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GLP10-01-02: Interim Report 26 - Analysis of PFBS, PFHS, and PFOS in Surface Water Samples Collected at Bert Jeffries Landfill in Decatur, AL in February 2012 Study Title Analysis of Perfluorooctane Sulfonate (PFOS), Perfluorohexane Sulfonate (PFHS) and Perfluorobutane Sulfonate (P FBS ) in G roundwater, Soil and Sedim ent fo r the 3M D ecatur Phase 3 Site-Related Monitoring Program Data Requirement EPA TSC A Good Laboratory Practice Standards 40 CFR Part 792 Study Director Jaisim ha Kesari P.E., DEE W eston Solutions, Inc. 1400 W eston W ay W est Chester, PA 19380 Phone: 610-701-3761 A u th o r Susan W olf 3M Environm ental Laboratory Interim Report Com pletion Date Date of signing Perform ing Laboratory 3M Environmental Health and Safety Operations Environmental Laboratory 3M Center, Bldg 260-05-N-17 St. Paul, MN 55144 Project Identification G L P 1 0 -01-02-26 Total Number o f Pages 77 mm," K The testing reported herein meet the requirements o f ANSI/ISO/IEC 17025:2005 " General Requirements fo r the Competence o f Testing and Calibration Laboratories" , in accordance w ith the A2LA Testing Certificate # (a c cr ed ited ! 2052.01. Testing that complies w ith this International Standard also meets principles o f ISO 9001:2000. Testing Cert #2052.01 This page has been reserved for specific country requirements. GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 GLP C o m p l ia n c e S t a t e m e n t Report Title: Interim Report 26 - A nalysis o f PFBS, PFHS, and PFO S in Surface W a te r Sam ples Collected at Bert Jeffries Landfill in Decatur, A L in February 2012 Study: Analysis of Perfluorooctane Sulfonate (PFOS), Perfluorohexane Sulfonate (PFHS) and Perfluorobutane Sulfonate (PFBS) in Groundwater, Soil and Sedim ent fo r the 3M D ecatur Phase 3 Site-Related Monitoring Program. This analytical phase w as conducted in com pliance with T oxic Substances Control A ct (TSCA) Good Laboratory Practice (G LP) Standards, 40 C FR 792, with the exceptions listed below: These are environm ental sam ples w here 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. Jaisimha Kesari, P.E., DEE, Study Director Date Page 3 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Q u a lity A s s u r a n c e S tatem ent Report Title: Interim Report 26 - A nalysis o f PFBS, PFHS, and PFOS in Surface W a te r Sam ples Collected at Bert Jeffries Landfill in Decatur, A L in February 2012 Study: Analysis o f Perfluorooctane Sulfonate (PFOS), Perfluorohexane Sulfonate (PFHS) and Perfluorobutane Sulfonate (PFBS) in Groundwater, Soil and S edim ent fo r the 3M D ecatur Phase 3 Site-Related Monitoring Program. This analytical phase w as audited by the 3M Environmental Laboratory Quality Assurance Unit (QAU), as indicated in the following table. The findings w ere reported to the principal investigator (P.I.), laboratory m anagem ent and study director. Inspection Dates 4/2/12-4/3/12 Phase Data and Report QAU Representative Date Re sorted to Testing Facility Management Study Director 4/9/12 4/6/12 7- H ' /X Date Page 4 of 77 Ta b le o f C ontents G LP C om pliance S tatem ent..................................................................................................................................3 Q uality A ssurance S tatem ent............................................................................................................................... 4 T able o f C o n te n ts.................................................................................................................................................... 5 List o f T a b le s ............................................................................................................................................................ 6 1 S tudy Info rm a tio n .............................................................................................................................................7 2 S u m m a ry ........................................................................................................................................................... 8 3 Introduction........................................................................................................................................................9 4 T est & Control S u b s ta n c e s ........................................................................................................................... 9 5 R eference S u b s ta n c e s .................................................................................................................................10 6 T est S y s te m ....................................................................................................................................................11 7 M ethod S u m m a ry ..........................................................................................................................................12 7.1 M e th o d s .........................................................................................................................................12 7.2 S am ple C ollection........................................................................................................................ 12 7.3 S am ple Preparation.....................................................................................................................12 7.4 A n a ly s is ......................................................................................................................................... 12 8 A nalytical R e su lts...........................................................................................................................................13 8.1 C a libra tio n ..................................................................................................................................... 13 8.2 System S u ita b ility........................................................................................................................ 14 8.3 Lim it o f Quantitation (L O Q )........................................................................................................14 8.4 Continuing C a libra tio n ................................................................................................................ 14 8.5 B lanks............................................................................................................................................. 14 8.6 Lab Control Spikes (L C S s )....................................................................................................... 14 8.7 Analytical Method U n ce rta in ty.................................................................................................. 16 8.9 Field Matrix Spikes (F M S )............................................................................................................16 Page 5 of 77 9 Data Summary and Discussion 16 10 C on clusion...................................................................................................................................................... 20 11 D ata/Sam ple R e te n tio n .........'..................................................................................................................... 20 12 A tta c h m e n ts ................................................................................................................................................... 20 13 S ig n a tu re s ...................................................................................................................................................... 21 L is t o f Ta b l e s Table 1. S um m arized PFBS, PFHS, and P FO S Results (Jeffries Landfill Site, Feb 2012)......8 T able 2. S am ple Description Key C ode.............................................................................................. 11 Table 3. Instrum ent P aram eters................................................ 12 Table 4. Liquid C hrom atography C onditions......................................................................................13 T able 5. M ass Transitions...................................................................................................................... 13 T able 6. Lim it o f Q uantitation (LO Q ).................................................................................................... 14 Table 7. Laboratory Control Spike R ecovery..................................................................................... 15 Table 8. Analytical U ncertainty.............................................................................................................. 16 T able 9. Field M atrix Spike Levels........................................................................................................16 Table 10. JP A L S W ET01 120228.......... ............................................................................................17 Table 11. JP A L S W E T 0 2 120228........................................................................................................ 17 Table 12. JP A L S W ET03 120228........................................................................................................ 18 Table 13. JP A L S W ET04 120228........................................................................................................ 18 Table 14. Trip Blank 1 2 0 2 2 8 .................................................................................................................19 Page 6 of 77 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 W eston Solutions, Inc. W est 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 W illiam K. Reagen, Ph.D., 3M Laboratory M anager Cleston Lange, Ph.D., Principal Analytical Investigator, (clanqe@ mmm.com1: phone (651)-733-9860 Susan Wolf, 3M Analyst Chelsie Grochow, A nalyst Kevin Eich, Analyst Kelly Ukes, A nalyst Jonathan Steege, Analyst Study Dates Study Initiation: March 8, 2010 Interim 26 Experim ental Termination: March 14, 2012 Interim Report Completion: Date o f Interim Report Signing Location o f Archives All original raw data and the analytical report have been archived at the 3M Environm ental Laboratory according to 40 C FR Part 792. The test substance and analytical reference standard reserve sam ples are archived at the 3M Environm ental Laboratory according to 40 CFR Part 792 Page 7 of 77 2 Summary The 3M Environmental Laboratory received surface water samples from four different locations located at Jeffries Landfill in Decatur, AL. A total o f nineteen sam ple bottles w ere received at the 3M Environmental Laboratory for perfluorooctane sulfonate (PFOS), perfluorohexane sulfonate (PFHS) and perfluorobutane sulfonate (PFBS), and included duplicate surface w ater samples and two field matrix spike (FMS) samples from each sampling location. A set o f trip blank samples; single trip blank containing M illi-Q TM water and two trip blank spikes, were included with the sample bottles. All samples were logged into the laboratory information m anagem ent system (LIMS) under project GLP10-01-0226. The surface w ater sam ples and trip blanks w ere received from W eston personnel on March 2, 2012. All o f the samples were prepared and analyzed for PFBS, PFHS, PFOS, and the surrogate recovery standard 13C4-PFO S, following 3M Environm ental Laboratory Method ETS-8-044.1. The average m easured PFBS, PFHS, and PFO S concentrations are sum m arized in Table 1. T he trip blank sam ple was below the lower limit o f quantitation (LLO Q ) fo r all analytes, indicating adequate control of sample contamination during shipping and sample collections. The analytical method uncertainties associated with the reported results are: PFBS + 22%, PFHS + 19% and PFOS + 15%. Table 1. Summarized PFBS, PFHS, and PFOS Results (Jeffries Landfill Site, Feb 2012). 3M LIMS ID GLP10-01-02-026-005 GLP10-01-02-026-006 GLP10-01-02-026-009 GLP10-01-02-026-010 GLP10-01-02-026-013 GLP10-01-02-026-014 GLP10-01-02-026-017 GLP10-01-02-026-018 GLP10-01 -02-026-025 Sample Description JPAL-SW-ET01-0-120228 JPAL-SW-ET01-DB-120228 Average %RPD Sample/Sample Dup JPAL-SW-ET02-0-120228 JPAL-SW-ET02-DB-120228 Average %RPD Sample/Sample Dup JPAL-SW-ET03-0-120228 JPAL-SW-ET03-DB-120228 Average %RPD Sample/Sample Dup JPAL-SW-ET04-0-120228 JPAL-SW-ET04-DB-120228 Average %RPD Sample/Sample Dup JPAL-SW-TRIP1-0-120228 PFBS Concentration (ng/mL) 0.0345 0.0362 0.0354 4.8 <0.0250 <0.0250 <0.0250 NA <0.0250 <0.0250 <0.0250 NA <0.0250 <0.0250 <0.0250 NA <0.0250 PFHS Concentration (ng/mL) <0.0250 <0.0250 <0.0250 NA <0.0250 <0.0250 <0.0250 NA <0.0250 0.0252 0.0252 NATM <0.0250 <0.0250 <0.0250 NA <0.0250 PFOS Concentration (ng/mL) 0.0302 0.0302 0.0302 0.0 0.0661 0.0593 0.0627 11 0.303 0.303 0.303 0.0 0.166 0.159 0.163 4.3 <0.0232 NA = Not Applicable The analytical method uncertainties associated with the reported results are: PFBS 22%, PFHS 19 %, and PFOS 15%. (1) A sample/sample duplicate RPD could not be determined since the concentration for the primary sample was BLOQ for PFHS. Page 8 of 77 3 Introduction This analytical study was conducted as part of the Phase 3 Environmental Monitoring and Assessm ent Program fo r the 3M facility located in Decatur, Alabam a. T he objective o f the overall program is to gain information regarding concentrations of perfluorooctane sulfonate (PFOS), perfluorohexane sulfonate (PFHS) and perfluorobutane sulfonate (PFBS), in various environm ental media such as groundwater, soils and sedim ents that are associated with and near the Decatur facility. This analytical study was conducted to analyze surface w ater sam ples collected from fo u r locations at o r near Jeffries Landfill in Decatur, A L fo r PFBS, PFHS, and PFO S in an effort to characterize surface w ater conditions. The 3M Environmental Laboratory prepared sample containers (250 m L high-density polyethylene bottles) which were shipped to Decatur, AL W eston personnel prior to field sampling. Sample containers for each sampling location included a field sample, field sam ple duplicate, and tw o field m atrix spike sam ples. Each em pty container w as m arked with a "fill to here" line to produce a final sample volum e o f 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 o f 180 2-PFBS, 13C3-PFHS, and 13C8-P FO S (internal standard) a t a nominal concentration o f 1 ng/m L. All sam ple bottles also included the addition o f 13C4-PFO S (surrogate recovery standard) at a nominal concentration o f 0.1 ng/m L. See section 8.8 o f the report fo r field