F09-SS-0010-0-0003 D F09-SS-0010-0-0003 D F09-S S -0010-0-0010 D F09-S S -0010-0-0010 D F09-SS-001Q -0-0010 D F09-S S -0010-0-0010 D F 09-S S -0011-0-0000 D F 09-S S -0011-0-0000 D F 09-S S -0011-0-0000 D F 09-S S -0011-0-0000 D F09-S S -0011-0-0003 D F 09-S S -0011-0-0003 D F 09-S S -0011-0-0003 D F 09-S S -0011-0-0003 D F 09-S S -0011-0-0010 D F 09-S S -0011-0-0010 D F 09-S S -0011-0-0010 D F09-S S -0011-0-0010 D F09-S S -0012-0-0000 D F09-S S -0012-0-0000 D F09-SS-0012-0-0000 D F09-SS-0012-0-0000 D F09-SS-0012-D B-0000 D F09-SS-0012-D B-0000 D F09-SS-0012-D B-0000 D F09-SS-0012-D B-0000 D F09-SS-0012-0-0003 D F09-SS-0012-0-0003 D F09-SS-0012-0-0003 D F09-SS-0012-0-0003 D F09-SS-0012-0-0010 D F09-SS-0012-0-0010 D F09-SS-0012-0-0010 D F09-SS-0012-0-0010 Am ount S p ik e d (n g /g ) 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 13c - p f o a Am ount R ecovered W e t W eig ht (n g /g ) 3 .2 6 3 .2 6 2 .1 4 370 4 .0 2 3 .3 4 2 .4 0 330 3 .1 0 4 .3 0 2 .4 4 349 3 .7 3 3 .3 5 2 .1 3 298 3 .4 5 3 .2 9 1.64 275 NR NR NR NR NR NR NR NR 3 .9 5 4 .1 0 2 .4 9 365 NR NR NR NR 4 .0 2 4 .2 2 2 .1 5 326 R ecovery (% ) 82 82 54 93 101 84 60 83 78 108 61 87 93 84 53 75 86 82 41 69 NR NR NR NR NR NR NR NR 99 103 62 91 NR NR NR NR 101 106 54 82 Note: Since this sum m ary table shows rounded results, recovery values m ay vary slightly from the values in the raw data. N R = Not Reported due to Quality Control failures, see Table VIII for re-extract data. Exygen Research Page 58 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table VII. Surrogate Spike Recovery of 13C PFOA in Soil Samples Continued Exygen ID C0160902 C0160902 Rep C0160902 Spk I C0160902 Spk J CO160903 C0160903 Rep C0160903 Spk K C 0160903 Spk L CO160904 C0160904 Rep C 0160904 Spk C C0160904 Spk D CO164247 C0164247 Rep C0164247 Spk K C 0164247 Spk L CO164248 C0164248 Rep C 0164248 Spk C C 0164248 Spk D CO164249 CO164249 Rep C0164249 Spk E C0164249 Spk F C 0164250 C0164250 Rep C0164250 Spk G C0164250 Spk H C0164251 C0164251 Rep C0164251 S pkI C0164251 S pkJ CO164252 CO164252 Rep C0164252 Spk K C 0164252 Spk L C0164253 CO164253 Rep C0164253 Spk C C0164253 Spk D Sam ple D escription D F09-SS-0013-0-0000 D F09-SS-0013-0-0000 D F09-SS-0013-0-0000 D F09-SS-0013-0-0000 D F09-SS-0013-0-0003 D F09-S S -0013-0-0003 D F09-S S -0013-0-0003 D F09-S S -0013-0-0003 D F09-S S -0013-0-0010 D F09-SS-0013-0-0010 D F09-S S -0013-0-0010 D F09-SS-0013-0-0010 D AL-SB-602R -0-0000 D AL-SB-602R -0-0000 D AL-SB-602R -0-0000 D AL-SB-602R -0-0000 D AL-SB-602R -0-0005 D AL-SB-602R -0-0005 D AL-SB-602R -0-0005 D AL-SB-602R -0-0005 D AL-SB-602R -0-0010 D AL-SB-602R -0-0010 D AL-SB-602R -0-0010 D AL-SB-602R -0-0010 D AL-SB-602R -0-0050 D AL-SB-602R -0-0050 D AL-SB-602R -0-0050 D AL-SB-602R -0-0050 D AL-SB-602R -0-0100 D AL-SB-602R -0-0100 D AL-SB-602R -0-0100 D AL-SB-602R -0-0100 D AL-SB-601R -0-0000 D AL-SB-601R -0-0000 DAL-SB-601 R -0-0000 DAL-SB-601 R -0-0000 DAL-SB-601 R -D B-0000 DAL-SB-601 R -D B-0000 DAL-SB-601 R-D B-0000 DAL-SB-601 R-D B-0000 Am ount S p ik e d (n g /g ) 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 13c -p f o a Am ount R ecovered W e t W eig ht (ng /g ) 4 .5 0 4 .8 2 2 .8 9 326 5 .1 4 5 .3 2 2 .3 7 357 4 .4 3 4 .3 8 2 .2 2 245 5 .4 0 5 .1 7 2 .9 0 299 5 .3 4 5 .2 2 2 .8 6 309 5 .8 9 5 .7 8 4 .3 0 408 5 .4 0 4 .8 7 2 .6 9 295 3 .9 7 4 .6 8 2 .5 8 276 5 .3 6 5 .2 6 2 .7 2 390 7.01 5 .7 5 4 .0 2 473 R ecovery (% ) 113 121 72 82 129 133 59 89 111 110 56 61 135 129 73 75 134 131 72 77 147 145 108 102 135 122 67 74 99 117 65 69 134 132 68 98 175 144 101 118 Note: Since this sum m ary table shows rounded results, recovery values m ay vary slightly from the values in the raw data. N R = Not Reported due to Quality Control failures, see Table V III for re-extract data. Exygen Research Page 59 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table VII. Surrogate Spike Recovery of 13C PFOA in Soil Samples Continued Exygen ID C0164254 C0164254 Rep C0164254 Spk E C 0164254 Spk F CO164255 C0164255 Rep C 0164255 Spk G C0164255 Spk H C0164256 C0164256 Rep C0164256 S pkI C0164256 Spk J CO164257 C0164257 Rep C0164257 Spk K C 0164257 Spk L C0164259 C0164259 Rep C0164259 Spk C C0164259 Spk D CO164260 C0164260 Rep C0164260 Spk E C 0164260 Spk F Sam ple D escription D AL-SB-601R -0-0005 D AL-SB-601R -0-0005 DAL-SB-601 R -0-0005 DAL-SB-601 R -0-0005 DAL-SB-601 R -0-0010 DAL-SB-601 R -0-0010 DAL-S B-601R -0-0010 DAL-SB-601 R-0-0010 DAL-SB-601 R -D B-0010 DAL-SB-601 R -D B-0010 DAL-SB-601 R -D B-0010 DAL-SB-601 R -D B-0010 DAL-SB-601 R -0-0050 DAL-SB-601 R -0-0050 DAL-SB-601 R -0-0050 DAL-SB-601 R -0-0050 DAL-SB-601 R -0-0100 DAL-SB-601 R -0-0100 DAL-SB-601 R -0-0100 DAL-S B-601R -0-0100 D A L-S B -601R -D B -0100 DAL-S B -6 0 1R -D B -0100 D AL-SB-601 R -D B -0100 DAL-SB-601 R -D B -0100 Am ount S p ik e d (n g /g ) 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 4 4 4 400 I3c -p f o a Am ount R ecovered W e t W eig ht (ng /g ) 5 .8 8 5 .6 0 3 .1 6 397 NR NR NR NR 5 .8 9 5 .5 4 3 .7 0 499 5 .1 0 4 .9 3 2 .7 9 269 5 .9 6 5.91 3 .2 9 352 5 .7 8 5.41 2.81 322 R ecovery (% ) 147 140 79 99 NR NR NR NR 147 139 93 125 128 123 70 67 149 148 82 88 145 135 70 81 A ve ra g e : Standard Deviation: 91 32 Note: Since this summary table shows rounded results, recovery values may vary slightly from the values in the raw data. NR = Not Reported due to Quality Control failures, see Table V III for re-extract data. Exygen Research Page 60 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table VIII. Surrogate Spike Recovery of 13C PFOA in Re-extracted Soil Samples Exygen ID C0156344 C0156344 S pkE C0156345 C0156345 Spk G C 0 156351 C0156351 Spk M C0156352 C0156352 Spk E C0156353 C0156353 Spk G C0156354 C0156354 Spk I C0156367 CO156367 Spk N C0156368 C0156368 S pkE C0156370 C0156370 Spk G C 0 156371 C0156371 Spk I C0156373 C0156373 Spk M C 0 156381 C0156381 Spk I CO156383 C0156383 Spk M C0156384 C0156384 Spk E CO156385 C0156385 Spk G C0156386 C0156386 Spk I C0156388 C0156388 Spk M C0156389 C0156389 Spk E C0156393 C0156393 SpkM Sam ple D e scrip tio n D F 1 3 -S S -S S 0 3 -0 -0 0 0 0 D F 1 3 -S S -S S 0 3 -0 -0 0 0 0 D F 1 3 -S S -S S 0 3 -0 -0 0 0 3 D F 1 3 -S S -S S 0 3 -0 -0 0 0 3 D F 1 3 -S S -S S 0 5 -0 -0 0 0 0 D F 1 3 -S S -S S 0 5 -0 -0 0 0 0 D F 1 3 -S S -S S 0 5 -0 -0 0 0 3 D F 1 3 -S S -S S 0 5 -0 -0 0 0 3 D F 13 -S S -S S 05 -D B -00 03 D F 1 3 -S S -S S 0 5 -D B -0 0 0 3 D F 1 0 -S S -S S 0 1 -0 -0 0 0 0 DF 10-SS-SS01-0-0000 DF 11-SS-SS01-0-0003 DF 11-SS-SS01-0-0003 D F 1 1-S S -S S 0 2 -0 -0 0 0 0 D F 1 1 -S S -S S 0 2 -0 -0 0 0 0 D F 1 1-S S -S S 0 2 -0 -0 0 0 3 D F 1 1-S S -S S 0 2 -0 -0 0 0 3 D F 1 1 -S S -S S 0 3 -0 -0 0 0 0 D F 1 1-S S -S S 0 3 -0 -0 0 0 0 D F 1 1 -S S -S S 0 3 -0 -0 0 0 3 D F 1 1 -S S -S S 0 3 -0 -0 0 0 3 D F 0 5 -S S -S S 0 2 -0 -0 0 0 3 D F 0 5 -S S -S S 0 2 -0 -0 0 0 3 DF 12-SS-SS01-0-0003 D F 12 -S S -S S 01-0 -0 0 0 3 D F 1 2 -S S -S S 0 2 -0 -0 0 0 0 D F 1 2 -S S -S S 0 2 -0 -0 0 0 0 D F 1 2 -S S -S S 0 2 -0 -0 0 0 3 D F 1 2 -S S -S S 0 2 -0 -0 0 0 3 DF 12-SS-SS03-0-0000 D F 1 2 -S S -S S 0 3 -0 -0 0 0 0 DF 12-SS-SS04-0-0000 DF 12-SS-SS04-0-0000 D F 1 2 -S S -S S 0 4 -0 -0 0 0 3 D F 12 -S S -S S 04 -0-00 03 DF 13-SS-SS01-0-0003 DF 13-SS-SS01-0-0003 A m ount Spiked (n g /g ) 4 400 4 400 4 2000 4 2000 4 2000 4 400 4 400 4 2000 4 800 4 400 4 400 4 400 4 400 4 2000 4 4000 4 40 4 8000 4 20000 4 400 13c -p f o a Am ount R e cove red W et W eight (n g /g ) 0.639 113 0.268 87.2 0.468 832 0.550 2170 0.438 792 NR NR 0.928 153 NR NR 0.594 425 1.50 215 NR NR 1.80 654 1.34 214 1.04 1010 0.928 1810 4 .0 5 21.4 1.13 6380 1.12 23000 1.34 263 R e co ve ry (% ) 16 28 7 22 12 42 14 109 11 40 NR NR 23 38 NR NR 15 53 38 54 NR NR 45 164 34 54 26 51 23 45 101 54 28 80 28 115 34 66 N R = Not reported due to quality control failures Note: Since this summary table shows rounded results, recovery values may vary slightly from the values in the raw data. Exygen Research Page 61 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table VIII. Surrogate Spike Recovery of 13C PFOA in Re-extracted Soil Samples Continued Exygen ID C0156394 C0156394 Spk E C0156395 C 0156395 Spk G C0159446 C0159446 Spk E CO159449 C0159449 Spk M C0159453 C0159453 Spk I C0159454 C0159454 Spk K C0159468 C0159468 Spk G C0160871 C0160871 S p kI CO160872 C 0160872 Spk K CO160873 C 0160873 Spk M CO160897 C0160897 Spk K C0160898 C 0160898 Spk M CO160900 C 0160900 Spk G CO164255 C0164255 Spk 1 Sam ple D escription D F 13-SS-SS 02-0-0000 D F 13-S S -S S 02-0-0000 DF 13-S S -S S 02-0-0003 DF 13-S S -S S 02-0-0003 D AL-SB-603R -0-0000 D AL-SB-603R -0-0000 D AL-SB-603R -0-0050 D AL-SB-603R -0-0050 D AL-SB-604R -0-0000 D AL-SB-604R -0-0000 D AL-SB-604R -0-0005 D AL-SB-604R -0-0005 D AL-SB-138R -0-0050 D AL-SB-138R -0-0050 D F 0 5 -S S -S S 0 3 -0 -0 0 0 0 D F 0 5 -S S -S S 0 3 -0 -0 0 0 0 D F05-SS-SS03-0-0003 D F 0 5 -S S -S S 0 3 -0 -0 0 0 3 D F 0 5 -S S -S S 0 4 -0 -0 0 0 0 D F 0 5 -S S -S S 0 4 -0 -0 0 0 0 D F09-S S -0011-0-0010 D F09-S S -0011-0-0010 D F09-S S -0012-0-0000 D F09-S S -0012-0-0000 D F09-SS-0012-0-0003 D F09-SS-0012-0-0003 D A L-S B -601R -0 -0010 D AL-SB-601R -0-0010 Am ount S p ik e d (n g /g ) 4 4000 4 4000 4 4 4 4 4 4 4 4 4 400 4 400 4 400 4 4 4 400 4 400 4 400 4 4 13c -p f o a Am ount R ecovered W et W eig ht (ng /g ) 1.08 2160 0 .8 8 8 2680 NR NR 3 .4 2 1.79 3 .0 6 1.54 3 .1 8 2 .4 6 2 .4 0 218 1.33 162 NR NR NR NR 2 .1 0 154 NR NR NR NR NR NR R ecovery (% ) 27 54 22 67 NR NR 86 45 77 39 80 62 60 55 33 41 NR NR NR NR 53 39 NR NR NR NR NR NR A v e ra g e : Standard D eviation: 48 30 NR = N ot reported due to q u a lity con tro l failure s Note: Since this summary table shows rounded results, recovery values may vary slightly from the values in the raw data. Exygen Research Page 62 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table IX. Surrogate Spike Recovery of 13C PFOA in Associated Rinse Blanks and Trip Blanks Exygen ID C 0156348 C 0156349 C 0156350 C 0156359 C 0156369 C0159443 C0159444 C0159445 C 0159452 C 0159465 C 0159469 C 0160867 C0160868 C 0160869 C 0160870 C0164222 C0164223 C0164224 C0164258 S a m p le D e s c rip tio n D F 13 -S S -T R IP 1 -0-0 602 21 D F 13 -S S -T R IP 1-L S -0 6 0 2 2 1 D F 13 -S S -T R IP 1 -H S -060221 D F 10-S S -S S 03-R B -0000 D F 11-S S -S S 02-R B -0003 D A L -S B -T R IP 1-0 -06 030 2 D A L -S B -T R IP 1 -L S -0 6 0 3 0 2 D A L -S B -T R IP 1 -H S -0 6 0 3 0 2 D A L-S B -603R -R B -0100 D A L-S B -138R -R B -0000 D A L-S B -138R -R B -0100 D F 0 5 -S S -T R IP 2 -0 -0 6 0 3 0 8 D F 0 5 -S S -T R IP 2 -L S -0 6 0 3 0 8 D F 0 5 -S S -T R IP 2 -H S -0 6 0 3 0 8 D F 05-S S -S S 03-R B -0000 D A L -S B -T R IP -0 -0 6 0 3 1 6 D A L -S B -T R IP -L S -0 6 0 3 1 6 D A L -S B -T R IP -H S -0 6 0 3 1 6 D A L-S B -601R -R B -0100 Am ount S p ik e d (n g /L ) 13c - p f o a Am ount R e cove red (n fl/L ) R e cove ry <%) 500 1000 100000 541 875 87200 108 88 87 500 566 113 500 530 106 500 1000 100000 595 1010 99800 119 101 100 500 619 124 500 573 115 500 601 120 500 1000 100000 602 883 86500 120 88 87 500 555 111 500 1000 10000 666 998 9150 133 100 92 500 594 119 Average: 107 Standard Deviation: 14 Note: Since this summarytable shows rounded results, recovery values mayvary slightly from the values in the raw data. Exygen Research Page 63 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table X. Exygen ID C0156344 CO156345 C0156346 C0156347 C 0 156351 C0156352 C0156353 C0156354 C0156355 C0156356 C0156357 C0156358 CO156360 C 0 156361 C0156362 C0156363 C0156364 C0156365 C0156366 C0156367 C0156368 C0156370 C 0 156371 C0156372 C0156373 C0156374 C0156375 C0156376 C0156377 Total Percent Solids in Soil Samples S a m p le D e s c rip tio n D F13-S S -S S 03-0-0000 D F13-S S -S S 03-0-0003 D F13-S S -S S 04-0-0000 D F13-S S -S S 04-0-0003 D F13-S S -S S 05-0-0000 D F13-S S -S S 05-0-0003 D F13-S S -S S 05-D B -0003 D F10-SS-SS01-0-0000 D F10-SS-SS01-0-0003 D F10-S S -S S 02-0-0000 D F 10-S S -S S 02-D B -0000 D F10-S S -S S 02-0-0003 D F10-S S -S S 03-0-0000 D F10-S S -S S 03-0-0003 D F10-S S -S S 04-0-0000 D F10-S S -S S 04-0-0003 D F10-S S -S S 05-0-0000 D F10-S S -S S 05-0-0003 D F11-SS-SS01-0-0000 D F 11-S S -S S 01-0-0003 D F 11-S S -S S 02-0-0000 D F 11-S S -S S 02-0-0003 D F11-S S -S S 03-0-0000 D F11-S S -S S 03-D B -0000 D F 11-S S -S S 03-0-0003 D F11-S S -S S 04-0-0000 D F11-S S -S S 04-0-0003 D F11-S S -S S 05-0-0000 D F 11-S S -S S 05-0-0003 T o ta l S o lid s (% ) 6 5 .5 4 7 6 .7 3 83.41 8 7 .2 7 7 8 .1 7 8 4 .7 0 8 3 .3 3 8 4 .2 7 8 6 .7 3 8 6 .2 9 8 6 .4 8 8 6 .5 6 8 1 .7 4 8 4 .1 6 7 1 .5 7 7 5 .6 2 8 2 .1 8 8 2 .5 4 81.11 8 3 .7 7 6 7 .8 3 7 0 .8 0 7 9 .7 2 7 9 .8 2 8 5 .2 3 8 6 .6 7 8 9 .3 8 8 3 .6 5 8 5 .1 7 Exygen Research Page 64 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table X. Total Percent Solids in Soil Samples Continued Exygen Sample ID Description Total Solids (%) C0156378 DF05-SS-SS01-0-0000 81.57 C0156379 DF05-SS-SS01-0-0003 84.09 C0156380 DF05-SS-SS02-0-0000 82.35 C0156381 C0156382 C0156383 DF05-SS-SS02-0-0003 DF12-SS-SS01 -0-0000 DF12-SS-SS01-0-0003 84.38 82.65 81.26 C0156384 DF12-SS-SS02-0-0000 68.74 C0156385 CO156386 DF12-SS-SS02-0-0003 DF12-SS-SS03-0-0000 73.68 82.19 C0156387 C0156388 C0156389 DF 12-SS-SS03-0-0003 DF12-SS-SS04-0-0000 DF12-SS-SS04-0-0003 87.57 74.29 78.13 C0156390 C0156391 DF 12-SS-SS05-0-0000 DF12-SS-SS05-0-0003 78.20 81.22 CO156392 DF13-SS-SS01 -0-0000 77.44 C0156393 DF13-SS-SS01 -0-0003 80.09 C0156394 DF13-SS-SS02-0-0000 75.65 C0156395 C0159446 C0159447 DF13-SS-SS02-0-0003 DAL-SB-603R-0-0000 DAL-SB-603R-0-0005 80.31 90.63 85.84 C0159448 C0159449 DAL-SB-603R-0-0010 DAL-SB-603R-0-0050 81.62 82.85 C0159450 C0159451 C0159453 C0159454 DAL-SB-603R-0-0100 DAL-SB-603R-DB-0100 DAL-SB-604R-0-0000 DAL-SB-604R-0-0005 83.40 78.33 89.54 82.13 C0159455 C0159456 DAL-SB-604R-0-0010 DAL-SB-604R-0-0050 81.93 88.21 Exygen Research Page 65 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table X. Total Percent Solids in Soil Samples Continued Exygen Sample C0159457 C0159458 C0159459 C0159460 C0159461 C0159462 C0159463 C0159464 C0159466 CO159467 C0159468 C0159470 C0159471 C0160871 C0160872 C0160873 C0160874 C0160875 C0160876 C0160877 CO160878 C0160879 C0160880 C0160881 C0160882 CO160883 C0160884 C0160885 Exygen Research DAL-SB-604R-0-0100 DAL-SB-605R-0-0000 DAL-SB-605R-DB-0000 DAL-SB-605R-0-0005 DAL-SB-605R-0-0010 DAL-SB-605R-0-0050 DAL-SB-605R-0-0100 DAL-SB-138R-0-0000 DAL-SB-138R-0-0005 DAL-SB-138R-0-0010 DAL-SB-138R-0-0050 DAL-SB-138R-0-0100 DAL-SB-138R-DB-0100 DF05-SS-SS03-0-0000 DF05-SS-SS03-0-0003 DF05-SS-SS04-0-0000 DF05-SS-SS04-0-0003 DF05-SS-SS05-0-0000 DF05-SS-SS05-0-0003 DF05-SS-SS05-DB-0003 DF09-SS-0006-0-0000 DF09-SS-0006-0-0003 DF09-SS-0006-0-0010 DF09-SS-0007-0-0000 DF09-SS-0007-0-0003 DF09-SS-0007-0-0010 DF09-SS-0007-DB-0010 DF09-SS-0008-0-0000 77.17 81.42 81.76 84.78 82.01 79.83 73.86 86.23 87.94 86.15 81.93 78.61 80.36 80.74 83.20 84.22 87.24 86.54 86.68 86.89 89.09 87.28 86.20 86.39 89.57 89.36 89.39 86.47 Page 66 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table X. Total Percent Solids in Soil Samples Continued Exygen ID Sample Description Total Solids (%) C0160886 DF09-SS-0008-0-0003 88.27 C0160887 DF09-SS-008-0-0010 88.54 C0160888 C0160889 DF09-SS-0009-0-0000 DF09-SS-0009-0-0003 87.96 88.35 C0160890 C0160891 DF09-SS-0009-DB-0003 DF09-SS-0009-0-0010 88.75 87.79 C0160892 DF09-SS-0010-0-0000 72.09 C0160893 C0160894 DF09-SS-0010-0-0003 DF09-SS-0010-0-0010 75.60 76.57 C0160895 DF09-SS-0011-0-0000 87.53 C0160896 C0160897 DF09-SS-0011-0-0003 DF09-SS-0011-0-0010 86.92 86.19 C0160898 C0160899 C0160900 DF09-SS-0012-0-0000 DF09-SS-0012-DB-0000 DF09-SS-0012-0-0003 90.45 90.43 88.07 C0160901 C0160902 DF09-SS-0012-0-0010 DF09-SS-0013-0-0000 87.02 87.52 C0160903 C0160904 DF09-SS-0013-0-0003 DF09-SS-0013-0-0010 87.52 85.76 C0164247 C0164248 C0164249 DAL-SB-602R-0-0000 DAL-SB-602R-0-0005 DAL-SB-602R-0-0010 77.82 82.30 84.24 C0164250 C0164251 C0164252 DAL-SB-602R-0-0050 DAL-SB-602R-0-0100 DAL-SB-601R-0-0000 82.18 83.52 89.43 C0164253 C0164254 DAL-SB-601R-DB-0000 DAL-SB-601 R-0-0005 87.98 88.75 C0164255 DAL-SB-601 R-0-0010 88.87 Exygen Research Page 67 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Table X. Total Percent Solids in Soil Samples Continued Exygen ID CO164256 CO164257 CO164259 C0164260 Sample Description DAL-SB-601R-DB-0010 D A L -S B -6 0 1R -0 -0 0 5 0 DAL-SB-601 R-0-0100 DAL-SB-601 R-DB-0100 Total Solids (%) 87.99 81.36 86.85 83.56 Exygen Research Page 68 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 FIGURES Exygen Research Page 69 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Figure 1. Typical Calibration Curve for PFOA in Methanol 041200AR P70-1131 Surfaoe Soil REVI.rdb (PFOA): "Lineai* Regression C'1 / * ' weighting): y 003 x + 2.1 2t+ 0 04(r* 0.9001) Area, counts Exygen Research Page 70 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Figure 2. Extracted Standards of PFOA in Methanol, 25 ng/mL and 50 ng/mL, Respectively XCM 13S-1 - PFOA (Standard) 413.tO S i.tam a -tam p! 2 o f 49 from 041206AR.wiff Araa: 44523coaatM Haight: ? .5 i+003 cp* RT: 11.2mia 11.23 Intensity, cps A ri: 74142 counti H ljh t 4.40+003 cp* RT'. 