Document bay7keagje0YaE3wOX4NVJJnk

3M Company St. Paul, Minnesota Fish Sampling and Analysis for PFOS in Largemouth Bass from the Tennessee River in the Vicinity of the 3M Decatur Facility 3M Company Decatur, Alabama May 2012 11P-0094-7 FISH SAMPLING AND ANALYSIS FOR PFOS IN LARGEMOUTH BASS FROM THE TENNESSEE RIVER IN THE VICINITY OF THE 3M DECATUR FACILITY, DECATUR, ALABAMA May 2012 Prepared for 3M Company St. Paul, MN by WESTON SOLUTIONS, INC. West Chester, PA 19380 W.O. No. 02181.129.122 Section TABLE OF CONTENTS Page 1. INTRODUCTION.......................................................................................................... 1-1 2. MATERIALS AND METHODS.................................................................................. 2-1 3. RESULTS....................................................................................................................... 3-1 4. REFERENCES............................................................................................................... 4-1 APPENDICES APPENDIX A 3M ENVIROMENTAL LABORATORY ANALYTICAL DATA PACKAGES APPENDIX B AXYS LABORATORY ANALYTICAL DATA PACKAGE \\Weston\natacc\FOLDERS.0-9\3M-DECAT\Phase 3 Work Plan\Fish sampling PFOS 2012\2012 Fish Sampling Report.doc i Restoring Resource Efficiency LIST OF TABLES Section Page Table 3-1 Fish Sampling Summary and Catch Per Unit Effort............................................. 3-8 Table 3-2 Fish Morphometries and PFOS Concentrations................................................... 3-9 Table 3-3 Largemouth Bass Filet PFOS Concentration Summary Statistics...................... 3-10 Table 3-4 Surface Water PFOS Concentrations-April 2012................................................3-11 \\Weston\natacc\FOLDERS.0-9\3M-DECAT\Phase 3 Work Plan\Fish sampling PFOS 2012\2012 Fish Sampling Report.doc ii Restoring Resource Efficiency LIST OF FIGURES Section Page Figure 2-1 Fish and Surface Water Sampling Locations-April 2012.....................................2-4 Figure 3-1 Largemouth Bass Filet PFOS Concentrations-April 2012................................... 3-4 Figure 3-2 Largemouth Bass Filet PFOS Concentration Box and Dot Plots........................ 3-5 Figure 3-3 Largemouth Bass Filet PFOS Concentrations vs. Length................................... 3-6 Figure 3-4 Surface Water PFOS Concentrations-April 2012................................................3-7 \\Weston\natacc\FOLDERS.0-9\3M-DECAT\Phase 3 Work Plan\Fish sampling PFOS 2012\2012 Fish Sampling Report.doc iii 1. INTRODUCTION Fish sampling was performed at three adjoining reaches spanning a distance of 2.5 miles in the Tennessee River in the vicinity of the 3M facility in Decatur, Alabama from April 3 through 6, 2012 to characterize current concentrations of perfluorooctanesulfonate (PFOS) in filet samples from largemouth bass (Micropterus salmoides). The largemouth bass was selected for sampling for the following reasons: Correspondence with the fish samples collected in earlier sampling performed by 3M in 2004 and 2006 and subsequent collections by the Alabama Department of Environmental Management (ADEM) in 2007, 2008 and 2009; Predator species representing the highest trophic level; and Recommended target species (EPA, 1993; EPA 2000). In addition to collecting fish samples, surface water samples were collected from each fish sampling reach for PFOS analysis. Prior to sampling, a draft Work Plan for Fish Sampling and Analysis for PFOS in Largemouth Bass in the Tennessee River in the Vicinity of the 3M Decatur Facility (draft Work Plan) was prepared for review by the Alabama Department of Environmental Management (ADEM) and the Alabama Department of Public Health (ADPH). The Work Plan was subsequently revised to reflect comments received (WESTON, 2012). The following sections describe the sampling activities and provide the analytical results for PFOS concentrations in filet tissue and water samples. \\Weston\natacc\FOLDERS 0-9\3M-DECAT\Phase 3 Work PlanVFish sampling PFOS 2012\2012 Fish Sampling Report doc 1-1 Restoring Resource Efficiency 2. MATERIALS AND METHODS A scientific collection permit (SCP) was obtained from the Alabama Department of Conservation and Natural Resources (DCNR) for fish collection. All collection efforts were performed in accordance with the SCP requirements including providing advance notification of pending field collection activities to the appropriate DCNR regional fisheries management/enforcement personnel. ADEM personnel were also notified and observed virtually the entire sample collection process. Electrofishing was conducted using a boat-mounted, generator-powered Smith-Root VI-A electrofisher fitted with two bow-mounted anodes and a single trailing cathode. Sampling reaches were shoreline lengths of approximately 0.75 miles. The upper reach was the southern shoreline of the Tennessee River above the confluence of Bakers Creek and extended to the east. The central reach was the shoreline within the mouth of Bakers Creek and the lower reach extended along the southern shoreline of the Tennessee River adjacent to the 3M property. The mouth of Bakers Creek and lower sampling reaches correspond respectively with the DBC and DOU collection locations in the 2004 and 2006 fish sampling activities performed by 3M (Weston 2008). The three fish sampling reaches are shown in Figure 2-1. Electrofishing focused on shoreline areas where sufficient structure exists to attract and hold largemouth bass. Based on input from the ADEM, the target size range for largemouth bass to be collected was 13-18 inches in length in an attempt to coincide with the size range of fish collected previously by ADEM. The target sample size of largemouth bass from each ofthe three fish sampling reaches was 10 fish for a total target of 30 fish specimens. Fish collection was performed under oversight by personnel from ADEM who were present in a separate boat. These individuals also observed sample preparation and other procedural aspects of the study. Surface water samples were collected using a peristaltic pump from each of the three sampling reaches during fish collection activities for PFOS analyses. Sampling depth was determined on the basis of total water depth. Because the surface water sampling locations were less than 10 feet deep, samples were collected at the 0.6 depth (six-tenths of the total depth of water measured from the surface). In addition to collecting surface water samples \\Weston\natacc\FOLDERS 0-9\3M-DECAT\Phase 3 Work Plan\Fish sampling PFOS 2012\2012 Fish Sampling Report doc 2-1 Restoring Resource Efficiency for laboratory analysis of PFOS concentrations, routine water quality data including the parameters of temperature, specific conductance, dissolved oxygen, pH and turbidity were also collected at the sampling locations. In order to collect data suitable for assessing the range of variability in fish tissue PFOS concentrations and for consistency with past 3M and ADEM collections, samples were prepared from individual fish specimens for analysis rather than composite samples from multiple specimens. Sample IDs were assigned in the sequence of sample collection and did not contain information on the sampling locations. Each fish retained for tissue analysis was weighed and the total length was measured. Per input received from ADEM, skin-off filet tissue samples were prepared in the field from the right side filets of the fish specimens. In addition to the primary samples, field duplicate samples and quality assurance/quality control (QA/QC) samples were prepared. Field QA/QC samples included the collection of three equipment rinseate blanks to document the adequacy of decontamination of knives used to prepare the fish filet samples. Filet tissue samples were shipped frozen to the Michigan State University Wildlife Toxicology Laboratory (MSU WFL) for preparation of tissue homogenates. As instructed by the Study Director, MSU WTL forwarded the homogenized samples to the appropriate laboratories for analysis. The 3M Environmental Laboratory in St. Paul, Minnesota served as the primary laboratory for this study. The AXYS Analytical Services, Ltd. (AXYS) laboratory in Sidney, British Columbia, Canada, provided validation results in this study design. Field duplicate samples were prepared in the field from the left side filets of the chosen fish specimens. The left side filet was removed after the right side filet had been removed and was prepared as a skin-off filet tissue sample. Six left side skin-off fillets were chosen at random and sent as field duplicate samples to MSU WTL for preparation of tissue homogenates. Three of these duplicates were provided to the 3M Environmental Laboratory and the other three were sent to AXYS. The source of the blind field duplicates was not revealed to either laboratory. The blind field duplicates had unique sample identification numbers on the Chain of Custody form sent to the laboratory such that the laboratory could not determine its source. Additional left side filets collected and \\Weston\natacc\FOLDERS 0-9\3M-DECAT\Phase 3 Work PlanVFish sampling PFOS 2012\2012 Fish Sampling Report doc 2-2 Restoring Resource Efficiency not required for QA/QC analysis were prepared as skin-on filets, frozen and transferred to the 3M Environmental Laboratory to be retained indefinitely. Additionally, process control fish samples were sent by the 3M Environmental Laboratory to the MSU WTL to be analyzed before, during and after the processing of the field samples to confirm adequate decontamination of homogenization equipment between preparation of individual study samples and the absence of other contamination vectors associated with the preparation of the tissue homogenates. Following homogenization, these samples were returned to the 3M Environmental Laboratory for analysis. Finally, a standard reference material (SRM) fish tissue sample obtained from the National Institute of Standards and Technology (NIST) was sent to MSU WTL to be repackaged into two aliquots. One aliquot was forwarded to AXYS for analysis and the other aliquot was sent to the 3M Environmental Laboratory for analysis. The 3M Environmental Laboratory analyzed each tissue homogenate sample and a laboratory duplicate for PFOS. As noted above, three split samples prepared from skinoff left side filets (representing 10% of the target sample number collected) were also analyzed for PFOS by AXYS. Surface water samples were analyzed for PFOS by the 3M Environmental Laboratory. \\Weston\natacc\FOLDERS 0-9\3M-DECAT\Phase 3 Work PlanVFish sampling PFOS 2012\2012 Fish Sampling Report doc 2-3  irrr-fr' 3M Decatur ` '. n ; File: Y:\3MDec\MXD\Fish_SW_Sampling_Locs.mxd, 5/1/2012 12:09:12 PM, ricksc LOC001 LOC002 Middle Reach i m LOC003 Bakers \Creek " " ___ Legend: Fish Sampling Location Surface Water Sampling Locations Imagery Source: ESRI Bing Mapping Service 2011 0 500 1,000 Feet Figure 2-1 Fish and Surface Water Sampling Locations April 2012 Bakers Creek and Tennessee River 3M Decatur, AL Facility 3. RESULTS A total of 26 largemouth bass were collected between the three sampling reaches during the week of April 2, 2012. Ten fish each were collected in the upper reach above the mouth of Bakers Creek and the middle reach at the mouth of Bakers Creek and six fish were collected from the lower reach below the mouth of Bakers Creek. A summary of the individual electrofishing runs including the overall elapsed times, the timer seconds recorded during which a potential was applied by the electrofishing unit, the number of bass captured and the catch per unit effort (CPUE) is provided in Table 3-1. Total time of collection varied by reach with the upper reach requiring approximately two hours and the middle reach requiring approximately 3.5 hours to obtain the target number of 10 fish specimens each. In contrast, a total of over 10 hours were expended to obtain the six fish specimens from the lower reach. On April 6, 2012, the last scheduled day for fish collection, efforts to collect additional specimens from the lower reach were suspended when winds of over 20 miles per hour from the north northeast prevented safe vessel operation along the lee shore. Fish lengths, weights and skin-off filet PFOS concentrations are tabulated by sampling reach in Table 3-2 and analytical results are shown in Figure 3-1. The analytical data packages for analyses performed by the 3M Environmental Laboratory are provided in Appendix A and the analytical data package for analyses of split samples performed by the AXYS laboratory is provided in Appendix B. While the majority of the fish were within ADEM's target length range of 13 to 18 inches, one specimen from the middle reach slightly exceeded the target range (MSF13; 18.5 inches total length). One fish in the upper reach (MSF03; 12 inches total length) and three fish from the lower reach (MSF01, MSF16 and MSF22; 10.75, 12.5 and 11.25 inches total length, respectively) were below ADEM's target range but above the 8 inch minimum length established in the original draft Work Plan. Comparisons of primary sample and field duplicate sample PFOS concentrations obtained from analysis by the 3M Environmental Laboratory indicated good correspondence with relative percent difference (RPD) values ranging from 2 to 9% for \\Weston\natacc\FOLDERS 0-9\3M-DECAT\Phase 3 Work PlanVFish sampling PFOS 2012\2012 Fish Sampling Report doc 3-1 Restoring Resource Efficiency the study fish. Comparisons of split sample PFOS concentrations obtained by the AXYS laboratory of left hand filets with the results from the 3M Environmental Laboratory analyses of the corresponding right hand filets indicated less, but acceptable, agreement with RPDs ranging from 21 to 33%. The AXYS PFOS concentrations were higher than the respective 3M Environmental Laboratory PFOS concentrations for the split samples. The AXYS analyses did not include analysis of PFOS laboratory matrix spike (LMS) sample(s) at sufficiently high concentrations relative to endogenous concentrations of PFOS for use in the evaluation of method performance. The 3M Environmental Laboratory analyzed PFOS LMS samples at relevant concentrations for each of the specimens and LMS recoveries ranged from 77% to 125%. (overall mean recovery standard deviation of 99% 10%). The overall comparison of PFOS concentrations measured in all fish samples indicated good agreement between laboratories. In general, PFOS concentrations reported by AXYS were greater than those from 3M for the samples. There was a systematic difference in the reported PFOS concentrations between both laboratories, but this is not unexpected given the potential difference in analytical methodologies used at each laboratory (Lindstrom et al. 2009). For instance, the use of linear isomer PFOS calibration with a 499 to 80 MS/MS transition for data acquisition by AXYS but not by the 3M Environmental Laboratory can bias high the quantification of mixed branched/linear PFOS in environmental samples by 8% (Riddell et al. 2009). Given what is known about the variability in the quantification of PFOS in biological tissues, results from this interlaboratory comparison reinforce the validity of the results of this study. Summary statistics for the largemouth bass filet tissue PFOS concentrations obtained by the 3M Environmental Laboratory analysis of primary (right hand filet) samples are provided in Table 3-3. Median largemouth bass filet tissue PFOS concentrations were 624, 1930 and 979 ng/g for the upper, middle and lower reaches, respectively and 852 ng/g for the combined data set (all reaches). Fish filet concentration data for each of the reaches is depicted in box plots (Figure 3-2). A scatter plot of PFOS and specimen length indicates no correlation between concentration and length (Figure 3-3). \\Weston\natacc\FOLDERS 0-9\3M-DECAT\Phase 3 Work Plan\Fish sampling PFOS 2012\2012 Fish Sampling Report doc 3-2 Restoring Resource Efficiency Only one of three intended sets of water samples were collected. Collection of a second set was planned for April 6, 2012 but, as previously noted, unsafe conditions on the river precluded this action. The intended splitting of water samples with the AXYS laboratory did not occur, in part due to the accelerated pace of this study. Surface water PFOS concentrations and routine water quality parameters are tabulated in Table 3-4 and shown in Figure 3-2. \\Weston\natacc\FOLDERS 0-9\3M-DECAT\Phase 3 Work PlanVFish sampling PFOS 2012\2012 Fish Sampling Report doc 3-3  irrr-fr' 3M Decatur Lower Reach MSF01 MSF16 MSF17 MSF28 (MSF17D) MSF24 MSF24D (AXYS) MSF25 MSF26 MSF29 (MSF26D) P F S (ng/g) 4770 1520 165 180 437 537 306 2090 2050 LOC001 LOC002 A Middle Reach Middle Reach MSF02 MSF07 MSF08 MSF09 MSF27 (MSF09D) MSF10 MSF11 MSF11D (AXYS) MSF11D (AXYS Dup) MSF12 MSF13 MSF14 MSF15 PFOS (ng/g) 4240 2920 1360 2020 2120 1840 2940 3720 3510 526 950 3370 754 Upper Reach MSF03 MSF04 MSF05 MSF05D (AXYS) MSF06 MSF18 MSF19 MSF20 MSF21 MSF22 MSF23 PFOS (ng/g) 70.7 576 471 656 156 701 675 1180 671 4660 52.5 LOC003 File: Y:\3MDec\MXD\Largemouth_Bass_PFOS_April2012.mxd, 5/9/2012 9:56:43 AM, ricksc _ r . . . r. Bakers \Creek Legend: Fish Sampling Location Surface Water Sampling Locations Note: PFOS concentrations are the average of primary and duplicate sample analytical results on a wet weight basis. Imagery Source: ESRI Bing Mapping Service 2011 S 0 500 1,000 Feet Figure 3-1 Largemouth Bass Filet PFOS Concentrations April 2012 Bakers Creek and Tennessee River 3M Decatur, AL Facility Figure 3-2 Largemouth Bass Filet PFOS Concentration Box Plots 5000.0 4000.0 PFOS (ng/g) 3000.0 2000.0 1000.0 0.0 -----1---------------- 1---------------- 1---------------- 1--- 1 Combined Lower Middle Upper Reach W h is k e rs (T ) d e n o te th e la rg e s t (a n d s m a lle s t) a d ja c e n t v a lu e s w ith in 1 .5 tim e s th e in te rq u a rtile ra n g e (IQ R ) R e d d o t d e n o t e s a n o u t l i e r g r e a t e r t h a n 1 . 5 t i m e s t h e I Q R a b o v e t h e u p p e r q u a r t i l e ( 7 5 th p e r c e n t i l e ) v a l u e Figure 3-3 Largemouth Bass PFOS Concentrations vs. Length Upper Middle A Lower 3M Decatur Sam ple ID LOC001 PFOS (ng/mL) 0.450 Sam ple ID LOC002 PFOS (ng/mL) Sam ple ID LOC003 PFOS (ng/mL) 0.0237 Middle Reach File: Y:\3MDec\MXD\Surface_Water_PFOS_April2012.mxd, 5/10/2012 10:45:30 AM, ricksc Bak ICre Fish Sampling Location Surface Water Sampling Locations PFOS concentrations are the average of primary and duplicate sample analytical results. Imagery Source: ESRI Bing Mapping Service 2011 S 500 1,000 Feet Figure 3-4 Surface Water PFOS Concentrations April 2012 Bakers Creek and Tennessee River 3M Decatur, AL Facility Table 3-1 Fish Sampling Summary and Catch Per Unit Effort Date 4/3/2012 4/3/2012 4/3/2012 4/3/2012 4/4/2012 4/4/2012 4/4/2012 4/4/2012 4/4/2012 4/5/2012 4/5/2012 4/5/2012 4/6/2012 Run 1 2 3 4 1&2 3 4 5 6 1 2 3 1 Reach Lower Middle Upper Middle Lower Upper Upper Lower Lower Lower Lower Lower Lower Hours 1.00 1.00 1.00 1.08 2.00 1.25 1.20 1.00 1.00 1.17 1.25 2.00 0.67 Timer (seconds) 1355 1680 1939 1897 3459 2253 1112 1809 1525 2071 2020 2806 1291 Timer (hours) 0.38 0.47 0.54 0.53 0.96 0.63 0.31 0.50 0.42 0.58 0.56 0.78 0.36 Summary Lower Reach Middle Reach Upper Reach Total 9 2 3 Total Hours 10.08 2.08 3.45 Total Timer (seconds) 16336 3577 5304 Total Timer (hours) 4.54 1 1.47 Overall 14 15.62 25217 7.00 Notes: CPUE = Catch per unit effort Timer seconds record the number of seconds that the electrofisher potential was applied Bass (count) 1 1 4 9 2 2 4 0 0 1 0 1 1 Total Bass (count) 6 10 10 26 CPUE (bass/hour) 1.00 1.00 4.00 8.31 1.00 1.60 3.33 0.00 0.00 0.86 0.00 0.50 1.50 Average CPUE (bass/hour) 0.61 4.65 2.98 1.78 CPUE (bass/timer hours) 2.66 2.14 7.43 17.08 2.08 3.20 12.95 0.00 0.00 1.74 0.00 1.28 2.79 Average CPUE (bass/timer hours) 1.32 9.61 7.86 4.10 Sample ID MSF03 MSF04 MSF05 MSF05D MSF06 MSF18 MSF19 MSF20 MSF21 MSF22 MSF23 MSF02 MSF07 MSF08 MSF09 MSF09D (MSF27) MSF10 MSF11 MSF11D MSF11D (Duplicate) MSF12 MSF13 MSF14 MSF15 MSF01 MSF16 MSF17 MSF17D (MSF28) MSF24 MSF24D MSF25 MSF26 MSF26D (MSF29) Date 4/3/2012 4/3/2012 4/3/2012 4/3/2012 4/3/2012 4/4/2012 4/4/2012 4/4/2012 4/4/2012 4/4/2012 4/4/2012 4/3/2012 4/3/2012 4/3/2012 4/3/2012 4/3/2012 4/3/2012 4/3/2012 4/3/2012 4/3/2012 4/3/2012 4/3/2012 4/3/2012 4/3/2012 4/3/2012 4/4/2012 4/4/2012 4/4/2012 4/5/2012 4/5/2012 4/5/2012 4/5/2012 4/5/2012 Table 3-2 Fish Morphometrics and PFOS Concentrations Analytical Laboratory 3M Environmental Laboratory 3M Environmental Laboratory 3M Environmental Laboratory AXYS Laboratory 3M Environmental Laboratory 3M Environmental Laboratory 3M Environmental Laboratory 3M Environmental Laboratory 3M Environmental Laboratory 3M Environmental Laboratory 3M Environmental Laboratory Sampling Reach Upper Upper Upper Upper Upper Upper Upper Upper Upper Upper Upper Length (inches) 12.00 14.75 17.75 H 14.00 15.75 14.75 14.25 17.00 11.25 17.50 3M Environmental Laboratory 3M Environmental Laboratory 3M Environmental Laboratory 3M Environmental Laboratory 3M Environmental Laboratory 3M Environmental Laboratory 3M Environmental Laboratory AXYS Laboratory AXYS Laboratory 3M Environmental Laboratory 3M Environmental Laboratory 3M Environmental Laboratory 3M Environmental Laboratory 3M Environmental Laboratory 3M Environmental Laboratory 3M Environmental Laboratory 3M Environmental Laboratory 3M Environmental Laboratory AXYS Laboratory 3M Environmental Laboratory 3M Environmental Laboratory 3M Environmental Laboratory Middle Middle Middle Middle Middle Middle Middle Middle Middle Middle Middle Middle Middle Lower Lower Lower Lower Lower Lower Lower Lower Lower 14.50 16.00 13.50 13.25 H 14.00 15.375 H H 15.50 18.50 17.50 17.50 10.75 12.50 13.75 H 16.00 H 16.50 15.00 H Weight (pounds) 0.9 1.7 2.9 H 1.5 2.5 1.8 1.0 2.8 0.8 3.1 1.8 2.2 1.3 1.2 H 1.7 2.0 H H 2.1 3.3 3.6 3.3 0.5 1.0 1.2 H 2.1 H 2.1 1.7 H PFOS Concentration (ng/g) 70.7 576 471 656 156 701 675 1180 671 4660 52.5 4240 2920 1360 2020 2120 1840 2940 3720 3510 526 950 3370 754 4770 1520 165 180 437 537 306 2090 2050 Notes: PFOS concentrations are the average of the primary and duplicate analytical results expresssed in ng/g (ppb) on a wet weight basis Sample IDs with a "D" suffix are field duplicates obtained from left side filet samples Parameter Minimum Maximum Mean Median Geometric Mean Standard Deviation Table 3-3 Largemouth Bass PFOS Concentration Summary Statistics Upper Reach 52.5 4660 921 624 440 1358 Middle Reach 526 4240 2092 1930 1725 1238 Lower Reach 165 4770 1548 979 833 1753 Notes: PFOS concentrations in ng/g (wet weight). Summary statistics obtained from primary data (field duplicate / split sample data not included). Combined Reaches 52.5 4770 1516 852 863 1451 Table 3-4 Surface Water Data Sample Station ID Northing (Decimal degrees) Westing (Decimal degrees) Date Time Temperature (C) Specific conductance (ps/cm) Dissolved oxygen (mg/L) pH (S.U.) Turbidity (NTU) Average PFOS (ng/mL) Lower Reach LOC001 34.644 87.035 04/05/12 9:50 21.63 189 7.74 7.33 35.2 0.450 Middle Reach LOC002 34.638 87.029 04/05/12 10:17 21.77 187 9.75 8.13 23.5 0.691 Upper Reach LOC003 34.637 87.021 04/05/12 10:37 21.5 193 8.13 7.56 21.5 0.0237 Notes: Average PFOS concentration is the average o f primary and duplicate analytical results in ng/mL (ppb) 4. REFERENCES EPA. 2000. Guidance for Assessing Chemical Contaminant Data for Use in Fish Advisories, Volume I, Fish Sampling and Analysis. Third Addition. EPA. Office of Water. Washington, District of Columbia. EPA. 1993. Fish Field and Laboratory Methods for Evaluating the Biological Integrity o f Surface Waters. EPA. Environmental Monitoring Systems Laboratory. Cincinnati, OH. Reynolds. J.B. 1996. Electrofishing. Pages 221-254 in B.R. Murphy and D.W. Willis editors. Fisheries Techniques, Second Edition. American Fisheries Society. Bethesda, MD. WESTON 2008. Perfluorinated Sulfonates Data Report, 3MDecatur, Alabama Facility, Site-Related Environmental Monitoring Program, September 2008, Weston Solutions, Inc., West Chester, PA. Lindstrm G., Karrman A., and van Bavel B. (2009). Accuracy and precision in the determination of perfluorinated chemicals in human blood verified by interlaboratory comparisons. J. Chromatogr. A 1216: 394-400. Riddell N, Arsenault G., Chittim B., Martin JW., McAlees A., and McCrindle R. (2009). Branched perfluorooctane sulfonate isomer quantification and characterization in blood serum samples by HPLC/ESI-MS/(/MS). Environ. Sci. Technol. 