Document E6G94pVmGYv3OGb67637bERn

GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 GLP10-01-02: Interim Report 24 - Analysis o f PFBS, PFHS, and PFOS in Groundwater Samples Collected at the Former Sludge Incorporation Area (FSIA) located in Decatur, AL. December 2011 Study Title Analysis of Perfluorooctane Sulfonate (PFOS), Perfluorohexane Sulfonate (PFHS) and Perfluorobutane Sulfonate (PFBS) In Groundwater, Soil and Sediment for the 3M Decatur Phase 3 . Site-Related Monitoring Program Data Requirement EPA TSCA Good Laboratory Practice Standards 40 CFR Part 792 Study Director Jaisimha Kesari P.E., DEE Weston Solutions, Inc. 1400 W eston Way W est Chester, PA 19380 Phone: 610-701-3761 Author Susan W olf 3M Environmental Laboratory Interim Report Completion Date Date of signing Performing Laboratory 3M Environmental Health and Safety Operations Environmental Laboratory 3M Center, Bldg 260-05-N-17 St. Paul, MN 55144 Project Identification GLP10-01-02-24 Total Number o f Pages 110 The testing reported herein meet the requirements of ISO/IEC 17025-2005 "General Requirements for the Competence of Testing and Calibration Laboratories", in accordance with the A2LA Certificate #2052.01. Testing that complies with this International Standard also operate in accordance with ISO 9001:2000. Testing Cert #2052.01 Page 1 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 This page has been reserved for specific country requirements. Page 2 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 GLP Compliance Statement Report Title: GLP10-01-02; Interim Report 24 - Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Collected at the Former Sludge Incorporation Area (FSIA) in Decatur, AL, December 2011 Study: Analysis of Perfluorooctane Sulfonate (PFOS), Perfluorohexane Sulfonate (PFHS) and Perfluorobutane Sulfonate (PFBS) in Groundwater, Soil and Sediment for the 3M Decatur Phase 3 Site-Related Monitoring Program. This analytical phase was conducted in compliance with Toxic Substances Control Act (TSCA) Good Laboratory Practice (GLP) Standards, 40 CFR 792, with the exceptions listed below: These are environmental samples where there is no specific test substance, no specific test system and no dosing of a test system. The reference substances have not been characterized under the GLPs and the stability under storage conditions at the test site have not been determined under GLPs. Date Page 3 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Quality A ssurance Statement Report Title: GLP10-01-02; Interim Report 24 - Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Collected at the Former Sludge Incorporation Area (FSIA) in Decatur, AL, December 2011 Study: Analysis of Perfluorooctane Sulfonate (PFOS), Perfluorohexane Sulfonate (PFHS) and Perfluorobutane Sulfonate (PFBS) in Groundwater, Soil and Sediment for the 3M Decatur Phase 3 Site-Related Monitoring Program. This analytical phase was audited by the 3M Environmental Laboratory Quality Assurance Unit (QAU), as indicated in the following table. The findings were reported to the principal investigator (P.I.), laboratory management and study director. Inspection Dates 1/16/2012-1/17/2012 Phase Data / Report Date Reported to Testing Facility Management Study Director 1/24/2012 1/24/2012 Date Page 4 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Table of Contents GLP Compliance Statement.................................................................................................................. 3 Quality Assurance Statement................................................................................................................ 4 Table of Contents...................................................................................................................................5 List of Tables......................................................................................................................................... 6 Attachments........................................................................................................................................... 7 1 Study Information............................................................................................................................ 8 2 Sum m ary.........................................................................................................................................9 3 Introduction.................................................................................................................................... 11 4 Test & Control Substances....................................................................................... ;................. 11 5 Reference Substances................................................................................................................. 12 6 Test System .................................................................................................................................. 13 7 Method Sum m ary......................................................................................................................... 13 7.1 M ethods........................................................................................................................ 13 7.2 Sample Collection..........................................................................................................13 7.3 Sample Preparation.......................................................................................................13 7.4 Analysis......................................................................................................................... 14 8 Analytical ResuIts.......................................................................................................................... 15 8.1 Calibration..................................................................................................................... 15 8.2 System Suitability..........................................................................................................15 8.3 Limit of Quantitation (LO Q )........................................................................................... 16 8.4 Continuing Calibration...................................................................................................16 8.5 Blanks............................................................................................................................ 16 8.6 Lab Control Spikes (LC Ss)........................................................................................... 16 8.7 Analytical Method Uncertainty...................................................................................... 18 Page 5 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 8.8 Field Matrix Spikes (FMS)............................................................................................ 19 9 Data Summary andDiscussion..................................................................................................... 19 10 Conclusion.....................................................................................................................................35 11 Data/Sample Retention................................................................................................................35 12 Signatures.....................................................................................................................................36 L ist of Tables Table 1. Summarized PFBS, PFHS, and PFOS Results (FSIA Groundwater, December 2011).... 10 Table 2. Sample Description Key Code............................................................................................. 13 Table 3. Instrument Parameters.........................................................................................................14 Table 4. Liquid Chromatography Conditions......................................................................................14 Table 5. Mass Transitions.................................................................................................................. 15 Table 6. Limit of Quantitation (LOQ).................................................................................................. 16 Table 7. Laboratory Control Spike Recovery..................................................................................... 17 Table 8. Analytical Uncertainty...........................................................................................................18 Table 9. Field Matrix Spike Levels..................................................................................................... 19 Table 10. DAL GW 130R 111208..................................................................................................... 20 Table 11. DAL GW 130S 111208...................................................................................................... 20 Table 12. DAL GW 130L111208...................................................................................................... 21 Table 13. DAL GW 131R 111214..................................................................................................... 21 Table 14. DAL GW 131S 111214...................................................................................................... 22 Table 15. DAL GW 131L 111214...................................................................................................... 22 Table 16. DAL GW 133R 111214......................................... 23 Table 17. DAL GW 133S 111214...................................................................................................... 23 Table 18. DAL GW 133L 111214...................................................................................................... 24 Table 19. DAL GW 13 4R 111209....................................................................................................24 Table 20. DALGW 134S 111209......................................................................................................25 Page 6 of 110 7 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Table 21. DAL GW 134L111209.....................................................................................................25 Table 22. DAL GW 135R 111214................................................................................................... 26 Table 23. DAL GW 135S 111214................................................................................................... 26 Table 24. DAL GW 135L 111214................................................................................................... 27 Table 25. DAL GW 136S 111213................................................................................................... 27 Table 26. DAL GW 136L 111213................................................................................................... 28 Table 27. DAL GW 137S 111209................................................................................................... 28 Table 28. DAL GW 137L 111209................................................................................................... 29 Table 29. DAL GW 138R 111214................................................................................................... 29 Table 30. DAL GW 138S 111214................................................................................................... 30 Table 31. DAL GW 138L 111214....................................................................................................30 Table 32. DAL GW 140R 111213...................................................................................................... 31 Table 33. DAL GW 141R 111214...................................................................................................... 31 Table 34. DAL GW 142R 111213...................................................................................................... 32 Table 35. DAL GW 143R 111209...................................................................................................... 32 Table 36. DAL GW 144R 111213................................ !................................................................... 33 Table 37. Trip Blank 1 ........................................................................................................................ 33 Table 38. Rinseate Blanks.................................................................................................................34 Table 39. Surrogate Recovery........................................................................................................... 34 Attachments Attachment A: Protocol Amendment 24.............................................................................................37 Attachment B: Representative Chromatograms and Calibration C urves....................................... 45 Attachment C: Analytical Method ETS-8-044.1............................................................................... 86 Attachment D: Method Deviation.....................................................................................................109 Page 7 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 1 Study Information Sponsor 3M Company Sponsor Representative Gary Hohenstein 3M EHS Operations 3M Building 224-5W-03 Saint Paul, MN 55144-1000 Phone: (651) 737-3570 Study Director Jaisimha Kesari, P.E., DEE Weston Solutions, Inc. West Chester, PA 19380 Phone: (610) 701-3761 Fax: (610)701-7401 j.kesari@ w estonsolutions.com Study Location Testing Facility 3M EHS Operations 3M Environmental Laboratory Building 260-5N-17 St. Paul, MN 55144 Study Personnel W illiam K. Reagen, Ph.D., 3M Laboratory Manager Cleston Lange, Ph.D., Principal Analytical Investigator, (clanqe@mmm.com): phone (651)-733-9860 Susan Wolf, 3M Analyst Chelsie Grochow, Analyst Kelly Ukes, Analyst Study Dates Study Initiation: March 8, 2010 Interim 24 Experimental Termination: January 5,2012 Interim Report Completion: Date of Interim Report Signing Location of Archives All original raw data and the analytical report have been archived at the 3M Environmental Laboratory according to 40 CFR Part 792. The test substance and analytical reference standard reserve samples are archived at the 3M Environmental Laboratory according to 40 CFR Part 792 Page 8 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 2 Summary The 3M Environmental Laboratory received groundwater samples from wells located at the Former Sludge Incorporation Area (FSIA) in Decatur, AL, representing twenty-seven different sampling locations. A total of eighty-eight sample bottles were received at the 3M Environmental Laboratory for perfluorooctane sulfonate (PFOS), perfluorohexane sulfonate (PFHS) and perfluorobutane sulfonate (PFBS), and included duplicate groundwater samples from each sampling location. Samples also included a field matrix spike (FMS) sample for each location, a trip blank set containing M illi-QTM water and appropriate trip blank spikes, and two equipment rinseate blanks. The equipment rinseate blanks did not have FMS samples prepared for determination of PFBS, PFHS, and PFOS recovery. The groundwater samples, trip blanks and two equipment rinseate blanks associated with GLP10-0102-24 were received from Weston personnel on December 20, 2011. All of the samples were prepared and analyzed for PFBS, PFHS, and PFOS following 3M Environmental Laboratory Method ETS-8044.1 and conducted under 3M project GLP-10-01 -02-24. Many of the groundwater samples required dilution to attain PFBS, PFHS, and/or PFOS concentrations within the range of the curve, in some instances up to 100-fold dilution were required. The average measured PFBS, PFHS, and PFOS concentrations are summarized in Table 1. The equipment rinseate blanks and the trip blank sample were below the lower limit of quantitation (LLOQ) for PFBS, PFHS, and PFOS, indicating adequate control of sample contamination during shipping and sample collections. The PFBS concentration results for all groundwater samples ranged from <0.0250 ng/mL to 64.4 ng/mL. The PFHS concentration results for all groundwater samples ranged from 0.0594 ng/mL to 827 ng/mL. The PFOS concentration results for all groundwater samples ranged from 0.420 ng/mL to 1570 ng/mL. The analytical method uncertainties associated with the reported results are: PFBS + 17%, PFHS + 18% and PFOS + 19%. For sample DAL GW 134L, the method uncertainty was expanded for PFOS. Page 9 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Table 1. Summarized PFBS, PFHS, and PFOS Results (FSIA Groundwater, December 2011). Sampling Location DALGW 130R 111208 DALGW 130S 111208 DAL GW 130L 111208 DAL GW 131R 111214 DAL GW 131S 111214 DAL GW 131L 111214 DAL GW 133R 111214 DALGW 133S 111214 DAL GW 133L 111214 DALGW 134R 111209 DALGW 134S 111209 DAL GW 134L 111209 DALGW 135R 111214 DALGW 135S 111214 DAL GW 135L 111214 DALGW 136S 111213 DAL GW 136L 111213 DALGW 137S 111209 DAL GW 137L 111209 DALGW 138R 111214 DALGW 138S 111214 DALGW 138L 111214 DALGW 140R 111213 DAL GW 141R 111214 DALGW 142R 111213 DALGW 143R 111209 DALGW 144R 111213 Trip Blanks (M illi-QTM Water) DALGW 140R RB 111213 D ALG W 133LR B 111214 PFBS Avg. Cone. (ng/mL) RPD 15.1 6.6% 3.27 1.8% 1.49 2.0% 36.6 0.55% 0.688 0.15% 2.86 1.1% 2.94 15% 11.3 0.89% 7.90 2.4% 47.5 0.21% 0.364 1.6% 10.5 1.9% 64.4 1.6% 0.991 6.0% 7.09 0.85% 1.77 3.4% 1.91 2.6% 0.120 13% 0.0868 14% 7.40 0.95% 3.64 4.1% <0.0250 1.27 6.3% 0.0342 5.3% 1.77 7.4% 0.715 1.7% 2.02 3.0% <0.0250 <0.0250 <0.0250 PFHS Avg. Cone. (ng/mL) RPD 119 0.84% 25.6 1.6% 11.7 5.1% 182 2.2% 4.78 3.6% 18.8 0.53% 20.9 17% 79.7 1.4% 58.7 4.1% 561 0.54% 4.22 1.7% 109 0.0% 827 1.6% 7.90 2.9% 47.6 1.5% 11.4 3.5% 12.3 0.82% 0.113 21% (2> 0.0992 9.8% 35.3 1.4% 17.8 1.7% 0.0594 12% 14.1 2.1% 0.254 4.3% 9.97 4.6% 2.33 0.0% 7.54 0.66% <0.0250 <0.0250 <0.0250 PFOS Avg. Cone. (ng/mL) RPD 467 9.9% 90.8 10% 66.3 1.4% 1370 1.5% 4.56 3.7% 27.9 0.72% 39.1 10% 174 2.9% 252 2.8% 1570 1.3% 17.5 0.0% 436 1.6% <1) 1110 4.5% 5.81 7.6% 33.8 7.7% 59.4 1.7% 64.4 3.0% 0.517 32% (23) 1.62 8.0% 12(3)4 300 0.0% 160 1.3% 0.420 20% (3) 102 3.0% 4.87 4.1% (3'4) 94.4 0.32% 15.6 6.4% 26.7 1.9% (3) <0.0371 <0.0371 <0.0371 All samples reported using external standard calibration unless noted otherwise. The analytical method uncertainties associated with the reported results are: PFBS 17%, PFHS 18%, and PFOS 19%. (1) The analytical uncertainty has been adjusted for PFOS based on FMS recovery for DAL GW 134L to 39%. (2) The RPD did not meet method acceptance criteria of <20%. (3) Sample results reported using internal standard calibration. (4) Sampling location did not have an appropriate FMS spike level to sufficiently evaluate PFOS recovery. Page 10 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS In Groundwater Decatur, AL - December 2011 3 Introduction This analytical study was conducted as part of the Phase 3 Environmental Monitoring and Assessment Program for the 3M facility located in Decatur, Alabama. The objective of the overall program is to gain information regarding concentrations of perfluorooctane sulfonate (PFOS), perfluorohexane sulfonate (PFHS) and perfluorobutane sulfonate (PFBS), in various environmental media such as groundwater, soils and sediments that are associated with and near the Decatur facility. This analytical study was conducted to analyze groundwater samples collected from various wells located at the Former Sludge Incorporation Area (FSIA) in Decatur, AL for PFBS, PFHS, and PFOS in an effort to characterize on site groundwater conditions. The 3M Environmental Laboratory prepared sample containers (250 mL high-density polyethylene bottles) which were shipped to Decatur, AL Weston personnel prior to field sampling. Sample containers for each sampling location included a field sample, field sample duplicate, and a field matrix spike sample. Each empty container was marked with a "fill to here" line to produce a final sample volume of 200 mL. Containers designated for field matrix samples were fortified with an appropriate matrix spike solution containing PFBS (linear), PFHS (linear), and PFOS (linear) prior to being sent to the field for sample collection. See section 8.8 of the report for field matrix spike levels. Sample bottles for select sampling locations were fortified with internal standard and surrogate prior to sample collection to aid in sample analysis. Where applicable, internal standards were used to aid in the data quality objectives. Sampling locations reported for PFOS using internal standard calibraton included: DAL GW 137S, DAL GW 137L, DAL GW 138L, DAL GW 141R, and DAL GW 144R. Samples were prepared and analyzed according to the procedure defined in 3M Environmental Laboratory method ETS-8-044.1 "Method of Analysis for the Determination of Perfluorinated Compounds In W ater by High Performance Liquid Chromatography/Mass Spectrometry Direct Injection Analysis". Table 1 summarizes the average PFBS, PFHS, and PFOS concentrations for the duplicate samples collected, the trip blank, and the aqueous equipment rinseate blank samples. Tables 10-38 summarize the individual sample results and the associated field matrix spike recoveries. Surrogate recoveries for samples quantitated using internal standard calibration are summarized in Table 39. All results for the quality control samples prepared and analyzed with the samples are reported and discussed elsewhere in this report. 