Document 0JavZxob2vBJ9xGjZEgRvq7Yx

! ::I I i P ..i:_i _:::! i.,..:'l !1 i iil !i:'i:_]'!: _iJ _i ._ I '_LJ ii:t: Study Title ANALYSISOF ENDOGENOUSFLUOROCHEMICALSINNORMALPOOLEDHUMAN SERUMAND Data Requirement 40 CFR Part 792 Author Mark E. Ellefson Study Completion Date 13 November, 2002 Revised Report 9 December, 2002 Performing Laboratory 3M Environmental Laboratory Building2-3E-09 935 Bush Avenue St.Paul,MN55106 Project Identification 3M Environmental Laboratory Study Number E02-1039 Total Number of Pages 225 3M ENVIRONMENTAL LABORATORY E02-1039 This page has been reserved for specific country requirements. I if'd !,,[ i,i;'! LL.i Page 2 of 225 ..;.!. i ,,:,,,_ !:;_ 'I 'i i 3M ENVIRONMENTAL LABORA TORY E02-1039 GLP COMPLIANCE STATEMENT Study Title: Analysis of Endogenous Fluorochemicals in Normal Pooled Human Serum and Plasma Study Identification Number: 3M Environmental Laboratory study Number E02-1039 This study was conducted in compliance with Toxic Substances Control Act (TSCA) Good Laboratory Practice (GLP) Standards, 40 CFR 792, with the exceptions listed below: Exceptions to GLP compliance: 40 CFR 792.130(e): There is not an electronic audit trail of corrections. The authenticated hardcopy printouts are considered the original raw data. mark E. Ellefson, Study Director //" "! ,l ', it 'i -WilliamK. Reagan, Testing Facility Management Date Date ,;.+.: .,,J i i/" i ,] Page 3 of 225 3M ENVIRONMENTAL LABORATORY E02-1039 QUALITY ASSURANCE STATEMENT ::i Study Title: Analysis of Endogenous Fluorochemicals in Normal Pooled Human Serum and Plasma Study Identification Number: 3M Environmental Laboratory Study Number E02-1039 This study was audited by the 3M Environmental Laboratory Quality Assurance Unit (QAU). as indicated in the following table. The findings were reported to the study director and laboratory i-ii_ management. ,,:,_,I i i : :! 1 10/10/02 10/11/02 10/29/02- 10/30/02 10/31/02 Protocol In-phase Data FinalReport 10/10/02 10/11/02 10/30/02 11/1/02 10/10/02 10/11/02 10/30/02 11/1/02 QAU Representative Date Page 4 of 225 3M ENVIRONMENTAL LABORATORY E02-1039 TABLE OF CONTENTS GLP Compliance Statement............................................................................................................... 3 Quality Assurance Statement ............................................................................................................ 4 List of Tables ...................................................................................................................................... 6 '7 I :i List of Figures ..................................................................................................................................... 6 Study Information ............................................................................................................................... 7 T!, Executive Summary ................................. ......................................................................................... 8 Summary ............................................................................................................................................ 9 Introduction.................................................................................................................................. ....... 11 i Test Substance .................................................................................................................................. 11 , Reference Substance ......................................................................................................................... 14 :_.i Test Systems ..................................................................................................................................... 15 Method Summaries ............................................................................................................................ 15 ; _! Preparatory and Analytical Method 15 Analytical Results ............................................................................................................................... 18 _ Data Summary ................................................................................................................................... 20 , Statistical Methods and Calculations ................................................................................................. 31 Statement of Conclusion .................................................................................................................... 32 References ......................................................................................................................................... 33 List of Attachments ............................................................................................................................. 33 i_!i.i_. i; Signature Page .................................................................................................................................. 34 Attachment A: Method ........................................................................................................................ 35 T!:', Attachment B: Data Tables ................................................................................................................ 56 _ Attachment C: Chromatograms .......................................................................................................... 63 Attachment D: Prep Sheets and Traceability Information .................................................................. 125 Attachment E: Protocol and Amendment ........................................................................................... 208 Page 5 of 225 ,_ ) L_ ' !i_I:,:_ ',,i_i_!'! 1t i I:L!2 ' ' 4 '.. t _i !._j! 3M ENVIRONMENTAL LABORATORY E02-1039 LIST OF TABLES Table l.a. Endogenous levels of test substance in normal pooled human serum ........................... 9 Table 1.b. Endogenous levels of test substance in normal pooled human Plasma ......................... 9 Table 2. Characterization of the Test Substances ............................................................................ 11 Table 3. Characterization of the Reference Substances .................................................................. 14 Table 4. Description of Test Systems Used in this Study ................................................................. 15 Table 5.a. Endogenous levels of test substance in normal pooled human serum ........................... 20 Table 5.b. Endogenous levels of test substance in normal pooled human Plasma ......................... 20 Table 6. Accurate Mass Determination of Endogenous Test Substances ....................................... 21 Table 7. Daughter Ions of Endogenous Test Substances ................................................................ 21 LIST OF FIGURES ' Figure 1: NMR Certified99.5% LinearIsomer PFOA and Mixed Branchedand Linear Isomer PFOA Standards................................................................................................................. 22 Figure2:C7 Isomer Distribution......................................................................................................... 23 Figure3:C8 Isomer Distribution......................................................................................................... 25 Figure4:C9 Isomer Distribution......................................................................................................... 26 Figure 5: Clo Isomer Distribution........................................................................................................27 Figure 6: Cll Isomer Distribution........................................................................................................28 Figure 7:C12 Isomer Distribution........................................................................................................ 29 Figure 8: PFOS Isomer Distribution ................................................................................................... 30 i_...J Page 6 of 225 , !,ii_ ;.S.,::i ..._. '.,i_:::r_ '! ! :I :! :.,,_,._ _:i_ , ..I I::7 i H',L::.t ii;_im_!l !