Document O3g9N1Y3zLgJB4mxvvN9o6Raj

p' 3M /\-(tHP.6~ $S~7 3M ENVIRONMENTAL LABORATORY PROJECT NO.EOS-0442 Analytical Report Definitive Study: Quantitative Screening of Commercial Lots of Human Serum Obtained in 2004 for Endogenous Fluorochemicals Using Solid Phase Extraction and Liquid Chromatography/Tandem Mass Spectrometry Laboratory Request Number: E05-0442 Testing Laboratory 3M Environmental Health and Safety Operations 3M Environmental Laboratory 3M Environmental Laboratory Project Lead Mark E. Eilefeon Building 2-3E-09 PO Box 33331 Saint Paul, MN 55133-3331 Phone: 651-778-5405 Testing Facility Management W illia m K . R e a g e n , P h .D . Building 2-3E-09 PO Box 33331 Saint Paul, MN 55133-3331 Phone: 651-778-6565 C3 cn CO> rn o r-.> CO -3* o 03 Requestor Dale Bacon Building 2-3E-09 PO Box 33331 Saint Paul, MN 55133-3331 Phone: 651-778-4736 Page 1 of 19 3M ENVIRONMENTAL LABORATORY PROJECT NO.EQS 0442 3M Environmental Laboratory Analytical Report Project Number, E5-0442 1 Introduction i Summary The following analytical report presents data from commercially available normal pooled human serum that were purchased by the 3M Environmental Laboratory and subsequently screened for endogenous fluorochemicals. Thirty-six separate lots o f pooled normal human serum were extracted using a solid phase extraction technique and screened for sixteen different fluorochemicals using liquid chromatography/tandem mass spectrometry (LC/MS/MS). Serum samples were received from various suppliers between 9/9/04 and 12/2/04. Samples were extracted on 5/11 /2005 and analyzed 5/23/2005. Heptafluorobutyric acid (PFBA) was included in the original list o f sixteen target analytes. However, due to instability in the methanol extract, analytical data for PFBA will not be included in this report. Instead, a separate study will be conducted including the re-extraction o f the thirty-six lots o f commercial serum with acetonitrile and analysis using a newly developed method that provides isomer resolution. Those results will be reported in a separate report. Sample results are summarized in Tables 1 and 2. The average values reported represent samples in which there was detectable analyte. Sample values that were less than the limit of quantitation were not included in the average value calculations. Please refer to Section 5 for individual sample results. Page 2 of 19 mtm -J * 3M ENVIRONMENTAL LABORATORY PROJECT NO.05-0442 Table 1. Summary of Perfluorinated Acid Results Analyte Mean Median High Low Standard Deviation Number of Samples with Detectable Analyte NFPA PFHA TDHA PFOA (ng/mL) (ng/mL) (ng/mL) (ng/mL) C9 Acid (ng/mL) C10ACID (ng/mL) C11 C12 ACID ACID (ng/mL) (ng/mL) 1.04 o.ouV 0.151 ' 2.98 0.813 0.282 0.185 0.0246 1.05 2.11 0.327 0.378 0.00838 0.0323 0.00633 0.00822 0.118 0.474 0.0232 0.111 3.21 0.737 5.72 1,97 0.359 0.0950 1.27 . 0.445 0.281 0.599 0.0274 0.137 0.134 0.734 0.0152 0.176 0.0197 0.0833 0.0108 0.0149 3S/36 9/36 33/36 36/36 36/36 36/36 36/36 30/36 Table 2. Summary of Other,Perflu.oripa.ted and Highly Fluorinated Analyte Results Analyte Mean Median High Low Standard Deviation Number of Samples with Detectable Analyte FBSA (ng/mL) NA1 NA' FOSA PFBS (ng/mL) _ (ng/mL) 0.149 0.0165 0.124 0.0101 0.00542 - 0.446 0:0840 0.00542 0.0376 0.00545 NA1 0.0876 . 