Document oeN9XmX43m6yLqpo9eG1QVDpr

3 Analytical Report Determination of PFC Concentrations in Fish Collected from the Tennessee River in the Fall of 2012 Study Title Analytical Study ISO12-01-01-04: Tennessee River Fish Sample Analysis, October 2012 Project Coordinator(s) Jai Kesari, P.E., DEE and Charles Young Weston Solutions, Inc. 1400 Weston Way Westchester, MA 19380 Principal Analytical Investigator Cleston C. Lange, Ph.D. 3M Environmental Laboratory Report Completion Date Date of Final Signature Analytical Laboratory 3M Environmental Health and Safety Operations Environmental Laboratory 3M Center, Bldg 260-05-N-17 St. Paul, MN 55144 Project Identification ISO12-01-01-04 Total Number of Pages,. 39 This laboratory maintains A2LA accreditation to: ANSI/ISO/IEC 17025:2005for the specific tests/calibrations listed in A2LA Certificate # 2052.01 and meets the principles o fISO 9001:2000. The test results included in this report are covered by this accreditationfollowing method ETS-8-045.1 ISO12-01-01-04 Tennessee River Fish Study Fall 2012 Table of Contents Table of Contents........................................................................................................................................... 2 List of Tables....................................................................................................................................................3 1 Study Information.................................................................................................................................... 4 2 Sum m ary..................................................................................................................................................5 3 Introduction.............................................................................................................................................15 4 Reference Substances.......................................................................................................................... 16 5 Method Summary...................................................................................................................................19 5.1 M ethods.....................................................................................................................................19 5.2 Sample Receipt........................................................................................................................ 19 5.3 Sample Preparation/Extraction................................................................................................ 20 5.4 LC/MS/MS Analysis.................................................................................................................. 20 6 Results .................................................................................................................................................. 24 6.1 Calibration ................................................................................................................................ 24 6.2 Limits of Quantitation (LO Q s).................................................................................................. 25 6.3 Continuing Calibration.............................................................................................................. 25 6.4 Blanks........................................................................................................................................ 26 6.5 Laboratory Control Samples (LCSs)........................................................................................ 27 6.6 Laboratory Matrix Spikes (LM Ss)............................................................................................ 34 6.7 NIST SRM 1947 Results.......................................................................................................... 34 6.8 Fillet Equipment Aqueous Rinseate Results...........................................................................34 7 Conclusion............................................................................................................................................. 36 8 Data/Sample Retention......................................................................................................................... 37 9 List of Attachments................................................................................................................................ 37 10 Signatures............................................................................................................................................. 37 2 ISO12-01-01-04 Tennessee River Fish Study Fall 2012 List of Tables Table 1. Concentration Results for Tennessee River Fish Tissue Samples..............................................6 Table 2. Laboratory Matrix Spike (LMS) Recoveries................................................................................. 14 Table 3. Reference Substances: Target Analytes..................................................................................... 16 Table 4. Reference Substances: Stable Isotope Labeled Internal Standards (IS )..................................17 Table 5. Reference Substances: Stable Isotope Labeled Surrogate Recovery Standards (SR S).......19 Table 6. LC/MS/MS Instrument Information (ETS-8-045)..........................................................................21 Table 7. Gradient Liquid Chromatography Conditions (ETS-8-045); C7-C12PFCAs, PFSAs and SRSs..................................................................................................................................................... 21 Table 8. Gradient Liquid Chromatography Conditions (ETS-8-045); for C4-C6PFCAs.......................... 22 Table 9. Analytes, Surrogates, Internal Standards, and Transitions.........................................................22 Table 10. Analytical Batches....................................................................................................................... 23 Table 11. LCS Results ............................................................................................................................... 28 Table 12. Concentration Results for Fillet-Process Equipment Rinseate Blanks....................................35 Table A 1. Concentration Results for Bass & Catfish Fillet Control Materials (Right Side Fillets)........... 38 3 1 Study Information Sponsor 3M Company Project Requestor Gary Hohenstein Manager, Environmental/ Regulatory Affairs Special Projects, EHS Operations Bldg 224-5W-03 St. Paul, MN 55144 Phone: (651) 737-3570 gahohenstein@mmm.com Project Coordinator(s) Jai Kesari, P.E., DEE Charles Young Weston Solutions, Inc. 1400 Weston Way Westchester, MA 19380 Jai phone: 610-701-3761 Charles phone: 610-701-3787 Jai.kesari@westonsolutions.com Charles.young@westonsolutions.com Principal Analytical Investigator (PAI) Cleston C. Lange, Ph.D. 3M Environmental Laboratory 3M Center, Bldg 260-5N-17 St. Paul, MN 55144 Phone: (651)-733-9860 clange@mmm.com Analytical Testing Facility 3M EHS Operations 3M Environmental Laboratory Building 260-5N-17 St. Paul, MN 55144 Study Personnel (3M Environmental Laboratory) Marlene Heying, Analyst (Pace Analytical, Professional Services) Jon Steege, Analyst (Pace Analytical, Professional Services) Patrick Kenney, Analyst (Pace Analytical, Professional Services) Kevin Eich, Analyst (Quality Associates, Inc., Professional Services) Laura Harrington, Analyst (Pace Analytical, Professional Services) Study Dates Study Initiation: October 12, 2012 Analytical Initiation: December 5, 2012 Analytical Completion: February 6, 2012 Report Completion: Date of Final Signature IS 0 12-01 -01 -04 Tennessee River Fish Study Fall 2012 4 IS 0 12-01 -01 -04 Tennessee River Fish Study Fall 2012 2 Summary Reported herein are the determined concentrations of certain perfluorinated chemicals (PFCs) measured in fish collected from the Tennessee River in the Fall of 2012. These analyses were conducted as 3M Environmental analytical project ISO12-01-01-04 and was performed in support of the Weston Solutions, Inc. study plan, titled "Revised Work Plan for Fall 2012 Fish and Surface Water Sampling in the Tennessee River", and dated October 2012. During the Fall of 2012, Weston Solutions, Inc. personnel caught five largemouth bass and five catfish from each of fifteen pre-defined fish sampling reaches located within the confines of the Wilson and Wheeler Reservoirs of the Tennessee River, located near Decatur, AL. The fish were filleted (skin removed) in the field and provided blind to the laboratory in terms of specific information on fish collections. A total of 175 fish tissue samples were received and quantitatively analyzed. The PFCs analyzed were perfluorobutanoate (PFBA), perfluorohexanoate (PFHxA), perfluoroheptanoate (PFHpA), perfluorooctanoate (PFOA), perfluorobutane sulfonate (PFBS), perfluorohexanesulfonate (PFHS), perfluorooctanesulfonate (PFOS) , perfluorobutanesulfonamide (FBSA), perfluorooctanesulfonamide (FOSA),2-[Nmethyl-FOSA]acetic acid (MeFOSAA) and 2-[N-ethyl-FOSA]acetic acid (EtFOSAA) and results reported in Table 1. Each sample was extracted in duplicate for determination of the average PFC concentrations, and approximately every10th fish sample (21 in total; 12% of total fish) had two laboratory matrix spike (LMS) samples prepared from a third and fourth 0.5 gram aliquot and fortified with known quantities of target analytes. The LMS recovery results were used as a measure of the sample measurement accuracy. The average recoveries of LMSs for all analytes were within 100 +10%, except FBSA which was 82.3%. The average recovery for PFOS was 97.2% with an RSD of 8.6%. The LMS recovery results are shown in Table 2. Each sample preparation also received two surrogate recovery standards (SRSs) 13C4-PFOA and 13C4-PFOS. The recovery of 13C4-PFOS was 88.0% (RSD = 3.8%) and for 13C4-PFOA was 85.2% (RSD = 5.2%). The overall RSDs determined from the SRS recovery results for samples were used as a measure of the analytical method precision and indicated excellent precision. Laboratory control spikes (LCSs) were prepared at 3 levels, each level in triplicate, and analyzed with each analytical batch. Average LCS recoveries were within 100 + 14% at all levels, and overall, for each analyte. The LCS statistical results (for all LCS levels) were used to determine the analytical method uncertainty (95% CI) at + 18% for PFBA, + 15% for PFHxA, + 21 % for PFHpA, + 20% for PFOA, + 18% for PFBS, + 19% for PFHS, + 22% for total-PFOS, + 40% for FBSA, + 20% for FOSA, + 18% for MeFOSAA and + 19% for EtFOSAA. Overall, the sample results showed that PFOS, FBSA and FOSA were detected with highest frequencies in the Tennessee River fish fillet tissue samples, with PFOS detected in ~98% of the samples received, and FBSA and FOSA detected in ~80-90% of the samples. PFOS was measured at the highest concentration of all analytes with a maximum value of 599 ng/g (wet weight; ww) followed by FBSA with a maximum measured value of 567 ng/g (ww) and FOSA maximum measured value of 123 ng/g (ww). MeFOSAA and EtFOSAA were detected at moderate frequency of ~50%, and lower maximum levels measured at 22.0 and 19.5 ng/g, respectively. PFBS was detected at ~20% frequency in samples, with a maximum measured concentration of 3.82 ng/g (ww). The remaining analytes PFBA, PFHxA, PFHpA, PFOA, and PFHS were detected infrequently at less than 15% of samples and maximum measured values were lower than PFBS maximum value of 3.82 ng/g (ww). 