_ . ;._ , .:.i '! ii;t STUDY INFORMATION 3M ENVIRONMENTAL LABORATORY E02-1039 Sponsor William K. Reagen 3M Environmental Laboratory Bldg. 2-3E-09 935 Bush Avenue St. Paul, MN 55106 651-778-6565 Study Director Mark E. Ellefson 3M Environmental Laboratory Bldg. 2-3E-09 935 Bush Avenue St. Paul, MN 55106 651-778-5405 Study Location Testing Facility 3M Environmental Laboratory Bldg. 2-3E-09 935 Bush Avenue St. Paul, MN 55106 - William K. Reagen, Laboratory Manager Stacy R. A. Hanson, Analytical Chemist Marlene M. Heying, Analytical Chemist Cindy M. Carlson, Analytical Chemist Ognjenka Krupljanin, Analytical Chemist Study Dates Study Initiation:10 October 2002 Experimental CInoitmiaptiolenti:o1n0: 3O1ctOocbteorb2e0r022002 Study Completion: 01 November 2002 Location of Archives All original raw data and the report have been archived at the 3M Environmental Laboratory according to 3M Standard Operating Procedures. Remaining specimens pertaining to the analytical phase of this study will be archived at 3M Environmental Laboratory for as long as the quality of the preparation affords evaluation. Page 7 of 225 3M ENVIRONMENTAL LABORATORY E02-1039 EXECUTIVE SUMMARY i :i A screening study of three lots of commercial pooled human plasma, one lot of pooled human , plasma from central China, and four lots of commercial pooled human serum was undertaken to quantify endogenous levels of perfluoroheptanoate (C7), pentadecafluorooctanoate (C8) (PFOA), ,._ heptadecafluorononanoate (C9), nonadecafluorodecanoate (C10), perfluoroundecanoate (C11 ), perfluorododecanoate (C12), and perfluorooctane sulfonate (PFOS). Results from this study showed quantifiable levels of the linear isomers of C7, C9-C12 ranging -i-t from < 0.010 - 0.9 ng/mL (Tables la and lb). Low to non-detect levels of branched isomers , were observed for the C7 and C9-C12 compounds detected (Figure 1 - 10). PFOS was present in the screened lots of commercial sera and plasma in concentrations ranging from 2.5 - 27 ng/mL and was present as branched and linear isomers. PFOA was present in the screened lots I of commercial sera and plasma in concentrations ranging from 0.65 - 5.6 ng/mL and was present i as branched and linear isomers. :i !i1 ",'1 J._J Page 8 of 225 r ; _I _:_ i _._,.; :,.':!' ..I._. ' __ :,,:i *_':_'i] !'_.._t _iil iJ , i_:,i 3M ENVIRONMENTALALBORATORY E02-1039 SUMMARY Quantitativescreeningswere conductedon four lotsof pooledhumanserumand four lotsof pooled humanplasma to determineendogenouslevels of perfluoroheptanoate(C7), pentadecafluorooctanoate(C8), heptadecafluorononanoate(C9), nonadecafluorodecanoate (C10), perfluorodecanoate (C11), perfluorododecanoate (C12), and perfluorooctane sulfonate (PFOS) as described by the 3M Environmental Laboratory Study #E02-1039 protocol. The analytical screening data is summarized in tables 1.a. and 1.b. Table1.a. Endogenoulsevelsoftestsubstancein normalpooledhumanserum Test Substance PFOS C12 Cll C10 09 C8 C7 :Sigma TCR-689 (nglmL) 4.56 [0.018] [0.076] [0.058] 0.265 1.60 [0.013] Lampare TCR-688 (nglmL) [2.49] [< 0.010]1 [< 0.010]1 [0.031] [0.068] 0.650 [0.025] uioresource TCR-687 (nglmL) 17.0 [0.144] 0.295 [0.327] 0.605 2.95 [< 0.010]' (3olOenwest TCR-690 (ng/mL) 27.0 [0.040] 0.320 0.203 0.900 5.60 0.190 [ ] Denotes values determined from concentrated spe method. _Value< LOQ. Tablel.b. Endogenoulsevelsof testsubstancein normalpooledhumanPlasma Analyte PFOS C12 O11 Clo 09 Cs C7 L;hlnese TCR-674 (ng/mL) < 1.01 < 0.101 < 0.101 < 0.101 < 0.251 < 0.521 < 0.10' Lampire TCR-685 (ng/mL) 11.1 [0.036] [0.049] [0.127] 0.435 2.61 0.100 Innovative Research TCR-683 (ng/mL) 15.6 [0.022] 0.135 0.170 0.585 3.07 [0.016] (5olOerlWest TCR-684 (ng/mL) 18,3 [0.024] [0.071] 0.160 0.535 3.87 0.290 [ ] Denotes values determined from concentrated spe method. 1Value< LOQ. Accurate mass measurements and elemental compositions were obtained for endogenous C7C11, and PFOS from a concentrated SPE extract of normal pooled human serum (see Table 6). Characteristic daughter ions were observed for endogenous C7-C12, and PFOS from a concentrated SPE extract of normal pooled human plasma (see Table 7). Qualitative analysis of chromatographically resolved branched and linear isomers of PFOA was accomplished using NMR certified standards of a linear isomer PFOA standard and of a mixed branched and linear isomer PFOA mixed standard. It was determined that the branched PFOA isomers elute within an approximate 0.2 minute retention time window and are baseJineresolved from the linear PFOA isomer. This finding of C8 isomer retention time resolution was Page 9 of 225 3M ENVIRONMENTAL LABORATORY E02-1039 extrapolated to determine qualitatively the presence or absence of branched isomers for the higher homologues of C9-C12, and PFOS (see Figures 1 - 8). i.i_ fRroemsu<lts0f.r0o1m0 -th0is.9stnugd/ymsLho(Twaebdleqsulaantaifniadblleb)l.evLeolswotfothneonli-ndeeater cistomleveerslsooffCb7r,anCc9h-eCd12isoramnegrisng were observed for the C7 and C9-C12 compounds detected (Figures 1 -8). PFOS was present U',f in the screened lots of commercial sera and plasma in concentrations ranging from 2.5 - 27 '_ ng/mL and was present as branched and linear isomers. PFOA was present in the screened lots i_,_I of commercial sera and plasma in concentrations ranging from 0.65 - 5.6 ng/mL and was present as branched and linear isomers. LJ L_J Page lO of 225 _:i ! _I _';i :_i -i J !'i:'!l ;!_'__ : _' ii .... 3 1- _it_ ,. _'l_i 3M ENVIRONMENTAL LABORATORY E02-1039 INTRODUCTION The purposeof this study is to perform quantitativescreeningfor perfluoroheptanoate(C7), pentadecafluorooctanoate(C8), heptadecafiuorononanoate(C9), nonadecafluorodecanoate (C10), perfluorodecanoate (C11), perfluorododecanoate (C12), and perfluorooctanesulfonate (PFOS) in normal pooled human serum and plasma. This study did not have a typical test substance that is dosed onto a specific controlled test system as per a conventional GLP study. The study design is in compliance with US EPA Toxic Substances Control Act (TSCA) 40 CFR Part 792. TEST SUBSTANCE Preliminaryscreeningindicatedthatthe test substanceslistedbeloware present as endogenous material in normalpooled human serumand plasma. Informationpertainingto traceability, source, physical description, and storage conditions is not available for these compounds as they exist in biological matrices. Table2. CharacterizatioonftheTestSubstances TEST SUBSTANCE FORM ILA Tridecafluoroheptanoate (C7) Pentadecafluorooctanoate (C8) Heptadecafluorononanoate (C9) Nonadecafluorodecanoate (C10) Perfluoroundecanoate C11) Perfluorododecanoate (C12) Perfluorooctane sulfonate (PFOS) C_F_COO" CzF,sCOO CsF_COO" _F_oCOO" C_oF2_COO C.Fz3COO CsF,TSO_ The molecular structures are given below. Name: C7 Chemical Molecular Name: Perfluoroheptanoate Weight: 363, as shown F F F O F F_@@_0 - F F F Page 11 of 225 ,.'1 i : ; _.A _.L., I -i _' !:j ::, _,, Lr_ :i :':_: } -: 3M ENVIRONMENTAL LABORATORY E02-1039 Name: C8 Chemical Molecular Name: Pentadecafluorooctanoate Weight: 413, as shown F F F 0 F F_O F F F F Name: C9 Chemical Molecular Name: Heptadecafluorononanoate Weight: 463, as shown F F F F O Name: O- F F F F F C10 Chemical Name: Nonadecafluorodecanoate Molecular Weight: 513, as shown F F F F O F_ o- F F F F F Name: C11 Chemical Molecular Name: Perfluoroundecanoate Weight: 563, as shown F F F F F O F 0- F F F F F Page 12 of 225 3M ENVIRONMENTALLABORATORY E02-1039 Name: C12 Chemical Name: Perfluorododecanoate Molecular Weight: 613, as shown ;'<i F F F F F 0 F 0- .i ] F_ ' F F F F F F Name: PFOS Chemical Name: Perfluorooctanesulfonate !,,q Molecular Weight: 499, as shown F F F F /F/F/F tF _ ] .,. F F F F Page 13 of 225 "i , ! ,:"i_i i, i :f_ I '! ' "'_ ,,:,;::,i _ i iiJ i REFERENCE SUBSTANCE Table 3. Characterization of the Reference Substances 3M ENVIRONMENTAL LABORATORY E02-1039 Tridecafluoro heptanioc Acid Pentadecafluoro octanoic Acid Heptadecafluoro nonanoic Acid Nonadecafluoro decanoic Acid Perfluoroun decanoic Acid Perfluorodo decanoic Acid C6F_3COOH CTF15COOH TCR-267 TCR-617 CsFlzCOOH TCR-618 CgF_gCOOH TCR-036 CIoFzlCOOH TCR-619 C,Fz_COOH TCR-037 Aldrich Oakwood Products Oakwood Products Oakwood Products Oakwood Products Oakwood Products Potassium Perfluorooctane sulfonate 08F17SO3K* TCR-018 SMM* 236-113-10 *Documentation of the method of synthesis is located at the source. confirmed isomers **Reference substances as linear by NMR. Clear Crystals White Crystals White Crystals White Solid White Crystals White Powder White Powder 98.2%** 99.5%** 98.02%** 98%** 96.4%** 99.7%** 86.9% Frozen Ambient Temperature Ambient Temperature Frozen Ambient Temperature Frozen Frozen .... J Page 14 of 225 r 'i i__ :-i ' !ii_?l 4, _ _1 ' :t ::J r= : :_ i _.,i_;_ !_:_:i_ _il _ !i!!_],.!