0.0169 PFHS PFOS THPFOS THPFDS (ng/mL) (ng/mL) (ng/mL) (ng/mL) 2.55 14.9 NA2 0.0442 2.85 15.5 NA2 0.0366 4.18 27.0 NA2 0.126 0.606 2.03 NA2 0.00772 1.07 5.94 NA2 0.0227 1/36 36/36 33/36 36/38 36/36 0/36 36/36 NA2 = Not Applicable because this analyte was not detected in any of the samples. Page 3 p f 19 3M ENVIRONMENTAL LABORATORY PROJECT N0.E05-0442 2 Reference Substances Table 3. Reference Substances Name Nonafluoropentanoic acid Synonym or Acronym NFPA (CS Acid) Standard Source Material Vendor Traceability Number Purity * Aldrich TCR-333 (00017-945) 96,5% Additional Correction Factor (K* salt or H*) 0.996 Periluorohexanoic acid PFHA (C6 Acid) Oakwood Products TCR-673 97.7% 0.997 Tridecafluoroheptanoic acid TDHA (C7 Acid) Aldrich TCR-267 98.2% 0.997 Perfluorooctanoic acid PFOA (C8 Acid) 3M TCR-123 (99030-030) 95.0% 0.958 Hcptadecafluorononanoic acid C9 Acid Oakwood Products TCR-618 98.0% 0.998 Nonadecafluorodecanoic acid CIO Acid Oakwood Products TCR-36 98.01% 0.998 Peril uoroundecanoic acid Perfluorododecanoic acid C U Acid C l 2 Acid Oakwood Products Oakwood Products TCR-619 TCR-37 (SD037) 96.4% 99.65% 0.998 0.998 Perfluorobutancsulfonam ide FBSA 3M TCR-314 (00017-026) unknown None Perfiuorooctanesulfcm amide FOSA 3M TCR-280 (99131-038) 98.94% None Perfiuorobutane sulfonate PFBS 3M T C R -116 (99030-023) 96.7% 0.8844 Pcrfluorohcxane sulfonate PFHS 3M TCR-83 (SE036) 98.6% 0.911 Perfluorooctane sulfonate PFOS 3M TCR-696 86.4% 1H, 1 H .2 H .2 H , pcrfluorooctanesulfonic acid THPFOS SynQuest Labs TCR-343 (00017-055) 93.5% 1H, 1H, 2 H ,2H , perfluorodecane sulfonate THPFDS Pace/iM TCR-627 94.7% Perfluorooctanoic acid, isotopicatly labeled Perfluorooctane sulfonate, ammonium salt, isotopically labeled P F O A [l,2 -|3C) PFOS [,llOj] Perkin Rimer RTI International TCR-744 TCR-756 >97% 100 ug/mL Standard * Reference substance purities have not been corrected for impurities due to shorter drain homulogues which cannot be distinguished by NMR analysis. 0.927 0.998 0.998 None None . NMR results indicate that the PFOS reference substance consists o f 30.8% branched isomer, and the PFOA reference substance consists of 22.1% branched isomer. These values were used for the determination of the isomer ratios in the serum samples (presented in Table 10). Page 4 of 19 t 3M ENVIRONMENTAL LABORATORY PROJECT NO,EOS-0442 3 Sample Preparation The samples for this study were prepared as follows: l . The serum samples were allowed to thaw and warm to room temperature. 2. Four milliliters o f serum (or water in the case of method blanks) were added to a 15 mL screw-capped polypropylene centrifuge tube. 3. Matrix spikes and method blanks were prepared by spiking the serum (or water) with the analytes o f interest. In addition, l3C PFOA and l802 PFOS were added as surrogates to all serum samples, calibration standards, matrix blanks, and method blanks at concentrations of 1.04 and 0.950 ng/mL serum respectively to evaluate extraction method performance. 4. Ten milliliters o f acetonitrile were added to the serum and the centrifuge tubes were capped and vortex mixed, allowing the serum proteins to precipitate. 5. The tubes were centrifuged at approximately 3500 rpm for 20 minutes to pellet the precipitate. 6. After removal from the centrifuge, the supernatant was transferred to a bottle containing 70 mL of water. 