matrix spike levels. Sam ples w ere prepared and analyzed according to the procedure defined in 3M Environm ental Laboratory method ETS-8-044.1 "Method o f Analysis for the Determination o f Perfluorinated C om pounds In W a te r by LC/MS/M S; Direct Injection A nalysis" . T he use o f internal standards w as used to aid in the data quality objectives. Table 1 sum m arizes the average PFBS, PFHS, and PFO S concentrations fo r the duplicate surface w ater sam ples collected and the trip blank sample. Tables 10-14 sum m arize the individual sam ple results and the associated field matrix spike recoveries. All results for the quality control samples prepared and analyzed with the sam ples are reported and discussed elsewhere in this report 4 Test & Control Substances There w as not a test substance o r control substances in the classic sense o f a GLP study. This study w as purely analytical in nature. Page 9 of 77 5 Reference Substances Reference Substance Chemical Name Chemical Formula Identifier Use Source Expiration Date Storage Conditions Chemical Lot Number TCR Number Physical Description Purity PFBS (predominantly linear) Potassium Perfluorobutane sulfonate c 4f 9s o 3x + NA Target Analyte Reference Standard 3M 01/10/2017 Frozen 41-2600-8442-5 TCR-121 White Powder 96.7% '"Oj-PFBS [1802]-Ammonium Perfiuorobutanesulfonate C4F9S [1802]0-N H 4+ NA Internal Standard RTI International 03/09/2015 Frozen 11546-107-2 TCR-1044, TCR-1040 Liquid >99% Reference Substance Chemical Name Chemical Formula Identifier Use Source Expiration Date Storage Conditions Chemical Lot Number TCR Number Physical Description Purity PFHS (Linear) Sodium Perfluorohexane sulfonate C 6F1 3 S O 3 Na L-PFHXS Target Analyte Reference Standard Wellington 03/25/2018 Frozen LPFHxSAM08 TCR08-0018 Crystalline 100% 13C3-PFHS Sodium Perfluorohexanesulfonate 13C312C3F13S 0 3 -Na+ MPFC-C-0511 Internal Standard Wellington 05/25/2014 Frozen MPFC-C-0511 TCR11-0016 Liquid 5 pg/mL(1> 13C3-PFHS Sodium Perfluorohexanesulfonate 13C312C 3F13S0 3 -Na+ MPFC-C-0112 Internal Standard Wellington 01/24/2015 Frozen MPFC-C-0112 TCR12-0004 Liquid 5 pg/mL<1) (1) Custom mixture of seven mass-labeled (13C) perfluoroalkylcarboxylic acids, two mass labeled (13C) perfluoroalkylsulfonates and one mass-labeled (13C) perfluoro-1-octanesulfonamide. Page 10 of 77 Reference Substance Chemical Name Chemical Formula Identifier Use Source Expiration Date Storage Conditions Chemical Lot Number TCR Number Physical Description Purity PFOS (Linear + Branched) Potassium Perfluorooctane sulfonate C8F17S 0 3'K + Br-PFOSK Target Analyte Reference Standard Wellington 03/17/2014; 12/01/2014 Frozen brPFOSK0708; brPFOSK1111 TCR11-0010; TCR11-0041 Liquid 99.9% 13c4-pfos Sodium Perfluorooctane sulfonate 13C412C4F17S 0 3'N a+ MPFOS Surrogate Wellington 09/08/2013 Frozen MPFOS0910 TCR10-0044-7/9 Liquid >98% 13C,,-PFOS Sodium Perfluorooctane sulfonate 13C8F 17S 0 3'N a+ MPFC-C-0511 Internal Standard Wellington 05/25/2014 Frozen MPFC-C-0511 TCR11-0016 Liquid 5 p g /m L (1> 13Ca-PFOS Sodium Perfluorooctane sulfonate 13C8F 17S 0 3'N a+ MPFC-C-0112 Internal Standard Wellington 01/24/2015 Frozen MPFC-C-0112 TCR12-0004 Liquid 5 pg/mL(1> (1) Custom mixture of seven mass-labeled (13C) perfluoroalkylcarboxylic adds, two mass labeled (13C) perfluoroalkylsulfonates and one mass-labeled (13C) perfluoro-1-octanesulfonamlde. 6 Test System The test system s fo r this study are surface w ater sam ples collected from wells located in Decatur, A L by W eston Solutions, Inc. personnel. S am ples fo r this study are "real w orld" samples, not dosed with a specific lot of test substance. Table 2. Sample Description Key Code. String Number Example 1 2 3 4 5 String Descriptor JPAL-SW-ET01-0-120228 Sample Location Sample Type Well ID Sampling Date Sample Type Example JPAL= Jeffries Property, Alabama SW = Surface Water Example: ET01 E T = Eastern tributary to Mallard Creek 120228- Febmary 28, 2012 0 - primary sample DB = duplicate sample LS = low spike HS = high spike Page 11 of 77 7 Method Summary 7.1 M e th o d Analysis fo r all analytes w as com pleted following 3M Environm ental Laboratory method ETS-8-044.1 "Method o f Analysis fo r the Determ ination o f Perfluorinated C om pounds In W a te r by High Perform ance Liquid Chromatography/Mass Spectrom etry Direct Injection Analysis". 7.2 Sam ple C o lle ctio n S am ples w ere collected in 250 m L N a lg e n e TM (high-density polyethylene) bottles prepared at the 3M Environmental Laboratory. Sample bottles associated with G LP10-01-02-26 were returned to the laboratory at ambient conditions on March 2, 2012. Samples w ere stored refrigerated at the laboratory after receipt. A set of laboratory prepared Trip Blank and Trip Blank field matrix spikes were sent with the sample collection bottles. 7.3 Sam ple P reparation S am ples w ere prepared by rem oving an aliquot o f the well mixed sam ple and placing it in an autovial for analysis. During the preparation o f the laboratory control samples, an aliquot o f a separate internal standard spiking solution w as added to the laboratory control samples (nominal concentration o f 1 ng/mL). The samples bottles were spiked with an internal standard m ix at a nominal concentration o f 1 ng/m L prior to being sent to the field fo r sam ple collection 7.4 A n alysis All study sam ples and quality control samples w ere analyzed fo r PFBS, PFHS, and PFOS using high perform ance liquid chrom atography/ tandem m ass spectrom etry (HPLC/M S/M S). Detailed instrum ent param eters, the liquid chrom atography gradient program , and the specific m ass transitions analyzed are described in the raw data hard copies placed in the final data packet, and are briefly described below. Table 3. Instrum ent Parameters. Instrument Name Analytical Method Followed Analysis Date Liquid Chromatograph Guard column Analytical column Injection Volume Mass Spectrometer Ion Source Electrode Polarity Software ETS Ginger ETS-8-044.1 3/7/12-P F B S and PFHS Agilent 1100 Betasil C18 (4.6 mm X 100 mm), 5g Betasil C18 (4.6 mm X 100 mm), 5ji 25 uL Applied Biosystems API 5000 Turbo Spray Turbo ion electrode Negative Analyst 1.4.2 ETS McCoy ETS-8-044.1 3/13/12-P FO S Agilent 1290 Betasil C18 (4.6 mm X 100 mm), 5u Betasil C18 (4.6 mm X 100 mm), 5p 50 pL Applied Biosystems API 5500 Turbo Spray Turbo ion electrode Negative Analyst 1.5.2 Page 12 of 77 Table 4. Liquid Chromatography Conditions. Step Number 0 1 2 3 4 5 Step Number 0 1 2 3 4 5 Total Time (min) 0.0 ` 2.0 14.5 15.5 16.5 20.0 Total Time (min) 0.0 2.0 14.5 15.5 16.5 20.0 Flow Rate (juL/min) Percent A (2 mM ammonium acetate) ETS-8-044.1 Analyzed 3/7/12 750 97.0 750 97.0 750 5.0 750 5.0 750 97.0 750 97.0 Flow Rate (fdJmin) Percent A (5 mM ammonium acetate: 0.01% acetic add) ETS-8-044.1 Analyzed 3/13/12 750 97.0 750 97.0 750 5.0 750 5.0 750 97.0 750 97.0 PercentB (methanol) 3.0 3.0 95.0 95.0 3.0 3.0 Percent B (acetonitrile) 3.0 3.0 95.0 95.0 3.0 3.0 Table 5. Mass Transitions. Analyte PFBS PFHS PFOS FCJ-PFOS Mass Transition Q1/Q3 299/80 299/99 399/80 399/99 499/80 499/99 499/130 503/80 Reference Material Structure Linear Linear Linear + Branched Linear Internal Standard f sOrf-PFBS f 3CJ-PFHS f 3Ce]-PFOS fC J-P F O S Mass Transition Q1/Q3 303/84 402/80 507/80 507/80 Dwell time was 50 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 C a lib ra tio n Samples were analyzed using a stable isotope internal standard calibration curve. Calibration standards were prepared by spiking known am ounts of the stock solution containing the target analytes into a laboratory-prepared synthetic groundwater containing calcium and magnesium. A separate internal standard spiking solution w a s prepared and an aliquot w as added at the sam e level to all calibration standards and laboratory control samples at a nominal concentration o f 1 ng/mL. A calibration curve ranging from approxim ately 0.025 ng/m L to 100 ng/m L (0.025 ng/m L to 10 ng/m L fo r 13C4-PFO S surrogate) was prepared. The reference standard used for the calibration standards for PFOS contained both linear and branched isomers. Page 13 of 77 A quadratic, 1/x weighted, calibration curve of the standard peak area/peak area ratios w as used to fit the data for each analyte. The data w ere not forced through zero during the fitting process. Calculating the standard concentrations using the peak area/peak area ratios and the resultant calibration curve confirm ed accuracy o f each curve point. Each curve point was quantitated using the overall calibration curve and reviewed for accuracy. Method calibration accuracy requirem ents o f 10025% (10030% fo r the lowest curve point) w ere m et fo r all analytes. The correlation coefficient (r) w as greater than 0.995 fo r PFBS, PFHS, PFOS, and 13C4-PFOS. 8.2 S ystem S u ita b ility A calibration standard w as analyzed four times at the beginning of each analytical sequence to dem onstrate overall system suitability. The acceptance criteria o f less than or equal to 5% relative standard deviation (RSD) for peak area and retention tim e criteria o f less than or equal to 2% RSD was m et fo r PFBS, PFHS, PFOS, and 13C4-PFOS. 8.3 L im it o f Q uantitatio n (LOQ) The LOQ fo r this analysis is the lowest non-zero calibration standard in the curve that m eets linearity and accuracy requirements and for which the area counts or area ratio are at least twice those o f the appropriate blanks. T he LO Q fo r all analytes can be found in Table 6. Table 6. Lim it o f Q uantitation (LOQ). PFBS LOQ, ng/mL 0.0250 PFHS LOQ, ng/mL 0.0250 PFOS LOQ, ng/mL 0.0232 8.4 C o ntinu ing C alibration During the course o f each analytical sequence, continuing calibration verification samples (CCVs) were analyzed to confirm that the instrum ent response and the initial calibration curve were still in control. All CCVs m et method criteria o f 100% 25% fo r PFBS, PFHS, PFOS, and 13C4-PFOS. 8.5 B lanks Tw o types o f blanks were prepared and analyzed w ith the samples: procedural blanks and trip blanks. Procedural blank results were reviewed and used to evaluate method performance to determine the LOQ. Trip blanks reflect the shipping and sample collection conditons the sample bottles and sam ples experience. 8.6 Lab C ontrol S pikes (LCSs) Low, mid, and high lab control spikes w ere prepared fo r the target analytes and analyzed in triplicate, while only low and high lab control spikes w ere prepared fo r the 13C4-P FO S surrogate. LCSs w ere prepared by spiking known am ounts o f the analyte into synthetic groundw ater to produce the desired concentration. The spiked w ater sam ples w ere then prepared and analyzed in the sam e m anner as the samples. The method acceptance criteria, average of LCS at each level should be within 100% 20% with an RSD <20% , w as m et fo r all analytes. T he following calculations w ere used to generate data in T able 7 fo r laboratory control spikes. ,LC_ _S P_ ercent. ,,Recovery = -C--a--l-c-u--l-a--t-e-d---C--o--n--c--e-n--t-r-a--t-i-o-n-- *, 1. 0. .0.%. Spike Concentration LCS% RSD = standard deviation LCS replicates , 1Q0% average LCS recovery Page 14 of 77 Table 7. Laboratory Control Spike Recovery. ETS-8-044.1 Analyzed 3/7/12 Lab ID Spiked Concentration (ng/mL) PFBS Calculated Concentration (ng/mL) %Recovery Spiked Concentration (ng/mL) LCS-120306-1 LCS-120306-2 LCS-120306-3 Average %RSD 0.198 0.198 0.198 0.209 0.209 0.207 105% 0.95% 105 106 104 0.198 0.198 0.198 LCS-120306-4 1.98 2.17 LCS-120306-5 1.98 2.19 LCS-120306-6 1.98 2.09 Average %RSD 109% 2.4% 110 111 106 1.98 1.98 1.98 LCS-120306-7 19.9 18.9 LCS-120306-8 19.9 18.1 LCS-120306-9 19.9 19.8 Average %RSD 95.3% 4.5% 95.1 91.1 99.6 19.8 19.8 19.8 PFHS Calculated Concentration (ng/mL) 0.191 0.182 0.185 93.8% 2.4% 2.08 1.92 1.94 100% 4.3% 18.5 18.1 19.2 93.9% 3.0% YoRecovery 96.3 91.8 93.3 105 97.1 98.0 93.5 91.3 96.9 ETS-8-044.1 Analyzed 3/13/12 Lab ID PFOS (Linear + Branched) 13C4-PFOS surrogate Spiked Calculated Spiked Concentration Concentration Concentration (ng/mL) (ng/mL) YoRecovery (ng/mL) Calculated Concentration (ng/mL) YoRecovery LCS-120306-1 LCS-120306-2 LCS-120306-3 Average %RSD 0.184 0.184 0.184 0.178 0.170 0.170 94.1% 2.7% 97.0 92.6 92.6 0.189 0.189 0.189 0.221 0.215 0.225 117% 2.2% 117 114 119 LCS-120306-4 1.84 1.76 LCS-120306-5 1.84 1.72 LCS-120306-6 1.84 1.71 Average %RSD 94.0% 1.5% 95.6 93.6 92.9 1.89 2.21 1.89 2.14 1.89 2.14 114% 2.0% 117 113 113 LCS-120306-7 18.4 17.1 92.8 NA NA NA LCS-120306-8 18.4 16.6 90.4 NA NA NA LCS-120306-9 18.4 17.4 94.3 NA NA NA Average %RSD 92.5% 2.1% NA NA = Not Applicable Page 15 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 8.7 A n a lytica l M ethod U n certainty 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 fo r the set o f accuracy results (in %) obtained fo r the Q C samples. The expanded uncertainty is calculated by multiplying the standard deviation by a factor o f 2, which corresponds to a confidence level o f 95%. Table 8. A nalytical Uncertainty. Analyte PFBS PFHS PFOS Standard Deviation 10.8 9.42 7.60 Method Uncertainty 22% 19% ' 15% 8.8 Field M atrix S pikes (FMS) Low, mid, and high field matrix spikes (FMS) w ere collected at each sampling point to verify that the analytical method is applicable to the collected matrix. Field m atrix spikes w ere generated by adding a measured volum e o f field sam ple to a container spiked by the laboratory with PFBS (linear), PFHS (linear), and PFOS (linear+branched) prior to shipping sam ple containers fo r sample collection. Field matrix spike recoveries within method acceptance criteria o f 10030% confim i that "unknown" com ponents in the sam ple m atrix do not significantly interfere with th e extraction and analysis o f the analytes o f interest. Field m atrix spike concentrations m ust be 50% o f the sam ple concentration to be considered an appropriate field spike. Field m atrix spikes are presented in section 9 o f this report. Table 9. Field M atrix Spike Levels. Sampling Location Spike Level All locations and Trip Blank Low High PFBS, ng/mL 1.00 10.0 PFHS, ng/mL 0.998 9.98 PFOS, ng/mL 0.928 9.28 FMS Recovery ( Sample Concentration of FMS - Average Concentration: Field Sample & Field Sample Pup.) * 10go/ Spike Concentraton 9 Data Summary and Discussion The tables below sum m arize the sam ple results and field 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 o f rounding, values m ay vary slightly from those listed in the raw data. Field matrix spike recoveries meeting the method acceptance criteria of 30%, dem onstrate that the method was appropriate for the