Document jm4Y9pRww2eDjnj4yxp5585kZ

EOS-0210 Interim Report #18 Northern Alabama Potable Witter Systems, December 2005 Interim Report #18: Analysis of Water Samples from Northern Alabama Potable Water Systems, December 2005 Study Title A nalysis of Perfluo robutanesulfonate (PFBS), Perfluorohexanesulfonate (PFHS), an d Perfluorooctanesulfonate (PFOS) in Wate r , So il, S edim ent, Fis h , Clam s, V eg etatio n , Sm all Mam m al Liver an d Sm all Mam m al S erum Using LC/MS/MS FOR THE 3M DECATUR MONITORING PROGRAM Data Requirement EPA TSCA Good Laboratory Practice Standards 40 CFR 792 Author M ichelle D. Malinsky, Ph.D 3M Environm ental Laboratory Interim Report Completion Date Date o f signing Performing Laboratory 3M Environmental Laboratory Building 2-3E-09 935 Bush Ave. St Paul, MN 55106 Project Identification E05-0210 Total Number of Pages 122 The testing reported herein meet the requirements of ISO/IEC 17025-1999 `General Requirements for the Competence of Testing and Calibration Laboratories' , in accordance with the A2LA Certificate #2052-01. Testing that complies with this International Standard also operate in accordance with ISO 9001/ISO 9002 (1994). E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 1 of 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 This page has been reserved for specific country requirements. E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 2 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 GLP Compliance Statement Study Title: Analysis of Perfluorobutanesulfonate (PFBS), Perfluorohexanesulfonate (PFHS), and Perfluorooctanesulfonate (PFOS) in Water, Soil, Sediment, Fish, Clams, Vegetation, Small Mammal Liver and Small Mammal Serum Using LC/MS/MS for the 3M Decatur Monitoring Program Interim Study: Analysis of Water Samples from Northern Alabama Potable Water Systems, December 2005 Interim Study Identification Number: E05-0210 Interim Report #18 This study was conducted in compliance with Toxic Substances Control Act (TSCA) Good Laboratory Practice (GLP) Standards, 40 CFR 792, with the exceptions listed below: Exceptions to GLP compliance: None Sponsor Representative Jaisimha Kesari P. E., DEE, Study Director Date Certificate #2052-01 The testing reported herein meet the requirements of ISO/IEC17025-1999 `General Requirements for the Competence of Testing and Calibration Laboratories' , in accordance with the A2LA Certificate #2052-01. Testing that complies with this International Standard also operate in accordance with ISO 9001/ISO 9002 (1994). E05-0210 Interim Report 018 Northern Alabama Potable Water Systems, December 2005 Page 3 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December2005 Quality Assurance Statement Study Title: Analysis of Perfluorobutanesulfonate (PFBS), Perfluorohexanesulfonate (PFHS), and Perfluorooctanesulfonate (PFOS) in Water, Soil, Sediment, Fish, Clams, Vegetation, Small Mammal Liver and Small Mammal Serum Using LC/MS/MS for the 3M Decatur Monitoring Program Interim Study: Analysis of Water Samples from Northern Alabama Potable Water Systems, December 2005 Study Identification Number: E05-0210 Interim Report #18 This study was audited by the 3M Environmental Laboratory Quality Assurance Unit (QAU), as indicated in the following table. The findings were reported to the study director and laboratory management. Inspection Dates February 15,2006 Phase Data/Report Date Reported to Management Study Director March 17,2006 March 17,2006 Date E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December2005 Page 4 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December2005 Table of Contents GLP Compliance Statement.............................................................................................................. 3 Quality Assurance Statement............................................................................................................ 4 Table of Contents.............................................................................................................................. 5 List of Tables..................................................................................................................................... 6 Study Information.............................................................................................................................. 7 Summary and Introduction.................................................................................................................8 Test Samples...................................................................................................................................10 Reference Substances.....................................................................................................................10 Method Summaries..........................................................................................................................11 Sample Collection................................................................................................................ 11 Preparatory and Analytical Methods..................................................................................... 11 Extraction............................................................................................................................. 11 Analysis................................................................................................................................ 11 Analytical Results............................................................................................................................ 13 Calibration............................................................................................................................ 13 Limit of Quantitation (LOQ)................................................................................................... 13 Blanks.................................................................................................................................. 13 Solvent Blank........................................................................................................................13 Method Blank....................................................................................................................... 13 Trip Blank............................................................................................................................. 14 System Suitability................................................................................................................. 14 Continuing Calibration.......................................................................................................... 14 Lab Control Spikes (LCSs)................................................................................................... 14 Sample Duplicates............................................................................................................... 15 Field Matrix Spikes............................................................................................................... 15 Data Summary and Discussion........................................................................................................15 EOS-0210 Interim Report 018 Northern Alabama Potable Water Systems, December 2005 Page 5 o f 122 EOS-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Statistical Methods and Calculations................................................................................................ 19 Accuracy and Precision Equations........................................................................................ 19 Determination of Analytical Uncertainty.................................................................................19 Statement of Conclusion..................................................................................................................20 References..................................................................................................................................... 20 List of Attachments.......................................................................................................................... 20 Signature Page................................................................................................................................21 List of Tables T a b le ! Sample Results Summary................................................................................................... 9 Table 2. Sample Description Codes................................................................................................. 10 Table 3. Study Reference Substances............................................................................................. 10 Table 4. Sample Collection Information...........................................................................................11 Table 5. Instrument Parameters.......................................................................................................12 Table 6. Liquid Chromatography Gradient Program..........................................................................12 Table 7. Mass Transitions................................................................................................................12 Table 8. Method Blank Area Counts................................................................................................. 14 Table 9. Lab Control Spike Results.................................................................................................. 15 Table 10. Sample and QC Results..................................................................................................16 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 5 o f 122 Study Information Sponsor 3M Company Sponsor Representative Michael A. Santoro 3M Company 3M Building 0236-01-B-10 St. Paul, MN 55144 Study Director Jaisimha Kesari, P.E., DEE Weston Solutions, Inc. 1400 Weston Way West Chester, PA 19380 EOS-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Study Location Testing Facilities Exygen Research 3058 Research Drive State College, PA 16801 3M Environmental Laboratory 3M Building 2-3E-09 935 Bush Avenue St. Paul, MN 55144 Study Dates Study Initiation: 11/5/2004 Interim Experimental Initiation: January 11, 2006 Interim Experimental Completion: January 27, 2006 Interim Study Completion: Date of interim report signing Location of Archives All original raw data, protocol, and analytical report have been archived at the 3M Environmental Laboratory according to 40 CFR Part 792. The test substance and analytical reference standard reserve samples are archived at the 3M Environmental Laboratory according to 40 CFR Part 792. E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 7 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 S ummary and Introduction The 3M Environmental Laboratory extracted and analyzed water samples collected from six Northern Alabama potable water systems by Weston Solutions personnel on December 13 and 14, 2005. Samples were submitted for analysis as part of 3M Environmental Laboratory project number E05-0210 (3M Decatur Fluorochemical GLP Monitoring Program, Interim Report #18: Analysis of Water Samples from Northern Alabama Potable Water Systems, December 2005). Water samples were analyzed for perfluorobutane sulfonate (PFBS), perfluorohexane sulfonate (PFHS), and perfluorooctane sulfonate (PFOS) using 3M Environmental Laboratory Method ETS 8-154.1 "Determination of Perfluorinated Acids, Alcohols, Amides, and Sulfonates in Water by Solid Phase Extractions and High Performance Liquid Chromatography/Mass Spectrometry" in accordance with Exygen Research Protocol P0001131 (Attachment C). ETS 8-154.1 falls under the 3M Environmental Laboratory's current ISO/IEC 17025-1999 scope of accreditation and the results reported herein were compliant with this accreditation. The analytical start date for this interim report was January 11, 2006 and the analytical termination date was January 27, 2006. Sample containers were prepared at the 3M Environmental Laboratory. Sample containers for each sampling location included a field sample, field sample duplicate, low field spike (0.1 ng/mL) and high field spike (1.0 ng/mL). Each empty container was marked with a "fill to here" line to produce a final sample volume of 450 mL. Containers designated for field matrix samples were fortified with an appropriate matrix spike solution containing the three target analytes prior to being sent to the field for sample collection. For the samples and field matrix spikes collected and analyzed for this interim report, PFOA [1,2 13C] was not included as a surrogate. This is a deviation from Protocol P0001130 Amendment #3. A record of deviation has been issued as is included in Attachment C. Table 1 below summarizes the sample results. The average between the sample and the sample duplicate is provided along with the relative percent difference (%RPD), if applicable. The limit of quantitation (LOQ) of PFBS, PFHS, and PFOS for the sample set was 0.0248 ng/mL, 0.0252 ng/mL, and 0.0249 ng/mL, respectively. All results for quality control samples prepared and analyzed with the samples will be reported and discussed elsewhere in this report. E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 8 o f 122 E05-0210 Interim Report *18 Northern Alabama Potable Water Systems, December 2005 Table 1. Sample Results Summary 3MLIMSID Sample Description E05-0210-92250 DPWS PW FW02 0 051213 E05-0210-92251 DPWS PW FW 02 DB 051213 Average Sample/Sample Duplicate %RPD E05-0210-92254 W MELPW FW 02 0 051213 E05-0210-92255 W M ELPW FW 02DB 051213 Average Sample/Sample Duplicate %RPD E05-0210-92262 MSTP PW FW 02 0 051214 E05-0210-92263 MSTP PW FW02 DB 051214 Average Sample/Sample Duplicate %RPD E05-0210-92266 FW TPPW FW 02 0 051214 E05-0210-92267 FWTP PW FW 02 DB 051214 Average Sample/Sample Duplicate %RPD E05-0210-92270 TVATP PW FW 02 0 051214 E05-0210-92271 TVATP PW FW02 DB 051214 Average Sample/Sample Duplicate %RPD E05-0210-92258 E05-0210-92259 SWTP PW FW02 0 051214 SWTP PW FW 02 DB 051214 Average Sample/Sample Duplicate %RPD ,1>PFBS Concentration (ng/mL) <0.0248 <0.0248 0.0248 <4,n a 0.0350 0.0321 0.0336 8.6 <0.0248 <0.0248 <0.0248 <4>n a <0.0248 <0.0248 <0.0248 <4>n a 0.0344 0.0347 0.0346 0.87 <0.0248 <0.0248 <0.0248 <4>n a mPFHS Concentration (ng/mL) <0.0252 <0.0252 <0.0252 <4>n a <0.0252 <0.0252 <0.0252 ,4,n a <0.0252 <0.0252 <0.0252 ,4>n a <0.0252 <0.0252 <0.0252 ,4>n a <0.0252 <0.0252 <0.0252 <4>n a <0.0252 <0.0252 <0.0252 <4,n a mPFOS Concentration (ng/mL) <0.0249 <0.0249 <0.0249 <4,n a 0.0367 0.0367 0.0367 0.0 <0.0249 <0.0249 <0.0249 <4>n a <0.0249 <0.0249 <0.0249 <4>n a <0.0249 <0.0249 <0.0249 ,4>n a <0.0249 <0.0249 <0.0249 ,4>n a (1) The analytical uncertainty of the PFBS results Is 1005.3% based on method accuracy and precision. All results and averages are presented with three significant figures, %RPD to two significant figures. Sample concentrations, averages, and %RPD values may vary slightly from the raw data. Samples with reported results less than the LOQ of 0.0248 ng/mL may or may not have PFBS present. Results less than the LOQ are not quantified. (2) The analytical uncertainty of the PFHS results is 1009.3% based on method accuracy and precision. All results and averages are presented with three significant figures, %RPD to two significant figures. Samples with reported results less than the LOQ of 0.0252 ng/mL may or may not have PFHS present. Results less than the LOQ are not quantified. (3) The analytical uncertainty of the PFOS results is 10011% based on method accuracy and precision. All results and averages are presented with three significant figures, %RPD to two significant figures. Sample concentrations, averages, and %RPD values may vary slightly from the raw data. Samples with reported results less than the LOQ of 0.0249 ng/mL may or may not have PFOS present. Results less than the LOQ are not quantified. (4) NA: Not applicable. %RPD values not determined when concentrations for the sample and/or sample duplicate are below the stated LOQ. E05-0210 Interim Report *18 Northern Alabama Potable Water Systems, December 2005 Page 9 o f 122 EOS-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Test Samples Water samples from six locations were received at the 3M Environmental Laboratory on December 16, 2005 from Tim Frinak of Weston Solutions, Inc. Sample containers were received in satisfactory condition at ambient temperatures. A total of twenty-seven containers were submitted for analysis. The samples were logged in by 3M Environmental Laboratory professional services personnel and placed in refrigerated storage on December 16, 2005 until they were removed for extraction on January 11, 2006. Table 2 below provides the codes for the given sample descriptions. Table 2. Sample Description Codes.________________________________________ Sample Description Abbreviation Definition DPWS WMEL SWTP MSTP FWTP TVATP Decatur Public Water System West Morgan/East Lawrence Water Treatment Plant, Lawrence County, AL Sheffield Water Treatment Plant, Sheffield AL Muscle Shoals Treatment Plant, Muscle Shoals, AL Florence Water Treatment Plant, Florence, AL Tennessee Valiev Authority Treatment Plant, Muscle Shoals, AL PW Potable Water FW Finished Water 0 Matrix Sample DB Duplicate LS Low Spike HS High Spike Reference Substances Table 3 lists the pertinent information regarding the reference substance used for this study. _____ Table 3. Study Reference Substances.__________________ ___________ ________ Reference Substance PFBS PFHS PFOS Chemical Name Perfluorobutanesulfonate Perfluorohexanesulfonate Perfluorooctanesulfonate Chemical Formula Identifier <1,Source Expiration Date Storage Conditions Chemical Lot Number TCR Number C4 F9SO3 K' CAS # 29420-49-3 3M 1/17/2007 Frozen 2 TCR-281 (99131-039) C6 F,3S 0 3'K* CAS # 3871-99-6 3M 10/18/2006 Frozen NB# 120067-69 TCR-83 (SE036) CsF^SOs-K* CAS #2795-39-3 3M 8/31/2006 Frozen 171 TCR-696 Physical Description White Powder White Powder White powder Purity 97.3% 98.6% 86.4% ^'Solubility 54,400 ppm No Information available (1) Documentation regarding synthesis of the test substances is located at the source. 680 ppm (2) All test substances are believed to be soluble in water at the levels to be Investigated. No visual precipitates observed. E0S-0210 Interim Report #18 Northern Alabama Potable Water Systems, December2005 Page 10 o f 122 E0S-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Method Summaries Sample Collection Samples were collected on December 13-14, 2005 in rinsed NalgeneTM (low-density polyethylene) bottles prepared at the 3M Environmental Laboratory on December 9, 2005. Containers designated for field matrix spike samples were spiked in the laboratory with a known volume of an appropriate matrix spiking solution containing the target analytes. Four collection bottles were associated with each individual sampling location: sample, sample duplicate, low level field matrix spike (LS), and high level field matrix spike (HS). Additionally, one "Trip Blank" sample set was prepared. A Trip Blank set consisted of three individual bottles: trip blank sample, trip blank low spike, and trip blank high spike. The matrix spike levels for the trip blanks were the same as the samples. Table 4 below details the spike amounts for the four bottles comprising the sample location set. Table 4. Sample Collection Information mBottle Description Nominal Final Volume Collected (mL) Sample Sample Dup Low Field Matrix Spike (LS) High Field Matrix Spike (HS) 450 450 450 450 Final Concentration (ng/mL) PFBS PFHS PFOS NA NA 0.0992 0.992 NA NA 0 .1 0 1 1 .0 1 NA NA 0.0997 0.997 (1) The sample description was assigned in the field by Weston Solutions personnel. Preparatory and Analytical Methods Extraction All samples, calibration standards, and associated quality control samples were extracted using the procedure outlined in ETS-8-154.1 "Determination of Perfluorinated Acids, Alcohols, Amides, and Sulfonates in Water by Solid Phase Extractions and High Performance Liquid Chromatography/Mass Spectrometry". Briefly, 40 mL of sample was loaded onto a pre-conditioned Waters C18 solid phase extraction (SPE) cartridge (Sep-Pak, 6 cc) using a vacuum manifold. The loaded cartridges were then eluted with 5 mL of methanol. This extraction procedure concentrates the samples by a factor of eight. (Initial volume = 40 mL, final volume = 5 mL). Samples were initially extracted on January 11, 2006 and analyzed on January 14, 2006. Because associated laboratory quality control samples did not meet method criteria, samples were reextracted along with a new calibration curve and all associated quality control samples on January 23, 2006. The second set of extracts were analyzed on January 26, 2006. All results reported here are from the second extraction set. Analysis All sample and quality control extracts were analyzed for PFBS, PFHS, and PFOS using high performance liquid chromatography/ tandem mass spectrometry (HPLC/MS/MS). Pertinent instrument parameters, the liquid chromatography gradient program, and the specific mass transitions analyzed are described in the tables below. E0S-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 11 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Table 5. Instrument Parameters Instrument Name Liquid Chromatograph Guard column Analytical column Injection Volume Mass Spectrometer Electrode Ion Source Polarity Software ETSOIlie Agilent 1100 Two Betasil C18 (2.1 mm X 100 mm), 5 pm linked in series before the injection port Betasil C18 <2.1 mm X 100 mm), 5 pm 5pL Applied Biosystems API 4000 Q Trap Z-spray Turbo Spray Negative Analyst 1.4.1 Table 6. Liquid Chromatography Gradient Program Step Number Total Time (min) 00 1 1 .0 2 14.5 3 15.5 4 16.5 5 2 0 .0 Flow Rate (fiL/min) 300 300 300 300 300 300 Percent A (2 mM ammonium acetate) 80.0 80.0 1 0 .0 1 0 .0 80.0 80.0 Percent B (Methanol) 2 0 .0 2 0 .0 90.0 90.0 2 0 .0 2 0 .0 Table 7. Mass Transitions Analyte (1)PFBS (1)PFHS <1)PFOS Mass Transition Q1/Q3 299/80 299/99 399/130 399/99 399/80 499/130 499/99 499/80 Dwell Time (msec) 150 150 150 150 150 150 150 150 (1) The individual transitions were summed to produce a `total ion chromatogram' (TIC) for the given analyte. The resultant TIC was used for quantitation. E0S-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 12 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 A nalytical Results Calibration Calibration standards were prepared by spiking known amounts of stock solutions containing the target analytes into 40 mL of ASTM type I water. Each spiked water standard was then extracted in the same manner as the collected samples. A total of twelve spiked standards ranging from 0.025 ng/mL to 25 ng/mL (nominal) were prepared. A quadratic, 1/x weighted, calibration curve was used to fit the data. The data was not forced through zero during the fitting process. Calculating the standard concentration using the peak area counts and the resultant calibration curve confirmed accuracy of each curve point. Each extracted calibration standard used to generate the final calibration curve met the method calibration accuracy requirement of 10025% (10030% for the LOQ standard). The correlation coefficient was greater than 0.999 for all analytes. Limit of Quantitation (LOQ) The LOQ for this analysis, as defined in ETS-8-154.1, is the lowest non-zero calibration standard in the curve in which the area counts are at least twice those of the method blank(s). This will be presented and discussed in more detail in the section below regarding method blanks. The LOQ for PFBS, PFHS, and PFOS was 0.0248 ng/mL, 0.0252 ng/mL, and 0.0249 ng/mL, respectively. Blanks Three types of blanks were prepared and analyzed with the samples: method blanks, solvent blanks, and field/trip blanks. Each blank type is described below. Solvent Blank Several methanol solvent blanks were analyzed to assess system contamination and/or instrument carryover. One methanol solvent blank analyzed immediately after the highest calibration standard produced PFOS area counts greater than half the LOQ standard. However, the solvent blank analyzed immediately after the