11.2 min 11.21 Intensity, cps Exygen Research Page 71 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Figure 3. PFOA in Methanol Reagent Blank, 50 ng/L Fortified Reagent Spk A, and 500 ng/L Fortified Reagent Spk B, Respectively R ta g m t Control - PFOA (Urntnowm) 413.0/369.0 mma -tam pla 9 o f 49 from 041206AR.wiff pomk mot fommd) 10.es Intensity, cps Tim, min J Rignt Spk A - PFOA(QC)413.07360.0 imu - simple 10 of 40 from 04120BAR.rtff A rtj: 71780 count! Hoight: 4.2S+003 ops RT: 11.2 min 11.22 Intensity, cps Intensity, ops Tim, min Rjgnt Spk B PFOA (00)413.0/389.0 smu - simple 11 of 40 from 041208AR.iff A m : 502618 counts Height' 3.76+004 ops RT: 115 min 3.0*42.041.0*4- 0.0 I r .... * 1 I " I ......"" i" 12 3 4 5 07 11.20 8 9 10 11 12 13 14 15 10 17 Tima, min Exygen Research Page 72 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Figure 4. Chromatogram Representing a Soil Sample Analyzed for PFOA (Exygen ID: C0156344, Data Set: 041206AR) Intensity, cps I C91SS344 PFOA (U n imow*) 4 ft. 036% 0 in -tam pl 24 o f 49 from t4120SAR.wiff A n a : 539244count* 4.17*+004cpt RT: 11.2 mia 4.0(43.8(43.0(43.4(43.2(43.0(42.8(42.0(42.4(42.2(42.0(41.8(41.0(41.4 1-2(41.0(48000.0 0000.0 4000.0 2000.0 0.0- 1150 10 11 12 13 14 15 10 17 Tlm(, min Exygen Research Page 73 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Figure 5. Typical Calibration Curve for 13C PFOA in Methanol 041200E Soil.rdb (13C PFOA): "Lineal" Regression CM / * * weighting): y * 478 x+ 0.00111 (r * 0.0070) Area, counts Exygen Research Page 74 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Figure 6. Extracted Standards of 13C PFOA in Methanol, 25 ng/mL and 50 ng/mL, Respectively XC041306-1. 13C-PFOA (Standard) 415.0/370.0 ama ample 2 o f 3 * from 9413i6A.wiff Area: 25S2 eoaate Height: 2 2 3 t+ M 3 c p * RT: 11.2 m ia 1120 Intensity, cps XC041300-2- 13C-PFOA(Stindjrd) 415.0/370.0 imu simple 3 of 38 torn 041305A.wiff Arei: 55340 counts Height: 4.00e+003 cps RT: 11.2 min 11.20 Intensity, cps Exygen Research Page 75 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Figure 7. 13C PFOA in Methanol Reagent Blank, 50 ng/L Fortified Reagent Spk A, and 500 ng/L Fortified Reagent Spk B, Respectively Intensity, cps I Raeitt Control - 13C-PFQA (Unknown) 475LA376L amu tam p/9 9of39from 04130A.wfff p s not found) 11.20 JO.12 s a on 5.eo 7 TO 10.81 -20 8 0 10 Tima, min Raagant Spk A* 13C-PFOA(QC) 415.0/370.0 jmu*sampla 10 of 38 from 041306A.wiff Area: 60001 counts Haight 5.42a+003 cps RT; 11.2 min 11.21 13.0D0U ^ 1i-4r ..6v r7 ,,,,1I5W..2A1I 17x.62 A/A/y>A>,A^Aa ^ _/vt /^v 3 14 15 16 17 Intensity, cps Tima, min Raagant Spk B - 13C-PFOA <GC) 415.0/370.0 am u-um pla 11 of 38 from 041306A.wiff Araa: 553376 counts Haight: 4.86a+004 ops RT:11.2 min 11.23 Intensiv, cps Exygen Research Page 76 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Figure 8. Chromatogram Representing a Soil Sample Analyzed for 13C PFOA (Exygen ID: C0156363, Data Set: 041306A) Intensity, cps C4756363 - 13C-PFOA (Unknown) 415.*3 7 0 .$ nma -nnmpfo 15 o f 39 from 04130A.wiff A n n : 373541 count Hkigkt: 3.16*+004cp ftT: 11.2 min 3.0*4 2.8*4 11.22 2.0*4 2.4*4 2.2*4 2.0*4 1.8*4 1.0*41.4*41.2*4 1.0*4 8000.0 0000.0 4000.0 2000.0 0.0 8 9 10 11 12 13 14 10 10 17 Tim *, mir> Exygen Research Page 77 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 APPENDIX A Study Protocol P0000760 (Exygen Study No. P0000760) with Analytical Methods and Protocol Amendments Exygen Research Page 78 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 STUDY PROTOCOL Study Title: Analysis of Perfluorooctanoic Acid (PFOA) in Water, Soil, Sediment, Fish, Clams, Vegetation, Small Mammal Liver and Small Mammal Serum Using LC/MS/MS for the 3M Decatur Monitoring Program Exygen Protocol Number: P0000760 Performing Laboratory: Exygen Research 30S8 Research Drive State College, PA 16801 Phone: (814) 272-1039 Sponsor Representative: Michael A. Santoro Director o f Regulatory Affairs 3M B uilding 0236-01 -B -10 St. Paul, M N 55144 Phone: (651) 733-6374 Page I o f 65 Exygen Research Page 79 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 DISTRIBUTION: 1) Jaisimha Kesari, Study Director, Weston Solutions 2) John M. Flaherty, Principal Investigator, Exygen Research 3) Michael A. Santoro, Sponsor Representative, 3M Company 4) Exygen Research Quality Assurance Unit Exygen Research Page 2 o f 65 Page 80 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 PROTOCOL APPROVAL Study Title: Analysis o f Perfluorooctanoic Acid (PFOA) in Water, Soil, Sediment, Fish, Clams, Vegetation, Small Mammal Livers and Small Mammal Serum Using LC/MS/MS for the 3M Decatur Monitoring Program Exygen Protocol Number: P0000760 APPROVALS Jaisimha Kesari, Study Director Weston Solutions Michael A. Sa iro, Sponsor Representative 3M Company Exygen Research Page 5 o f 65 Page 81 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 TABLE OF CONTENTS TITLE PAGE............................................................................................................ ........................................ 1 DISTRIBUTION.................................................................................................................................................2 PROTOCOL APPROVAL..................................................................................................................................3 TABLE OF CONTENTS....................................................................................................................................4 INTRODUCTION............................................................................................................................................... 5 TEST MATERIAL.............................................................................................................................................. 5 SURROGATE FIELD SPIKE COMPOUND.................................................................................................... 5 OBJEC TIVE....................................................................................................................................................... 6 TESTING FACILITY.........................................................................................................................................6 STUDY DIRECTOR...........................................................................................................................................6 SPONSOR REPRESENTATIVE........................................................................................................................7 PRINCIPAL INVESTIGATOR..........................................................................................................................7 PROPOSED EXPERIMENTAL START AND TERMINATION DATES...................................................... 7 IDENTIFICATION AND JUSTIFICATION OF THE TEST SYSTEM.......................................................... 7 SAMPLE PROCUREMENT, RECEIPT AND RETENTION.......................................................................... 8 SAMPLE IDENTIFICATION............................................................................................................................8 ANALYTICAL PROCEDURE SUMMARY........................................................................... ........................ 9 VERIFICATION OF ANALYTICAL PROCEDURE....................................................................................... 9 METHOD FOR CONTROL OF BIAS............................................................................................................... II STATISTICAL METHODS............................................................................................................................... II GLP STATEMENT............................................................................................................................................ 11 REPORT..............................................................................................................................................................11 SAFETY AND HEALTH................................................................................................................................... 12 AMENDMENTS TO PROTOCOL.................................................................................................................... 13 DATA RECORD KEEPING.............................................................................................................................. 13 QUALITY ASSURANCE.................................................................................................................................. 14 RETENTION OF DATA AND ARCHIVING................................................................................................... 14 APPENDIX I, ANALYTICAL METHODS....................................................................................................... 15 Page 4 o f 65 Exygen Research Page 82 o f 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 INTRODUCTION The purpose o f this study is to perform analysis for perfluorooctanoic acid (PFOA) in water, soil, sediment, fish, clams, vegetation, small mammal livers and small mammal serum using LC/MS/MS for the 3M Decatur Monitoring Program. The study will be audited for compliance with EPA TSCA Good Laboratory Practice Standards 40 CFR 792 by the Quality Assurance Unit o f Exygen Research. TEST MATERIAL The test material is perfluorooctanoic acid (PFOA) and was purchased from Sigma-Aldrich. PFOA Chemical Name: Perfluorooctanoic acid Molecular Weight: 414 Lot Number: 23116HB Purity: 97.64% Transitions Monitored: 413 - 369 (for quantification) and 413 --219 (for confirmation) Structure: SURROGATE FIELD SPIKE COMPOUND The surrogate field spiking compound is 13C labeled perfluorooctanoic acid (l3C PFOA) and was purchased by 3M from Perkin-Elmer Life and Analytical Sciences. ,3C PFOA Chemical Name: 1.2-13C perfluorooctanoic acid Molecular Weight: 416 Lot Number: 3507-195 Purity: 97% Transition Monitored: 415 --370 Page 5 o f 65 Exygen Research Page 83 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 OBJECTIVE The purpose o f this study is to perform analysis for perfluorooctanoic acid (PFOA) in water, soil, sediment, fish, clams, vegetation, small mammal livers and small mammal serum for the 3M Decatur Monitoring Program using the current versions o f the following Exygen analytical methods: V0001780 V0001781 V0001782 V0001783 V0001784 V0001785 V0001786 "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS" "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS" "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Sediment by LC/MS/MS" "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS" "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Vegetation by LC/MS/MS" "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Small Mammal Liver by LC/MS/MS" "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Small Mammal Serum by LC/MS/MS" TESTING FACILITY Exygen Research 3058 Research Drive State College, PA 16801 Phone: (814) 272-1039 STUDY DIRECTOR Jaisimha Kesari P.E., DEE Weston Solutions, Inc. 1400 Weston Way West Chester, PA 19380 Phone: (610) 701-3761 Fax: (610) 701-7401 j.kesari@westonsolutions.com Page 6 of 65 Exygen Research Page 84 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 SPONSOR REPRESENTATIVE Michael A. Santoro 3M Company Director o f Regulatory Affaire 3M Building 0236-01-B-10 St. Paul, MN 55144 Phone: (651) 733-6374 PRINCIPAL INVESTIGATOR John M. Flaherty Exygen Research 3058 Research Drive State College, PA 16801 Phone: (814) 272-1039 john.flaherty@exygen.com PROPOSED EXPERIMENTAL START AND TERMINATION DATES It is proposed that the analytical portion o f this study be conducted from October 01, 2004 to December 31, 2005. The actual experimental start and termination dates will be included in the final report. IDENTIFICATION AND JUSTIFICATION OF THE TEST SYSTEM The following are the test systems for this study: Water (groundwater and surface water) Soil Sediment Fish Clams Vegetation Small Mammal Liver Small Mammal Serum The samples will be collected by Weston Solutions. The control samples will be purchased and prepared by the testing facility. Purchase and processing Page 7 o f 65 Exygen Research Page 85 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number P0000760 details for the control samples will be included in the final report associated with this study. The test systems were chosen to access the environmental impact o f PFOA in the Decatur, Alabama area. SAMPLE PROCUREMENT, RECEIPT AND RETENTION Water, soil, sediment, fish, clam, vegetation, small mammal liver and small mammal serum samples will be received at Exygen directly from Weston Solutions. The details o f sample procurement for this study are outlined in the 3M work plan entitled "Phase 2 Work Plan for Sampling Environmental Media." The number and types o f samples collected will vary depending availability in the field. The total number o f samples received and analyzed for each matrix will be documented in the final report associated with this study. Water, soil, and sediment samples will be used as received without further processing at Exygen. These samples will be stored refrigerated at 2C-8C. Fish, clam, vegetation and small mammal liver samples will be processed according to the appropriate analytical method (see Appendix I). These samples will be stored frozen at -IC C . Small mammal whole blood samples will be centrifuged in the field at the time o f collection and the serum fraction will be used for the study. Small mammal serum will be stored frozen at -10C. The receipt and processing o f the samples will be documented in the final report and raw data associated with the study. SAMPLE IDENTIFICATION Prior to analysis, each sample will be assigned a laboratory sample reference number. The reference number will be unique and will distinguish each laboratory sample that is processed throughout the analytical procedure. Chromatographic data will be identified by the laboratory sample reference number. Sample storage conditions and locations will be documented throughout the study. Page 8 o f65 Exygen Research Page 86 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 ANALYTICAL PROCEDURE SUMMARY References: V0001780: "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS" V0001781: "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS" V0001782: "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Sediment by LC/MS/MS" V0001783: "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS" V0001784: "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Vegetation by LC/MS/MS" V0001785: "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Small Mammal Liver by LC/MS/MS" V0001786: "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Small Mammal Serum by LC/MS/MS" The above methods use analytical conditions capable o f separating the isomers o f PFOA. The final report will include the isomers summed into total PFOA found. VERIFICATION OF ANALYTICAL PROCEDURE A laboratory control sample will be used for the preparation o f fortified control samples. The test substance will be made into solutions as per the method, and added to the matrices via a micropipette. For water sampling, Exygen will supply one bottle per sample collected. The bottles will be 500 mL precleaned Sci/Spec Premier wide mouth HDPE bottles. These bottles have been routinely used for PFOA sample collection at the testing facility and have been shown to be free o f PFOA. All containers used for water sample collection will be shipped to the sample location containing 100 ng o f l3C PFOA (equivalent to 500 ng/L in the final sample). Samples will be added to each container to a volumetric fill line at 200 m L. A field duplicate, a low field spike and a high field spike o f each sample will be collected. The low and high field spikes will contain PFOA as well as perfluorobutanesulfonate (PFBS), perfluorohexanesulfonate (PFHS), and perfluorooctanesulfonate (PFOS). These compounds are included in the solutions used to spike the samples. The results for PFBS, PFHS and PFOS will not be reported in this study. Exygen will supply one field blank (control water) and two field blank spikes (control water fortified with PFOA at a low Page 9 o f 65 Exygen Research Page 87 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 and high level) for every twenty samples collected. At the testing facility, each water sample (excluding field duplicates and field spikes) will be extracted in duplicate and will also be fortified at a low and high concentration with PFOA and processed through the described procedure to determine method accuracy and to check for bias. For soil, sediment, clams, and vegetation, Exygen will supply one 500 mL precleaned Sci/Spec Premier wide mouth HDPE bottle per sample collected or a zip-seal bag. All containers/bags used for sample collection will be shipped to the sample location. Samples will be added to each container or bag in the field. At the testing facility, each sample will be extracted in duplicate and will also be fortified at a known concentration with PFOA at both a low and high level and processed through the described procedure to determine method accuracy and to check for bias. 13C-PFOA will also be added to each sample in the laboratory prior to extraction at the level specified below. For small mammal liver, Exygen will supply a 50 mL polypropylene centrifuge tube. For small mammal serum, Exygen will supply a collection kit for each sample containing serum separator tubes (red top), vacutainers, needle holders and needles, transfer pipettes, and polypropylene tubes. At the testing facility, each liver and serum sample will be extracted in duplicate and will also be fortified at a known concentration with PFOA at both a low and high level and processed through the described procedure to determine method accuracy and to check for bias. 13C-PFOA will also be added to each sample in the laboratory prior to extraction at the level specified below. Low and high spiking levels for each matrix are defined below: M atrix Low PFOA High PFOA IJC -P F O A Spiking