43: 7902-7908. WESTON. 2012. Amended Work Plan for Fish Sampling and Analysis for PFOS in Largemouth Bass in the Tennessee River in the Vicinity o f the 3M Decatur Facility. Weston Solutions, Inc., West Chester, PA. \\Weston\natacc\FOLDERS 0-9\3M-DECAT\Phase 3 Work Plan\Fish sampling PFOS 2012\2012 Fish Sampling Report doc 4-1 APPENDIX A 3M ENVIRONMENTAL LABORATORY ANALYTICAL DATA PACKAGES Final Report Concentrations of PFOS in Fish Fillet Tissues from Larqemouth Bass Collected in the Tennessee River in April 2012 Study Title Analytical Study IS 0 1 2-01-01-01: Analysis o f PFOS from Tennessee River Fish - Bakers Creek Area: April 2012 Data Requirement ISO17025 Project Director Charles Young Weston Solutions, Inc. Principal Investigator Cleston C. Lange, Ph.D. 3M Environmental Laboratory Report Completion Date May 02, 2012 Analytical Laboratory 3M Environmental Health and Safety Operations Environmental Laboratory 3M Center, Bldg 26CF05-N-17 St. Paul, MN 55144 3M LIMS Project Identification IS 0 12-01-01-01 Total Number of Pages 25 SW ACCREDITED Testing Certificate #2052.01 This laboratory maintains A2LA accreditation to: ANS1/1SO/1EC17025:2005fo r the specific tests/calibrations listed in A2LA Certificate # 2052.01 and meets the principles o fISO 9001:2000. The tests results included in this report are covered by this accreditation following methodETS-8-045.1 Study: IS012-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 Table of C o n ten ts Table of Contents.......................................................................................................................................2 List of T ables...............................................................................................................................................3 1 Study Information.................................................................................................................................4 2 Sum m ary..............................................................................................................................................5 3 Introduction...........................................................................................................................................7 4 Reference Substances....................................................................................................................... 8 5 Method S um m ary................................................................................................................................8 5.1 M ethods...................................................................................................................................8 5.2 Sample Receipt.......................................................................................................................9 5.3 Sample Preparation.................................................................................................................9 5.4 LC/MS/MS Analysis...............................................................................................................10 6 Analytical Results...............................................................................................................................12 6.1 Calibration..............................................................................................................................12 6.2 Limits of Quantitation (LO Q s)...............................................................................................13 6.3 Continuing Calibration...........................................................................................................14 6.4 Blanks.....................................................................................................................................14 6.5 Laboratory Control Spikes (LCSs)........................................................................................14 6.6 Laboratory Matrix Spikes (LM Ss)........................................................................................ 19 6.7 NIST SRM 1947 Results...................................................................................................... 20 6.8 Individual Sample Results.................................................................................................... 21 7 Conclusion..........................................................................................................................................23 8 Data/Sample Retention.................................................................................................................... 24 9 List of Attachm e n ts ............................................................................................................................24 10 Signatures..........................................................................................................................................24 2 Study: IS012-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 L ist of Tables Table 1. PFOS Results for Tennessee River Largemouth Bass FilletTissues, April 2012.................. 6 Table 2. Analytical Reference Substances............................................................................................. 8 Table 3. Instrument Information..............................................................................................................11 Table 4. Gradient Liquid Chromatography Conditions (ETS-8-045)................................................... 11 Table 5. Mass Transitions.......................................................................................................................11 Table 6. Analytical Batches for PFOS in Tennessee River Fish (April2012)...................................... 12 Table 7. L-PFOS Solvent Calibration Results....................................................................................... 13 Table 8. L-PFOS LCSs and Matrix Blank Results.................................................................................15 Table 9. brPFOS LCSs and Matrix Blank Results................................................................................ 17 Table 10. Homogenization Process Control Results............................................................................ 20 Table 11. Individual PFOS Results for Fillet Tissues from Fish Collected in the Tennessee River, April 2012............................................................................................................................... 21 3 Study: IS012-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 1 Study Information Sponsor 3M Company Project Requestor Gary Hohen stein Environmental Manager Special Projects, EHS Operations Bldg 224-5W-03 St. Paul, MN 55144 Phone:(651)737-3570 aahohenstein@mmm.com Sampling Coordinator Charles Young Weston Solutions, Inc. 1400 Weston Way West Chester, PA 19380 610-701-3787 charles.vouna@westonsolutions.com Principal Analytical Investigator Cleston Lange, Ph.D. 3M Environmental Laboratory 3M Center, Bldg 260-5N-17 St. Paul, MN 55144 Phone: (651)-733-9860 clanae@mmm.com Analytical Testing Facility 3M Environmental Laboratory Building 260-5N-17 St. Paul, MN 55144 Study Personnel (3M Environmental Laboratory) Marlene Heying, Analyst (Pace Analytical, Professional Services) Kelly Ukes, Analyst (Pace Analytical, Professional Services) Jon Steege, Analyst (Pace Analytical, Professional Services) William Reagen, Ph.D., 3M Environmental Laboratory Management Study Dates Study Initiation: April 10, 2012 Interim Analytical Initiation (PFOS): April 16, 2012 Interim Analytical Completion (PFOS): April 24, 2012 Interim Report Completion: April 30, 2012 4 Study: IS012-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 2 Summary This analytical report provides the perfluorooctane sulfonate (PFOS) concentrations measured for twenty-nine (29) homogenized fillet tissues (skin removed) that were obtained from twenty-six (26) largemouth bass collected from the Tennessee River in April 2012 near Decatur, Alabama. This analysis was conducted in support of the Weston Solutions, Inc. project plan, titled "Work Plan for Fish Sampling and Analysis for PFOS in Largemouth Bass from the Tennessee River in the Vicinity of the 3M Decatur Facility, Decatur, Alabama". The analytical aspects reported herein were conducted in the 3M Environmental Laboratory as 3M study IS 012-01-01-01. The 26 largemouth bass were collected from the Tennessee River in early April 2012 and were filleted in the field (skin removed) by Weston Solutions, Inc. personnel. The 26 right side fillets from each fish, and 6 left side fillets from six random fish as blind field duplicates, were sent to the Michigan State University W ldlife Toxicology Laboratory (MSU-WTL) for homogenization. The processed fish homogenates from 26 right side fillets and 3 of the left side fillets were then packaged in baggies and shipped frozen with 3 process controls and 1 standard reference material (NIST SRM 1947) to the 3M Environmental Laboratory for quantitative analyses of PFOS. The other 3 left side fillet homogenates (skin removed), along with one aliquot of NIST SRM 1947, were sent to Axys Labs for analysis of PFOS as independent verification of the 3M Environmental Laboratory results. The remaining 20 whole fillets, skin on and not homogenized, were sent directly to the 3M Environmental Laboratory by Weston Solutions, Inc. field personnel to be retained frozen as "archive" samples for potential future use. The twenty-nine (29) homogenized fish fillet tissues were received on April 14, 2012 and April 17, 2012 as two separate shipments. The analytical laboratory was blind to the true identities of the Tennessee River fish with regard to sex, age and specific sampling location, which was only known to Weston Solutions, Inc. and were only labeled with unique, generic IDs. In addition to the largemouth bass tissues, three (3) process control fish tissues (pre-study, mid-study and post-study controls) and one (1) standard reference material (NIST SRM 1947; Lake Michigan fish tissue) were received with the fish samples from MSU-WTL. Those control materials were previously provided by the 3M Environmental Laboratory to MSU-WTL prior to fish collection and processing to serve as controls for evaluating the homogenization process and evaluating the PFOS results relative to a NIST SRM 1947 fish tissue PFOS reference value. In contrast to Tennessee River fish samples, the four control tissues were identifiable by the laboratory in order to evaluate them for quality assurance purposes, as discussed herein. Once received at the 3M Environmental Laboratory, samples were logged into the electronic laboratory information management system (LIMS) where each sample was assigned a unique 3M LIMS sample ID under project IS012-01-01-01. Fish homogenates were stored frozen until preparation for analysis. Fish homogenates were re-homogenized, extracted by protein precipitation method and analyzed by liquid chromatography with triple quadrapole mass spectrometric detection (LC/MS/MS) following the validated and published 3M Environmental Laboratory analytical method ETS-8-045.1. Quantitation was performed by internal standard (IS) method using stable isotope labeled ' 'Ce-PFOS as the IS and utilizing linear isomer of PFOS (L-PFOS) solvent calibration. Each sample was extracted in duplicate along with corresponding laboratory matrix spikes (LMSs) to evaluate sample-specific PFOS recovery. As additional quality control during the analysis, each sample received a fixed quantity of a stable isotope labeled surrogate reference standard (SRS) l;,C4-PFOS prior to extraction; results of which were used as a secondary measure of PFOS recovery from each sample. Laboratory control spikes (LCSs) were employed to evaluate the method performance. Two sets of laboratory control spikes (LCSs) were prepared with each analytical batch in a control bass fillet homogenate; each set with three levels of PFOS fortification at nominal 10.0, 100 and 1000 ng/g, each level prepared in triplicate. One set of LCSs was prepared with L-PFOS, and the second was prepared with a mixed linear plus branched isomer PFOS (brPFOS) to evaluate potential isomer effects on quantitation. Additionally, for analytical batches which contained diluted samples, high-level LCSs were similarly diluted to incorporate some measure of dilution accuracy and precision into the method performance evaluations. The LCS results were used for determining the method performance during 5 Study: IS012-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 the study. The LCS (L- PFOS) results are reported in Table 8 and LCS (br PFOS) results are shown in Table 9 and demonstrated excellent analytical method performance with accuracies of within 100 + 25% for all LCS results and RSDs < 20%. Statistical evaluation of the LCS results provided method uncertainties (95% Cl) of + 19% for L-PFOS and + 11% for brPFOS. The concentrations of PFOS in pre-study, mid-study and post-study homogenization process controls were determined at 0.815 ng/g, 0.909 ng/g and 0.912 ng/g respectively. Those values compared well to the determined 0.804 ng/g average PFOS concentration for the same control tissue which did not travel to MSU-WTL and was not homogenized by MSU-WTL. The RPDs between results for the unprocessed versus MSU-processed control tissues were less than 15% for each and showed that PFOS was not significantly introduced to samples as a result of any sample contamination during processing or shipping of the fish tissue sam pies from MSU-WTL to the laboratory. The average determined PFOS concentration for the duplicate prepared analytical samples are shown in Table 1, along with the RPD of the duplicate results for each. The measured concentrations of PFOS ranged from 52.5 ng/g to 4770 ng/g. The recoveries of fortified PFOS from the LMS prepared for each at a level comparable to the endogenous PFOS level in the corresponding Tennessee River fish sample ranged from 76.7% to 125%, with an overall mean recovery and standard deviation of 98.9% + 10%, as shown in Table 1. The overall recovery of PFOS demonstrated excellent sample specific PFOS measurement accuracy. Additionally, the precision of each result was excellent with RPDs less than 20%, and typically less than 10%. The recoveries of the added stable isotope labeled SRS (13C4PFOS) were within 100 + 20% for all samples, and with excellent precision (RPDs < 20%) for all sam pies, as shown in Table 11. The measured PFOS concentration for a NIST SRM 1947 was 6.71 ng/g (4.0% RPD, 95.3% LMS recovery), and was accurate to 13% RPD with respect to the published SRM 1947 mean reference value and standard deviation of 5.90 ng/g. Table 1. PFOS Results for Tennessee River Largemouth Bass Fillet Tissues, April 2012 3M LIMS ID IS012-01-01-01-021 IS012-01-01-01-022 IS012-01-01-01-023 IS012-01-01-01-024 IS012-01-01-01-025 IS012-01-01-01-026 IS012-01-01-01-027 IS012-01-01-01-028 IS012-01-01-01-029 IS012-01-01-01-030 IS012-01-01-01-031 IS012-01-01-01-032 IS012-01-01-01-033 IS012-01-01-01-034 IS012-01-01-01-035 IS012-01-01-01-036 IS012-01-01-01-037 IS012-01-01-01-038 IS012-01-01-01-039 IS012-01-01-01-040 IS012-01-01-01-041 IS012-01-01-01-042 Sample ID Process Control#1 (Pre-Study) Process Control #2 (Mid-Study) MSF01 120403 MSF02 120403 MSF03 120403 MSF04 120403 MSF05 120403 MSF06 120403 MSF07 120403 MSF08 120403 MSF09 120403 MSF10 120403 MSF21 120404 MSF22 120404 MSF23 120404 MSF24 120405 MSF25 120405 MSF26 120405 MSF27 120403 MSF28 120404 MSF29 120405 Process Control #3 (Post Study) Avg. PFOS Concentration (ng/g)[bl 0.815 0.909la| 4770 4240 70.7 576 471 156 2920 1360 2020 1840 671 4660 52.5 437 306 2090 2120 180 2050 0.912 RPD (%) 2.0% 34% 1.4% 5.3% 1.9% 6.5% 4.9% 5.4% 1.6% 0.13% 5.2% 10% 6.1% 3.5% 0.0056% 12% 3.8% 1.0% 5.4% 9.2% 0.063% 16% LMS PFOS Recovery 102% 99.1% 95.5% 92.4% 81.0% 108% 100% 75.7% 98.7% 99.8% 104% 105% 85.0% 99.5% 99.7% 90.7% 125% 94.2% 105% 124% 102% 102% 6 Study: IS012-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 Table 1. PFOS Results for Tennessee River Largemouth Bass Fillet Tissues, April 2012 3M LIMS ID Sample ID Avg. PFOS Concentration (ng/g)[bl RPD (%) LMS PFOS Recovery IS012-01-01-01-043 NISTSRM1947 6.71 4.0% 95.3% IS012-01-01-01-044 MSF11 12-04-03 2940 3.9% 93.5% IS012-01-01-01-045 MSF 12 12-04-03 526 4.2% 101% IS012-01-01-01-046 MSF 13 12-04-03 950 1.7% 99.0% IS012-01-01-01-047 MSF 14 12-04-03 3370 0.50% 102% IS012-01-01-01-048 MSF 15 12-04-03 754 9.6% 99.4% IS012-01-01-01-049 MSF 16 12-04-04 1520 3.2% 86.9% IS012-01-01-01-050 MSF 17 12-04-04 165 0.75% 84.2% IS012-01-01-01-051 MSF 18 12-04-04 701 2.7% 103% IS012-01-01-01-052 MSF 19 12-04-04 675 0.86% 104% IS012-01-01-01-053 MSF 20 12-04-04 1180 0.82% 104% Notes T he a verage result and relative p ercent d ifference (R PD ) w a s d eterm ined from duplicate analytical sa m ples for each fish hom ogenate A ll reported concentration results are rounded to th re e significant figures w h ile statistical values are rounded to two significant figures T h e a n a ly tic a l m e th o d d a ta u n c e rta in ty w a s 1 1 % fo r b rP F O S (30 6 % b ra n c h e d is o m e r) a n d +, 19% fo r L -P F O S (> 9 9 % linear isom er) based on LC S results. [a] RPD > 2 0 % [b ] T he P F O S resu lt is th e cu m u la tive co n ce ntra tio n o f branched a n d lin e a r iso m ers o f P FO S 3 Introduction A scientific investigation of PFOS levels in fish and surface water collected from the Tennessee River near the 3M Decatur facility was conducted in the Spring of 2012 per the study work plan titled: "Work Plan for Fish Sampling and Analysis for PFOS in Largemouth Bass from the Tennessee River in the Vicinity of the 3M Decatur Facility, Decatur, Alabama". The analytical aspects reported herein are for the analyses of the fish tissues as conducted by the 3M Environmental Laboratory under the 3M study IS O I2-01-01-01. The overall purpose of this study was to provide additional site-specific information on PFOS level in fish and surface waters of the Tennessee River near the confluence of Baker's Creek, located near the 3M Decatur facility. This report describes the sample receipt, extraction and quantitative LC/MS/MS analysis procedures performed for the measurement of PFOS in the fish tissues harvested from largemouth bass collected in early April of 2012 from the Tennessee River, and provides the PFOS concentration results for those samples. Additionally included in this report are the assessment of the precision and accuracy of the results based on several parameters: (1) The precision of sample results was determined from preparation and analysis of duplicate analytical samples for each homogenate: (2) the recovery of PFOS from laboratory matrix spikes (LMS) samples prepared for each sample was a measure of samplespecific accuracy: (3) two LCS sets were prepared with each analytical batch, one fortified with L-PFOS and one with br-PFOS in a control largemouth bass fillet homogenate (Osage Catfisheries, Inc.) to evaluate analytical method precision and accuracy; and (4), the recovery of a stable isotope labeled SRS (13C4-PFOS) from each sample was determined and used as a secondary measure of samplespecific analyte recovery, and accuracy and precision. Personnel from Weston Solutions, Inc were responsible for coordinating all field aspects of the study such as site selection, site parameters, fish and water collection procedures, handling and storage of collected specimens, documentation of individual fish information, filleting, and shipment of intact fish fillets to the MSU-WTL for homogenization. The MSU-WTL personnel were responsible for proper homogenization of fillets, relabeling and distribution of homogenates to the appropriate analytical laboratories. The 3M Environmental laboratory was responsible for ensuring samples were received 7 Study: IS012-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 appropriately, stored frozen and prepared and analyzed for PFOS. In addition, Axys Laboratories was to receive a sub-set of duplicate fish tissue homogenates for 3 of the Tennessee River fish (10% split) from MSU-WTL and one standard reference material (NIST SRM 1947) for independent verification of the PFOS results attained by 3M, and will be reported separately. This was a laboratory-blinded study and the fish ages, sizes and specific river collection locations were not provided to the analytical laboratories. Tennessee River fish samples were only identifiable at the laboratory by coded IDs provided to them by Weston Solutions, Inc. personnel. Weston Solutions, Inc. is responsible for all final analysis of the data for associating PFOS concentrations with collection location, fish species, age, weight, etc. This report focuses solely on the extraction and quantitative LC/MS/MS analysis performed at the 3M Environmental Laboratory and reports the determined concentration results for PFOS in the fish tissues, along with a measure of analytical accuracy and precision of the results based on several quality control aspects. 4 Reference Substances The reference substances used during the analysis of the fish tissues and the consumable supplies are listed in Table 3. Table 2. A n a ly tic a l R eference S ubstances Reference Substance Chemical Name Chemical Formula Use Source expiration Date Storage Conditions Chemical Lot TCR Number P F O S (linear isom er) Potassium Perfluoro-n-octane Sulfonate C,Fl7SO,K Calibration, LMSs & LCSs Wellington Laboratories 10/18/2018 Frozen LPFOSKBM06 TCR08-0001-1/1 PFO S (linear + branched Isomers) Potassium Perfluorooctane Sulfonate C,Fl7SO,K Linear -(-Branched Isomers LCSs Wellington Laboratories 12/1/2014 Frozen br-PFO SKllll TCR11-0041-1/19, -7/19 M 4-PFO S I3Ci -PFOS " C iUCiFl7SO,Na Surrogate Wellington Laboratories 12/21/2014 Frozen MPFOS1211 TCR12-0003-1/14 M 8-PFO S " Cj-PFOS l l C,Fl7SO,Na Internal Standard (IS) Wellington Laboratories 6/17/2012 Frozen M8PFOS0609 TCR09-0051-9/15 Physical Description White Powder White Powder Methanol Solution [a) Methanol Solution [a] Purity 99.8% >98% >98% >98% [a] S ta b le isotope labeled m ate ria ls a re p ro vid e d as m eth a n o l solutions b y th e v e n d o r (sou rce ), th e ve n d o r's reported co n ce ntra tio n is used fo r calcu la tin g co n ce ntra tio n s in sam ples 5 Method Summary 5.1 Methods The re-homogenization of fish tissue homogenates from MSU-WTLs, and the sample extractions and quantitative LC/MS/MS analyses were performed following validated and previously published 3M Environmental Laboratory method ETS-8-045.1 [Malinsky ef. al. 2011, Analytics Chemica Acta 683:pp248-257]. Sample analyses for fish samples occurred from April 17th through April 24, 2012. The analytical runs are identified by the initial of the instrument name used and the date and a letter describing the order of the runs on that day (i.e. j120417a describes instrument Jonas on April 17, 2012 and was the first analytical run of the day on that instrument; j120417b would describe the second run of the day on that same instrument). The identities of the instruments used are shown in Table 3. A summary of the analytical runs for reporting is shown in Table 6. 