Document gV3Oez186dv826r5gqp2XyM3

GLP10-01-02. Interim Report 35: Analysis of PFBS, PFHS, and PFOS in Groundwater Samples; Decatur Waste Water Treatment Plant (WWTP) Pump Test - June 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 h o n e :610-701-3761 Author Susan Wolf 3M Environmental Laboratory Interim Report Completion Date Date of Last Signature Performing Laboratory 3M Environmental Health and Safety Operations Environmental Laboratory 3M Center, Bldg 260-05-N-17 St. Paul, MN 55144 Project Identification GLP10-01-02-35 Total Number of Pages 74 The testing reported herein meet the requirements of ANSI/ISO/IEC 17025:2005 "General Requirements for the Competence of Testing and Calibration Laboratories", in accordance with the A2LA Testing Certificate # 2052.01. Testing that complies with this International Standard also meets principles of ISO 9001:2000. Testing Cert #2052.01 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 This page has been reserved for specific country requirements. Page 2 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 G LP C ompliance Statement , Report Title: GLP10-01-02, Interim Report 35: Analysis of PFBS, PFHS, and PFOS in Groundwater Samples; Decatur Waste Water Treatment Plant (VWVTP) Pump Test-June 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. Page 3 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Q uality A ssurance Statement Report Title: GLP10-01-02, Interim Report 35: Analysis of PFBS, PFHS, and PFOS in Groundwater Samples; Decatur Waste Water Treatment Plant (WWTP) Pump Test - June 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 08/09/12 Phase Data and Report Date Reported to Testing Facility Management Study Director 08/20/12 08/20/12 QTjtyi'l* Date Page 4 of 74 IA GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Ta b l e o f Co n t e n t s GLP Compliance Statement.......................................................................................................................3 Quality Assurance Statement.....................................................................................................................4 Table of Contents........................................................................................................................................5 List of T a b le s...............................................................................................................................................6 1 Study Information.................................................................................................................................7 2 Sum m ary..............................................................................................................................................8 3 Introduction...........................................................................................................................................8 4 Test & Control Substances.................................................................................................................9 5 Reference Substances......................................................................................................................10 6 Test S ystem ....................................................................................................................................... 11 7 Method S um m ary..............................................................................................................................11 7.1 M ethod...............................................................................................................................11 7.2 Sample Collection..............................................................................................................11 7.3 Sample Preparation...........................................................................................................11 7.4 Analysis..............................................................................................................................11 8 Analytical Results............................................................................................................................... 13 8.1 Calibration ..........................................................................................................................13 8.2 System Suitability ..............................................................................................................13 8.3 Limit of Quantitation (LO Q )...............................................................................................13 8.4 Continuing Calibration.......................................................................................................14 8.5 Blanks................................................................................................................................. 14 8.6 Lab Control Spikes (LC Ss)...............................................................................................14 8.7 Analytical Method Uncertainty..........................................................................................16 8.8 Field Matrix Spikes (FMS).................................................................................................16 Page 5 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 8.9 Lab Matrix Spikes (LMS)................................................................................................. 17 9 Data Summary and Discussion........................................................................................................17 10 Conclusion..........................................................................................................................................22 11 Data/Sample Retention.................................................................................................................... 22 12 Attachm ents.......................................................................................................................................22 13 Signatures..........................................................................................................................................23 Lis t o f Ta b l e s Table 1. Summarized PFBS, PFHS, and PFOS Results (WWTP Pump Test - June 2012).... 8 Table 2. Sample Description Key Code...................................................................................... 11 Table 3. Instrument Parameters.................................................................................................. 12 Table 4. Liquid Chromatography Conditions.............................................................................. 12 Table 5. Mass Transitions............................................................................................................ 13 Table 6 . Limit of Quantitation (LOQ)............................................................................................13 Table 7. Laboratory Control Spike Recovery..............................................................................15 Table 8. Analytical Uncertainty.................................................................................................... 16 Table 9. Field Matrix Spike...........................................................................................................17 Table 10. Laboraotry Matrix S p ike ..............................................................................................17 Table 11. DAL GW W W 8R 120627-1000.................................................................................. 