given matrix and their respective quantitative ranges. Page 16 of 77 Table 10. JPAL SW ET01 120228 3M L IM S ID Description GLP10-01-02-26-005 JPAL-SW-ET01-0-120228 GLP10-01-02-26-006 JPAL-SW-ET01-DB-120228 GLP10-01-02-26-007 JPAL-SW-ET01-LS-120228 GLP10-01-02-26-008 JPAL-SW-ET01-HS-120228 Average Concentration (ng/mL) t %RPD NA = Not Applicable PFBS PFHS PFOS Concentration (ng/mL) %Recovery 0.0345 0.0362 1.08 10.4 NA NA 104 104 0.0354 ng/mL 4.8% Concentration (ng/mL) %Recovery <0.0250 <0.0250 0.946 8.62 NA NA 94.8 86.4 <0.0250 ng/mL Concentration (ng/mL) %Recovery 0.0302 NA 0.0302 0.863 8.38 NA 89.7 90.0 0.0302 ng/mL 0.0% 13C4 PFOS %Recovery 109 102 110 110 108% 3.3% Table 11. JPAL SW ET02 120228 3 M L IM S ID Description GLP10-01-02-26-009 JPAL-SW-ET02-0-120228 GLP10-01 -02-26-010 JPAL-SW-ET02-DB-120228 GLP10-01-02-26-011 JPAL-SW-ET02-LS-120228 GLP10-01-02-26-012 JPAL-SW-ET02-HS-120228 Average Concentration (ng/mL) %RPD NA = Not Applicable PFBS PFHS PFOS Concentration (ng/mL) %Recovery <0.0250 NA <0.0250 1.08 9.63 NA 108 96.3 <0.0250 ng/mL Concentration (ng/mL) %Recovery <0.0250 <0.0250 NA NA 1.030 8.640 103 86.6 <0.0250 ng/mL Concentration (ng/mL) %Recovery 0.0661 0.0593 0.924 8.45 NA NA 92.8 90.4 0.0627 ng/mL 11% 13C4 PFOS %Recovery 107 106 109 107 107% 1.2% Page 17 of 77 Table 12. JPAL SW ET03 120228 PFBS PFHS PFOS 3M U M S ID Description Concentration (ng/mL) %Recovery GLP10-01 -02-26-013 JPAL-SW-ET03-0-120228 . GLP10-01-02-26-014 JPAL-SW-ET03-DB-120228 GLP10-01 -02-26-015 JPAL-SW-ET03-LS-120228 GLP10-01 -02-26-016 JPAL-SW-ET03-HS-120228 Average Concentration (ng/mL) %RPD <0.0250 NA <0.0250 NA 1.05 105 9.14 91.4 <0.0250 ng/mL Concentration (ng/mL) YoRecovery <0.0250 NA 0.0252 NA 0.992 96.9 8.820 88.1 0.0252 ng/mL<1> Concentration (ng/mL) YoRecovery 0.303 0.303 1.13 8.72 NA NA 89.1 90.7 0.303 ng/mL i O.OYo 13c4 p f o s YoRecovery 109 109 103 106 107%2.7% NA = Not Applicable (1) A sample/sample duplicate RPD could not be determined since the concentration o f the primary sample was BLOQ for PFHS. Table 13. JPAL SW ET04 120228 3 M L IM S ID Description GLP10-01-02-26-017 JPAL-SW-ET04-0-120228 GLP10-01-02-26-018 JPAL-SW-ET04-DB-120228 GLP10-01-02-26-019 JPAL-SW-ET04-LS-120228 GLP10-01 -02-26-020 JPAL-SW-ET04-HS-120228 Average Concentration (ng/mL) YoRPD NA = Not Applicable PFBS PFHS PFOS Concentration (ng/mL) YoRecovery <0.0250 NA <0.0250 NA 1.14 114 10.6 106 <0.0250 ng/mL Concentration (ng/mL) YoRecovery <0.0250 <0.0250 0.931 8.45 NA NA 93.3 84.7 <0.0250 ng/mL Concentration (ng/mL) YoRecovery 0.166 0.159 1.03 8.47 NA NA 93.5 89.5 0.163 ng/mL 4.3%, 13C4 PFOS YoRecovery 113 109 110 105 109% 3.2% Page 18 of 77 Table 14. Trip Blank 120228 3 M L IM S ID Description GLP10-01-02-26-025 JPAL-SW-TRIP1 -0-120228 GLP10-01-02-26-026 JPAL-SW-TRIP1 -LS-120228 GLP10-01-02-26-027 JPAL-SW-TRIP1 -HS-120228 PFBS PFHS PFOS Concentration (ng/mL) <0.0250 1.06 9.32 %Recovery NA 106 93.2 Concentration (ng/mL) <0.0250 0.947 9.21 %Recovery NA 94.9 92.3 Concentration (ng/mL) <0.0232 0.809 8.07 %Recovery NA 87.2 87.0 13C4 PFOS %Recovery 108 109 107 NA = Not Applicable Page 19 of 77 10 Conclusion Laboratory control spikes and field matrix spikes were used to determine the analytical method accuracy and precision for PFBS, PFHS, and PFOS. Analysis was successfully completed following 3M Environmental Laboratory method ETS-8-044.1 described herein. 11 Data/Sample Retention All remaining sam ples and associated project data (hardcopy and electronic) will be archived according to 3M Environmental Laboratory standard operating procedures. 12 Attachments Attachm ent A: Protocol Am endm ent 26 (General Project Outline) A ttachm ent B: Representative Chrom atogram s and Calibration Curves Attachm ent C: Analytical Method-ETS-8-044.1 Page 20 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 13 Signatures Cleston Lange, Ph.D., 3M Principal Analytical Investigator U / Z a j Z. Date ---------- ----------------------------------------------------- ^ W illiam K. Reagen, Ph.D., 3M Environmental Laboratory Departm ent Manager Date Page 21 of 77 Attachment A : Protocol A mendment GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Page 22 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Analytical Protocol: GLP10-01-02 Amendment 26 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. 26 Amendment Date: February 21,2012 Performing Laboratory 3M Environmental, Health, and Safety Operations 3M Environmental Laboratory Building 260-5N-17 Maplewood, MN 55144-1000 Laboratory Project identification GLP10-01-02-26 Sampling Event Bert Jeffries Landfill -Surface Water Sampling Page 1 of 6 Page 23 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Analytical Protocol: GLP10-01-02 Amendment 26 This amendment modifies die following portion o f 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. AMEND TO READ: No changes to the wording of the protocol are required. This amendment only addresses and documents the addition o f the General Project Outline (GPO) for the collection and analysis o f groundwater samples from Decatur, AL, and conducted as part of the 3M Decatur Phase 3 Program for PFOS, PFHS and PFBS (GLP10-01-02). Sampling activities at and near the Bert Jeffries landfill are scheduled for the week of February 27, 2012 and will consist of 6 paired surface water and sediment samples. This protocol. amendment addresses the sampling and analysis of the surface w ater samples. Surface water samples collected under this sampling event will be entered into the 3M Environmental Laboratory LI MS as project GLP10-01-02-26 and reported as interim report GLP10-01-02-26 (reflecting study GLP10-01-02 and amendment -26). Reason: The reason for this amendment is to document the General Project Outline (GPO) which describes the anticipated surface water sample collection event to be conducted at and near the Bert Jeffries landfill in Lawrence County, AL. The GPO is three pages in length and included as attached to this amendment form. Page 2 of 6 Page 24 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Analytical Protocol: GLP10-01-02 Amendment 26 Amendment Approval Page 3 of 6 Page 25 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Analytical Protocol: GLP10-01-02 Amendment 26 Environmental Health & Safety Operations, Environmental Laboratory General Project Outline To: From: cc: Date: S u b je c t: Gary Hohenstein, 3M EHS&Opns Susan Wolf, 3M EHS&Opns; Environmental Lab William Reagen, 3M EHS&Opns; Environmental Lab Jai Kesari, Weston Solutions Charles Young, Weston Solutions February 21,2012 Analysis of Perfluorooctane Sulfonate (PFOS), Perfluorohexane Sulfonate (PFHS) and Perfluorobutane sulfonate (PFBS) in Groundwater, Soil and Sediment for the 3M Decatur Phase 3 Site-Related Monitoring Program; GLP Interim Report 26 - Bert Jeffries Landfill; Surface W ater sampling the week of February 27, 2012. 1 General Project Information Contacts Lab Request Number Six Digit Department Number Project $hedule/Test Dates 3M Sponsor Representative Gary Hohenstein 3M EHS Operations 3M Building 224-5W-03 Saint Paul, MN 55144-1000 Phone: (651) 737-3570 aahohenstein@mmm:com 3M Environmental Laboratory Management William K. Reagen 3M EHS Opns, Environmental Laboratory 260-5N-17 651 733-9739 wkreaaen@mmm.com Principal Analytical Investigator Cleston Lange 3M EHS Opns, Environmental Laboratory 260-5N-17 651 733-9860 cclanae@mmm.com Sampling Coordinator Timothy Frinak Weston Solutions Timothv.frinaK@westonsolulions.cQm Phone: (3341-332-9123 . GLP10-01-02-26 Dept #530711, Project #0022674449 Tentatively scheduled for the week of February 2 7 ,2Q12. All verbal and written correspondence will be directed to Gary Hohenstein. Page 4 of 6 Page 26 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Analytical Protocol: GLP10-01-02 Amendment 26 2 Background Information and Project Objective(s) The 3M EHS Operations Laboratory (3M Environmental Lab) will receive and analyze surface water samples collected from six (6) locations at and near the Bert Jeffries landfill. Surface water samples will be collected by Weston. The 3M Environmental Laboratory will prepare the sample bottles; Samples will be analyzed for Perfiuorooctane Sulfonate (PFOS), Perfluorohexane Sulfonate (PFHS) and Perfluorobutane Sulfonate (PFBS). Analyses will be conducted under the GLP requirements o f EPA TSCA Good Laboratory Practice Standards 40 CFR 792. The final report will be submitted to Gary Hohenstein and Jai Kesari upon completion under interim report GLP10-01-02-26. 3 Project Schedule ________________________ Sample collection bottles will be prepared by the 3M Environmental Laboratory. Sample bottles will be shipped in coolers overnight to 3M Decatur for arrival by Friday, February 24,2012. Sample bottles should be stored refrigerated on-site until sample collection. Martin Smith \ Weston Trailer 3M Decatur Plant 1400 State Docks Road Decatur, Alabama 35601 4 Test Parameters The targeted limit of quantitation will be 0.025 ng/mL (ppb) for PFBS, PFHS, and PFOS. A total of six sampling locations have been specified. For each sampling location, a total of four sample bottles will be collected (sample, sample duplicate, low-level field matrix spike, and high-level field matrix spike). The "fill to here" line on each 250 mL Nalgene bottle will be 200 mL. One set of trip blanks consisting of reagent-grade water, a low-level trip blank spike, and a high-level trip blank spike will be prepared at the 3M Environmental Laboratory and sent to the sampling location with the other bottles. No previous data on fluorochemicals in groundwater, surface water or sediment media exist for the site. The low field matrix spike will be prepared at 1 ng/mL and the high field matrix spike at 10 ng/mL. All sample bottles will include the addition of 19OrPFBS, 13Cr PFHS, and 13Cs-PFOS, (internal standards) at a nominal concentration of 1 ng/mL. All sample bottles will also include the addition of 13C4-PFOS (surrogate recovery standard) at a nominal concentration of 0.1 ng/mL, To aid in sample collection, a non-spiked 1-L sample bottle will be provided for each sampling location. Sample collection will consist of direct immersion. Aliquots will be decanted from the collection bottle to the primary, duplicate and matrix spike bottles. No rinseate blank will be collected. I Page 5 of 6 Page 27 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 t Analytical Protocol: GLP10-01-02 Amendment 26 5 Test Methods Samples will be prepared and analyzed by LC/MS/MS following ETS-8-044.1 "Method of Analysis for the Determination of Perfluorinated Compounds In Water by High Performance Liquid Chromatography/Mass Spectrometry Direct Injection Analysis". Alternately, samples may be analyzed by ETS 8-154.3 "Determination of Perfluorinated Acids, Alcohols, Amides, and Sulfonates In Water By Solid Phase Extraction and High Performance Liquid Chromatography/Mass Spectrometry". Where applicable, samples will be analyzed against an internal standard calibration curve. Each curve point will contain isotopically-labeled PFBS, PFHS, and PFOS at a nominal concentration o f 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. Laboratory control samples prepared with the samples must have an average recovery within 10020% and a RSD <20%. The data quality objective for this study is quantitative results for the target analytes with an analytical accuracy of 100+30%. Field matrix spikes not yielding recoveries within 100+30% will be addressed in the report and the final accuracy statement may be adjusted accordingly. 6 Reporting Requirements_________________________ For each sampling location, the report will contain the results for the sample, sample duplicate, and the two field matrix spikes. Trip blank and trip blank spikes 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. For those sampling locations where the field matrix spike level was not appropriate due to higher than expected analyte concentrations, a target analyte laboratory matrix spike may be prepared and will be included in the final report Page. 8 of 6 Page 28 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 A t t a c h m e n t B : R e p r e s e n ta t iv e S a m p l e C h r o m a t o g r a m s a n d C a lib r a t io n C u r v e (s ) Page 29 of 77 *ETS-McCoy GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Results Name: mcl20313a.rdb Printing Date: Wednesday, April 11, 2012 Page 30 of 77 *ETS-McCoy GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Results Name: mcl20313a.rdb Printing Date: Wednesday, April 11, 2012 Page 31 of 77 *** Ginger AG01330509 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Results Name: gl20307a.rdb Printing Date: Wednesday, April 11, 2012 Page 32 of 77 *** Ginger AG01330509 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Results Name: gl20307a.rdb Data printed by STW Printing Time: 12:24:26 PM Printing Date: Wednesday, April 11, 2012 Page 1 of 1 Page 33 of 77 *ETS-McCoy GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Results Name mcl20313a.rdb Printing Time 11:34:46 AM Printing Date Wednesday, April 11 2012 Page 34 of 77 *ETS-McCoy GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Results Name mcl20313a.rdb Printing Time: 11:34:46 AM Printing Date: Wednesday. April 11 2012 Page 35 of 77 *ETS-McCoy GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Results Name mcl20313a.rdb Printing Date: Wednesday, April 11, 2012 Page 36 of 77 ETS-McCoy Data printed by STW Printing Time: 11:34:46 AM Printing Date: Wednesday, April 11, 2012 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Results Name mcl20313a.rdb Page 4 of 9 Page 37 of 77 ETS-McCoy Data printed by STW Printing Time: 11:34:46 AM Printing Date: Wednesday, April 11, 2012 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Results Name: mcl20313a.rdb Page 5 of 9 Page 38 of 77 ETS-McCoy Data printed by STW Printing Time: 11:34:46 AM Printing Date: Wednesday, April 11, 2012 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Results Name: mcl20313a.rdb Page 39 of 77 ETS-McCoy Data printed by STW Printing Time: 11:34:46 AM Printing Date: Wednesday, April 11, 2012 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Results Name: mcl20313a.rdb Page 7 of 9 Page 40 of 77 ETS-McCoy Data printed by STW Printing Time: 11:34:46 AM Printing Date: Wednesday, April 11, 2012 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Results Name mcl20313a.rdb Page 8 of 9 Page 41 of 77 *ETS-McCoy GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Results Name: mcl20313a.rdb Printing Date Wednesday, April 11 2012 Page 42 of 77 *** Ginger AG01330509 I Sample Name: *g120307a014* Sample 10.'11012-6-6* Fie: *g120307a.wifr Peak Name *13C3PFHS-IS(ISr Masses). '402.0/80.0 amu' Comment *0.25 ng'mL FC std In Synth. Water* Annotation ** Sample Index: 14 Sample Type: Standard Concentration: 0.945 r Calculated Cone. N/A Acq. Date- 3/7/2012 Acq. Tine: 9:25:19 PM Modified: No Proc Algorithm: Intelliuan Min. Peak Height: 0.00 Cf Min. Peak width: 0.00 s< Smoothing Width: 3 pot RT Window: 30 0 st Expected RT: 14.9 ml Use Relative RT: No 2.47e+005 cps I Sample Name- *g120307a028* Sample 10: 'LCS-120306-1' File- *g120307a.wrt* Peak Name- *13C3PFHS-IS(IS)' Mass(es): '402.0/80.0 amu* Comment: *0.2ppbLCS* Annotation. ** Saammppllee Type: '' QC 0.937 ng/ml Calculated C< N/A Acq. Date: 3/8/2012 Acq. Time: 2.21:34 At Modified: Proc. Algorithm: Min. Peak Height: Min. Peak Width: Smoothing Width: RT Windowixpected P Use Rel : RT: 2.56e*005 cps Data printed by STW Printing Time: 12:30:11 PM Printing Date: Wednesday, April 11, 2012 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Results Name: gl20307a.rdb Sample Name *g120307a027* Sample 10. ' Peak Name: *13C3PFHS4S(IS) Mass(es): Comment 'Method Blank* Annotation: ** 27 Sample Type: Unknown Concentra ion: 0.945 Calculate Cone : H/A Acq. Date 3/8/2012 Acq. Time 2:00:24 Modi fled: No Proc. Alg rithm: IntelliCuan - IQA Min. Peak Height Min. Peak Width. 