Document e12gEM2Vb8Kz3LBVdqQd6Vg1M

Jean B. Sweeney Staff Vqe President 3M Environmental, Health and Safety Operations (11 * CERTIFIED MAIL April 9, 2009 Document Processing Center EPA East - Room 6428 Attn: Section 8(e) Office of Pollution Prevention and Toxics, U.S. EPA 1200 Pennsylvania Avenue NW Washington, DC 20460-0001 p. 1 900 Bush Avenue, Building 42-2E-26 PO Box 33331 St. Paul, MN 55133-3331 651 7785488 G C;-- ~k q,, c-~,-4~ A1--;~ab,^ - NO CBI -1 3 1[:I ~ I[' 111~11 I~I I~ ~Il~ Re: TSCA 8(e) Substantial Risk Notice : Supplemental to Docket No. 8EHQ-0598-373 ; Sulfonate-based and Carboxylate-based Fluorochemicals To whom it may concern: 3M is submitting this notice to supplement its previous submissions on sulfonate and carboxylate-based fluorochemicals . 3M recently conducted fluorochemical (FC) analysis of 33 fish fillet tissues of walleye, northern pike, sauger, bluegill, smallmouth bass, and black crappie as split samples collected by the Minnesota Pollution Control Agency (MPCA) from the Mississippi River. Levels of PFOS quantified in all tissues were comparable to those reported by the MPCA (March 2009, http://www .pca.state .mn.us/publications/c-pfcl-01 .pdt) and ranged from 28.5 to 382 ng/g. In addition, levels ofperfluorodecanoic acid (PFDA), perfluoroundecanoic acid (PFUnA), and perflurododecanoic acid (PFDoA) were quantified in most of the samples . PFDA levels ranged from 0.581-10 .2 ng/g, PFUnA levels ranged from 0 .288-5 .81 ng/g, and PFDoA levels ranged from 0.271-3.38 ng/g. The PFOS data have been finalized and are provided in the enclosed report . Data for the other analytes are currently being finalized, and will be forwarded to EPA upon completion of the' ' reports . While 3M does not believe that any of these data taken alone or cumulatively meet the "substantial risk" reporting threshold, we nevertheless recognize the ongoing work by U.S. EPA to assess FC exposure pathways. Therefore, we are placing these results in the 8(e) docket as a supplement to previous submissions . If you have any questions or would like any additional information, please contact Deanna Luebker at (651) 737-1374 or diluebker s mmm .com. Sincerely, Jean B. Sweeney Staff Vice President, 3M Environmental, Health and Safety Operations Enclosure 3M ENVIRONMENTAL LABORATORY REPORT NO. E09-0053 Final Analytical Report Perfluorochemical Analysis of Fish Fillet Homogenates collected from the Mississippi River by the Minnesota Pollution Control Agency Phase I: PFOS Laboratory Request Number: E09-0053 Method Requirement: ETS 8-45.0 Testing Laboratory 3M EHS Operations 3M Environmental Laboratory 3M Center Building 260-5N-17 Maplewood, MN 55144 Requester Gary Hohenstein 3M EHS Operations 3M Center Building 224-5W-03 Maplewood, MN 55144 ACCREDtTED The testing reported herein meet the requirements of ISOIIEC 17025-2005 "General Requirements for the Competence of Testing and Calibration Laboratories", in accordance with the A2LA Certificate #2052-01 . Testingthat complies with this International Standard also operate in accordance with ISO 9001 :2000. Certificate #2052-01 3M EM/IRONMENTAL LABORATORY PAGE 1 OF 31 3M ENVIRONMENTAL LABORATORY REPORT NO . E09-0053 3M Environmental Laboratory 3M Environmental Laboratory Manager: William K. Reagen, Ph .D . 3M Principal Analytical Investigator and Report Author. Michelle D. Malinsky, Ph .D . Analytical Report E09-0053 Phase I Perfluorochemical Analysis of Fish Fillet Homogenates collected from the Mississippi River by the Minnesota Pollution Control Agency Phase I : PFOS Aprii 1, 2009 The Minnesota Pollution Control Agency (MPCA) submitted thirty-three fish fillet homogenates to the 3M Environmental Laboratory for perfluorochemicat analysis under project number E09-0053 . The samples were extracted and analyzed using method ETS 8-45 .0 "Determination of Fluorochemicals via Protein Precipitation of Fish Tissues (Fillet or Whole Body) and Analysis by High Performance Liquid Chromatography with Tandem Mass Spectrometry". The target analytes for this study included perfluoroalkane carboxylic acids ranging from C4 to C12, perfluoroalkane sulfonates (C4, C6, and C8), and perfluorooctanesulfonamide . This phase I report summarizes the results for perfluorooctane sulfonate (PFOS) only . The results for the other target analytes will be issued in a phase II report. The 3M Environmental Laboratory maintains A2LA accreditation to ISOIIEC 17025 for the specific tests/calibrations listed in A2LA Certificate #2052-01 and meets the relevant quality systems requirements of ISO 9001 :1994. ETS 8-45 .0 falls under the laboratory's current scope of accreditation. The data and final report were audited for compliance to laboratory's ISOIIEC17025 quality system . A key for the sample descriptions is provided in Section 2.2 . Table 1 below summarizes the PFOS results. All results for quality control samples prepared and analyzed with the samples will be reported and discussed elsewhere in this report . ACCREDITED The testing reported herein meet the requirements of ISOIIEC 17025-2005 "General Requirements for the Competence of Testing and Calibration Laboratories", in accordance with the A2LA Certificate #2052-01 . Testing that complies with this International Standard also operate in accordance wfth ISO 9001 :2000. Certificate #2052-01 3M ENVIRONMENTAL LABORATORY PAGE 2 OF 31 Table 1. Sample Results Summary: PFOS 3M LJMS ID Sample Descrf tion E09-0053-001 MissR_poo12 WE_1 --E09-0053-00.1 . .DuP._._-. __-... Average %RPD E09-0053-002 MissR-poot2-WE_2 _ E09-0053-002-Dup---- --- .----- _ Average-_ . . . .----- %RPD E09-0053-003 MissR_poo12 WE_3 E09-0053-003 Average %RPD __ E09-0053-004 MissR_poo12 WE,_4 --E09-0053-004-Du.P.__ ._._.__....__ ......__...__. ... Average %RPD E09-0053-005 MissR_pool2_SAG 1 _..E09-0053-005.Dup . .--- .. ._ _. ...... . . . ...... .. .. . . . . _ . Average %RPD E09-0053-006 MissR_poo12_SAG2 E09-0053-006 Dup-- -------- Average %RPD E09-0053-007 MissRj)ool2 SMB_1 ---E09-0053-007.. .DuP ._..._. . -__ . . . ._ Average %RPD E09-0053-008 MissRjooo12 SMB 2 E09-0053-008 Dup-----. ...--- .. .. g Avera e %RPD E09-0053-009 E09-0053-009 Dup MissR_pool2 SMB 3 _ ., .. --------- Average %RPD E09-0053-010 MissR_poo12_SMB 4 E0-9. ._-..0._0._.5_.3,- .-. ..0.-1--0.... D. ._u. p g Avera e .RPD PFOS n)Conc. 28 .7 28 .4 28.5 1.0 45.9 43.4_ 44.7 5.8 84 .3 81 .9 83.1 29 51 .0 4a .C 47.0 17 41 .1 38_6 39.8 6.3 43 .7 43 .4 43.5 0.68 57 .5 50 .4 54.0 13 0379 ---------9-)-3_8-4__._ . ..-0382 1 .4 111 123 117 10 131 122 127 7.1 3M ENVIRONMENTAL LABORATORY REPORT NO . E09-0053 3M ENVIRONMENTAL LABORATORY PAGE 3 OF 31 Table 1 Continued. 3M UMS 1D Sam le Descri 6on PFOS ~'~Conc. n E09-0053-011 MissR_pool2 SMB 5 141 E09-0053-011 Dup-_-_,- -,-----,. 150 Average 146 %RPD 6.2 E09-0053-012 MissR_pooi2 BKS_1 55 .5 E09-0053-012,Dup .__..__ ._... . .. ._ ..._...__. .___...