high blank demonstrated that the likely instrument carryover observed in the preceding injection had diminished. Analyte peak area counts in all other solvent blank samples were less than half the area counts of the calibration standard used to establish the LOQ for all analytes. Method Blank Eight method blanks were prepared by loading 40 mL of ASTM Type I water onto a C18 SPE cartridge and eluting with 5 mL of methanol using the same extraction procedure as the samples. Method blanks were prepared to evaluate the levels of background contamination in the overall extraction process (glassware, SPE cartridges, etc.) Table 8 lists the PFOA area counts for the method blanks and the corresponding LOQ standard. EOS-0210 Interim Report *18 Northern Alabama Potable Water Systems, December 2005 Page 13 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Table 8. Method Blank Area Counts. Sample ID Sample Comment PFBS PFHS Area Counts PFOS MB-060123-1 Method Blank-1 7923 8472 MB-060123-2 Method Blank-2 18778 25105 M B -0 6 0 123-3 Method Blank-3 8061 8900 MB-060123-4 Method Blank-4 6404 11382 MB-060123-5 Method Blank-5 5688 7332 MB-060123-6 Method Blank- 6 14035 13232 M B -0 6 0 123-7 Method Blank-7 5693 4863 MB-060123-8 Method Blank- 8 4299 7087 Area counts of the highest method blank multiplied by two 37556 50210 Area Counts of the LOQ standard 179926 162287 (1) PFOS area countsforMethod Blank-5 determined to be a statistical outlier using Dixon's Q-tesl 21276 25856 16771 19740 (1,55137 21206 15055 18065 |2,51712 102452 (2) Highest method blank area count for PFOS is 25856 (Method Blank-2) when Method Blank-5 is excluded as on outlier. Trip Blank Prior to sample collection, one sample container was filled with 450 mL of ASTM Type I water, sealed, and shipped to the sample collection site along with the empty containers. This sample was analyzed as the field/trip blank. The trip blank serves as an additional method blank that account for any storage conditions and/or holding time issues that the samples may experience. The resultant field blank concentrations for PFBS, PFHS, and PFOS were <0.0248 ng/mL, <0.0252 ng/mL, and <0.0249 ng/mL, respectively. System Suitability Five replicate injections of the 1.0 ppb (nominal concentration) extracted calibration standard were analyzed at the beginning and end of the analytical sequence to demonstrate overall system suitability. The relative standard deviation (RSD) of the peak area counts and the RSD of the peak retention times were less than 5% and 2%, respectively, for all analytes which met method criteria. Continuing Calibration During the course of the analytical sequence, several continuing calibration verification samples (CCVs) were analyzed to confirm that the instrument response and the initial calibration curve were still in control. ETS-8-154.1 states that all CCV recoveries must be within 25% for the data to be acceptable. All analyte CCV recoveries met method criteria of 25%. Lab Control Spikes (LCSs) Replicate low (0.05 ng/mL nominal concentration) and high (0.5 ng/mL nominal concentration) lab control spikes were prepared and extracted on the same day as the samples. LCSs were prepared by spiking known amounts of the analytes into 40 mL of ASTM Type I water to produce the desired concentration. The spiked water samples were extracted and analyzed in the same manner as the samples. Individual LCS results, along with the average and percent RSD, for the extraction set are presented in the data table below. All LCS recoveries met method criteria for both accuracy and precision. E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 14 o f 122 EOS-0210 Interim Report 018 Northern Alabama Potable Water Systems, December 2005 Table 9. Lab Control Spike Results. mPFBS WPFHS (1>PFOS Lab ID Spiked Cone. (ng/mL) Calc. Cone. (ng/mL) % Spiked Recovery Cone. (ng/mL) Calc. Cone. (ng/mL) % Spiked Cone. Recovery (ng/mL) Calc. Cone. (ng/mL) % Recovery LCS-060123-1 0.0496 0.0469 94.6 0.0503 0.0459 91.2 0.0499 0.0453 LCS-060123-2 0.0496 0.0473 95.4 0.0503 0.0456 90.8 0.0499 0.0469 LCS-060123-3 0.0496 0.0483 97.4 0.0503 0.0465 92.4 0.0499 0.0429 LCS-060123-4 0.496 0.485 97.8 0.503 0.500 99.4 0.499 0.477 LCS-060123-5 0.496 0.473 95.4 0.503 0.472 93.8 0.499 0.462 LCS-060123-6 0.496 0.497 100 0.503 0.488 97.0 0.499 0.475 Average `/Recovery 96.8% 1 2.2% 94.1% 3.6% 92.3%3.9% %RSD (1) Table displays rounded values for all concentration and percent recovery values (3 significant figures) and %RSD (2 significant figures) using EPA rounding rules. Reported values may vary slightly from the raw data. 90.8 94.0 8 6 .0 95.6 92.6 95.2 Sample Duplicates Because a field sample duplicate (separate container) was collected at each sampling location, duplicate/replicate extractions of a given sample were not performed. Overall method precision was determined using LCSs. Field Matrix Spikes Low (nominal concentration of 0.1 ng/mL) and high (nominal concentration 1.0 ng/mL) field matrix spikes were collected at each sampling point to verify that the analytical method is applicable to the collected matrix. Field matrix spike recoveries within 10030% confirm that "unknown" components in the sample matrix do not interfere with the extraction and analysis of the analytes of interest. Field matrix spikes will be presented in the next section with the sample data. Data Summary and Discussion Table 10 below summarizes the sample results and field matrix spike recoveries for all of the sampling locations plus the trip blank. All field matrix spikes produced recoveries within 100+30% demonstrating that the analytical method was appropriate for the matrices analyzed. E05-0210 Interim Report 018 Northern Alabama Potable Water Systems, December 2005 Page 15 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Table 10. Sample and QC Results. Location #1 DPWSPWFW01 mPFBS mPFHS mPFOS 3M LIMS ID Description E05-0210-92250 DPWS PW FW02 0 051213 E05-0210-92251 DPWS PW FW02 DB 051213 E05-0210-92252 DPWS PW FW02 LS 051213 E05-0210-92253 DPWS PW FW2 HS 051213 Average Sample Concentration (ng/mL) %RPtf4> Cone. (ng/mL) %Recovery <0.0248 <0.0248 0.0949 0.978 <0.0248 NA NA 95.6 98.6 Cone. (ng/mL) %Recovery <0.0252 <0.0252 0.0971 NA NA 96.5 1 .0 2 <0.0252 101 Cone. (ng/mL) %Recovery <0.0249 <0.0249 0.0853 0.918 <0.0249 NA NA 85.5 92.0 Location #2 WMEL PWFW01 IVPFBS 3M LIMS ID Description E05-0210-92254 WMEL PW FWQ2 0 051213 E05-0210-92255 W M ELPW FW 02D B 051213 E05-0210-92256 WMEL PW FW02 LS 051213 E05-0210-92257 W M ELPW FW 02 HS 051213 Average Sample Concentration (ng/mL) %RPCf4> Cone. (ng/mL) %Recovery 0.0350 NA 0.0321 NA 0.130 97.2 1 .0 2 99.4 0.0336 i 8.6% mPFHS mPFOS Cone. (ng/mL) %Recovery <0.0252 <0.0252 0.0990 0.950 <0.0252 NA NA 98.8 94.4 Cone. (ng/mL) %Recovery 0.0367 NA 0.0367 NA 0 .1 2 1 84.5 0.883 84.8 0.0367 0.0% Location #3 MSTPPWFW01 ,1>PFBS 3M UMS ID Description E05-0210-92262 MSTP PW FW02 0 051214 E05-0210-92263 MSTP PW FW02 DB 051214 E05-0210-92264 MSTP PW FW02 LS 051214 E05-0210-92265 MSTP PW FW02 HS 051214 Average Sample Concentration (ng/mL) %RP4> Cone. (ng/mL) %Recovery <0.0248 <0.0248 0.115 0.934 <0.0248 NA NA 116 94.1 mPFHS Cone. (ng/mL) ^Recovery <0.0252 <0.0252 0.0930 0.958 <0.0252 NA NA 92.3 95.2 mPFOS Cone. (ng/mL) %Recovery <0.0249 <0.0249 0.0922 0.826 <0.0249 NA NA 92.4 82.8 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 18 of 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Table 10 Continued. Location #4 FWTP PW FW01 <1>PFBS 3M LIMS ID Description E05-0210-92266 FWTP PW FW02 0 051214 E05-0210-92267 FWTP PW FW02 DB 051214 E05-0210-92268 FWTP PW FW02 LS 051214 E05-0210-92269 FWTP PW FW02 HS 051214 Average Sample Concentration (ng/mL) %RPDft> Conc. (ng/mL) '/Recovery <0.0248 <0.0248 0.118 0.962 <0.0248 NA NA 119 96.9 WpFHS Conc. (ng/mL) %Recovery <0.0252 <0.0252 0.0900 0.927 < 0.0252 NA NA 89.3 92.1 (3Ipfo s Conc. (ng/mL) %Recovery <0.0249 <0.0249 NA NA 0 .1 0.811 <0.0249 100 81.3 Location #5 TVATP PW FW01 (VPFBS 3M LIMS ID Description E05-0210-92270 TVATP PW FW02 0 051214 E05-0210-92271 TVATP PW FW02 DB 051214 E05-0210-92272 TVATP PW FW2 LS 051214 E05-0210-92273 TVATP PW FW02 HS 051214 Average Sample Concentration (ng/mL) %RPtl> Conc. (ng/mL) %Recovery 0.0344 NA 0.0347 NA 0.132 98.2 0.910 8 8 .2 0.03460.87% mPFHS mPFOS Conc. (ng/mL) %Recovery <0.0252 <0.0252 0.0909 0.900 <0.0252 NA NA 90.4 89.5 Conc. (ng/mL) '/.Recovery <0.0249 <0.0249 0.104 0 .8 6 8 <0.0249 NA NA 104 87.0 WpFBS mPFHS &PFOS 3M UMS ID Description E05-0210-92258 SW TPPWFW 02 0 051214 E05-0210-92259 SWTP PW FW02 DB 051214 E05-0210-92260 SWTP PW FW02 LS 051214 E05-0210-92261 SWTP PW FW02 HS 051214 Average Sample Concentration (ng/mL) %RPDf4> Conc. (ng/mL) %Recovery <0.0248 <0.0248 NA NA 0 .1 1 0 0.91 <0.0248 111 91.5 Conc. (ng/mL) %Recovery <0.0252 <0.0252 0.0836 0.927 <0.0252 NA NA 83.1 92.1 Conc. (ng/mL) %Recovery <0.0249 <0.0249 0.0917 0.837 <0.0249 NA NA 91.9 83.9 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 17 of 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Table 10 Continued. Trip Blank <1>PFBS mPFHS VPFOS 3MUIHSID E05-0210-92247 E05-0210-92248 E05-0210-92249 Description DPW SPW TRIP 0 051213 DPWS PW TRIP LS 051213 DPWS PW TRIP HS 051213 Cone. (ng/mL) < 0.0248 0.0921 0.946 %Recovery NA 92.8 95.3 Cone. (ng/mL) <0.0252 0.0921 0.977 %Recovery NA 91.6 97.1 Cone. (ng/mL) < 0.0249 0.0837 0.898 %Recovery NA 83.9 90.0 (1) The analytical uncertainty of the PFBS resuts is 1005.3% based on method accuracy and precision. All results and averages are presented with three significant figures. %RPD to two significant figures. Sample concentrations, averages, and %RPD values may vary slightly from the raw data. Samples with reported results less than the LOQ of 0.0248 ng/mL may or may not have PFBS present. Results less than the LOQ are not quantified. (2) The analytical uncertainty of the PFHS resuts is 1009.3% based on method accuracy and precision. All results and averages are presented with three significant figures, %RPD to two significant figures. Sample concentrations, averages, and %RPD values may vary slightly from the raw data. Samples with reported results less than the LOQ of 0.0252 ng/mL may or may not have PFHS present. Results less than the LOQ are not quantified.. (3) The analytical uncertainty of the PFOS resuts is 100111 % based on method accuracy and precision. All results and averages are presented with three significant figures, %RPO to two significant figures. Sample concentrations, averages, and %RPD values may vary slightly from the raw data. Samples with reported results less than the LOQ of 0.0249 ng/mL may or may not have PFOS present. Results less than the LOQ are not quantified. (4) NA: Not applicable. %RPD values not determined when concentrations for the sample and sample duplicate are below the stated LOQ. E0S-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 18 o f 122 EOS-0210 Interim Report #18 Northern Alabama Potable Water Systems, December2005 Statistical Methods and Calculations Statistical methods used to interpret sample results include averages and standard deviations. The Analyst software program calculated sample concentrations using resultant analyte peak areas and the established quadratic, 1/x weighted, calibration curve. None of the samples analyzed for this interim report required dilution. Sample calculations and equations used to report method accuracy and precision are described below. Accuracy and Precision Equations Calculated Concentration LCS/Surrogate Percent Recovery = 100% Spike Concentration (Spiked Sample Concentration - Average Concentration : Field Sample & Field Sample Pup.) , 1 Sample Spike Recovery = Spike Concentraton % RSD , (Relative Standjard ... . Deviation) standard deviation of replicates ------------------------------------ ---------- , ..... 100% replicate average . Absolute difference between sample duplicates % RPD (Relative Percent Difference) --------------------------------- --------------- Z--.-- --------- * 100% average sample concentration Determination of Analytical Uncertainty Both the accuracy (percent recovery) and precision (%RSD) of the lab control spikes are used to estimate the analytical uncertainty for a given analyte. The measured precision (%RSD) is then used to determine the range of the accuracy. The following example demonstrates how the analytical uncertainty, as defined here, is calculated for PFOS. Example: First, the accuracy range is determined by multiplying the average recovery by the precision. 92.35 (average accuracy)*0.0389 (precision) = 3.59 (accuracy range) Next, the accuracy range is added and subtracted from the average accuracy. 92.35 + 3.59 = 95.94; 92.35 -3.59 = 88.76 Thus, LCS accuracy results range from 88.76% to 95.94%. The absolute difference of the low and high ends of this range, when compared 100%, are then calculated. 100%-95.94 = 4.06% 100%-88.76%= 11.23%. The most conservative (largest) absolute difference is then used as the analytical uncertainty for the given analyte. Therefore, the analytical uncertainty for PFOS as defined here, is given as 10011% (2 significant figures) for the data associated with this project. E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 19 o f 122 E0S-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Statement of Conclusion Sample results for this project were presented in Table 1. Laboratory control spikes were used to determine the method accuracy and precision for the target analytes for this data set. The accuracy and precision were then used to estimate the analytical uncertainty for each of the analytes (PFBS: 1005.3%; PFHS: 1009.3%, and PFOS: 10011%). Recoveries of field matrix spiked samples demonstrated that the overall analytical method was appropriate for the matrix collected. References Rorabacher, David B. "Statistical Treatment for Rejection of Deviant Values: Critical Values of Dixon's `Q' Parameter and Related Subrange Ratios at the 95% Confidence Level." Anal. Chem. 1991, 6 3 ,139-146. List of Attachments Attachment A: Selected Chromatograms and Calibration Curves Attachment B: Extraction and Analytical Methods Attachment C: Protocol and Protocol Amendments E05-0210 Interim Report *18 Northern Alabama Potable Water Systems, December 2005 Page 20 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Signature Page We certify that this report is a true and complete representation of the data for this study: William K. Reagen, Ph.D., AAA<Mm) Michael A. Santoro O 3M Environmental Laboratory Manager Date Sponsor Representative Date 05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 21 o f 122 E0S-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Attachment A: Sample Chromatograms and Calibration Curves E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 22 o f 122 EOS-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 23 o f 122 E05-0210 Interim Report # f 8 Northern Alabama Potable Water Systems, December 2005 Page 24 o f 122 olile E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 25 o f 122 EOS-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 26 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 27 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 28 o f 122 EOS-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 29 o f 122 EOS-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 30 o f 122 EOS-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 E05-0210 Interim Report #18 Northern Alabama Potable Wafer Systems, December 2005 Page 31 o f 122 EOS-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 E0S-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 32 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 33 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Attachment B: Extraction and A nalytical Methods E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 34 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December2005 3M Environm ental Laboratory Method Determination o f Perfluorinated Acids, Alcohols, Amides, and Sulfonates In Water By Solid Phase Extraction and High Performance Liquid Chromatography/Mass Spectrometry Method Number: ETS-8-154.1 Adoption Date: 28 Apr 2000 Revision Date: 5 May, 2003 Effective Date: 5 May, 2003 Approved By: W illiam K. Reagen Manager O S '/c S ~ /o 3 Date ETS-8-154.1 Page 1 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 35 o f 122 E0S-0210 Interim Report #18 Northern Alabama Potable Water Systems, December2005 1 Scope and Application This method was validated for the collection, extraction, and analytical procedure for the determination of Perfluorooctane sulfonate (PFOS), Perfluorooctane sulfonylamide (FOSA), and Perfluorooctanoate (PFOA) in groundwater, surface water, and drinking water samples. This method may also be applied to the determination of other perfluorinated acids, alcohols, amides, and sulfonates in similar matrices, as long as the defined QC elements are satisfied and with the understanding that the method is not validated for compounds outside the scope of the original protocol This method is based in part on the report "Method ofAnalysis for the [Determination of Perfluorooctane sulfonate (PFOS), Perfluorooctane sulfonylamide (PFOSA), and Perfluorooctanoate (POAA) inWater" (see Section 17), as developed and validated by Exygen Research (formerly Centre Analytical Laboratories, Inc.). 2 Method Summary Water samples are collected from a site of interest and shipped cold to an analytical facility. Perfluorinated acids, alcohols, amides, and sulfonates are extracted from 40mL water samples using C18 solid phase extraction (SPE) cartridges. The compounds are eluted from the C18 cartridge, using methanol. Separation, identification, and measurement are accomplished by high performance liquid chromatography/ tandem mass spectrometry (HPLC/MS/MS) analysis. High performance liquid chromatography/mass spectrometry (HPLC/MS) may be used if the defined QC elements are satisfied. The concentration of each identified component is measured by comparing the MS response of the quantitation ion produced by that compound to the MS response of the quantitation bn produced by the same compound in an extracted calibration standard (external standard). 3 Definitions 3.1 A nalytical Sam ple A portion of an extracted Laboratory Sample prepared for analysis. 3.2 Calibration Standard A solution prepared from the Working Standard (W S) and extracted according to this method. The calibration standard solutions are used to calibrate the instrument response with respect to analyte concentration. 3.3 Duplicate Sample (DS) A DS is a separate aliquot of a sample, taken in the analytical laboratory that is extracted and analyzed separately with identical procedures. Analysis of DSs compared to that of the first aliquot give a measure of the precisbn associated with laboratory procedures, but not with sample collection, preservation, or storage procedures. 3.4 Field Blank Control Sample (FB) ASTM Type I water placed in a sample container in the laboratory and treated as a sample in all respects, including exposure to sampling site conditions, storage, preservation and all analytical procedures. The purpose of the FB is to determine if test substances or other interferences are present in the field environment ETS-8-154.1 Page 2 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 36 o f 122 E0S-0210 Interim Report #18 Northern Alabama Potable Water Systems, December2005 3.5 Field Duplicate (FD) A sample collected in duplicate at the same time as the sample and placed under identical circumstances and treated exactly the same throughout field and laboratory procedures. Analysis of FD compared to that of the first sample gives a measure of the precision associated with sample collection, preservation and storage, as well as with laboratory procedures. 3.6 Field Matrix Spike (FMS) A sample collected in duplicate to which known quantities of the target analytes are added inthe field at the time of sample collection. Alternatively, the known quantity of target analytes may be added to the sample bottle in the laboratory before the bottles are sent to the field. A known, specific volume of sample must be added to sample container without rinsing. This may be accomplished by making a "fill to this level' line on the outside of the sample container. The FMS should be spiked at approximately 50 150% of the expected analyte concentration in the sample. If the expected range of analyte concentrations is unknown, a low and a high spike may be prepared to increase the likelihood that a spike at an appropriate range is made. The FMS is analyzed to ascertain if any matrix effects, interferences, or stability issues may complicate the interpretation of the sample analysis. 3.7 Field Spike Control Sam ple (FSCS) An aliquot ofASTM Type I water to which known quantities of the target analytes are added in the field at the time of sample collection (at an appropriate concentration to be determined by the project lead) or in the laboratory priorto the shipment of the collection bottles. The FSCS is extracted and analyzed exactly like a sample to determine whether a loss of analyte could be attributed to sample storage and/or shipment. A low and high FSCS may be appropriate when expected sample concentrations are not known. 3.8 Laboratory Control Sample (LCS) An aliquot ofASTM Type I water to which known quantities ofthe target analytes are added inthe laboratory. Two levels are included, one at the LLOQ (approx. 25 pg/mL), the other at a concentration of approx. 100 250 pg/mL or another concentration to be determined by the project lead. The LCS is extracted and analyzed exactly like a laboratory sample to determine whether the methodology is in control, and whether the laboratory is capable of making accurate measurements at the required method detection limit and higher. 3.9 Laboratory Sample A portion of a sample received from the field for testing. 3.10 Limit of Detection (LOD) The LOD is the lowest concentration of an analyte that can be measured and reported with 99% confidence that the analyte concentration is greater than zero. If required, the LOD may be determined in several ways, including signal-to-noise ratio and statistical calculations. 3.11 Limit o f Quantitation (LOQ) The LOQ for a dataset is the lowest concentration (LLOQ) or highest concentration (ULOQ) that can be reliably achieved within the specified limits of precision and accuracy during routine operating conditions. Note: For many analytes, the LLOQ analyte concentration is selected as the lowest non-zero standard in the calibration curve to simplify data reporting. Sample LLOQs are matrix-dependent. 3.12 Matrix Spike (MS) ETS-8-154.1 Page 3 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. EOS-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 37 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 A matrix spike is an aliquot of a sample, to which known quantities of target analytes are added in the laboratory. The MS is extracted and analyzed exactly like a laboratory sample to determine whether the sample matrix contributes bias to the analytical results. The background concentrations of the analytes in the sample matrix must be determined in a separate aliquot and the measured values inthe MS corrected for background concentrations. 3.13 Method Blank An aliquot ofASTM Type I water that is treated exactly like a laboratory sample including exposure to all glassware, equipment, solvents, and reagents that are used with other laboratory samples. The method blank is used to determine iftest substances or other interferences are present inthe laboratory environment, the reagents, or the apparatus. 3.14 Method Detection Lim it (MDL) Determination A MDL is the statistically calculated minimum amount of an analyte that can be measured with 99% confidence that the reported value is greater than zero. One of several processes that may be used to establish a LOD value is found in40 CFR Part 136 Appendix B. 3.15 Sample A sample is a small portion collected from a larger quantity o f m aterial intended to represent the original source material. 3.16 Spiking Stock Standard (SSS) A solution prepared from stock standards used to prepare the working standard. 