5 ISO12-01-01-04 Tennessee River Fish Study Fall 2012 Table 1. Concentration Results for Tennessee River Fish Tissue Samples SAMPLE ID LIMS ID TN _11-01941 _1 S a m p le 1 P re -S tu d y H o m o g e n iz a tio n P rocess c o n tro l MSFOO1 IS O 1 2 -0 1 -0 1 -0 4 -0 0 1 IS O 1 2 -0 1 -0 1 -0 4 -0 0 2 IP F002 IS O 1 2 -0 1 -0 1 -0 4 -0 0 3 IP F003 IS O 12 -0 1 -01 -0 4 -0 0 4 M SF004 IS O 1 2 -0 1 -0 1 -0 4 -0 0 5 M SF005 IS O 12 -0 1 -01 -0 4 -0 0 6 M SF006 IS O 12 -0 1 -01 -0 4 -0 0 7 IP F 0 0 7 IS O 12 -0 1 -01 -0 4 -0 0 8 IP F009 IS O 1 2 -0 1 -0 1 -0 4 -0 0 9 M SF010 IS O 12 -0 1 -01 -0 4 -0 1 0 M SF011 IS O 1 2 -0 1 -0 1 -0 4 -0 1 1 IP F012 IS O 1 2 -0 1 -0 1 -0 4 -0 1 2 M SF013 IS O 1 2 -0 1 -0 1 -0 4 -0 1 3 IP F 0 1 4 IS O 12 -0 1 -01 -0 4 -0 1 4 M SF015 IS O 1 2 -0 1 -0 1 -0 4 -0 1 5 IP F 0 1 6 IS O 12 -0 1 -01 -0 4 -0 1 6 M SF017 IS O 12 -0 1 -01 -0 4 -0 1 7 IP F 0 1 8 IS O 12 -0 1 -01 -0 4 -0 1 8 PFBA 1.42 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 Average Concentration (ng/g; wet wt.) PFHxA PFHpA PFOA PFBS PFHS PFOS* FBSA Avg. SRS Recovery FOSA MeFOSAA EtFOSAA 13CC4PFOA 13CC4PFOS < 0 .1 0 1 < 0 .5 0 3 < 0 .9 6 0 < 0 .1 0 1 < 0.0960 0 .7 3 1 < 0 .2 5 0 [c] < 0 .2 5 3 < 0 .5 0 0 < 0 .5 0 0 107% 105% < 0 .0 5 0 3 < 0 .0 5 0 3 < 0 .0 5 0 3 < 0 .0 5 0 3 < 0 .0 5 0 3 < 0 .0 5 0 3 < 0 .0 5 0 3 < 0 .0 5 0 3 < 0 .0 5 0 3 < 0 .0 5 0 3 < 0 .0 5 0 3 < 0 .0 5 0 3 < 0 .0 5 0 3 < 0 .0 5 0 3 < 0 .0 5 0 3 < 0 .0 5 0 3 < 0 .0 5 0 3 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 0 .5 7 6 [a,c] < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 1 .1 0 < 1 .1 0 3 .8 2 < 1 .1 0 < 1 .1 0 < 1 .1 0 < 1 .1 0 < 1 .1 0 < 1 .1 0 NR < 1 .1 0 < 1 .1 0 < 1 .1 0 < 1 .1 0 < 1 .1 0 < 1 .1 0 < 1 .1 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 259 8 .4 9 18.3 167 168 173 8 .8 6 13.4 71.0 98.5 12.1 68.9 14.3 67.1 14.6 79.6 11.7 8 6 .2 [b , c] 20.2 [c] 1 6 4 [b , c] 5 8 .2 [c ] 6 0 .9 [c ] 7 9 .5 [c ] 4 .83[c] 5 .2 9 [c ] 6 8 .6 [c ] 8 1.7 [c] 9 6 .3 [b , c] 8 4.1[c] 5 2.4 [c] 56.2 [c] 55.5 [c] 51.3 [c] 2 8.6 [c] 2 .3 7 0 .9 4 7 [b] 2 .2 3 3 .0 5 3 .0 8 2 .2 6 < 0 .5 0 5 0 .487 1 .4 2 2 .0 3 4 .8 1 1 .3 7 2 .3 5 1 .2 4 2 .4 0 1 .0 9 1 .2 1 1.45 < 0 .5 0 0 1 .0 7 1 .3 7 1.35 0 .9 6 2 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 0 .8 2 9 0 .5 4 6 [a] 0 .6 5 4 [a] 0 .6 8 4 [a] 0 .6 9 6 [a] < 0 .5 0 0 < 0 .5 0 0 1 .1 9 < 0 .5 0 0 0 .714 1 .1 7 1 .1 3 0 .9 4 3 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 0 .5 1 1 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 8 9 .7 % 8 6 .1 % 8 4 .4 % 8 6 .3 % 8 5 .1 % 8 4 .4 % 8 8 .3 % 8 6 .8 % 8 3 .0 % 8 9 .0 % 8 6 .5 % 8 5 .6 % 8 4 .4 % 8 1 .8 % 8 6 .8 % 8 7 .1 % 8 5 .0 % 9 2 .6 % 8 8 .0 % 8 7 .1 % 8 7 .9 % 8 7 .1 % 8 7 .7 % 8 9 .3 % 8 8 .8 % 8 7 .4 % 9 1 .7 % 9 0 .7 % 8 7 .7 % 8 7 .5 % 8 5 .7 % 8 9 .1 % 9 2 .0 % 8 7 .9 % 6 Table 1. Concentration Results for Tennessee River Fish Tissue Samples ISO12-01-01-04 Tennessee River Fish Study Fall 2012 SAMPLE ID M SF020 M SF021 IP F022 M SF023 IP F 0 2 4 M SF025 IP F 0 2 6 M SF027 M SF028 IP F029 M SF030 IP F031 M SF033 IP F 0 3 4 M SF035 IP F 0 3 6 M SF037 M SF038 IP F039 M SF040 IP F041 M SF042 M SF043 M SF044 M SF045 LIMS ID IS O 12 -0 1 -01 -0 4 -0 2 0 IS O 1 2 -0 1 -0 1 -0 4 -0 2 1 IS O 1 2 -0 1 -0 1 -0 4 -0 2 2 IS O 1 2 -0 1 -0 1 -0 4 -0 2 3 IS O 12 -0 1 -01 -0 4 -0 2 4 IS 0 1 2 -0 1 -0 1 -0 4 -0 2 5 IS 0 1 2 -0 1 -0 1 -0 4 -0 2 6 IS 0 1 2 -0 1 -0 1 -0 4 -0 2 7 IS 0 1 2 -0 1 -0 1 -0 4 -0 2 8 IS 0 1 2 -0 1 -0 1 -0 4 -0 2 9 IS 0 1 2 -0 1 -0 1 -0 4 -0 3 0 IS 0 1 2 -0 1 -0 1 -0 4 -0 3 1 IS 0 1 2 -0 1 -0 1 -0 4 -0 3 2 IS 0 1 2 -0 1 -0 1 -0 4 -0 3 3 IS 0 1 2 -0 1 -0 1 -0 4 -0 3 4 IS 0 1 2 -0 1 -0 1 -0 4 -0 3 5 IS 0 1 2 -0 1 -0 1 -0 4 -0 3 7 IS 0 1 2 -0 1 -0 1 -0 4 -0 3 8 IS 0 1 2 -0 1 -0 1 -0 4 -0 3 9 IS 0 1 2 -0 1 -0 1 -0 4 -0 4 0 IS 0 1 2 -0 1 -0 1 -0 4 -0 4 1 IS 0 1 2 -0 1 -0 1 -0 4 -0 4 2 IS 0 1 2 -0 1 -0 1 -0 4 -0 4 3 IS 0 1 2 -0 1 -0 1 -0 4 -0 4 4 IS 0 1 2 -0 1 -0 1 -0 4 -0 4 5 PFBA < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 2 .34 [a] < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 NR < 0 .1 0 0 < 0 .1 0 0 PFHxA PFHpA < 0 .0 5 0 3 < 0 .0 5 0 3 < 0 .0 5 0 3 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 1.78 [a] < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 1.79 [a] < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 0 .1 9 8 [a] < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 0 .1 2 0 [a] < 0 .1 0 1 0 .1 1 8 [a] < 0 .1 0 1 Average Concentration (ng/g; wet wt.) Avg. SRS Recovery PFOA < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 3.13 [a] < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 1.57 [a] NR < 0 .2 4 0 < 0 .2 4 0 PFBS < 1 .1 0 < 1 .1 0 < 1 .1 0 0 .1 8 4 [b] 2 .6 6 8 0 .2 9 4 [a] 0 .8 8 7 [b] 0 .1 7 6 [b] 0 .8 5 9 0 .460 0 .590 1 .3 1 0 .4 5 9 1 .8 6 2 0 .2 1 9 0 .3 7 1 [b] 0 .5 8 2 [b] 0 .9 9 6 [b] 0 .3 0 5 [b] 0 .2 5 9 0 .4 0 5 [b] 0 .1 3 1 [b] 0 .1 3 8 [b] < 0 .2 5 2 < 0 .2 5 2 PFHS < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 1 .5 4 [a] < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 PFOS* 65.8 98.2 5 .1 0 54.9 10.3 66.4 45.5 [b] 65.3 65.9 18.6 65.2 21.9 190 8 .1 2 207 14.4 101.9 106 15.7 104 22.2 2 10 [b] 224 116 225 FBSA FOSA MeFOSAA EtFOSAA 5 6.7 [c] 23.3 [c] 36.9 [c] 3 3.0 [c] 12.3 [c] 4 7 .9 [b , c] 19.6 [c] 48.1 [c] 4 5.0 [c] 69.2 [c] 22.1 [c] 84.2 [c] 31.5 [c] 5 .38 [c] 41.3 [c] 7 .86 [c] 6 3.6 [c] 6 6.4 [c] 5 3.7 [c] 3 5.6 [c] 3 5.4 [c] 5 2.8 [c] 55.5 [c] 107 37.9 1 .1 1 0 .6 9 9 < 0 .5 0 5 0 .800 1.07 [b] 0 .887 6 .3 8 0 .988 0 .970 4 .2 0 0 .5 7 1 3 .0 1 1.12 0 .8 8 4 [b] 1 .2 8 0 .6 2 9 0 .716 0 .750 3 .5 8 1 .2 6 2 .4 1 1.25 1.32 3 .0 9 1 .9 9 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 2 .4 0 [a] < 0 .5 0 0 < 0 .5 0 0 1 .9 4 < 0 .5 0 0 0 .7 2 9 1 .6 7 < 0 .5 0 0 0 .5 9 6 < 0 .5 0 0 < 0 .5 0 0 0 .5 2 3 0 .7 1 6 0 .7 9 8 1 .3 0 2.61 [b] 2 .8 6 1 .0 6 0 .7 6 8 [b] < 0 .5 0 0 0 .5 2 0 [a] < 0 .5 0 0 0 .326 0 .2 6 0 [a] 0 .2 8 2 4 .7 3 [a] 0 .3 4 4 [b] 0 .337 0 .844 0 .3 8 8 [b] 0 .2 8 1 0 .8 4 5 < 0 .2 5 0 0 .6 0 2 < 0 .2 5 0 0 .4 2 5 0 .464 0 .3 7 3 0 .6 5 2 0 .3 8 1 1 .5 2 1 .6 2 0 .7 9 1 0 .777 13CC4PFOA 8 4 .2 % 8 5 .7 % 8 7 .6 % 8 4 .7 % 8 5 .4 % 8 5 .7 % 8 3 .4 % 9 2 .7 % 8 7 .5 % 8 7 .6 % 9 7 .3 % 8 7 .8 % 8 5 .2 % 8 5 .0 % 8 9 .0 % 8 7 .9 % 8 4 .8 % 8 8 .6 % 8 8 .0 % 8 4 .7 % 8 5 .5 % 8 2 .5 % 9 4 .1 % 8 5 .2 % 8 3 .4 % 13CC4PFOS 8 7 .5 % 8 6 .2 % 8 8 .6 % 8 5 .8 % 8 6 .1 % 8 9 .2 % 8 4 .3 % 9 2 .9 % 9 0 .2 % 8 5 .8 % 100% 8 7 .0 % 8 6 .4 % 8 5 .2 % 9 0 .2 % 8 7 .9 % 8 4 .6 % 8 8 .8 % 8 9 .6 % 8 7 .6 % 8 6 .0 % 8 2 .5 % 9 6 .0 % 9 0 .7 % 8 9 .2 % 7 Table 1. Concentration Results for Tennessee River Fish Tissue Samples ISO12-01-01-04 Tennessee River Fish Study Fall 2012 SAMPLE ID LIMS ID M SF046 IP F 0 4 7 M SF048 IP F049 M SF050 IP F051 M SF052 IP F053 IP F055 M SF056 IP F 0 5 7 M SF058 IP F059 M SF060 M SF061 M SF062 IP F063 M SF065 IP F 0 6 6 M SF067 IP F 0 6 8 IP F069 IP F 0 7 0 M SF071 M SF073 IS O 12 -0 1 -01 -0 4 -0 4 6 IS O 12 -0 1 -01 -0 4 -0 4 7 IS O 12 -0 1 -01 -0 4 -0 4 8 IS O 1 2 -0 1 -0 1 -0 4 -0 4 9 IS O 12 -0 1 -01 -0 4 -0 5 0 IS O 1 2 -0 1 -0 1 -0 4 -0 5 1 IS O 1 2 -0 1 -0 1 -0 4 -0 5 2 IS O 1 2 -0 1 -0 1 -0 4 -0 5 3 IS O 1 2 -0 1 -0 1 -0 4 -0 5 5 IS 0 1 2 -0 1 -0 1 -0 4 -0 5 6 IS 0 1 2 -0 1 -0 1 -0 4 -0 5 7 IS 0 1 2 -0 1 -0 1 -0 4 -0 5 8 IS 0 1 2 -0 1 -0 1 -0 4 -0 5 9 IS 0 1 2 -0 1 -0 1 -0 4 -0 6 0 IS 0 1 2 -0 1 -0 1 -0 4 -0 6 1 IS 0 1 2 -0 1 -0 1 -0 4 -0 6 2 IS 0 1 2 -0 1 -0 1 -0 4 -0 6 3 IS 0 1 2 -0 1 -0 1 -0 4 -0 6 4 IS 0 1 2 -0 1 -0 1 -0 4 -0 6 5 IS 0 1 2 -0 1 -0 1 -0 4 -0 6 6 IS 0 1 2 -0 1 -0 1 -0 4 -0 6 7 IS 0 1 2 -0 1 -0 1 -0 4 -0 6 8 IS 0 1 2 -0 1 -0 1 -0 4 -0 6 9 IS 0 1 2 -0 1 -0 1 -0 4 -0 7 0 IS 0 1 2 -0 1 -0 1 -0 4 -0 7 2 PFBA < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 PFHxA PFHpA < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .0 5 0 3 < 0 .0 5 0 3 < 0 .0 5 0 3 < 0 .0 5 0 3 < 0 .0 5 0 3 < 0 .0 5 0 3 < 0 .0 5 0 3 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 0 .1 7 6 [a] < 0 .1 0 1 0 .1 3 8 [a] < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 Average Concentration (ng/g; wet wt.) PFOA < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 PFBS < 0 .2 5 2 0 .327 0 .2 6 9 [a] < 0 .2 5 2 < 0 .2 5 2 0 .337 < 0 .2 5 2 1 .54 [c] < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 0 .327 < 0 .2 5 2 0 .2 8 9 < 0 .2 5 2 0 .854 < 0 .2 5 2 0 .3 5 3 [b] < 0 .2 5 2 < 0 .2 5 2 0 .7 3 9 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 PFHS < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 PFOS* 54.1 14.3 88.9 5 .5 9 81.6 8 .0 1 93.5 [b] 6 .6 8 11.0 54.8 7 .3 3 75.3 7 .5 6 64.7 75.5 103 6 .6 7 86.9 10.5 [c] 94.6 9 .6 0 5 .6 3 1.81 35.8 36.3 FBSA 56.5 63.9 129 20.6 116 28.0 71.1 15.9 31.5 19.3 22.8 69.7 26.1 74.2 85.9 9 2 .6 16.0 4 6 .8 37.5 38.6 4 0 .8 18.0 < 0 .2 5 0 0 .5 5 5 0 .5 2 9 Avg. SRS Recovery FOSA 1 .1 1 2 .7 1 2 .4 0 1 .2 9 2 .1 1 1 .7 1 2 .1 1 1 .3 1 1.37 [b] 0 .530 1 .5 8 1 .5 9 1 .6 9 2 .4 8 2 .2 1 2 .4 2 1 .4 3 1 .1 9 1 .9 5 1 .9 8 1 .4 8 0 .7 7 1 0 .1 5 3 [b] 0 .137 0 .1 6 1 MeFOSAA EtFOSAA 0 .4 3 5 0 .2 9 3 1 .0 0 < 0 .2 5 0 0 .8 8 3 0 .307 0 .4 4 8 [b] < 0 .2 5 0 0.333 [b] < 0 .2 5 0 < 0 .2 5 0 0 .404 < 0 .2 5 0 0 .3 6 5 [b] 0 .3 9 3 0 .4 1 9 0 .300 0 .3 3 1 0 .174 0 .3 1 3 0 .198 0 .1 4 2 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 0 .3 1 5 0 .1 1 6 [a] 0 .7 9 5 0 .1 4 1 [a] 0 .7 3 1 < 0 .1 0 0 0 .458 < 0 .1 0 0 0 .1 1 3 [b] 0 .2 2 6 [b] 0 .1 0 2 [a] 0 .384 < 0 .1 0 0 0 .447 0 .544 0 .457 0 .147 0 .284 0 .344 0 .4 9 3 0 .2 7 9 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 13CC4PFOA 8 3 .3 % 8 5 .3 % 8 3 .5 % 8 5 .5 % 8 7 .0 % 8 1 .4 % 8 4 .2 % 8 5 .0 % 8 4 .9 % 8 2 .1 % 8 0 .9 % 8 1 .6 % 8 1 .5 % 77.4% 8 3 .1 % 8 1 .4 % 8 1 .5 % 8 0 .8 % 9 2 .5 % 8 1 .2 % 9 1 .2 % 9 1 .3 % 8 4 .7 % 8 1 .2 % 8 4 .2 % 13CC4PFOS 8 9 .3 % 9 0 .8 % 9 0 .7 % 9 2 .6 % 8 9 .7 % 8 7 .2 % 8 7 .9 % 8 9 .6 % 8 7 .3 % 8 7 .9 % 8 4 .6 % 8 6 .4 % 8 5 .4 % 8 4 .4 % 8 7 .7 % 8 8 .1 % 8 2 .3 % 8 3 .2 % 9 6 .2 % 8 5 .7 % 9 1 .7 % 9 3 .0 % 8 5 .4 % 8 9 .8 % 8 6 .6 % 8 Table 1. Concentration Results for Tennessee River Fish Tissue Samples ISO12-01-01-04 Tennessee River Fish Study Fall 2012 SAMPLE ID M SF074 M SF075 IP F 0 7 6 IP F 0 7 7 IP F 0 7 8 IP F079 M SF080 M SF081 IP F082 IP F083 M SF084 M SF085 M SF086 M SF087 M SF089 M SF090 M SF091 M SF092 M SF094 IP F 9 5 M SF096 IP F 9 7 IP F 9 8 M SF099 IP F 1 0 0 LIMS ID PFBA IS O 1 2 -0 1 -0 1 -0 4 -0 7 3 IS O 12 -0 1 -01 -0 4 -0 7 4 IS O 1 2 -0 1 -0 1 -0 4 -0 7 5 IS O 12 -0 1 -01 -0 4 -0 7 6 IS O 12 -0 1 -01 -0 4 -0 7 7 IS O 12 -0 1 -01 -0 4 -0 7 8 IS O 1 2 -0 1 -0 1 -0 4 -0 7 9 IS O 12 -0 1 -01 -0 4 -0 8 0 IS O 1 2 -0 1 -0 1 -0 4 -0 8 1 IS O 1 2 -0 1 -0 1 -0 4 -0 8 2 IS O 1 2 -0 1 -0 1 -0 4 -0 8 3 IS O 12 -0 1 -01 -0 4 -0 8 4 IS O 1 2 -0 1 -0 1 -0 4 -0 8 5 IS O 12 -0 1 -01 -0 4 -0 8 6 IS O 12 -0 1 -01 -0 4 -0 8 7 IS O 12 -0 1 -01 -0 4 -0 8 8 IS O 12 -0 1 -01 -0 4 -0 9 0 IS O 1 2 -0 1 -0 1 -0 4 -0 9 1 IS O 1 2 -0 1 -0 1 -0 4 -0 9 2 IS O 1 2 -0 1 -0 1 -0 4 -0 9 3 IS O 12 -0 1 -01 -0 4 -0 9 4 IS O 1 2 -0 1 -0 1 -0 4 -0 9 5 IS O 12 -0 1 -01 -0 4 -0 9 6 IS O 12 -0 1 -01 -0 4 -0 9 7 IS O 12 -0 1 -01 -0 4 -0 9 8 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 0 .8 6 0 [a] < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 PFHxA PFHpA < 0 .0 5 0 3 < 0 .0 5 0 3 0 .1 3 9 [a] < 0 .0 5 0 3 < 0 .0 5 0 3 < 0 .0 5 0 3 < 0 .0 5 0 3 < 0 .0 5 0 3 < 0 .0 5 0 3 0 .1 3 7 [a] < 0 .0 5 0 3 < 0 .0 5 0 3 < 0 .0 5 0 3 < 0 .1 0 1 0 .2 6 0 [a] 0 .1 5 1 [a] 0 .1 7 4 [a] < 0 .1 0 1 0 .1 3 2 0 .0 9 6 5 [a] 0 .144 0 .1 3 7 [a] < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 0 .1 1 0 [a] < 0 .1 0 1 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 Average Concentration (ng/g; wet wt.) Avg. SRS Recovery PFOA < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 PFBS < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 PFHS < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 PFOS* 32.7 27.9 0 .768 1 .5 0 5 .1 7 0 .836 18.6 37.8 2 .9 2 3.41 [b] 44.1 32.6 49.5 49.0 40.8 104 159 434 69.0 4 .3 7 46.9 4 .3 7 2 .2 2 124 4 .6 9 FBSA 1.72 0 .648 < 0 .2 5 0 < 0 .2 5 0 3 .5 8 < 0 .2 5 0 0 .3 6 1 0 .8 0 2 0 .2 9 5 0 .5 1 2 [b] 7 .0 1 2 .7 2 6 .8 7 25.8 14.8 28.4 20.4 8 .5 5 28.1 1 .1 6 2 .9 5 1 .9 0 0 .867 6 .1 0 0 .5 7 5 FOSA MeFOSAA EtFOSAA 0 .2 1 5 0 .1 2 2 < 0 .1 0 1 < 0 .1 0 1 0 .2 7 7 [b] < 0 .1 0 1 0 .148 0 .1 3 5 [a] 0 .1 9 6 [a] 0 .1 2 3 [a] 1 .4 4 0 .580 0 .1 2 7 [a] 0 .3 7 9 1.22 [b] 1 .7 1 2 .2 4 4 .6 9 0 .6 7 2 0 .258 0 .396 0 .4 1 3 0 .2 9 9 [a] 1 .6 0 0 .656 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 0 .1 4 9 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 0 .398 0.122 [b] < 0 .1 0 0 0 .2 8 4 0 .2 8 8 [b] 0 .2 8 9 0 .478 0 .6 8 8 [b] 0 .4 4 6 < 0 .1 0 0 0 .1 7 1 0 .1 2 1 < 0 .1 0 0 0 .648 0 .1 2 2 [a] < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 0 .5 3 0 [b] < 0 .2 5 0 < 0 .2 5 0 0 .318 0 .3 2 2 [a] 0 .3 5 0 [a] 0 .464 0 .704 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 0 .4 7 8 [b] < 0 .2 5 0 13CC4PFOA 8 8 .2 % 8 3 .8 % 8 7 .7 % 8 5 .0 % 8 5 .0 % 8 0 .9 % 8 1 .2 % 8 8 .2 % 9 1 .1 % 8 2 .9 % 8 3 .8 % 8 4 .6 % 76.0% 78.4% 8 1 .1 % 8 5 .3 % 8 3 .8 % 8 2 .9 % 8 2 .7 % 8 2 .6 % 8 2 .9 % 8 0 .8 % 8 4 .4 % 8 1 .3 % 8 1 .6 % 13CC4PFOS 9 2 .5 % 8 8 .7 % 8 7 .2 % 8 9 .6 % 8 9 .0 % 8 2 .3 % 8 6 .4 % 9 3 .8 % 9 2 .5 % 8 8 .3 % 9 0 .6 % 8 8 .7 % 8 1 .9 % 8 4 .0 % 8 8 .2 % 8 9 .8 % 9 0 .1 % 8 7 .4 % 8 5 .8 % 8 6 .5 % 8 7 .0 % 8 7 .7 % 8 9 .2 % 8 6 .6 % 8 6 .8 % 9 Table 1. Concentration Results for Tennessee River Fish Tissue Samples ISO12-01-01-04 Tennessee River Fish Study Fall 2012 SAMPLE ID LIMS