8 Study: IS012-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 5.2 Sample Receipt On April 12, 2012 the 3M Environmental Laboratory received twenty (20) frozen whole fillets (skin on, not homogenized) directly from the field as "archive" samples which were stored frozen upon receipt. Those samples were received with 3M chains of custody (COCs) # 19436 and #19437. The archive samples were not analyzed as part of this study and were retained frozen for potential future use, but associated with this study. The whole fillet samples were logged into the laboratory LIMS system and consecutively numbered based on their order on the COCs as IS012-01-01-01-001 through -020. On April 14, 2012 nineteen (19) homogenized fillets and two process controls (pre-study and mid-study controls) were received at the 3M Environmental Laboratory from MSU-WTL with COCs #14933 and #14940 and one MSU-WTL COC for the process controls. On April 17, 2012 an additional ten (10) Tennessee River fish tissue homogenates were received with 3M COC #14934, and the post-study process control and NIST SRM 1947 material were received with a MSU-WTL COC. The three process control tissue homogenates (3 ~10-25 gram aliquots of a control bluegill fillet homogenate TN11-0184-1/1) had been provided to MSU-WTL earlier by the 3M Environmental Laboratory for quality assurance purposes to monitor potential sample contamination during the homogenization process. Additionally, two ~5-10 gram aliquots of a National Institutes of Standards and Technology (NIST) standard reference material SRM 1947 (Lake Michigan fish tissue homogenate) was also provided to MSU-WTL, to be re-packaged and one aliquot sent to each laboratory for analysis. The twenty-nine Tennessee River fish tissue homogenates, three process controls and one NIST SRM 1947 reference were logged into the Laboratory's electronic LIMS system in the consecutive order they were received, and in the order listed on the chains of custodies as IS012-01-01-01-021 through ISO12-0101-01-053. 5.3 Sample Preparation Each frozen fish tissue homogenate was removed from its polypropylene sample bag and re homogenized with dry ice, then transferred back into the same sample bag and left to stand overnight in a refrigerator to allow the dry ice to sublime (dissipate). Then for each sample, duplicate nominal 0.5 gram aliquots of tissue homogenate were each accurately weighed into a centrifuge tube for extraction. Additionally, a third and fourth aliquot of each was aliquoted and one fortified with L-PFOS at nominal 50 ng/g and the other at 500 ng/g as laboratory matrix spikes (LMSs). Later, some samples were determined to be above the calibration range and were re-prepared with a single higher LMS at 5000 ng/g and re-analyzed after being appropriately diluted. A stable isotope labeled PFC internal standard r C B-PFOS) and stable isotope labeled surrogate (13C4-PFOS) were added to each sample aliquot prior to solvent extraction, final concentration of each was nominal 1.0 ng/mL in final extract. Samples were extracted by addition of 5.0 mL acetonitrile, briefly mixed and then placed into a freezer for approximately 1 hr to facilitate precipitation of fat and protein solids. Then, samples were centrifuged to remove the solids. A 1.0 mL aliquot of each extract was transferred to an autosampler vial with 0.010 mL formic acid, capped and then analyzed by LC/MS/MS. In some instances, samples were diluted 10fold or 100-fold appropriately with acetonitrile containing IS and SRS at the same levels as previously added to samples during extraction. The first sample preparations began on April 16, 2012 and the last preparations occurred on April 23, 2012. Samples were always prepared as analytical batches, with one analytical batch typically comprised of a set of14 solvent calibration standards ranging from 0.025 ng/mL to 250 ng/mL; two sets of LCSs as described below, several method and solvent blanks, and sample extracts representing approximatelylO study samples (an analytical sample, analytical sample duplicate and LMSs were included per sample). All the analytical batches for this study are listed in Table 6. The typical calibration range for each analytical batch data set was from 0.025 to 100 ng/m L (equivalent 9 Study: IS012-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 to nominal 0.25 to 1000 ng/g of fish tissue concentration), with dropping of the highest prepared calibration standard of 250 ng/mL. In several instances the sample results exceeded the response of the 100 ng/mL calibrant and in those instances the 50 ng/g and 500 ng/g LMS fortification levels were determined to be insufficient relative to the endogenous analyte level because typically the ideal LMS fortifications are between 0.5-times and 10-times the endogenous analyte level per 3M Environmental SOP requirements. In those instances, the samples were re-prepared with a higher LMS fortification at nominal 5000 ng/g concentration with dilutions, as appropriate, and the original sample results ignored and only the re-prepared sample results reported. Exception(s): In one instance in run g120418b, the sample result for IS012-01-01-01-049 sample and its LMSs were above the revised upper limit of quantitation (ULOQ) of 100 ng/mL (i.e. > 1000 ng/g), and a dilution with a 5000 ng/g LMS was accidentally not prepared for that sample. In that instance, the original analysis result along with the high-LMS result (500 ng/g fortification) was reported, despite the fortification level being slightly less than 0.5-times the endogenous level and determined from a high range calibration by inclusion of the 250 ng/mL standard and exclusion of the calibration standards less than 7.5 ng/mL (eq. 75 to 2500 ng/g fish tissue); see analytical run/batch data for g120418b-high range results for that sample's results. This fact was footnoted in the appropriate results tables. Laboratory control spike (LCS) samples were prepared in sets with three levels of nominal 10.0,100 and 1000 ng/g of PFOS and each level prepared in triplicate. Each batch prepared had two sets of LCSs, normal LCSs fortified with L-PFOS the same as calibration standards, and the second set prepared with brPFOS to verify the quantitative accuracy for PFOS typically found in environmental samples which are comprised of a mixture of linear and branched isomers. The LCSs were prepared by fortifying nominal 0.5 gram aliquots of control largemouth bass fillet homogenate (TN08-0193-1/1), purchased from Osage Catfisheries, Inc. (Osage Beach, MO). Equivalent prepared non-fortified method blanks were also prepared with the control bass tissue to establish the endogenous control matrix PFOS level for subtraction from LCS results during LCS recovery calculations. Results of LCSs were used to evaluate the overall method performance during the analyses. Exception(s): In analytical batch m120423a, the typical set of LCSs and method blanks was not prepared and analyzed with the diluted samples analyzed. That run was conducted as a cleanup set for analysis of diluted samples which earlier results were greater than the ULOQ. This analytical batch/run included only equivalently diluted high-level LCSs from the batches those samples were originally prepared and analyzed. Those dilution LCS results are included in the LCS tables and the statistics for the method performance evaluations. 5.4 LC/MS/MS Analysis The analysis of the fish extracts for PFOS was performed by LC/MS/MS as described in method ETS-8-045.1. The relative percent difference (RPD) of the duplicate sample results was a measure of the sample-specific analytical precision. The determined recovery of fortified PFOS from LMS samples was used as a measure of sample-specific analytical accuracy. Recovery of the surrogate ( 3C4-PFOS) from duplicate analytical samples was additionally used as an indirect measure of PFOS recovery and data precision from each sample result. Details of the specific instrument parameters, the liquid chromatography gradient program, and the specific mass transitions analyzed are detailed in the raw data, and are briefly described below in Table 3, Table 4 and Table 5. Additionally, a summary of all analytical runs conducted for collection of the PFOS data reported herein are shown in Table 6. 10 Study: IS012-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 Table 3. In s tru m e n t In fo rm a tio n Instrument Name(s) Analytical Method ID Liquid Chromatograph Guard column Extraction column Analytical column Column Switching Valve Injection Volume Mass Spectrometer Ion Source Polarity Software ETS-MaryAnn (m), ETS-Ginger (g), ETS-Jonas () ETS-8-045.1 Agilent 1100 with Binary Pump Pre-Autosampler; Prism RP (2 x 50 mm, 5 um) Waters Corp. Oasis HLB Online Column (3 x 20 mm, 5 urn); 3 0 ^ Betasil C18 (100 x 2.1 mm, 25 urn); 3 0 ^ 0 to 5 min (waste); 5 minutes to 21 minutes to analytical column 40 nL API 5000 Triple Quadrapole Turbolon Spray (Electrospray) Negative Ion Analyst 1.4.2 Table 4. G rad ie n t L iq u id C h ro m a to g ra p h y C o n d itio n s (ETS-8-045) Step Total Time Number [m in ] 0 0.00 1 3.00 2 3.50 3 13.5 4 15.5 5 16.0 6 18.0 7 18.3 8 21.0 Flow Rate [m U m in] 0.400 0.400 0.400 0.400 0.400 0.400 0.400 0.400 0.400 Percent A [2 mm Aqueous Ammonium Acetate/ 97.0 97.0 70.0 40.0 40.0 10.0 10.0 97.0 97.0 Percent B [ Acetonitrile] 3.0 3.0 30.0 60.0 60.0 90.0 90.0 3.0 3.0 Table 5. M ass T ra n s itio n s Analyte PFOS i3C8-PFOS (IS) Mass Transitions Monitored 499>130, 499>99, 499>80 507>80 i3C4-PFOS (Surrogate) 503>80 N o te : M u ltip le m a s s tra n s itio n s a re su m m e d to d ive the in s tru m e n t re s p o n s e fo r P F O S 11 Study: IS012-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 Table 6. A n a ly tic a l B atches fo r PFOS in Tennessee R iver Fish (A pril 2012) A n a ly tic a l B atch# Analytical Run ID Report Decision Samples Analyzed/Reported (IS01201-01-01-xxxf] 1 j120417a Data Reported -033, -035, -036,-037,-040 2 m120417b Data Reported -022, -025 3 g120418b Data Reported -021, -042, -043, -045, -046, -048, -050 4 g120418b-high range Partial Data Set Reported -049 5 g120420a Data Reported -038, -039, -041,-044, -047, -051, -052, -053 6 m120420a Data Reported -023, -024, -026, -027, -028, -029, -030, -031,-032,-034 7 m120423a Data Reported Select dilutions [a] Samples listed only Indicate which analytical run the reported result for the analytical sample and sample duplicate were determined and may not indicate where the corresponding LMS results were attained since several required dilution and re-analysis in different analytical batches. See the raw data for the identity of the exact analytical batch used for the reported LMS results. 6 Analytical Results All reported concentration results are rounded to three significant figures. Statistical values are rounded to two significant figures. Because of rounding to obtain the limited significant figure values, reported results may vary slightly from those results found in the raw data. 6.1 Calibration Calibration standards were prepared by spiking varying known quantities of L-PFOS and SRS (13C4PFOS), and a fixed quantity of IS (l;,CB-PFOS) into acetonitrile. A set of 14 calibration standards ranging from nominal 0.0250 ng/mL to 250 ng/mL were prepared; equivalent to nominal 0.250 ng/g to 2500 ng/g for fish tissue quantitation. Following analysis of the calibration standards, the nominal concentration of each standard was plotted versus the measured analyte peak area/IS peak area response ratio. A quadratic equation with 1/x weighting was used to fit the calibration response data. Calibration curves were not forced through zero. Calculating the determined standard concentration using the resultant calibration curve and comparing to the actual known concentration confirmed accuracy of each curve point within 100 25% (+ 30% at the LLOQ). The correlation coefficient (r) was greater than 0.999, and coefficient of determination (r2) was greater than 0.990 for all calibration curves. The summary for the calibration standards for each of the analytical data sets used for reporting is shown in Table 7. Exceptions: Typically all calibration standards were analyzed, but the 250 ng/mL standard dropped during construction of the calibration curve. In analytical run g120418b-high range, only the 7.5 ng/mL to 250 ng/mL standards were used to construct the calibration curve to quantitate sample IS012-01-0101-49 within that range. In analytical run m 120423a, the 250 ng/m L standard was not analyzed. 12 Study: IS012-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 Table 7. L-PFOS Solvent Calibration Results A n a ly tic a l Run ID PFOS (ng/m L) STD STD STD STD STD 1 STD 2 STD 3 STD 4 STD 5 STD 6 STD 7 STDS STD 9 STD 14 10 11 12 13 STD CONC. 0.025 0.05 0.1 0.25 0.35 0.5 1 2.5 7.5 10 15 25 100 250 r jl2 0 4 1 7 a 0.0222 0.0459 0.09S9 0.261 0.367 0.509 1.04 2.5S 7.94 9.92 15 24.3 100 excluded 0.9998 m l2 04 17 b 0.0234 0.049 0.0977 0.254 0.354 0.502 1.01 2.6 7.71 9.S5 15.S 23.7 101 excluded 0.9995 jg l2 0 4 1 8 b g l2 0 4 1 8 b high range g l2 0 4 2 0 a 0.0219 excluded 0.0475 0.0999 0.255 0.361 0.515 1.04 2.61 excluded excluded excluded excluded excluded excluded exclude d 7.6 6.43 0.022S 0.04SS 0.101 0.254 0.354 0.505 1.01 2.61 7.62 9.99 9.63 9.93 15.4 16.7 15.4 24.1 27.6 24.2 100 excluded 0.9998 94.S 253 0.9978 100 excluded 0.9998 m l20420a 0.0243 0.05 0.0977 0.247 0.355 0.506 0.999 2.57 7.54 10.1 15.3 24.3 101 excluded 0.9999 m l20423a Average V alue Std Dev. 0.0237 0.04S9 0.101 0.253 0.35 0.496 0.997 2.6S 0.0231 0.0484 0.0994 0.254 0.357 0.506 1.02 2.61 0.00092 0.0014 0.0015 0.0045 0.0061 0.0064 0.019 0.039 7.42 7.47 0.48 10 9.92 0.15 15.7 15.6 0.55 24 24.6 1.34 101 excluded 0.9996 99.7 253 0.9995 2.21 NA 0.00074 RSD Average Recovery n 4.0% 3.0% 1.5% 1.8% 1.7% 1.3% 1.9% 1.5% 6.5% 1.5% 3.5% 5.4% 2.2% NA 0.074% 92.2% 96.7% 99.4% 102% 102% 101% 102% 104% 99.5% 99.2% 104% 98.4% 99.7% 101% NA 6 6 6 6 6 6 6677777 1 7 6.2 Limits of Quantitation (LOQs) The lower limit of quantitation (LLOQ) was the lowest non-zero calibration standard in the curve that met linearity and accuracy requirements (100 + 30%) and for which the area counts were at least 2X that of the average response determined from solvent blanks. The LLOQ for PFOS in this study was typically the nominal concentration 0.0250 ng/mL acetonitrile standards, and correlating to a 0.25 ng/g LLOQ in fish tissues when accounting for the ~10-fold sample dilution which occurs during extraction (nominal 0.5 g tissue diluted with 5 mL acetonitrile). Typically, the 250 ng/mL standard was dropped and the upper limit of quantitation (ULOQ) was revised to the 100 ng/mL calibration standard (i.e. upper limit of nominal 1000 ng/g for PFOS in fish tissues). However, for one run (g120418b) a 6-point high range calibration from 7.5 ng/mL to 250 ng/mL was employed to quantify the results for one sample (ISO-01-01-01-049) which was accidentally not diluted to be captured with a 100 ng/mL ULOQ. That curve met the same calibration curve requirements as defined in section 6.1 and showed good LMS recovery for the high-LMS for that result and was therefore reported. 13 Study: IS012-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 6.3 Continuing Calibration During the course of each analysis, continuing calibration verifications (CCVs) were performed by regular injection of low- to mid-level calibration standards during the analytical run. The back calculated concentration results were used to verify that the instrument response and the initial calibration curve were still in control during the analysis of the entire batch. All CCVs bracketing reported data for PFOS met the method criteria of 100% 25% accuracy and all CCVs for 13C4-PFOS surrogate were within 100 + 25%. The results of CCVs are summarized for each analytical batch in the raw data. Exception(s): In analytical batch/run g120418b, one CCV result for PFOS was 163% recovery due to an irregularly low IS response which was apparently not reflected in the PFOS response for that injection. That CCV immediately followed the analysis of the calibration standards and LCSs in the beginning of the run and only flanked one side of a sample set. The remaining CCVs in that run were all acceptable and therefore the data was reported with a documented method deviation. The reason for the unusual low IS response was not determined. 6.4 Blanks Three types of blanks were analyzed during this analysis: method blanks were prepared with control largemouth bass fillet homogenate (TN08-0193-1/1) and were spiked with IS and surrogate to determine the endogenous PFOS concentration in the control matrix for the LCSs in each analytical batch; (2) a bluegill "sunfish" control matrix (TN11-0194-1/1) was analyzed in analytical runs g120420a and m 120420a to ascertain endogenous PFOS levels in that control matrix, to compare to process control from MSU-WTL which were the same matrix that passed through the homogenization process; and (3) acetonitrile solvent blanks (with and without IS & surrogate) were also included for evaluations. All blank results were carefully evaluated to determine any effects from instrument contamination and analytical carry over and served to aid in defining the LLOQs for quantitation. All of the blank sample results were considered acceptable upon evaluation of the raw data based on the intended use of the different types of blanks. The PFOS results for the bluegill (TN11-0194-1/1) control matrix blank tissue analyses was 0.804 + 0.089 ng/g (n = 6) and that result was used for comparing the homogenization process control sample results. 6.5 Laboratory Control Spikes (LCSs) Laboratory control spikes (LCSs) were prepared by fortifying known quantities of PFOS into 0.5 gram aliquots of a control bluegill fillet tissue. The LCS sets were prepared at three levels (nominal 10, 100 and 1000 ng/g fish tissue), and each level was prepared in triplicate. Two sets of LCSs were prepared for these analyses, one with L-PFOS (same as calibration standards) and the second set fortified with brPFOS (30.6% branched isomer content). The LCS results are used to evaluate the method performance during the study and overall used for calculation of the analytical method uncertainty for the study, as discussed in the conclusion section. The LCS results and the analytical method uncertainties determined from the statistical evaluation of LCS with L-PFOS and with br-PFOS are shown in Table 8 and Table 9, respectively. The method acceptance criteria of 100 30% was met for all LCSs, with a few exceptions as noted in the LCS results tables. The LCS recoveries were calculated using the following equation: (Determined Concentration o f LCS) , LCS Recovery = ------------------------------------------------ - * 100% Spike Concentration 14 Study: IS012-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 Table 8. L-PFOS LCSs and Matrix Blank Results Analytical Run ID ro i--i o(N _Q --1 a(N E _0Q0 "O(NSt Q--01 LCS Sample I.D. 1204 17 -2 -00 2M B 120417-2-003 M B 1204 17 -2 -00 4M B 120417-2-LowLCS 120417-2-LowLCSD 120417-2-LowLCST 12 04 17 -2 -M id LCS 12 04 17 -2 -M id LCSD 12 04 17 -2 -M id LCST 12 04 17 -2 -High LCS 12 04 17 -2 -High LCSD 12 04 17 -2 -High LCST 1204 17 -1 -001M B 1204 17 -1 -00 2M B 120417-1-003 M B 120417-1-LowLCS 120417-1-LowLCSD 120417-1-LowLCST 12 04 17 -1 -M id LCS 12 04 17 -1 -M id LCSD 12 04 17 -1 -M id LCST 12 04 17 -1 -High LCS 12 04 17 -1 -High LCSD 12 04 17 -1 -High LCST 1204 IS-001M B 1204 IS -002 M B 1204 IS -003 M B 1204 lS -L o w LCS 120418-Low LCSD 1 2 0 4 18-Low LCST 120418-Mi dLCS 120418-Mi dLCSD 120418-Mi dLCST 120418-H igh LCS 120418-H igh LCSD .C X t0o0 ^ ro s^ (N 00 o(rNo "oSt 1--1 tu 120418-H igh LCST 120418-Mi dLCS 120418-Mi dLCSD 120418-Mi dLCST 120418-H igh LCS 120418-H igh LCSD 120418-H igh LCST 1204 20 -1 -00 1M B 1204 20 -1 -00 2M B 1204 20 -1 -00 3M B 120420-1-LowLCS 120420-1-LowLCSD 120420-1-LowLCST 12 04 20 -1 -M id LCS M easured PFOS Concentration (ng/g) 0.545 0.576 0.580 12.8 13.1 11.9 108 123 112 >ULOQ >ULOQ >ULOQ 0.590 0.596 0.598 28.9 27.0 25.6 [a] [a] [a] 953 794 901 0.605 0.621 0.555 12.0 11.8 12.8 115 102 107 933 Endogenous PFOS Cone, (ng/g) 0.567 0.595 0.594 900 867 113 100 104 929 913 891 0.643 0.537 0.535 12.2 12.1 10.9 100 0.594 0.572 Fortified Cone, (ng/g) 0.00 10.5 10.9 10.1 102 103 101 994 1060 941 0.00 25.7 26.1 23.0 102 99.5 101 1040 933 1050 0.00 10.4 10.3 11.0 106 96.0 103 1040 1050 1050 106 96.0 103 1040 1050 1050 0.00 10.6 10.1 9.88 95 PFOS Recovery NA 116% 115% 113% 106% 119% 110% NA NA NA NA 110% 101% 109% NA NA NA 91.4% 85.1% 86.0% NA 110% 110% 111% 109% 106% 104% 90.1% 85.8% 82.5% 106% 103% 100% 89.7% 87.0% 84.8% NA 109% 114% 105% 105% Average Recovery NA 114% 111% NA NA 107% NA 87.5% NA 110% 106% 86.1% 103% 87.2% NA 109% 108% Recovery RSD NA 1.5% 6.0% NA NA 4.7% NA 3.9% NA 0.53% 2.4% 4.4% 2.9% 2.8% NA 4.2% 2.8% 15 Study: IS012-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 Table 8. L-PFOS LCSs and Matrix Blank Results Analytical Run ID o("O(tS--NNfr1 ro ("SNfr (N E LCS Sample I.D. M easured PFOS Concentration (ng/g) Endogenous PFOS Cone, (ng/g) 12 04 20 -1 -M id LCS D 12 04 20 -1 -M id LCST 12 04 20 -1 -High LCS 12 04 20 -1 -Hi ghLCSD 12 04 20 -1 -High LCST 12 04 20 -1 -High LCS DF:100 12 04 20 -1 -Hi ghLCSD DF:100 12 04 20 -1 -High LCST DF:100 1204 20 -1 -00 1M B 1204 20 -1 -00 2M B 1204 20 -1 -00 3M B 120420-1-LowLCS 120420-1-LowLCSD 120420-1-LowLCST 12 04 20 -1 -M id LCS 12 04 20 -1 -M id LCS D 12 04 20 -1 -M id LCST 12 04 20 -1 -High LCS 12 04 20 -1 -Hi ghLCSD 12 04 20 -1 -High LCST 12 04 20 -1 -High LCS DF:100 12 04 20 -1 -Hi ghLCSD DF:100 12 04 20 -1 -High LCST DF:100 12 04 20 -1 -High LCS df:10 12 04 20 -1 -Hi ghLCSD df:10 12 04 20 -1 -High LCST df:10 12 04 17 -2 -High LCS df:10 12 04 17 -2 -Hi ghLCSD df:10 12 04 17 -2 -High LCST df:10 120 114 1090 >ULOQ >ULOQ 10.1 10.1 10.0 0.643 0.545 0.5 OS 12.3 11.9 10.7 96.6 117 111 721 S16 960 s.ss 9.25 10.0 94S 953 1030 1050 934 1040 0.565 0.565 [bl 0 .5 7 1 [bl A verag e R e co ve ry (Low LCS) Stan d ard D eviation (Low LCS) RSD (Low LCS) n A v e ra g e R e co v e ry (M id LCS) S ta n d a rd D e v ia tio n (M id LCS) RSD (M id LCS) n A verag e R e co ve ry (H igh LCS) S tand ard D eviation (H igh LCS) RSD (H igh LCS) n A v erag e R e co ve ry (H igh LCS-D iluted) S tand ard D eviation (H igh LCS-D iluted) RSD (High LCS-D iluted) n Fortified Cone, (ng/g) 108 106 929 916 950 929 916 950 0.00 10.6 10.1 9.SS 95 107.6 106 929 916 950 929 916 950 929 916 950 1040 933 1050 PFOS Recovery 111% 108% 117% NA NA 108% 110% 105% NA 110% 112% 103% 101% 108% 105% 77.6% 89.1% 101% 95.5% 101% 105% 102% 104% 108% 101% 100% 99.0% Average Recovery 117% 108% NA 108% 104% 89.2% 101% 105% 99.9% 110% 4.2% 3.8% 15 107% 4.5% 4.2% 15 89.8% 10% 11% 13 103% 4.4% 4.2% 12 Recovery RSD NA 2.4% NA 4.4% 3.4% 13% 4.9% 2.9% 0.82% 16 Study: IS012-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 Table 8. L-PFOS LCSs and Matrix Blank Results Analytical Run ID ampe - - M easured PFOS Concentration (ng/g) Endogenous PFOS Cone, (ng/g) Fortified Cone, (ng/g) PFOS Recovery Average Recovery Recovery RSD A v e ra g e R e co v e ry (All Levels LCSs) 103% S tan d ard D e via tio n (All Levels LCSs) 9.7% RSD (All Levels LCSs) 9.4% n 55 2 x S td . Dev. (A nalytical M e th o d U n ce rta in ty fo r L-PFOS) 19% Note: The e n d o g en o u s level is th e average re su lt fro m th re e an alyze d m atrix bla nks fo r th e bass c o n tro l tissu e T N O S-0193 used fo r LCS preparations. NA; not applicable [a] IS ac cid e n tally w as n o t ad ded to th e p re p a ra tio n and LCS v alu e not d e te rm in e d [b] Endogenous level w as n o t d e te rm in e d in th e a n alytica l run, so p re vio u s re s u lt w a s used (w hen m o re th a n o n e re su lt w a s available, p re fe re n ce w as given to the endogenous PFOS result obtained on the same instrum ent) Table 9. brPFOS LCSs and Matrix Blank Results Analytical Run ID ro i--i a(N _Q i--1 "Sfr (N E _Q 0i--01 a(i--QN01 LCS Sample I.D. 1204 17 -2 -00 2M B 120417-2-003 M B 1204 17 -2 -00 4M B 12 04 17 -2 -BLLow LCS 120417-2-BLLowLCSD 12 04 17 -2 -BLLow LCST 12 04 17 -2 -B LM id LCS 1 2 0 4 1 7 -2 -B LM id LC S D 12 04 17 -2 -B LM id LCST 1 2 0 4 1 7 -2 - B LH ighLCS 1 2 0 4 1 7 -2 -B LH ighLCSD 1 2 0 4 1 7 -2 - B LH ighLC ST 1204 17 -1 -001M B 1204 17 -1 -00 2M B 120417-1-003 M B 12 04 17 -1 -BLLow LCS 120417-1-BLLowLCSD 12 04 17 -1 -BLLow LCST 12 04 17 -1 -B LM id LCS 1 2 0 4 1 7 -1 -B LM id LC S D 12 04 17 -1 -B LM id LCST 1 2 0 4 1 7 -1 - B LH ighLCS 1 2 0 4 1 7 -1 -B LH ighLCSD 1 2 0 4 1 7 -1 - B LH ighLC ST 1204 IS-001M B 1204 IS -002 M B 1204 IS -003 M B 1204 lS -B L L o w LCS 12041S-BLLOW LCSD 1204 lS -B L L o w LCST M easured PFOS Concentration (ng/g) 0.545 0.576 0.5S0 12.6 10.6 11.4 112 113 122 >ULOQ >ULOQ >ULOQ 0.54S 0.549 0.615 12.7 12.5 11.9 119 103 116 >ULOQ >ULOQ >ULOQ 0.605 0.621 0.555 12. S 12.2 11.8 Endogenous PFOS Cone, (ng/g) Fortified Cone, (ng/g) PFOS Recovery Average Recovery Recovery