7. The samples were passed through a preconditioned SPE cartridge and eluted with 2 mL o f methanol. Quantitation was accomplished using high performance liquid chromatography tandem mass spectrometry. The following parameters were used: Liquid Chrom atograph: Hewlett-Packard Series 1100 Liquid Chromatograph system Analytical column: Keystone BetasilTM Cj* 2.1x100mm, 5pm particle size Column temperature: 30 C Stop Time: 20.0 minutes Flow rate: 300 pL/min Injection volume: 5 pL Mobile phase components: Solvent A: 2.0 mM ammonium acetate in ASTM Type I water Solvent B: HPLC Grade Methanol Solvent Gradient: Time 0.00 1.00 14.50 15.50 16.50 20.00 20% 20% 90% 90% 20% 20% Page 5 of 19 3M ENVIRONMENTAL LABORATORY PROJECT NO.EOS-0442 Mass Spectrometer: MDX Sciex API 4000 Q-Trap Mass Spectrometer Detector Software: AnalystTM Version 1.4 Ionization Mode: Electrospray Negative Temperature: 450 C GS1: 35 GS2: 45 Entrance Potential:-10 Analysis Type: Multiple Reaction Monitoring (MRM) Table 4. Acquisition Parameters Comoound N F P A (C 5 A rid ) P F H A (C a A c id ) T D H A (C r A c id ) P F O A (C 8 A c id ) C 0 A cid C 10A cid C t1 A cid G f2 A cid FBSA FOSA PFBS PFHS PFOS THPFOS THPFDS P F O A [1 ,2 -13C ] P F O S [18O d Q1 Mass lamu) 262.9 313.0 362.9 41^0 463.0 512.9 563.0 613.0 297.9 497.9 298.9 398.9 498.9 426.9 526.9 414.9 503.0 <23 Mass laitiul 219.0 268.7,118.9 318.7,168.8,118.9 368.9,219.0,169.0 418.7,168.9,218.9 468.8,218.9.269,1 518.7,268.9,218.8 568.7.168.9.318.7 78.0 77.9 9 8 9,79 .9 98.9,80.0 8 0 .0 ,9 8 .9 ,1 3 0 .0 4 0 8 .8 ,8 0 .9 506,7,81.0 369.8 103.0, 84.0 Declusterina Potential -30 45 -35 -45 -45 -35 45 -50 -55 -90 -65 -95 -120 -70 -70 -40 -100 When multiple transitions were monitored, the total ion current (TIC) was used to quantify each analyte, thereby providing greater sensitivity than individual Q3 ion measurements. 4 Analysis 4.1 Calibration 4.1.1 Calibration Curve An extracted calibration curve was formed by spiking rabbit serum at prescribed levels and extracting the curve in the same manner as the samples. Stock solutions were individually prepared from neat fluorochemicals and diluted in methanol. Samples were quantified versus a 15-point extracted calibration curve. Page 6 of 19 3M ENVIRONMENTAL LABORATORY PROJECT NO.E05-0442 The regression analysis was performed using a quadratic curve fit weighted 1/x. Calibration standards werewithin 25% (30% for the Lower Limit of Quantitation (LLOQ)) with a coefficient of determination >0.990. Low-level calibration standards which were not within 30% were not included in the calibration curve. Calibration standards that were not greater than 2x the method blanks were not included in the calibration curve, except where noted. 4 .1 2 Continuing Calibration Verification (CCV) A Continuing Calibration Verification (CCV) was run at least once every 10 samples, bracketing the samples with the calibration curve. The CCV recovery was within 25% for all analytes. 4.1.3 System Suitability Five system suitability standards were analyzed before the initial calibration curve and after tire last CCV. These samples had area counts with a %RSD of <5% and a retention time %RSD of2% when evaluated independently except for the following instances: The system suitability samples for each o f the perfluorinated acids and FOSA failed. In each case the %RSD of the retention times was well within tolerance limits, but the area counts varied by as much as 20.3%. The failure of every perfluorinated add tested suggests the presence of a systematic effect on that class of analytes. This should be kept in mind when evaluating the data presented in this report. 4.1.4 Lower Limit of Quantitation (LLOQ) The LLOQ for each analyte was equal to the nominal concentration o f the lowest standard in the calibration curve that met accuracy requirements o f 70% -130% and that had at least twice as many area counts as the method blanks. 