ID IP F101 M SF102 IP F103 IP F 1 0 4 IP F105 IP F 1 0 6 IP F 1 0 7 IP F 1 0 8 TN _11-01941 _ 1 S a m p le 2 M id -S tu d y H o m o g e n iz a tio n P rocess c o n tro l M SF109 M SF111 IP F112 IP F113 IP F 1 1 4 M SF115 IP F 1 1 6 IP F 1 1 7 IP F 1 1 8 M SF119 IP F 1 2 0 M SF121 IP F122 IS O 1 2 -0 1 -0 1 -0 4 -0 9 9 IS O 12 -0 1 -01 -0 4 -1 0 0 IS O 1 2 -0 1 -0 1 -0 4 -1 0 1 IS O 1 2 -0 1 -0 1 -0 4 -1 0 2 IS O 1 2 -0 1 -0 1 -0 4 -1 0 3 IS O 12 -0 1 -01 -0 4 -1 0 4 IS O 1 2 -0 1 -0 1 -0 4 -1 0 5 IS O 12 -0 1 -01 -0 4 -1 0 6 IS O 12 -0 1 -01 -0 4 -1 0 8 IS O 1 2 -0 1 -0 1 -0 4 -1 0 9 IS O 12 -0 1 -01 -0 4 -1 1 0 IS O 1 2 -0 1 -0 1 -0 4 -1 1 1 IS O 1 2 -0 1 -0 1 -0 4 -1 1 2 IS O 1 2 -0 1 -0 1 -0 4 -1 1 3 IS O 12 -0 1 -01 -0 4 -1 1 4 IS O 1 2 -0 1 -0 1 -0 4 -1 1 5 IS O 12 -0 1 -01 -0 4 -1 1 6 IS O 12 -0 1 -01 -0 4 -1 1 7 IS O 12 -0 1 -01 -0 4 -1 1 8 IS O 1 2 -0 1 -0 1 -0 4 -1 1 9 IS O 12 -0 1 -01 -0 4 -1 2 0 IS O 1 2 -0 1 -0 1 -0 4 -1 2 1 PFBA < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 1.42 < 1 .0 0 < 1 .0 0 < 1 .0 0 < 1 .0 0 < 1 .0 0 < 1 .0 0 < 1 .0 0 < 1 .0 0 < 1 .0 0 < 1 .0 0 < 1 .0 0 < 1 .0 0 < 1 .0 0 PFHxA PFHpA < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 Average Concentration (ng/g; wet wt.) PFOA < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 PFBS < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 PFHS < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 PFOS* 50.9 218 42.0 4 .1 5 3 .4 1 2 .4 2 7 .8 5 29.1 FBSA 3 .3 7 22.4 3 .5 6 1 .1 6 0 .6 3 3 0 .6 0 9 1.39 1.29 Avg. SRS Recovery FOSA 6 .6 8 1.37 [b] 6 .2 6 < 0 .2 5 3 0 .2 5 5 [a] 0 .2 7 1 [a] 1 .2 8 1 .0 7 MeFOSAA EtFOSAA 0 .2 3 6 [b] 1 .3 9 0 .2 4 3 0 .1 1 1 [a] < 0 .1 0 0 0 .1 0 7 [a] < 0 .1 0 0 0 .2 4 6 [b] < 0 .2 5 0 1 .2 7 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 13CC4PFOA 79.9% 8 2 .2 % 8 4 .9 % 78.2% 8 5 .9 % 8 2 .1 % 78.7% 8 2 .6 % 13CC4PFOS 8 9 .2 % 9 1 .2 % 9 1 .3 % 8 5 .0 % 8 7 .7 % 8 8 .2 % 8 5 .8 % 8 8 .0 % 0 .1 2 6 [a] < 0 .5 0 3 < 0 .2 4 0 < 0 .2 5 2 < 0 .2 5 0 0 .5 2 3 [a] < 0 .2 5 0 < 0 .2 5 3 < 0 .1 0 0 < 0 .2 5 0 8 2 .9 % 9 2 .4 % < 2 .5 2 < 2 .5 2 < 2 .5 2 < 2 .5 2 < 2 .5 2 < 2 .5 2 < 2 .5 2 < 2 .5 2 < 2 .5 2 < 2 .5 2 < 2 .5 2 < 2 .5 2 < 2 .5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 21.8 25.7 1 .7 8 1 .3 0 1 .2 4 31.2 2 .6 5 0 .897 2 .0 1 23.9 0 .9 6 1 19.9 4 .4 5 1.03 1 .5 7 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 4 .2 8 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 2 .2 6 < 0 .5 0 0 2 .0 6 < 0 .5 0 0 < 0 .1 0 1 0 .2 3 5 [b] < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 0 .1 6 2 0 .300 0 .1 4 1 < 0 .1 0 1 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 8 3 .8 % 8 4 .9 % 8 5 .8 % 8 8 .9 % 8 4 .1 % 77.0% 79.6% 8 6 .7 % 8 0 .6 % 8 2 .1 % 8 2 .0 % 8 1 .6 % 8 0 .6 % 8 6 .6 % 8 8 .2 % 8 8 .6 % 9 2 .0 % 8 8 .0 % 8 4 .1 % 8 5 .2 % 8 9 .4 % 8 5 .2 % 9 1 .8 % 8 4 .1 % 8 9 .6 % 8 3 .8 % 10 Table 1. Concentration Results for Tennessee River Fish Tissue Samples ISO12-01-01-04 Tennessee River Fish Study Fall 2012 SAMPLE ID LIMS ID PFBA PFHxA PFHpA IP F123 M SF124 M SF125 M SF127 M SF128 M SF129 M SF130 M SF131 M SF132 IP F133 IP F 1 3 4 IP F135 IP F 1 3 7 IP F 1 3 8 M SF139 M SF140 M SF141 M SF142 M SF143 M SF144 TN _11-01941 _1 S a m p le 3 P o st-S tu d y H o m o g e n iz a tio n IP F181 M SF145 IS O 1 2 -0 1 -0 1 -0 4 -1 2 2 IS O 1 2 -0 1 -0 1 -0 4 -1 2 3 IS O 12 -0 1 -01 -0 4 -1 2 4 IS O 12 -0 1 -01 -0 4 -1 2 6 IS O 12 -0 1 -01 -0 4 -1 2 7 IS O 12 -0 1 -01 -0 4 -1 2 8 IS O 1 2 -0 1 -0 1 -0 4 -1 2 9 IS O 12 -0 1 -01 -0 4 -1 3 0 IS O 1 2 -0 1 -0 1 -0 4 -1 3 1 IS O 1 2 -0 1 -0 1 -0 4 -1 3 2 IS O 1 2 -0 1 -0 1 -0 4 -1 3 3 IS O 12 -0 1 -01 -0 4 -1 3 4 IS O 1 2 -0 1 -0 1 -0 4 -1 3 5 IS O 12 -0 1 -01 -0 4 -1 3 6 IS O 12 -0 1 -01 -0 4 -1 3 7 IS O 12 -0 1 -01 -0 4 -1 3 8 IS O 1 2 -0 1 -0 1 -0 4 -1 3 9 IS O 12 -0 1 -01 -0 4 -1 4 0 IS O 1 2 -0 1 -0 1 -0 4 -1 4 1 IS O 1 2 -0 1 -0 1 -0 4 -1 4 2 IS O 12 -0 1 -01 -0 4 -1 4 4 IS O 1 2 -0 1 -0 1 -0 4 -1 4 5 IS O 12 -0 1 -01 -0 4 -1 4 6 < 1 .0 0 < 1 .0 0 < 1 .0 0 < 1 .0 0 < 1 .0 0 < 1 .0 0 < 1 .0 0 < 0 .2 5 0 [c] < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 1.42 < 0 .2 5 0 < 0 .2 5 0 < 2 .5 2 < 2 .5 2 < 2 .5 2 < 2 .5 2 < 2 .5 2 < 2 .5 2 < 2 .5 2 < 0 .1 0 1 0 .0 9 9 2 [a] 0 .4 1 7 [a] < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 0 .0 9 4 2 [a] < 0 .1 0 1 0 .136 < 0 .1 0 1 < 0 .1 0 1 0 .1 2 2 [a] 0 .1 7 0 [a] < 0 .1 0 1 < 0 .1 0 1 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 Average Concentration (ng/g; wet wt.) PFOA < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 PFBS < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 0 .444 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 PFHS < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 0 .416 < 0 .1 0 0 0 .1 2 3 < 0 .1 0 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 PFOS* 2 .1 1 73.2 40.8 [d] 408 262 250 333 178 324 22.6 3.98 [b] 4.89 [b] 1 .9 0 8 .5 5 279 281 134 45.0 143 76.9 FBSA < 0 .5 0 0 6 .4 7 6 .2 3 182 110 137 119 1 28 [c] 143 [c] 8 .8 7 [c] 1.80 [c] 3 .67 [c] 2 .50 [c] 3 .97 [c] 72.9 [c] 133 [c] 1 14 [c] 15.4 [c] 111 [c] 11.4 [c] Avg. SRS Recovery FOSA < 0 .1 0 1 0 .2 5 8 [b] 0 .2 0 0 [d] 4 .1 9 4 .0 0 2 .4 6 8 .1 8 2 .1 2 7 .2 1 0 .384 < 0 .2 5 3 < 0 .2 5 3 < 0 .2 5 3 < 0 .2 5 3 1.73 3 .7 5 1.75 0 .4 4 1 [b] 1.83 0 .2 4 3 [a] MeFOSAA EtFOSAA < 0 .1 0 0 < 0 .1 0 0 <0 .1 0 0 [d] 2 .2 2 1 .1 8 1 .6 5 5 .0 7 1 .6 9 5 .5 4 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 1 .4 3 1 .8 5 1 .0 6 < 0 .2 5 0 1 .0 2 < 0 .2 5 0 < 0 .5 0 0 < 0 .5 0 0 <0 .5 0 0 [d] 1 .7 3 1 .0 8 1 .1 9 2 .4 2 1 .4 3 2 .8 7 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 1 .4 5 1 .7 6 0 .7 0 3 < 0 .2 5 0 0 .7 5 3 < 0 .2 5 0 13CC4PFOA 8 3 .4 % 8 0 .7 % 78.8% 8 3 .6 % 75.5% 77.6% 78.7% 8 2 .7 % 9 0 .8 % 9 6 .5 % 8 7 .7 % 9 1 .2 % 8 5 .2 % 9 3 .1 % 8 9 .4 % 8 9 .3 % 8 6 .8 % 8 9 .7 % 9 2 .9 % 9 1 .5 % 13CC4PFOS 8 5 .4 % 8 7 .7 % 89.4% [d] 8 6 .6 % 8 2 .7 % 8 3 .7 % 8 7 .5 % 8 5 .4 % 8 5 .8 % 9 5 .7 % 8 4 .9 % 8 5 .9 % 8 7 .4 % 9 0 .0 % 8 7 .3 % 8 4 .5 % 8 9 .3 % 9 0 .5 % 9 2 .8 % 9 2 .6 % < 0 .2 4 0 < 0 .2 5 2 < 0 .2 5 0 < 0 .1 8 4 [a] < 0 .2 5 0 [c] < 0 .2 5 3 < 0 .2 5 0 < 0 .2 5 0 8 7 .3 % 8 4 .8 % < 0 .2 4 0 < 0 .2 4 0 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 0 < 0 .2 5 0 5 .0 0 175 < 0 .2 5 0 [c] 15.2 [c] < 0 .2 5 3 0 .348 < 0 .2 5 0 1 .1 6 < 0 .2 5 0 0 .896 8 4 .5 % 8 8 .2 % 8 2 .8 % 9 3 .6 % 11 Table 1. Concentration Results for Tennessee River Fish Tissue Samples ISO12-01-01-04 Tennessee River Fish Study Fall 2012 SAMPLE ID LIMS ID M SF146 IP F 1 4 7 IP F 1 4 8 IP F149 IP F 1 5 1 IP F152 IP F153 IP F155 M SF157 M SF158 IP F159 IP F 1 6 0 IP F161 IP F162 IP F163 IP F 1 6 4 IP F165 IP F 1 6 6 M SF167 M SF168 M SF169 M SF170 M SF172 M SF173 M SF174 IS O 12 -0 1 -01 -0 4 -1 4 7 IS O 12 -0 1 -01 -0 4 -1 4 8 IS O 1 2 -0 1 -0 1 -0 4 -1 4 9 IS O 12 -0 1 -01 -0 4 -1 5 0 IS 0 1 2 -0 1 -0 1 -0 4 -1 5 1 IS 0 1 2 -0 1 -0 1 -0 4 -1 5 2 IS 0 1 2 -0 1 -0 1 -0 4 -1 5 3 IS 0 1 2 -0 1 -0 1 -0 4 -1 5 4 IS 0 1 2 -0 1 -0 1 -0 4 -1 5 5 IS 0 1 2 -0 1 -0 1 -0 4 -1 5 6 IS 0 1 2 -0 1 -0 1 -0 4 -1 5 7 IS 0 1 2 -0 1 -0 1 -0 4 -1 5 8 IS 0 1 2 -0 1 -0 1 -0 4 -1 5 9 IS 0 1 2 -0 1 -0 1 -0 4 -1 6 0 IS 0 1 2 -0 1 -0 1 -0 4 -1 6 2 IS 0 1 2 -0 1 -0 1 -0 4 -1 6 3 IS 0 1 2 -0 1 -0 1 -0 4 -1 6 4 IS 0 1 2 -0 1 -0 1 -0 4 -1 6 5 IS 0 1 2 -0 1 -0 1 -0 4 -1 6 6 IS 0 1 2 -0 1 -0 1 -0 4 -1 6 7 IS 0 1 2 -0 1 -0 1 -0 4 -1 6 8 IS 0 1 2 -0 1 -0 1 -0 4 -1 6 9 IS 0 1 2 -0 1 -0 1 -0 4 -1 7 0 IS 0 1 2 -0 1 -0 1 -0 4 -1 7 1 IS 0 1 2 -0 1 -0 1 -0 4 -1 7 2 PFBA < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 Average Concentration (ng/g; wet wt.) PFHxA < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .1 0 1 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 0 .3 1 7 [a] < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 0 .2 9 4 [a] PFHpA < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 PFOA < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 4 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 < 0 .4 8 0 0 .6 4 3 < 0 .4 8 0 < 0 .4 8 0 < 0 .2 4 0 PFBS < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 < 0 .5 0 3 2 .2 1 < 0 .5 0 3 < 0 .5 0 3 < 0 .2 5 2 PFHS < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 0 .7 8 3 0 .3 9 1 < 0 .2 5 0 < 0 .5 0 0 PFOS* 80.3 26.9 69.6 64.2 11.7 29.2 16.3 11.9 115 145 11.2 4 .8 7 85.6 79.3 83.4 283 542 286 50.6 120 435 373 96.4 599 521 FBSA 17.7 [c] 7 2.8 [c] 65.9 [c] 45.9 [c] 49.5 106 10.1 38.6 74.7 67.3 20.4 8 .4 7 69.2 65.8 4 4 .8 37.5 34.1 33.3 12.1 20.2 67.2 [b] 567 4 5 .8 135 9 5 .0 Avg. SRS Recovery FOSA 0 .4 6 5 7 .2 3 3 .6 4 2 .4 5 1 .8 1 4 .9 2 0 .816 1 .8 3 1 .2 0 2 .2 8 0 .620 0 .490 34.1 3 1 .4 2 5 .6 63.5 [b] 123 8 8 .1 1 .2 8 4 .2 2 7 .4 4 59.9 2 .9 2 9 .7 5 5 .2 0 MeFOSAA EtFOSAA 0 .3 3 6 0 .8 0 6 0 .3 5 1 0 .2 9 9 < 0 .5 0 0 0 .7 3 1 < 0 .5 0 0 < 0 .5 0 0 0 .786 0 .974 < 0 .5 0 0 < 0 .5 0 0 3 .4 2 3 .0 9 1.85 3 .5 1 13.2 7 .9 6 < 0 .5 0 0 1.21 3 .5 5 22.5 0 .5 4 5 4 .6 7 1.62 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 0 .8 3 5 0 .847 < 0 .5 0 0 < 0 .5 0 0 2 .6 6 2 .5 0 1 .1 6 1 .0 0 4 .3 0 3 .3 2 < 0 .5 0 0 1 .5 6 3 .0 1 19.0 0 .6 3 9 3 .4 6 1 .4 3 13CC4PFOA 8 6 .5 % 8 9 .8 % 9 1 .2 % 9 3 .0 % 8 7 .7 % 8 5 .8 % 8 5 .2 % 77.2% 9 1 .7 % 8 5 .9 % 8 4 .1 % 8 3 .1 % 8 7 .3 % 79.3% 8 1 .4 % 8 1 .3 % 8 8 .6 % 8 5 .1 % 8 8 .1 % 8 1 .2 % 77.9% 8 6 .0 % 79.7% 8 9 .5 % 8 5 .7 % 13CC4PFOS 8 7 .2 % 9 0 .0 % 9 1 .4 % 9 1 .5 % 8 3 .1 % 8 7 .1 % 8 4 .6 % 8 5 .6 % 9 3 .2 % 8 7 .9 % 8 5 .7 % 8 5 .8 % 8 9 .7 % 8 0 .9 % 8 2 .1 % 8 9 .4 % 8 9 .6 % 8 7 .5 % 9 0 .6 % 8 1 .8 % 8 6 .0 % 9 1 .6 % 8 0 .6 % 8 9 .1 % 8 7 .7 % 12 Table 1. Concentration Results for Tennessee River Fish Tissue Samples ISO12-01-01-04 Tennessee River Fish Study Fall 2012 SAMPLE ID LIMS ID IP F175 IS 0 1 2 -0 1 -0 1 -0 4 -1 7 3 PFBA < 0 .2 5 0 Average Concentration (ng/g; wet wt.) PFHxA < 0 .2 5 2 PFHpA < 0 .2 5 2 PFOA < 0 .2 4 0 PFBS < 0 .2 5 2 PFHS < 0 .5 0 0 PFOS* 8 .5 7 FBSA 36.2 Avg. SRS Recovery FOSA 1 .3 6 MeFOSAA EtFOSAA < 0 .5 0 0 < 0 .5 0 0 13CC4PFOA 8 8 .2 % 13CC4PFOS 8 7 .9 % IP F 1 7 6 IS 0 1 2 -0 1 -0 1 -0 4 -1 7 4 < 0 .2 5 0 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 4 0 < 0 .2 5 2 < 0 .5 0 0 11.8 28.6 1 .5 4 < 0 .5 0 0 < 0 .5 0 0 8 6 .2 % 8 4 .9 % IP F 1 7 7 IS 0 1 2 -0 1 -0 1 -0 4 -1 7 5 < 0 .2 5 0 0 .4 2 1 < 0 .2 5 2 < 0 .2 4 0 < 0 .2 5 2 < 0 .5 0 0 12.7 29.9 1 .6 0 < 0 .5 0 0 < 0 .5 0 0 9 0 .7 % 9 0 .5 % IP F 1 7 8 IS 0 1 2 -0 1 -0 1 -0 4 -1 7 6 < 0 .2 5 0 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 4 0 0 .3 2 2 < 0 .5 0 0 10.5 51.2 1 .2 3 < 0 .5 0 0 < 0 .5 0 0 8 5 .3 % 8 8 .4 % IP F179 IS 0 1 2 -0 1 -0 1 -0 4 -1 7 7 < 0 .2 5 0 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 4 0 0 .506 < 0 .5 0 0 10.7 35.7 0 .670 < 0 .5 0 0 < 0 .5 0 0 8 8 .1 % 8 6 .5 % IP F 1 8 0 IS 0 1 2 -0 1 -0 1 -0 4 -1 7 8 < 0 .2 5 0 0 .277 < 0 .2 5 2 < 0 .2 4 0 < 0 .2 5 2 < 0 .5 0 0 7 .6 9 27.8 1 .4 3 0 .5 3 9 < 0 .5 0 0 9 3 .8 % 8 9 .8 % D AL-M SF-LS10001 D A L -IP F -L S 10002 IS 0 1 2 -0 1 -0 1 -0 4 -1 8 0 < 0 .2 5 0 IS 0 1 2 -0 1 -0 1 -0 4 -1 8 1 < 0 .2 5 0 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 5 2 < 0 .2 5 2 < 0 .5 0 0 < 0 .5 0 0 < 0 .3 6 6 3 .9 5 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 3 < 0 .2 5 3 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 9 2 .1 % 8 4 .3 % 9 1 .1 % 8 6 .1 % D AL-M SF-LS10003 IS 0 1 2 -0 1 -0 1 -0 4 -1 8 2 < 0 .2 5 0 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 4 0 < 0 .2 5 2 < 0 .5 0 0 3 .6 7 < 0 .2 5 0 < 0 .2 5 3 < 0 .5 0 0 < 0 .5 0 0 9 4 .7 % 9 1 .4 % D A L- IPF-LS10004 D A L- IPF-LS10005 IS 0 1 2 -0 1 -0 1 -0 4 -1 8 3 IS 0 1 2 -0 1 -0 1 -0 4 -1 8 4 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 4 0 < 0 .2 4 0 < 0 .2 5 2 < 0 .2 5 2 < 0 .5 0 0 < 0 .5 0 0 < 0 .3 6 6 < 0 .3 6 6 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 3 < 0 .2 5 3 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 < 0 .5 0 0 8 7 .3 % 8 7 .1 % 8 5 .8 % 8 4 .3 % D AL-M SF-LS10006 IS 0 1 2 -0 1 -0 1 -0 4 -1 8 5 < 0 .2 5 0 < 0 .2 5 2 < 0 .2 5 2 < 0 .2 4 0 < 0 .2 5 2 < 0 .5 0 0 4 .3 1 < 0 .2 5 0 < 0 .2 5 3 < 0 .5 0 0 < 0 .5 0 0 8 0 .4 % 8 5 .7 % Frequency o f D etectio n (n = 175) 2.9% 12% 4.0% 2.3% 19% 2.9% 98% 87% 82% 49% 45% 100% 100% Average Recovery NA NA NA NA NA NA NA NA NA NA NA 85.2% 88.1% Standard D eviation RSD NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA 4.4% 3.4% NA 5.2% 3.8% A n a ly tic a l m e th o d u n c e rta in ty (9 5% CI) a t + 18% fo r PFBA, + 15% fo r PFHxA, + 2 1% fo r PFHpA, + 2 0% fo r PFOA, + 18% fo r PFBS, + 19% fo r PFHS, + 22% fo r to ta l-P F O S , + 