--_--__ ._...__- . .--.._--...--.--_-~.:..5 ._. . . . . ._. .. Average 55.0 %RPD 1.8 E09-0053-013 MissR_poo[2 BGS_1 155 _E09-0053-013..Dup . . . . . .._ .. .__ ._._ ..._ . .... .. ._ . _-..____ ..... . 168 Average 161 %RPD 7.8 E09-0053-014 MissRj>ool2 BGS 2 110 E09-0053-014 D.u. F...._ . . . . __._ ._._ .____ 1 Average 105 %RPD 9.4 E09-0053-015 MissRjoo12 BGS 3 188 _ E09-0053-015Du.P ....... . . . ..----._._ ._._.... . . . .._....--------- .._. . . .__, . .._ .. . . . . ... ._.. . . . .-..---------167...... ...... . ...... Average 177 %RPO 11 E09-0053-016 MissRj)oot2 BGS_4 _E09-0053-016_Dup._.__. .__ ..__ ...----_------_--_- ._._ . _ . Average 121331 ~'340 0336 %RPD 2.7 E09-0053-017 MissR_pool2_BGS_5 123 E09-0053-017-Dup.. . . . ......_ . ... .. . . . . .__ . ._. .. ._... .__ ..._... _.._ ._... ... . . . .__. ._... ._ . ._ . .._._ . 120 Average 122 %RPD 2.0 E09-0053-018 MissR_pool3_WE_1 97 .4 E09-0053-018 .Dup. . . . ._ _. . . ..... .. .-.. ... . . . ._. 95.5 Average 96.4 %RPD 2.0 E09-0053-019 MissR_pool3 WE_2 E09-0053-0_19.Dup . . . . . . . . ._ . ._.. ... . .. . . . _------------- -Average 78 .3 ------ 78 .6 78.5 %RPD 0.37 E09-0053-020 MissRjool3 WE_3 97 .4 E09-0053-020. .DuP. .. . . .. _. . ...__ ._.. ... . . ._ _. _. 82 .3 Average ~89.9 %RPD 17 E09-0053-021 MissR_pool3 WE 4 E09-0053-02.1-Dup--.. . . ..... . . . .... ... .. ...... . . . ...._. ._. .... .. . ... ... . . . . . . .... .. Average 49.4 ---5~ 3---- . 50.8 %RPD 5.7 3M ENVIRONMENTAL LABORATORY 3M ENVIRONMENTAL LABORATORY REPORT NO . E09-0053 PAGE 4 OF 31 Table 1 Continued. 3M UMS ID Sample Description PFOS ("conc. (n& E09-0053-022 MissR-pool3 WE5 64 .2 _.E09-0053-022 Dup.----- --------._...._...--_---_._..__ 68 .1 Average 66.2 %.RPD 5.8 E09-0053-023 MissR-pooi3_NOP_1 49 .8 E09-0053-023 Dup 5t2 Average 50.5 YoRPD Y.8 E09-0053-024 MissR_pool3 SMB 1 29 .5 _E09-0053-024 29 .9 Average 29.7 .RPD 1 .1 E09-0053-025 MissR_pool3_SMB 2 103 _E09-0053-025-D4p--- ._...---._ . .......---.._.... . ._..------- 108 Average 105 %RPD 4 .9 E09-0053-026 MissR_poor3-SMB 3 48 .8 _.E...0--9---0--0-5-3----0--26...__D. . ....u._ .P.__ .. .----.._.....---....----__..... . ._.__ .._ _ ------ -- ------~.3..-----',i Average 48.5 ! %RPD 1 .0 ' E09-0053-027 MissR_pool3 SMB 4 63 .0 ' E09-0053-027 Dup--..___ __._ . . . .__._ . . ._._ ... ._-...._.._-__.._..._ . ._.._. ...._ ._ ..._. ._... ._. _ 66 .9 . . ' ---- Average 64.9 ' %RPD 6.0 E09-0053-028 MissR_pool3 SMB 5 55.0 _ E09-0053_028-DuP . ._..._ . ...._. . .___._.. .-__--._- .____-__-_-.___..__.. . . . .-_. ....... ._ . ..._._53 .3------i Average 54 .2 %RPD 3.1 ' E09-0053-029 MissR_pool3 BGS 1 76 .7 ...E09-0053-029-Du.P__..._ ._._..---..-.- .--.. . ... . _. Average _. ._ . .--------------7-2_..8_... . .... . . ...... . I 74.7 %RPD 5.3 E09-0053-030 MissRj)ool3 BGS 2 160 E09-0053-030_DuP...._. -- ...... .. .... . . .._ .._ Average _ .---1-65 163 I %RPD 2.6 E09-0053-031 _E09-0053--0--3--1---- -D---u- p MissR_pool3 BGS 3 Average 138 .-----------1-2--8-._ ._.. . .___._.I 133 %RPD 7.6 E09-0053-032 MissR-pool3 BGS 4 223 Average 221 %RPD 1 .7 3M ENMRONMENTAL LABORATORY 3M ENVIRONMENTAL LABORATORY REPORT NO. E09-0053 PAGE 5 OF 31 3M ENVIRONMENTAL LABORATORY REPORT NO. E09-0053 Table 1 Continued. PFOS I~ UMS ID E09-0053-033 E09-0053-033 Dup. Sample Description MissR_poo13 BGS 5 ""Conc. (MW 126 133 Average 129 %RPD 5.7 I (1) Unless otherwise specified, the recovery of associated laboratory matrix spike (LMS) was within i00t30% which meets ETS 8-45.0 method acceptance critena. Sample results are considered accurate to within the overall analytical method uncertainty 100t15% (PFOS) . See Section 4.9 Deterrnination ofAnalytical Method Uncertaintyfor more information . Sample results are reported to three significant figures and %RPD to two significant figures. Additional significant figures were used to calculate the average, %RPD, and %recovery. Values in the raw data may vary slightly due to rounding . (2) Sample results were reported from a 1 :10 post-extraction dilution . 2.1 Target Analytes . Table 2 below provides pertinent information regarding the target analytes, internal standards, and surrogate compounds investigated in this study. Table 2 . Target Analyte Summary. Compound Name Synonym or Acronym Analytical Purpose i Formula ~Reference Standard Source Perfluorobutanoic acid Perfluoropentanoic acid Perfluorohexanoic acid Perfluoroheptanoic acid Pertluoroodanoic acid Perfluorononanoic acid i Perfluorodecanoic acid Perfluoroundecanoic acid Perfluorododecanoic acid Perfluorobutane suffonate Perfluorohexane sulfonate Perituorooctane sulFonate PFBA (C4 Acid) Target T C,F7COOH ~ PFPeA (C5 Acid) Target C,FBCOOH PFHxA (C6 Acid) Target CsFCOOH PFHpA (C7 Acid) Target ~FCOOH PFOA (C8 Acid) Target C7F, SCOOH [G~F,sCOO~[NH,7 PFNA (C9Acid) Target PFDA (C10 Acid) Target PFUnA (C11 Acid) PFDoA (C12 Acid) Target Target PFBS (C4 SulFonate ) Target -~t-- ~ PFHS (C6 Sulfonate) Targe! t I' I ' -1li - T PFOS (C8 Sulfonate) ~ Target ~- -- _ L j --- I CaFCOOH CsF,sCOOH C,oFZ,COOH C F23COOH [C4FBSO,)[K`] [C6F S0,1[Na'] [CaF,7S03-][K'] --- [GaFrtSO31[K~ Aldrich Aifa Aesar Oakwood Products Aldrich L-PFOA (linear) Wellin gt on Labs ECF PFOA (branched) 3M Production Lot 332 Oakwood Products Oakwood Products Oakwood Products Oakwood Products 3M Production Lot 2 L-PFHS (linear)Wellin on Labs L-PFOS (linear} Wellington Labs ECF PFOS (branched) 3M Production Lot 217 3M ENVIRONMENTAL LABORATORY PAGE 6 OF 31 3M ENVIRONMENTAL LABORATORY REPORT NO. E09-0053 Table 2 Continued. Compound Name Synonym or Acronym Analytical Purpose Formula Reference Standard Source Perfluorooctanesultonamide '3C4-Perfluorobutanoic acid "CrPerfluorohexanoic acid "C4-Perfluorooctanoic acid "C-Perfluorononanoic acid "CrPerfluorodecanoic acid "Cz-Perfluoroundecanoic acid "CZ-Perfluorododecanoic acid '80r-Ammonium Perfluorobutane sulfonate "02-Ammonium Perfluorohexane sulfonate "C4-Sodium Perfluorooctane sulFonate "Cz-Perfluorooctanoicacid 'OrAmmonium Perfluoroodanesulfonate FOSA (C8 SuNonamide) (1,2,3,4-"C4]PFBA [1,2-"CJPFHxA [1,2,3,4-"C4]PFOA [7,2,3,4,5-"Cs]PFNA [1,2-"CzJPFDA 11,2-"C,JPFUnA [1,2-"CzJPFDoA ['O~PFBS ['O]PFHS [1,2,3,4-"C,]PFOS [1,2-"C,]PFOA 'O PFOS Target CFSOzNH2 IS "CF,("CFZ)2"COOH IS CF,(CFZ),("CF,)"COOH IS CF,(CF2y,("CFZ),"COOH IS CFs(CF,),("CF,),"COOH IS CF,(CF2),("CFZ)"COOH IS CF,(CFs)("CF2)"COOH IS CF,(CF2W"CFZ)"COOH IS [CaF~.S'OZ01 [NH,'] IS (CFS'OZO1[NH,`) IS [CF,(CFz),("CFz},"CFZSO,'j[Na'J Surrogate CF,(CF,)s("CF,)"COOH Surrogate I (CFS'1 0~[NH,~ I 3M (L-15709) Wellington Labs Wellington Labs Wellington Labs Wellington Labs Wellington Labs Wellington Labs Wellington Labs RTllntemational Wellington Labs Wellington Labs PerkinElmer RTl International J 2.2 Sample Descriptions. The table below summarizes the abbreviations provided by the MPCA used to identify the fish species collected for this study. No information was provided regarding the location of sample collection within Pool 2 or Pool 3 of the Mississippi River. Table 3. Sample Description Key. - Species Blu iU Walle ye Black Crapp ie Sau er Smallmouth Bass Northern Pike Abbreviation BGS WE ' BKS SAG SMB NOP 2.3 Sample Collection and Receipt. The chain of custody provided by the MPCA indicated that the samples were collected on June 5 and June 9, 2008 . On January 27, 2009, the 3M Environmental Laboratory received the homogenized fillet samples in Nalgene bottles on dry ice. The samples were logged into the 3M Environmental Laboratory LIMS system under project E09-0053 and placed in frozen storage. Information provided by the MPCA indicated that the samples were homogenized with the skin on and scales removed; however, no specifics regarding the homogenization procedure were given. 