Katherine E. Reed, PILD. 3M Environmental, Health and Staff Vice President Safety Operations a. 1 v" wt?" W. e" Edi Ste? i ti}, .MzzE: Certi?ed Mail August 26, 2005 7:6 7y Document Processing Center (Mail Code 7407M) Room 6428 Attention: TSCA Of?ce of Pollution Prevention and Toxics US. Environmental Protection Agency 1201 Constitution Avenue, N. W. Washington, D. C. 20460 900 Bush Avenue, Building 42-2E-26 PO Box 33331 'St. Paul, MN 55133-3331 651 778 4331 4N0 TSCA 450 3 7% Dental/ii NO Re: TSCA Section 8(e) Supplemental Notice: Sulfonate?based and Carboxylic-based luorochemicals, Docket Nos] 73 Dear Sir: 3M is submitting this notice to supplement its previous submissions on sulfonyl and carboxylic-based ?uorochernicals. 3M has recently received the enclosed analytical'report whichpresents data from 36 lots of commercially available normal pooled human serum that werei'purchased by the 3M Environmental Laboratory and subsequently screened for endogenous fluorochemicals. EaCh sample was analyzed using a protein precipitation technique and screened for sixteen different ?uorochemicals, including per?uorooctane sulfonate (PF OS) 17803') and per?uorooctanoic acid (PFOA) 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. Nor 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 anyquestions, please do not hesitate to contact Dale L. Bacon at (651) 778-473 6. Sincerely, 'm a Katherine D. Reed, 4g;- Wm 5? minimmillin-gun-yinmumnum Environmental Technology and Safety Services BEST COPY AVAILABLE at; :n 3M ENVIRONMENTAL LABORATORY PROJECT moaoemzo 3M - INOTSCACBI Analytical Report August 12 2005 Initial Quantitative Screening of Commercial Lots of Human Serum Obtained' In 2004 for Endogenous Fluorochemicals USing Protein Precipitation and Liquid Chromatography/Tandem Masa Spectrometry LaboIatory ReqUeStNumberz' EDS-0120 I . - Laboratdry 3M Enwronmental and Safety Operations 3M EnvironmentaI Laboratory I 1 3M .EnViI'onmenIaI tang-awry Lead Mark Ellefson 09 PO Box 3331 - Saint Paul; MN 55133-3331 Phone 651-778 5405 Testmg Facility Management 1 William Reagan Ph D. I Budding 09 4 PO Box 3331 Semi Paul MN 55133?3331 Phone. 651?778?6565 Requester Dale Bacon Building - PO Box 3331? Saint Paul, MN 55133-3331 Phone: 651-778-4736 Page 1 of 18 3M ENVIRONMENTAL LABORATORY PROJECT NO. EDS-01 20 3M Environmental Laboratory Analytical Report Project Number: 1505-0120 The following analytical report presents data Iavailablenornial pooled human serum that were purchased by the 3M Environmental Laboratory and subsequently screened for endogenous ?uorochemlcals Thirty- -six separate lots of pooled normal human Serum Were extracted using Ia. protein precipitation technique and screened for sixteen different ?uorochernicals using liquid chromatography/tandem inass spectrometry Serum samples were received from various sItippliers between 9/ 9/04 and 12/2/04 Samples were analyzed between 02/03/05 and 02/09/05 - This range-?nding study was designed to provide lower lirnits of quantitation (LLOQs) of approximately 100- 309 ppt. Upon review of the data, it was discovered that many of the target analytes were present at concentratlons lower than the 100- 300 target level. As an interim sole/non and tintil a more de?nitive stInIdy I can be completed signal to noise ranos were evaluated and a limit of detection I I (LOID) was de?ned for each analyte a Ire-stilt, callbranon curves were a I . extrapolated between the LLOQI and, the newly de?ned LOD al1owing screening I quality data to be determined for some Ianalytes as low as 19. 9 Therefore, it ?j must be emphasized that ranch of the data presented 111 this repbrt was deriVed ?orn extrapolated curves and should be censidered as screening quality data only. Sample results are summanzed 111 Table The average valnes reported represent samples In Which there was detectable analyte Sample values that were less than the limit of detection were not mcluded 1n the average valneI calculanons Please refer to Section 5 for individual sample results Page 2 of 18 3M ENVIRONMENTAL LABORATORY PROJECT {00505-0120 Table 1. Summary of Perfluorinated Acid Results . Detectable Analyte . . 2 20136 0/46?! 