0.00 sec Smoothing Width- 3 RT Window 30.0 Expected 14.9 rain Use Relat ve RT. Ho Int. Type Valley Retention Time: 14.9 min 5 7308 counts Height: 70e+005 cps Start Tim 14.7 End Time: 15.2 min I Sample Name: *g120307a031* Sample ID: "LCS-120306-4* Fie. *g120307a.wtT Peak Name. *13C3PFHS4S(IS)* Mass(es)' *402.0/80.0 amu* Comment `2ppblCS' Annotation:** Sample Index: 31 Acq. Time: 3:25:11 > Modified: No Proc. Algorithm: IntelliQi Min. Peak Height: 0.00 Min. Peak Width: 0.00 Smoothing Width: 3 RT Window: 30.0 Expected RT: 14.9 Use Relative RT: Ho Height: Start Time: End Time: Valley > 14.9 866432 cc 2.47e*005 14.7 15.2 Page 1 of 12 Page 43 of 77 Ginger AG01330509 Printing Time: 12:30:11 PM Printing Date: Wednesday, April 11, 2012 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Results Name gl20307a.rdb Page 44 of 77 *** Ginger AG01330509 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Results Name gl20307a.rdb Data printed by STW Printing Time 12:30:11 PM Printing Date Wednesday, April 11 2012 Page 3 of 12 Page 45 of 77 Ginger AG01330509 Printing Time: 12:30:11 PM Printing Date: Wednesday, April 11, 2012 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Results Name: gl20307a.rdb Page 46 of 77 Ginger AG01330509 1) Sample Name: *g120307a034* Sample IO T.CS-120306-7* Fie- "g120307a.wifr I PeakName *13C8PFOS-tS{IS)* Mass(es) 507.0/80.0 amu I I Comment. *20ppb LCS* Annotation. " 34 Sample Type: QC 0.953 N/A ng/nL 9.0*4 Acq. Date: 3/8/2012 Acq. Tine: 4-28:40 AM 8.5*4 Modified. e s Min. Peak Height Min. Peak Width: Smoothing Width: RT Window: Expected RT: Use Relative RT: 0.00 0.00 3 30.0 15.` No cps sec 8.0*4 7.5*4 7.0*4 6.5*4 Int. Type: Retention Tit Height: Start Time End Time: Valley 15.8 297614 c< 9.20e*004 15.7 6.0*4 5.5*4 5.0*4 4.5*4 I 1 4.0*4 15.83 5000.0 13.5 ^ ___^ _1339 ^ I Sample Name ^120307*042* Sample 10: *GIP1 W11-02-27-026* Paak Nam '13C6PFOS-IS<IS)' Masses). *507.0/80.0 amu* Comment: 'JPAL-SW-TRIP1-LS* Annotation: ** SSaammppllee TIynpdee:x: 42 Concentration: Calculated Cone- Acq. Date : Acq. Time. 14.0 14.5 _ ^1388 1438 Fie- *g120307a.wifT Proc. Algorithm: Min. Peak Height Min. Peak Width: Smoothing Width: RT Window: Expected RT. Use Relative RT: Int. TypeRetention Time: Height. Start Time: End Time: 15.0 ^1488 15.5 1537 16.0 16.5 17.0 ____1587 1636 1686 17.5 1735 18.0 1785 Data printed by STW Printing Time: 12:30:11 PM Printing Date: Wednesday, April 11, 2012 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Results Name: gl20307a.rdb Page 5 of 12 Page 47 of 77 Ginger AG01330509 Printing Time: 12:30:12 PM Printing Date: Wednesday, April 11, 2012 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Results Name: gl20307a.rdb I Sample Name. 'g120307a053* Semple ID 'G Peak Name. *13C8PFOS-IS(l$)' Masses) ' Comment ' JPAL-SW-ET02-0" Annotation: " Sample Index: 53 Sample Type : Unknown Concentration: 0.960 ng/raL Calculated Cone: tl/A Acq. Date: 3/8/2012 Acq. line: 11:11:10 AM Modified: Ye* Proc. Algorithm: IntelliQuan - IQA . Peak Height: 0 00 cps . Peak Width: 0.00 sec Smoothing Width: 3 pointa RT Window: 30.0 sec >ected RT: 15.9 min : Relative RT: Mo Ini Type: Valle; Height: Start Tit End Time: 2-27-009' File 'g1203078.wrfP ISample Name' 'g120307a066' Sample ID ' GLP10-01-02-27-017* File' 'g120307a.wtfT Peak Name.'13C8PFOS-IS(IS)' Mass(es) *507.0/80.0 Comment ' JPAL-SW-ET04-0* Annotation. " Modified: Ye* Proc. Algorithm: IntelliQuan - IQA . Peak Height: 0.00 cps . Peak width: 0.00 sec Smoothing Width 3 points Page 6 of 12 1537 1587 Page 48 of 77 *** Ginger AG01330509 I Sample Name: *g120307a014* Sample ID. ` 1 I Peak Name: *PFBS* Masses) *299.0/99.0 ar U Comment: *0.25 ng/mL FC std In Synth. Water* Sample Index: 14 Sample Type: Standard 0.250 Calculated Cone: 0.285 Acq. Date: 3/7/2012 Acq. Time- 9:25:19 M Modi fled: No Proc. Algorithm: IntelliQ an - MQII Noise Percentage: 50 Base. Sub. Windov . 1.0 min Peak-Split. Facto r- 1 Report Largest Peek: Ho Min. Peak Height: M m . Peak width: 0. 0 sec Smoothing Width: 3 point RT Window: 30.0 sec Expected RT. 13.4 min Use Relative RT: Yes Int. Type- Area: Height: Start Time: End Time: Valley 13.4 433726 c 1.17e*00S 13.7 min cps min nun I Sample Name- *g120307a028` Sample 10' 1CS-120306-1* Fle: *g120307a.wifT Peak Name- *PFBS* Maares): ` 299.0/99.0 amu.299.0/80.0 amu* Cocnment: *0.2ppb LOS* Annotation: ** Sample Index: 28 Sample Type: QC IntelliQuan - MQII . Win Split. Factor: : Largest peak: ?eak Height: ?eak Width, ling Width: Expected RT: Use Relative Int. Type: Valley ;: 13.4 325226 cc 9.26**004 13.3 13.6 Data printed by STW Printing Time: 12:30:12 PM Printing Date: Wednesday, April 11, GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Results Name: gl20307a.rdb Sample Name. *g120307a027* Sample IO: *12001-160* Fie- *g120307a.wifT Peak Name. *PFB$* Mass(es): *299.0/99.0 amu,299.0/80.0 amu* Comment "Method Blank* Annotation: ** iample Type: Sample Index: 27 Proc. Algorithm: IntelliQuan - MQII Noise Percentage: Sub. .ndow: Peak-Split. Factor: Peport Largest Peak: Min. Peak Height: . Kin. Peak Width: Smoothing width: RT Window: 1 Expected RT: Use Relative RT: Int. Type: Vi I Sample Name: *g120307a031* Sample ID' ` LCS-120306-4' Peak Name. *PFBS* Mass(es): *299.0/99.0 amu,299......... Comment *2ppb LCS* Annotation' " Sample Index: 31 Sample Type: QC Concentration: 1.98 ng/raL Calculated Cone: 2.17 ng/mL Acq. Date: 3/B/2012 Acq. Time: 3.25:11 AM Proc. Algor: IntelliQuan Noisi Pert Window 50 Peak-Split. Facto. Report Largest Pe. No Min. Peak Height: U Min. Peak Width: 0 Smoothing width: 3 RT Window: 30.0 Expected RT: 13.4 Use Relative RT: Yes Type: Valley le: 13.4 3203368 coi 8.72e-*005 13.3 Page 7 of 12 Page 49 of 77 Ginger AG01330509 Data printed by STW Printing Time: 12:30:12 PM Printing Date: Wednesday, April 11, 2012 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Results Name: gl20307a.rdb Page 50 of 77 Ginger AG01330509 Printing Time: 12:30:12 PM Printing Date: Wednesday, April 11, 2012 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Results Name: gl20307a.rdb ISample Name -g120307a053* Sample IO. *GLP1CK>1-02-27-009* Peak Name *PFBS* Masses): *299.0/99.0 amu,299.0/80.0 amu* Sample Type: Concentration: tiled: t: Proc. Algorithm: Inte se Percentage- e. Sub. Window: Peak-Split. Factor: Report Largest Peak: Peak Height: . Peak width: Smoothing Width: RT Window: 30 Valley 13.3 47359 ce [Sample Name: 'g120307a066* Sample 10: 'GLP10-01-02-27-017* Peak Name: *PFBS` Mass(es). *299.0/99.0 amu,299.0/80.0 amu* Comment 'JPAL-SW-ET04-(r Annotation: " ample Index: 6 File: *g120307a.wHT IlfIt No . Algorithm: IntelliQui s Percentage: 50 Base. Sub. Window: 1.00 Peak-Split. Factor: 1 Report Largest Peak: No Peak Height: 0.0( Peak Width: 0.0( Smoothing Width: 3 iindow: 30 0 Expec 1 RT: Ose Relativ RT: . Type: Page 9 of 12 Page 51 of 77 *** Ginger AG01330509 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 ' Results Name: gl20307a.rdb Printing Time: 12:30:13 PM Printing Date: Wednesday, April 11 2012 Page 52 of 77 *** Ginger AG01330509 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Results Name gl20307a.rdb Printing Date: Wednesday, April 11, 2012 Page 53 of 77 Ginger AG01330509 I Sample Name: *g12O307aOS8* Sample ID: *GLP10-01-02-27-013* Peak Name. *PFHS` Masses): *399.0/99.0 amu,399.0/80.0 armi* Comment. `JPAL-SW-ET03-0* Annotation " Sample TIynpdee-x: Un5k8nown Concentration: N/A Calculated Cone: 0.0248 ng/raL Acq. Date: 3/8/2012 Acq. Time: 12:57:11 PM Fie' *g120307a.wifT Modified: RT Window: Expected RTUse Relative RT: fes 43373 counts 1.23e004 cps 14. min 14.9 min Time, mm Data printed by STW Printing Time: 12:30:13 PM Printing Date: Wednesday, April 11, 2012 T1 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Results Name gl20307a.rdb Page 12 of 12 Page 54 of 77 A tta c h m e n t C : A n a ly t ic a l M e th o d (s ) GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Page 55 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 3M Environm ental 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 t / ? / 'a Approved By: William K. Reagen, Technical Director, Environm ental Laboratory J /J C /i/ // Date ETS-8-044.1 P agel o f 22 Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis Page 56 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 1 Scope and A pplication This method describes the direct injection analysis of perfluorinated com pounds (PFCs) from w ater matrices using high-performance liquid chrom atography tandem mass spectrometry (HPLC/M S/M S). The m ethod is generally applicable but not limited to the m easurem ent o f perfluoroalkyl sulfonamides and perfluorinated alkyl acids (PFAAs) such as perfluorosulfonic acids (PFSAs) and perfluorocarboxylic acids (PFCAs) (Table 1). W a te r sam ples containing heavy particulate m ay require preparation by an alternate method such as ETS-8-154 " Determ ination o f Perfluorinated Acids, Alcohols, A m ides, and Sulfonates In W a te r By Solid Phase Extraction and High Performance Liquid Chrom atography/M ass Spectrom etry" . 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) A n a ly te Perfluorobutanoic acid Perfluoropentanoic acid Perfluorohexanoic acid Perfluoroheptanoic acid Perfluorooctanolc acid Perfluorononanoic acid Perfluorodecanoic acid Perfluoroundecanoic acid Perfluorododecanoic acid Perfluorotridecanoic acid Perfluorobutanesulfonic acid Perfluorohexanesulfonic acid Perfluorooctanesulfonlc acid Perfluorobutanesulfonamide Pefluorooctanesulfonamlde Chem ical A bstract 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 Minim um Reporting Level (M RL) is the Lim it o f Quantitation (LO Q) that m eets Data Quality Objectives (DQOs) that are developed based on the intended use of this method. Method Flexibility - This is a perform ance-based method and m ay be generally applied to the determ ination o f perfluorinated com pounds in w ater matrices w hen analysis batch quality control (QC) criteria are m et2. Each set o f sam ples are prepared in an analysis batch with calibration standards, LCSs, blanks, and continuing calibration check standards analyzed on the sam e instrument during a tim e period that begins and ends with the analysis o f the appropriate continuing calibration check standards. The laboratory is perm itted 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 m eet data quality objectives fo r the study. In all cases w here m ethod m odifications are im plem ented, the batch 1The method is supported by validation with internal standard calibration for C4-C13 PFCAs, C4, C6, and C8 PFSAs, and C8 perfluoroalkane sulfonamide in laboratory control samples under 3M method validation E l 1-0667. 2Guidance for establishing method QC Criteria based on a.) FDA May 2001, "Guidance for Industry, Bioanalytical Method Validation", b.) EPA Method 537, and c.) European Commission: Guidance for Generating and Reporting Methods o f Analysis in Support o f Pre-registration Data Requirements for Annex II (Part A, section 4) and Annex III (Part A,section 5) o f Directive 91/414, SANCO/3029/99 rev. 4 (11/07/00). ETS-8-044.1 Page 2 of 22 Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis Page 57 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 analytical Q C s (section 9) m ust be completed and pass Q C acceptance criteria (section 13) if the data from the analytical batch are to be reported. 2 M ethod Sum m ary W ater samples are analyzed as neat aqueous sam ple o r as solvent diluted aqueous samples by direct injection using LC/MS/MS. Sam ples containing heavy particulate m ay 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 sam ple is m ixed well prior to removing an aliquot o r diluting, if necessary, with A STM 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 com pounds (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 m ay be performed by external standard calibration. This is a perform ance-based method. Method uncertainty fo r each target analyte is determined fo r each analytical batch using multiple laboratory control spikes at multiple concentrations. This method also requires that the precision and accuracy for each sample be determ ined using field m atrix spikes to verify that the method is applicable to each sam ple matrix. Calibration standards for PFUnA, PFDoA, PFTrDA, and FOSA have been found to be unstable fo r m ore than 2 days in 100% water. Sam ples requiring analysis fo r these com pounds by this m ethod should be diluted 1:1 with m ethanol and analyzed against a calibration curve prepared in 1:1 synthetic groundw ater:M eO H. 3 D efinitions 3.1 A n a ly s is B a tch A set o f study samples that are prepared with calibration standards, laboratory control samples, and procedural blanks, and analyzed on the sam e instrument during a tim e period that begins and ends with the analysis o f the appropriate continuing calibration check standards. 3.2 A n a lytica l S am ple A portion o f a laboratory sample prepared for analysis. 