3M ENVIRONMENTAL LABORATORY PAGE 7 OF 31 3M ENVIRONMENTAL LABORATORY REPORT NO. E09-0053 It should be noted that these fish samples have been previously handled and analyzed at another laboratory under contract by the MPCA . It is unknown how these samples have been handled during analysis and storage. It is possible that some, or all, of these samples may possibly have been compromised during the previous handling, analysis and storage. No assumptions are being made to the integrity of these samples. 3M Environmental Laboratory personnel removed the homogenized fish tissue from the sample container by cutting the bottle away from the frozen sample mass using a clean utility knife. The frozen sample mass was then placed in a plastic bag and retumed to frozen storage before the tissue was homogenized before extraction . Based on the sample consistency in the Nalgene bottles, it appeared that the sample homogenates had thawed at some point before they arrived at the 3M Environmental Laboratory . (Homogenized tissue had refrozen into a solid mass that could not be broken up while frozen in the container.) ~thods - Analytica'I~;arial Prepara~ltory 3 .1 Tissue Preparation Each submitted sample and bluegill fillet control tissue (TN08-0220-1/1) purchased from Osage Catfisheries, Inc . (Osage Beach, MO) were homogenized frozen using dry ice. Depending on the sample size, frozen fish tissue was added to a pre-chilled stainless steel bowl of either a Robot Coupe RS12Y-1 or a RS123B vertical batch processor and homogenized with dry ice until a fine powder consistency was reached. After homogenization, the ground fish tissue was transferred to a polyethylene bag which was placed in the freezer at -20C unsealed ovemight to allow the residual carbon dioxide to sublime. After sublimation, the bag was sealed and homogenates were kept frozen . 3.2 Sample Extraction Polyethylene centrifuge tubes (50 mL) were chilled in a cooler with dry ice for approximately thirty minutes prior to weighing homogenate aliquots to prevent the tissue powder from thawing and congealing during the weighing process. A 0.5 g sub-sample of the fish tissue homogenate was accurately weighed in a pre-chilled 50 mL polyethylene centrifuge tube . Tissue aliquots were then spiked with internal standard (IS) and surrogates to produce an approximate concentration of 1 nglg . Samples designated as lab matrix spikes or matrix-matched calibration standards were additionally spiked with the target and surrogate analytes at appropriate levels to produce the desired tissue concentrations . Samples intended for laboratory QC (matrix blanks and laboratory control matrix spikes) were spiked with surrogates at 1 ng/g and the other target analytes at appropriate levels . After spiking, the tissue homogenates were allowed to sit for 30 minutes before 5 mL of acetonitrile was added. The samples were then thoroughly homogenized with the acetonitrile using an Omni Prep multi-place homogenizer with disposable plastic probes (15,000 rpm for 2 minutes) . The acetonitrileltissue extracts were then placed in a freezer at -20C for at least one hour . Upon removal from the freezer, sample extracts were centrifuged at a targeted temperature of -5C for 20 minutes at 3000 rpm. If any delays occurred, samples were retumed to the -20C freezer and then re-centrifuged . After centrifugation, 1 mL of clarified supematant was transferred to a 2 mL autovial spiked with 10 pL of 10% formic acid . Autovials were then capped and vortex mixed. Samples were extracted in three separate batches . For each submitted sample, a sample, sample duplicate, and laboratory matrix spike (LMS) were prepared . Two samples were re-extracted in a fourth batch as the associated LMSs prepared the first time were not high enough for resultant endogenous concentration. Each preparation batch consisted of the following calibration and quality control elements : 3M ENVIRONMENTAL LABORATORY PAGE 8 OF 31 3M ENVIRONMENTAL LABORATORY REPORT NO . E09-0053 a nine point extracted matrix calibration curve using the purchased bluegill fillet control tissue " triplicate lab control spikes of the bluegill fillet control tissue at three levels (0 .05 ng/g, 1 .5 ng/g, and 8 ng/g) " triplicate lab control spikes of 3M electrochemical fluorination (ECF) PFOA and PFOS at 5 ng/g " duplicate control matrix blanks with and without internal standards and surrogates " duplicate method (aqueous) blarncs with and without internal standards and surrogates The table below summarizes the preparation dates and samples prepared in each batch. Table 4. Sample Preparation Summary. Batch Number Batch 1 Batch 2 Batch 3 Batch 4 on Dates 2118/2009- 2/2012009 211912009 - 2J23l2009 ?J24l2009 - 2"2512009 3/5/2009 - 3W009 Elm&! E09-0053-001 - E09-0053-011 E09-0053-012 - E09-0053-022 E09-0053-023 - E09-0053-033 E09-0053-008 & E09-0053-016 lrenreol 3.3 Analysis Analysis for the suite of PFCs listed above was performed using high performance liquid chromatography-tandem mass spectrometry (HPLC/MS/MS) . For this investigation, samples were analyzed using an Agilent 1100 series (Palo Alto, CA) HPLC system interfaced to either a PE SCIEX API 4000 triple quadrupole or SCIEX API 4000 Q-Trap mass spectrometer (Foster City, CA). Both instruments were equipped with a SCIEX Turbo V ion-spray interface operating in the negative ion MS/MS mode using multiple reaction monitoring (MRM). AnalystT"' 1 .4 .2 software was used for all data collection and reduction. Table 5 lists the MRM transitions monitored for each analyte. A Thermo Scientific PRISM RP guard column (2 .1 mm x 50 mm ; 5p particle size) was placed in-line after the purge valve and before the sample injection port to trap any PFC contaminants coming from the HPLC instrument and/or the mobile phases . This sufficiently separated the elution of the "system" PFC peaks from those present in the sample extracts . 