Katherine E. Reed, 3M Environmental, Health and 900 Bush Avenue, Building -. ,7 Staff Vice President Safety OperatiOns PO Box 33331 St. Paul. MN 55133-3331 6517784331 7:5 HQ, otosaeo 394,1 August 26, 2005 NO TSCA CBI Document Processing Center (Mail Code 7407M) Room 6428 . Attention: TSCA 8(e) Coordinator - Of?ce of Pollution Prevention and Toxics- - a h. 39 US. Environmental Protection Agency v. 1201 Constitution-Avenue, N. W. anai? Washington, D. C. 20460 NO Re: TSCA Section 8(e) Supplemental Notice: Sulfonate- asedand Carboxylic-based luorochemicals, Docket Nos. SEH Dear Sir: 3M is submitting this notice to supplement its prev1ous submissions on sulfonyl and carboxylic?basedp fluorochernicals. . 3M has recently received the enclosed analytical report Which presents data from 36 lots of commercially available normal pooled human serum that were purchased by the 3M Environmental Laboratory and subsequently screened for endogenous ?uorochemicals. Each sample was analyzed using a protein precipitation technique and screened for sixteen different ?uorochemicals, including per?uorooctane sulfonate (PFOS) (CanSOg') and per?uorOoctanoic acid (PFOA) (C717 15COOH), using liquid chromatography/tandem mass spectrometry Much of the data presented in the report was derived from extrapolated curves and should be considered as screening quality data only and not necessarily representative of the general population levels. Not all of the detected chemicals are or were produced by 3M. In addition, the levels of ?uorochemicals reported are generally corroborative of previously reported values and/or do not represent ?substantial risk? information If you have any questions, please do not hesitate to contact Dale L. Bacon at (651). 778-4736. Sincerely, Wt?ef Katherine D. Reed, Staff Vice President Environmental Technology and Safety Services 8 BEST COPY AVAILABLE Hm, 3M ENVIRONMENTAL LABORATORY PROJECT NO. EDS-0120 3M - NO TSCA CBI Analytical Report Revised August .,12 2005 Initial Quantitative Screening of Commercial LOts of Human Serum Obtained In 2004 for Endogenous Fluorochemlcals Using Protein Precipitation and Liquid Chromatography/Tandem Mass Spectrometry Laboratory Request Number: ?Testing Laboratory 3M Environmental Health and Safety Operations 3M Environmental Laboratory 3M Environmentai Laboratory Project Lead Mark E. Ell?fSon? "Btiilding 245509; PO Box 33-31"- Saint Pant, MN 551 315-3331 Phone: 651-778-5405 I Testing Facility Management I William K. Reagan Ph. 0. . Budding PO Box 3331 Saint Paul MN 55133-3331 Phone: 651-778-6565 Requester Dale Bacon Building 2-3E-09 PO Box 3331 Saint Paul, MN 55133-3331 Phone: 651 ?778-4736 Page 1 of 18 3M ENVIRONMENTAL LABORATORY PROJECT 20 3M Environmental Laboratory Analytical Report Project Number: HOS-0120 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 ?uorochemicals. Thirty?six separate lots of pooled normal human serum were extracted using a protein precipitation technique and screened for sixteen di?'erent ?uorochemicals using liquid chromatography/tandem mass spectrometry Serum samples were received from various suppliers between 9/9/04 and 12/2/04. Samples were analyzed between 02/03/05 and 02/09/03 . This range??nding study was designed to provide lower limits of quantitation (LLOQs) of approximately 100- 300 ppt. Upon review of the data, it was discovered that many of the target analytes were present at Concentrations much lower than the 100 300 target level. As an interim solution and until a mere de?nitive study can be completed, signal to noise ratios were evaluated and a limit of detection (LOD) was de?ned for each analyte. As a result, calibration curves were extrapolated between the LLOQ and the newly de?ned LOD, allowing screening A quality data to be determined for some analytes as low as 19. 9 ppt. Therefore, it must be emphasized that much of the dam presented in this repert was derived from extrapolated curves and should be considered as screening quality data only. Sample results are summarised in Table l. The average values reported represent samples in which there was detectable analyte. Sample values that were less than the limit of detection were not included in the average value calculations. Please refer to Section 5 for individual sample results. Page 2 of 18 Table 1. Summary of Per?uorinated Acid Results 3M ENVIRONMENT AL LABORATORY PROJECT Mean' 3.3-3 2.345 4.6.17 1.25" NR2 0.623 Median 3.33 2.6} NR2 0.954 4.59 1.09 . NR2 0.660 .ngh 7.06 3.56.7 NR2 fill-.02 .1260? 