Level Spiking Level Spiking Level Water 500ng/L 5000 ng/L 500 ng/L Soil 4ng/g 40ng/g 40ng/g Sediment 4ng/g 40ng/g 40ng/g Fish lOng/g 100ng/g 100ng/g Clams lOng/g 100ng/g Vegetation 10ng/g 100ng/g lOOng/g Small Mammal Liver 10ng/g 100ng/g lOOng^g Small Mammal Serum 10ng/g 100 ng/mL 100 ng/mL Recoveries are anticipated to be between 70% and 130% o f the fortified levels; however, the exact precision and accuracy will be determined by the analysis o f the quality control samples described above. A statement of accuracy will be included in the final report. Prior to sample analysis, a laboratory control spike study will be conducted in water to justify the use o f >3C PFOA results to establish precision and accuracy o f PFOA samples. The following samples are to be prepared: Page 10 o f 65 Exygen Research Page 88 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 Two control water blanks Two control water samples fortified at 250 ng/L with >3C PFOA Two control water samples fortified at 250 ng/L with l3C PFOA and at 250 ng/L with PFOA. Two control water samples fortified at 5000 ng/L with ,3C PFOA and at 5000 ng/L with PFOA. The above samples are to be extracted and analyzed according to method V0001780 entitled "Method o f Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS." METHOD FOR CONTROL OF MAS Control o f bias will be addressed by taking representative sub-samples from a homogeneous mixture of each matrix from untreated control samples, and by analyzing at least two levels o f fortifications. STATISTICAL METHODS Statistics will be limited to those specified in the subject methods and to the calculation o f average recoveries, as applicable. GLP STATEMENT All aspects o f this study shall be performed and reported in compliance with EPA TSCA Good Laboratory Practice Standards 40 CFR 792. The final report or data package (supplied to the Sponsor) shall contain a statement that the study was conducted in compliance with current and applicable GLP standards and will outline any deviations in the study from those standards. This statement will be signed by the Study Director and Sponsor Representative. REPORT A final report will be prepared by the principal investigator or their designee at the conclusion o f the study. The report will include, but will not be limited to, the following: Page ! o f 65 Exygen Research Page 89 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 The name and address o f the Study Director, Sponsor Representative, and o f the testing facility. A statement o f GLP compliance (any related documentation, such as chain-of-custody records, must be in the study records). The signed and dated statement by the Exygen Research Quality Assurance Unit regarding dates o f study inspections and dates findings were reported to the Study Director and Management. A description o f the exact analytical conditions employed in the study. If the subject method was followed exactly, it is necessary to include only a copy o f the analytical method. Any modifications to this method will be incorporated into the report. If the method is photo-reduced, the project number and page number must be included on each page. Description o f the instrumentation used and operating conditions. All results from all sets analyzed. Control and fortified samples will be identified and the data table will include sample number and fortification level. Representative chromatograms for each analyte in each matrix, including chromatograms o f a standard and a control sample, and a chromatogram at a fortification level. The location o f the analyte peaks will be clearly identified in all chromatograms. All circumstances that may have affected the quality or integrity o f the data will be documented in the report. Locations where raw data and the final report are to be archived. Additions or corrections to the final report shall be in the form o f an amendment signed by the Study Director. The amendment shall clearly identify that part o f the report that is being altered and the reasons for the alterations. The amendment will be signed and dated by the Study Director and the Sponsor Representative. SAFETY AND HEALTH Laboratory personnel will practice good sanitation and health habits. Every reasonable precaution shall be taken to prevent inadvertent exposure of personnel and the environment to the test or reference substance(s). Page 12 o f 65 Exygen Research Page 90 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number P0000760 r * AMENDMENTS TO PROTOCOL All significant changes to the analytical protocol outlined here will be expressed in writing, signed and dated by die Study Director and Sponsor Representative. Amendments usually will be issued prior to initiation o f study plan change. However, when a change is required without sufficient time for the issue o f a written amendment, that change may be effected verbally with supporting documentation signed and dated by the Study Director and followed with a written amendment as soon as possible. In this case, the effective date o f the written amendment will be die date o f the documented change. Copies o f the signed amendments will be appended to all distributed study plan copies. The original amendment will be maintained with the original study plan. Any deviations from the study plan or from the analytical method as provided will be documented and reported promptly to the Sponsor Representative. DATA RECORD KEEPING Records to be maintained include the following (as appropriate): Sample tracking sheetfs) Sample receipt records, storage history, and chains o f custody History and preparation o f standards (stock, fortification, calibration) Description o f any modifications to the method Instrument run sheets, bench-sheets or logs Analytical data tables All chromatographic and instrumental conditions Sample extraction and analysis dates A complete listing o f study personnel, signatures and initials Chronological presentation o f all study correspondence Any other documentation necessary for the reconstruction o f the study Chromatograms-All chromatograms will contain the following: Sample identification, injection date, arrow or other indication o f the area of interest, and injection number corresponding to the run. Additionally, fortifications will include the amount o f analyte added and the sample number o f the sample that was fortified. Analytical standard chromatograms will additionally include the concentration (e.g., pg/mL). Page 13 o f 65 Exygen Research Page 91 o f 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 As part o f the documentation the following sheets will be included in each analytical set: a run sheet listing the samples to be run in the set, and an instrument conditions sheet describing the instrument type and operating conditions. All applicable requirements for reporting o f study results as per 40 CFR 792.185. QUALITY ASSURANCE The QA Unit o f Exygen Research will inspect the study at intervals adequate to assure compliance with GLP's, and will report the findings o f audits to the Study Director, Exygen Management, and the Sponsor Representative. RETENTION OF DATA AND ARCHIVING All hard copy raw data, including, but not limited to, the original chromatograms, worksheets, correspondence, and results shall be included with the data package submitted to the Study Director. These will be archived with the original study plan, amendments, final report, and all pertinent information from the Sponsor. The testing facility shall keep all electronic raw data and any instrument, equipment, and storage logs for the period o f time specified in 40 CFR 792.195. An exact copy o f the materials submitted to the study director will also be kept at Exygen Research. Exygen will obtain permission from the study director before discarding or returning samples. Exygen Research Page 14 o f 65 Page 92 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 APPENDIX I ANALYTICAL METHODS V0001780: "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS" V0001781: "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS" V0001782: "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Sediment by LC/MS/MS" V0001783: "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS" V0001784: 'M ethod o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Vegetation by LC/MS/MS" V0001785: `M ethod o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Small Mammal Liver by LC/MS/MS" V0001786: "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Small Mammal Serum by LC/MS/MS" Exygen Research Page IS o f 65 Page 93 of 151 Interim Report #22 - Analysis of Soil Samples Exygen'Study No.: P0000760 Exygen Protocol Number: P0000760 ANALYTICAL METHOD Method Number: V000178Q M ethod o f Analysis for the Determloetloo o f Perfluorooctanok A d d (PFOA) is W ater by LC/MS/MS Analytical Testing Facility: Exygen Research 3058 Research Drive State College, PA 16801 Approved By: <~ v A _________ Paul Connolly * Technical Leader, LC-MS, Exygen Research Ob*AW Dale / J o t a Flaherty / ' Vice Preeident, Operation. Exygto Rercarch Date Exygen Research Total Paget: 7 Page 16 o f65 Page 94 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 Bxyge R-- ic h Method Nunber VOOOI7BO | ............. ANALYTICAL M E T H O D ........... I Method o f Analysis for foe Determination ofPerfluorooctanoic Acid (PFOA) in Water by LC/MS/MS 1.0 Scope This method is to be employed for the isolation and quantitation o f perfluorooctanoic acid by High Performance Liquid Chromatography coupled to a tandem Mass Spectrametric Detector (LC/MS/MS) in water. 2.0 Safety 2.1 Always observe aafe laboratory practices. 2.2 Consult the appropriate MSDS before handling any chemical for proper safety precaution. 3.0 Sample Requirement 3.1 At least 40 m L o f test sample for extraction. 3.2 N o sample processing is needed for water samples. 3 J Samples stored refrigerated should b e allowed to equilibrate to room temperature. 3.4 All samples must be thoroughly mixed before being sampled for extraction. 3.5 Any samples containing particles should be centriiUged at -3 0 0 0 rpm for -5 minutea and foe supernatant used for the extraction. 3.6 Sample collection procedures will be specified in the sampling plan for this project. 4.0 Reagents and Standards 4.1 W ater-H P L C grade 4.2 M ethanol-H PLC grade 4.3 Ammonium Acetate - A.C.S. Reagent Grade 4.4 Perfluorooetanoie A d d - Sigma-Aldrich 5.0 Instrument and Equipment 5.1 A high performance liquid chromatograph capable o f pumping up to 2 advents equipped with a variable volume iqjector capable o f injecting 5-200 pL connected to a tandem Mass Spectrometer (LC/MS/MS). 5.2 A device to collect raw data for peak integration and quantitation. 5.3 Analytical balance capable o f reading to 0.00001 g. 5.4 50 mL disposable polypropylene centriftige tu b a . 5.5 15 mL disposable polypropylene centriftige tubes. 5.6 Disposable micropipets (50-100uL, 100-200uL). 5.7 125-mL LDPE narrow-mouth bottles. 5.8 2 mL clear HPLC vial kit. 5.9 Disposable pipettes. 5.10 Autopipettes (100-1000 pL and 10-100 p L ),w ith disposable tips. 5.11 Waters Sep Pak Vac 6 cc ( lg ) tC lE SPE cartridges. Page i n Page 17 of 65 Exygen Research Page 95 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 Exygea Research Method Nun^er VOOOI780 I ANALYTICAL M ETHOD Method o f Analysis for the Determination ofPerfluorooctanoic A d d (PFOA) in W ater by L C /M S/M S 5.12 SPE vacuum manifold. 5.13 CentriAige capable o f spinning 50 m L polypropylene tubes at 3000 rpm. 6.0 Chromstopaphic Syitcm 6.1 Analytical Column: Fluophaae RP (Keyatone Scientific), 2.1 mm x 50 mm. $m (P/N: 82505-052130) 6.2 Temperature: 30*C 6.3 Mobile Phase (A ): 2 raM Ammonium Acetate in Water 6.4 Mobile Phase (B ): Methanol 6.5 Gradient Program: Tim e fminl 0.0 1.0 8.0 20.0 22.5 SLA 65 65 25 25 65 SIB 35 35 75 75 35 Flow Rate fm L/m inl 0.3 0.3 0.3 0.3 0.3 6.6 Injection Volume: 15 pL (can be increased to aa m uch a 50 pL). 6.7 Quantitation: Peak Area - external standard calibration curve. 6.8 Run Tune: - 23 minutes. The above co n d itio n are intended aa a guide and m ay b e changed in order to optimize the HPLC system. 7.0 MS/MS System 7.1 Mode: Electrospray Negative M RM mode, monitoring 413 -+ 369 m/z. The above cooditiona are intended as a guide and m ay be changed in order (o optimize the MSMS system. 8.0 Preparation o f Solutions 8.1 Mobile Phase 8.1.1 2 mM ammonium acetate in water is prepared by adding 0.1S4 g o f ammonium acetate to 1000 mL o f water. Alternate volumes may be prepared. Page 3 u f" Pag* 18 o f 65 Exygen Research Page 96 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 Exygen Research Method Number V00017I0 ANALYTICAL METHOD Method o f Analysis for the Determination ofPerfluorooctanoic Acid (PFOA) in Water by L C /M S/M S 9.0 Standard Preparation 9.1 Standard Stock/Fortification Solution 9.1.1 Prepare a stock solution o f >*100 pg/taL o f PFOA b y w e i r i n g 10 mg o f analytical standard (corrected for purity) and dilute to 100 mL with methanol in a 125*mL LDPE bottle. 9.1.2 A 10 p g ta L fortification solution o f PFOA is prepared by bringing 10 mL o f the 100 pg/mL solution to a final volume o f 100 with methanol in a 125 mL LDPE bottle. 9.1.3 A 1.0 p ^ m L fortification solution o fPFO A is prepared b y bringing lo mL o f the 10 pg/raL solution to a final volume o f 100 with methanol in a 125 mL LDPE bottle. 9.1.4 A 0.1 pg/taL fortification solution o fPFO A is prepared by bringing 10 m L o f tire 1.0 pg/mL solution to a final volume o f 100 with methanol in a 125 m L LDPE bottle. 9.1.5 A 0.01 p ^ m L fortification solution o fP F O A is prepared by bringing 10 mL o f the 0.1 pg/mL solution to a final volume o f 100 with methanol in a 125 mL LDPE bottle. 9.1.6 The stock and fortification solutions are to b e stored in a refrigerator at approximately 4*C and are stable for a maximum period o f 6 months from the date o f preparation. 9.2 Standard Calibration Solutions 9.2.1 92 2 LC/MS/MS calibration standards are prepared in HPLC water. The calibration standards are processed through the extraction procedure, identical to samples. The following is a typical example: additional concentrations may be prepared as needed. Final Fortification Volume o f Concentration o f o f Fortification Volume Fortified Control Calibration Solution foob) (PL) Semole fmL) Standard foot)* Calibration Standard ID (example) 0 0 40 0 XCmmddyy-0 10 100 40 25 XCmmddyy*! 10 200 40 50 XCmmddyy-2 10 400 40 100 XCmmddyyO 100 too 40 250 XCmmddyy-4 100 200 too 400 40 40 500 1000 XCmmddyy-5 X C tn m d d w -4 9 The extracted concentration o f the calibration standard is equal to Sx its initial concentration, this to the concentratio n o f foe standard during the extraction (SPE). XC " extracted calibration standard. Pag* 4 yl'^ Page 19 o f65 Exygen Research Page 97 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 Eaygo R iiw ct Method Number VOOOI7SO [ ANALYTICAL M ETHOD Method o f Analysis for tbe Determination o f Perfluorooctanoic Acid (PFOA) in W ater by L C /M S /M S 9.2.3 9.2.4 92.6 A zero standard rotation (reagent blank) m art be prepared with each set o f standards extracted. Store all extracted calibratioa standards in 15-tnL polypropylm e tubes at2*C to6*C ,uptotw ow eeks. Alternate volumes and concentrations o f standards m ay be prepared as needed. 10.0 Batch Set Up 10.1 Each batch o f samples extracted (typically 20 o r lesa) m ust include at least one reagent control (method blank using HPLC water) and two reagent controls fortified at known concentrations (lab control spike) to verifv procedural recovery for the batch. ' 10.2 Requirements for field and laboratory duplicates and spikes will be specified in the quality assurance plan for this project 11.0 Sample Extraction 11.1 Measure 4Qm L o f sample or a portion o f sample diluted to 40 m L with water into 50 a tL polypropylene centriftige tubes (fortify as needed, replace lid and mix well). 11.2 Condition tbe C )t SPE cartridge* (1 g. 6 m L) by passing 10 m l methanol followed by 5 mL o f HPLC water (~ 2 drop/sec). D o not let column run dry 11.3 Load sample on conditioned C isSPB cartridge. Discard ehiate. 11.4 Elute with - 5 m L 100% methanol. Collect 5 t- o f ehiate into graduated 15 m L polypropylene centriftige tubes (final v o lu m e 5 mL). 11.$ Analyze samples using electrospray LC/MS/MS. 12.0 Ovomatography 12.1 Inject the same amount o f each standard, sample and fortified sample into the LC/MS/MS system. A calibration standard m ust precede and follow all analyzed samples. 12.2 Standards o f PFOA corresponding to at least five o r m ore concentration levels must be included in an analytical s e t 12.3 An entire set o f extracted calibration standards m ust be included at the beginning and at the end o f a sample s e t Extracted standards must be interspersed between every 5*10 samples. As an alternative, an entire set o f extracted calibration standards m ay be injected at the beginning o f a set followed by extracted calibration standards interspersed every 5*10 samples (to account for a ascend set o f extracted standards). In either case, extracted calibration standards must be the first and last injection in a sample set. 12.4 Use linear standard curve* for quantitation. Linear standard curves are generated for tbe analyte b y linear regression using 1/x weighting o f peak area Pit* 5 of7 Page 20 o f 65 Exygen Research Page 98 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 E x y fn Rtttarcfc Method Number VOM17*0 ANALYTICAL M ETHOD Method o f Analysis for the Determination o f Perfhuxooctanoic Acid (PFOA) in Water by L C /M S /M S v m calibration standard eoooantratioo using MassLynx 3.3 (or equivalent) software system. 