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GLP10-01-02; Interim Report 38 - Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Collected at the Former Sludge Incorporation Area (FSIA) located in Decatur, AL, November/December 2012 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 Weston Way West Chester, PA 19380 P hone: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 GLP10-01-02-38 Total Number of Pages 109 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 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL This page has been reserved for specific country requirements. Page 2 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS In Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL GLP Compliance Statement Report Title: GLP10-01-02; Interim Report 38 - Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Collected at the Former Sludge Incorporation Area (FSIA) in Decatur, AL, November/December 2012 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 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Quality Assurance Statement Report Title: GLP10-01-02; Interim Report 38 - Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Collected at the Former Sludge Incorporation Area (FSIA) in Decatur, AL, November/December 2012 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 Phase 01/08/13 Data / Report m=r QAU Represepgtive Date Reported to Testing Facility Management Study Director 1/11/13 1/11/13 0//V/3 Date Page 4 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Table of Contents GLP Compliance Statement.......................................................................................................................3 Quality Assurance Statement.....................................................................................................................4 Table of Contents........................................................................................................................................5 List of T a b le s...............................................................................................................................................6 1 Study Information.................................................................................................................................8 2 Sum m ary..............................................................................................................................................9 3 Introduction......................................................................................................................................... 11 4 Test & Control Substances...............................................................................................................11 5 Reference Substances......................................................................................................................12 6 Test S ystem ....................................................................................................................................... 13 7 Method S um m ary..............................................................................................................................13 7.1 M ethods.............................................................................................................................13 7.2 Sample Collection..............................................................................................................13 7.3 Sample Preparation...........................................................................................................13 7.4 Analysis..............................................................................................................................14 8 Analytical Results...............................................................................................................................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 8.8 Field Matrix Spikes (FMS).................................................................................................18 Page 5 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 9 Data Summary and Discussion........................................................................................................19 10 Conclusion..........................................................................................................................................32 11 Data/Sample Retention..................................................................................................................... 32 12 Attachm ents.......................................................................................................................................32 13 Signatures..........................................................................................................................................33 List of Tables Table 1. Summarized PFBS, PFHS, and PFOS Results (FSIA GW, Nov/Dec 2012).........................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...........................................................................................................18 Table 10. DAL GW 130R 121130.......................................................................................................... 20 Table 11. DAL GW 130S 121130........................................................................................................... 20 Table 12. DAL GW 130L 121130........................................................................................................... 21 Table 13. DAL GW 131R 121205.......................................................................................................... 21 Table 14. DAL GW 131S 121204........................................................................................................... 22 Table 15. DAL GW 131L 121205........................................................................................................... 22 Table 16. DAL GW 133R 121205.......................................................................................................... 23 Table 17. DAL GW 133S 121205........................................................................................................... 23 Table 18. DAL GW 133L 121205........................................................................................................... 23 Table 19. DAL GW 134R 121205.......................................................................................................... 24 Table 20. DAL GW 134S 121205........................................................................................................... 24 Page 6 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Table 21. DAL GW 134L 121205........................................................................................................... 24 Table 22. DAL GW 135R 121205.......................................................................................................... 25 Table 23. DAL GW 135S 121205........................................................................................................... 25 Table 24. DAL GW 135L 121205........................................................................................................... 25 Table 25. DAL GW 136S 121130........................................................................................................... 26 Table 26. DAL GW 136L 121130........................................................................................................... 26 Table 27. DAL GW 137S 121205........................................................................................................... 27 Table 28. DAL GW 137L 121205........................................................................................................... 27 Table 29. DAL GW 138S 121205........................................................................................................... 27 Table 30. DAL GW 138L 121205........................................................................................................... 28 Table 31. DAL GW CW26C 121205......................................................................................................28 Table 32. DAL GW CW26L 121205........................................................................................................28 Table 33. DAL GW 140R 121130...........................................................................................................29 Table 34. DAL GW 141R 121130...........................................................................................................29 Table 35. DAL GW 142R 121130...........................................................................................................30 Table 36. DAL GW 145R 121204...........................................................................................................30 Table 37. Trip B lank................................................................................................................................ 31 Table 38. Rinseate Blanks..................................................................................................................... 