_._ , _; i._..;.i.i TEST SYSTEMS Table4. Descriptionof TestSystemsUsedin thisStudy 3M ENVIRONMENTAL LABORA TORY E02-1039 Pooled Human Serum PooledHuman Serum PooledHuman Serum Pooled Human Serum Pooled Human Plasma Pooled Human Plasma Pooled Human Plasma PooledHuman Plasma Sigma-AIddchM,ilwaukeeW, l LampireBiologicaLlaboratoriesP,ipersvilleP, A BioresourcTeechnologyI,nc.,FortLauderdaleF,L GoldenWestBiologicalsT,emeculaC, A LampireBiologicaLl aboratoriesP,ipersvilleP, A GoldenWestBiologicalsT,emeculaC, A InnovativeResearchI,nc.,SouthieldM, I CentraCl hina TCR-689L, ot022K0965 TCR-688,LotX324B TCR-687,Lot020821 TCR-690,LotG01406042 TCR-685L, ot22-60824A TCR-684,LotG01410002 TCR-683,LotIR02-014 TCR-874,LotN087P27 Justification of the Test System. Based on preliminary testing, normal pooled serum and plasma contain endogenous levels of the test substances. Identification of Test System. Samples were identified by the TCR number. Collection of Test Systems. The commercial vendors indicated that three of the four lots of commercial pooled human serum and three lots of commercial pooled human plasma purchased for this study were collected from subjects residing in close proximity to the individualvendors (Sigma-Aldrich purchases blood products from blood banks located throughout the US). Hence, although limited to single lots of serum or plasma per location, analytical data from the purchased serum and plasma provide a benchmark of endogenous levels of fluorochemicals from different regions of the country. METHOD SUMMARIES Preparatory and Analytical Method ETS-8-231.1, "Solid Phase Extraction and Analysis of Fluorochemical Compounds from Biological Matricies." A 2.0 mL aliquot of serum or plasma is transferred to a 50 mL screw- capped polyethylene centrifuge tube. Spiking solution is added as appropriate, followed by the addition of 8.0 mL of ASTM Type I water. The mixture is shaken and 40.0 mL of acetonitrile are added. The sample container was capped, mixed for 20 minutes, and centrifuged at 3,500 rpm for 20 minutes to clarify the supernatant. Following centrifugation, the supernatant is transferred to a 500 mL Nalgene container, diluted with 350 mL ASTM Type I water, and passed through a pre-conditioned Cla solid phase extraction (SPE) cartridge. The analytes of interest are eluted from the SPE cartridge using 2.0 mL of methanol. Concentrated SPE Method, A modified form of the SPE method described by ETS-8-231.1, "Solid Phase Extraction and Analysis of Fluorochemical Compounds from Biological Matricies." used to concentrate analytes present at concentrations below the LOQ. A fivefold concentration is achieved by using 5 times more serum or plasma while maintaining the foinr apllaeslumtiaontravnoslufemreredat t2o.0a m25L0mmeLthNanaolgl.enTehe cmoentthaoinderc.onSspisiktsingofsaol1u0ti.o0nmwLasalaiqdudoetdofasserum appropriate, followed by the addition of 40.0 mL of ASTM Type I water. The mixture is shaken and 200.0 mL of acetonitrile are added. The sample container is capped, mixed for 20 minutes, and centrifuged at 3,500 rpm for 20 minutes to clarify the supernatant. Following centrifugation, the supernatant is transferred to a 2.0 L Nalgene container, diluted with 1750 mL ASTM Type I water, and passed through a pre-conditioned C_8solid phase extraction Page 15 of 225 '. :'.! _i ,, _ _--_ _i ii,,i _t i! _' I ! L-| L_'t ;T i 'i i _:'__ 3M ENVIRONMENTAL LABORATORY E02-1039 (SPE) cartridge. The analytes of interest are eluted from the SPE cartridge using 2.0 mL of methanol. Further sensitivity was achieved by doubling the injection volume of the standards used to calibrate the run. Analytical Method Liquid Chromatograph: Hewlett-PackardSeries 1100 Liquid Chromatograph Analytical column: KeystoneBetasilTMC182xl00mm, 51Jmparticle size Column temperature: 40 C Cycle Time: 17.0 minutes Flow rate: 300pL/min Injection volume: 10 pL (20 IJLfor concentrated SPE method only) Mobile phase components: Solvent A: 2.0 mM ammonium acetate in water Solvent B: Methanol system Solvent Gradient: Time (rain.) 0.00 10.00 11.00 11.50 12.50 13.00 16.00 %B 40% 90% 90 % 100 % 100 % 40 % 4O % Mass Spectrometer: Micromass Quatro Ultima triple quadrupole mass spectrometer Software: Masslynx version 3.5. Mass Spectrometer Acquisition Parameters: (c7) Channel Parent Ion Daughter Ion Collision Cone (V) (m/z) 1 363.00 (m/z) Ener.qv {eV) 119.00 20 20 2 3 4 (c.) Channel 1 2 3 4 363.00 363.00 363.00 Parent Ion (m/z} 413.00 413.00 413.00 413.00 169.00 20 219.00 20 319.00 20 Daughter Ion (m/z) 119.00 169.00 219.00 369.00 Collision Enerqv (eV) 20 20 20 20 20 20 20 Cone (V) 20 20 20 20 Page 16 of 225 ! _i !, I .! , -! I I1 i_ill _ii_' :_ '- i I i ii, (C9) Channel 1 2 3 4 (Ct0) Channel 1 2 Parent Ion (m/z) 463.00 463.00 463.00 463.00 Parent Ion (m/z) 513.00 513.00 Daughter Ion (m/z) 119.00 169.00 219.00 419.00 Daughter Ion (m/z) 119.00 169.00 3M ENVIRONMENTAL LABORATORY E02-1039 Collision EnemY (eV) 20 20 20 20 Cone (V) 20 20 20 20 Collision Enerqy (eV) 20 20 Cone (V) 20 20 4 (Ctl) Channel 1 2 3 4 513.00 Parent Ion (m/z) 563.00 563.00 563.00 563.00 469.00 20 Daughter Ion (m/z) 119.00 169.00 219.00 519.00 Collision Ener.qv(eV) 20 20 20 20 20 Cone (V) 20 20 20 20 Channel 1 2 3 Parent Ion (m/z,) 613.00 613.00 613.00 Daughter Ion (m/z) 119.00 169.00 219.00 Collision Enerqv (eV) 20 20 20 4 613.00 569.00 20 Cone (V) 20 20 20 20 Page 17 of 225 'ii ii i i'] :i !"'/!_'_\_:-ii irU"I _:'_ _,_,, _ :__ i i :i:, i_i_ 3M ENVIRONMENTAL LABORA TORY E02-1039 PFOS Channel 2 3 Parent Ion (m/z) Daughter Ion (m/z) ,0000 000 499.00 99.00 Collision Enerqv {eV) 45 499.00 130.00 45 Cone (V) 00 60 60 Capillary Voltage: 4000 V Gain = 1.0 EMV Mode: Electrospray Negative Gas Temperature: 250 C DNreybinuglizGerasP:re8s.0suLr/em: i3n0. psig Analysis Type: Multiple Reaction Monitoring (MRM) ANALYTICAL RESULTS Data quality objectives outlined in the 3M Environmental Laboratory method were met (see Appendix A). Regressions. Quadratic curve fit weighted 1/x was applied to calibration standards and sample data to improve quantitation over the concentration range appropriate to the data. All calibration curves had a coefficient of determination of 0.998 or greater. Calibration Standards. Calibration curves were prepared from extracted matrix standards in Chinese plasma for all plasma and serum quantitations. Reported values were not corrected for endogenous levels of test substance in the Chinese plasma calibration curve (levels of all endogenous test substances in the Chinese plasma were determined to be < LOQ). The curves a points. by regresscioonnsaisntaeldysoisf usminingimthuempeoafknainreea(9s)of the anTahlyetee.qTuhaetioancwcuarsacdyeotefremaicnhedlevelwas verified. Any level outside70% - 130% of nominalwas deactivated,and regression re- ConctianluciunlgateCda. liAblrlalteiovnelsVsehroifwiceadtioanr.espFoonrsqeuagnretitaatteivrethdaentetwrmicienathtiaotnosf, tahembidla-lnekve. l matrix calibration check was analyzed at least every ten samples to monitor instrumental drift, with a limit of +25% deviation of the target concentrations. Limit of Quantitation (LOQ). The LOQ was equalto the loweststandardin the calibration curve,with a level of accuracywithin+30%. The levelof analyte in the LOQ was also greater than two times the response of analyte in the blank samples. Demonstration of Specificity. The identification of analytes was substantiated by chromatographic retention times, characteristic primary ions, characteristic daughter ions, and isomeric proportions (where applicable). Control of Bias. Two levels of matrix fortifications, prepared at knownconcentrations of the test substance and bracl_eting the anticipated range of the method were evaluated to determine recovery and to evaluate method performance. In serum samples, PFOS and C12 exhibited matrix effects identified by high (>150% recovery) or low (< 75% recovery. For all other analytes, and for all analytes in plasma samples, recoveries ranged from 75% to 105%. Reagent and matrix blanks were run with each set to evaluate the level of background interferences. Page 18 of 225 3M ENWRONMENTAL LABORA TORY E02-1039 Blanks. Method blanks and matrix blanks were evaluated in the course of this study. The method blanks showed no evidence of background contamination introduced in the sample ,, preparation stage. The matrix blanks did show endogenous levels of the analytes. This was ,:,"4 expected and such instances were noted in the raw data. ,I I_'_, J i ii Page 19 of 225 t1 :,!] , ,_ ._ 'i ;! ! _ :_'i_ i; -_i i !!_I!': ;_ __1,i_;' _..._ .i , i__,_ 3M ENVIRONMENTAL LABORATORY E02-1039 DATA SUMMARY Quantitative screenings were conducted on four lots of normal pooled human serum and four lots of normal pooled human plasma for the determination of endogenous levels of perfluoroheptanoate (C7), pentadecafluorooctanoate (C8), heptadecafluorononanoate (C9), nonadecafluorodecanoate (C10), perfluorodecanoate (C11), perfluorododecanoate (C12), and perfluorooctane sulfonate (PFOS) as described by the 3M Environmental Laboratory Study #E021039 protocol. The screening data is summarized in tables 5.a. and 5.b. Matrix spike recovery information is contained in Appendix B. Table 5.a. Endogenous levels of test substance in normal pooled human serum Test Substance PFOS O12 CtI C10 C9 C8 07 _51gma TCR-689 (ng/mL) 4.56 [0.018] [0.076] [0.058] 0.265 1.60 [0.013] Lamplre TCR-688 (ng/mL) [2.49] [< 0.010] 1 [< 0.010]1 [0.031] [0.068] 0.650 [0.025] Bloresource TCR-687 (ng/mL) 17.0 [0.144] 0.295 [0.327] 0.605 2.95 [< 0.010]' (5olden west TCR-690 (ng/mL) 27.0 [0.040] 0.320 0.203 0.900 5.60 0.190 [ ] Denotes values determined from concentrated spe method. 1Value < LOQ. Table 5.b. Endogenous levels of test substance in normal pooled human Plasma Analyte PFOS 012 unmese TCR-674 (nglmL) < 1.01 < 0.101 Lamplre TCR-685 (nglmL) 11.1 [0.036] Innovative Research TCR-683 (ng/mL) 15.6 [0.022] _olden west TCR-684 (ng/mL) 18.3 [0.024] 0C1 1t0 C9 C8 07 '<< 00.1.01101 < 0.251 < 0.521 < 0.10' [00..014297] ] 0.435 2.61 0.100 00.1.315 70 0.585 3.07 [0.016] [ ] Denotes values determined from concentrated spe method. 