4 Test & Control Substances There was not a test substance or control substances in the classic sense of a GLP study. This study was purely analytical in nature. 1 I\ Page 11 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 i 5 Reference Substances R eference Substance Chemical Name Chemical Formula Identifier Use Source Expiration Date Storage Conditions Chemical Lot Number TCR Number Physical Description Purity PFBS (predom inantly lin ear isom er) Perfluorobutane sulfonate c 4f9s o 3k + Potassium Salt Target Analyte Reference Standard 3M 1/10/2017 Frozen 101 TCR-121 White Powder 96.7% PFHS (lin ea r isom er) Perfluorohexane sulfonate C8F13SO3 Na Sodium Salt Target Analyte Reference Standard Wellington 3/25/2018 Frozen LPFHxSAM08 TCR08-0018 Crystalline 100% PFOS (branched and lin ear isom ers) Perfluorooctane sulfonate C8F17SO3 K* Potassium Salt CAS # br-PFOSK Target Analyte Reference Standard Wellington 3/17/2014 Frozen brPFOSK0708 TCR11-0010 Liquid N A(1) R eference Substance Chemical Name Chemical Formula Identifier Use Source Expiration Date Storage Conditions Chemical Lot Number TCR Number Physical Description Purity PFOS (branched and lin ear isom ers) Perfluorooctane sulfonate C8F17SO3 K+ Potassium Salt CAS #2795-39-3 FMS Reference Standard Aldrich 2/4/2014 Room Temperature 1424328V TCR11-0028 White Powder 99.7% 13C,,-PFOS Surrogate Perfluorooctane sulfonate C8F17S 0 3'Na+ Sodium Salt Surrogate Recovery Standard Wellington 9/8/2013 Frozen MPFOS0910 TCR10-0044 Liquid N A<1) 13Cs-PFOS Sodium Perfluorooctanesulfbnate 13C8F17S 0 3'Na+ CAS # MPFC-C-0511 Internal Standard Wellington 05/25/2014 Frozen 052411 TCR11-0016 Liquid n a (2) (1) Reference standard at a concentration of 50 2.5 pg/mL (2) Custom mixture o f seven mass-labeled (,3C) perfluoroalkylcaiboxylic adds, two mass labeled (13C) perfiuoroalkylsulfonates and one mass-labeled ( '3C) perfluoro-1-octanesulfonamide at a nominal concentration of 5 pg/mL Page 12 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 6 Test System The test systems for this study are groundwater samples collected from wells located in Decatur, AL by Weston Solutions, Inc. personnel. Samples for this study are "real world" samples, not dosed with a specific lot of test substance. Table 2. Sample Description Key Code. String Number Example 1 2 3 4 5 6 String Descriptor DAL GW 131R 0111214 Sampling Location Well ID Sampling Point Well Level Sampling Date Sample Type Example DAL = Decatur, Alabama G W = Groundwater Example: 131 R = Residuum shallow water-bearing zone L = Bedrock water-bearing zone S = Epikarst middle water-bearing zone 1 1 1 2 1 4 -December 14,2011 0=primary sample DB=duplicate sample FMS = Field Matrix Spike 7 Method Summary 7.1 Methods Analysis for all analytes was completed following 3M Environmental Laboratory method ETS-8-044.1 "Method of Analysis for the Determination of Perfluorinated Compounds In W ater by High Performance Liquid Chromatography/Mass Spectrometry Direct Injection Analysis". 7.2 Sample Collection Samples were collected in 250 mL NalgeneTM (high-density polyethylene) bottles prepared at the 3M Environmental Laboratory. Sample bottles associated with GLP10-01-02-24 were returned to the laboratory at ambient conditions on December 20, 2011. Samples were stored refrigerated at the laboratory after receipt. A set of laboratory prepared Trip Blank and Trip Blank field matrix spikes were sent with the sample collection bottles. 7.3 Sample Preparation Samples were prepared by removing an aliquot of the well mixed sample and placing it in an autovial for analysis. Samples that required dilution were prepared by dilutions 1mL sample with 9 mLs of Milli-Q water (dilution 1:10). Samples that required dilution were prepared by diluting 1mL of sample with 9 mLs of Milli Q water (dilution 1:10). Samples requiring a 1:100 dilution were prepared by diluting 0.1 mL of sample with 9.9 mLs of MilliQ water Sampling locations reported for PFOS using internal standard calibraton: During the preparation of the laboratory control samples, an aliquot of a separate internal standard spiking solution was added to the laboratory control samples (nominal concentration of 1 ng/mL). The samples bottles were spiked with an internal standard mix at a nominal concentration of 1 ng/mL prior to being sent to the field for sample collection. Page 13 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 7.4 Analysis All study samples and quality control samples were analyzed for PFBS, PFHS, and PFOS using high performance liquid chromatography/ tandem mass spectrometry (HPLC/MS/MS). Detailed instrument parameters, the liquid chromatography gradient program, and the specific mass transitions analyzed are described in the raw data hard copies placed in the final data packet, and are briefly described below. Table 3. Instrument Parameters. Instrument Name Analytical Method Followed Analysis Date Liquid Chromatograph Guard column Analytical column Injection Volume Mass Spectrometer Ion Source Electrode Polarity Software ETSKirk ETS-8-044.1 12/27/11 - PFBS, PFHS, and PFOS External standard calibration Agilent 1290 Betasil C18 (4.6 mm X 100 mm), 5u Betasil C18 (4.6 mm X 100 mm), 5u 10 nL Applied Blosystems API 5500 Turbo Spray Turbo ion electrode Negative Analyst 1.5.2 ETS Buster ETS-8-044.1 1/4/12 - PFOS Internal standard calibration Agilent 1100 Prism RP (2.1 mm X 50 mm), 5n Betasil C18 (2.1 mm X 100 mm), 5u 10 uL Applied Biosystems API 4000 Turbo Spray Turbo ion electrode Negative Analyst 1.4.2 Table 4. Liquid Chromatography Conditions. Step Number Total Time (min) 00 1 2.0 2 14.5 3 15.5 4 16.5 5 20.0 00 1 2.0 2 14.5 3 15.5 4 16.5 5 20.0 Flow Rate (pL/min) Percent A (2 mM ammonium acetate) ETS-8-044.1 Analyzed 12/27/11 750 97.0 750 97.0 750 5.0 750 5.0 750 97.0 750 97.0 ETS-8-044.1 Analyzed 1/4/12 300 97.0 300 97.0 300 5.0 300 5.0 300 97.0 300 97.0 PercentB (Methanol) 3.0 3.0 95.0 95.0 3.0 3.0 3.0 3.0 95.0 95.0 3.0 3.0 Page 14 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Table 5. Mass Transitions. Analyte Mass Transition Q1/Q3 Reference M aterial S tru ctu re Internal Standard Mass Transition Q1/Q3 PFBS 299/80 299/99 Linear NA NA PFHS 399/80 399/99 Linear NA NA PFOS 499/80 499/99 499/130 Branched and Linear [ n C$jPFOS<1> 507/80 113C4lPFOS - surrogate m 503/80 Linear [ 13C8]PFOS <v 507/80 Dwell time was 20 or 50 msec for each transition. The individual transitions were summed to produce a "total ion chromatogram" (TIC), which was used for quantitation. (1) Internal standard and surrogate were only used for PFOS for sampling locations DAL GW 137S, DAL GW 137L, DAL GW 138L, DAL GW 141R, and DAL GW 144R. 8 Analytical Results 8.1 Calibration Samples were analyzed against an external standard calibration curve. Calibration standards were prepared by spiking known amounts of the stock solution containing the target analytes into prepared in Milli Q water. A total of fifteen spiked standards ranging from 0.025 ng/mL to 100 ng/mL (nominal) were analyzed. A quadratic, 1/x weighted, calibration curve of the peak area counts was used to fit the data for each analyte. The data were not forced through zero during the fitting process. Calculating the standard concentrations using the peak area confirmed accuracy of each curve point. For sampling locations DAL GW 137S, DAL GW 137L, DAL GW 138L, DAL GW 141R, and DAL GW 144R for PFOS: Samples were analyzed using a stable isotope internal standard calibration curve. Calibration standards were prepared by spiking known amounts of the stock solution containing the target analytes into a laboratory-prepared synthetic groundwater containing calcium and magnesium. A separate internal standard spiking solution was prepared and an aliquot was added at the same level to all calibration standards and laboratory control samples at a nominal concentration of 1 ng/mL. A calibration curve ranging from 0.025 ng/mL to 100 ng/mL (nominal) was analyzed (0.025 ng/mL to 10 ng/mL for [13C4]PFOS surrogate). A quadratic, 1/x weighted, calibration curve of the peak area ratios was used to fit the data for each analyte. The data were not forced through zero during the fitting process. Calculating the standard concentrations using the peak area ratios and the resultant calibration curve confirmed accuracy of each curve point. Each curve point was quantitated using the overall calibration curve and reviewed for accuracy. Method calibration accuracy requirements of 10025% (10030% for the lowest curve point) were met for all analytes. The correlation coefficient (r) was greater than 0.995 for all analytes. 8.2 System Suitability A calibration standard was analyzed four times at the beginning of each analytical sequence to demonstrate overall system suitability. The acceptance criteria of less than or equal to 5% relative standard deviation (RSD) for peak area and retention time criteria of less than or equal to 2% RSD was met for PFBS, PFHS, PFOS, and [13C4]PFOS surrogate. Page 15 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 8.3 Limit of Quantitation (LOQ) The LOQ for this analysis is the lowest non-zero calibration standard in the curve that meets linearity and accuracy requirements and for which the area counts or area ratio are at least twice those of the appropriate blanks. The LOQ for all analytes can be found in Table 6. Table 6. Lim it o f Quantitation (LOQ). Analysis Date 12/27/11 Analysisexternal standard calibration 1/4/12 Analysis internal standard calibration NA = Not Applicable D ilu tio n 1 10 100 1 PFBS LOQ, np/mL 0.0250 0.250 2.50 NA PFHS LOQ, np/mL 0.0250 0.250 2.50 NA PFOS LOQ, np/mL 0.0371 0.371 3.71 0.0928 8.4 Continuing Calibration During the course of each analytical sequence, continuing calibration verification samples (CCVs) were analyzed to confirm that the instrument response and the initial calibration curve were still in control. All CCVs met method criteria of 100% 25% for PFBS, PFHS, PFOS, and [13C4]PFOS surrogate. 8.5 Blanks Three types of blanks were prepared and analyzed with the samples: solvent blanks, trip blanks, and equipment rinseate blanks. Solvent blank results were reviewed and used to evaluate method performance to determine the LOQ for PFBS, PFHS, PFOS, and [13C4]PFOS surrogate. Trip blanks reflect the shipping and sample collection conditons the sample bottles and samples experience. Equipment rinseate blanks are aqueous samples that reflect the efficiency of equipment cleaning in the field between different sample collections and are proof of no cross contamination of samples from the equipment. 8.6 Lab Control Spikes (LCSs) Low, mid, and high-level lab control spikes were prepared and analyzed in triplicate with each preparation set. LCSs were prepared by spiking known amounts of the analyte into Milli Q water or synthetic groundwater to produce the desired concentration. The spiked water samples were then prepared and analyzed in the same manner as the samples. The method acceptance criteria, average of LCS at each level should be within 100% 20% with an RSD <20%, were met for PFBS, PFHS, PFOS, and [13C4]PFOS surrogate LCS samples. The following calculations were used to generate data in Table 7 for laboratory control spikes. ,LCS Percen.tnR, ecovery =C--a--l-c-u--l-a--te--d---C--o--n--c--e--n--tr-a--t-i-o--n---,*. 1. 0. 0. .% Spike Concentration standard deviation LCS replicates LCS%RSD = 100% average LCS recovery Page 16 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS In Groundwater Decatur, AL - December 2011 Table 7. Laboratory Control Spike Recovery. ETS-8-044.1 Analyzed 12/27/11 External standard calibration Lab ID Spiked Concentration (ng/mL) PFBS Calculated Concentration (ng/mL) LCS-111221-4 LCS-111221-5 LCS-111221-6 0.498 0.498 0.498 0.598 0.583 0.584 Average %RSD LCS-111221-7 LCS-111221-6 LCS-111221-9 4.98 4.98 4.98 118 1.5% 5.78 5.70 5.68 Average %RSD LCS-111221-10 LCS-111221-11 LCS-111221-12 115% 0.87% 29.9 26.3 29.9 30.2 29.9 28.0 Average %RSD 94.2% 7.0% Spiked Concentration YoRecovery (ng/mL) 120 0.497 117 0.497 117 0.497 116 4.97 115 4.97 114 4.97 87.9 29.9 101 29.9 93.8 29.9 PFHS Calculated Concentration (ng/mL) 0.506 0.496 0.488 100% 1.9% 5.03 4.98 5.07 101% 0.99% 24.4 28.9 29.4 92.1% 10% YoRecovery 102 99.9 98.2 101 100 102 81.5 96.5 98.3 ETS-8-044.1 Analyzed 12/27/11 External standard calibration PFOS 'Branched and Linear) Lab ID Spiked Calculated Concentration Concentration (ng/mL) (ng/mL) YoRecovery LCS-111221-4 LCS-111221-5 LCS-111221-6 0.462 0.462 0.462 0.395 0.383 0.377 85.6 82.9 81.7 Average %RSD 83.4% 2.4% LCS-111221-7 LCS-111221 -8 LCS-111221-9 4.62 4.62 4.62 5.18 112 5.11 111 5.06 109 Average %RSD LCS-111221-10 LCS-111221-11 LCS-111221-12 27.8 27.8 27.8 111% 1.4% 23.2 26.4 26.1 83.5 94.8 93.8 Average %RSD 90.7% 6.9% Page 17 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Table 7 continued. Laboratory Control Spike Recovery. ETS-8-044.1 Analyzed 1/4/12 Internal standard calibration Lab ID PFOS (Branched and Linear) Spiked Calculated Concentration Concentration (ng/mL) (ng/mL) %Recovery l ' 3CJPFOS surrogate Spiked Calculated Concentration Concentration (ng/mL) (ng/mL) %Recovery LCS-120104-1 LCS-120104-2 LCS-120104-3 0.184 0.184 0.184 0.214 116 0.189 0.199 108 0.189 0.209 113 0.189 0.201 0.209 0.212 106 111 112 Average %RSD LCS-120104-4 LCS-120104-5 LCS-120104-6 1.84 1.84 1.84 112 3.6% 2.14 2.05 2.08 116 111 113 110% 2.9% 1.89 2.13 1.89 2.05 1.89 2.11 113 109 112 Average %RSD LCS-120104-7 LCS-120104-8 LCS-120104-9 113% 2.2% 18.4 16.5 18.4 17.3 18.4 17.2 89.6 93.8 93.6 111% 1.9% Average %RSD 92.3% 2.6% 8.7 Analytical Method Uncertainty Analytical uncertainty is based on historical QC data that is control charted and used to evaluate method accuracy and precision. The method uncertainty is calculated following ETS-12-012.2. The standard deviation is calculated for the set of accuracy results (in %) obtained for the QC samples. The method uncertainty is calculated by multiplying the standard deviation by a factor of 2, which corresponds to a confidence level of 95%. The most recent 50 data points were used to generate the method uncertainty values listed in Table 8. Table 8. Analytical Uncertainty. Analyte PFBS PFHS PFOS Standard Deviation 8.51 8.98 9.29 Method Uncertainty 17% 18% 19% Page 18 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 8.8 Field Matrix Spikes (FMS) A field matrix spike (FMS) was collected at each sampling point to verify that the analytical method is applicable to the collected matrix. Field matrix spikes were generated by adding a measured volume of field sample to a container spiked by the laboratory with PFBS (linear), PFHS (linear), and PFOS (linear+branched) prior to shipping sample containers for sample collection. Field matrix spike recoveries within method acceptance criteria of 10030% confirm that "unknown" components in the sample matrix do not significantly interfere with the extraction and analysis of the analytes of interest. Field matrix spike concentrations must be 50% of the sample concentration to be considered an appropriate field spike. Field matrix spikes are presented in section 9 of this report. Table 9. Field Matrix Spike Levels. Sampling Location 136R, 137R, 137S, 137L, 138L, 141R 131S, 134S, 135S, 143R 130L, 142R, 144R 130S, 131L, 133R, 136S, 136L, 140R 133S, 133L, 135L, 138R, 138S 130R, 134L, CW26C, CW26L 131R, 134R, 135R Trip Blank Spike Level FMS FMS FMS FMS FMS FMS FMS Low Mid Mid-High High PFBS, ng/mL 1.01 20.2 50.5 101 202 503 2010 1.01 20.2 101 503 PFHS, ng/mL 0.998 20.0 49.9 99.8 200 499 2000 0.998 20.0 99.8 499 PFOS, ng/mL 0.998 20.0 49.9 99.8 200 499 2000 0.998 20.0 99.8 499 FMS Recovery = -(-S--a-m--p--l-e--C--o-n--c-e-n--t-r-a-t-i-o--n--o--f-F--M---S----A---v-e--r-a-g-e---C--o--n-c--e-n--tr-a--t-i-o-n---:-F--ie--l-d--S-a--m- p--l-e--&---F-i-e--ld--S--a-m--p--l-e--D--u--p.) *,, 1_0_0%_ Spike Concentraton 9 Data Summary and Discussion The tables below summarize the sample results and