18 Table 12. DAL GW W W 8R 120627-2130.................................................................................. 18 Table 13. DAL GW W W 8R 120628....................................................................................... 19 Table 14. DAL GW WW9R 120625....................................................................................... 19 Table 15. DAL GW WW9R 120625-2400.................................................................................. 20 Table 16. DAL GW WW9R 120626....................................................................................... 20 Table 17. DAL GW TRIP01 120625............................................................................................21 Page 6 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 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 Kevin Eich, Analyst Kelly Ukes, Analyst Study Dates Study Initiation: March 8, 2010 Interim 35 Experimental Termination: August 3, 2012 Interim Report Completion: Date of Interim Report Signing Location of Archives All original raw data and the analytical report have been archived at the 3M Environmental Laboratory according to 40 CFR Part 792. The test substance and analytical reference standard reserve samples are archived at the 3M Environmental Laboratory according to 40 CFR Part 792 Page 7 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 2 Summary The 3M Environmental Laboratory received groundwater samples from a pump test conducted at two monitoring wells at the 3M Decatur facility in Decatur, AL. A total of twenty sample bottles were received at the 3M Environmental Laboratory for perfluorooctane sulfonate (PFOS), perfluorohexane sulfonate (PFHS) and perfluorobutane sulfonate (PFBS), and included duplicate groundwater samples and a field matrix spike (FMS) sample from each sampling location. Samples also included one trip blank containing Milli-QTM water and a trip blank spike. All samples were logged into the laboratory information management system (LIMS) under project GLP10-01-02-35. The groundwater samples and trip blanks were received from Weston personnel on June 29, 2012. All of the samples were prepared and analyzed for PFBS, PFHS, and PFOS following 3M Environmental Laboratory Method ETS-8-044.1 "Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis". The average and relative percent difference (RPD) for the PFBS, PFHS, and PFOS concentrations detected in the primary and field duplicate ground water samples are summarized in Table 1. The trip blank sample was below the lower limit of quantitation (LLOQ) for all analytes, indicating adequate control of sample contamination during shipping and sample collections. The analytical method uncertainties associated with the reported results are: PFBS + 19%, PFHS + 17% and PFOS + 22%. Table 1. Summarized PFBS, PFHS, and PFOS Results (WWTP Pump Test - June 2012). Sampling Location DAL-GW-WW8R-120627-1000 DAL-GW-WW8R-120627-2130 DAL-GW-WW8R-120628 DAL-GW-WW9R-120625-1250 DAL-GW-WW9R-120625-2400 DAL-GW-WW9R-120626 DAL-GW-TRIP01 Blank PFBS Avg. Conc. (ng/mL) RPD 575 1.2% 489 3.3% 453 4.6% 233 5.2% 248 1.6% 254 3.1% <0.100 PFHS Avg. Conc. (ng/mL) RPD 2610 0.0% 2280 1.3% 2070 1.5% 1290 3.1% 1340 0.75% 1330 3.8% <0.100 PFOS Avg. Conc. (ng/mL) RPD 5280 0.38% 4550 0.44% 4170 0.48% 2620 3.8% 2610 0.38% 2550 9.8% <0.0928 (1) The analytical method uncertainties associated with the reported results are: PFBS 19%, PFHS 17%, and PFOS 22%. 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 a pump test conducted at two on-site monitoring wells at the 3M Decatur facility in Decatur, AL for PFOS, PFHS and PFBS in an effort to characterize regional groundwater conditions. Page 8 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 The 3M Environmental Laboratory prepared sample bottles (250 mL high-density polyethylene) which were shipped to Decatur, AL Weston personnel prior to field sampling. Sample bottle sets for each groundwater sampling location included a field sample, field sample duplicate, and a field spike sample. 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 isomer), PFHS (linear isomer), and PFOS (linear and branched isomers) prior to being sent to the field for sample collection. All sample bottles included the addition of 18O2-PFBS, 13C3-PFHS, and 13C8-PFOS (internal standard) at a nominal concentration of 1 ng/mL and 13C4-PFOS (surrogate recovery standard) at a nominal concentration of 0.1 ng/mL. Due to the level of PFBS, PFHS and PFOS detected in samples, and the need to dilute the samples 100-fold and 200-fold, the internal standards 18O2-PFBS, 13C3-PFHS, and 13C8-PFOS were not used for quantitation, nor could the surrogate recovery standard 13C4-PFOS be quantitated. In addition, the field matrix spike sample at 1000 ng/mL was not appropriate for all analytes at each sampling location. Where necessary, a laboratory matrix spike was prepared. See section 8.8 of the report for field matrix spike levels. See section 8.9 of the report for laboratory 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 LC/MS/MS; Direct Injection Analysis". Table 1 summarizes the average and relative percent difference (RPD) for the PFBS, PFHS, and PFOS concentrations detected in the primary and field duplicate ground water samples and the trip blank sample. Tables 10-15 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 9 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 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 (L in ear) Potassium Perfluorobutane sulfonate C 4F9S O 3K + NA Target Analyte Reference Standard 3M 01/10/2017 Frozen 41-2600-8442-5 TCR-121 White Powder 96.7% PFHS (L in ea r) Sodium Perfluorohexane sulfonate C 6F 13S O 3N a L-PFHXS Target Analyte Reference Standard W elling to n 03/25/2018 Frozen LPFHxSAM08 TCR08-0018 Crystalline 100% PFOS (L in e a r + B ranched) Potassium Perfluorooctane sulfonate C aF 17S O 3-K+ Br-PFOSK Target Analyte Reference Standard W elling to n 12/01/2014 Frozen brPFOSK1111 TCR11-0041 Liquid 99.9% R eference S ubstance Chemical Name Chemical Formula Identifier Use Source Expiration Date Storage Conditions Chemical Lot Number TCR Number Physical Description Purity PFOS (1) (L in e a r + B ranched) Potassium Perfluorooctane sulfonate C aF 17S O 3 K+ CAS # 2795-39-3 FMS Reference Standard S ig m a -A ld ric h 02/04/2014 A m b ie n t 1424328V TCR11-0028 White Powder 99.7% (1) Due to the level of PFOS detected in the samples, the FMS reference standard for PFOS was not utilized in this analysis. Page 10 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 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 Example D A L -G W -W W 9 R -D B -120625-2400 Sample Location DAL= Decatur, Alabama Sample Type G W = Ground Water Well ID Example: WW9R Sample Type 0=primary sample DB=duplicate sample FMS = field matrix spike Sampling Date 120635 = June 25, 2012 Sampling Time 2400 = 24:00 or 12:00 am Not all sampling locations included a sampling time in the sample description 7 Method Summary 7.1 Method 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 GLP10-01-02-35 were returned to the laboratory at ambient conditions on June 29, 2012. Samples were stored refrigerated at the laboratory after receipt. A set of laboratory prepared T rip Blank and a T rip Blank field matrix spike were sent with the set of sample collection bottles. 