3.17 Stock Standard (SS) A concentrated solution of a single analyte prepared in the laboratory with an assayed reference compound. 3.18 Working Standard (WS) A solution of several analytes prepared inthe laboratoryfrom SSs and diluted as needed to prepare calibration standards and other required analyte solutions. 4 Warnings and Cautions 4.1 Health and Safety The acute and chronictoxicity of the standards for this method have not been precisely determined; however, each should be treated as a potential health hazard. Unknown samples may contain high concentrations of volatile toxic compounds. Sample containers should be opened in a hood and handled with gloves to prevent exposure. The laboratory is responsible for maintaining a safe work environment and a current awareness of local regulations regarding the handling ofthe chemicals used in this method. A reference file of material safety data sheets (MSDS) should be available to all personnel involved in these analyses. 4.2 Cautions None 5 Interferences ETS-8-154.1 Page 4 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 38 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 During extraction and analysis, major potential contaminant sources are reagents and solid phase extraction devices. All materials used in the analyses shall be demonstrated to be free from interferences under conditions of analysis by running method blanks. Parts and supplies that contain Teflon should be avoided due to the possibility of interference and/or contamination. These may include, but are not limited to: wash bottles, Teflon lined caps, autovial caps, HPLC parts, etc. The use of disposable micropipettes or pipettes to aliquot standard solutions is recommended to make calibration standards and matrix spikes. 6 Instrumentation, Supplies, and Materials Note: Brand names, suppliers, and part numbers are for illustrative purposes only. Equivalent performance may be achieved using apparatus and materials other than those specified here, but demonstration of equivalent performance that meets the requirements of this method is the responsibility of the laboratory performing the analysis. 6.1 Instrum entation Balance, analytical (display at least 0.0001 g), Mettler HPLC/MS/MS or HPLC/MS system, as described in Section 10. 6.2 Supplies and Materials. Sample collection bottles--LDPE (e g., NalgeneTM) narrow-mouth bottles with screw cap. Note: Do not use Teflon bottles or Teflon lined caps. Coolers for sample shipment Ice for sample shipment. Vacuum pump, BOchi. Visiprep vacuum manifold, Supelco. Sep Pak Vac 6cc (1g) tC18 cartridges (part # WAT 036795),Waters. 50mL disposable polypropylene centrifuge tubes, VWR. 15mL disposable polypropylene centrifuge tubes, VWR. Disposable micropipettes (50 100pL, 100 200pL), Drummond. Class A pipettes and volumetric flasks, various. Hypercarb drop-in guard column (4mm) (part# 844017 400), Keystone. Stand-alone drop-in guard cartridge holder, Keystone. 125mL LDPE narrow-mouth bottles, Nalgene. 2mL clear HPLC vial kit (cat# 5181 3400), Agilent/Hewlett Packard. Standard lab equipment (graduated cylinders, disposable tubes, etc.), various. 7 Reagents and Standards______________________________________________ Note: Suppliers and catalog numbers are for illustrative purposes only. Equivalent performance may be achieved using chemicals obtained from other suppliers. Do not use a lessergrade of chemical than those listed. ETS-8-154.1 Page 5 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 39 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 7.1 Chem icals Methanol (MeOH), HPLC grade, JT Baker, Catalog No. JT9093 2. Am m onium Acetate, Reagent grade, Sigma-Aldrich, Catalog No. A-7330. ASTM Type I Water, prepared in-house. Sodium Thiosulfate, Reagent grade, JT Baker. 7.2 Standards Potassium perfluorooctane sulfonate Perfluorooctane sulfonylamide Ammonium perfluorooctanoate Others as required. 7.3 Reagent Preparation 250mg/mL sodium thiosulfate solution -- Dissolve 25g of sodium thiosulfate in 100mL reagent water. 40% methanol wash solution - Measure 400mL methanol and adjust volume to 1.0L with reagent water. 100mM ammonium acetate solution (Analysis)---Weigh 7.71g of ammonium acetate and dissolve in 1.0L of reagent water. Dilute the 100mM solution by a factor of 50 to make the 2mM ammonium acetate solution used formobile phase A. Note: Alternative volumes may be prepared as long as the ratios of the solvent to solute ratios are maintained. 7.4 Spiking Stock Standard (SSS) Preparation The following standard preparation procedure serves as an example and may be changed to suit the needs of a particular study. For example, pL volumes may be spiked into volumetric flasks when diluting stock solutions to appropriate levels. 100pg/mL each PFOS, PFOSA, and POAA SSSs--Weigh out 10mg of analytical standard (corrected for percent salt and purity--i.e., 10 mg CbF17S03K purity 90% = 8.35mg CaF^SOa") and dilute to 100mL with methanol in a lOOmL volumetric flask. Transfer to a 125mL LDPE bottle or other suitable container. Prepare a separate solution foreach analyte. Solutions may be stored in a refrigerator at 42C for a maximum period of 6 months from the date of preparation. 1pg/mL mixed SSS--Add 1.OmLeach ofthe 100pg/mLSSSs (from 7.4.1) to a 100mL volumetric flask and bring up to volume with methanol. 0.1pg/mL mixed SSS--Add 10.0mL ofthe 1.Opg/mL-mixed solution (from 7.4.2) to a 100mL volumetricflask and bring up to volume with methanol. 0.01pg/mL mixed SSS--Add 10.OmL ofthe 0.1pg/mL-mixed solution (from 7.4.3) to a 100mL volumetric flask and bring up to volume with methanol. Storage Conditions--Store all SSSs in a refrigerator at 402C for a maximum period of 6 months from the date of preparation. 7.5 Calibration Standards The following standard preparation procedure serves as an example and may be changed to suit the needs of a particular study, provided the concentrations are calculated correctly. ETS-8-154.1 Page 6 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 40 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 100|jg/mL each PFOS, PFOSA, and POAA stock standard solutions--Weigh out 10mg of analytical standard (corrected for percent salt and purity) and dilute to 100mL with methanol in a 100mL volumetric flask. Transfer to a 125mL LDPE bottle or other suitable container. Prepare a separate solution for each analyte. Store solutions in a refrigerator at 42C for a maximum period of 6 months from the date of preparation. 1pg/mL Working Standard--Add 1.0mL each of the 100pg/mL SS solutions (from 7.5.1) to a 100mL volumetric flask and bring up to volume with methanol. 0.1pg/mL Working Standard --Add 10.0mL of the 1.Opg/ml mixed solution (from 7.5.2) to a 100mL volumetric flask and bring up to volum e with methanol. 0.01|ig/mL Working Standard --Add 10.0mL ofthe 0.1 pg/mL mixed solution (from 7.5.3) to a 100mL volumetric flask and bring up to volume with methanol. Storage Conditions--Store all WSs in a refrigerator at 42C for a maximum period of 6 months from the date of preparation. Calibration Standard--Prepare calibration solutions inASTM Type I using the following table as a guideline: Final Calibration Concentration Volum e of Standard Volume, mL , of WS, pg/mL WS, pL of ASTM Type I W ater 0.0 0.010 0 100 40 40 0.010 0 .0 1 0 200 400 40 40 0.10 100 40 0.10 200 40 0.10 300 40 0.10 1.0 1.0 1.0 400 100 400 1000 40 40 40 40 Final Concentration of C alibration Standard, pg/mL, in ASTM Type I W ater 0 25 50 100 250 500 750 1000 2500 10000 25000 The standards are processed through the extraction procedure (Section 11), identical to the laboratory samples. The concentration of the calibration standard in the final extract is equal to 8X the initial concentration, due to the concentration of the standard during the extraction process. Storage Conditions--Store all extracted calibration standards in 15mL polypropylene tubes at 42C, for a maximum period of two weeks from the date of preparation 8 Sample Collection and Handling Note: Sampling equipment, including automatic samplers, must be free of Teflon tubing, gaskets, and other parts that may leach interfering analytes into the water sample. Automatic samplers that composite samples over time should use refrigerated polypropylene sample containers if possible. Sample bottles should not be rinsed before sample collection. Labeling: Each sample bottle must display information regarding the collection of that sample, the individual collecting the sample, and any matrix spike that has been added to the sample. ETS-8-154.1 Page 7 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 41 o f 122 E0S-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 This includes the volume and concentration of any spiking solution added and the volume and identification of any preservatives added inthe field. Spiking: The spring scheme will be clearly outlined in the sampling plan, including whether the samples will be spiked inthe field or in the laboratory prior to the shipment ofthe bottles to the site. If spiking is to be performed in the field, materials and specific instructions will be included in the sampling kit. Be sure to clearly label each bottle with spiking information if applicable. Tap Water: Open the tap and allow the system to flush until the water temperature (1510C) has stabilized (usually about two minutes). Adjust the flow to about 500mL/min and collect samples from the flowing stream. Ground Water. Purge the well of standing water using a pump or a bailer. Collect the sample directly from the pump or from the bailer. Surface W ater When sampling from an open body ofwater, fill the sample container with water from a representative area. Sample Dechlorination: All samples should be iced or refrigerated at 42C and kept inthe dark from the time of collection until extraction. Residual chlorine should be eliminated by adding 200pL of a 250mg/mL sodium thiosulfate solution to each tap-water sample and associated FB and FSCS (which may be placed in each bottle before leaving for the sampling site or done in the field.). Holding Time (HT): Results of the time/storage study of all target analytes showed that the three compounds are stable for 14 days in watersamples when the samples are dechlorinated and stored as described in the previous section (see also references in section 17). Therefore, laboratory samples must be extracted within 14 days and the extracts analyzed within 30 days of sample collection. If the HT exceeds 14 days, great care is used when evaluating field spikes to avoid misrepresentation of the sample concentration. 8.1 Field Blanks Process a Field Blank Control Sample (FB) along with each sample set (samples collected from the same general sample site at approximately the same time). At the laboratory, prior to sample collection, fill a sample containerwith ASTM Type I water, seal, and ship the FB to the sampling site along with the empty sample containers. Return the FB to the laboratory with the filled sample bottles. When sodium thiosulfate is added to samples, use the same procedure to presen the FB. 8.2 Field Duplicates Collect a Field Duplicate (FD) for every ten (10) samples collected or per each sampling set, if less than 10 samples are collected. Separate FDs must be collected for each type ofwater sample (ground, tap, etc.) collected. Collect the FD immediately after the sample. Preserve, store and ship FD using the same procedures as used for the samples. 8.3 Field Spike Control Sample (FSCS) A Field Spike Control Sample (FSCS) must be prepared for each sample shipment If multiple coolers are used to ship a set of samples, each cooler mustcontain a FSCS. At the laboratory, fill a sample container with 100mL ofASTM Type I water. Seal and ship to the sampling site along with the empty sample containers and FBs. Samples may either be spiked in the field or in the laboratory prior to shipment The method employed should be consistent throughout the study. If the samples are to be spiked in the field, be sure to send appropriate supplies and instructions for the field personnel to follow. ETS-8-154.1 Page 8 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December2005 Page 42 o f 122 EOS-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Seal and gently invert the FSCS to mix. Store and ship the FSCS using the same procedures as used for the samples Provide information on sample collection, preservation, shipment and storage. List applicable holding times. Include sample stability and extractstorage requirements. Reference the method used for sample preparation, if applicable. 8.4 Field Matrix Spike (FMS) A Field Matrix Spike (FMS) must be prepared for each sampling location. One unspiked sample from the same location must accompany the FMS to determine endogenous levels in the sample. The samples should be clearly identifiable as being from the same location. Samples may either be spiked in the field or in the laboratory prior to shipment. The method employed should be consistent throughout the study. If the samples are to be spiked in the field, be sure to send appropriate supplies and instructions for the field personnel to follow. 9 Quality Control and Data Quality Objectives Analytical results of the FB, FMS, FO, and FSCS should be evaluated at the conclusion ofthe study to help interpret the quality of sample data. Analytical results for these control/duplicate samples must be reported with the sample data. 9.1 Solvent Blanks Solvent blanks are analyzed with each sample set to determine contamination or carryover. Aliquots of methanol represent the solvent used for the standard curve and the sample extraction. Solvent blanks should have area counts that are less than 50% of the area count ofthe lowest calibration standard. Solvent blanks should be analyzed priorto and following each calibration curve, each set of system suitability samples, and after no more than 10 unknown sample extracts. If instrument carryover is a problem consecutive solvent blanks may be necessary. In this case the area counts of the solvent blanks should return to <50% of the lowest calibration standard prior to the injection of further standards or samples. 9.2 Method Blanks A method blank consists of an aliquot ofASTM Type I water, equal in volume to the samples, and extracted inthe same manner as the samples. At least two method blanks should be prepared and analyzed each day that extractions are performed for a particular study or project. When analyzed the area counts of these samples must be less than 50% of the area count of the lowest calibration standard. 9.3 Sample Replicates All samples, including field spikes, trip blanks, etc., should be extracted at least in duplicate, and in triplicate ifdifficulties were encountered inthe sampling and/or holding conditions of the samples. The relative percent difference (RPD) of duplicate samples or relative standard deviation (RSD) should be less than 15% for the precision of sample preparation and analysis to be considered in control. 9.4 Matrix Spike Matrix spikes are prepared for each sample type and analyzed to determine the matrix effect on the recovery efficiency. Matrix spike recoveries should fall within 25% of expected values. If the matrix spikes fail, evaluate the lab control spikes. If the LCS are within acceptance criteria there may be matrix issues in the samples. Discuss these inthe final report. ETS-8-154.1 Page 9 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 43 o f 122 E0S-0210 Interim Report #18 Northern Alabama Potable Water Systems, December2005 Matrix spike duplicates are prepared periodically to measure the precision associated with the analysis. Analyze a matrix spike and matrix spike duplicate (if prepared) inthe same run as the original sample. Matrix spike and matrix spike duplicate concentrations should fall inthe mid-range of the initial calibration curve or should be prepared at 1.5-5 times the endogenous concentration of the analyte. Spike concentrations should fall inthe low-range of the initial calibration curve if extremely low-levels are expected. Generally two or more levels are prepared, one in the low range of the curve and one in the mid-range. This avoids the need to pre-screen unknown samples prior to preparation. 9.5 Laboratory Control Spike Lab control spikes are prepared for each study to ensure recovery ofthe target analytes. These should be prepared at a minimum of 2 levels and in duplicate or triplicate. Recovery of these samples should be within 25% of expected values, and the RPD (or RSD) be 15%. If recoveries fall outside these limits the samples should be addressed in the final report. 10 Calibration and Standardization 10.1 Instrum ent Setup Note: In this example, a MicroMass UltimaTM Liquid Chromatography Tandem Mass Spectrometer (LC/MS/MS) is used. Other brands of LC/MS/MSs as well as single quadrupole mass spectrometers (LC/MS) may be used as long as the method criteria are met Brand names, suppliers, part numbers, and models are for illustrative purposes only. Equivalent performance may be achieved using apparatus and materials otherthan those specified here, but demonstration of equivalent performance that meets the requirements of this method is the responsibility of the laboratory. The operator must optimize and document the equipment and settings used. Establish the LC/MS/MS system and operating conditions equivalent to the following: Mass Spec: Micromass Ultima (Micromass) Interface: Electrospray (Micromass) Mode: Electrospray Negative, Multiple Response Monitoring (MRM) Harvard infusion pump (Harvard Instruments), fortuning Computer COMPAQ Professional Workstation AP200 Software: Windows NT, MassLynx 3.3 HPLC: Hewlett Packard (HP) Series 1100 HP Quaternary Pump HP Vacuum Degasser HP Autosampler HP Column Oven Note: A 4 x 10mm Hypercarb drop-in guard cartridge (Keystone, part # 844017 400) may be attached on-line after the purge valve and before the sample injector port to trap any residue contaminants that may be in the mobile phase and/or HPLC system. HPLC Column: Genesis C8 (Jones Chromatography), 2.1mm x 50mm, 4pm ETS-8-154.1 Page 10 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 44 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Column Temperature: 35C Injection Volume: 15pL Mobile Phase (A): 2mM Ammonium Acetate inASTM Type I water (See 7.3.1) Mobile Phase (B): Methanol Tim e. m in P ercen t M obile P ercen t M ob ile Phase A Phase B Flow R ate, m L/m in 0.0 60 40 0.3 0.4 60 40 0.3 1.0 10 90 0.3 7.0 10 7.5 0 90 0.3 100 0.3 9.0 0 9.5 60 100 0.4 40 0.4 13.5 60 40 0.4 1 4 .0 60 40 0.3 Note: Other HPLC gradients may be used as long as the method criteria are met. It may be necessary to adjustthe HPLC gradient in order to optimize instrument performance. Columns with different dimensions (e.g. 2.1mm x 30mm) and columns from different manufacturers (Keystone Betasil C18 etc.) may be used. Ions Monitored:__________________________________ A nalyte P rim a ry Ion P ro d u c t Ion A pproxim ate R eten tio n Tim e (m inutes) PFOA PFOS FOSA 413 499 498 169 99 78 5.0 5.2 5.8 Other product ions may be chosen at the discretion of the analyst, although m/z 99 is suggested for PFOS. Use of the suggested primary ion is recommended. Retention times may vary slightly, on a day-to-day basis, depending on the batch of mobile phase etc. Drift in retention times is acceptable within an analytical run, as long as the drift continues through the entire analysis and the standards are interspersed throughout the analytical run. 10.2 Tune File Parameters The following values are provided as an example. Actual values may vary from instrument to instrument. Also, these values may be changed from time to time in order to optimize for greatest sensitivity. A nalyte PFOA PFOS FOSA D w ell, sec 0.2 0.4 0.2 0.4 0.2 0.4 C ollision E n erg y, eV 10 25 30 60 20 50 Cone, V 20 30 50 80 30 60 ETS-8-154.1 Page 11 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 45 o f 122 ' E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Source Capillary Hexapole 1 Aperture 1 Hexapole 2 Source Block Temp. Desolvation Temp. A nalyzer LM Res 1 HM Res 1 Energy 1 Entrance Exit LM Res 2 HM Res 2 Energy 2 Multiplier G as Flow s Cone Gas Desolvation Pressures Gas Cell Set 2.6 3.5kV 0.5V 0.2V 0.8V 100-150C 250--400C Set 12.5 15.0V 12.5 15.0V 0.7V -2V 1V 11.0V 11.0V 1.0V 650V Set 150L/hr 700L/hr Set 3.0e-3mbar ETS-8-154.1 Page 12 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. 05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 46 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 10.3 Calibration Curve Analyze the standard curves priorto each set of samples. The validated method specifies that the standard curve should be plotted using a linear fit, weighted 1/x or unweighted. However, the standard curve may also be plotted by quadratic fit (y = ax5+ bx + c), weighted 1/x or unweighted, using suitable software. The calibration curves may include but should not be forced through zero. The mathematical method used to calculate the calibration curve should be applied consistently throughout a study. Any change should be thoroughly documented in the raw data. If the calibration curve does not meet acceptance criteria perform routine maintenance or prepare a new standard curve (if necessary) and reanalyze. For purposes of accuracywhen quantitating low levels of analyte, it may be necessary to use the low end of the calibration curve rather than the full range. For example, when attempting to quantitate approximately 50 pg/mL of analyte, generate a calibration curve consisting of the standards from 25 pg/mL to 1000 pg/mL rather than the full range of the curve (25 pg/mL to 25000 pg/mL). This will reduce inaccuracy attributed to linear regression weighting of high concentration standards. High and/or low points may be excluded from the calibration curves to provide a better fit over the linear range appropriate to the data or because they did not meet the pre-determined acceptance criteria. Low-level curve points should also be excluded if their area counts are not at least twice that of the method and/or solvent blanks. Any curve point may be rejected due to a bad injection or failing to meet accuracy requirements of 25% (and 30% for the LLOQ). Justification for exclusion of calibration curve points will be noted in the raw data. A minimum of 6 points will be used to construct the calibration curve. 10.4 Continuing Calibration Verification (CCV) Continuing calibration verifications (CCV) are analyzed to verify the accuracy of the calibration curve. Analyze a mid-range calibration standard, one of the same standards used to construct the calibration curve, at a minimum after every tenth sample, not including solvent blanks, with a minimum of one per sample set. Calibration verification injections must be within i25% to be considered acceptable. The calibration curve and the last passing CCV will then bracket acceptable samples. Multiple CCV levels may be used. 10.5 System Suitability A minimum ofthree system suitability samples will be injected at the beginning and end of each analytical run. Typically these samples are run priorto the calibration curve. The system suitability injections must have area counts with an RSD of 5% and a retention time RSD of S2% when evaluated independently. 11 Procedures 11.1 Extraction Scheme Allow samples to equilibrate to room temperature. Thoroughly mix samples by gently inverting the sample bottle. Measure 40mL of sample into 50mL polypropylene centrifugetubes (Spike the Matrix spikes as required*, replace lid and mix well). Note: * Samples may need to be prescreened to determine an appropriate matrix spike level (typically 50 150% of sample concentration). Alternatively the samples could be spiked at more than one level, allowing for the inappropriate spike level to be eliminated. ETS-8-154.1 Page 13 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. 