field matrix spike recoveries for the sampling locations as well as the Trip Blanks and rinseate blanks. Results and average values are rounded to three significant figures according to EPA rounding rules. Because of rounding, values may vary slightly from those listed in the raw data. Field matrix spike recoveries meeting the method acceptance criteria of 30%, demonstrate that the method was appropriate for the given matrix and their respective quantitative ranges. DAL GW 134L - The recovery of the FMS sample for PFOS was 61.2%. Based on the recovery of the FMS sample, the analytical uncertainty has been expanded for PFOS to 39%. DAL GW 137S - The sample/sample duplicate RPD was 21% for PFHS and 32% for PFOS. DAL GW 141R - Sampling location did not have an appropriate FMS spike level for PFOS to accurately assess recovery. Page 19 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Table 10. DAL GW 130R 111208 PFBS PFHS PFOS 3M LIM SID Description C oncentration (ng/mL) %Recovery GLP10-01-02-24-001 GLP10-01 -02-24-002 GLP10-01 -02-24-003 DAL-GW-130R-0-111208 DAL-GW-130R-DB-111208 DAL-GW-130R-FMS-111208 14.6 15.6 482 NA NA 92.8 Average Concentration (ng/mL) %RPD 15.1 ng/mL 6.6% Concentration (ng/mL) % R ecovery 118 NA 119 NA 551 86.7 119 ng/mL 0.84% Concentration (ng/mL) % R ecovery 444 NA 490 NA 878 82.4 467 ng/mL 9.9% NA = Not Applicable Samples diluted 1:100 and reported from external standard calibration. Table 11. DAL GW 130S 111208 PFBS PFHS PFOS 3M LIM SID D escriptio n Concentration (ng/mL) %Recovery GLP10-01-02-24-004 GLP10-01 -02-24-005 GLP10-01-02-24-006 DAL-GW-130S-0-111208 DAL-GW-130S-DB-111208 DAL-GW-130S-FMS-111208 3.24 3.30 97.7 NA NA 93.5 Average Concentration (ng/mL) %RPD 3.27 ng/mL 1.8% C oncentration (ng/mL) % R ecovery 25.8 NA 25.4 NA 110 84.6 25.6 ng/mL 1.6% Concentration (ng/mL) YoRecovery 86.1 NA 95.4 NA 186 95.4 90.8 ng/mL 10% NA = Not Applicable Samples diluted 1:10 and reported from external standard calibration. Page 20 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Table 12. DAL GW 130L 111208 PFBS PFHS PFOS 3MLIM SID D escriptio n Concentration (ng/mL) YoRecovery GLP10-01-02-24-007 GLP10-01-02-24-008 GLP10-01-02-24-009 DAL-GW-130L-0-111208 DAL-GW-130L-DB-111208 DAL-GW-130L-FMS-111208 1.50 1.47 52.1 NA NA 100 Average Concentration (ng/mL) i %RPD 1.49 ng/mL 2.0% C oncentration (ng/mL) YoRecovery 11.4 NA 12.0 NA 54.7 86.2 11.7 ng/mL 5.1% Concentration (ng/mL) YoRecovery 65.8 NA 66.7 NA 110 87.7 66.3 ng/mL 1.4%, NA = Not Applicable Samples diluted 1:10 and reported from external standard calibration. Table 13. DAL GW 131R 111214 PFBS PFHS PFOS 3MLIMS ID D escriptio n Concentration (ng/mL) %Recovery GLP10-01-02-24-010 GLP10-01-02-24-011 GLP10-01-02-24-012 DAL-GW-131R-0-111214 DAL-GW-131R-DB-111214 DAL-GW-131R-FMS-111214 36.5 36.7 2020 NA NA 98.7 Average Concentration (ng/mL) %RPD 36.6 ng/mL 0.55% Concentration (ng/mL) YoRecovery 184 180 1970 NA NA 89.4 182 ng/mL 2.2% Concentration (ng/mL) YoRecovery 1360 1380 2980 NA NA 80.5 1370 ng/mL 1.5%, NA = Not Applicable Samples diluted 1:100 and reported from external standard calibration. Page 21 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Table 14. DAL GW 131S 111214 PFBS PFHS PFOS 3MLIM SID D escriptio n Concentration (ng/mL) XoRecovery GLP10-01 -02-24-013 GLP10-01-02-24-014 GLP10-01-02-24-015 DAL-GW-131S-0-111214 DAL-GW-131S-DB-111214 DAL-GW-131S-FMS-111214 0.688 0.687 18.3 NA NA 87.2 Average Concentration (ng/mL) XoRPD 0.688 ng/mL 0.15% C oncentration (ng/mL) XoRecovery 4.86 NA 4.69 NA 21.9 85.6 4.78 ng/mL 3.6% Concentration (ng/mL) XoRecovery 4.64 NA 4.47 NA 20.2 78.2 4.56 ng/mL 3.7Xo NA = Not Applicable Samples diluted 1:10 and reported from external standard calibration. Table 15. DAL GW 131L 111214 PFBS PFHS PFOS 3MLIM SID D escrip tio n C oncentration (ng/mL) XoRecovery GLP10-01-02-24-016 GLP10-01-02-24-017 GLP10-01-02-24-018 DAL-GW-131L-0-111214 DAL-GW-131L-DB-111214 DAL-GW-131L-FMS-111214 2.84 2.87 95.7 NA NA 91.9 Average Concentration (ng/mL) XoRPD 2.86 ng/mL 1.1Xo C oncentration (ng/mL) XoRecovery 18.8 NA 18.7 NA 112 93.4 18.8 ng/mL 0.53% Concentration (ng/mL) XoRecovery 28.0 NA 27.8 NA 103 75.3 27.9 ng/mL 0.72% NA = Not Applicable Samples diluted 1:10 and reported from external standard calibration. Page 22 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Table 16. DAL GW 133R 111214 PFBS PFHS PFOS 3M LIM S ID D e scrip tio n Concentration (ng/mL) %Recovery GLP10-01-02-24-019 DAL-GW-133R-0-111214 3.16 NA GLP10-01-02-24-020 DAL-GW-133R-DB-111214 2.71 NA GLP10-01-02-24-021 DAL-GW-133R-FMS-111214 95.9 92.0 Average Concentration (ng/mL) %RPD 2.94 ng/mL 15% Concentration (ng/mL) %Recovery 22.7 NA 19.1 NA 112 91.3 20.9 ng/mL 17% Concentration (ng/mL) %Recovery 41.1 NA 37.1 NA 121 82.1 39.1 ng/mL 10% NA = Not Applicable Samples diluted 1:10 and reported from external standard calibration. Table 17. DAL GW 133S 111214 PFBS PFHS PFOS 3MLIM SID D escriptio n Concentration (ng/mL) % R ecovery GLP10-01-02-24-022 GLP10-01-02-24-023 GLP10-01-02-24-024 DAL-GW-133S-0-111214 DAL-GW-133S-DB-111214 DAL-GW-133S-FMS-111214 11.3 11.2 204 NA NA 95.4 Average Concentration (ng/mL) %RPD 11.3 ng/mL 0.89% C oncentration (ng/mL) %Recovery 80.2 NA 79.1 NA 278 99.2 79.7ng/mL 1.4% Concentration (ng/mL) %Recovery 176 NA 171 NA 319 72.8 174 ng/mL 2.9% NA = Not Applicable Samples diluted 1:10 and reported from external standard calibration. Page 23 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Table 18. DAL GW 133L 111214 PFBS PFHS PFOS 3MLIM SID D escriptio n Concentration (ng/mL) X R e co ve ry GLP10-01-02-24-025 GLP10-01-02-24-026 GLP10-01-02-24-027 DAL-GW-133L-0-111214 DAL-GW-133L-DB-111214 DAL-GW-133L-FMS-111214 7.80 7.99 198 NA NA 94.1 Average Concentration (ng/mL) %RPD 7.90 ng/mL 2.4% C oncentration (ng/mL) %Recovery 59.9 NA 57.5 NA 241 91.2 58.7 ng/mL 4.1% C oncentration (ng/mL) %Recovery 248 NA 255 NA 411 79.8 252 ng/mL 2.8% NA = Not Applicable Samples diluted 1:10 and reported from external standard calibration. Table 19. DAL GW 134R 111209 PFBS PFHS PFOS 3MLIM SID D e scrip tio n Concentration (ng/mL) % R ecovery GLP10-01-02-24-028 GLP10-01-02-24-029 GLP10-01-02-24-030 DAL-GW-134R-0-111209 DAL-GW-134R-DB-111209 DAL-GW-134R-FMS-111209 47.4 47.5 1870 NA NA 90.7 Average Concentration (ng/mL) %RPD 47.5 ng/mL 0.21% C oncentration (ng/mL) %Recovery 559 562 2360 NA NA 90.0 561 ng/mL 0.54% C oncentration (ng/mL) YoRecovery 1580 NA 1560 3080 NA 75.5 1570 ng/mL 1.3% NA = Not Applicable Samples diluted 1:100 and reported from external standard calibration. Page 24 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS In Groundwater Decatur, AL - December 2011 Table 20. DALGW 134S 111209 PFBS PFHS PFOS 3M LIM S ID D e scrip tio n Concentration (ng/mL) % R ecovery GLP10-01-02-24-031 GLP10-01-02-24-032 GLP10-01-02-24-033 DAL-GW-134S-0-111209 DAL-GW-134S-DB-111209 DAL-GW-134S-FMS-111209 0.361 0.367 18.5 NA NA 89.8 Average Concentration (ng/mL) %RPD 0.364 ng/mL 1.6% Concentration (ng/mL) YoRecovery 4.18 NA 4.25 NA 22.2 89.9 4.22 ng/mL 1.7% C oncentration (ng/mL) YoRecovery 17.5 NA 17.5 NA 34.5 85.0 17.5 ng/mL 0.0% NA = Not Applicable Samples undiluted and reported from external standard calibration. Table 21. DAL GW 134L 111209 PFBS PFHS PFOS 3MLIM SID D e scrip tio n Concentration (ng/mL) % R ecovery GLP10-01-02-24-034 GLP10-01-02-24-035 GLP10-01-02-24-036 DAL-GW-134L-0-111209 DAL-GW-134L-DB-111209 DAL-GW-134L-FMS-111209 10.4 10.6 419 NA NA 81.2 Average Concentration (ng/mL) %RPD 10.5 ng/mL 1.9% C oncentration (ng/mL) YoRecovery 109 NA 109 NA 546 87.6 109 ng/mL 0.0%, C oncentration (ng/mL) YoRecovery 432 NA 439 NA 741 61.2(1) 436 ng/mL 1.6% NA = Not Applicable NO = Not Calculated; Endogenous sample concentration greater than 2x spike level. Samples diluted 1:100 and reported from external standard calibration. (1) FMS did not meet acceptance criteria o f 100 30%. (2) Sample method uncertainty is expanded to 39%. Page 25 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Table 22. DAL GW 135R 111214 PFBS PFHS PFOS 3M LIM S ID D escrip tio n C oncentration (ng/mL) %Recovery GLP10-01-02-24-037 GLP10-01-02-24-038 GLP10-01-02-24-039 DAL-GW-135R-0-111214 DAL-GW-135R-DB-111214 DAL-GW-135R-FMS-111214 63.9 64.9 1820 NA NA 87.3 Average Concentration (ng/mL) %RPD 64.4 ng/mL 1.6% C oncentration (ng/mL) %Recovery 820 833 2740 NA NA 95.7 827 ng/mL 1.6% Concentration (ng/mL) % R ecovery 1130 1080 2580 NA NA 73.8 1110 ng/mL 4.5% NA = Not Applicable Samples diluted 1:100 and reported from external standard calibration. Table 23. DAL GW 135S 111214 PFBS PFHS PFOS 3M LIM S ID D e scrip tio n Concentration (ng/mL) %Recovery GLP10-01-02-24-040 GLP10-01 -02-24-041 GLP10-01-02-24-042 DAL-GW-135S-0-111214 DAL-GW-135S-DB-111214 DAL-GW-135S-FMS-111214 0.961 1.02 19.1 NA NA 89.7 Average Concentration (ng/mL) %RPD 0.991 ng/mL 6.0% C oncentration (ng/mL) YoRecovery 7.78 NA 8.01 NA 26.9 95.0 7.90 ng/mL 2.9%, Concentration (ng/mL) % R ecovery 5.59 NA 6.03 NA 22.8 85.0 5.81 ng/mL 7.6% NA = Not Applicable Samples undiluted and reported from external standard calibration. Page 26 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Table 24. DAL GW 135L 111214 PFBS PFHS PFOS 3MLIM SID D e scrip tio n C oncentration (ng/mL) %Recovery GLP10-01-02-24-043 GLP10-01-02-24-044 GLP10-01-02-24-045 DAL-GW-135L-0-111214 DAL-GW-135L-DB-111214 DAL-GW-135L-FMS-111214 7.06 7.12 200 NA NA 95.5 Average Concentration (ng/mL) %RPD 7.09 ng/mL 0.85% C oncentration (ng/mL) X R e co ve ry 47.2 NA 47.9 NA 232 92.2 47.6 ng/mL 1.5% C oncentration (ng/mL) % R ecovery 32.5 NA 35.1 NA 183 74.6 33.8 ng/mL 7.7% NA = Not Applicable Samples diluted 1:10 and reported from external standard calibration. Table 25. DAL GW 136S 111213 PFBS PFHS PFOS 3M UM SID D escrip tio n Concentration (ng/mL) %Recovery GLP10-01-02-24-049 GLP10-01-02-24-050 GLP10-01-02-24-051 DAL-GW-136S-0-111213 DAL-GW-136S-DB-111213 DAL-GW-136S-FMS-111213 1.80 1.74 106 NA NA 103 Average Concentration (ng/mL) %RPD 1.77 ng/mL 3.4% C oncentration (ng/mL) % R ecovery 11.6 NA 11.2 NA 104 92.8 11.4 ng/mL 3.5% Concentration (ng/mL) % R ecovery 59.9 NA 58.9 NA 145 85.8 59.4 ng/mL 1.7% NA = Not Applicable Samples diluted 1:10 and reported from external standard calibration. Page 27 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS In Groundwater Decatur, AL - December 2011 Table 26. DAL GW 136L 111213 PFBS PFHS PFOS 3M UM SID D escriptio n Concentration (ng/mL) YoRecovery GLP10-01-02-24-052 GLP10-01-02-24-053 GLP10-01-02-24-054 DAL-GW-136L-0-111213 DAL-GW-136L-DB-111213 DAL-GW-136L-FMS-111213 1.88 1.93 104 NA NA 101 Average Concentration (ng/mL) %RPD 1.91 ng/mL 2.6% Concentration (ng/mL) % R ecovery 12.3 NA 12.2 NA 106 93.9 12.3 ng/mL 0.82% Concentration (ng/mL) YoRecovery 65.3 NA 63.4 NA 153 88.8 64.4 ng/mL 3.0%, N A = Not Applicable Samples diluted 1:10 and reported from external standard calibration. Table 27. DAL GW 137S 111209 PFBS PFHS PFOS 3 M LIM S ID D escriptio n Concentration (ng/mL) %Recovery GLP10-01-02-24-058 GLP10-01-02-24-059 GLP10-01-02-24-060 DAL-GW-137S-0-111209 DAL-GW-137S-DB-111209 DAL-GW-137S-FMS-111209 0.127 0.112 1.09 NA NA 96.1 Average Concentration (ng/mL) YoRPD 0.120 ng/mL 13% C oncentration (ng/mL) %Recovery 0.125 0.101 0.949 NA NA 83.8 0.113 ng/mL 21%(1> Concentration (ng/mL) YoRecovery 0.601 0.433 1.32 , NA NA 80.5 0.517 ng/mL 32%<1> N A = Not Applicable Reported results fo r PFBS and PFHS are from external standard calibration. Reported results for PFOS are from internal standard calibration. (1) RPD did not m eet acceptance criteria o f <20%. Page 28 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Table 28. DAL GW 137L 111209 PFBS PFHS PFOS 3MLIM SID D escriptio n Concentration (ng/mL) %Recovery GLP10-01-02-24-061 GLP10-01-02-24-062 GLP10-01-02-24-063 DAL-GW-137L-0-111209 DAL-GW-137L-DB-111209 DAL-GW-137L-FMS-111209 0.0928 0.0807 1.11 NA NA 101 Average Concentration (ng/mL) %RPD 0.0868 ng/mL 14% C oncentration (ng/mL) %Recovery 0.104 0.0943 1.00 NA NA 90.3 0.0992 ng/mL 9.8% Concentration (ng/mL) % R ecovery 1.68 NA 1.55 NA 2.59 97.7 1.62 ng/mL 8.0% NA = Not Applicable Reported results for PFBS and PFHS are from external standard calibration. Reported results for PFOS are from internal standard calibration. Table 29. DAL GW 138R 111214 PFBS PFHS PFOS 3MLIM SID D escriptio n Concentration (ng/mL) %Recovery GLP10-01-02-24-064 GLP10-01-02-24-065 GLP10-01 -02-24-066 DAL-GW-138R-0-111214 DAL-GW-138R-DB-111214 DAL-GW-138R-FMS-111214 7.36 7.43 202 NA NA 96.3 Average Concentration (ng/mL) %RPD 7.40 ng/mL 0.95% C oncentration (ng/mL) %Recovery 35.0 NA 35.5 NA 220 92.4 35.3 ng/mL 1.4% C oncentration (ng/mL) % R ecovery 300 NA 300 NA 451 75.5 300 ng/mL 0.0% NA = Not Applicable Samples diluted 1:10 and reported from external standard calibration. Page 29 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Table 30. DAL GW 138S 111214 PFBS PFHS PFOS 3M UM SID Description C oncentration (ng/mL) X R e cove ry GLP10-01-02-24-067 GLP10-01-02-24-068 GLP10-01-02-24-069 DAL-GW-138S-0-111214 DAL-GW-138S-DB-111214 DAL-GW-138S-FMS-111214 3.71 3.56 191 NA NA 92.8 Average Concentration (ng/mL) %RPD 3.64 ng/mL 4.1% C oncentration (ng/mL) %Recovery 17.9 NA 17.6 NA 203 92.6 17.8 ng/mL 1.7% C oncentration (ng/mL) % R ecovery 159 NA 161 NA 299 69.5 160 ng/mL 1.3% NA = Not Applicable Samples diluted 1:10 and reported from external standard calibration. Table 31. DAL GW 138L 111214 PFBS PFHS PFOS 3MLIM SID Description C oncentration (ng/mL) %Recovery GLP10-01-02-24-070 GLP10-01-02-24-071 GLP10-01-02-24-072 DAL-GW-138L-0-111214 DAL-GW-138L-DB-111214 DAL-GW-138L-FMS-111214 <0.0250 <0.0250 1.00 NA NA 99.0 Average Concentration (ng/mL) %RPD <0.0250 C oncentration (ng/mL) % R ecovery 0.0630 0.0558 0.939 NA NA 88.1 0.0594 ng/mL 12% C oncentration (ng/mL) % R ecovery 0.461 0.379 1.40 NA NA 98.2 0.420 ng/mL 20% NA = Not Applicable Reported results for PFBS and PFFIS are from external standard calibration. Reported results for PFOS are from internal standard calibration. Page 30 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Table 32. DAL GW 140R 111213 PFBS PFHS PFOS 3M UM SID D escrip tio n GLP10-01-02-24-073 DAL-GW-140R-0-111213 GLP10-01-02-24-074 DAL-GW-140R-DB-111213 GLP10-01-02-24-075 DAL-GW-140R-FMS-111213 Average Concentration (ng/mL) %RPD C oncentration (ng/mL) %Recovery 1.23 NA 1.31 NA 97.7 95.5 1.27 ng/mL 6.3% C oncentration (ng/mL) %Recovery 13.9 NA 14.2 NA 109 95.1 14.1 ng/mL 2.1% Concentration (ng/mL) %Recovery 100 NA 103 NA 181 79.7 102 ng/mL 3.0% NA = Not Applicable Samples diluted 1:10 and reported from from external standard calibration. Table 33. DAL GW 141R 111214 PFBS PFHS . PFOS 3M LIM SID D e scrip tio n GLP10-01-02-24-076 DAL-GW-141R-0-111214 GLP10-01-02-24-077 DAL-GW-141R-DB-111214 GLP10-01-02-24-078 DAL-GW-141R-FMS-111214 Average Concentration (ng/mL) %RPD C oncentration (ng/mL) % R ecovery 0.0351 0.0333 1.00 NA NA 95.6 0.0342 ng/mL 5.3% C oncentration (ng/mL) %Recovery 0.259 0.248 1.15 NA NA 89.8 0.254 ng/mL 4.3% C oncentration (ng/mL) % R ecovery 4.97 NA 4.77 NA 5.80 NC 4.87 ng/mL 4.1% m NA = Not Applicable NC = Not Calculated; Endogenous sample concentration greater than 2x spike level. Reported results for PFBS and PFHS are from external standard calibration. Reported results for PFOS are from internal standard calibration. (1) Sampling location did not have an appropriate FMS spike level to sufficiently evaluate PFOS recovery. Page 31 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Table 34. DAL GW 142R 111213 PFBS PFHS PFOS 3MLIM SID D e scriptio n GLP10-01-02-24-079 GLP10-01-02-24-080 GLP10-01-02-24-081 DAL-GW-142R-0-111213 DAL-GW-142R-DB-111213 DAL-GW-142R-FMS-111213 Average Concentration (ng/mL) %RPD C oncentration (ng/mL) % R ecovery 1.83 NA 1.70 NA 48.2 92.0 1.77 ng/mL 7.4% C oncentration (ng/mL) X R e co ve ry 10.2 NA 9.74 NA 53.8 87.8 9.97 ng/mL 4.6% Concentration (ng/mL) %Recovery 94.5 NA 94.2 NA 131 73.4 94.4 ng/mL 0.32% NA = Not Applicable Samples diluted 1:10 and reported from external standard calibration. Table 35. DAL GW 143R 111209 PFBS PFHS PFOS 3MLIM SID D escriptio n C oncentration (ng/mL) %Recovery GLP10-01-02-24-082 GLP10-01-02-24-083 GLP10-01-02-24-084 DAL-GW-143R-0-111209 DAL-GW-143R-DB-111209 DAL-GW-143R-FMS-111209 0.709 0.721 21.6 NA NA 103 Average Concentration (ng/mL) %RPD 0.715 ng/mL 1.7% Concentration (ng/mL) % R ecovery 2.33 NA 2.33 NA 19.6 86.4 2.33 ng/mL 0.0% C oncentration (ng/mL) %>Recovery 16.1 NA 15.1 NA 30.2 73.0 15.6 ng/mL 6.4% NA = Not Applicable Samples diluted 1:10 and reported from external standard calibration. Page 32 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Table 36. DAL GW 144R 111213 PFBS PFHS PFOS 3MLIM SID Description Concentration (ng/mL) %Recovery GLP10-01-02-24-085 GLP10-01-02-24-086 GLP10-01-02-24-087 DAL-GW-144R-0-111213 DAL-GW-144R-DB-111213 DAL-GW-144R-FMS-111213 2.05 1.99 55.9 NA NA 107 Average Concentration (ng/mL) 1 %RPD 2.02 ng/mL 3.0% Concentration (ng/mL) VoRecovery 7.56 NA 7.51 NA 47.4 79.9 7.54 ng/mL 0.66% C oncentration (ng/mL) VoRecovery 26.4 NA 26.9 NA 70.0 86.9 26.7 ng/mL 1.9% NA = Not Applicable Reported results for PFBS and PFHS are from external standard calibration. Reported results for PFOS are from Internal standard calibration. Table 37. Trip Blank 1 3M LIMS ID GLP10-01-02-24-094 GLP10-01-02-24-095 GLP10-01 -02-24-096 GLP10-01-02-24-097 GLP10-01-02-24-098 Description DAL-GW-TRIP01-0 DAL-GW-TRIP01 -LS DAL-GW-TRIP01 -MS DAL-GW-TRIP01 -MHS DAL-GW-TRIP01 -HS PFBS PFHS PFOS Concentration (ng/mL) <0.0250 0.994 19.6 101 499 XoRecovery NA 98.4 97.0 100 99.2 Concentration (ng/mL) <0.0250 0.920 18.5 93.6 467 VoRecovery NA 92.2 92.5 93.8 93.6 Concentration (ng/mL) <0.0371 0.943 19.7 86.5 412 VoRecovery NA 94.5 98.5 86.7 82.6 NA = Not Applicable Reported results from external standard calibration. The MHS sample was diluted 1:10, the HS sample was diluted 1:100. Page 33 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS In Groundwater Decatur, AL - December 2011 Table 38. Rinseate Blanks 3M LIMS ID GLP10-01-02-24-099 GLP10-01-02-24-100 PFBS PFHS PFOS Description DAL-GW-140R-RB-111213 DAL-GW-133L-RB-111214 C oncentration (ng/mL) <0.0250 <0.0250 %Recovery NA NA Concentration (ng/mL) <0.0250 <0.0250 % R ecovery NA NA C oncentration (ng/mL) <0.0371 <0.0371 %Recovery NA NA NA = Not Applicable Reported results from external standard calibration. Table 39. Surrogate Recovery 3M LIMS ID GLP10-01-02-24-058 GLP10-01-02-24-059 GLP10-01-02-24-060 GLP10-01-02-24-061 GLP10-01-02-24-062 GLP10-01-02-24-063 GLP10-01-02-24-070 GLP10-01-02-24-071 GLP10-01-02-24-072 GLP10-01 -02-24-076 GLP10-01-02-24-077 GLP10-01-02-24-078 GLP10-01 -01 -24-085 GLP10-01-01-24-086 GLP10-0.1-01-24-087 Description DAL-GW-137S-0-111209 DAL-GW-137S-DB-111209 DAL-GW-137S-FMS-111209 DAL-GW-137L-0-111209 DAL-GW-137L-DB-111209 DAL-GW-137L-FMS-111209 DAL-GW-138L-0-111214 DAL-GW-138L-DB-111214 DAL-GW-138L-FMS-111214 DAL-GW-141R-0-111214 DAL-GW-141R-DB-111214 DAL-GW-141R-FMS-111214 DAL-GW-144R-0-111213 DAL-GW-144R-DB-111213 DAL-GW-144R-FMS-111213 PFOS surrogate % Recovery 93.5 106 102 101 98.5 91.8 94.0 93.4 97.7 93.7 95.1 97.8 102 96.9 93.9 Page 34 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 10 Conclusion Laboratory control spikes and field matrix spikes were used to determine the analytical method accuracy and precision for PFBS, PFHS, and PFOS. Analysis was successfully completed following 3M Environmental Laboratory method ETS-8-044.1 described herein. 