31 Attachments Attachment A: Protocol Amendment 38 (General Project Outline) Attachment B:Representative Chromatograms and Calibration Curve Attachment C: Analytical Method - ES-8-044.1 Attachment D: Protocol Amendment Deviation 34 41 85 108 Page 7 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 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@westonsolutions.com Study Location Testing Facility 3M EHS Operations 3M Environmental Laboratory Building 260-5N-17 St. Paul, MN 55144 Study Personnel William K. Reagen, Ph.D., 3M Laboratory Manager Cleston Lange, Ph.D., Principal Analytical Investigator, (clange@mmm.com) : phone (651)-733-9860 Susan Wolf, 3M Analyst Chelsie Grochow, Analyst Kelly Ukes, Analyst Study Dates Study Initiation: March 8, 2010 Interim 38 Experimental Termination: January 5, 2013 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 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 2 Summary The 3M Environmental Laboratory received groundwater samples from wells located at the Former Sludge Incorporation Area (FSIA) in Decatur, AL, for perfluorobutane sulfonate (PFBS), perfluorohexane sulfonate (PFHS), and perfluorooctane sulfonate (PFOS) analysis under 3M project GLP10-01 -02-38. A total of seventy-five sample bottles were prepared for thirty-one different sampling locations. Due to dry condition, samples were not collected from four of the specified sampling locations; DAL GW 136R, DAL GW 137R, DAL GW 138R, and DAL GW 143R. Samples were collected November 30 - December 5, 2012 and received from Weston personnel on December 7, 2012. The 3M Environmental Laboratory prepared sample containers for thirty-one sampling locations. For each sampling location, a minimum of two sample bottles were prepared (primary sample and a field sample duplicate). The 3M Environmental Laboratory has demonstrated in previous sampling at the FSIA, that Environmental Laboratory Method ETS-8-044.1 is appropriate for the given sample matrix, based on acceptable recoveries of target analyte field matrix spikes (FMS). Therefore, for this sampling event, a field matrix spike sample was prepared for six of the twenty-seven sampling locations collected. In addition, a trip blank set containing Milli-QTM water and appropriate trip blank spikes were prepared, as well as two equipment rinseate blanks. The equipment rinseate blanks did not have FMS samples prepared for determination of PFBS, PFHS, or PFOS recovery. All of the samples were prepared and analyzed for PFBS, PFHS, and PFOS following 3M Environmental Laboratory Method ETS-8-044.1. 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 68.7 ng/mL. The PFHS concentration results for all groundwater samples ranged from 0.0571 ng/mL to 720 ng/mL. The PFOS concentration results for all groundwater samples ranged from 0.380 ng/mL to 1430 ng/mL. The analytical method uncertainties associated with the reported results using external calibration are: PFBS 24% and PFHS 17%. Samples analyzed using internal calibration for PFOS have an analytical method uncertainty of 36%. Page 9 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Table 1. Summarized PFBS, PFHS, and PFOS Results (FSIA GW, Nov/Dec 2012). Sampling Location DAL GW 130R 121130 DAL GW 130S 121130 DAL GW 130L 121130 DAL GW 131R 121205 DAL GW 131S 121204 DAL GW 131L 121205 DAL GW 133R 121205 DAL GW 133S 121205 DAL GW 133L 121205 DAL GW 134R 121205 DAL GW 134S 121205 DAL GW 134L 121205 DAL GW 135R 121205 DAL GW 135S 121205 DAL GW 135L 121205 DAL GW 136S 121130 DAL GW 136L 121130 DAL GW 137S 121205 DAL GW 137L 121205 DAL GW 138S 121205 DAL GW 138L 121205 DAL GW CW26C 121205 DAL GW CW26L 121205 DAL GW 140R 121130 DAL GW 141R 121130 DAL GW 142R 121130 DAL GW 145R 121204 TriD Blank ('Milli-Q Water) GLP10-01-02-38 Equipment rinseate blanks: DAL GW 131S and DAL GW 145R PFBS (1) Avg. Conc. (ng/mL) RPD 14.2 1.4% 3.09 3.6% 1.37 9.5% 36.7 7.9% 0.821 1.3% 2.68 4.5% 3.01 2.0% 11.3 5.3% 8.87 0.90% 38.4 2.9% 0.409 8.1% 8.88 1.0% 68.7 3.6% 1.05 5.7% 7.04 4.7% 2.53 6.3% 2.24 17% 0.294 1.4% 0.0764 7.9% 4.81 2.5% <0.0250 4.72 4.2% 11.8 7.7% 1.45 2.1% 0.0516 19% 1.63 12% 1.59 4.4% <0.0250 <0.0250 PFHS (1) Avg. Conc. (ng/mL) RPD 109 5.5% 27.4 5.1% 10.1 7.2% 210 2.4% 4.68 5.1% 18.2 8.3% 20.5 2.9% 74.6 0.13% 66.5 4.8% 358 1.7% 4.63 11% 90.7 1.9% 720 2.4% 10.5 2.9% 49.7 4.0% 18.8 3.7% 15.0 4.0% 0.203 0.49% 0.0918 8.9% 22.4 2.2% 0.0571 7.0% 50.1 0.80% 95.5 0.52% 15.5 3.9% 0.339 0.59% 9.30 3.2% 7.43 2.3% <0.0250 <0.0250 PFOS (2) Avg. Conc. (ng/mL) RPD 541 19% 144 6.3% 70.9 22% (3) 1430 0.70% 7.48 5.1% 39.7 0.50% 44.2 0.23% 198 4.6% 443 5.4% 943 8.3% 21.4 9.3% 484 1.2% 1100 0.0% 8.06 9.2% 46.0 5.2% 115 6.1% (4) 92.2 5.8% 1.01 6.8% 1.41 2.1% 231 2.2% 0.380 5.3% 384 0.78% 910 4.4% 150 11% 6.08 5.6% (5) 105 9.7% 118 3.4% <0.0278 <0.0278 (1) Sample results reported using external standard calibration. The analytical method uncertainties associated with the reported results using external calibration are: PFBS 24% and PFHS 17%. (2) Sam ple results reported using internal standard calibration. The analytical method uncertainty associated with the reported results using internal calibration is 36% for PFOS. (3) The RPD did not meet method acceptance criteria of <20%. (4) The FMS recovery did not meet acceptance criteria. The analytical uncertainty has been adjusted for PFOS to 42%. See section 9 o f the report for additional information. (5) The sample FMS was not appropriate to assess recovery for PFOS. Page 10 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 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 bottle sets for each groundwater sampling location included a field sample and field sample duplicate. A field matrix spike samples was prepared for six of the twenty-seven sampling locations collected. Each empty container for groundwater samplings 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 and branched) prior to being sent to the field for sample collection. See section 8.8 of the report for field matrix spike levels. 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 Water by High Performance Liquid Chromatography/Mass Spectrometry Direct Injection Analysis". All samples were analyzed for PFOS with the addition of internal standard 13C8-PFOS to used to aid in the data quality objectives. 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. 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. Page 11 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 5 Reference Substances R eferen ce S ubstance Chemical Name Chemical Formula Identifier Use Source Expiration Date Storage Conditions Chemical Lot Number TCR Number Physical Description Purity PFBS (p red o m in an tly lin e a r isom er) Perfluorobutane sulfonate C 4F gS O 3- K + NA Target Analyte Reference Standard 3M 1/10/2017 Frozen 101 TCR-121 W hite Powder 96.7% PFHS (lin e a r isom er) Perfluorohexane sulfonate C aF 13S O 3 Na L-PFHXS Target Analyte Reference Standard W elling to n 3/25/2015 Frozen LPFHxSAM08 TCR12-0021 Crystalline >98% PFOS (b ran ch ed and lin e a r isom ers) Perfluorooctane sulfonate C 8F 17S O 3- K+ Br-PFOSK Target Analyte Reference Standard W elling to n 12/1/2014 Frozen Br-PFOSK1111 TCR12-0026 Liquid 100% R eferen ce S ubstance Chemical Name Chemical Formula Identifier Use Source Expiration Date Storage Conditions Chemical Lot Number TCR Number Physical Description Purity PFOS (b ran ch ed and lin e a r isom ers) Perfluorooctane sulfonate C 8 F 17S O 3 - K+ CAS # 2795-39-3 FMS Reference Standard Aldrich 2/4/2014 Room Temperature 1424328V TCR11-0028 W hite Powder 99.7% 13c 8-p f o s Sodium Perfluorooctanesulfonate 13C8F17SO3-N a + MPFC-C-0112 Internal Standard W elling to n 1/24/2015 Frozen 012312 TCR12-0004, TCR120036 Liquid NA (1) 13c 8-p f o s Sodium Perfluorooctanesulfonate 13C8F17SO3-Na+ MPFC-C-1012 Internal Standard W elling to n 10/3/2015 Frozen 100212 TCR12-0038 Liquid NA (1) (1) Custom mixture of seven mass-labeled (13C) perfluoroalkylcarboxylic acids, two mass labeled (13C) perfluoroalkylsulfonates and one mass-labeled (13C) perfluoro-1-octanesulfonamide at a nominal concentration of 5 pg/mL Page 12 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 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 0 121205 Sampling Location Well ID Sampling Point Well Level Sampling Date Sample Type Example DAL = Decatur, Alabama GW = Groundwater Example: 131 R = Residuum shallow water-bearing zone L = Bedrock water-bearing zone S = Epikarst middle water-bearing zone 121205 - December 5, 2012 0=primary sample DB=duplicate sample FMS = Held 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 Water 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 GLP10-01-02-38 were returned to the laboratory at ambient conditions on December 7, 2012. 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. A number of the sampling locations required dilution. Samples were prepared by diluting 1 mL sample with 9 mLs of Milli-QTM water (dilution 1:10). Samples requiring a 1:100 dilution were prepared by diluting 0.1 mL of sample with 9.9 mLs of Milli-QTM water Samples were analyzed for PFOS using internal standard calibraton. The samples were prepared by adding an aliquot of an internal standard spiking solution to an aliquot of the sample or diluted sample (nominal concentration of 1 ng/mL). Laboratory control samples were prepared in a similar manner. Page 13 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 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 in Table 3, Table 4 and Table 5. 