3.3 C a lib ra tio n S tandard A solution prepared by spiking a known volum e o f the W orking Standard (W S) into a predeterm ined am ount o f A STM T ype I, HPLC grade water, o r other suitable w ater (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 L a b orato ry D uplicate Sam ple (LDS, o r Lab Dup) A laboratory duplicate sam ple is a separate aliquot o f a sam ple taken in the analytical laboratory that is analyzed separately with identical procedures. Analysis of LDSs compared to that o f the first aliquot give a m easure o f 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 58 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 3.5 Field B lank (FB)/Trip B lank (TB) A S T M T ype I, HPLC grade water, or other suitable water, placed in a sam ple container in the laboratory and treated as a sam ple in all respects, including exposure to sampling site conditions, storage, preservation and all analytical procedures. The purpose o f the T B is to determ ine if test substances o r o ther interferences are present in the field environm ent. This sam ple is also referred to as a Trip Blank. 3.6 Field D uplicate Sam ple (FDS, Field Dup) A sam ple collected in duplicate at the sam e tim e from the sam e location as the sample. The FDS is handled under identical circum stances and treated exactly the sam e throughout field and laboratory procedures. Analysis o f the FDS compared to that o f the first sample gives a m easure o f the precision associated with sam ple collection, preservation and storage, as well as with laboratory procedures. 3.7 Field M atrix S pike (FMS) A sam ple to which known quantities o f the target analytes, ISs and SRSs are added to the sam ple bottle in the laboratory before the bottles are sent to the field fo r collection o f aqueous samples. A known, specific volum e o f sample m ust be added to the sample container w ithout rinsing. This m ay be accom plished by making a "fill to this level" line on the outside of the sam ple container. T he FM S is analyzed to ascertain if any m atrix effects, interferences, or stability issues m ay com plicate the interpretation o f the sam ple analysis. 3.8 T rip B lank M atrix S pike (TBMS) A n aliquot o f A S T M T ype I, H PLC grade water, o r other suitable water, to w hich known quantities o f the target analytes, ISs and SRSs are added in the laboratory prior to the shipm ent o f the collection bottles. The TBM S is analyzed exactly like a study sam ple to help determ ine if the m ethod is in control and w hether a loss o f analyte o r analytical bias could be attributed to sam ple holding time, sam ple storage and/or shipm ent issues. A low and high TBM S are appropriate when expected sample concentrations are not known or m ay vary. 3.9 Inte rna l S tand ard (IS) A com pound added to each study sample, calibration standard, laboratory control samples, and procedural blanks at a consistent level (typically around 1 ng/mL). T he internal standard(s) are stable isotope labeled versions o f the target analytes. The area count ratio of the target analyte to the internal standard is used fo r calibration. Surrogate ISs are applied when stable isotope ISs o f target analytes are unavailable. A surrogate IS is not necessarily a stable isotope labeled version o f the target analyte, but is treated as an internal standard fo r quantitation. 3.10 L a b orato ry C o n tro l S am ple (LCS) An aliquot of control matrix to which known quantities o f the target analytes, ISs and SRSs (when applicable) are added in the laboratory at the tim e w hen sam ples are aliquotted. A t least three levels (two levels fo r SRSs) in triplicate are included, one generally at the low end o f the calibration curve and one near the mid range and the upper end o f the curve. The LCSs are analyzed exactly like a laboratory sam ple to determine w hether the stability o f the standards. LCSs should be prepared each day samples are aliquoted. 3.11 L a b o ra to ry M a trix S p ike (LMS) A laboratory m atrix spike is an aliquot o f a sam ple to w hich known quantities o f target analytes, ISs and SRSs (when applicable) are added in the laboratory. T he LMS is analyzed exactly like a laboratory sample to determine whether the sample matrix contributes bias to the analytical results. T he endogenous concentrations o f the analytes in the sam ple matrix m ust be determ ined in a separate aliquot and the m easured values in the LM S 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 59 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 3.12 Lab oratory Sam ple A portion or aliquot o f a sam ple received from the field fo r testing. 3.13 L im it o f Q uantitatio n (LOQ) T he lower limit o f quantitation (LLO Q) fo r an analytical batch is the lowest concentration that can be reliably quantitated within the specified limits o f precision and accuracy. The LLOQ is generally selected as the lowest non-zero standard in the calibration curve that m eets method acceptance criteria. T he LLOQ fo r each target analyte is established fo r each analysis batch as the lowest calibration standard with area counts at least twice that o f the average area counts o f the procedural blanks. The upper limit o f quantitation (U LO Q ) fo r 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 m eets method acceptance criteria is defined as the ULOQ. 3.14 M ethod/P rocedural Blank A n aliquot o f control m atrix that is treated exactly like a laboratory sam ple including exposure to all glassware, equipm ent, solvents, and reagents that are used with other laboratory samples. T he method blank is used to determ ine if te st substances o r other interferences are present in the laboratory environm ent, the reagents, or the apparatus. 3.15 Sam ple A sam ple is an aliquot removed from a larger quantity of material intended to represent the original source material. 3.16 S to ck S tandard S o lu tio n (SSS) A concentrated solution o f a single-analyte prepared in the laboratory with an assayed reference compound. 3.17 S urrogate Internal Standard A n IS that is not necessarily a stable isotopically labeled target analyte, but is treated as an internal standard fo r quantitation. Surrogate ISs are used when isotopically labeled counterparts o f the target analyte are not commercially or readily available. 3.18 S urrogate R ecovery Standard (SRS) A n isotopically labeled standard, not used as an internal standard, that is added to each sample and appropriate QC sample as a m eans to evaluate the method performance for a chemical class o f com pounds (e.g., PFSAs, PFCAs). 3.19 W o rkin g S tandard (WS) A solution o f several analytes prepared in the laboratory from SSSs and diluted as needed to prepare calibration standards and other required analyte solutions. 4 W arnings and C autions 4.1 H ealth and S a fety 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 w ear 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 60 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 T h e laboratory is responsible fo r maintaining a safe w ork environm ent and a current aw areness o f local regulations regarding the handling o f the chem icals used in this method. A reference file o f m aterial safety data sheets (M SDS) should be available to all personnel involved in these analyses. 4.2 C autions The analyst m ust be fam iliar with the laboratory equipm ent and potential hazards including, but not limited to, the use o f solvents, pressurized gas and solvent lines, high voltage, and vacuum systems. Refer to the appropriate equipm ent procedure or operator manual for additional information and cautions. 5 Interferences During sam ple preparation and analysis, m ajor potential contam inant sources are reagents and glassware. A ll materials used in the analyses shall be dem onstrated to be free from interferences under conditions o f 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 m ay include, but are not limited to: wash bottles, Teflon lined caps, autovial caps, HPLC parts, etc. T he use o f disposable m icropipettes o r pipettes to aliquot standard solutions is recom m ended to m ake calibration standards and matrix spikes. 6 Instrum entation, S upplies, and M aterials 6.1 In s tru m e n ta tio n Analytical balance capable of reading to 0.0001 g H PLC /M S/M S o r H PLC /M S system , as described in Section 10. 6.2 S upplies and M aterials Sam ple collection bottles-- HDPE (e.g., N algeneTM ) wide-m outh bottles with screw cap. N o te : Do not use fluorinated o r Teflon bottles o r lined caps. Coolers or boxes for sample shipment. 15-mL and 50-mL disposable polypropylene centrifuge tubes. Class A pipettes and volum etric flasks, various. 2 mL HPLC autovials Disposable pipettes, polypropylene or glass as appropriate Centrifuge capable of spinning 15-mL and 50-m L polypropylene tubes at 3000 rpm. 7 Reagents and Standards N ote: Suppliers and catalog num bers are for illustrative purposes only. Equivalent performance m ay be achieved using chemicals obtained from other suppliers. Do not use a lesser grade o f chem ical than those listed. 7.1 C h e m ic a ls W ater - Milli-Q, HPLC grade, or other suitably appropriate sources Calcium Acetate - A.C.S. Reagent Grade ETS-8-044.1 Page 6 of 22 Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis Page 61 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Magnesium Acetate - A.C.S. Reagent Grade Methanol - HPLC grade Amm onium Acetate - A.C.S. Reagent Grade 7.2 R epresentative Target A nalytes, 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, Am m onium perfluorooctanoate, (C8 Perfluorinated Acid) PFNA, Heptadecafluorononanoic Acid, (C9 Perfluorinated Acid) PFDA, Nonadecafluorodecanoic Acid (C 10 Perfluorinated Acid) PFUnA, Perfluoroundecanoic Acid, (C n Perfluorinated Acid) PFDoA, P erfluorododecanoic Acid, (C 12 Perfluorinated Acid) PFTrDA, Perfluorotridecanoic Acid, (C 13 Perfluorinated Acid) FBSA, Perfluorobutanesulfonamide FOSA, Perfluorooctanesulfonylamide PFBS, Potassium Perfluorobutanesulfonate PFHS, Perfluorohexanesulfonate PFOS, Potassium perfluorooctanesulfonate PFO A [1,2, 3 , 4 -13C], 13C4-isotopically labeled perfluorooctanoic acid (SRS) PFOS [1,2, 3 , 4 -13C], 13C4-isotopically labeled Perfluorooctanesulfonate (SRS) PFU nA [1 ,2-13C], 13C2-isotopically labeled Perfluoroundecanoic acid (SRS) A custom m ix o f ISs in a m ethanolic solution containing ([1 ,2,3,4-13C4]PFBA, [1,2 13C2]P FH xA, [1,2,3,4,5,6,7,8-13C8]PFOA, [1,2,3,4,5,6,7 8,9-13C9]PFNA, [1,2 - C JPFD A, [1,2,3,4,5,6,7 -13C7]PFUnA, [1,2 -^ C J P F D o A , [1,2,3 - C 3]PFHS, [1 ,2,3,4,5,6,7,8-13C 8]PFOS, and [1,2,3,4,5,6,7,8-13C8]P F O S A (W ellington Laboratories, Guelph, ON) in com bination with added ([1,2,3,4,5-13C5]PFPeA, ([1,2,3,4- C4]PFHpA, and [^ O J P F B S 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 R eagent P reparation 2 mM Am m onium acetate solution (Analysis)-- W eigh 0.3 g of Am m onium acetate and dissolve in 2.0 L o f reagent water. Synthetic G roundw ater (containing 25 ppm C a and Mg) - W eigh 0.61 g o f Calcium A cetate and 0.92 g o f Magnesium Acetate and dissolve in 6.0 L o f reagent water. Note: Alternative volum es m ay 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 62 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 7.4 S tock S tandard S o lution (SSS) and W orking Standard S o lution P re p a ra tio n The following standard preparation procedure serves as an example. W eighed am ounts and final volum es m ay be changed to suit the needs of a particular study. For example, pL volum es m ay be spiked into volum etric flasks when diluting stock solutions to appropriate levels. 100 pg/mL target analyte SSSs-- W eigh out 10 mg o f analytical standard (corrected fo r percent salt, acid [ETS-4-031] and purity) and dilute to 100 m L with methanol or other suitable solvent, in a 100 m L volum etric flask. Transfer to a 125 m L LDPE bottle o r other suitable container. Prepare a separate solution for each analyte. Expiration dates and storage conditions o f stock solutions should be assigned in accordance with laboratory standard operating procedure. A n exam ple o f purity and salt correction is given below fo r PFOS. m olecular w eight o f anion salt correction factor m oclecular w eight o f salt 499 P FO S (K + )sa lt co rre ctio n fa c to r = ------ = 0.9 2 7 5 538 10 mg C8F17S03'K + with purity 90% = 8.35 m g C8F17S03- (10 m g*0.90*0.9275=8.35 mg) 10 pg/mL (10,000 ng/mL) mixed working standard-- Add 5.0 m L each of the 100 pg/m L SSSs to a 50 m L volum etric flask and bring up to volum e with solvent. 1 pg/mL (1,000 ng/mL) mixed working standard-- Add 0.5 m L o f the 100 pg/m L SSSs to a 50 m L volum etric flask and bring up to volum e with solvent. 0.1 pg/mL (100 ng/mL) mixed standard-- A dd 0.05 m L o f the 100 pg/m L SSSs to a 50 m L volum etric flask and bring up to volum e with solvent. Storage Conditions-- Store all SSSs and w orking standards in accordance with laboratory standard operating procedure o r in a refrigerator at 42C fo r a m axim um period o f 6 m onths from the date of preparation. 7.5 C a libration S tandards Calibration can be perform ed by IS o r external calibration. Using the w orking standards described above, prepare calibration solutions in A S T M Type I water, HPLC water, other suitable water, o r a m ixture o f solvent and w ater using the information in Table 2 as a guideline. Note: Volum es of w ater or water/solvent mixtures and working standards may be adjusted to m eet 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 o f PFO A and PFOS, reference materials o f certified mixed linear and branched isom er are preferred. Alternately, reference materials o f primarily linear isomers of PFO A and/or PFOS m ay be used, however, w hen quantitating with predom inantly linear reference standards, additional LCS samples containing both linear and branched isomers of PFO A and PFO S are required3. 