11 Data/Sample Retention All remaining samples and associated project data (hardcopy and electronic) will be archived according to 3M Environmental Laboratory standard operating procedures. Page 35 of 110 12 Signatures GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Page 36 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Attachment A: Protocol A mendment GLP10-01-02; Interim Report 24 Analysis o f PFBS, PFHS, and PFOS In Groundwater Analytical PDerocatotucr,oAl:L G- DLePc1em0-b0e1r -200121 Amendment 24 Study Title Analysis of Perfluorooctane Sulfonate (PFOS), Perfluorohexane Sulfonate (PFHS) and Perfluorobutane sulfonate (PFBS) in Groundwater, Soil and Sediment for the 3M Decatur Phase 3 Site-Related Monitoring Program PROTOCOL AMENDMENT NO. 24 Amendment Date: December 1,2011 Performing Laboratory 3M Environmental, Health, and Safety Operations 3M Environmental Laboratory Building 260-5N-17 Maplewood, MN 55144-1000 Laboratory Project Identification GLP10-01-02 Sampling Event Former Sludge Incorporation Area (FSIA) Groundwater Page 1 of 7 Page 38 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Analytical DPercoattoucr,oAlL: G- DLePce1m0-b0e1r 2-00121 Amendment 24 This amendment modifies the following portion of protocol: "Analysis of PFOS, PFHS and PFBS in Groundwater, Soil and Sediment for the 3M Decatur Phase 3 Site-Related Monitoring Program" Protocol reads: No changes to the wording of the protocol are required. A mend to read: No changes to the wording of the protocol are required. This amendment only addresses and documents the addition of the General Project Outline (GPO) for the collection and analysis of groundwater samples from Decatur, AL, and conducted as part of the 3M Decatur Phase 3 Program for PFOS, PFHS and PFBS (GLP10-01-02). The anticipated sample collection will occur around the timeframe during the week of December 5, 2011. The groundwater samples for this sampling event will be entered into the 3M Environmental Laboratory LIMS as project GLP10-01-02-24 and reported as interim report GLP10-01-0224, (reflecting study GLP10-01-02 and amendment -24). Reason: The reason for this amendment is to document the General Project Outline (GPO) which describes the anticipate groundwater sample collection event to be conducted for the 3M Decatur, AL facility. The GPO is four pages in length and included as attached to this amendment form. Page 2 of 7 Page 39 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Analytical PDercoattoucr,oAl:L G- DLePc1em0-b0e1r 2-00121 Amendment 24 Amendment Approval Page 3 of 7 Page 40 of 110 GLP10-01-02; Interim Report 24 Analysis o f PFBS, PFHS, and PFOS in Groundwater Analytical PDercoattoucr,oAl:L G- DLePc1em0-b0e1r 2-00121 Amendment 24 3 A A Environmental Health & Safety Operations, Environmental Laboratory General Project Outline To: Gary Hohenstein, 3M EHS&Opns From: Susan Wolf, 3M EHS&Opns; Environmental Lab ' ' cc: William Reagen, 3M EHS&Opns; Environmental Lab Cleston Lange, 3M EHS&Opns; Environmental Lab Jai Kesari, Weston Solution's Date: December 1, 2011 Subject: Analysis of Perfluorooctane Sulfonate (PFOS), Perfluorohexane Sulfonate (PFHS) and Perfluorobutane sulfonate (PFBS) in Groundwater, Soil and Sediment for the 3M Decatur Phase 3 Site-Related Monitoring Program; GLP Interim Report 24 - Former Sludge Incorporation Area (FSIA) Groundwater - December 2011 Sampling 1 General Project Information C o n ta c ts Lab Request Number Six D igit Departm ent Number Project Schodule/Test Dates 3M S p o n so r R epresentative Gary Hohenstein 3M EHS Operations 3M Building 224-5W-03 Saint Paul, MN 55144-1000 Phone: (651) 737-3570 aahohenstein@ m m m .com 3M E nvironm ental Laboratory M anagem ent William K. Reagen 3M EHS Opns, Environmental Laboratory 260-5N-17 651 733-9739 wkreaaen@ m m m .com Principal Analytical Investigator Cleston Lange 3M EHS Opns, Environmental Laboratory 260-5N-17 651 733-9860 cclanae@ m m m .com Sam pling Coordinator Timothy Frinak Weston Solutions T im othv.frinak@ w estonsolutions.com Phone: (334)-332-9123 GLP10-01-02-24 Dept #530711, Project #0022674449 Sampling scheduled for the week o f December 5, 2011 All verbal and written correspondence will be directed to Gary Hohenstein. Page 4 of 7 Page 41 of 110 GLP10-01-02; Interim Report 24 Analysis o f PFBS, PFHS, and PFOS in Groundwater Analytical PDercoattoucr,oAl:L G- DLePc1em0-b0e1r 2-00121 Amendment 24 2 Background Information and Project Objective(s) The 3M EHS Operations Laboratory (3M Environmental Lab) will receive and analyze groundwater samples collected from twenty-seven groundwater wells for Perfluorobutanesulfonate (PFBS), Perfluorohexanesulfonate (PFHS), and Perfluorooctanesulfonate (PFOS) from the Former Sludge Incorporation Area (FSIA). Analyses will be conducted under the GLP requirements of EPA TSCA Good Laboratory Practice Standards 40 CFR 792. Groundwater samples from Decatur, AL will be collected by Weston Solutions personnel the week of December 5, 2011. The 3M Environmental Laboratory will prepare the sample bottles with all required spikes to ensure that results for PFBS, PFHS, and PFOS are of a known precision and accuracy. The final report will be submitted to Gary Hohenstein and Jai Kesari upon completion under interim report GLP1001-02-24. 3 Project Schedule Sample collection bottles will be prepared by 3M Environmental Laboratory for sampling the week of December 5, 2011. Sample bottles will be shipped in coolers to 3M Decatur for arrival by Monday, December 5, 2011. Sample bottles should be stored refrigerated on-site until sample collection. Martin Smith \ Weston Trailer 3M Decatur Plant 1400 State Docks Road Decatur, Alabama 35601 4 Test Parameters The targeted limit of quantitation will be 0.025 ng/mL (ppb) for PFBS, PFHS, and PFOS. A total of thirty-one sampling locations have been specified. For each sampling location, a sample, sample duplicate and field matrix spike will be collected. Table 1 indicates the well ID and corresponding field spike level. The "fill to here" line on each 250 mL Nalgene bottle will be 200 mL. One sets of trip blanks consisting of reagent-grade water, a low-level trip blank spike, mid-level field spike, mid-high field spike, and a high-level trip blank spike will be prepared at the 3M Environmental Laboratory and sent to the sampling location with the other bottles. Based on previous sampling of the FSIA (GLP10-01-02-07, GLP10-01-02-011, and GLP10-01-02-18), more than half of the sample locations are expected to have PFHS and/or PFOS levels greater than 100 ng/rriL. To aid in the quantitation of sampling locations with PFBS, PFHS and PFOS levels at expected concentrations of <50 ng/mL, sample bottles will contain the addition of internal standards 'a0 2-PFBS, 13C3-PFHS, and 13C8PFOS, as well as 13C4-PFOS surrogate. The trip blank sample, trip blank low and mid spikes will also include the addition of internal standard standards 180 2-PFBS, 13C3-PFHS, and 13Cs-PFOS, as well as 13C4-PFOS surrogate. Two additional bottles will be prepared to be used for the preparation of the equipment rinseate blanks and will also contain the addition of internal standards 18C>2-PFBS, 13C3-PFHS, and 13CB-PFOS, as well as 13C4-PFOS surrogate. A 1-L bottle of laboratory reagent water will be provided to be used to collect the rinseate blanks. Page 5 of 7 Page 42 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Analytical DPercoattoucr,oAl:L G- DLePc1em0-b0e1r 2-00121 Amendment 24 T able 1. S a m p lin g L o c a tio n s and F ield M atrix S p ike levels. Set# Well No. Spike Cone. (ng/mL) 1 * 136R, 137R, 137S, 137L, 138L, 141R 2 * 131S,134S,135S,143R 1.0 20 3 * 130L, 142R, 144R 50 4 130S, 131L, 133R, 136S, 136L, 140R 5 133S, 133L, 135L, 138R, 138S 100 200 6 130R, 134L, CW26C, CW26L 500 7 131R,134R,135R 2000 Trip Blank ' Sample bottles will contain 80 2-PFBS, surrogate. 1* 20 * 100 500 C3-PFHS, and Ca-PFOS internal standards and 13C4-PFOS 5 Test Methods Samples will be prepared and analyzed by LC/MS/MS following ETS-8-044.1 "Method of Analysis for the Determination of Perfluorinated Compounds In Water by High Performance Liquid Chromatography/Mass Spectrometry Direct Injection Analysis". The data quality objectives for these studies are quantitative results for the target analytes with an analytical accuracy of 10030%. Field matrix spikes not yielding recoveries within 10030% will be addressed in the report and the final accuracy statement may be adjusted accordingly. Based on the results from previously collected samples from the FSIA, most sampling locations will require dilution with laboratory reagent water prior to sample analysis by ETS-8-044.1. 6 Reporting Requirements For each sampling location, the report will contain the results for the sample, sample duplicate, and field matrix spike. Trip blank and trip blank spikes will be reported for the sampling event as will any equipment/rinseate blanks prepared in the field. Laboratory control spikes of reagent water prepared at the time of sample preparation will also be reported and used to evaluate the overall method accuracy and precision. Method blanks of reagent water prepared at the time of sample preparation will be used to determine the method detection limit. For those sampling locations where the field matrix spike level was not appropriate, due to higher than expected analyte concentrations, a laboratory matrix spike may be prepared and included in the final report. 7 Email Correspondence Attachment A: Sampling Bottle Request Page 6 of 7 Page 43 of 110 Attachment A GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Analytical PDerocatotucr,oAl:L G- DLePc1e0m-be1r-20121 Amendment 24 / " f ' \ Bottle request lor 4th quarter 2 0 11 FSIA well sampling Young, Charles T. 11/29/2011 03:10 PM Cc:"Frinak, Timothy R.", "Smith, Martin" From: "Young, Charles T." < Charles.Young@WestonSolutions.com> Sue: Tim Frinak and Martin Smith are setting up for groundwater sampling commencing next week at the following Former Sludge Incorporation Area monitoring wells: 130R 130S 130L 131R 131S 131L 133R 133S 133L 134R 134S 134L 135R 135S 135L 136R 136S 136L 137R 137S 137L 138R 138S 138L 140R 141R 142R 143R 144R CW26C CW26L Can you prepare a bottle set for shipment to Martin Smith's attention at Decatur with arrival by Tuesday, December 6, 2011? As with earlier rounds, analysis will be performed under GLP10-01-01 and GLP1001- 02. Due to the relatively low GW levels at the site, some of the above wells may be dry and therefore will not be sampled. Martin will return any unused bottle sets should some of the locations have insufficient water for sampling. Thanks, Charlie Charles T. Young Technical Director Weston Solutions, Inc. 1400 Weston Way (5-2) West Chester, PA 19380 (610) 701-3787 Fax (610) 701-3401 . CONFIDENTIALITY: This email and attachments may contain information which is confidential and proprietary. Disclosure or use o f any such confidential or proprietary information without the written permission o f Weston Solutions, Inc. is strictly prohibited. If you received this email in error, please notify the sender by return e-mail and delete this email from your system. Thank you. Page 7 of 7 Page 44 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Attachment B: Representative Sample C hromatograms and Calibration Curve(s) Page 45 of 110 Workstation: ETSKIRK GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Results Name: klll227a.rdb Printing Time: 9:46:02 AM ' Printing Date: Wednesday, January 04, 2012 Page 46 of 110 Workstation: ETSKIRK GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS In Groundwater Decatur, AL - December 2011 Results Name: klll227a.rdb k111227a.rdb (PFHS): "Quadratic" Regression ("1 / x" weighting): y = -2.15e+003xA2 + 8.39e+005 x + 4.77e+003 (r = 0.9998) 6.2e7 - 6.0e7- 5.8e7 - 5.6e7 - 5.4e7 - 5.2e7- 5.0e7- 4.8e7- 4.6e7- 4.4e7- 4.2e7- 4.0e7- 3.8e7- 3.6e7- 3.4e7- 3.2e7- 3.0e7- 2.8e7- 2.6e7- 2.4e7 - 2.2e7 - 2.0e7- 1.8e7- 1.6e7- 1.4e7- 1.2e7- 1.0e7- 8.0e6- 6.0e6 - 4.0e6 - 2.0e6- 0.0 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100 Concentration, ng/mL Data worked up by KJU Page 1 of 1 Printing Time: 9:46:32 AM Printing Date: Wednesday, January 04, 2012 Page 47 of 110 Workstation: ETSKIRK GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS In Groundwater Decatur, AL - December 2011 Results Name: klll227a.rdb Data worked up by KJU Printing Time: 9:45:16 AM Printing Date: Wednesday, January 04, 2012 Page 1 of 1 Page 48 of 110 *ETS-Kirk GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Results Name: klll227a.rdb Thursday, January 26, 2012 Page 49 of 110 *ETS-Kirk GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Results Name: klll227a.rdb Data printed by STW Thursday, January 26, 2012 Page 2 of 27 Page 50 of 110 *ETS-Kirk GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Results Name: klll227a.rdb Thursday, January 26, 2012 Page 51 of 110 *ETS-Kirk GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Results Name: klll227a.rdb Data printed by STW Thursday, January 26, 2012 Page 4 of 27 Page 52 of 110 *ETS-Kirk GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS In Groundwater Decatur, AL - December 2011 Results Name: klll227a.rdb Thursday, January 26, 2012 Page 53 of 110 ETS-Kirk Data printed by STW Thursday, January 26, 2012 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Results Name: klll227a.rdb Page 6 of 27 Page 54 of 110 *ETS-Kirk GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Results Name: klll227a.rdb Thursday, January 26, 2012 Page 55 of 110 *ETS-Kirk GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS In Groundwater Decatur, AL - December 2011 Results Name: klll227a.rdb Page 56 of 110 *ETS-Kirk GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Results Name: klll227a.rdb Thursday, January 26, 2012 Page 57 of 110 ETS-Kirk Thursday, January 26, 2012 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Results Name: klll227a.rdb Page 58 of 110 j *ETS-Kirk GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Results Name: klll227a.rdb Data printed by STW Thursday, January 26, 2012 Page 11 of 27 Page 59 of 110 *ETS-Kirk GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Results Name: klll227a.rdb Thursday, January 26, 2012 Page 60 of 110 *ETS-Kirk Data printed by STW Thursday, January 26, 2012 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Results Name: kl!1227a.rdb Page 61 of 110 *ETS-Kirk GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Results Name: klll227a.rdb Data printed by STW Thursday, January 26, 2012 Page 14 of 27 Page 62 of 110 *ETS-Kirk Data printed by STW Thursday, January 26, 2012 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Results Name: kl11227a.rdb Page 63 of 110 *ETS-Kirk GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Results Name: klll227a.rdb Data printed by STW Thursday, January 26, 2012 Page 16 of 27 Page 64 of 110 *ETS-Kirk GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Results Name: klll227a.rdb Data printed by STW Thursday, January 26, 2012 Page 17 of 27 Page 65 of 110 *ETS-Kirk Data printed by STW Thursday, January 26, 2012 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Results Name: klll227a.rdb Page 18 of 27 Page 66 of 110 *ETS-Kirk GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Results Name: kll1227a.rdb Data printed by STW Thursday, January 26, 2012 Page 19 of 27 Page 67 of 110 *ETS-Kirk GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Results Name: klll227a.rdb Page 68 of 110 ETS-Kirk Thursday, January 26, 2012 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Results Name kl!1227a.rdb Page 69 of 110 ETS-Kirk Thursday, January 26, 2012 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS In Groundwater Decatur, AL - December 2011 Results Name klll227a.rdb Page 22 of 27 Page 70 of 110 *ETS-Kirk GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Results Name: kll1227a.rdb Page 71 of 110 ETS-Kirk [Sample Name: *k111227b111* S e m a io 'GLP10-01-02-24-043* File-,k111227b.wir Peak Name *PFOS* Mass(es): *499.000/80.000 Da,499.000/&9.000Da,499.000/130.000 Da* Comment *DAL-GW-135L-0-* Annotabon ~ Sample Type: Unknown Concentration: N/A Acq. Date: Acq Tune- 12/30/2011 9 1:37:48 AM 1Proc. Algorithm: Specify Parameters - MQ 11 Noise Percentage : 50 1 . 0 0 min Peak-Split. Facte Peport Largest Piak: Yes Min. Peak Height:: 0.00 cps Min. Peak width: 0.00 sec Smoothing Width: 3 points RT Window: 30.0 sec Expected RT: 15.4 min Use Relative RT: No 2.6e5 2.4e5 2.2e5 2.0e5 Retention Time: 15.4 min Area. 3262201 counts Height: 3.18e*005 cps End Time: 158 min & 6 0e4 4.0*4 2.0e4 13.0 13.5 I Sample Name: 'k1112276121' Sample ID'*GLP1CM}1-02-24-052* FUa:' k111227b.w4ir Peak Name: `PPOS* Masses): `499.000/80.000 Da.499.000/99.000 Da.499000/130.000 Comment. *DAL-GW-136L-0-` Annotation. " Sample Index: 57 pie Type: Cone N/A Calculated Cone: ng/mL Acq. Date: Acq. Time: 5:04: Modified: Yes Proc. Algorithm: Specify Para Noise Percentage: 50 Base. Sub. Window: 1.00 Peak-Split. Factor: 4 Peport Largest Peak: Yes Min. Peak Height: 0.00 Min. Peak Width: - 0.00 Smoothing Width: 3 RT Window: Expected RT: 15.4 Use Relative RT: Type : Manual >e. 15.5 514006 coi 6.32e005 15.1 15.0 15.5 16.0 16.5 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Results Name: klll227a.rdb ISample Name: >1112276114' Sample ID 'GLP10-01-02-24-049' Fie -kt11227b wtff Peak Name ` PFOS* Mass{e&): '499.000/80.000 Da,499.000/99.000 Da,499.000/130.000 Comment *0AL-GW-136S-0-` Annotation: Sample Index: 50 Sample Type: Unknoi ratior tl/A 1 Cone 59.9 ng/mL 12/30/2011 2-39-44 AM c. Algorithm: Specify Parami se Percentage: 50 e. Sub. Window: 1.00 mi k-Split. Factor: 4 Report Largest Peak: Yes . Peak Height: 0.00 < . Peak Width: 0.00 a Smoothing Width: 3 RT Window: 30.0 sec Expected RT: 15.4 nin Relative RT: No :. Type: Manual ISampte Name: >111227b124' Sampie ID' 'GLP1CKH-02-24-058' Fie. 