1Value< LOQ. [0.107610] 0.535 3.87 0.290 Accurate Mass Determination, Accurate mass measurements and elemental compositions were obtained for endogenous C7-C11 and PFOS in a concentrated SPE extract of normal pooled human serum purchased from Golden West Biologicals (TCR-690). Accurate mass measurements are presented in Table 6. Page 20 of 225 "I ,:_! Ji _:i ""; L) , iI ! I i/_:_i _.! i::i:.:'i! , 3M ENVIRONMENTAL LABORA TORY E02-1039 Table 6. Accurate Mass Determination of Endogenous Test Substances Test Substance Accurate Mass Theoretical Mass C7 C8 C9 PFOS C l0 C 11 362.9739 412.9637 462.9613 498.9280 512.9615 562.9542 362.9691 412.9659 463.9627 498.9297 512.9595 562.9563 Mass Deviation (ppm) 13.3 -5.4 -3.0 -3.3 3.9 -3.8 Probable Formula C702F13 C802F15 C902F17 C803F17S C loOzF 19 CI IO2F21 Daughter Ions of Endogenous Test Substances, Specificdaughter ionswere observedat characteristic retention times for endogenous C7-C11 and PFOS in a concentrated SPE extract of normal pooled human plasma purchased from Golden West Biologicals (TCR-684). Daughter ions verified in analytical standards of linear isomers. A summary of daughter ions observed is presented in Table 7. Table 7. Daughter Ions of Endogenous Test Substances Test Substance Cr C8 C9 C,0 Cll 612 PFOS Parent Ion (m/z) 363 413 463 513 563 613 Dau_lhter Ions (m/z) 319 119 369 219, 169, 119 419 269, 219, 169, 119 469 319, 269, 219, 169 519 319, 219, 169, 119 569 419, 319, 269, 219, 169, 119 499 130 99, 80 Iesnodmogeersn,oTushefluporeroscehnecmeicoar lasbwseenreceqoufalditiasttirvibeulytiodnesteormf binraendcihnecdoanncdenlitnraetaerdisSoPmEeresxtorfacts of commercial lots of pooled human serum and plasma (Figures 1 - 10). Qualitative analysis of chromatographically resolved branched and linear isomers of PFOA was accomplished using NMR certified standards of a linear isomer PFOA standard and a mixed branched and linear isomer PFOA standard. It was determined that the branched PFOA isomers elute within an approximate 0.2 minute retention time window and are baseline resolved from the linear PFOA isomer. This finding of C8 retention time resolution was extrapolated to determine qualitatively the presence of branched and linear isomers for C7, the higher homologues of C9-C12, and PFOS. Low to non-detect levels of branched isomers were observed for the C7 and C9-C12 compounds detected. PFOS and PFOA were present in the screened lots of sera and plasma as branched and linear isomer distributions. Figure 1 contains chromatograms of NMR certified linear isomer PFOA and mixed branched and linear isomer PFOA standards. Figures 2 - 8 contain chromatograms of the corresponding standards of C7 -C12 and PFOS in solvent, an extracted calibration curve point (extracted calibration curve point consists of pooled human plasma from central China spiked with 10 ng of test substance/mL of plasma), and three concentrated SPE Page 21 of 225 3M ENVIRONMENTAL LABORATORY E02-1039 extracts of commercial pooled human serum. Pooled human plasma collected in central China was used as the blank matrix to construct an extracted calibration curve because preliminary screening results indicated it did not contain quantifiable levels of endogenous test substances. :' TI &_,,, ,"- .3 i :1 iJ 4 ,2:J i ::'2'J Page 22 of 225 _,,-', j::,'_')::!:i 3M ENVIRONMENTAL LABORATORY E02-1039 Figure 1: NMR Certified 99.5% Linear Isomer PFOA and Mixed Branched and Linear Isomer PFOA Standards d021 106002 Sm (Mn, lxl) 1 00- Unear k omer C8 standard 8.43 51249- <= linear (gg.5% MRM of 4 Channels ES- by" NMR) TI C 3.7 7e5 Area %. Lt_} 8.19 ...._i branched(0.5% byNMR)=> 503___ 0 ':'T'li" '_', .... ',...'. _... ;i..-.;'i" , .... i .... i .... i .... i:.'"-";-I '-;'ff_"'' d021106001 Sm (Mn, lxl) 1 00- 8.43 153171- MRM of 4 Channels E3TI C <= linear(77.7%by NMR) 1.35e6 Area !_'_ Branched & linear isomer PFOAstandard _ ! ,i i;;:,j' "._-_ _-i =%. 8.18 50677 7 88 0- Time Summary of branched and isomer PFOA 2.50 5.00 7.50 1 0.00 PFOA Standards: NMR certified standard of the linear 99.5% linear isomers. NMR certified standard of mixed contains 22.3% branched and 77.7% linear isomers. 12.50 15.00 isomer PFOA contains branched and linear 0.5% Page 23 of 225 Figure 2:C7 Isomer Distribution 3M ENVIRONMENTAL LABORA TORY E02-1039 ,r _,,t -_i_ i 1 :"_ '.L } _[ ;} ; _"_" . ' i ...:. d021025069 Sm (Mn, lxl) 7 3 __ <= Unear 1: MRM of 4 Channels ES. TIC Solvent standard 30 ppb 7.1 | 116_:| ranohed=, 1(_1 i i i i i i i i J i i i i i | ! i 2 678mj, | I l i l i i I I l l I I I ! 11 ,,1 d021025091Sm (Mn, lxl) 7.3 Chines e plasma 1Oppb Area 2.09e5 1 | l I l i i I I I I I I i i i i i I ! i I | o 1: MRM of 4 ChannelsES. 2.66e5 Area 1 I1 (branched not obse2rv6e2.d9) .1i <=- Un. ear . d021025098m (Mn, lxl) Lampireserurn _ --7_ < Unear (branched not observed) IfLI1 111 .... I .... I .... I .... I .... I 1..../i_, I /..I.. I .... I .... d021025108 m (Mn, lxl) S a serum - 7 2 <- linear I, (branched notobserveoj _=_,- j. TIC 1: MRM of 4 ChannelsES. 2.g8e4 Area I .... I .... I .... I' '' TIC 1" MRM of 4 Channels ES. .32e4 rea d0210in25ill 10,2, S,,mi,,(Mn,i , lljxl),, _i ,in =i ,_ i, l,i ,, i= Ii, i, ,i ,|, , 1=,i MiR= M=, ofill 4i=Chli a,n'nels ES- TIC Golden West serum Area 11%_ _'_54 <= Unear /_1 4.93e4 (branched not observed) "lJ_ ._ L it,,_, OJ'-.1,, .... ,,. : ,._, .',',":'_..'_,'."_...., 2.50 5.00 7.50 _','.":_,1"_,,",/'_/._,,, :.-:_,.,. ,-'-"T--im='e, 10.00 12.50 15.00 Summary of Cz Acid Chromatograms: Low to non-detect levels of the branched isomers of Cz acid were observed in these lots of Commercialpooled human serum. Page 24 of 225 3M ENVIRONMENTAL LABORATORY E02-1039 ,i % Lb i; -. "';:_'::_ _,, i i ::.A , ..... b:."- _:i!_ _,_ _,";!;_ i_; ; i ....; iJ,::} Figure 3: Ca Isomer Distribution d021024012Sm(Mn, lxl) Sohtentstandard 30 ppb <= Linear 2: MRMof 4Channels ESTIC 3 23e5 Area 8.22! === ii l= ==1 i= ii ill Branched => 48tl == i= i= =1 =l ill ii =l d021024034Sm(Mn, lxl) B.47 69023 Chinese plasma lOppb ll= == i= ill <=Unear i= ii ii li=ll ,= l= ill == 2: MRMof 4ChannelsESTI C 6.80e5 Area 8.21i Branched => 534| "%1 0" .... = .... = .... i .... i .... i .... , .... , .... i .... i .... i .... t .... i .... d021024041Sm (Mn,11) 2: MRMof 4 Channels ES- 100-_ Lampire serum / 683.64045q1 <=Linear 6.3T0eIC5 Area "1 Branche=d> 180.2118_|1 0 / _ , i , _ _ _ i ' _ J = i I _ ' I I , , M ' i =% ' ' i '%' * = 1 _ = ' ' i ' = _ '*1 _ _ J _ i ' = = _ I _ = _ J I = , , _ d021024043Sm (Mn,lxl) 2: MRMof 4 Channels ES- Sigma serum 79360 <= IJnear 8.1 4 / 1_1 Oi'" "' I"'' Branched => 87.2458_i1tI "i "" "'" I '' "--'%"1 .... I .... I .... i ..... I .... d021024045m(Mn, lxl) 8.40 8.1 385 Area i .... I .... "I''-'_T TIC " :'-" 2: MRMof 4ChannelsESTIC Golden west serum Area 0" .... i .... , .... 2.50 I 7.aO Branched=> 861.91 , .... , .... , .... ,_'", 5.00 7.50 '", .... 10.00 , .... , .... l .... , .... Time 12.50 15.00 Summaryof C8Chromatograms:Lowto non-detectlevelsof branchedisomersof C8were observedin theselotsof commercialpooledhumanserum. Page 25 of 225 ; ! :_!7 ', ;!:':,::i iI ::J i. I ":_' .... "_ i '-'_ 't i! 3M ENVIRONMENTAL LABORATORY E02-1039 Figure 4:C9 Isomer Distribution d021025069 Srn (Mn, lxl) Solventstandard 30 ppb 40551 <= Unear 2: MRN of 4 Channels ES. 4.0105 Area (branched not observed) .... ! .... I .... I .... I .... d021025091 Srn (Mn, lxl) I' '' '1 .... I .... 0.1o I ' '' 't .... I .... I '''' I .... 2: MRM of 4 Channels ETIC C hinese plasma lOppb (branched not o_erved) .... t .... i .... I .... I .... i .... i .... d021025098 m (Mn, lxl) 1<= Linear i .... , .... f .... , .... I ''' '_-_1.... Area 2: MRM of 4 ChannelsES. La mp ire se rum _50 t_, _ 888 I <= Unear Area 0"-1,. :_._.... , .... ,...',...-:,.7.., d021025100 Sm (Mn, lxl) Sigma serurn : .... :','. , .................... 2: MRM of 4 Channels ES. TIC g.04 7041 < Area _- _ ' _ t" * a _ I ' " ' ' I ' ' '' I _ _B_ r_anI c'-_he_d=' >I ' _ J_ ,'_, I ' ' ' _t _ ' _ ' I I-'" "'_t_'-t _' ' '' d021025102 8m (Mn, lx1) Golden wests erum 004 1_24 2: MRM of 4 ChannelsTEISC. I 2.8605 Area 0 I' ' '--;-_f" ' ' ' I .... 2.50 188_t_I <=Linear J[ Branched=> '11 _ .- It 1% ' ' ' I ' ; ' '' I ...." "!_" .... "_I .... I _.."._. "*'-_"--_=" .... I ...%. --'_"_1'_I .... I ' ' ' T/IIIIU_._ 5.00 7.50 10.00 12.50 15.00 Summaryof C9Chromatoqrams:Lowto non-detectlevelsof branchedisomersof C9were observedin theselots of commercialpooledhumanserum. Page 26 of 225 __ Figure 5: Clo Isomer Distribution 3M ENVIRONMENTAL LABORATORY E02-1039 ::_3 i_'i i .+: _!_,:_ '_:"," _%i ..."... i:!J i:_.! '! ,_ I _f_l +.._..':_'_ '! +: --_' _:_ !'i.!_! d021024012 Sm (t,,,lnl,xl) 1007 Solvent standard "1 38 ppb d021024034 Sm (Mn, lxl) 100- 9.91 _ 28022 3: _IRMof 4 Channels ES- TIC <=Linear 2.A87reea5 Branched => _. 