4 .2 B lanks 4.2.1 Method Blanks Method blanks consisted of aliquots of Milli-Q water extracted in the same manner as the samples. All of the method blanks in this studyhad area counts < 50% of the LLOQ for their respective analytes with the following exceptions: PFHA: Two out ofthree method blanks was greater than 50% o f the LLOQ; none were above the LLOQ. C7 Acid: One of three method blanks had area counts greater than 50% of the LLOQ; none were above the LLOQ. C11 Acid: One o f three method blanks had area counts greater than 50% of the LLOQ; none were above the LLOQ FOSA: Two out of three method blanks were greater than 50% o f the LLOQ; none were above the LLOQ. Page 7 o f 19 3M ENVIRONMENTAL LABORATORY PROJECT NO.E05-0442 PFBS: Two method blanks had area coiihts greater than 50% o f the LLOQ; one showed anomalous contamination and was above the LLQQ. 4 2 2 Matrix Blanks Matrix blanks consisted of aliquots ofblank rabbit serum extracted in the same manner as the samples. Three matrix blanks were prepared and analyzed with the samples. Matrix blanks containing endogenous levels o f analyte that were above die LLOQ are detailed below. Table 5. Endogenous Analyte in Rabbit Serum Sample ID TDHA C11 ACID PFBS THPFOS THPFDS LLOQ (ng/mL) 0.00504 0.00992 0.00506 0.00520 0.00503 MxBIk 050511-1 0.0065 0.0196 0.00941 0.0260* 0.0146 MxBIk 050511-2 0.00252* 0.0138 0.00729 0.0260* 0.0131 MxBIk 050511-3 0.0102 0.0169 0.00774 0.0249 0.0137 Average 0.00641 0.0168 0.00815 0.0256 0.0138 ` Please refer 10 the note to file included in the raw data describing the calculation o f the endogenous levels. The average endogenous values were used to correct the matrix spike response. 4 2 3 Solvent Blanks Methanol blanks were analyzed throughout the analytical run. Analyte response was less than lA the LLOQ in all cases. Surrogate PFOA [1,2-13C] and PFOS [1802] were added as surrogates to all serum samples, calibration standards, matrix blanks, and method blanks to evaluate the extraction method performance. All PFOA [1,2-' 3C] surrogate recoveries were within 3 0%, but two of the PFOS [1S02] recoveries were outside +30%, extending the surrogate recovery range to +/-34%. Specific surrogate recovery data is presented below. Page 8 of 19 3M ENVIRONMENTAL LABORATORY PROJECT NO.E05-0442 Table 6. Surrogate Recovery wimmiSm Sample ID PFOA [1,2-1SC] PFOS [1sOJ Recovery, % Recovery, % MxBlk_0505t1-1 MxBlkJ)50511-2 MdBlk_050511-1 MdBlk_050511-2 LS_Q505115-1 LSJJ505115-2 LS,,050S 115-3 HS_0505115-1 H S_0505115-2 HSJJ505115-3 TN-A-6840 TN-A-6843 TN-A-6847 TN-A-6934 TN-A-6935 TN-A-6944 TN-A-6959 TN-A-6960 TN-A-6967 TN-A-6973 TN-A-B975 TN-A-6977 TN-A-6981 110% 102% 80% 99% 114% 109% , 98% 85% 120% 90% 80% 93% 78% 102% 89% 80% 85% 78% 91% 76% 84% 62% 83% m% 98% 112% 103% 103% 94% 98% 95% 98% 88% 87% 82% 79% 72% 82% 79% 80% 66% 85% 76% 82% 73% 67% Sample ID PFOA [1,2-1*C] Recovery, % PFOS [18OJ Recovery, % TN-A-5992 TN-A-6994 TN-A-6995 TN-A-6997 TN-A-7000 TN-A-7006 TN-A-6938 TN-A-S939 TN-A-6941 TN-A-6950 TN-A-6953 TN-A-6955 TN-A-6962 TN-A-S964 TN-A-6966 TN-A-6970 TN-A-6972 TN-A-6978 TN-A-6979 TN-A-6982 TN-A-6984 TN-A-6966 TN-A-6988 : . 94% 87% 83% 84% 78% 91% 92% 78% 88% 96% 76% 85% 84% 89% 83% 84% 73% 74% 92% 113% 86% 81% 85% 83% 77% 74% 75% 72% 84% 76% 88% 82% 85% 81% 81% 85% 86% 75% 84% 75% 80% 78% 87% 86% 83% 83% 4 .4 L aboratory C ontrol Spikes (LCS) Six Laboratory Control Spikes (LCS) were prepared in rabbit serum and analyzed with the samples. Results were corrected for endogenous levels o f analyte in the matrix (where appropriate), and are presented below. Page 9 of 19 3M ENVIRONMENTAL LABORATORY PROJECT NO. E05-0442 Table 7. Matrix Spike Recoveries A nalyte NFPA S a m p le ID - N om inal O bserved