7.5.1 Internal Standard (IS) and Surrogate Recovery Standard (SRS) For IS calibration, stable isotope internal standards o f each target analyte or appropriate surrogate ISs should be spiked at the sam e level in all calibration standards. O nce the calibration standards have been prepared as stated above in Section 7.5, all calibration standards are spiked with a separate internal standard spiking solution. Typically the 3A report summarizing an assessment of the use of reference standards containing certified linear and branched isomers of PFOA/PFOS can be found in 3M report El 1-0560. ETS-8-044.1 Page 8 of 22 Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis Page 63 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 concentration o f the internal standard is consistent w ith th e internal standard concentration expected in the sam ples being prepared, usually 1 ng/mL. T he concentration o f the internal standard spiking solution is typically 2 pg/m L. A separate zero point o r m ethod blank is typically prepared at the sam e tim e as the calibration standards, using the sam e 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 sam e concentration as the calibration curve, typically at 1 ng/mL. If the sam ples being analzyed w ere pre-spiked with SRSs, the calibration curve prepared in Section 7.5 is spiked with a separate SRS spiking solution. Typically, the sam ple bottles are spiked with a SRS at 0.1 ng/m L. T he final calibration curve m ust consist o f at least six calibration points after analysis. The following table provides an exam ple o f spike concentrations and volum es used to achieve a multi-point extracted calibration curve with internal standard and surrogate standard. Table 1 lists recom m ended stable isotope internal standards fo r several PFSA and PFCA target com pounds. A custom m ix o f isotopically labeled target analytes in a methanolic solution containing ([1,2,3,4-13C4]PFBA, [1,2 - C JP F H xA , [1 ,2 ,3 ,4 ,5 ,6 7,8-13C8]PFOA, [1,2,3 ,4 ,5 ,6 ,7 ,8 ,9 -3C9]PFNA, [1,2,3,4,5,6 -13C6]PFDA, [1,2,3,4,5,6,7 -l3C7]PFUnA, [1,2 X J P F D o A , [1,2,3-13C 3]PFHS, [1,2,3,4,5,6,7,8-13C8]PFOS, and [1,2,3,4,5,6,7,8-13C8]FO SA (W ellington Laboratories, Guelph, ON) in com bination with added ([1,2,3,4,5-13C5]PFPeA, ([1,2,3,4-13C4]PFHpA, and [1802]PFBS can be used to prepare a stock IS solution. Alternative sources of certified stable isotope labeled target analytes are applicable. Alternatively, individual stable isotope ISs can be used to prepare a stock IS mixture. The table below lists the recom m ended stable isotope ISs and SRSs applied in the method. O ther stable isotope ISs and SRSs o f target analytes not listed in the table m ay be used if supported by validation and/or analysis batch Q C s meeting method acceptance criteria (e.g., [13C 2]-PFO A). The sam e internal standard should be used fo r a given analyte throughout the entire project/study. Note: som e o f the com pounds listed below are appropriate to use as surrogate ISs when a stable isotope IS o f 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 exam ples of spike concentrations and volum es 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 64 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Table 1. Stable Isotope PFCAs and PFSAs used fo r ISs and SRSs Compound Name Synonym or Acronym 13C4-Perfluorobutanoic acid [1,2,3,4-13C4]PFBA i3C4-Perfluoropentanoic acid [l,2,3,4,5-l3Cs)PFPeA 13C2-Perfluorohexanoic acid [1,2 -13C2]PFHxA 13C4-Perfluoroheptanoic acid [1,2,3,4-13C4]PFHpA l3C8-Perfluorooctanoic acid [1,2,3,4,5,6,7,8-13Cg]PFOA l3C9-Perfluorononanoic acid [1,2,3,4,5,6,7,8,9-I3C9]PFNA 13C6-Perfluorodecanoic acid [1,2,3,4,5,6 -13C6]PFDA 13C7-Perfluoroundecanoic acid [1,2,3,4,5,6,7 -13C7]PFUnA 13C2-Perfluorododecanoic acid [1,2 -l3C2]PFDoA l802-Ammonium Perfluorobutane sulfonate [180 2]PFBS l3C3-Ammonium Perfluorohexane sulfonate [1,2,3-13C3]PFHS 13Cg-Sodium Perfluorooctane sulfonate [l,2,3,4,5,6,7,8-13Cg]PFOS 13C8-Perfluorooctanesulfonamide l3C4-Perfluorooctanoic acid [l,2,3,4,5,6,7,8-13Cg]FOSA [1,2,3,4-i3C4]PFOA Analytical Purpose IS for PFBA IS for PFPeA IS for PFHxA IS for PFHpA IS for PFOA and [1,2,3,4 13C4]PFOA IS for PFNA IS forPFDA IS for PFUnA IS for PFDoA, *PFTA IS for PFBS IS for PFHS IS for PFOS and PFOS[l,2,3,4 13C4], IS for FOSA SRS for all PFCAs: C4-C8 Reference Standard Source Wellington Labs (Mix or Individual) Wellington Labs (Mix or Individual) Wellington Labs (Mix or Individual) Wellington Labs (Mix or Individual) Wellington Labs (Mix or Individual) Wellington Labs (Mix or Individual) Wellington Labs (Mix or Individual) Wellington Labs (Mix or Individual) Wellington Labs (Mix or Individual) RTI International (Individual) Wellington Labs (Mix or Individual) Wellington Labs (Mix or Individual) ' Wellington Labs (mix) RTI International (Individual) Wellington l3C2-Perfluoroundecanoic acid l3Cg-Perfluorooctane sulfonate [1,2 -13C2]PFUnA [l,2,3,4-13C4]PFOS SRS for all PFCAs C9-C13 Wellington SRS for all PFSAs: C4, C6, and C8 Wellington ETS-8-044.1 Page 10 of 22 Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis Page 65 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Table 2. Example Preparation o f Calibration Curve w ith ISs and SRSs Sample Description 0.025 ng/mL curve point 0.030 ng/mL curve point 0.04 ng/mL curve point 0.05 ng/mL curve point 0.1 ng/mL curve point 0.25 ng/mL curve point 0.5 ng/mL curve point 1 ng/mL curve point 2.5 ng/mL curve point 5.0 ng/mL curve point 10.0 ng/mL curve point 25.0 ng/mL curve point 50.0 ng/mL curve point 75.0 ng/mL curve point 100 ng/mL curve point Concentration o f WS, pg/mL 0.10 0.10 0.10 0.10 0.10 0.10 1.0 1.0 10.0 10.0 10.0 10.0 10.0 10.0 10.0 Volume o f Volume o f IS WS, pL (2 pg/mL), pL 25 50 30 50 40 50 50 50 100 50 250 50 50 50 100 50 25 50 50 50 100 50 250 50 500 50 750 50 1000 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 o f Surrogate, pL 12.5 15 20 25 50 125 250 500 25 50 100 NA NA NA NA Volume ofASTM Type 1Water, o r other suitable s o lv e n t(v, mL 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 N/A - Not Applicable (1) Samples requiring analysis for PFUnA, PFDoA, PFTrDA, and FOSA should be analyzed against a calibration curve prepared in 1:1 synthetic groundwater:MeOH. ETS-8-044.1 P age 11 o f 22 M ethod o f A nalysis fo r th e D eterm ination o f Perfluorinated C om pounds in W a te r by LC /M S/M S; D irect Injection A nalysis Page 66 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 8 Sam ple C ollection and B ottle Preparation Sample collection bottles are prepared by 3M Environmental Laboratory (or subcontract supplier) personnel for shipm ent at am bient tem perature to the collection site. Typically, four separate collection bottles are associated with a single collection site: sample, field duplicate sample, low field m atrix spike, and high field m atrix spike. Alternatively, the sam ple and field duplicate sam ple m ay contain S R S s in lieu o f additional target analyte low field m atrix spike and target analyte high field matrix spike samples. Depending on the scope of the project, additional replicates o f the field sam ple and field m atrix spikes m ay be added. Also, it is not uncomm on for additional mid-level field matrix spikes to be collected if the expected sam ple 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. (Volum es o f the bottles m ay vary depending on how m uch sam ple is required to m eet data quality objectives.) Sample collection volum es are project specific and based on data quality objectives. The Nalgene bottles do not require any pretreatm ent prior to use. Typically, placem ent o f a sam ple bottle volum etric "fill to here" line is done by using a sam ple bottle m arker template. Alternatively, bottles m ay 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 sam ple duplicate, low field matrix spike, high field matrix spike, sample/SRS field matrix spike, field duplicate sam ple/SRS field matrix spike, trip blank, o r trip blank m atrix spike. If each location has different designated spike levels, the label should also clearly indicate the sam ple location designation. Generally, a set o f bottles for a given collection site are prepared then grouped together in plastic bags fo r organizational purposes. F or each sam ple collection event, at least one set o f trip blank and trip blank matrix spikes are prepared. Bottle preparation should be docum ented in a N ote to File o r on a sam ple preparation w orksheet and should include the following information: date prepared, total num ber o f bottles prepared, num ber of sam ple sites, the standard identification num bers and spike volum es used to prepare spiked bottles, the "fill to here" volum e, and any other pertinent information needed fo r reconstructibility o f the data. The Note to File will be included in the final data package for the project. Sam ples are collected in th e field and shipped to the laboratory at am bient tem perature. 8.1 F ie ld M a trix S p ik e S a m p le (FMS) Field m atrix spike sam ples are a requirem ent o f the method. A FM S sam ple is defined as a QC sam ple to which known quantities o f appropriate target analytes are added to the sam ple bottle in the field o r in the laboratory before the bottles are sent to the field. The sam ple and field duplicate sam ple m ay Contain appropriate SRSs in lieu o f target analyte FM S samples. S am ple quantities are determ ined volum etrically o r gravimetrically. A known, specific volum e o r w eight o f sam ple is added to the sam ple container w ithout rinsing. Volum etric sam ple m easurem ents m ay be acquired by a laboratory applied "fill to this level" line on the outside o f the sample container. Target analyte FMS sam ples should be spiked at approximately 0.5-10 times the expected analyte concentration in the sample. If the expected range o f analyte concentrations is unknown, multiple spikes at varying levels m ay 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 fo r each sam pling location. In those instances w here SRSs are to be used in lieu o f target analyte FMS samples, the sam ple and field duplicate sam ple are spiked at approxim ately 2-5 tim es the target LOQ. T he FMS is analyzed to ascertain if m atrix effects or sam ple holding tim e contributes bias to the analytical results. For the sam ple bottles designated for m atrix spikes, an appropriate volum e o f m atrix spiking solution is added to th e em pty bottle prior to sampling. The volum e of spike solution added should produce the desired final concentration of target analytes once the bottle is filled with sam ple to the "fill to here line". The m atrix spiking solution(s) should be prepared in a suitable solvent and contain all o f the appropriate target analytes, ISs, and SRSs. The target analyte matrix spiking solution is often the sam e as the w orking standards used to create the calibration standards. An exam ple o f a bottle spike is given below. "Fill to here" volum e = 200 m L (A 250 m L 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 67 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 500 jj.L o f a 0.1 pg/m L spiking solution (containing the target analytes) is added to the bottle and the bottle cap prom ptly sealed. 8.2 Internal S tandard and S urrogate R ecovery Standard If analysis o f a surrogate recovery standard (SRS) Is included in the project objectives, an appropriate volum e o f a surrogate standard solution is added to all the bottles prior to sam pling and SPE. Typically sam ple bottles are spiked with surrogate recovery standards at a final desired spike concentration o f 0.1 ng/mL. If quantitation by internal standard (IS) is included in th e project objective, an appropriate volum e o f internal standard solution is added to all the bottles prior to sam pling and SPE. Typically sam ple bottles are spiked with internal standard at a final desired spike concentration of 1 ng/mL. For the trip blank, the S R S spike and IS spike is added to the bottle and then A S TM Type I w a te r (HPLC grade reagent w a te r o r o ther suitable w a te r m ay used) is added to the "fill to here" line. The bottle is capped and sealing tape m ay be placed around the outer edge o f the cap. Trip blank matrix spikes are prepared by adding the appropriate volum e o f target analyte spiking solution, IS, and SRS spiking solutions and filling the bottle to the desired volum e with the appropriate w ater and capping and sealing the cap. 9 Q uality C o ntrol and Data Q uality O bjectives 9.1 D ata Q u a lity O b je c tiv e s This method and required quality control sam ples is designed to generate data accurate to 30% with a targeted LOQ o f 0.025 ng/mL. A ny deviations from the quality control m easures spelled out below will be docum ented in the raw data and footnoted in the final report. 9.2 M ethod/P rocedural B lanks The m ethod/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 o f C C V samples, after every 10 sam ple injections, and at the end o f the analytical run. The m ean area count o r area ratios when using internal standard calibration, fo r each analyte in the method blanks m ust be less than 50% o f the area count counts o r area ratios when using internal standard calibration, o f the LOQ standard. The standard deviation of the area counts, or area ratios w hen using internal standard calibration, o f these method blanks should be calculated. A specific % R S D acceptance criteria is not specified but is assessed on an analytical batch basis. If the m ean area counts o r area ratios when using internal standard calibration, o f the method blanks exceed 50% o f the LOQ standard, then the LOQ m ust be raised to the first standard level in the curve that m eets criteria. Method blanks m ay be eliminated if technical justification can be provided (e.g. the procedural blank w as analyzed after an unexpectedly high level sample). If any procedural blanks are rem oved from the LOQ determination, docum ent in the raw data and report as appropriate. Laboratory Sam ple Replicates / Field Duplicate Sample Typically, sam ples are collected in duplicates in the field. T he relative percent difference (RPD) o f duplicate sam ples should be <20% fo r the precision o f sam ple 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 Lab o ra to ry M atrix S pikes (LMSs) LM Ss m ay be perform ed in lieu o f FM Ss if FM Ss have previously been perform ed fo r the sam ple matrix. Additionally, LM Ss m ay be perform ed in lieu o f FM Ss fo r a sam ple m atrix if the FMS levels w ere not appropriate fo r 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 sam ple and analyzed to determine the matrix effect on spike recovery efficiency o f each target analyte and appropriate SRSs. LM Ss should be prepared a t a m inim um o f one level and in duplicate. LMS concentrations should be prepared at approximately 0.5-10 tim es the endogenous concentration or approximately 4-10 tim es the LOQ concentration o f 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 68 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Lab matrix spike recoveries should fall within 30% of expected values. Sam ple data with LMS recovery outside o f 30% but within 50% o f the expected value are flagged and reported as outside o f QC acceptance criteria. Data with LMS recovery outside o f 50% o f the expected value are reported as NR, w here NR is defined as "Not Reportable" data outside o f QC acceptance criteria. 