930..7_/7 9.89 46914-j 3: MRMof 4 Channels ESTI C <=Linear 4.97e5 %- Chinese plasma lOppb O' ""'l .... i .... f .... d021 124041Sm (Mn, lxl) 100-_ Lampire serum "l 0 [I '' II ''= '1 '= ''1 =' '' d021024043 Sm (Mn, lxl) i .... I= '''Ill= [ 9B3/ Area era,-,o=h>edzo3| +.... I .... I .... +.... , .... J.... ? .... I .... 3: MRMol' 4 Channels ES- 9.88 TI C 1745 qI, 9.701 . <= Unear 2.A00reea4 Branched => 104 '1 =l ''i II '= I= =l' 9.84 I' 'l =1 '= ==1 '' 'l I+' '' 3: MRMof 4 Channels ESTI C 1001 Sigma serum 2457"1_ <= Linear 2.A88reea4 II %_ = 9.691 0 ''_ ,+_'--:"'-;-" "--, '-' - , .... , .... , .... , .... i .... , .... , .... , .... d021024045Sm (Mn, lxl) 3: MRMo14' ChannelsES- 9.84 TI C 9223 1.00e5 l<=.+r Golden west serum 9.66 I Branche=d> 10.86 12.30 0 . . .'., ,., ,'_q_',. .-_-. ,-, .... __, ._..... , .... , .... , .... , ....9_,4...9-__. 2.50 5.00 7.50 10.00 12.50 15.00 Area Time Summaryof C_o,ChromatogramsL:owto non-detectlevelsof branchedisomersof C10 were observedin theselots of commercialpooledhumanserum. Page 27 of 225 i,il,i p i ,i ,,,I,j ...._ :"] _,i : !:_i1 .... _iit _ 3M ENVIRONMENTAL LABORATORY E02-1039 Figure 6: Cll Isomer Distribution ct021025069 Sm (t_ln1, 1) 100] 10.31 54214 q 3 MRM of 4 Channels EBTI C 5.7 2e 5 Solventstandard30 ppb i "1 0""_ u u-, j , a u, j , , , u-j u'_ , , | .... j;3 d021 025091 Sm (Mn, lxl) 1 00_ _n ChineseplasmaIOppb i 1o.o7/ lO31 II <= Unear Area , , lu B, r,annj c="h, _e-,dj=,>= = ui-,-, u_, "_=jI,,, , , uTT]TT,-,-u j "g-=a n , i u_r'l' , = u 10.28 _ 43258 3: MRM of 4 Channels ES- TI C 4.54e5 <= Linear Area 96t 0 / .... ,'I''1 Branched=> 703 10.05 .... , .... I .... | .... I .... I .... I .... I ....... I .... I .... I .... I .... , .... d021 02509fl Sm (Mn, lxl) 1 00q- 10.31 542 -_ 3: MRM ef 4 Channels ESTI C 7 44e3 _ Lampireserum = I <= Linear Area d021 0251 00 Sm (Mn, lxl) 100 _ _] Sigmas erum t% 0 _-.;., .... i .... , .... , .... ,'" 10.23 3: MRM of 4 Channels ESTIC "1 3318| <= Linear 3.75e4 Area Branched=> 10.0711 358,_1 , .... ,...-, :.'-t-'. ;., .... ,.... , .... ,..*"_.1.... _'.... d021 025102 Bm (Mn, lxl) 100-_ 10.23 8903 "-_ 3: MtRM of 4 Channels ETSI-C 9.71e4 _= linear ,Area Goldenwest serum 0J .... , .... i .... , .... , .... 2.00 4.00 114111 10.07| Branched=> '_ , .... , .... , .... i .... , .... I .... 8.00 8.00 10.00 , .... , .... 12.00 Summary of C,11Chromatograms: Low to non-detect levels observed in these lots of commercial pooled human serum. | .... | .... , ..... Time 14.00 of branched isomers of C_ were Page 28 of 225 _,,_, ...!.. _iJ _' .{:', "_ 'i ii,_.i! _!i i ..i 3M ENVIRONMENTAL LABORATORY E02-1039 Figure 7: Clz Isomer Distribution d021025126 Sm (Mn, lxl) 10 Solvent standard 30 ppb 10.75 o2a2e 2: MRM or 4 ChannelsES. TIC <= Unear 8.0005 Area (branched not observed) ' = '' I ' ' ' " I '' ' ' I' " '' I ' '' ' I ' ' ' 'I ' " ' ' I" " ' ' I ' '' ' I '' t' ' ' ' I ' ' ' ' I ' w '= d021025148 Sm (Mn, lxl) lO.72 2: MRM of 4 Channels ES. TIC Chinese plasma 10ppb (branched not observed) ! ,,,', =.... i r,.,, ; r,, r ,-.;,-i_,, _,, ;;, =, ,-,; i;,,, d021025155 Sm (Mn, lxl) 1007 1 Lampire serum ] 10.75307 ] (branched not observed) J ,_-_i;;,, 2: MRM <= Linear I T;-,-.,-i;,, of 4 Channels ES. Ic 6.9T6p.3. Area o ,', ,, ,, , , , .,, , w ,, o., ,, i, . ,=,1, d021025157 Sm (Mn, lxl) 1 n_ Olu_ ,, , , , ,,,. , , , , i, , lO.72 528 "i _) , , ,, . , , , , ., , , , , , ,, 2: MRM of 4 Channels E. <= Unear TIC 1.01 e4 _ Sigmaseru m ,Zl I Area 0"_ .... I ' ' ' ' I .... I .... I ' '' ' I .... I .... I .... I .... I ' ' '' [' ' ' ' I ' ' ' ' I' ' '' d021025159 Sm (Mn, lxl) lO.71 2: MRM of 4 Channels TEICS. lo.m A rea 1_1 ,G_ol-den wesi,t.,s-,e-rju,5m_ ,.............,...... , .... , ...1. 478, ..Q.. , <..=.. lin,e.a..r. , . ,'. 2T.3im50e4 2.58 5.00 7.50 10.00 12.50 15.00 Summary of C_ Chromatograms: Low to non-detect levels of branched isomers of 012 were observed in these lots of commercial pooled human serum. Page 29 of 225 " -1 ii"!1_ i;'_'1 ! :::i:_ ; i i!_i':i 7:;} .j !] ] ; _! _ ,.:L "' :i Figure8: PFOSIsomerDistribution 3M ENVIRONMENTAL LABORATORY E02-1039 d021lI25126 Srn (ran, lxl) 100- Solvent standard 38 ppb 9.08 7 85187l I B.BI I 50373 / <= Unear 1. M_M of 3 Channels ESTIC 9.15e5 Area 0 .... I .... I .... I .... I .... d0211125M8 Srn (Mn, 1)1) 100- Chinese plasma 10pph o .............. dO21(25155 Sm (Mn, lxl) 1go- , ......... Branched => I .... I .... I .... , ..................... 378B5_ I .... I .... I .... I .... I .... I .... I .... I .... 590.70974_/ i i 8.79 | <= Linear 1: MRM or 3 Channels ES6.64T0I5C Area 8.55 228 9.09 _ 94485 I ,....... <= Linear , ........................ 1: MRM of 3 Channels ESTIC 1.2385 " Lamp|re serum Branched => ii 785158|813' 8.57 Area O ' ' ' ' I .... I .... i .... I .... I .... I .... I .... T' ' '_' I .... I .... I .... I .... I .... I .... t .... C10211)25157Sm (Mn, lxl) 1: MF_Mof 3 Channels ES- 9.05 TIC Sigma serum '!1" ' ' ' ' I .... I .... I .... I .... {3021ll25159 Sm (Mn, lxl) 10O- %- Goldenwest serum " 0 , , ...,...,.'_..'.....,..,. . .... 2.00 ,, _ 4.OO Branched=> I .... l .... I .... 18e363 / tl 851 21 |0q<8:7n7ear 514oj| _ .... I ''_ ' ' I .... I .... I .... 9.04 707835] 5584.73899 II <= Unear 8.52II 18417 , B, r,a,n.c..h. ed i: ) ; _" _1 _ ......,..............L...... , 6.00 800 1000 12.00 I .... I .... I .... ,,e Area 1: MRM ot 3 Channels ESTIC 867e6 Area , ; , 14.00 Time Summary of PFOS Chromatograms: Branched isomers of PFOS were observed in these lots of commercial pooled human serum. Page 30 of 225 i_!ii '_! I !:!,,,] _i[_ ii!1i _i] 3M ENVIRONMENTAL LABORA TORY E02-1039 STATISTICAL METHODS AND CALCULATIONS Theoretical concentrations of analyte in final eluate: Concentration = (Concentration of Analytical Standard x Amount of standard added) / EV Observed result to original sample result: Original sample result = Observed result x DF Spike percent recoveries: %Recovery Observed Result - Same Sample, Unspiked = Theoretical Concentration x 100 EV= Eluate volume DF = Dilution factor Relative standard deviation: Standard Deviation Relative Standard Deviation = x 100 Mean % Deviation Means and standard = Theoretical TheCoornetci.ca-l CCalocnucla. ted Conc. x 100 deviations were calculated using Excel software. i :I I? i Page 31 of 225 3M ENVIRONMENTAL LABORATORY E02-1039 STATEMENT OF CONCLUSION Resultsfrom this study showed quantifiablelevels of the linear isomers of C7, C9-C12 ranging from < 0.010 - 0.9 ng/mL (Tablesla and lb), Low to non-detect levelsof branched isomers :_"_ were observed for the C7 and C9-C12 compounds detected (Figures 1 -10). PFOS was present !!l in the screened lots of commercial sera and plasma in concentrations ranging from 2.5 - 27 ng/mL and was present as branched and linear isomers. PFOA was present in the screened lots _<.! of commercial sera and plasma in concentrations ranging from 0.65 - 5.6 ng/mL and was present as branched and linear isomers. Accurate mass measurements and elemental compositions were obtained for endogenous C7C11, and PFOS that are consistent with theoretical mass and elemental composition. The presence of confirmatory daughter ions at characteristic retention times of target analytes in matrix are consistent with the linear isomer analytical standards. < i ..J ,t cJ Page 32 of 225 3M ENVIRONMENTAL LABORATORY E02-1039 REFERENCES i None LIST OF ATTACHMENTS AttachmentA: Extractionand AnalyticalMethods AttachmentB: Data Summary Tables AttachmentC: Sample Chromatograms , Attachment D: Sample Prep Sheets, Test Substance Information and Notes to File Attachment E: Protocol, Protocol Amendments and Deviations 't ':i REPORT REVISIONS I ':...1 Semi-quantitative estimates for THPFOS and THPFDS were removed from tables la, lb, 5a, and 5b because the data was intended as quantitative and only screening estimates could be :' obtained. References to THPFOS and THPFDS were removed from the list of test substances table 2 (page 11), chemical structures (page 13), reference substances table 3 (page 14), mass spectrometer acquisition parameters (page 18), and control of bias (page 19). References to THPFOS and THPFDS were also removed from the accurate mass determination discussion (page 20) and accurate mass determination data table 6 (page 21), daughter ion discussion (page 21) and daughter ion data table 7 (page 21), isomer discussion (page 21), and isomer figures 8 and 9 (pages 30 and 31). Wording in the Executive Summary (page 8), Summary (page ii_ 9re),mIonvtraoldoufctthioens(epmagi-equ1a1n)t,itaantidveSteastteimaetnetsofoCr TonHcPluFsOioSn a(pnadgTeH3P4F) DwSas. mSeovdeifrieadl tytoporegfrlaepcht itchael errors were also corrected, including William K. Reagen's phone number (page 7), formulas for the anionic forms of THPFOS and THPFDS in table 2 (page 11), and the units designator for %_ mass deviation, table 6 (page 21). A protocol amendment was written to address the removal of THPFOS and THPFDS from the study. I t_ Page 33 