12.5 Sample response should not exceed standard response*. Any samples that exceed standard responses should b e ftnther diluted and reanalyzed. 13.0 Acceptance Criteria 13.1 Chram atopam m ust show a peak o f a daughter km a t 369 amu from a parent o f 413 amu. The 413 a n t parent corresponds to the PFOA anion, while the daughter ion (369 amu) represents the loss o f carbon dioxide. 13.2 Method blanks must not contain PFOA at levels greater than the LOQ. If a blank contains PFOA at levels greater than SO ng/L, then a new blank sample must be obtained and the entire act must be re-extracted. 13.3 Recoveries o f control spikes and matrix qu k as must be between 70-130% o f their known values. I f a control spike fads outside the acceptable limits, the entire set o f samples should b e re-extracted. Any matrix spike outside 70* 130% should be evaluated by the analyst to determine if re-extraction is 13.4 Any calibration standard found to be a statistical outlier by using the Huge Error Test, may be excluded from the calculation o f the calibration curve. However, die total number o f extracted calibration standards that could be excluded must not exceed 20% o f the total num ber o f extracted standards iqjectod. 13.5 The correlation coefficient (R) tor calibration curves generated must be 20.992 (R* 20.985). I f calibration results fall outside these limits, then appropriate steps must be taken to adjust instrument operation, ami the standards o r the relevant act o f aanqrlea should be reanalyzed. 13.6 Retention timaa between standards and samples must not drift more than 4 % within an analytical run. I f retention tim e drift exceeds this limit within an analytical run then the set must be reanalyzed. 14.0 Calculations 14.1 Use the following equation to catculste the amount o f PFOA found (in ng/L, baaed on peak area) using tire standard curve (linear regression parameters) generated b y die Maas Lynx software program: PFOA found (na/LI - (Peak area - intercept) x D F slope DF factor by which die final volume was diluted, if necessary. Ptft 6 of? Page 21 of 65 Exygen Research Page 99 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 Exygn Research M sdwdNunber V00017S0 ANALYTICAL METHOD Method o f Analysis fior the Detennination o f Perflnorooctaaoic Acid (PFOA) in W ater by L C /M S/M S 14.2 For tem ples fortified with known amotmta o f PFOA prior to extraction, use the following equation to calculate the percent recovery. Recovery (% ) [total analyte found (ng/L) - analyte found in control (ng/L)] analyte added (ng/L) Exygen Research Page 7 of ? Page 22 o f 6S Page 100 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 ANALYTICALMETHOD Method Number: V0001781 M ethod o f Analysis fo r the D eterm lnatlea o f P crfluorooctanok Acid (PFOA) la Soil by LC /M S/M S Analytical Testing Facility: Exygen Research 3058 Research Drive State College, PA 16801 Approved By: r i u . C J j , _______ Paul Connolly ` Technical Leader, LC-MS, Exygen Research I) Date John Flaherty ' / Vice Prsidait, O pattione, Exygen Research . Date l Exygen Research Total Pagea: 7 Page 23 o f 65 Page 101 of 151 Merini Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 h y to w w fc MMbod Number V0001711 I ANALYTICAL M ETHOD M ethod o f Analysis for the Determ ination o f Perfiuorooctanoic A cid (PFO A ) in Soil by LC/M S/MS 1.0 Scope This method is to be employed for die isolation and quantitation o f periluorooctanoic a d d by High Performance Liquid Chromatography coupled to a tandem Mass Spectrometric Detector (LC/MS/MS) in soil. 2.0 Safety 2.1 Always observe safe laboratory practices. 2.2 Consult the appropriate MSDS before handling any chemical for proper safely precautions. 3.0 Sample Requirement 3.1 At least 13 g o f test sample for extraction. 3.2 No a m p le processing is needed for soil samples. 3.3 Sample* stored refrigerated should be allowed to equilibrate to room temperature. 3.4 All samples must be thoroughly m ixed before being sampled for extraction. 3.3 Sample collection procedures will be specified in the sampling plan for this project 4.0 Reagents and Standards 4.1 W ater-H P L C grade 4.2 Methanol - HPLC grade 4.3 Ammonium Acetate - A.C.S. Reagent Grade 4.4 Pcrfhioiooctanoic Acid - Sigma*Aldrich 3.0 Instrument and Equipment 3.1 A high performance liquid chromatograph capable o f pumping up to 2 solvents equipped with s variable volume iqjoctor capable o f injecting 3*200 pL connected to a tandem Mass Spectrometer (LC/MS/MS). 3.2 A device to collect raw data for peak integration and quantitation. 3.3 Analytical balance capable o f reading to 0.00001 g. 3.4 30 mL disposable polypropylene centrifuge tubes. 3.3 1S m L disposable polypropylene eentriAige tubes. 3.6 Disposable micropipets (SO-lOOuL, 100-200uL). 3.7 123-mL LDPE narrow*mouth bottles. 3.8 2 m L clear HPLC vial k it 3.9 Disposable pipettes. 3.10 Autopipette (100*1000 pL and 10*100 pL) with disposable tips. 3.11 Waters Sap Pak Vac 6 cc ( lg ) tC IS S P E cartridges. 3.12 SPB vacuum manifold. 5.13 Ultrasonic bath. Page 2 o f7 Page 24 o f 6$ Exygen Research Page 102 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol N um ber: P0000760 ExyfM&MMrck Method Number V0001781 I ANALYTICAL M IT H O P | Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PPOA) in Soil by LC/MS/MS 5.14 Wrist-action shaker. 5.15 Ccntriftige capable o f spinning 50 m L polypropylene tubes at 5000 rpm. 6.0 Chromatographic System 6.1 Analytical Column: Fluophase RP (Keystone Scientific), 2.1 mm x 50 mm, 5h (P/N: 82505-052130) 6.2 Temperature: 30*C 6.3 Mobile Phase (A) : 2 mM Ammonium Acetate in Water 6.4 Mobile Phase (B) : Methanol 6.5 Oradieot Program: Ti m AntiO 0.0 1.0 8.0 20.0 22.5 65 65 25 25 65 Flow Rate (m L /m inl 35 0.3 35 0.3 75 0.3 75 0.3 35 0.3 6.6 Iqjectian Volume: 15 p L (can be increased to as much as 50 pL). 6.7 Quantitation: Peak Area - external standard calibration curve. 6.8 RunTim e: - 2 3 minutes. The above conditions are intended as a guide and m ay b e changed in order to optimize die HPLC system. 7.0 MS/MS System 7.1 Mode: Electrospray Negative MRM mode monitoring 4 1 3 - 369 m/z for PFOA. The above conditions are intended as a guide and m ay be changed in order to optimize the MSMS system. 8.0 Preparation o f Solutions 8.1 Mobile Phase 8.1.1 2 mM anm onium acetate in water is prepared by adding 0.154 g o f ammonium acetate to 1000 m L o f water. Alternate volumes may be prepared. Page 3 o f 7 Page 25 o f 65 Exygen Research Page 103 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 BxygalUmrefc Metbod Number VOOOI7SI | ANALYTICAL M ETHOD | Method o f Analysis for the Determination o f Perfluorooctaaoic Acid (PFOA) in Soil by LC/MS/MS 9.0 Standard Pnparetioo 9.1 Standard Stock/Foitification Solution 9.1.1 Prepare a stock solution o f -1 0 0 jigfaiL o f PFOA by weighing 10 mg o f analytical standard (corrected fix'purity) and dilute to 100 mL with methanol in a 125-mL LDPB bottle. 9.1.2 A 10 pg/mL fortification solution o f PFOA ia prepared by bringing 10 m L o f the 100 pgfaiL solution to a final volume o f 100 with methanol in a 12S mL LDPE bottle. 9.1.3 A 1.0pg/m L fortification solution o f PFOA i* prepared by bringing 10 m L o fth e 10 ng/mL solution to a final volume o f 100 with methanol in a 125 m L LDPE bottle. 9.1.4 A 0.1 |ig/m L fortification solution o f PFOA is prepared by bringing 10 m L o f foe 1.0 pg/raL solution to a final volume o f 100 with methanol in a 125 mL LDPE bottle. 9.1.5 A 0.01 ji^ m L fortification aolution o f PFOA is prepared by bringing 10 mL o f foe 0.1 pg/mL solution to a final volume o f 100 with methanol ia a 125 mL LDPB bottle. 9.1.6 The stock and fortification solutions are to be stored in a refrigerator at approximately 4*C and are stable for a maximum period o f 6 months from foe date o f preparation. 9.2 Standard Calibration Solutions 9.2.1 922 LC/MS/MS calibration standards are prepared in HPLC water. The calibration standards are processed through the extraction procedure, identical to samples. The following is a typical example: additional concentrations may be prepared as needed. Final Concentiatico o f Fortification Solution /cobi 0 Fortification Volume of Volume Fortified Control (id.) Santole fati.} 0 40 Concentration o f Calibration Standard fact)* 0 Calibration Standard ID (example) XCramddyy-0 10 100 40 10 200 40 10 400 40 100 100 40 100 200 40 100 400 40 25 50 100 250 500 1000 XCmmddyy-l XCmmddyy-2 XCnunddyy-3 XCmmddyy"4 XCmmddyy-5 XCmmddW'6 * The extracted concentration o f foe calibration standard is equal to 8x its initial concentration, due to die concentration o f foe standard during die extraction (SPE). XC - extracted calibration standard. Pile 4 af7 Page 26 o f 65 Exygen Research Page 104 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 Enygii Pm rint Method Nuwbw V 000P8t | .. ANALYTICAL M ETH O D .............................. | Method o f A n ily d i for tbe Detennination o f Perfluorooctanoic Acid (PFOA) in Soil by L C /M S /M S 9.2.3 9.2.4 9.2.5 A zero standaid solution (reagent blank) m u it be prepared with each set o f standards extracted. Store all extracted calibration standards in 15*mL polypropylene tubes at 2*C to 6*C, up to two weeks. Alternate volumes and concentrationa o f standards m ay be prepared as 10.0 Batch Set Up 10.1 Each batch o f samples extracted (typically 20 o r less) must include at least one reagent control (method blank using 5 m L o f methanol) and two reagent controls fortified at known concentrations (lab control spike) to verify procedural recovery for the batch. 10.2 Requirements for field and laboratory duplicates and spikes will be specified in tbe quality assurance plan for this project 11.0 S an p le Extraction 11.1 Weigh 3 | o f sample into SO m L polypropylene centrifiige tubes (fortify as needed, replace lid and mix well). 11.2 Add 5 m L o f methanol and shake on a wrist action shaker f o r --15 minutes. I I J Transfer the tubes to an ultrasonic bath and sonicate for ~1$ minutes. 11.4 Bring the volume up to 40 m L with water in the SO mL polypropylene centrifiige tube. 11.5 Centrifiige f ix - 1 0 minutes a t -3 0 0 0 rpen. 11.6 Condition the C u SPE cartridges ( l g, 6 mL) by passing 10 mL methanol followed by 5 mL o f HPLC water (~ 2 drop/sec). Do not let column run dry 11.7 Load (decant) the sample on die conditioned C u SPE cartridge. Discard ehiate. 11.8 Elute with 5 m L 100% methanol. Collect S m L o f eluate into graduated 1$ m L polypropylene centrifiige tubes (final volume S mL). 11.9 Analyze samples using eloctroqiray LC/MS/MS. 12.0 Chromatography 12.1 Iq e c t the same amount o f each standard, sample and fortified sample into the LC/MS/MS system. A calibration standard must precede and follow all analyzed samples. 12.2 Standards o f PFOA con--ponding to at least five or m ore concentration levels must b e included in an analytical set. 1 2 J An entire set o f extracted calibration standards must be included at the beginning and at the end o f a sample act. Extracted standards must be interspersed between every 3-10 samples. A s in alternative, an entire set o f Page 5 of? Page 27 o f 65 Exygen Research Page 105 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 B x ftta Raaaaicfc Method Niwbw V000PII ANALYTICAL M ETHOD Method o f Analysis for the Determination o f Perfhiorooctanoic Acid (PFOA) in Soil by L C /M S/M S extracted calibration standards m ay be injected at the beginning o f a set followed by extracted calibration standards interspersed every 3-10 samples (to account for a second set o f extracted standards). In either case, extracted calibration standards must be the first and last injection in a sample set. 12.4 Use linear standard curves for quantitation. lin e a r standard curves arc generated for the analyte by linear regression using 1/x weighting o f peak area versus calibration standard concentration using M assLyux 3.3 (or equivalent) software system. 12.3 Sample responee should not exceed standard responses. Any samples that exceed standard responses should be f a th e r diluted and reanalyzed. 13.0 Acceptance Criteria 13.1 Chromatogram must show a peak o f a daughter ion a t 369 amu from a parent o f 413 amu. The 413 m i parent corresponds to the PFOA anion, while the daughter ion (369 amu) represents the loss o f carbon dioxide. 13.2 Method blanks m ust not contain PFOA at levels g re a t than foe LOQ. If a blank '"*"*" PFOA at levels p e s te r than 50 ng/L, then a new blank sample must be obtained and tiie entire set must be re-extracted. 13.3 Recoveries o f control spikes and matrix spikes m ust be between 70-130% o f their known values. I f a control spike foils outside the acceptable limits, the entire set o f samples should be re-extracted. Any matrix spike outside 70 130% should be evaluated by the analyst to determine if re-extraction is warranted. 13.4 Any calibration standard foimd to b e a statistical outlier by using the Huge Error Test, may be excluded from die calculation o f the calibration curve. However, foe total number o f extracted calibration standards that could be excluded must not exceed 20% o f the total number o f extracted standards injected. 13.5 The correlation coefficient (R) for calibration curves generated must be 20.992 (R* 20.985). I f calibration results foil outside these limits, then appropriate step m u be taken to adjust instrument operation, and the standards o r the relevant set o f samples should be reanalyzed. 13.6 Retention times between standards and samples must not drift more than 4 % within an analytical run. I f retention time drift exceeds this limit within an analytical run then the set m u be reanalyzed. Fife 6 of 7 Page 28 o f 65 Exygen Research Page 106 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 Eaygse Research Method Number V0001711 I ANALYTICAL METHOD Method o f Analysis for the Determination o f Perfluorooctaaoic Acid (PFOA) in Soil by LC/MS/MS 14.0 Calculation 14.1 Use the following equation to calculate the amount o f PFOA found (in ng/L, baaed on peak area) using foe atandard curve (linear regression parameters) generated by the Mass Lynx software program: PFOA found (ng/L) (Peak area - intercept) x DF lope DF factor by which foe final volume was diluted, i f necessary. 14.2 For samples fortified with known amounts o f PFOA prior to extraction, use foe following equation to calculate the percent recovery. Recovery ( S ) - [total analyte found (ng/L) - analyte found in control (ng/L)] ^ analyte added (ng/L) 14.3 Use the following equation to convert the amount o f PFOA found in ng/L to ng/g(ppb). PFOA found fpob) - fPFQA found fag/L) x volume extracted f0.04D1 sample weight (5 g) 14.4 Use foe following equation to calculate foe amount o f PFOA found in ppb based on dry weight PFOA found (ppb) dry w e ig h t- PFOA found (ppb) x [1 0 0 % / total solids(%)] Page 7 of 7 Page 29 o f 65 Exygen Research Page 107 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 ANALYTICAL METHOD Method Number: V0001782 M ethod of Aaalysls for the Determimitioa o f PerfiuorooeU aok A d d (PFOA) lo Sediment by LC/M5/MS Analytical Testing Facility: Exygen Research 3058 Research Drive State College, PA 16801 Approved By: V--V- c~iL Paul Connolly I Technical Leader, LC-MS, Exygen Research __ lOh-bM Due a 'J /n t U / i olothan Flaherty / f VVwice President, Operations, Exygen Research Due Exygen Research Tolel Piece: 7 Page 30 of 65 Page 108 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 Bayges Rim rrk Mrtwd Number VOOO1712 | ANALYTICAL M ETHOD Method o f Analysis for foe Determination o f Perfluorooctanoic Acid (PFOA) in Sediment by LC/MS/MS 1.0 Scope H ue mefood is to be employed for foe isolation and quantitation o f perfluorooctanoic a d d by High Performance Liquid Chromatography coupled to a tandem Mass Spectzometric Detector (LC/MS/MS) in sedim ent 2.0 Safety 2.1 Always obeerve safe laboratory practices. 2 2 Consult die appropriate MSDS before handling any chemical for proper safety precautions 3.0 Sample Requirement 3.1 At least 30 g o f test sample for extraction. 3.2 No sample processing is needed for sediment samples. 3.3 Samples stored refrigerated should be allowed to equilibrate to room temperature. 3.4 All samples must be thoroughly m ixed before being sampled for extraction. 3.5 Sample collection procedures will be specified in the sampling plan for this project. 4.0 Reagents and Standards 4.1 W ater-H P L C grade 4.2 Metiunol - HPLC grade 4.3 Acetic A d d -R eag en t grade 4.4 Ammonium A cetate-A .C .S. Reagent Grade 4.5 Perfiuorooctanoic A d d - Sigma-Aldrich 5.0 Instrument and Equipment 5.1 A high performance liquid chromatograph capable o f pumping up to 2 solvents equipped with a variable volume iqjector capable o f injecting 5-200 pL oonnected to a tandem Mass Spectrometer (LC/MS/MS). 5.2 A device to collect raw data for peak integration and quantitation. 5.3 Analytical balance capable o f reading to 0.00001 g. 5.4 50 mL disposable polypropylene ccntriftige tubes. 5.5 15 mL disposable polypropylene ccntriftige tubes. 5.6 Disposable micropipets (50-100uL, 100-200uL). 5.7 125-oL LDPE narrow-mouth bottles. 5.8 2 m L d e a r HPLC vial k it 5.9 Disposable pipettin 5.10 Autopipettee (100-1000 pL and 10-100 pL), w ith disposable tips. 5.11 Waters Sep Pak Vac 6 cc ( lg ) tC18 SPE cartridges. 5.12 SPE vacuum manifold. Pile 2 of7 Page 31 o f 63 Exygen Research Page 109 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 Einw K m u ii Method Hunter V00017*2 t ANALYTICAL M ETHOD \ Method o f Analysis for the Determination ofPerfluorooctanoic Acid (PFOA) in Sediment bv L C /M S /M S ' 3.13 Vortexer. 