'k l 11227b.wHT Peak Name -PPOS* Masses): `499.000/80.000 Da,499.000/99.000 Da,499.000/130.000 Da' Comment 'DAL-GW-137S-0-' Annotation: ~ Sample Index: 60 0.587 12/30/201 6:06:15 > ilfied: 1 Proc. Algorithm: Spec se Percentage: k-Split. Factor: Report Large: : Peak: >. Peak Height: >. Peak Width: othlng Width: Window; ected RTUse Relative RT: Int. Type: 14.5 15.0 15.5 16.0 16.5 17.0 17.5 Data printed by STW Thursday, January 26, 2012 15.0 15.5 16.0 16.5 Page 24 of 27 Page 72 of 110 ETS-Kirk Thursday, January 26, 2012 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Results Name: klll227a.rdb Page 25 of 27 Page 73 of 110 ETS-Kirk Data printed by STW Thursday, January 26, 2012 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Results Name: klll227a.rdb Page 26 of 27 Page 74 of 110 ETS-Kirk I Sample Name *k111227t>163* Sample ID: 'GLP10-01-02-24-085* FUe' Peak Name: *PFOS* Masses). '499.000/80.000 03,499.000/99.000 Da,4! Comment "DAL-GW-144R-0-' ArmotaWwr" Sample Index: 99 Noise Percentage- Base. Sub. Window: Peak-Split. Factor: Report Largest Peak: Min. Peak Height: Min. Peak Width: Smoothing Width: RT Window: SC Expected RT: Use s RT: 1.15e5 1.10eS 1.05e5 1.00e5 9.50*4 9.00e4 8.50*4 8.00e4 7.50*4 7.00e4 ^ 6.50e4 6.00e4 ; 5 50*4 = 5,00e4 4.50*4 4.00e4 3.50e4 3.00*4 2.50e4 2.00e4 1.50*4 1.00*4 5000.00 0.00 13.0 13.5 14.0 14,5 15.0 15.5 16.0 16.5 17.0 17.5 Data printed by STW Thursday, January 26, 2012 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Results Name: klll227a.rdb Page 27 of 27 Page 75 of 110 Workstation: ETSBUSTER GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Batch Name: bl20104a.dab Printing Time: 3:41:06 PM Printing Date: Thursday, January 05, 2012 Page 76 of 110 Workstation: ETSBUSTER GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Batch Name: bl20104a.dab Printing Time: 3:41:30 PM Printing Date: Thursday, January 05, 2012 Page 77 of 110 *** Buster J2930203 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Batch Name: bl20104a.dab Data printed by STW Printing Time: 1:29:03 PM Printing Date: Thursday, January 26,' 2012 Page 1 of 8 Page 78 of 110 *** Buster J2930203 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS In Groundwater Decatur, AL - December 2011 Batch Name bl20104a.dab Data printed by STW Printing Time: 1:29:03 PM Printing Date: Thursday. January 26 2012 Page 2 of 8 Page 79 of 110 * Buster J2930203 Data printed by STW Printing Time: 1:29:03 PM Printing Date: Thursday, January 26, 2012 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Batch Name: bl20104a.dab Page 3 of 8 Page 80 of 110 *** Buster J2930203 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Batch Name: bl20104a.dab Printing Time: 1:29:03 PM Printing Date: Thursday, January 26 2012 Page 81 of 110 Buster J2930203 Printing Time: 1:29:03 PM Printing Date: Thursday, January 26, 2012 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Batch Name: bl20104a.dab Page 5 of 8 Page 82 of 110 *** Buster J2930203 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Batch Name: bl20104a.dab Printing Date: Thursday, January 26, 2012 Page 83 of 110 *** Buster J2930203 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS In Groundwater Decatur, AL - December 2011 Batch Name: bl20104a.dab Printing Date: Thursday, January 26, 2012 Page 84 of 110 *** Buster J2930203 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Batch Name: bl20104a.dab Printing Time: 1:29:03 PM Printing Date: Thursday, January 26, 2012 Page 85 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Attachment C: A nalytical Method(s) Page 86 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 3M Environmental Laboratory Method Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis Method Number: ETS-8-044.1 Adoption Date: 4/12/07 Effective Date: I ( \ cf / t / Approved By: William K. Reagen, Technical Director, Environmental Laboratory J / j a is / / Date ETS-8-044.1 Page 1 of 22 Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis Page 87 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 1 Scope and Application This method describes the direct injection analysis of perfluorinated compounds (PFCs) from water matrices using high-performance liquid chromatography tandem mass spectrometry (HPLC/MS/MS). The method is generally applicable but not limited to the measurement of perfluoroalkyl sulfonamides and perfluorinated alkyl acids (PFAAs) such as perfluorosulfonic acids (PFSAs) and perfluorocarboxylic acids (PFCAs) (Table 1). W ater samples containing heavy particulate may require preparation by an alternate method such as ETS-8-154 "Determination of Perfluorinated Acids, Alcohols, Amides, and Sulfonates In W ater By Solid Phase Extraction and High Performance Liquid Chromatography/Mass Spectrometry". The method is applicable to both external standard and internal standard calibration1. Table 1. Representative Target Analytes Acronym PFBA (C4 Acid) PFPeA (C5 Acid) PFHxA (C6 Acid) PFHpA (C7 Acid) PFOA (C8 Acid) PFNA (C9 Acid) PFDA (C10 Acid) PFUnA (C11 Acid) PFDoA (C12 Acid) PFTrDA(C13 Acid) PFBS (C4 Sulfonate) PFHS (C6 Sulfonate) PFOS (C8 Sulfonate) FBSA (C4 Sulfonamide FOSA (C8 Sulfonamide) Analyte Perfluorobutanoic acid Perfluoropentanoic acid Perfluorohexanoic acid Perfluoroheptanoic acid Perfluorooctanoic acid Perfluorononanoic acid Perfluorodecanoic acid Perfluoroundecanoic acid Perfluorododecanoic acid Perfluorotridecanoic acid Perfluorobutanesulfonic acid Perfluorohexanesulfonic acid Perfluorooctanesulfonic acid Perfluorobutanesulfonamide Pefluorooctanesulfonamide Chemical Abstract Services Registry Number (CASRN) 375-22-4 2706-90-3 307-24-4 375-85-9 335-67-1 375-95-1 335-76-2 2058-94-8 307-55-1 72629-94-8 375-73-5 355-46-4 1763-23-1 30334-69-1 754-91-6 The Minimum Reporting Level (MRL) is the Limit of Quantitation (LOQ) that meets Data Quality Objectives (DQOs) that are developed based on the intended use of this method. Method Flexibility - This is a performance-based method and may be generally applied to the determination of perfluorinated compounds in water matrices when analysis batch quality control (QC) criteria are met2. Each set of samples are prepared in an analysis batch with calibration standards, LCSs, blanks, and continuing calibration check standards analyzed on the same instrument during a time period that begins and ends with the analysis of the appropriate continuing calibration check standards. The laboratory is permitted to modify the LC column, mobile phase composition, LC conditions, and MS/MS conditions. Method modifications should be considered to improve method performance or to meet data quality objectives for the study. In all cases where method modifications are implemented, the batch 1The method is supported by validation with internal standard calibration for C4-C13 PFCAs, C4, C6, and C8 PFSAs, and C8 perfluoroalkane sulfonamide in laboratory control samples under 3M method validation El 1-0667. 2Guidance for establishing method QC Criteria based on a.) FDA May 2001, "Guidance for Industry, Bioanalytical Method Validation", b.) EPA Method 537, and c.) European Commission: Guidance for Generating and Reporting Methods o f Analysis in Support of Pre-registration Data Requirements for Annex II (Part A, section 4) and Annex III (Part A,section 5) of Directive 91/414, SANCO/3029/99 rev. 4 (11/07/00). ETS-8-044.1 Page 2 of 22 Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis Page 88 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 analytical QCs (section 9) must be completed and pass QC acceptance criteria (section 13) if the data from the analytical batch are to be reported. 2 Method Summary W ater samples are analyzed as neat aqueous sample or as solvent diluted aqueous samples by direct injection using LC/MS/MS. Samples containing heavy particulate may not be suitable for analysis by this method. Samples containing suspended particulate should be centrifuged or filtered prior to removing a sample aliquot or diluting with solvent. The water sample is mixed well prior to removing an aliquot or diluting, if necessary, with ASTM Type I water, HPLC water, other suitable water, or solvent (methanol). Quantitation is by stable isotope internal standard calibration in laboratory reagent water. All perfluorinated compounds (PFCs) target analyte concentrations of perfluorosulfonic acids (PFSAs) and perfluorocarboxylic acids (PFCAs) are reported as anions and corrected for their salt or free acid forms. Alternatively, quantitation may be performed by external standard calibration. This is a performance-based method. Method uncertainty for each target analyte is determined for each analytical batch using multiple laboratory control spikes at multiple concentrations. This method also requires that the precision and accuracy for each sample be determined using field matrix spikes to verify that the method is applicable to each sample matrix. Calibration standards for PFUnA, PFDoA, PFTrDA, and FOSA have been found to be unstable for more than 2 days in 100% water. Samples requiring analysis for these compounds by this method should be diluted 1:1 with methanol and analyzed against a calibration curve prepared in 1:1 synthetic groundwaterMeOH. 3 Definitions 3.1 Analysis Batch A set of study samples that are prepared with calibration standards, laboratory control samples, and procedural blanks, and analyzed on the same instrument during a time period that begins and ends with the analysis of the appropriate continuing calibration check standards. 3.2 Analytical Sample A portion of a laboratory sample prepared for analysis. 3.3 Calibration Standard A solution prepared by spiking a known volume of the Working Standard (WS) into a predetermined amount of ASTM Type I, HPLC grade water, or other suitable water (i.e. matrix water), and analyzed according to this method. Calibration standards are used to calibrate the instrument response with respect to analyte concentration. 3.4 Laboratory Duplicate Sample (LDS, or Lab Dup) A laboratory duplicate sample is a separate aliquot of a sample taken in the analytical laboratory that is analyzed separately with identical procedures. Analysis of LDSs compared to that of the first aliquot give a measure of the precision associated with laboratory procedures, but not with sample collection, preservation, or storage procedures. ETS-8-044.1 Page 3 of 22 Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis Page 89 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 3.5 Field Blank (FB)/Trip Blank (TB) ASTM Type I, HPLC grade water, or other suitable water, placed in a sample container in the laboratory and treated as a sample in all respects, including exposure to sampling site conditions, storage, preservation and all analytical procedures. The purpose of the TB is to determine if test substances or other interferences are present in the field environment. This sample is also referred to as a Trip Blank. 3.6 Field Duplicate Sample (FDS, Field Dup) A sample collected in duplicate at the same time from the same location as the sample. The FDS is handled under identical circumstances and treated exactly the same throughout field and laboratory procedures. Analysis of the FDS compared to that of the first sample gives a measure of the precision associated with sample collection, preservation and storage, as well as with laboratory procedures. 3.7 Field Matrix Spike (FMS) A sample to which known quantities of the target analytes, ISs and SRSs are added to the sample bottle in the laboratory before the bottles are sent to the field for collection of aqueous samples. A known, specific volume of sample must be added to the sample container without rinsing. This may be accomplished by making a "fill to this level" line on the outside of the sample container. The FMS is analyzed to ascertain if any matrix effects, interferences, or stability issues may complicate the interpretation of the sample analysis. 3.8 Trip Blank Matrix Spike (TBMS) An aliquot of ASTM Type I, HPLC grade water, or other suitable water, to which known quantities of the target analytes, ISs and SRSs are added in the laboratory prior to the shipment of the collection bottles. The TBMS is analyzed exactly like a study sample to help determine if the method is in control and whether a loss of analyte or analytical bias could be attributed to sample holding time, sample storage and/or shipment issues. A low and high TBMS are appropriate when expected sample concentrations are not known or may vary. 3.9 internal Standard (IS) A compound added to each study sample, calibration standard, laboratory control samples, and procedural blanks at a consistent level (typically around 1 ng/mL). The internal standard(s) are stable isotope labeled versions of the target analytes. The area count ratio of the target analyte to the internal standard is used for calibration. Surrogate ISs are applied when stable isotope ISs of target analytes are unavailable. A surrogate IS is not necessarily a stable isotope labeled version of the target analyte, but is treated as an internal standard for quantitation. 3.10 Laboratory Control Sample (LCS) An aliquot of control matrix to which known quantities of the target analytes, ISs and SRSs (when applicable) are added in the laboratory at the time when samples are aliquotted. At least three levels (two levels for SRSs) in triplicate are included, one generally at the low end of the calibration curve and one near the mid range and the upper end of the curve. The LCSs are analyzed exactly like a laboratory sample to determine whether the stability of the standards. LCSs should be prepared each day samples are aliquoted. 3.11 Laboratory Matrix Spike (LMS) A laboratory matrix spike is an aliquot of a sample to which known quantities of target analytes, ISs and SRSs (when applicable) are added in the laboratory. The LMS is analyzed exactly like a laboratory sample to determine whether the sample matrix contributes bias to the analytical results. The endogenous concentrations of the analytes in the sample matrix must be determined in a separate aliquot and the measured values in the LMS corrected for these concentrations. LMSs are optional for analysis of aqueous samples. ETS-8-044.1 Page 4 of 22 Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis Page 90 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 3.12 Laboratory Sample A portion or aliquot of a sample received from the field for testing. 3.13 Limit of Quantitation (LOQ) The lower limit of quantitation (LLOQ) for an analytical batch is the lowest concentration that can be reliably quantitated within the specified limits of precision and accuracy. The LLOQ is generally selected as the lowest non-zero standard in the calibration curve that meets method acceptance criteria. The LLOQ for each target analyte is established for each analysis batch as the lowest calibration standard with area counts at least twice that of the average area counts of the procedural blanks. The upper limit of quantitation (ULOQ) for an analytical batch is the highest concentration that can be reliably quantitated within the specified limits of precision and accuracy. The highest standard in the calibration curve that meets method acceptance criteria is defined as the ULOQ. 3.14 Method/ProceduraD Blank An aliquot of control matrix that is treated exactly like a laboratory sample including exposure to all glassware, equipment, solvents, and reagents that are used with other laboratory samples. The method blank is used to determine if test substances or other interferences are present in the laboratory environment, the reagents, or the apparatus. 3.15 Sample A sample is an aliquot removed from a larger quantity of material intended to represent the original source material. 3.16 Stock Standard Solution (SSS) A concentrated solution of a single-analyte prepared in the laboratory with an assayed reference compound. 3.17 Surrogate Internal Standard An IS that is not necessarily a stable isotopically labeled target analyte, but is treated as an internal standard for quantitation. Surrogate ISs are used when isotopically labeled counterparts of the target analyte are not commercially or readily available. 3.18 Surrogate Recovery Standard (SRS) An isotopically labeled standard, not used as an internal standard, that is added to each sample and appropriate QC sample as a means to evaluate the method performance for a chemical class of compounds (e.g., PFSAs, PFCAs). 3.19 Working Standard (WS) A solution of several analytes prepared in the laboratory from SSSs and diluted as needed to prepare calibration standards and other required analyte solutions. 4 Warnings and Cautions 4.1 Health and Safety The acute and chronic toxicity of the standards for this method have not been precisely determined; however, each should be treated as a potential health hazard. The analyst should wear gloves, a lab coat, and safety glasses to prevent exposure to chemicals that might be present. ETS-8-044.1 Page 5 of 22 Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis Page 91 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 The laboratory is responsible for maintaining a safe work environment and a current awareness of local regulations regarding the handling of the chemicals used in this method. A reference file of material safety data sheets (MSDS) should be available to all personnel involved in these analyses. 4.2 Cautions The analyst must be familiar with the laboratory equipment and potential hazards including, but not limited to, the use of solvents, pressurized gas and solvent lines, high voltage, and vacuum systems. Refer to the appropriate equipment procedure or operator manual for additional information and cautions. 5 Interferences During sample preparation and analysis, major potential contaminant sources are reagents and glassware. All materials used in the analyses shall be demonstrated to be free from interferences under conditions of analysis by running method blanks. Parts and supplies that contain Teflon should be avoided or minimized due to the possibility of interference and/or contamination. These may include, but are not limited to: wash bottles, Teflon lined caps, autovial caps, HPLC parts, etc. The use of disposable micropipettes or pipettes to aliquot standard solutions is recommended to make calibration standards and matrix spikes. 6 Instrumentation, Supplies, and Materials 6.1 Instrumentation Analytical balance capable of reading to 0.0001 g HPLC/MS/MS or HPLC/MS system, as described in Section 10. 6.2 Supplies and Materials Sample collection bottles-- HDPE (e.g., NalgeneTM) wide-mouth bottles with screw cap. Note: Do not use fluorinated or Teflon bottles or lined caps. Coolers or boxes for sample shipment. 