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 ETS Buster ETS-8-044.1 12/26/12 - PFBS and PFHS External standard calibration Agilent 1100 Betasil C18 (4.6 mm X 100 mm), 5u Betasil C18 (4.6 mm X 100 mm), 5u 10 uL Applied Biosystems API 4000 Turbo Spray Turbo ion electrode Negative Analyst 1.6.1 ETS Kirk ETS-8-044.1 1/3/13 - PFOS Internal standard calibration Agilent 1260 Betasil C18 (4.6 mm X 100 mm), 5u Betasil C18 (4.6 mm X 100 mm), 5u 10 uL Applied Biosystems Triple Quad 5500 Turbo Spray Turbo ion electrode Negative Analyst 1.6.1 Table 4. Liquid Chromatography Conditions. Step Number 0 1 2 3 4 5 6 7 8 9 Total Time (min) 0.0 0.5 4.0 6.0 11.0 13.0 13.5 16.0 16.5 19.0 Flow Rate (fL/min) Percent A (2 m M ammonium acetate) ETS-8-044.1 Analysis 750 90.0 750 90.0 750 70.0 750 70.0 750 20.0 750 20.0 750 10.0 750 10.0 750 90.0 750 90.0 Percent B (Methanol) 10.0 10.0 30.0 30.0 80.0 80.0 90.0 90.0 10.0 10.0 Page 14 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Table 5. Mass Transitions. Analyte Mass Transition Q1/Q3 Reference Material Structure Internal Standard Mass Transition Q1/Q3 PFBS 299/80 299/99 Linear NA NA PFHS 399/80 399/99 Linear NA NA 499/80 PFOS 499/99 Branched and Linear [ 13Cs]P F O S (1) 507/80 499/130 Dwell tim e was 75 m sec for each transition. The individual transitions were sum m ed to produce a "total ion chrom atogram " (TIC), which was used for quantitation. (1) Internal standard was only used for the analysis of PFOS. 8 Analytical Results 8.1 Calibration 8.1.1 External standard calibration Samples were analyzed for PFBS and PFHS 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-QTM 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. 8.1.2 Internal standard calibration Samples were analyzed for PFOS 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 reagent water. A separate internal standard spiking solution was prepared and an aliquot was added at the same level to all calibration standards at a nominal concentration of 1 ng/mL. A calibration curve ranging from 0.025 ng/mL to 100 ng/mL (nominal) was analyzed. A quadratic, 1/x weighted, calibration curve of the peak area ratio was used to fit the data for PFOS. 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. For both external and internal standard calibration, 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 or peak area ratio and retention time criteria of less than or equal to 2% RSD was met for PFBS, PFHS, and PFOS. Page 15 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 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. Limit of Quantitation (LOQ). Analysis Date 12/26/12 Analysis external standard calibration 1/3/13 Analysis internal standard calibration NA = Not Applicable Dilution 1 10 100 1 10 100 PFBS LOQ, ng/mL 0.0250 0.250 2.50 NA NA NA PFHS LOQ, ng/mL 0.0250 0.250 2.50 NA NA NA PFOS LOQ, ng/mL NA NA NA 0.0278 0.278 2.78 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, and PFOS. 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, and PFOS. 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 analyzed with the sample dilutions by external standard calibration were prepared by spiking known amounts of PFBS and PFHS into Milli-QTM water to produce the desired concentration. LCSs analyzed by internal standard calibration were prepared by spiking known amounts of PFOS into laboratory reagen water and adding an aliquot of a separate internal standard spiking solution at a nominal concentration of 1 ng/mL. The method acceptance criteria, average of LCS at each level should be within 100% 20% with an RSD <20%, were met for all LCS samples with the exception of the low level LCSs for PFOS, which had an overall average recovery of 127%. A method deviation is included with the raw data. All LCSs were used in the determination of the analytical method uncertainty in section 8.7. The following calculations were used to generate data in Table 7 for laboratory control spikes. LCS Percent Recovery -C--a--l-c-u--l-a--t-e-d---C--o--n--c-e--n--t-r-a--t-io--n- **1. 00% Spike Concentration LCS% RSD = standard deviation LCS replicates * 1QQ% average LCS recovery Page 16 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Table 7. Laboratory Control Spike Recovery. ETS-8-044.1 Analyzed 12/26/12 External standard calibration Lab ID PFBS Spiked Concentration (n g /m L ) Calculated Concentration (n g /m L ) %Recovery Spiked Concentration (n g /m L ) PFHS Calculated Concentration (n g /m L ) LCS-121220-1 LCS-121220-2 LCS-121220-3 Average %RSD 0.498 0.498 0.498 0.482 0.494 0.451 95.5 4.7% 96.8 99.2 90.5 0.498 0.498 0.498 0.524 0.514 0.508 103% 1.5% LCS-121220-4 LCS-121220-5 LCS-121220-6 Average %RSD 4.98 4.98 4.98 4.77 4.88 4.80 96.7% 1.3% 95.7 98.1 96.4 4.98 4.98 4.98 5.14 5.17 5.09 103% 0.97% LCS-121220-7 LCS-121220-8 LCS-121220-9 Average %RSD 29.9 29.9 29.9 26.7 25.4 25.5 86.5% 2.7% 89.2 85.0 85.4 29.9 29.9 29.9 28.5 26.5 27.5 92.0% 3.6% %Recovery 105 103 102 103 104 102 95.2 88.6 92.1 ETS-8-044.1 Analyzed 1/3/13 Internal standard calibration Lab ID PFOS Linear and Branched) Spiked Calculated Concentration Concentration (n g /m L ) (n g /m L ) %Recovery LCS-130102-1 LCS-130102-2 LCS-130102-3 Average %RSD 0.184 0.184 0.184 0.233 0.233 0.232 127% 0.46% (1) 127 127 126 LCS-130102-4 LCS-130102-5 LCS-130102-6 Average %RSD 1.84 1.84 1.84 2.24 2.20 2.19 120% 1.3% 122 120 119 LCS-130102-7 LCS-130102-8 LCS-130102-9 Average %RSD 18.4 18.4 18.4 20.0 20.0 19.8 109% 0.53% 109 109 108 (1) LCS average recovery did not meet method acceptance criteria of 100 20%. Page 17 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 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 Calibration External External Internal Standard Deviation 11.9 8.48 17.8 Method Uncertainty 24% 17% 36% 8.8 Field Matrix Spikes (FMS) A field matrix spike (FMS) was collected at six sampling locations 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 and 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 137S and 141R 131S 130S, 136S, and 145R Trip Blank Spike Level FMS FMS FMS Low Mid High PFBS, ng/mL 2.00 19.9 99.6 2.00 19.9 99.6 PFHS, ng/mL 2.00 19.7 98.6 2.00 19.7 98.6 PFOS, ng/mL 1.85 20.0 100 1.85 20.0 100 FMS R (Sample Concentration of FMS - Average Concentration :Field Sample & Field Sample Dup.) Spike Concentraton Page 18 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 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 136S - The recovery of the FMS sample for PFOS was 58.5%. Based on the recovery of the FMS sample, the analytical uncertainty has been expanded for PFOS to 42%. Due to the limited number of FMS samples prepared, the overall average FMS recovery was calculated to determine whether the analytical uncertainty should be expanded for all sampling locations. The overall average FMS recovery was determined to be 84.9%. Since the overall average FMS recovery was greater than 70%, meeting the acceptance criteria of 30%, the method uncertainty will only be expanded for sampling location DAL GW 136S. DAL GW 130L - The sample/sample duplicate RPD was 22% for PFOS. DAL GW 141R - The sample FMS was not appropriate to assess recovery for PFOS. Page 19 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Table 10. DAL GW 130R 121130 3M LIMS ID Description GLP10-01 -02-38-001 GLP10-01 -02-38-002 DAL-GW -130R-0-121130 D AL-G W -130R -D B -121130 Average Concentration (ng/mL) %RPD Samples diluted 1:100. Table 11. DAL GW 130S 121130 PFBS Concentration (n g /m L ) 14.3 14.1 14.2 ng/m L 1.4% PFHS Concentration (n g /m L ) 106 112 109 ng/m L 5.5% PFOS Concentration (n g /m L ) 489 593 541 ng/mL 19% 3M LIMS ID Description GLP10-01 -02-38-003 DAL-GW -130S-0-121130 GLP10-01 -02-38-004 DAL-GW -130S-DB-121130 GLP10-01 -02-38-005 DAL-GW -130S-FMS-121130 Average Concentration (ng/mL) %RPD NA = Not Applicable Samples diluted 1:10. PFBS PFHS PFOS Concentration (n g /m L ) %Recovery 3.14 NA 3.03 NA 91.4 88.7 3.09 ng/m L 3.6% Concentration (n g /m L ) %Recovery 28.1 NA 26.7 NA 108 81.7 27.4 ng/m L 5.1% Concentration (n g /m L ) %Recovery 148 NA 139 NA 228 84.5 144 ng/m L 6.3% Page 20 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Table 12. DAL GW 130L 121130 3M LIMS ID Description GLP10-01 -02-38-006 DAL-GW-130L-0-121130 GLP10-01 -02-38-007 DAL-GW -130L-DB-121130 Average Concentration (ng/mL) %RPD PFBS Concentration (n g /m L ) 1.30 1.43 1.37 ng/m L 9.5% NA = Not Applicable Samples diluted 1:10. (1) Sample/sample duplicate RPD did not meet acceptance criteria of <20%. Table 13. DAL GW 131R 121205 PFHS Concentration (n g /m L ) 9.77 10.5 10.1 ng/m L 7.2% PFOS Concentration (n g /m L ) 63.1 78.6 70.9 ng/m L 2 2 % (1) 3M LIMS ID Description GLP10-01 -02-38-008 DAL-GW-131R-0-121205 GLP10-01 -02-38-009 DAL-GW -131R-DB-121205 Average Concentration (ng/mL) %RPD Samples diluted 1:100. PFBS Concentration (n g /m L ) 38.1 35.2 3 6 .7 ng/m L 7.9% PFHS Concentration (n g /m L ) 207 212 210 ng/m L 2.4% PFOS Concentration (n g /m L ) 1420 1430 1430 ng/m L 0.70% Page 21 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Table 14. DAL GW 131S 121204 3M LIMS ID Description GLP10-01 -02-38-010 GLP10-01 -02-38-011 GLP10-01 -02-38-012 D A L-G W -131S -0-121204 D AL-G W -131S-D B-121204 DAL-GW-131S-F MS-121204 Average Concentration (ng/mL) %RPD Sample GLP10-01 -02-38-012 diluted 1:10. PFBS PFHS PFOS Concentration (n g /m L ) %Recovery 0.815 0.826 17.4 NA NA 83.2 0.821 ng/m L 1.3% Concentration (n g /m L ) %Recovery 4.56 4.80 19.9 NA NA 77.2 4.68 ng/m L 5.1% Concentration (n g /m L ) %Recovery 7.29 