5.14 Wrist-action ihaker. 5.13 Centriftigs capable o f (pinning 30 m L polypropylene tubes at 3000 rpm. 6.0 Chromatographie System 6.1 Analytical Column: Fhiophase RP (Keystone Scientific), 2.1 mm x 30 mm. 3p (P/N: 82505-032130) 6 2 Temperature: 30*C 6 3 Mobile Phase (A ): 2 mM Ammonium Acetate in Water Mobile Phase (B) : Methanol Gradient Program: lincim i}) 0.0 1.0 8.0 20.0 22.5 2kA 65 65 25 25 65 Flow Rate & fmL/minl 35 0.3 35 0.3 75 0.3 75 0.3 35 0.3 6.6 Injection Volume: 15 p L (can be increased to as much as 50 jiL). 6.7 Quantitation: Peak Area ex tern al standard calibration curve. 6.8 Run Time: ~ 23 minutes. The above co nditio are intended as a guide and m ay be changed in order to optimise the HPLC system. 7.0 MS/MS System 7.1 Mods: Electrosjuay Negative M RM mode, monitoring 413 -* 369 m /z for PFOA. The above co n d itio are intended as a guide and m ay be changed in order to optimize the MSMS system. 8.0 Preparation o f Solutions 8.1 Mobile Phase 8.1.1 2 mM ammonium acetate in water is prepared by adding 0.134 g o f . ammonium aeetate to 1000 mL o f water. PageJof 7 Page 32 o f65 Exygen Research Page 110 o f 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 B innlM N ich Mnhod Number V0001712 I ANALYTICAL METHOD | Method o f Analysis for foe Determination ofPcrfluorooctanoic Acid (PFOA) in Sediment by L C /M S/M S 82 Extraction Solutions 8.2.1 1% acetic acid in water is prepared by adding 10 m L o f acetic acid to 1000 mL ofwater. Alternale volumes m ay be prepared. Standard Preparation 9.1 Standard Stock/Fortification Solution 9.1.1 Prepara a atoek solution o f - 1 0 0 pg/mL o f PFOA by weighing 10 mg o f analytical standard (corrected for purity) end dilute to 100 mL with methanol in a 125-mL LDPB bottle. 9.1.2 A 10 p ^ ra L fortification soh a k o f PFOA ia prepared b y bringing 10 m L o f the 100 p g ta L solution to a final volume o f 100 with methanol in a U S m L LDPE bottle. 9.1.3 A 1.0 pg/mL fortification solution o f PFOA is prepared by bringing 10 m L o f the 10 pg/mL solution to a final volume o f 100 with methanol in a 12S mL LDPE bottle. 9.1.4 A 0.1 pgfaiL fortification solution o f PFOA is prepared by bringing 10 m L o f the 1.0 pg/mL solution to a final volume o f 100 with methanol in 125 mL LDPE bottle. 9.1.5 A 0.01 pg/mL fortification solution o f PFOA is prepared by bringing 10 mL o f tits 0.1 pg/mL solution to a final volume o f 100 with methanol in a 125 m L LDPE bottle. 9.1.6 The stock and fortification solutions are to be stored in a refrigerator at approximately 4*C and are stable for a m aximum period o f 6 months from tiie date o f preparation. 9 2 Standard Calibration Solutions 9.2.1 LC/MS/MS calibration standards are prepared in m ethanol via dilution o fth s 0.1 pgfaiL fortification solution. 9 2 2 The following is a typical example: additional concentrations may be Concentration ofFortifieitioo Solution ine/mL) 100 100 100 to 5 2 Volume (L) 10 5 2 10 10 10 Diluted to (mL) 100 100 100 100 100 100 Fins! Concentration (nc/mLl 10.0 S.O 2.0 1.0 0.5 0.2 Paie 4 o f7 Page 33 o f 63 Exygen Research Page 111 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 B x ygw IU w erc Method Number V00017I2 | ANALYTICAL M ETHOD 1 Method o f Analysis for the Determination ofPerfluorooctanoic Acid (PFOA) in Sediment by LC/MS/MS 9.2.3 9.2.4 Store all calibration standards in 125-mL LDPE narrow-mouth bottles at 2*C to 6*C, up to six months. Alternate volumes and concentrations o f standards may be prepared as needed. 10.0 Batch Set Up 10.1 Each batch o f suoplec extracted (typically 20 o r lesa) must include at least one untreated control and two untreated controls fortified at known concentrations (lab control spike) to verify procedural recovery for the batch. 10.2 Requirements for field end laboratory duplicates and spikes will be specified in foe quality aaaurance plan for foil project 11.0 Sample Extraction 11.1 W eigh 5 g o f sample into SO m L polypropylene centrifuge tubes (fortify as needed, replace lid and mix well). 11.2 Add 35 m L o f I S acetic acid, cap, vortex and shake on a wrist action shaker for - 6 0 minutes, 1 1 J Centrifuge foe tubes at -3000 rpm for -2 0 minutes. 11.4 Condition die C u SPE cartridges (1 g. 6 m L) by pasting 10 mL methanol followed by 20 m L o f HPLC water ( - 2 drop/sec). Do not let column run dry 11.3 Load (decant) the sample on the conditioned C u SPE cartridge. Discard lusts. 11.6 Add 20 m L o f methanol to the left in the bottom o f foe 30 mL ccntrifiige tube. Cap, vortex and hake on a wrist action shaker for -3 0 minutce 11.7 Ceotriftige the tubes at -3 0 0 0 rpm for - 2 0 minutes. 11.8 Decant the methanol onto foe same SPE cartridge. Collect the eluate. 11.9 Wash foe column with 4 m L o f m ethanol Collect die eluate and add it to the eluate collected in step 11.8. 11.10 Condition a second C tsSPE cartridge (1 g, 6 m L) by passing 10 m L methanol followed b y 20 m L o f HPLC water ( - 2 drop/sec). D o not let column ran dry 11.11 Add foe methanol to -2 0 0 m L o f water and load on the second conditioned SPE cartridge. 11.12 Elute with - 3 m L 100H methanol. Collect 3 m L o f eluate into graduated 13 mL polypropylene centrifiige tubes (final volume 3 mL). 11.13 Analyze samples using electroepray LC/MS/MS. P|e3of7 Page 34 o f65 Exygen Research Page 112 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 Bqnw hww fc Method NttMfawVOOOnu ANALYTICAL METHOD Method o f Analysis fix the Determination o f Perftuorooctanoic A d d (PFOA) in Sediment by L C /M S/M S 12.0 Chromatography 12.1 Inject the u m e amount o f each standard, sample and fortified ample into the LC/MS/MS system. A calibration standard m ust precede and follow all analyzed samples. 12.2 Standards o f PFOA corresponding to at least five o r m ote concentration levels must be included in an analytical act. 12.3 An entire set o f extractad calibration standards m ust be included at the beginning and at the and o f a sample tel. Standards m ust b e interspersed between every 3-10 samples. As an alternative, an entire set o f calibration standards may be injected at the beginning o f e set followed by calibration standards interspersed every 3-10 samples (to account for a second set o f standards). In either case, calibration standards must be the first and last injection in a sample s e t 12.4 Use lin e standard curves for quantitation. Linear standard curves are generated for the analyte by linear regression using \fx weighting o f peak area versus calibration standard concentration using M assLynx 3.3 (or equivalent) software system. 12.3 Sample response should not exceed standard responses. Any samples that exceed standard responses should he Anther diluted and reanalyzed. 13.0 Acceptance Criteria 13.t Chromatogram must show a peak o f a daughter ion at 309 amu from a parent o f 413 amu. The 413 amu parent correspond to the PFOA anion, while the tM.ghw ion (369 amu) repr esent the loss o f carbon dioxide. 13.2 M ethod blanks m ust n et contain PFOA at levels greater than the LOQ. (f a blade contains PFOA at levels greater than 0.2 ng/mL, then a new blank sample must be obtained and the entire set must be re-extracted. 13.3 Recoveries o f control spikes and matrix spikes m ust be between 70-130% o f their known values. I f a control spike foUe outside the acceptable limits, the entire set o f samples should b e re-extracted. Any matrix spike outside 70 130% should be evaluated by die analyst to determine i f re-extraction is warranted. 13.4 Any calibration standard found to be a etatietical outlier by using the Huge Error Test, may be excluded from die calculation o f the calibration curve. However, the total number o f extracted calibration standards that could be exchided must not exceed 20% o f die total num ber o f tr a d e d standards iqjectad. 13.3 The conelstioo coefficient (R) for calibration curves generated must be 0.992 (R3 0.963). I f calibration resulta foil outside these limits, then appropriate steps must be taken to adjust instrument operation, end the stu d en ts or the relevant set o f samples should be reanalyzed. PageSof? Page 35 o f65 Exygen Research Page 113 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 ExytMRMMnk Method Number V0001782 I a n a ly t ic a l m e t h o d I Method o f Analysis for the Determination o f Perfhiorooctanoic Acid (PFOA) in Sediment by LC/MS/MS 13.6 Retention t h w between standards and samples must not drift more than 1 4 % within an analytical run. I f retention tim e drift exceeds this limit within an analytical n m then foe set m ust b e reanalyzed. 14.0 Calculations 14.1 Use the following equatioo to calculate the amount o f PFOA found (in ng/mL. based on peek a n a ) using die standard curve (linear regression parameters) generated by the Mass Lynx software program. PFOA found (m /m D -rP a a k ana-intercept) x DF slope DF - f e t o r by which foe final volume was diluted, i f necessary. 14.2 For samples fortified with known amounts o f PFOA prior to extraction, use the following equation to c s k uiste the percent recovery. Recovery (%) - [ total analyte found (ng/mL) - analyte found in control (ng/mL)] . ]00 analyte added (ng/mL) 14 3 Use tb s following equation to convert the amount o f PFOA found in ng/mL to ng/g (ppb). m nA A (ppM - [PFft* fam d fay/mL) * final volume (1 m U \ sample weight (S g) 14.4 Use the following equation (if necessary) to calculate foe amount o f PFOA found in ppb based on dry weight, PFOA found (ppb) dry weight - PFOA found (ppb) x [100% / total solids(%)] Pag* 7 of 7 Page 36 o f65 Exygen Research Page 114 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 ANALYTICAL METHOD Method Num ber V000I783 M ethod of Analyste for the Determ ination e f Perflnerooctaaoic Acid (PFOA) la Fish u d Clame by LC/MS/MS Analytical Testing Facility: Exygen Research 3058 Research Drive State College, PA 16801 Approved By: f - i L ____ Paul Connolly I Tedniicol Loader, LC-MS, Exygen I to m e li n Flaherty ' Vice President, Operations, Exygen Research Date *M c Dote Exygen Research Total F oga: I Page 37 o f 65 Page 115 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number P0000760 Exyfm JtMMfch Mttfaod Niunber VOOQ17I3 ANALYTICAL m e t h o d Method o f Analysis for the Determination o f PerfluoroocUnoic A cid (PFOA) in Fish and Clara* by LC/MS/MS 1.0 Scope This method is to be employed for foe isolation and quantitation o f perfluorooctanoic a d d by High Performance Liquid Chromatography coupled to a tandem M u s Spectromtrie Detector (LC/MS/MS) in fish and dam s. 2.0 Safety 2.1 Always observe safe laboratory practices. 2 2 Consult foe appropriate MSDS before handling any chemical for proper safety precautions. 3.0 Sample Requirement 3.1 At least 20 g o f test sim ple for extraction. 3.2 Samples should be processed before extraction. Place foe frozen sample in a food processor and homogenize with dry ice. Place the samples in containers and leave open in frozen storage overnight to allow for carbon dioxide mhKmtMui Seal and place the samples in frozen storage until time o f analysis. 3.3 Sample collection procedures rill be specified in the sampling plan for this project Reagents and Standards 4.1 W ater-H P L C grade 4.2 Acetonitrile - HPLC grade 4.3 Carbon (120-400 meah) - Reagent grade 4.4 M ethanol-H PLC grade 4.5 Silica g d (60-200 mesh) - Reagent grade 4.6 Florisil (60*100mesh) - Reagent grade 4.7 Superclean LC-NHj - Reagent grade 4.8 1-Octano]-HPLC grade 4.9 L*Ascorbic acid - Reagent grade 4.10 Dimethyldichloroeilane - Reagent grade 4.11 Toluene - Reagent grade 4.12 Ammonium Acetate - A.C.S. Reagent Grade 4.13 Perfluorooctanoic Acid - Sigma-Aklrich S.O Instrument and Equipment 5.1 A high performance liquid chromatograph capable o f pumping up to 2 solvents equipped with a variable volume iqjector capable o f injecting 5-200 pL connected to a tandem Mass Spectrometer (LC/MS/MS). 5.2 A device to collect raw data for peak integration and quantitation. 5.3 Analytical balance capable o f reading to 0.00001 g. Pig 2 o ft Page 38 o f 65 Exygen Research Page 116 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 E xygn RMMrefc Method NmbVOOOI7l3 I ... ANALYTICAL M ETHOD "1 Method o f Analysis for die Determination o f Perfluorooctsnoic Acid (PFOA) in Fish and Clams by LC/MS/MS 5.4 Rotary evaporator. 5.5 Tiasumizer. 5.6 125 m L pear-shaped flasks. 5.7 50 m L disposable polypropylene centriAige tube. 5.8 15 m L disposable polypropylene ceutriftge tubes. 5.9 Disposable micropipets (50-100uL, 100-200uL). 5.10 125-mL LDPE narrow-mouth bottles. 5.11 2 m L clear H P L C v ialld t 5.12 Disposable pipettes. 5.13 Autopipetlss (100-1000 pL and 10*100 pL), w ith disposable tips. 5.14 SPB tubes (20mL) (Supelco c a t no. N057177). 5.15 Wrist action shaker. 5.16 Centriftige e n a b le o f spinning 50 m L polypropylene tubes at 2000 tpm. 6.0 Chromatographic System 6.1 Analytical Column: Fluophase RP (Keystone Scientific), 2.1 mm x 50 mm, 5p (P/N: 82505-052130) 6.2 Temperature: 30*C 6.3 Mobile Phase (A) : 2 mM Ammonium Acetate in Water Mobile Phase (B ): Methanol Gradient Program: llB ftifi 0.0 1.0 8.0 20.0 22.5 2hA 65 65 25 25 65 Flow Rate fmt Jmin\ 35 0.3 35 0.3 75 0.3 75 0.3 35 0.3 6.6 byecdon Volume: 15 pL (can be increased to as much as SO pL). 6.7 Quandtedon: Peak Area - external standard calibration curve. 6.8 Run Tune: - 2 3 minutes. The above conditions are intended as a guide and may be changed in order to optimize foe HPLC system. Pas*3 of! Page 39 o f 65 Exygen Research Page 117 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 Exygaa Research Method Number V000I7IJ I ANALYTICAL M IT H O P | Method o f Analytic for the D e te n n iu tio a o f PerOuorooctenoic Acid (PFOA) in Fiih end C U nu by LC/MS/MS 7.0 MS/MS System 7.1 Mode: Bleetroepny Negative MRM mode, m onitoring 413 -* 369 m/z for PFOA. The above conditions are intended as a guide and m ay be changed in order to optimize tbe MSMS system. 8.0 Preparation o f Solutions 8.1 Mobile Phase 8.1.1 2 mM ammonium acetate in water i t prepared b y adding 0.154 g o f ammonium acetate to 1000 mL o f water. 8 J Extraction Solutions 8.2.1 8.2.2 2% ascorbic a d d in methanol is prepared by dissolving 2 g o f ascorbic acid in 100 mL o f methanol 30% Dimethyldichlorosiiane in toluene is prepared by bringing 3 mL o f dimethyldichlorosiiane to a final volume o f 10 m L with toluene. Akemate volumes may be prepared. 9.0 Standard Preparation 9.1 Standard Stock/Fortificatioa Solution 9.1.1 9.1.2 9.1.3 9.1.4 9.1.5 Prepare e stock solution o f -1 0 0 jig/mL o f PFOA by weighing 10 mg o f analytical standard (corrected for purity) and dilute to 100 mL with methanol in a 125-mL LOPE bottle. A 1.0 |ig fa L fortification solution o f PFOA is prepared by bringing 1 m L o f tiie 100 p g fa L solution to a final volume o f 100 with methanol in a 125 mL LDPE bottle. A 0.1 pg/mL fortification solution o f PFOA is prepared by bringing 10 m L o fth e 1.0 pgfrnL solution to a final volume o f 100 with methanol in a 12Sm L LDPE bottle. A 0.01 jigftnL fortification solution o f PFOA is prepared by bringing 10 mL o f tiie 0.1 pgfooL solution to a final volume o f 100 with methanol in a 125 m L LDPE bottle. The stock and fortification solutions w e to be stored in s refrigerator at approximately 4C and are stable for a maximum period o f 6 months from tiie date o f (Reparation. Faf4 ofS P a g e 40 o f 65 Exygen Research Page 118 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 E iyial<w icb Method N u o ta VOOOI783 ANALYTICAL M ETHOD Method o f Analyst* for the Detcnninatwo o f Perfluocooctaaoic A d d (PFOA) in Fish and Clams by LC/MS/MS 9 2 Standard Calibration Solutions 92.1 922 LC/MS/MS calibration standards are prepared in methanol via dilution o f the 1.0 pg/mL fortification solution. The following is a typical example: additional concentrations may be prepared as needed. Final o f Fortification Volume Diluted to Concentration Solution lus/mL) (mL) (mL) fua/m L ) 1.0 S.O 100 0.05 1.0 2.5 100 1.0 1.0 too 0.025 0.01 0.05 10 100 0.005 0.025 10 100 0.0023 0.1 0.005 10 10 100 100 0.001 0.0005 9 2 2 Store all calibration standards in 125-mL LDPE narrow-mouth bottles at 2*C to 6"C, up to six months. 9.2.4 Alternate volumes and concentrations o f standards m ay be prepared as 10.0 Batch Set Up 10.1 Bach batch o f samples extracted (typically 20 o r lees) m ust include at least one untreated control and two untreated controls fortified at known concentrations (lab control spike) to verily procedural recovery for the batch. 10.2 Requirements for field and laboratory duplicates and spikes will b e specified in foe quality assurance plan for this project. 11.0 Sample Extraction 11.1 Weigh 5 g o f frozen sample into SO m L polypropylene centrifoge tubes (fortify as needed, replace lid and mix well). 11.2 Add 30 m L o f acetonitrile and shake o n a wrist action shaker f o r- I S minutes. 11.3 Place foe tubes in s freezer for ~1 hour. 11.4 Pack and condition foe SPE tubes and silanize foe pear-shaped flasks. 11.5 Pads foe 20 m L SPE tubes in sequence with 2 g florisil, 2 g silica gel. 2 g carbon, and 1 g LC-NHj. Condition foe columns with 20 mL o f methanol, then 2 0 m L o f acetonitrile. Discard all washes. Do not allow the column to dry. 11.6 Silanize foe 125 m L pear-shaped flasks by rinsing with the 30% dimefoytdichkMoeilane in toluene solution. Rinse foe flask w ith toluene once, followed by methanol (three times). D ry foe flaeke completely before use. either by air-drying or with a stream o f nitrogen. Page 5 of K Page 4 ! o f 65 Exygen Research Page 119 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 ExygMk a o n i Method Nwofcar V0001713 I ANALYTICAL M ETHOD Method o f AnalysU for foe Determination o f Pcrfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS 11.7 Centrifoge foe SO m L polypropylene tubes containing sample at 2000 rpm for 10 minutes, 11.8 Decant foe extract on to a conditioned SPE column fitted inside foe mouth o f the pear-shaped flask. Collect foe eluate in the 125 m L ailanized pear-shape flask. 