15-mL and 50-mL disposable polypropylene centrifuge tubes. Class A pipettes and volumetric flasks, various. 2 mL HPLC autovials Disposable pipettes, polypropylene or glass as appropriate Centrifuge capable of spinning 15-mL and 50-mL polypropylene tubes at 3000 rpm. 7 Reagents and Standards Note: Suppliers and catalog numbers are for illustrative purposes only. Equivalent performance may be achieved using chemicals obtained from other suppliers. Do not use a lesser grade of chemical than those listed. 7.1 Chemicals W ater - Milli-Q, HPLC grade, or other suitably appropriate sources Calcium Acetate - A.C.S. Reagent Grade ETS-8-044.1 Page 6 of 22 Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis Page 92 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Magnesium Acetate - A.C.S. Reagent Grade Methanol - HPLC grade Ammonium Acetate - A.C.S. Reagent Grade 7.2 Representative Target Analytes, ISs, and SRSs PFBA, Heptafluorobutyric Acid, (C4 Perfluorinated Acid) PFPeA, Nonafluoropentanoic Acid (C5 Perfluorinated Acid) PFHxA, Perfluorohexanoic Acid (C6 Perfluorinated Acid) PFHpA, Tridecafluoroheptanoic Acid, (C7 Perfluorinated Acid) PFOA, Ammonium perfluorooctanoate, (C8 Perfluorinated Acid) PFNA, Heptadecafluorononanoic Acid, (C9 Perfluorinated Acid) PFDA, Nonadecafluorodecanoic Acid (C10Perfluorinated Acid) PFUnA, Perfluoroundecanoic Acid, (Cn Perfluorinated Acid) PFDoA, Perfluorododecanoic Acid, (C i2 Perfluorinated Acid) PFTrDA, Perfluorotridecanoic Acid, (C13Perfluorinated Acid) FBSA, Perfluorobutanesulfonamide FOSA, Perfluorooctanesulfonylamide PFBS, Potassium Perfluorobutanesulfonate PFHS, Perfluorohexanesulfonate PFOS, Potassium perfluorooctanesulfonate PFOA [1,2, 3 ,4-13C], 13C4-isotopically labeled perfluorooctanoic acid (SRS) PFOS [1,2, 3 ,4-13C], 13C4-isotopicaily labeled Perfluorooctanesulfonate (SRS) PFUnA [1,2-13C], 13C2-isotopically labeled Perfluoroundecanoic acid (SRS) A custom mix of ISs in a methanolic solution containing ([1,2,3,4-13C4]PFBA, [1,2 13C2]PFHxA, [1,2,3,4,5,6,7,8-13C8]PFOA, [1,2,3,4,5,6,7,8,9-13C9]PFNA, [1,2 -^CJPFDA, [1,2,3,4,5,6,7 -13C7]PFUnA, [1 ,2 -X JP F D o A , [1,2,3 - C3]PFHS, [1,2,3,4,5,6,7,8-13C8]PFOS, and [1,2,3,4,5,6,7,8-13C8]PFOSA (Wellington Laboratories, Guelph, ON) in combination with added ([1,2,3,4,5-13C5]PFPeA, ([1,2,3,4- CJPFHpA, and [^OJPFBS can be used to prepare a stock IS solution. Alternatively, individual stable isotope ISs can be used to prepare a stock IS mixture. Other ISs can be applied. 7.3 Reagent Preparation 2 mM Ammonium acetate solution (Analysis)-- Weigh 0.3 g of Ammonium acetate and dissolve in 2.0 L of reagent water. Synthetic Groundwater (containing 25 ppm Ca and Mg) - Weigh 0.61 g of Calcium Acetate and 0.92 g of Magnesium Acetate and dissolve in 6.0 L of reagent water. Note: Alternative volumes may be prepared as long as the ratios of the solvent to solute ratios are maintained. ETS-8-044.1 Page 7 of 22 Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis Page 93 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 7.4 Stock Standard Solution (SSS) and Working Standard Solution Preparation The following standard preparation procedure serves as an example. Weighed amounts and final volumes may be changed to suit the needs of a particular study. For example, pL volumes may be spiked into volumetric flasks when diluting stock solutions to appropriate levels. 100 pg/mL target analyte SSSs-- Weigh out 10 mg of analytical standard (corrected fo r percent salt, acid [ETS-4-031] and purity) and dilute to 100 mL with methanol or other suitable solvent, in a 100 mL volumetric flask. Transfer to a 125 mL LDPE bottle or other suitable container. Prepare a separate solution for each analyte. Expiration dates and storage conditions of stock solutions should be assigned in accordance with laboratory standard operating procedure. An example of purity and salt correction is given below for PFOS. sa.l.t correct..ion f,act.or = -m---o--le--c--u-l-a--r--w--e--ig- -h--t-o--f--a--n-i-o--nm oclecular weight of salt 499 PFOS (K +)salt correction factor = ------= 0.9275 538 10 mg C8F17S03'K+with purity 90% = 8.35 mg C8F17S03- (10 mg*0.90*0.9275=8.35 mg) 10 pg/mL (10,000 ng/mL) mixed working standard-- Add 5.0 mL each of the 100 pg/mL SSSs to a 50 mL volumetric flask and bring up to volume with solvent. 1 pg/mL (1,000 ng/mL) mixed working standard-- Add 0.5 mL of the 100 pg/mL SSSs to a 50 mL volumetric flask and bring up to volume with solvent. 0.1 pg/mL (100 ng/mL) mixed standard-- Add 0.05 mL of the 100 pg/mL SSSs to a 50 mL volumetric flask and bring up to volume with solvent. Storage Conditions-- Store all SSSs and working standards in accordance with laboratory standard operating procedure or in a refrigerator at 42C for a maximum period of 6 months from the date of preparation. 7.5 Calibration Standards Calibration can be performed by IS or external calibration. Using the working standards described above, prepare calibration solutions in ASTM Type I water, HPLC water, other suitable water, or a mixture of solvent and water using the information in Table 2 as a guideline. Note: Volumes of water or water/solvent mixtures and working standards may be adjusted to meet the data quality objectives addressed in the general project outline. Calibration levels other than those listed below can be prepared as needed. For the quantitation of PFOA and PFOS, reference materials of certified mixed linear and branched isomer are preferred. Alternately, reference materials of primarily linear isomers of PFOA and/or PFOS may be used, however, when quantitating with predominantly linear reference standards, additional LCS samples containing both linear and branched isomers of PFOA and PFOS are required3. 7.5.1 Internal Standard (IS) and Surrogate Recovery Standard (SRS) For IS calibration, stable isotope internal standards of each target analyte or appropriate surrogate ISs should be spiked at the same level in all calibration standards. Once the calibration standards have been prepared as stated above in Section 7.5, all calibration standards are spiked with a separate internal standard spiking solution. Typically the 3A report summarizing an assessment of the use of reference standards containing certified linear and branched isomers of PFOA/PFOS can be found in 3M report Ell-0560. ETS-8-044.1 Page 8 of 22 Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis Page 94 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 concentration of the internal standard is consistent with the internal standard concentration expected in the samples being prepared, usually 1 ng/ml_. The concentration of the internal standard spiking solution is typically 2 pg/mL. A separate zero point or method blank is typically prepared at the same time as the calibration standards, using the same solution used to prepare the standards (ASTM Type I water, HPLC water, other suitable water, or a solvent/water mixture), and is spiked with the internal standard at the same concentration as the calibration curve, typically at 1 ng/mL. If the samples being analzyed were pre-spiked with SRSs, the calibration curve prepared in Section 7.5 is spiked with a separate SRS spiking solution. Typically, the sample bottles are spiked with a SRS at 0.1 ng/mL. The final calibration curve must consist of at least six calibration points after analysis. The following table provides an example of spike concentrations and volumes used to achieve a multi-point extracted calibration curve with internal standard and surrogate standard. Table 1 lists recommended stable isotope internal standards for several PFSA and PFCA target compounds. A custom mix of isotopically labeled target analytes in a methanolic solution containing ([1,2,3,4-13C4]PFBA, [1,2 - CJPFHxA, [1,2,3,4,5,6 7,8-13C8]PFOA, [1,2,3,4,5,6,7,8,9-3C9]PFNA, [1,2,3,4,5,6 -13C6]PFDA, [1,2,3,4,5,6,7 -l i C7]PFUnA, [1,2 X JP F D oA , [1,2,3-13C3]PFHS, [1,2,3,4,5,6,7,8-13C8]PFOS, and [1,2,3,4,5,6,7,8-13C8]FOSA (Wellington Laboratories, Guelph, ON) in combination with added ([1,2,3,4,5-13C5]PFPeA, ([1,2,3,4-13C4]PFHpA, and [1802]PFBS can be used to prepare a stock IS solution. Alternative sources of certified stable isotope labeled target analytes are applicable. Alternatively, individual stable isotope ISs can be used to prepare a stock IS mixture. The table below lists the recommended stable isotope ISs and SRSs applied in the method. Other stable isotope ISs and SRSs of target analytes not listed in the table may be used if supported by validation and/or analysis batch QCs meeting method acceptance criteria (e.g., [13C2]-PFOA). The same internal standard should be used for a given analyte throughout the entire project/study. Note: some of the compounds listed below are appropriate to use as surrogate ISs when a stable isotope IS of a target analyte is not available. Generally, surrogate isotopically labeled PFCAs are used for PFCAs, and surrogate isotopically labeled PFSAs are used for PFSAs. Table 2 provides examples of spike concentrations and volumes used to achieve a multi-point calibration curve with ISs and SRSs. ETS-8-044.1 Page 9 of 22 Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis Page 95 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Table 1. Stable Isotope PFCAs and PFSAs used fo r ISs and SRSs Compound Name Synonym or Acronym 13C4-Perfluorobutanoic acid [1,2,3,4-I3C4]PFBA 13C4-Perfluoropentanoic acid [l,2,3,4,5-l3C5]PFPeA 13C2-Perfluorohexanoic acid [1,2 -i3C2]PFHxA 13C4-Perfluoroheptanoic acid [1,2,3,4-13C4]PFHpA 13C8-Perfluorooctanoic acid [l,2,3,4,5,6,7,8-13C8]PFOA l3C9-Perfluorononanoic acid [1,2,3,4,5,6,7,8,9-'3C9]PFNA 13C6-Perfluorodecanoic acid [1,2,3,4,5,6-i3C6]PFDA 13C7-Perfluoroundecanoic acid [l,2,3,4,5,6,7-l3C7]PFUnA 13C2-Perfluorododecanoic acid [1,2 -,3C2]PFDoA l802-Ammonium Perfluorobutane sulfonate [1s0 2]PFBS l3C3-Ammonium Perfluorohexane sulfonate [1,2,3-13C3]PFHS l3C8-Sodium Perfluorooctane sulfonate [l,2,3,4,5,6,7,8-l3Cg]PFOS 13C8-Perfluorooctanesulfonamide 13C4-Perfluorooctanoic acid [1,2,3,4,5,6,7,8-l3C8]FOSA [1,2,3,4-i3C4]PFOA Analytical Purpose IS for PFBA IS for PFPeA IS for PFHxA IS for PFHpA IS for PFOA and [1,2,3,4 13C4]PFOA IS for PFNA IS for PFDA IS for PFUnA IS for PFDoA, *PFTA IS for PFBS IS for PFHS IS for PFOS and PFOS[1,2,3,4 ,3C4], IS for FOSA SRS for all PFCAs: C4-C8 Reference Standard Source Wellington Labs (Mix or Individual) Wellington Labs . (Mix or Individual) Wellington Labs (Mix or Individual) Wellington Labs (Mix or Individual) Wellington Labs (Mix or Individual) Wellington Labs (Mix or Individual) Wellington Labs (Mix or Individual) Wellington Labs (Mix or Individual) Wellington Labs (Mix or Individual) RTI International (Individual) Wellington Labs (Mix or Individual) Wellington Labs (Mix or Individual) Wellington Labs (mix) RTI International (Individual) Wellington 13C2-Perfluoroundecanoic acid [1,2 -13C2]PFUnA SRS for all PFCAs C9-C13 Wellington l3Cs-Perfluorooctane sulfonate [l,2,3,4-13C4]PFOS SRS for all PFSAs: C4, C6, and C8 Wellington ETS-8-044.1 Page 10 of 22 Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis Page 96 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Table 2. Example Preparation of Calibration Curve with ISs and SRSs Sample Description 0.025 ng/mL curve point 0.030 ng/mL curve point 0.04 ng/mL curve point 0.05 ng/mL curve point 0.1 ng/mL curve point 0.25 ng/mL curve point 0.5 ng/mL curve point 1 ng/mL curve point 2.5 ng/mL curve point 5.0 ng/mL curve point 10.0 ng/mL curve point 25.0 ng/mL curve point 50.0 ng/mL curve point 75.0 ng/mL curve point 100 ng/mL curve point Concentration o f WS, pg/mL 0.10 0.10 0.10 0.10 0.10 0.10 1.0 1.0 10.0 10.0 10.0 10.0 10.0 10.0 10.0 Volume o f WS, pL 25 30 40 50 100 250 50 100 25 50 100 250 500 750 1000 Volume o f IS (2 pg/mL), pL 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 Concentration of Surrogate, pg/mL 0.20 0.20 0.20 0.20 0.20 0.20 0.20 0.20 10.0 10.0 10.0 10.0 10.0 10.0 10.0 Volume of Surrogate, pL 12.5 15 20 25 50 125 250 500 25 50 100 NA NA NA NA Volume o f ASTM Type 1Water, or other suitable so lv e n t<1>, mL 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 N/A - Not Applicable (1) Samples requiring analysis for PFUnA, PFDoA, PFTrDA, and FOSA should be analyzed against a calibration curve prepared in 1:1 synthetic groundwater:MeOFI. ETS-8-044.1 Page 11 of 22 Method of Analysis for the Determination of Perfluorinated Compounds in W ater by LC/MS/MS; Direct Injection Analysis Page 97 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 8 Sample Collection and Bottle Preparation Sample collection bottles are prepared by 3M Environmental Laboratory (or subcontract supplier) personnel for shipment at ambient temperature to the collection site. Typically, four separate collection bottles are associated with a single collection site: sample, field duplicate sample, low field matrix spike, and high field matrix spike. Alternatively, the sample and field duplicate sample may contain SRSs in lieu of additional target analyte low field matrix spike and target analyte high field matrix spike samples. Depending on the scope of the project, additional replicates of the field sample and field matrix spikes may be added. Also, it is not uncommon for additional mid-level field matrix spikes to be collected if the expected sample concentrations are truly unknown or could span a large concentration range. High-density polyethylene (HDPE) wide-mouth Nalgene bottles are used for the sample collection containers. (Volumes of the bottles may vary depending on how much sample is required to meet data quality objectives.) Sample collection volumes are project specific and based on data quality objectives. The Nalgene bottles do not require any pretreatment prior to use. Typically, placement of a sample bottle volumetric "fill to here" line is done by using a sample bottle marker template. Alternatively, bottles may be weighed prior to bottle preparation and weighed again after samples have been collected. All bottles should be clearly labeled to indicate its intended use as a sample, field sample duplicate, low field matrix spike, high field matrix spike, sample/SRS field matrix spike, field duplicate sample/SRS field matrix spike, trip blank, or trip blank matrix spike. If each location has different designated spike levels, the label should also clearly indicate the sample location designation. Generally, a set of bottles for a given collection site are prepared then grouped together in plastic bags for organizational purposes. For each sample collection event, at least one set of trip blank and trip blank matrix spikes are prepared. Bottle preparation should be documented in a Note to File or on a sample preparation worksheet and should include the following information: date prepared, total number of bottles prepared, number of sample sites, the standard identification numbers and spike volumes used to prepare spiked bottles, the "fill to here" volume, and any other pertinent information needed for reconstructibility of the data. The Note to File will be included in the final data package for the project. Samples are collected in the field and shipped to the laboratory at ambient temperature. 8.1 Field Matrix Spike Sample (FMS) Field matrix spike samples are a requirement of the method. A FMS sample is defined as a QC sample to which known quantities of appropriate target analytes are added to the sample bottle in the field or in the laboratory before the bottles are sent to the field. The sample and field duplicate sample may contain appropriate SRSs in lieu of target analyte FMS samples. Sample quantities are determined volumetrically or gravimetrically. A known, specific volume or weight of sample is added to the sample container without rinsing. Volumetric sample measurements may be acquired by a laboratory applied "fill to this level" line on the outside of the sample container. Target analyte FMS samples should be spiked at approximately 0.5-10 times the expected analyte concentration in the sample. If the expected range of analyte concentrations is unknown, multiple spikes at varying levels may be prepared to increase the likelihood that a spike at an appropriate level is made. Typically a low and a high target analyte spike are prepared for each sampling location. In those instances where SRSs are to be used in lieu of target analyte FMS samples, the sample and field duplicate sample are spiked at approximately 2-5 times the target LOQ. The FMS is analyzed to ascertain if matrix effects or sample holding time contributes bias to the analytical results. For the sample bottles designated for matrix spikes, an appropriate volume of matrix spiking solution is added to the empty bottle prior to sampling. The volume of spike solution added should produce the desired final concentration of target analytes once the bottle is filled with sample to the "fill to here line". The matrix spiking solution(s) should be prepared in a suitable solvent and contain all of the appropriate target analytes, ISs, and SRSs. The target analyte matrix spiking solution is often the same as the working standards used to create the calibration standards. An example of a bottle spike is given below. "Fill to here" volume = 200 mL (A 250 mL Nalgene bottle is used) Desired Field Spike Concentration = 0.25 ng/mL ETS-8-044.1 Page 12 of 22 Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis Page 98 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 500 pL of a 0.1 pg/mL spiking solution (containing the target analytes) is added to the bottle and the bottle cap promptly sealed. 8.2 Internal Standard and Surrogate Recovery Standard If analysis of a surrogate recovery standard (SRS) is included in the project objectives, an appropriate volume of a surrogate standard solution is added to all the bottles prior to sampling and SPE. Typically sample bottles are spiked with surrogate recovery standards at a final desired spike concentration of 0.1 ng/mL. If quantitation by internal standard (IS) is included in the project objective, an appropriate volume of internal standard solution is added to all the bottles prior to sampling and SPE. Typically sample bottles are spiked with internal standard at a final desired spike concentration of 1 ng/mL. For the trip blank, the SRS spike and IS spike is added to the bottle and then ASTM Type I water (HPLC grade reagent water or other suitable water may used) is added to the "fill to here" line. The bottle is capped and sealing tape may be placed around the outer edge of the cap. Trip blank matrix spikes are prepared by adding the appropriate volume of target analyte spiking solution, IS, and SRS spiking solutions and filling the bottle to the desired volume with the appropriate water and capping and sealing the cap. 9 Quality Control and Data Quality Objectives 9.1 Data Quality Objectives This method and required quality control samples is designed to generate data accurate to 30% with a targeted LOQ of 0.025 ng/mL. Any deviations from the quality control measures spelled out below will be documented in the raw data and footnoted in the final report. 