4 0% fo r FBSA, + 2 0% fo r FOSA, + 18% fo r M eF O S A A a nd + 19% fo r EtFOSAA NR; S a m p le had LM S p re p a re d a n d re c o v e ry w a s o u ts id e o f 1 0 0 + 5 0 % a nd re s u lt n o t re p o rte d * ; q u a n tifie d fr o m a lin e a r-P F O S (L-P FO S ) c a lib r a tio n f o r to ta l-P F O S (s u m o f lin e a r a n d b ra n c h e d is o m e r p e a k s ) in s a m p le s , L M S s a n d LCSs. [a ] V a lu e re p o rte d fo r s in g le re p lic a te re s u lts , d u p lic a te re s u lt w a s <LLO Q [b ] RPD > 20% [c] S a m p le had LM S p re p a re d a n d LM S re c o v e ry re s u lt w a s o u ts id e o f 1 0 0 + 30% , b u t w ith in 1 0 0 + 5 0% a nd w e re re p o rte d w ith an e x p a n d e d u n c e rta in ty o f + 50% [d ] V a lu e re p o rte d fo r s in g le re s u lt, d u p lic a te s a m p le a n a ly s is had p e a k s h ift o u t o f re te n tio n tim e w in d o w a n d n o re s u lt d e te rm in e d 13 ISO12-01-01-04 Tennessee River Fish Study Fall 2012 Table 2. Laboratory Matrix Spike (LMS) Recoveries LIM S ID S tu d y ID PFBA PFHxA PFHpA PFOA PFBS PFHS LMS A n a ly te R ecoveries PFOS FBSA FOSA M e F O S A A E tF O S A A IS O 12-01-01-04-001 IS O 12-01-01-04-002 IS O 12-01-01-04-011 IS O 12-01-01-04-022 IS O 12-01-01-04-023 IS O 12-01-01-04-032 IS O 12-01-01-04-043 T N 1 1-0 1 9 4-1 /1 S am ple 1 Pre s tu d y H om o g e niza tion Process C on tro l MSFOO1 MSF011 IPF022 MSF023 MSF033 MSF043 IS O 12-01-01-04-044 MSF044 IS O 12-01-01-04-053 IPF053 IS O 12-01-01-04-064 IS O 12-01-01-04-065 IS O 12-01-01-04-076 IS O 12-01-01-04-086 MSF065 IPF066 IPF077 MSF087 IS O 12-01-01-04-096 IS O 12-01-01-04-109 IPF098 MSF109 IS O 12-01-01-04-118 IS O 12-01-01-04-130 MSF119 MSF131 IS O 12-01-01-04-139 IS O 12-01-01-04-151 MSF141 IPF151 IS O 12-01-01-04-159 IS O 12-01-01-04-172 IPF161 MSF174 A verage Recovery S tan dard D eviation RSD n 107% 109% 101% 111% 109% 102% 111% 139% [a] 9 2 .1 % 109% 112% 108% 112% 107% NR 112% 106% 100% 106% 114% 112% 104% 8 6 .0 % 8 4 .2 % 135% [a] 125% 104% 104% 117% 108% 11% 10% 20 101% 102% 9 6 .8 % 106% 100% 122% 100% 9 6 .2 % 9 7 .0 % 8 9 .6 % 102% 101% 9 5 .6 % 78.2% 8 6 .2 % 103% 107% 9 8 .4 % 9 3 .6 % 8 9 .3 % 98.8% 9.0% 9.1% 21 124% 128% 126% 9 9 .4 % 9 1 .8 % 101% 9 2 .0 % 9 1 .8 % 8 5 .0 % 8 9 .0 % 8 6 .2 % 9 2 .0 % 8 8 .4 % 8 2 .6 % 8 4 .0 % 106% 104% 103% 9 3 .6 % 9 8 .6 % 98.4% 13% 14% 21 114% 116% 109% 115% 109% NR 94.0% 90.8% 91.7% 90.8% 91.7% 95.4% 90.6% 84.4% 91.9% 108% 97.5% 107% 108% 97.9% 101% 10% 10% 20 111% NR 9 5 .6 % 115% 104% NR 107% 8 9 .7 % 8 7 .6 % 9 3 .3 % 90.0% 9 4 .8 % 9 2 .8 % 8 6 .2 % 8 9 .8 % 113% 107% 112% 9 6 .0 % 9 8 .4 % 99.6% 9.6% 9.7% 19 106% 115% 100% 98.6% 91.8% 102% 140% [a] 91.6% 92.5% 97.2% 95.6% 96.0% 86.6% 84.0% 85.4% 103% 101% 97.2% 100% 103% 99.4% 12% 12% 21 113% [b] 97.5% [b] 103% 96.9 [b]% 105% [b] 102% [b] 94.7% [b] 86.4% 95.1% [b] 80.4% 88.7% 95.7% [b] 99.6% 97.0% [b] 82.5% [b] 97.4% [b] 96.9% [b] 106% [b] 90.7% [b] 101% [b] 97.2% 8.4% 8.6% 21 53.5% [a, b] 55.4% [a, b] 86.9% [b] 52.2% [a, b] 69.7% [a, b] 53.1% [a, b] 85.9% [b] 90.5% [b] 89.1% [b] 94.6% [b] 104% 93.8% [b] 92.2% 93.1% 79.8% 50.7% [a, b] 64.8% [a, b] 111% [b] 84.0% [b] 85.7% [b] 82.3% 22% 27% 21 9 5 .3 % 9 6 .6 % 9 7 .6 % 9 7 .7 % 9 4 .6 % 106% 109% 122% 108% 114% 115% 9 7 .2 % 9 7 .3 % 108% 113% 104% 9 8 .8 % 104% 77.0% [b] 9 9 .4 % 102% 10% 10% 21 107% 104% 111% 107% 115% 101% 111% 102% 99.0% 88.0% 100% 98.0% 106% 104% 96.6% 103% 105% 97.8% 105% 90.1% 109% 103% 6.6% 6.4% 21 V alues sh o w n are th o s e d e te rm in e d fo r th e Low -LM S (fo rtifie d a t n o m in a l 5 n g /g ), unless o th e rw is e n o te d fo r th o s e sh o w n fo r high LMS (fo rtifie d a t n o m in a l 125 n g /g ) NR; va lu e w as o u tsid e o f 100 + 50% and re s u lt n o t re p o rte d fo r th a t sa m ple and LMS re c o v e ry exclu d e d fro m s u m m a ry sta tistics [a ] R ecovery o u ts id e o f 1 00+ 30% , b u t w ith in 1 00 + 50% and in c lu d e d in s u m m a ry s ta tis tic s [b ] V a lu e sh o w n w as o b ta in e d fro m h ig h -le v e l LMS (fo rtifie d a t n o m in a l 2 5 0 n g /g ); a c c u ra te c o n c e n tra tio n s o f LMSs w e re used fo r re c o v e ry ca lcu la tio n s. 105% 98.1% 111% 97.8% 97.7% 88.1% 107% 93.6% 88.5% 88.7% 85.7% 82.4% 93.8% 86.0% 81.2% 82.8% 98.9% 88.9% 102% 99.5% 98.4% 94.1% 8.5% 9.0% 21 3C C4- PFOA 102% 87.1% 83.0% 88.9% 89.3% 84.9% 90.3% 82.9% 83.4% 84.4% 89.9% 86.1% 87.4% 85.6% 82.9% 81.7% 91.9% 94.2% 85.2% 87.6% 83.0% 87.2% 4.8% 5.5% 21 3C C4- PFOS 101% 9 3 .2 % 8 8 .6 % 9 3 .2 % 8 6 .3 % 9 3 .0 % 8 8 .0 % 8 9 .6 % 8 6 .3 % 8 9 .4 % 9 1 .2 % 8 7 .0 % 8 7 .4 % 9 1 .1 % 9 0 .5 % 79.4% 8 9 .4 % 8 9 .0 % 8 7 .4 % 9 1 .1 % 8 6 .6 % 89.5% 4.1% 4.5% 21 14 3 Introduction IS 0 12-01 -01 -04 Tennessee River Fish Study Fall 2012 A scientific investigation to determine the concentrations of select perfluorinated compounds (PFC) fish and surface water from the Tennessee River was conducted in the Fall of 2012, per the study work plan titled: "Revised W ork Plan for Fall 2012 Fish and Surface Water Sampling in the Tennessee River" Weston Solutions, Inc. (Westchester, MA), dated October 2012. The overall purpose of the study was to provide additional site-specific information on PFCs in fish tissues and surface water from within the confines of the Wilson and Wheeler Reservoirs, Tennessee River, located near Decatur, Alabama. Laboratory blinded fish tissue samples were provided for analysis by Weston Solutions, Inc. Weston Solutions, Inc. personnel were responsible for coordinating all field aspects of the study such as site parameters, fish and water collection procedures, handling and storage of collected specimens, documentation of individual fish information, shipment of whole fish for processing, filleting, homogenization at a contracted laboratory (TestAmerica-Burlington, VT) and the shipment of frozen fillets to TestAmerica, and then coordinating shipment of tissue homogenates to the 3M environmental Laboratory and Axys Laboratories for analysis of PFCs. TestAmerica personnel were responsible for homogenizing fillets and sending the tissue homogenates to the analytical laboratories for quantitative analysis of PFCs, under the direction of Weston Solutions, Inc. personnel. Prior to the commencement of the Tennessee River Fall 2012 fish and water sampling campaign, whole largemouth bass and catfish were purchased by the 3M Environmental Laboratory from Osage Catfisheries (Osage Beach, MO) as control materials. The right side fillet from each of three bass and three catfish were removed in the laboratory, the skin was removed from each fillet, and then control fillet tissue homogenate was prepared and kept at the laboratory. The remaining three bass and three catfish with only left side fillets intact were sent to the field, to have left side fillets removed as filleting process controls. The control fillet tissues kept at the laboratory were analyzed prior to the study to establish baseline levels for the field process control materials. The results from the extraction and LC/MS/MS analysis of the control tissues maintained at the laboratory are provided in Table A1 of Attachment A. With the exception of the homogenization process controls, this was a laboratory-blinded study and the true identities of the fish tissues were not provided and were only identifiable by the anonymous IDs assigned to the samples by Weston Solutions, Inc. personnel. Additionally, Weston Solutions, Inc. personnel are responsible for associating the determined PFC concentrations with the fish collection locations, fish species, age, weight, etc. and overall evaluation of control materials. This report focuses solely on analytical aspects controlled by the laboratory during the analysis of those supplied fish tissues, and describes the extraction procedures and quantitative LC/MS/MS analysis performed at the 3M Environmental Laboratory for measuring the target analyte concentrations. Additionally included in this report are the assessment of the precision and accuracy of the data based on several quality control (QC) parameters, as follows: 1) recovery of fortified target analytes in a control bluegill fish fillet homogenate matrix as laboratory control samples (LCSs) was assessed as a measure of intra-batch and inter-batch precision and accuracy; 2) recovery of fortified target analytes into fish sample matrix for ~10% of the samples (i.e. prepared for every 10thfish sample) as laboratory matrix spikes (LMSs), and providing a measure of sample-specific target analyte measurement accuracy; and 3) recovery of target analytes from each sample was indirectly determined from the recovery of the two stable isotope labeled surrogate recovery standards (SRSs), with 13C4-PFOS representative of the perfluorinated sulfonates (PFSAS) and MeFOSAA, EtFOSAA, FOSA and FBSA and 13C4-PFOA representative of the perfluorinated carboxylates (PFCAs). The SRSs were fortified into each sample, LMS and LCS sample aliquot prior to extraction. 15 4 Reference Substances IS 0 12-01 -01 -04 Tennessee River Fish Study Fall 2012 The reference substances used during the analyses are listed in Table 3, Table 4 and Table 5. Most were purchased from commercial vendors and exist as predominantly linear isomers. Four of the reference substances were 3M-supplied materials, and were as follows: PFBS, FBSA, FOSA and technical PFHS; technical grade materials each contain pre-determined percentages of linear and branched isomers. All standard solutions were prepared from neat materials and were corrected for purity during solution preparations. Several reference standards are provided as salts, and additional correction factors for salt cations (Na+, K+, NH4+) and protons (H+) were also applied, when applicable. All analyte concentrations were determined and reported for the anion form of each. Table 3. Reference Substances: Target Analytes Reference Substance Chemical Name Chemical Formula Identifier Source Expiration Date Storage Conditions Chemical Lot Number TCR Number Physical Description Purity Reference Substance Chemical Name Chemical Formula Identifier Source Expiration Date Storage Conditions Chemical Lot Number TCR Number Physical Description Purity PFBA PFHxA PFHpA Perfluorobutyric acid Perfluorohexanoic acid Perfluoroheptanoic acid C 3F 7C O O H CAS # 375-22-4 Sigm a 4 /2 0 /2 0 1 7 Frozen 11921CH TC R 07-0011 C olorless L iquid 9 7 .6 % PFBS P o ta ssiu m perfluorobutane sulfonate C 5F 11 C O O H CAS # 307-24-4 O akw ood 8 /1 8 /2 0 1 8 Frozen C150099B1 TC R -673 C olorless L iquid 9 8 .2 % PFHS Sodium perfluoro-1h e x a n e s u lfo n a te C 6F 13C O O H CAS # 375-85-9 A ldrich 1 2 /2 4 /2 0 1 8 Frozen PU /07219E U TC R -268 C lear C rystals 9 8 .0 % Br-PFOS (linear + branched isomers) P otassium perfluorooctane s u lfo n a te C 4F 9S O 3-K+ CA S # 29420-49-3 3M C 6F B S O 3'N a+ N /A W ellington L aboratories C 8F 17S O 3-K+ CA S # 2795-39-3 W ellington L aboratories 1 /1 0 /2 0 1 7 Frozen 10 1 TC R -121 W hite pow der 9 6 .7 % 3 /2 5 /2 0 1 5 Frozen LPFHxSAM 08 TCR 12-0021 Pow der >98% * 1 2 /1 /2 0 1 4 Frozen br-PFO SK 1111 TC R 12-0026 L iquid > 98% p urity **; 78.8% c o m p o sitio n is lin ear isom er, 2 1 .2 % is b ran ch ed isom ers Technical PFOA (linear + branched) T echnical A m m onium P e rflu o ro o c ta n o ate C 8H F 15O 2 CAS # 95328-99-7 W ellington L aboratories 3 /1 7 /2 0 1 4 Frozen T P F O A 0 3 11 TC R 12-0027 L iquid >97% * FBSA N onafluoro-1butanesulfonam ide C 4F 9S O 2N H 2 N /A 3M 1/31/2021 Frozen F-11954 TC R -314 Fine W hite C rystals 9 9 .3 % 16 Table 3. Reference Substances: Target Analytes IS 0 12-01 -01 -04 Tennessee River Fish Study Fall 2012 Reference Substance Chemical Name Chemical Formula Identifier FOSA P e rflu o ro o c ta n e sulfonam ide C 8F 17S O 2N H 2 CAS # 754-91-6 MeFOSAA N -m eth y lp erflu o ro -1octanesulfonam idoacetic acid C 1 1 H 6F 17N O 4S N /A EtFOSAA N -eth y lp erflu o ro -1octanesulfonam idoacetic acid C 12H 8F 17N O 4S N /A Source 3M W ellington L aboratories W ellington L aboratories Expiration Date 3 /2 7 /2 0 1 8 8 /2 6 /2 0 1 3 3 /1 4 /2 0 1 5 Storage Conditions Chemical Lot Number TCR Number Frozen N B #120067-108 TC R -280 Frozen NM eFOSAA0308 TC R 12-0044 Frozen N E tFO SA A 0312 TC R 12-0042 Physical Description W hite pow der L iquid L iquid Purity 98.3% ***; 22.2% branched isom ers >98% * >98% * * ; 100% Purity was assumed to calculate concentrations used in this study and solution concentrations were based on the concentration of the standard material provided by