Recovery, % C one., ng/m L C one., ng/m L LS 0505116-1 LS 0505115-2 LS 0505115-3 HS 0505115-1 HS 0505115-2 HS 0505115-3 0.306 0.306 0.309 3.06 3.06 3.06 0.517 0.373 0,336 2.89 3.48 3.55 159% 122% 109% 94% 114% 116% % RSD 23% 11% PFHA LS.,,0505115-1 LS 0505115-2 LS 0505115-3 HS 0505115-1 HS 0505115-2 HS 0505115-3 0.303 0.303 0.303 3.03 3.03 3.03 0.699 0.281 0.224 3.43 2.79 3.09 231% 93% 74% 113% 92% 102% 65% 10% Page 10 of 19 3M ENVIRONMENTAL LABORATORY PROJECT NO.E05-0442 Table 7. Matrix Spike Recoveries (continued) PFOA LS 0505115-1 LS 0505115-2 LS 0505115-3 HSI05055-1 ' HS 0505115-2 HS_0505115-3 0.297 0.297 0.297 2.97 2.97 2.97 0.235 0.129 0.136 2.46 2.82 2.7 70% 43% 4B% 83% 95% 01% C9 Acid C10AC1D LS 0505115-1 LS 0505115-2 LS 0505115-3 H S_0505115-1 HS 0505115-2 HS 0505115-3 LS 0505115-1. LS 0505115-2 LS 0505115-3 HS 0505115-1 HS 0505115-2 HS .0506115-3 0.300 0.300 0.300 3.00 3.00 3.00 0.311 0.311 0.311 3.11 3.11 3.11 0.285 0.237 0.31 2.14 3.3 3.04 0.291 0.323 0.323 3.30 3.16 3.06 95% 79% 103% 71% 101% 101% : 94% 104% 104% 106% 102% 98% C12 ACID LS 0505115-1 LS 0505115-2 LS 0505115-3 HS 0505115-1 HS 0505115-2 HS 0505115-3 0.30 0.30 0.30 2.99 2.99 2.99 0.323 0.371 0.341 2.91 3.65 3.56 108% 124% 114% 97% 122% 119% FBSA LS 0505115-1 LS 0505115-2 LSJ3505115-3 HS 0505115-1 HS 0505115-2 HS 0505115-3 0.300 0300 0.300 3.00 3.00 3.00 0.305 0354 0.318 3.00 3.25 3.51 102% 118% 106% 100% 108% 117% 36% 5.9% 13% 19% 5.9% 3.8% 7.0% 12% 7.8% 7.6% Page 11 of 19 Table 7. Matrix Spike Recoveries (continued) 3M ENVIRONMENTAL LABORATORY PROJECT NO.E05-0442 A nalyte FOSA PFHS PFOS S a m p le ID LS 0505115-1 LS 0505115-2 LS 0505115-3 HS 0505115-1 HS .0505115-2 HS 0505115-3 N om inal C one., ng/m L O b serv ed C one., R ecovery, % ' ng/m L 0.299 0.299 0.299 2.99 2.99 2.99 0.370 0.274 0.251 2.79 3.45 2.36 124% 92% 84% 93% 115% 79% LS 0505115-1 LS 0505115-2 LS 0505115-3 HS 0505115-1 HS 0505115-2 HS 0505115*3 0.315 0.315 0.315 3.15 3.15 3.15 0.305 0.302 0.317 3.12 2.35 2.90 97% 96% 101% 99% 90% 92% Q C 0 50202-001 Q C 0 50202-002 QC050202-003 QC050202-004 QC050202-005 QC050202-006 0.303 0.30S 0.303 3.08 3.08 3.08 0.195 0.160 0.230 3.07 3.06 2.52 63% 52% 75% 100% 100% 82% % RSD . 21% 19% 2.6% 4.9% 18% 11% Page 12 of 19 5 3M ENVIRONMENTAL LABORATORY PROJECT NO.EOS-0442 Table 7. Matrix Spike Recoveries (continued and corrected for endogenous target analytes) A n alyte TDHA S a m p le ID LS 0505115-1 LS_0505115-2 LS 0505115-3 HS 0505115-1 HS 0505115-2 HS 0505115-3 Nominal Cone., ng/mL 0.302 0.302 0.302 3.02 3.02 3.02 O bserved C o n e ., ng/mL Endogenous Level in Matrix, ng/mL O bserved C o n e ., Corrected for Endogenous Level, ng/mL Recovery, % 0.392 0.354 0.334 2.93 2.84 -. 2.74 0.00641 0.00641 0.00641 0.00641 0.00641 0.00641 0.39 128% 0.35 115% 0.33 108% 2.9 97% 2.8 94% 2.7 91% % RSD 8% 3% C11 ACID LS 0505115-1 LS 0505115-2 LS 0505115-3 HS 0505115-1 HS 0505115-2 HS 0505115-3 0.297 0.297 0.297 2.97 2.97 2.07 0.355 0.292 0.310 2.89 2.82 2.69 0.0168 0.0166 0.0166 0.0168 0.0168 0.0168 0.338 0.275 0.293 ' 2.87 :* 2.80 I 2.67 114% 93% 99% 97% 94% 90% 11% 3.6% RFBS LS 0505115-1 LS 0505115-2 LS 0505115-3 HS 0505115-1 HS 0505115-2 HS 0505115-3 0.304 0.304 0.304 3.04 3.04 3.04 10.8 0.314 0.342 2.97 3.10 3.14 0.00815 0.00815 0.00815 0.00815 0.00815 0.00815 10.8' 0.306 0.334 2.96 3.09 3.13 3550%* 101% 110% 97% 102% 103% 6.2% 2.9% THPFOS LS 0505115-1 LS 0505115-2 LS 0505115-3 HS 0505115-1 HS 0505115-2 HS 0505115-3 0.312 0.312 0.312 3.12 3.12 3.12 NR NR 0.589 2.94 3.06 2.44 | 0.0256 0.0256 0.0256 0.0256 0.0256 0.0256 NR NR NR NR 0.56 181% 2.9 93% 3.0 ! 