RSD 0.567 0.595 0.594 0.00 10.3 9.29 9.64 95.6 92.0 97.2 1000 934 1020 0.00 10.5 10.8 9.97 104 92.7 105 1020 999 975 0.00 10.7 10.6 10.5 NA 117% 108% 113% 117% 122% 125% NA NA NA NA 115% 110% 114% 114% 111% 110% NA NA NA NA 114% 110% 107% NA 113% 121% NA NA 113% 112% NA NA 110% NA 4.1% 3.4% NA NA 2.4% 1.9% NA NA 3.2% 17 Study: IS012-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 Table 9. brPFOS LCSs and Matrix Blank Results Analytical Run ID .c X - Cl ni S ^S (N d ro (N O (N & ro (N a (N E ro (N a (i--N1 E Measured PF05 LCS Sample I.D. Concentration 1 2 0 4 1 8 -B L M id LC S (ng/g) 125 1 2 0 4 1 8 -B L M id LC S D 107 1 2 0 4 1 8 -B L M id LC S T 1 2 0 4 18-BLHigh LCS 108 >ULOQ 120418-BLHigh LCSD >ULOQ 1 2 0 4 18-BLHigh LCST >ULOQ 1 2 0 4 1 8 -B L M id LC S 126 1 2 0 4 1 8 -B L M id LC S D 107 1 2 0 4 1 8 -B L M id LC S T 108 1 2 0 4 18-BLHigh LCS 1140 1 2 0 4 18-BLHigh LCSD 1300 1 2 0 4 18-BLHigh LCST 1250 1204 20 -1 -00 1M B 0.643 1204 20 -1 -00 2M B 1204 20 -1 -00 3M B 0.537 0.535 12 04 20 -1 -BLLow LCS 11.5 12 04 20 -1 -BLLow LCSD 10.3 12 04 20 -1 -BLLow LCST 11.4 12 04 20 -1 -B LM id LCS 113 12 04 20 -1 -B LM id LCSD 113 12 04 20 -1 -B LM id LCST 117 1 2 0 4 2 0 -1 - B LH ighLCS >ULOQ 1 2 0 4 2 0 -1 -B LH ighLCSD >ULOQ 1 2 0 4 2 0 -1 - B LH ighLC ST >ULOQ 1204 20 -1 -00 1M B 0.643 1204 20 -1 -00 2M B 0.545 1204 20 -1 -00 3M B 0.508 12 04 20 -1 -BLLow LCS 11.1 12 04 20 -1 -BLLow LCSD 9.98 12 04 20 -1 -BLLow LCST 11.0 12 04 20 -1 -B LM id LCS 111 12 04 20 -1 -B LM id LCSD 12 04 20 -1 -B LM id LCST 110 114 1 2 0 4 2 0 -1 - B LH ighLCS >ULOQ 1 2 0 4 2 0 -1 -B LH ighLCSD >ULOQ 1 2 0 4 2 0 -1 - B LH ighLC ST >ULOQ 120420-1-BLHighLCS df:10 1005 120420-1-BLHighLCSD df:10 996 120420-1-BLHighLCST df:10 1030 120417-2-BLHighLCS df:10 1110 120417-2-BLHighLCSD df:10 1050 120417-2-BLHighLCST df:10 979 A verag e R ecovery (Low LCS) Endogenous PFOS Cone, (ng/g) 0.594 0.572 0.565 0.565 [bl 0 .5 7 1 [bl Standard D e via tion (Low LCS) RSD (Low LCS) n Average Recovery (M id LCS) Standard D eviation (M id LCS) Fortified Cone, (ng/g) 109 95.8 94.9 981 1030 1010 109 95.8 94.9 981 1030 1010 0.00 9.76 9.19 9.95 104 93.5 110 948 958 976 0.00 9.76 9.19 9.95 104 93.5 110 948 958 976 948 958 976 1020 999 975 PFOS Recovery 114% 111% 113% NA NA NA 98.5% 94.4% 96.5% 115% 125% 123% NA 112% 107% 109% 108% 120% 106% NA NA NA NA 108% 103% 105% 106% 117% 103% NA NA NA 106% 104% 105% 109% 105% 100% Average Recovery 113% NA 96.5% 121% NA 109% 111% NA NA 105% 109% NA 105% 105% 110% 4.0% 3.6% 15 113% 6.3% Recovery RSD 1.4% NA 2.1% 4.4% NA 2.3% 6.9% NA NA 2.4% 6.9% NA 1.0% 4.1% 18 Study: IS012-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 Table 9. brPFOS LCSs and Matrix Blank Results Analytical Run ID LCS Sample I.D. M easured PFOS Concentration (ng/g) RSD (M id LCS) Endogenous PFOS Cone, (ng/g) Fortified Cone, (ng/g) PFOS Recovery Average Recovery 5.6% Recovery RSD n 15 A verag e Recovery (High LCS) NA Standard D eviation (High LCS) NA RSD (High LCS) NA n0 A verage Recovery (High LCS-Diluted) 105% Standard D eviation (High LCS) 2.8% RSD (High LCS-Diluted) 2.7% n6 Average Recovery (All Levels LCSs) 110% Standard Deviation (All Levels LCSs) 5.7% RSD (All Levels LCSs) 5.1% n 36 2 x Std. Dev. (Analytical M eth o d U ncertainty for L-PFOS) 11% Note: The e n d o g en o u s level is th e average re su lt fro m th re e an alyze d m atrix bla nks fo r th e bass c o n tro l tissu e T N O S-0193 used fo r LCS preparations. NA; not applicable [a] IS ac cid e n tally n o t ad ded to th e p re p a ra tio n and LCS valu e not d e te rm in e d [b] Endogenous level w as n o t d e te rm in e d in th e a n alytica l run, so p re vio u s re s u lt w a s used (w hen m o re th a n o n e re su lt w a s available, p re fe re n ce w as given to the endogenous PFOS result obtained on the same instrum ent) 6.6 Laboratory Matrix Spikes (LMSs) Laboratory matrix spikes (LMSs) were generated by fortifying known quantity of L-PFOS to an aliquot of fish fillet tissue and then preparing the sample the same as regular non-fortified samples. For this study, each sample had at least two LMSs prepared, and several had a third prepared at a higher level following initial analyses. The original two LMSs were fortified at nominal 50 and 500 ng/g, and for approximately Vi of the samples either the 50 ng/g or 500 ng/g LMS was sufficient relative to the endogenous level to evaluate the sam pie-specific PFOS recovery. However, for the other Vi of sam pies the levels were significantly higher and required a LMS fortified at 5000 ng/g to sufficiently evaluate recovery. The most appropriate LMS fortification for evaluating recovery is typically between 0.5-times and 10-times the endogenous level determined for the non-fortified sample. Exception(s): For the three homogenization process control fish tissues, the nominal 50 ng/g LMS was used to evaluate PFOS recovery, but was ~ 50-times the endogenous PFOS level in the corresponding non-fortified samples. Additionally, for sample ISO-12-01-01-049, the 500 ng/g LMS result was used to evaluate PFOS recovery, but was slightly less than 0.5-times the endogenous level of the corresponding non-fortified sample result. A SOP deviation was included in the raw data to accommodate these exceptions. Overall, the LMS recoveries for L-PFOS fortified into the fish samples ranged from 76.7% to 125%, with an overall mean recovery + standard deviation of 98.9% + 10% (n=29 Tennessee river fish fillet homogenates). The LMS recoveries for the pre-, mid- and post-study process controls were 102%, 99.1% and 102%, respectively. The LMS recovery from the analysis of the NIST SRM-1947 Lake 19 Study: IS012-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 Michigan fish tissue homogenate was 95.3%. The method acceptance criteria of 100 30% was met for all LMS results. The LMS recovery results are summarized in Table 1 and the individual sample results used for calculating LMS recoveries are shown in TablelO. The calculation of PFOS recoveries used the following equation: T,, ,, (Determined Concentration of LMS - Determined Concentration o f Field Sample) LMS Recovery = - ---------------------------------------------------------------------------------------------------------- ---- *100% Spike Concentration 6.7 NIST SRM 1947 Results The MSU-WTL was provided two aliquots of a Lake Michigan trout standard reference material (SRM 1947) purchased from the National Institutes of Standards and Technology (NIST). The reference value of 5.9 + 0.39 ng/g PFOS was recently published for NIST SRM 1947 [Reiner et. al. 2012. Anal Bioanal Chenr, DOI 10.1007/s00216-012-5943-5], Two aliquots of NIST SRM 1947 were provided to the MSU-WTL prior to the commencement of the study. One non-processed aliquot was subsequently returned to the 3M Environmental Laboratory with the processed Tennessee River fish samples and process controls for analysis. The second aliquot of NIST SRM 1947 (not-processed) was sent to Axys Laboratories by MSU-WTL to be analyzed with the three field duplicates. The 3M Environmental Laboratory's determined average + RPD value for the NIST SRM 1947 sample was 6.71 ng/g + 0.27 ng/g PFOS for duplicate analytical sample results. The nominal 50 ng/g LMS recovery for the SRM 1947 was 95.3%. The results show that the average PFOS concentration determ ined for SRM 1947 was accurate to 114% with respect to the N 1ST SRM 1947 reference value. 6.8 Process Control Results The MSU-WTL was provided three ~25 g aliquots of 3M-hom ogenized control bluegill fillet tissue TN110194-1/1 that was purchased from Osage Catfisheries, Inc. (Osage Beach, Mo) prior to the study. One aliquot was planned to be homogenized at MSU prior to the homogenization of Tenn. River Fish (pre study process control), one near the middle of the process of homogenizing the Tenn. River fish (mid study process control), and one after the homogenization of the Tenn. R. fish was completed (post-study process control). Those homogenization process controls were returned with the sample shipments for analysis of PFOS and compared to the PFOS results of for the same control bluegill tissue, but which did not travel to MSU-WTL for processing. The results of those comparisons are shown in Table 10. The results showed that the RPD was less than 15% for each and that no significant contamination occurred during the homogenization process and shipment process that would have affected the PFOS results of this study. Table 10. Homogenization Process Control Results Sample ID Sample Description PFOS (ng/g) TN 11-0194-1/1 Bluegill fillet control tissue TN 11-0194-1/1 0.804 ISO12-01-01-01-021 Pre-Study Process Control (TNI 1-0194-1/1) 0.S15 ISO12-01-01-01-022 Mid-Study Process Control (TN 11-0194-1/1) 0.909 ISO12-01-01-01-042 Post-Study Process Control (TNI 1-0194-1/1) 0.912 Relative percent difference (RPD) w as calculated for the control tissue result relative to th e process control result. RPD NA 1.3% 12% 13% 20 Study: IS012-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 6.9 Individual Sample Results The individual analytical sample results are shown in Table 11. A summary of the average concentrations with their RPDs and LMS recoveries determined from these results are provided in T a b le ! Table 1 ! Individual PFOS Results for Fillet Tissues from Fish Collected in the Tennessee River, April 2012 LIM S S a m p le ID Sam ple D escription 1SO12-01-01-01-0 21 1SO12-01-01-01-0 21 Dup 1SO12-01-01-01-0 21 LMS 1SO12-01-01-01-0 22 1SO12-01-01-01-022 Dup 1SO12-01-01-01-022 LMS 1SO12-01-01-01-0 23 1SO12-01-01-01-023 Dup 1SO12-01-01-01-023 LMS 1SO12-01-01-01-0 24 1SO12-01-01-01-024 Dup 1SO12-01-01-01-024 LMS 1SO12-01-01-01-0 25 1SO12-01-01-01-025 Dup 1SO12-01-01-01-025 LMS 1SO12-01-01-01-0 25 1SO12-01-01-01-025 Dup 1SO12-01-01-01-025 LMS 1SO12-01-01-01-0 27 1SO12-01-01-01-027 Dup 1SO12-01-01-01-027 LMS 1SO12-01-01-01-0 28 1SO12-01-01-01-028 Dup 1SO12-01-01-01-028 LMS 1SO12-01-01-01-0 29 1SO12-01-01-01-029 Dup 1SO12-01-01-01-029 LMS 1SO12-01-01-01-0 30 1SO12-01-01-01-030 Dup 1SO12-01-01-01-030 LMS 1SO12-01-01-01-0 31 1SO12-01-01-01-0 31 Dup 1SO12-01-01-01-0 31 LMS 1SO12-01-01-01-0 32 1SO12-01-01-01-032 Dup 1SO12-01-01-01-032 LMS 1SO12-01-01-01-0 33 1SO12-01-01-01-033 Dup Pre-Study, Process Control #1 (TN11-0194-1/1) Pre-Study, Process Control #1 Dup (TNI 1-0194-1/1) Pre-Study, Process Control #1 LMS (TNI 1-0194-1/1) Mid-Study, Process Control #2 (TNI 1-0194-1/1) Mid-Study, Process Control #2 Dup (TN11-0194-1/1) Mid-Study, Process Control #2 LMS (TN11-0194-1/1) MSF 01120403 MSF 01120403 Dup MSF 01120403 LMS High MSF 02 120403 MSF 02 120403 Dup MSF 02 120403 LMS MSF 03 120403 MSF 03 120403 Dup MSF 03 120403 LMS MSF 04 120403 MSF 04 120403 Dup MSF 04 120403 LMS MSF 05 120403 MSF 05 120403 Dup MSF 05 120403 LMS MSF 06 120403 MSF 06 120403 Dup MSF 06 120403 LMS MSF 07 120403 MSF 07 120403 Dup MSF 07 120403 LMS MSF 08 120403 MSF 08 120403 Dup MSF 08 120403 LMS MSF 09 120403 MSF 09 120403 Dup MSF 09 120403 LMS MSF 10 120403 MSF 10 120403 Dup MSF 10 120403 LMS MSF 21120404 MSF 21120404 Dup PFOS Cone. (ng/g) 0.806 0.823 52.2 1.07 0.752 53.2 4810 4740 9340 4120 4350 8580 71.4 70.0 108 595 558 1150 460 483 955 152 160 581 2900 2950 8140 1350 1360 6270 2070 1970 7480 1740 1930 7170 651 692 A v g PFOS C o n c e n tra tio n (ng/g) R P D (%) Surrogate (CrPFOS) Recovery w LM S Spike C o n c e n tra tio n (ng/g) 0.815 52.2 0.909 [bl 53.2 4770 9340 4240 8580 70.7 108 576 1150 471 955 156 581 2920 8140 1360 6270 2020 7480 1840 7170 671 2.0% NA 34% NA 1.4% NA 5.3% NA 1.9% NA 6.5% NA 4.9% NA 5.4% NA 1.6% NA 0.13% NA 5.2% NA 10% NA 6.1% 100% 100% 102% 105% 105% 107% 98.6% 107% 101% 107% 103% 98.4% 101% 102% 101% 100% 102% 98.9% 102% 100% 100% 103% 105% 107% 103% 101% 101% 101% 104% 100% 104% 106% 100% 94.8% 91.0% 100% 89.7% 96.3% NA 50.5 [dl NA 52.8 [dl NA 4730 NA 4700 NA 46.0 NA 524 NA 482 NA 554 NA 5280 NA 4920 NA 5280 NA 5060 NA LM S Spike Recovery (% ) NA 102% NA 99.1% NA 96.5% NA 92.4% NA 81.0% NA 108% NA 100% NA 76.7% NA 98.7% NA 99.8% NA 104% NA 105% NA 21 Study: IS012-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 Table 11. Individual PFOS Results for Fillet Tissues from Fish Collected in the Tennessee River, April 2012 LIM S S a m p le ID Sam ple D escription ISO12-01-01-01-033 LMS 1SO12-01-01-01-0 34 1SO12-01-01-01-034 Dup 1SO12-01-01-01-034 LMS 1SO12-01-01-01-0 35 1SO12-01-01-01-035 Dup 1SO12-01-01-01-035 LMS 1SO12-01-01-01-0 35 1SO12-01-01-01-035 Dup 1SO12-01-01-01-035 LMS 1SO12-01-01-01-0 37 1SO12-01-01-01-037 Dup 1SO12-01-01-01-037 LMS 1SO12-01-01-01-0 38 1SO12-01-01-01-038 Dup 1SO12-01-01-01-038 LMS 1SO12-01-01-01-0 39 1SO12-01-01-01-039 Dup 1SO12-01-01-01-039 LMS 1SO12-01-01-01-040 Dup 1SO12-01-01-01-040 1SO12-01-01-01-040 LMS 1SO12-01-01-01-041 1SO12-01-01-01-041 Dup 1SO12-01-01-01-041 LMS 1SO12-01-01-01-042 1SO12-01-01-01-042 Dup 1SO12-01-01-01-042 LMS 1SO12-01-01-01-043 1SO12-01-01-01-043 Dup 1SO12-01-01-01-043 LMS 1SO12-01-01-01-044 1SO12-01-01-01-044 Dup 1SO12-01-01-01-044 LMS 1SO12-01-01-01-045 1SO12-01-01-01-045 Dup 1SO12-01-01-01-045 LMS 1SO12-01-01-01-045 1SO12-01-01-01-045 Dup 1SO12-01-01-01-045 LMS 1SO12-01-01-01-047 1SO12-01-01-01-047 Dup 1SO12-01-01-01-047 LMS 1SO12-01-01-01-048 1SO12-01-01-01-048 Dup 1SO12-01-01-01-048 LMS 1SO12-01-01-01-049 1SO12-01-01-01-049 Dup 1SO12-01-01-01-049 LMS 1SO12-01-01-01-0 50 MSF 21120404 LMS MSF 22 120404 MSF 22 120404 Dup MSF 22 120404 LMS MSF 23 120404 MSF 23 120404 Dup MSF 23 120404 LMS MSF 24 120405 MSF 24 120405 Dup MSF 24 120405 LMS MSF 25 120405 MSF 25 120405 Dup MSF 25 120405 LMS MSF 26 120405 MSF 26 120405 Dup MSF 26 120405 LMS MSF 27 120403 MSF 27 120403 Dup MSF 27 120403 LMS MSF 28 120404 Dup MSF 28 120404 MSF 28 120404 LMS MSF 29 120405 MSF 29 120405 Dup MSF 29 120405 LMS Post Study, Process Control #3 Post Study, Process Control #3 Dup (TN11-0194-1/1) Post Study, Process Control #3 LMS (TNI 1-0194-1/1) Reference Tissue (NIST SRM Reference Tissue Dup (NIST Reference Tissue 1/2 LMS MSF 1112-04-03 MSF 1112-04-03 Dup MSF 1112-04-03 LMS MSF 12 12-04-03 MSF 12 12-04-03 Dup MSF 12 12-04-03 LMS MSF 13 12-04-03 MSF 13 12-04-03 Dup MSF 13 12-04-03 LMS MSF 14 12-04-03 MSF 14 12-04-03 Dup MSF 14 12-04-03 LMS MSF 15 12-04-03 MSF 15 12-04-03 Dup MSF 15 12-04-03 LMS MSF 16 12-04-04 MSF 16 12-04-04 Dup MSF 16 12-04-04 LMS MSF 17 12-04-04 PFOS Conc. (ng/g) 1080 4570 4740 9650 52.4 52.5 552 464 409 884 312 300 999 2100 2080 6830 2060 2180 7380 172 188 783 2050 2050 7100 0.984 0.839 Avg. PFOS Con cent rat ion (ng/g) 1080 4660 9650 52.5 552 437 884 306 999 2090 6830 2120 7380 180 783 2050 7100 0.912 R P D (%) Surrogate (CrPFOS) Recovery w NA 3.5% NA 0.0056% NA 12% NA 3.8% NA 1.0% NA 5.4% NA 9.2% NA 0.063% NA 16% 90.1% 90.7% 94.1% 98.3% 97.8% 102% 91.9% 102% 87.1% 98.8% 95.8% 101% 91.0% 94.0% 100% 100% 95.1% 96.1% 102% 98.2% 107.9% 97.1% 92.0% 94.4% 102% 101% 99.0% LM S Spike C o n c e n tra tio n (ng/g) 478 NA 5010 NA 502 NA 494 NA 554 NA 5030 NA 4950 NA 487 NA 4953 NA 56.3 6.85 6.58 54.9 2990 2880 7580 537 515 5450 942 958 5810 3360 3370 8270 718 790 5670 1550 151 1500 w 1950 w 165 56.3 6.71 54.9 2940 7580 526 5450 950 5810 3370 8270 754 5670 1520 1950 165 NA 4.0% NA 3.9% NA 4.2% NA 1.7% NA 0.505 NA 9.6% NA 3.2% NA 0.75% 103% 98.8% 102% 100% 100% 101% 101% 103% 99.0% 104% 96.5% 102% 100% 100% 97.6% 100% 98.7% 102% 102% 100% 99.0% 100% 101% 5 4 .1 [dl NA 50.6 NA 4970 NA 4880 NA 4910 NA 4810 NA 4940 NA 493 w NA LM S Spike Recovery (% ) 86.0% NA 99.6% NA 99.7% NA 90.7% NA 125% NA 94.2% NA 106% NA 124% NA 102% NA 102% NA 95.3% NA 93.5% NA 101% NA 99.0% NA 102% NA 99.4% NA 86.9% NA 22 Study: IS012-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 Table 11. Individual PFOS Results for Fillet Tissues from Fish Collected in the Tennessee River, April 2012 LIM S S a m p le ID Sam ple D escription PFOS Conc. (ng/g) Avg. PFOS C o n c e n tra tio n (ng/g) R P D (%) Surrogate (CrPFOS) Recovery w LM S Spike C o n c e n tra tio n (ng/g) LM S Spike R e c o v e ry (%) 15012-01-01-01-050 Dup 15012-01-01-01-050 LMS 15012-01-01-01-051 15012-01-01-01-051 Dup 15012-01-01-01-051 LMS 15012-01-01-01-052 15012-01-01-01-052 Dup 15012-01-01-01-052 LMS 15012-01-01-01-053 15012-01-01-01-053 Dup 15012-01-01-01-053 LMS MSF 17 12-04-04 Dup MSF 17 12-04-04 LMS MSF 1812-04-04 MSF 1812-04-04 Dup MSF 1812-04-04 LMS MSF 19 12-04-04 MSF 19 12-04-04 Dup MSF 19 12-04-04 LMS MSF 2012-04-04 MSF 2012-04-04 Dup MSF 2012-04-04 LMS 166 581 710 691 6080 672 678 5620 1170 1180 6150 581 701 6080 675 5620 1180 6150 NA 2.7% NA 0.86% NA 0.82% NA 100% 100% 100% 101% 99.0% 101% 102% 100% 101% 100% 101% 493 NA 5220 NA 4780 NA 4780 84.2% NA 103% NA 104% NA 104% A ll concentration values are rounded to 3 significant figures for reporting. Statistical results are rounded to 2 significant figures. As a result, raw data values may vary slightly from those reported NA; not applicable [ajSurrogate was spiked into sam pie aliquots at nom inal 1.00 ng/g, result is shown for the least diluted sample result [b] RPD >20% [c] The LMS spike concentration was less than K of the endogenous PFOS concentration for the non-fortified sample results [d] The LMS spike concentration was greater than 10-times of the endogenous PFOS concentration for the non-fortified sample results [e] Value was determined from a six point high-range calibration curve from 7.5 ng/mL to 250 ng/m Lfor quantifying PFOS in tissues from nominal 75.0 to 2500 ng/g. All other reported results were obtained with calibration from 0.025 ng/mL to 100 ng/mL and the 250 ng/mL standard excluded, and samples were typically diluted to within that range when they exceeded 1000 ng/g. 7 Conclusion The determination of perfluorooctane sulfonate (PFOS) concentrations for 29 largemouth bass fillet tissues (skin removed) collected from the Tennessee River in April 2012 near Decatur, Alabama was successfully completed. In addition, 3 homogenization process controls and one NIST SRM 1947 standard reference material were analyzed. This analysis was conducted in support of the Weston Solutions, Inc. project plan, titled "Work Plan for Fish Sampling and Analysis for PFOS in Largemouth Bass from the Tennessee River in the Vicinity of the 3M Decatur Facility, Decatur, Alabam a". The analytical aspects reported herein were conducted in the 3M Environmental Laboratory as 3M study IS012-01-01-01. Fish fillets (skin removed) were prepared in the field by Weston Solutions, Inc. personnel after being collected from the Tennessee River and the shipped to the Michigan State University Wildlife Toxicology Laboratory where fish fillet homogenates were prepared. The frozen homogenates were then shipped to the 3M Environmental Laboratory in two shipments that were received on April 14, 2012 and April 17, 2012. Quantitative analysis for PFOS was performed from April 16, 2012 through April 24, 2012 following analytical method ETS-8-045.1. The determined average concentrations of PFOS in the Tennessee River fish tissue samples ranged from 52.5 ng/g to 4770 ng/g. The recoveries of fortified PFOS from the LMS samples prepared for each Tennessee River fish tissue samples were within 100 +25% recovery, ranging from 76.7% to 125%, with an overall average + standard deviation of 98.9% + 10%. Additionally, the average recovery and standard deviation for a surrogate recovery standard (T3C4-PFOS) for all of the Tennessee River fish samples was 99.5% + 4.3%. The determined average PFOS value and relative difference for the NIST SRM 1947 fish tissue that was also received was 6.71 + 0.27 ng/g (4.0% RPD); a 114% recovery relative to the NIST reference value of 5.90 + 0.39 ng/g. 23 Study; IS 0 12-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 The results for the three process controls showed that contamination during homogenization and shipping from MSU-WTL to the 3M Environmental Laboratory did not occur at any significant level that would affect the results. The analytical method uncertainty for the data set was determined based on LCS results. Sets of LCSs were prepared with each extraction batch by spiking L-PFOS or br-PFOS into control fish tissue homogenates at nominal 10, 100 and 1000 ng/g, each level in triplicate. The method uncertainty is the range about the mean with which it is expected to statistically find the reported results with 95 out o f 100 measurements, referred to as the 95% confidence interval {95% Cl), and is + 2 Std. Deviations of the LCS data. For L-PFOS the method uncertainty (95% Cl) was + 19% and for brPFOS (30.6% branched isomers) the method uncertainty (95% Cl) was + 11%. 8 Data/Sample Retention All fish samples and associated project data (hardcopy and electronic) will be archived according to 3M Environmental Laboratory standard operating procedures as project ISO12-01 -01-01. 9 List of Attachments Attachment A: Preparation Forms, Raw Data and Chromatograms 10 Signatures Report Approval: Cleston C. Lange, Ph.D., Principal Analytical Investigator ^ Z 20>Z Date William K. Reagen, Ph.D., 3M Environmental Laboratory Management c o j Date QAU Repres^enittaatthive 24 5 - 2 - IX Date Study: IS012-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 Attachment A Preparation Forms, Raw Data and Chromatograms (Available Upon Request) 25 3M Environmental Laboratory Report No IS012-01-01-02 Final Report Analysis of Water Samples from the Tennessee R iv e r-A p ril 2012 Laboratory Request Number: IS012-01-01-02 Method Requirement: 3M Method ETS-8-044.1 Report Date - April 26, 2012 Testing Laboratory 3M Environmental Health and Safety Operations Environmental Laboratory 3M Center, Bldg 260-05-N-17 St. Paul, MN 55144 Requester Gary Hohenstein 3M EHS Operations 3M Building 224-5W-03 Saint Paul, MN 55144-1000 Phone: (651) 737-3570 Testing Cert #2052.01 The testing reported herein meet the requirem ents o f ANSI/ ISO/IEC 17025:2005 "General Requirem ents fo r the Com petence o f Testing and Calibration Laboratories" , in accordance with the A 2LA Testing Certificate #2052.01. Testing that com plies with this International Standard also m eets p rincip le s o f ISO 9001:2000. PAGE 1 OF 11 3M Environmental Laboratory Report No IS012-01-01-02 3M Environmental Laboratory 3M Environmental Laboratory Manager: William K. Reagen, Ph.D. 3M Principal Analytical Investigator and Report Author: Susan W olf A nalytical R ep o rt IS 0 1 2-01 -01 -02 Analysis of Water Samples from the Tennessee River - April 2012 Report Date: April 26, 2012 1 S um m ary/lntroduction The 3M Environmental Laboratory prepared and analyzed samples collected by Weston personnel from the Tennessee River, conducted per the study work plan titled 'W ork Plan for Fish Sampling and Analysis for PFOS in Largemouth Bass from the Tennessee River in the Vicinity of the 3M Decatur Facility". Samples were collected April 5 and 6, 2012. Samples were returned to the 3M Environmental Laboratory on April 13, 2012 at ambient conditions and analyzed for perfluorooctane sulfonate (PFOS) under 3M Environmental Laboratory project number IS012-01-01-02. The 3M Environmental Laboratory prepared sample containers for nine sampling locations, however only three sampling locations were collected. Each empty container was marked with a "fill to here'' line that corresponded to a final volume of 200 mL. Each sample set consisted of a field sample, field sample duplicate, and afield matrix spike. All samples bottles included the addition of internal standard [i3Cei]-PFOS and surrogate recovery standard [13C4]-PFOS, which were added to the sample containers prior to being sent to the field for sample collection. Sample bottles reserved for field matrix spikes were fortified with appropriate matrix spike solutions containing PFOS, internal standard [l;,CB]-PFOS and surrogate recovery standard [l3C4]-PFOS prior to being sent to the field for sample collection. Samples were prepared and analyzed using either direct injection or solvent dilution according to 3M Environmental Laboratory method ETS-8-044.1 "Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis". The internal standard [i 3Cb]-PFOS was used to aid in the quantitation of the samples. Table 1 summarizes the sample results using the analytical method identified above. All results for quality control samples prepared and analyzed with the samples will be reported and discussed elsewhere in this report. PAGE 2 OF 11 3M Environmental Laboratory Report No 5012-01-01-02 Table 1. Sample Results Summary. 