05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 47 o f 122 E0S-0210 Interim Report #18 Northern Alabama Potable Water Systems, December2005 Condition the C18 SPE cartridges (1g, 6mL) by passing approximately 10mL methanol followed by approximately 50mL ASTM Type I water (flow rate approximately 2 drop/sec). Do not let column run dry. Note: Forthe following steps, maintain a -1 drop/sec flow rate. Do not allow the column to run dry at any time. Load the analytical sample onto the C18 SPE cartridge. Discard eluate. Ten mL of the 40% methanol inwater wash mixture is passed through the C18 SPE cartridge to rinse away potential interferences and then discarded. This step must be omitted if perfluorinated compounds with chain lengths less than C8 are targeted since these will be lost during this wash step. Elute with exactly 5mL of 100% methanol. Collect eluate into graduated 15mL polypropylene centrifuge tubes. This is the target elution fraction (final volume approximately4.5 mL as not all of the solventwill leave the SPE column. This will not affect the calculations in any way since the curve is also extracted). A n aly ze a portion o f the target elution fraction eluen t using negative electrospray HPLC/MS/MS or HPLC/MS. Note: Samples are concentrated by a factor of eight during the extraction; Initial Vol = 40mL -> Final Vol. = 5mL. Samples are stable at room temperature for at least 24 hours. Analytical samples may be stored in a refrigerator at 42C until analysis. S ta n d a rd iz a tio n o f C 1 8 S P E c o lu m n s --If poor recoveries a re observed, it m a y b e n ecessary to Standardize the C18 SPE columns in the following manner before analyzing samples. Use a standard with an analyte concentration between 1000 and 4000 pg/mL. Repeat the extraction scheme from the beginning up through the eluting with ~5mL 100% methanol. After the eluting with ~5mL 100% methanol step, collect an additional post-elution fraction by eluting with an additional 5mL of 100% methanol. Analyze both fractions by HPLC/MS/MS or HPLC/MS. If the target fraction contains a minimum of 85% of the respective analytes, it may be considered acceptable. If the wash contains significant standard (>15%), either the wash volume or percentage of MeOH should be decreased. If the post-elution fraction contains significant standard (>15%), the target elution volume should be increased. 11.2 Sample Analysis Set up analysis sample queue. Inject the same volume (between 5 25pL) of each standard, analytical sample and blank into the instrument. All samples with a concentration > ULOQ must be diluted and reanalyzed. If dilution of the final extract fails to produce acceptable results (e.g. poor MS recoveries) dilute the original sample and re-extract 12 Data Analysis and Calculations Calculate the analytical sample (extract) concentration from the standard curve using the following equation; ETS-8-154.1 Page 14 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. E0S-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 48 o f 122 E0S-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Extract Concentration, pg/mL = (Peak area - intercepts (slope) Calculate the percent recovery of the FSCS using the following equation: FSCS % rec. = (FSCS cone.. oo/mL) x 100 (Cone, added, pg/mL) Calculate the percent recovery of the MSs using the following equation: MS % rec. = (MS cone., pg/mL - Sample cone., pg/mL) x 100 (Cone, added, pg/mL) 13 Method Performance Note: Any method performance parameters that are not achieved must be considered in the evaluation ofthe data. Nonconformance to any specified parameters must be described and discussed in any reporting of the data. If criteria listed inthis method performance section are not met, maintenance may be performed on the system and samples reanalyzed, or other actions taken as determined by the analyst Document all actions in the raw data. If data are to be reported when performance criteria have not been met, the data must be footnoted on tables and discussed in the text of the report 13.1 System Suitability A minimum of three system suitability samples will be injected at the beginning and end of each analytical run. Typically these samples are run prior to the calibration curve. The system suitability injections must have area counts with an RSD of S5% and a retention time RSD of 52% when evaluated independently. 13.2 Quantitation Calibration Curve: The coefficient of determination (r2) value for the calibration curve must be greater than or equal to 0.990. Each point in the curve must be within 25% ofthe theoretical concentration with the exception ofthe LLOQ, which may be within 30%. Demonstration of Specificity: Specificity is demonstrated by chromatographic retention time (within 3% of standard) and the mass spectral response of unique ions. 13.3 Sensitivity Solvent Blanks and Method Blanks: Solvent and method blank area counts must be < 50% that of the lowest standard used in the calibration curve. Limits of Quantitation (LOQ): The lower LOQ (LLOQ) is the lowest non-zero active standard in the calibration curve; the peak area of the LLOQ must be at least 2X that of the extraction blank. By definition, the measured value of the LLOQ must be within 30% of the theoretical value. ETS-8-154.1 Page 15 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. E05-0210 Interim Report 918 Northern Alabama Potable Water Systems, December2005 Page 49 o f 122 E0S-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 13.4 Accuracy CCV Performance: Calibration verification injections must be within 25% to be considered acceptable. The calibration curve and the last passing CCV will then bracket acceptable samples. Multiple CCV levels may be used. Matrix Spikes: Matrix spike percent recoveries must be within 25% ofthe spiked concentration. If matrix effects are suspected, evaluate the LCS results to determine if a matrix effects are present and if the method is in control based on compliant LCS results. Discuss all results in the analytical report 13.5 Precision Reproducibility: Reproducibility of the method is defined by the results of duplicate or triplicate analysis of samples. A RPD or RSD of 15% will be considered acceptable. System Suitability: The system suitability injections must have area counts with an RSD of 5% and a retention time RSD of 2% when evaluated independently. 14 Pollution Prevention and Waste Management Sample extractwaste and flammable solvent is discarded in high BTU containers, and glass pipette waste is discarded in broken glass containers located in the laboratory. 15 Records Each data package generated for a study must have the following information included: study or project number, acquisition method, integration method, sample name, extraction date, dilution factor (if applicable), and analyst. Print the tune page, sample list, and acquisition method to include in the appropriate study folder. Copy these pages and tape into the instrument run log. Plot the calibration curves as described inthis method, then print these graphs and store inthe study folder. Print data integration summary, integration method, and chromatograms and store in the study folder. Summarize data using suitable software and store inthe study folder. 16 Attachments None. 17 References "Method ofAnalysis forthe Determination of Perfluorooctane sulfonate (PFOS), Perfluorooctane sulfonylamide (PFOSA), and Perfluorooctanoate (POAA) inWater", E. Wickremesinhe and J. Flaherty, Study Number 023 002, Centre Analytical Laboratories, Inc., State College, Pennsylvania, January 2000. Validation report for the "Method of Analysis forthe Determination of Perfluorooctane sulfonate (PFOS), Perfluorooctane sulfonylamide (PFOSA), and Perfluorooctanoate (POAA) inWater", E. Wickremesinhe and J. Flaherty, Study Number 023 002, Centre Analytical Laboratories, Inc., State College, Pennsylvania. ETS-8-154.1 Page 16 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 50 o f 122 E0S-0210 Interim Report #18 Northern Alabama Potable Water Systems, December2005 18 Affected Documents None. 19 Revisions Revision Number 1 Revision Number U p d a ted to the n e w form at. C h an g ed Title. Section 1: S tates the validation o f 3 analytes, rem oves reference to E P A d o c u m e n t th a t's n o lo n g e r a p p licab le. Section 2: Provided fo r the extraction o f m o re than the 3 validated analytes, allow s the use o f a L C /M S system , not only the L S /M S /M S previously m entioned. Section 3: R evised definitions for field m atrix spike, field control spike, LLO Q , m etho d blank, a n d M D L. Section 5: R e w o rd e d the interferences, ad d e d recom m endation to use disposable pipettes. Section 6: R ecategorized and p ared down. S e c tio n 7: C h a n g e d s to ra g e tim e to 6 m o n th s. A d d e d m o re calibration p o in ts to the table. Section 8:A dded statem ent addressing labeling requirem ents and spiking procedures. E xpanded section 8.8. Section 9: N e w Section Section 10: C h an g ed som e o f the p aram eters in the tables. A llow ed for use o f different instrum entation. A d ded inform ation from section 12 o f previous version, extensively revised. Section 11 (section 9 in previous version): Clarification o f w ash step, stated exact volum e o f eluate is 5 mL, revised standardization process, rem oved requirem ent to use L C /M S /M S . S ection 12 (section 13 in p revious version: n o ch an ges Section 13 (section 14 in previous version): E xtensively rewritten. Section 14 (section 15 in previous version): no changes Section 15 (section 1 6 in previous version): M in or ch ang es to recording requirem ents. S ection 16 (section 1 7 in p revious version): R e m o v e d attachm ent. Section 1 7 (section 18 in previous version): R em o ved reference to EPA docum ent that n o lon ger applied to this S O P . Section 18: N e w section. Revision Date 7 ETS-8-154.1 Page 17 of 17 Determination of Perfluorinated Compounds in Water Using SPE and LC/MS. EOS-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 51 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Attachment C: Protocoland Protocol A mendments E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 52 of 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: POOOl 131 STUDY PROTOCOL Study Title: AnaPlyersifsluoofrPoehrefxluanoreosublufotannaetesu(lPfoFnHaSte),(aPnFdBS), PFeMirsfhalu,moCmrloaaomlcsSta,enVreuesmgueltMfUaostnoiioannntgiet,oL(SrPCminF/aMgOllPSSMr)/MoiangmSrWamfmoaartletrhL,eivS3eoMril,aDSnededcSaimmtuearnllt, Exygen Protocol Number: POOOl131 Performing Laboratory: Exygen Research 3058 Research Drive State College, PA 16801 Phone: (814)272-1039 Sponsor Representative: Michael A. Santoro Director o f Regulatory Affairs 3M Building 0236-01-B- 10 St. Paul, MN 55144 Phone: (651) 733-6374 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page I oj 65 Page 53 o f 122 E05-0210 Interim Report It18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: P0001131 DISTRIBUTION: 1) Jaisimha Kesari, Study Director, Weston Solutions 2) John M. Flaherty, Principal Investigator, Exygen Research 3) Michael A. Santoro, Sponsor Representative, 3M Company 4) Exygen Research Quality Assurance Unit E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 2 o f65 Page 54 of 122 E0S-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: POOOl I3l PROTOCOL APPROVAL Study Title: Analysis o f Perfluorobutanesulfonate (PFBS), Perfluorohexanesulfonate FU(PissFihnH,gCSL)l,aCam/nMsd,SPV/eMergfSelutafootrirootonhc,etSa3mnMeasluDllMefocnaamtautmer M(aPloFLnOiivtSoe)rrsiinnagnWPdarStoemgrr,aaSlmloMil,aSmemdiaml eSnetr,um Exygen Protocol Number: POOOl 131 APPROVALS JWaiessitmonhaSolutions M3MichCaoeml pAa.rSfygntoro, Sponsor Representative Jnoh/n/ My .?Flaherty, rincipal Investigator Exygen Research REixcyhgaernd RAe.seG:rj ident, Facility Management Date -07 o r- i& f t Date Shaffer, Techni; ,ead, Quality Assurance Unit Research 7 Date E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page J o f 65 Page 55 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: PO0O0O01l I131l TABLE OF CONTENTS TITLE PAGE..................................................................................................................................................................' DISTRIBUTION............................................................................................................................................................2 PROTOCOL APPROVAL............................................................................................................................................3 TABLE OF CONTENTS..............................................................................................................................................4 INTRODUCTION..........................................................................................................................................................5 TEST MATERIALS......................................................................................................................................................5 OBJECTIVE...................................................................................................................................................................6 TESTING FACILITY................................................................................................................................................... 0 STUDY DIRECTOR......................................................................................................................................................7 SPONSOR REPRESENTATIVE..................................................................................................................................7 PRINCIPAL INVESTIGATOR....................................................................................................................................7 PROPOSED EXPERIMENTAL START AND TERMINATION DATES...........................................................7 IDENTIFICATION AND JUSTIFICATION OF THE TEST SYSTEM...............................................................8 SAMPLE PROCUREMENT, RECEIPT AND RETENTION................................................................................8 SAMPLE IDENTIFICATION.....................................................................................................................................9 ANALYTICAL PROCEDURE SUMMARY............................................................................................................9 VERIFICATION OF ANALYTICAL PROCEDURE..............................................................................................9 METHOD FOR CONTROL OF BIAS......................................................................................................................." STATISTICAL METHODS.........................................................................................................................................11 GLP STATEMENT.......................................................................................................................................................11 REPORT..........................................................................................................................................................................1! SAFETY AND HEALTH............................................................................................................................................. *2 AMENDMENTS TO PROTOCOL.............................................................................................................................13 DATA RECORD KEEPING........................................................................................................................................13 QUALITY ASSURANCE............................................................................................................................................14 RETENTION OF DATA AND ARCHIVING..........................................................................................................>4 APPENDIX I, ANALYTICAL METHODS...............................................................................................................1? E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 4 o f 65 Page 58 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: P0001131 INTRODUCTION T(PhFeBpSu)r,popseeroflfutohrioshsetxuadnyesisultfoonpaetrefor(mPFaHnaSl)ysiasnfdor ppeerrfflluuoorrooboucttaanneessuullffoonnaattee (PFOS) in water, soil, sediment, fish, clams, vegetation, small mammal livers and small mammal serum using LC/MS/MS for the 3M Decatur Monitoring Program. The study will be audited for compliance with EPA TSCA Good Laboratory Practice Standards 40 CFR 792 by the Quality Assurance Unit o f Exygen Research. TEST MATERIALS pTehrefluorotheesxtanesumlfaotnearitaels(PFHaSr)eand ppeerrfflluuoorrooobcuttaanneessuullffoonnaattee (PFO(PSF) BaSn)d, are all supplied by 3M. PFBCShemical Name: Perfluorobutanesulfonate Molecular Weight: 338 supplied as the potassium salt ^FSO /K *) Lot Number: 101 Purity: 96.7% Transitions Monitored: 299 -> 99 Structure: FF PFHMCShoelemciuclaalrNWameigeh: t:Pe4r3fl8uosruophpelxieadnaessutlhfoenpaotteassium salt (CFnSOj'K*) Lot Number: SE036 Purity: 98.6% TStrrauncstuitrioe:ns Monitored: 399 -> 80 FFF F FFF Page 5 of 65 E05-0210 Intrim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 57 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: P0001131 PFOCShemical Name: Perfluorooctanesulfonate Molecular Weight: 538 supplied as the potassium salt (CgF^SCVK*) LPuortiNtyu:m8b6e.9r:%217 TStrrauncstiutrioen: s Monitored: 499 --> 99 OBJECTIVE T(PhFeBpSu)r,popseeroflfutohrioshsetxuadnyesisultfoonpaetrefor(mPFaHnaSl)ysiasnfdor ppeerrfflluuoorrooboucttaanneessuullffoonnaattee (PFOS) in water, soil, sediment, fish, clams, vegetation, small mammal livers and small mammal serum for the 3M Decatur Monitoring Program using the current versions of the following Exygen analytical methods: V0001780: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS" V0001781: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS" V0001782: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Sediment by LC/MS/MS" V0001783: "Method of Analysis for the Determination of Perfluorooctanoic V0001784: "AAMcciieddth((PPodFFOOoAAf A)) iinnnaFVlyiessgihseatfanotrdiotCnhleabmDy esLtbeCry/mMLiCnSa//MMtioSSn"/MofSP"erfluorooctanoic V0001785: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Small Mammal Liver by LC/MS/MS" V0001786: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Small Mammal Serum by LC/MS/MS" TESTING FACILITY 3SPE0thxa5oyt8negeeRC:neo(Rs8lel1eeas4gre)eca,h2rP7cDhA2r-i11v06e38901 Page 6 o f65 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 58 o f 122 STUDY DIRECTOR Jaisimha Kesari P.E., DEE WW14ee0ss0ttoWCnheSesosttoleunrt,iWoPnAasy, 1In9c3.80 FPahxo:n(e6: 1(06)107)017-0714-031761 j .kesari@westonsolutions.com SPONSOR REPRESENTATIVE 3MMichCaoeml Apa.nSyantoro Director of Regulatory Affairs 3SMt. PBauuill,dMinNg 05253164-401-B-10 Phone: (651) 733-6374 PRINCIPAL INVESTIGATOR John M. Flaherty Exygen Research 3058 Research Drive State College, PA 16801 jPohhonn.fel:ah(8e1r4ty)@27e2x-y1g0e3n9.com EOS-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: P0001131 DATESPROPOSED EX PERIM ENTAL ST A R T AND TER M IN ATIO N OItcitsobperro0p1o,se2d00th4attothDeecaenmablyetric3a1l ,p2o0rt0i5o.n oTfhethaisctusatul deyxpbeerimcoenndtaulctsetdartfraonmd termination dates will be included in the final report. E0S-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 7 oj 65 Page 59 o f 122 E05-0210 Interim Report U18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: P0001131 IDENTIFICATION AND JUSTIFICATION OF THE TEST SYSTEM The following are the test systems for this study: Water (groundwater and surface water) Soil Sediment Fish Clams Vegetation Small Mammal Liver Small Mammal Serum bTehepusarmchpalseesdwainlldbperceoplalreecdtedbybythWe teessttoinngSofaluctiliiotnys.. PTuhrechcoansetroalndsapmrpolceessswinilgl wdeitthailtshifsosrtuthdey.control samples will be included in the final report associated The test systems were chosen to access the environmental impact of PFBS. PFHS and PFOS in the Decatur, Alabama area. SAMPLE PROCUREMENT, RECEIPT AND RETENTION Water, soil, sediment, fish, clam, vegetation, small mammal liver and small mammal serum samples will be received at Exygen directly from Weston Solutions. The details of sample procurement for this study are outlined in the 3M work plan entitled "Phase 2 Work Plan for Sampling Environmental Media." The number and types of samples collected will vary depending availability in the field. The total number of samples received and analyzed for each matrix will be documented in the final report associated with this study. Water, soil, and sediment samples will be used as received without further processing at Exygen. These samples will be stored refrigerated at 2C-8C. Fish, clam, vegetation and small mammal liver samples will be processed according to the appropriate analytical method (see Appendix I). These sfsraaammctppiolleenss wwwiiillllllbbbeeeceusnstetordriefudfogrefdrtohizneentshteuadtfiye<.ld-Sa1tm0tahClel. tmimSaemmoamfllaclomlslaeemrcutmimoanlwawnildlhotbhleee ssbteolroruoemdd frozen at < -10C. Trehpeorrteacnediprtaawnddaptaroacsesossciinagtedofwtihthe tshaemsptuledsy.will be documented in the final E05-0210 Interim Report tri8 Northern Alabama Potable Water Systems, December 2005 Page 8 o f 65 Page 60 o f 122 05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: P000113l SAMPLE IDENTIFICATION Prior to analysis, each sample will be assigned a laboratory sample reference lCnauhbmroorbmaetrao.troygTrsahapemhpricelefderatethanactwe iilsnl ubpmerboiecdreesnswetiidfllietdbherboyuungthihqoeuueltabatonhrdeatwaonriylallysdatiimcsatpilnlegpurrioesfcheerdeeuanrccehe. number. Sample storage conditions and locations will be documented throughout the study. ANALYTICAL PROCEDURE SUMMARY References: V0001780: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS" V0001781: V0001782: "Method of Analysis for the Determination of Perfluorooctanoic AA"Mcciieddth((PPodFFOOoAAf A)) iinnnaSlSyeosdiilsimbfyoenrLttChbe/yMDLSeC/tMe/MrSmS"i/nMatSio"n o f Perfluorooctanoic V0001783: V0001784: "Method o f Analysis for the Determination of Perfluorooctanoic A"Mciedth(PodFOofAA) innaFlyissihs afnord tChleamDestberymLiCna/MtioSn/MofSP"erfluorooctanoic Acid (PFOA) in Vegetation by LC/MS/MS" V0001785: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Small Mammal Liver by LC/MS/MS" V0001786: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Small Mammal Serum by LC/MS/MS" Tishome earbsoovfePFmBetSh,oPdFsHuSseandanPaFlyOtiSc.alTchoenfdinitaiol nrsepocratpwabillel inocflusdeepathraetiisnogmethres summed into total PFBS, total PFHS, and total PFOS found. VERIFICATION OF ANALYTICAL PROCEDURE A laboratory control sample will be used for the preparation o f fortified cmoentthroodl ,saanmdpaledsd.edTthoethteesmt asturbicsetasnvciea awmillicbroepmipaedttee.into solutions as per the bFbooottrttllweessa.tewrTislhlaembseeplib5no0gt0,tlEemsxyLhgaevpnerewbclieelelannsuerpdopulStyicnoie/nSlyepebucosettPdlreefpomerirefrslaumworipodlceehcemomlolieuccathtledsHa. mDTPphlEee Page 9 o f 65 05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 81 o f 122 E05-0210 Interim Report 918 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: POOOl 131 collection at the testing facility and have been shown to be free of PFBS, PFHS and PFOS. Samples will be added to each container to a volumetric fill line at 200 mL. A field duplicate, a low field spike and a high field spike of each sample will be collected. The low and high field spike bottles will contain PFBS, PFHS and PFOS as well as perfluorooctanoic acid (PFOA) and i1n.c2l-u1d3eCd ipnerthfleuosorlouotcitoannsouicsedactiodsp(li3kCe thPeFOsaAm)p. lesP. FTOhAe raesnudltsl3fCor PPFFOOAA aanrde l3C PFOA will not be reported in this study. Exygen will supply one field blank (control water) and two field blank spikes (control water fortified with cPoFllBeSct,ePdF. HSAatndthPeFOteSstinatgaflaocwilitayn,dehaicghh lweavteelr) fsoarmepvleery(etwxcelnutdyinsgamfpielelds duplicates and field spikes) will be extracted in duplicate and will also be fortified at a low processed through and high concentration the described procedure wtoitdhetPeFrmBiSn,ePmFeHthSodanadccPuFrOacSy