9.4 Lab C ontrol Sam ple Lab control spikes are prepared for each analysis batch to determine method accuracy and precision. LCSs should be prepared at three levels in triplicate fo r each target analyte and at a m inim um o f tw o levels in triplicate fo r appropriate SRSs. Low lab control spikes should be prepared at a concentration in the range of approximately four to ten tim es higher than the targeted lower LOQ, the mid lab control spikes should be prepared at a concentration near the mid-point o f 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 (m ethod precision) fo r each control spike level m ust be less than o r equal to 20% and the average recovery (method accuracy) for each control spike level m ust be 80-120%. Sam ple data for target analytes outside o f the laboratory control spike acceptance criteria will be handled as follows: . If the average recovery o f a spiking level falls outside m ethod acceptance, but at least 67% (6 out o f 9) o f LCS sam ples are within 20% o f their respective nominal value (33% o f the Q C samples, not all replicates at the sam e concentration, m ay be outside 20% o f 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 o f one of the spiking levels exceeded the analytical method uncertainty as determined by ETS-12012, that analytical batch uncertainty will be expanded fo r that particular study. If more than 67% o f the LCS sam ples fail to m eet method acceptance criteria, the data will not be reported. Calibration standards consisting o f mixed branched and linear isom er PFOS/PFOA are preferred. However, fo r PFO S/PFOA target analytes, if the calibration standards are comprised o f predominantly linear isomers only, at least one level o f triplicate LCSs should be prepared using PFOS/PFOA which contains a m ix of linear and branched isomers. These LCSs will be used to dem onstrate quantitative equivalency (or quantitative bias) of the isomeric mix when using a predom inantly linear standard fo r calibration. The mixed linear and branched isom er P FO S/PFO A LCSs recoveries should fall within 30% o f expected values. Alternatively, in lieu o f mixed branched and linear isom er PFOS/PFOA LCSs, mixed branched and linear isomer PFOS/PFOA TBM Ss m ay be applied to dem onstrate method accuracy and precision. 9.5 Field M atrix S pikes (FMSs) / S urrogate R ecovery Standards (SRSs) FMSs are prepared fo r each sampling location and analyzed to determ ine the matrix effect and sam ple holding tim e on the spike recovery o f each target analyte and/or appropriate SRS. Generally, each sam ple location represents a different sam ple and sam ple matrix. FMSs are QC samples to which known quantities o f 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 FM S are prepared fo r each sam pling location. T he sam ple and field duplicate sam ple m ay contain appropriate S R S s in lieu o f target analyte low field m atrix spike and target analyte high field m atrix spike samples. Field matrix spike method acceptance criteria are recoveries within 30% o f the expected value. If FMS recovery (target analyte o r SRS spike) is outside o f 30% o f the expected value o r could not be assessed because the FM S (target analyte) w as spiked at an inappropriate level, the sam ple result is reported as follows: 1. ) If target analyte FM S recovery could not be assessed because the FM S's w ere 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 w ere inappropriate. 2. ) If multiple target analyte FMS's w ere prepared on a sam ple and the closest FMS level to the reported sam ple meets the 30% acceptance criteria but additional FMS's are outside the 30% acceptance range, the data are reportable and flagged to indicate that w hile there w ere failing FM S 's, the uncertainty will not be expanded since the m ost 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 69 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 3. ) If the target analyte FMS recoveries are outside o f the 30% acceptance range but at least 30 acceptable historical reportable FMS sam ple results are available, the data m ay 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 FM S recovery is outside o f 50% , the sam ple result is reported as NR, w here 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 tim es the endogenous level to be used without justification to determ ine the statem ent o f accuracy fo r analytical results. Note: It is possible fo r bottles utilized fo r Field Matrix Spike sam ples to be under-filled o r over-filled during sam ple collection. Since this scenario will effect the actual concentration of the FMS sam ple (surrogate and internal standard concentrations will also be effected, if used), it is im portant that any obvious under-filling or over-filling o f sam ple bottles be docum ented in the data package and taken into account in the FMS, ISs, or SRSs recovery calculations. Sam ples 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 sam ple and the field duplicate should be used to calculate the recovery. 10 P rocedures 10.1 W a te r S am ple P re p a ra tio n This method is applicable to w ater samples. S am ples containing heavy particulate m ay not be suitable fo r analysis by this method. Sam ples containing suspended particulate should be centrifuge prior to removing a sam ple aliquot, o r filtered. Thoroughly m ix sam ple before removing an aliquot and placing in a labeled autovial. Dilute sample, if necessary, with A STM Type I water, HPLC water, other suitable water, or solvent (m ethanol). Lab control spikes are prepared for each analysis batch to determine method accuracy and precision. LCSs should be prepared at three levels in triplicate fo r each target analyte and at a m inim um o f tw o levels in triplicate fo r appropriate SRSs. Low lab control spikes should be prepared at a concentration in the range o f approxim ately fo u r to ten tim es higher than the targeted lower LOQ, the mid lab control spikes should be prepared at a concentration near the mid-point o f the calibration curve and the high lab control spikes at approxim ately 80% o f the upper LOQ. For IS quantitation, stable isotope internal standards o f each target analyte o r appropriate surrogate ISs should be spiked at the sam e level as the sam ples being analyzed, in all LCSs. If LCSs are being prepared using synthetic groundwater, allow the LCSs samples to equilibrate for a minimum o f 4 hours before aliquoting for analysis or diluting with solvent (methanol). 11 S a m p le A n a ly s is - LC /M S/M S 11.1 In s tru m e n t S e tu p N ote: In this example, an Applied Biosystem s Sciex API 4000 (API 5000 o r API 5500) Tandem M ass S pectrom eter (LC/M S/M S) is used. O ther brands/m odels o f LC /M S/M S instrum ents as well as single q u a d ru p le m ass spectrometers (LC/MS) may be used as long as the method acceptance criteria are met. Brand names, suppliers, part numbers, and models are fo r illustrative purposes only. Equivalent performance m ay be achieved using apparatus and materials other than those specified here, but demonstration of equivalent perform ance that m eets the requirem ents o f this m ethod is the responsibility o f the laboratory. T he operator m ust optimize and docum ent the equipm ent 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 70 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 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: T urbo Ion S pray (ABS) Mode: Electrospray Negative Scan Type: MRM (Multiple Reaction Monitoring) Computer: Dell DHM Software: W indow s 2000 or W indows 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: O ne or m ore C18 H PLC analytical colum ns (2.1 m m x 100 mm, 5pm o r 2.1 m m x 5 0 mm, 5(im) m a y b e attached on-line after the purge valve and before the sample injection port to retard and separate any residue contam inants that m ay be in the m obile phase and/or H PLC system. HPLC Column: Betasil C18, 4.6m m x 100mm, 5|om (Therm oElectron Corporation) Column Temperature: 35C Injection Volume: 5pL Mobile Phase (A): 2m M A m m onium A cetate in A S T M Type I w a te r (See 7.3) Mobile Phase (B): Methanol Table 3. Liquid Chrom atography G radient Program. Step Number 0 1 2 3 4 5 Total Time (min) Flow Rate (pL/min) 0 2.0 14.5 15.5 16.5 20.0 750 750 750 750 . 750 750 Percent A (2 mM ammonium acetate) 97.0 97.0 5.0 5.0 97.0 97.0 Percent B (Methanol) 3.0 3.0 95.0 95.0 3.0 3.0 Note: O ther HPLC gradients m ay be used as long as the method criteria and project data quality objectives are met. It m ay be necessary to adjust the H PLC gradient in o rder to optim ize instrum ent performance. C olum ns with different dim ensions (e.g. 2.1m m x 30m m ) and colum ns from different m anufacturers (Keystone Betasil C18 etc.) m ay 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 71 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 Table 4 Suggested MRM Transitions fo r Target Analytes, Surrogates, and Internal Standards Analyte PFBA (C4 Acid) PFPeA (C5 Acid) PFHxA (C6 Acid) PFHpA (C7 Acid) PFOA (C8 Acid) PFNA (C9 Acid) PFDA (CIO Acid) PFUnA (Cl 1Acid) PFDoA (Cl2 Acid) PFTA (C l3 Acid) FBSA (C4 Sulfonamide) FOSA (C8 Sulfonamide) PFBS (C4 Sulfonate) PFHS (C6 Sulfonate) PFOS (C8 Sulfonate) r1,2,3,4 -I3C41PFBA n,2,3,4,5 -13C5lPFPeA [1,2 -13C21PFHxA f1,2,3,4- 13G,lPFHpA [1,2,3,4,5,6,7,8-13C81PF0A [1,2,3,4,5,6,7,8,9-'3C91PFNA [1,2,3,4,5,6-13C61PFDA [1,2,3,4,5,6,7 -,3C7lPFUnA [1,2-'3C21PFDoA [180 21PFBS [1,2,3-13C31PFHS [1,2,3,4- 13C41PF0S [1,2,3,4,5,6,7,8-13Cs1FOSA [1,2,3,4-i3C41PFOA [1,2,3,4- 13C4lPFOS [1,2 -13C2lPFUnA Analyte Description Target Target Target Target Target Target Target Target Target Target Target Target Target Target Target IS for PFBA IS for PFPeA IS for PFHxA IS for PFHpA IS for PFOA IS for PFNA IS for PFDA IS for PFUnA IS for PFDoA and PFTA IS for PFBS IS for PFHS IS for PFOS IS for FOSA Surrogate (C4-C8 Acids) Surrogate(Sulfonates, FOSA) Surrogate (C9-C13 Acids) Mass Transition 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 03 (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 fo r m onitoring the analytes is an option. T h e use o f one daughter ion is acceptable if data sensitivity and selectivity is achieved and provided that retention tim e criteria are m et to assure adequate specificity. W hile the daughter ions m ay be chosen at th e discretion o f the analyst, m ass transition 99 is suggested fo r PFOS. Quantitation m ay be perform ed using the total ion chrom atogram (TIC, o r sum m ed M RMs) fo r a given analyte. For example, the PFO A TIC would sum all three o f the monitored transitions. Use o f the suggested prim ary ion is recom m ended. Retention tim es m ay vary slightly, on a day-to-day basis, depending on the batch o f m obile phase and the gradient, colum n, guard colum n(s) used etc. Drift in retention tim es 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 C a libration C urve Quantitation is by internal standard or external standard calibration. Calibration standards m ay be prepared in A S TM Type I, HPLC water, other suitable water, o r a solvent/w ater mixture. If internal standard calibration does not m eet 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 72 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 recom m ended application o f available internal standards. Quantitation o f P FO A and PFO S is by sum m ed analyte-specific m ass transitions. Analyze the standard curve prior to each set o f samples. If internal standards were added to the calibration standards area ratios are used to generate the calibration curve. The standard curve m ay be plotted using a linear regression (y = m x + b), w eighted 1/x o r unweighted, o r by quadratic fit (y = ax2 + bx + c), w eighted 1/x or unweighted, using suitable software. The mathematical method used to calculate the calibration curve should be applied consistently throughout a study. A n y change should be thoroughly docum ented in the raw data. High and/or low points m ay be excluded from the calibration curves to provide a better fit over the range appropriate to the data or because they did not m eet 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 tw ice that o f the average area counts (or area ratio if quantitating by IS) of method and/or solvent blanks. The coefficient o f determination (r2) value fo r the calibration curve m ust be greater than o r equal to 0.990 (or a correlation coefficient (r) o f 0.995). Each point in the curve m ust be within 25% o f the theoretical concentration with the exception o f the LLOQ, which m ay be within 30%. Justification for exclusion o f calibration curve points will be noted in th e raw data. A m inim um o f 6 points will be used to construct the calibration curve. If the calibration curve does not m eet acceptance criteria, perform routine maintenance o r prepare a new standard curve (if necessary) and reanalyze. 11.3 C o n tin u in g C a libration V e rifica tio n (CCV) Continuing calibration verifications (CCV) are analyzed to verify the accuracy o f the calibration curve. Analyze a mid-range calibration standard, one of the sam e standards used to construct the calibration curve, at a m inim um after every tenth sample, not including solvent blanks, with a m inim um o f one per sam ple set. Calibration verification injections m ust be within 25% to be considered acceptable. The calibration curve and the last passing C C V will then bracket acceptable samples. Multiple C C V levels m ay be used. Sam ples m ust be bracketed by passing CCVs o r the calibration curve and a passing C C V to be reportable. 11.4 System S u ita b ility A minimum o f three system suitability samples should be injected at the beginning of each analytical run, prior to the analysis o f the calibration curve. Typically these sam ples are at a concentration near the mid-level of the calibration curve and are repeated injections from one autosam pler vial. It is suggested that the system suitability injections have area counts or area ratios when using internal standard calibration, with a target RSD o f <5% and a target retention tim e RSD o f <2%. There is no defined acceptability limit on these results as the % R S D value is dependent on the num ber o f MRM transitions being m onitored in the LC /M S/M S run or tim e period. Ultimately, any effects on these param eters fo r the System Suitability sam ples will also be evident on all standards and QC sam ples analyzed as p a rt o f the analysis batch. A n y effect o f system suitability is incorporated within QC acceptance criteria.4 11.5 S am ple A n alysis and QCs For each analysis batch, the instrum ent analysis run sequence should include an initial calibration curve, samples, FDSs, interspersed blanks, interspersed CCVs, appropriate Q C s (i.e., LCSs, LMSs, FMSs, TBM Ss, and TBs), and a final C CV or calibration curve bracketing sam ples and appropriate QCs Inject the sam e volum e (between 5 - 1 0OpL) o f each standard, analytical sample and blank into the instrument (unless an on-instrum ent sam ple dilution is desired). Sam ples containing analytes that are quantitated above th e concentration o f the highest standard in the curve should be further diluted and reanalyzed. 