7.67 22.9 NA NA 77.1 7.48 ng/m L 5.1% Table 15. DAL GW 131L 121205 3M LIMS ID Description GLP10-01 -02-38-013 DAL-GW-131L-0-121205 GLP10-01 -02-38-014 DAL-GW-131L-DB-121205 Average Concentration (ng/mL) %RPD Samples diluted 1:10. PFBS Concentration (n g /m L ) 2.74 2.62 2.68 ng/m L 4.5% PFHS Concentration (n g /m L ) 18.9 17.4 18.2 ng/m L 8.3% PFOS Concentration (n g /m L ) 39.8 39.6 39.7 ng/m L 0.50% Page 22 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Table 16. DAL GW 133R 121205 3M LIMS ID Description GLP10-01 -02-38-015 GLP10-01 -02-38-016 D A L-G W -133R -0-121205 D A L-G W -133R -D B -121205 Average Concentration (ng/mL) %RPD Samples diluted 1:10. Table 17. DAL GW 133S 121205 PFBS Concentration (n g /m L ) 2.98 3.04 3.01 ng/m L 2.0% PFHS Concentration (n g /m L ) 20.2 20.8 20.5 ng/m L 2.9% PFOS Concentration (n g /m L ) 44.1 44.2 44.2 ng/m L 0.23% 3M LIMS ID Description GLP10-01 -02-38-017 DAL-GW-133S-0-121205 GLP10-01 -02-38-018 DAL-GW-133S-DB-121205 Average Concentration (ng/mL) %RPD Samples diluted 1:10. Table 18. DAL GW 133L 121205 PFBS Concentration (n g /m L ) 11.6 11.0 11.3 ng/m L 5.3% PFHS Concentration (n g /m L ) 74.5 74.6 74.6 ng/m L 0.13% PFOS Concentration (n g /m L ) 193 202 198 ng/mL 4.6% 3M LIMS ID Description GLP10-01 -02-38-019 DAL-GW-133L-0-121205 GLP10-01 -02-38-020 DAL-GW-133L-DB-121205 Average Concentration (ng/mL) %RPD Samples diluted 1:10. PFBS Concentration (n g /m L ) 8.91 8.83 8.87 ng/m L 0.90% PFHS Concentration (n g /m L ) 64.9 68.1 66.5 ng/m L 4.8% PFOS Concentration (n g /m L ) 455 431 443 ng/m L 5.4% Page 23 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Table 19. DAL GW 134R 121205 3M LIMS ID Description GLP10-01 -02-38-021 DAL-GW -134R-0-121205 GLP10-01-02-38-022 DAL-GW-134R-DB-121205 Average Concentration (ng/mL) %RPD Samples diluted 1:100. Table 20. DAL GW 134S 121205 PFBS Concentration (n g /m L ) 37.8 38.9 38.4 ng/m L 2.9% PFHS Concentration (n g /m L ) 361 355 358 ng/m L 1.7% PFOS Concentration (n g /m L ) 904 982 943 ng/m L 8.3% 3M LIMS ID Description G LP 10-01-02-38-023 DAL-GW -134S-0-121205 GLP10-01-02-38-024 DAL-GW-134S-DB-121205 Average Concentration (ng/mL) %RPD Table 21. DAL GW 134L 121205 PFBS Concentration (n g /m L ) 0.425 0.392 0.409 ng/m L 8.1% PFHS Concentration (n g /m L ) 4.89 4.36 4.63 ng/m L 11% PFOS Concentration (n g /m L ) 22.4 20.4 21.4 ng/m L 9.3% 3M LIMS ID Description GLP10-01 -02-38-025 DAL-GW-134L-0-121205 GLP10-01-02-38-026 DAL-GW-134L-DB-121205 Average Concentration (ng/mL) %RPD Samples diluted 1:100. PFBS Concentration (n g /m L ) 8.92 8.83 8.88 ng/m L 1.0% PFHS Concentration (n g /m L ) 89.8 91.5 90.7 ng/m L 1.9% PFOS Concentration (n g /m L ) 481 487 484 ng/m L 1.2% Page 24 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Table 22. DAL GW 135R 121205 3M LIMS ID Description GLP10-01 -02-38-027 GLP10-01 -02-38-028 D A L-G W -135R -0-121205 D A L-G W -135R -D B -121205 Average Concentration (ng/mL) %RPD Samples diluted 1:100. Table 23. DAL GW 135S 121205 PFBS Concentration (n g /m L ) 67.4 69.9 6 8 .7 ng/m L 3.6% PFHS Concentration (n g /m L ) 711 728 720 ng/m L 2.4% PFOS Concentration (n g /m L ) 1100 1100 1100 ng/m L 0.0% 3M LIMS ID Description GLP10-01 -02-38-029 DAL-GW-135S-0-121205 GLP10-01 -02-38-030 DAL-GW-135S-DB-121205 Average Concentration (ng/mL) %RPD Samples diluted 1:10. Table 24. DAL GW 135L 121205 PFBS Concentration (n g /m L ) 1.08 1.02 1.05 ng/m L 5.7% PFHS Concentration (n g /m L ) 10.6 10.3 10.5 ng/m L 2.9% PFOS Concentration (n g /m L ) 7.69 8.43 8.06 ng/m L 9.2% 3M LIMS ID Description GLP10-01 -02-38-031 DAL-GW-135L-0-121205 GLP10-01 -02-38-032 DAL-GW-135L-DB-121205 Average Concentration (ng/mL) %RPD Samples diluted 1:10. PFBS Concentration (n g /m L ) 7.20 6.87 7.04 ng/m L 4.7% PFHS Concentration (n g /m L ) 50.7 48.7 49.7 ng/m L 4.0% PFOS Concentration (n g /m L ) 47.2 44.8 46.0 ng/m L 5.2% Page 25 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Table 25. DAL GW 136S 121130 3M LIMS ID Description GLP10-01 -02-38-035 GLP10-01 -02-38-036 GLP10-01 -02-38-037 DAL-GW -136S-0-121130 DAL-GW -136S-DB-121130 DAL-GW -136S-FMS-121130 Average Concentration (ng/mL) %RPD PFBS PFHS PFOS Concentration (n g /m L ) % R ecovery 2.61 2.45 88.8 NA NA 86.6 2.53 ng/m L 6.3% Concentration (n g /m L ) %Recovery 19.1 18.4 114 NA NA 96.6 18.8 ng/m L 3.7% Concentration (n g /m L ) %Recovery 111 NA 118 NA 173 58.5 (1) 115 ng/m L 6 .1 % (2) NA = Not Applicable Samples diluted 1:10. (1) FMS did not meet acceptance criteria of 100 30%. (2) Method uncertainty has been expanded to 42% for PFOS. Table 26. DAL GW 136L 121130 3M LIMS ID Description GLP10-01 -02-38-038 DAL-GW-136L-0-121130 GLP10-01 -02-38-039 DAL-GW -136L-DB-121130 Average Concentration (ng/mL) %RPD Samples diluted 1:10. PFBS Concentration (n g /m L ) 2.05 2.43 2.24 ng/mL 17% PFHS Concentration (n g /m L ) 14.7 15.3 15.0 ng/m L 4.0% PFOS Concentration (n g /m L ) 89.5 94.8 92.2 ng/m L 5.8% Page 26 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Table 27. DAL GW 137S 121205 3M LIMS ID Description GLP10-01-02-38-042 GLP10-01 -02-38-043 GLP10-01-02-38-044 D A L-G W -137S -0-121205 D AL-G W -137S-D B-121205 DAL-GW-137S-F MS-121205 Average Concentration (ng/mL) %RPD PFBS PFHS PFOS Concentration (n g /m L ) % R ecovery 0.296 0.292 2.05 NA NA 87.8 0.294 ng/m L 1.4% Concentration (n g /m L ) %Recovery 0.203 0.202 2.05 NA NA 92.4 0.203 ng/mL 0.49% Concentration (n g /m L ) %Recovery 1.04 0.972 2.73 NA NA 93.0 1.01 ng/m L 6.8% NA = Not Applicable Table 28. DAL GW 137L 121205 3M LIMS ID Description GLP10-01 -02-38-045 GLP10-01-02-38-046 D AL-G W -137L-0-121205 D AL-G W -137L-D B-121205 Average Concentration (ng/mL) %RPD Table 29. DAL GW 138S 121205 PFBS Concentration (n g /m L ) 0.0794 0.0734 0.0764 ng/m L 7.9% PFHS Concentration (n g /m L ) 0.0877 0.0959 0.0918 ng/m L 8.9% PFOS Concentration (n g /m L ) 1.42 1.39 1.41 ng/m L 2.1% 3M LIMS ID Description GLP10-01 -02-38-049 DAL-GW-138S-0-121205 GLP10-01 -02-38-050 DAL-GW-138S-DB-121205 Average Concentration (ng/mL) %RPD Samples diluted 1:10. PFBS Concentration (n g /m L ) 4.87 4.75 4.81 ng/m L 2.5% PFHS Concentration (n g /m L ) 22.1 22.6 22.4 ng/m L 2.2% PFOS Concentration (n g /m L ) 233 228 231 ng/m L 2.2% Page 27 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Table 30. DAL GW 138L 121205 3M LIMS ID Description GLP10-01 -02-38-049 DAL-GW-138S-0-121205 GLP10-01 -02-38-050 DAL-GW-138S-DB-121205 Average Concentration (ng/mL) %RPD PFBS Concentration (n g /m L ) <0.0250 <0.0250 <0.0250 ng/mL PFHS PFOS Concentration (n g /m L ) Concentration (n g /m L ) 0.0551 0.0591 0.0571 ng/m L 7.0% 0.370 0.390 0.380 ng/mL 5.3% Table 31. DAL GW CW26C 121205 3M LIMS ID Description GLP10-01 -02-38-053 DAL-GW -CW 26C-0-121205 GLP10-01 -02-38-054 DAL-GW-CW26C-DB-121205 Average Concentration (ng/mL) %RPD Samples diluted 1:100. Table 32. DAL GW CW26L 121205 PFBS Concentration (n g /m L ) 4.62 4.82 4.72 ng/m L 4.2% PFHS Concentration (n g /m L ) 49.9 50.3 50.1 ng/m L 0.80% PFOS Concentration (n g /m L ) 382 385 384 ng/m L 0.78% 3M LIMS ID Description GLP10-01 -02-38-055 DAL-GW-CW26L-0-121205 GLP10-01 -02-38-056 DAL-GW-CW26L-DB-121205 Average Concentration (ng/mL) %RPD Samples diluted 1:100. PFBS Concentration (n g /m L ) 11.3 12.2 11.8 ng/m L 7.7% PFHS Concentration (n g /m L ) 95.2 95.7 95.5 ng/m L 0.52% PFOS Concentration (n g /m L ) 930 890 910 ng/m L 4.4% Page 28 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Table 33. DAL GW 140R 121130 3M LIMS ID Description GLP10-01 -02-38-057 DAL-GW -140R-0-121130 GLP10-01 -02-38-058 DAL-GW-140R-DB-121130 Average Concentration (ng/mL) %RPD Samples diluted 1:10. Table 34. DAL GW 141R 121130 PFBS Concentration (n g /m L ) 1.43 1.46 1.45 ng/m L 2.1% PFHS Concentration (n g /m L ) 15.8 15.2 15.5 ng/m L 3.9% PFOS Concentration (n g /m L ) 158 141 150 ng/mL 11% 3M LIMS ID Description GLP10-01 -02-38-059 DAL-GW -141R-0-121130 GLP10-01 -02-38-060 DAL-GW-141R-DB-121130 GLP10-01 -02-38-061 DAL-GW -141R-F MS-121130 Average Concentration (ng/mL) %RPD PFBS PFHS PFOS Concentration Concentration (n g /m L ) % R ecovery (n g /m L ) %Recovery 0.0467 NA 0.0565 NA 1.83 88.9 0.0516 ng/m L 19% 0.338 NA 0.340 NA 2.05 85.6 0.339 ng/mL 0.59% Concentration (n g /m L ) %Recovery 6.25 NA 5.91 NA 8.03 NC 6.08 ng/m L 5.6% NA = Not Applicable NC = Not Calculated; Spike level was less than 0.5x the endogenous sample concentration. Page 29 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Table 35. DAL GW 142R 121130 3M LIMS ID Description GLP10-01-02-38-062 GLP10-01 -02-38-063 DAL-GW -142R-0-121130 D A L-G W -142R -D B -121130 Average Concentration (ng/mL) %RPD Samples diluted 1:10. Table 36. DAL GW 145R 121204 PFBS Concentration (n g /m L ) 1.72 1.53 1.63 ng/m L 12% PFHS Concentration (n g /m L ) 9.45 9.15 9.30 ng/m L 3.2% PFOS Concentration (n g /m L ) 110 99.8 1 0 5 ng/m L 9.7% 3M LIMS ID Description GLP10-01 -02-38-067 DAL-GW-145R-0-121204 GLP10-01 -02-38-068 DAL-GW-145R-DB-121204 GLP10-01 -02-38-069 DAL-GW-145R-F MS-121204 Average Concentration (ng/mL) %RPD PFBS PFHS PFOS Concentration Concentration (n g /m L ) % R ecovery (n g /m L ) %Recovery 1.62 NA 1.55 NA 79.8 78.5 1.59 ng/m L 4.4% 7.51 NA 7.34 NA 95.5 89.3 7.43 ng/m L 2.3% Concentration (n g /m L ) %Recovery 116 NA 120 NA 205 87.0 118 ng/m L 3.4% NA = Not Applicable Samples diluted 1:10. Page 30 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Table 37. Trip Blank 3M LIMS ID GLP10-01 -02-38-072 GLP10-01 -02-38-073 GLP10-01 -02-38-074 GLP10-01 -02-38-075 Description D A L-G W -TR IP 01-0DAL-GW-TRIP01 -LSDAL-GW-TRIP01 -MSDAL-GW-TRIP01 -HS- NA = Not Applicable Sample GLP10-01-02-38-075 diluted 1:10. Table 38. Rinseate Blanks PFBS PFHS PFOS Concentration (n g /m L ) <0.0250 1.79 16.2 89.2 %Recovery NA 89.5 81.3 89.6 Concentration (n g /m L ) <0.0250 1.87 17.5 84.8 %Recovery NA 93.5 88.7 8 6 .0 Concentration (n g /m L ) <0.0278 1.79 19.9 82.8 %Recovery NA 96.5 99.5 82.8 3M LIMS ID GLP10-01 -02-38-070 GLP10-01 -02-38-071 Description D AL-G W -131S-R B-121204 D A L-G W -145R -R B -121204 PFBS Concentration (n g /m L ) <0.0250 <0.0250 PFHS Concentration (n g /m L ) <0.0250 <0.0250 PFOS Concentration (n g /m L ) <0.0278 <0.0278 Page 31 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 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. 