3M ENVIRONMENTAL LABORATORY PAGE 9 OF 31 3M ENVIRONMENTAL LABORA TORY REPORT NO. E09-0053 Table 5, MRM Transition Summary. Compound Anal e Descri tion ~^ WMRM I Transiiion s Dwell Time ms PFBA C4Acrd.1--------------------- - ------ - PFPeASC5Acid)----- --------------- - ------- - ------Target----------------~ --- -21-3->-1-6-9----------1-0-0------ - --------------Target ------------ - . . . . .2-6--3>--21--9------- -1-0--0------ 313>269 --- 100 - PFHxA(C6Acid)- ------------- - --------------Target--------------- - 13>119----------1-~----- 363>319 50 - PFHpA -- SC-7-A-c-id)- -------- - ----- Tarqet - ----------------- " ----------------- ---3-6-3->-1--6-9-----------------5-0-- - -- -4?3>369 50 413>219 50 - PFOASCSAcid) - -- ------ -------- -------- ---------------Ta rge---------------- -- :- 113- >16-9-- ---- ---- -----FO - - - - 463>419 50 463>219 50 - PFNAiC9Acid)-- . ---- ---------- ---------------Target---------------- - -- ---4-6-3>-1-6-9-----------5-0------ 513>469 50 513>269 50 P-FDA- (C10Acid)-------- --- - --- I 1 - --------------Target--------------- ~ ---5-1- 3>--21~-9---5611 >5 9 ------ 50 -50 .- .- 563>269 50 - PFIIM[C1-~--A-c--id--)------------ ------------ ---Ta--rget- - --- - - . .- . . .563>219---------- 50-- .. --- - -~ 613>569 50 -- -- 613>319 5 ,IO - PFDa4[C12-Add ------ i --------------- T--arge-t---------- ------ - ---61-3->--1-6-9_--_-_--5- 9---I --- i 299>80 50 - PFBS_(C4Suffonate).-- .~ ------------ ---Target-------- ------ - --- 299>99 --------- 50 ------ ---------- I 399>80 -50 - - PFHSfC6-S-u-I-f-o-n-a-t-e- ---- --------- .- .. - . . ..---_---Target--------------- - -----4--9-99->>-89-09----- . _ ---55-00------- ~ 499>99 50 -P---F-O-S--(CS.Suffonate) ------- .-- -- --- - -----------Target--------------- -- 499>130--------- 50 50 --- FOSA C8 Suffona_ mide Ta rg et 498>78 - IS for PFBA and PFPeA 217>1,. 2----------------1-0-0- ! [-1-,-2-,-3-,4- =--'-C-,-]-P-F--B-A---- - . -------- -------------I-S-f-o-r-P--F-H-x-A-------- - --- - - ---3-1--5->-2-7-0- - --- - ---- --' 100 [1,2 "CJPFHxA ------ - - - - - - - - --- ---- --- . .------------- --------- ---------- - ---- -------------------- ----- and PFOA 41 i>372 50 [1,2 ,3,4-"C,]PFOA I - ----- ----- ----- ---- - -- -------- IS for PFHpA - - -------- -- --- ---- - - [1,2,3,4,5-Cs]PFNA ---------- _ ~ ------------- tSforFFNA ------------------------ ---------------- i - ---468>423---------------------- -- ------50- - - - - - ---------------------- IS for PFDA 515>470 50----- i --[-1,-2---"-CJ--P--F--D--A--- --- -- -1-- - -------------------------- - --- - ------- ------ - ---- -- I [1,2 "Cz[PFUnA IS for PFUnA 565>520 50 ---------------- ------------ - - - - - - -"-C-2- - - - - - - - - - - - - -------- -----------------------------------IS for PFDoA 61 5>570 50 [1-,2---------------]PFD-oA- ---------- - - -- ----------------------- - --- - - - ------- -[--"-O-z]--P-F-B--S--- IS for PFBS -------- - --------------- -------------- ---- ~ - 303>84 ---------- - 50 ------------ - -- [ Oz]PFHS -------------------- - - ---------- - ------------ - 1-5-f-o-r-P--F-H-S------------ - ~ - 403>84 50 ---------- .--- - - --------- - [1,2,3,4- c,]P~OS IS for PFOS, FOSA ~ 503>80 50 [1,2 .13C2]Pf=OA ['OzJPFOS Surrogate (Acids ). i 415>370 - I Surrogate (Sulfonates FOSP.) ! 503>84 50 I 50 3M CNti~IROPdMENTAL I.ABORATORY PAGE 10 OF 31 p. 12 3M ENVIRONMENTAL LABORATORY REPORT NO . E09-0053 (1) Individual transitions were summed foreach analyte to produce a "total ion chromatogram" (TIC). TheTICs were used for quantitation . Analysis of the small acids (C4 through C6) was performed using a Thermo Scientific PRISM RP column (2 .1mm x 50 mm ; 5 ~L particle size) held at 30C with the mobile phase system consisting of 5 mM ammonium acetate with 0.01 % acetic acid in water (A) and methanol (B). The gradient used to elute the analytes of interest is presented in Table 6. The flow rate was held at 300 pUmin throughout the run and a 20 pL injection volume was used. The outlet of the analytical column was directed to a column switching valve where the first three minutes of the run were diverted to waste. After three minutes, the valve was switched to direct the effluent to the mass spectrometer for analysis . The large acids (C7 through C12), the sulfonates, and FOSA were analyzed using a Thermo Scientific BetasilT"" C18 analytical column (2.1mm x 100 mm ; 5p particle size) held at 30C with the mobile phase system consisting of 2 mM ammonium acetate in water (A) and acetonitrile (B). The gradient used is also provided in Table 6. The flow rate was held at 400 pUmin throughout the run and a 25 gL injection volume was used . Samples were injected onto a Waters (Milford, MA) Oasis HLB on-line extraction column (20 mm x 3.0 mm, 25 p particle size) with the outlet directed to a column switching valve where the first five minutes were diverted to waste. After five minutes, the valve was switched and the effluent was directed to the analytical column for separation of the target analytes and analysis by the mass spectrometer . Table 6. LC Gradient Parameters . Small Acids: C4-C6 La Adds: C7-C12; SuNonates, FOSA S Total T'rme min %A 0 0 :0 90 1 3 .0 90 2 3 .5 30 3 9 .0 5 .0 4 15 .0 5 .0 5 15 .1 90 6 19.0 90 Total 77rne %.B mtn %.A VO8 10 0 0 .0 10 1 3 .0 97 3 .0 97 3.0 70 2 3 .5 70 30 95 3 13 .5 40 60 95 4 15 .5 40 60 10 5 16 .0 10 6 18 .0 7 18 .3 10 90 10 90 97 3.0 8 21 .0 97 3.0 A 5 mM ammonium acetate with 0.01% acetic acid (aq) A 2 mM ammonium acetate (aq) B Methanol B Acetonitrile To manage the large number of transitions required for the large acids, sulfonates, and FOSA, a multiperiod method was constructed . The first period (time=0 minutes to 10 .6 minutes) collected the transitions for PFBS, PFHpA, PFOA, [1,2 -13CZ]PFOA, PFHS, PFNA and their respective ISs. The second period (time= 10 .5 min to 21 .0 minutes) collected the transitions for PFDA, PFUnA, PFOS, [1802JPFOS, PFDoA, and FOSA and their respective ISs. For analytes where more than one transition was acquired, a total ion chromatogram (TIC) which summed the respective transitions for that analyte was used to analyze the data . 