aanndd to check for bias. For soil, sediment, clams, and vegetation, Exygen will supply one 500 mL parzeicpl-esaenaeldbSagc.i/SAplelccoPnrteaminieerrsw/bidaegsmuosuetdhfHorDsPaEmpbloettcleolpleecrtisoanmpwliellcboelleschtiepdpeodr tfoieltdh.e sAamt tphlee ltoecsatitniognf.acSilaimtyp, leeascwhilslabmepaleddweidlltobeeaecxhtrcaocntetdaininerdourpbliacgateinatnhde will also be fortified at a known concentration with PFBS, PFHS and PFOS at both a low and high level and processed through the described procedure to determine method accuracy and to check for bias. For small mammal liver, Exygen will supply a 50 mL polypropylene centrifuge tube. For small mammal serum, Exygen will supply a collection kit for each sample containing serum separator tubes (red top), vacutainers, needle holders and needles, transfer pipettes, and polypropylene tubes. At the testing facility, each liver and serum sample will be extracted in duplicate and bdwoeittlehlramalsilonoebwme afenothrdtoihfdiiegadhccalutervaaecklynaoanwnddnptcorooncchceeesncsektrdafottihrorbnoiauwsgi.hththPeFBdeSs,cPriFbHedS parnodcePdFuOreS taot Low and high spiking levels for each matrix are defined below: MWaattreirx Low Spiking Level High Spiking Level 500 ng/L 5000 ng/L Soil 4ng/g 40 ng/g Sediment Fish 140nngg//gg 40 ng/g 100 ng/g Clams Vegetation 10 ng/g 10 ng/g 100 ng/g 100 ng/g Small Mammal Liver 10 ng/g 100 ng/g Small Mammal Serum 10 ng/mL 100 ng/mL E05-0210Interim Report 818 Northern Alabama Potable Water Systems, December 2005 Page 0 o f65 Page 62 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: P0001131 Rleevceolsv;erhioews eavreer,atnhteiciepxaatcetdptroecbiseionbeatwndeeancc7u0r%acyawndill1b3e0%detoerfmthineedfobrtyiftiehde aacncaulyrsaicsy owfillthbee iqnucalulidtyedcoinnttrhoel fisnaaml prelpesortd.escribed above. A statement of METHOD FOR CONTROL OF BIAS hCoomntorgoelnoefobuisasmwixitllurbee oafddearcehssemdabtryixtafkroinmg ruenptrreeasetendtactoivnetrsoulbs-asammppleles,s afnrodmbya analyzing at least two levels of fortifications. STATISTICAL METHODS cSatalctuisltaitcisonwoilfl abveerliamgeiterdectoovetrhioesse, asspaecpipfileicdabinle.the subject methods and to the GLP STATEMENT All aspects o f this study shall be performed and reported in compliance with EPA TSCA Good Laboratory Practice Standards 40 CFR 792. The final rthepeorsttuodrydawtaaspaccoknadguect(esudppinliecdomtoptlhiaenScpeonwsiothr) csuhrarllenctonatnadin aapsptlaitceambleentGthLaPt standards and will outline any deviations in the study from those standards. This statement will be signed by the Study Director and Sponsor Representative. REPORT Aat tfhineaclornecplourstiownilol fbtehpersetpuadrye.dTbhyethreepporrtinwciipllalinicnlvuedseti,gbautot rwoilrl tnhoetirbedelsimiginteede to, the following: The name and address of the Study Director, Sponsor Representative, and of the testing facility. A statement of GLP compliance (any related documentation, such as chain-of-custody records, must be in the study records). E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page I / o f 65 Page 63 o f 122 E05-0210 Interim Report #18 Northern Alebama Potable Water Systems, December 2005 Exygen Protocol Number: POOOl 131 The signed and dated statement by the Exygen Research Quality Assurance Unit regarding dates o f study inspections and dates findings were reported to the Study Director and Management. tAchoepdyesusocbrfjieptchttieomnaenotahfloytdthiecwaelaxsmacfeottlhaloondwa.leydtAicenaxylaccmotlnoydd, iiifttiicoiasntsnioeencmsepstsloaorytyheditsoimninetcthhleuoddsteuwodinyll.lybIeaf incorporated into the report. If the method is photo-reduced, the project number and page number must be included on each page. Description of the instrumentation used and operating conditions. All results from all sets analyzed. Control and fortified samples will be identified and the data table will include sample number and fortification level. Rcaherfpoorrmetsiafeitncoatgatritaoivmneslceohvfreoal.mstaaTtnohdgaerradlmoacsnaftdoioranceoaoncfhtrtohalnesaaalymntaeplylient,eaepnaedcahakmschawrtorilimlx,abtieongccrlulaemdairnlaygt identified in all chromatograms. All circumstances that may have affected the quality or integrity o f the data will be documented in the report. Locations where raw data and the final report are to be archived. Additions or corrections to the final report shall be in the form of an amendment signed by the Study Director. The amendment shall clearly identify that part of the report that is being altered and the reasons for the alterations. The amendment will be signed and dated by the Study Director and the Sponsor Representative. A79ll2.a1p8p5l.icable requirements for reporting o f study results as per 40 CFR SAFETY AND HEALTH Laboratory personnel will practice good sanitation and health habits. Every reasonable precaution shall be taken to prevent inadvertent exposure of personnel and the environment to the test or reference substance(s). E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 12 o f 65 Page 84 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: P0001131 AMENDMENTS TO PROTOCOL Aexlpl ressigsendificinanwt ricthinagn,gesisgnteod tahned adnaatleydticbayl tphreotSotcuodlyoDutirliencetdor haenrde Swpiollnsbore Representative. Amendments usually will be issued prior to initiation of study plan change. However, when a change is required without sufficient time for the issue of a written amendment, that change may be effected verbally with supporting documentation signed and dated by the Study Director and followed with a written amendment as soon as possible. In this case, the effective date of the written amendment will be the date of the documented scthuadnygep. laCnopcoiepsieosf. theThsiegnoerdigainmaelnadmmeenndtsmweniltl bweilal pbpeenmdeadinttoaianlelddiswtritihbuttehde original study plan. Any deviations from the study plan or from the analytical method as provided will be documented and reported promptly to the Sponsor Representative. DATA RECORD KEEPING Records to be maintained include the following (as appropriate): Sample tracking sheet(s) Sample receipt records, storage history, and chains of custody History and preparation of standards (stock, fortification, calibration) Description of any modifications to the method Instrument run sheets, bench-sheets or logs AAnllaclyhtriocmaladtoagtaratpabhliecsand instrumental conditions SAacmopmlepleextteralicsttiionng aonfdstaundaylypseisrsdoantneesl, signatures and initials Chronological presentation of all study correspondence Any other documentation necessary for the reconstruction of the study Chromatograms- All chromatograms will contain the following: Sample identification, injection date, arrow or other indication of the area of interest, and injection number corresponding to the run. Additionally, fortifications will include the amount of analyte added and the sample number o f the sample that was fortified. Analytical standard chromatograms will additionally include the concentration (e.g., pg/mL). Page 13 o f 65 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 65 of 122 EOS-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: POOO1131 Aansaplyatritcaolf sthete: daorcuunmsehnetaettiolinsttihneg ftohleloswaminpgleshs eteotsbwe irlul nbeinintchleudseedt, iannedacahn instrument conditions sheet describing the instrument type and operating conditions. QUALITY ASSURANCE The QA Unit of Exygen Research will inspect the study at intervals adequate tSotuadssyuDreirceoctmorp,liEaxnycgeewnitMhaGnaLgPe'ms,eannt,dawndillthreepSoprotnthsoerfRinedpinregssenotfaatiuvdei.ts to the RETENTION OF DATA AND ARCHIVING All hard copy raw data, including, but not limited to, the original wchitrhomthaetodgartaampsa,ckwaogrekssuhbeemtsit,tecdortoretshpeonSdtuednyceD, iraencdtorre. suTlhtsessehwalilllbbee ainrcclhuidveedd with the original study plan, amendments, final report, and all pertinent information from the Sponsor. The testing facility shall keep all electronic raw data and any instrument, equipment, and storage logs for the period of time specified in 40 CFR 792.195. An exact copy of the materials submitted to the study director will also be kept at Exygen Research. Exygen will obtain permission from the study director before discarding or returning samples. E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 14 o f 65 Page 68 of 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: POOOl I3l APPENDIX I ANALYTICAL METHODS V0001780: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS" V0001781: V0001782: V0001783: "Method of Analysis for the Determination of Perfluorooctanoic "AA"MMcciieeddtthh((PPooddFFOOooAAff AA)) ininnnaaSlSlyyeossdiiilssimbffyooenrrLtttChhbee/yMDDLSeeC/ttMee/MrrSmmS"ii/nnMaattSiioo"nn of of Perfluorooctanoic Perfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS" V0001784: "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Vegetation by LC/MS/MS" V0001785: V0001786: "Method of Analysis for the Determination of Perfluorooctanoic A"AMcciieddth((PPodFFOOoAAf A)) iinnnaSSlymmsiaasllllfoMMraathmmemmDaaelltLSereivmrueirmnbabtyiyoLnLCCo/Mf/MPSeS/rM/fMlSuS"o"rooctanoic 054)210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 15 o f 65 Page 67 o f 122 E05-0210 Interim Report 818 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: P000I I3l AMNeAthLodYNTuImCbAeLr: MV0E0T01H7O80D Method of Analysts for the DeterminbaytLioCn/MofSP/eMrfSluorooctanoic Acid (PFOA) in Water Analytical Testing Facility: E3S0txa5yt8egeRCnoeRslleeeasgreeca,hrPcDAhriv16e801 Approved By: TPeauchl nPCinocnalnoLlelayvder. LC-MS, Exygen Research io I Date JVoihcne PFrleashiedretynt, Op/erations, Exygen Research Date Total Pages: 7 E05-0210 Interim Report U18 Northern Alabama Potable Water Systems, December 2005 Page 16 o f 65 Page 68 o f 122 E0S-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: P0001131 Exygen Research Method Number V000I780 analytical m ethod Method of Analysis for the DeterminatiLoCn/oMfSP/eMrfSluorooctanoic Acid (PFOA) in Water by 1.0 Scope TaScphieidsctmbroyemthHeotirdgichisDtPeoetebrfceotoremrm(apLnlCocey/MedLSif/qMouriStdh)eiCnihsworoalamtetirao.tnogarnadphqyuancotiutaptlieodn otof paerftlaunodroemoctaMnoasics 2.0 Safety 22..12 pACrolewncsaauuyltstioothnbess.earpvperosapfreialtaebMorSatDorSybperfaocrteicheas.ndling any chemical for proper safety 3.0 Sample Requirement 333...132 33..54 3.6 tNmSAASAeaamlnotimlmnylpsseuppaaeatsllmemrsaeeastmspptc4uallpo0enerslelsdetplm.oesmrtcrLohcetuceidoosoenstnfsutbsrtapeeipeinefsnrrtgrtoihinngscioaagseertmdraonpaunuepatrteeglreeddtuhsieslcfydeowlsedrhmifsloeoflixsoruxhbtrlwerdoetadhaucstebltbepdieoreeefxnbscota.eirrafmeailcelcpbeodtleinweoitisnnern.idg.futshgateoemdspaealmeqtdup-i3lfliio0nbrg0rea0pxteltrarpanmtcotfioofornror.toh-mi5s project. 4.0 Reagents and Standards 4444....2314 MAPWemeartftmhleuaroon-nroioHluom-PctLHaACnPcoLgeiCtcraaAtdgeerc-aiddAe-.CS.iSg.mRae-aAglednritcGhrade 3.0 Instrument and Equipment 5.1 555555......342567 A5ADsA1p1o05L2nilds5vammhpcel-eioyovmLLgnntsihticLaddnscebiieaLspsellctpepDqeotbooeurmPcafsdsiooplEaaiartclbpbonlmnrelleoceeadacaeprtppnrtiwcaoorocpaanllewieyypwtdth-sppaelmdbirrm(aqoo5laoeuppt0vuMaiyyo-adt1llhfrafeeoiOsranbncresOboheeapluStrdeccetLopleeiae,mvnennks1cogtt.atrri0ltnrtiiuo0oofftmuu-gem20ggrge0a.eeer00pati0ettnuthuu0rijLobbe0(cen)eLc1.asstCao..pgnr/a.MdbclaqSepu/aMaobnfSltei)tp.aoutfimoinnp.jiencgtinugp 5t-o2002 5555....189110 2DAWmiuastptLoeoprcssialpeSbeaeltretpeHpsPiP(ap1Lke0tCVt0e-avs1.ci0a6l00kciept.L(lga)ndtC1108-1S0P0EpcLa)r,trwidigthesd.isposable tips. Page 2of 7 Report #18 Northern Alabama Potable Water Systems, December 2005 Page 17 o f 65 Page 58 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: POOOl 131 Exygen Research Method Number VOOO1780 ANALYTICAL m e t h o d _________________________ Method of Analysis for the DeterminatiLoCn/oMfSP/eMrfSluorooctanoic Acid (PFOA) in Water b'v 55..1132 CSPenEtrviafucgueumcapmaabnliefoolfds.pinning 50 mL polypropylene tubes at 3000 rpm. 6.0 Chromatographic System 6.1 (APn/Nal:yt8i2ca5l0C5-o0l5u2m1n3:0F)luophase RP (Keystone Scientific), 2.1 mm x 50 mm. Su 6666....3245 TMMGeroomabbdpiiilleeeenraPPtthhPuaarreoss:eegr((3aABm0))":C:: M2 eotihManAolmmonium Acetate in Water Tim22e08l02...O..(0005min) %2666255555A %7333755555B Flow Rate (mV00000.....m33333 in) 666...867 RIQnuujenacnTttiiiotmanteiV:ono-:lu2mP3eeam:kin1Au5rteepasL.-(ceaxntebrneailnsctraenadseadrdtocaalsibmrauticohnacsu5rv0ep. L). The above conditions are intended as a guide and may be changed in order to optimize the HPLC system. 7.0 MS/MS System 7.1 Mode: Electrospray Negative MRM mode, monitoring 413- 369 m/z. TophteimaibzoevtehecMonSdMitiSonssystaerme .intended as a guide and may be changed in order to 8.0 Preparation of Solutions 8.1 Mobile Phase 8.1.1 a2mmmMonaiummmaocneituamte taoce1t0a0te0 imnLwoaftewraitserp.repared by adding 0.154 g of Alternate volumes may be prepared. Page t u - E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 18 o f 65 Page 70 of 122 EOS-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 E xygen Protocol Num ber: P0001131 Exygen Research Method Number V0001780 _________________________ ANALYTICAL METHOD_________________________ Method of Analysis for the DetetminatiLoCn/oMfSP/eMrfSluorooctanoic Acid (PFOA) in Water by 9.0 Standard Preparation 9.1 999999S......t111111a......n156342darmTmaAPAAimomafam1Adnrp0rhfeoLLL1e1Sep00at1e1ma2t2hptr.m.h.n0oo1oo330oa10saaatntc1ff2Lfxrpmhomnpplekot5titygeohchggmhl/pLLtoa/Fklmee//eiidgmmmacnofisLLaa/L1tna1mLr1tetnLLatDD.olt0e0h0laiLdLcysff0PP1effopoikDptoo1f2cEEa4gfrpgof2rr50Pantosp/ttgr/5bibtmioidCEm.rtfmri/ff-oo1itoielamiimfiLcuttLccbLprnfiattactpdiaaLolLlaniscteeaSsttLgortiodaii(..ottoaLsooon/DcliltmltoneouilannoDuiouoP.lontrrtLusnitssPeErfinoiosotooeoE.~ionsslllcbnosnuotuu1lotobantutetltt0oltiobouiitduto0ootitlltoaotntaeeniniflopn.ooefafooofngsin.ironnfff/fapromaiaPPPrtnlulfaoeFFFLarvvPlOOiOmtotoaooFyvAAlAlaOfuu)boxfmmPeliiaAiiiunsssmFnesemadpOpptiluoosorrremAdereeffvpexopippo1lrd1abaapufel00rrruyetpi01ee0eenrma0dddiwwwrota0eeobbbediidrwttiyyyehohg1ofib0hbbbftrfmmyh0rriirgnii6iee1nnnmbmegtt0hghggmrr0eLaaaiiii1ntnnnnnoth0ogwggwoognarillmtniinh111ttaoii1hgh1gn0nst1l1 9.2 Standard Calibration Solutions 9.2.1 9.2.2 pciTLdarhCeeleinp/bMtafriorcaSealtild/looMtanwoSsissnntacgeamaenliddpisbelaerdarasd..ttsiyopanircesatlparneoxdcaaemrsdspseleda:retahdrpodrueitpgioahnreathdlecionenxHctreaPncLttrCiaotniwopnarstoermc.eadyuTrbheee, oSCfooFlnuoctrie111to111ni0000f0n00t000irc(aanttioioobnn) Fortification Volume (442211P0000000L000000) ForVtifoileudmCeoonftrol Samp4444444le0000000 (mL) CSotCanncadelF1ina521b025irt050n0rrd500a00a0a0t(tlipioopnnt)o*f XXXXXXXSCCCCCCCC(teaammmmmmmxlnimmmmammmdbmardddddddardpdddddddtiyylvyyyyoeyIyvyyyyn)D*------5623401 * cTohneceexnttrraactitoend,cdounecetonttrhaeticoonnocfenthtreactiaolniborafttihoenssttaannddaarrdd disueriqnugatlhteoe8xxtriatcstiinointia(Sl PE) XC " extracted calibration standard. Page 4 of Report #18 Northern Alabama Potable Water Systems, December 2005 Page 9 o f 65 Page 71 of 122 EOS-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: POOOl 131 Exygen Research Method Number V0001780 ________________________ ANALYTICAL METHOP________________________ Method of Analysis for the DeteiminatiLoCn/oMfSP/eMrfSluorooctanoic Acid (PFOA) in Water by 9.2.3 9.2.4 9.2.5 nsAaSAeettltoe2tzdeore"ereCrfndoasa.ttltoalsentea6vdxnoatCdrrlauad,crmsudtpeeedxsstotocralaauntcwldtitibeoocdrnoaw.nt(iecroeeenkansgst.retaantntidobnalrsadnoskfi)nstma1nu5ds-mtarbLdesppmorlaeyypparbroeepdpyrlweenpiteahrteuedabceahss 10.0 Batch Set Up 10.1 10.2 cRopiEnonraeonetcqhcthureeroidebrqlueasaurmgtaacefllehnoinrttryetotscicfafoifosesovsnaduertmrrrfyoaiapenltlflceod(eskrmanptehneolxatedwhtnrbolnaaafdcotbtccreohbodtr.lhanati(cnsoteykrpnyprtiourdcajsauetiilcnpoltylgn.ics2aH0te(PlsoaLrbaCnledcwsossna)pttiremkore)lussatswnpiidinlklceltb)uwedotesopraeetvcaeilgferieaietnsdytt 11.0 Sample Extraction 11.1 11.2 1111..34 11.5 AMmfLCEi1no5olonitleuxolaanmaotdledswSwyLuisOztweeraipeleodmmlios)nt4bh.lapLy0ymtlp~hepmSpre5ooolmLlnepCymsyLcpotLlsuorfeoosnnsSipfd1anePyHi0mgtcEl0iPeeope%nnLlncleeetaCercmrdiocttfwrerreuCointgaadshtttesgprapeiernSrtofausuoPr(ybgtl~(Ei.e1Leo2snCctgC(uad/fo,rMbroitonfer6llSpaissedla//mcgs(MmvfeteLoocSp.Srl))l.ut.eDibmmfDdyiysLeioclapu=asonatre5sdofnsdteemeienlltleuudoLgtaae)4c.tdt1eeo0,0.lrumienmmpLtoLlnawcgrmieurtanheldtidhduwraayaanttenoeddlr 12.0 Chromatography 12.1 LInCje/Mct Sth/Me sSamsyesatemmo.unAt ocfaelaibcrhatsitoanndsatradn,dsaarmdpmleuasntd pfroerctiefdieed asnamd pfleollionwto tahlel 12.2 12.3 bmieAaSfnoxentntualgtaelrnsliorayntedswczbpnnateeeieetrnidddrdrisgsensebcacodyasamlfunelbPiepddtbexFletretowdOaarsfatt.eiAicnoeetthnnexcaedotnersravactreaceneanntsrladeipydlbdyoaro5trnaicd-fcdt1asiaioal0nlinmbgsssreaastaatomty.mtaiopnapbtndlleeealsers.aidstnsaesjtAnte.ifdcsniatvteareeEddnrsoxspartareltmmatrecsutortehnserdeteadtecibbvsvoeeteenag,rnciiynanednnnca5ltiru-rned1adngst0etiiodrmosenafuamlssateetpvttlesbheoloesesf c(taolibacractoiounntsftaonr daarsdescomnudstsebteotfheexfitrrastctaendd sltaasntdianrjdecs)ti.onInineaithsearmcpalsees,eet.xtracted 12.4 Ugesneerlainteedarfosrttahnedaanrdalyctuerbvyeslinfoearrqreugarnetsitsaitoinonu.singLi1n/exawr esitgahntdinargdofcupervaeksaraerae Page 5 oi ^ Northern Alabama Potable Water Systems, December 2005 Page 20 o f 65 Page 72 of 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: POOOl 131 Exygen Research Method Number V0001780 [ _________________________ ANALYTICAL METHOD_________________________ Method ofAnalysis for the DeterminatLioCn/oMfPSe/MrflSuorooctanoic Acid (PFOA) in Water by 12.3 evsSoxeafcrmtsewupesadlercesatraseliynsbspdrtoaaerntmidso.ernessshptaoonundlsdaersdnsochtooneuxcldecnebeterdaftuisotratnhneudrsadirndiglurtMeesdapsoasnnLsdyernse.xan3Aa.l3nyyz(oedsra.emqpuilevsalethnatt) 13.0 Acceptance Criteria 13.1 oCfh4ro1m3 aatmogur.amThmeu4s1t 3shaomwuappaereanktocfoarrdesapuognhdtesrtioonthaet P3F6O9 Aamauniforno,mwahiplaerethnet daughter ion (369 amu) represents the loss of carbon dioxide. 13.2 bmMlauensthtkobcdeoonbbtlaatainniknssePdmFauOnsAdt tnahotetleecnvotenilrtseaigsnreetPamFteOursAtthbaaent rlSeeO-veenxlgtsrfaLgcr,teetadht.eenr tahnanewthbelaLnOkOsamIpIlea 13.3 13.4 wtARe1hn3aenet0criyirr%roaevckneantsselreohiidtbewo.srouanloftdivfosaancblmouesneptstealr.evnosdalIlasfsurhpadaoitkeucfedooldsnubtnabryndoedlttrhosmepe-baiektexarentiraxafalacssylttplsaesitdtkio.setuotsitcAsmdaindeluytesoetrutmmhbtleaeiitenrabreicxecbtiewysfppeturiaeeksnb-ieenle7xgo0tulr-tiathm1sce3itidi0toseH%,nu7tgho0ieesf HErorworevTeers,t,thme atyotableneuxmclbuedredoffreoxmtratchteedccalacluiblraatitoionnosftatnhdearcdaslibtrhaattiocnoucludrvbee excluded must not exceed 20% of the total number of extracted standards 13.5 Tinhjeectceodr.relation coefficient (R) for calibration curves generated must be a0pp.9r9o2pri(aRteJ st0e.p9s85m).ustIfbecaltiabkreantiotno raedsujultsst finalsltruomutesnidteotpheersaetiolnim, iatsn,dthtehne standards or the relevant set of samples should be reanalyzed. 13.6 Retention times between standards and samples must not drift more than 4 % within an analytical run. If retention time drift exceeds this limit within an analytical run then the set must be reanalyzed. 14.0 Calculations 14.1 bgUaessneeedtrhaeotendfopblleyoawtkhienargMeaea)qsusuasLtiiynongnxtthsooefctsawtlacanurdelaaptrerdotghcruearmavme: o(luinnteaorfrPeFgOreAssifoonunpdar(aimn entge/rLs). PFOA found (ng/L) - fPeak aresalo-pientercept) x DF DF - factor by which the final volume was diluted, if necessary. Page t>oi 7 Report #18 Northern Alabama Potable Water Systems, December 2005 Page 21 o f 65 Page 73 o f 122 E0S-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: POOOl 13 1 Exygen Research Method Number VOOOl780 | ANALYTICAL METHOD | Method of Analysis for the DeterminatiLoCn/oMfSP/eMrfSluorooctanoic Acid (PFOA) in Water bv 14.2 tFhoerfsoallmowpliensgfeoqrtuiafiteiodnwtoithcaklcnuolwatne tahmeopuenrctsenotfrePcFoOveAryp. rior to extraction, use Recovery (%) = [total analyte found (ng/L) - analyte found in control (ng/L)] analyteadded (ng/L) Page 7 of 7 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 22 oj 65 Page 74 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: POOOl 131 AMNeAthLodYNTuImCbAeLr: MV0E0T01H7O81D Method of Analysis for the DeterminaLtiCon/MoSf /PMerSfluorooctanolc Acid (PFOA) in Soil b'y Analytical Testing Facility: ES3x0tay5tg8eeCRnoeRslleeesagereca,hrcPDhAri1v6e801 C--tLApproved By: TPaeuchl nCiocanlnLolelayder, LC-M$~, Exy'gen Research DateiqIu . / oJ Date Total Pages: 7 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 23 o f 65 Page 75 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number. P000113l Exygen Research Method Number VOOOI781 ANALYTICAL METHOD Method of Analysis for the DeterminaLtioCn/MoSf P/MerSfluorooctanoic Acid (PFOA) in Soil by 1.0 Scope TaScphieidsctmbroyemthHeotirdgichisDtPoeetebrfceotoremrm(apLnlCocey/MedLSif/qMouriStdh)eiCnihssorooliaml.tiaotnogarnadphqyuanctoiutaptlieodn otof paerftlaunodroemoctaMnoaiscs 2.0 Safety 22..12 AColwnsauylst othbesearpvperosapfreialtaebMorSatDoSrybperfaocrteicheas.ndling any chemical for proper safety precautions. 3.0 Sample Requirement 333...132 33..54 StANSAeaamltomlmlpsseppaaeallmmrseeatspptcu1lloe5erselstplg.oemrrcooetucfdisoettesnsbsretipenfsrHgtaohmgcioseerpdronlaueuetreegfeddohserlydwsehxmfilootlriruxablscedeotdiioslbbnpsee.eafcomirafpelilleebodsew.iinendgtshateomspaelmqedupillfiionbrgraeptxeltarantcotfioorrno.tohmis project. 4.0 Reagents and Standards 4444....1342 WMAPemearfttmhleuaroon-nrooiHulo-mPctLHaACnPcoLgeictCraaAtdgeecr-aiddAe-.CS.iSg.mRae-aAglednritcGhrade 5.0 Instrument and Equipment 5.1 555555555555............5176823111941230 W2ADAASU5ADs1p1o05PL2nluimiladtss5EvtmamrhtppceLo-aeleiomoovyvpLLsgrnncsisostaihtLciapaldndscncSeebbeieuiaiLaesespcltlctulertpepqDpoetembboeouHrpmsPPcaafsdsiioPo(tpElaamapiha1tcrlLbpkbeoln.m0arneleltCoVcent0eaadcaepeir-tpapvnsftr1iwaocc.roopoci0aanaleweill6d0wlytpydth.0s-kppaeclmdbirmcir(aptqoo5al.o(Leuppv0tuMlaiyay-ogtda1lrhlfaf)neeio0sardbnnctr0sebCoeheupa1lStr1eLdecct0op8leaeit-evmennkn1sSco1gt0t.atPir0rltr0nuitio0Eoofftmuu-pgem20cgggLre0aa.eeer0)r0ptai,e0tttnuthruuwir0jiLobbed0(in)ceecLgt1.hsasteCao..psgdnr/a..MidbcsplaqSepou/aMsaoabnbSflteil)te.paotutifimopinsnp..jiencgtinugp 5t-o2002 Page 2 oi'7 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 24 oj 65 Page 78 of 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: POOOl 131 IExygen Research ANALYTICAL METHOD Method Number VOOOl781 Method of Analysis for the DeterminaLtiCon/MoSfP/MerSfluorooctanoic Acid (PFOA) in Soil by 55..1145 WCernistrt-ifaucgtieocnasphaabkleer.of spinning SOmL polypropylene tubes at $000 rpm. 6.0 Chromatographic System 6.1 6666....2354 TA(MMGPenr/oomaNabbdlp:iiyilleetee8irn2caPPta5thhPul0aarrC5eoss-eeo:g0lr((5u3aAB2mm0)1)n:3C:::0F) lM2uoempthMhaansAoelmRmP o(KnieuymstoAnceetSactieeinntiWfica)t,e2r.1 mm x SOmm. 5p Tim2208el02...O.(0.050rpip) %6662255555A %3773355555B F(mlolw00000V.....33333mRiante) 666...876 QRInuujenacnTttiiiotmanteiVo: no~:lu2Pm3eeam:kin1Au5rtepeasL.