3M LIM S ID IS 0 12-01-01-02-001 IS 0 1 2 -0 1 -0 1 -0 2 -0 0 2 IS 0 12-01-01-02-004 IS 0 1 2 -0 1 -0 1 -0 2 -0 0 5 IS 0 1 2 -0 1 -0 1 -0 2 -0 0 7 IS 0 1 2 -0 1 -0 1 -0 2 -0 0 8 IS 0 1 2 -0 1 -0 1 -0 2 -0 1 0 IS 012-01-01-02-011 IS 0 1 2 -0 1 -0 1 -0 2 -0 1 2 IS 0 1 2 -0 1 -0 1 -0 2 -0 1 3 Sam ple D escription PFOS C oncentration (ng/m L) Location 1 - Sample Location 1 - Sample Duplicate A v e ra g e % R P D S am p le/S am p le Dup Location 2 - Sample Location 2 - Sample Duplicate A v e ra g e % R P D S am p le/S am p le Dup Location 3 - Sample Location 3 - Sample Duplicate A v e ra g e % R P D S am p le/S am p le Dup Equipm ent Blank 1 Equipm ent Blank 2 Equipm ent Blank 3 Trip Blank 0.516 0.450 0 .4 8 3 14 0.720 0.661 0 .6 9 1 8 .5 0.0210 0.0263 0 .0 2 3 7 2 2 121 <0.00928 <0.00928 <0.00928 <0.00928 NA = Not Applicable (1 ) T he analytical m ethod uncertainty fo rth e reported results using m ethod ETS-8-044.1 by direct Inject fo rP F O S Is 18%. (2) T he sample / sample duplicate R PD value did not m eet method acceptance criteria of <20% . 2 M ethod Sum m ary 2.1 Methods Analysis for PFOS was completed following 3M Environmental Laboratory method ETS-8-044.1 "Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS". Table 2. Target Analytes. Target Analytes Perfluorooctanesulfonate (C8 Sulfonate) Acronym PFOS Reference Material Structure Linear + Branched 2.2 Sample Collection Water samples were collected in 250 mL NalgeneTM (high-density polyethylene) bottles prepared at the 3M Environmental Laboratory. A laboratory prepared Trip Blank and Trip Blank field matrix spike were sent with the set of collection bottles. Sample bottles were received by the laboratory at ambient conditions on April 13, 2012. Samples were stored refrigerated at the laboratory after receipt. 2.3 Sample Preparation An aliquot of sample was removed for analysis and placed in an autovial. During the preparation of the laboratory control samples, an aliquot of a separate internal standard spiking solution and surrogate recovery standard solution was added to the laboratory control samples (nominal concentration of 0.1 ng/mL). The sample bottles were spiked with an internal standard at a PAGE 3 OF 11 3M Environmental Laboratory Report No 5012-01-01-02 nominal concentration of 0.1 ng/mL and a surrogate recovery standard at a nominal concentration of 0.025 ng/ml prior to being sent to the field for sample collection. 2.4 Analysis All samples and quality control samples were analyzed for PFOS and the surrogate recovery standard [13C4]-PFOS using high performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS). Detailed instrument parameters, the liquid chromatography gradient program, and the specific mass transitions analyzed are described in the raw data hard copies placed in the final data packet, and are briefly described below. Table 3. Instrument Parameters. Instrument Name Analysis Dates Analytical Method Liquid Chromatograph Guard column Analytical column Injection Volume Mass Spectrometer Ion Source Electrode Polarity Software ETS Kirk 4/24/12 ETS-8-044.1 Aqilent 1290 Betasil C18 (4.6 mmX 100 mm), 5 ix Betasil C18 (4.6 mmX 100 mm), 5|x 50 uL Applied Biosystems API 5500 Turbo Spray Turbo ion electrode Negative Analyst 1.5.2 Table 4. Liquid Chromatography Conditions. Step Number 0 1 2 3 4 5 6 7 8 9 Total Time (min) 0.00 0.50 4.00 6.00 11.0 13.0 13.5 16.0 16.5 19.0 Flow Rate (fJJmin) 750 750 750 750 750 750 750 750 750 750 Percent A (5mMammonium acetate:0.1% acetic acid) 90.0 90.0 70.0 70.0 20.0 20.0 10.0 10.0 90.0 90.0 Percent B (Methanol) 10.0 10.0 30.0 30.0 80.0 80.0 90.0 90.0 10.0 10.0 PAGE 4 OF 11 3M Environmental Laboratory Report No 5012-01-01-02 Table 5. Mass Transitions. Analyte Mass Tiansition Q1/Q3 Internal Standard Mass Tiansition Q1/Q3 499/99 PFOS 499m t 3Cs]-PFOS 507m 499/130 [13CJ-PFOS 503m f 3CaJ-PFOS 507m Dwell time was 30 msec for each transition. The individual transitions were summed to produce a "total ion chromatogram" (TIC), which was used for quantitation. 3 A n alytical R esults 3.1 Calibration Samples were analyzed for PFOS and [13C4]-PFOS against a stable isotope internal standard calibration curve. Calibration standards were prepared by spiking known amounts of the stock solution containing the target analyte and surrogate recovery standard into a laboratory-prepared synthetic groundwater containing calcium and magnesium. A separate internal standard spiking solution was prepared and an aliquot was added at the same level to all calibration standards and laboratory control samples at a nominal concentration of 1 ng/mL. A calibration curve ranging from approximately 0.005 ng/mL to 15 ng/mL was analyzed (0.005 ng/mL to 5 ng/mL for [13C4]-PFOS). The low and high curve points were disabled for PFOS to meet accuracy or method blank criteria. A quadratic, 1/x weighted, calibration curve of the standard peak area ratios was used to fit the data for each analyte. The data were not forced through zero during the fitting process. Calculating the standard concentrations using the peak area ratios and the resultant calibration curve confirmed accuracy erf each curve point. Each curve point was quantitated using the overall calibration curve and reviewed for accuracy. Method calibration accuracy requirements of 10025% (10030% for the lowest curve point) were met for all analytes. The correlation coefficient (r) was greater than 0.995 for PFOS and [13C4]-PFOS. It should be noted that the calibration curve that was used to analyze the samples on 4/24/12, expired on 4/17/12. The expiration date assigned to the curve was based on the expiration date of the solutions containing the surrogate recovery standard [13C4]-PFOS, which were used to prepared the calibration standards. The solutions containing PFOS that were used to prepare the calibation curve were not expired. The solutions containing f 3C4]-PFOS that were used to prepared the high set of LCS samples and the sample collection bottles, were not expired. The high set of LCSs for [13C4]-PFOS surrogate met method acceptance criteria, as did the [13C4]-PFOS surrogate in the Tennessee River samples. A method delation is included in the raw data. 3.2 System Suitability A calibration standard was analyzed four times at the beginning of the analytical sequence to demonstrate overall system suitability. The acceptance criteria of less than or equal to 5% relative standard deviation (RSD) for peak area ratio and retention time criteria of less than or equal to 2% RSD were met for PFOS and [13C4]-PFOS. 3.3 Limit of Quantitation (LOQ) The LOQ for this analysis is the lowest non-zero calibration standard in the curve that meets linearity and accuracy requirements and for which the area counts are at least twice those of the appropriate blanks. The LOQ for PFOS was 0.00928 ng/m L. PAGE 5 OF 11 3M Environmental Laboratory Report No 5012-01-01-02 3.4 Continuing Calibration During the course of each analytical sequence, continuing calibration verification samples (CCVs) were analyzed to confirm that the instrument response and the initial calibration curve were still in control. All reported sample results were bracketed by CCVs that met method criteria of 100% 25%. 3.5 Blanks Three types of blanks were prepared and analyzed with the samples: solvent blanks (procedural blanks), field/trip blanks, and equipment rinseate blanks for the groundwater samples. Each blank result was reviewed and used to evaluate method performance. Procedural blank results were reviewed according to the method and used to evaluate method performance to determine the LOQ for each analyte. 3.6 Lab Control Spikes (LCSs) Low, mid, and high lab control spikes were prepared and analyzed in triplicate for PFOS, while only low and high lab control spikes were prepared for the [13C4]-PFOS surrogate. The LCS samples were prepared by spiking known amounts of the analyte into 10 mL of synthetic groundwater to produce the desired concentration. The spiked water samples (LCSs) were then analyzed in the same manner as the samples. The method acceptance criteria states that the average recovery of LCSs be 100% 20% with a RSD <20%, when evaluated independently at each concentration level. All LCSs met these criteria. The LCS results were used to determine overall method uncertainty in Section 3.7. The following calculations were used to generate data in Table 6 for laboratory control spikes. LCS Percent Recovery C alculated C oncentration ^ g o / Spike C oncentration standard deviation LCS replicates LCS% RSD = 100% average LCS recovery PAGE 6 OR 11 Table 6. Laboratory Control Spike Recovery. ETS-8-044.1 Analyzed 4/24/12 Lab ID LC S-120424-1 L C S -1 20424-2 L C S -1 20424-3 Average % R S D L C S -1 20424-4 L C S -1 20424-5 L C S -1 20424-6 Average % R S D L C S -1 20424-7 L C S -1 20424-8 L C S -1 20424-9 Average % R S D PFOS (Linear + Branched) S p ik e d C oncentration (ng/m L) C alculated C oncentration (ng/m L) % Recovery 0.0231 0.0231 0.0231 0.185 0.185 0.185 4.64 4.64 4.64 0.0273 0.0257 0.0253 113% 3 .9 % 0.174 0.186 0.175 9 6 .1% 3 .6 % 3.75 4.01 4.19 8 5 .8% 5.7 % 118 111 110 93.8 100 94.4 80.7 86.4 90.4 ETS-8-044.1 Analyzed 4/24/12 Lab ID L C S -1 20424-1 (1) L C S -1 2 0 4 2 4 -2 (1) L C S -1 2 0 4 2 4 -3 (1) Average % R S D L C S -1 20424-4 L C S -1 20424-5 L C S -1 20424-6 Average % R S D [ I3C 4] - P F 0 S S p ik e d C oncentration (ng/m L) C alculated C oncentration (ng/m L) 0.0238 0.0238 0.0238 0.190 0.190 0.190 0.0246 0.0249 0.0240 1 0 3% 1.9% 0.204 0.200 0.202 106% 0 .9 4 % % Recovery 103 105 101 107 105 106 (1) The solution used to prepare this set of LCS samples expired 4/17/12. 3M Environmental Laboratory Report No 5012-01-01-02 PAGE 7 OF 11 3M Environmental Laboratory Report No 5012-01-01-02 3.7 Analytical Method Uncertainty Analytical uncertainty is based on historical QC data that is control charted and used to evaluate method accuracy and precision. The method uncertainty is calculated following ETS-12-012.2. The standard deviation is calculated for the set of accuracy results (in %) obtained for the QC samples. For method ETS-8-044.1, the most recent fifty QC samples were used. The analytical method uncertainty is calculated by multiplying the standard deviation by a factor of 2, which corresponds to a confidence level of 95%. Table 7. Analytical Method Uncertainty A n a ly te PFOS [13C 4 ]-P F 0 S Standard D eviation 8.75 6.74 M ethod U ncertainty (% ) 18 13 3.8 Field Matrix Spikes (FMS) Afield matrix spike was collected at each sampling location to verify that the analytical method is applicable to the collected matrix. Field matrix spikes are generated by adding a measured volume of field sample to a container spiked by the laboratory with the target analytes prior to shipping sample containers for sample collection. Field matrix spike recoveries within method acceptance criteria of 10030% confirm that "unknown" components in the sample matrix do not significantly interfere with the extraction and analysis of the analytes of interest. The reference standards for PFOS in the field matrix spiking solution consisted of linear and branched isom ers. In addition, field matrix spikes for this project also included the use of stable isotope surrogate spikes of [13C4]-PFOS, which was added at a nominal concentration of 0.025 ng/mL to all sample bottles prior to sample collection. Surrogate matrix spike recoveries within method acceptance criteria of 10030% confirm that "unknown" components in the sample matrix do not significantly interfere with the preparation and analysis of the analytes of interest. Table 8. Field Matrix Spike Level. S am p lin g lo cation All locations and Trip Blank P FO S ng /m L 0.0232 FMS Recovery - (amP*e Concentration of FM S- Average Concentration :Field Sample & FieldSampleDup.) *jqq0/ Spike Concentration 4 Data Summary and Discussion The tables below summarize the sample results and field matrix spike and surrogate recoveries for the sampling locations as well as the Trip Blanks. Results and average values are rounded to three significant figures according to EPA rounding rules. Because of rounding, values may vary slightly from those listed in the raw data. Field matrix spike and surrogate recoveries meeting the method acceptance criteria of 30%, demonstrate that the method(s) were appropriate for the given matrix and their respective quantitative ranges. PAGE 8 OF 11 3M Environmental Laboratory Report No 5012-01-01-02 Table 9. Tennessee River; Location 1 3 M U M S ID D es crip tio n IS 0 1 2 -0 1 -0 1 -0 2 -0 0 1 Location 1 IS 0 1 2 -0 1 -0 1 -0 2 -0 0 2 Location 1 Sam ple Duplicate IS 0 1 2 -0 1 -0 1 -0 2 -0 0 3 Location 1 FMS A verage Concentration (ng/mL) % R P D /% R S D PROS Concentration (ng/m L) % R eco very 0.516 NA 0.450 NA 0.537 NC 0.483 ng/m L 14% NA = Not Applicable NC = Not Calculated; E ndogenous sam ple concentration Is g re a te rth a n 2k spiking level. C3c 4i-p f o s % R ecovery 108 105 106 106% 0.99% Table 10. Tennessee River; Location 2 3 M U M S ID D es crip tio n IS 0 12-01-01-02-004 Location 2 IS 0 1 2 -0 1 -0 1 -0 2 -0 0 5 Location 2 Sam ple Duplicate IS 0 1 2 -0 1 -0 1 -0 2 -0 0 6 Location 2 FMS A verage Concentration (ng/mL) % R P D /% R S D PROS Concentration (ng/m L) % R ecovery 0.720 NA 0.661 NA 0.654 NC 0.691 n g/m Lt 8.5% NA = Not Applicable NC = Not Calculated; E ndogenous sam ple concentration Is g re a te rth a n 2k spiking level. [ 13C4]-PROS % R ecovery 108 106 109 108% 1.5% Table 11. Tennessee River; Location 3 3 M U M 5 ID D es crip tio n IS 0 1 2 -0 1 -0 1 -0 2 -0 0 7 Location 3 IS 0 1 2 -0 1 -0 1 -0 2 -0 0 8 Location 3 Sam ple Duplicate IS 0 1 2 -0 1 -0 1 -0 2 -0 0 9 Location 3 FMS A verage Concentration (ng/mL) % R P D /% R S D PROS Concentration (ng/m L) % R ecovery 0.0210 NA 0.0263 NA 0.0430 83.4 0.0237 ng/rnL 2 2% (V NA = Not Applicable (1 ) T he sample / sam ple duplicate R P D did not m eet m ethod acceptance criteria o f <20% . f 13C4]-PROS % R ecovery 105 108 106 106% 1.6% PAGE 9 OF 11 Table 12. Rinseate Blanks 3MUMSID IS 0 1 2 -0 1 -0 1 -0 2 -0 1 0 IS 0 1 2 -0 1 -0 1 -0 2 -0 1 1 IS 0 1 2 -0 1 -0 1 -0 2 -0 1 2 Description Equipm ent Blank 1 Equipm ent Blank 2 Equipm ent Blank 3 Table 13. Trip Blanks 3MUMSID IS 0 1 2 -0 1 -0 1 -0 2 -0 1 3 IS 0 1 2 -0 1 -0 1 -0 2 -0 1 4 Description T rip Blank T rip Blank FMS 3M Environmental Laboratory Report No 5012-01-01-02 PFOS Concentration (ng/mL) <0.00928 <0.00928 <0.00928 f n C4]-PFOS %Recovery 108 105 108 PFOS Concentration (ng/mL) %Recovery <0.00928 0.0209 NA 90.1 [ 13C4]-PFOS VoRecwerv 105 103 PAGE 10 OF 11 3M Environmental Laboratory Report No. IS012-01-01-02 5 Conclusion Laboratory control spikes were used to determine the analytical method accuracy and precision for PFOS. The accuracy and precision were then used to estimate the method uncertainty for the results. Field matrix spike recoveries demonstrated that the analytical method was appropriate for the given sample matrix. Analysis was completed using 3M Environmental Laboratory method ETS-8-044.1 "Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis". Analytical results are reported in Tables 1 and 9-13 of this report. 6 Data/Sam ple Retention All remaining samples and associated project data (hardcopy and electronic) will be archived according to 3M Environmental Laboratory standard operating procedures. William K. Reagen, Ph.D,, 3M Environmental Laboratory Manager Date The 3M Environmental Laboratory's Quality Assurance Unit has audited the data and report for this project. Quality Assurance Representative PAGE 11 OF 11 APPENDIX B AXYS LABORATORY ANALYTICAL DATA PACKAGE www.axysanalytical.com AXYS Axys Analytical Services Ltd 2045 Mills Road West SIDNEY, BRITISH COLUMBIA, CANADA V8L 5X2 TEL 250-655-5800 FAX250-655-5811 w w w .axysanalytical.com AXYS Client No.: Client Address: 4673 Cardno ENTRIX 4295 Okemos Road, Suite 101 Okemos, MI, US, 48864 The AXYS contact for these data is Kalai Pillay. Page 1 of 49 www.axysanalytical.com BATCH SUMMARY Batch ID: WG39768 A na lysis Type: Perfluorinated Organic C ontract: Sam ples: L17838-1 L17838-2 L17838-3 L17838-4 BATCH MAKEUP 4673 Reference Tiss. TCR11-0039 MSF05D 120403 MSF11D 120403 MSF24D 120405 Date: 27-Apr-2012 M a trix Type: Tissue B lank: WG39768-101 Reference o r S pike: WG39768-102 W G 39768-104 W G 39768-105 D uplicate: WG39768-103 Com m ents: 1. Data are not blank corrected. FQA-006 Rev. 2. 18-Jul-1994 Copyright AXYS Analytical Services Ltd February 1993 Page 2 of 49 www.axysanalytical.com AXYS METHOD MLA-043 Rev 8 Form 1A PERFLUORINATED ORGANICS ANALYSIS REPORT AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811 Contract No.: 4673 Project No. Lab Sample I.D.: CLIENT SAMPLE NO. Reference Tiss. TCR11-0039 Sample Collection: N/A 3M - ALABAM A L17838-1 Matrix: TISSUE Sample Size: 2.00 g (wet) Sample Receipt Date: 19 - A p r - 2 0 1 2 Initial Calibration Date: 24-Apr-2012 Extraction Date: 23-Apr-2012 Instrument ID: LC MS/MS Analysis Date: 25-Apr-2012 Time: 00:32:37 Column ID: C18MS Extract Volume (uL): 4000 Sample Data Filename: FC2G_157 S: 24 Injection Volume (uL): 15 Blank Data Filename: F C 2 G _ 1 57 S: 22 Dilution Factor: N/A Cal. Ver. Data Filename: F C 2 G 157 S: 15 Concentration Units: ng/g (wet weight basis) This page is part of a total report that contains information necessary for accreditation compliance. Results are compliant with NELAP accreditation described in the total report. Sample results relate only to the sample tested. COMPOUND LAB FLAG 1 CONC. FOUND REPORTING LIMIT (RL)2 RETENTION TIME PFOS 7.16 1.00 (L) 8:17 (1) W here applicable, custom lab flags have been used on this report. (2) Reporting Limit (Code): S = sam ple detection limit; M = method detection limit; L = lowest calibration level equivalent; Q = contract defined limit. T hese data are validated and reported a s accurate and in accord with A X Y S Analytical S e rv ice s Ltd. ISO 17025 com pliant quality assu ran ce processes. S igned :____________ B r y a n A l o n z o _________ For Axys Internal Use Only [ XSL Template: FC-Form1A.xsl; Created: 27-Apr-2012 14:59:50; Application: XMLTransformer-1.12.13; Report Filename: PFC_FC_LC_PFOA_L17838-1_Form1A_FC2G_157S24_SJ1453688.html; Workgroup: WG39768; Design ID: 1722 ] Page 3 of 49 Page 1 of 1 (WG39768 -PFC_FC_LC_PFOA_L17838-1_Form1A_FC2G_157S24_SJ1453688.html) www.axysanalytical.com AXYS METHOD MLA-043 Rev 8 Form 2 PERFLUORINATED ORGANICS ANALYSIS REPORT AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811 Contract No.: 4673 Project No. Lab Sample I.D.: CLIENT SAMPLE NO. Reference Tiss. TCR11-0039 Sample Collection: N/A 3M - ALABAM A L17838-1 Matrix: TISSUE Sample Size: 2.00 g (wet) Sample Receipt Date: 19-Apr-2012 Initial Calibration Date: 24-Apr-2012 Extraction Date: 23-Apr-2012 Instrument ID: LC MS/MS Analysis Date: 25-Apr-2012 Time: 00:32:37 Column ID: C18MS Extract Volume (uL): 4000 Sample Data Filename: FC2G_157 S: 24 Injection Volume (uL): 15 Blank Data Filename: F C 2 G _ 1 57 S: 22 Dilution Factor: N/A Cal. Ver. Data Filename: F C 2 G 157 S: 15 Concentration Units: ng absolute This page is part of a total report that contains information necessary for accreditation compliance. Results are compliant with NELAP accreditation described in the total report. Sample results relate only to the sample tested. LABELED COMPOUND LAB FLAG1 SPIKE CONC. CONC. FOUND R(%) 2 RETENTION TIME 13C4-PFOS 36.0 31.5 87.6 8:17 CLEANUP STANDARD 13C8-PFOA 12.0 10.6 88.5 7:04 (1) W here applicable, custom lab flags have been used on this report. (2) R(%) = percent recovery. T hese data are validated and reported a s accurate and in accord with A X Y S Analytical S e rv ice s Ltd. ISO 17025 com pliant quality assu ran ce processes. S igned :____________ B r y a n A l o n z o _________ For Axys Internal Use Only [ XSL Template: FC-Form2.xsl; Created: 27-Apr-2012 14:59:50; Application: XMLTransformer-1.12.13; ReportFilename: PFC_FC_LC_PFOA_L17838-1_Form2_FC2G_157S24_SJ1453688.html; Workgroup: WG39768; Design ID: 1722 ] Page 1 of 1 (WG39768 -PFC_FC_LC_PFOA_L17838-1_Form2_FC2G_157S24_SJ1453688.html) Page 4 of 49 www.axysanalytical.com AXYS METHOD MLA-043 Rev 8 Form 1A PERFLUORINATED ORGANICS ANALYSIS REPORT AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811 Contract No.: 4673 Project No. Lab Sample I.D.: CLIENT SAMPLE NO. MSF05D 120403 Sample Collection: 03-Apr-2012 3M - ALABAM A L17838-2 (A) Matrix: TISSUE Sample Size: 2.11 g (wet) Sample Receipt Date: 19 - A p r - 2 0 1 2 Initial Calibration Date: 24-Apr-2012 Extraction Date: 23-Apr-2012 Instrument ID: LC MS/MS Analysis Date: 25-Apr-2012 Time: 0 1 :39:33 Column ID: C18MS Extract Volume (uL): 4000 Sample Data Filename: FC2G_157 S: 28 Injection Volume (uL): 15 Blank Data Filename: F C 2 G _ 1 57 S: 22 Dilution Factor: N/A Cal. Ver. Data Filename: F C 2 G 157 S: 15 Concentration Units: ng/g (wet weight basis) This page is part of a total report that contains information necessary for accreditation compliance. Results are compliant with NELAP accreditation described in the total report. Sample results relate only to the sample tested. COMPOUND LAB FLAG 1 CONC. FOUND REPORTING LIMIT (RL)2 RETENTION TIME PFOS 656 0.948(L) 8:17 (1) W here applicable, custom lab flags have been used on this report. (2) Reporting Limit (Code): S = sam ple detection limit; M = method detection limit; L = lowest calibration level equivalent; Q = contract defined limit. T hese data are validated and reported a s accurate and in accord with A X Y S Analytical S e rv ice s Ltd. ISO 17025 com pliant quality assu ran ce processes. S igned :____________ B r y a n A l o n z o _________ For Axys Internal Use Only [ XSL Template: FC-Form1A.xsl; Created: 27-Apr-2012 14:59:50; Application: XMLTransformer-1.12.13; Report Filename: PFC_FC_LC_PFOA_L17838-2_Form1A_FC2G_157S28_SJ1453692.html; Workgroup: WG39768; Design ID: 1722 ] Page 5 of 49 Page 1 of 1 (WG39768 -PFC_FC_LC_PFOA_L17838-2_Form1A_FC2G_157S28_SJ1453692.html) www.axysanalytical.com AXYS METHOD MLA-043 Rev 8 Form 2 PERFLUORINATED ORGANICS ANALYSIS REPORT AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811 Contract No.: 4673 Project No. Lab Sample I.D.: CLIENT SAMPLE NO. MSF05D 120403 Sample Collection: 03-Apr-2012 3M - ALABAM A L17838-2 (A) Matrix: TISSUE Sample Size: 2.11 g (wet) Sample Receipt Date: 19-Apr-2012 Initial Calibration Date: 24-Apr-2012 Extraction Date: 23-Apr-2012 Instrument ID: LC MS/MS Analysis Date: 25-Apr-2012 Time: 0 1 :39:33 Column ID: C18MS Extract Volume (uL): 4000 Sample Data Filename: FC2G_157 S: 28 Injection Volume (uL): 15 Blank Data Filename: F C 2 G _ 1 57 S: 22 Dilution Factor: N/A Cal. Ver. Data Filename: F C 2 G 157 S: 15 Concentration Units: ng absolute This page is part of a total report that contains information necessary for accreditation compliance. Results are compliant with NELAP accreditation described in the total report. Sample results relate only to the sample tested. LABELED COMPOUND LAB FLAG1 SPIKE CONC. CONC. FOUND R(%) 2 RETENTION TIME 13C4-PFOS 36.0 28.4 79.0 8:17 CLEANUP STANDARD 13C8-PFOA 12.0 9.77 81.5 7:06 (1) W here applicable, custom lab flags have been used on this report. (2) R(%) = percent recovery. T hese data are validated and reported a s accurate and in accord with A X Y S Analytical S e rv ice s Ltd. ISO 17025 com pliant quality assu ran ce processes. S igned :____________ B r y a n A l o n z o _________ For Axys Internal Use Only [ XSL Template: FC-Form2.xsl; Created: 27-Apr-2012 14:59:50; Application: XMLTransformer-1.12.13; ReportFilename: PFC_FC_LC_PFOA_L17838-2_Form2_FC2G_157S28_SJ1453692.html; Workgroup: WG39768; Design ID: 1722 ] Page 1 of 1 (WG39768 - PFC_FC_LC_PFOA_L17838-2_Form2_FC2G_157S28_SJ1453692.html) Page 6 of 49 www.axysanalytical.com AXYS METHOD MLA-043 Rev 8 Form 1A PERFLUORINATED ORGANICS ANALYSIS REPORT AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811 Contract No.: 4673 Project No. Lab Sample I.D.: CLIENT SAMPLE NO. MSF11D 120403 Sample Collection: 03-Apr-2012 3M - ALABAM A L17838-3 N (A) Matrix: TISSUE Sample Size: 2.15 g (wet) Sample Receipt Date: 19 - A p r - 2 0 1 2 Initial Calibration Date: 24-Apr-2012 Extraction Date: 23-Apr-2012 Instrument ID: LC