11.9 Add 10 m L o f aoetenitrile to foe eample in the $0 m L centrifoge tube. Homogenize foe frozen fot phase using e ttM um iyy for 30 seconds and rinse the tissumirer with 10 mL o f acetonitrile into the tube. 11.10 Shake foe cample again for 1 0 minutes on a wrist-action shaker. 11.11 Place foe tubes in a freezer for 1 hour more. 11.12 Centrifoge foe SO m L polypropylene tubes containing sample at 2000 rpm for10num tes. . 11.13 Decant the extract onto the same SPE column. Collect foe eluate into the same pear-shaped flask and combine with foe eluent from foe initial extraction. 11.14 Pass 20 m L o f acetonitrile through foe SPE column and combine the eluate in foe same pear-shaped flask. 11.15 Add 3-4 drops o f 1-octanol to foe extract in foe pear-shaped flask and evaporate at reduced pressure using a rotary evaporator (at < 40*C). ! 1.16 Make foe final volume, by adding 2 m L o f 2% ascorbic acid in methanol to foe peer-shaped flask and swirl to mix/dissolve. 11.17 Transfer the extracts to HPLC vials using disposable pipets. 11.18 Analyze samples using electrospray LC/MS/MS. 12.0 Chromatography 12.1 Iqject the same amount o f each standard, sample and fortified sample into the LC/MS/MS system. A calibration standard must precede and follow all analyzed samples. 12.2 Standard o f PFOA corresponding to at least five o r m ore concentration levels must be included in an analytical set. 12.3 A n entire set o f calibration standards must be included at the beginning and at the end o f a sample set. Standards must be interspersed between every 5-10 samples. As an alternative, an entire set o f calibration standards may be injected at the beginning o f a set followed by calibration standards interspersed every S-10 samples (to account for a second set o f standards). In cither case, calibration standards must b e the first and last injection in a sample se t 12.4 Use linear standard curves for quantitation. Linear standard curves are generated for foe analyte by linear regression using 1tx weighting o f peak area venue calibration standard concentration uaing MaseLynx 3.3 (or equivalent) software system. Pal 6 o f8 Page 42 o f 65 Exygen Research Page 120 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 H yinR iM R h Mtbod Number V00017S3 ANALYTICAL M ETHOD Method o f Analysis for the Determination o f Perfiuorooctanoic A cid (PFOA) in Fish and Clams by LC/MS/MS 12.5 Sample response should not exceed standard responses, Any samples that exceed standard responses should be fbrther diluted and reanalyzed. 13.0 Acceptance Criteria 13.1 Chromatogram must show peak o f a daughter ion at 369 amu from s parent o f 413 amu. The 413 am u parent corresponds to the PFOA anion, while the daughter ion (369 amu) represents the lots o f carton dioxide. 13.2 Method blanks must not contain PFOA at levels greater d u n the LOQ. If a blank contains PFOA at levels greater d u n 0.5 ppb. then a new blank sample must be obtained and the entire set must be re-extracted. 13.3 Recoveries o f control spikes and matrix spikes must be between 70-130Y* o f their known values. I f a control spike foils outside the acceptable limits, the entire set o f samples should be re-extracted. 13.4 Any calibration standard found to be a statistical outlier by using the Huge B oor Test, may be excluded from the calculation o f the calibration curve. However, the total number o f calibration standards that could be excluded must not exceed 20% o f the total num ber o f standards injected. 13.5 The cosreladoa coefficient (R) for calibration curvet generated must be 20.992 (RJ 20.985). I f calibration results foil outside these limits, then appropriate steps must be taken to adjust instrument operation, and the standards o r the relevant set o f samples should be reanalyzed. 13.6 Retention times between standards and samples must not drift more than 4 % within an analytical run. I f retention tim e drift exceeds this limit within an analytical run then the set must be reanalyzed. 14.0 Calculations 14.1 Use tiie following equation to calculate the amount o f PFOA found (in ng/mL, baaed on peak area) using the standard curve (linear regression parameters) generated by the Maas Lynx software program: PFOA found (ng/mL) - (Peak area - intercept) slope 14.2 Use the following equation to eonvert the amount o f PFOA found in ng/mL to ng/g(ppb). PFOA found fppbl - fPFOA found fng/m LI final volume (m U x DF1 sample weight (g) DF - factor by which the final volume was diluted, i f necessary. Pace 7 ora Page 43 o f 65 Exygen Research Page 121 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 Exygen Research Method Number VOOOI783 ANALYTICAL M ETHOD Method o f Analyris for die Determination o f Ferfluorooctanoic Acid (PFOA) in Fish end Clams by LC/MS/MS 14.3 For samples fortified with known amounts o f PFOA prior to extraction, use die following equation to calculate foe percent recovery. Recovery ( t t ) - [ total analyte found (ng/g) - sntlyte found in control (ng/g)] analyte added (ng/g) Exygen Research PageSori Page 44 o f 65 Page 122 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 ANALYTICAL METHOD Method Number: VQ001784 M ethod o f A a d y ib for the D eterm iuedoe of Periluoroocuootc A dd (PFOA) la Veietatiou by LC/MS/MS Analytical Testing Facility: Exygen Research 3058 Research Drive State College, PA 16801 Approved By: T i - i C - iU ___ Paul Connolly ' Technical Leader, LC-MS, Exygen Research q // ..sA / ________ Jo h n Flaherty ' Vice President, Operations, Exygen Research lo m i H D ite 4^ Date Exygen Research Total Pages: 7 Page 45 o f 65 Page 123 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 Exygso fUwarcfr Mbcd Number V0001714 a n a ly t ic a l m e t h o d Method o f Analysis for the Determination o f Pcrfluorooctanoic Acid (PFOA) in Vegetation by LC/MS/MS 1.0 Scope This method is to be employed for the isolation and quantitation o f pcrfluorooctanoic acid by High Performance Liquid Chromatography coupled to a tandem Maas Spectrometric Detector (LC/MS/MS) in vegetation. 2.0 Safety 2.1 Always observe safe laboratory practices. 2.2 Consult the appropriate MSDS before handling any chemical for proper safety precautions. 3.0 Sample Requirement 3.1 At least 20 g o f test sample for extraction. 2 2 Samples should be processed before extraction. Place the frozen sample in a food processor and homogenize with (fry ice. Place the samples in containers and leave open in frozen storage overnight to allow for carbon dioxide sublimation. Seal and place die samples in frozen storage until time o f analysis. 2 3 Sample collection procedures will be specified in the sampling plan for this project. 4.0 Reagents and Standards 4.1 W a te r-H P L C grade 4 3 Acetonitrile - HPLC grade 4.3 Carbon (120-400 mesh) - Reagent grade 4.4 M ethanol-H PLC grade 4.5 Silica gel (60-200 mesh) - Reagent grade 4.6 Floriail (60-100 mesh) - Reagent grade 4.7 Supocteaa LC-NHj - Reagent grade 4.8 1-Octanol-HPLC grade 4.9 L-Ascorbic acid - Reagent p a d s 4.10 DimethyldichlococUane - Reagent grade 4.11 Toluene - Reagent grade 4.12 Ammonium A c e tate -A .C S . Reagent Crade 4.13 Pcrfluorooctanoic Acid - Sigma-Akfrich 5.0 Tmtwmwtf fad Pjp ijim u tf 5.1 A high performance liquid chromatograph capable o f pumping up to 2 solvents equipped wife a variable volume injector capable o f injecting 5-200 pL connected to a tandem M ass Spectrometer (LC/MS/MS). 5 2 A device to collect raw data for peak integration and quantitation. 5 3 Analytical balance capable o f reading to 0.00001 g. Pag*2 of 1 Page 46 of 65 Exygen Research Page 124 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number. P0000760 B xyyalU iMwh Method Number V0001784 ANALYTICAL METHOD | Method o f Analytic for the Determination o f Periluorooctanoic Acid (PFOA) in Vegetation by LC/MS/MS 5.4 Rotary evaporator. 5.5 125 mL pear-shaped fiaaks. 5.6 50 mL diapoaable polypropylene ceatrifiige tobea. 5.7 15 m L diapoaable polypropylene ccntrifiige tubes. 5.8 Diapoaable micropipets (50-lOOuL, 100-200uL). 5.9 125-mL LDPE narrow-mouth bottles. 5.10 2 m L d e a r HPLC vial kit. 5.11 Diapocable pipettea. 5.12 Autopipettes (100*1000 p L a n d 10*100 jiLX w ith disposable tips. 5.13 SPE tubes (20mL) (Supelco c a t no. N057177). 5.14 Wriat action baker. 5.15 Centrifoge capable o f spuming 50 m L polypropylene tubes at 2000 rpm. 6.0 Chromatographic System 6. ! Analytical Column: Ffuophaae RP (Keystone Scientific), 2.1 mm x 50 mm, 5m (P/N: 82505*052130) 6 2 Temperature: 30*C 6.3 Mobile Phase (A) : 2 m M Ammonium Acetate in Water Mobile Phase (B ): Methanol Gradient Program: Time fmlnl 0.0 1.0 8.0 20.0 22.5 2LA 65 65 25 25 65 Flow Rate 5 U (mL/mlnl 35 0.3 35 0.3 75 0.3 75 0.3 35 0.3 6.6 Injection Volume; 15 pL (can b e increased to a much as 50 pL). 6.7 Quantitation: Peak A rea-ex tern al standard calibration curve. 6.8 RunTim e: ~ 23 minutes. The above conditions are intended as a guide and m ay b e changed in order to optimize the HPLC system. 7.0 MS/MS System 7.1 Mode: Blectiospray Negative M RM m ode, monitoring 413 - 369 m /z for PFOA. Page 3 of 7 Page 47 o f 65 Exygen Research Page 125 o f 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 BqgnK M N R k Method Numbor V000I7M ANALYTICAL M ETHOD Method o f Analysis for the Dstennination o f Perfluorooctanoic Acid (PFOA) in Vegetation byL C /M S/M S The above conditions ace intended u a guide and may b e changed in order to optimize tfie MSMS system. 8.0 Preparation o f Solution* 8.1.1 2 n M emmooium acetate in water is prepared by adding 0.154 g o f ammonium acetate to 1000 mL o f water. 8.2 Extraction Solutions 8.2.1 8.2.2 2% ascorbic a d d in methanol is prepared by dissolving 2 g o f ascorbic a d d in 100 mL o f methanoL 30H DimethyldichlorosUane i s toluene is prepared by bringing 3 mL o f duncthyldichlorofilane to e final volume o f 10 m L with toluene. Alternate volumes may be prepared. 9.0 Standard Preparation 9.1 Standard Stock/Fortification Solution 9.1.1 Prepare a stock solution o f 100 jrgAnL o f PFOA by weighing 10 mg o f analytical standard (corrected for purity) and dilute to 100 mL with methanol in a 125-mL LDPB bottle. 9.1.2 A 1.0 pg/mL fortification solution o f PFOA is prepared by bringing I mL o f the 100 p ^ m L solution to a final volume o f 100 with methanol in a 125 mL LDPE bottle. 9.1.3 A 0.1 pg/mL fortification solution o f PFOA i t prepared by bringing 10 m L o f the 1.0 pg/mL solution to a final volume o f 100 with methanol in a 125 m L LDPE bottle. 9.1.4 A 0.01 pfftnL fortification solution o f PFOA is prepared by bringing 10 m L o f tits 0.1 pgfaiL solution to a final volume o f 100 with methanol in a 125 m L LDPE bottle. . 9.1.5 The stock and fortification solutions are to b e stored in a refrigerator at approximately 4*C end ere stable tot a maximum period o f 6 months from die date o f preparation. 9.2 Standard Calibration Solutions 9.2.1 LC/MS/MS calibration standards are prepared in methanol via dilution o f foe 1.0 pgftnL fortification solution. rag 4 o r1 Page 48 o f 65 Exygen Research Page 126 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 I x y y a Rcwn h______________________ Mfthod Number V00017S4 I ......... ANALYTICAL M ETHOD I Method o f A n a ly st fbr the Determination o f Perfhwrooctinoic A d d (PFOA) in Vegetation byL C /M S/M S 9.2.2 The following ii a typical example: additional concentrations may be Coocentrstion o f Fortification Volume Solution fuxteiL) (mL) Diluted to (mL) Finsi Concentration (ui/mL) 1.0 5.0 100 1.0 2.5 100 0.05 0.02S 1.0 0.0S 0.025 0.1 0.003 1.0 10 10 10 10 100 100 100 100 100 0.01 0.003 0.0025 0.001 0.0005 9.2.3 Store all calibration standards in 125-mL LDPE narrow-mouth bottles at 2*C to 6*C, up to six months. 9.2.4 Alternate volumes and concentrations o f standards m ay be prepared as needed. 10.0 Batch Set Up 10.1 Bach batch o f aamples extracted (typically 20 o r less) must include at least one untreatad control and two untrtated controls fortified at known concentrations (lab control spike) to verily procedural recovery for the batch. 10.2 Requirements for field and laboratory duplicates and spikes will be specified in the quality assurance plan for this project. 11.0 Sample Extraction 11.1 W eigh S g o f frozen sample into 50 m L polypropylene centriftige tubes (fortify as needed, replace lid and mix well). 11.2 Add 30 m L o f acetonitrile and shake o n a wrist action shaker for ~ 15 minutes. 11.3 Centrifuge the 50 mL polypropylene tubes containing sample at -2 0 0 0 rpm for-lO m inutes 11.4 Pack and condition the SPE tubes and stis u z e the pear-shaped flasks. 11.5 Pack the 20 m L SPE tubes in sequence with 2 g florisit, 2 g silica gel, 2 g carbon, and 1 g LC-NHj. Condition the columns with 20 m L o f methanol, then 20 m L o f acetonitrile. Discard all washes. D o not allow the column to dry. 11.6 Silanise foe 125 m L pear-shaped flasks by rinsing with the 30% diniethykUcltiorocilane in toluene solution. Rinse the flask with toluene once, followed by methanol (three times). D ry the flasks completely before use, either by air-drying o r with a streem o f nitrogen. 11.7 Decant the extract on to a conditioned SPE column fitted inside the mouth o f the pear-shaped flask. Collect the eluate in the 125 m L silanized pear-shape Page) of? Page 49 o f 65 Exygen Research Page 127 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 BxypoRMMKli Method Number V000I7|4 ANALYTICAL m e t h o d Method o f Analysis for the Determination o f Perfluorooctanoic A cid (PFOA) in Vegetation by LC/MS/MS 11.8 Add 20 m L o f acetooitrile to the sample in the 50 m L ccntriflige tube. 11.9 foe a m p le again for - 1 0 t" 1'" 1*-- on a wrist-action shaker. 11.10 Ccntriftge foe 50 m L polypropylene tubes containing sample at -2 0 0 0 rpm fo r- 5 minutes 11.11 Decant foe extract onto the wane SPE column. Collect foe eluate into the rn n o pear-shaped flask and combine with foe eluent from the initial extraction. 11.12 Repeat stepe 11.8 through 11.11 again. 11.13 Add 3*4 drops o f 1-octanol to the extract in the pear-shaped flask and evaporate at reduced pressure using a rotary evaporator (at < 40C). 11.14 Make die final volume by adding 2 m L o f 2% ascorbic acid in methanol to the pear-ehaped flask and swirl to mix/dissolve. 11.15 Transfer the extracts to HPLC vials using disposable pipeta. 11.16 Analyze samples using electrospray LC/MS/MS. 12.0 Chromatography 12.1 Iigact foe same amount o f each standard, sample and fortified sample into the LC/MS/MS Systran. A calibration standard must precede and follow all analyzed samples. \1 2 Standards o f PFOA corresponding to at least five or m ore concentration levels must be included in an analytical s e t 12.3 An entire eel o f extracted calibration standards must be included at the hagmmng and at the end o f a sample s e t Extracted standards must be interspersed between v m y 5-10 samples. Aa an alternative, an entire act of extracted calibration standards m ay be injected t t foe beginning o f a set followed by extracted calibration standards in ten p en ed every 5-10 samples (to account for a second set o f extracted standards). In either case, extracted calibration atandrada must be the first and last injection in a sample set. 12.4 U se If a f f standard curves for quantitation. Linear standard curvet are generated for foe analyte by linear regression using 1/x weighting o f peak area versus calibration standard concentration using M aasLynx 3.3 (or equivalent) software system. 12.5 Sample response should not exceed standard responses. Any samples that exceed standard responses should b e Anther diluted and reanalyzed. 13.0 Acceptance Criteria 13.1 Chromatogram must show a peak o f a daughter ion at 369 amu from a parent o f 413 The 413 amu parent corresponds to the PFOA anion, while the UngKfw foo (369 amu) represents the loss o f carbon dioxide. PS>6of7 Page SOo f 65 Exygen Research Page 128 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 ExyfwRMMich Mrthod Number V000I7S4 ANALYTICAL METHOD Method o f Analyse for (he Determination oiPerfluorooctanoic A d d (PFOA) in Vegetation by LC/MS/MS 13.2 Method blanki must not cootain PFOA s levels greater than the LOQ. If a blank contains PFOA at levels greater than O.S ppb, then a new blank sample must be obtained and the entire set must be re-extracted. 13.3 Recoveries o f control spikes and matrix spikes must be between 70-130% o f their known values. I f a control spike falls outside the acceptable limits, the entire eat o f samples should be re-extracted. 13.4 Any calibration standard found to be a statistical outlier by using the Huge Error Test, may be excluded ftom the calculation o f foe calibration curve. However, foe total number o f calibration standards that could be excluded must not exceed 20% o f foe total number o f standards injected. 13.3 The correlation coefficient (10 for calibration curvet generated must be 20.992 (R? 20.985). I f calibration resulta foil outside these limits, then sppropriate steps must be taken to adjust instrument operatimi, and the standards or the relevant set o f samples should be reanalyzed. 13.6 Retention times between standards and samples must not drift more than 4 % within an analytical run. I f retention tim e drift exceeds this limit within an analytical nm then the set must be reanalyzed. 