4.1 Calibration Extracted 'matrix-matched calibration standards and QCs were prepared by spiking known amounts of target analytes, surrogates, and internal standards into individual 0.5 g aliquots of bluegill fillet control 3M ENVIRONMENTAL LABORATORY PAGE 11 OF 31 3M ENVIRONMENTAL LABORA TORY REPORT .NO. E09-0053 tissue (TN08-0202-1/1) homogenate . Each spiked fish aliquot was then extracted using the procedures outlined in Section 3.2 . A total of nine spiked standards ranging from 0 .25 ng/g to 25 ng/g (nominal) were prepared . Additionally, an unextracted acetonitrile solvent curve ranging from 0.025 ng/mL to 50 ng/mL was also prepared . All sample results for PFOS and ['g02]PFOS were quantitated against the solvent curve as sample extract concentrations for PFOS exceeded the upper calibration range of the extracted curve. To demonstrate equivalency, the extracted curve and the laboratory control QC samples were quantitated using the solvent curve. Once the concentrations were corrected for the endogenous level of PFOS in the control tissue, PFOS recoveries of the calibration curve points were within 100t14%. The recoveries of the extracted curve points quantitated against the solvent curve points are provided in the Supplemental Information Section 11 .1 . A quadratic, 1/x or 1/x2 weighted calibration curve was used to fit the data for PFOS and ['a02]PFOS . Internal standard quantitation was used . The data was not forced through zero during the fitting process. The accuracy of each curve point was verified by calcuiating the concentration using the area count ratio of the target analyte to the internal standard . Each solvent calibration standard used to generate the final calibration curve met the method calibration accuracy requirement of 100t25%, except the LOQ standard (100t30%). The coefficients of detemiination (r`) were greater than 0.990 for all analytes . As required by ETS 8-45, a minimum of six calibration points were used to generate the final calibration curve. For PFHS, PFOS, and PFOA, the reference standard used for calibration and lab control spikes was comprised predominantly of the linear isomer . It is noted that the purchased control bluegill tissue matrix differs in composition from the study bluegill samples in that the purchased bluegill fillet homogenate does not contain the skin of the fillet, only the "meat" . 'As fish matrix can vary greatly from species to species, the control matrix used to generate the extracted calibration curve and laboratory control QC samples is considered to be matrix-matched for the bluegill samples only . The laboratory control matrix should be considered as a "surrogate" control matrix for the other species investigated in this study. 4.2 Limit of Quantitation (LOQ) The LOQ as defined in ETS 8-45 is the lowest non-zero calibration standard in the curve in which the area counts of the target analyte are at least twice those of the matrix blank(s) . Since the solvent curve was used for quantitation, the aqueous method blanks were used to establish the LOQ so that the curve could be used to quantitate the endogenous levels of PFOS in the control matrix . The limit of quantitation for PFOS varied from extraction date and instrument batch. The PFOS LOQ ranged from 0.00251 ng/mL to 0.00995 nglmL which corresponds to approximately 0.025 ng/g to 0.1 ng/g in fish tissue once the conversion is made . 4.3 System Suitability Four replicate injections of the solvent calibration standard were analyzed at the beginning of the analytical sequence to demonstrate overall system suitability. Method criteria states that system suitability injections shall produce a RSD of less than 7% for the ratio of target analyte area counts to internal standard area counts and an RSD of less than 2% for the retention time . Method acceptance criteria were met for both PFOS and ['802]PFOS . 4.4 Continuing Calibration During the course of the analytical sequence, several continuing calibration verification samples (CCVs) were analyzed to confirm that the instrument response from initial calibration curve was still in control. CCV injections produced recoveries of PFOS and ['8OZ]t'FOS within 100%t25%, which met method criteria . 3M ENVIRONMENTAL LABORATORY PAGE 12 OF 31 p. 14 3M ENVIRONMENTAL LABORATORY REPORT NO. E09-0053 4.5 Blanks Five types of blanks were prepared and analyzed with the samples: matrix blanks (btuegill filet control tissue : two with IS and surrogate, two without IS and surrogate), aqueous method blanks (two with IS and surrogate, two without IS and surrogate), acidified acetonitri(e solvent blanks, acetonitrile blanks with internal standards and surrogates, and acetonitrile solvent blanks (unmodified). Each blank result was reviewed and used to evaluate method performance to determine the LOQ for each analyte. Surrogate recoveries of spiked blanks are provided in the Supplemental Information Section 11 .2 . 4.6 Lab Control Spikes (LCSs) Triplicate lab control spikes of the purchased bluegill fillet control matrix at three different levels were prepared each extraction day to determine analytical method uncertainty (See Section 4.9). For PFHS, PFOS, and PFOA, the standard reference material used for the LCS spikes was the linear isomer, which was the same used for the calibration curves . Separate 3M electrochemical fluorination (ECF) spikes of branched PFOS and PFOA were prepared to evaluate any potential bias from quantitation against a linear standard . Lab control spikes were prepared to evaluate method accuracy and precision and were used to determine analytical method uncertainty (See Section 4.9). The ['802]PFOS surrogate was spiked at a consistent 1 ng/g (0 .1 ng/mL) for all LCS samples. For PFOS, the spiked concentration was corrected for the average PFOS concentration determined for the control matrix in the same extraction batch. Lab control spike recoveries are provided in the Supplemental Information (Section 11 .3). For both PFOS and [' OZ]PFOS, all replicates (N=48) except one, produced recoveries within method acceptance criteria 100%t25% . 4.7 Sample Concentration Calculation The solvent calibration curve used for quantitation provided extract concentration in units of ng/mL. . The following equation was used to convert the extract concentration to sample tissue concentration in units of ng/g . Tissue Concentration (~g) 9 Extraction Volume = 5 mL Extracted Mass -0.5 g Extract Concentration ( 9)'ExtractionVolume(mL) Extracted Mass (g) 4.8 Lab Matrix Spikes (LMSs) For each submitted sample, a separate laboratory matrix spike was prepared by extracting a separate aliquot of the fillet tissue that was spiked with the target analytes . For each LMS, all target analytes were spiked at a nominal concentration of 2 ng/g except PFOS, which was spiked at a total concentration of approximately 150 ng/g . The recovery of the LMS was calculated using the following equation . (Extract Conc. (m9 )' ExtractionVolume(mL)1-(Average Sample Conc.( LMS %Recovery = l -- J l l g Spike Amount (ng) ' LMS Sample Mass l,g)} . ) 100% Although ETS 8-45 .0 does not provide specific criteria for the lab matrix spike concentrations, the 3M Environmental Laboratory uses a general guideline that lab matrix spike concentration should be at least half the endogenous concentration before the spike is considered "appropriate" for the given matrix . For PFOS, the LMS was 76 .0 ng (-150 ng/g tissue concentration) except for samples E090053-008 and -016 where the LMS was 250 ng (-500 ng/g tissue concentration) . The ['802]PFOS surrogate was spiked at 0.502 ng (-1 ng/g tissue concentration). 3M ENVIRONMENTAL LABORATORY PAGE 13 OF 31 3M ENVIRONMENTAL, LABORATORY' REPORT NO. EC9-Gl?5? 