MS/MS Analysis Date: 25-Apr-2012 Time: 16:11:38 Column ID: C18MS Extract Volume (uL): 4000 Sample Data Filename: FC2G_158 S: 8 Injection Volume (uL): 15 Blank Data Filename: F C 2 G _ 1 57 S: 22 Dilution Factor: 5 Cal. Ver. Data Filename: F C 2 G 158 S: 4 Concentration Units: ng/g (wet weight basis) This page is part of a total report that contains information necessary for accreditation compliance. Results are compliant with NELAP accreditation described in the total report. Sample results relate only to the sample tested. COMPOUND LAB FLAG 1 CONC. FOUND REPORTING LIMIT (RL)2 RETENTION TIME PFOS D 3720 4.65 (L) 8:21 (1) W here applicable, custom lab flags have been used on this report; D = dilution data. (2) Reporting Limit (Code): S = sam ple detection limit; M = method detection limit; L = lowest calibration level equivalent; Q = contract defined limit. T hese data are validated and reported a s accurate and in accord with A X Y S Analytical S e rv ice s Ltd. ISO 17025 com pliant quality assu ran ce processes. S igned :____________ B r y a n A l o n z o _________ For Axys Internal Use Only [ XSL Template: FC-Form1A.xsl; Created: 27-Apr-2012 14:59:50; Application: XMLTransformer-1.12.13; ReportFilename: PFC_FC_LC_PFOA_L17838-3_Form1A_FC2G_158S8_SJ1453777.html; Workgroup: WG39768; Design ID: 1722 ] Page 1 of 1 (WG39768 -PFC_FC_LC_PFOA_L17838-3_Form1A_FC2G_158S8_SJ1453777.html) Page 7 of 49 www.axysanalytical.com AXYS METHOD MLA-043 Rev 8 Form 2 PERFLUORINATED ORGANICS ANALYSIS REPORT AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811 Contract No.: 4673 Project No. Lab Sample I.D.: CLIENT SAMPLE NO. MSF11D 120403 Sample Collection: 03-Apr-2012 3M - ALABAM A L17838-3 N (A) Matrix: TISSUE Sample Size: 2.15 g (wet) Sample Receipt Date: 19-Apr-2012 Initial Calibration Date: 24-Apr-2012 Extraction Date: 23-Apr-2012 Instrument ID: LC MS/MS Analysis Date: 25-Apr-2012 Time: 16:11:38 Column ID: C18MS Extract Volume (uL): 4000 Sample Data Filename: FC2G_158 S: 8 Injection Volume (uL): 15 Blank Data Filename: F C 2 G _ 1 57 S: 22 Dilution Factor: 5 Cal. Ver. Data Filename: F C 2 G 158 S: 4 Concentration Units: ng absolute This page is part of a total report that contains information necessary for accreditation compliance. Results are compliant with NELAP accreditation described in the total report. Sample results relate only to the sample tested. LABELED COMPOUND LAB FLAG1 SPIKE CONC. CONC. FOUND R(%) 2 RETENTION TIME 13C4-PFOS D 36.0 20.4 56.5 8:21 CLEANUP STANDARD 13C8-PFOA D 12.0 8.90 74.2 7:06 (1) W here applicable, custom lab flags have been used on this report; D = dilution data. (2) R(%) = percent recovery. T hese data are validated and reported a s accurate and in accord with A X Y S Analytical S e rv ice s Ltd. ISO 17025 com pliant quality assu ran ce processes. S igned :____________ B r y a n A l o n z o _________ For Axys Internal Use Only [ XSL Template: FC-Form2.xsl; Created: 27-Apr-2012 14:59:50; Application: XMLTransformer-1.12.13; ReportFilename: PFC_FC_LC_PFOA_L17838-3_Form2_FC2G_158S8_SJ1453777.html; Workgroup: WG39768; Design ID: 1722 ] Page 1 of 1 (WG39768 -PFC_FC_LC_PFOA_L17838-3_Form2_FC2G_158S8_SJ1453777.html) Page 8 of 49 www.axysanalytical.com AXYS METHOD MLA-043 Rev 8 Form 1A PERFLUORINATED ORGANICS ANALYSIS REPORT AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811 Contract No.: 4673 Project No. Lab Sample I.D.: CLIENT SAMPLE NO. MSF24D 120405 Sample Collection: 05-Apr-2012 3M - ALABAM A L17838-4 Matrix: TISSUE Sample Size: 2.05 g (wet) Sample Receipt Date: 19 - A p r - 2 0 1 2 Initial Calibration Date: 24-Apr-2012 Extraction Date: 23-Apr-2012 Instrument ID: LC MS/MS Analysis Date: 25-Apr-2012 Time: 0 1 :22:47 Column ID: C18MS Extract Volume (uL): 4000 Sample Data Filename: FC2G_157 S: 27 Injection Volume (uL): 15 Blank Data Filename: F C 2 G _ 1 57 S: 22 Dilution Factor: N/A Cal. Ver. Data Filename: F C 2 G 157 S: 15 Concentration Units: ng/g (wet weight basis) This page is part of a total report that contains information necessary for accreditation compliance. Results are compliant with NELAP accreditation described in the total report. Sample results relate only to the sample tested. COMPOUND LAB FLAG 1 CONC. FOUND REPORTING LIMIT (RL)2 RETENTION TIME PFOS 537 0.976 (L) 8:17 (1) W here applicable, custom lab flags have been used on this report. (2) Reporting Limit (Code): S = sam ple detection limit; M = method detection limit; L = lowest calibration level equivalent; Q = contract defined limit. T hese data are validated and reported a s accurate and in accord with A X Y S Analytical S e rv ice s Ltd. ISO 17025 com pliant quality assu ran ce processes. S igned :____________ B r y a n A l o n z o _________ For Axys Internal Use Only [ XSL Template: FC-Form1A.xsl; Created: 27-Apr-2012 14:59:50; Application: XMLTransformer-1.12.13; ReportFilename: PFC_FC_LC_PFOA_L 17838-4_Form1 A_FC2G_157S27_SJ1453691.html; Workgroup: W G39768;DesignID: 1722 ] Page 9 of 49 Page 1 of 1 (WG39768 -PFC_FC_LC_PFOA_L17838-4_Form1A_FC2G_157S27_SJ1453691.html) www.axysanalytical.com AXYS METHOD MLA-043 Rev 8 Form 2 PERFLUORINATED ORGANICS ANALYSIS REPORT AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811 Contract No.: 4673 Project No. Lab Sample I.D.: CLIENT SAMPLE NO. MSF24D 120405 Sample Collection: 05-Apr-2012 3M - ALABAM A L17838-4 Matrix: TISSUE Sample Size: 2.05 g (wet) Sample Receipt Date: 19-Apr-2012 Initial Calibration Date: 24-Apr-2012 Extraction Date: 23-Apr-2012 Instrument ID: LC MS/MS Analysis Date: 25-Apr-2012 Time: 0 1 :22:47 Column ID: C18MS Extract Volume (uL): 4000 Sample Data Filename: FC2G_157 S: 27 Injection Volume (uL): 15 Blank Data Filename: F C 2 G _ 1 57 S: 22 Dilution Factor: N/A Cal. Ver. Data Filename: F C 2 G 157 S: 15 Concentration Units: ng absolute This page is part of a total report that contains information necessary for accreditation compliance. Results are compliant with NELAP accreditation described in the total report. Sample results relate only to the sample tested. LABELED COMPOUND LAB FLAG1 SPIKE CONC. CONC. FOUND R(%) 2 RETENTION TIME 13C4-PFOS 36.0 27.3 75.9 8:17 CLEANUP STANDARD 13C8-PFOA 12.0 10.8 90.3 7:06 (1) W here applicable, custom lab flags have been used on this report. (2) R(%) = percent recovery. T hese data are validated and reported a s accurate and in accord with A X Y S Analytical S e rv ice s Ltd. ISO 17025 com pliant quality assu ran ce processes. S igned :____________ B r y a n A l o n z o _________ For Axys Internal Use Only [ XSL Template: FC-Form2.xsl; Created: 27-Apr-2012 14:59:50; Application: XMLTransformer-1.12.13; ReportFilename: PFC_FC_LC_PFOA_L17838-4_Form2_FC2G_157S27_SJ1453691.html; Workgroup: WG39768; Design ID: 1722 ] Page 10 of 49 Page 1 of 1 (WG39768 - PFC_FC_LC_PFOA_L17838-4_Form2_FC2G_157S27_SJ1453691.html) www.axysanalytical.com AXYS METHOD MLA-043 Rev 8 Form 1A PERFLUORINATED ORGANICS ANALYSIS REPORT AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811 Contract No.: 4673 Project No. Lab Sample I.D.: CLIENT SAMPLE NO. Lab Blank Sample Collection: N/A N/A W G 39768-101 Matrix: TISSUE Sample Size: 2.00 g Sample Receipt Date: N/A Initial Calibration Date: 24-Apr-2012 Extraction Date: 23-Apr-2012 Instrument ID: LC MS/MS Analysis Date: 24-Apr-2012 Time: 23:59:06 Column ID: C18MS Extract Volume (uL): 4000 Sample Data Filename: FC2G_157 S: 22 Injection Volume (uL): 15 Blank Data Filename: F C 2 G _ 1 57 S: 22 Dilution Factor: N/A Cal. Ver. Data Filename: F C 2 G _ 1 57 S: 15 Concentration Units: ng/g This page is part of a total report that contains information necessary for accreditation compliance. Results are compliant with NELAP accreditation described in the total report. Sample results relate only to the sample tested. COMPOUND LAB FLAG 1 CONC. FOUND REPORTING LIMIT (RL)2 RETENTION TIME PFOS U 1.00 (L) (1) W here applicable, custom lab flags have been used on this report; U = not detected at RL. (2) Reporting Limit (Code): S = sam ple detection limit; M = method detection limit; L = lowest calibration level equivalent; Q = contract defined limit. T hese data are validated and reported a s accurate and in accord with A X Y S Analytical S e rv ice s Ltd. ISO 17025 com pliant quality assu ran ce processes. S igned :____________ B r y a n A l o n z o _________ For Axys Internal Use Only [ XSL Template: FC-Form1A.xsl; Created: 27-Apr-2012 14:59:50; Application: XMLTransformer-1.12.13; ReportFilename: PFC_FC_LC_PFOA_WG39768-101_Form1A_FC2G_157S22_SJ1453684.html; Workgroup: WG39768; Design ID: 1722 ] Page 11 of 49 Page 1 of 1 (WG39768 -PFC_FC_LC_PFOA_WG39768-101_Form1A_FC2G_157S22_SJ1453684.html) www.axysanalytical.com AXYS METHOD MLA-043 Rev 8 Form 2 PERFLUORINATED ORGANICS ANALYSIS REPORT AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811 Contract No.: 4673 Project No. Lab Sample I.D.: CLIENT SAMPLE NO. Lab Blank Sample Collection: N/A N/A W G 39768-101 Matrix: TISSUE Sample Size: 2.00 g Sample Receipt Date: N/A Initial Calibration Date: 24-Apr-2012 Extraction Date: 23-Apr-2012 Instrument ID: LC MS/MS Analysis Date: 24-Apr-2012 Time: 23:59:06 Column ID: C18MS Extract Volume (uL): 4000 Sample Data Filename: FC2G_157 S: 22 Injection Volume (uL): 15 Blank Data Filename: F C 2 G _ 1 57 S: 22 Dilution Factor: N/A Cal. Ver. Data Filename: F C 2 G 157 S: 15 Concentration Units: ng absolute This page is part of a total report that contains information necessary for accreditation compliance. Results are compliant with NELAP accreditation described in the total report. Sample results relate only to the sample tested. LABELED COMPOUND LAB FLAG1 SPIKE CONC. CONC. FOUND R(%) 2 RETENTION TIME 13C4-PFOS 36.0 33.2 92.3 8:17 CLEANUP STANDARD 13C8-PFOA 12.0 11.2 93.7 7:04 (1) W here applicable, custom lab flags have been used on this report. (2) R(%) = percent recovery. T hese data are validated and reported a s accurate and in accord with A X Y S Analytical S e rv ice s Ltd. ISO 17025 com pliant quality assu ran ce processes. S igned :____________ B r y a n A l o n z o _________ For Axys Internal Use Only [ XSL Template: FC-Form2.xsl; Created: 27-Apr-2012 14:59:50; Application: XMLTransformer-1.12.13; ReportFilename: PFC_FC_LC_PFOA_WG39768-101_Form2_FC2G_157S22_SJ1453684.html; Workgroup: WG39768; Design ID: 1722 ] Page 12 of 49 Page 1 of 1 (WG39768 -PFC_FC_LC_PFOA_WG39768-101_Form2_FC2G_157S22_SJ1453684.html) www.axysanalytical.com AXYS METHOD MLA-043 Rev 8 Form 8A PERFLUORINATED ORGANICS ONGOING PRECISION AND RECOVERY (OPR) AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811 Contract No.: 4673 Lab Sample I.D.: W G 39768-102 Matrix: TISSUE Initial Calibration Date: 24-Apr-2012 Extraction Date: 23-Apr-2012 Instrument ID: LC MS/MS Analysis Date: 24-Apr-2012 Time: 22:51:55 Column ID: C18MS Extract Volume (uL): 4000 OPR Data Filename: FC2G_157 S: 18 Injection Volume (uL): 15 Blank Data Filename: F C 2 G _ 1 57 S: 22 Dilution Factor: N/A Cal. Ver. Data Filename: F C 2 G 157 S: 15 ALL CONCENTRATIONS REPORTED ON THIS FORM ARE CONCENTRATIONS IN EXTRACT, BASED O N A 1 m L EXTRACT VOLUME. COMPOUND LAB FLAG 1 SPIKE CONC. (ng/mL) CONC. FOUND (ng/mL) % RECOVERY RETENTION TIME PFOS 80.0 87.1 109 8:17 (1) W here applicable, custom lab flags have been used on this report. T hese data are validated and reported a s accurate and in accord with A X Y S Analytical S e rv ice s Ltd. ISO 17025 com pliant quality assu ran ce processes. S igned :____________ B r y a n A l o n z o _________ T hese pages are part of a larger report that may contain information necessary for full data evaluation. R e su lts reported relate only to the sam ple tested. For Axys Internal Use Only [ XSL Template: FC-Form8A.xsl; Created: 27-Apr-2012 14:59:50; Application: XMLTransformer-1.12.13; ReportFilename: PFC_FC_LC_PFOA_WG39768-102_Form8A_SJ1453676.html; Workgroup: WG39768; Design ID: 1722 ] Page 1 of 1 (WG39768 - PFC_FC_LC_PFOA_WG39768-102_Form8A_SJ1453676.html) Page 13 of 49 www.axysanalytical.com AXYS METHOD MLA-043 Rev 8 Form 8B PERFLUORINATED ORGANICS ONGOING PRECISION AND RECOVERY (OPR) AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811 Contract No.: 4673 Lab Sample I.D.: W G 39768-102 Matrix: TISSUE Initial Calibration Date: 24-Apr-2012 Extraction Date: 23-Apr-2012 Instrument ID: LC MS/MS Analysis Date: 24-Apr-2012 Time: 22:51:55 Column ID: C18MS Extract Volume (uL): 4000 OPR Data Filename: FC2G_157 S: 18 Injection Volume (uL): 15 Blank Data Filename: F C 2 G _ 1 57 S: 22 Dilution Factor: N/A Cal. Ver. Data Filename: F C 2 G 157 S: 15 ALL CONCENTRATIONS REPORTED ON THIS FORM ARE CONCENTRATIONS IN EXTRACT, BASED O N A 1 m L EXTRACT VOLUME. LABELED COMPOUND LAB FLAG 1 SPIKE CONC. (ng/mL) CONC. FOUND (ng/mL) % RECOVERY RETENTION TIME 13C4-PFOS 36.0 29.0 80.5 8:17 CLEANUP STANDARD 13C8-PFOA 12.0 11.2 93.3 7:04 (1) W here applicable, custom lab flags have been used on this report. T hese data are validated and reported a s accurate and in accord with A X Y S Analytical S e rv ice s Ltd. ISO 17025 com pliant quality assu ran ce processes. S igned :____________ B r y a n A l o n z o _________ T hese pages are part of a larger report that may contain information necessary for full data evaluation. R e su lts reported relate only to the sam ple tested. For Axys Internal Use Only [ XSL Template: FC-Form8B.xsl; Created: 27-Apr-2012 14:59:50; Application: XMLTransformer-1.12.13; ReportFilename: PFC_FC_LC_PFOA_WG39768-102_Form8B_SJ1453676.html; Workgroup: WG39768; Design ID: 1722 ] Page 1 of 1 (WG39768 - PFC_FC_LC_PFOA_WG39768-102_Form8B_SJ1453676.html) Page 14 of 49 www.axysanalytical.com AXYS METHOD MLA-043 Rev 8 Form 1A PERFLUORINATED ORGANICS ANALYSIS REPORT AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811 Contract No.: 4673 Project No. Lab Sample I.D.: CLIENT SAMPLE NO. MSF11D 120403 (Duplicate) Sample Collection: 03-Apr-2012 3M - ALABAM A W G39768-103 N (DUP L17838-3) Matrix: TISSUE Sample Size: 2.07 g (wet) Sample Receipt Date: 19 - A p r - 2 0 1 2 Initial Calibration Date: 24-Apr-2012 Extraction Date: 23-Apr-2012 Instrument ID: LC MS/MS Analysis Date: 25-Apr-2012 Time: 16:28:24 Column ID: C18MS Extract Volume (uL): 4000 Sample Data Filename: FC2G_158 S: 9 Injection Volume (uL): 15 Blank Data Filename: F C 2 G _ 1 57 S: 22 Dilution Factor: 5 Cal. Ver. Data Filename: F C 2 G _ 1 58 S: 4 Concentration Units: ng/g (wet weight basis) This page is part of a total report that contains information necessary for accreditation compliance. Results are compliant with NELAP accreditation described in the total report. Sample results relate only to the sample tested. COMPOUND LAB FLAG 1 CONC. FOUND REPORTING LIMIT (RL)2 RETENTION TIME PFOS D 3510 4.83 (L) 8:21 (1) W here applicable, custom lab flags have been used on this report; D = dilution data. (2) Reporting Limit (Code): S = sam ple detection limit; M = method detection limit; L = lowest calibration level equivalent; Q = contract defined limit. T hese data are validated and reported a s accurate and in accord with A X Y S Analytical S e rv ice s Ltd. ISO 17025 com pliant quality assu ran ce processes. S igned :____________ B r y a n A l o n z o _________ For Axys Internal Use Only [ XSL Template: FC-Form1A.xsl; Created: 27-Apr-2012 14:59:50; Application: XMLTransformer-1.12.13; ReportFilename: PFC_FC_LC_PFOA_WG39768-103_Form1A_FC2G_158S9_SJ1453778.html; Workgroup: WG39768; Design ID: 1722 ] Page 15 of 49 Page 1 of 1 (WG39768 -PFC_FC_LC_PFOA_WG39768-103_Form1A_FC2G_158S9_SJ1453778.html) www.axysanalytical.com AXYS METHOD MLA-043 Rev 8 Form 2 PERFLUORINATED ORGANICS ANALYSIS REPORT AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811 Contract No.: 4673 Project No. Lab Sample I.D.: CLIENT SAMPLE NO. MSF11D 120403 (Duplicate) Sample Collection: 03-Apr-2012 3M - ALABAM A W G39768-103 N (DUP L17838-3) Matrix: TISSUE Sample Size: 2.07 g (wet) Sample Receipt Date: 19-Apr-2012 Initial Calibration Date: 24-Apr-2012 Extraction Date: 23-Apr-2012 Instrument ID: LC MS/MS Analysis Date: 25-Apr-2012 Time: 16:28:24 Column ID: C18MS Extract Volume (uL): 4000 Sample Data Filename: FC2G_158 S: 9 Injection Volume (uL): 15 Blank Data Filename: F C 2 G _ 1 57 S: 22 Dilution Factor: 5 Cal. Ver. Data Filename: F C 2 G _ 1 58 S: 4 Concentration Units: ng absolute This page is part of a total report that contains information necessary for accreditation compliance. Results are compliant with NELAP accreditation described in the total report. Sample results relate only to the sample tested. LABELED COMPOUND LAB FLAG1 SPIKE CONC. CONC. FOUND R(%) 2 RETENTION TIME 13C4-PFOS D 36.0 25.9 72.0 8:21 CLEANUP STANDARD 13C8-PFOA D 12.0 10.7 89.2 7:06 (1) W here applicable, custom lab flags have been used on this report; D = dilution data. (2) R(%) = percent recovery. T hese data are validated and reported a s accurate and in accord with A X Y S Analytical S e rv ice s Ltd. ISO 17025 com pliant quality assu ran ce processes. S igned :____________ B r y a n A l o n z o _________ For Axys Internal Use Only [ XSL Template: FC-Form2.xsl; Created: 27-Apr-2012 14:59:50; Application: XMLTransformer-1.12.13; ReportFilename: PFC_FC_LC_PFOA_WG39768-103_Form2_FC2G_158S9_SJ1453778.html; Workgroup: WG39768; Design ID: 1722 ] Page 16 of 49 Page 1 of 1 (WG39768 -PFC_FC_LC_PFOA_WG39768-103_Form2_FC2G_158S9_SJ1453778.html) www.axysanalytical.com AXYS METHOD MLA-043 Rev 8 Form 1A PERFLUORINATED ORGANICS ANALYSIS REPORT AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811 Contract No.: 4673 Project No. Lab Sample I.D.: CLIENT SAMPLE NO. MSF05D 120403 (MS) Sample Collection: 03-Apr-2012 3M - ALABAM A W G39768-104 (MS) Matrix: TISSUE Sample Size: 2.01 g (wet) Sample Receipt Date: 19 - A p r - 2 0 1 2 Initial Calibration Date: 24-Apr-2012 Extraction Date: 23-Apr-2012 Instrument ID: LC MS/MS Analysis Date: 25-Apr-2012 Time: 01:56:19 Column ID: C18MS Extract Volume (uL): 4000 Sample Data Filename: FC2G_157 S: 29 Injection Volume (uL): 15 Blank Data Filename: F C 2 G _ 1 57 S: 22 Dilution Factor: N/A Cal. Ver. Data Filename: F C 2 G _ 1 57 S: 15 Concentration Units: ng/g (wet weight basis) This page is part of a total report that contains information necessary for accreditation compliance. Results are compliant with NELAP accreditation described in the total report. Sample results relate only to the sample tested. COMPOUND LAB FLAG 1 CONC. FOUND REPORTING LIMIT (RL)2 RETENTION TIME PFOS 670 0.995 (L) 8:17 (1) W here applicable, custom lab flags have been used on this report. (2) Reporting Limit (Code): S = sam ple detection limit; M = method detection limit; L = lowest calibration level equivalent; Q = contract defined limit. T hese data are validated and reported a s accurate and in accord with A X Y S Analytical S e rv ice s Ltd. ISO 17025 com pliant quality assu ran ce processes. S igned :____________ B r y a n A l o n z o _________ For Axys Internal Use Only [ XSL Template: FC-Form1A.xsl; Created: 27-Apr-2012 14:59:50; Application: XMLTransformer-1.12.13; ReportFilename: PFC_FC_LC_PFOA_WG39768-104_Form1A_FC2G_157S29_SJ1453693.html; Workgroup: WG39768; Design ID: 1722 ] Page 17 of 49 Page 1 of 1 (WG39768 -PFC_FC_LC_PFOA_WG39768-104_Form1A_FC2G_157S29_SJ1453693.html) www.axysanalytical.com AXYS METHOD MLA-043 Rev 8 Form 2 PERFLUORINATED ORGANICS ANALYSIS REPORT AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811 Contract No.: 4673 Project No. Lab Sample I.D.: CLIENT SAMPLE NO. MSF05D 120403 (MS) Sample Collection: 03-Apr-2012 3M - ALABAM A W G39768-104 (MS) Matrix: TISSUE Sample Size: 2.01 g (wet) Sample Receipt Date: 19-Apr-2012 Initial Calibration Date: 24-Apr-2012 Extraction Date: 23-Apr-2012 Instrument ID: LC MS/MS Analysis Date: 25-Apr-2012 Time: 01:56:19 Column ID: C18MS Extract Volume (uL): 4000 Sample Data Filename: FC2G_157 S: 29 Injection Volume (uL): 15 Blank Data Filename: F C 2 G _ 1 57 S: 22 Dilution Factor: N/A Cal. Ver. Data Filename: F C 2 G _ 1 57 S: 15 Concentration Units: ng absolute This page is part of a total report that contains information necessary for accreditation compliance. Results are compliant with NELAP accreditation described in the total report. Sample results relate only to the sample tested. LABELED COMPOUND LAB FLAG1 SPIKE CONC. CONC. FOUND R(%) 2 RETENTION TIME 13C4-PFOS 36.0 35.4 98.2 8:17 CLEANUP STANDARD 13C8-PFOA 12.0 11.0 91.3 7:06 (1) W here applicable, custom lab flags have been used on this report. (2) R(%) = percent recovery. T hese data are validated and reported a s accurate and in accord with A X Y S Analytical S e rv ice s Ltd. ISO 17025 com pliant quality assu ran ce processes. S igned :____________ B r y a n A l o n z o _________ For Axys Internal Use Only [ XSL Template: FC-Form2.xsl; Created: 27-Apr-2012 14:59:50; Application: XMLTransformer-1.12.13; ReportFilename: PFC_FC_LC_PFOA_WG39768-104_Form2_FC2G_157S29_SJ1453693.html; Workgroup: WG39768; Design ID: 1722 ] Page 18 of 49 Page 1 of 1 (WG39768 -PFC_FC_LC_PFOA_WG39768-104_Form2_FC2G_157S29_SJ1453693.html) www.axysanalytical.com AXYS METHOD MLA-043 Rev 8 Form 1A PERFLUORINATED ORGANICS ANALYSIS REPORT AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811 Contract No.: 4673 Project No. Lab Sample I.D.: CLIENT SAMPLE NO. MSF05D 120403 (MSD) Sample Collection: 03-Apr-2012 3M - ALABAM A WG39768-105 (MSD) Matrix: TISSUE Sample Size: 2.11 g (wet) Sample Receipt Date: 19 - A p r - 2 0 1 2 Initial Calibration Date: 24-Apr-2012 Extraction Date: 23-Apr-2012 Instrument ID: LC MS/MS Analysis Date: 25-Apr-2012 Time: 02:13:05 Column ID: C18MS Extract Volume (uL): 4000 Sample Data Filename: FC2G_157 S: 30 Injection Volume (uL): 15 Blank Data Filename: F C 2 G _ 1 57 S: 22 Dilution Factor: N/A Cal. Ver. Data Filename: F C 2 G _ 1 57 S: 15 Concentration Units: ng/g (wet weight basis) This page is part of a total report that contains information necessary for accreditation compliance. Results are compliant with NELAP accreditation described in the total report. Sample results relate only to the sample tested. COMPOUND LAB FLAG 1 CONC. FOUND REPORTING LIMIT (RL)2 RETENTION TIME PFOS 653 0.948 (L) 8:17 (1) W here applicable, custom lab flags have been used on this report. (2) Reporting Limit (Code): S = sam ple detection limit; M = method detection limit; L = lowest calibration level equivalent; Q = contract defined limit. T hese data are validated and reported a s accurate and in accord with A X Y S Analytical S e rv ice s Ltd. ISO 17025 com pliant quality assu ran ce processes. S igned :____________ B r y a n A l o n z o _________ For Axys Internal Use Only [ XSL Template: FC-Form1A.xsl; Created: 27-Apr-2012 14:59:50; Application: XMLTransformer-1.12.13; ReportFilename: PFC_FC_LC_PFOA_WG39768-105_Form1A_FC2G_157S30_SJ1453694.html; Workgroup: WG39768; Design ID: 1722 ] Page 19 of 49 Page 1 of 1 (WG39768 - PFC_FC_LC_PFOA_WG39768-105_Form1A_FC2G_157S30_SJ1453694.html) www.axysanalytical.com AXYS METHOD MLA-043 Rev 8 Form 2 PERFLUORINATED ORGANICS ANALYSIS REPORT AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811 Contract No.: 4673 Project No. Lab Sample I.D.: CLIENT SAMPLE NO. MSF05D 120403 (MSD) Sample Collection: 03-Apr-2012 3M - ALABAM A WG39768-105 (MSD) Matrix: TISSUE Sample Size: 2.11 g (wet) Sample Receipt Date: 19-Apr-2012 Initial Calibration Date: 24-Apr-2012 Extraction Date: 23-Apr-2012 Instrument ID: LC MS/MS Analysis Date: 25-Apr-2012 Time: 02:13:05 Column ID: C18MS Extract Volume (uL): 4000 Sample Data Filename: FC2G_157 S: 30 Injection Volume (uL): 15 Blank Data Filename: F C 2 G _ 1 57 S: 22 Dilution Factor: N/A Cal. Ver. Data Filename: F C 2 G _ 1 57 S: 15 Concentration Units: ng absolute This page is part of a total report that contains information necessary for accreditation compliance. Results are compliant with NELAP accreditation described in the total report. Sample results relate only to the sample tested. LABELED COMPOUND LAB FLAG1 SPIKE CONC. CONC. FOUND R(%) 2 RETENTION TIME 13C4-PFOS 36.0 37.1 103 8:17 CLEANUP STANDARD 13C8-PFOA 12.0 10.9 91.1 7:04 (1) W here applicable, custom lab flags have been used on this report. (2) R(%) = percent recovery. T hese data are validated and reported a s accurate and in accord with A X Y S Analytical S e rv ice s Ltd. ISO 17025 com pliant quality assu ran ce processes. S igned :____________ B r y a n A l o n z o _________ For Axys Internal Use Only [ XSL Template: FC-Form2.xsl; Created: 