14.0 Calculations 14.1 Use foe following equation to calculate the amount o f PFOA found (in ng/mL, baaed on peak area) using the standard curve (linear regression parameters) generated by the Mass Lynx software program: PFOA found (ngtaL) - (P w k ltM -iB a M W ) slope 14.2 Use die following equation to convert the amount o f PFOA found in ng/mL to ug/(<ppb). p u n * found fanM - fPPOA fe.mil fn .ftn l ,W fln.1 v nl.im . M . W HF1 sample weight (g) DF foctor by which the final volume was diluted, i f necessary. 14J For samples fortified w ith known amounts o f PFOA prior to extraction, use the following equation to calculate the percent recovery. Reeovery(H )- [ total analyte found (ng/g) analyte found in control (ng/g)] analyte added (ng/g) Pajff 7 of 7 Page 5 / o f 65 Exygen Research Page 129 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 ANALYTICAL METHOD Method N um ber V0001785 M ethod o f Analysis for the Determ/eartoo o f Perfloorooctaooic Acid (PFOA) la Small Mamma! U v w by LC/MS/MS Analytical Testing Facility: Bxygeo Research 3058 Research Drive State College, PA 16801 Approved By: __ C I-- _______ Paul Connolly I Technical Leader, LC-MS, Exygen Research ____I * 4 f < K Date Date Exygen Research Total Pages: 7 Page 52 o f 65 Page 130 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol N um ber: P0000760 Exyfta Research Method Number V0001715 I ANALYHCAL METHOD Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Small Mammal Liver by LC/MS/MS 1.0 Scope This method ii to be employed for the isolation and quantitation o f perfluorooctanoic a d d by High Performance Liquid Chromatography coupled to a tandem Mass Spoetromctric Detector (LC/MS/MS) in small mammal liver. 2.0 Safety 2.1 Always observe safe laboratory practices. 2.2 Consult the appropriate MSDS before handling any chemical for proper safety precautions. 3.0 Sample Requirement 3.1 At least S g o f test sample for extraction. 3.2 Samples should be proceeded before extraction. Place the frozen sample in a food proceseor and homogenize with dry ice. Place the samples in containers and leave open in frozen storage overnight to allow for carbon dioxide sublimation. Seal and place the samples in frozen storage until time o f analysis. Alternately, i f there is an insufficient amount o f sample H e s s than 5 g), then no processing is necessary and the sample can be used as supplied. 3.3 Sample collection procedures will be specified in the sampling plan for this project 4.0 Reagents and Standards 4.1 W ater-H P L C grade 4 2 M ethanol-H PLC grade 4 2 Acetonitrile-HPLC grade 4.4 Ammonium A cetate-A .C .S . Reagent Grade 4.3 Perfluorooctanoic A d d - Sigma-Aldrich 3.1 A high perform ance liquid chromatograph capable o f pumping up to 2 solvents equipped with a variable volume iqjoctor capable o f injecting 5-20U pL connected to a tandem Mass Spectrometer (LC/MS/MS). 3.2 A device to collect raw data for peek integration and quantitation. 3.3 Analytical balance capable o f reading to 0.00001 g. 5.4 50 mL disposable polypropylene centrifiige tubes. 5.5 15 mL disposable polypropylene centrifiige tubes. 3.6 Disposable micropipets (50-100uL, 100-200uL). 3.7 125-mL LDPE narrow-mouth bottles. 5.1 2 mL clear HPLC vial kit. Page 2 of t Page 53 o f 65 Exygen Research Page 131 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 ExypaRMMid) M ethodN unb* VOOOI7SS I ........ ANALYTICAL M ETH O D Method o f Analysis for foe Determination o f Perfluorooctanoic Acid (PFOA) in Small Mammal Liver by LC/MS/MS 5.9 Disposable pipettaa. 5.10 Awtopfeettes (100*1000 rL and 10-100 pL), w ith disposable Ups. 5.11 Waters Sep Pak V sc 6 cc (Ig ) tC18 SPE cartridges. 5.12 SPE vacuum manifold. 5.15 Tissuctnizer 5.14 Wrist-action shaker. 5.15 Centriftige capable o f g in n in g 15 mL polypropylene tubes at 3000 ipm. 6.0 Chromatographic System 6.1 Analytical Column: Fluophase RP (Keystone Scientific), 2.1 mm x 50 mm, 5p (P/N: 82505452130) 62 Temperature: 30*C 6.3 Mobile Phase (A ): 2 mM Ammonium Acetate in W ater Mobile Phase (B ): Methanol Gradient Program: Tim gfm m l 0.0 1.0 8.0 20.0 22.5 2hA 65 65 25 25 65 Flow Rate & (m L/m inl 35 0.3 35 0.3 75 0.3 75 0.3 35 0.3 6.6 Injection Volume: 15 p L (can be increased to as much as 50 pL). 6.7 Quantitation: Peak Area - external standard calibration curve. 6.8 RunTim e: - 2 3 minutes. The above conditions a n intended as a guide and m ay be changed in order to optimize foe HPLC Systran. 7.0 MS/MS System 7.1 Mode: Electrospray Negative MRM m ode, monitoring 413 -* 369 m/z for PFOA. The above oonditiona a n mtraided as a guide and m ay be changed in order to optimize foe MSMS Systran. Pa|3 of7 Page 54 o f 65 Exygen Research Page 132 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 BxytmRmarefe Method Number VOO01715 I ANALYTICAL METHOD Method o f Analysis fer fee Determination o f Perfluorooctanoic Acid (PFOA) in Small Mammal Liver by LC/MS/MS 8.0 Preparation o f Solutions 8.1 Mobile Phase 8.1.1 2 m M -- antmium acetate in water is prepared by adding 0.154 g of ammonium acetate to 1000 m L o f water. Alternate volumes may be prepared. 9.0 Standard Prepar ation 9. ! Standard Stock/Fortification Solution 9.1.1 Prepare a stock solution o f *>'100 pg/m L o f PFOA by weighing 10 mg o f analytical standard (corrected for purity) and dilute to 100 mL with methanol in a 125-mL LDPE bottle. 9.12 A 1.0 pg/mL fortification solution o f PFOA is prepared by bringing I m L o f fee 100 pg/mL solution to a final volume o f 100 with methanol in a 125 mL LDPE bottle. 9.13 A 0.1 pg/mL fortification solution o f PFOA is prepared by bringing 10 mL o f fee 1.0 pg/mL solution to a final volume o f 100 with methanol in a 125 m L LDPE bottle. 9.1.4 The stock and fortification solutions are to b e stored in a refrigerator at approximately 4C and are stable for a maximum period o f 6 months from fee date o f preparation. 9.2 Standard Calibration Solutions 9.2.1 LC/MS/MS calibration standards are prepared in methanol via dilution o f the 0.1 pg/mL fortification solution. 9 X 2 The following is a typical example: additional concentrations may be Concentration Final of Fortification Volume Diluted to Concentration Solution (ni/mL) (mL) (mL) (na/mU 100 5.0 100 5.0 100 2.0 100 2.0 too 1.0 100 1.0 S.0 10 100 0.5 2.0 10 100 0.2 1.0 10 100 0.1 9.2.3 Store all calibration standards in 125-mL LDPE nanrow-mouth bottles at 2*C to 6*C, up to six months. 9.2.4 Alternate volumes and concentrations o f standards m ay be prepared as Peg*4 o f7 Page 55 of 65 Exygen Research Page 133 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 Exygen JLaaarch MMhodNanber V00QI7IJ | ANALYTICAL METHOD .... Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PPOA) in Small Mammal Liver by LC/MS/MS 10.0 Batch Set Up 10.1 Each batch o f lam plee extracted (typically 20 o r less) must include at least one untreated control and two untreated controls fortified at known concentrations Gab control spike) to verify procedural recovery for the batch. 10J Requirements for field and laboratory duplicates and spikes will be specified in the quality assm n ee plan for this project 11.0 Sample Extraction 11.1 Weigh 1 g o f sample into a SO m L polypropylene ccntrifiiga tubes (fortify as needed, replace lid and mix well). Note that alternate weights o f liver may be measured depending on the sample size available for use. 11.2 A dd w ater to the sam ple for a final volum e o f lO m L . 11.3 Homogenize sample using a risauemiiar fo r- 1 minute. 11.4 Transfer 1 m L o f the sample using a disposable pipette into a IS mL disposable centrifoge tube. 11.5 Add $ m L o f acetonitrile and shake for - 2 0 minutes on a wrist-action shaker. 11.6 Centrifoge foe tubes at -3 0 0 0 ipm for - 5 minutes. 11.7 Decant foe supernatant into a SO m L disposable centrifoge tube and add 35 m L o f water. 11.8 Condition foe Cis SPE cartridges ( l g, 6 mL) by passing 10 mL methanol followed by S m L o fH P L C water ( - 2 drop/sec). D o not let column run dry 11.9 Load foe sample on conditioned C u SPE cartridge. Discard eluate. 11.10 Elate with *2 m L o f methanol. Collect 2 m L o f eluate into a graduated 15 m L polypropylene centrifuge tube (final volume * 2 mL). 11.11 Analyze samples using electrospray LC/MS/MS. 12.0 Chromatography 12.1 Iqject foe same amount o f each standard, sample and fortified sample into the LC/MS/MS system. A calibration standard must precede and follow all analyzed samples. 122 Standards o f PFQA corresponding to at least five or more concentration levels must be included in an analytical s e t 1 2 J An entire set o f calibration standards must b s included at foe beginning and at foe end o f a sample s e t Standards must be interspersed between every 5-10 samples. As an alternative, an entire set o f calibration standards may be iryacted at the beginning o f a set followed by calibration standards interspersed every 5-10 samples (to account for a second set o f standards), in either case, calibration standards must be the first and last injection in a sample set. 12.4 Use linear standard curves for quantitation. Linear standard curves are generated for foe analyte by linear regression using 1/x weighting o f peak area Pam 5 ef7 Page 56 of 65 Exygen Research Page 134 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number P0000760 BxygnftMtarcfa M ated Nanbsr V000I7W I ANALYTICAL M ETHOD Method o f Analysis ib r die Detenmnation ofPerfluorooctanoic Acid (PFOA) in Smalt Mammal Liver by LC/MS/MS venue catibeation standard concentration using M assLynx 3.3 (or equivalent) software system. 12.3 Sample response should not exceed standard responses. Any samples that exceed standard responses should be ftuther diluted and reanalyzed. 13.0 Acceptance Criteria 13.1 13.2 13.4 13.5 13.6 Chromatogram m art show peak o f a daughter ion at 369 amu from a parent o f 413 amu. Tha 413 amu parent corr esponds to die PFOA anion, while the daughter ion (369 amu) represen ts the loss o f carbon dioxide. Method blanks must not contain PFOA at levels greater than the LOQ. If a blank contains PFOA at levels greater than 10 ng/g, then a new blank sample must be obtained and the entire set must be re-extracted. Recoveries o f control spikes and matrix spikes must be between 70-130% of their known values. I f a control spike tells outside die acceptable limits, the entire set o f samples should be re-extracted. Any matrix spike outside 70 130% should be evaluated by die analyst to determine i f re-extraction is warr anted. Any calibration standard found to be a statiaticel outlier by using the Huge Error Test, m ay be excluded from die calculation o f the calibration curve. However, the total o f calibration standards that could be excluded must not exceed 20% o f the total number o f standards injected. Tbs correlation coefficient (R) for calibration curves generated must be 20.992 (R2 20.985). I f calibration results tell outside these limits, then appropriate steps must b e taken to adjust instrument operation, and the standards o r the relevant set o f samples should be reanalyzed. Retention tunes between standards and samples m ust not drift more than 4 % within an analytical run. I f retention tim e drift exceeds this limit within an analytical run then the set must be reanalyzed. 14.0 Calculations 14.1 Use the following equation to calculate the amount o f PFOA found (in ng/mL. based on peak area) using the standard curve (linear regression parameters) generated by the Mass Lynx software program: PFOA found (ngfaiL) (Peak area - mtarcaptl x DF x aliquot factor slope DF - fa c to r by which the final volume w as diluted, if necessary. Aliquot f o e t o r - 10 Page 6 of 7 Page 57 o f 65 Exygen Research Page 135 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 ExytnK M M K k M ethodNunber V00017IJ 1 .... .......... ANALYTICAL M E T H O D ........... Method o f Analysis for the Determination o f Periluorooctanoic A cid (PFOA) in Small Manunal Liver by LC/MS/MS 14.2 For samples fortified with known amount o f PFOA prior to extraction, use the following equation to calculate the percent recovery. Recovery (H ) - [ total analyte found (ng/mL) - analyte found in control (ng/mL)] ^ analyte added (ng/mL) " 14.3 Use the following equation to convert the amount o f PFOA found in ng/mL to ng/g(ppb). PFOA found fanbl - fPFOA found faa/mLl x final volume fmLll sample weight (g) Exygen Research Fas* 7 of 7 Page 58 o f 65 Page 136 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number P0000760 ANALYTICAL METHOD Method N um ber V00017M M ethod o f Analysis for the Determ ination o f Perflnorooctaaoic A d d (PFOA) io Small Mammal Seram by LO M S/M S Analytical Testing Facility: Exygen Research 3058 Research Drive State College. PA 16801 Approved By: * \ ~ c -- tL* Paul Connolly I Technical Leader, LC-MS, Exygen Research s i//n d j / _____________ Jo h n Flaherty / Vice President, Operations, Exygen Research __ Date Due Exygen Research Tolel Pife: 7 Page 59 o f65 Page 137 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 BxyfMkMMick MMbodNuobtr V00017I6 I ANALYTICAL M ETHOD Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PPOA) in Small Mammal S o w n by LC/MS/MS 1.0 Scope Thia method if to be employed for the iaoUtioo and quantitation o f perfluorooctanoic acid by High Performance Liquid Chromatography coupled to a tandem Mass Spcctromctric Detector (LC/MS/MS) in small mammal serum. 2.0 Saftty 2.1 Always observe safe laboratory practice*. 2.2 Consult the appropriate MSDS before handling any chemical for proper safety precautions 3.0 Sample Requirement 3.1 At least 1 mL o f teat sample for extraction. 1 2 N o sample processing is needed for aerum samples. However, frozen serum samples must to allowed to completely thaw to room temperature before use. 3J Sample collection procedures will be specified in the sampling plan for this project 4.0 Reagents and Standards 4.1 W ater-H P L C grade 4 2 Methanol - HPLC grade 42 Acetonitrile-HPLC giade 4.4 Ammonium Acetate - A.C.S. Reagent Grade 4.5 Perfluorooctanoic Acid - Sigmt-Aldrich 5.0 Instrument and Equipment 5.1 A high performance liquid chromatograph capable o f pumping up to 2 solvents equipped with a variable volume ipjector capable o f injecting 5-200 pL connected to a tandem Mass Spectrometer (LC/MS/MS). 52 A device to collect raw data for peak integration and quantitation. 52 Analytical balance capable o f reading to 0.00001 g. 5.4 50 mL disposable polypropylene centrifuge tubes. 5.5 15 m L disposable polypropylene centrifuge tubes. 5.6 Disposable micropipets (SO-lOOuL, 100-200uL). 5.7 125-mL LD P6 narrow-raouth bottles. 5.8 2 mL d e a r HPLC vial kit. 5.9 Disposable pipette*. 5.10 Autopipettce (100-1000 pL and 10*100 pL), w ith disposable tips. 5.11 Waters Sep Pak Vac 6 oc (lg ) (CIS SPE cartridges. 5.12 SPE vacuum manifold. 5.13 Vortexcr. Page 2 of 7 Page 60 of 6S Exygen Research Page 138 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 Exygta Riinrcb Mttbod Number V000176 | ..................... ANALYTICAL M ETH O D ................. Method o f Analysis for the Determination ofPerfluorooctanoic Acid (PFOA) in Small Mammal Seram by LC/MS/MS 5.14 Wrist-actioa shaker. 5.15 CeotriAige capable o f spinning 15 m L polypropylene tubes at 3000 rpm. 6.0 Chromatographic System 6.1 AnalyticaIColum n:FluopliaseRP(ICeystoneScientific),2.1 mm x SO mm, 5p (P/N: 82505-052130) 6.2 Temperature: 30*C 6.3 Mobile Phase (A ): 2 mM Ammonium Acetate in Water 6.4 Mobile Phase (B ): Methanol 6.5 Gradient Program: Tima /m ini 0.0 1.0 8.0 20.0 22.5 65 65 25 25 65 SUI 35 35 75 75 35 Flow Rate Im L/m inl 0.3 0.3 0.3 0.3 0.3 6.6 Iigeetion Volume: 1S pL (can b e increased to as m uch aa 50 pL). 6.7 Quantitation: Peak A rea-ex tern al standard calibration curve. 6.8 RunTim e: - 2 3 minutes. The above conditions are intended aa a guide and may be changed in order to optimize the HPLC system. 7.0 MS/MS System 7.1 Mode: Blectrospray Negative MRM mode, monitoring 413 --369 m/z for PFOA. The above conditions lie intended as a guide and may be changed in order to optimize die MSMS system. 8.0 Preparation o f Solutions 8.1 Mobile Phase 8.1.1 2 mM ammomum acetate in water is prepared by adding 0.154 g o f ammonium acetate to 1000 mL o f water. Alternate vohunea may b e prepared. Pag* 3 o f Page 61 o f65 Exygen Research Page 139 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 BxygeaRe m re li Method Nuraber V00017I6 I .... ........................ANALYTICAL M ETH O D Method o f Analyst for the Determination o f Perfluorooctanoic Acid (PFOA) in Small Mammal Seram by LC/MS/MS 9.0 Standard Preparation 9.1 Standard Stock/Fortificatioa Solution 9.1.1 Prepare a stock solution of~IOO pg/mL o f PFOA by weighing 10 mg o f analytical standard (corrected for purity) and dilute to 100 mL with methanol in a 125-raL LDPE bottle. 9.1.2 A 1.0 p ^ m L fortification eolution o f PFOA is prepared by bringing 1 mL o f the 100 pg/mL eolution to a final volume o f 100 with methtnol in a 12$ m L LDPB bottle. 9.1.3 A 0.1 pg/mL fortification solution o f PFOA is prepared by bringing 10 mL o f the 1.0 pg/mL solution to a final volume o f 100 with methanol in a 12$ m L LDPE bottle. 9.1.4 The stock and fortification solutions are to be stored in a refrigerator at approximately 4*C and are stable for a maximum period o f 6 months from the date o f preparation. 9.2 Standard Calibration Solutions 9.2.1 9.2.2 LC/MS/MS calibration stmidards are prepared in methanol via dilution o f die 0.1 pg/mL fortification solution. The following is a typical example: additional concentrations may be prepared as needed. Conoenhation Fins) o f Fortification Volume Diluted to Concentration Solution faa/mL) 100 100 too $.0 2.0 1.0 (mL) $.0 2.0 1.0 10 10 10 (mL) 100 100 100 too 100 too (nc/mL) 5.0 2.0 1.0 o.s 0.2 0.1 9.2.3 Store all calibration standards in 125-mL LDPE narrow-mouth bottles at 2#C to 6*C, up to six months. 