-(ceaxntebrneailnsctraenadsaerddtcoaalisbmrauticohnacsu5rv0ep. L). TophteimabizoevethceoHndPiLtiConssyastreemin.tended as a guide and may be changed in order to 7.0 MS/MS System 7.1 PMFoOdAe:. Electrospray Negative MRM mode, monitoring 413 -- 369 m/z lor TophteimabizoevethceoMndSiMtioSnssyasreteimnt.ended as a guide and may be changed in order to 8.0 Preparation of Solutions 8.1 Mobile Phase 8.1.1 a2mmmMonaiummmaocneituamte taoce1t0a0te0imn Lwoaftewr aitserp.repared by adding 0.1S4 g of Alternate volumes may be prepared. Page J u f ' E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 25 of 65 Page 77 of 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: P0001131 Exygen Research Method Number VOOO1781 ANALYTICAL METHOD_________________________ Method ofAnalysis for the DeterminaLtiCon/MoSf /PMerSfluorooctanoic Acid (PFOA) in Soil by 1 9.0 Standard Preparation 9.1 9999S....t1111a....n1432darmaAmaPommAAidnrfL0L1eL1Sea11at22.pth1.0nooo55o0a1aacfff2rpmmnplpke(t5ytgoghhg/hLLta/Fleme//iemmmcoiLLs1Lna1Lr1tLLDDl.ot000aiLcsfPfP0fpfioptkD1ooEEcagrgp2rranPts/tt/g5ibbmtiodimEfiff-/ooiolaimimLcuttcLcbrnttadtaalloLLisetesSttotioi.i(.otooLonsoclllnoeulnnoDuuo.ltrtsuitssPrifooiootoeoE-inllcnloun1uutbntte0tttoiooiido0oottaontannflpeofofaoogi.rinfnffffparPiaPPnnlulFFFLarvvOOilOotooyvAlAAluf)uomPmlaiiiusssFneempdOppoorerrAedeeffpopip1l1aabafu00rrryt0e10eeed0ddwwwt0obbbeiittwyyiyhh1gi0bhbbtmmh0rrriniiiecnnnmgttmhhgggLeaaiii1ntnnnnh0ouggwgallmni111toiinhng000l 9.1.5 9.1.6 ATmaf1rp0hoep0temhrm.0osatt1nLxhooiecmlpokdiganfaa/tmtenatehldLy1eof24ffo50opr.Crtrm1itefipLfiacipancaLgrdata/DittmoiaioPonrLnenEs.sosbsotlooaullbtututiltloteeiion.ofnsnoaorrtfaoePmtFaoOabxfAeiinmsatiluosmrvpeordpeluepinrmairoaeeddreoofbfrfyig61be0mrr0ianotogwnritinhtahgst 9.2 Standard Calibration Solutions 9.2.1 9.2.2 TipLcdrahCeelenip/bMtafirrcoaSealtdli/looMtawnosSissnntacgeamaenlidpdisbelaedrarasd..ttsyiopanircesatlparneoxdcaaemrsdspseleda:retahdrpodrueitpgiohanreathdl ecionenxHctreaPncLttriCaotniwonparstoemcr eadyuTrbheee, * cXoTSCofhoConFlenuco-cteerie1t1txno1eiioni000ootfn0txot00rirrctaafarapcttatiiptiocoeobntndn)e,dcdocuFnaeoclVriettbo(oi4n22411pfrl00000ti00autLcr0h00000tmaa)iettoeiioncononsntoacFnfeoSdtnrhVaattiermrfoadilcpet.u4444444iadlome0000000lCni(ebmooronaLffttr)tioholen CSotaCnncadelF1ina251b205irt5000nrd5r00000aaa0(ttlipioopnnt)o* f ssttaannddaarrdd disueriqnugatlhteo XXXXXXXSCCCCCCCC(teaammmmmmmxlnimmmmammmdbmardddddddardpdddddddtyilyyyyyyoeyyyyIyyy)nD*------324560l e8xxtriatcstiinointia(Sl PE). Page 4 of 7 Report #18 Northern Alabama Potable Water Systems, December 2005 Page 26 o f 65 Page 78 o f 122 E0S-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: P0001131 Exygen Research Method Number V0001781 | ANALYTICAL METHOD Method of Analysis for the DeterminaLtiCon/MoSfP/MerSfluorooctanoic Acid (PFOA) in Soil by 9.2.3 9.2.4 9.2.5 nASAsaeettlotet2zedroeereCfrndoasa.ttltoalsente6avdxnoatCdrlruaad,crmsudtpeeedxsstotocarlaanutwcldtiitboeocrdnoaw.nt(eicroeeenkansgst.retaantntidobnalrsadnoskfi)nstma1nu5ds-mat rbLdespmporlaeyypparbroeepdpyrlweepnitaehreteudabceahss 10.0 Batch Set Up 10.1 10.2 ocpRiEnornaeonetcqhcthurreeoeidrbqaluesagumrteaacfnlelhointrtryetoctsciofafofinsesovstadrreumorrfylaipaen(tlflcmeodesrkeapnttehnhloxaedowtnrdbalnaafcbotbtclceroahodtrn.hankit(csoteuyrpnsypritionrcdajgauetlicplSoytln.imcs2aL0te(lsooarbfanmledcesotssnh)ptairmnkooelusls)tswapiniinlkdlcelb)utwedteosopareetvcaeilgferieaielnsydtt 11.0 Sample Extraction 11.1 Weigh 5 g of sample into 50 mL polypropylene centrifuge tubes (fortify as 11.2 nAededde5dm, rLepolafcmeettihdaannodl amnidxswhealkl)e.on a wrist action shaker for -15 minutes. 1111..34 TBrrainngsfethr ethevotulubmesetouapntuoltr4a0somniLc bwatihthanwdastoenricinatethfeor5-015mmLinpuotelys.propylene 11.5 cCeennttrriiffuuggeetfuober-.10 minutes at ~3000rpm. 11.6 Cfoollnodwiteiodnbyth5emCLnoSfPHEPLcaCrtwridagteers(~(12gd,ro6p/mseLc)).bDy opnasostinlegt c1o0lummLn rmunetdhraynol 11.7 11.8 LeE1l5oluuaamtdteeL.(wdpieothclyapn-5rt)opmtyhLleens1ea0mc0e%pnltermifoeuntgheathntueobl.ceosnC(dfoiintlliaoelcnvteod5luCmmnLe SPE cartridge. =of5 emluLa)t.e into Discard graduated 11.9 Analyze samples using electrospray LC/MS/MS. 12.0 Chromatography 12.1 12.2 SLaInntCjaae/nlcMydtzaSterh/ddMesssSaoamfmsPpyeFlesOatsem.Amo.uconArtroescfpaeolainbcdhraintsitgoanntodsaatrtadln,edasaasrtmdfpivlmeeuaosnrtdmpfoorerrectiecfdioeendcaesannmdtraptflieoolnlionlwteovteahllesl 12.3 bAminenutgesirtnesbpnnetieinrrisgenecdaslunebdtdeetowdafteineetnhaxentervaaeecnnrtaedyldy5ot-ic1fcaa0lalisbsasermaat.mtpiolpenlse.stsAaents.daanrEdaxslttermarncuatsettidvebs,etaannindecanlrutdidrseedmseuatstotfthbee Page5 of 7 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 27 o f65 Page 79 of 122 EOS-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: P0001131 Exygen Research Method Number V0001781 | ANALYTICAL METHOD Method of Analysis for the DeterminaLtiCon/MoSf P/MerSfluorooctanoic Acid (PFOA) in Soil by 12.4 12.5 Sgvse(cefUatooxaxeeomsllfcnrtleitrseoaebwpauecrwrlclsaadeaciterttnoeceesidrdeuoatdesaalnnsbyinrfctbpyossdaortftsrlaaeaoneitrttmnbarsxihdndoret.aeardanratesraasicdhssoenrtptdsocnaeaooludmnynnclsdtdsuuctdeaaeasrrnnsbvstldideoeybsbtatcsherlroooaiedtnnftufhxsioecloeecdraenexmfrnbetiqrtrrsaderueastyaatgacftsnnutaritbeeodntardeistnnadthsardsieuidltotanairsasornsjidndetnniiducn.glisrautnteeeirtjMnsddeerLpsgdcsapo)tiasi.anlentos/nerLsxnsdIateyenhrswidnrne.eexesaiaeitbtngaAvh3esanhe.eagnl3dtrmryiiyyannz(crpnogeods3ldriase-on.em1ecgsf,q0euppeutroel.sxvieaavfteskrmasaaltcapehtrnalsaeeerettdeastt 13.0 Acceptance Criteria 13.1 13.2 13.3 13.4 13.5 13.6 eTRMbwEHi2asRdmteACo1nhtxpl0na3hreafeaohjnaueecrptu.et0necr4wnry9iisoetlrcgrro%rodhu1tk9onrateehmvcacodek3bn2ptvtoTcraeiddensetreoadseolerr.adeieorhit(rirsnaodbnmbstoeR,ewoitbt.tolmsrgoelo,uaautJtatanrnrhiainl.ofuttammndtse2iiikvs(mhfonosst03Ttesaaeaenenctb.P6slypomh9ondmmuer9Fstse8neoebcapbatuuOl3tsataeoell4neersms.me)nvteoAv1dtten.wsdafeula3xusuIfntlxaesnashhicfs)umocrtceahttpeoeIardatilnmoebfeislewtubekeeceducpenfudenvecodtsoolrrtaetdtsaeen2onupiadolltbrspt0(tnafniasaaeberRbfye%rdnskdoifnegersanar)deealatrneoktmtnsrxrethootPstmmdeiftaotppeoomrfF-schtbatailfenreoOoeekttutecaxarahhrrsestnAtclirnreteeraotxadesaeahdadfsdhbsallsaactacssysipjuoueolnbttatpcuslsoaoelugtsarlartitsaldtncSalkhsemfeltidlouO.tids-ttevciboeuetplsosainfbeiarnxrntalcrnAulaetsetmgtblssdaiiotirmlsoot/nirodlearuLgocennyttnbuceosomrhuean,tmeeutnraueretdomtrumahvtthdasbtiesftsloPeetale.eon3eiiiaytxstdennFttr6fnrazhbireeiOn9cddeetaxegeoohbcdeaAtxetapinteawy.c.rhsfntmedpneapreeedarawstlurisuaeranirtekcsebahintbit-tiefifbtorheeenlotrealen7aldxognodatiLt,0mtmimn,ulroOt-siwakcthomi1nmtsatcoaaQre3hisnstinuued,c0iita.pdsdolsleHum%aedt,ttanrrhhuIvr7pttetfeabbhdghhoe0lninneeeeseee.saft an4a%nalwytiitchainl raunnatnhaelnyttihcealserut nm.uIsft rbeeternetainoanlytizmede.drift exceeds this limit within Page 6 of 7 Northern Alabama Potable Water Systems, December 2005 Page 28 o f 65 Page 80 of 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: P0001131 Exygeo Research Method Number V0001781 ________________________ ANALYTICAL METHOD_________________________ 1 Method of Analysis for the DeterminaLtiCon/MoSf P/MerSfluorooctanoic Acid (PFOA) in Soil bv 14.0 Calculations 14.1 bUgaessneeedrthatoeendfopblelyoatwkhieanrMgeaea)qssuuasLtiiynongnxtthsooefctsawtlacanurdelaaptrerdotgchruearmavme: o(luinnteaorfrPeFgOreAssifoonunpdar(aimn entge/rLs), PFOA found (ng/L) = /Peak aresalo-pientercept) x DF DF " factor by which the final volume was diluted, if necessary. 14.2 Fthoerfsoalmlopwliensgfeoqrtuiafiteiodnwtoithcaklcnuolwatne tahmeopuenrctsenotfrePcFoOveAryp.rior to extraction, use Recovery (%) - [ total analyte found (ng/L) analyte found in control (ng/L)];; analyteadded (ng/L) 14.3 Ungs/egt(hpepbf)o.llowing equation to convert the amount of PFOA found in ng/L to PFOA found (ppb) " [PFOA foundsa(mngp/lLe)wxeivgohltu(m5ege)xtracted (0.Q4D1 14.4 bUasseedthoenfdorlylowweiingghte.quation to calculate the amount of PFOA found in pph PFOA found (ppb) dry weight * PFOA found (ppb) x [100% / total so!ids(%)] Pige 7 of 7 im Report #18 Northern Alabama Potable Water Systems, December 2005 Page 29 o f 65 Page 81 of 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: P0001131 ANALYTICAL METHOD Method Number: V0001782 Method of Analysis for the SDeedteimrmenintabtyioLnCo/fMPSe/rMfluSorooctanoic Acid (PFOA) in Analytical Testing Facility: SE30txa5yt8egeCRnoeRslleeeagsreeca,hrPcDAhri1v6e801 "vOApprovedBy: - -CJ lL___ PTaeuchl nCiocanlnLolelayder, LC-MS, Exyg!en Research _D_a_t_elO k -b /o 1/ / /Voihc/ne FPlr?aehsiedrMetynt, /Operations, Exygen Research Date Total Pages: 7 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 30 o f 65 Page 82 of 122 E0S-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: POOOl 131 Exygen Research Method Number VOOOI782 | a n a l y t ic a l m e t h o d ____________________ j Method ofAnalysis for the DeterminatioLnCo/fMPSer/fMluSorooctanoic Acid (PFOA) in Sediment bv 1.0 Scope TaScphicidsctmbroyemthHeotirdgichisDtPoeetberfceotoermrm(apLnlCocye/MedLSif/qoMuriStdh)eiCnishsoreoldamitmiaoetnongat.rnadphqyuanctoiutaptlieodn otof paerftlaunodroemoctaMnoaiscs 2.0 Safety 22..21 pCArolewncasauuyltstioothnbess.earpvperosapfreialtaebMorSatDoSrybperfaocrteicheas.ndling any chemical for proper safety 3.0 Sample Requirement 333...123 33..54 pSNAAtSeraalmtoolmmjlpsseeppaaceallmmtrsee.atspptc3ullo0eerslestlpg.oemrcrooetucfidsoettenssbsrteipenfsrrtgaohimcgioseerpdronlaueeutreegfeddohselrydwesmxhiflotolirrxuabslceedetdidosbibnpmee.efeconirafteilelsbodaewmiinenpgdletshsa.teomspaelmeqdupilfliionbrgreapxteltaranctotfioornro.tohmis 4.0 Reagents and Standards 4.1 Water -HPLC grade 4444....5342 APMAemcereftmthliuacoonnrAooiluco-imcdtHa-AnPcRoLeietcCaagAtgeecrn-aitddAge-.rCaSd.iSeg.mRae-aAglednritcGhrade 5.0 Instrument and Equipment - 5.1 55555555555...........15147386129120 S5DA2AADsAW1p1o05P2Liimunlasds5EmmvathptpcoeL-leeimooovyvLprgnlcssnastiiLhticaaplddcnscSeebbeuiieaLaesseplltuctlrpeeptDpqoeemtbooHeurpmsPPacfssdiiPoEolm(apaapia1ctrkLblbpeonamln0rlleteConcVeeta0acdaeepir-ppatfvnsrti1wocc.oaoorpic0alaanlwleeid60ylywptdt.h-sp0pakcemldrbrcii(maptoqo5lao.(Lepup0vtuMlayiy-aogtda1lhlrfa)fneei0osarndbntcr0seCbeoehupa1lStrILecdcet0oSpleeai,e-mvennnk1Ss1ocgtt.0aPrr0tilt0nriuitE0ofooftmuu-pgem2c0ggLgrea0.aeeer0)r0pati,t0ettnuthruuw0riijLobbde0(i)eencLcgt1.hssateCao..pgsndr/a..MidbcsplaqSepou/MasaoabnbSflteil)tep.aouttfiimopisnnp..jiencgtinugp 5t-o2002 Page 2 oi 7 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 31 o f 65 Page 83 of 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: POOOl 131 Exygen Research Method Number VOOOI7S2 I _______________________ ANALYTICAL METHOD__________________________ j Method of Analysis for the DeterminatioLnCo/fMPeSr/fMluSorooctanoic Acid (PFOA) in Sediment b'v 555...111354 CWVeornritsterti-xfaeucrgt.ieoncasphaabkleer.of spinning 50 mL polypropylene tubes at 3000 rpm. 6.0 Chromatographic System 6.1 6666....2354 TG(MMAPern/ooamNabbdlp:iiyillee8etenir2caPPt5athhPul0aarr5Coess-eeo:g0lr(5(u3aAB2mm01)):n3C:::0F) lu2MomepthMhaasnAeomlRmP o(KnieuymstoAnceetSacteieinntiWfica)t,e2r.1 mm x 50 mm, 5p Tim2208e120....(0.0050min) 6226/6.55555A %3733755555B FimloLw00000V.....33333mRainte) 666...876 QRInuujenacnTttiiiotmanteiVo: no-:lu2Pm3eeam:ki1nA5urtepeasL.-(ceaxntebrneailnsctraenadsaerddtocaalsibmrauticohnacsu5rv0ep. L). TophteimabizoevtehceoHndPiLtiConssyastreemin.tended as a guide and may be changed in order to 7.0 MS/MS System 7.1 PMFoOdAe:. Electrospray Negative MRM mode, monitoring 413 -> 369 m/z for TophteimabizoevethceoMndSitMioSnssyasreteimn.tended as a guide and may be changed in order to 8.0 Preparation of Solutions 8.1 Mobile Phase 8.1.1 a2mmmMoniaummmaocneituatme taoc1et0a0t0e minLwoaftweratiesr.prepared by adding 0.154 g of Page 3 of 7 Northern Alabama Potable Water Systems, December 2005 Page 32 o f65 Page84oft22 E05-0210 Interim Report *18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: P000113l Exygen Research Method Number V0001782 | ANALYTICAL METHOD | Method ofAnalysis for the DeterminatioLnCo/fMPSer/fMluSorooclanoic Acid (PFOA) in Sediment bv' 8.2 Extraction Solutions 8.2.1 11%000acmetLicoafcwidatienr.water is prepared by adding 10 mL of acetic acid to Alternate volumes may be prepared. 9.0 Standard Preparation 9.1 99S..t11a..n12darmiAPomdnrfLeSea1atpth0noo1aaacf2rpnlle5tgycoh/t/amlFeimcisoLnaL1trlo0taLicsf0iokDti1acpr2Pantsgi5todEfi/-iloammcubrnatdLoLitSoti(toosLnlcoenloDu.olrustPrifoteoEi-locnu1tnbt0eiodo0ttontflpoeaog.rff/pmiPnuFLarOlitoyvAfo) lPiausFnmpdOerAdeopilabfuryte1e0dwt0obewyi1gi0bhth0riinnmmggeLi1tnh0gwanmi1othg0l 9.1.3 9.1.4 mAmaaA0L1L1122..1oo550ffmmpptthgghLLee/bmLnL11LLDD.00PPffppooEEggrr/tt/bbmiimffooiiLtLtccttaallseesttoii..ooollunnuttissioooonlnluuttttooiiooaannffooiinnffPaaPllFFvvOOooAllAuummiisseeppoorreeffpp11aa00rr00eeddwwbbiittyyhhbbmmrriieenntthgghaaiinnnnoggoll 1in0 1in0 9.1.5 A100m.01L pogf/mthLe f0o.r1tifpicga/tmioLn ssoolluuttiioonnotfoPaFOfAinails vporelupmareedobfy1b0r0ingwiintgh 9.1.6 aTfmrphoepetmhrosattnxohoiecmlkdianaatentaeldy1of24fo5`prCtmriefpLiacanaLrdtaDitoaiPonreEns.osbtloaubtttilloeen.fsoarrae mtoabxeimstuomredpeinrioadreofrfig6emraotonrthast 9.2 Standard Calibration Solutions 9.2.1 9.2.2 LoTfChte/hMefoS0l./l1MowpSignc/gmaliiLbs rfaaotrtiytoipnfiiccsaatatlinoednxaasrmdoslpualteiro:enap.dredpitaiorendalincomnectehnatnroatliovnias dilution may be SooCflouFntoicore1tt1n1oin5200f0(ot00irncagatt/iimoonnL) V(om1t11l25uo000Lm) e Dil(um1t1111ot00000eLo00000d) to Con(ncFgei/nnmtarlLat)ion 1250..00.0 001...520 Page 4 of 7 E0S-0210 Interim Report *18 Northern Alabama Potable Water Systems, December 2005 Page 33 o f 65 Page 85 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: P0001131 Exygcn Research Method Number V0001782 ANALYTICAL METHOD Method ofAnalysis for the DeterminatioLnCo/fMPSer/fMluSorooctanoic Acid (PFOA) in Sediment by 9.2.3 9.2.4 nSAatetloe2terdereCndaa.tltolec6vaolCilbu,rmuapteiostonanssidtxacnmodnoacnrdethnssti.rnati1o2n5s-mofLstLaDndPaErdnsamrroawy -bmeopurtehpabroetdtleass 10.0 Batch Set Up 10.1 10.2 ociREnonaentecqhchueeiunbrqentaurmttaarcetlehiianottytonesdsaffs(soslacarumobrfanipcentlorlcedonesltarpeonlxaaldtnnsrdlapafciobktreotewdtr)haotti(oostyrvpuyperniordctijraufeeylpcaltylpt.iercd2oa0cteecsodoruanrntlaredlosslrsse)pcimkofevouserstrtiwyfiiinefloldcrlbutedahetespbaekatcnticlofehiwae.sdnt 11.0 Sample Extraction 11.1 11.2 1111..34 11.5 11.6 111111...789 11.10 11.11 11.12 11.13 eSEAACcmfADC1LCCWAnfeWfooloe5lPelooeeudnoediudlunlraeenElannlcndmaandadttosdoi-udaettltrdtdgh6werryecweLitnt32iihiifehd.azwt0tff(tt5ceu0esiirhpe,udueodoogt.d5timoergeghmrtnsnlmemehbibllceeeagdicyepLayanyLmetgottmtcp-lnhhuhtuoasel5t2or2hpetuetebeeecofo.)0ea0lfdtmecepetsfthnsCm.ou1tmums.yaiaolnhmnnb%biuLmlnuClLdeLedeewesosnaptssSataotoisloC1iepehnlnctaPaaef0fop,hageudtm-ctE0HHnnt2-e-i4ev1i%tmnSpo33nPcP0ol1octllePm00tLiL.o0raeratx8cmEt00rhitCCcLet.fotm5orwe00ierxucod0wintowgeLsdatrhsr,hapfeplraapgamactltmemmmnto)threnratee.Lauidseopfedresrybffl,etgw(((.SoopshecL1d~ve-hrroPsoaaioCam-(-lnnt22CE(gy1re22kf/odt,oMrdipedece00lingnlr.rrt6xaalaioo,Sotremmoonlppt6p/aCcmndnrMlviiy/n/tienonsemsdLoldflecSlduuea5gtoll)Lccnttu.seeaee))iheC)mmcwb.nd..ssatby..ruLeckDCDotiytehshepn3ooontoeSpealota5PtlfsnnreanhebssioEolcecmfesiuottulntatiituaLnsglocgtlttewaeoehg)aenett.tcmerre1ctaolcit0srOseuononiihtlobdlndldummauuaegftakomcamsceLLtettdeo.hninr(g.donmfemronrdrfaiDueueroti5dtttsntnri0ihtiuhhsoofaddaacya-nmnntrartk3eeyhyoroaeL0dddelslr Pag 5 of 1 Report #18 Northern Alabama Potable Water Systems, December 2005 Page 34 o f 65 Page 86 of 122 EOS-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: POOOl 131 Exygen Research Method Number VOOOJ782 | ANALYTICAL METHOD I Method of Analysis for the DeterminatioLnCo/fMPSer/iMluSotooctanoic Acid (PFOA) in Sediment by 12.0 Chromatography 12.1 12.2 12.3 12.4 12.5 UvmsgseSLabbssiSIAntonttxeneeeataCaasnjufajrcngtamenennetws/nseleiwcdMcuddytnpederettalsaazabaindnlaieSorerrtrtenetircndhddeesin/nddearMstdresisgsaeaslnee)isinyr.nsfmsvcoaSibaoanspdsmlerafntusmaretoaareIPetsdpydtystrtnanmyerhiFadlenssoesbomae5.dOeapdesntrte-feme.eiaaipA1tsmnsrsntrhli0eohshpdtenaaace.xeuoeoljonnrssdtneyunrceaAdrectltacatsurmdeeatn.ceaenorervdtspsvsbacddfeneepellyasdeoeryo,ocslhayaslttinfotocibci.cncndfaeuhr5aatoaealcixhll-Anilrdatseic1sebibgtrnsaeso0rabqrebmrteantnaertueaoedts.atdigpnagiafotasoanrluisminetreotnnattradiasntlplnnt,shtsesnaliesdeitaueednstotrataisarassnga.onnrtimndranddn(dldoiutt.igapSlioavsfuvrlmrtrideeMteaenadeas,nusLgoasdcapdssnaricmseotn1aandmsomtnr/enLuuxpdasudfeofsynrsoerosnwtrenrtsrteltecmt.xilebsioraefciftbeeuongdawi3oeerAsnahe.netsdt3ldhtncedyaiiaenbnteasyz(ranngeocbesdomfledtdrsyiufrocair.apendcsomfcctfqltteuniaapoeeuopdrllderlaviiinllisabbnveonepaksrrsdalwtseteaaeolraetttvtsliiahrntteaooeaehhoratslnndleae!settfl 13.0 Acceptance Criteria 13.1 13.2 13.3 13.4 13.5 ATwEHei2sodbsteaMRC1nhtaxpn0alhf3raeaonjhaemrecptu.e0enrc4wnyr9ioiltrcrgr%rodophu1kr9oateehcvmacledk3on2pvetTaroedndecstersdealearre.meoidohrirt(isradmbtbsone,Rweoitu.tlomorsgtelou,ua1tsaanarnhr.nlftotuitmadteis2nivkb(smhfosot30sTaasteeaennctb6.lyohpm9omumnoPer9stesn8eeobcabpFtuulsat5aetlot4leOermssa.emnv)e1ovtteni.sAdafeun3lauxusInflaxsenshis)ucfmoractcdhtpaeoIadratimlobtieefstweubakecedpucenfdunelcedotoorrleteadasdenou2pnvdolstpbt(t0anteiafabaherRbfeyrn%dlskionfeeesranadr)eeltankottmsexnrgtohstPommenifttproeopor-FcfhtebaatieinflrooOekaretttecxaahrehrtestntlrAceireeeroaexadssaearafdsesdhallsaccatsspstyitjuouoltbtacthpulosemoalugtslarasittadlntcalkfhsletniuldo.uditstetcvbsiuoetslsoanfiebti0arrnatcrAnlauetbts.mlsbdsai2oitlimeonotiroldeurgocenynttnuerbonsohrauenemueetgnrureadmto-rmahtv/etbastiesmftlPoaxeetleni3eoiiaytxsdetFtnLr6trnrhafzbireieOa9,dcdetexegeochbdceaAtxtettaipawe.y.rchehsfntmdnepapdreeeaerasultrin.suneaaritkecsebahntibt-etiiofhrtaeeenloreane7ladxogonditL,nt0mtimu,lroe-Ostiwatc1imhmnwtsatocaaQhe3isntuiudn,i0cit.pdsllbodesHu%edat,ltnarrahuI7rvtttefngebbhhdho0enikneeeeeseasft Page 6 of 7 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 35 o f 65 Page 87 of 122 E05-0210 Interim Report HI8 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: P0001131 Exygen Research Method Number VOOO1782 | __________________ANALYTICAL METHOD_____________________ | Method of Analysis for the DeterminatioLnCo/fMPSer/fMluSorooctanoic Acid (PFOA) in Sediment bv' 13.6 Rane4tae%nnatliwyotinitchatiinlmrauensnatbnheaetlnwytteihecenalsersuttanmn.duIasfrtdrbesetearnentadinoanslaytmizmepdel.edsrimft uesxtceneodts dthriifst limmoitrewitthhainn 14.0 Calculations 14.1 gbUeasnseeedtrhaetoenfdopblleyoawtkhienagrMeeaaq)susuaLstiiynonngxttoshoecfatswltcaaunrldeaatperrdtohgceruaarmmve:ou(lnint eoafrPrFeOgrAesfsoiounndp(airnamnge/tmerLs!. PFOA found (ng/mL) = (Peak arselaop-eintercept! x DF DF - factor by which the final volume was diluted, if necessary. 14.2 Fthoerfsoalmlopwliensgfeoqrtuiafiteiodnwtoithcaklcnuolwatne tahme opuenrctsenotfrePcFoOveAryp.rior to extraction, use Recovery (%) - [total analyte found (ng/mL) - analyte found in control (ng/mL)] qq analyteadded (ng/mL) 14.3 nUgs/egt(hpepbfo).llowing equation to convert the amount of PFOA found in ng/mL to PFOA found (ppb) - (PFOA fousnadm(pnlge/mweLilgxhtfi(n5agl)volume (5 mLll 14.4 Ufosuendthien fpoplblobwaisnegd eoqnudartiyowne(iigfhnt.ecessary) to calculate the amount of PFOA PFOA found (ppb) dry weight - PFOA found (ppb) x [100% / total solids(%)| Pige 7 of 7 Report #18 Northern Alabama Potable Water Systems, December 2005 Page 36 o f 65 Page 88 of 122 EOS-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 200S Exygen Protocol Number: P0001131 ANALYTICAL METHOD Method Number: V0001783 Method of Analysis for the DeatnedrmCilnaamtisobnyoLf PCe/MrflSu/oMroSoctanolc Acid (PFOA) in Fish Analytical Testing Facility: SE30txa5yt8egeCRnoeRslleeeasgreeca,hrPcDAhri1v6e801 Approved By: TPaeuchl nCiocnalnLolelayder, LC-MS, Exygen Research DaItebh.( . M 'VJoichAen PFrlaehsiedretynt, Operations, Exygen Research Date Total Pages: 8 E0S-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 200S Page 37 o f65 Page 89 of 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: P0001131 Exygcn Research Method Number VOOOI783 _________________________ANALYTICAL m e t h o d _________________________ Method of Analysis for the DetermCilnaamtisobnyoLf POeMrflSu/oMroSoctanoic Add (PFOA) in Fish and 1.0 Scope TaScphieidsetmrboyemthHeotirdgichisDtPoeetebrfceotoremrm(apLnlCcoey/MedLSif/qoMuriStdh)eiCnihsforioslhamtiaaontnodgacrnlaadpmhqsyu.ancotiutaptlieodn otof paerftlaunodroemoctaMnaosics 2.0 Safety 22..12 ApCrolewncsaauuyltstioothnbses.earpvperosapfreialtaebMorSatDoSrybperfaocrteicheas.ndling any chemical for proper safety 3.0 Sample Requirement 33..12 3.3 pAfasSSaounrnaatoobdammljdlleeiyppmaclpsllestieer.aatsostv.2ciceoso0ehnlslgoo.seupooclertdfSinaotebennasidenltpphsrarafoonormmcodcezpeodepslguensleraefencdossietzrboweeertxahifwlotgelrireatebhcsetaoedimvoxsrnetpyprr.ealniecccisitegfi.iohienntdP. tlfiaornPcoleaztahlectlehneoewstshastaemofmrofparrplgoilnezecgesanuripbnnslotaaicnlnmontpdfitomliaeoriexntiienhdoriessaf 4.0 Reagents and Standards 4444444444444.............11817912531461320 TPSWAAMSFD1LC-euloi-amciOoleAramplrieftrutcbmcehtlsieeeaoutsrocaratoncinonohngln-lneirryeoe(ot(oibHr-lla6ul1loidin0(-ml2cP-cRie6-c0tLLa1H0eHah-A-cCC0-a4nlPPi2oHgc00od-L0LergeN0iPmo-t0CcnrCaHLamstemAtRidgCegjglseeacerherr--saniaaags)hddedgdhRrA-)eaeee)-R-e-.dnC-aeeSRtRg.RaigSeegegre.naamaeagtRgdgnaegeeee-ntnraAnattggtdglrgedeagrrnradraitadecdGdeheerade 5.0 Instrument and Equipment 5.1 55..23 AAAspoLnldvahceleiovygnnithtcinsceeaepctlqoetbeurcafdiooplatrlpolnmeecadcaetntwcarcaaneiwptdhaeldbmiaqaleutvMaiaodrfafiosarcrsebhpalSreedopaimevnkcogatiltrntuoootmgem0gre.aer0ptai0enthr0ijoe0(cnLc1atCaopgnr/a.MdcbaqlSepu/MaaobnSltfei)t.paotuifmoinnp.jiencgtinugp 5-to2002 Pag 2ofS E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 38 of 65 Page 90 of 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: POOOl I3l Exygea Research Method Number V0001783 _________________________ANALYTICAL METHOD_________________________ Method of Analysis for the DetermCilnaamtisobnyoLfCPe/MrflSu/oMroSoctanoic Acid (PFOA) in Fish and 5555555555555.............11111157918461356240 2AWDCT5DSR11105P2i2eumioisrs5snE5tmmtisppoLast-umorottpmrLLmiuycssafiLbpalaLucddieezbebetgiipLvaissestlleoearteeppDre(anp.coHo2pmsrPoasss-0iP(rpEapasihm1acLhabbeat0rnbaltolkCLote0eaplree)pe-r.peprvsr1i(..doopooiS0aelwlf0fulyytsl0s-kpppapmsierr(iptkoo5nl.oLcspp0nu.oyy-itac1nlhlneea0gdbnnt0.5eoeu1n0tLcc0toleem,-e.nn1sN1Ltt0.0rr00iip0ff5opuu-72lLggy10ee)p70,ttr7uuwuo)Lbb.pi)eety.hssl..edniseptousbaebsleatti2p0s0. 0 rpm. 6.0 Chromatographic System 6.1 A(Pn/Nal:y8ti2c5al05C-o0l5u2m1n3:0F) luopbase RP (Keystone Scientific), 2.1 mm x 50 mm, 5p 6666....5234 GTMMeroomabbdpiiilleeeenraPPtthPhuaarroessee:gr((3aABm0))":C:: 2MmetMhanAoml monium Acetate in Water lim22g08l20.t..Om..0050iB) %6662255555A 33377A55555S Flow Rate (mU00000.....m33333 in) 666...867 QRInuujneacnTttiiiotmanteiV:ono-:lu2mP3eeam:kin1A5urtepeasL.-(ceaxntebrneailnsctraenadseadrdtocaalisbmrauticohnacsu5rv0ep. L). TophteimabizoevethceoHndPiLtiConssyasrteemin.tended as a guide and may be changed in order to Page 3 of 8 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 39 o f 65 Page 91 of 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: P000II31 Exygen Research Method Number VOOO1783 ANALYTICAL METHOD_________________________ Method of Analysis for the DetermCilnaamtisobnyoLf CPe/MrflSu/oMroSoctanoic Acid (PFOA) in Fish and 7.0 MS/MS System 7.1 PMFoOdAe:. Electrospray Negative MRM mode, monitoring 413 -- 369 rtv/ lor oTphteimabizoevtehceoMndSiMtioSnssyasrteemin.tended as a guide and may be changed in order lo 8.0 Preparation of Solutions 8.1 Mobile Phase 8.1.1 2ammmMonaiummmaocneituamte taoce1t0at0e0imnLwoaftewraitserp.repared by adding 0.134 g of 8.2 Extraction Solutions 8.2.1 8.2.2 2o3ac%0fi%ddaimisDnceoi1tmrh0bye0iltcdhmiyacLclhdiodliocfirhmnolosmeirtloheastnahinelaaotnnlo.oelaiinfsintpoarlleuvpeoanrleeudmisbepyordefip1sa0sroemldvLinbwygi2bthrgitnooglufineagnsce3o. rmbilc Alternate volumes may be prepared. 9.0 Standard Preparation 9.1 Standard Stock/Fortification Solution 9.1.1 9.1.2 9.1.3 9.1.4 9.1.5 mPAAoiaaTmmmfA1nrpr0fheo1eLLep00atema1t2phr.mh.no.1o50osa1a0aattn1fxL2rfmnohplepot5iytochegmhlgpLtoaklme/idei/gmacmnfiLsaa/Ltn1a1mtetnLaDLlt.oe0l0haLdLycsP01feofDktp1of2Eao4fopgf2r5nP0rsoptrg/5bt.odiCEmtmrri1f/-oifletamimfiuitLbpcLrifacttcdjiaoLlLanaiiceastLogrttd(ai.ttoaiLsnrocDlitotloieoiaDoniunnooP.lonrntuLnPseEnifsetsooEo.