4.9 Analytical Uncertainty ETS 8-45.0 states that the analytical method uncertainty will be determined by statistical eva!uatior, of the individual analyte recovery in LCSs . Current project QC results, as well as historical values, are used in the evaluation until a maximum of fifty points (minimum f twenty, if possible) are included . For ETS 8-45 .0, the historical LCSs are categorized by tissue type : whole-body or fillet unless there is strong evidence suggesting that a species-specific matrix effect is present and that results from these species should be considered separately . The standard deviation of the collective recovery results (in %) is calculated . The expanded uncertainty is then calculated by multiplying the standard deviation by a factor of two, which corresponds to the 95% confidence limit . Table 7 below provides the analytical uncertainty for PFOS and the ['802]PFOS surrogate. Table 7. Analytical Method Uncertainty Average `Yo Recov Standard Deviafion _Z'Sfandard Devlatlon Ly _ Analydcal Uncentwnty PFOS 105 -_ 7 .6 15 50 (2 Rainbow Trout fillets, 48 Blu ill fillets ) t15/. - `O PFOS Su ate 108 7 .1 14 48 (Bluegill fillet only) t14% Table 8 below provides the sample and lab matrix spike results for PFOS and the ['BOI]PFOS surrogate c;uantitated using an unextracted, solvent curve. The extracted curve prepared in blueglii fillet control matrix was demonstrated to be equivalent to the solvent curve for both PFOS and the ('eO2JPFOS surrogate over the approximate range of 0.25 nglg to 25 ng/g . (See Supplemental Information Section 11 .1 for additional information .) As most of the study samples were greater than 25 ng/g for PFOS, the solvent curve, which went up to 50 ng/mL or -500 ng/g was used for quantitation . All samples produced PFOS LMS recoveries within method acceptance criteria of 190t30%. E09-0053-008 and -016 were re-prepared in a separate extraction batch with a LMS of 2.50 ng (-500 ng/g). The initial analyses for these two samples indicated that the original LMS concentration of -150 ng/g was less than half the endogenous concentration and therefore was not considered an appropriate level for assessing sample quantitation accuracy . Results for these two samples from the initial analysis were not reported . Instances of the surrogate recovery exceeding 100t30% have been flagged in Table 8; however, ETS 8-45 .0 specifies no acceptance criteria for surrogate recoveries as the intent of the surrogate is to evaluate systematic biases or interferern;es that rnay be present As the PFOS LMS recoveries were all within 10030% and the overall number of surrogate recoveries exceeding 100t30% were few, all results were considered reportable and accurate to within the stated analytical method uncertainty of 100t15%. The recoveries of the 3M ECF LCSs prepared with each extraction batch (N=12 for the study) ranged from 97 .5% to 130% with a calculated average of 110% (8.4% RSD) . The results of the ECF QC spikes demonstrated that there is no discernible bias when PFOS ECF isomers are quantitated against the predominantly linear reference standard . Therefore, the reported PFOS values are considered accurate to within the stated method uncertainty regardless of the degree of branched PFOS isomers present in the sample extracts . The PFOS sample concentrations presented in Table 1 and Table 8 were reported from the total ion chromatogram (TIC) where the instrument software summed the three individual transitions (499>80, 1M Eh1V1P.0MMENiAL LABOPATOR`l PAGE 14 OF 3 ! 3M ENVIRONMENTAL LABORATORY REPORT NO . E09-0053 499>99, 499> 130) . As the 499>80 transition has been well documented to produce PFOS matrix interferences in biological tissue studies[1], the three individual transitions were evaluated separately . to verify that the area rabos observed in the standards were conserved in the samples. For Batches 1, 2, and 3, the combined average area percentages of 499>80, 499>99, and 499>130 in the solvent calibration curve points were 76 .5% (3.4% RSD), 21 .1% (13% RSD), .and 2.36% (42%RSD), respectively. The 499>99 and 499>130 transitions are significantly weaker in intensity when compared to the 499>80 transition . The higher variabilities observed for these two transitions in the solvent curve was largely due to very weak signals in the calibration standards less than 0.5 ng/mL in concentration . Calibration standards of 0.5 ng/mL and higher produced %RSD of approximately 8% for these two transitions . Figure 1 and Figure 2 display the average area percentages observed for each transition in the study samples. Here, the plotted average and %RSD is the statistical calculations for sample, sample duplicate, and associated LMS. For viewing simplicity, the /flRSD for 499>130 was excluded from the graphs as it was often larger than the area average and produced error bars that extended into the negative y-axis region making the graph difficult to read . From the figures, it is clear that the overall transition ratios observed in the solvent standard are conserved in the extracted samples and QC . - 3M ENVIRONMENTAL LABORATORY PAGE 15 OF 31 3M ENVIRONMENTAL LABORATORY REPORT NO . E09-0053 Table 8. Sample and Lab Matrix Spike Results: - -- .- PFOS and ['e02]PFOS surrogate. PFOS '80 PFOS 3M LIMS ID Sam Descri bon Mass Conc. n L IrIConc. n % Rec Conc. n mL Conc. n. *~,-Rec E09-0053-001 MissR_pool2 VVE_1 0.4799 2.75 28 .7 NA 0.102 1 .06 102 E09-0053-001 Dup 0.4884 2.77 28 .4 NA 0.108 1 .11 108 E09-0053-001 LM S 0_5032-,- - 19 .9 198 112 -0 .126 1 .25 126 -- Average 28.5 Average %Recovery 112 %RPD 1-0 %RSD 11 E09-0053-002 MissR_pool2 WE 2 E09-0053-002 Dup _E._0.._9.-_ 0. .0_..5._3-_-. .0.-0-2---L--M.. S..._ .. _.--._-.------------- 0.5049 0.5212 -- -0.__.5. ..-0--0...8.. .. . . .--Average 4 .64 45.9 NA 4 .52 43.4 NA ---1-8--.-9 --- -----1-8-9.._._. . . .- -----9--5-.-0--44.7 0.107 1 .06 '.07 0.104 1 .00 104 0 .122 1 .22--_- 122 -.-- ----- Average %Recovery 111 %RPD 5.8 %RSD 8.7 E09-0053-003 MissR_pool2 WE 3 0.5202 8.77 84 .3 NA 0 .108 1 .04 108 E09-0053-003 Dup 0.5056 8.28 81 .9 NA 0.112 1 .11 112 _0.-_1 .5- - 1 . 1 __E..-0....9.._-_.0.._.0.-5-3---0--0-3--L-M.....S.----- _.__..._.._..-_-.___....__...._.......__..___. -_0_._5_1._5,._9..-.__.__.. ...._..-2_.4.._.....4....._..-Average %RPD E09-0053-004 E09-0053-004 Dup E_ 09-0053-004 LMS MissR_pool2 WE_4 0.4822 4.92 0.4974 4.28 0.516,3. .-._... . . .... - ---2.. .0. . .....5. . ... .. Average 236 .__- .__ ._...-_1_.0..._4. . . ......_ - . -_0_._13_ 3. . 1 .29 83.1 Average %Recovery 29 %RSD 51 .0 NA ' 0.114 1,18 43 .0 NA 0.105 1 .06 199 103 I ------- 4,'.0 I Average %Recovery M133 117 11 114 105 115 111 %RPD 17 %RSD 4.9 E09-0053-005 MissR_pool2 SAG 1 0.4871 4 .00 E09-0053-005 Dup 0 .4954 3 .82 E0_9. .-..0. ....0..5. . .-3_-0. ...0--5--.L. .. .M. ...S.. . ..._... . . ... .... . .. .... .. _... . ........ ......... . . ... ... . .. ... .0 ;4781._ . .,.._. . . .. . . . . . .. t8.4 Average 41 .1 NA 0.101 1 .04 101 38 .6 NA 0.106 1 .07 106 __1. . .9_~._.. ... . .