7.3 Sample Preparation Due to the level of PFBS, PFHS, and PFOS detected in all sampling locations, the primary and field duplicate samples were diluted 1:100 or 1:200. The 1:100 dilution was preapred by diluting 0.10 mL of a well mixed sample with 9.9 mL of Milli Q water. The 1:200 dilution was preapred by diluting 0.050 mL of a well mixed sample with 9.9 mL of Milli Q water (dilution factor of 199). The Trip Blank FMS sample was diluted 1:100 by diluting 0.10 mL of a well mixed sample with 9.9 mL of Milli Q water. The Trip Blank sample was aliquoted and analyzed with out sample dilution. 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 Page 11 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 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. Due to the nature of the sample, the wide range of concentrations found in the sample, and the environmental occurrence of multiple isomers of the laboratory's analytes of interest, the software used for processing the analytical results is not able to consistently integrate the analytical peak, manual integration of the analytical peak is necessary. All manual integrations are performed following the procedures outlined in method ETS-12-010. The consistency of the laboratory's integration is ensured through the training of laboratory personnel, the peer review process required for all manual integrations, the review of manual integrations by the QAU, and where necessary the review of manual integrations by laboratory management. 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 McCoy ETS-8-044.1 7/20/2012 and 8/3/2012 Agilent 1290 Betasil C18 (4.6 mm X 100 mm), 5|x Betasil C18 (4.6 mm X 100 mm), 5p 5 ixL Applied Biosystems API 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) 0.00 0.50 4.00 6.00 11.0 13.0 13.5 16.0 16.5 19.0 Flow Rate (fJmin) 750 750 750 750 750 750 750 750 750 750 Percent A (5 m M ammonium acetate:0.01% Acetic Acid) 90.0 90.0 70.0 70.0 20.0 20.0 10.0 10.0 90.0 90.0 Percent B (Acetonitrile) 10.0 10.0 30.0 30.0 80.0 80.0 90.0 90.0 10.0 10.0 Page 12 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Table 5. Mass Transitions. Analyte Mass Transition Q1/Q3 Reference Material Structure PFBS 299/80 299/99 Linear PFHS 399/80 399/99 Linear 499/80 PFOS 499/99 Linear + Branched 499/130 Dwell time was 30 msec for each transition. The individual transitions were summed to produce a "total ion chromatogram" (TIC), which was used for quantitation. 8 Analytical Results 8.1 Calibration Samples were analyzed against an external standard calibration curve. Calibration standards were prepared by spiking known amounts of the stock solutions containing PFBS, PFHS, and PFOS (reference standard containing both linear and branched isomers) into Milli Q water. Standards ranging from 0.10 ng/mL to 75 ng/mL (nominal) were analyzed (0.10 mg/mL to 25 ng/mL for PFBS analysis on 8/3/120) . 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. 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 PFBS, PFHS, and PFOS. The correlation coefficient (r) was greater than 0.995 for PFBS, PFHS, and PFOS. 8.2 System Suitability A calibration standard was analyzed four times at the beginning of each analytical sequence to demonstrate overall system suitability. The acceptance criteria of less than or equal to 5% relative standard deviation (RSD) for peak area and retention time criteria of less than or equal to 2% RSD was met for PFBS, PFHS and PFOS for each analysis. 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 7/20/12 7/20/12 7/20/12 8/3/12 NA = Not Applicable D ilu tio n F a c to r 1 100 199 100 PFBS LOQ, ng/m L 0.100 10.0 19.9 10.0 PFHS LOQ, ng/m L 0.100 10.0 19.9 NA PFOS LOQ, ng/m L 0.0928 9.28 18.5 NA Page 13 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 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 for each analysis. 8.5 Blanks Two types of blanks were prepared and analyzed with the samples: method procedural blanks and a trip blank. Method procedural blank results were reviewed and used to evaluate method performance and to determine the LOQ for PFBS, PFHS,and PFOS. The trip blank reflects the shipping and sample collection conditions the sample bottles and samples experience. 8.6 Lab Control Spikes (LCSs) Low, mid, and high lab control spikes were prepared and analyzed in triplicate. LCSs were prepared by spiking known amounts of the analytes into 10 mL of Milli Q water to produce the desired concentration. The method acceptance criteria, average of LCS at each level within 100% 20% with an RSD <20%, were met for PFBS, PFHS, and PFOS with the following exception. 7/20/12 Analysis: The low set of LCS samples for PFOS had an average recovery of 79.3%. A method deviation is included with the raw data. 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 * 1 0 0 % average LCS recovery Page 14 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Table 7. Laboratory Control Spike Recovery. ETS-8-044.1 Analyzed 7/20/12 Lab ID LCS-120720-1 LCS-120729-2 LCS-120729-3 Average %RSD LCS-120729-4 LCS-120729-5 LCS-120720-6 Average %RSD LCS-120720-7 LCS-120720-8 LCS-120720-9 Average %RSD S p ik e d C o n c e n tra tio n (ng/m L) PFBS C a lc u la te d C o n c e n tra tio n (ng/m L) % Recovery S p ik e d C o n c e n tra tio n (ng/m L) 0.200 0.200 0.200 0.199 0.196 0.210 101% 3.7% 99.6 97.9 105 0.200 0.200 0.200 2.00 2.00 2.00 1.89 1.85 1.94 94.6% 2.6% 94.3 92.4 97.2 2.00 2.00 2.00 20.0 20.0 20.0 18.4 19.5 19.7 95.9 3.5% 92.1 97.4 98.3 20.0 20.0 20.0 PFHS C a lc u la te d C o n c e n tra tio n (ng/m L) 0.189 0.162 0.188 89.9% 8.4% 1.95 1.96 1.90 96.9% 1.8% 18.8 20.3 19.8 98.1% 3.6% % Recovery 94.6 81.2 94.0 97.6 98.2 94.9 94.2 101 99.0 ETS-8-044.1 Analyzed 7/20/12 Lab ID LCS-120720-1 LCS-120729-2 LCS-120729-3 Average %RSD LCS-120729-4 LCS-120729-5 LCS-120720-6 Average %RSD LCS-120720-7 LCS-120720-8 LCS-120720-9 Average %RSD P FO S (Linear an d Branched) S p ik e d C o n c e n tra tio n (ng/m L) C a lc u la te d C o n c e n tra tio n (ng/m L) % Recovery 0.186 0.186 0.186 0.168 0.135 0.140 79.3% (1) 12% 90.5 72.5 75.0 1.86 1.86 1.86 1.74 1.66 1.60 89.7% 4.1% 93.3 89.4 86.0 18.6 18.6 18.6 17.4 18.6 17.4 95.6% 3.7% 93.4 99.7 93.6 (1) LCS did not meet acceptance criteria of 100 20%. Page 15 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Table 7 continued. Laboratory Control Spike Recovery. ETS-8-044.1 Analyzed 8/03/12 Lab ID LCS-120803-1 LCS-120803-2 LCS-120803-3 Average %RSD LCS-120803-4 LCS-120803-5 LCS-120803-6 Average %RSD LCS-120803-7 LCS-120803-8 LCS-120803-9 Average %RSD PFBS S p ik e d C o n c e n tra tio n (ng/m L) C a lc u la te d C o n c e n tra tio n (ng/m L) % Recovery 0.200 0.200 0.200 0.225 0.217 0.220 110% 1.8% 112 108 110 2.00 2.00 2.00 2.10 2.00 2.10 103% 2.8% 105 100 105 20.0 20.0 20.0 21.0 22.4 22.6 110% 4.0% 105 112 113 (1) LCS did not meet acceptance criteria of 100 20%. 