97% 2.4 77% NR 12% THPFDS LS 0505115-1 LS 0505115-2 LS 0505115-3 HS .0505115-1 HS 0505115-2 HS 0505116-3 0.302 0.302 0.302 3.02 3.02 3.02 0.330 0.306 0.302 2.94 2.85 2.94 0.0138 0.0138 0.0138 0.0138 0.0138 0.0138 0.316 0.292 0.288 2.93 2.84 2.93 105% ! 97% ! 95% 97% 94% 97% 5.1% 1.8% This value can be eliminated as an outlier at the 99% confidence level and will be excluded in all further calculations. N R = N ot Reported because measurable amounts o f target analyte were not observed. Page 13 o f 19 3M ENVIRONMENTAL LABORATORY PROJECT N0.E05-Q442 4 .5 Accuracy The accuracy for each analyte reported was calculated by taking the average o f the matrix spike recoveries for that analyte. The results are presented in Table 8. Table 8. Method Accuracy for Individual Analytes Analyte NFPA PFHA TDHA PFOA C9 Acid C10 Acid C11 Acid C12 Acid % Accuracy 21% 17% 5% 27% B% 1% 2% 14% Analyte FBSA FOSA PFBS PFHS PFOS THPFOS THPFDS % Accuracy 8% 2% 3% 4% 21% 12% + 3% 4 .6 Sam ple R elated Com m ents The extraction and analysis o f the samples was performed utilizing a non-validated method. The matrix spike results, PFOA [l,2-l3C] and PFOS [l8Oi] surrogate recoveries, and the calibration curve should be considered when evaluating the accuracy and precision of the sample results. 5 Individual Sample Data Tables 9 and 10 summarize individual sample data. Table 9. Perfluorinated Acid Results Sample ID NFPA (ng/mL) PFHA (ng/mL) TDHA (ng/mL) PFOA C9 Acid C10 ACID C11 ACID (ng/mL) (ng/mL) (ng/mL) (ng/mL) C12 ACID (ng/mL) LLOQ (ng/mL) 0.102 0.00505 0.00504 0.0991 0.0150 0.0259 0.00992 0.00997 TNA 6840 0.445 < LLOQ < LLOQ 3.89 0.656 0.198 TNA6843 1.51 < LLOQ 0.0752 0.509 0.127 0.0768 TNA 6847 1.56 0.0133 0.118 3.55 0.756 0.309 TNA 6934 0.866 0.00633 0.0232 5.72 1.19 0.450 TNA 6935 1.04 0.00838 0.0964 3.48 1.17 0.551 TNA 6944 1.43 < LLOQ 0.290 5.67 1.18 0.584 TNA 6959 0.795 0.00964 0.195 2.52 1.96 0.193 TNA 6960 0.932 0.00869 0.210 2.91 0.611 0.300 TNA 6967 0.450 < LLOQ 0.270 3.66 0.531 0.209 TNA 6973 0.680 0,00652 0.352 3.60 0.940 0.391 TNA 6975 0.682 0.00779 0.398 4.02 0.906 0.402 The uncertainty o f these results (based on isotopically labeled surrogate recovery) is +/- 34%. 0.0968 0.0259 0.141 0.188 0.384 0.196 0.734 0.121 0.109 0.124 0.149 0.0282 0.0212 0.0207 0.0264 0.0833 0.0465 0.0370 0.0190 0.0198 0.0328 0.0295 Page 14 of 19 3M ENVIRONMENTAL LABORATORY PROJECT NO.EOS-0442 Table 9. Perfluorinated Acid Results (continued) Sample ID NFPA iM 'm jy PFHA TDHA PFOA C9 Acid (ng/mL) (ng/mL) (ng/mL) (ng/mL) C10CID (ng/mL) C11 ACID (ng/mL) C12ACID (ng/mL) LLOQ (ng/mL) 0.102 0.0050S 0.00504 0.0991 0J31S0 0.0259 0.00992 0.00997 TNA6977* TNA 6981 1-23 0.327 < LLOQ 0.0442 LLOQ 0.0555 0.406 2.26 0.0950 0.423 0.0374 0.147 0.0152 0.0708 < LLOQ < LLOQ TNA 6992 1.09 < LLOQ 0.239 3.32 0.842 0.319 0.158 0.0124 TNA 6994 1.75 0.00694 0.0750 0.770 0.206 0.0830 0.142 0.0223 TNA 6995 1.06 <LLOQ 0,198 2.00 1.41 0.169 0.688 0.0147 TNA 6997 1.32 < LLOQ 0.179 2.33 1.97 0.202 0.637 0.0185 TNA 7000 1.08 < LLOQ 0.174 3.92 0.717 0.332 0.112 0.0240 TNA 7006 0.818 < LLOQ 0.166 3.32 0.686 TNA 6938 1.09 < LLOQ 0.474 4.36 1.20 0.306 0/421 0.0946 0.165 0.0128 0.0150 TNA 6939 TNA 6941 TNA 6950 TNA 6953 0.615 1.02 2.11 1.27 0.0323 < LLOQ < LLOQ < LLOQ 0.0826 0.0608 0.0925 0.0442 2.01 4.07 3.20 3.08 1.09 1.61 0.792 0.785 0.344 0.599 0.22S 0.249 0.28 0.424 0.116 0.0753 0.0302 0.0527 0.0115 0.0150 TNA 6955 TNA 6962 TNA 6964 TNA 6966 1.26 1.09 1.28 0.965 < LLOQ < LLOQ < LLOQ < LLOQ 0.111 < LLOQ 0.259 < LLOQ 2.20 3.51 4.00 2.81 0.603 0.690 0.933 0.562 : 0.341 0.311 0.386 0.249 