27-Apr-2012 14:59:50; Application: XMLTransformer-1.12.13; ReportFilename: PFC_FC_LC_PFOA_WG39768-105_Form2_FC2G_157S30_SJ1453694.html; Workgroup: WG39768; Design ID: 1722 ] Page 20 of 49 Page 1 of 1 (WG39768 -PFC_FC_LC_PFOA_WG39768-105_Form2_FC2G_157S30_SJ1453694.html) www.axysanalytical.com AXYS METHOD MLA-043 Rev 8 Form 3A PERFLUORINATED ORGANICS INITIAL CALIBRATION PERCENT RECOVERIES AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811 Initial Calibration Date: 24-Apr-2012 CS0 Data Filename: CS1 Data Filename: F C 2 G _ 1 57 S: 4 F C 2 G _ 1 57 S: 5 Instrument ID: LC MS/MS CS2 Data Filename: F C 2 G _ 1 57 S: 6 LC Column ID: C18MS CS3 Data Filename: F C 2 G _ 1 57 S: 7 CS4 Data Filename: F C 2 G _ 1 57 S: 8 CS5 Data Filename: F C 2 G _ 1 57 S: 9 CS6 Data Filename: F C 2 G _ 1 57 S: 10 CS7 Data Filename: F C 2 G _ 1 57 S: 11 CS8 Data Filename: N/A COMPOUND PFOS LAB FLAG1 CS0 104 CS1 91.9 CS2 86.4 PERCENT RECOVERY (%) CS3 CS4 CS5 105 99.5 112 CS6 101 CS7 99.7 CS8 (1) W here applicable, custom lab flags have been used on this report. T hese data are validated and reported a s accurate and in accord with A X Y S Analytical S e rv ice s Ltd. ISO 17025 com pliant quality assu ran ce processes. S igned :____________ K r i s t i n a C o l e m a n _________ For Axys Internal Use Only [ XSL Template: FC-Form3A.xsl; Created: 27-Apr-2012 14:59:50; Application: XMLTransformer-1.12.13; ReportFilename: PFOA_FC_LC_24-Apr-2012_FC2G__Form3A_GS45507.html; Workgroup: WG39768; Design ID: 1722 ] Page 1 of 1 (WG39768 -PFOA_FC_LC_24-Apr-2012_FC2G__Form3A_GS45507.html) Page 21 of 49 www.axysanalytical.com AXYS METHOD MLA-043 Rev 8 Form 3B PERFLUORINATED ORGANICS INITIAL CALIBRATION PERCENT RECOVERIES AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811 Initial Calibration Date: 24-Apr-2012 CS0 Data Filename: CS1 Data Filename: F C 2 G _ 1 57 S: 4 F C 2 G _ 1 57 S: 5 Instrument ID: LC MS/MS CS2 Data Filename: F C 2 G _ 1 57 S: 6 LC Column ID: C18MS CS3 Data Filename: F C 2 G _ 1 57 S: 7 CS4 Data Filename: F C 2 G _ 1 57 S: 8 CS5 Data Filename: F C 2 G _ 1 57 S: 9 CS6 Data Filename: F C 2 G _ 1 57 S: 10 CS7 Data Filename: F C 2 G _ 1 57 S: 11 CS8 Data Filename: N/A LABELED COMPOUND 13C4-PFOS CLEAN-UP STANDARD 13C8-PFOA LAB FLAG1 CS0 103 101 CS1 82.4 99.6 PERCENT RECOVERIES (%) CS2 CS3 CS4 CS5 CS6 92.7 86.9 104 103 111 94.3 98.3 101 98.9 103 CS7 117 104 CS8 (1) W here applicable, custom lab flags have been used on this report. T hese data are validated and reported a s accurate and in accord with A X Y S Analytical S e rv ice s Ltd. ISO 17025 com pliant quality assu ran ce processes. S igned :____________ K r i s t i n a C o l e m a n _________ For Axys Internal Use Only [ XSL Template: FC-Form3B.xsl; Created: 27-Apr-2012 14:59:50; Application: XMLTransformer-1.12.13; ReportFilename: PFOA_FC_LC_24-Apr-2012_FC2G__Form3B_GS45507.html; Workgroup: WG39768; Design ID: 1722 ] Page 1 of 1 (WG39768 -PFOA_FC_LC_24-Apr-2012_FC2G__Form3B_GS45507.html) Page 22 of 49 www.axysanalytical.com AXYS METHOD MLA-043 Rev 8 Form 3C PERFLUORINATED ORGANICS INITIAL CALIBRATION RETENTION TIMES AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811 Initial Calibration Date: 24-Apr-2012 CS0 Data Filename: CS1 Data Filename: F C 2 G _ 1 57 S: 4 F C 2 G _ 1 57 S: 5 Instrument ID: LC MS/MS CS2 Data Filename: F C 2 G _ 1 57 S: 6 LC Column ID: C18MS CS3 Data Filename: F C 2 G _ 1 57 S: 7 CS4 Data Filename: F C 2 G _ 1 57 S: 8 CS5 Data Filename: F C 2 G _ 1 57 S: 9 CS6 Data Filename: F C 2 G _ 1 57 S: 10 CS7 Data Filename: F C 2 G _ 1 57 S: 11 CS8 Data Filename: N/A COMPOUND PFOS LAB FLAG1 CS0 8:17 CS1 8:17 CS2 8:17 RETENTION TIMES CS3 CS4 CS5 8:17 8:17 8:17 CS6 8:17 CS7 8:17 CS8 MEAN RT 8:17 (1) W here applicable, custom lab flags have been used on this report. T hese data are validated and reported a s accurate and in accord with A X Y S Analytical S e rv ice s Ltd. ISO 17025 com pliant quality assu ran ce processes. S igned :____________ K r i s t i n a C o l e m a n _________ For Axys Internal Use Only [ XSL Template: FC-Form3C.xsl; Created: 27-Apr-2012 14:59:50; Application: XMLTransformer-1.12.13; ReportFilename: PFOA_FC_LC_24-Apr-2012_FC2G__Form3C_GS45507.html; Workgroup: WG39768; Design ID: 1722 ] Page 1 of 1 (WG39768 -PFOA_FC_LC_24-Apr-2012_FC2G__Form3C_GS45507.html) Page 23 of 49 www.axysanalytical.com AXYS METHOD MLA-043 Rev 8 Form 3D PERFLUORINATED ORGANICS INITIAL CALIBRATION RETENTION TIMES AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811 Initial Calibration Date: 24-Apr-2012 CS0 Data Filename: CS1 Data Filename: F C 2 G _ 1 57 S: 4 F C 2 G _ 1 57 S: 5 Instrument ID: LC MS/MS CS2 Data Filename: F C 2 G _ 1 57 S: 6 LC Column ID: C18MS CS3 Data Filename: F C 2 G _ 1 57 S: 7 CS4 Data Filename: F C 2 G _ 1 57 S: 8 CS5 Data Filename: F C 2 G _ 1 57 S: 9 CS6 Data Filename: F C 2 G _ 1 57 S: 10 CS7 Data Filename: F C 2 G _ 1 57 S: 11 CS8 Data Filename: N/A LABELED COMPOUND 13C4-PFOS CLEAN-UP STANDARD 13C8-PFOA LAB FLAG1 RETENTION TIMES CS0 CS1 CS2 CS3 CS4 CS5 CS6 CS7 8:17 8:17 8:17 8:17 8:21 8:17 8:17 8:17 CS8 MEAN RT 8:18 7:04 7:04 7:04 7:04 7:04 7:04 7:04 7:04 7:04 (1) W here applicable, custom lab flags have been used on this report. T hese data are validated and reported a s accurate and in accord with A X Y S Analytical S e rv ice s Ltd. ISO 17025 com pliant quality assu ran ce processes. S igned :____________ K r i s t i n a C o l e m a n _________ For Axys Internal Use Only [ XSL Template: FC-Form3D.xsl; Created: 27-Apr-2012 14:59:50; Application: XMLTransformer-1.12.13; ReportFilename: PFOA_FC_LC_24-Apr-2012_FC2G__Form3D_GS45507.html; Workgroup: WG39768; Design ID: 1722 ] Page 1 of 1 (WG39768 -PFOA_FC_LC_24-Apr-2012_FC2G__Form3D_GS45507.html) Page 24 of 49 www.axysanalytical.com AXYS METHOD MLA-043 Rev 8 Form 4A PERFLUORINATED ORGANICS CALIBRATION VERIFICATION AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811 Initial Calibration Date: 24-Apr-2012 VER Data Filename: F C 2 G _ 1 57 S: 15 Instrument ID: LC MS/MS Analysis Date: 24-Apr-2012 LC Column ID: C18MS Analysis Time: 2 2 : 0 1 :37 COMPOUND PFOS LAB FLAG1 RETENTION TIME 8:17 EXPECTED CONC. (ng) 80.0 CONC. FOUND (ng) 86.8 RECOVERY (%) 108 (1) W here applicable, custom lab flags have been used on this report. T hese data are validated and reported a s accurate and in accord with A X Y S Analytical S e rv ice s Ltd. ISO 17025 com pliant quality assu ran ce processes. S igned :____________ B r y a n A l o n z o _________ For Axys Internal Use Only [ XSL Template: FC-Form4A.xsl; Created: 27-Apr-2012 14:59:50; Application: XMLTransformer-1.12.13; ReportFilename: PFOA _FC_LC_FC2GJ57S15_Form4A_SJ1453671.html; Workgroup: WG39768; Design ID: 1722 ] Page 1 of 1 (WG39768 -PFOA_FC_LC_FC2G_157S15__Form4A_SJ1453671.html) Page 25 of 49 www.axysanalytical.com AXYS METHOD MLA-043 Rev 8 Form 4B PERFLUORINATED ORGANICS CALIBRATION VERIFICATION AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811 Initial Calibration Date: 24-Apr-2012 VER Data Filename: F C 2 G _ 1 57 S: 15 Instrument ID: LC MS/MS Analysis Date: 24-Apr-2012 LC Column ID: C18MS Analysis Time: 2 2 : 0 1 :37 LABELED COMPOUND 13C4-PFOS CLEANUP STANDARD 13C8-PFOA LAB FLAG 1 RETENTION TIME 8:17 EXPECTED CONC. (ng) 36.0 CONC.FOUND (ng) 36.0 R E C O V E R Y (%) 100 7:06 12.0 12.1 101 (1) W here applicable, custom lab flags have been used on this report. T hese data are validated and reported a s accurate and in accord with A X Y S Analytical S e rv ice s Ltd. ISO 17025 com pliant quality assu ran ce processes. S igned :____________ B r y a n A l o n z o _________ For Axys Internal Use Only [ XSL Template: FC-Form4B.xsl; Created: 27-Apr-2012 14:59:50; Application: XMLTransformer-1.12.13; ReportFilename: PFOA _FC_LC_FC2GJ57S15_Form4B_SJ1453671.html; Workgroup: WG39768; DesignID: 1722 ] Page 1 of 1 (WG39768 -PFOA_FC_LC_FC2G_157S15__Form4B_SJ1453671.html) Page 26 of 49 www.axysanalytical.com AXYS METHOD MLA-043 Rev 8 Form 4A PERFLUORINATED ORGANICS CALIBRATION VERIFICATION AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811 Initial Calibration Date: 24-Apr-2012 VER Data Filename: F C 2 G _ 1 58 S: 4 Instrument ID: LC MS/MS Analysis Date: 25-Apr-2012 LC Column ID: C18MS Analysis Time: 15:04:34 COMPOUND PFOS LAB FLAG1 RETENTION TIME 8:17 EXPECTED CONC. (ng) 80.0 CONC. FOUND (ng) 81.1 RECOVERY (%) 101 (1) W here applicable, custom lab flags have been used on this report. T hese data are validated and reported a s accurate and in accord with A X Y S Analytical S e rv ice s Ltd. ISO 17025 com pliant quality assu ran ce processes. S igned :____________ B r y a n A l o n z o _________ For Axys Internal Use Only [ XSL Template: FC-Form4A.xsl; Created: 27-Apr-2012 14:59:50; Application: XMLTransformer-1.12.13; Report Filename: PFOA_FC_LC_FC2G_158S4__Form4A_SJ1453773.html; Workgroup: WG39768; Design ID: 1722 ] Page 1 of 1 (WG39768 -PFOA_FC_LC_FC2G_158S4__Form4A_SJ1453773.html) Page 27 of 49 www.axysanalytical.com AXYS METHOD MLA-043 Rev 8 Form 4B PERFLUORINATED ORGANICS CALIBRATION VERIFICATION AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811 Initial Calibration Date: 24-Apr-2012 VER Data Filename: F C 2 G _ 1 58 S: 4 Instrument ID: LC MS/MS Analysis Date: 25-Apr-2012 LC Column ID: C18MS Analysis Time: 15:04:34 LABELED COMPOUND 13C4-PFOS CLEANUP STANDARD 13C8-PFOA LAB FLAG 1 RETENTION TIME 8:21 EXPECTED CONC. (ng) 36.0 CONC.FOUND (ng) 28.3 RECOVERY (%) 78.6 7:06 12.0 11.8 98.3 (1) W here applicable, custom lab flags have been used on this report. T hese data are validated and reported a s accurate and in accord with A X Y S Analytical S e rv ice s Ltd. ISO 17025 com pliant quality assu ran ce processes. S igned :____________ B r y a n A l o n z o _________ For Axys Internal Use Only [ XSL Template: FC-Form4B.xsl; Created: 27-Apr-2012 14:59:50; Application: XMLTransformer-1.12.13; ReportFilename: PFOA_FC_LC_FC2G_158S4__Form4B_SJ1453773.html; Workgroup: WG39768; Design ID: 1722 ] Page 1 of 1 (WG39768 -PFOA_FC_LC_FC2G_158S4__Form4B_SJ1453773.html) Page 28 of 49 www.axysanalytical.com AXYS METHOD MLA-043 Rev 8 PERFLUORINATED ORGANICS ANALYSIS REPORT RELATIVE PERCENT DIFFERENCE AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811 Project No. Contract No.: 4673 Client ID: MSF11D 120403 Concentration Units: 3M - ALABAM A ng/g (wet weight basis) COMPOUND PFOS L17838-3 (A)_______ LAB FLAG1 CONC. FOUND D 3720 WG39768-103 LAB FLAG 1 CONC. FOUND D 3510 MEAN 3620 RELATIVE PERCENT DIFFERENCE 5.77 (1) W here applicable, custom lab flags have been used on this report; D = dilution data. T hese data are validated and reported a s accurate and in accord with A X Y S Analytical S e rv ice s Ltd. ISO 17025 com pliant quality assu ran ce processes. S igned :____________ B r y a n A l o n z o _________ T hese pages are part of a larger report that may contain information necessary for full data evaluation. Results reported relate only to the sam ple tested. For Axys Internal Use Only [ XSL Template: RPD.xsl; Created: 27-Apr-2012 15:00:24; Application: XMLTransformer-1.12.13; ReportFilename: RPD_FC_LC_PFOA-RPD_WG39768-103_L17838-3_.html; Workgroup: WG39768; Design ID: 1722 ] Page 1 of 1 (WG39768 - RPD_FC_LC_PFOA-RPD_WG39768-103_L17838-3_.html) Page 29 of 49 www.axysanalytical.com AXYS METHOD MLA-043 Rev 8 AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811 Contract No.: 4673 Form 8C PERFLUORINATED ORGANICS MATRIX SPIKE (MS) ANALYSIS REPORT Project No. Lab Sample I.D.: Matrix: TISSUE Sample Size: Extraction Date: 23-Apr-2012 Initial Calibration Date: Analysis Date: 25-Apr-2012 Time: 01:56:19 Instrument ID: Extract Volume (uL): 4000 Column ID: Injection Volume (uL): 15 MS Data Filename: Dilution Factor: N/A Blank Data Filename: Cal. Ver. Data Filename: CLIENT SAMPLE NO. MSF05D 120403 (MS) Sample Collection: 03-Apr-2012 3M - ALABAM A W G39768-104 (MS) 2.01 g (wet) 24-Apr-2012 LC MS/MS C18MS FC2G_157 S: 29 F C 2 G _ 1 57 S: 22 F C 2 G 157 S: 15 ALL CONCENTRATIONS REPORTED ON THIS FORM ARE CONCENTRATIONS ON SAMPLE SIZE BASIS COMPOUND PFOS LAB FLAG1 ION ABUND. RATIO N/A SPIKE CONC (ng/g) 39.8 SAMPLE LAB FLAG 1 SAMPLE CONC (ng/g) 656 CONC. FOUND (ng/g) 670 MS R% 2 36.4 REL % DIFF (1) W here applicable, custom lab flags have been used on this report. (2) R% = percent recovery. T hese data are validated and reported a s accurate and in accord with A X Y S Analytical S e rv ice s Ltd. ISO 17025 com pliant quality assu ran ce processes. S igned :____________ B r y a n A l o n z o _________ T hese pages are part of a larger report that may contain information necessary for full data evaluation. Results reported relate only to the sam ple tested. For Axys Internal Use Only [ XSL Template: MS.xsl; Created: 27-Apr-2012 15:00:16; Application: XMLTransformer-1.12.13; Report Filename: MS_FC_LC_PFOA-MS-MSD_WG39768-104_L17838-2_Form8C.html; Workgroup: WG39768; Design ID: 1722 ] Page 1 of 1 (WG39768 - MS_FC_LC_PFOA-MS-MSD_WG39768-104_L17838-2_Form8C.html) Page 30 of 49 www.axysanalytical.com AXYS METHOD MLA-043 Rev 8 Form 8E PERFLUORINATED ORGANICS MATRIX SPIKE DUPLICATE (MSD) ANALYSIS REPORT AXYS ANALYTICAL SERVICES 2045 MILLS RD., SIDNEY, B.C., CANADA V8L 5X2 TEL (250) 655-5800 FAX (250) 655-5811 Project No. Contract No.: 4673 Lab Sample I.D.: Matrix: TISSUE Sample Size: Extraction Date: 23-Apr-2012 Initial Calibration Date: Analysis Date: 25-Apr-2012 Time: 02:13:05 Instrument ID: Extract Volume (uL): 4000 Column ID: Injection Volume (uL): 15 MSD Data Filename: Dilution Factor: N/A Blank Data Filename: Cal. Ver. Data Filename: CLIENT SAMPLE NO. MSF05D 120403 (MSD) Sample Collection: 03-Apr-2012 3M - ALABAM A W G39768-105 (MSD) 2.11 g (wet) 24-Apr-2012 LC MS/MS C18MS FC2G_157 S: 30 F C 2 G _ 1 57 S: 22 F C 2 G 157 S: 15 ALL CONCENTRATIONS REPORTED ON THIS FORM ARE CONCENTRATIONS ON SAMPLE SIZE BASIS COMPOUND PFOS LAB FLAG1 ION ABUND. RATIO N/A SPIKE CONC (ng/g) 37.9 SAMPLE LAB FLAG 1 SAMPLE CONC (ng/g) 656 CONC. FOUND (ng/g) 653 MSD R% 2 REL % DIFF (1) W here applicable, custom lab flags have been used on this report. (2) R% = percent recovery. T hese data are validated and reported a s accurate and in accord with A X Y S Analytical S e rv ice s Ltd. ISO 17025 com pliant quality assu ran ce processes. S igned :____________ B r y a n A l o n z o _________ T hese pages are part of a larger report that may contain information necessary for full data evaluation. Results reported relate only to the sam ple tested. For Axys Internal Use Only [ XSL Template: MS.xsl; Created: 27-Apr-2012 15:00:16; Application: XMLTransformer-1.12.13; Report Filename: MS_FC_LC_PFOA-MS-MSD_WG39768-105_L17838-2_Form8E.html; Workgroup: WG39768; Design ID: 1722 ] Page 1 of 1 (WG39768 - MS_FC_LC_PFOA-MS-MSD_WG39768-105_L17838-2_Form8E.html) Page 31 of 49 www.axysanalytical.com AXYS Analytical Services Ltd. Table 1a NELAP Accreditation Held by AXYS Analytical Services Ltd. for Chlorinated Dioxins/Furans, Chlorinated Pesticides, PCBs and PAHs Matrix Codes for Table 1a NPW = Non-Potable Water DrW = Drinking Water S = Solid T = Tissue Accreditation Method Codes and Explanation for Table 1 Code Accreditation Certificate Applicable AXYS Method and Description No. Method Reference 1 EPA 1613B MLA-017, performance based implementation of EPA1613B (GC/HRMS) 2 EPA 8290 MLA-017, performance based implementation of EPA 8290 (Gc /HRMS) 3 AXYS MLA-017 MLA-017, performance based implementation of EPA 1613B, 8290 (GC/HRMS) 4 EPA 608 MLA-007, performance based implementation of EPA 608 (GC/ECD) 5 EPA 8270C or 8270D MLA-007, performance based modification of 8270C/D (GC/LRMS) 6 EPA8081Aor8081B MLA-007, performance based implementation of EPA 8081A/B (GC/ECD) 7 EPA 1668A MLA-010, performance based implementation of EPA 1668A (GC/HRMS) 8 SM 6630B MLA-007, performance based implementation of SM 18-20 6630B (GC/ECD) 9 EPA 1625B MLA-021, performance based modification of EPA 1625B (GC/LRMS) 11 EPA 625 MLA-007, performance based modification of EPA 625 (g C/LRMS) 20 EPA 8270C or 8270D MLA-021, performance based modification of EPA 8270C/D (GC/LRMS) TABLE 1 PCDD/F - Polychlorinated Dioxins and Furans Dioxins Dioxins and Dibenzofurans 1,2,3,4,6,7,8-HpCDD 1,2,3,4,6,7,8-HpCDF 1,2,3,4,7,8,9-HpCDF 1,2,3,4,7,8-HxCDD 1,2,3,4,7,8-HxCDF 1,2,3,6,7,8-HxCDD ACC-101 Rev 06, 29-Jun-2011 New York State Department of Health Lab ID 11674 NELAP Primary NP W S California Department of Public Health Lab ID 01138CA NELAP Secondary NP W S State of Florida Department of Health Lab ID E871007 NELAP Primary Dr. NP WW S T 1 2 12 12 12 12 12 12 1, 2, 3 1, 2, 3 1, 2, 3 1, 2, 3 1, 2, 3 1, 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 State of New Jersey Department of Environmental Protection Lab ID CANA005 NELAP Secondary Dr. NP WW S T 12 2 12 2 12 2 12 2 12 2 12 2 Page 1 of 18 Page 32 of 49 www.axysanalytical.com TABLE 1 1,2,3,6,7,8-HxCDF 1,2,3,7,8,9-HxCDD 1,2,3,7,8,9-HxCDF 1,2,3,7,8-PeCDD 1,2,3,7,8-PeCDF 2,3,4,6,7,8-HxCDF 2,3,4,7,8-PeCDF 2,3,7,8-TCDD 2,3,7,8-TCDF OCDD OCDF Total TCDD Total TCDF Total PeCDD Total PeCDF Total HxCDD Total HxCDF Total HpCDD Total HpCDF PCBs - Polychlorinated biphenyls PCB 1 2-Chlorobiphenyl PCB 3 4-Chlorobiphenyl PCB 4 2,2'-Dichlorobiphenyl PCB 5 2,3-Dichlorobiphenyl PCB 15 4,4'-Dichlorobiphenyl PCB 18 2,2',5-Trichlorobiphenyl PCB 19 2,2',6-Trichlorobiphenyl PCB 31 2,4',5-Trichlorobiphenyl PCB 37 3,4,4'-Trichlorobiphenyl PCB 44 2,2',3,5'-Tetrachlorobiphenyl PCB 52 2,2',5,5'-Tetrachlorobiphenyl PCB 54 2,2',6,6'-Tetrachlorobiphenyl PCB 66 2,3',4,4'-Tetrachlorobiphenyl PCB 77 3,3',4,4'-Tetrachlorobiphenyl PCB 81 3,4,4',5-Tetrachlorobiphenyl PCB 87 2,2',3,4,5'-Pentachlorobiphenyl ACC-101 Rev 06, 29-Jun-2011 AXYS Analytical Services Ltd. New York State Department of Health Lab ID 11674 NELAP Primary NP W S 1 California Department of Public Health Lab ID 01138CA NELAP Secondary NP W S 12 12 12 12 12 12 12 12 12 12 12 1 1 1 1 1 1 1 1 State of Florida Department of Health Lab ID E871007 NELAP Primary Dr. NP WW S T 1, 2, 3 1, 2, 3 1, 2, 3 1, 2, 3 1, 2, 3 1, 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 1, 2, 3 1, 2, 3 1, 2, 3 1, 2, 3 1, 2, 3 1, 2, 3 1, 2, 3 1, 2, 3 1, 2, 3 1, 2, 3 1, 2, 3 1, 2, 3 1, 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 2, 3 State of New Jersey Department of Environmental Protection Lab ID CANA005 NELAP Secondary Dr. NP WW 1 1 1 1 1 1 1 1 1 1 1 S 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 T 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 Page 2 of 18 Page 33 of 49 www.axysanalytical.com AXYS Analytical Services Ltd. TABLE 1 PCB 101 PCB 104 PCB 105 PCB 109 PCB 114 PCB 118 PCB 123 PCB 124 PCB 126 PCB 138 PCB 141 PCB 151 PCB 153 PCB 155 PCB 156 PCB 157 PCB 167 PCB 169 PCB 170 PCB 180 PCB 183 PCB 187 PCB 188 PCB 189 PCB 202 PCB 205 PCB 206 PCB 208 PCB 209 Aroclor 1260 Aroclor 1254 Aroclor 1221 Aroclor 1232 Aroclor 1248 Aroclor 1016 Aroclor 1242 2,2',4,5,5'-Pentachlorobiphenyl 2,2',4,6,6'-Pentachlorobiphenyl 2,3,3',4,4'-Pentachlorobiphenyl 2,3,3',4,6-Pentachlorobiphenyl 2,3,4,4',5-Pentachlorobiphenyl 2,3',4,4',5-Pentachlorobiphenyl 2,3',4,4',5'-Pentachlorobiphenyl 2,3',4',5,5'-Pentachlorobiphenyl 3,3',4,4',5-Pentachlorobiphenyl 2,2',3,4,4',5'-Hexachlorobiphenyl 2,2',3,4,5,5'-Hexachlorobiphenyl 2,2',3,5,5',6-Hexachlorobiphenyl 2,2',4,4',5,5'-Hexachlorobiphenyl 2,2',4,4',6,6'-Hexachlorobiphenyl 2,3,3',4,4',5-Hexachlorobiphenyl 2,3,3',4,4',5'-Hexachlorobiphenyl 2,3',4,4',5,5'-Hexachlorobiphenyl 3,3',4,4',5,5'-Hexachlorobiphenyl 2,2',3,3',4,4',5-Heptachlorobiphenyl 2,2',3,4,4',5,5'-Heptachlorobiphenyl 2,2',3,4,4',5',6-Heptachlorobiphenyl 2,2',3,4',5,5',6-Heptachlorobiphenyl 2,2',3,4',5,6,6'-Heptachlorobiphenyl 2,3,3',4,4',5,5'-Heptachlorobiphenyl 2,2',3,3',5,5',6,6'-Octachlorobiphenyl 2,3,3',4,4',5,5',6-Octachlorobiphenyl 2,2',3,3',4,4',5,5',6-Nonachlorobiphenyl 2,2',3,3',4,5,5',6,6'-Nonachlorobiphenyl Decachlorobiphenyl New York State Department of Health Lab ID 11674 NELAP Primary NP W S 77 77 77 77 77 77 77 California Department of Public Health Lab ID 01138CA NELAP Secondary NP W S 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7, 11 7, 11 7, 11 7, 11 7, 11 7, 11 7, 11 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 5, 7 5, 7 5, 7 5, 7 5, 7 5, 7 5, 7 11 11 11 11 11 11 11 5 5 5 5 5 5 5 State of Florida Department of Health Lab ID E871007 NELAP Primary Dr. NP WW S T ACC-101 Rev 06, 29-Jun-2011 State of New Jersey Department of Environmental Protection Lab ID CANA005 NELAP Secondary Dr. NP WW 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 S 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 T Page 3 of 18 Page 34 of 49 www.axysanalytical.com TABLE 1 Pesticides 4,4'-DDD 4,4'-DDE 4,4'-DDT Aldrin Alpha-HCH Beta-HCH cis-Chlordane (alpha-Chlordane) Chlordane, technical Delta-HCH Dieldrin Endosulphan I Endosulphan II Endosulphan sulphate Endrin Endrin aldehyde trans-Chlordane (gamma-Chlordane) Gamma-HCH (Lindane) Heptachlor Heptachlor epoxide Hexachlorobenzene Methoxychlor Mi rex PAH Anthracene Pyrene Benzo[ghilperylene Indeno[1,2,3-cdlpyrene Benzo[blfluoranthene Fluoranthene Benzo[klfluoranthene Acenaphthylene Chrysene Benzo[a]pyrene Dibenz[ahlanthracene Benz[alanthracene ACC-101 Rev 06, 29-Jun-2011 AXYS Analytical Services Ltd. New York State Department of Health Lab ID 11674 NELAP Primary NP W S California Department of Public Health Lab ID 01138CA NELAP Secondary NP W S State of Florida Department of Health Lab ID E871007 NELAP Primary Dr. NP WW S T 11 5 11 5 11 5 11 5 11 5 11 5 11 5 11 5 11 5 11 5 11 5 11 5 55 5, 11 5 11 5 11 5 11 5 4646 4646 4646 4646 4646 4646 55 11 5 11 5 11 5 11 5 4646 9595 4,8 6 8 6 5 State of New Jersey Department of Environmental Protection Lab ID CANA005 NELAP Secondary Dr. NP WW S T 9 20 9 20 9 20 9 20 9 20 9 20 9 20 9 20 9 20 9 20 9 20 9 20 9 20 9 20 9 20 9 20 9 20 9 20 9 20 9 20 9 20 9 20 9 20 9 20 Page 4 of 18 Page 35 of 49 www.axysanalytical.com TABLE 1 Acenaphthene Phenanthrene Fluorene Naphthalene AXYS Analytical Services Ltd. New York State Department of Health Lab ID 11674 NELAP P rim a ry NP W S 9 20 9 20 9 20 9 20 California Department of Public Health Lab ID 01138CA NELAP Secondary NP W S 9 20 9 20 9 20 9 20 State of Florida Department of Health Lab ID E871007 NELAP Primary Dr. NP WW S T State of New Jersey Department of Environmental Protection Lab ID C AN A005 NELAP Secondary Dr. NP WW S T ACC-101 Rev 06, 29-Jun-2011 Page 5 of 18 Page 36 of 49 www.axysanalytical.com AXYS Analytical Services Ltd. Table 1b NELAP Accreditation Held by AXYS Analytical Services Ltd. for Perfluorinated Organic Compounds Matrix Codes for Table 1b NPW = Non-Potable Water DrW = Drinking Water S = Solid T = Tissue Accreditation Method Codes and Explanation for Table 1b Code Accreditation Certificate Applicable AXYS Method and Description No. Method Reference 12 AXYS MLA-041 13 AXYS MLA-043 14 AXYS MLA-060 MLA-041, laboratory performance based method (LC/MS-MS) MLA-043, laboratory performance based method (LC/MS-MS) MLA-060, laboratory performance based method (LC/MS-MS) TABLE 1 State of Florida Department of Health Lab ID E871007 NELAP Primary Dr. NP WW S T Minnesota Department of Health Lab ID 232-999-430 NELAP Primary! Dr. NP WW S T State of New Jersey Department of Environmental Protection Lab ID CANA005 NELAP Secondary Dr. NP WW S T PFC - Perfluorinated Organic Compounds Perfluorobutanoate (PFBA) Note Perfluoropentanoate (PFPeA) Note Perfluorohexanoate (PFHxA) Note Perfluoroheptanoate (PFHpA) Note Perfluorooctanoate (PFOA) Nte Perfluorononanoate (PFNA) Note Perfluorodecanoate (PFDA) Note Perfluoroundecanoate (PFUnA) N ote N Perfluorododecanoate (PFDoA) N ote N Perfluorobutanesulfonate (PFBS) Perfluorohexanesulfonate (PFHxS) Perfluorooctanesulfonate (PFOS) Perfluorooctane sulfonamide (PFOSA) 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 14 14 12 13 Note: Accreditations by Minnesota Department of Health and New Jersey Department of Environmental Protection are against the corresponding acid form of the anion shown. ACC-101 R ev 06, 29-Jun-2011 P age 6 o f 18 Page 37 of 49 www.axysanalytical.com AXYS Analytical Services Ltd. Table 2: Canadian and US State Specific Accreditation Held by AXYS Analytical Services Ltd. Matrix Codes for Table 2 NP W = Non-Potable Water Dr. W = Drinking W ater W = Aqueous S = Solid T = Tissue Accreditation Method Codes and Explanation for Table 2 Code Accreditation Certificate