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 expanded uncertainty is calculated by multiplying the standard deviation by a factor of 2 , which corresponds to a confidence level of 95%. Table 8. Analytical Uncertainty. Analyte PFBS PFHS PFOS Standard Deviation 9.39 8.77 10.8 Method Uncertainty 19% 17% 22% 8.8 Field Matrix Spikes (FMS) A field matrix spike was collected at each sampling point to verify that the analytical method is applicable to the collected matrix. Field matrix spikes were generated by adding a measured volume of field sample to a container spiked by the laboratory with PFBS (linear), PFHS (linear), and PFOS (linear 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. The field matrix spike sample was not appropriate for all analytes at each sampling location. Where necessary, a laboratory matrix spike was prepared. See section 8.9 of the report for laboratory matrix spike levels. Field matrix spike recoveries are presented in section 9 of this report. Page 16 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Table 9. Field Matrix Spike. Sampling Location All locations and Trip Blank PFBS, ng/mL 1002 PFHS, ng/mL 994 PFOS, ng/mL 1012 ( Sam ple Concentration o f FMS - Average C o nce ntration: Field Sam ple & Field Sam ple Dup.) FMS Recovery = --------- --------------------------------------------------------------------------------------------------------------------------------- * 100% Spike Concentraton 8.9 Lab Matrix Spikes (LMS) Due to the high level of PFHS and PFOS detected in several sampling locations, a laboratory matrix spike (LMS) sample was prepared to verify that the analytical method is applicable to the collected matrix. Laboratory matrix spikes were generated by adding a measured volume of the target analytes to an aliquot of the primary sample. The laboratory matrix spike added was based on the on-column instrument concentration. All sampling locations: the primary sample was diluted 1:200, to which a nominal concentration of 30 ng/mL was added for a LMS concentration of approximately 6000 ng/mL. Table 10. Laboraotry Matrix Spike Sampling Location All locations PFBS, ng/mL 6012 PFHS, ng/mL 6006 PFOS, ng/mL 5579 Lab 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. Lab matrix spike concentrations must be 50% of the sample concentration to be considered an appropriate field spike. Lab matrix spikes are presented in section 9 of this report. The following calculation was used to calculate the lab matrix spike recovery in Section 9 of the report: LMS Recovery = Sam ple Concentration of LMS - A verage C o nce ntration: Field Sam ple & Field Sam ple Dup.) * 1 0 0 % Spike Concentraton 9 Data Summary and Discussion The tables below summarize the sample results and field and/or laboratory matrix spike recoveries for the sampling locations as well as the Trip Blank. 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 and lab 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. All QC elements of the method; field matrix spikes (FMSs) and laboratory matrix spikes (LMSs) met method acceptance criteria. Page 17 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Table 11. DAL GW WW8R 120627-1000 PFBS PFHS PFOS 3 M L IM S ID D e s c rip tio n G LP 10-01-02-35-001 G L P 1 0 -0 1 -0 2 -3 5 -0 0 2 G L P 1 0 -0 1 -0 2 -3 5 -0 0 3 G LP10-01-02-35-001; LMS DAL-GW -W W 8R-0-120627-1000 DAL-GW -W W 8R-DB-120627-1000 DAL-GW -W W 8R-FMS-120627-1000 DAL-GW -W W 8R-0-120627-1000 A verag e Concentration (ng/m L) % R PD C o n c e n tra tio n (ng/m L) % Recovery 578 571 1560 6760 NA NA 98.4 103 57 5 n g/m L 1.2% C o n c e n tra tio n (ng/m L) % Recovery 2610 2610 3600 8800 NA NA NC 103 2610 ng/m L 0.0% C o n c e n tra tio n (ng/m L) % Recovery 5270 5290 5940 11300 NA NA NC 108 5280 ng/m L 0.38% NA = Not Applicable NC = Not Calculated; Spike level was less than 0.5x the endogenous sample concentration. Samples analyzed using a 1:199 dilution factor Table 12. DAL GW WW8R 120627-2130 PFBS PFHS PFOS 3 M L IM S ID D e s c rip tio n C o n c e n tra tio n C o n c e n tra tio n C o n c e n tra tio n (ng/m L) % Recovery (ng/m L) % Recovery (ng/m L) % Recovery G L P 1 0 -0 1 -0 2 -3 5 -0 0 4 G L P 1 0 -0 1 -0 2 -3 5 -0 0 5 G L P 1 0 -0 1 -0 2 -3 5 -0 0 6 GLP10-01 -02-35-004; LMS DAL-GW -W W 8R-0-120627-2130 DAL-GW -W W 8R-DB-120627-2130 DAL-GW -W W 8R-FMS-120627-2130 DAL-GW -W W 8R-0-120627-2130 497 481 1400 6500 NA NA 90.9 100 2290 2260 3170 8010 NA NA NC 95.5 4540 4560 5660 10400 NA NA NC 105 A verag e Concentration (ng/m L) % R PD 489 ng/m L 3.3% 2280 n g/m L 1.3% 4550 ng/m L 0.44% NA = Not Applicable NC = Not Calculated; Spike level was less than 0.5x the endogenous sample concentration. Samples analyzed using a 1:199 dilution factor Page 18 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Table 13. DAL GW WW8R 120628 3 M L IM S ID D e s c rip tio n G L P 1 0 -0 1 -0 2 -3 5 -0 0 7 G L P 1 0 -0 1 -0 2 -3 5 -0 0 8 G L P 1 0 -0 1 -0 2 -3 5 -0 0 9 GLP10-01-02-35-007; LMS DAL-GW -W W 8R-0-120628 DAL-GW -W W 8R-DB-120628 DAL-GW -W W 8R-FMS-120628 DAL-GW -W W 8R-0-120628 A verag e Concentration (ng/m L) % R PD PFBS PFHS PFOS C o n c e n tra tio n C o n c e n tra tio n (ng/m L) % Recovery (ng/m L) % Recovery 463 442 1400 6780 NA NA 94.6 105 2050 2080 3070 8010 NA NA NC 99.0 453 ng/m L 4.6% 2070 ng/m L 1.5% C o n c e n tra tio n (ng/m L) % Recovery 4160 4180 4490 10400 NA NA NC 112 4170 ng/m L 0.48% NA = Not Applicable NC = Not Calculated; Spike level was less than 0.5x the endogenous sample concentration. Samples analyzed using a 1:199 dilution factor Table 14. DAL GW WW9R 120625 PFBS PFHS PFOS 3 M L IM S ID D e s c rip tio n C o n c e n tra tio n C o n c e n tra tio n C o n c e n tra tio n (ng/m L) % Recovery (ng/m L) % Recovery (ng/m L) % Recovery GLP10-01 -02-35-010 GLP10-01 -02-35-011 GLP10-01 -02-35-012 GLP10-01-02-35-010; LMS DAL-GW -W W 9R-0-120625-1250 DAL-GW -W W 9R-DB-120625-1250 DAL-GW -W W 9R-FMS-120625-1250 DAL-GW -W W 9R-0-120625-1250 239 227 1170 6320 NA NA 93.5 101 1310 1270 2170 7150 NA NA 88.5 97.6 2570 2670 3290 8970 NA NA NC 114 A verag e Concentration (ng/m L) % R PD 233ng/m L 5.2% 1290 ng/m L 3.1% 2620 ng/m L 3.8% NA = Not Applicable NC = Not Calculated; Spike level was less than 0.5x the endogenous sample concentration. Samples analyzed using a 1:199 dilution factor Page 19 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Table 15. DAL GW WW9R 120625-2400 PFBS PFHS PFOS 3 M L IM S ID D e s c rip tio n C o n c e n tra tio n C o n c e n tra tio n C o n c e n tra tio n (ng/m L) % Recovery (ng/m L) % Recovery (ng/m L) % Recovery GLP10-01 -02-35-013 GLP10-01 -02-35-014 GLP10-01 -02-35-015 GLP10-01-02-35-013; LMS DAL-GW -W W 9R-0-120625-2400 DAL-GW -W W 9R-DB-120625-2400 DAL-GW -W W 9R-FMS-120625-2400 DAL-GW -W W 9R-0-120625-2400 250 246 1210 N A (1) NA NA 96.0 N A (1) 1330 1340 2590 7110 NA NA 126 96.2 2600 2610 3710 7970 NA NA NC 96.2 A verag e Concentration (ng/m L) % R PD 248 ng/m L 1.6% 1340 ng/m L 0.75% 2610 ng/m L 0.38% NA = Not Applicable NC = Not Calculated; Spike level was less than 0.5x the endogenous sample concentration. PFHS and PFOS diluted 1:199 and reported from 7/20/12 analysis. PFBS diluted 1:100 and reported from 8/3/12 analysis. (1) Sample was not analyzed. Table 16. DAL GW WW9R 120626 3 M L IM S ID D e s c rip tio n GLP10-01 -02-35-016 GLP10-01 -02-35-017 GLP10-01 -02-35-018 GLP10-01-01 -36-016; LMS DAL-GW -W W 9R-0-120626 DAL-GW -W W 9R-DB-120626 DAL-GW -W W 9R-FMS-120626 DAL-GW -W W 9R-0-120626 A verag e Concentration (ng/m L) % R PD PFBS PFHS PFOS C o n c e n tra tio n C o n c e n tra tio n C o n c e n tra tio n (ng/m L) % Recovery (ng/m L) % Recovery (ng/m L) % Recovery 258 250 1140 6050 NA NA 88.4 96.4 1350 1300 2180 7130 NA NA 86.0 96.7 2670 2420 3180 8460 NA NA NC 106 254 ng/m L 3.1% 1330 ng/m L 3.8% 2550 ng/m L 9.8% NA = Not Applicable NC = Not Calculated; Spike level was less than 0.5x the endogenous sample concentration. Samples analyzed using a 1:199 dilution factor Page 20 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Table 17. DAL GW TRIP01 120625 3 M L IM S ID GLP10-01 -02-35-019 G L P 1 0 -0 1 -0 2 -3 5 -0 2 0 D e s c rip tio n DAL-GW -TRIP01-0-120625 DAL-GW -TRIP01-FM S-120625 NA = Not Applicable FMS Sample analyzed using a 1:100 dilution factor PFBS PFHS PFOS C o n c e n tra tio n C o n c e n tra tio n (ng/m L) % Recovery (ng/m L) <0.100 947 NA 94.5 <0.100 939 % Recovery NA 94.5 C o n c e n tra tio n (ng/m L) <0.0928 785 % Recovery NA 77.6 Page 21 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 10 Conclusion Laboratory control