9.2 Method/Procedural Blanks The method/procedural blank is zero point calibration standard (which includes ISs) analyzed in a regular basis with each analysis batch. At a minimum, method blanks are analyzed prior to instrument calibration, prior to the analysis of CCV samples, after every 10 sample injections, and at the end of the analytical run. The mean area count or area ratios when using internal standard calibration, for each analyte in the method blanks must be less than 50% of the area count counts or area ratios when using internal standard calibration, of the LOQ standard. The standard deviation of the area counts, or area ratios when using internal standard calibration, of these method blanks should be calculated. A specific %RSD acceptance criteria is not specified but is assessed on an analytical batch basis. If the mean area counts or area ratios when using internal standard calibration, of the method blanks exceed 50% of the LOQ standard, then the LOQ must be raised to the first standard level in the curve that meets criteria. Method blanks may be eliminated if technical justification can be provided (e.g. the procedural blank was analyzed after an unexpectedly high level sample). If any procedural blanks are removed from the LOQ determination, document in the raw data and report as appropriate. Laboratory Sample Replicates / Field Duplicate Sample Typically, samples are collected in duplicates in the field. The relative percent difference (RPD) of duplicate samples should be <20% for the precision of sample preparation and analysis to be considered in control. Replicate samples not meeting the <20% RPD criteria are flagged and reported as outside of QC acceptance criteria. 9.3 Laboratory Matrix Spikes (LMSs) LMSs may be performed in lieu of FMSs if FMSs have previously been performed for the sample matrix. Additionally, LMSs may be performed in lieu of FMSs for a sample matrix if the FMS levels were not appropriate for determining spike recoveries relative to endogenous levels of target analytes and appropriate SRSs. Generally, each sample location represents a different sample and sample matrix. LMSs are prepared for each sample and analyzed to determine the matrix effect on spike recovery efficiency of each target analyte and appropriate SRSs. LMSs should be prepared at a minimum of one level and in duplicate. LMS concentrations should be prepared at approximately 0.5-10 times the endogenous concentration or approximately 4-10 times the LOQ concentration of each target analyte. ETS-8-044.1 Page 13 of 22 Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis Page 99 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Lab matrix spike recoveries should fall within 30% of expected values. Sample data with LMS recovery outside of 30% but within 50% of the expected value are flagged and reported as outside of QC acceptance criteria. Data with LMS recovery outside of 50% of the expected value are reported as NR, where NR is defined as "Not Reportable" data outside of QC acceptance criteria. 9.4 Lab Control Sample Lab control spikes are prepared for each analysis batch to determine method accuracy and precision. LCSs should be prepared at three levels in triplicate for each target analyte and at a minimum of two levels in triplicate for appropriate SRSs. Low lab control spikes should be prepared at a concentration in the range of approximately four to ten times higher than the targeted lower LOQ, the mid lab control spikes should be prepared at a concentration near the mid-point of the calibration curve and the high lab control spikes at approximately 80% of the upper LOQ. For each target analyte and SRSs, the percent relative standard deviation (method precision) for each control spike level must be less than or equal to 20% and the average recovery (method accuracy) for each control spike level must be 80-120%. Sample data for target analytes outside of the laboratory control spike acceptance criteria will be handled as follows: If the average recovery of a spiking level falls outside method acceptance, but at least 67% (6 out of 9) of LCS samples are within 20% of their respective nominal value (33% of the QC samples, not all replicates at the same concentration, may be outside 20% of nominal value), the average recovery will be flagged as outside method acceptance criteria. All LCS samples will be control charted as per ETS-4-026. If the average recovery of one of the spiking levels exceeded the analytical method uncertainty as determined by ETS-12012, that analytical batch uncertainty will be expanded for that particular study. If more than 67% of the LCS samples fail to meet method acceptance criteria, the data will not be reported. Calibration standards consisting of mixed branched and linear isomer PFOS/PFOA are preferred. However, for PFOS/PFOA target analytes, if the calibration standards are comprised of predominantly linear isomers only, at least one level of triplicate LCSs should be prepared using PFOS/PFOA which contains a mix of linear and branched isomers. These LCSs will be used to demonstrate quantitative equivalency (or quantitative bias) of the isomeric mix when using a predominantly linear standard for calibration. The mixed linear and branched isomer PFOS/PFOA LCSs recoveries should fall within 30% of expected values. Alternatively, in lieu of mixed branched and linear isomer PFOS/PFOA LCSs, mixed branched and linear isomer PFOS/PFOA TBMSs may be applied to demonstrate method accuracy and precision. 9.5 Field Matrix Spikes (FMSs) / Surrogate Recovery Standards (SRSs) FMSs are prepared for each sampling location and analyzed to determine the matrix effect and sample holding time on the spike recovery of each target analyte and/or appropriate SRS. Generally, each sample location represents a different sample and sample matrix. FMSs are QC samples to which known quantities of appropriate target analytes are added to the sample bottle in the laboratory before the bottles are sent to the field. Typically a low and a high target analyte FMS are prepared for each sampling location. The sample and field duplicate sample may contain appropriate SRSs in lieu of target analyte low field matrix spike and target analyte high field matrix spike samples. Field matrix spike method acceptance criteria are recoveries within 30% of the expected value. If FMS recovery (target analyte or SRS spike) is outside of 30% of the expected value or could not be assessed because the FMS (target analyte) was spiked at an inappropriate level, the sample result is reported as follows: 1. ) If target analyte FMS recovery could not be assessed because the FMS's were at an inappropriate level, then Laboratory Matrix Spikes (LMS) may be substituted. If LMS recoveries are within 30% the data are reportable and flagged to indicate that the FMS spikes levels were inappropriate. 2. ) If multiple target analyte FMS's were prepared on a sample and the closest FMS level to the reported sample meets the 30% acceptance criteria but additional FMS's are outside the 30% acceptance range, the data are reportable and flagged to indicate that while there were failing FMS's, the uncertainty will not be expanded since the most appropriate spike level passed. ETS-8-044.1 Page 14 of 22 Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis Page 100 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 3. ) If the target analyte FMS recoveries are outside of the 30% acceptance range but at least 30 acceptable historical reportable FMS sample results are available, the data may be reported but flagged with an expanded uncertainty and as not meeting FMS criteria. 4. ) Sample data with FMS recovery outside of 30% but within 50% of the expected value are flagged and reported as outside of QC acceptance criteria with an expanded uncertainty. 5. ) If FMS recovery is outside of 50%, the sample result is reported as NR, where NR is defined as "Not Reportable" due to noncompliant QC results. The targeted fortification levels should be at least 50% of the endogenous level and less than 10 times the endogenous level to be used without justification to determine the statement of accuracy for analytical results. Note: It is possible for bottles utilized for Field Matrix Spike samples to be under-filled or over-filled during sample collection. Since this scenario will effect the actual concentration of the FMS sample (surrogate and internal standard concentrations will also be effected, if used), it is important that any obvious under-filling or over-filling of sample bottles be documented in the data package and taken into account in the FMS, ISs, or SRSs recovery calculations. Samples over-filled or under-filled by more than 10% will be require recalculation of the FMS, ISs, and SRS true values. The average of the sample and the field duplicate should be used to calculate the recovery. 10 Procedures 10.1 Water Sample Preparation This method is applicable to water samples. Samples containing heavy particulate may not be suitable for analysis by this method. Samples containing suspended particulate should be centrifuge prior to removing a sample aliquot, or filtered. Thoroughly mix sample before removing an aliquot and placing in a labeled autovial. Dilute sample, if necessary, with ASTM Type I water, HPLC water, other suitable water, or solvent (methanol). Lab control spikes are prepared for each analysis batch to determine method accuracy and precision. LCSs should be prepared at three levels in triplicate for each target analyte and at a minimum of two levels in triplicate for appropriate SRSs. Low lab control spikes should be prepared at a concentration in the range of approximately four to ten times higher than the targeted lower LOQ, the mid lab control spikes should be prepared at a concentration near the mid-point of the calibration curve and the high lab control spikes at approximately 80% of the upper LOQ. For IS quantitation, stable isotope internal standards of each target analyte or appropriate surrogate ISs should be spiked at the same level as the samples being analyzed, in all LCSs. If LCSs are being prepared using synthetic groundwater, allow the LCSs samples to equilibrate for a minimum of 4 hours before aliquoting for analysis or diluting with solvent (methanol). 11 Sample Analysis - LC/MS/MS 11.1 Instrument Setup Note: In this example, an Applied Biosystems Sciex API 4000 (API 5000 or API 5500) Tandem Mass Spectrometer (LC/MS/MS) is used. Other brands/models of LC/MS/MS instruments as well as single quadrupole mass spectrometers (LC/MS) may be used as long as the method acceptance criteria are met. Brand names, suppliers, part numbers, and models are for illustrative purposes only. Equivalent performance may be achieved using apparatus and materials other than those specified here, but demonstration of equivalent performance that meets the requirements of this method is the responsibility of the laboratory. The operator must optimize and document the equipment and settings used. ETS-8-044.1 Page 15 of 22 Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis Page 101 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Establish the LC/MS/MS system and operating conditions equivalent to the following: Mass Spec: Applied Biosystems API 4000, API 5000, or API 5500 Ion Source: Turbo Ion Spray (ABS) Mode: Electrospray Negative Scan Type: MRM (Multiple Reaction Monitoring) Computer: Dell DHM Software: Windows 2000 or Windows XP, Analyst 1.4.2 or higher versions HPLC: Agilent Series 1100,1200, or 1290 Agilent Quaternary Pump Agilent Vacuum Degasser Agilent Autosampler Agilent Column Oven Note: One or more C18 HPLC analytical columns (2.1 mm x 100 mm, 5pm or 2.1 mm x 50 mm, 5pm) may be attached on-line after the purge valve and before the sample injection port to retard and separate any residue contaminants that may be in the mobile phase and/or HPLC system. HPLC Column: Betasil C18,4.6mm x 100mm, 5pm (ThermoElectron Corporation) Column Temperature: 35C Injection Volume: 5pL Mobile Phase (A): 2mM Ammonium Acetate in ASTM Type I water (See 7.3) Mobile Phase (B): Methanol Table 3. Liq u id C hrom atography G radient Program . Step Number 0 1 2 3 4 5 Total Time (min) 0 2.0 14.5 15.5 16.5 20.0 Flow Rate ( u U m in ) 750 750 750 750 750 750 Percent A (2 mM ammonium acetate) 97.0 97.0 5.0 5.0 97.0 97.0 Percent B (Methanol) 3.0 3.0 95.0 95.0 3.0 3.0 Note: Other HPLC gradients may be used as long as the method criteria and project data quality objectives are met. It may be necessary to adjust the HPLC gradient in order to optimize instrument performance. Columns with different dimensions (e.g. 2.1mm x 30mm) and columns from different manufacturers (Keystone Betasil C18 etc.) may be used. ETS-8-044.1 Page 16 of 22 Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis Page 102 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS In Groundwater Decatur, AL - December 2011 Table 4 Suggested MRM Transitions for Target Analytes, Surrogates, and Internal Standards Analyte PFBA (C4 Acid) PFPeA (C5 Acid) PFHxA (C6 Acid) PFHpA (C7 Acid) PFOA (C8 Acid) PFNA (C9 Acid) PFDA (CIO Acid) PFUnA (Cl 1Acid) PFDoA(C12 Acid) PFTA (C l3 Acid) FBSA (C4 Sulfonamide) FOSA (C8 Sulfonamide) PFBS (C4 Sulfonate) PFHS (C6 Sulfonate) PFOS (C8 Sulfonate) il,2,3,4 -I3C41PFBA il,2,3,4,5-'3C5lPFPeA il,2 -13C21PFHxA n,2,3,4- 13C4lPFHpA ri,2,3,4,5,6,7,8-13C8lPFOA il ,2,3,4,5,6,7,8,9-13C9lPFNA ri,2,3,4,5,6-13C61PFDA ri.2,3,4,5,6,7-13C,lPFUnA n,2 -13C21PFDoA i180 2lPFBS n,2,3-'3C3lPFHS il,2,3,4- 13C4]PFOS il,2,3,4,5,6,7,8-13C8lFOSA ri,2,3,4-13C4lPFOA H,2,3,4- 13c 4ipfos n,2 -13C2lPFUnA Analyte Description Target Target Target Target Target Target Target Target Target . . Target Target Target Target Target Target IS for PFBA IS for PFPeA IS for PFHxA IS for PFHpA IS for PFOA IS for PFNA IS for PFDA IS for PFUnA IS for PFDoA and PFTA IS for PFBS IS for PFHS IS for PFOS IS for FOSA Surrogate (C4-C8 Acids) Surrogate)Sulfonates, FOSA) Surrogate (C9-C13 Acids) Mass Transition 01 (amu) 213 263 313 363 413 463 513 563 613 663 298 498 299 399 499 217 268 315 367 421 472 519 570 615 303 402 503 507 417 503 565 M ass Transition Q3 (amu) 169 219 269, 119 319, 169 369,219,169 419, 169,219 469,269,219 519,269,219 569, 169,319 619, 369,319 78 78 99, 80 99, 80 80,99, 130 172 223 270 322 376 427 474 525 570 84 80 80 80 372 80 520 Multiple transitions for monitoring the analytes is an option. The use of one daughter ion is acceptable if data sensitivity and selectivity is achieved and provided that retention time criteria are met to assure adequate specificity. While the daughter ions may be chosen at the discretion of the analyst, mass transition 99 is suggested for PFOS. Quantitation may be performed using the total ion chromatogram (TIC, or summed MRMs) for a given analyte. For example, the PFOA TIC would sum all three of the monitored transitions. Use of the suggested primary ion is recommended. Retention times may vary slightly, on a day-to-day basis, depending on the batch of mobile phase and the gradient, column, guard column(s) used etc. Drift in retention times is acceptable within an analytical run, as long as the drift continues through the entire analysis and the standards are interspersed throughout the analytical run. 11.2 Calibration Curve Quantitation is by internal standard or external standard calibration. Calibration standards may be prepared in ASTM Type I, HPLC water, other suitable water, or a solvent/water mixture. If internal standard calibration does not meet calibration acceptance criteria, external calibration can be applied. See Table 1 for ETS-8-044.1 Page 17 of 22 Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis Page 103 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 recommended application of available internal standards. Quantitation of PFOA and PFOS is by summed analyte-specific mass transitions. Analyze the standard curve prior to each set of samples. If internal standards were added to the calibration standards area ratios are used to generate the calibration curve. The standard curve may be plotted using a linear regression (y = mx + b), weighted 1/x or unweighted, or by quadratic fit (y = ax2 + bx + c), weighted 1/x or unweighted, using suitable software. The mathematical method used to calculate the calibration curve should be applied consistently throughout a study. Any change should be thoroughly documented in the raw data. High and/or low points may be excluded from the calibration curves to provide a better fit over the range appropriate to the data or because they did not meet the pre-determined acceptance criteria. Low-level curve points should also be excluded if their area counts (or area ratio if quantitating by IS) are not at least twice that of the average area counts (or area ratio if quantitating by IS) of method and/or solvent blanks. The coefficient of determination (r2) value for the calibration curve must be greater than or equal to 0.990 (or a correlation coefficient (r) of 0.995). Each point in the curve must be within 25% of the theoretical concentration with the exception of the LLOQ, which may be within 30%. Justification for exclusion of calibration curve points will be noted in the raw data. A minimum of 6 points will be used to construct the calibration curve. If the calibration curve does not meet acceptance criteria, perform routine maintenance or prepare a new standard curve (if necessary) and reanalyze. 11.3 Continuing Calibration Verification (CCV) Continuing calibration verifications (CCV) are analyzed to verify the accuracy of the calibration curve. Analyze a mid-range calibration standard, one of the same standards used to construct the calibration curve, at a minimum after every tenth sample, not including solvent blanks, with a minimum of one per sample set. Calibration verification injections must be within 25% to be considered acceptable. The calibration curve and the last passing CCV will then bracket acceptable samples. Multiple CCV levels may be used. Samples must be bracketed by passing CCVs or the calibration curve and a passing CCV to be reportable. 