the commercial vendor and assume the vendor adjusted for purity. **; 78.8% Purity (78.8% linear isomer) was used to calculate concentrations of calibration standards, L-PFOS LCS results and CCVs. 100% purity was used to calculated LMS recoveries. ***; Calculated concentrations used in the study using 98.3% purity___________________________________________________________ Table 4. Reference Substances: Stable Isotope Labeled Internal Standards (IS) Reference Substance Chemical Name Chemical Formula Identifier Source Expiration Date Storage Conditions Chemical Lot Number TCR Number Physical Description Purity MPFBA P e rflu o ro -n -[1 ,2 ,3 ,4 13C 4] - b u ta n o i c a c id 13C 4 H F 7 O 2 IS fo r P F B A W ellington L aboratories 1 0 /3 /2 0 1 5 Frozen M PFC -C -100212 TC R 12-0038 L iquid >98% * MPFHxA P e r f lu o r o - n - [ 1 ,2 - 13C 2]hexanoic acid 13C 212C 4 H F ,, O 2 IS fo r P F H x A W ellington L aboratories 1 0 /3 /2 0 1 5 Frozen M PFC -C -100212 TC R 12-0038 L iquid >98% * MPFHpA P e rflu o ro -n - [ 13C 4]heptanoic acid 13C 412C 3 H F B O2 IS fo r P F H pA W ellington L aboratories 1 0 /3 /2 0 1 5 Frozen M PFC -C -100212 TC R 12-0039 L iquid >98% * M8PFOA P e r f lu o r o - n - [ 13C 8] -o c ta n o ic acid 13C 8H F15O 2 IS fo r P F O A an d M P F O A W ellington L aboratories 10/3/2015 Frozen M PFC -C -100212 TC R 12-0038 L iquid >98% * 17 Reference Substance Chemical Name Chemical Formula Identifier Source Expiration Date Storage Conditions Chemical Lot Number TCR Number Physical Description Purity Reference Substance Chemical Name Chemical Formula Identifier Source Expiration Date Storage Conditions Chemical Lot Number 18o 2-p f b s [18O2] -Ammonium Perfluorobutanesulfonate C4F9S18O2O' NH4+ IS for PFBS and FBSA RTI International 3/9/2015 Frozen 11546-107-2 TCR-964, TCR-965 Liquid N/A d3-MeFOSAA N-methyl-d3-perfluoro-1octanesulfonamidoacetic acid C1 1 D3H3F17NO4 S IS for N-MeFOSAA Wellington Laboratories 6/14/2015 Frozen d3NMeFOSAA0408 M3PFHxS Sodium perfluoro-1[13C3]hexanesulfonate 13C312C3F13SO3'Na+ IS for PFHS Wellington Laboratories 10/3/2015 Frozen MPFC-C-100212 TCR12-0038 Liquid >98% * d5-EtFOSAA N-ethyl-d5-perfluoro-1octanesulfonamidoacetic acid C12D5H3F17NO4S IS for N-EtFOSAA Wellington Laboratories 6/14/2015 Frozen d5NEtFOSAA0408 IS 0 12-01 -01 -04 Tennessee River Fish Study Fall 2012 M8PFOS M8FOSA Sodium perfluoro-1[13C8]octanesulfonate Perfluoro-1-[13C8]octanesulfonamide 13C8Fn SO3-Na+ 13C8Fn SO2NH2 IS for PFOS and MPFOS IS for FOSA Wellington Laboratories Wellington Laboratories 10/3/2015 10/3/2015 Frozen Frozen MPFC-C-100212 TCR12-0038 Liquid MPFC-C-100212 TCR12-0038 Liquid >98% * >98% * TCR Number TCR12-0045 TCR12-0043 Physical Description Liquid Liquid Purity >98% * >98% * _ ______ __________ * ; 100% Purity was assumed to calculate concentrations used in this study and solution concentrations were based on the concentration of the standard material provided by the commercial vendor and assume the vendor adjusted for purity.________________________________ 18 IS 0 12-01 -01 -04 Tennessee River Fish Study Fall 2012 Table 5. Reference Substances: Stable Isotope Labeled Surrogate Recovery Standards (SRS) Reference Substance 13C 4-P F O A 13C4-PFOS Chemical Name Chemical Formula Purpose/Use Source Expiration Date Storage Conditions Chemical Lot Number Perfluoro-n-[1,2,3,4-13C4]-octanoic acid 13C412C4HF1502 Surrogate for PFCAs Wellington Laboratories 12/7/2013 Frozen MPF0A1210 Sodium perfluoro-1-[1,2,3,4-13C4]octanesulfonate 13C412C4F17S03-Na+ Surrogate for PFSAs, MeFOSAA, EtFOSAA, FOSA and FBSA Wellington Laboratories 12/21/2014 Frozen MPFOS1211 TCR Number TCR11-0001 TCR12-0003 Physical Description Liquid Liquid Purity >98% * >98% * *; 100% purity was used to calculate concentrations used in this study and based on the concentration of the standard material provided by the commercial vendor. 5 Method Summary 5.1 Methods The re-homogenization of fish tissue samples, and the sample extractions and quantitative LC/MS/MS analyses were performed following validated 3M Environmental Laboratory method ETS-8-045.1 [Malinsky et. al. 2011, Analytics Chemica Acta 683:pp248-257]. The bulk of the fish tissue sample analyses occurred from December 27, 2012 through February 6, 2013, with the exception of the original analysis for the fillet process controls and the first homogenization process controls which occurred in November, 2012. The identities of the instruments used for collection of the LC/MS/MS data are listed in Table 6. The analytical batches used for reporting the results of the fish analyses are listed in Table 10. 5.2 Sample Receipt All samples were received frozen on dry ice and were shipped for overnight delivery from TestAmerica (Burlington, VT) to 3M by FedEx. Upon receipt at 3M, that sample was entered into the 3M Environmental Laboratory LIMS system as sample ID IS012-01-01-04-001 A single homogenization process control fish tissue sample was received from TestAmerica on November 22, 2011 with appropriate chain of custody, logged into LIMS as IS012-01-01-04001 and analyzed on December 6, 2012. The remainder of control fish tissues (9), Tennessee River fish fillet tissues (165) and aqueous fillet equipment rinseates (10) were received from TestAmerica on two dates: December 27, 2012 and January 3, 2013 and were logged into the same LIMS project as IS012-01-01-04002 through -185. Copies of all chains of custody for the samples received were retained within the study folder and electronically scanned into the 3M Environmental Laboratory LIMS system. Data for this study is associated with study IS012-01-01-04. 19 IS 0 12-01 -01 -04 Tennessee River Fish Study Fall 2012 5.3 Sample Preparation/Extraction For the frozen fish tissue homogenates received for this study, each was thawed ~30 minutes and then removed from its sample bag and re-homogenized with dry ice using a Robot-Coupe model RSI 2Y-1 stainless steel tissue homogenizer, then transferred back into the same sample baggie and left overnight in a freezer to allow the dry ice to dissipate. For preparative extraction of each analytical sample, a nominal 0.5 gram aliquot of re-homogenized fish tissue sample was accurately weighed into a centrifuge tube. Each fish sample was typically aliquoted in duplicate with one being the primary analytical sample and the second being a duplicate analytical sample. A stable isotope labeled PFC internal standard (IS) and stable isotope labeled surrogate recovery standard (SRS) mixture was added to each sample aliquot, after which 5.0 mL of acetonitrile was added. For LMSs, a target analyte mixture was also added. The sample was mixed and then placed into a freezer for greater than 1 hr to facilitate precipitation of proteins and fats. Samples were then centrifuged at greater than 10,000 x g to pellet the solids. Then, a 1.00 mL aliquot of supernatant extract was transferred to an autosampler vial with 0.010 mL formic acid to make the final analyzed extract. Typically, approximately twenty fish samples were prepared and extracted, each sample in duplicate, and analyzed as part of each analytical batch. Each analytical batch was also prepared with the following QC elements: (1) each sample and LMS aliquot received two SRSs (13C4-PFOA and 13C4-PFOS) and ISs prior to extraction, for evaluation of overall data precision for PFC carboxylates and PFC sulfonates/sulfonamides, respectively; (2) for approximately every tenth fish sample extracted, two additional aliquots were prepared as LMSs and fortified with known quantities of target analytes, ISs and SRSs; (3) a set of sixteen solvent calibration standards were prepared in acetonitrile; (4) a set of LCSs was prepared by fortifying known quantities of target analytes, ISs and SRSs into a control bluegill fillet tissue homogenate at three levels, each level in triplicate; (5) matrix blanks (method blanks) were nominal 0.5 g of control bluegill fillet matrix and prepared with and without IS and SRS and used to evaluate endogenous levels of analyte in the LCS matrix; and (6) solvent blanks were prepared with the acetonitrile used to make calibration standards and prepared with and without ISs and SRSs. 5.4 LC/MS/MS Analysis Analyses were performed following a previously validated and published LC/MS/MS method [Malinsky et. al. 2011, Analytics Chemica Acta 683:pp248-257], described in 3M Environmental Laboratory method ETS-8-045.1. The relative percent difference (RPD) of the duplicate analytical sample results was used as a measure of the sample-specific analytical precision. The determined recoveries of fortified target analytes from LMS samples were used as a measure of sample-specific and sample matrix analytical accuracy. Recovery of the SRSs (13C4PFOS and 13C4-PFOA) from each analytical sample was additionally used as a secondary measure of the target analyte accuracy for each sample. 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 6, Table 7, Table 8 and Table 9. Additionally, the analytical batches conducted for collection of LC/MS/MS data associated with this study are listed in Table10. 20 IS 0 12-01 -01 -04 Tennessee River Fish Study Fall 2012 Table 6. LC/MS/MS Instrument Information (ETS-8-045) Instrument Name(s) Analytical Method ID Liquid Chromatograph Guard column (instrument cleanup) Injection Volume Mass Spectrometer Ion Source Polarity Software Beethoven (Be), Ginger (g), Jonas (j) and MaryAnn (m), ETS-8-045.1 Agilent 1200 or 1290 HPLC System Placed between Purge Valve and Autosampler; Prism RP (2.1 x 50 mm, 5 mm) 20 mL , 40 mL or 50 mL API 5000 Triple Quadrapole or API 4500 Triple Quad. Turbolon Spray (Electrospray) Negative Ion Analyst 1.6.1 C7 and C8 PFCAs, PFSAs and SRSs Extraction column Analytical column Column Switching Valve Waters Corp. Oasis HLB Online Column (3 x 20 mm, 25mm); 30C Betasil C18 (2.1 x 100 mm, 5 mm); 30C 0 to 5 min (waste); 5 minutes to 21 minutes to analytical column C4 and C6 PFCAs Analytical column Prism RP (2.1 x 50 mm, 5 mm); 30*0 Table 7. Gradient Liquid Chromatography Conditions (ETS-8-045); C7-C12PFCAs, PFSAs and SRSs Step Total Time Flow Rate Number [m in ] [m L/m in] 0 0.00 0.400 1 3.00 0.400 2 3.50 0.400 3 13.5 0.400 4 15.5 0.400 5 16.0 0.400 6 18.0 0.400 7 18.3 0.400 8 21.0 0.400 Percent Mobile Phase 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 Mobile Phase B [ Acetonitrile] 3.0 3.0 30.0 60.0 60.0 90.0 90.0 3.0 3.0 21 IS 0 12-01 -01 -04 Tennessee River Fish Study Fall 2012 Table 8. Gradient Liquid Chromatography Conditions (ETS-8-045); for C4-C6PFCAs Step Num ber 0 1 2 3 4 5 6 Total Time [m in] 0.00 3.00 3.50 9.00 15.0 15.1 19.0 Flow Rate [m L/m in] 0.300 0.300 0.300 0.300 0.300 0.300 0.300 Percent M obile Phase A [5 m m A queous Am m onium Acetate with 0.1% A cetic A cid] 90.0 90.0 30.0 5.0 5.0 90.0 90.0 Percent M obile Phase B [ M ethanol] 10.0 10.0 70.0 95.0 95.0 10.0 10.0 Table 9. Analytes, Surrogates, Internal Standards, and Transitions Compound A nalyte Description PFBA (C4 Acid) PFHxA (C6 Acid) PFHpA (C7 Acid) Target Target Target PFOA (C8 Acid) Target PFBS (C4 Sulfonate) PFHS (C6 Sulfonate) Target Target PFOS (C8 Sulfonate) FBSA FOSA MeFOSAA E tFO S A A Target Target Target Target Target Internal Standards (IS) 22 M R M Transition(s) 213>169 313>269 3 1 3 > 119 363>319 363>169 413>369 413>219 413>169 299>80 299>99 399>80 399>99 499>80 499>99 499>130 298>78 498>78 570>83 570>169 570>219 584>83 584>169 584>219 IS 0 12-01 -01 -04 Tennessee River Fish Study Fall 2012 Table 9. Analytes, Surrogates, Internal Standards, and Transitions Compound A nalyte Description M R M Transition(s) [1,2,3,4 -13C]-PFBA IS for PFBA 217>172 [1,2 -13C2]-PFHxA IS for PFHxA and PFHpA 315>270 [1 ,2 ,3 ,4 ,5 ,6 ,7 ,8 -13C 8]-P F O A [18O2]-PFBS [13C3]-PFHS [1,2,3,4,5,6,7,8-13C8]-PFOS IS for PFOA and [1,2,3,4-13C4]-PFOA IS for PFBS and FBSA IS for PFHS IS for PFOS and [1,2,3,4-13C4]-PFOS 421>376 303>84 402>80 507>80 [1,2,3,4,5,6,7,8-13C8]-FOSA d3-MeFOSAA IS for FOSA IS for MeFOSAA d 5-E tF O S A A IS for Et FOSAA Surrogate Recovery Standards (SRS) 506>78 573>83 573>169 573>219 589>83 589>169 589>219 [1 ,2 ,3 ,4 -13C 4]-P F O A Surrogate for PFCAs 417>372 [1,2,3,4-13C4]-PFOS Surrogate for PFSAs, EtFOSAA, MeFOSAA, FOSA & FBSA 503>80 Multiple MS/MS ion transitions are summed when more than one transition is listed for the analyte. Table 10. Analytical Batches A n a ly tic a l Batch # 1 2 3 4 5 6 7 8 9 10 11 A n a ly tic a l Batch ID j121206a j130118a j130121a j130123a g130124a j130125a j130128a g130128a m130204a g130204a m121206a A nalytes R e p o rte d