spikes, field matrix spikes, and lab 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 35 (General Project Outline) Attachment B: Representative Chromatograms and Calibration Curves Attachment C: Analytical Method-ETS-8-044.1 Attachmetn D: Protocl Amendment GLP10-01-02-35 Deviation Page 22 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 13 Signatures Cleston Lange, Ph.D., 3M Principal Analytical Investigator f / zif/. Date William K. Reagen, Ph.D., 3M Environmental Laboratory Technical Director Date Page 23 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 At t a c h m e n t A: Pr o t o c o l A m e n d m e n t Page 24 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Analytical Protocol: GLP10-01-02 Amendment 35 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, 35 Amendment Date: July 11, 2012 Performing Laboratory 3M Environmental, Health, and Safety Operations 3M Environmental Laboratory Building 260-5N-17 Maplewood, MN 55144-1000 Laboratory Project Identification GLP10-Q1-02 Sampling Event 3M Decatur WWTP Area Pump Test Page 1 of 6 Page 25 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Analytical Protocol: GLP10~Q1~Q2 Amendment 35 This amendment modifies the following portion of protocol: "Analysis of PFOS, PFHS and PFBS in Groundwater, Soil and Sediment for the 3M Decatur Phase 3 Site-Related Monitoring Program" Protocol reads: No changes to the wording of the protocol are required. A m ubd to read : No changes to the wording of the protocol are required. This amendment only addresses and documents the addition of the General Project Outiine (GPO) for the collection and analysis of groundwater samples as part of the 3M Decatur Phase 3 Program for PFOS, PFHS and PFBS (GLP1 0-01-02), The anticipated sample collection will occur around the timeframe of the week of June 25, 2012 The groundwater samples for this sampling event will be entered into the 3M Environmental Laboratory LIMS as project GLP10-01-02-35 and reported as interim report GLP10-01-02-35, (reflecting study GLP10-01-02 and amendment -35). Reason; The reason for this amendment is to document the General Project Outline (GPO) which describes the anticipate groundwater sample collection event for two new monitoring wells in the vicinity of the Wastewater Treatment Plant (WWTP) at the 3M Decatur facility. The GPO is three pages in length and included as attached to this amendment form. Page 2 of 6 Page 26 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Analytical Protocol: GLP10-01-02 Amendment 35 Amendment Approva! / / j uL y Page 3 of 6 Page 27 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Analytical Protocol: GLP10-01-02 Amendment 35 Environmental Health & Safety Operations, Environmental Laboratory General Project Outline To: Gary Hohenstein, 3M EHS&Opns From: Susan Wolf, 3M EHS&Opns, Environmental Lab cc: William Reagen, 3M EHS&Opns; Environmental Lab Cleston Lange, 3M EHS&Opns; Environmental Lab Jai Kesari, Weston Solutions Date: July 10, 2012 Subject: Analysis of Periluorooctane Sulfonate (PFOS), Perfluorohexane Sulfonate (PFHS) and Periluorobutane sulfonate (PFBS) in Groundwater, Soil and Sediment for the 3M Decatur Phase 3 Site-Related Monitoring Program; GLP Interim Report 35; 3M Decatur Wastewater Treatment Plant (WWTP) Area Pump Test - June 2012 1 General Project Information C ontacts Lab Request Num ber S ix D igit D ep artm ent N um ber P ro ject S chedu ie/Test Dates 3 Ni S p o n s o r R e p re s e n ta tiv e Gary Hohenstein 3M EHS Operations 3M Building 224-5W-03 Saint Paul, MN 55144-1000 Phone: (651)737-3570 aahohensteiniEernmm.com 3M Environm ental Laboratory M anagem ent William K. Reagen 3M EHS Opns, Environmental Laboratory 260-5N-17 651 733-9739 wkreaqenfEDmmm.com Principal A nalytical In vestigator Cleston Lange 3M EHS Opns, Environmental Laboratory 260-5N-17 651 733-9860 cclanaefi5mmm.com S am pling C oordinator Timothy Frinak Weston Solutions Timothv frinakOwestonsolutions com Phone: (334)-332-9123 G LP10-01-02-35 Dept #530711, Project #0022674449 Sampling scheduled for the week of June 25, 2012 All verbal and written correspondence will be directed to Gary Hohenstein. Page 4 of 6 Page 28 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Analytical Protocol: GLP10-01-02 Amendment 35 2 Background Information and Project Objective(s) The 3M EHS Operations Laboratory (3M Environmental Lab) will receive and analyze groundwater samples collected from two new wells located in the vicinity of the 3M Decatur wastewater treatment plant (WWTP) for Perfluorobutanesulfonate (PFBS), Perfiuorohexanesutfonate (PFHS), and Perfluorooctanesulfonate (PFOS), Analyses will be conducted under the GLP requirements of EPA TSCA Good Laboratory Practice Standards 40 CFR 792. Groundwater samples will be collected by Weston Solutions personnel the week of June 25, 2012. The 3M Environmental laboratory will prepare the sample bottles with all required spikes to ensure that resuits for PFBS, PFHS, and PFOS are of a known precision and accuracy. The final report will be submitted to Gary Hohenstein and Jai Kesari upon completion under interim report GLP10-01-02-35. 3 Project Schedule Sample collection bottles will be prepared by 3M Environmental Laboratory for sampling the week of June 25, 2012. Sample bottles were shipped in coolers overnight to 3M Decatur for arrival on Friday, June 22, 2012. Martin Smith \ Weston Trailer 3M Decatur Plant 1400 State Docks Road Decatur, Alabama 35601 4 Test Parameters The targeted limit of quantitation will be 0.025 ng/mL (ppb) for PFBS, PFHS, and PFOS. Two sampling locations have been specified; WW8R and WW9R. These are new wells with no historical sampling values for estimating field matrix spike levels, however, information received from Weston indicates that the new welts are located between wells TW2R and TW3R sampled in April 2012 and are expected to have PFBS, PFHS, and PFOS concentrations similar to these locations. Given this information, at each sampling location, a total of three sample bottles will be collected (sample, sample duplicate and high field matrix spike). The field matrix spike will be prepared at 1000 ng/mL. The "fill to here" line on each 250 mL Nalgene bottle will be 200 m l. One set of trip blanks consisting of reagent-grade water as well as a trip blank spike will be prepared at the 3M Environmental Laboratory and sent to the sampling location with the other bottles. While the samples are expected to have high levels of PFBS, PFHS, and PFOS, all sample bottles will include the addition of 130 2-PFBS, ieC>2-PFHS, and 13Cs-PFOS (internal standard) at a nominal concentration of 1 ng/mL and 13C4-PFOS (surrogate spike) at a nominal concentration of 0.1 ng/mL. If the sample concentrations are as high as expected and sample dilution is necessary, the use of the internal standards and reporting of the surrogate recovery standard will not be possible since both will be diluted out of reporting range. If the field matrix spike level is not appropriate as compared to the sample concentration, a laboratory matrix spike may be prepared to access accuracy. Page 5 of 6 Page 29 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Analytical Protocol: GLP10-01-02 Amendment 35 5 Test Methods Samples will be prepared and analyzed by LC/MS/MS following ETS-8-044.1 "Method of Analysis for the Determination of Perfluorinated Compounds In Water by LC/MS/MS; Direct Injection Analysis". The data quality objectives for these studies are quantitative resuits for the target analytes with an analytical accuracy of 10Q30%. Field matrix spikes not yielding recoveries within 10030% will be addressed in the report and the final accuracy statement may be adjusted accordingly. Where applicable, samples will be analyzed against an internal standard calibration curve. Each curve point will contain isotopically-labeled perfluorocarboxylic acids and perfluorosuffonic acids at a nominal concentration of 1 ng/mL. The calibration curve will be generated by taking the ratio of the standard peak area counts over the internal standard peak area counts to fit the data for each analyte. 