of 225 i _Li _i SIG NATU RE PAG E 3M ENVIRONMENTAL LABORATORY E02-1039 ,_ We certify that this report is a true and complete representation of the data for this study: M_k I_.Eliefson Study Director .... William K. Reagen Testing Facility Management Date Date _J I ,,t ;_:_. I Page 34 of 225 i ATTACHMENAT: METHOD :! i; i --a 5_al -y-- 3M ENVIRONMENTAL LABORATORY E02-1039 Page 35 of 225 3M ENVIRONMENTAL LABORATORY ,_ E02-1039 -_ .... ' . _,',.',>=_ i ' , _'-_;,i _] ":'7! Record of Deviation I. Identification Study/Project No. Deviation Tj_pe: (Checkone) E02-1039 r=]soP: .... I'-IProtocol ...................... _victhod E]EquipmentProcedt_e [3 Other: ................. Document Numl_eri i_'_S-8-23 I_i ....... . ..... Date(si of 0eeu_ene-- i0 C)et 02 to"2_i_Det-'02--- II. Description: Required Procedure/process: ETS-8-231.I section 9.6, "Quality Control (QC) Sample" see method for details. Ae_all'rocedure/proe_sssi_Tlecontinued accuracy of the calibration curve was monitored through the re-injection era curve point after no more than I0 samples and at the end of the run, not by preparing additional samples. The QC samples described in the method were prepared to monitor extraction efficiency. The QC samples were only spiked at 2 levels, 0.5 and 5ppb. In addition, only 2 QC samples were prepared for each matrix. It was possible, therefore, to distinguish between the continued accuracy of the calibration curve and the .extractio n effiy!en_ey.in eachma__.tested. _................................ DuHngthe course 0f flaestudy 2/3 of thi,-Qc sanapi-esprepm:ed in the sarne mah:ix-_-the ...... icalibration curve were within +9-5% of expected values, All of the 5ppb spikes passed, but only 3 of 9 low s_pikespassed. IlL Actions Taken: .............................. (s.uch_._.a..men d__m_e_nstued,_So_P..r.e vision,..et_c..:.)........................... No addit!onal..a.ctign wi!l.b_e.ne_ed__e.._..d............................................................. .............. ........ i? .........................................]7.....71......... i................ Date: Recorded By:, IV. Impact on Study/Project tt ThiSstudy.deviatdioorets n0t adversely affect the quality o-fthe data in the"context-o-f-the current .......... _'_._-_|t Authorized By: (Study Di_ior//Pro-j_-fiej(d) ........................... Date: .......-./ _ .......... Ll/l- I _ 3M Em,ironmental Laboratoly ,J " [ Deviation No. ( (assignedby StudyDirectororProjectLeadat the endofstudyor project) Page 36 of 225 _7 ,,L:5 i ,...... _'_ ii,i=-i:o 7_J :..-. 3M ENVIRONMENTAL LABORATORY E02-1039 .- Method Solid Phase Extraction and Analysis of Fluorochemical Compoundsfrom Biological Matrices Method Number: ETS.8.231.1 Adoption Date1: 1/13/_/ Revision Date: ._//_t/o2.. EffectiveDate.:_/#l/oz Approved By: Willliam K. Reagen Date ,4 $ ETS-8-231.1 Solid Phase Extraction and Analysis ot'Fluorochcmical Compoundsfrom Biological Matrices Page I of 19 Page 37 of 225 :Ui ':_!';! 7; _ i ; ! _,:;_I 2_._':_:i '"-:i ] i i,_J i_'I _:_._'._ _._';_,_ i i L':_.:.ii :i _;_i_ ii .... 3M ENVIRONMENTAL LABORATORY E02-1039 1 Scope and Application This =trothedescribestheextractionof ta._.'t analytasfrom fish,rat liver, ratscra,mouseliver, andmouseserausing solidphaseextraction(SPE). This methodmayalsobe extendedto otherbiologicalrnaciccsprovideddmtthe dam qualityobjectivesam met. 2 Method Summary An amount of biological ma),'rial,dctcnnmed by tic analyst.,is prepared(t_dds diluted and tissues homogenized) at a 1/6 dilution, or other dilution as dcte_ned by the analyst using reagent grade water. An aliquot of the dilution/homogcnatics spikedwith tic appropriatesurrogateor anal.ytm ixture. Acctonitzile(ACN) isaddedasan extractionsolvenatndalsoservetso p_cipitatcthe proteins.The sampleis capped,mixed'andputonthe cenn'ifice to Clarifythe supemata_._ supcrra,tam istransferredto a cleanrobe,dilutedv_thwater, andpassedtkrougha proeondifionedC,s SPE carCidgc.Finally, theu_aiytcsofintercs_amelatedfrom theSPE cartridgeandanalyzedby high pcrf.oml_cc liquid chromatography-clectzosptraanydemmassspccUometz(7HPLC-ES/MS/MS). 3 Definitions 3.1 Dilution A dilution expressedas 1:5or 1/6 is definedas: ! mL of.sample+ 5 m[J of.diluentfor a total of 6 mLs combined, unlessotherwls= noted. 3,2 SPE cartridge A columncontaininganopensolventreservoirat oneendandpackedwith bondedsilicasorbentsat the other end. It is designedto retain the compoundsof"inter_t under some solventconditionsand elut= them under others. A separation is thus achieved; compounds can b removed from difficult biological mamccs and introduced into appropriatesolvents for analysis. 3,3 Reagent grade water Water with no detectableconcentration(so)f the targetanalyt(s). 3.4 Quality control sample Sampleused re monitor_c cxwacdone_cicncy (-',_a matrixspike)andm _rif T rhccondnuedaccuracyof"_ minal calibration curve (as a continuingcalibration verification). 4 Warnings and Cautions 4.1 Health and Safety Warnings Always wear appropriate gloves, eyewear, and clothing when working with solvents, _mples and/or equipmenL Use caution with the voltage cables tbr the pr'ob. When engaged, the probeemploys a voltage of approximately 5000 volts. 4.2 Cautions Take cam not to allow the SPE column to run to dryness after the methanol and water washes. After washing is complete, add sample then allow all of the liquid to pass through the SPE column to dryness. ETS-8-23 I.I SolklPhase Extraction and Analysis of Fluorochemieal Conlpottndfrsom Biological Matrices Page2 of19 _ c_ '_ ___ J__ .,_ Page 38 of 225 I i:' i_:'_,'_ :'_'Ij I,!_ i_i,,:] !till] _::_ili ! ' '_: :i i_ : ' "7.-, 3M ENVIRONMENTAL LABORATORY E02-1039 Dbaor,ntehteoHpPerLaCtewsilol linvietinaptto-raauptosamboavtiecschauptadcoitvymof. 400 bar (5800 psi)back pressure, if the back pressure exce_is 400 Do notmn solventpumpsto dryness. 5 interferences To minimize interferencesT, eflon shouldnotbe ascdfor samplestorageor any partof instrumentationthatcomesin contactwith the san_[cor cxtracL 6 instrumentationS,uppliesa, ndMaterials The following instrumentation,supplies,and materialsarc used while performing this method. Equivalent instrumentations, upplies,andmaterialsmayI_ usedin placeof thoselisted. 6.1 Instrumentation Vortex mixer, VWR, Vortex Genie2 URra-TurraxT25 tissuehomogenizer va,,mP SPE ExtractionManifold Centrifuge,Mistral !000 or IEC Shakcr,EbcrbachorVWR Balance(+/- O.lO00g) Micromess, Quattro II or Ultima triplequadrepolc Mass Spectrometerequipped with an electro.sprayionization source HP l 100 or Agilent low pulse solvent pumping system, solvent degasser,column compartment, and autosampler 6.2 Supplies and Materials Eppendorf or disposable pipettes, plastic or _Jass Dissecting sealpcts Polypropylenc bottles, capable of holding 50 mL to I L (Nalgene) Volumetric flasks, glass, type A 40 mL glass vhals (/CH_ Plastic sampul vials, Wheatoth 6 mL (or other appropriate size) Centrifuge tubes, polypmpylene, 15 mL and 50 mL Labels GSyraridnugaetse,dcappipaebtlteeso, fgmlaesassuring 5 I.tLto 1000 I.tL Bottle-Top Dispenser (capable of'dispensing 5mL of solvent) SPE oxtmction cartridge, l g, Sep-Pak 6 cc tri-funetional C,, (Waters) 75 mL sample reservoir (or other appropriate size) Crimp cap glass autovials and caps ETS-8-23 I.I Solid Phase Extraction and Analysis of Fluorochemical Compounds from Biological Matrices Page 3 of I9 .._ "0_ _C_ _ >, ._ "g ,._ Page 39 of 225 : ,J !- I :1 :_ ....i. _:I ,_IL: '_ , i:_:! TI J:. _ ! .... i"_l, ] .' [!i_:__,._i ...... '..._i.', i,;! ".-. 3M ENVIRONMENTALLABORATORY E02-1039 Crimpers HPLCanalyticaclolumns, pecifictso bedeterminebdytheanalysatnddocumenteindtherawdata. 7 Reagents and Standards ReagengtradewaterM, illi-QTM,NanopumI1,orequivalent Acctonitr/lH, TLCgrac_orequivalent Methanol,H'PLCgradeor _[uivalent Ammoniumacetater,eagentgradeor equivalent Biologicaflluidsortissuesfr,ozenfromsupplier 7.1 Reagentspreparation 2.0 mM ammoniumacetatesolution:Weighapproximate0ly.300g ammoniumacetateP. ourintoa 2000 mL volumetricontainecrontaininrgeagengtradwater,mixuntilallsolidsaredissolvedb,ringto'volumeusing==agmt gradewater.Storeatroomtemperature.. Notet Whenpreparingdifferentvoluracsthanthoselistedinreagentspreparation,targetanal],,smtandardpreparation, andsurrogatsetandarpdrepacatioand,jusatccordingly. 