No. Method Reference Applicable AXYS Method and Description 1 EPA 1613 3 AXYS MLA-017 7 EPA 1668A 10 AXYS MLA-007 12 AXYS MLA-041 13 AXYS MLA-043 14 AXYS MLA-060 15 AXYS MLA-010 16 AXYS MLA-028 17 AXYS MLA-033 18 AXYS MLA-021 19 AXYS MLA-075 MLA-017 Performance based implementation of EPA 1613B (GC/HRMS) MLA-017 Performance based implementation of EPA 1613B (g C/HRMs ) MLA-010 Performance based implementation of EPA 1668A (GC/HRMs ) MLA-007, Performance based modification of EPA 8270C/D, 8081A/B (GC/LRMS and GC/ECD) MLA-041 Laboratory performance based method (LC/MS-MS) MLA-043 Laboratory performance based method (LC/MS-MS) MLA-060 Laboratory performance based method (LC/MS-MS) MLA-010 Performance based implementation of EPA 1668A (GC/HRMS) MLA-028 Laboratory performance based method (GC/HRMS) MLA-033 Performance based implementation of EPA 1614 (Gc /HRMS) MLA-021 Performance based modification of EPA 8270C/D (GC/LRMS) MLA-075 Performance based implementation of EPA 1694 (LC/MS-MS) TABLE 2 PCDD/F - Polychlorinated Dioxins and Furans 1,2,3,4,6,7,8-HpCDD 1,2,3,4,6,7,8-HpCDF 1,2,3,4,7,8,9-HpCDF 1,2,3,4,7,8-HxCDD 1,2,3,4,7,8-HxCDF 1,2,3,6,7,8-HxCDD 1,2,3,6,7,8-HxCDF 1,2,3,7,8,9-HxCDD 1,2,3,7,8,9-HxCDF 1,2,3,7,8-PeCDD 1,2,3,7,8-PeCDF 2,3,4,6,7,8-HxCDF 2,3,4,7,8-PeCDF 2,3,7,8-TCDD 2,3,7,8-TCDF OCDD OCDF Total TCDD Total TCDF Total PeCDD Total PeCDF Total HxCDD ACC-101 Rev 06, 29-Jun-2011 Canadian Association for Laboratory Accreditation (CALA) Accreditation No.: A 2637 W S Pulp T 3333 3333 3333 3333 3333 3333 3333 3333 3333 3333 3333 3333 3333 3333 3333 3333 3333 Washington State Department of Ecology Lab. ID: C404 NP S W 11 11 11 11 11 11 11 11 11 11 11 11 11 11 11 11 11 11 11 11 11 11 Page 7 of 18 Page 38 of 49 www.axysanalytical.com AXYS Analytical Services Ltd. TABLE 2 Total HxCDF Total HpCDD Total HpCDF Total PCDD Total PCDF Total PCDD + PCDF PCBs - Polychlorinated biphenyls PCB 1 2-Chlorobiphenyl PCB 2 3-Chlorobiphenyl PCB 3 4-Chlorobiphenyl PCB 4 2,2'-Dichlorobiphenyl PCB 5 2,3-Dichlorobiphenyl PCB 6 2,3'-Dichlorobiphenyl PCB 7 2,4-Dichlorobiphenyl PCB 8 2,4'-Dichlorobiphenyl PCB 8/5 PCB 9 2,5-Dichlorobiphenyl PCB 10 2,6-Dichlorobiphenyl PCB 11 3,3'-Dichlorobiphenyl PCB 12 3,4-Dichlorobiphenyl PCB 13 3,4'-Dichlorobiphenyl PCB 14 3,5-Dichlorobiphenyl PCB 15 4,4'-Dichlorobiphenyl PCB 16 2,2',3-T richlorobiphenyl PCB 16/32 PCB 17 2,2',4-T richlorobiphenyl PCB 18 2,2',5-Trichlorobiphenyl PCB 19 2,2',6-T richlorobiphenyl PCB 20 2,3,3'-T richlorobiphenyl PCB 21 2,3,4-T richlorobiphenyl PCB 22 2,3,4'-T richlorobiphenyl PCB 23 2,3,5-T richlorobiphenyl PCB 24 2,3,6-T richlorobiphenyl PCB 24/27 PCB 25 2,3',4-T richlorobiphenyl PCB 26 2,3',5-Trichlorobiphenyl PCB 27 2,3',6-T richlorobiphenyl PCB 28 2,4,4'-T richlorobiphenyl PCB 29 2,4,5-T richlorobiphenyl PCB 30 2,4,6-T richlorobiphenyl PCB 31 2,4',5-Trichlorobiphenyl PCB 32 2,4',6-T richlorobiphenyl PCB 33 2,3',4'-T richlorobiphenyl PCB 33/20/21 PCB 34 2,3',5'-T richlorobiphenyl PCB 35 3,3',4-T richlorobiphenyl PCB 36 3,3',5-T richlorobiphenyl PCB 37 3,4,4'-T richlorobiphenyl PCB 38 3,4,5-T richlorobiphenyl PCB 39 3,4',5-T richlorobiphenyl PCB 40 2,2',3,3'-Tetrachlorobiphenyl PCB 41 2,2',3,4-Tetrachlorobiphenyl PCB 41/71/64/68 Canadian Association for Laboratory Accreditation (CALA) Accreditation No.: A 2637 W S Pulp T Washington State Department of Ecology Lab. ID: C404 NP S W 11 11 11 11 11 11 15 15 15 15 15 15 15 15 10 15 15 15 15 15 15 10, 15 15 10 10, 15 10, 15 10, 15 15 15 10, 15 15 15 10 10, 15 10, 15 15 10, 15 15 15 10, 15 15 15 18 15 15 15 15 15 15 10, 15 15 10 15 15 15 15 15 15 15 15 10 15 15 15 15 15 15 10, 15 15 10 10, 15 10, 15 10, 15 15 15 10, 15 15 15 10 10, 15 10, 15 15 10, 15 15 15 10, 15 15 15 10 15 15 15 15 15 15 10, 15 15 10 15 15 15 15 15 15 15 15 10 15 15 15 15 15 15 10, 15 15 10 10, 15 10, 15 10, 15 15 15 10, 15 15 15 10 10, 15 10, 15 15 10, 15 15 15 10, 15 15 15 10 15 15 15 15 15 15 10, 15 15 10 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 ACC-101 Rev 06, 29-Jun-2011 Page 8 of 18 Page 39 of 49 www.axysanalytical.com AXYS Analytical Services Ltd. TABLE 2 PCB 42 PCB 42/59 PCB 43 PCB 44 PCB 45 PCB 46 PCB 47 PCB 47/48/75 PCB 48 PCB 49 PCB 49/43 PCB 50 PCB 51 PCB 52 PCB 52/73 PCB 53 PCB 54 PCB 55 PCB 56 PCB 56/60 PCB 57 PCB 58 PCB 59 PCB 60 PCB 61 PCB 62 PCB 63 PCB 64 PCB 65 PCB 66 PCB 66/80 PCB 67 PCB 68 PCB 69 PCB 70 PCB 70/76 PCB 71 PCB 72 PCB 73 PCB 74 PCB 74/61 PCB 75 PCB 76 PCB 77 PCB 78 PCB 79 PCB 80 PCB 81 PCB 82 PCB 83 PCB 83/108 PCB 84 PCB 85 PCB 85/120 2,2',3,4'-Tetrachlorobiphenyl 2,2',3,5-Tetrachlorobiphenyl 2,2',3,5'-Tetrachlorobiphenyl 2,2',3,6-Tetrachlorobiphenyl 2,2',3,6'-Tetrachlorobiphenyl 2,2',4,4'-Tetrachlorobiphenyl 2,2',4,5-Tetrachlorobiphenyl 2,2',4,5'-Tetrachlorobiphenyl 2,2',4,6-Tetrachlorobiphenyl 2,2',4,6'-Tetrachlorobiphenyl 2,2',5,5'-Tetrachlorobiphenyl 2,2',5,6'-Tetrachlorobiphenyl 2,2',6,6'-Tetrachlorobiphenyl 2,3,3',4-Tetrachlorobiphenyl 2,3,3',4'-Tetrachlorobiphenyl 2,3,3',5-Tetrachlorobiphenyl 2,3,3',5'-Tetrachlorobiphenyl 2,3,3',6-Tetrachlorobiphenyl 2,3,4,4'-Tetrachlorobiphenyl 2,3,4,5-Tetrachlorobiphenyl 2,3,4,6-Tetrachlorobiphenyl 2,3,4',5-Tetrachlorobiphenyl 2,3,4',6-Tetrachlorobiphenyl 2,3,5,6-Tetrachlorobiphenyl 2,3',4,4'-Tetrachlorobiphenyl 2,3',4,5-Tetrachlorobiphenyl 2,3',4,5'-Tetrachlorobiphenyl 2,3',4,6-Tetrachlorobiphenyl 2,3',4',5-Tetrachlorobiphenyl 2,3',4',6-Tetrachlorobiphenyl 2,3',5,5'-Tetrachlorobiphenyl 2,3',5',6-Tetrachlorobiphenyl 2,4,4',5-Tetrachlorobiphenyl 2,4,4',6-Tetrachlorobiphenyl 2,3',4',5'-Tetrachlorobiphenyl 3,3',4,4'-Tetrachlorobiphenyl 3,3',4,5-Tetrachlorobiphenyl 3,3',4,5'-Tetrachlorobiphenyl 3,3',5,5'-Tetrachlorobiphenyl 3,4,4',5-Tetrachlorobiphenyl 2,2',3,3',4-Pentachlorobiphenyl 2,2',3,3',5-Pentachlorobiphenyl 2,2',3,3',6-Pentachlorobiphenyl 2,2',3,4,4'-Pentachlorobiphenyl Canadian Association for Laboratory Accreditation (CALA) Accreditation No.: A 2637 W S Pulp T 15 10 15 10, 15 10, 15 10, 15 15 10 15 15 10 15 15 15 10 15 15 15 15 10 15 15 15 15 15 15 15 15 15 15 10 15 15 15 15 10 15 15 15 15 10 15 15 10, 15 15 15 15 15 15 15 10 10, 15 15 10 15 10 15 10, 15 10, 15 10, 15 15 10 15 15 10 15 15 15 10 15 15 15 15 10 15 15 15 15 15 15 15 15 15 15 10 15 15 15 15 10 15 15 15 15 10 15 15 10, 15 15 15 15 15 15 15 10 10, 15 15 10 15 10 15 10, 15 10, 15 10, 15 15 10 15 15 10 15 15 15 10 15 15 15 15 10 15 15 15 15 15 15 15 15 15 15 10 15 15 15 15 10 15 15 15 15 10 15 15 10, 15 15 15 15 15 15 15 10 10, 15 15 10 Washington State Department of Ecology Lab. ID: C404 NP S W 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 ACC-101 Rev 06, 29-Jun-2011 Page 9 of 18 Page 40 of 49 www.axysanalytical.com AXYS Analytical Services Ltd. TABLE 2 PCB 86 2,2',3,4,5-Pentachlorobiphenyl PCB 87 2,2',3,4,5'-Pentachlorobiphenyl PCB 87/115/116 PCB 88 2,2',3,4,6-Pentachlorobiphenyl PCB 89 2,2',3,4,6'-Pentachlorobiphenyl PCB 90 2,2',3,4',5-Pentachlorobiphenyl PCB 91 2,2',3,4',6-Pentachlorobiphenyl PCB 92 2,2',3,5,5'-Pentachlorobiphenyl PCB 93 2,2',3,5,6-Pentachlorobiphenyl PCB 94 2,2',3,5,6'-Pentachlorobiphenyl PCB 95 2,2',3,5',6-Pentachlorobiphenyl PCB 95/93 PCB 96 2,2',3,6,6'-Pentachlorobiphenyl PCB 97 2,2',3,4',5'-Pentachlorobiphenyl PCB 97/86 PCB 98 2,2',3,4',6'-Pentachlorobiphenyl PCB 99 2,2',4,4',5-Pentachlorobiphenyl PCB 100 2,2',4,4',6-Pentachlorobiphenyl PCB 101 2,2',4,5,5'-Pentachlorobiphenyl PCB 101/90/89 PCB 102 2,2',4,5,6'-Pentachlorobiphenyl PCB 103 2,2',4,5',6-Pentachlorobiphenyl PCB 104 2,2',4,6,6'-Pentachlorobiphenyl PCB 105 2,3,3',4,4'-Pentachlorobiphenyl PCB 105/127 PCB 106 2,3,3',4,5-Pentachlorobiphenyl PCB 107 2,3,3',4',5-Pentachlorobiphenyl PCB 107/109 PCB 108 2,3,3',4,5'-Pentachlorobiphenyl PCB 109 2,3,3',4,6-Pentachlorobiphenyl PCB 110 2,3,3',4',6-Pentachlorobiphenyl PCB 111 2,3,3',5,5'-Pentachlorobiphenyl PCB 112 2,3,3',5,6-Pentachlorobiphenyl PCB 113 2,3,3',5',6-Pentachlorobiphenyl PCB 114 2,3,4,4',5-Pentachlorobiphenyl PCB 115 2,3,4,4',6-Pentachlorobiphenyl PCB 116 2,3,4,5,6-Pentachlorobiphenyl PCB 117 2,3,4',5,6-Pentachlorobiphenyl PCB 118 2,3',4,4',5-Pentachlorobiphenyl PCB 118/116 PCB 119 2,3',4,4',6-Pentachlorobiphenyl PCB 120 2,3',4,5,5'-Pentachlorobiphenyl PCB 121 2,3',4,5',6-Pentachlorobiphenyl PCB 122 2,3,3',4',5'-Pentachlorobiphenyl PCB 123 2,3',4,4',5'-Pentachlorobiphenyl PCB 124 2,3',4',5,5'-Pentachlorobiphenyl PCB 125 2,3',4',5',6-Pentachlorobiphenyl PCB 126 3,3',4,4',5-Pentachlorobiphenyl PCB 127 3,3',4,5,5'-Pentachlorobiphenyl PCB 128 2,2',3,3',4,4'-Hexachlorobiphenyl PCB 129 2,2',3,3',4,5-Hexachlorobiphenyl PCB 130 2,2',3,3',4,5'-Hexachlorobiphenyl PCB 131 2,2',3,3',4,6-Hexachlorobiphenyl PCB 131/142 Canadian Association for Laboratory Accreditation (CALA) Accreditation No.: A 2637 W S Pulp T 15 15 10 15 15 15 10, 15 15 15 15 15 10 15 15 10 15 10, 15 15 15 10 15 15 15 15 10 15 15 10 15 15 10, 15 15 15 15 10, 15 15 15 15 15 10 15 15 15 15 15 15 15 10, 15 15 10, 15 10, 15 10, 15 15 10 15 15 10 15 15 15 10, 15 15 15 15 15 10 15 15 10 15 10, 15 15 15 10 15 15 15 15 10 15 15 10 15 15 10, 15 15 15 15 10, 15 15 15 15 15 10 15 15 15 15 15 15 15 10, 15 15 10, 15 10, 15 10, 15 15 10 15 15 10 15 15 15 10, 15 15 15 15 15 10 15 15 10 15 10, 15 15 15 10 15 15 15 15 10 15 15 10 15 15 10, 15 15 15 15 10, 15 15 15 15 15 10 15 15 15 15 15 15 15 10, 15 15 10, 15 10, 15 10, 15 15 10 Washington State Department of Ecology Lab. ID: C404 NP S W 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 ACC-101 Rev 06, 29-Jun-2011 Page 10 of 18 Page 41 of 49 www.axysanalytical.com AXYS Analytical Services Ltd. TABLE 2 PCB 132 2,2',3,3',4,6'-Hexachlorobiphenyl PCB 133 2,2',3,3',5,5'-Hexachlorobiphenyl PCB 134 2,2',3,3',5,6-Hexachlorobiphenyl PCB 134/143 PCB 135 2,2',3,3',5,6'-Hexachlorobiphenyl PCB 136 2,2',3,3',6,6'-Hexachlorobiphenyl PCB 137 2,2',3,4,4',5-Hexachlorobiphenyl PCB 138 2,2',3,4,4',5'-Hexachlorobiphenyl PCB 138/163/164 PCB 139 2,2',3,4,4',6-Hexachlorobiphenyl PCB 140 2,2',3,4,4',6'-Hexachlorobiphenyl PCB 141 2,2',3,4,5,5'-Hexachlorobiphenyl PCB 142 2,2',3,4,5,6-Hexachlorobiphenyl PCB 143 2,2',3,4,5,6'-Hexachlorobiphenyl PCB 144 2,2',3,4,5',6-Hexachlorobiphenyl PCB 144/135 PCB 145 2,2',3,4,6,6'-Hexachlorobiphenyl PCB 146 2,2',3,4',5,5'-Hexachlorobiphenyl PCB 147 2,2',3,4',5,6-Hexachlorobiphenyl PCB 148 2,2',3,4',5,6'-Hexachlorobiphenyl PCB 149 2,2',3,4',5',6-Hexachlorobiphenyl PCB 149/139 PCB 150 2,2',3,4',6,6'-Hexachlorobiphenyl PCB 151 2,2',3,5,5',6-Hexachlorobiphenyl PCB 152 2,2',3,5,6,6'-Hexachlorobiphenyl PCB 153 2,2',4,4',5,5'-Hexachlorobiphenyl PCB 154 2,2',4,4',5,6'-Hexachlorobiphenyl PCB 155 2,2',4,4',6,6'-Hexachlorobiphenyl PCB 156 2,3,3',4,4',5-Hexachlorobiphenyl PCB 157 2,3,3',4,4',5'-Hexachlorobiphenyl PCB 158 2,3,3',4,4',6-Hexachlorobiphenyl PCB 158/160 PCB 159 2,3,3',4,5,5'-Hexachlorobiphenyl PCB 160 2,3,3',4,5,6-Hexachlorobiphenyl PCB 161 2,3,3',4,5',6-Hexachlorobiphenyl PCB 162 2,3,3',4',5,5'-Hexachlorobiphenyl PCB 163 2,3,3',4',5,6-Hexachlorobiphenyl PCB 164 2,3,3',4',5',6-Hexachlorobiphenyl PCB 165 2,3,3',5,5',6-Hexachlorobiphenyl PCB 166 2,3,4,4',5,6-Hexachlorobiphenyl PCB 167 2,3',4,4',5,5'-Hexachlorobiphenyl PCB 168 2,3',4,4',5',6-Hexachlorobiphenyl PCB 169 3,3',4,4',5,5'-Hexachlorobiphenyl PCB 170 2,2',3,3',4,4',5-Heptachlorobiphenyl PCB 170/190 PCB 171 2,2',3,3',4,4',6-Heptachlorobiphenyl PCB 172 2,2',3,3',4,5,5'-Heptachlorobiphenyl PCB 172/192 PCB 173 2,2',3,3',4,5,6-Heptachlorobiphenyl PCB 174 2,2',3,3',4,5,6'-Heptachlorobiphenyl PCB 174/181 PCB 175 2,2',3,3',4,5',6-Heptachlorobiphenyl PCB 176 2,2',3,3',4,6,6'-Heptachlorobiphenyl PCB 177 2,2',3,3',4,5',6'-Heptachlorobiphenyl Canadian Association for Laboratory Accreditation (CALA) Accreditation No.: A 2637 W S Pulp T 15 15 15 10 15 10, 15 10, 15 15 10 15 15 10, 15 15 15 15 10 15 10, 15 15 15 15 10 15 10, 15 15 10, 15 15 15 10, 15 10, 15 15 10 15 15 15 15 15 15 15 15 15 15 10, 15 15 10 10, 15 15 10 15 15 10 10, 15 10, 15 10, 15 15 15 15 10 15 10, 15 10, 15 15 10 15 15 10, 15 15 15 15 10 15 10, 15 15 15 15 10 15 10, 15 15 10, 15 15 15 10, 15 10, 15 15 10 15 15 15 15 15 15 15 15 15 15 10, 15 15 10 10, 15 15 10 15 15 10 10, 15 10, 15 10, 15 15 15 15 10 15 10, 15 10, 15 15 10 15 15 10, 15 15 15 15 10 15 10, 15 15 15 15 10 15 10, 15 15 10, 15 15 15 10, 15 10, 15 15 10 15 15 15 15 15 15 15 15 15 15 10, 15 15 10 10, 15 15 10 15 15 10 10, 15 10, 15 10, 15 Washington State Department of Ecology Lab. ID: C404 NP S W 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 ACC-101 Rev 06, 29-Jun-2011 Page 11 of 18 Page 42 of 49 www.axysanalytical.com AXYS Analytical Services Ltd. TABLE 2 PCB 178 2,2',3,3',5,5',6-Heptachlorobiphenyl PCB 179 2,2',3,3',5,6,6'-Heptachlorobiphenyl PCB 180 2,2',3,4,4',5,5'-Heptachlorobiphenyl PCB 181 2,2',3,4,4',5,6-Heptachlorobiphenyl PCB 182 2,2',3,4,4',5,6'-Heptachlorobiphenyl PCB 183 2,2',3,4,4',5',6-Heptachlorobiphenyl PCB 184 2,2',3,4,4',6,6'-Heptachlorobiphenyl PCB 185 2,2',3,4,5,5',6-Heptachlorobiphenyl PCB 186 2,2',3,4,5,6,6'-Heptachlorobiphenyl PCB 187 2,2',3,4',5,5',6-Heptachlorobiphenyl PCB 187/182 PCB 188 2,2',3,4',5,6,6'-Heptachlorobiphenyl PCB 189 2,3,3',4,4',5,5'-Heptachlorobiphenyl PCB 190 2,3,3',4,4',5,6-Heptachlorobiphenyl PCB 191 2,3,3',4,4',5',6-Heptachlorobiphenyl PCB 192 2,3,3',4,5,5',6-Heptachlorobiphenyl PCB 193 2,3,3',4',5,5',6-Heptachlorobiphenyl PCB 194 2,2',3,3',4,4',5,5'-Octachlorobiphenyl PCB 195 2,2',3,3',4,4',5,6-Octachlorobiphenyl PCB 196 2,2',3,3',4,4',5,6'-Octachlorobiphenyl PCB 196/203 PCB 197 2,2',3,3',4,4',6,6'-Octachlorobiphenyl PCB 198 2,2',3,3',4,5,5',6-Octachlorobiphenyl PCB 199 2,2',3,3',4,5,5',6'-Octachlorobiphenyl PCB 200 2,2',3,3',4,5,6,6'-Octachlorobiphenyl PCB 201 2,2',3,3',4,5',6,6'-Octachlorobiphenyl PCB 202 2,2',3,3',5,5',6,6'-Octachlorobiphenyl PCB 203 2,2',3,4,4',5,5',6-Octachlorobiphenyl PCB 204 2,2',3,4,4',5,6,6'-Octachlorobiphenyl PCB 205 2,3,3',4,4',5,5',6-Octachlorobiphenyl PCB 206 2,2',3,3',4,4',5,5',6-Nonachlorobiphenyl PCB 207 2,2',3,3',4,4',5,6,6'-Nonachlorobiphenyl PCB 208 2,2',3,3',4,5,5',6,6'-Nonachlorobiphenyl PCB 209 Decachlorobiphenyl Total Monochlorobiphenyls Total Dichlorobiphenyls Total Trichlorobiphenyls Total Tetrachlorobiphenyls Total Pentachlorobiphenyls Total Hexachlorobiphenyls Total Heptachlorobiphenyls Total Octachlorobiphenyls Total Nonachlorobiphenyls Total Decachlorobiphenyls Total Polychlorinated biphenyls Aroclors Aroclor 1260 Aroclor 1254 Aroclor 1268 Aroclor 1221 Aroclor 1232 Aroclor 1248 Aroclor 1016 Canadian Association for Laboratory Accreditation (CALA) Accreditation No.: A 2637 W S Pulp T 10, 15 10, 15 10, 15 15 15 10, 15 15 10, 15 15 15 10 15 10, 15 15 10, 15 15 10, 15 10, 15 10, 15 15 10 10, 15 10, 15 10, 15 15 10, 15 15 15 15 10, 15 10, 15 10, 15 10, 15 10, 15 15 10, 15 10, 15 10, 15 10, 15 10, 15 10, 15 10, 15 10, 15 10 10 10, 15 10, 15 10, 15 15 15 10, 15 15 10, 15 15 15 10 15 10, 15 15 10, 15 15 10, 15 10, 15 10, 15 15 10 10, 15 10, 15 10, 15 15 10, 15 15 15 15 10, 15 10, 15 10, 15 10, 15 10, 15 15 10, 15 10, 15 10, 15 10, 15 10, 15 10, 15 10, 15 10, 15 10 10 10, 15 10, 15 10, 15 15 15 10, 15 15 10, 15 15 15 10 15 10, 15 15 10, 15 15 10, 15 10, 15 10, 15 15 10 10, 15 10, 15 10, 15 15 10, 15 15 15 15 10, 15 10, 15 10, 15 10, 15 10, 15 15 10, 15 10, 15 10, 15 10, 15 10, 15 10, 15 10, 15 10, 15 10 10 Washington State Department of Ecology Lab. ID: C404 NP S W 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 77 7 10 10 10 10 10 10 10 10 10 10 10 10 10 7 10 7 10 10 7 10 7 10 7 7 7 7 7 7 7 7 ACC-101 Rev 06, 29-Jun-2011 Page 12 of 18 Page 43 of 49 www.axysanalytical.com AXYS Analytical Services Ltd. TABLE 2 Aroclor 1242 Aroclor 1242/1016 Pesticides 2,4'-DDD 2,4'-DDE 2,4'-DDT 4,4'-DDD 4,4'-DDE 4,4'-DDT Aldrin Alpha-HCH Beta-HCH cis-Chlordane (alpha-Chlordane) cis-Nonachlor Delta-HCH Dieldrin Endosulphan I Endosulphan II Endosulphan sulphate Endrin Endrin aldehyde Endrin ketone Gamma-HCH (Lindane) Heptachlor Heptachlor epoxide Hexachlorobenzene Hexachlorobutadiene Methoxychlor Mirex Oxychlordane Toxaphene trans-Chlordane (gamma-Chlordane) trans-Nonachlor BDE - Brominated Diphenylethers BDE 7 2,4-dibromodiphenylether BDE 8 2,4'-dibromodiphenylether BDE 10 2,6-dibromodiphenylether BDE 11 3,3'-dibromodiphenylether BDE 12 3,4-dibromodiphenylether BDE 13 3,4'-dibromodiphenylether BDE 15 4,4'-dibromodiphenylether BDE 17 2,2',4-tribromodiphenylether BDE 25 2,3',4-tribromodiphenylether BDE 28 2,4,4'-tribromodiphenylether BDE 30 2,4,6-tribromodiphenylether BDE-33 2',3,4-tribromodiphenylether BDE 35 3,3',4-tribromodiphenylether BDE 37 3,4,4'-tribromodiphenylether BDE 47 2,2',4,4'-tetrabromodiphenylether BDE 49 2,2',4,5'-tetrabromodiphenylether BDE 66 2,3',4,4'-tetrabromodiphenylether BDE 75 2,4,4',6-tetrabromodiphenylether Canadian Association for Laboratory Accreditation (CALA) Accreditation No.: A 2637 W S Pulp T 10 10 10 Washington State Department of Ecology Lab. ID: C404 NP S W 77 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10 10, 16 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 16 10, 16 10, 16 10, 16 10 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 10, 16 16 10, 16 10, 16 10, 16 10, 16 10, 16 16 10, 16 10, 16 10, 16 10 10, 16 10, 16 16 16 16 16 16 16 16 16 16 16 16 16 16 16 16 16 16 16 16 16 16 16 16 16 16 16 16 16 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 ACC-101 Rev 06, 29-Jun-2011 Page 13 of 18 Page 44 of 49 www.axysanalytical.com AXYS Analytical Services Ltd. TABLE 2 BDE 77 BDE 85 BDE 99 BDE 100 BDE 105 BDE 116 BDE 119 BDE 126 BDE 140 BDE 153 BDE 154 BDE 155 BDE 166 BDE 181 BDE-183 BDE 190 BDE 206 BDE 207 BDE 208 BDE 209 3,3',4,4'-tetrabromodiphenylether 2,2',3,4,4'-pentabromodiphenylether 2,2',4,4',5-pentabromodiphenylether 2,2',4,4',6-pentabromodiphenylether 2,3,3',4,4'-pentabromodiphenylether 2,3,4,5,6-pentabromodiphenylether 2,3',4,4',6-pentabromodiphenylether 3,3',4,4',5-pentabromodiphenylether 2,2',3,4,4',6'-hexabromodiphenylether 2,2',4,4',5,5'-hexabromodiphenylether 2,2',4,4',5',6-hexabromodiphenylether 2,2',4,4',6,6'-hexabromodiphenylether 2,3,4,4',5,6-hexabromodiphenylether 2,2',3,4,4',5,6-heptabromodiphenylether 2,2',3,4,4',5',6-heptabromodiphenylether 2,3,3',4,4',5,6-heptabromodiphenylether 2,2',3,3',4,4',5,5',6-nonabromodiphenylether 2,2',3,3',4,4',5,6,6'-nonabromodiphenylether 2,2',3,3',4,5,5',6,6'-nonabromodiphenylether Decabromodiphenylether PFC - Perfluorinated Organic Compounds Perfluorobutanoate (PFBA) Perfluoropentanoate (PFPeA) Perfluorohexanoate (PFHxA) Perfluoroheptanoate (PFHpA) Perfluorooctanoate (PFOA) Perfluorononanoate (PFNA) Perfluorodecanoate (PFDA) Perfluoroundecanoate (PFUnA) Perfluorododecanoate (PFDoA) Perfluorobutanesulfonate (PFBS) Perfluorohexanesulfonate (PFHxS) Perfluorooctanesulfonate (PFOS) Perfluorooctane sulfonamide (PFOSA) PAH Anthracene Pyrene Benzo[ghi]perylene Benzo[e]pyrene Indeno[1,2,3-cd]pyrene Perylene Benzo[b]fluoranthene Fluoranthene Benzo[k]fluoranthene Acenaphthylene Chrysene Benzo[a]pyrene Dibenz[ah]anthracene Benz[a]anthracene Acenaphthene Phenanthrene Fluorene Canadian Association for Laboratory Accreditation (CALA) Accreditation No.: A 2637 W S Pulp T 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 17 Washington State Department of Ecology Lab. ID: C404 NP S W 14 12 14 12 14 12 14 12 14 12 14 12 14 12 14 12 14 12 14 12 14 12 14 12 14 12 13 13 13 13 13 13 13 13 13 13 13 13 13 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 ACC-101 Rev 06, 29-Jun-2011 Page 14 of 18 Page 45 of 49 www.axysanalytical.com AXYS Analytical Services Ltd. TABLE 2 Naphthalene PPCP (Pharmaceutical and Personal Care Products) Acetaminophen Azithromycin Caffeine Carbadox Carbamazepine Cefotaxime Ciprofloxacin Clarithromycin Clinafloxacin Cloxacillin Dehydronifedipine Digoxigenin Digoxin Diltiazem 1,7-Dimethylxanthine Diphenhydramine Enrofloxacin Erythromycin Flumequine Fluoxetine Lincomycin Lomefloxacin Miconazole Norfloxacin Norgestimate Ofloxacin Ormetoprim Oxacillin Oxolinic acid Penicillin G Penicillin V Roxithromycin Sarafloxacin Sulfachloropyridazine Sulfadiazine Sulfadimethoxine Sulfamerazine Sulfamethazine Sulfamethizole Sulfamethoxazole Sulfanilamide Sulfathiazole Thiabendazole Trimethoprim Tylosin Virginiamycin Anhydrochlortetracycline (ACTC) Anhydrotetracycline (ATC) Chlortetracycline (CTC) Demeclocycline Canadian Association for Laboratory Accreditation (CALA) Accreditation No.: A 2637 W S Pulp T 18 18 Washington State Department of Ecology Lab. ID: C404 NP S W 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 ACC-101 Rev 06, 29-Jun-2011 Page 15 of 18 Page 46 of 49 www.axysanalytical.com AXYS Analytical Services Ltd. TABLE 2 Doxycycline 4-Epianhydrochlortetracycline (EACTC) 4-Epianhydrotetracycline (EATC) 4-Epichlortetracycline (ECTC) 4-Epioxytetracycline (EOTC) 4-Epitetracycline (ETC) Isochlortetracycline (ICTC) Minocycline Oxytetracycline (OTC) Tetracycline (TC) Bisphenol A Furosemide Gemfibrozil Glipizide Glyburide Hydrochlorothiazide 2-hydroxy-ibuprofen Ibuprofen Naproxen Triclocarban Triclosan Warfarin Albuterol Amphetamine Atenolol Atorvastatin Cimetidine Clonidine Codeine Cotinine Enalapril Hydrocodone Metformin Oxycodone Ranitidine Triamterene Alprazolam Amitriptyline Amlodipine Benzoylecgonine Benztropine Betamethasone Cocaine DEET (N,N-diethyl-m-toluamide) Desmethyldiltiazem Diazepam Fluocinonide Fluticasone propionate Hydrocortisone 10-hydroxy-amitriptyline Meprobamate Canadian Association for Laboratory Accreditation (CALA) Accreditation No.: A 2637 W S Pulp T 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 Washington State Department of Ecology Lab. ID: C404 NP S W ACC-101 Rev 06, 29-Jun-2011 Page 16 of 18 Page 47 of 49 www.axysanalytical.com TABLE 2 Methylprednisolone Metoprolol Norfluoxetine Norverapamil Paroxetine Prednisolone Prednisone Promethazine Propoxyphene Propranolol Sertraline Simvastatin Theophylline Trenbolone Trenbolone acetate Valsaran Verapamil AXYS Analytical Services Ltd. Canadian Association for Laboratory Accreditation (CALA) Accreditation No.: A 2637 W S Pulp T 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 19 Washington State Department of Ecology Lab. ID: C404 NP S W Table 1 and Table 2 - Explanation of Terms Used: NELAP = National Environmental Laboratory Accreditation Program Non-potable water = water not fit for consumption without treatment as it may contain pollutants, contaminants, minerals or infective agents. Surface water, ground water, rainwater, effluents as well as any other non-drinking water sources are included in this category. Solid = environmental solid sample. Soil, sediment, biosolids, hazardous waste, mixed phase samples with significant solids content are included in this category. Performance based implementation = methodology follows that of the method reference but modifications deemed by AXYS as minor 1may apply, results meet method reference data quality standard. Performance based modification = modifications deemed by AXYS as significant2 have been made to method reference protocol, results meet method reference accuracy standard. The suitability of the methodology for any method prescriptive applications should be assessed based on the modifications made and the specific work requirements. Performance based method = an in-house AXYS method, published method reference not applicable. GC/LRMS = gas chromatography, low resolution mass spectrometry detection. GC/HRMS = gas chromatography, high resolution mass spectrometry detection. GC/ECD = gas chromatography, electron capture detection. LC/MS-MS = liquid chromatography, mass spectrometry-mass spectrometry detection. ACC-101 Rev 06, 29-Jun-2011 Page 17 of 18 Page 48 of 49 www.axysanalytical.com AXYS Analytical Services Ltd. Note 1: Performance Based Implementation - Examples of Minor Modifications - use of additional isotopically labeled references - adjustment of calibration range - adjustment of clean-up technique - use of a different extraction of same general type (example soxhlet vs soxhlet Dean Stark) - addition of matrix type using same principles (example addition of tissue matrix using same detection principle and similar extraction type) Note 2: Performance Based Modification - Examples of Significant Modifications - different acquisition conditions using same detection principle (example MS SIM vs. full scan) - different internal control limits while meeting method reference accuracy standard ACC-101 Rev 06, 29-Jun-2011 Page 18 of 18 Page 49 of 49