6 Reporting Requirements For each sampling location, the report will contain the results for the sample, sample duplicate, and field matrix spike. Trip blank and trip blank spike will be reported for the sampling event. Laboratory control spikes of reagent water prepared at the time of sample extraction will also be reported and used to evaluate the overall method accuracy and precision Method blanks of reagent water prepared at the time of sample extraction will be used to determine the method detection limit. Page 6 of 6 Page 30 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 At t a c h m e n t B: Re p r e s e n t a t iv e Sa m p l e Ch r o m a t o g r a m s a n d Ca l ib r a t io n Cu r v e (s ) Page 31 of 74 *ETS-McCoy GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Results Name: mc120720a.rdb Printing Time: 8:38:39 AM Page 32 of 74 *ETS-McCoy GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Results Name: mc120720a.rdb Printing Time: 8:28:09 AM Page 33 of 74 *ETS-McCoy GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Results Name: mc120720a.rdb Printing Time: 8:28:32 AM Page 34 of 74 *ETS-McCoy GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Results Name: mc120720a.rdb Printing Date: Wednesday, August 08, 2012 Data printed by STW Printing Time: 8:43:56 AM Page 1 of 12 Page 35 of 74 *ETS-McCoy GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Results Name: mc120720a.rdb Printing Date: Wednesday, August 08, 2012 Data printed by STW Printing Time: 8:43:56 AM Page 2 of 12 Page 36 of 74 *ETS-McCoy I PSaemakplNeaNmaem: e":PF"BmSc"120M72a0sas0(e5s0)": "2S9a9m.0p0l0e/8ID0.:0"0G0LDPa1.02-9091.-00020-3/959-0.01030" DaF"ile: "mc120720a.wiff S a mC opmlem eInntd: e"Dx A: L-GW -W5W1 9R-0-120625-2400 (1:200 dil)" Annotation: "" AACCS accoamqqln..cpculeDTelnaiamttTtreeeyad::pt ieoC: no:n c: 37U:/20nk16nN2/:2o03/w1A7! n ng/m M odified: Yes NP rooisce. AP el gr coer ni tthamg e: : S p e c i5fi; BRPeeap>saek. MM iinn Smo' - SSpu lbi.t :: 1 . WFinadcotwo HW eidigthh:t : W idth: :r : 21 3 0 .0 Us cRtee lda t iRvT 1 0 .0 In t. Type R etention AH reeiag h: t : End Tim Ma 849.50502e+ 0 0 c4 9 .7 0 10 . 0 tpss 9.98 8.0 Sam ple N am e: "m c120720a057" Sam ple ID: "GLP10-01-02-35-018" File: "mc120720a.wiff mPCeopamlekmNeIannmtd: ee":Dx"AP: LFB-GSW" -WM5W8as9sR(e-Fs)M: S"2-19290.062060/(810:.200000 dDil)a",29A9.n0n0o0ta/9ti9o.n0:00"" Da" CS aomnpcleen tTrya pt ieo: n : AAC ccaqql ..c uDTl aiamtteeed:: C onc U n k nNo/Aw n 57 :/221211/:4200.1 2AM ng/m L 2.1e5 2.0e5 MNP roooidscei.f iAePdel:gr coer inthtamg B a se . S ub. Wind sp Yesf y MMR eiinnp o.. r HW eidigthh:t : W idth: EU xsep eRc teelda t iRvT T : 3 0 .0 N1o0 .0 RI nett.e nTt iyopne A rea: H eight: End Tim : M 9a.n9u7a l 431768 co 2 .1 5 e + 0 0 5 91 .06.91 ts ps Time, min Printing Date: Wednesday, August 08, 2012 Data printed by STW Printing Time: 8:43:56 AM GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Results Name: mc120720a.rdb IPSaemakplNeaNmaem: e":PF"BmSc"120M72a0sas0(e5s5)": "2S9a9m.0p0l0e/8ID0.:0"0G0LDPa1,02-9091.-00020-3/959-0.01060" DaF"il a mCopmlem eInntd: "DAL-GW -W W9R-0-120626 (1:200 dil)" Annotation: C o npcl ee n tTrya pt ieo: n N/A AAC ccaqql ..c uTDl aiamtteeed:: Co 74 /:24 132/5:24801.1 2AM g/m L M odified: NP rooics e. AP el gr coerni tthamg e: : : BP ea saek. - SSpulbi.t . WFinadcotwo R e p o r t L a r g e s t Pe< MM iinn .. PP ee aa kk HW eidigthh:t : RSTm oWoitnhdionwg : W id th : EU xsep eRc teelda t iRvT :i f5y0 P a r, 1 .0 0 In t. Type R etention AH reeiag:h t : End Tim 8000.0 6000.0 4000.0 2000.0 0.0 ISam ple N am e: "m c120720a058" Sam ple ID: "GLP10-01 -01-36-016; LMS' a mPCeopamlekmNeIannmtd: e"e:Dx"AP: LFB-GSW" -WM5W9as9sR(e-0s-)1: 2"0269296.0(010:2/8000.0d0il0)" D aA,2n9n9o.t0at0i0on/9:9"."000 Da Caomncpelen tTr aytpieo: n: U nknNo/wAn AACccaqqlc.. u DTl aiamtteeed:: Co n c : 57 :/426120/5:21006.1 2AM ng/m L 1 00e6 9.50e5 MNP rooodicsi ef. i eAPdel rg:coerni thm : S p eYcei5sf 0y Pa r a m e te r s - MQ I I I BP eaaske MMR eiinnpo.. .S r t htPPCeei ub naaakkgr . Win d o w gHWWee,i st di gt hh P t: e : ra: :k : 21 .0 0 Yes 00 ..00 00 RT Win d o w : 3 0 .0 m in cps ec p o in ts Use R e la tiv e RT: N1 o0 .0 9.00e5 8.50e5 8.00e5 7.50e5 7.00e5 6.50e5 RI ne tte. n Tt iyopne : T i A rea H eight: S ta rt Time: M a9n.u9a7l 222 8465 cou ts 1 .05e+006 ps 91 .06.81 6.00e5 CO 5.50e5 I 5.00e5 4.50e5 4.00e5 3.50e5 3.00e5 2.50e5 2.00e5 1 50e5 1 00e5 5.00e4 0.00 Page 3 of 12 Page 37 of 74 *ETS-McCoy GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Results Name: mc120720a.rdb Printing Date: Wednesday, August 08, 2012 Data printed by STW Printing Time: 8:43:56 AM Page 4 of 12 Page 38 of 74 *ETS-McCoy GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Results Name: mc120720a.rdb Printing Date: Wednesday, August 08, 2012 Data printed by STW Printing Time: 8:43:56 AM Page 5 of 12 Page 39 of 74 *ETS-McCoy GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Results Name: mc120720a.rdb Printing Date: Wednesday, August 08, 2012 Data printed by STW Printing Time: 8:43:57 AM Page 6 of 12 Page 40 of 74 *ETS-McCoy GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Results Name: mc120720a.rdb Printing Date: Wednesday, August 08, 2012 Data printed by STW Printing Time: 8:43:57 AM Page 7 of 12 Page 41 of 74 *ETS-McCoy GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Results Name: mc120720a.rdb Printing Date: Wednesday, August 08, 2012 Data printed by STW Printing Time: 8:43:57 AM Page 8 of 12 Page 42 of 74 *ETS-McCoy GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Results Name: mc120720a.rdb Printing Date: Wednesday, August 08, 2012 Data printed by STW Printing Time: 8:43:57 AM Page 9 of 12 Page 43 of 74 *ETS-McCoy GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Results Name: mc120720a.rdb Printing Date: Wednesday, August 08, 2012 Data printed by STW Printing Time: 8:43:57 AM Page 10 of 12 Page 44 of 74 *ETS-McCoy GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Results Name: mc120720a.rdb Printing Date: Wednesday, August 08, 2012 Data printed by STW Printing Time: 8:43:57 AM Page 11 of 12 Page 45 of 74 *ETS-McCoy I PSaemakplNeaNmaem: e":PF"mOSc1" 207M2a0sas0(7e2s)": "4S9a9m.0p0le0/8ID0:.0"0L0CDS-a1.2409791.090-10"/99.F0i0le0: "Dmac.4192907.02000a/.w13if0f."000 Da" S a mC opmlem eInntd: e"0x.:2ppb LCS"73 Annotation: "" CS aomnpclee n tTryapt ieo: n : 0 .1 8Q6C ng/m L AACccaqql .c. uDTl aiamtteeed:: C o n c: 1 07 /:21015.1/:2460481 2AM ng/m L M odified: Yes NP rooics e. AP el gr coerni tthamg e:: sp BP ea saek. - SSpulbi.t . WFinadcotwo R e p o rt L a rg e s t Pe. MM iinn .. PP ee aa kk HW eidigthh:t : RSTm oWoitnhdionwg : W id th : 3U.L EU sxep eRc teelda t iRvT In t. Type R etention AH reeiag h: t : Ma 3 7. 