2.6.3 0.642 Low 0.507 2.45" 0.680 0.477 0.337 Na? 0.312 Standard Deviation 2.17 N33. 0.0633- 3.64. 0.571: NR2 0.206 Number of Samples with_ .. - I 3? . 1. Madam Mam 2-0136 3:66 24.66 34136 0/38 I-II 6/36 0/36 Not Detected. This analyte was not detected at any of the samples. Not Reported This analyte was- idetE-cf'ed in one ofthe 35 mics but. appears to be a contaminant or sampl'c?will mam mad the resultswill hereported an the de?nitive?sturdy: Table 2. Summary of Other Pertluonnated and l-liIghlyI Fluorlnatod Analyto Results :r0.0639 . 40.0819 woof-0.1 2.71 - 97% 0.998 5/ Page 4 of 13 3M ENVIRONMENTAL LABORATORY PROJECT MOI-3050120 The samples for this study were prepared as l. The serum samples were allowed to thaw and warm to room temperature. 2. One milliliter of serum (or water in the case-- of method blanks) was 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 of Interest. In ?30 11110.1 was added as a .7 sun'cgate to all serum samples calibration standards matrix blanks, and method blanks at a Concentration of 3 .01 11g/mL to evaluate extraction method performance . .. - . . 4. 1 Two milliliters of 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 centri?iged at approxunately 3000 ref for 20 minutes to pellet . the precipitate . .6. A?er removal item the centrifuge, the supernatant was transferred to an tautovial for . Quanti?cation Was by high performance liquid chromatography tandem mass spectrometry The. fellow'ing parameters Were used.- Liquid Chromatograph Hewlett~Packard? Series 1100 Liquid Chromatograph system 3 Analytical Column: Cu; 2x100mm, Sum particle size Column temperature 30 Stop Time; 20 0 minutes Flow rate: 300 1111111111 Injection'VOlume: 5 Mobile phase. components Solvent A: 2 .0 111M ammonium acetate in ASTM Type I water Solvent B: HPLC Grade Methanol . 3 . . Solvent Gradient 0.00 20% 1.110 20% 14.50 90% 7.15.511 90% 16.50 . 20%- 20.00 20%" Page 5 of 18 3M ENVIRONMENTAL LABORATORY PROJECT NO. 505?0120 Mass Spectrometer: MDX Sciex? API 4000Q-Trap Mass Spectrometer Detector So?tware: Version 1.4 Ionization Mode: Electrospray Negative Temperature: 450 GS 35 G82: 45 Entrance Potential: 10 Analysis Type: Multiple Reaction Monitoring (MRM) Table 5. Acquisition Parameters 168.9 I 262.9 . -3o .PFHA(Caacid) . 313.0 2681,1189 . .- . 45 TDHA (c:7 Add) 362.9. . 3181,1688, 118.9 -35 413.0 ~40 CgAcid 463.0 . II 416.7, 268.9, 218.9 -50 CwAcid 1 612.9 I 269.1 - . -100. 0,1 Acid. 563.0 518.7, 268.9, 218.8 . 45 CuAcid- . 613.0 I -50 FBSA 297.9 - I 78.0 i -55 FOSA 497.9 . 77.9 4" -90 PFBS 298.9 989.799 455 . PFHS 398.9 98.9, 80.0 -95 PFOS 498.9 80.0, 992, 130.0 -60 THPFOS 426.9 406.8. 80.9 -70 I I 526.9 . 506.7, 81.0. I :10 PFOA [1.2130] 414.9 . 369.8 . .- +10 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. .1 - . 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 sar'ne manner as the samples. Stock solutions were individually prepared from neat ?uorochemicals and diluted in methanol (except PFBA, which was prepared in Milli-Q water to prevent esteri?cation from occurring). Samples were quanti?ed versus -an 8-point extracted calibration curve. The regression analysis was performed using a quadratic curve ?t weighted l/x. Page 6 of 18 3M ENVIRONMENTAL LABORATORY PROJECT NO. 505-0120 Calibration standards were Within 20% (i25% for the Lower Limit of Quantitation with a coet?cient of determination 995. IoW?level calibration standards, Which were not Within i25%, were net. included 111 the calibration curve. Calibration standards that were not greater than twice the methOd blanks Were not included 11: the calibration curve. 4.1.2 Continuing Calibration Veri?cation (CCV) A Continuing Calibration Veri?cation (CCV) was run at least once every 10 samples, bracketing the samples With the calibration curve The CCV recovery Was within for all analytes. 4.1.3 System Suitability Five system suitability standards were analyzed before the initial eahbration curve and after the last CCV. These samples had area counts with a of<5% and a retention time of When evaluated independently except for the following instancesz? . Several individual for the Cm Acid failed the retention-time system suitability criteria. However. the data reported was derived from the quantitation of. the TIC, which passed system suitability requirements. All of the system suitability sets for FBSA area counts failed The were 6 7. 