IaI PFBA, PFHxA PFBA, PFHxA PFBA, PFHxA PFBA, PFHxA PFBA, PFHxA PFBA, PFHxA PFBA, PFHxA PFBA, PFHxA PFBA, PFHxA PFBA, PFHxA PFHpA, PFOA, PFBS, PFHS, PFOS, FBSA, FOSA, MeFOSAA, EtFOSAA, 13C4-PFOA and 13C4-PFOS Samples Analyzed ISO12-01-01-04-001 ISO12-01-01-04-002 to -022 ISO12-01-01-04-023 to -043 ISO12-01-01-04-044 to -064 ISO12-01-01-04-065 to -085 ISO12-01-01-04-086 to -108 ISO12-01-01-04-109 to -129 ISO12-01-01-04-130 to -150 ISO12-01-01-04-151 to -171 ISO12-01-01-04-172 to -185 ISO12-01-01-04-001 23 Table 10. Analytical Batches IS 0 12-01 -01 -04 Tennessee River Fish Study Fall 2012 A n a ly tic a l Batch # A n a ly tic a l Batch ID A nalytes R e p o rte d IaI Samples Analyzed 12 m130122b PFHpA, PFOA, PFBS, PFHS, PFOS, FBSA, FOSA, ISO12-01-01-04-002 to -022 MeFOSAA, EtFOSAA, 13C4-PFOA and 13C4-PFOS 13 m130129b PFHpA, PFOA, PFBS, PFHS, PFOS, FBSA, FOSA, ISO12-01-01-04-023 to -043 MeFOSAA, EtFOSAA, 13C4-PFOA and 13C4-PFOS 14 Be130121b PFHpA, PFOA, PFBS, PFHS, PFOS, FBSA, FOSA, ISO12-01-01-04-044 to -064 MeFOSAA, EtFOSAA, 13C4-PFOA and 13C4-PFOS 15 Be130123b PFHpA, PFOA, PFBS, PFHS, PFOS, FBSA, FOSA, ISO12-01-01-04-065 to -085 MeFOSAA, EtFOSAA, 13C4-PFOA and 13C4-PFOS 16 Be130128a PFHpA, PFOA, PFBS, PFHS, PFOS, FBSA, FOSA, ISO12-01-01-04-086 to -108 MeFOSAA, EtFOSAA, 13C4-PFOA and 13C4-PFOS 17 Be130130a PFHpA, PFOA, PFBS, PFHS, PFOS, FBSA, FOSA, ISO12-01-01-04-109 to -129 MeFOSAA, EtFOSAA, 13C4-PFOA and 13C4-PFOS 18 j130201a PFHpA, PFOA, PFBS, PFHS, PFOS, FBSA, FOSA, ISO12-01-01-04-130 to -150 MeFOSAA, EtFOSAA, 13C4-PFOA and 13C4-PFOS 19 g130130b PFHpA, PFOA, PFBS, PFHS, PFOS, FBSA, FOSA, ISO12-01-01-04-151 to -171 MeFOSAA, EtFOSAA, 13C4-PFOA and 13C4-PFOS 20 g130201a PFHpA, PFOA, PFBS, PFHS, PFOS, FBSA, FOSA, ISO12-01-01-04-172 to -185 MeFOSAA, EtFOSAA, 13C4-PFOA and 13C4-PFOS [a] Each extract was analyzed in one analytical run for short chain acids (PFBA and PFHxA) and then re-analyzed in a second analytical run fo r PFHpA, PFOA, PFBS, PFHS, PFOS, FBSA, FOSA, MeFOSAA and EtFOSAA 6 Results All reported concentration results and recovery values are rounded to three significant figures. Statistical values (RSDs, RPDs, etc.) are rounded to two significant figures. All results were calculated using full precision raw data during data calculations in Microsoft ExcelTM, and as a result some values may vary slightly when the rounded values reported herein are carried through the same calculations. 6.1 Calibration Calibration standards were prepared by spiking varying known quantities of target analytes, SRSs and ISs into acetonitrile. A set of 16 acetonitrile solvent calibration standards were prepared with each analytical batch, ranging from nominal 0.00250 ng/mL to 250 ng/mL for target analytes and SRSs; equivalent to nominal 0.0250 ng/g to 2500 ng/g in fish tissue after adjustment for dilution during extraction. The same amount of ISs were added as were added to each fish sample during extraction. Following the analysis of 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 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. High and/or low calibration standards were excluded as needed to meet 24 IS 0 12-01 -01 -04 Tennessee River Fish Study Fall 2012 LLOQ requirements and to focus quantitation at high end or low end of the curve, as needed based on sample concentrations in the batch. For PFOS, the calibration standards were prepared with brPFOS, which is composed of a well characterized isomer mixture composed of 78.8% (w/w) linear PFOS isomer and 21.2% (w/w) of branched isomers. Because fish samples showed a predominance of linear isomer, PFOS quantitation was based on the chromatographically distinct L-PFOS peak in calibration standards and those results used to construct a L-PFOS calibration curve. Samples, LMSs and LCSs were quantified by integration of total PFOS (linear + branched isomers) using the LPFOS calibration. For LCSs, both L-PFOS and total-PFOS (branched + linear isomers) recoveries were determined and are reported in Table 11 of section 6.5. 6.2 Limits of Quantitation (LOQs) The lower limit of quantitation (LLOQ) was typically the lowest non-zero calibration standard in the curve that met linearity and accuracy requirements (100 + 30%) and for which the peak area counts were at least 2X that of the average analyte peak area counts determined from the acetonitrile solvent blanks. The LLOQ for all analytes in this study was the typically nominal 0.250 ng/g fish concentration, equivalent to the 0.0250 ng/mL standard, with some exceptions as noted in the raw data. Results in Table 1 indicate the nominal LLOQ values when results were below the LLOQ. The ULOQ was typically the nominal 250 ng/mL standard (equivalent to 2500 ng/g in fish tissue), with some exceptions as noted in the raw data. 6.3 Continuing Calibration During the course of analysis of each analytical batch, continuing calibration verifications (CCVs) were performed by regular injection of mid-level calibration standards throughout the analysis. The back calculated concentration results of CCVs were used to verify that the instrument response and the initial calibration curve were still in control. All CCVs for all analytes were within 100+ 25% and met method criteria, with the following exceptions: PFHpA:m130122b052 (993%), m130122b098 (NA, outside of curve range), m130122b110 (497%), m130122b122 (72.1%), m130122b128 (NA, outside of curve range). PFOA: m130122b052 (559%), m130122b110 (383%), m130122b122 (134%) and m130122b128 (NA, outside of curve range). PFBS: m130122b052 (425%), m130122b110 (126%), m130122b122 (2.02%) and m130131a005 (130%). PFHS:m130122b052 (NA, outside of curve range) and m130122b128 (57.6%) PFOSm130122b052 (529%), m130122b110 (4050%), j130201a074 (74.3%) and j130201a112 (71.4%) FBSA: m130122b052 (402%), m130122b110 (304%), m130122b128 (61.6%), m130129b052 (70.5%), m130131a005 (70.5%), j130201a041 (134%), j130201a074 (128%) and j130201a112 (126%). FOSA: m130122b052 (392%) and m130122b110 (31.7%). MeFOSAA: m130122b052 (160%) and m130122b110 (48.5%) EtFOSAA: m130122b052 (38.8%) and m130122b110 (18.5%) 13C4-PFOA (SRS): m130122b052 (32.3%), m130122b098 (4.06%), m130122b110 (41.6%) and m130122b128 (11.4%) 13C4-PFOS (SRS): m130122b052 (16.2%), m130122b098 (4.06%), m130122b110 (37.0%) and m130122b128 (4.07%) 25 IS 0 12-01 -01 -04 Tennessee River Fish Study Fall 2012 When CCVs failed to meet criteria, the analytical run was evaluated more closely for LMS recoveries and LCS results, and a potential cause of CCV recovery failures was evaluated and documented with the batch paperwork. For example, in analytical run m130122b the standard solution used for several CCV injections ran low and were not effectively injected, resulting in CCV values well outside of the 100 + 25% criterion. In all instances, LMS results and LCS results were additionally evaluated to provide justification for reporting the results from those batches. All CCV failures were documented into a method deviation and included in the raw data. 6.4 Blanks Three types of blanks were analyzed with fish samples: (1) acetonitrile solvent blanks were typically analyzed before or after calibration standards and CCVs for evaluating and abating any potential carry over and at times are used in conjunction with method blanks to help assign LLOQs; (2) fish matrix blanks were prepared with nominal 0.5 g of a control bluegill fillet homogenate TN11-0194-1/1 and were spiked with and without ISs and SRSs and used to provide endogenous background levels for the fish matrix used to prepare LCSs; and (3) process control blanks used to evaluate the homogenization process at TestAmerica (control sunfish tissue TN11-0194-1/1) and the process controls for the field filleting process (Bass,TN12-0365-1/13, -2/13 and -3/13 and Catfish TN12-0365-1/13, -2/13 and -3/13). The fillet process controls were 3 largemouth whole bass (TN12-0364-1/13, -2/13 and -3/13) and 3 whole catfish (TN12-0365-1/13, -2/13 and -3/13) for which the right side fillets were removed at the 3M Environmental Laboratory, homogenized and analyzed prior to fish sample receipt for the study. Those fish, with right side fillets removed, were sent to the field and left side fillets removed in the field during fish filleting process. Those process control samples were returned blinded to the laboratory regarding their true identities. Homogenization process controls were returned and were identifiable to the laboratory based on identities placed on them by TestAmerica. Final results and fish identities for process controls will be verified in the Weston Solutions, Inc. final study report. All blank results were carefully evaluated for issues such as background instrument interferences, analytical carry over during the analysis to provide a background level with which to use for assigning LLOQs, and to determine endogenous values of each analyte in the fish matrix used for preparation of LCSs. 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 sunfish control matrix TN11-0194-1/1 was the matrix used for LCS preparations, and was the same matrix used for the independent evaluation of the homogenization process (homogenization process controls) at TestAmerica. That matrix has low measurable levels of endogenous total PFOS, and other analytes, and those values subtracted from LCS results during recovery calculations. The results of all samples, including homogenization and filleting process controls are included in Table 1. 26 6.5 Laboratory Control Samples (LCSs) IS 0 12-01 -01 -04 Tennessee River Fish Study Fall 2012 Laboratory control samples (LCSs) were prepared for each analytical batch by fortifying known quantities of the target analytes, ISs and SRSs into accurately weighed (nominal 0.5 gram) aliquots of a control bluegill fillet tissue prior to extraction then carrying the LCS through the extraction and analysis procedure. The LCS sets were prepared at three levels of nominal 1.0, 10 and 100 ng/g (ww), and each level was prepared in triplicate. The PFOS used to fortify LCSs was brPFOS purchased from Wellington Laboratories and with a known 78.8% (w/w) linear isomer composition and 22.2 %(w/w) branched isomer composition. Because calibration was performed using only the chromatographically separated L-PFOS isomer peak, both LPFOS and total-PFOS recoveries from LCSs were determined. Total-PFOS was always determined from samples, LMS s and matrix blanks. The LCS results were used to determine the analytical method accuracy (average recovery) and precision (RSD). The LCS accuracy and precision results, and the analytical method uncertainties at the 95% confidence interval (95% CI; 2-times standard deviation), determined from the statistical evaluation of LCS results are shown in Table 11. The method acceptance criteria of 100 30% was typically met, with some exceptions as noted in the Table. The average low-LCS, mid-LCSs and high-LCSs recoveries over 10 analytical batches, and overall for all LCSs levels, was within 100 + 14% for all analytes and SRSs. The LCS recoveries were calculated using the following equation: TLC_S_ R,, ecovery = -(-D--e-t-e-r-m---i-n-e-d---C--o-n--c-e-n--t-r-a-t-i-o-n--o--f--L--C--S----E--n--d-o--g-e--n-o--u-s--M---a-t-r-i-x--C--o-n--c-.