7.2 Target analytestandardpreparation Prcparetargcat nalytestandard(sfo) rthestandardcurve.MulticomponeanntalytestandardasreacceptableT.he followingis anexamploenlyandmayormaynotbeappropriaftoerallstandarpdreparations. Weighapproximatelyt00 mg of targetan'ak_ intoa I00 mL volumetricflaskand recordthe actual weightin the aandardlogbookorotherappropriatleocation. Bringto volumewithmethanolfora stockstandardof approximately1000ppm(gg/mL). caIculat_onth.'eDilLsittoeck|000_._i.LSn'utin,w2(ithmL0/,0_thanlmL-'rwors_tai.n._gg,0/mL. s_Lrldard' slution of" approxilff3_tely ,0 ppilz F._J_amp I e Diluteworking standard1 withmethanolto producea workingstandard2 solutionof approx.5,0 ppra. Example cDail/cttumlawtioornk:5in0gIs.ttga/nmdaLxrd1I0mwiltJhl0m0emthLan=o5l.t0opp.rgo/dmuLcea workingstandard3 solutionof approx.0.50 pp_ Example calculation:50 lag/mLx 1.0mL/100aft.,= 0.5 I,tg/mL 7.3 SPrueprraoresguarrtoegsattsaetannddaarrdd(spT)r.heepfoallroawtiinoginsanexamploenlyandmayor maynotbeappmpdatfeorall surrogato standardpreparations. Weighapproximately90-110mgofsunogatestandardintoa 100-mLvohunetrieflaskand recordthe actualweight. Bringtovolumewithmethanolfora surrogatestandardstockof approximatel9y00 - I 100ppm. volumetricflaskand bringtovolumewithmethanolfora workingstandardof90-110ppm.Recordthe actual volume tral'isferred and standardconcentrationins the standardslogbookorotherappropriatelocation. Preparea sun'ogatestandardworkingstandard.TransferapproximatelyI mL of surrogatestandardstack toa 10-mI.. Prcparainternalstandard(s).The followingis an exampleonly andmay or maynot be appropriateforall internal 7.4 Istnatnedranrdaplrepstaarantidoanrs.d preparation Weighapproximately90-1I0 mgof internal:;tandardintoa 100-mLvolumetricflaskandrecordthe actualweight. ffrs-8-231.1 Solid Phase Exwactionand Analysisof Fluorochemieal Compounds fromBiologicalMatrices Page 4 of 19 _i ._ " _._ _@ _/ Page 40 of 225 ' .,, ':!_i ; ! ! _,_,_ i_i_it -! i_ ' i L i .... _ _:I ;_,_,, ',i,,J !,_ i _ ' ! :_ 'L.'" 3M ENVIRONMENTAL LABORATORY E02-1039 ".. Bring tovolumewith methanolfor an internasl tandardstockofapproximatel9y00 - I 100ppm. Preparean internalstandardworking standard.Tranffcr approximately1 mL of internalstandardstockto a 10-mL volumetricflask andbring to volumewith methanolfor a workingstandardofg0-110Fpnt Recordtheactualvolume transferredandstandardconcentratiomin thestandardsIogbeokor otherappropriatelocation. 8 Sample Handling i All smnplesam receivedfiozenandtram be keptfrozenmad[theextractionis performed. Allow samplestothaw toroomtemperatureprior toextraction. Typically fi-e_ matrix standardsarc preparedwith eachanalysis. Extractedstandardsand samplesarc storedin cappedautovialsuntil aral_is, / If analysiswill be dela)cd,_lracted stand='d.a_nd _amplesmaybe refiige_ted at approximately4C indefinitelyor may be storedat roomtemperatmuntil anal_iscanbeperformed. 9 Quality Control 9.1 Blanks 9.1.1 Solvent Blank An aliquotof methanoils usedasa solventblank. Solventblanksate notextracted. 9.1.2 Method Blank An aliquot of 1.0 mL of water, or other appropriateamount, is used as a method blank. Four method blanks am extracted and analyzed with each sut following this procedure (two are spiked with surrogate and two am not spiked), 9.1.3 Matrix Blank An aliquot of 1.0 mL or 1.0g of matrix(diluted orhomogenized) is used as a matrix blank. Other amounts may be used, as appropriate. Matrix blanks am l:_parcd fromone of throe source,s: l) a study control matrix from a study control animal received with a sampl set; 2) a commercially obtained sampl," of the same species as the study animals; or 3) a surrogate n-am'ix,also obtainedcommemiidly, but of a different species than the study animal. (g. if rat is used to gencrat standard eta'yen and CCVs for a mouse study). The matrix to use is dependent on the 9.1.3.1 matriSxtuusdeydcfoonrttrhoelmcuartvreix, curve- if the studycontrolmamxisusedfor the curve,preparefour(4) matrix blanks using the study control matrix(two spiked with surrogateand two not spiked). 99..11..33..23 curve, prepare four (4) zmlzix blanks using the same commercially available matrix (two spiked with surrogate and two not spiked). CSuormromgaetreciamllaytrioxbtcaui_rvde -(siafmaesusrpreocgiaetse) mmaattrriixxicsuu_see-d fiofrththee ccoumrvme,eprcmiaplalyr_ofbotuairn(e4d) m_mattrriixx ibslaunskeds ufosirntghethe same commercially available matrix and prepare four (4) matrix blanks using a commercially available matrix of 9.1.3,4 If limited matrix is available, the number of method and matrix blanks troy be adjusted and will he noted in tthheessatmudeysppreoctioecsoalsotrhienstthuedryaawnidmaatali.s(_.va spiked with surrogate and two not spiked). 9.2 Sample Replicate Samples replicates arc preparedaccording to each study protocol or project outline. ETS-8-231.1 Solid Phase Extraction and Analysis of Fluorochemical Compounds from Biological Matrices Page 5 of 19 "_= ._ ,'_.,, __ '_ 4 __' _/_ Page 41 of 225 3M ENVIRONMENTAL LABORA TORY E02-1039 L, _,,iI i i _;"_ I<''_ I , LJ _":} i_ ] !_<_ _:_, "_.i_ !'..:j _ :il 4i : _il : :; i,_J,:,_. _ ": i, ,! i 9.3 Surrogate standard If surrogatestandardis a componenot f thestudy,all samplesarespiked with surrogatestandardprior to extractionto obtain a concentrationin the mid-rangeof the calibrationcurve,with the cxccptionof blank sampleaas described above. Typically sumogatstandard_ spikedintotl_ 1.0mL diluted/homogenizesdampleremovedfor extraction.However, surrogate may be spiked din:ctly into the matrix prior to dihdng with water, into the diluted/homogenJznd sample priorto removing the 1.0 mL smnple, or into _ 1.0 mL di[uted&omogeniz=dsample removed for extraction. 9.4 Internal standard If internal standard is a component of thest-,_y, all samples arespiked with internalsnmdardafter extraction to obtain a concentration in the mid-hingeof thecalibration curve. Typically internal stand',a'idsspik_ into the 2.0 mL of exact in the 15 rnLcentrifuge tube, before transferringto the autovial. 9.5 Lab Control Sample Lab control samples arcnot a component of this method. 9.6 Quality Control lOG) Sample Preparequality control(QC) samplesto monitorextractionefficiencyand to vexi_ the continued accuracyof the initial calibration curve. Typically 1.0 mL, c.rother appropriateamount,ofthe same matrix used to prepare the initial calibrationcurveis usedforeachQC sampM. Twelve (I 2) quality controlsamples(QC) will be preparedforeachmatrixduringtbe courseof a study. A minimum of 3 QC samples must be prepared (one at e,',,'hlevel) on each day of sample extraction. (e.g. If the study is such that samples will be extracted on throedifferentdays then four QC samplesmust I_ prepared on each day of extraction for a total of twelw.) QC samples will consist of four samples at each of three levels of analyte` The levels listed below may be used and mLoawy rleevperle: s3eXnsttoam5XpltehceoLnLcOenQt,rationdsilutedintothe rangeof thecalibrationcurve: Mid-level: equivalentto a point neartic middleof thecalibrationcurve, High level: 80% of the ULOQ Two QC sample levels am analyzed aR=- every tenth sample injection starting after tim last calibration standard injection, with a minimum of three QC pea"analysis. Solvent blanksare not considered samples but may be included as such for determining wlmn QC samplns will Ix:analyz.cd. QC samples extra, ted with a particularsample set must be analyzed in the same analytical am. Any QC samplea extracted during thecourse of the study may he includedin subsequent analyses, If samples from multiple extraction dates are analyzed in one analytical ran, then QC samples from the same sample extraction dates must be included in that analysis. 'EachQC isexpectetdoshowanaccuracoyf75-125%ofexpactcdA. minimumoffalo3fallQC samplesmustmecr this criteria, and a minimum of I,'2of the QC samples at each level must mect this criteria. If not. the set must either bc re-analyzedor re-extracted. Any sample with an area greater than that of the highest acceptable standard will need to be diluted into the range of 9.7 StheacmalpiblraetioDn icluurtvieo. nIf samples are diluted into the range of the curve during analyses and enough sample remains, a post-run dilution validation will be peffonned to verify sample values. To perfbrm the dilution validation, one sample will be separated into two representative samples (i.e. two I.O mL aliquots for fluid samples or two 1.0 breamamounts for tissoe samples, or other amonnt as detcnnined by the analyst ETS-8-231. I Solid Phase Extraction and Analysis of Fluoroehemical Compounds fi'omBiological Matrices Page 6 of 19 _[ .t= "_" '_ ---_l _ _ ._. Page 42 of 225 3M ENVIRONMENTAL LABORATORY i E02-1039 I '_!'_:: _"Imq ,! i,_,:i _i:iii_ i_i!l ' ,_ ' .i I _!_ '_i;t"_i::i' -, anddocumentedina notetofilet)hendiluteudsingtwoproceduresT.he firsptrocedurceonsistosfdilutintghe samplewithadditionmaaltrixpriotro extracti(ofnluiadddingfluidw)h,ilethesecondprocedurceonsistosfdiluting thecx_actwithsolvenptost-cxa'ac(tmico_n.2noeixtracatddingadditionmaelthanoslolvcnt.J Iftherelatipvercendtifferenicsenotwithin15% forthesetwosamplesa;dditiontaelstinwgillbe requiretdo dctcnninewhichva[ucis acorrectrepre.sen_',ioonf thesampleconcentration. 