5% and 8. However, no samples had area counts above the LLOQ for this analyte therefore the data Was accepted. 4..14 LowerLumtonuantrtahonaLOQ) . .. . The LLOQ for each analyte Was equal to the lowest standard in the calibratlon curve 125% - . 7 . . 4..15 LnnitofDetectIonaBD) The LOD for each analyte was equal to the area counts (converted to a concentration) that: were 2?3 times above the method blank. 4.1. 6 Blanks . Method Blanks All ofthe method blanks 1.11 this studyhad area counts 50% 'ofthe LLOQ. Matrix Blanks A - Two matrix blanks were prepared 1n rabbit serum and analyzed with the samples. One matrix blank contained endogenous levels of PFOS that. were above the LLOQ. An average endogenous value for PFOS was determined by substituting a number equal to 1/2 the LLOQ for 003 57 Page 7 :1le 3M ENVIRONMENTAL LABORATORY PROJECT 510505-0120 Table 6. Average Endogenous PFOS Value Determination LLoo?ngL) 4 . (Laos Blk050203?003 LL00 Blk050203?004 . A . 0.477 Average: 0.315 The arerage endogenous PFOS value was used to correct the matrix spike response for endogenous levels of PFOS in rabbit semrn. 4.1.7 Solventhxanks . a . . Aoetoniu'ile blanks were analyzed throughout the analytical 'Analyte response was les?sthanl/ztheLLOQina?cases? . '7 W7 7 Surrogate . 13C PFOA was. added as a surrogate to "all serumsamt?estcalibration standards, matrix blanks, and method blanks at a concentratlonof to evaluate the extraction method performance. All. surrogate recoveries were within t20%. 4.2 Laboratory Control Spikes Six Laboratory Control Sriikes(LCS) were prepared inrabbit serum and- analyzed with the samples.- Results were corrected. forendogenous: loitels of analyte in the matrix (Where appropriate), and are presented below. Table 7- M??trix Spike Recoveries in "112be Serum PFBA (c4 acid) . Loo - 0.426 LLoo - 3.00 mm NFPA (65- acid) LOB I 4.53 :1me LLOQ 15.3 NA Not Applicable Page 8 of 18 3M ENVIRONMENTAL LABORATORY PROJECT Masses-0120 Table 7. Matrix spam Recoveries in Rabbit Serum (continued), (cs Qc050203-003 LLOQ 15.1 00050203004 00050203005 96050203006 10ch: cosmos-003 LL00 a: 3.03 00050203404 - ?00050203005 mosthaoda ?03050203401 (20050203402 C10 Acm. LLoosanz-nymL ocosozos-omv- C1 1min. LLOQ 0.2597 {:me .0 Lunch a 0.147 NA wNot Applicablo- Page 9 of18 I0 3M ENVIRONMENTAL LABORATORY PROJECT NO. 5054}? 20 Table 7. Matrix Spike Recoveries in Rabbit Serum (continued) 00050203001 . 00050203002 "c PFOA 06050203003- 00050203004 00050203005 . . 00050203000 00050203001 1 060502024302 1 00050202003 raglle 00050202004 00050202005 CQAcid 00050202002 00050202003 mania-0293 mm. 00050202004 - 00050202405- - 00050202000; PFOA . LLOQ 0.303 FOSA LL00 0.300 Page 10 of 18 Table 7. Matrix Spike! Recoveries in Rabbit Serum (continua) PFBS LLOQ 0.303 was}. PFHs LLDQ a 0.315 nalmL LLOQ 0.780 mm! LLOQ .- 3M ENVIRONMENTAL LABORATORY PROJECT 20 Pagc110f18 3M ENVIRONMENTAL LABORATORY PROJECT NO.EOS-O12O Table 8. PFOS Matrix Spike Recovery in Rabbit Serum, Corrected for Endogenous Levels . a ocosczoz-om . 3.17 . ,aae- - . ecosozoz-aoz . 3.03 . . 2.721 . 3.1 - - 2479;. 0605020sz . a 177.4 . . 77.1- ocosozcz-oes . . ma - . "77.3 . . .. 76.1 75.8' The accuracy for-each anal?e reported was calculated bytalcin?g the average of the matrix spike recoveries for that ?analyte. The results are presented below. 4.3 Accuracy Table 9. Method Accuracy for Individual Analytes m. ., r. . we FOSA j. 7. we omens . aces PFBS 1m PFOA we. . PFHs .1254. 09 Acid . em PFOS was" 010 Acid" THPFOS 12% C11Acid ms. 11% "Ihc% ontheba'gh concentration sp?cerccovericswereless 4.4 Sample Related Comments The extraction and analysis of the samples was performed utilizing a non-Validated method. The matrix spike results, PFOA surrogate recoveries, and the extrapolated calibration curve (between the LLOQ and LCD) should be considered when evaluating the accuracy and precision of the sample results. Tables 10 and 11 summarize individual sample data Page 12 of 18 ll 3M ENVIRONMENTAL LABORATORY PROJECT NO. 505-01 20 Table 10. Per?uorinated Acid Results Page 13 of18 ML 3M ENVIRONMENTAL LABORATORY Table 10. Per?uorinated Acid Results (00ntinued) TNA6961 6.76 2.45 4.00 4.12 . 1.28 . 4.00 TNA6992 0.690' . 4.76.? 0.912 000 TNA6994 4.00