0.196 0,111 0.117 0.0916 0.0309 0.0183 0.0176 0.0195 TNA 6B70 0.781 <LLOQ 0.159 4.17 0.917 0.322 0,148 0.0119 TNA 6972 TNA 6978* TNA 6979 TNA 6982 TNA 6984 1.19 1.36 1.11 0.751 0.899 <LLOQ < LLOQ <LLOQ <LLOQ < LLOQ 0.123 0.0481 0.126 0.0660 0.0636 3.79 0.359 3.21 2.16 2.19 0.761 0.114 0.560 0.653 0.556 0.257 0.0274 0.304 0.173 0.261 0.126 0.0152 0.156 0.0547 0.184 0.0192 < LLOQ 0.0161 < LLOQ 0.0108 TNA 6986 0.500 <LLOQ 0.0549 2.31 0:460 0.179 0.0394 LLOQ TNA 6988 0.916 < LLOQ 0.0443 1.93 0.610 0.239 0.164 tLLOQ Mean M adian High Low 1.04 1.05 2.11 0.327 0.0111 0.00838 0.0323 0.00613 D.151 0-118 0.474 0.0232 2.98 3.21 5.72 0.359 0.813 0-737 1.97 0,095 0.282 0.281 0.599 0.0274 0185 0.134 0.734 0.0152 0.0246 0.0197 0.0833 0.0108 Standard ' D eviano 0.378 0.00822 0.111 1.27 0,445 0.137 0.176 0.0149 S O F H IT S 36 9 33 36 36 36 36 30 TN-A-6977 and -6978 ate from the same lot of pooled serum. The uncertainty of these results (based on isotopically labeled surrogate recovery) is +/- 34%. Page 15 o f 19 3M ENVIRONMENTAL LABORATORY PROJECT NO.E05-0442 Table 9. Sample Results for Additional Perfluorfndted and Highly Fluorinated Analytes Sample ID FBSA FOSA PFBS PFHS PFOS THPFOS THPFDS (ng/mL) (ng/mL) (ng/mL) (ng/mL) (ng/mL) (ng/mL) (ng/mL) LLOQ (ng/mL) 0.00501 0.00498 0.00506 0.0158 0.103 0.00520 0.00503 TNA 6840 < LLOQ 0.356 0.00795 3.07 21.9 TNA 6843 < LLOQ 0.124 0.00854 0.606 2.78 TNA 6847 < LLOQ 0.0694 0.00854 1.62 22.4 TNA 6934 < LLOQ 0.314 0.0112 1.95 27.0 TNA 6935 < LLOQ 0.133 0.0101 1.75 14.5 TNA 6944 < LLOQ 0.0786 0.00969 3.66 16.9 TNA 6959 < LLOQ 0.232 0.00979 2.85 12.4 TNA 6960 < LLOQ 0.166 0.0107 3.27 14.6 TNA 6967 0.00542 0.446 0.0238 3.04 19.7 TNA 6973 <LL0Q 0.156 0.0172 3.94 16.9 TNA 6975 < LLOQ 0.127 0.00858 3.97 18.0 TNA 6977* < LLOQ 0.118 0.0840 0.662 2.03 TNA 6981 < LLOQ ' 0.122 < LLOQ .1.17 12.5 TNA 6992 TNA 6994 < LLOQ 0.159 0.0103 3.66 16.6 < LLOQ 0^116 0.0211 1.37 2.78 TNA 6995 < LLOQ 0.248 0.0160 2.88 11.6 TNA 6997 < LLOQ 0.224 0.00942 2.92 12,0 TNA 7000 < LLOQ 0.0906 0.0148 3.97 17.5 TNA 7006 < LLOQ 0.202 0.00989 3.43 19.2 TNA 6938 < LLOQ 0.0446 0.00711 2.38 16.3 TNA 6939 < LLOQ 0.0967 0.0202 1.65 6.43 TNA 6941 < LLOQ 0.130 0.00991 2.90 21.8 TNA 6950 < LLOQ 0.0378 0.00545 2.84 14.9 TNA 6953 < LLOQ 0.0459 0.00804 4.08 21.9 TNA 6955 < LLOQ 0.0961 0.00588 2.73 9.82 TNA 6962 < LLOQ 0.162 0.0122 3.64 17.8 TNA 6964 TNA 6968 < LLOQ < LLOQ 0.187 0.135 0.0218 0.00802 4.18 3.10 20.1 15.2 TNA 6970 < LLOQ 0.238 0.00737 3.22 18.3 TNA 6972 < LLOQ 0.124 0.0557 2.82 14.4 TNA 6978* < LLOQ 0.109 0.00577 0.640 2.21 TNA 6979 < LLOQ 0.121 0.0127 1.97 19.0 TNA 6982 < LLOQ 0.0903 0.0144 1.53 14.7 TNA 6984 < LLOQ 0.108 < LLOQ 1.32 14.2 TNA 6986 < LLOQ 0.0712 0.0567 1.57 14.1 The uncertainty o f these results (based on isotopically labeled surrogate recovery) is +/- 34%. < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ <LLOQ < LLOQ < LLOQ < LLOQ < LLOQ cLLOQ <LLOQ < LLOQ < LLOQ < LLOQ < LLOQ <LLOQ <LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ <LLOQ <LLOQ < LLOQ < LLOQ < LLOQ <LLOQ <LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 0.0562 0.106 0.126 0.00772 . 