12 Attachments Attachment A: Protocol Amendment 38 (General Project Outline) Attachment B: Representative Chromatograms and Calibration Curves Attachment C: Analytical Method - ETS-8-044.1 Attachment D: Method Deviation Page 32 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS In Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 13 Signatures Cleston Lange, Ph.D., 3M Principal Analytical Investigator '* 4/ William K. Reagen, Ph.D., 3M Environmental Laboratory Department Manager Date Date Page 33 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Attachment A: Protocol A mendm ent Page 34 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Analytical Protocol: GLP10-01-02 Amendment 38 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. 38 Amendment Date: November 27, 2012 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 6 Page 35 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Analytical Protocol: GLP10-01-02 Amendment 38 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 w ording o f the protocol are required. Amend to read: No changes to the w ording o f the protocol are required This am e nd m ent only ad dre sses and docum ents the addition of the G eneral P roject O utline (G P O ) fo r the collection and analysis o f g ro un dw a te r sam ples from D ecatur, AL, and conducted as part o f the 3M D ecatur Phase 3 P rogram for PFO S, PFHS and PFBS (G LP 10-01-02). T he an ticip ated sam ple collection will o ccu r around the tim e fra m e d u rin g the w e e k o f D ecem ber 3, 2012. T he g ro un dw a te r sam ples for this sam pling e ve n t w ill be e n tered into the 3M E nvironm ental Laboratory LIM S as project G LP10-01 -02-38 and reported as interim report G LP 10-01-02 38, (reflecting study G LP 10-01-02 and am endm ent -38). Reason. T h e re a s o n fo r th is a m e n d m e n t is to d o c u m e n t th e G e n e ra l P ro je c t O u tlin e (G P O ) w h ic h d e s c rib e s the anticipate groundw ater sam ple collection event to be conducted for the 3M D ecatur, A L facility T h e G P O is th re e p a g e s in length a n d in clud ed as attach ed to this a m e n d m e n t form . Page 2 of 6 Page 36 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Analytical Protocol: GLP10-01-02 Amendment 38 Amendment Approval W illiam Reagen, EHS O pns Environm ental Lab M anagem ent 7n .A Jaisim ha Kesri P.E , DEE, Study Director C D a te D a te Page 3 of 6 Page 37 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Analytical Protocol: GLP10-01-02 Amendment 38 3 R A Environmental Health & Safety Operations, Environmental Laboratory General Project Outline To: G a ry H o he nstein , 3M E H S & O p n s From: S usa n W o lf, 3M E H S & O p ns; E nviron m ental Lab cc: W illiam Reagen, 3M EHS&O pns; Environm ental Lab Cieston Lange, 3M EHS&O pns; Environm ental Lab Jai Kesari, W eston Solutions Date: N o v e m b e r 27, 2012 Subject: A nalysis of P erflucrooctane S ulfonate (PFO S), P erfluorohexane S ulfonate (P F H S ) and P e rflu o ro b u ta n e su lfo n a te (P F B S ) in G ro u n d w a te r, S oil and S e d im e n t fo r the 3M D e ca tu r P hase 3 S ite-R elated M onitoring P rogram ; G LP Interim R eport 38 - F ortne r S ludge Incorporation A rea (FSIA) G roun dw ater - D ecem ber 2012 S am pling 1 General Project Information C o n ta c ts Lab Request Number Six Digit Department Number Project Schedule/Test Dates 3M Sponsor Representative Gary Hohenstein 3M EHS Operations 3M Building 224-5W-03 Saint Paul, MN 55144-1000 Phone: (651) 737-3570 aahohenstein@mmm.coinn 3M Environm ental Laboratory Management William K. Reagen 3M EHS Opns, Environmental Laboratory 250-5N-17 651 733-9739 wkreaaen mmm.com Principal Analytical Investigator Cieston Lange 3M EHS Opns, Environmental Laboratory 260-5N-17 651 733-9860 cdanae@mmm.com Sam pling C oordinator Timothy Frlnak Weston Solutions Tim othv.fT !nak@ w estonso!utions,com Phone: (334)-332-912~3 G L P 1 0-01-02-38 Dept #530711, Project #0022674449 Sampling scheduled fo r the week of December 3, 2012 All verbal and written correspondence will be directed to Gary Hohenstein. Page 4 of 6 Page 38 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Analytical Protocol: GLP10-01-02 Amendment 38 2 Background Information and Project Objective(s) The 3M EH S O pe ra tion s Laboratory (3M E nvironm ental Lab) will receive and an alyze g ro un dw a te r sam ples collected from an expected thirty-one g ro un dw a te r w ells fo r P erflu oro buta nesu lfon ate (P FB S ), P erflu oro hexa ne sulfo nate (P FH S ), and P erflu oro octan esulfo na te (P F O S ) from the F o rm e r Sludge Incorporation A rea (FS IA ). A nalyses w ill be conducted under the G LP req uire m en ts of E P A T S C A G ood Laboratory P ractice S tandards 40 C FR 792, G roun dw ater sam ples from D ecatur, A L w ill be collected by W eston S olutions personnel th e w eek of D ecem ber 3, 2 0 1 2 T he final report w ill be subm itted to G ary H ohenstein and Jai K esari upon com pletion under interim report G L P 1 0-01-02-38. 3 Project Schedule _________ ______________ __ S am ple collection bottles will be prepared by 3M Environm ental Laboratory fo r sam pling the w e ek o f D ecem ber 3 ,2 0 1 2 . S am ple bottles will be shipped in coolers to 3M D ecatur fo r arrival by Friday, N o vem b er 30, 2012. Sam ple bottles should be stored refrigerated on-site until sam ple collection. M artin S m ith \ W eston T railer 3M D ecatur P lant 1400 S tate D ocks Road D ecatur, A labam a 35601 4 Test Parameters The targeted lim it o f quantitation will be 0.025 ng/mL (ppb) fo r PFBS, PFHS, and PFOS. A total of thirty-two sam pling locations have been specified. Based on previous sam pling o f the FSIA (G LP1001-02-07, GLP1Q-01-02-011, G LP 10-01-02-18, GLP 10-01-02-24, and G LP 10-01-02-28), m ore than half o f the sam pling locations are expected to have PFBS, PFHS, or PFOS levels greater than 100 n g /m L T he 3M Environm ental Laboratory has analyzed these locations m ultiple tim es and has dem onstrated that the m ethod used fo r th e a n a lysis o f th e sa m p le s Is ap pro pria te fo r th e sa m p le m a trix (field m a trix sp ike reco verie s ha ve dem onstrated th a t the m ethod is accurate fo r PFBS, PFHS, and P F O S in the given sam p le m atrix). Therefore, for this sam pling event, field m atrix spikes will only be prepared on select sam pling locations. If need be, laboratory m atrix spikes m ay be prepared. For each sam pling location, a sam ple and sam ple duplicate will be collected. Table 1 indicates the sam pling locations in clud ed in th is sa m p lin g event. T a b le 2 in dica tes the sam p lin g lo ca tio n s fo r w h ich a field m atrix spike will be included. T he "fill to here" line on each 250 m L Nalgene bottle will be 200 m L O ne sets o f trip blanks consisting o f reagent-grade water, a low-level trip blank spike, m id-level field spike, and a high-level trip blank spike will be prepared at the 3M E nvironm ental Laboratory and sent to the sam pling location with the other bottles Tw o additional bottles will be included to be used fo r the preparation o f the eq uipm en t rinseate blanks. A 1-L bottle of laboratory reagent w ater w ill be provided to be used to collect the rinseate blanks. Page 5 of 6 Page 39 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Analytical Protocol: GLP10-01-02 Amendment 38 Table 1. Sampling Locations fo r this Sam pling Event. 130R 130S 130L 131R 131S 131L 133R 1333 133L 134R 134S 134L 135R 135S 135L 136R 136S 136L FSIA 137R 137S 137L 138R 138S 13SL CW 26C CW 26L 140R 141 R 142R 143R 145R Table 2. Sampling Locations with Field Matrix Spike Sample Well No. Spike Cone. (ng/mL) 137S and 141R 131S and 143R 2.0 20 130S, 136S, and 145R T rip Blank 100 2.0 20 100 5 Test Methods Sam ples m ay be analyzed by ETS-8-044 1 "M ethod o f Analysis for the D eterm ination o f Periluorinated C o m p o u n d s In W a te r by LC /M S /M S ; D ire ct Injection A na lysis". T h is m e th o d is a d ire c t in je ctio n m e th o d w h e re sam ples w ould be analyzed as neat aqueous sam ple o r as diluted aqueous sam ples. Laboratory control sam ples prepared with the sam ples m ust have an average recovery w ithin 100+20% and a R S D <20% . T h e data quality o b je ctive fo r th is stu d y is q u a n tita tive resu lts fo r th e ta rg e t a n a lyte s w ith an analytical accuracy o f 10030% . Field m atrix spikes not yielding recoveries w ithin 10030% will be addressed in th e rep ort a n d th e fin a l a ccu ra cy s ta te m e n t m a y be ad ju ste d acco rdingly. Internal standards and/or surrogates m ay be added to the sam ples after sam ple collection and prior to sam ple analysis, to aid in the data quality objectives. 