--)*.1,00% Spike Concentration 27 Table 11. LCS Results A nalytical B atch ID LCS ID LCS L o w -1 ra LO O rsi LCS L o w -2 rsi LCS L o w -3 a cra < " LCS M id - 1 X X CL LL LCS M id - 2 CL < C LL LCS M id - 3 CL ra LO O LCS H ig h -1 rsi rsi LCS H ig h -2 LCS H ig h -3 LCS L o w -1 _Q rsi rsi LCS L o w -2 o m LCS L o w -3 a cra LCS M id - 1 < X X CL LL LCS M id - 2 CL < C LCS M id - 3 LL CL ra 00 LCS H ig h -1 o m LCS H ig h -2 LCS H ig h -3 PFBA PFHx A PFHpA PFOA 115% 101% 105% 111% 111% 106% 109% 103% 108% 106% 107% 103% 105% 111% 103% NA 108% 102% 100% 110% 109% 108% 108% 104% 106% 9 6 .5 % 9 6 .5 % 9 3 .8 % 104% 105% 9 8 .8 % NA 84.4% 69.9% 72.5% 105% 107% 106% 87.5% 89.6% 88.7% 126% 125% 119% 106% 112% 105% NA NA NA NA NA NA NA 74.7% 63.9% 66.3% 110% 115% 111% 102% 100% 101% 109% 116% 110% 104% 106% 101% NA NA NA PFBS 108% 99.2% 104% 109% 110% 107% NA NA NA 104% 104% 98.6% 97.9% 101% 96.8% 83.6% 70.0% 6 6 .8 % [a] ISO12-01-01-04 Tennessee River Fish Study Fall 2012 A n a ly te R ecovery (% ) PFHS T o talP FO S [b] L-PFO S [b] FBSA FOSA M eFO SA A E tF O S A A 13C 4-P F O A 13C 4 -P F O S 106% 9 7 .1 % 9 8 .9 % 105% 105% 102% 103% 9 8 .6 % 101% 9 9 .3 % 102% 9 6 .8 % 9 9 .0 % 102% 9 6 .2 % 8 5 .9 % 78.0% 79.1% 118% 111% 117% 114% 116% 113% 115% 110% 112% 110% 117% 110% 111% 116% 110% NA NA NA 105% 9 6 .6 % 105% 111% 112% 110% 108% 104% 106% 108% 115% 108% 105% 111% 105% 6 7 .5 % [a] 6 7 .7 % [a] 6 9 .3 % [a] 123% 117% 120% 133% 135% 132% 126% 117% 123% 90.3% 91.8% 87.6% 78.1% 80.3% 76.6% 67.0% 9 4 .6 % 9 0 .6 % 9 1 .1 % 9 3 .4 % 9 4 .7 % 9 1 .3 % 9 6 .1 % 9 1 .2 % 9 5 .7 % 9 0 .6 % 9 5 .8 % 89.5% 9 6 .2 % 104% 99.2% NA 51.7% NA 4 9 .1 % NA 119% 102% 103% 110% 110% 103% 108% 105% 108% 9 8 .4 % 103% 9 8 .7 % 9 9 .7 % 108% 9 9 .0 % 8 7 .1 % 8 6 .1 % 8 9 .1 % 115% 100% 101% 104% 108% 104% 107% 103% 104% 9 2 .1 % 9 7 .6 % 9 2 .7 % 9 1 .8 % 100% 9 3 .8 % 8 3 .3 % 79.0% 8 4 .2 % 112% 100% 102% 106% 110% 108% 104% 104% 102% 8 7 .4 % 8 9 .6 % 8 6 .9 % 8 5 .5 % 8 7 .2 % 8 7 .7 % 8 3 .3 % 8 2 .8 % 8 7 .2 % 109% 101% 101% 104% 106% 104% 102% 101% 100% 8 8 .6 % 9 0 .8 % 8 8 .2 % 8 9 .6 % 8 8 .4 % 9 3 .3 % 8 8 .0 % 8 8 .9 % 9 2 .7 % 28 Table 11. LCS Results A nalytical B atch ID LCS ID LCS L o w -1 _Q cn rsi LCS L o w -2 o m LCS L o w -3 a cra <X" LCS M id - 1 X CLLL CL LCS M id - 2 < C LL LCS M id - 3 CL ra rsi LCS H ig h -1 o m LCS H ig h -2 LCS H ig h -3 LCS L o w -1 _a rsi LCS L o w -2 o m ai C LCS L o w -3 a cra LCS M id - 1 <" X X CL LL LCS M id - 2 CL <C LL LCS M id - 3 CL ra m rsi LCS H ig h -1 o m LCS H ig h -2 LCS H ig h -3 PFBA PFHx A PFHpA PFOA 106% 9 7 .5 % 92.3% 102% 9 4 .7 % 87.9% 105% 9 7 .4 % 92.5% 105% 101% 84.3% 103% 100% 82.7% 101% 9 9 .8 % 84.9% NA NA NA NA NA NA NA NA NA 105% 9 6 .0 % 82.3% 105% 9 5 .4 % 88.4% 103% 9 5 .4 % 84.4% 109% 107% 97.7% 109% 104% 95.9% 98.2% 9 4 .5 % 83.5% NA NA 86.1% NA NA 83.2% NA NA 87.7% 106% 104% 107% 104% 99.7% 98.8% 100% 98.1% 97.5% 95.1% 93.6% 95.3% 101% 97.1% 88.5% 83.9% 79.3% 83.8% ISO12-01-01-04 Tennessee River Fish Study Fall 2012 PFBS 105% 101% 104% 107% 107% 104% 90.3% 94.7% 89.7% 87.3% 90.4% 91.2% 93.8% 96.7% 85.2% 87.6% 87.4% 90.5% A n a ly te R ecovery (% ) PFHS T o talP FO S [b] L-PFO S [b] FBSA FOSA M eFO SA A E tF O S A A 13C 4-P F O A 13C 4 -P F O S 8 5 .0 % 8 1 .2 % 8 3 .9 % 9 0 .2 % 9 0 .2 % 8 7 .3 % 8 0 .7 % 71.6% 6 9 .4 % [a] 9 3 .6 % 8 9 .2 % 9 1 .4 % 103% 102% 9 1 .4 % 8 3 .7 % 8 1 .3 % 8 3 .7 % 101% 95.6% 102% 100% 97.4% 97.8% 115% 104% 103% 89.6% 87.1% 88.0% 98.1% 94.3% 84.9% 97.6% 97.6% 101% 9 9 .2 % 9 4 .0 % 9 9 .8 % 9 7 .4 % 9 4 .3 % 9 4 .9 % 76.0% 70.6% 6 9 .2 % [a] 8 9 .2 % 8 6 .8 % 8 8 .0 % 9 8 .1 % 9 4 .2 % 8 5 .3 % 9 0 .0 % 8 9 .0 % 9 2 .3 % 75.4% 9 3 .4 % 72.5% 9 0 .2 % 71.3% 68.8% [a] 67.4% [a] 68.4% [a] 75.4% 61.6% [a] 51.1% [a] 128% 9 3 .6 % 9 5 .9 % 9 4 .8 % 9 5 .0 % NA NA NA 104% 1 3 4 % [a] 106% 129% 107% 119% 108% 119% 113% 110% 65.7% [a] 66.6% [a] 67.6% [a] 104% 8 6 .5 % 9 4 .0 % 9 3 .6 % 9 9 .3 % 9 3 .9 % 103% 9 7 .6 % 9 5 .5 % 9 6 .8 % 8 9 .5 % 8 6 .1 % 8 4 .2 % 9 7 .0 % 9 8 .6 % 9 9 .6 % 101% 101% 9 3 .5 % 9 0 .3 % 8 2 .8 % 8 8 .8 % 9 3 .4 % 9 0 .8 % 9 2 .2 % 9 0 .1 % 8 6 .3 % 8 7 .4 % 8 2 .0 % 8 0 .4 % 8 1 .8 % 8 8 .6 % 9 3 .7 % 8 9 .9 % 9 7 .2 % 9 2 .7 % 8 5 .5 % 79.5% 78.2% 78.8% 8 6 .8 % 8 4 .2 % 8 7 .3 % 8 7 .9 % 8 6 .5 % 105% 8 4 .7 % 8 6 .0 % 8 6 .4 % 8 6 .4 % 8 7 .8 % 8 5 .6 % 9 2 .8 % 8 6 .9 % 8 1 .7 % 8 5 .1 % 8 0 .8 % 8 7 .0 % 8 9 .8 % 8 2 .9 % 8 9 .2 % 8 5 .7 % 8 6 .0 % 104% 8 9 .7 % 8 8 .3 % 8 6 .5 % 8 8 .0 % 9 0 .5 % 8 4 .1 % 9 7 .5 % 8 9 .3 % 8 5 .8 % 8 9 .1 % 8 7 .5 % 9 1 .7 % 29 Table 11. LCS Results A nalytical B atch ID LCS ID LCS L o w -1 _Q m CM LCS L o w -2 O m CL) C LCS L o w -3 a cra LCS M id - 1 <" X X CL LL LCS M id - 2 CL < C LL LCS M id - 3 CL ra rsi LCS H ig h -1 o m bo LCS H ig h -2 LCS H ig h -3 LCS L o w -1 ra 00 rsi LCS L o w -2 o m ai C LCS L o w -3 a cra LCS M id - 1 <" X X CL LL LCS M id - 2 CL < C LL LCS M id - 3 CrLa LD rsi LCS H ig h -1 o m LCS H ig h -2 LCS H ig h -3 PFBA PFHx A PFHpA PFOA 122% 104% 89.7% 116% 100% 87.6% 117% 9 6 .8 % 86.5% 106% 8 9 .5 % 84.3% 118% 9 0 .8 % 87.8% 116% 9 1 .1 % 90.6% NA NA 87.0% NA NA 85.7% NA NA 86.0% 110% 100% 89.7% 105% 9 7 .1 % 91.4% 107% 9 9 .6 % 89.0% 98.6% 9 5 .4 % 85.5% 97.2% 9 4 .3 % 80.8% 88.8% 8 5 .8 % 75.1% NA NA 86.9% NA NA 85.2% NA NA 82.8% 95.0% 93.5% 95.2% 101% 97.1% 88.5% 83.9% 79.3% 83.9% 96.0% 92.1% 96.7% 92.1% 91.1% 82.1% 85.9% 83.5% 84.1% ISO12-01-01-04 Tennessee River Fish Study Fall 2012 PFBS 93.4% 91.2% 93.7% 83.5% 85.7% 88.1% 92.3% 84.4% 87.0% 94.7% 91.4% 96.3% 90.5% 89.5% 80.9% 85.4% 81.8% 83.2% A n a ly te R ecovery (% ) PFHS T o talP FO S [b] L-PFO S [b] FBSA FOSA M eFO SA A E tF O S A A 13C 4-P F O A 13C 4 -P F O S 106% 104% 103% 8 7 .0 % 9 6 .4 % 9 5 .2 % 6 9 .2 % [a] 6 8 .5 % [a] 6 8 .1 % [a] 9 7 .4 % 9 2 .0 % 9 6 .5 % 8 9 .2 % 8 7 .4 % 8 1 .1 % 8 4 .1 % 8 0 .1 % 79.1% 95.2% 94.6% 94.8% 84.5% 94.3% 92.1% 105% 102% 103% 101% 91.7% 93.7% 87.5% 89.2% 78.9% 105% 102% 101% 9 4 .8 % 9 4 .1 % 9 3 .9 % 8 3 .9 % 9 3 .5 % 9 2 .5 % 9 3 .8 % 9 1 .6 % 9 1 .6 % 9 9 .4 % 9 0 .8 % 9 2 .6 % 8 6 .0 % 8 8 .2 % 78.0% 9 2 .2 % 9 0 .7 % 8 9 .2 % 105% 102% 104% 94.0% 97.2% 101% 91.2% 87.4% 85.9% 100% 99.5% 105% 94.4% 91.1% 84.9% 80.2% 79.0% 76.8% 116% 113% 108% 9 7 .7 % 110% 101% 8 4 .2 % 8 2 .8 % 8 5 .0 % 9 8 .1 % 9 8 .1 % 103% 104% 9 9 .3 % 8 7 .2 % 108% 9 5 .8 % 9 0 .6 % 109% 103% 101% 9 1 .3 % 101% 9 9 .0 % 8 9 .6 % 8 2 .9 % 8 4 .8 % 9 4 .7 % 9 6 .1 % 9 7 .7 % 9 3 .7 % 9 6 .5 % 8 0 .4 % 8 7 .7 % 8 2 .4 % 8 6 .3 % 8 7 .2 % 8 5 .1 % 8 8 .1 % 78.8% 8 3 .3 % 8 4 .1 % 8 4 .9 % 8 1 .5 % 8 2 .4 % 9 3 .0 % 8 7 .0 % 8 7 .7 % 8 3 .2 % 8 2 .2 % 76.5% 8 1 .7 % 8 1 .2 % 77.9% 8 5 .3 % 8 8 .8 % 8 5 .7 % 8 1 .6 % 9 3 .5 % 9 0 .2 % 8 4 .4 % 8 3 .0 % 8 1 .4 % 8 4 .3 % 8 0 .9 % 8 8 .1 % 8 6 .2 % 77.8% 74.9% 8 5 .0 % 77.8% 8 5 .6 % 9 1 .1 % 9 1 .8 % 8 8 .4 % 8 0 .5 % 101% 9 7 .5 % 9 0 .3 % 9 2 .5 % 8 6 .8 % 9 4 .0 % 8 5 .3 % 8 8 .8 % 8 8 .0 % 8 4 .4 % 79.1% 8 8 .9 % 8 5 .7 % 9 1 .5 % 30 Table 11. LCS Results A nalytical B atch ID LCS ID LCS L o w -1 ra O m LCS L o w -2 o m CL) C LCS L o w -3 a cra LCS M id - 1 <" X X CL LL LCS M id - 2 CL < C LL LCS M id - 3 CL ra 00 rsi LCS H ig h -1 o m LCS H ig h -2 LCS H ig h -3 LCS L o w -1 ra o LCS L o w -2 rsi o m a LCS L o w -3 cra < " LCS M id - 1 X X CL LL CL LCS M id - 2 <C LL CL LCS M id - 3 ra 00 rsi LCS H ig h -1 o m bo LCS H ig h -2 LCS H ig h -3 PFBA PFHx A PFHpA PFOA 96.7% 8 7 .9 % 82.1% 104% 9 5 .1 % 89.6% 97.7% 9 0 .0 % 84.9% 98.7% 9 3 .7 % 82.0% NA NA 84.8% NA NA 74.0% NA NA 78.7% NA NA 84.5% NA NA 83.9% 101% 8 0 .2 % 88.8% 102% 9 2 .6 % 87.7% 96.2% 8 3 .2 % 82.9% 96.6% 8 3 .8 % 79.5% 120% 9 8 .4 % 103% 100% 8 6 .7 % 84.3% 110% 8 7 .9 % NA 111% 9 3 .0 % NA 105% 8 5 .6 % NA 90.3% 97.6% 92.4% 87.8% 93.1% 80.9% 83.2% 90.7% 89.7% 92.0% 91.3% 91.9% 83.4% 102% 87.3% NA NA NA PFBS 87.2% 95.2% 87.0% 85.9% 88.6% 78.9% 79.6% 87.3% 85.1% 89.6% 86.5% 81.0% 76.6% 92.1% 80.0% NA NA NA ISO12-01-01-04 Tennessee River Fish Study Fall 2012 A n a ly te R ecovery (% ) PFHS T o talP FO S [b] L-PFO S [b] FBSA FOSA M eFO SA A E tF O S A A 13C 4-P F O A 13C 4 -P F O S 8 5 .2 % 9 1 .3 % 8 8 .3 % 8 2 .2 % 8 8 .3 % 78.4% 8 0 .4 % 8 4 .7 % 8 5 .7 % 8 9 .3 % 8 5 .4 % 8 5 .1 % 78.5% 9 4 .4 % 8 2 .2 % NA NA NA 86.3% 91.3% 90.6% 85.8% 93.5% 80.7% 94.9% 101% 101% 97.7% 91.2% 94.1% 98.1% 116% 96.8% 130% 148% 105% 8 5 .4 % 9 1 .3 % 8 9 .4 % 8 6 .1 % 9 3 .3 % 8 0 .1 % 8 3 .8 % 8 9 .3 % 9 0 .0 % 8 9 .6 % 8 2 .9 % 8 5 .7 % 8 9 .2 % 106% 8 7 .9 % 75.6% 79.0% 67.4% 93.3% 96.6% 96.8% 87.8% 93.2% 80.4% 76.8% 81.2% 79.5% 125% 116% 106% 86.8% 100% 93.8% NA NA NA 114% 117% 9 7 .3 % 9 9 .7 % 102% 9 5 .2 % 8 8 .4 % 105% 8 6 .4 % 8 8 .8 % 8 4 .1 % 8 4 .8 % 8 1 .3 % 9 6 .7 % 8 3 .3 % 75.1% 77.7% 71.0% 9 3 .1 % 9 6 .0 % 100% 8 9 .6 % 9 6 .9 % 8 3 .5 % 8 4 .8 % 8 5 .2 % 8 8 .4 % 8 7 .1 % 8 5 .6 % 8 7 .4 % 8 1 .2 % 9 4 .4 % 8 1 .6 % 77.9% 76.9% 74.3% 8 4 .2 % 9 3 .6 % 8 7 .7 % 8 2 .6 % 8 7 .7 % 79.2% 77.8% 8 3 .4 % 8 0 .1 % 8 1 .3 % 78.2% 78.1% 75.0% 9 2 .7 % 77.6% 76.4% 76.1% 72.3% 8 0 .4 % 8 5 .2 % 8 5 .1 % 8 8 .1 % 8 2 .8 % 74.9% 76.4% 8 4 .7 % 8 3 .7 % 8 0 .3 % 75.7% 8 1 .8 % 8 0 .5 % 8 8 .5 % 77.9% 8 3 .6 % 8 0 .6 % 74.8% 8 6 .4 % 8 6 .5 % 8 6 .3 % 9 1 .0 % 8 8 .6 % 79.3% 79.6% 8 6 .3 % 8 8 .5 % 8 1 .3 % 74.2% 79.6% 8 1 .2 % 9 1 .1 % 75.6% 8 3 .1 % 8 6 .5 % 78.8% 31 Table 11. LCS Results A nalytical B atch ID LCS ID LCS L o w -1 ra O rsi LCS L o w -2 o m a LCS L o w -3 cra <" X LCS M id - 1 X CL LL CL LCS M id - 2 < C LL CL LCS M id - 3 ra o rsi LCS H ig h -1 o m E LCS H ig h -2 LCS H ig h -3 LCS L o w -1 ra o rsi LCS L o w -2 o m bo a LCS L o w -3 cra < " LCS M id - 1 X X CL LL CL LCS M id - 2 < C LL CL LCS M id - 3 ra o rsi LCS H ig h -1 o m bo LCS H ig h -2 LCS H ig h -3 PFBA PFHx A PFHpA PFOA 87.5% 8 4 .1 % 85.0% 88.1% 8 7 .2 % 84.7% 96.4% 9 3 .6 % 95.9% 89.4% 8 7 .1 % 89.8% 95.2% 9 1 .4 % 89.8% 95.2% 9 2 .4 % 94.4% 80.9% 9 3 .4 % 75.9% 81.4% NA 80.9% 88.1% NA NA 110% 9 5 .7 % 90.6% 120% 9 4 .8 % 93.0% 110% 9 3 .6 % 93.1% 113% 8 9 .4 % 94.6% 99.1% 78.8% 85.0% 110% 8 3 .4 % 91.5% 104% NA 87.2% 124% NA NA 112% NA NA 90.8% 95.1% 99.5% 98.6% 99.9% 100% 78.0% 76.8% 93.4% 93.0% 104% 102% 99.1% 83.8% 98.0% 87.1% 100% 90.3% PFBS 83.6% 91.2% 96.5% 86.2% 99.0% 88.5% NA NA NA 92.7% 100% 96.7% 90.9% 83.8% 93.0% NA NA NA ISO12-01-01-04 Tennessee River Fish Study Fall 2012 A n a ly te R ecovery (% ) PFHS T o talP FO S [b] L-PFO S [b] FBSA FOSA M eFO SA A E tF O S A A 13C 4-P F O A 13C 4 -P F O S 8 5 .9 % 8 5 .7 % 9 3 .5 % 8 6 .1 % 9 4 .5 % 9 4 .2 % 73.5% 71.3% 8 8 .5 % 9 6 .3 % 100% 9 5 .1 % 9 4 .7 % 8 1 .6 % 9 6 .6 % 8 2 .8 % 9 3 .4 % 8 4 .4 % 102% 107% 112% 99.4% 105% 105% 100% 88.7% 104% 101% 104% 98.4% 97.8% 85.9% 99.9% 107% 115% 98.3% 101% 104% 108% 9 5 .4 % 101% 100% 75.9% 70.0% 8 0 .8 % 9 6 .8 % 9 9 .5 % 9 2 .5 % 9 3 .1 % 8 1 .9 % 9 5 .2 % 77.8% 8 6 .0 % 75.6% 80.5% 8 4 .8 % 82.2% 8 1 .9 % 99.7% 9 4 .5 % 93.8% 9 0 .6 % 103% 9 3 .7 % 93.5% 9 4 .4 % 85.5% 9 2 .5 % 85.6% 8 9 .6 % 103% 9 6 .6 % 93.4% 9 0 .4 % 99.5% 94.8% 105% 9 4 .1 % 95.5% 9 4 .2 % 88.2% 8 1 .4 % 104% 9 3 .9 % 85% 109% 103% 126% 99.80% 118% 9 0 .8 % 9 5 .8 % 100% 8 2 .3 % 9 2 .3 % 8 9 .5 % 74.5% 75.9% 78.1% 9 1 .0 % 103% 9 6 .7 % 9 2 .5 % 79.0% 9 3 .5 % 86.0% 9 4 .3 % 8 1 .4 % 8 0 .4 % 79.9% 8 5 .8 % 8 1 .3 % 8 2 .6 % 8 6 .8 % 6 5 .3 % [a] 6 3 .2 % [a] 73.5% 9 0 .2 % 9 2 .9 % 9 0 .9 % 8 8 .1 % 74.9% 8 9 .1 % 71.0% 8 0 .3 % 79.4% 77.5% 8 8 .7 % 9 2 .9 % 9 0 .1 % 9 8 .3 % 8 8 .8 % 71.6% 70.5% 78.7% 78.6% 9 2 .1 % 9 0 .0 % 8 0 .7 % 77.5% 8 3 .0 % 8 1 .7 % 8 6 .8 % 8 5 .7 % 8 1 .5 % 9 4 .7 % 9 3 .1 % 8 5 .3 % 104% 9 3 .7 % 76.9% 76.1% 8 3 .8 % 8 8 .6 % 8 7 .4 % 8 7 .9 % 8 1 .5 % 8 0 .0 % 8 3 .6 % 8 6 .2 % 9 5 .1 % 8 6 .4 % 32 Table 11. LCS Results LCS ID PFBA PFHx A PFHpA PFOA PFBS ISO12-01-01-04 Tennessee River Fish Study Fall 2012 A n a ly te R ecovery (% ) PFHS T o talP FO S [b] L-PFO S [b] FBSA FOSA M eFO SA A E tF O S A A 13C 4-P F O A 13C 4 -P F O S A nalytical B atch ID L o w LCS A v e ra g e Low -LC S S td. D ev. L o w LCS RSD L o w LCS (n ) 105% 8.1% 7.7% 30 9 5 .0 % 6 .0 % 6 .3 % 30 90.6% 12% 14% 30 95.0% 11% 12% 30 94.8% 6.9% 7.3% 30 93.5% 7.1% 7.6% 30 99.5% 9.4% 9.4% 30 9 6 .2 % 7.9% 8.3% 30 102% 17% 17% 30 9 7 .0 % 9.8% 10% 30 9 8 .1 % 6 .6 % 6 .8 % 30 8 9 .9 % 7 .5 % 8 .4 % 30 M id -L C S A v e ra g e 104% 9 5 .5 % 90.9% 96.8% 92.5% 92.0% 98.1% 9 4 .9 % 95.7% 9 6 .5 % 9 4 .5 % 8 7 .5 % M id -L C S S td. D ev. M id -L C S RSD 8.1% 7.8% 8 .5 % 8 .9 % 10% 11% 8.8% 9.1% 9.5% 10% 7.9% 8.6% 11% 11% 9.5% 10% 18% 19% 7 .6 % 7 .8 % 8 .5 % 9 .0 % 8 .7 % 9 .9 % M id -L C S RSD (n ) 28 28 30 30 30 30 30 30 30 30 30 30 H igh-LC S A v e ra g e H igh-LC S S td. Dev. H ig h LCS RSD H ig h LCS (n ) 103% 13% 13% 12 9 6 .8 % 9 .1 % 9 .4 % 7 84.9% 3.5% 4.2% 18 89.0% 8.2% 9.2% 24 84.8% 7.1% 8.3% 18 81.8% 9.5% 12% 27 106% 12% 11% 27 8 3 .7 % 11.7% 14% 30 82.3% 20% 25% 27 9 3 .3 % 13% 14% 24 8 6 .5 % 8 .5 % 9 .8 % 30 8 1 .0 % 9 .5 % 12% 30 Overall Study Average 104% 95.4% 89.4% 93.9% 91.6% 89.4% 101% 91.6% 93.6% 95.8% 93.0% 86.2% Standard Deviation 9.0% 7.4% 10% 10% 8.9% 9.6% 11% 11% 20% 10% 9.2% 9.3% RSD 8.6% 7.8% 12% 11% 9.7% 11% 11% 12% 21% 11% 9.9% 11% n (Total N u m b e r o f LCS 70 65 78 84 78 87 87 Results) 90 87 84 90 90 Analytical M etho d U ncertainty (95%CI) + 18% + 15% + 21% + 20% + 18% + 19% + 22% + 23% + 40% + 20% + 18% + 19% [a ] R ecovery w as o u ts id e o f 1 00 + 30% , b u t d ata used in s u m m a ry s ta tistics. [b ] PFOS q u a n tifie d w it h L-PFOS c a lib r a t io n , LCS re c o v e r ie s d e t e r m in e d f o r b o th L-PFOS a n d to ta l-P F O S ( lin e a r + b ra n c h e d is o m e r s ) ; all QC s a n d s a m p le s q u a n tifie d f o r to ta l-P F O S N A ; N o v a lu e d e t e r m in e d f o r th e LCS e it h e r d u e t o in s t r u m e n t e r r o r o r v a lu e o u ts id e o f th e c a lib r a t io n c u rv e ra n g e f o r t h a t b a tc h 8 7 .2 % 7.4% 8.4% 30 8 8 .0 % 9.4% 11% 30 8 4 .3 % 7.8% 9.2% 30 