-iossclbnos1lotutlobuonaue0tltlobttouidtu0titoitltolootteaeinflinopneo.oaffnog.srnioon/ffapfrmoiaPtrnuPlfaoeFLarFvPilOmttOoaooFyvAlaOfA)oubxfPmleiaAiiunmsFinsemsadOptiulospoermAdrerfvpeeoipolr1dpfpbauel0areyutpi10eernrma0ediwwrotd0aeoeebdidrwbityegho1oyif0bhbftrfm0hybiringi6re1nmbgmiet0nghmrr0Legaiai1ntnontih0owgngwonariglmtniinhIttoaihhggdsnIlt Page 4 ol'8 Report #18 Northern Alabama Potable Water Systems, December 2008 Page 40 o f65 Page 92 of 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: POOOl I3l LEx_My_gee_nt_hRo_eds_eo_arf_cAh_n_a_l_ys_i_s _fo_r_th_e_D_eatenrmCailnlaamytistobnicyoaLfCPle/MmrflSeu/otMrhoSoocdta_no_i_c_AM_c_eitd_ho_(dP_NF_Ou_mA_b_)er_inV_0F_0i_0s1h_7_8a3n_d 9.2 Standard Calibration Solutions 9.2.1 9.2.2 TopLrfhCetep/hMaefroSelldl/.oM0awpsSignnc/gemaeliLidsberfdaao.trtiytoipnfiiccsaatatlinoednxaasrmodlspulateiro:enap.dredpitaiorendalincomnectehnatnraotliovnias dilution may be S99oo..C22fluo..F34tnioco000ren..0t111.00in0anA.S...f120(000ti5teutrc55loea2tfaettdrt/ieirmeoCondnanLa.tlt)olec6vaolVCilb(uo,m52r1ml1111uau...0000L500ptmei)soteonanssidtxacnmodDnoacnri(dletum1111111hnst0000000seLt000i0000.rdn)atto1io2n5s-mo fLsCtLaoDn(n00ndc00oPF..00*e00..a.F0i0.o/.n000nrm0.02otd205ar155niLls55aat)mirornoayw-bme opurtehpabroetdtleass 10.0 Batch Set Up 10.1 10.2 ocRiEnoneanetcqhchueeiunrbqentaurmttaracetelhiianottytonesdsfafs(soslaacurmborfanipcentlorlcedonesltarpeonlxaaldtnnsrdlpaafciobktrteoewdtr)haotti(osotyrpvuypreniordctjiraueleylcpaltyltp.iecrd2oa0cteecsodoruanrnltaerdlossrlsse)pcimkofevouserstrtwiyfiiinefllocdlrbutedhateespbaekatcnticlofehiwae.sdnt 11.0 Sample Extraction 11.1 1111111l....S342 11.6 ddfePP(ctSAPWhofiariaailotmdyellarehcclarbd.ncoikkeetngoeiwtr3i2fhnahtzytb00henyh,eydeadle5mamdsancbt2itLLunigcdohy0rbehn-eooedeol1mmdfodsrffiayrgeeLt1aioicidtnfo2cnhseLr,SSenigotaalrCtoPnzaoetofnEh-enonprmrNineeellitwatretLHSru(iiczsilntibPeljeeaht.ehermEtrpal.oseifaendpCltoeuaDudilsrnoareebtti--sirnnnessm1eshddicesheaanaieaqmmhkatstrspiouonoed)oiee.lduoxnoudanSrfsnt.wlOcitDnilohelaeiwfanertmllynrwwl.aao)cisL.sizrgtoRkthiheheslseieupnttns2hmo.a.sfbelcelnygatypDistpsohefkrownleaosrorpinisfr-ntcylioshhsasohltaiisemnal2kkan,pg0eplew2lerlodemficwwgttoeefhLlinrlstatyt-htihsoorlIekiilfbScfusecua.mtefohmgnoegleeeuirtenehmlouta,3untunn0e2bcsos%eeetl.gos,,, Page 5 at'* im Report #18 Northern Alabama Potable Water Systems, December 2005 Page 4/ o f65 Page 93 of 122 Northern Alabama Potable Wat . . Exygen Protocol Number: POOO1131 Exygen Research Method Number VOOO178? ANALYTICAL METHOD Method of Analysis for the DetermCilnaamtisobnyoLf CPe/MrflSu/oMroSoctanoic Acid (PFOA) in Fish and 11.7 11.8 11.9 111111...111120 11.13 11.14 11.15 11.16 1111..1187 TeMPtetCDACASPtHfstAffDhhhholavxoahrleaeenddoeeeeemaaraatsrsanncradcmdkcnp--psktskapattelaiee11saorr.eycesoen2nii3fmar10stzt0fafgt0ettitahp-uru0euhrohere4-ttmeemt-mgmgenenhesshtsapmeehht.aiieiheiraLdsufznenmaez-aitaLrttbeeuausnpeehhpomoerxhrpettarxeteeeeepx-efstalholpdtwsdrdsesstrpaehai5lrSf.v.usaneicfaecOaf0otcclfoeucdalhtparaaeltatsctfousmmeogodss-iekofzdmknnan1frttLL1pnel.ieoogia0fean-rntttennloreoo,sppeHazCimdfikcsflsoolbteoaastePetorkllLasryrhtuicyyL.lwtlcnaelpr-ftheppoaeoCnoroe1hirorrcdrnldsfro0ooatulstadvdao-suppspttmiiictmeooicgnayyrthen1itaolehllgueiohtmsgeeynhmotsnennthSe2uuoniiahLebxeenlePstuieudsmCrei/tgiEronadrttansSeuu/iSLmiemttagMlxoseaPbbPecstrptnodeeEooErSiowyiilrssiasnnlf/elaseusccMivetcp.cct2uomotwtveohoooih%mSl.naltnrnsuneiuh.pina.ittmtsmezoaatbah1thei-risn2ltetnnnaaCeeruchSitciaobefofonpnteno$rmeiilirggtblodlr.p0tpue(Liene-sseaceccd3taamatntosssammr0.ih<itmLt-nlcahsappiss4bkhfdnelleir0cieeedaicnoezrpeieo.enmCnaealenutttdtd)thrmda.h-i-etpsft2e2eehfueta0m0leeghaani00lrasenoud-k00ntuisaonotrhttuhriirelaatnppbtinphstoammieodneeel.f 12.0 Chromatography 12.1 12.2 12.3 AmteaiiISLshnnnaintCntujtejmaaheees/nlecreecMpytdsntnrtlzbpeaetSdtedieecrshr/ddra.eMoeisssnseafesscaSodtAe,aalmftumescssPtpdovhaayeFlefaelmeesdinOacrtsbeypma.irAmbnllaa5oeielbt.cta-uigesr1oonnaierA0nrntnttari.naeosnosiatscanfntiSmpaalvegytoslneaapittnd,bcainlcodearhndaaasrifdannldtsra(igtssdotraaeeodnsntntmos.datmsiucsamaertcursettadutosln,steusdbftasenboaeasbttelrtmledfootfiohpinwifnrvelmteeececadlraufauossilsnrpidretbsbdecemtrryodpasfonartoeaeinrdrdoctecdtanitsebcfhlediieoletebsaetntdwrosbacatfenaseetignadsneiontmidnatannrjrneapndedctflisvieosntaoeitlrgoanmlirdonnnyasawltd)eony5.aivdn-rtea1hdbIall0neessatl 12.4 vUsgaeesmnresepurllsaietnceseadealtir.fborsrattathinoednaansrdtaalnycdtuearrvbdeyscloinnfoecraernqrtureagatnrieotisntsaiutoisoninnu.gsiMnLgaisn1s/eLxayrwnexsitga3nh.3dtian(rgodroecfqupureviavekaslaeranertea) software system. Page 6 of 8 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 42 o f 65 Page 94 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: POOOl 13 1 Exygen Research Method Number V0001783 ANALYTICAL m e t h o d Method of Analysis for the DetermCilnamatisobnyoLfCPe/MrflSu/oMroSoctanoic Acid (PFOA) in Fish and 12.5 eSxacmepedlesrtaenspdoarndseresshpoounlsdesnsohtouexldcebeedfusrtathnedradrdilurteesdpoannsderse.anAalnyzyedsa.mples that 13.0 Acceptance Criteria 13.1 13.2 13.3 13.4 13.5 TtmCodbmeEHMRAhnlahfrheonueuaertuec4rwiinyssotgroroh1ttkreehmcvcok3nbvtosTcaednoeeaeeolarroetrtirrtiombnobswe,eeioblttgorslxua,atnaatfrnhca.niotatmsneiiievkm(afnooseT3asamenncdtl6Pyhomdoump9eF2stneclaabutu0Osetaao4lnesr.s%smne1oAdttndfse3lusIfutnhoaxhhifa)somocracfptoedartimulteiwbeluheklcecpneudfdeneovorotraetsebnetnpuidoosprlte(tanasaeterRefrndaoirfnegasnl)derlctnkerot-snaeostetPmmfluotpaxomiFchmbbtaiftroeOekrturteaarbrrhaesAclriceteaoxttdeaithfrosseablacaasssipondulebtntaplo.oafgsrlrilntscsakh0efeiottdut-s.etcvaai5ueetslsoaneniaxrtcrnldspdttmtsbaioiirapaoodloarurcgbnntecnddosuhr,tssteaueretdothvathdbtiitftlnPoeehe.3eeijsxFenta6reahbirtO9ccdteaegthbcceAteeaawenyo.cdnmpneau.eeatwululnearitdsbahnbiifobtrenlrebaeln7ogdateL,0minloOi-twkemmh1nxaQhe3siucitc0p.asllsHu%eumat,rrduItvetpfbehghoneldeeeeeaft 13.6 2aRstp0eap.nt9erd9ona2ptrirdoi(sanRtoe!trimts2ht0eee.sp9rs8ebl5eem)tv.wuasentetInfsbeectsatoaltiafnbksdraeaamnrtdiopstnloeasrnaesddhsjuousluastlsmtdpfibnaleelslstrreuoammunutaesslniytdtzeneoodtpt.heedrsareitfiotlnimm, ioatrsne,dtthhteahnne an4a%nalwytiitchailnraunnathnaelnyttihcealserut nm.uIsft rbeeternetainoanlytizmede.drift exceeds this limit within 14.0 Calculations 14.1 UbgeasnseeedtrhaetoenfdopblleyoawtkhienagrMeeaaq)susuaLstiiyonnngxttoshoecfatswltcauanrldaetaperrdtohgceruaarmmve:ou(lnitneoafrPrFeOgrAesfsoiounndp(airnamnge/tneirLs), PFOA found (ng/mL) = (Peak aresalo-pientercept) 14.2 Use the following equation to convert the amount of PFOA found in ng/mL to ng/g (ppb). PFOA found (ppb) fPPOA founds(anmg/pmleLw) xeigfihnta(lgv)olume (m P x DF1 DF " factor by which the final volume was diluted, if necessary. Page 7 of 8 E05-0210 Interim Report 918 Northern Alabama Potable Water Systems, December 2005 Page 43 o f 65 ....Page95oTT22 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: POOOl I3l Exygen Research Method Number VOOOI78> | ANALYTIC.VL METHOD [ Method of Analysis for the DetermCilnaamtisobnyoLf CPe/MrflSu/oMroSoctanoic Acid (PFOA) in Fish and 14.3 Fthoerfsoalmlopwliensgfeoqrtuiafiteiodnwtoithcaklcnuolwatne tahmeopuenrctsenotfrePcFoOveAryp.rior to extraction, use Recovery(%) " [ total analytefound (ng/g) analyte found in control (ng/g)] ^ analyte added (ng/g) Page 8 of 8 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December2005 Page 44 o f65 .... Page 96 o1122 05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: P000113l AMNeAthLodYNTuImCbAeLr: MV0E0T01H7O84D Method of Analysis for theVDeegteetramtioinnatbiyonLoCf/MPeSr/fMluSorooctanoic Acid (PFOA) in Analytical Testing Facility: 3SE0txa5yt8egeCRnoeRslleeeagsreeca,hrPcDAhri1v6e801 Approved By: Technical Leader, LC-MS, Exygen Research Date Vloihcne PFrlaehsiedretynt, Operations, Exygen Research Total Pages: 7 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 45 o f 65 Page 97 of 122 EOS-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: POOOl 131 Exygen Research Method Number VOOOPS4 _M__e_th_o_d__o_f_A_n_a_l_y_s_is__fo_r__th_e__D_c_teAnNniAnLatbYiyoTnLIoCCf/AMPLeSrfM/lMuEoSrToHoeOtaDnoic Acid(PFOA) in Vegetation 1.0 Scope TaScphieidsctmrboyemtheHotridgichisDtPeoetebrcfeotoermrm(apLnlCocye/MedLSif/qoMuriStdh)eiCnihsvoreolgametitaoatntoioganrna.dphqyuancotiutaptlieodn otof paerftlaunodroemoetaMnoaiscs 2.0 Safety 22..21 ApClrowencaasyuuslttiootbhnsese.arvpeprsoapferialateboMraStoDrSy pbreafcotriecehsa.ndling any chemical for proper safety 3.0 Sample Requirement 33..12 3.3 apasSAfSounnraatoobadmmjlldleeyippmcalspelltsieer.asatos.tvc2iceoso0ehnlslgoo.seupocolertdfiSnaoteebnnaisdenltpphsraraoofnormcmodcezpeodepslguensleraefencdossietrzboweeertxahifwlotgelrireatbehcseatoedimvxosrntepyprr.ealniecccisitegfi.iohienntdP. tlfoiarnPcoletazahlectlehneoewstshastmaeofmropfarrplgoilnezecgesanuripbnnslotaaicnlnmontpdfitolmiaeoriexnitinhedoriesasf 4.0 Reagents and Standards 4.1 Water - HPLC grade 4444....5234 SACMiaclerietcbthaooanngnieot(rl1li(2l-6e00H---4P2H0L00P0CLmmgCeresagshdhr)ae)-d-eRReeaaggeennttggrraaddee 44444444........111691781320 PTASFLD1e-ulo-imoOArmplfrumceliseeustrctaonicoholnlnerrye(ooib6l-aulodin0mc-eiR-ctLa1eaHhAc0CanliPoc0go-dLerNeimo-tcnCaHsteAtRie2slggaechrr--naiaa)degdRdA-eee-R-e.nCaeSRtg.aiSgeeggran.maegtRdnaegee-ntraAatggdlgredearnraditdecGehrade 5.0 Instrument and Equipment 5.1 55..32 AAAspoLnldavhceleiovygnnithtcinsceeaeptclqoetbeurcafdioolpatlrpolnmeecacdaetntwcracaaneiwptdhaeldbimaqaleutvMaioadfrafoisarcresbhpaSlrdeeopaimvenkcogatiltnrtuoootmgem0gre.aer0pati0enth0irjoe0(ncLc1atCaopgn/ra.MdbclqaSepu/MaaobnSfltie)t.paotuifmoinnp.jiencgtinugp 5t-o2002 Page 2 of 7 Report #18 Northern Alabama Potable Water Systems, December 2005 Page 46 o f65 Page 98 of 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December2005 Exygen Protocol Number: POOOI131 Exygen Research Method Number V0001784 __________________________ANALYTICAL METHOD__________________________ Method of Analysis for the DeterminatbioynLoCf/MPeSrf/lMuoSrooctanoic Acid (PFOA) in Vegetation 55555.....58476 RD511025oi5stmmpamorLLysLaddebpiivssleaeppapoomross-rsaaiachbbtraolloeeprp.eppidpooellfyytlspapsrr(koSospOp.yy-lllOeennOeeuccLee,nn1ttr0rii0ffuu-2gg0ee0ttuuuLbb)ee.ss.. 5555555.......1111119135240 2SADCW1P2eumirsEn5tipoLst-rottpmiucsafibplLauceebegtLaistleorteDe(ncH2psPas0iP(Epphm1Laea0nbtLkCt0alee)er-vsrr1(..oioS0awfu0lsp0-kpmeipitln.ocLnuoitanchnagdbt.5o1n0tt0olm-e.1sNL0. 00pSop7lLy1)p7.7rwo).pityhledniseptousbaebsleatti2p0s0. 0 rpm. 6.0 Chromatographic System 6.1 66..32 MMGA(TPernoo/amNabbdlpii:yilleeeet8nir2caPPta5thhPul0aarrC5oses-eeog:0lr((5ua3AB2mm01)):n3C:::0F) lM2uomeptMhhaanAsoemlRmP o(KnieuymstoAnceetSacteieinntiWfica)t,e2r.1 mm x S0mm,5p Tim220e8I02...O..(0050tptn) %6266255555A A33755M5 I3S5 FfmloLw00000/.....3m3333Rianr)e 666...678 RIQnuujenacnTttiiiotmanteiV:ono-:lu2Pm3eeam:kin1A5urtepeasL.-(ceaxntebrneailnsctraenadsearddtocaalsibmrauticohnacsu5rv0ep. L). TophteimabizoevethceoHndPiLtiConssyastreemin.tended as a guide and may be changed in order to 7.0 MS/MS System 7.1 PMFoOdAe:. Electrospray Negative MRM mode, monitoring 413 -- 369 m/z for Pige 3of 7 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 47 o f65 ....Page 99of122 EOS-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: POOOl 131 Exygen Research Method Number V000J784 __________________________ANALYTICAL METHOD ~1 Method of Analysis for the DeterminatbiyonLCof/MPeSr/fMluoSrooctanoic Acid (PFOA) in Vegetation TophteimabizoevethceoMndSitMioSnssyasrteemin.tended as a guide and may be changed in order to 8.0 Preparation of Solutions 8.1 Mobile Phase 8.1.1 2ammmMonaiummmaocneituamte taoce1t0a0te0 imnLwoaftewraitserp.repared by adding 0.154 g of 8.2 Extraction Solutions 8.2.1 8.2.2 oa32c0%fi%ddiamisDnceoi1tmrh0bye0iltchdmiyacLclhdiodliocfirhnmolosmeirtlohaesnathinelaaotnnlo.oealiinfsintpoarlleuvpeoanrleuedmisbepyordfeip1sa0sroemldvLibnwygib2thrgitnooglfuineagnsce3o. rmbiLc Alternate volumes may be prepared. 9.0 Standard Preparation 9.1 Standard Stock/Fortification Solution 9.1.1 9.1.2 9.1.3 9.1.4 9.1.5 mfTaAAAommimaP1rnp0hrofe10eLLpea0et1mat2.hprm.h1.ono50soa10aaattfn1Lf2xrhonmptlpeo5ityhecogmgphlLtaoklem/dii/egmacnmfiLas/aLnt1am1ttenLatDLl.oe0h0ladLLcysP10eoffDkpt1of2Eoa4fopfg2SrP0nrosptrg5/bt.oiCdEtmrmri1f-/iotfelammfiiiutbLcpLirfactctdpioaLLlanaiceasLtgorttdai.(totaiLs/noDlicotmtleooianDoiuno.PoonlrrLtuPnseEnfirsstooe~Eo.isoslnbcsol1toulotuobanuet0ltltbotouitduito0tiltoltoottaeeniifnlopn.oeoaffongs.noironf/fafpromPairtnuPfloaFeLaPrFvOlimtFtoOoaoyvAlOaAfu)boxfmeAPliaiiunsmiFnsesamdpOtiluosporemAderrvfpeeopiorld1papfbuel0arueypti10erenrmae0diwwrodta0eeobedidrbwytieohg1yoifbbh0ftrfmyhb0riini6g1renmgbimet0nghmrr0eLgaiia1nntonith0nwoggwonaigrlmtniinh1ttaoihghgn0sItl 9.2 Standard Calibration Solutions 9.2.1 LoCft/hMeS1/.M0 pSgc/maliLbrfaotriotinficsatatinodnasrdosluatiroenp.repared in methanol via dilution Pu*< 4 111 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 48 o f 65 Page 100 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 200S Exygen Protocol Number: POOOl 131 E_x_yg_e_n_R_es_ea_r_ch____________ ANALYTICAL Method Number VOOO1784 method__________________ | Method of Analysis for the DeterminabtiyonLoCf/MPeSr/fMluoSrooctanoic Acid (PFOA) in Vegetation 9.2.2 pTrheepafroeldloawsinnegeidseda.typical example: additional concentrations may be SooClfuoFtnoicorentinf(tiirciafattti/iomonnL) ll..OO 9.2.300o0..100..oS.1200st55ore all V(om251l.u..L500m) e calibr1111a0000tion standDari(dlum1111111s0t000000Le0i000000dn) to125-mLCLoD(n0U0c000PF..00Re0.0..i00E.0.nn000/0002mta2105r15n5l5a5Lat)irornow-mouth bottles 9.2.4 Aatl2terCnattoe6voClu,mupestoansidxcmonocnethnst.rations of standards may be prepared as needed. 10.0 Batch Set Up 10.1 10.2 ocEiRnonaenetcqhchueeiunbrqnetaurmttaracetlehiianottyotnessdaffs(soslaacurmobrfanipcentlorlcedonesltarpeonlxaaldtnnrsdalpafciobktrteoewdtr)haot(itosotyrpvupyreniorctdijraufeeylcpaltyltp.iercd2oa0cteecosdoruanrntlaerdolsslrsse)pcimokfveouserstrtiwyfiiineflodlcrlbutedhateespbakeatcnticlofehwiaesndt 11.0 Sample Extraction 11.1 1111..23 1111..54 ctPP(fWAChfoaaaoederreccnbrnd-ikkttgo1ir32fhanit0y0f0hn,uedma5mamgsnce2iLLgnndo0teunohoetoImdfeedffisagLeta5.icdfco0eLr,SenotCtroPmzoten-EhenpLNineilttatrHSpruicsilobPjleea.elemEyal.sinpdCptDudirlnooaebisnnpsehsddciyseanailqmaktteriounednoidexenoanSsnlwOcttliuheleawabemlnwewla)cisL.sizrothiechlseupottsmh2ona.eltcnyagtDpsiipnoefrowinlaonorpnisgr-tyioshhshlstaieala2kan,mp0elele2rpodmflwcgeoefLlnrstaa-htiostrlIekiifS-cfs2cua.mog0megle0uitenh0mltu.aunntr2beposemtl.ogs, 11.6 ddSforiimyllal.oenwtihzeyedldbitchyhelmore1ot2hs5ialnaonmleLi(nthtrpoeeleuaert-insmeheassop)le.udtiDonfrly.asRtkhsiensfeblaytshkesrfinlcasosimkngpwlietwtheilttyohlubeetnfhoeereon3uc0se%e,. 11.7 tefDhliaeteshckpea.renabtryt-hsaehiare-pxdetrdryaicfnltgaosoknr. twoCiatohlclaeocnsttdrtiethaieomneeoludfanSteiPtrEiongcteohnleu. m12n5fmittLedsiinlasnidizeedthepemaro-ushthapoef Page 5ol`' Report #18 Northern Alabama Potable Water Systems, December 2005 Page 49 o f 65 Page 101 o f 122 EOS-0210 Interim Report #19 Northern Alabama Potable Water Systems, December 2005 Exygcn Protocol Number: POOOl 13 1 Exygen Research Method Number VOOt)I?S4 analytical m ethod Method of Analysis for the DeterminatbiyonLoCf/MPeSrf/MluoSrooctanoic Acid (PFOA) in Vegetation 111111...8910 11.11 1111..1132 11.14 1111..1156 estMATDfeACRSAhoavxhreenededmaraatapncadrdkp-npkaaetle5eoesyrc2enai3fraztt0ftteaimpt-reuhohsr4t-emtgeetniseshtaenheh.aLerapdfsuame-itatsrtsnpoeheomrehpaex1esfepxalld.p1dteastpr.l5ruvsca8feea0coelcoudcataettlsfthosumgdsikornmnafoLpt1niliagotaue-rntrnoeso,pgeHidfklcslohbeoestPtslayhurcytL1wnaeotr-p1anroCe1io.rdtr!ds1ouh0lsadv1seppttmiimoiocnryaasnaoelglagimgsyenmtmahnS2uuiaiLbxneepPstrmCie/.ilEondtnese/uLtieMgxasiobcnsrtndeooSroywstialf/lahsucMviepcte2mtwvehoo%S.a5nrsnip.i0na.ttsoaabhtmr-siletanCacehLtcioeoopnterclirigbllopepue(inenacesectatatntssrmar<.thi-tcfahspui4hkdhegl0eeaoerepirnC.altneuut)dmab.-tet2eehf.t0lehai0sank0ntionoirlatptinhatmdoel 12.0 Chromatography 12.1 12.2 12.3 12.4 12.5 USe(cgvseALambifSIntonoxxaneeaetoCsntuljaflcnrtamgeeleitrss/neloaebiwracMuytpneedsrcwrctlszabapndlnacaitSeetertneeettirocheeisi/dnddrdeuarMordetisgseasaenlnsnebiyfcnrssaScobtdoyapsasadmslrffsuttmrnleoaabeoatiPespdtdtntarbmneyrexhiFldensrotdestoead.waOeaadrsrtnaftaemet.aasrieiAcmsnsdsonereeohttdptcsnanhea.xacuoooldnemnotnyunenrcsAdrttlavatsuucdreedaacenoeesarnrntbsvssatrcldfneddiyepleyabsbdeoyatosclhearti5otlroonaobiicdct-ncffnetduhfhr1asioaeaxciloel0ealnradtsiecnmxibrtgsfneosbatsiqerreatasrnrntaaeteruadsotmy.mtadagtaciftasnoparuntaispbteerodtnltnrtaldeedasitnaeldnth,ssnerasd.iseuadisdltotstnaairasassaaenorjstmAindrtnednnt.idfduidcnsp.gliiastuavntlmeraierereMtjeEndddenerusLsgaoxsdcsapspan)ttairstoi1r.naeldtmsoamtnn/reeLxpncsIsdtfuarotheynroenrwseierdrensrntaedceete.xstieaiaeiicftbbtgvdnaisvoh3eeseAhetneaene.aagd,3ltdrnrcmnyiiiyannnaeasydz(crpannnngoceaad5mldlitsdrrseuno-redae.1apedngcfmss,t0qfleuetpieoieuoptdrorem.lxsvinleliafavetoneukramsaslwtsasateoeltcapevtttrhantltseeeahbehoaretldaleseee)sttlf 13.0 Acceptance Criteria 13.1 dCoafhu4rgo1hm3teaartmoiogunr.a(m3T6hm9eua4sm1t3ush)aormewpuraepspaeerneatnksttochfoearlrodessaspuoognhfdtcesarrtbiooontnhadetioP3xF6iO9deAa.mauniofrno,mwahiplearethnet Pigfi 6 of 7 Report #18 Northern Alabama Potable Water Systems, December 2005 Page SO o f 65 Page 102 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: P000I I3l Exygen Research Method Number V0001784 __________________________ANALYTICAL m e t h o d __________________________ Method of Analysis for the DeterminabtiyonLoCf/MPeSrf/MluoSrooctanoic Add (PFOA) in Vegetation 13.2 13.3 13.4 13.5 13.6 aTtHmasRRbmeEMAhtnp0nlhreeaon4ueaueptr.tcneaw9iniysosterr%ordnht9tokrneevaccaokn2bptvTleoarcsiwdnoeryeodelorrieoti(tiriarntsbonbsiteR,weetchoblttorsexla,aatJtiantafrhcinnailottmmsneitesiivkrfhaonaoste0uaseaemnnecendt.nlsyPpo9odumrpas2tFst8neecbanlhbatu0Olst5eoaleaneeerms.%s)netolvAndtn.wyfdusleaIufntothtaxsenaiifshhmoocctrfecpttIeeadtainultibfselhublkeslceceenfddeeenrceovsotrotuettatsentbnouaindoolmtlrtet(annifas.aetrRbfnadduskoirfgrsnldeIar)caeslaeroftm-tnranteosemPtdmriflbupaoexpiomFsebmatbtitntlreoOkeretrtueraabhrranesesAicnrcteetaxtaatesiatdfihrondhebosalacassoindaujolnebntaptusll.flauslayrritstslcts0kasliemtezitodmtuts.-eaveai5uelptsnifodbneeainxtalcd.nlepsdtemlstsdaiialrpasotdrrlarurrgcebinuedcdofsomuar.ttmsutesnurteotuaevhthabdtietsfstxlneelheen.ieitycsdjenatrteazhebterncceeteaegoohtcbcddteetptnaewoyc.hsdnenpeuae.eedtrwtullshaearitrdsebhainitbifisbtrenlotebael7llgndaietiL0mmimn,lo-Otiek1ohmimniaxttQr3esnscieuw,0tc.adlssHu%umit,tttrdhhuhIvpttefeahhgbioelndnneeeeea.f 14.0 Calculations 14.1 bgUeasnseeedtrhaetoenfdopblleyoawtkhienagrMeeaaq)susuaLstiiyonnngxttoshoecfatswltcauanrldeaatperrdtohgceruaarmmve:ou(lnint eoafrPrFeOgrAesfsoiounndp(airnamnge/tmerLs), PFOA found (ng/mL) - (Peak aresalo-pientercept) 14.2 Ungs/egt(hpepfbo)l.lowing equation to convert the amount of PFOA found in ng/mL to PFOA found (ppb) = IPFOA founds(anmg/pmleLw) exigfihnta(lgv)olume (mL) x DF1 DF " factor by which the final volume was diluted, if necessary. 14.3 Fthoerfsoalmlopwliensgfeoqrtuiafiteiodnwtoithcaklcnuolwatne tahmeopuenrctsenotfrePcFoOveAryp.rior to extraction, use Recovery (% )- [total analytefound(ng/g) - analyte found in control (ngig)} ^ analyte added (ng/g) Page 7 o i" im Report #18 Northern Alabama Potable Water Systems, December 2005 Page 51 o f 65 Page 103 o f 12T EOS-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number. POCO113 1 AMNeAthLodYNTuICmAbeLr MV0E0T01H7O85D Method of A iilyilt for the DMeatemrmmianlaLtiiovnerobfyPLerCfl/uMoSro/MocStanolc Acid (PFOA) in Small Analytical Testing Facility: SE30txaSyte8geCRnoeRlsleeeagsreeca,hrPcDAhri1v6e801 Approved By: tU f-lL TPaeuchl nCiocanlnLolelayder LC-MS, Exyg1en Research _ Date / JVgoihcne FPiglraehsiedre/tynt,. Oj pn/eratio-rns, Exygen Research Date Total Pages: 7 Page 52 o f 65 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 .........-- Page 104 o f 122 05-0210 Interim Report 918 Northern Albeme Poteble Water Systems, December 2005 Exygen Protocol Number: P0001131 Exygen Research Method Number V00G178$ | MethodofAnalysisfortheDMAeaAtmerAmmLaiYnlaLTtii1ovCneArobLfyPMLerCEil/TuMoHrSoO/oMDcStanoicAcid(PFOA) inSmall 1.0 Scope TaScphieidsctmbroyemthHeotdirgicihsDtPoeetberfceotoermrm(apLnlCocye/MedLSif/qoMuriStdh)eiCnihssormolamatlilaotmnogaamrnadmphqayulalnicvtoieutrap.tlieodn otof pearftlaunodroeomctaMnoaiscs 2.0 Safety 22..12 CAprolewncsaauuyltstioothnbess.earpvperosapfreialtaebMorSatDoSrybperfaocrteicheas.ndling any chemical for proper safely 3.0 Sample Requirement 33..12 aAsaSSfounnatgobdam)ldlle,yipmaltpselhsieraasteostv.n5iceosengAhnsooo.slotpuopeflerrrdtSnnoeaesacbnatetiedenlsslpsahyaimrof,nnormgopdicfzleoietesspghnslfneeeaonerdcrsecietzeeboiexsserttsahfwraaogeanrircetyethsiinaaooednmsnvxrud.teyprfrtaflnhieiccciestgeiie.oshinanntPmt. tlafpoarmPcloeleoazaceluctlanhenonetwtshbostaeeoffmrosufaarrpsgomelezcedpeasnlaureibsnnsot(sainc-ulmlopentppdsitlmlisaeioeiextndihine.adornesaf 3.3 pSraomjepclte. collection procedures will be specified in the sampling plan for this 4.0 Reagents and Standards 44444.....13542 PMAAWemcearetftmhtleouaronon-nirotioHrulioml-PcetLH-aACnPHcoLgeiPtCrcaLatAdCgeerc-agiddrAae-d.CeS.iSg.mRae-aAglednritcGhrade 5.0 Instrument and Equipment 5.1 5555555.......8523746 2AA5DsA1p1o50L2mnilds5vmmahpceL-leiovoymLLgncnisthtciLladdnsceebiieLaasespltclrpepDqoetbHooeurmPcafdssiPoolpEaaiatcrLlpbbonlmnrelelCoceeaadcaeprtppvntriwacroopoicaaanleweliwlyyptdths-kpapelmdibrirm(atqooa5l.oeuppt0vuMaiyy-aodt1llfhrafeeoi0sarbnncr0seboeehpualStredecLctoplaeeie,mevnknnsc1ogtt.atirr0ltrnituio0ofoftmuug-em20gggre0a.eeer0p0taie0ttntuhuuri0jLobbe0(nc)eeLc1.asstCao.p.gnr/a.MdbclaqSepu/MaaobnSfltei)t.paoutifmoinnpjiencgtinugp to 5-2iX 2 > Page 2o f R epartas Northern Alebeme Poteble Water Systems, December 2005 Page S3 o f 65 Pege 105 fT2T EOS-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: POOOl 13 1 Exygeti Research Method Number VOOOI785 | ANALYTICAL m e t h o d ~ Method ofAnalysis for the DMeatmermmainlaLtiiovnerobfyPLerCfl/uMorSo/MocStanoic Acid (PFOA) in Small 555...11910 AWDuiastptoeoprssipaSbeetltepepsPi(ap1ke0tVt0e-as1c.0600ccpL(lga)ndtC1l08-1S0P0EpcLa)r,trwidigthesd.isposable tips. 5.12 SPE vacuum manifold. 555...111534 CTWeisnrsitsurtie-famucgitezioecnra.pshaabkleero. f spinning 15 mL polypropylene tubes at 3000 ipm. 6.0 Chromatographic System 6.1 6666....5234 TGM(AMPerno/omaNabbdlp:iiyilleeee8trni2acPPtt5ahhPul0aarr5Ceoss-e:eog0lr((5u3aBA2mm0)1)n:3C:::0F) lM2uoempthMhaansAeolmRmP o(KnieuymstoAnceetSacteieinntiWfica)t.e2r.1 mm x 50 mm. 5p Tim2208e120....(.00050min) %6626255555A %3337755555B FfmloLw00000/.....m33333Riaritle 666...687 RIQnuujenacnTttiiiotmanteiVo: no~:lu2mP3eeam:kin1A5urtepeasL.-(ceaxntebrneailnsctraenadseadrdtocaalsibmrauticohnacsu5rv0ep. L). oTphteimabizoevtehceoHndPiLtiConssyastreemin. tended as a guide and may be changed in order to 7.0 MS/MS System 7.1 MPFoOdAe:. Electrospray Negative MRM mode, monitoring413 -* 369 m/z for TophteimaibzoevtehecMonSdMitiSonssysaterme .intended as a guide and may be changed in order to Page 3 of 7 Report #18 Northern Alabama Potable Water Systems, December 2005 Page 54 o f 65 Page 106 of 122 EOS-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: P0001131 Exygen Research Method Number VOOQ1785 | a N /vL V T IC A L M E T H O D I Method of Analysis for the DMeatemrmmainlaLtiiovnerobfyPeLrCfl/uMorSo/oMcStanoic Acid (PFOA) in Small 8.0 Preparation of Solutions 8.1 Mobile Phase 8.1.1 2ammmMonaiummmaocneituamte taoce1t0at0e0imn wl oatfewr aitserp.repared by adding 0.154 g of Alternate volumes may be prepared. 