9.2.4 Alternate volumes and concentrations o f standards may be prepared as needed. 10.0 Batch Set Up 10.1 Each batch o f samples extracted (typically 20 o r laaa) roust include at least one untreated control and two untreated controls fortified at known concentrations (lab control spike) to verify procedural recovery for the batch. 10.2 Requirements for field and laboratory duplicates and spikes will be specified in the quality assurance plan for tills project Page 4 o f7 Page 62 o f 65 Exygen Research Page 140 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Bxygen Protocol Number: P0000760 Ey|t> Rtmrch Method Number V000176 | ANALYTICAL METHOD Method o f Analysts for the Determination o f Perfluorooctenoic Acid (PFOA) in Small Mammal Seram by LC/MS/MS 11.0 Sample Extraction 11.1 Measure 1 mL o f u m p le into a SO m L polypropylene centrifoge tubes (fortify as needed, replace lid and mix well). Note that alternate volumes o f serum may be measured depending on the sample size available for use. 11.2 Add water to the sample for a final volume o f 20 mL. Cap tightly 11.3 Vortex fo r~ l minute. 11.4 Transfer 1 m L o f the sample using a disposable pipette into a 13 mL disposable centrifoge tube. 11.5 Add 5 m L o f acetonitrile end shake for **20 m inutes on a wrist-action shaker. 11.6 Centrifoge the tubes at -3 0 0 0 rpm for - 5 minutes. 11.7 Decant die supernatant into a 30 m L disposable centrifoge tube and add 35 mL o f water. 11.8 Condition the C u SPE cartridges (1 g, 6 m L) by passing 10 mL methanol followed by 3 mL o f HPLC wafer (~ 2 drop/sec). Do not let column ran dry 11.9 Load the se n a te on conditioned C u SPE cartridge. Discard eluate. 11.10 Elute with - 2 m L o f methanol. Collect 2 mL o f eluate into a graduated 13 m L polypropylene centrifoge tube (final volume * 2 mL). 11.11 Analyze samples using olectroapray LC/MS/MS. 12.0 Chromatography 12.1 Iqject the same amount o f each standard, sample ami fortified sample into the LC/MS/MS system. A calibration standard must precede and follow all analyzed samples. 12.2 Standard o f PFOA c o n -p o n d in g to at least five o r m ore concentration levels must be included in an analytical a ct 12.3 An entire set o f calibration standards must be included at the beginning and at foe end o f a sample s e t Standard! must be interspersed between every 3*10 samples. As an alternative, an entire set o f calibration standards may be injected at the baginning o f a set followed by calibration standards interspersed every 5-10 samples (to account for a second set o f standards), in either case, calibration standards must be the first and last injection in a sample set. 12.4 Use tin e standard curves fix quantitation. Linear standard curves are generated for die analyte by linear regression using 1/x weighting o f peak area versus calibration standard concentration using M assLynx 3.3 (or equivalent) software system. 12.5 Sample response should not exceed standard responses. Any samples that exceed standard responses should be further diluted end reanalyzed. P*Se$of? Page 63 o f 65 Exygen Research Page 141 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 ExyBmRMMrch Method Number VOOO1786 I ANALYTICAL M ETHOD Method o f A n a ly st fix the Determination o f Perfiuorooctinoic Acid (PFOA) in Small Mammal Seram by LC/MS/MS 13.0 Acceptance Criteria 13.1 Chromatogram must show a peak o f a daughter ion at 369 amu from a parent o f 413 amu. The 413 amu parent corresponds to 'th e PFOA anion, while the daughter Ion (369 amu) represents die lost o f carbon dioxide. 13.2 Method blanks must not contain PFOA at levels greater than the LOQ. If a blank contains PFOA at levels greater than 10 ng/mL, then a new blank sample must be obtained and the entire set must be re-extracted. 13.3 Recoveries o f control spikes and matrix spikes m ust be between 70-130% o f their known values. I f a control spike falls outside die acceptable limits, the entire set o f samples should be re-extracted. Any matrix spike outside 70 130% should be evaluated by die analyst to determine if re-extraction is warranted. 13.4 Any calibration standard found to b e a statistical outlier by using the Huge Error Test, may be excluded from the calculation o f the calibration curve. However, die total number o f calibration standards diet could be excluded must not exceed 20% o f the total number o f standards injected. 13.5 The correlation coefficient (R) fix calibration curves generated must be 20.992 (RJ 20.915). I f calibration results fall outside these limits, then appropriate steps must be taken to adjust instrument operation, and the standards or the relevant set o f samples should be reanalyzed. 13.6 Retention times between standards and samples must not drift more than 4 % within an analytical run. I f retention time drift exceeds this limit within an analytical ran then the set must be reanalyzed. 14.0 Calculations 14.1 Use the following equation to calculate the amount o f PFOA found (in ng/mL, baaed on peak area) using die standard curve (linear regression parameters) generated by the Mass Lynx aoftw ve program: PFOA found (ng/mL) - (Peak area - intercept1) x D F x aliquot factor slope DF factor by which die final volume was diluted, if necessary. Aliquot factor 20 14.2 For samples fortified with known amounts o f PFOA prior to extraction, use the fbllowing equation to calculate the percent recovery. Recovery (%) - [total analyte found (ng/mL) - analyte found in control (ng/mL)] analyte added (ng/mL) Pag 6 of f Page 64 o f 65 Exygen Research Page 142 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 Exygen Protocol Number: P0000760 Exygca RmmicIi Method Nunber V000I7S6 | ANA l VTICAL M ETHOD Method o f Analysis for the Detennination o f Perfluoroocunoic Acid (PFOA) in Smell Mammal Serum by LC/MS/MS 14.3 Um the following equation to convert die amount o f PFOA found in ng/mL io ppb. PUPA found ( f f h - fPPOA found (nahrdA final volume fm U l sample volume (mL) Exygen Research Page 7 of? Page 65 o f 65 Page 143 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 n RESEARCH 3058 Research Driv Phone:814-272-1039 State College, PA 16801 Fax: 814-231-1580 PRQTQCQLAMENDMENT Exygen Study Number Amendment Number. 1 Effective Date: 01/19105 P0000760______ Client Study Number Page 1 of1 ..... DESCRIPTION OF AMENDED SECTION 1) Analytical Procedure Summary V0001780:SectIon 9.1 2) Veriflcation ofAnalytical Procedure None ~ . AMENDEDTO 1) Add to Section 9.1: Section 9.1.8, Alternate weights ofstandards may be used to prepare alternate concentrations of stock solutions as necessary. Alternate levels of fortification solutions may also be prepared. 2) Low and high spiking levels ofthe analytes foreach matrix may be altered depending on samplesize available for extraction and/orto coveranalyte concentrations expected in the samples. RATIONALE 1) higher concentrations ofstandards need to be prepared in orderto spflre the sample bottles at higher levels. 2) The sample size available for smal mammal liver and serum was smaler than expected. Spiking at the pre-determined levels in the protocol puts the spiked concentration lowerthan the detection Bmit Also, the analyte levels In the ground water samples are expected to greatly exceed the pro-determined spiking levels listed in the protocol. When the levels in the samples greatly exceed the spldng levels, an accurate recovery value cannot be calculated forthe QC sample. Higher spiking levels In the bottles will coverthe analyte concentrations expected In the water samples. IMPACT ON STUDY The LOQ is 100 ng/9 for a 0.1 g sample ofsmal mammal Overand Is 1000 ngAnL for a 0.01 mL sample ofsmall mammal serum. Higher levels ofspiking for the water samplesw ll ensure that more QC recovery data can be used. J LIBRARY ID: W 0 0 0 1 2 2 6 4 Exygen Research Exygen QAU Review U S m lai/n r ADMINISTRATIVE FORM Page 144 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 txygen % / C^RESEARCH 3058 Research Drive Phone: 814-272-1039 State College, PA16801 Fax: 814-231-1580 Amendment Number. Effective Date: Exygen Study Number PROTOCOL AMENDMENT 2 03/07/05 P0000760 Client Study Number Page 1of 1 None DESCRIPTION OF AMENDED SECTION Report, page 11 of65 AMENPEPTP Instead ofone final report, Interim reports w ll be issued. RATIONALE Due to the excessive sizes of the data sets, interim reports will be issued to allow the client to receive data in a timelier manor. IMPACTONSTVPY The client will be able to receive and review the data more quickly. Studyr DiIraedW?SSiggnnaature / alM ___ N ifcipal Investigator Signature J J 6 _______________ Study Diradar Mamgamant Signatura Exyganl Sponsor SignedTM (li required) <.y LIBRARY ID: V0001226-8 $ A If Data Data 8 -M & -- S Data _^L ExygerrQAU Review 3 S eaftrliS ADM INISTRATIVE FORM Exygen Research Page 145 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 IM M IM M illa Fra-CSTM SOLUTIOMS M IT F.M2/M2 M M B p t. 3081 taeard iM ve Phot* 14-272-1039 $uce College, PA1801 Fac 14-231-1580 Amendment Number E__ff_e_c_tiv_e__D_a_t_e: Exygen Study Number PROTOCOLAMENDMENT 3 04/13106 - E tto tontStudyNumber 1flM JUL. J- ] 3 . V ; 7 i J I V ^ i * i - T > - *1 1- *11.*. r AmtyticeiProcedura Suneraey: V 000l780iM eB iodofA M iW atoriheD et^ Pa fluorooootanoteAdd (PFOA) In WaterfcyLOM3/MS." Section# 8 J end 11.0 oflhe Section 9.2. Nor>e**oedceeiredm etah*dem eybeprapeiedln orderloquenfltBie empiee directlyInjected ontotheUGMSMSayetom. ^^ Section n .a Semptoe may be anafcoed wfflm* going through the erection procedura. Instead. they mey be diutod, f appropriale, and dtrectiy rijected onto the ICMSMS systemforarariyefe. ... and fltid apfcee era not In agreement andtor were unueeWe. Dfredfy Injecting lite exfrecte may add nm e maight into the matte effceto onthe date raaulte. " " l*ff*CT ON STUDY More ueabiedata raaultemaybe obtained. m 's f BET 2 -JH - 0 ? w Bggen QAU W m UBftAKVto!V000122B4 RECEIVED T IM E JU N . 2 . 9:29R M P R IN T T IM E JMNJSTNATTVEFORM JUN. 2 , 9 :3 a m Exygen Research Page 146 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 JUL.25.2005 8:56AM EXYGEN RESEARCH NO.774 P.4 IT RESEARCH 3058 Research Drive Phone: 814-272-1039 State College, PA 16801 Fax: 814 231-1580 Amendment Number Effective Date: Exygen Study Number PROTOCOLAMENDMENT 4 07/18/06 . Ptu... ClientStudyNumber Pagel of 1 ..... NA.... DESCRIPTION F AMENDED SECTION Verificati! of Analytical Procedure, page 10 of protocol. AMENDED TO The field dupOcata can be used for the laboratory spikes and replicate when the primary sample volume Is limited. fw rw m s ,_ The sample size for a water sample ls 20 0m L If a sample site requires re-extraction for any reason, there would not be enough of the primary sample to ra p e two laboratory spikes and a replicate. The field duplicate Is technically the same sample as foe primary sample and therefore, can be used for laboratory spikes and replicates as needed- IM P A C T Q N .,m 8 B .Y ", , _ __ No negative impact on the study. Using the duplicate sample allows for the te l QC of the sample sits to be completed. Principal Invaatigator Signature U Exygen Sponsor S ignaturefif required) m `-- P a le 2 i 'J(jL JS Date ? A s / q T _____ IS * ' Exygen QAU Review /h i.'1*-- LIBR A RY ID -. V 000122S -6 ADM INISTRATIVE FORM Exygen Research Page 147 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 3058 Research Drive Phone: 814-272-1039 State College, PA 16801 Fax: 814-231-1580 Amendment Number Effective Date: Exygen Study Number PROTOCOL AMENDMENT 5 09/13/05 P760 Client Study Number: Page 1 of 1 NA . DESCRIPTION OF AMENDED SECTION A nalytical Procedure Sum m ary: V0001780:"M ethod o f A nalysis fo r the D eterm ination o f P erfluoroooctanoic A d d (PFO A) in W ater by LC/M S/M S." AMENDED TO The reanalysis o f tw o w ater sam ples, DF09-G W -131R-0-040126 and DFB-G W -135L-0040121 w ill be perform ed using the follow ing procedure: An aliqu ot o f the w ater sam ple is spiked w ith the surrogate standard a n d/or oth er m atrix spike com pounds as appropriate. Add m ethanol to the fo rtifie d aqueous sam ple. Mix and aliqu ot fo r LC/M S/M S analysis. The LC conditions include the use o f a B etasil C18, 2.1 x 50 mm colum n, a gradient o f 30% B to 100% B in 10 m inutes, w here A=2m M am m onium acetate in w a te r and B=M ethanol (6 m in eq uilibration). Use a 5 pL injection volum e and a 0.3 m L/m in flow rate. LC conditions m ay be fu rth e r m odified as appropriate fo r the system being used. An interim report w ill be issued tha t w ill indude the exact details o f the reanalysis. R A T IO N A LE DF09-G W -131R-0-040126 and DF8b-GW -135L-0-040121 could not be reported quantitatively in interim report #3 due to quality control sam ple fa ilu re s. The study director and sponsor have requested reanalysis using the above conditions. IMPACT ON STUDY No negative im pact on the study. M ore usable data m ay be obtained. Study Director Signature Principal Investigator Signature ill Signature Exygen M anagem ent Signature Sponsor Signature (if required) D ate D ate D ate D ate D ate Exygen QAU R eview V ^-T LIBRARY ID: V0001226-8 ADMINISTRATIVE FORM Exygen Research Page 148 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 CHEH EHSR 236 1B J 'C W M U C *I t .... 651 J733 1958 EP.13.2a05 1:51PM OCTOENRESDWW 09/14 '05 09:27 NO.915 02/02 Exvsen 3058 Research Drive Phone 814-272*1039 State College, P416501 F 814-231-1586 RESEARCH Amendment Number Effective Date; . Exygen Study Number PROTOCOLAMENDMENT 5 09/13/5 "P760~ ClientStudy Number. P a g a io M NA ] 3 4 U t l -u i > r J S 3 ^ p 7 A 5 X ' : ^ i T AnalyticalPrecedureSummaiy: VOO(H780rMethodorAra|yAfbrtheDteenninstioncr PerfluorooocfanoieAdd (PFOA) inWater by LC/MSiMS.' : AMENDEDtO The reanatysis of two water samples, DFCS-GW*l81RtW)40l28 and DF8*GW-i36L-0- 040121 will be performed usingmefotewing procedure: __ An aliquot ofthe watersample is spacedwiih the surrogate standard andforothermabix spike compounds se appropriate. Add methanol to the Raffled aqueous sample.- Mix and aHqiiotfor LC/MS/MS analysis. The LCcondtRona includethe use ofa Betasfl Cl8, 2.1 x 50 mm column, a gradient 6f 30% B to 100% B in 10 minutes, where A2mM ammanium aoetate mwater and 8=Methanol (6 min equilibration). Usaa5yUnjecBon volume end a 0-3 mUWn flow rale. LC condKom may be further modBed as appropriate for the system being used. An interim report will be issued thatwill indude the exactdetails oftne reanalysis. !~ RATIONALE DF09-GW-131R-0-04012G and DF86-GW-135L-0-040121 could not be reported quanfltatively in interim report #3 due to quality control sample tenures. The study director and sponsor have requested reanalysis usingthe above conditions. " " IMPACTON STUDY No negative impacton the study. More usable data may be obtained. uayDbM torSlgm ftire PrincipalInvestigatorSenabie studyDaecwStenajamBriSpnautB Bqgsn Management /jM bhr eponuor D aw CM* Dab IS C i( l s O tW ' l ' Exvoen QAU Review <7.w V r s / * r LIBRARY:Vt0122e-a RECEIVED T lfE S E P .1 4 . 1 0:4 3 flM Exygen Research AOMWISTHATTVE FORM PR IN T TIM E S E P .1 4 . 1 0 !4 4 flM Page 149 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 5-2003 02:19 FR0M:3M ENU. LA 651 778 6176 SP.i3.2005 l:5lPt1 EXYGEN RESEARCH TO:*88142311580 NO.294 ? . l P :2 '2 Exygen ^JR ESEA R C H 3058 Research Drive Phone: 814-272-1039 S tate College, PA 1680t Fax: 814-231-1580 Amendment Number Effective Date: Exygen Study Number PROTOCOLAMENDMENT 5 09/13/05 P760 ClientStudyNumber Pagai of1 NA DESCRIPTION OF AMENDEDSECTION Analytical Procedure Summary. V00Q1780^Method ofAnalysis forthe Determination of PerftuoroooctanoicAcid (PFOA) InWater by LC/MS/MS.* AMENDED TO The reanalysis of two water samples, DF09-GW-131R-0-040128 and DFB-GW-13SL-0040121 will be performed using the foiowing procedure: An aliquot ofthe watersample is spiked with the surrogate standard and/or other matrix spike compounds as appropriate. Add methanol to the fortified aqueous sample. Mix and aliquot for LC/MS/MS analysis.. The LC conditions include the use ofa Betesit C18, 2.1 x 50 mm column, a gradient of 30% B to 100% B in 10 minutes, where A2mM ammonium acetate in water and B=Methanol (6 min equKbretion). Use a 5 |iL Injection volume and a 0.3 mL/min flow rate. LC conditions may be further modified as appropriate for the system being used. An interim report win be issued that will include the exactdetails oftne reanalysis. -- -------- RAT fS M A g -------------------------------------------------------- DF09-GW-131R-0-040128 and DF8b-GW-135L-0-040121 could not be reported quantitatively in interim report #3 due to quafty control sample fafcres. The study directorand sponsorhave requested reanalysis using the above conditions. IMPACT ON STUDY No negativa snpaet on the study. More usable date maybeobtained. Study Dii m m! ; Dale / 11 Prfncal Investigator Signature Study Director Management Signature Btygen Mgnagsment Stonatura SponsorSignatura (Kraquirad) Dato bite" Dato Date Exygen QAU Review (T.** V t l / e LIBRARY ID: V 0 0 0 1 2 2 M RECEIVED TIM E S E P .1 4 . 10:3?P M Exygen Research ADMINISTRATIVE FORM P R IN T TIM E S E P .1 4 . 10:38P M Page 150 of 151 Interim Report #22 - Analysis of Soil Samples Exygen Study No.: P0000760 E RESEARCH 3058 Research Drive Phone: 814-272-1039 State College, PA 16801 Fax: 814-231-1580 Amendment Number Effective Date: Exygen Study Number PROTOCOL AMENDMENT 6 09/20/05 P760 Client Study Number Page 1 of 1 NA LIBRARY ID: V0001226-8 Exygen Research ADMINISTRATIVE FORM Page 151 of 151