86.5% 8.3% 9.6% 90 + 17% 8 9 .0 % 6.7% 7.5% 30 8 9 .9 % 8.8% 9.7% 30 8 8 .3 % 6.3% 7.1% 30 89.1% 7.3% 8.2% 90 + 15% 33 6.6 Laboratory Matrix Spikes (LMSs) IS 0 12-01 -01 -04 Tennessee River Fish Study Fall 2012 As additional quality control for the study, laboratory matrix spike (LMS) samples were prepared for ~10% of the study samples. Approximately every 10thfish received in the series was prepared with four replicate weighings. The first two replicates were prepared as analytical sample and sample duplicate, the third replicate was fortified with nominal 5 ng/g (ww) of each target analyte prior to extraction as a low-LMS and the fourth replicate fortified with nominal 250 ng/g (ww) of each target analyte as a high-LMS. All four replicates were then fortified with ISs, SRSs and carried through the extraction and analysis procedure. In total, 21 of the 175 fish samples received in the study (12%) had a set of LMSs prepared and analyzed. The LMS recovery results are summarized in Table 2. The PFOS fortified into LMSs was a mixture of linear and branched isomers (br-PFOS from Wellington Laboratories), and were quantified for total PFOS using L-PFOS calibration, as described earlier. For sample analytes PFBA, PFHpA, PFOA, PFBS and PFHS and most samples for EtFOSAA and MeFOSAA, the results for the nominal 5 ng/g (ww) low-LMS were appropriate for evaluating recovery of the target analytes based on the low level of endogenous target analyte concentration in the equivalently non-fortified samples (i.e. appropriate fortification is considered that between 0.5-times to 20-times the endogenous level). For many of the LMS sample results for PFOS and FBSA the high- LMS level was the more appropriate fortification level relative to the endogenous analyte level in the non-fortified samples for determining the LMS recoveries. The overall average LMS analyte recoveries for all target analytes was typically within 100 + 10%, except for FBSA which was 82.3%. The RSDs of LMS recoveries were also < 15%, except FBSA which had an RSD of 27%. LMSs for FBSA in runs m130122b, m130129b, be130121b and j130130a were outside of 100 + 30%, but within 100 + 50% and reported with expanded uncertainties of + 50% for those affected results in Table 1. The LMS recoveries were calculated using the following equation: LMSRecovery - (Determine! Concentraionof LM S- DeterminedConcentraionof FieldSample)* 100% SpikeConcentraiton 6.7 NIST SRM 1947 Results As added QC for each analytical batch, two accurately weighed (nominal 0.5 gram) aliquots of NIST standard reference material (SRM) 1947 were extracted and analyzed the same as samples. The results of the analysis of the NIST SRM 1947 showed overall average + standard deviation (n = 10 batches, 20 results) for PFOS at 5.19 + 0.58 ng/g (ww). This PFOS result was in acceptable agreement with the NIST value and accurate to 88.0% + 10% with respect to the NIST SRM 1947 reference value of 5.9 + 0.39 ng/g. 6.8 Fillet Equipment Aqueous Rinseate Results As part of the filleting process, the equipment was rinsed and at times and the rinseates collected for analysis. Ten aqueous equipment rinseate blanks were collected and shipped with the fish samples for analysis of the target analytes. Those samples were analyzed by 3M Environmental Laboratory method ETS-8-044.1 and showed rinseates were generally less than LLOQ except 1.4 ng/L PFOS measured in sample EBLK11 and PFBA measured at 5.69 and 5.09 ng/L in samples EBLK03 and EBLK005. The results for the aqueous rinseates are provided in Table 12. Overall evaluation of rinseate blank results will be reserved for Weston Solutions, Inc. in their final overall study report. 34 ISO12-01-01-04 Tennessee River Fish Study Fall 2012 Table 12. Concentration Results for Fillet-Process Equipment Rinseate Blanks SAMPLE ID LIMS ID Concentration (ng/L; parts per trillion) PFBA PFHxA PFHpA PFOA PFBS PFHS PFOS* FBSA FOSA MeFOSAA EtFOSAA EBLK01 EBLK02 EBLK03 EBLK04 EBLK005 EBLK006 EBLK07 EBLK08 EBLK09 EBLK10 EBLK11 IS O 1 2 -0 1 -0 1 -0 4 -0 1 9 IS O 1 2 -0 1 -0 1 -0 4 -0 3 6 IS O 1 2 -0 1 -0 1 -0 4 -0 5 4 IS O 1 2 -0 1 -0 1 -0 4 -0 7 1 IS O 1 2 -0 1 -0 1 -0 4 -0 8 9 IS O 1 2 -0 1 -0 1 -0 4 -1 0 7 IS O 1 2 -0 1 -0 1 -0 4 -1 2 5 IS O 1 2 -0 1 -0 1 -0 4 -1 4 3 IS O 1 2 -0 1 -0 1 -0 4 -1 6 1 IS O 1 2 -0 1 -0 1 -0 4 -1 7 9 IS O 1 2 -0 1 -0 1 -0 4 -1 8 6 < 5 .0 0 < 5 .0 0 5.69 < 5 .0 0 5.09 < 5 .0 0 < 5 .0 0 < 5 .0 0 < 5 .0 0 < 5 .0 0 < 5 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 4 .8 0 < 4 .8 0 < 4 .8 0 < 4 .8 0 < 4 .8 0 < 4 .8 0 < 4 .8 0 < 4 .8 0 < 4 .8 0 < 4 .8 0 < 4 .8 0 < 5 .0 0 < 5 .0 0 < 5 .0 0 < 5 .0 0 < 5 .0 0 < 5 .0 0 < 5 .0 0 < 5 .0 0 < 5 .0 0 < 5 .0 0 < 5 .0 0 < 5 .0 0 < 5 .0 0 < 5 .0 0 < 5 .0 0 < 5 .0 0 < 5 .0 0 < 5 .0 0 < 5 .0 0 < 5 .0 0 < 5 .0 0 <5.00 < 0 .9 2 7 < 0 .9 2 7 < 0 .9 2 7 < 0 .9 2 7 < 0 .9 2 7 < 0 .9 2 7 < 0 .9 2 7 < 0 .9 2 7 < 0 .9 2 7 < 0 .9 2 7 1.40 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 2 .0 0 < 9 .9 8 < 9 .9 8 < 9 .9 8 < 9 .9 8 < 9 .9 8 < 9 .9 8 < 9 .9 8 < 9 .9 8 < 9 .9 8 < 9 .9 8 < 9 .9 8 < 1 5 .0 < 1 5 .0 < 1 5 .0 < 1 5 .0 < 1 5 .0 < 1 5 .0 < 1 5 .0 < 1 5 .0 < 1 5 .0 < 1 5 .0 < 1 5 .0 *R e s u lts o f LCSs p re p a re d in w a te r d u rin g th e a n a lysis in d ic a te d an a v e ra g e + RSD a c c u ra c y f o r PFOS o f 9 6 .9 % + 3 .5 % a nd w ith an a n a ly tic a l m e th o d u n c e rta in ty (9 5 % CI) o f + 17% ; a v e ra g e + RSD a c c u ra c y fo r PFBA w a s 1 00 % + 0 .5 8 % w ith an a n a ly tic a l m e th o d u n c e rta in ty (9 5% CI) o f + 16% . 35 IS 0 12-01 -01 -04 Tennessee River Fish Study Fall 2012 7 Conclusion The determined concentrations of perfluorinated chemicals (PFCs) measured in fish collected from the Tennessee River in the Fall of 2012 were reported. These analyses were conducted as 3M Environmental analytical project ISO12-01-01-04 and were performed in support of the Weston Solutions, Inc. study plan, titled "Revised Work Plant for Fall 2012 Fish and Surface Water Sampling in the Tennessee River", and dated October 2012. During the Fall of 2012, Weston Solutions, Inc. personnel caught five largemouth bass and five catfish from each of fifteen pre-defined fish sampling reaches located within the confines of the Wilson and Wheeler Reservoirs of the Tennessee River, located near Decatur, AL. The fish were filleted (skin removed) in the field. A total of 150 left side fillets and 15 randomly selected right side fillets (as field duplicates) were removed and then shipped to TestAmerica (Burlington, VT) for homogenization of the fillets. Frozen fillet homogenates were then shipped on dry ice to the 3M Environmental Laboratory for extraction and quantitative analysis of PFCs. An additional six process controls for evaluating the filleting process, three process controls for evaluating the homogenization process, and one NIST fish tissue standard reference material (SRM 1947) were also received. A total of 175 fish tissue samples were received and quantitatively analyzed. The PFCs analyzed for were perfluorobutanoate (PFBA), perfluorohexanoate (PFHxA), perfluoroheptanoate (PFHpA), perfluorooctanoate (PFOA), perfluorobutane sulfonate (PFBS), perfluorohexanesulfonate (PFHS), perfluorooctanesulfonate (PFOS) , perfluorobutanesulfonamide (FBSA), perfluorooctanesulfonamide (FOSA),2-[Nmethyl-FOSA]acetic acid (MeFOSAA) and 2-[N-ethyl-FOSA]acetic acid (EtFOSAA). Each sample was extracted in duplicate for determination of the average PFC concentrations, and approximately every10th fish sample (21 in total; 12% of total fish) had two laboratory matrix spike (LMS) samples prepared from a third and fourth 0.5 gram aliquot and fortified with known quantities of target analytes. The LMS recovery results are shown in Table 2 and average +standard deviation recovery results were within 100 +10% for all analytes, with RSDs less than 15%, except FBSA which had a recovery at 82.3% + 22% (RSD = 27%). Based on the LMS recovery results, one PFBS result for sample MSF011 (ISO12-01-01-04-011) and PFOA and PFBS results for fish MSF043 (ISO12-01-01-04-043) were not reported (NR) due to LMS recoveries outside of 100 + 50%. Laboratory control spikes (LCSs) were prepared in control bluegill fillet tissue at 3 levels, each level in triplicate, and analyzed with each analytical batch. Average LCS recoveries were within 100 + 14% at all LCS levels, and overall, for each analyte. The LCS statistical results (for all LCS levels) were used to determine the analytical method uncertainty (95% CI) for the dataset, and were + 18% for PFBA, + 15% for PFHxA, + 21 % for PFHpA, + 20% for PFOA, + 18% for PFBS, + 19% for PFHS, + 22% for total-PFOS (linear + branched isomers), + 40% for FBSA, + 20% for FOSA, + 18% for MeFOSAA and + 19% for EtFOSAA. The NIST standard reference material SRM 1947 (Lake Michigan fish tissue) was analyzed in duplicate as part of each analytical batch. The overall average PFOS value + standard deviation was determined at 5.19 + 0.58 ng/g (ww) over ten analytical batches (n = 20 results). This result was accurate to 88.0% and in acceptable agreement with the SRM 1947 PFOS reference value of 5.9 + 0.39 ng/g (ww). Two stable-isotope labeled surrogate recovery standards (SRSs) C4-PFOA and C4-PFOS were added to each sample aliquot prior to extraction. Average SRS recoveries for all the fish tissue samples analyzed (n = 175) were 85.3% for 13C4-PFOA and 88.1% for 13C4-PFOS. The relative standard deviation (RSD) for the average SRS recoveries showed excellent method precision with RSDs of 5.2% for 13C4-PFOA and 3.8% for 13C4-PFOS. Overall, the sample results in Table 1 showed that PFOS and FBSA were detected with highest frequency in Tennessee River fish samples and with the highest concentrations measured. PFOS concentrations ranged from <0.366 ng/g (ww) to 599 ng/g (ww). FBSA 36 IS012-01 -01 -04 Tennessee River Fish Study Fall 2012 ranged from <0.250 ng/g (ww) to 567 ng/g (ww). Similarly, FOSA, MeFOSAA and EtFOSAA were detected at relatively high frequencies, but generally were much lower in concentration than PFOS and FBSA. The remaining analytes PFBA, PFHxA, PFHpA, PFOA, PFBS and PFHS were detected much less frequently and at relatively low concentrations. Data/Sample Retention All fish samples and associated project data (hardcopy and electronic) for LIMS project IS01201-01-04 will be archived according to 3M Environmental Laboratory standard operating procedures. 9 List of Attachments Attachment A: Preparation Forms, Raw Data and Chromatograms (Available Upon Request) 10 Signatures Report Approval: Cleston C. Lange, Ph.D., Principal Analytical Investigator Date W illiam K. Reagen, Ph.D., 3M Environmental Laboratory Management Date Date 37 ISO12-01-01-04 Tennessee River Fish Study Fall 2012 Attachment A PFC Results from Analysis of Bass & Catfish Control Materials Used for Fillet-Process Controls Table A1. Concentration Results for Bass & Catfish Fillet Control Materials (Right Side Fillets) SAMPLE ID Average Concentration (ng/g; w et wt.) PFBA PFHxA PFHpA PFOA PFBS PFHS PFOS* FBSA FOSA MeFOSAA EtFOSAA T N 1 2 -0 3 6 4 -1 /1 3 -L M B a ss-R ig h t S ide F ille t TN 1 2 -0 3 6 4 -1 /1 3 -L M B a ss-R ig h t S ide F ille t TN 1 2 -0 3 6 4 -1 /1 3 -L M B a ss-R ig h t S ide F ille t TN 1 2 -0 3 6 5 -1 /1 3 -C a tfis h -R ig h t S ide F ille t TN 1 2 -0 3 6 5 -1 /1 3 -C a tfis h -R ig h t S ide F ille t TN 1 2 -0 3 6 5 -1 /1 3 -C a tfis h -R ig h t S ide F ille t < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 1 .0 0 < 1 .0 0 < 1 .0 0 < 1 .0 0 < 1 .0 0 < 1 .0 0 < 0.0500 < 0.0500 < 0.0500 < 0.0500 < 0.0500 < 0.0500 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 4 .2 7 4 .1 2 3 .8 5 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .1 0 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .2 5 0 < 0 .3 5 0 < 0 .3 5 0 < 0 .3 5 0 < 0 .3 5 0 < 0 .3 5 0 < 0 .3 5 0 [a ] LMSs fo rtifie d w ith n o m in a l 1 0 n g /g o f each ta rg e t a n a ly te s w e re p re p a re d w ith th e s a m p le s a n d re c o v e rie s w e re all w ith in 1 0 0 + 20% LLOQs sh o w n are n o m in a l fo r a n o m in a l 0 .5 g tissu e sa m p le e x tra c te d and analyzed R esults are sh o w n fo r rig h t side fille ts o n ly ; re m o v e d a t th e la b o ra to ry and k e p t a t th e la b o ra to ry fo r analysis. In ta c t le ft side fille ts a tta c h e d to fish b od ie s s e n t to th e fie ld fo r fille t-p ro c e s s c o n tro ls . 38 Attachment B Preparation Forms, Raw Data and Chromatograms ISO12-01-01-04 Tennessee River Fish Study Fall 2012 (Available Upon Request) 39