1712s]- "21122 I0623 ?stem Median 6.33 2.5-. ?0.954 459 1-1697 NR3 [0 660 999!n 0.998 Page 4 of E8 3M ENVIRONMENTAL LABORATORY PROJECT The samples for this study were prepared as follows: 2. The serum samples were allowed to thaw and warm to room temperature One milliliter of serum (01 water in the case of method blanks) was added to a - 15 mL serew-capped polypropylene centrifuge tube. Matrix spikes and method blanks were prepared by spiking the serum (or ?water) With the analytes of mterest In addition, 3 PFOA was added as a i' JJsurrogate to all serum samples calibration standards matrix blanks, and JthJe precipitate methOd blanks at a concentration of 3. 01 ng/mL to evaluate extraCtiOn method performance -. . . . milliliters 'of acetomtrile Were added to the serum and the centrifuge tubes were capped and vortex mixed, allowing the serum proteins to precipitate. The tubes Were centrifugedat apl'J? __oxrmately 3000 1111' for 20 minutes to pellet After removal ?omithe centri?rge the supernatant was transferred to an ,autovral for analysm 1 Quanti?cation Was accomplished by high performance liquid chromatography tandem mass spectrometry The. followmg parameters were need: - Liquid Chromatograplr Hewlett?Packard? Series 1100 Liquid Chromatograph . ?system Analytical column Keystone C13 2x100mm, 511m particle size JC011111111 temperature 30 3C 5 - . . Stop Time; 20. 0 minutes 'Flowrate 30.0 uL/mm Injection volume 5 11L Mobile phase components -- Solvent A: 2 0 mM ammonium acetate in ASTM Type I water g; . SolventB HP Grade Methanol -: . . -- Solvent Gradient I i 11-51% ~21! -2091?. 100 . 14.50_ 15so ?16150 20%: 20.00 . Page 5 of.18 3M ENVIRONMENTAL LABORATORY PROJECT 'Mass Spectrometer: MZDX Sciex? API 4000 Irap Mass Spectrometer Detector _-So?ware: Version .4 Ionization Mode: Electrospray Negative Temperature: 450 i -. 35 45 Entrance Peteutial: 10 Analysis Type: Multiple Reaction Monitoring (MRM) Table 5. Acquisitieh Parameters PFBA 213.9 . .. ACsAcid 292.9 219.9") ..PFHA 1 . 313.9: TDHA c7 . 392.9 . 319. 4999,1199 PFOA C9 . 1 413.9 . .9 219.9. 1999? CgAeid i . 1 493.9 .- 4 9.7 299.9 219.9 Acid .. 12.9 299.15 Acid. . 593.9 .5197 299.9 219.9 . Acid 913.9 7199 319.7? FBSA 297.9- . 79.9 FOSA 497.9 1 77.9 PFBS "299.9 . 99.9;799 PFHS . .- 7 "99.9, 99.9 I 499.9. 7992.11399; THPFOS . 426.9 4999.809 414.9 . 399.9 . When multiple transitions were monitored, the total ion current (TIC) was used'to quantify each analyte, thereby providing greater sensitivity than indiVidual Q3 1911 measurements. . . . 4.1 Calibration -. 4.1.1 Calibration Curve Ari extracted calibration curve was formed by spiking rabbit serum at prescribed levels and extracting the curve {in the same manneras the samples. Stock solutions were individually prepared ?om.neat ?ucrochemicals and diluted in methanol (except PFBA, which was prepared in Milli-Q water to prevent esteri?cation ?om occurring). Samples were quanti?ed versus an 8-point extracted calibration curve. The regression analysis was performed using a quadratic curve ?t weighted UK. Page 6 of 18 3M ENVIRONMENTAL LABORATORY PROJECT NO. 505?0120 . Calibration standards were within :t 20% (125% for. the Lower Limit of Quantitation With a coef?cient of determination 99-5. Low-leizel calibration standards, Which were not W1thin i25%, were not included in the calibration curve. Calibration standards that were not greater than twice the method blanks were not included 111 the calibration curve . . 4.1.2 Continuing Calibrahon 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 +25% for all analytes . 4.1.3 Systein Suitability Five system suitability standards Were analyzed before the initial calibration 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 IrolloWing instancesSeveral individual transitioas monitored for the C10 Ac1d 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 FIBSA area counts failed The were 7. 5% and 8 However, no samples had area counts above the LLOQ for this analyte therefore the data Was accepted. . 