0.0457 0.048 0.0311 0.0365 0.0425 0.0353 0.0366 0.0413 0.0351 0.0417 0.0249 0.028 0.0273 0.034 0.0358 0.0259 0.0349 0.0279 0.0753 0.0273 0.0287 0.0345 0.0461 0.0281 0.0466 0.0319 0.0485 0.0799 0.0581 0.0529 0.0459 Page 16 of 19 3M ENVIRONMENTAL LABORATORY PROJECT NO.EOS-0442 Table 9. Sample Results for Additional Perfluorinated and Highly Fluorinated Analytes (continued) Sample ID FBSA FOSA PFBS PFHS PFOS THPFOS THPFDS (ng/mL) (ng/mL) (ng/mL) (ng/mL) (ng/mL) (ng/mL) (ng/mL) LLQQ (ng/mL) 0.00501 0.00498 0.00506 0.0158 0.103 0.00520 0.00503 TNA 6988 Mean M edian High Low < LLOO NA NA 0.00542 0.00542 0.101 0.149 0.124 0.446 0.0378 < LLOQ 0.01645 0.0101 0.084 0.0055 1.45 2.55 2.85 4.18 0.606 15.7 < LLOQ 0.0586 14.9 NA 0.0442 15.5 NA 0.0366 27 NA 0.126 2.03 NA 0.00772 Standard Deviation NA 0.0876 0.0169 1.07 5.94 # O F HITS 1 36 33 36 36 TN-A-6977 and -6978 are from the same lot o f pooled serum The uncertainty o f these results (based on isotopically labeled surrogate recovery) is +/- 34%. NA 0.0227 0 36 Table 10. Isomer Ratios for Individual Samples Sample ID TNA 6840 TNA 6843 TNA 6847 TNA 6934 TNA 6935 TNA 6944 TNA 6959 TNA 6960 TNA 6967 TNA 6973 TNA 6975 TNA 6 9 7 7 " TNA 6981 TNA 6992 TNA 6994 TNA 6995 TNA 6997 TNA 7000 TNA 7006 TNA 6936 TNA 6939 TNA 6941 TNA 6950 TNA 6953 Sample Origin California Missouri Pennsylvania Pennsylvania New York Michigan Virginia Virginia Maryland Montreal. Q uebec Montreal, Quebec M assachusetts Texas California California California California California California New York New York New York Michigan Michigan PFOA % Branched isomer <LLOQ* LLOQ* 3.2% 1.3% 6.8% 2.7% 3.9% 2.3% LLOQ* 2.3% 2.3% <LLOQ* LLOQ* LLOQ* 3.1% 4.2% 3.9% 2.2% LLOQ* 2.3% 1.1% 0.7% 1.6% 0.8% PFOS % Branched Isomer 37% 35% 42% 46% 34% 33% 37% 36% 40% 39% 40% 35% 40% 36% 38% 39% 39% 40% 39% 40% 33% 35% 37% 39% Page 17 of 19 3M ENVIRONMENTAL LABORATORY PROJECT NO.E05-044Z TNA 6955 TNA 6962 TNA 6964 TNA 6906 TNA 8970 TNA 6972 TNA 6978" TNA 6979 TNA 6982 TNA 6984 TNA 6986 TNA 69B8 Michigan Virginia Virginia Virginia Maryland Maryland M assachusetts M assachusetts Texas Texas Texas Texas 16% <LLOQ* LLOQ* LLOQ* <LLQQ* 2.1% LLOQ* 3.5% 0.8% <LLOQ* <LLOQ* LLOQ* 34% 39% 39% 37% 38% 36% 36% 30% 41% 30% 41% 33% Average: 2.5%*** 37% Standard Deviation 1.4% *" 3.3% *The branched isomer in the sample was < LLOQ **TN-A-6977 and -6 9 7 8 are from the same lot o f pooled serum. ***The average and standard deviation represent only those samples with isomer values above the LLOQ. 6 Data/Sample Retention All original raw data and the analytical report have been archived at the 3M Environmental Laboratory. The test materials and analytical reference standard reserve samples, as well as the samples pertaining to this project are archived at the 3M Environmental Laboratory. Reserve samples, digital copies o f original data and all original paper data will be retained in the archives of 3M Environmental Laboratory for a period o f at least 10 years following report signing. 7 Conclusion Under the conditions o f the study, endogenous fluorochemicals were found to be present in all samples. NFPA, PFOA, Cc>Acid, Cio Acid, Cu Acid, FOSA, PFHS, PFOS and THPFDS were detected in every lot of serum. TDHA and PFBS were detected in all but three samples, and the C u Acid was detected in 30 of 36 samples. Conversely, PFHA was only detected in 9 o f 36 samples, and FBSA was only detected in 1 of 36 samples. THPFOS was not detected in any o f the samples using the present methodology. PFOA [l,2-n C] surrogate recoveries were all within 30% while PFOS [1802] surrogate recoveries were all within +/- 34%. Method accuracy for individual analytes ranged from +/-1% to +/- 27% while overall method uncertainty (based on isotopically labeled surrogate recovery) was +/- 34%. Page 18 of 19 3M ENVIRONMENTAL LABORATORY PROJECT NO.E05-0442 The 3M Environmental Labocatoiy's Quality Assurance Unit has audited the data and report for this project n A SL Quality Assonance Representative la-i-aoos D ate Page 19 of 19