6 Reporting Requirements ______ _ _ ______ For each sam pling location, the report will contain the results for the sam ple, sam ple duplicate, and field m atrix spike. Trip blank and trip blank spikes will be reported for the sam pling event as will a n y equipm ent/rinseate blan ks p re pa red in th e field. La b o ra to ry con tro l spikes o f rea ge nt w a te r p re pa red a t th e tim e o f sa m p le preparation will also be reported and used to evaluate the overall m ethod accuracy and precision M ethod blanks of reagent w ater prepared at the tim e o f sam ple preparation will be used to determ ine the m ethod detection limit. Page 6 of 6 Page 40 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Attachment B: Representative Sa m ple Chrom atograms and Calibration Curve(s) Page 41 of 109 Workstation: W0119058 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Batch Name: b121226a.dab *Data printed by KJU Printing Time: 3:09:12 PM P r i n t i n g Date: Friday, D e c e m b e r 28, 2012 Page 42 of 109 Workstation: W0119058 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Batch Name: b121226a.dab b121226a.rdb (PFBS): "Quadratic" Regression ("1 / x" weighting): y = -1.77e+003 xA2 + 3.37e+005 x + 3.44e+003 (r = 0.9998) ______________________________________________________________________________ Concentration, ng/mL_________________________ *Data printed by KJU Page 1 of 1 Printing Time: 3:09:30 PM P r i n t i n g Date: Friday, D e c e m b e r 28, 2012 Page 43 of 109 *** Buster J2930203 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Batch Name: b121226a.dab Data printed by STW Printing Time: 10:15:00 AM P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 44 of 109 *** Buster J2930203 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Batch Name: b121226a.dab Data printed by STW Printing Time: 10:15:00 AM P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 2 of 20 Page 45 of 109 *** Buster J2930203 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Batch Name: b121226a.dab Data printed by STW Printing Time: 10:15:01 AM P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 3 of 20 Page 46 of 109 *** Buster J2930203 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Batch Name: b121226a.dab Data printed by STW Printing Time: 10:15:01 AM P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 4 of 20 Page 47 of 109 *** Buster J2930203 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Batch Name: b121226a.dab Data printed by STW Printing Time: 10:15:02 AM P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 5 of 20 Page 48 of 109 *** Buster J2930203 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Batch Name: b121226a.dab Data printed by STW Printing Time: 10:15:03 AM P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 6 of 20 Page 49 of 109 *** Buster J2930203 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Batch Name: b121226a.dab Data printed by STW Printing Time: 10:15:03 AM P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 7 of 20 Page 50 of 109 *** Buster J2930203 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Batch Name: b121226a.dab Data printed by STW Printing Time: 10:15:03 AM P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 8 of 20 Page 51 of 109 *** Buster J2930203 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Batch Name: b121226a.dab Data printed by STW Printing Time: 10:15:04 AM P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 9 of 20 Page 52 of 109 *** Buster J2930203 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Batch Name: b121226a.dab Data printed by STW Printing Time: 10:15:04 AM P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 53 of 109 *** Buster J2930203 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Batch Name: b121226a.dab Data printed by STW Printing Time: 10:15:04 AM P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 11 of 20 Page 54 of 109 *** Buster J2930203 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Batch Name: b121226a.dab Data printed by STW Printing Time: 10:15:05 AM P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 12 of 20 Page 55 of 109 *** Buster J2930203 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Batch Name: b121226a.dab Data printed by STW Printing Time: 10:15:05 AM P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 56 of 109 *** Buster J2930203 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Batch Name: b121226a.dab Data printed by STW Printing Time: 10:15:05 AM P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 57 of 109 *** Buster J2930203 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Batch Name: b121226a.dab Data printed by STW Printing Time: 10:15:06 AM P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 15 of 20 Page 58 of 109 Buster J2930203 ISample Name: "b121226a083" Sample ID: "GLP10-01-02-38-027" File: "b121226a.wiff" Peak Name: "PFHS" Mass(es): "399.000/99.000 Da,399.000/80.000 Da" Comment: "DAL-GW-135R-0-" Annotation: "" ample Index: 83 ample Type: Unknown :alculat ng/mL oc. Algorithm: Specify dth dth Typ : 13.9 2312168 6. 45e+005 Sample Name: "b121226a087" Sample ID: "GLP10-01-02-38-031" File: Peak Name: "PFHS" Mass(es): "399.000/99.000 Da,399.000/80.000 Da" Comment: "DAL-GW-135L-0-" Annotation: "" ample Index: 87 ample Type: Unknown :alculat 50. ' ng/m 12/27/2012 3:14:26 PM Algorithm: Specify I [eight: lidth: idth: 0.00 0.00 3 30.0 s. Typ i Time: 13.9 1674073 coi 4.36e+0 05 13.94 lime, min GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Batch Name: b121226a.dab I Sample Name: "b121226a085" Sample ID: "GLP10-01-02-38-029" File: 'b121226a.wiff" I Peak Name: "PFHS" Mass(es): "399.000/99.000 Da.399.000/80.000 Da" J Comment: "DAL-GW-135S-0-" Annotation: "" ample Index: 85 ample Type: Unknown alculated Conc: 10.6 ng/mL cq. Date: 12/27/2012 cq. Time: 2:34:17 PM Iodified: 1.00e5 9.50e4 9.00e4 8.50e4 >othing Width: 8.00e4 7.50e4 7.00e4 : 13.9 361379 coi 1.05e+005 6.50e4 6.00e4 5.50e4 5.00e4 4.50e4 4.00e4 3.50e4 3.00e4 2.50e4 2.00e4 1.50e4 1.00e4 5000.00 0.00 Sample Name: "b121226a092" Sample ID: "GLP10-01-02-38-035" File: Peak Name: "PFHS" Mass(es): "399.000/99.000 Da,399.000/80.000 Da' Comment: "DAL-GW-136S-0-" Annotation: "" :ample Index: 92 ample Type: Unknown :alculated Conc: 19.1 ng/mL icq. Date: 12/27/2012 icq. Time: 4:54:57 PM 17e5' oc. Algorithm: Specify 3.94 6 8 10 12 14 16 18 Time, min Data printed by STW Printing Time: 10:15:06 AM Time, min P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 16 of 20 6 8 10 12 14 16 18 Time, min Page 59 of 109 *** Buster J2930203 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Batch Name: b121226a.dab Data printed by STW Printing Time: 10:15:06 AM P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 17 of 20 Page 60 of 109 *** Buster J2930203 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Batch Name: b121226a.dab Data printed by STW Printing Time: 10:15:07 AM P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 18 of 20 Page 61 of 109 *** Buster J2930203 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Batch Name: b121226a.dab Data printed by STW Printing Time: 10:15:07 AM P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 19 of 20 Page 62 of 109 *** Buster J2930203 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Batch Name: b121226a.dab Data printed by STW Printing Time: 10:15:07 AM P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 20 of 20 Page 63 of 109 Workstation: W0119058 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Results Name: k130103a.rdb Data printed by KJU Printing Time: 10:32:18 AM P r i n t i n g Date: M onday, J a n u a r y 07, 2013 Page 64 of 109 *ETS-Kirk GLP10-01-02; Interim Report 38 Analysis o f PFBS, PFHS, and PFOS in G roundwater Sam ples Former Sludge Incorporation Area (FSIA); Decatur, AL Results Name: k130103a.rdb *Data printed by STW P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 65 of 109 *ETS-Kirk GLP10-01-02; Interim Report 38 Analysis o f PFBS, PFHS, and PFOS in G roundwater Sam ples Former Sludge Incorporation Area (FSIA); Decatur, AL Results Name: k130103a.rdb *Data printed by STW P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 2 of 20 Page 66 of 109 *ETS-Kirk GLP10-01-02; Interim Report 38 Analysis o f PFBS, PFHS, and PFOS in G roundwater Sam ples Former Sludge Incorporation Area (FSIA); Decatur, AL Results Name: k130103a.rdb *Data printed by STW P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 67 of 109 *ETS-Kirk GLP10-01-02; Interim Report 38 Analysis o f PFBS, PFHS, and PFOS in G roundwater Sam ples Former Sludge Incorporation Area (FSIA); Decatur, AL Results Name: k130103a.rdb *Data printed by STW P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 4 of 20 Page 68 of 109 *ETS-Kirk GLP10-01-02; Interim Report 38 Analysis o f PFBS, PFHS, and PFOS in G roundwater Sam ples Former Sludge Incorporation Area (FSIA); Decatur, AL Results Name: k130103a.rdb *Data printed by STW P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 69 of 109 *ETS-Kirk GLP10-01-02; Interim Report 38 Analysis o f PFBS, PFHS, and PFOS in G roundwater Sam ples Former Sludge Incorporation Area (FSIA); Decatur, AL Results Name: k130103a.rdb *Data printed by STW P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 