7U3U5e+U Uc3 tpss End Tim 12.22 Time, min GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Results Name: mc120720a.rdb Printing Date: Wednesday, August 08, 2012 Data printed by STW Printing Time: 8:43:57 AM Page 12 of 12 Page 46 of 74 *ETS McCoy GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Results Name: mc120803a.rdb Printing Time: 9:10:14 AM Printing Date: Monday, August 06, 2012 *Data printed by KJU Page 1 of 1 Page 47 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Pr in t i n g Date: We dn es da y, A u g u s t 08, 2012 Data printed by STW Page 1 of 2 Page 48 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Pr in t i n g Date: We dn es da y, A u g u s t 08, 2012 Data printed by STW Page 2 of 2 Page 49 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 At t a c h m e n t C: A n a l y t ic a l Me t h o d (s ) Page 50 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 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 51 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 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 Num ber (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 a1nTdheCm8 eptehroflduoisrosaulpkpaonretesdulbfyonvaamlididaetiionnlwabiothraitnotreyrncaolnsttraonldsaarmdpclaelsiburnadtieorn3fMormC4et-hCo1d3vPaFliCdAatsio, nCE4,1C1-60,6a6n7d. C8 PFSAs, MM2(PGeeauttrhhtidooAadd,nssVceoecaftlfiAioodrnnaeat5isloy)tnasob"ifsl,iDbisnh.i)riSenEcugPtpiAmvpeoeMrt9the1oot/dfh4Po1Qdr4eC,5-Sr3CeA7gr,iNitsaeCtnrridaOatic/bo3.a)0nsE2De9uda/r9tooa9npReraeae.vn)q.FuC4iDor(eAm1m1mMe/0insa7tsys/i0of20no0):r.0AG1,nun"idGeaxuniIcdIea(nfPocarertGfAoer,nIesnredactutiisnotgrnya4,n)BdaionRadenpAaolnrynttiiencxgalIII 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 52 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 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 53 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 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 54 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 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 55 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 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 56 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 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- C 4]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 57 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 7.4 Stock Standard Solution (SSS) and Working Standard Solution Preparation The following standard preparation procedure serves as an example. Weighed amounts and final volumes may be changed to suit the needs of a particular study. For example, pL volumes may be spiked into volumetric flasks when diluting stock solutions to appropriate levels. 100 pg/mL target analyte SSSs--Weigh out 10 mg of analytical standard (corrected fo r p e rce n t salt, a c id [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 3PFAOrAep/oPrFtOsuSmcmanarbiezifnogunand ainss3eMssmreepnotrotfEt1he1-u0s5e60o.freference standards containing certified linear and branched isomers of 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 58 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 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-Cq]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 59 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Table 1. Stable Isotope PFCAs and PFSAs used for ISs and SRSs Coimpound Name 13C4-Perfluorobutanoic acid 13C4-Perfluoropentanoic acid 13C2-Perfluorohexanoic acid 13C4-Perfluoroheptanoic acid 13C8-Perfluorooctanoic acid 13C9-Perfluorononanoic acid 13C6-Perfluorodecanoic acid 13C7-Perfluoroundecanoic acid 13C2-Perfluorododecanoic acid 18O2-Ammonium Perfluorobutane sulfonate 13C3-Ammonium Perfluorohexane sulfonate 13C8-Sodium Perfluorooctane sulfonate 13C8-Perfluorooctanesulfonamide 13C4-Perfluorooctanoic acid 13C2-Perfluoroundecanoic acid 13C8-Perfluorooctane sulfonate Synonym or Acronym [1,2,3,4-13C4]PFBA [1,2,3,4,5-13C5]PFPeA [1,2 -13C2]PFHxA [1,2,3,4-13C4]PFHpA [1,2,3,4,5,6,7,8-13C8]PFOA [1,2,3,4,5,6,7,8,9-13C9]PFNA [1,2,3,4,5,6 -13C6]PFDA [1,2,3,4,5,6,7 -13C7]PFUnA [1,2 -13C2]PFDoA [18O2]PFBS [1,2,3-13C3]PFHS [1,2,3,4,5,6,7,8-13C8]PFOS [1,2,3,4,5,6,7,8-13C8]FOSA [1,2,3,4-13C4]PFOA [1,2 -13C2]PFUnA [1,2,3,4-13C4]PFOS Analytical Purpose IS for PFBA IS for PFPeA IS for PFHxA IS for PFHpA I13SCf4o]rPPFOFOAA and [1,2,3,4 IS for PFNA IS for PFDA IS for PFUnA IS for PFDoA, *PFTA IS for PFBS IS for PFHS IPSFOfoSr[P1F,2O,3S,4an13dC4], IS for FOSA SRS for all PFCAs: C4-C8 RSoeuferrceence Standard 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) R(ITndIiIvnidteuranla)tional Wellington Labs (Mix or Individual) Wellington Labs (Mix or Individual) Wellington Labs (mix) R(ITndIiIvnidteuranla)tional Wellington SRS for all PFCAs C9-C13 Wellington CSR6,SafnodrCal8l PFSAs: C4, 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 60 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Table 2. Example Preparation of Calibration Curve with ISs and SRSs Sam ple Description 0.025 ng/mL curve point 0.030 ng/mL curve point 0.04 ng/mL curve point 0.05 ng/mL curve point 0.1 ng/mL curve point 0.25 ng/mL curve point 0.5 ng/mL curve point 1 ng/mL curve point 2.5 ng/mL curve point 5.0 ng/mL curve point 10.0 ng/mL curve point 25.0 ng/mL curve point 50.0 ng/mL curve point 75.0 ng/mL curve point 100 ng/mL curve point Concentration o f WS, pg/mL 0.10 0.10 0.10 0.10 0.10 0.10 1.0 1.0 10.0 10.0 10.0 10.0 10.0 10.0 10.0 Volume of WS, pL 25 30 40 50 100 250 50 100 25 50 100 250 500 750 1000 Volume o f IS (2 pg/m L), pL 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 Concentration o f 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 A S TM Type I Water, or o ther su itable s o lv e n t(1>, m L 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 61 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 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 62 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 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 63 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 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 64 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 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 W ater 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 65 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 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 (m in ) 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 (M eth an o l) 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 66 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Table 4 Suggested MRM Transitions for Target Analytes, Surrogates, and Internal Standards Analyte PFBA (C4 Acid) PFPeA (C5 Acid) PFHxA (C6 Acid) PFHpA (C7 Acid) PFOA (C8 Acid) PFNA (C9 Acid) PFDA (C10 Acid) PFUnA (C11 Acid) PFDoA (C12 Acid) PFTA (C13 Acid) FBSA (C4 Sulfonamide) FOSA (C8 Sulfonamide) PFBS (C4 Sulfonate) PFHS (C6 Sulfonate) PFOS (C8 Sulfonate) [1,2,3,4 -13C4]PFB A [1,2,3,4,5 -13C 5]PFPeA H ,2 -13C 2lP F H xA [1,2,3,4- 13C4lP F H pA [1,2,3,4,5,6,7,8-13C 8lPFO A H ,2,3,4,5,6,7,8,9-13C 9lPFN A r 1,2,3,4,5,6 -13C 6lPFD A r 1,2,3,4,5,6,7 -13C 7lP F U nA [1,2 -13C 2lP F D oA [18O 2lPFB S [1,2,3-13C 3lPFH S [1,2,3,4- 13C4lPFO S [ 1,2,3,4,5,6,7,8-13C 8lFO SA [1,2,3,4-13C4lPFO A [1,2,3,4- ^ lP F O S [1,2 -13C 2lP F U nA Analyte Description Target Target Target Target Target Target Target Target Target Target Target Target Target Target Target IS for PFBA IS for PFPeA IS for PFHxA IS for PFHpA IS for PFOA IS for PFNA IS for PFDA IS for PFUnA IS for PFDoA and PFTA IS for PFBS IS for PFHS IS for PFOS IS for FOSA Surrogate (C4-C8 Acids) Surrogate(Sulfonates, FOSA) Surrogate (C9-C13 Acids) Mass Transition 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 Mass Transition Q3 (amu) 169 219 269, 119 319, 169 369, 219, 169 419, 169, 219 469, 269, 219 519, 269, 219 569, 169, 319 619, 369, 319 78 78 99, 80 99, 80 