--- . . ....- .9--6-- ..1. . . . .. _. ..-._0_..1_ .0.7. . .__ .. . .. . .. .__-1.-.._.1..2... .-----. .. . . ... . 1.--0--7-... . . .. 39.8 Average %Recovery 104 IoRPD 6.3 %RSD 3.1 E09-0053-006 MissR_pool2 SAG 2 0.4808 4 .20 43 .7 E09-0053-006 Dup 0.5129 4 .45 43 .4 E09-0053-006 LMS . , . . .... . . . . . ._ .. . ._ . ._ . . ._..- .. . . . . .. .- .. . 0.48.9-1 .. . . . ... . . . 18. 3 . . . 187 Average 43-5 %RPD 0.68 NA 0 .0999 1 .04 100 NA 0.114 1 .11 114 '-9--2-.4. .. . ..-- -0.-1- -1 - . . .. 1-13 111 Average %Recovery 108 %RSD 6.9 E09-0053-007 MissR-poo12 SMB 1 0.5287 6 .08 57 .5 NA 0.105 0 .99 105 E09-0053-007 Dup 0 .5257 5 .30 50.4 NA , 0.109 1 .04 109 E_.0. .-9-._0. ..0_-5-3..-__0..0...7---.L._ ..M_ ..S.__-__ ._....._-.- ..... .__ . ._ . . ....---._ .__. ._... ... _--.0. ..__4. 9.4_3... --- ... .,._- . 2. . . .2.-....9___ . . . --_---2__3.2.___,.._.- . ._. ._ . 1._1.,6----- ---0_.-1-16. . .-_..___- _ 1 17 116 Average 54.0 Average %Recovery 110 E09-0053-008 E09-0053-008 Dup E09-0053-008LMS MissR_pool2 SM8 2 `.'oRPD 0 .4785 0 .5163 0.5012 Average %RPD (3)36 .3 "'39 .7 m86.6 ~ 13 '~"379 "s ~384 "'864 "'382 1 .4 I %RSD NA "'0130 P)1 .36 NA P)0 .121 ")1 .17 c')96 .6 "'0,143 43 Average %Recovery -~ .., %RSD 5.1 ~'130 121 m142 -~"131 8 .4 SM EP1VIRONMENTAL LA60RATORY PAGF 16 OF 31 3M ENVIRONMENTAL LABORATORY REPORT N0 . E09-0053 Table 8 Continued. PFOS 'O PFOS 3M L1MS ID Sam e Descri 'on mass Conc. n (I)Conc. n %.Rec Conc. n Conc. n %Rec E09-0053-009 MissR_pooi2-SM8-3 0.4963 11 .0 E09-0053-009 Dup 0.4975 12 .2 E09-0053-009 LMS 0.47 84 26.8 Average %RPD E09-0053-010 MissR_pool2 SMB 4 0 .5107 13.4 E09-0053-010 Dup 0 .4952 12 .1 E09-0053-010 LMS 0 .4839 26 .6 ---- - Average %RPD 111 123 280 _ 117 10 531 122 _ 275 -127 7.1 NA NA 103 NA NA _ 94 .4 0.106 1 .07 106 0.116 1 .17 116 0 .116 1 .21 116 ----------------- ------ Average %Recovery 112 %RSD 5.1 0 .119 1 .17 119 0.102 1 .03 102 0.124 _ 1.28 __ 12_ 4 Average %.RecoverY 115 %RSD 10 E09-0053-011 E09-0053-011 Dup E09-0053-011 LMS MissR_pool2 SMB 5 0 4926 0 .4861 0 5148 Average 13 .9 14 .6 -27 .6 141 NA 0.114 1 .16 114 150 NA 0.120 1 .23 120 268 83 .0 0.122 1__....1_.8..-------12-2-- 146 Average %Recovery 118 %RPD E09-0053-012 MissR_poof2_BKS_1 0 .4749 5.27 E09-0053-012 Dup 0 .5146 561 E09-0053-012 LMS 0 .5028 22 .5 Average ~RPD E09-0053-013 MissR_poo12 BGS-1 G .5058 ~ 15 .7 E09-0053-013 Dup 04887 16 .4 E09-0053-013 LMS_- . _ ...._ ._.. .. . . . . .__. .. . . ...__.-..-.. . ....___ ._. ._ ._.._. .. ._. Average 6.2 55 .5 NA 54.5 NA _224 _ 112 55.0 -- - ----- 1 .8 155 NA 168 NA 343 161 123_ -- /.RSD 3.5 0.0995 1 .05 99 .1 0.105 1 .02 105 0.116 _1 .1 5 _1_16_ Aver-age %.Recovery 108 _ Y*RSD 7.9 0.110 1 .09 110 0.105 1 .07 105 -0-.-1-08--...----1-.-0-5___._ ..._... .-1--0-8--- Average %Recovery 107 E09-0053-014 E09-0053-014 Dup E09-0053-014 LMS E09-0053-015 E09-0053-015 Dup E09-0053-015 LMS E09-0053-016 E09-0053-016 Dup _E09-0053-016 LM.S. MissR_poo12 BGS-2 MissR_poo12_BOS_3 MissR-poo12 BOS-4 .... . . . ... _._. . . . ...... ioRPD 7.8 %RSD 23 0.5237 04809 0.5201 11 .5 110 9 .61 100 26_0 _--., 250 NA NA 99~3 ., ._ 0.123 0 .109 .0.113-._. . 1 .17 1 .13 1 . 0_9 123 109 113_ Average 91,RPD 105 Average Y.Recovery 115 9.4 %RSD 6.3 0.5118 19 .2 188 NA 0 .108 1 .06 108 0.5022 16 .8 167 NA 0 .112 1 .12 112 0.5098 37.4 367 127 0.117 1 .15 117 Average 177 Average %Recovery 112 %RPD 11 %.RSD 4.0 0.5024 "'33 .3 '3~331 NA "'0.122 P~1 .21 122 0 .5096 P)34 .7 (')340 NA ('10 .118 1,11 .16 118 00.;5124......_ . . ._-~'83 .7.--.. .. . . . .. .._ .rn817 .... . . . . _. -_ ~~98 :5-. . . .- a10.163._ ...__. . 1. '.1_1.,_.._5...9. . . ...... -. ._ __~... .'-1--6- .2..._ . Average m336 Average %Recovery c'134 %RPD 2.7 %RSD 19 3M ENVIRONMENTAL LABORATORY PAGE 17 OF 31 3M ENVIRONMENTAL LABORATORY REPORT NO . E09-0053 Table 8 Continued. - -- PFOS - - 'O PFOS I 3M UMS JD sample Descrl tion ma- Conc. n L (f)Conc. n %Rec Conc . ngimL) Conc . (ncyg) E09-0053-017 MissRj)ool2 BGS-5 0.5005 12 .3 E09-0053-017 Dup 0 .5146 12 .4 E09-0053-017. LMS. . . . . .,.., .. . . . . .--_ - . ,_,- . . .. . . . ..._ . . 0.5052 28_0 Average 123 NA 120 NA . 277_ _. -, ... . .. . . . . .1._ 0. . ..3--122 0 .116 1 .16 0 .104 1 .01 0 .118 1 .17 - Average %Recovery %RPD Zo %RSD E09-0053-018 MissR_pool3 WE_1 0.5021 9.78 97 .4 KA 0 .102 1 .02 E09-0053-018 Dup 0.5091 9.72 95 .5 NA 0 .102 1 .00 -E-09--...0. .0..5...3._-_..0...1--8---L---M- S. .._ ._ . .------ ----------...... --_. . . . .------_0-..-5..1._6..8 27 .1 262 113 0 117 1 .13 Average 96 .4 Average %Recovery %RPD _ 2.0 %RSD E09-0053-019 E09-0053-019 Dup MissR_pool3 WE_2 .....E- .-0. ...9_._-. .0--0--5-3-----019 LMS .. ....---- . . .------ .. .... .-----.._ .___. . ._ . 0.5233 8.20 0 .5150 8 .10 0:4892--.. . ._ ._ 24 :9 Average 78 .3 NA 78 .6 NA 254... . .__ - .113 78.5 0.115 1 .10 0.105 1 .02 0.119 1 .22 Average %Recovery r6RPD 0.37 %RSD E09-0053-020 MissR_pool3 WE 3 0.5075 9 .89 E09-0053-020 Dup 0 .5040 8 .30 E09-0053-020--L.M-S-._ ._...._ . . . ._.._....... . . . .. ... . ... ..___ .. . . .._ . . .___ . .._ _ . 0. . . ... 5.,1.7_2._-_ .._ ._. .. . _.. .2. 8.. :. 7. ., - Average 97 .4 NA 0 .130 1 .28 82 .3 NA 0 .106 1 .05 _ . . .. .___2_ 7.._7. . ......._-_- --_-.--1--2---8--__ ... . . . ..0-.-112-2- _ 1 .18 89 .9 Average %Recovery %RPD 17 %RSD E09-0053-021 E09-0053-021 Dup E09-0053-021 LMS MissR_pool3-WE 4 0.5093 I 5.03 0.5144 5.38 _-----------.0_-.-5.1-6_8---- ~ 21 .0 Average 49 .4 NA 0110 1 08 52 .3 NA 0 .100 0 .972 ---2. 0. .3._.. .---- ---. . . 1.-0--4... . . .. . .-0_..I ..1-0- ..5.--- . -.-... --- 1-.02 50.8 Average %Recovery %RPD 5.7 %RSD E09-0053-022 E09-0053-022 Dup _ E09-0053-022 LMS MissR_pool3 WE 5 0.5152 0.4847 0;4779 Average 6.62 6.60 -23 .7 64 .2 68 .1 . .__.._2~ 66.2 NA ~ 0.100 0 .970 NA 0 .105 1 .08 114 - _0 .123 1 .29 Average %Recovery %RPD 5.8 %RSD E09-0053-023 MissR_pool3._NOP 1 0.4741 4 .72 49 .8 NA 0 .108 1 .14 E09-0053-023 Dup 0,5087 5 .21 51 .2 NA 0.108 1 .06 E09-0053-023 LMS . -- ._ ._ ,-. ., ._ .. ._ .._ . . ._ . . _ . . . . . .__ . .0..5024 ,. , ., -...21. .7 216 109 0.132 1 .31 Average 50.5 + Average /Recovery %RPD 2.8 ' %RSD E09-0053-024 s MissR_pool3-SMB_1 04791 2.83 29 .5 NA 0 .110 1 .15 E09-0053-024 Dup 05276 3.15 29 .9 NA 0 .102 0 967 E09-0053-024 LMS.,.. _,.- . .-_ . 0.5115 _ 1.8 .2 178 100 0.114 1 .11 Average 29 .7 Average 5/.Recoveery %RPD - ~ 1.1 %RSD -- %Rec 116 104 118 112 6.7 102 102 117_-_-. 107 8.1 115 105 119 113 6.4 130 106 122 119 10 110 100 105 105 4.8 100 105 123 109 11 108 108 m132 116 12 110 102 114 108 5 .6__~ 3M ENVIRONMENTAL LABORATORY PAGE 18 OF 31 3M ENVIRONMENTAL LABORATORY REPORT NO . E09-0053 Table 8 Continued. PFOS 'O PFOS 3M UMS !D Sample Descd Bon Mass Conc. n L (1)Conc. n %Rec E09-0053-025 MissR_.poo13 SMB 2 0.4868 10 .0 103 NA E09-0053-025 Dup 0.5146 11 .1 108 NA E09-0053-025 LMS _~ . ._-_---- 0.5212 -- 25 .9 248-- 98 .2 ------ ^- Average 105 --- %RPD 4.9 E09-0053-026 MissRj)ool3_SMB 3 0.5004 4 .88 48.8 NA E09-0053-026 Dup 0.4942 4.77 48.3 . NA E09-0053-026 LMS_._...