6 of 20 Page 70 of 109 *ETS-Kirk GLP10-01-02; Interim Report 38 Analysis o f PFBS, PFHS, and PFOS in G roundwater Sam ples Former Sludge Incorporation Area (FSIA); Decatur, AL Results Name: k130103a.rdb P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 71 of 109 *ETS-Kirk GLP10-01-02; Interim Report 38 Analysis o f PFBS, PFHS, and PFOS in G roundwater Sam ples Former Sludge Incorporation Area (FSIA); Decatur, AL Results Name: k130103a.rdb *Data printed by STW P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 8 of 20 Page 72 of 109 *ETS-Kirk GLP10-01-02; Interim Report 38 Analysis o f PFBS, PFHS, and PFOS in G roundwater Sam ples Former Sludge Incorporation Area (FSIA); Decatur, AL Results Name: k130103a.rdb *Data printed by STW P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 9 of 20 Page 73 of 109 *ETS-Kirk GLP10-01-02; Interim Report 38 Analysis o f PFBS, PFHS, and PFOS in G roundwater Sam ples Former Sludge Incorporation Area (FSIA); Decatur, AL Results Name: k130103a.rdb *Data printed by STW P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 74 of 109 *ETS-Kirk GLP10-01-02; Interim Report 38 Analysis o f PFBS, PFHS, and PFOS in G roundwater Sam ples Former Sludge Incorporation Area (FSIA); Decatur, AL Results Name: k130103a.rdb *Data printed by STW P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 11 of 20 Page 75 of 109 *ETS-Kirk GLP10-01-02; Interim Report 38 Analysis o f PFBS, PFHS, and PFOS in G roundwater Sam ples Former Sludge Incorporation Area (FSIA); Decatur, AL Results Name: k130103a.rdb *Data printed by STW P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 12 of 20 Page 76 of 109 *ETS-Kirk GLP10-01-02; Interim Report 38 Analysis o f PFBS, PFHS, and PFOS in G roundwater Sam ples Former Sludge Incorporation Area (FSIA); Decatur, AL Results Name: k130103a.rdb *Data printed by STW P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 13 of 20 Page 77 of 109 *ETS-Kirk GLP10-01-02; Interim Report 38 Analysis o f PFBS, PFHS, and PFOS in G roundwater Sam ples Former Sludge Incorporation Area (FSIA); Decatur, AL Results Name: k130103a.rdb *Data printed by STW P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 78 of 109 *ETS-Kirk ISample Name: "k130103a065" Sample ID: "GLP10-01-02-38-013" File: "k130103a.wiff" Peak Name: "PFOS" Mass(es): "499.000/80.000 Da,499.000/99.000 Da,499.000/130.000 Da" Comment: "DAL-GW-131L-0-" Annotation: "" ample Index: 65 ample Type: Unknown :alculat 39. i 4/2 013 25:27 ng/mL 2.0e5 Algorithm: Specify I dth dth Typ 1378029 c 2.02e+ 005 9.0e4 8.0e4 7.0e4 6.0e4 5.0e4 4.0e4 3.0e4 2.0e4 12.0 12.5 Sample Name: "k130103a069" Sample ID: "GLP10-01-02-38-017" File: "k130103a.wiff" Peak Name: "PFOS" Mass(es): "499.000/80.000 Da,499.000/99.000 Da,499.000/130.000 Da" Comment: "DAL-GW-133S-0-" Annotation: "" ample Index: 69 ample Type: Unknown alculat 193. ng/m 1.05e6 Algorithm: Specify P 1.00e6 9.50e5 9.00e5 8.50e5 dth dth 8.00e5 7.50e5 Typ i Time: 14.3 7850747 c 1. 09e+006 7.00e5 6.50e5 6.00e5 5.50e5 5.00e5 4.50e5 4.00e5 3.50e5 3.00e5 2.50e5 2.00e5 1.50e5 1.00e5 5.00e4 0.00 *Data printed by STW P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 14.27 14.40 6^ 14.27 GLP10-01-02; Interim Report 38 Analysis o f PFBS, PFHS, and PFOS in G roundwater Sam ples Former Sludge Incorporation Area (FSIA); Decatur, AL Results Name: k130103a.rdb Sample Name: "k130103a067" Sample ID: "GLP10-01-02-38-015" File: "k130103a.wiff" Peak Name: "PFOS" Mass(es): "499.000/80.000 Da,499.000/99.000 Da,499.000/130.000 J Comment: "DAL-GW-133R-0-" Annotation: "" ample Index: 67 ample Type: Unknown oncentration: alculated Conc: N/A 44.1 ng/mL 2 7e5 27e5 cq. Date: 1/4/2013 2 6e5 cq. Time: 9:05:06 AM odified: Yes 2.5e5 roc. Algorithm: SpecifyParameters - MQ III 2.4e5 oise Iercentage: 90 ase. Sub. Window: 1. 00 min 23e5 "Itt iN N et 6Yes 22e5 in. Peak Height: 0.00 cps 2 1e5 -- n .00 sec >othing Width: points 2.0e5 12.0 Sample Name: "k130103a071" Sample ID: "GLP10-01-02-38-019" File: "k130103a.wiff" I Peak Name: "PFOS" Mass(es): "499.000/80.000 Da,499.000/99.000 Da,499.000/130.000 Da" Comment: "DAL-GW-133L-0-" Annotation: "" ample Type: :alculated C N/A 455. '4/2013 g/ oc. Algorithm: Specify 13.5 14.0 14.5 15.0 15.5 16.0 16.5 Time, min 14.26 14.39 14 14.5 15.0 15.5 16.0 16.5 Time, min 12.0 12.5 Page 15 of 20 1 Page 79 of 109 *ETS-Kirk ISample Name: "k130103a073" Sample ID: "GLP10-01-02-38-021" File: "k130103a.wiff" Peak Name: "PFOS" Mass(es): "499.000/80.000 Da,499.000/99.000 Da,499.000/130.000 Da" Comment: "DAL-GW-134R-0-" Annotation: "" ample Index: 73 ample Type: Unknown :alculat 904 /2013 03:57 ng/mL 5.2e5 5.0e5 Algorithm: Specify I ath dth Typ 036235 c 5.39e+005 3.8e5 3.6e5 3.4e5 3.2e5 3.0e5 2.8e5 2.6e5 2.4e5 2.2e5 2.0e5 14.40 15v. 12.0 12.5 13.0 ISample Name: "k130103a080" Sample ID: "GLP10-01-02-38-025" File: "k130103a.wiff" Peak Name: "PFOS" Mass(es): "499.000/80.000 Da,499.000/99.000 Da,499.000/130.000 Da" Comment: "DAL-GW-134L-0-" Annotation: "" ample Type: Unknown lalculat ng/m 22:43 PM 3.2e5 oc. Algorithm: Specify 13.5 ; Height: ; Width: j Width: Typ i Time: 14 .1 2406763 coi 3.31e+005 lime, min 15.5 16.0 GLP10-01-02; Interim Report 38 Analysis o f PFBS, PFHS, and PFOS in G roundwater Sam ples Former Sludge Incorporation Area (FSIA); Decatur, AL Results Name: k130103a.rdb I Sample Name: "k130103a078" Sample ID: "GLP10-01-02-38-023" File: "k130103a.wiff" I Peak Name: "PFOS" Mass(es): "499.000/80.000 Da,499.000/99.000 Da,499.000/130.000 Da" J Comment: "DAL-GW-134S-0-" Annotation: "" ple Typ 1/4/2013 12:43:02 ng/ 13.91 Smoothing Width: RT Window: 30.0 ie: 13.9 5113164 coi 7.80e+005 1402 1I.8K 12.0 12.5 Sample Name: "k130103a082" Sample ID: "GLP10-01-02-38-027" File: "k130103a.wiff" Peak Name: "PFOS" Mass(es): "499.000/80.000 Da,499.000/99.000 Da,499.000/130.00( Comment: "DAL-GW-135R-0-" Annotation: "" lample Index: 82 ample Type: Unknown :alculated Conc: 1100. ng/mL icq. Date: 1/4/2013 icq. Time: 2:02:19 PM 6.0e5 Iodified: Yes Specify 13.0 13.5 15.0 15.5 16.0 16.5 lime, mir 6.34e+005 13.84 12.0 12.5 13.0 13.5 *Data printed by STW P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 lime, min 15.5 16.0 12.0 12.5 13.0 Page 16 of 20 13.5 15.0 15.5 16.0 16.5 lime, min Page 80 of 109 *ETS-Kirk GLP10-01-02; Interim Report 38 Analysis o f PFBS, PFHS, and PFOS in G roundwater Sam ples Former Sludge Incorporation Area (FSIA); Decatur, AL Results Name: k130103a.rdb *Data printed by STW P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 17 of 20 Page 81 of 109 *ETS-Kirk GLP10-01-02; Interim Report 38 Analysis o f PFBS, PFHS, and PFOS in G roundwater Sam ples Former Sludge Incorporation Area (FSIA); Decatur, AL Results Name: k130103a.rdb *Data printed by STW P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 18 of 20 Page 82 of 109 *ETS-Kirk GLP10-01-02; Interim Report 38 Analysis o f PFBS, PFHS, and PFOS in G roundwater Sam ples Former Sludge Incorporation Area (FSIA); Decatur, AL Results Name: k130103a.rdb *Data printed by STW P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 19 of 20 Page 83 of 109 *ETS-Kirk GLP10-01-02; Interim Report 38 Analysis o f PFBS, PFHS, and PFOS in G roundwater Sam ples Former Sludge Incorporation Area (FSIA); Decatur, AL Results Name: k130103a.rdb *Data printed by STW P r i n t i n g Date: Thursday, J a n u a r y 17, 2013 Page 20 of 20 Page 84 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Attachment C: A nalytical Meth o d (s) Page 85 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 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 j 7/ K Approved By: William K. Reagen, Technical Director, Environmental Laboratory J / J Q i/ // 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 86 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 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). Water 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 Water 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 E 11-0667. 2 Guidance for establishing method QC Criteria based on a.) FDA May 2 0 0 1 , "Guidance for Industry, Bioanalytical Method Validation", b.) EPA Method 5 37, and c.) European Commission: Guidance for Generating and Reporting Methods of 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 87 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 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 Water 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 groundwater:MeOH. 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 88 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 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 89 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 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/Procedural 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 90 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 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 0.0001g 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 Water - 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 91 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 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 (C10 Perfluorinated Acid) PFUnA, Perfluoroundecanoic Acid, (C^ Perfluorinated Acid) PFDoA, Perfluorododecanoic Acid, (C12 Perfluorinated Acid) PFTrDA, Perfluorotridecanoic Acid, (C13 Perfluorinated 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-isotopically 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- C4]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 -13C2 ]PFDA, [1,2,3,4,5,6,7 -13C7]PFUnA, [1,2 -13C2]PFDoA, [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]PFOSA (Wellington Laboratories, Guelph, ON) in combination with added ([1,2,3,4,5-13C5]PFPeA, ([1,2,3,4-C4]PFHpA, and [18O2]PFBS 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 92 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 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 for percent salt, acid [ETS-4-031] a n d purity) and dilute to 100 mL with methanol or other suitable solvent, in a 100 mL volumetric flask. T ransfer 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. lx x. , x molecular weight of anion salt correction factor = ---------------------- ---------------- moclecular 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 3 A 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 E11-0560. 