4.1.4 .LoWer Lanit of Qnantita?on (LIDQ) - t'Irhe LILOQ for each analyte Was equal to the loWestI standard In the calibration curve I - that met accuracy requirements of 75% 125% 4.1.5 Limit ofDetectlon (LOB) The LCD for each analyte Was equal to the area counts (converted to a concentration) thai Iwere 3 nines above the method Ibl'an'k. . - 4..1 I6 Blanks MethOd Blanks I . jg: All of the method blanks? in this study had area counts 50% of the Matrix Blanks '7 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 FF OS was determined by a number "equal to 1/2 the LLOQ for 003. - Page 7 oflS 3M ENVIRONMENTAL. LABORATORY PROJECT 0. 505-01 20 Table 6. Average Endogenous Determination . wooing-1mm. - . . . Blk050203?003 f- . <-L-Loo~ awesome?004- ., . 1 . 110.477 Average: I I 0.315 The average endogenous PFOS value was used to correct the matrix spike 7 response for endogenoos levels of FFQS 1n rabbit Sm. 4.1.7 Solvent Blanks Acetonitrile b1ank_s were analyzed throughout the analytical run. Analyte response was lesstl1an1/2theLLOQmallcases -r . 4.1.8 Sui-regatta 13C PFOA was added as a surrogate to all serum samples calibratioh standards, matrix blanks, and method blanks at a concentratmn of 3. 01 hg/mL to evaluate the extraotion method performance All surrogate reeoYeries Were Within i20% 4.2 Laboratory Control Spikes (LCS) Six Iabora?tOry control spikes (LCS) were prepared 111? rabbit serum and analyzed With the samples.- Results We1e caneeted for endogenous leYels of analme 111 the matrix (Where appropriate), and are presented below 1 Table,T-MMatrIerpnke Recoveries in' Rabbit Serum .I - PFBA (c4 acid) 1- . - .LOD II .426 11me LLOQ 3 .00 ngImL 900502034303 .. -- NFPA (CS acid) LOD 114,53 :1me 151.3 NA Not Applicable Page 8 of 18 3M ENVIRONMENTAL LAB ORATORY PROJECT NO. 505-0120 Table 7. Matrix Spike Recoveries in Rabbit senm (continued) 00050200002 (10050203003 . PFHA (ca acid) LL00 515.1 00050203006 00050203001? 1 00050203002. TDHA (C'I?acid) - LL00 303 00050203004 - - "000502030050 00050203006; 000502034302 . - 7 LL00 3.12-5me ch?qzn?m? . bin?Add. LL00=02?7ngihiL mama}, -- ?1me "03053203904. 78.3 Not Applicable 5 Page 9 of 18 IO 3M ENVIRONMENTAL LABORATORY PROJECT NO. EDS-0120 Table 7. Matrix Spike Recoveries in?Rabbit Sm?gqg?yuefi) ,7 . 00050203001] "c PFOA 0005:3203-003- 00050203005 000502037005 I 00050202001 r. A 4302 . 000502024003 ngImL; 06050202403} 000502024305 00050202 .Qc050202?001g- 00050202002 5 PFOA j' - LL00: 0.293 ngimL 00050202004 0 000502024305 FBSA 7_ 0 A .. 7 . 00050203005 . {00050203001 - '00050203-002' FOSA LLOQ 0.300 00050203000 - . 0 A 100% Page 10 oflS 3M ENVIRONMENTAL LABORA TORY PROJECT 20 Table-7. Matrix Spike Recoveries?in'Rabbit FFBS LLOQ 0.303 await. PFHs LLoq-s 0,315 'nglniL THPFOS LLog 0.780 nm: Tums mm 0.303?ngimLf ., lat-h. ,z-s .. QCDSOZDMOS Page 11 of18 1.2. 3M ENVIRONMENTAL LABORATORY PROJECT Table 8. PFOS Matrix Spike Recovery to Rabbit Serhm, Corrected for Endogenous Levels 00050202001 oco50202-002 (30050202003. 00050202-004 00050202005 .ocnsozdz-oos. .0 i 4. 3 Accuracy The accuracy for each analyte" reported was calculated by taking the average of the matrix spike recoveries for that analyte. The results are presented below. Table 9. Method Accuracy for Individual Analytes 17%: C12 Acid i 5% FBSA am . .. .7 FOSA, C7Acid? 7 43%, .PFBS 11% PFOA ,j 154% - ,izst cs Acid . 111% - PFOS C'1OAcid" - - THPFOS 212% 011mm use i192. "Thc Accuracy reported for this analytc is based on the high spike recovery data only because the low concentration spike recoueries were less than the LLOQ a 4. 4 Sample Related Comments The extraction and analysis of the samples was performed utilizing a non?Vahdated method. The matrix spike results, 13 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 01?18 15 3M ENVIRONMENTAL PROJECT Table 10. PerfluorinatedAcid Resultg mA'st' WA 6843 . TNA16847 6934 TNA 6935 TNA 6944 TNA 6959 NA 6960 TNA 6967 TNA 6973 TNA 6975 ?f Page 13 01?18 3M ENVIRONMENTAL LABORATORY PROJECT 20 151616119.- Perfluorinated Acid Results (691111116561 . . 5.04 41.00] 0.47? - .TNA 3961 6.73. - 2.45 6.00 7:41.00 "432 1.26 - (LCD 4.00 '1 21.001 TNA 6992. 0.690" 41.00; . $1.00 3 . 4.76 0.912 .. .1114 6994 .. 61.00 1.15: 0.337? 3 TNA 6995 16.41:? ?1621: 3.56 H.263 0.615 . 6997 3.90 41.00 3.48 2152.. 0.642' 4.00 TNA7000. 3.34 41.00 . 7 41.00 . 23.0.1? - 1103 . -