80, 99, 130 172 223 270 322 376 427 474 525 570 84 80 80 80 372 80 520 Multiple transitions for monitoring the analytes is an option. The use of one daughter ion is acceptable if data sensitivity and selectivity is achieved and provided that retention time criteria are met to assure adequate specificity. While the daughter ions may be chosen at the discretion of the analyst, mass transition 99 is suggested for PFOS. Quantitation may be performed using the total ion chromatogram (TIC, or summed MRMs) for a given analyte. For example, the PFOA TIC would sum all three of the monitored transitions. Use of the suggested primary ion is recommended. Retention times may vary slightly, on a day-to-day basis, depending on the batch of mobile phase and the gradient, column, guard column(s) used etc. Drift in retention times is acceptable within an analytical run, as long as the drift continues through the entire analysis and the standards are interspersed throughout the analytical run. 11.2 Calibration Curve Quantitation is by internal standard or external standard calibration. Calibration standards may be prepared in ASTM Type I, HPLC water, other suitable water, or a solvent/water mixture. If internal standard calibration does not meet calibration acceptance criteria, external calibration can be applied. See Table 1 for ETS-8-044.1 Page 17 of 22 Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis Page 67 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 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. 43M Environmental Laboratory study E08-0096 evaluated the effect on these results as a function ofthe number ofMRMs 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 68 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 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 Concentration (----) - Concentration of Sample (----) mL mL ng Spike Concentration (----) 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) ^ 100 Analyte added (ng/mL) 13 Analysis Batch Method Performance Criteria Any method performance parameters that are not achieved must be considered in the evaluation of the data. Nonconformance to any specified parameters must be described and discussed in the final report if the Technical Manager (non-GLP study) or Study Director (GLP study) chooses to report the data. If criteria listed in this method performance section are not met, maintenance may be performed on the system and samples reanalyzed, or other actions taken as appropriate. Document all actions in the raw data. If data are to be reported when performance criteria have not been met, the data must be footnoted on tables and discussed in the text of the report. 13.1 System Suitability - Analysis Batch A minimum of three system suitability samples should be injected at the beginning of each analytical run. These samples are run prior to the calibration curve. It is suggested that the system suitability injections have area counts with a target RSD of <5% and a target retention time RSD of <2%. There is no defined acceptability limit on these results as the %RSDs are dependent on the number of MRM transitions being monitored in the LC/MS/MS run or time period. Any effect of system suitability is incorporated in the QC acceptance criteria. 13.2 Calibration and Limit of Quantitation (LOQ) - Analysis Batch 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 69 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 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 5Method uncertainty based on INTERNATIONAL ANS/ISO/IED STANDARD 17025 reference (GUM, Guide to the Expression of Ub"Q.n)Gucaeenrottarigfinyiatiynng,inTUhMnocemearastsua,rien"mEtysetininmt)A.antMiaolneyttohicfoaLdlaaMbpoepralaisctuoarrteyiomAnendnaet,lm"ytoSicneascltorUnatdnecdEedirnittaiEionTnt;ySE-U1d2siit-no0gr1s2:L,aScb.iLoti.rnRagt.orrEeyflelCirseoonnnct,erMso:l.aS.Ra)mEospUsllReesAi,n"C,EaHnnEvdiMrAo/.nCmWITeiAnlltiCaalmGTseu.sidtien,g & 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 70 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 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. AEMvneaaallsuyuasrtieisnm,gNeaonnvtdUemEnxcbeperrrt/eaDsisneitcnyegminthbTeereUs2tn0icn0eg0r".ta,ciJ.nu)tTlyyaoy2lf0oN0r2,I.SBT.NM. eaansdurCeEm.enKtuRyeasttu,lNtsI."SdT.)TAedcahmnsic,aTl.NMo.,te"A1229L7A, 1G9u9i4deEdfoitriothne: "EGstuimidaetliionnesoffor 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 71 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 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 o f internal standard calibration by this method. Section 2. Included the use o f internal standard calibration by this method. Included the use o f 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 instrum ent parameters to section 10 o f the method. Section 7. Updated reference standards to include internal standards and surrogates. Changed concentration levels for working standards and included the use o f 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 o f LCSs. Included the use o f m ethanol 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 M ethod 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 o f analytical method uncertainty and quantitation o f 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 72 of 74 At t a c h m e n t D: De v ia t io n (s ) GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS in Groundwater Samples Decatur WWTP Pump Test - June 2012 Page 73 of 74 GLP10-01-02; Interim Report 35 Analysis of PFBS, PFHS, and PFOS In Groundwater Samples Decatur WWTP Pump Test - June 2012 Record of D eviation/N o nconform ance 1. Identification Study / Project No. Date(s) of Occurrence: Document Number: ETS-8-044.1 GLP10-01-02-35 7/20/12 Deviation type 0 SOP Equipment Procedure Method (Check one) Protocol GPO Other: __________________ II. Description (attach extra pages as needed) Method Requirements: 1. LCS: average recovery within 20% (section 13.4). Actual procedure/process: 1. The lo w se t of LCS sam ples for PFOS had an average recovery o f 79.3% III. Actions Taken ___________________________ (such as amendment issued, SOP revision, etc.)__________________________ Corrective Action ( Yes 0 No) Reference: Acceptability of the nonconforming work: 1. All LC Ss were used to determ ine the overall m ethod uncertainty. Since the LCS bias w as less than the overall method uncertainty determined by ETS-12-012.2 (22%) the method uncertainty was not expanded due to the recovery of the batch QC. A ctions: H alting o f W o rk C lient N otification W o rk R ecall 0 O th e r: Deviation will be noted in section 9 of the final report. W ithholding o f R eport 1. O f th e nine LCS sam ples prepared and reported fo r PFOS, m ore than 67% (6 out o f 9) o f the LCS sam ples were within 20% of their respective nominal value. The average recovery for PFOS will be flagged as outside m ethod acceptance criteria. All LCSs were used in the determ ination o f the analytical method uncertainty. Project Lead/PAI Approval: i , Study Director (if GLP): ^ Sponsor Approval (for GLP protocol deviations): NA Date: / i Sbl/^s # -Daf: 2 Date: NA Technical Reviewer (optional): NA Date: NA Laboratory Departm ent Manager Approval: Date: J r: /i~ IV. Authorization to Resume Work Where halting of work occurred, resumption of work must first be approved by Laboratory Management Laboratory Department Manager Approval: NA Date: NA Deviation No. (assigned by Study Director or Team Leader at the end o f study or project) ETS-4-008.7 Page 1 of 1 Documentation of Deviations and Control of Nonconforming Testing Page 74 of 74