11.4 System Suitability A minimum of three system suitability samples should be injected at the beginning of each analytical run, prior to the analysis of the calibration curve. Typically these samples are at a concentration near the mid-level of the calibration curve and are repeated injections from one autosampler vial. It is suggested that the system suitability injections have area counts or area ratios when using internal standard calibration, with a target RSD of 5% and a target retention time RSD of <2%. There is no defined acceptability limit on these results as the %RSD value is dependent on the number of MRM transitions being monitored in the LC/MS/MS run or time period. Ultimately, any effects on these parameters for the System Suitability samples will also be evident on all standards and QC samples analyzed aspart of the analysis batch. Any effect of system suitability is incorporated within QC acceptance criteria.4 11.5 Sample Analysis and QCs For each analysis batch, the instrument analysis run sequence should include an initial calibration curve, samples, FDSs, interspersed blanks, interspersed CCVs, appropriate QCs (i.e., LCSs, LMSs, FMSs, TBMSs, and TBs), and a final CCV or calibration curve bracketing samples and appropriate QCs Inject the same volume (between 5 -1 00pL) of each standard, analytical sample and blank into the instrument (unless an on-instrument sample dilution is desired). Samples containing analytes that are quantitated above the concentration of the highest standard in the curve should be further diluted and reanalyzed. 4 3M Environmental Laboratory study E08-0096 evaluated the effect on these results as a function of the number of MRMs being monitored. ETS-8-044.1 Page 18 of 22 Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection ' Analysis Page 104 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 12 Data Analysis and Calculations The chromatography analysis software will typically calculate the amount of target analyte in the sample extracts using the established calibration curve. Calculate the percent recovery of the LCS using the following equation: LCS Concentration ( -^ - ) LCS% recovery = ---------------------------- m L _ * j qo% ng Spike Concentration (----) mL Calculate the percent recovery of the LMS using the following equation: LMS % recovery LMS Concentration (-25.) - Concentration of Sample (-25.) _________________ mL________________________mL Spike Concentration (-25.) mL 100% For samples fortified with known amounts of analyte prior to extraction, use the following equation to calculate the percent recovery. Recovery = Total analyte found (ng/mL) - Average analyte found in sample (ng/mL) X 100 Analyte added (ng/mL) 13 Analysis Batch Method Performance Criteria Any method performance parameters that are not achieved must be considered in the evaluation of the data. Nonconformance to any specified parameters must be described and discussed in the final report if the Technical Manager (non-GLP study) or Study Director (GLP study) chooses to report the data. If criteria listed in this method performance section are not met, maintenance may be performed on the system and samples reanalyzed, or other actions taken as appropriate. Document all actions in the raw data. If data are to be reported when performance criteria have not been met, the data must be footnoted on tables and discussed in the text of the report. 13.1 System Suitability - Analysis Batch A minimum of three system suitability samples should be injected at the beginning of each analytical run. These samples are run prior to the calibration curve. It is suggested that the system suitability injections have area counts with a target RSD of <5% and a target retention time RSD of <2%. There is no defined acceptability limit on these results as the %RSDs are dependent on the number of MRM transitions being monitored in the LC/MS/MS run or time period. Any effect of system suitability is incorporated in the QC acceptance criteria. 13.2 Calibration and Limit of Quantitation (LOQ) - Analysis Batch C alibration Curve: The coefficient of determination (r2) value for the calibration curve must be greater than or equal to 0.990 corresponding to a correlation coefficient (r) = 0.995. Each point in the curve must be within +25% of the theoretical concentration with the exception of the LLOQ, which may be within 30%. CCV Perform ance: The calibration standards that are interspersed throughout the analytical sequence are evaluated as continuing calibration verifications in addition to being part of the calibration curve. The accuracy ETS-8-044.1 Page 19 of 22 Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis Page 105 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS In Groundwater Decatur, AL - December 2011 of each curve point must be within 25% of the theoretical value (within 30% for lowest curve point). Samples that are bracketed by CCVs not meeting these criteria must be reanalyzed. Lim its o f Q uantitation (LOQ): The lower LOQ (LLOQ) is the lowest non-zero active standard in the calibration curve; the peak area of the LLOQ must be at least 2X that of the average area counts for all prepared procedural blank(s). By definition, the measured value of the LLOQ must be within 30% of the theoretical value. Dem onstration o f S pecificity: Specificity is demonstrated by chromatographic retention time (within 4% of standard) and the mass spectral response of unique ions. 13.3 Blanks - Method/Procedural Blanks and Trip M ethod/Procedural Blanks: Multiple procedural blanks should be interspersed throughout the analysis batch and the analytical sequence. At a minimum, method blanks are analyzed prior to instrument calibration, prior to the analysis of CCV samples, after every 10 sample injections, and at the end of the analytical run. The mean area counts (or area ratios when using IS calibration) for each analyte must be less than 50% of the area count of the LOQ standard. If the area counts of the procedural blanks exceed 50% of the LOQ standard, then the LOQ must be raised to the first standard level that meets criteria. Trip Blank: A trip blank of ASTM Type I water (or lab equivalent) is prepared in a sample container in the laboratory and treated as a sample, including exposure to shipping, sampling site conditions, storage, preservation and all analytical procedures. The trip blanks results for each analyte are included with the reported sample results. 13.4 Data Accuracy and Precision - Analysis Batch Lab C ontrol Spikes: The average recovery at each LCS level for each target analyte and appropriate SRS should be within 80-120% and the percent relative standard deviation of the recoveries must be less than or equal to 20%. If the average recovery of a spiking level falls outside method acceptance, but at least 67% (6 out of 9) of LCS samples are within 20% of their respective nominal value (33% of the QC samples, not all replicates at the same concentration, may be outside 20% of nominal value), the average recovery will be flagged as outside method acceptance criteria. All LCS samples will be control charted as per ETS-12-012. If the average recovery of one of the spiking levels exceeded the analytical method uncertainty as determined by ETS-12-012, that analytical batch uncertainty will be expanded for that particular study. The average recovery at each LCS level for mixed branched/linear isomer PFOA and PFOS should be within 70-130% and the percent relative standard deviation of the recoveries must be less than or equal to 20%. Field D uplicates: The relative percent difference (RPD) of duplicate samples should be less than 20% for the precision of sample preparation and analysis to be considered in control. Replicate samples not meeting the 20% RPD criteria are flagged and reported as outside of QC acceptance criteria. Field M atrix Spikes: FMS acceptance criteria are recoveries within 30% of the expected value for each target analyte and appropriate SRS. Sample data with FMS recovery outside of 30% but within 50% of the expected value are flagged and reported as outside of QC acceptance criteria. Data with FMS recovery outside of 50% of the expected value are reported as NR, where NR is defined as "Not Reportable" data outside of QC acceptance criteria. If FMS recovery could not be assessed because FMSs were at an inappropriate level, then Laboratory Matrix Spikes (LMSs) may be substituted. If LMS recoveries are within 30% for each target analyte and SRSs the data are reportable but flagged as not meeting the FMS method acceptance criteria. 13.5 Analytical Method Uncertainty Analytical method uncertainty for each target analyte and SRS is determined with control charted historical analysis batch LCS data for the method and reported with each analysis batch.5 Uncertainty determinations 5 Method uncertainty based on INTERNATIONAL ANS/ISO/IED STANDARD 17025 reference (GUM, Guide to the Expression of Uncertainty in Measurement). Method application demonstrated in ETS-12-012, citing references: a.) EURACHEM/CITAC Guide, "Quantifying Uncertainty in Analytical Measurement," Second Edition; Editors: S.L.R. Ellison, M. Rosslein, and A. Williams. b.)Georgian, Thomas, "Estimation of Laboratory Analytical Uncertainty Using Laboratory Control Samples," Environmental Testing & ETS-8-044.1 Page 20 of 22 Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis Page 106 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS In Groundwater Decatur, AL - December 2011 are based on INTERNATIONAL ANS/ISO/IED STANDARD 17025 reference (GUM, Guide to the Expression of Uncertainty in Measurement) and described in ETS-12-012. At least thirty data points are required for determining analytical method uncertainty. The method uncertainty is defined as 2x the standard deviation of the percent recoveries of the pooled lab control spikes. While all LCS data points are control charted, only the most recent fifty data points are used for determining the method uncertainty. When less than thirty LCS data points have been generated for a given analyte, the analysis batch LCSs are used to determine the data uncertainty. If FMSs meet the 30% recovery criteria at a level appropriate to the endogenous level, and the LCS meet the 20% recovery criteria, then the uncertainty of the data is determined as within 10020%. Analysis batch sample data with FMS recovery outside of 30% but within 50% of the expected value are flagged and reported as outside of QC acceptance criteria with expanded uncertainties. Data with FMS recovery outside of 50% of the expected value are reported as NR, where NR is defined as "Not Reportable" data outside of QC acceptance criteria. If FMS recovery could not be assessed because FMSs were at an inappropriate level, then Laboratory Matrix Spikes (LMSs) may be substituted. If LMS recoveries are within 30% for each target analyte and appropriate SRSs the data are reportable but flagged as not meeting the FMS method acceptance criteria with uncertainties of 30%. If FMS do not meet the 30% recovery criteria, and historical FMS data does not exist, the analytical uncertainty is evaluated on a sample-by-sample basis, the data may be reported with expanded uncertainty and are flagged. 13.6 Quantitation of PFOA/PFOS - Analysis Batch Calibration standards consisting of mixed branched and linear isomer PFOS/PFOA are preferred. Quantitation is performed by integrating the linear and branched isomers together. Alternately, the linear and branched isomers can be integrated separately, applying the appropriate true value to each calibration curve point for each isomer. The LCS and samples are then quantitated by integrating the linear and branched isomers separately (requires separate analytical results files) and quantitating the resulting peak against the linear or branched calibration curve. The results from both integrations are then summed to produce the final result. Integrating the linear and branched isomers separately may be helpful for those samples where the linear/branched ratios do not closely match those of the reference standards. However, for PFOS/PFOA target analytes, if the calibration standards are comprised of predominantly linear isomers only the method requires the addition of LCSs of mixed branched/linear isomer PFOS/PFOA. The purpose of including these LCSs is to demonstrate quantitative equivalency (or quantitative bias) of the isomeric mix when using a predominantly linear PFOS or PFOA standard for calibration. Alternatively, in lieu of mixed branched and linear isomer PFOS/PFOA LCSs, mixed branched and linear isomer PFOS/PFOA TBMSs may be applied to demonstrate method accuracy and precision. An alternate method of quantitation can be performed whereby only the linear isomer of PFOS/PFOA is integrated and used for generating the calibration curve. The LCS and samples are then quantitated by integrating the linear and branched isomers separately (requires separate analytical results files) and quantitating the resulting peak against the linear calibration curve. The results from both integrations are then summed to produce the final result. Integrating the linear and branched isomers separately reduces the oncolumn concentration for those samples that contain both linear and branched isomers of PFOA/PFOS. This ensures that the concentration detected is within the a range of the calibration curve that is comparable regardless of whether the calibration curve was generated using predominantly linear isomers of PFOS/PFOA or linear plus branched isomers of PFOS/PFOA. 14 Pollution Prevention and Waste Management Waste generated when performing this method will be disposed of appropriately. The original samples will be archived at the 3M Environmental Laboratory in accordance with internal procedures. Analysis, November/December 2000. c.)Taylor, B.N. and CE. Kuyatt, NIST Technical Note 1297, 1994 Edition: "Guidelines for Evaluating and Expressing the Uncertainty of NIST Measurement Results."d.)Adams, T.M., "A2LA Guide for the Estimation of Measurement Uncertainty in Testing", July 2002. ETS-8-044.1 Page 21 of 22 Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis Page 107 of 110 GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 15 Records Each data package generated for a study must include all supporting information for reconstruction of the data. Information for the data package must include, but is not limited to the following items: study or project number, sample and standard prep sheets/records, instrument run log (instrument batch records, instrument acquisition method, summary pages), instrument results files, chromatograms, calibration curves, and data calculations. 16 Affected Documents None. 17 Revisions Revision Number 1 Summary of Changes Section 1. Included the use of internal standard calibration by this method. Section 2. Included the use of internal standard calibration by this method. Included the use of a solvent/water mixture when analyzing for PFUnA, PFDoA, PFTrDA, and FOSA. Section 3. Added definitions for internal standard, surrogate internal standard, and surrogate recovery standard. Section 6.Removed the details regarding the instrument parameters to section 10 of the method. Section 7. Updated reference standards to include internal standards and surrogates. Changed concentration levels for working standards and included the use of internal standards and surrogates. Section 8. Inserted a new section on sample bottle preparation. Section 9 Quality Control. This section was previously section 10 in ETS-8-044.0. Updated QC criteria to be consistent with method ETS-8-154.4. Section 10 Procedures. This section was previously section 8 (Sample Handling) in ETS-8044.0. Added detail regarding the preparation of LCSs. Included the use of methanol as a dilution solvent. Section 11 Sample Analysis. This section was previously section 10 in ETS-8-044.0. Included the details regarding the instrument parameters. Section 12 Data Analysis and Calculations. This section was previously section 11 in ETS8-044.0. Removed the equation for calculating the analytes concentration, indicating that this is done by the instrument software. Section 13 Method Performance. This section was previously section 12 in ETS-8-044.0. Updated QC criteria to be consistent with ETS-8-154.4. Added information on the determination of analytical method uncertainty and quantitation of PFOA/PFOS. Section 14 Pollution Prevention. This section was previously section 13 in ETS-8-044.0. Section 15 Records. This section was previously section 14 in ETS-8-044.0. Section 16 Affected Documents. This section was previously section 15 in ETS-8-044.0. Section 17 Revisions. This section was previously section 16 in ETS-8-044.0. ETS-8-044.1 Page 22 of 22 Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis Page 108 of 110 Attachment D: Deviation(s) GLP10-01-02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 Page 109 of 110 GLP10-01 -02; Interim Report 24 Analysis of PFBS, PFHS, and PFOS in Groundwater Decatur, AL - December 2011 R ecord of D eviation/N onconform ance 1. Identification Study / Project No. G LP 10 -0 1-02-24 Date(s) of Occurrence: 12/27/11, 1/4/12 Document Number: ETS-8-044.1 Deviation type (Check one) SOP Equipment Procedure 0 Method Protocol GPO Other: II. Description (attach extra pages as needed) Method Requirements: 1. FMS recovery within 30% (section 13.4). 2. RPD values <20% (section 13.4). Actual procedure/process: 1. The sampling location DAL GW 134L had a PFOS FMS recovery of 61.2%. 2. The sampling location DAL GW 137S had a sample/sample duplicate RPD of 21% for PFHS and 32% for PFOS. (such as amIIeIn.dmAecnttiiossnuesd,TSaOkPernevision, etc.) Corrective Action ( Yes 0 No) Reference: Acceptability of the nonconforming work: 1. The sampling location with a FMS recovery outside method acceptance criteria will be flagged in the report and the analytical uncertainty for DAL GW 134L will be expanded to 39% for PFOS. 2. The non-compliant RPDs will be flagged in the final report. Actions: Halting of W ork Client Notification W ork Recall W ithholding of Report 0 Other: Deviations will be noted in final report. Project Lead/PAI Approval: Date:y y Study Director (if GLP): Sponsor Approval (for GLP protocol deviations): NA Technical Reviewer (optional): NA Date Date: Date: NA Laboratory Department Manager Approval: Date: 3 ) J^o)o/X ____ Where halting of work occuIrVre.d, Areusutmhpotiroinzaoftiwoonrk mtousRt feirsstubemaeppWrovoerdkby Laboratory Management Laboratory Department Manager Approval: NA Date: NA Deviation No. (assigned by Study Director or Team Leader at the end of study or project) ETS-4-008.7 Page 1 of 1 Documentation of Deviations and Control of Nonconforming Testing Pase 110 of 110