4 3M Environmental Laboratory study E08-0096 evaluated the effect on these results as a function of the number of MRMs being monitored. ETS-8-044.1 Page 18 of 22 M ethod o f A nalysis fo r th e D eterm ination o f Perfluorinated C om pounds in W a te r by LC /M S /M S; D irect Injection A n a ly s is Page 73 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 12 Data A n a ly s is a n d C a lc u la tio n s _____________________________________________ T he chrom atography analysis software will typically calculate the am ount o f target analyte in the sam ple extracts using the established calibration curve. Calculate the percent recovery of the LCS using the following equation: LCS Concentration (-^ -) LCS% recovery = ------------------------- 2 * ^ - *100% Spike Concentration ( ^ - ) mL Calculate the percent recovery o f the LMS using the following equation: LMS % recovery na nq LMS C oncentration (-- ^ -) - C oncentration o f Sam ple (-- ^-) ___________________ mL _______________ mL nq Spike Concentration ( - 2 - ) mL 100% For samples fortified with known amounts of analyte prior to extraction, use the following equation to calculate the percent recovery. R ecovery = Total analyte found (ng/m L) - A ve ra g e analyte found in sam ple (ng/m L) x100 Analyte added (ng/m L) 13 A nalysis Batch M ethod Perform ance C riteria A ny method perform ance param eters that are not achieved m ust be considered in the evaluation o f the data. N onconform ance to any specified param eters m ust be described and discussed in the final report if the Technical M anager (non-GLP study) or Study Director (GLP study) chooses to report the data. If criteria listed in this m ethod perform ance section are not met, m aintenance m ay be perform ed on the system and sam ples reanalyzed, o r other actions taken as appropriate. D ocum ent all actions in the raw data. If data are to be reported when performance criteria have not been met, the data m ust be footnoted on tables and discussed in the te xt o f the report. 13.1 S ystem S u ita b ility - A n a ly s is B a tch A minimum of three system suitability samples should be injected at the beginning of each analytical run. These sam ples are run prior to the calibration curve. It is suggested that the system suitability injections have area counts with a target R SD o f <5% and a target retention tim e RSD o f <2%. T here is no defined acceptability limit on these results as the % RSDs are dependent on the num ber of MRM transitions being monitored in the LC /M S/M S run o r tim e period. A n y effect o f system suitability is incorporated in the QC acceptance criteria. 13.2 C a lib ra tio n and L im it o f Q u a n tita tio n (LOQ) - A n a lysis Batch Calibration Curve: The coefficient o f determ ination (r2) value fo r the calibration curve m ust be greater than or equal to 0.990 corresponding to a correlation coefficient (r) = 0.995. Each point in the curve m ust be within 25% o f the theoretical concentration with the exception o f 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 o f the calibration curve. T h e 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 74 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 o f each curve point m ust be within 25% o f the theoretical value (within 30% fo r lowest curve point). Samples that are bracketed by CCVs not meeting these criteria m ust be reanalyzed. Lim its o f Quantitation (LOQ): The lower LOQ (LLO Q) is the lowest non-zero active standard in the calibration curve; the peak area o f the LLOQ m ust be at least 2X that o f the average area counts fo r all prepared procedural blank(s). By definition, the m easured value o f the LLOQ m ust be within 30% o f the theoretical value. Demonstration o f Specificity: Specificity is dem onstrated by chrom atographic retention tim e (within 4% of standard) and the m ass spectral response o f unique ions. 13.3 B lanks - M ethod/P rocedural B lanks and T rip Method/Procedural Blanks: Multiple procedural blanks should be interspersed throughout th e analysis batch and the analytical sequence. A t a minimum, method blanks are analyzed prior to instrument calibration, prior to the analysis o f C C V samples, after every 10 sam ple injections, and at the end o f the analytical run. The m ean area counts (or area ratios w hen using IS calibration) fo r each analyte m ust be less than 50% o f the area count o f the LOQ standard. If the area counts of the procedural blanks exceed 50% o f the LOQ standard, then the LOQ m ust be raised to the first standard level that meets criteria. Trip Blank: A trip blank o f A STM Type I w ater (or lab equivalent) is prepared in a sam ple 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 fo r each analyte are included with the reported sam ple results. 13.4 Data A ccu ra cy and P recision - A n a lysis Batch Lab Control Spikes: The average recovery at each LCS level fo r 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 o f a spiking level falls outside m ethod acceptance, but at least 67% (6 out o f 9) o f LCS sam ples are within 20% o f their respective nominal value (33% o f the Q C samples, not all replicates at the sam e concentration, may be outside 20% of nominal value), the average recovery will be flagged as outside m ethod acceptance criteria. A ll LCS sam ples will be control charted as per ETS-12-012. If the average recovery o f one o f 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 isom er PFOA and PFOS should be within 70-130% and the percent relative standard deviation o f the recoveries m ust be less than or equal to 20%. Field Duplicates: The relative percent difference (RPD) o f duplicate sam ples should be less than 20% fo r the precision o f sam ple preparation and analysis to be considered in control. Replicate sam ples not m eeting the 20% RPD criteria are flagged and reported as outside of QC acceptance criteria. Field Matrix Spikes: FMS acceptance criteria are recoveries within 30% o f the expected value fo r each target analyte and appropriate SRS. Sam ple data with FMS recovery outside o f 30% but within 50% o f the expected value are flagged and reported as outside o f Q C acceptance criteria. Data with FMS recovery outside o f 50% o f the expected value are reported as NR, w here NR is defined as "N ot Reportable" data outside o f QC acceptance criteria. If FMS recovery could not be assessed because FMSs were at an inappropriate level, then Laboratory M atrix Spikes (LM Ss) m ay 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 A n a lytica l M ethod U ncertainty Analytical method uncertainty fo r each target analyte and SRS is determ ined with control charted historical analysis batch LCS data for the method and reported with each analysis batch.5 Uncertainty determinations 5 Method uncertainty based on INTERNATIONAL ANS/ISO/IED STANDARD 17025 reference (GUM, Guide to the Expression of Uncertainty in Measurement). Method application demonstrated in ETS-12-012, citing references: a.) EURACHEM/CITAC Guide, "Quantifying Uncertainty in Analytical Measurement," Second Edition; Editors: S.L.R. Ellison, M. Rosslein, and A. Williams. b.)Georgian, Thomas, "Estimation o f Laboratory Analytical Uncertainty Using Laboratory Control Samples," Environmental Testing & ETS-8-044.1 Page 20 of 22 Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis Page 75 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 are based on INTERNATIONAL ANS/ISO/IED STANDARD 17025 reference (GUM, Guide to the Expression o f Uncertainty in M easurem ent) and described in ETS-12-012. A t least thirty data points are required fo r determ ining analytical m ethod uncertainty. T he m ethod uncertainty is defined as 2x the standard deviation o f the percent recoveries o f the pooled lab control spikes. W hile all LCS data points are control charted, only the m ost recent fifty data points are used fo r determining the method uncertainty. W hen 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 m eet the 30% recovery criteria at a level appropriate to the endogenous level, and the LCS m eet the 20% recovery criteria, then the uncertainty o f the data is determ ined as within 10020%. Analysis batch sam ple 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 o f 50% o f the expected value are reported as NR, w here N R is defined as "N ot Reportable" data outside o f QC acceptance criteria. If FMS recovery could not be assessed because FMSs w ere at an inappropriate level, then Laboratory Matrix Spikes (LM Ss) m ay be substituted. If LM S 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 m eet the 30% recovery criteria, and historical FMS data does not exist, the analytical uncertainty is evaluated on a sam ple-by-sam ple basis, the data may be reported with expanded uncertainty and are flagged. 13.6 Q uantitatio n o f PFOA/PFOS - A n a lysis Batch Calibration standards consisting o f mixed branched and linear isom er PFO S/PFOA are preferred. Quantitation is perform ed by integrating the linear and branched isom ers together. Alternately, the linear and branched isomers can be integrated separately, applying the appropriate true value to each calibration curve point fo r each isomer. The LCS and sam ples 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 sum m ed to produce the final result. Integrating the linear and branched isomers separately m ay be helpful for those sam ples where the linear/branched ratios do not closely match those of the reference standards. However, fo r PFO S/PFOA target analytes, if the calibration standards are comprised o f predom inantly linear isomers only the method requires the addition o f LCSs o f mixed branched/linear isomer PFOS/PFOA. The purpose o f including these LCSs is to dem onstrate quantitative equivalency (or quantitative bias) o f the . isom eric m ix when using a predom inantly linear PFO S o r P FO A standard fo r calibration. Alternatively, in lieu o f mixed branched and linear isom er PFOS/PFOA LCSs, mixed branched and linear isom er PFOS/PFOA TBM Ss m ay be applied to demonstrate method accuracy and precision. A n alternate m ethod o f quantitation can be perform ed w hereby only the linear isom er o f P FO S /P FO A 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 sum m ed 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 th e a range o f the calibration curve that is com parable regardless of whether the calibration curve w as generated using predominantly linear isomers o f PFOS/PFOA or linear plus branched isomers of PFOS/PFOA. 14 P ollution Prevention and W aste M anagem ent W aste generated w hen performing this method will be disposed o f appropriately. The original samples will be archived at the 3M Environm ental Laboratory in accordance with internal procedures. Analysis, November/December 2000. c.)Taylor, B.N. and CE. Kuyatt, NIST Technical Note 1297,1994 Edition: "Guidelines for Evaluating and Expressing the Uncertainty o f NIST Measurement Results."d.)Adams, T.M., "A2LA Guide for the Estimation o f Measurement Uncertainty in Testing", July 2002. ETS-8-044.1 Page 21 of 22 Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis Page 76 of 77 GLP10-01-02; Interim Report 26 Analysis of PFBS, PFHS, and PFOS Bert Jeffries Landfill - February 2012 15 Records Each data package generated fo r a study m ust include all supporting information fo r reconstruction o f the data. Information fo r the data package m ust include, but is not limited to the following items: study o r project number, sample and standard prep sheets/records, instrument run log (instrum ent batch records, instrument acquisition method, sum m ary pages), instrument results files, chrom atograms, calibration curves, and data calculations. .. 16 A ffe cte d D ocum ents None. 17 R evisions R e v is io n Number 1 Summary of Changes Section 1. Included the use of internal standard calibration by this method. Section 2. Included the use of internal standard calibration by this method. Included the use of a solvent/water mixture when analyzing forPFUnA, PFDoA, PFTrDA, and FOSA. Section 3. Added definitions for internal standard, surrogate internal standard, and surrogate recovery standard. Section 6.Removed the details regarding the instrument parameters to section 10 of the method. Section 7. Updated reference standards to include internal standards and surrogates. Changed concentration levels for working standards and included the use of internal standards and surrogates. Section 8. Inserted a new section on sample bottle preparation. Section 9 Quality Control. This section was previously section 10 in ETS-8-044.0. Updated QC criteria to be consistent with method ETS-8-154.4. Section 10 Procedures. This section w as previously section 8 (Sample Handling) in ETS-8044.0. Added detail regarding the preparation of LCSs. Included the use of methanol as a dilution solvent. Section 11 Sample Analysis. This section was previously section 10 in ETS-8-044.0. Included the details regarding the instrument parameters. Section 12 Data Analysis and Calculations. This section was previously section 11 in ETS8-044.0. Removed the equation for calculating the analytes concentration, indicating that this is done by the instrument software. Section 13 Method Performance. This section was previously section 12 in ETS-8-044.0. Updated QC criteria to be consistent with ETS-8-154.4. Added information on the determination of analytical method uncertainty and quantitation of PFOA/PFOS. Section 14 Pollution Prevention. This section was previously section 13 in ETS-8-044.0. Section 15 Records. This section was previously section 14 in ETS-8-044.0. Section 16 Affected Documents. This section was previously section 15 in ETS-8-044.0. Section 17 Revisions. This section w as 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 77 of 77