9.0 Standard Preparation 9.1 99S99....t1111a....n1324darAmaATommiafPdnrprfhoe1LSLpe0ae1mat2pt.rho.no15oos10aaafcttx2frmhnoptlkpe5iyhteocgm/ghLtFaeklm/ide/macmoiLsaatL1nar1tetLnDL.ltoe0l0aidLycfsP0fofiDptko1fcEoa4fogpr2aPnrsptr/g5tbtidoCEmtirif/-oifoealmimfiuLtcbprniactdctaLolLanaieSastorttdi(.ottaiLosonlciolteoolanDouinuo.olnrtnustePnifsiesootooE.i-osclnonl1utltubanue0tttbooidti0otitloooteanfnlineoiaffog.sorionf/ffparmaiPrnuPlaeFLarFvilOmttoOooyvAlaA)fuobxmPleaiiumsinFsesmdOptuoopremAderrfeeoipl1dppabuf0areyti10erenre0diwwodta0obedirbtwiyehg1oyfihb0trfmhb0riingi6ernmigmetnhgmrLeagai1ntonti1ho)nognwalrgtmnih1taio1snhgtIl) 9.2 Standard Calibration Solutions 9.2.1 9.2.2 pLoTrfChetep/hMaefroSe0ld/l.Mo1awpSsignnc/egmaeliidLsberfdaao.ttriytoipnfiiccsaatatlinoednxaarsmdoslpulateiro:enap.dredpitaiorendalincomnectehnatnraotliovnias dilution may he S99o.o.C22flou..F34ntoicoret1152tn1oin00ASa...f000(ot00ittrncloa2tfatetrti/eiromConnanLatlt)olec6vaolVCilb(uo,m25r1l11m1ua.u..000L000ptmei)soteonanssidtxacnmodDnoancrild(etum111111hsnt000000seLt000000i.rdn)attoi1o2n5s-mofLsCLtaoDnn(ncdPFge00520a1iE/n.n...r..m152000tdarnlLsaat)imroronawy -bmeopurethpabroetdtleass needed. Page aof ? E05-02101rrterlm Report #18 Northern Alabama Potable Water Systems, December 2005 Page 55 o f65 Page 107 o f 122 E0S-0210 interim Report *18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: P0001131 Exy_gMe_ne_Rt_heo_sed_aro_chf_A__na_l_ys_is__fo_r_th_e_DMAeatNemrAmmLainYlaLTtiiIovCneArobLfyPMeLrCfEl/uTMoHrSoO/oMDcSt_an_o_i_cM_A_ect_hido_d_(NP_uF_mO_bA_er)_Vi_n0_0S0_m1_7a8_l3l 10.0 Batch Set Up 10.1 10.2 oEciRnonaenectqhchueeiunbrqentaurmttaracetelhiianottytonesdsaffs(soslacarumobrfanipcentlorlecdonseltarpeonlxaaldtnnrsdalpafciobktetroewdtr)haot(itosotyrvpupyerniorcdtijraufeeylpcaltyltp.iecrd2oa0cteecsodoruanrntlaredlosslrss)epcimkofevousresttriwyfiiinefllocdlrbutedhaetespbaekatcnticlofehwiaesndt 11.0 Sample Extraction 11.1 111111...324 111111...675 TACDAWnmdHeireeoddesaeencpamdddniatsgoresw5ounhsifdfgrtaeam,ueebr1ttdgrnehlLegeerie1dpzotcteoleohseafmpufensctaepahseLtcntraemeueidlmrfbtisnpoiuodenapalfgnsemgalteieanattuopnrhttdisinulten-liebemn3ttioehfng0asio.eatxn0aaormds0wat5aapis0mrsefhlpsiel5namlpum0)akle.uLelemmfsvNsofipiiLonorzozolgre-etulde5ry-mai2apfstvmohe0rpaoradoioimpnt-lisafs1yuaaipbtl1lnbeetolm0lueesnsert.imaeenfncbosauLcelrttoee.neeunnt.srwtpeirafi.iepufwiugegrgtehitestestl-utauoibncbfetteiolosiavnn(eadfrsohm1raat5dikafdyeymr3.baL5es 11.8 1111..910 11.11 AmCLEf1o5olonLlualanmotdoledwyLiftzthwewipeodeioasntsbtlhaeyaymtrmph-.5rp2epolmlepCemsyLnoluLenosnSifconePHogfcEnPemedLnlcieettaCrictroihttfwrnaruoinegdasdotegpelerC.trausfuyb~(Ce1LS2oC(PlfgdlE/i,eMnrocca6tpSal /r/mv2sMtroeLiclmSdu)).g.mLeb.eDyo"oDfpnia2essolcmsutaiaLnlretdge)t.eci1lonu0ltauotmme.aLn rmunetdhraynol graduated 12.0 Chromatography 12.1 12.2 12.3 tALaSesmIiihnnnanitCnteujtmjaaheee/nsleceMrcepytdsntrtnlzbepaeStdtedierceshr/dd.arMoeeisssnsfaesesScaotdeA,aamlftumescssPtpodvhaayeFlefaemleesdiOnacrtsbeypma.iArmblnlaae5oielbt.cta-uigesr1oonniaerA0nnrtnttar.inaoesnositacsanfntiSmpavalgeytsloneaapitt,dnbnaiclcodaerhndaaarsifdannldtsrai(tgssodtraaeeodnsntntmos.datmsiuascmaerturcesttadutosln,tseusbdftasenboeaatsbtelrmtledofotfiohpiniwfvnrelmteeeeccadrlaufauosslispnirdretbsdbeecmtrryodpasfonatoreaienrddrotcecdtnaistebfchlediioeleetbsaetntdrowsbacatfneaseetigandnseionmtdiantannrjrenapndecftdlisvioetsnaoietlgornmalirdonnynasawltdeo)ny5.aivdn-rte1ahhdIall0eneasslt 12.4 sUasmeplleinseeatr. standard curves for quantitation. Linear standard curves are generated for the analyte by linear regression using 1/x weighting ofpeak area Pge5of' E05-0210 Interim Report *18 Northern Alabama Potable Water Systems, December 2005 Page 56 o f 65 Page 108 01122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: P0001131 Exygen Research Method Number V0001785 1 ANALYTICAL METHOD | Method of Analysis for the DMeatmermmainlaLtiiovnerobfyPeLrCfl/uMorSo/oMcStanoic Acid (PFOA) in Small 12.5 eSvsoxeafcrmtsewupesadlercesartsaeliynsbspdrtoaaerntmidso.enressshptaoonundlsdaersdnsochtooneuxcldecnebeterdaftuisortatnhneudrsaidrndiglurMteesdapsoasnnLsdyenrse.xan3Aa.3lnyyz(oedsra.emqupilvesaleihnati) 13.0 Acceptance Criteria 13.1 13.2 13.3 CobtRdmeM1hnla3fheaeuetu0cr4inistgor%orh1tkehmv3kobtcednseeasoareotrhiomnbotewoibgtlosouuaatnrna.inlofandivkm(fnss3Tasaecb6lPhmdomume9FenepauuOstae4nlsrs.mevo1dAttsal3suIntlhshasuf)oahpotearatmoiewtlkeecepcuunedovorlastdeennpisplbrttaasbeeareyrnoineeagsndltnkertsrhetsPtmeotpaem-FchtaiefoeOektuxaarrresnltrAiorxtdteahfassbalacaassypuelttnplsoegosirtnldhkfe1eod.-tect0veueossaerxtnrAlstmtdbsgioirondeo/auggnyttencsher,tteareetdmthmatbdhitoPeae.e3eitxFnn6rrabiiOe9cdxteahceAtaiewan.sfmpnepeatwruineatekhnib-eofbeelrne7lxooa,L0mtunlr-iOwatkm1sacQh3istidi0tipa.lsoe%ema,nrI7etptfhho0nlieeeastf 13.4 13.5 13.6 s1aHmART2awEtnp0ah4reoanurp.nterawy9soerrd%ntonr9aeacacnntp2vrTlaoiwotrdyoelereititisr(nradbsit,Reetch.otrxel,a3ttaiarcihnltmtmteise2irhoaoteu0eaeenndntns.ypo9ra2stset8bcabnht0l5aoelaeeem%n)telvnnw.ydfeuaufottxaesniihmccrftectIedainltsfbeuhblseenfdeeecrtsortuteatotuandolmot(nfina.tRbfadskdufrarl)Iescaaomtftnrnatomdriflubpoeiosmebbtltnroeetrerabhansecnreettaaaseaidirnodhoslcsoinaujonbtasulaflualyrtsttscsalsmiztidttmutseaaiptlfoidnbnieanalc.ndedtelsdailaasotrlrrrrecinuddomfoautmsusnurouetvatietsfsxlnlehniityjcdsetatezhetrenceeegotobdcdeetpto.ychdsneau.edetrlulsharirdsebaiitifsitrntoeballgdnteiimimm,oteonhmiiatxtresneucw,cdlsHuuittttrdhhuhvteaegbhiennndeee 14.0 Calculations 14.1 bgUeasnseeedtrhaeotenfdopblleyoawtkhienagrMeeaaq)susuasLtiiynongnxttohsoecfatslwtcauanrldaeatperrdtohgceruaarmmve:ou(lnint eoafrPrFeOgrAesfsoiounndp(airnamnge/tmerLs), PFOA found (ng/mL) - (Peak aresalo-pinetercept) x DF x aliquot factor DAlFiquofatcftaocrtobry-w1h0ich the final volume was diluted, if necessary. Page 6of 7 0531210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 57 o f 65 T ag elO S o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: POOO! 131 Exygen Research Method Number vooo 1785 | ANALVUCAL METHOD Method of Analysis for the DMeatemimmianlaLtiiovnerobfyPeLrCfl/uMorSo/oMcStanoic Acid (PFOA) in Small j 14.2 tFhoerfsoalmlopwliensgfeoqrutiafiteiodnwtoithcaklcnuolwatne tahme opuenrctsenotfrePcFoOveAryp. rior to extraction, use Recovery (%) = [total analyte found (ng/mL) - analyte found in control (ng/mL)} 1QQ analyteadded (ng/mL) 14.3 Use the following equation to convert the amount of PFOA found in ng/mL to ng/g (ppb). PFOA found (ppb) * (PFOA fousnadm(pnlge/mwLei)gxhtf(inga)l volume (mL)l Page 7 o f 7 E0S-0210 Interim R epo rtm * Northern Alabama Potable Water Systems, December 2005 Page 58 o f 65 Page 110 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: P0001131 AMNeAthLodYNTuImCbAerL: MV0E0T01H7O86D Method of Analysis for theMDeatmermmailnSateirounmofbPyeLrfClu/MorSo/oMctSanoic Acid (PFOA) in Small Analytical Testing Facility: SE30txa5yt8egeCRnoeRslleeeagsreeca,hrPcDAhri1v6e801 Approved By: TPaeuchl nCiocanlnoLlelayder, LC-MS, Exygen Research Date / JVSoihclne//P7Frlae?hsiedretynt, __________________ Operations, Exygen Research _D_a_te Total Pages: 7 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 5!) of 65 Page 111 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: POOOl 13 1 Exygen Research Method Number V0001786 MethodofAnalysisfortheMDAeatmNermAmaLilnYSaTteiroIuCnmAofbLPyMeLrECiluT/MoHrSoO/oMDctSanoic Acid(PFOA) inSmall 1.0 Scope TSacphieidsctmbroyemthHeotdirgichisDtPoeetberfceotoermrm(apLnlCocye/MedLSif/qoMuriStdh)eiCnihssormolamatlilaotmnogaamrnadmphqayulasnceotriuutmaptli.eodn otof paerftlaunodroemoctaMnaosics 2.0 Safety 22..21 pCArolewncsaauuyltstiootnhbses.earpvperosapfreialtaebMorSatDorSybperfaocrteicheas.ndling any chemical for proper safely 3.0 Sample Requirement 33..12 3.3 pANsSaratoommljeesppaacllmeste.tspclmloemullpsLetrcototocifoetanselsslpiotnrwsgoaecmidesdptnouleerceefosdomerwdpeixllfeltotrberalecsyteistrophuneam.cwifstiaoemdropiolnemst.hteeHmsoapwmeerpavltieunrrg,efpbroleazfnoenrefosuresrteuh.mis 4.0 Reagents and Standards 44444.....13524 PAMAWemceraeftmthtleouaonronn-iroHtoiruloiPm-lceLtHa-ACnPHcoLegiPctCraLaAtdgCeecr-agiddrAae-d.CeS.iSg.mRae-aAglednritcGhrade 5.0 Instrument and Equipment 5.1 555555555.........5948316270 D52sADA1AAp1o50iL2uimnslsd5vmtmpahpoceLe-loioovypLLmgnscsnitiathciaplddLnscbeebeiieaLalsseptletclrppteqoDeetboomHeurpscPafssdiiPoio(plaapEca1trlLbbperolnnm0ellteoCceeat0dacpaeen-tppnsvit1wopc.aooric0aaewanelli0lyywttpdsh-0ppkaeml(dirrbmiap5tooqalo.0LeupptvuM-aiyyao1tadllhrfafnOeeiosadrnnbOcrsebeeohpua1lStrLcceedt0oplee,ai-emvennnk11socgtt0.a0rtrilt0irniu0toofoft-muupgem20ggLgre0ae.eer0)0pati,ue0ttnthuuwriL0jobbe0()iecnecL.t1shastCa.o.pgdnr/a.MidbscplaqSeopu/MsaaoabnbSflteil)te.paotutifimopinsnp..jiencgtinugp 5-l2o012) 555...111123 VSWPoaErtteevrxasecSru.eupmPmakanViafocl6d.cc (lg) tC18 SPE cartridges. Pag* 2 u p -- E05^0210 InterlnrReportltl8 Northern Alabama Potable Water Systems, December 2005 Page 60 o f 65 ------Tage T f2 of122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: POOOl 131 Exygen Research Method Number VOOO(786 | A lW V nC A L METHOD j Method of Analysis for theMDaetmermmailnSateirounmobfyPeLrCflutMotSo/oMctSanoic Acid (PFOA) in Small 55..1145 CWernistrti-faucgtieoncasphaabkleer.of spinning 15 mL polypropylene tubes at 3000 rpm. 6,0 Chromatographic System 6.1 6666....5423 MGAT(MPenro/oamNabbdlpi:iyilleeet8einr2caPPta5thPhul0aarrC5oses-eo:eg0lr(5(ua3BA2mm0)1)n:3C:::0F)lMu2omepthMhaasnAeomlRmP o(KnieuymstoAnceetSacteieinntiWfica)t,e2r.1 mm x 50 mm. 5p Tim2208el20...O(0.0.50min) %6622655555A %3337755555B FfmloLw/mRiantei 00000.....33333 666...876 RQInuujeancnTttiiiomtanteiV:ono-:l2umP3eema:kin1Au5rtepeasL.-(ceaxntebrneailnsctraenadseadrdtocaalsibmrauticohnacsu5rv0ep. L). TophteimabizoevtehceoHndPiLtiConssysatreemin.tended as a guide and may be changed in order to 7.0 MS/MS System 7.1 MPoFdOeA: .Electrospray Negative MRM mode, monitoring 4 13 -* 369 m/7 for TophteimaibzoevtehecMonSdMitiSonsysstaerme .intended as a guide and may be changed in order to 8.0 Preparation of Solutions 8.1 Mobile Phase 8.1.1 2ammmMonaiummmaocneituamte taoce1t0at0e0imnLwoatfewr aitserp.repared by adding 0.154 g of Alternate volumes may be prepared. Page 3 of t E05-0210 lntertmRep6rt~#18 ~ Northern Alabama Potable Water Systems, December 2005 Page 61 o f 65 Page 113 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: POOOl 131 Exygen Research Method Number V0001786 [ ANAL YT1CAJ- METHOD Method of Analysis for theMDeatmermmailnSateiroanmobfPyeLrfClu/MorSo/oMctSanoic Acid (PFOA) in Small 1J 9.0 Standard Preparation 9.1 9999S....t1111a....n1342darTfmaAommiaPAdnrphrfoeLL1Spe0aea1mt2tpr.hn.oo1o5os10aaatctffx2rhmnoplpketi5ytoechgmg/hLtaFklme/id/emacmoiLasaLtn1ar1tetLnDLl.toe00laidLyscfP0foitfDpko1fcEao4fopg2ranPrsptrg/5bttdioCEmtriif/-oiefolamifmuitcLpbrnaicttcdalaLoLnaieSastortdti.(ottaiLosonlciotloeolanDiounuoo.lnrltutPesnlisfisteoooE.o-iosclnon1ltultubanuet0ttbooidti0otitloooteanflnpneoafofg.siroonff/arpfmiaPrnuPlaeFLarFvliOmttoOooyvAalfA)uboxPmelaiiusmiFnsesmdOptuoopremAderrfeeopil1dpapbfu0areyti10erenre0diwwodta0obedirbwyitegh1yofibh0trfmhrb0iini6gernmmigetnghmreagai1ltnontih0onngowargltmnhiHotaisghnItIl 9.2 Standard Calibration Solutions 9.2.1 9.2.2 pLoTrfChetep/hMaefroSe0ld/.lM1oawpSsignnc/egmaeldiiLsberdfaao.ttriytoipnfiiccsaatatlinoednxaasrmdolspualteiro:enap.dredpitaiorendalincomnectehnatnraotliovnias dilution may be S99oo..C22flou..F34ntoicore11125n1tni000nA.Sa..f000tf000tietrncloaet2gaetdrt/ierimeoCondnanLa.tlto)lec6vaolVCilb(uo,m521r1m1l1ua..u.00000L0ptmei)soteonanssidtxacnmodDnoacnrid(let1um11111hsn00000t0steL000000i.rdn)attoi1o2n5s-mofLsCtLaoDnn(ndcPFge0a20051iE/n.n...r..m152000dtarnlLsaat)miroronawy -bmeopurethpabroetdtleass 10.0 B10a.t1ch 10.2 SEocRienonaetnetcqUhchueepiunbrqentaurmttaracetelhiianottytonessdaffs(soslacaurmobrfanipcentlorlecdon*eltarpeonlxaladtnnsrdalpafciobktteroewdtr)haot(tiosotyrvpuyperniordctijruafeeylpcaltlytp.iecrd2oa0cteecsodoruanrnltaerdolsslrss)epcimkofevouserstrtiwyfiiinefllodclrbutedhateespbaekatcnticlofehiwaesdnt Page 4 of 7 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 62 o f65 Page 114 o f 122 E0S-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: P0001131 Exygen Research Method Number V00017g& | ANALYTICAL m e t h o d ~l Method ofAnalysis for theMDeatmermmailnSateirounmobf yPeLrfClu/MorSo/oMctSanoic Acid (PFOA) in Small 11.0 Sample Extraction 11.1 111111...243 111111...576 11.8 1111..910 11.11 AEmmfADLCdVATCa1Mo5siloreneododlLasuealancapnmrddnytoandatolteodeseywrwbsSnLseifuitxfzetftwhaedeawmriepubodeefrettmiogaoehLlsntdsbehlte1raere1,yeaomaytt-ctmrmprho-shs1.efSemrpu2euepLnotapmlprmhLtlcelptCeeroeaumeesLyiidrcftbfnousolnuoLuedeuensaoasnfgnaestmSitflceieomnpeaHitPato.dprnegfhttcEpinPulntlee-eemaldebd3eLnilncienfeieas0ttdCaoin.ritcatnn0oroihrgtmdfttmrn0waraouooaienpsigadrsnfxdaholpetigpSeaneltmSOerC.whtkraaOusuleenyembfsv(mlCosfeil1LSLona)or2oL.(CPmlg-rflugddElS-i/p,mpenrMNi2aoosclac6m0eepopltSlaydoftiors/vmm2pensMtisiforeszruaLiitpocl2emndSbthu)e)op0gul.a.amLsesyetbtv.mael.DeyaceabsoLienl=olDloft*pe.naeennia2tbersCrcsopnllicameuseftaiaaniupawptLnlrftetgeedorger)ttteiir.itvfescgeuuiot1ltohn-uusg0laltteiaubeulonc.tymmemetttio.uaeoLanbsnngeardmoussrahnfeaI(datSfdsdhukoedraaerrynmtturie3ofmdL5yl 12.0 Chromatography 12.1 12.2 12.3 12.4 12.5 getSvsesiismUaASILhnnoaanxeienatCtsntejummfjancramheeeetssn/eelwrceeecppMutypedsrtrnntllslzabpadelaeetiSdetretdeeincrscheser/ddsra.eaoMetredisessasalenseatfiyesnrssf.oaScbdtAeo,aaspdmlrfttumsroaeasecsPttpsdorttvnmahaaeyihFldeafesnmloeesed.Oinearcdsnrtbypeme.aaaiArsbmlansslnarjehoiptedl-tabat.acouigo1elsronnounyinera0cAnndrntlttstaured.naieasoenorrsiastbsvacdnfntniSmpavlyeasgeoytosncshlepaatlittondo,ibinaclnconedeafuhnrdaoaesrcxaifldandnldarsetcr(aigtsrnsdteoaarbeqoertedsnntentrmeuod.saadtmgsafitucasameunrriscustreoetttrtadtouasisntlntht,uesnssabfditeauesndbtooeairsatbstaelonrtmidlerdfnonotdfiiuohp.ngwlifisurvneltmrieeeecMtcaenedralfusLgoasudsilapsrpnierdistosb1anebdcsemt/nrneroLydxpassdfaonaytroeewrainredrndosrcteedcte.xanssiatebciftlhengadiiol3etebesAaahnetn.tdosrl3wdtabcnyatifnaneeasetyz(irsdagnnogieeodntmainsddtrannrojar.neapedncmefdctqflsiuvipotesanuopiretelroginvalmlirdaveonnnyeksasaalwtds)eonly5.iaaevldnrrh-nateabedhIrIatalIlnaeeae)ssiItl Page 5 of? ---------E 05-02TO m m R e p o ftn n .......Northern Alabama Potable Water Systems, December 2005 Page 63 oj 63 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: POOO1131 Exygcn Research Method Number VOOO1786 _______________________ANALYTICAL METHOD_______________________ Method ofAnalysis for the MDeatmermmailnSateirounmobf PyeLrfClu/MorSo/oMctSanoic Acid (PFOA) in Small 13.0 Acceptance Criteria 13.1 13.2 13.3 dbseCRotM1hanla3fheaemetu0c4rniitgro%roph1kehmvlo3ketedncseasaremoohrtimbtonewoiulgostouuasnarnnl.tofiadnkvb(mfs3Tssaeacb6lmhmomuoeP9enebpFuusatetl4Osrsa.emvo1ttisAanl3usInlehs)ufsoadhopaaratmoeiwttakecpucundeloorlaeddsenpnvspbttataeaebhreyrnlioneeeasndltnktserhgtPsmenotepr-cFhteaefiioeOkratxaareretntlrAierodxesaarfsesalacasypstutlttphlsomeogsitaldnhkfeuno.dtetcvseuossaetrt1rAlbstmbsd0ioieondoeugnyttnenrsrheegetardet-m/amhtebimtoPexae3eixtLtFn6rrrabiOeia,9dctxechceAttatiawhe.esfmnpedpeantrui.enatekhnbi-efoaeelren7oxonL,0mtulre-Oiwatm1wsacQh3itidi0iptbloseea%,lnraI7ettnfhhon0ikeeastf 13.4 wEAranrryorarcnTatleeidbs.tr,atmioanysbtaendeaxrcdlufdoeudndfrotombtehea cstaaltciustliactaiolnouotfliethrebycaulisbirnagtiothnecHuruvgee However, the total number of calibration standards that could be excluded 13.5 13.6 maTsRtp0haeup.nte9serdt9onacnp2troiordoirta(snrReetoexlJtarcitmtesihote0eeend.sp9rs2e8bc0l5oeem%)tev.wfuafosenitcfetIitnfsehbeneectstattoaolt(fiantRbaksdrla)eaamnnrtdifuoposmtnlroeabscnreaesadrdhsloijuobusfluarstslasmtdttaipfiobnnalnedelslastrrrecuodamumusnrutvaiessnelniytjdstezencegotodeetpt.hdnee.edrsarreatiiftotelndimm, iomatrsnue,dsttthhtehabnnee an4a%nalwytiitchainl raunnatnhaelnyttihcealserut nm.uIsft rbeeternetainoanlytizmede.drift exceeds this limit within 14.0 Calculations 14.1 bUgaessneeedtrhaeotenfdopblleyoawtkhienagrMeeaaq)susuasLtiiynongnxttohsoecfatswltcaaunrldeaatperrdtohgceruaarmvme:ou(lnint eoafrPrFeOgrAesfsoiounndp(airnamnge/tmerLs!. PFOA found (ng/mL) " /Peak aresalo-pientercept) x DF x aliquot factor DAFliquofatcftaocrtobry=w2h0ich the final volume was diluted, if necessary. 14.2 Ftiloerfsoalmlopwliensgfeoqrtuiafiteiodnwtoithcaklcnuolwatne tahme opuenrctsenotfrePcFoOveAryp. rior to extraction, use Recovery (%) [total analyte found (ng/mL) - analyte found in control (ng/mL)] ; [(w analyte added (ng/mL)_____________________________ Pgc 6 o f 7 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 200S Page 64 o f 65 Page 116 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Exygen Protocol Number: POOOl 131 Exygeti Research Method Number VQ00I786 | ANALYTICAL m e t h o d ~ Method ofAnalysis for theMDeatmermmailnSateirounmofbPyeLrfClu/MorSo/oMctSanoic Acid (PFOA) in Small 14.3 Uppsbe.the following equation to convert the amount of PFOA found in ng/ml to PFOA found (ppb) " fPFOA fosuanmdptlnegv/omluLmi xe f(imnaLl)volume (mLIl Pig 7 nl"7 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December2005 Page 65 o f 65 Page 117 o f 122 EOS-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Protocol Exygen P0001131; 3M Study Number E05-0210 Amendment 3 Study Title Analysis of Perfluorobutanesulfonate (PFBS), Perfluorohexanesulfonate (PFHS), AND PERFLUOROOCTANESULFONATE ( P F O S ) IN W ATER, SOIL, SEDIMENT, FISH, CLAMS, V eg etatio n, Small Mammal Liver and Small Mammal Serum Using LC/MS/MS for the 3M Decatur Mo nito ring Program PROTOCOL AMENDMENT NO. 3 Amendment Date: May 16, 2005 Performing Laboratory 3M Environmental, Health, and Safety Operations 3M Environmental Laboratory 935 Bush Avenue St. Paul, MN 55106 Laboratory Project Identification E05-0210 E05-0210 GLP Protocol Amendment #3 Page 1of 4 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December2005 Page 118 o f 122 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December2005 Protocol Exygen P0001131; 3M Study Number E05-0210 Amendment 3 This am endm ent m odifies the follow ing portion(s) o f th e protocol: Protocol reads: Testing Facility Exygen Research 3058 Research Drive State College, PA 15801 Phone: (814)272-1039 A mend to read: Testing Facilities Exygen Research 3058 Research Drive State College, PA 15801 Phone: (814)272-1039 3M Environmental Laboratory Building 2-3E-09 935 Bush Avenue St. Paul, MN 55106 , Reason: Addition of 3M Environmental Laboratory as a testing facility allows selected water samples to be sent to the 3M facility for analysis of PFBS, PFHS, and PFOS using the most current version of ETS 8-154 "Determination of Perfluorinated Acids, Alcohols, Amides, and Sulfonates in Water by Solid Phase Extractions and High Performance Liquid Chromatography/Mass Spectrometry". The following modifications to ETS 8-154.1 will be Incorporated into the study: (1) The sample volume extracted and the final SPE elution volume may be adjusted, at the analyst's discretion, to better achieve the detection limits specified in study objectives. As written, ETS 8 154.1 calls for a 40 mL extraction volume with a 5 mL elution volume which results in an eight fold sample concentration. The actual volumes used and the resulting overall concentration factor will be given in the final report. Associated method quality control samples will demonstrate that the volume modifications do not impact method accuracy and precision. (2) A solvent (unextracted) calibration curve may be analyzed with the extracted calibration curve to ascertain extraction efficiency. Samples will be analyzed using the extracted curve unless otherwise stated in the final report. Data from the solvent calibration curve will not be included or discussed in the final report unless it is used to strengthen experimental observations/explanations. (3) ETS 8-154.1 makes no mention of sample surrogate spikes; however, all samples will be spiked with at least one of the following radiolabeled surrogates, PFOA [1,2 13C] (perfluorooctanoic acid), PFOS [1802] and/or PFNA [1,2 13C] (perfluorononanoic acid - C9).The concentration of the surrogate spike may vary depending on the collection event, but typical samples are spiked with a nominal concentration of 0.05 ng/mL. Surrogate spike recoveries will be documented and discussed in the final report. Surrogate recoveries between 10025% will be deemed as meeting method criteria for accuracy. (4) Several method blanks may be prepared and analyzed to better determine a "representative" area count value for the method blank. The method blank area count is instrumental in determining the method limit of quantitation and understanding the average and range of area counts possible is critical. If a method blank value is used as a "zero point" in the final calibration curve, then a brief explanation in the final report or in the raw data as a Note to File must be E05-0210 GLP Protocol Amendment #3 Page 2 of4 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 119 of 122 EOS-0210 Interim Report #18 Northern Alabama Potable Water Systems, December2005 Protocol Bxygen P0001131; 3M Study Number E05-0210 Amendment 3 included to describe why that specific method blank was selected as the zero point (closest to the average value, most conservative (largest) value, etc.) (5) The overall analytical uncertainty for each target (non-surrogate) analyte will be estimated using both the method accuracy and precision. The method accuracy and precision will be calculated using data from laboratory contort spike (LCS) samples prepared and analyzed with the sample set(s). Replicate LCSs must be prepared each day samples and/or other quality control samples are prepared. A sample calculation of how the analytical uncertainty was determined will be provided in the final report and/or raw data. A new revision of ETS 8-154 may incorporate all or several of the modifications listed above. If a new version of ETS 8-154 is issued during the course of this study, sample results must meet the new method requirements regardless if they are listed above. 3M Environmental Laboratory management will approve all documented deviations to the 3M Environmental Laboratory quality system (SOPs, methods, etc.) that occur during the course of this investigation. E 05-0210 GLP Protocol Amendment #3 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 3 of 4 Page 120 of 122 EOS-0210 Interim Report #18 Northern Alebama Potable Water Systems, December 2005 Protocol Exygen P0001131; 3M Study NumberE05-0210 Amendment 3 ' Amendment Approval MichaelA. Santmo, 3M, Sponsor Representative Date William K. Reagan, Ph.D. 3M Environmental Laboratory Manager Date Jaisimha Kesari, Weston Solutions, Study Director E05-0210 GLP Protocol Amendment #3 Page 4 of4 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 121 of 122 05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December2005 3M Confidential Record of Nonconformance / Deviation Study / Project No. E05-0210 Interim Report #18 Deviation type SOP (Check one) 0 Protocol Protocol Requirements: 1. Identification Date(s) of Occurrence: Document Number: ! 12/9/2005-January 26, 2006 Exygen Protol #P0001131 Amendment #3 (3M) Equipment Procedure Method GPO Other: II. Description (3) E T S 8-154.1 m akes no mention o f sample surrogate spikes; however, all samples will be spiked with a radiolabeled surrogate, P F O A [1 ,2 n C], The concentration o f the surrogate spike m a y vary depending on the collection event, but typical sam ples a re spiked with a nom inal concentration o f 0.0 5 ng/mL. Surrogate spike recoveries will be docum ented and discussed in the final report. Actual procedure/process: PFOA[1,2 13C] was not included as a surrogate spike in any of the sample/sample duplicates, or field matrix spikes collected for this project. Surrogate recoveries from the other water projects and previous interim reports for E05-0210 indicate that there are solubility/stability issues with this surrogate in water. The 3M Environmental Lab has started an official investigation (which is not part of E05-0210) to determine the root cause for the low recoveries. Until this situation is fully understood and a method modification has been incorporated to eliminate or circumvent the issue, PFOA[1,2 13C] will not be added as surrogate._____________________________________________________ _ _ _ _ _ _ _ III. Actions Taken _______________________ (such as amendment issued, SOP revision, etc.)______________________ Corrective Action ( Yes El No) Reference: Acceptability of the nonconforming work: Surrogates were originally used as an additional quality control element in conjunction with lab control spikes and field matrix spikes. The surrogate recovery was never intended to be the sole confirmation of that the analytical method was appropriate for the given matrix. Non-inclusion of the surrogate spike has no impact on the quality and integrity of the data presented here................................................... Actions: Halting of Work Client Notification Work Recall Withholding of Report 0 Other: Non-inclusion of the surrogate will be addressed in the final report. Recorded by: Date: Authorization: Date: Sponsor Approval (GLP Protocols): Technical Manager Approval (ithalting of work orwork recall are indicated): Date: Date: IV. Authorization to Resume Work _____ Where halting of work occurred, resumption of work must first be approved by Technical Management Technical Manager Approval: Date: (assigned by Study DirecDtoreovrPiarotjieoctnLeNadoat.t_he_e_n_d o_f_st_ud_y_or_p_ro_je_ct) Protocol Deviation Page 1 of 1 E05-0210 Interim Report #18 Northern Alabama Potable Water Systems, December 2005 Page 122 of 122