----- . ._ .4928 -------------------0 20:0 Average 203-- -- .._- ._..1_0_0-. 48.5 %RPD 1 .0 E09-0053-027 MissR_pool3 SMB 4 0.4802 6.05 63 .0 NA E09-0053-027 Dup 0.4978 6.66 66.9 NA -E09-0053-027 .LMS-..,.,., . .-.._ . . ._ ..__..__ .-_.... 0_5202 --- - _20;2--,-----l194 88 .5 Average 64.9 Conc. n Conc. n %Rec 0 .105 1 .08 105 0 .125 1 .21 125 0.122 _1.17 - 122 Average %Recovery 117 %RSD 9.2 0 .108 1 .08 108 0.105 1 .06 105 0.12 1 .22------- ---120 Average %Recovery 111 %RSD 7.2 0.112 1 .17 112 0.115 1 .16 115 -0 .118 1 .13 ~.-1- 18-. . Average %Recovery 115 oRPD 6.0 %RSD 26 E09-0053-028 MissR_poo13 SMB_5 0.5170 5.69 55 .0 NA 0.108 1 .04 108 E09-0053-028 Dup 0.5016 5.35 53 .3 NA 0.114 1 .14 114 --E09-0053-028 LMS, . .... .----,-, - - 0.5015 20.6 _ 205 100 _ 0.12 __1 .20 __12_0 Average 54 .2 -- ------- - Average %Recovery 114 %RPD 3 .1 %RSD 5.3 E09-0053-029 MissR_pool3 BGS_1 0.4922 7 .55 76 .7 NA 0.113 1 .15 113 E09-0053-029 Dup U .5236 7.62 72 .8 NA 0.0988 0 .943 98.4 E09-0053-029 LMS-~-----_`- 0.5289 - 21._3_-` 201 - 88.2 0 .112 112 -------- Average 74.7 Average %Recovery 108 %RPD 5.3 %RSD 7.3 E09-0053-030 MissR_pool3 BGS_2 0.5234 16 .8 160 MA 0 .104 0.994 104 E09-0053-030 Dup 0 .5010 16 .5 165 NA 0 .115 1 .15 115 E09-0053-030 LMS 0.4780 Average 28 .3 -----2-9--6----. ._... .---8--4 .0 163 0.122 ----.. .--1 -.2-8-..--------1.2-2---- Average %Recovery 113 %RPD 26 %RSD 8.0 E09-0053-031 MissR_pool3 BGS 3 0.4868 13 .4 138 NA 0 .119 1 .22 119 E09-0053-031 Dup 0.5055 12 .9 128 NA 0.105 1 .04 105 E09-0053-031 LMS 0.4815 _ Average . --25- .-8--. . .__----2-6-8------.... .--8-5--.-8---... . .--0-.-1-2-2--.._ ..o----1 -.-27-----._. ._..--1-2-2---- . . 133 Average /oRecovery 115 %RPD 7.6 %RSD 7.9 E09-0053-032 MissR_pool3 BGS 4 0.5164 23 .0 223 E09-0053-032 Dup 0.4841 21 .2 219 E09-0053-032 LMS- - , _ ._ . __ . . ...._ ._. ....... _.___ 0.5015 ., - . . . 34.5_ . 344 Average 221 NA 0.105 1 .02 105 NA 0.115 1 .19 115 81 .3 0.124 1 .24 . . .. .__._ .----_1...2. . . .4.._..__.. Average %Recovery 114 %RPD 1 .7 %RSD 8.3 3M ENVIRONMENTAL LABORATORY PAGE 19 OF 31 p. 21 3M ENVIRONMENTAL LABORATORY REPORT NO. E09-0053 Table 8 Continued. - PFOS 'e0 PFOS 3M UMS !D E09-0053-033 E09-0053-033 Dup E09-0053-033 LMS Sam e Deny Bon MissR_poo13 BGS_5 Mass 0 .5092 0.5071 0 .4998 Average Conc. n mL 12 .8 13 .5 26 .0 ItiConc. n 126 133 260 129 I %Rec NA NA . 86 :0- . ~ ~ Conc . n mL Conc. n 0117 1 .15 0.121 1 .19 0 .120 1 .20 Average %Recovery %Rec 117 121 120 119 %RPD 5.7 . %RSD 1.7 (1) Unless otherwise specified, the recovery of associated laboratory matrix spike (LMS) was within 10000t3300/%~ which meets ETS 8-45 .0 method acceptance criteria . Sample results are considered accurate to within the overall analytical method uncertainty 100t15% (PFOS). See Section 4.9 Determination ofAnalylical Method Uncertaintyfor more information . Sample results are reported to three significant figures and %RPD to two significant flgures. Additional significant figures were used to calculate the average, %RPD, and %recovery. Values in the rawdata may vary slightly due to rounding . (2) (3) Surrogate recovery exceeded 100t30% Reported concentrations forthese samplrs are from a 1:10 post-extraction dilution of the final extract. These two samples were re-prepared and analyzed in Batch 4 with z -500 ng/g LMS. The results from the original analysis were not reported as the l1VIS concentration (-150 nglg) was less than half the endogenous sample concentration and consequently, the accuracy of the sample concentrations could not be assessed . Figure 1. Area Percentages of Individual PFOS Transitions : Samples -001 through -018. 90 .00- 80 .00 ---r-- 0 +i 70 . 00 don 60 .00 50 .00 ad 40 .00 30 .00 Q d 20 .00 I " % 499>1301 I " % 499>99 i l " %0 499>BOjI 10 .00 > a 0 .00 g O) O - N M V LO (0 r-- W .N O~ O Qc) O OM 0 2 IL Lo Zo U-) O 9MLo 4CU-l)) O 4 U-) O O Lo U-) LO O Q Q O ~ q Lo LO M M un "') 4 Lo M Lo Q O C~ O C~ O O O OO O O q O O G O O O O O, Q O 4 O O Q 4 Q O 4 O C? O C? O O C07 O O O O O O O WW WW WW W W aJ W W ~ ~a1 C> W C) LJJ Z W l11 W U c > 0 3iV1 Ei"!`iRONMENTAL LABORATORY PAGE 20 OF 31 3M ENVIRONMENTAL LABORATORY REPORT NO . E09-0053 Figure 2. Area Percentages of Individual PFOS Transitions: Samples -019 through -033 . 90 .00 0 80.00 ~ 70 .00 -I aay> 60 .00 cy .7V.VV "I a 40 .00- 30 .00- Q d m 20 .00 - 10.00 > 0 .00 - c~ N N q N O N N N N N N Q N Q (4'~ cq'7 cN7 O eqM7 C 994 4 O o `n o 0 0 `n `" o o `~ o 0 0 0 L0 CL 4 S 4 C? 9 $ $ 9 q $ ~ ~ 4 $ a 0w 0w 0w 0w 0w 0w 0w 0w 0w w 0w 0w 0w 0w 0w Z U c > 0 0 V) 499>130 , 499>99 n 499>80 The PFOS sample results presented in Table 1 and Table 8 are considered accurate within the overall method uncertainty of 100t15%. When c;ompared to the solvent standards used for quanitation, area count ratios of the individual PFOS transitions (499>80, 499>99, and 499>130) were conserved in extracted samples. The accuracy (% recovey) and precision (%RSD) of 3M ECF (branched) PFOS laboratory control spikes quantitated against the predominantly linear reference standard demonstrated that the isomers produce no discernable quantitative bias . Results for remaining target analytes will be in a phase II report. e2fkerenc' s." [1] J. P. Benskin, M. Bataineh, J.W. Martin, Simultaneous Characterization of Perfluoroalkyl Carboxylate, Sulfonate, and Sulfonamide Isomers by Liquid Chromatography-Tandem Mass Spectrometry, Anal. Chem . 79 (2007) 6455-6464. 3M ENVIRONMENTAL LABORATORY PAGE 21 OF 31 3M ~NVIRONVIENTAL LABORATORY REPORT NO. E09-UQ53 All remaining sample and associated project data (hardcopy and electronic) will be archived according to 3M Environmental Laboratory standard operating procedures 9.1 3M Environmental Laboratory Michelle D. Malinsky, Ph .D Research Specialist 9 .2 Pace Lab Ops Jonathan Steege Lab Analyst II 3h1 E:N't'IRONNF_NiAL LA80R,4TOR'Y' PAGE 22 OF 31 3M ENVIRONMENTAL LABORATORY REPORT NO. E09-0053 Mich~lle D. Malinsky, Ph .D, M-~esearcl(~Ocialist, Principal Analytical (investigator Date William K. Reagen, Ph.D., Environmental Laboratory Management /~`c.i~ aUOI~ Dat, e Cliffton"li'. Ja6by, Ph .D ., M Technical Reviewer 1,4,-C -'~?9 Date The 3M Environmental Laboratory's Quality Assuranoe Unit has audited the data and report for this project. QAI Represendive Date 3M ENVIRONMENTAL LABORATORY PAGE 23 OF 31 O Ow J Z J ~ Za Z w M p. 25 ~OOU~ ~ pj O7 0 ~ C p o~ o i1~ > r ~0, ~ 1q G O O U O O O T ~ N U - --> I~ , N N N" (O Q~ G U O~ U ,\ U L U E N O M O ~ 000 ~ 0o1 y ~ C N '- M N O ~G O O O O U U DC E G O N o M o N $ o ~ N i~ ~ ~mc~ n VG o00000 0 b a O ~ ~U e J O O o 0 0UOO` N 0 n 0 ~p O? U rD a0 cn u7 C1 N GO O ~ ~ ~O ~O r O ~ \ 00 I I , ~ N lo o o (N a) a' 0 a o "a 0 N W v ! 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