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 93 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 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 -13C2]PFHxA, [1,2,3,4,5,6,7,8-13C8]PFOA, [1,2,3,4,5,6,7,8,9- C q]PFNA, [1,2,3,4,5,6 -13C6]PFDA, [1,2,3,4,5,6,7 -13C7]PFUnA, [1,2 13C2 ]PFDoA, [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 [18O2]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 94 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Table 1. Stable Isotope PFCAs and PFSAs used for ISs and SRSs Coim pound Name Synonym or Acronym 13C4 -Perfluorobutanoic acid [1,2,3,4-13C4]PFBA 13C4-Perfluoropentanoic acid [1,2,3,4,5-13C5]PFPeA 13C2-Perfluorohexanoic acid [1,2 -13C2]PFHxA 13C4-Perfluoroheptanoic acid [1,2,3,4-13C4]PFHpA 13C8-Perfluorooctanoic acid [1,2,3,4,5,6,7,8-13C8]PFOA 13C9-Perfluorononanoic acid [1,2,3,4,5,6,7,8,9-13C9]PFNA 13C6-Perfluorodecanoic acid [1,2,3,4,5,6 -13C6]PFDA 13C7-Perfluoroundecanoic acid [1,2,3,4,5,6,7 -13C7]PFUnA 13C2 -Perfluorododecanoic acid [1,2 -13C2]PFDoA 18O2-Ammonium Perfluorobutane sulfonate [18O2]PFBS 13C3 -Ammonium Perfluorohexane sulfonate [1,2,3-13C3]PFHS 13C8-Sodium Perfluorooctane sulfonate [1,2,3,4,5,6,7,8-13C8]PFOS 13C8-Perfluorooctanesulfonamide 13C4-Perfluorooctanoic acid [1,2,3,4,5,6,7,8-13C8]FOSA [1,2,3,4-13C4]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 13C4], 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 13C8-Perfluorooctane sulfonate [1,2 -13C2]PFUnA [1,2,3,4-13C4]PFOS SRS for all PFCAs C9-C13 Wellington 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 95 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 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 of 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 of 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 I Water, or other suitable solvent(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:MeOH. 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 96 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 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 97 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 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 98 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 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 99 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 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 100 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 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, 5p.m or 2.1 mm x 50 mm, 5p.m) 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, 5p.m (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. Liquid Chromatography Gradient 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 (pL/min) 750 750 750 750 750 750 Percent A (2 m M 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. 2.1 mm 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 101 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Table 4 Suggested MRM Transitions for Target Analytes, Surrogates, and Internal Standards Analyte PFB A (C4 Acid) PFPeA (C5 Acid) PFH xA (C6 Acid) PFH pA (C7 Acid) PFO A (C8 Acid) PFN A (C9 Acid) PFD A (C10 Acid) PFU nA (C11 Acid) PFD oA (C 12 Acid) PFTA (C13 Acid) FBSA (C4 Sulfonamide) FO SA (C8 Sulfonam ide) PFBS (C 4 Sulfonate) PFH S (C6 Sulfonate) PFO S (C8 Sulfonate) [1 ,2 ,3 ,4 -13C 4]P F B A [1,2,3,4,5 - 13C 5 ]P F P e A H ,2 - 13C 2 lP F H x A [1 ,2,3,4- 13C 4lP F H p A [1 ,2 ,3 ,4 ,5 ,6 ,7 ,8 -13C 8lP F O A H ,2 ,3 ,4 ,5 ,6 ,7 ,8 ,9 -13C 9lP F N A r 1,2,3,4,5,6 - 13C 6lP F D A r 1 ,2 ,3,4,5,6,7 - 13C 7 lP F U n A [1,2 - 13C 2 lP F D o A [18O 2lP F B S [1 ,2 ,3 -13C 3lP F H S [1 ,2,3,4- 13C 4lP F O S [ 1 ,2 ,3 ,4 ,5 ,6 ,7 ,8 -13C 8lF O S A [1 ,2 ,3 ,4 -13C 4lP F O A [1,2,3,4- ^ l P F O S [1,2 - 13C 2 lP F U n A Analyte Description T arg et T arg et T arg et T arg et T arg et T arg et T arg et T arg et T arg et T arg et T arg et T arg et T arg et T arg et T arg et IS for PFBA IS for PFPeA IS for PFHxA IS for PFH pA IS for PFO A IS for PFN A IS for PFD A IS for PFU nA IS for PFD oA and PFTA IS for PFBS IS for PFHS IS for PFOS IS for FO SA Surrogate (C4-C8 Acids) Surrogate(Sulfonates, FOSA) Surrogate (C9-C13 Acids) M ass Transition Q1 (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 (am u) 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 102 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 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 as part 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 - 100pL) 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 103 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 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 (-5^-) LCS% recovery = ----------------------------- mL * 100% ng Spike Concentration (--^-) mL Calculate the percent recovery of the LMS using the following equation: LMS % recovery ng ng LMS C oncentration (-- --) - C oncentration of Sam ple (---- ) mL mL ng Spike C oncentration (---- ) 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) ^ 1 0 0 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 Calibration 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 Performance: 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 104 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 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. Limits of Quantitation (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. Demonstration of Specificity: 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 Method/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 Control 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 Duplicates: 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 Matrix 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 105 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 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 2 002. 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 106 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL 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 107 of 109 Attachment D: Deviation(s) GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL Page 108 of 109 GLP10-01-02; Interim Report 38 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Former Sludge Incorporation Area (FSIA); Decatur, AL R ecord of D eviation/N onconform ance I. Identification Study / Project No. GLP10-01-02-38 Date(s) of Occurrence: 1/3/13 Document Number: ETS-8-044.1 Deviation type (Check one) SOP Protocol Equipment Procedure 0 Method GPO Other: II. Description (attach extra pages as needed) Method Requirements: 1. FMS recovery within 30%. 2. RPD values <20%. 3. LCS recovery within 20%._____________________________________________________________ Actual procedure/process: 1. The sampling location DAL G W 136S had a PFOS FMS recovery of 58.5%. 2. The sampling location DAL G W 130L had a sample/sample duplicate RPD of 22% for PFOS.. 3. The low level LCSs (0.2ppb) had an average recovery of 127% for PFOS.__________________ III. Actions Taken ________________________ (such a s a m e n d m e n t issued, S O P revision, e tc.)_______________________ 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 sampling location DAL G W 136S will be expanded to 42% for PFOS. The average FMS recovery for all FMS samples was calculated to determine whether the method uncertainty for all samples needed to be further expanded due to the limited number of FMS samples. Since the overall average FMS recovery met the acceptance criteria of 100 30% at 84.9% , the method uncertainty will only be expanded for DAL G W 136S. 2. The non-compliant RPDs will be flagged in the final report. 3. All LCSs were used to determine the overall analytical method uncertainty. Since the % bias of the low level LCSs (27% ) was less than the calculated analytical method uncertainty (36% ) for PFOS, the ________ method uncertainty does not need to be further expanded.___________________________________________ Actions: Halting of Work Client Notification Work Recall Withholding of Report 0 Other: Deviations will be noted in final report. Project Lead/PAI Approval: Q /v Date:, , Study Director (if GLP): Sponsor Approval (for GLP protocol deviations): NA Technical Reviewer (optional): NA Laboratory Depajjnnerii-Manager Approval: Da,2 //^ Date: NA Date: NA Date: ^ IV. Authorization to Resume Work W h ere halting o f w ork occurred, resum ption o f w ork m u st first b e app ro ved b y Lab orato ry M an a g e m e n t 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