10 Calibration and Standardlzatlon 10.1 InstrumentCalibration Onecalibrationcm-vcwill be preparedfromextractedmatrixstandardsi,n tim samematrix asthe samplesp, er study. Itwillcomistofa minimum ofnino(9)l_cls.Additioncaallibratoiuornvcsmay beexaacteodn scpamm sample extraction dates, as determinedby the analystand documented in a note to file. Transfer 1.0 mL, or other appropriateamount, of diluted control fluid or homogenized control tissue to a 15 mL centrifugetube usinga disposableplasticpipette. This will be repeatedwhilepreparingaliquotsfor the standard creumrovveed. .Be sum to mix or shakethe controlmatrix containerbetweenaliquotsto ensurea homogenoussample is RecordeachstandarvdolumeontlmwcightA,oluns,h_eeotrextraction wo_ as appropriam. Four 1.0 mL aliquots, or other appropriateamount, of control matrix serveas matrixblanks. Tho standarcdoncentratiaondsspikinagmountslisteind Tablel may be used,when appropriateo,spikeone standarcdurve.A totaolf9 standardfso,urmatribxlanksa,ndfourmethodblankas_ prepareidnadditiotnotheQC samplcaand testsamples.The numberofstandardasndblanksmay be adjusteadsdeterminebdy theanalysatnd documentedina notetofile. Use A:tachmcnt C, or other appropriatefonn, as an aid in calculating tic concentrations of the working standards. Refer to section 12 to calculate the actual concentration ofanalyto in each calibration standard and QC sample. Typically the target nnnlytestandardis spi_.'d into the I.OmL d/luted/homogenizedsampleremoved for extraction. However, it may he spikeddirectlyinto the matrixprior to diluting with water,into the diluted/homogenizesdample. prior toremovingthe l.O mL sm'npleo,r intothe 1.0mL diluted/homogenizesdamplerr.n_vcd forextraction. Analyzethe extractedmatrix standardcurvepriorto eachsctof extracts.The curveequationwill be determinedby regression analysis using the peak areasof the target analyte(s) using MassLyax or other suitable soRwar. Any level outside 75% - 125%of nominal lr_St be deactivated, and regression rc.-calculatcd, except the LLOO which must b within 3_A ofnominaL All levels must show a response greater than twice that of the blank. A n-aximum of thr_e (3) levels may be deactivated in any one set, or the set wilt be re-analyzed. --f+ _ ii ETS-8-23 L ! Page 7 of]9 , Solid Phaso Extr,zction and Analysis of Fluorochcmica! + Compounds from Biological Matrices Page 43 of 225 i>__l ;:i_ i :, ,:,', .... !i_j 'i : _,>:,i r:,; t_,il ....: ii i { 3M ENVIRONMENTAL LABORATORY E02-1039 "-" Working standard (approximate concentration) 0.500 u_)mL 0.500u_/mL 0.500u[/mL 0.500 u[/mL 0.500 u_/mL . 5.00u[,'mL 5.00u_mL 5.00u_mL 5.00u[/mL 5.00u_/mL 550..000uus_//mmLL 50.0 u_/mL Surrogate Std 100 ug/mL 11 Procedures Approximate final cionncMenattrriaxtiondiluofteadna1l:y5t Blank 1.5 5.00 ng/g or ng/mL 3.0 I0.0ng/gorn[/mL 8.0 25.0ng/gorn_/mL 16 50.0 ng/g or ng/mL 32 100 ng/g or ng/mL 5.6 175ng/gorns/mL 8.0 250 ng/gorng/mL 16 500 n_/_orn_mL 24 750 n_/gorn_/mL 32 I000n_/gorn[/mL 54.00 11520500nn[_]/_,oorrnng[//mmLL 6.0 1750 n_ or n[/mL I0 6500 ng/g or ng/mL Approximate final Blank 0.375 ng/mL 0.750ns/mL 2.00n_/mL 4.00 ng/mL 8.00 ng/mL 14.0n_mL 20.0n_mL 40.0n_/mL 60.0n_/mL 80.0n_/mL 112050nn[_/mmLL 150 n_/mL I 500 ng/mL 11.1 TissueSamplePreparation Obtain fi-ozentissue smnples Cut approximately 1.0000 g of tissue (+/- 0.1000 g), or other appropriate amount, using a dissecting scalpel. of the procedure is bestperformed quickly, not allowing the tissue to thaw. Weigh the tissue directly into atared plastic sampuin vial. This part Record the weight on the weight/volume shccl, extraction werksheet, or other appropriate location. Return unused tissue to the freezer afierextraction amounts have been removed. Add 2.5 mL of reagent water to sampule vial, or other voJume as determined by the analyst and documented in a note to file. Homogenize the sample. Put the Ultra-Tumm grinder probe in the sample and grind for approximately 2 minutes, or until the sample is homogeneous. Rinse the probe into the tube containing the sample with 2.5 mL of reagentgrade water, or other volun_ as determined by the analyst and documented in a note m file, using a pipette. Take the grinder apart and clean it with metlumoi alter each sampte. Refer to ET5-9-52 for more information. If an amount other than 1.0000 g (not within +/- 0.1000 g) is removed for an initial weight, adjust the water volume" ratio as determined by the analyst and documented in a note to file. _ %| __ O '_ ETS-8-231.1 Solid Phase Extraction and Analysis of Fluoroehemical Compounds from Biological Matrices Page 8 of 19 Page 44 of 225 3M ENVIRONMENTAL I.ABORA TORY : E02-1039 'I _'t :i : ."'_ i _, , :i :_ _ _-(:_ _':_ ....._. ::_j !i_._;_ 11.2 Fluid Sample Preparation Obtainfi'ozcnfluidsampleandallowit tothawat roomtemperatureor in lukewarmwater. Label a 15 mL polypmpylenece.ntrifogctube with the studynumbers, ampleID, extractiondateandanalystinitials. Seeattachedworkshcct(AttachmentA or similarworkshect)for documentingtheremainingsteps. Vortex mix thefluid sampleforapproximately15 secondsth, entransfer1.0mL of fluid,or otherappropriateamount to a plasticsampulcvial,or otherappropriatecontainer. Returnunusedsamplesto fi'ccz=ar .Rcrextractionamountshavebeenremoved. Add 5.0 mL of reagentwaterto the 1.0mL of tluidfor a 1/6 dilution,or otherdilutionas determinedby the analyst anddocumentedin a notetofile. If a volume other than l.O mL is removed for an initialvolume, adjustthe water volume accordingly to maintain the same dilution as above. 11.3 Tissue and FluidSample Extraction ARer tissue or fluid samples have bccn prt:pamd according to sections l 1.1 and 11.2, vortex mix or sMke by hand the diluted/homogenized sample for approximately [5 seconds then transfer 1.0 mL, or other appropriate volume, to a clean ! 5 mL polypmpylcne.cntrifugetube. Rcmmunuseddilumd/homogcnizcdportionsto the freezeraRe.rextractionamountshavebccnremoved. Recordthe volumeremovedonthe extractionworksheet(,AttachmentA orsimilar worksheet). Spikeblankss,amplesa,ndstandardrse,adytbr extractiwointhsurrogastteandaradsdescribeidnthimsethod. Spike each calibration standard marx with the appropriate amount of standard as described in this method for the calibration curve standardsand c'achQC sample. Vortex mix the standard curve samples and QC samplasfor approxilmtcly 5 seconds. To cachsmnpleandsmndsrda,dd5.0mL ofacctonitrcialpc,,andvortexmixorshakeby handappmximazel1y5 seconds. Place all samples on the shaker at anappmpr/ate speed for 20 minutes to adequately mix (a setting of approximately 300 rpmon themodelslistcdinsection6.1). Rcmov_ from the shakerand centrifogeat anappropriatespeedfor 10 minutesto adequatelypellet the precipitate(a setting of approximately 2000 rpm onthe models listcd in section 6.1). Add 40.0mL ofreagengtradewatertoa clean50 mL centrifutguebe.Remove samplesfromthecentrifuagned decantthesupernatainnttotimwaterinth,5-0 mL tubet,akincgam nottointxoduacneyofthematrixsolidisntothe solutionC.ap andmix by inwrthlsgcvratliffinIsn.thistopelmonlcrofadditiomnay bcctmngcd(i.et.imsarnpt AnxttaaycbhethpeutriensteorvthoiercteontthriefuSgPeEtucbaertzainddgethanend tahtetawcahtethriasdredseedr)v. oir/cartridgeunit to a vacuum manifold. flow of 5-7 mL/min. Flows ,nay vary thmul;h cartridgcsand the kPA =naybe raised for slow tubes and deying aRcr most hav beendrown down. NPr_cTpEar: Wtbeh_SnPEmncnarintrgidthgee _baynwuaushmins_g_tttwh_icveawciuthumacplpmromxbimcraatetlayp5_.0r_mxiLmoaftem_ey_t_haknPoAl, -fot_llogwive_dabnya_apppmrxoixmimaarte_luytti_wo Aflcr washing is complete, pour the samph: into tim reservoir/cartridgeunit and allow all of the liquid to pass 5th.0romugLhaltiqhueoctosluomf wn ateord, rtyankeinssg. care not to :allow the column to run to dryness after each wash. Run die vacuum on high for approximately 5 minutes to adequatelycityeach SPE cartridge. Place a collection 15 mL polypropylcn-"centfifiagtube under each cartridgeand eiute with 2.0 mL of methanol Spike cxtractcd blanks,samplcsa, nd standardswith internal standard as dcscribcdin this method. ETS-8-23 I.I Solid Phase Extraction and Analysis of Fluorochcmical Compoundsfrom Biological Matrices Page 9 of 19 .'_ o "o_-.'__ ___ r_e" _'/'--_ Page 45 of 225 : ,_ _.4 ' "_ _i:":_ ;,:_,1 ii..... ["1 :I _ _ _,,, _!,t_ !_" 3M ENVIRONMENTAL LABORATORY E02-1039 Labol each glass autovial, as appropriate, with the study number, vial rite archive number, animal numbcr/gendcdtiroepoint or LIMS number, matrix, final solvent, analyt components (if needed), extraction type, extraction date, and analyst(s)pcrfmming the extraction. Transfer each oluant to a glassautovial and cap. 11.4 Extract Analysis 11.4.1 Software set-up On the MassLynx main page, set up a sample list name. Save the list as instrumentdesignatorletter, last 2 digits of test year-month-day, and a letterthat will increase throughthe alphabet with each additionallist for thatday. F..xamplcSample List:FfYMMDDa or A020204a 1= Initial of the insmuncntname (A: "Amelia") YY = Testyear(02) MM ,, Testmonth(02) DD-,o da,O,) a ",F'irstsample list (run)ofthe day (thenext sample list will end with _', the next 'c', and so on.) Assign a filenanm using the instrumentdesignator letter, the last 2 digits of the tc_ year-month-day, and a 3-digit sequential file numberthatstartswith I and iacma_s by one foreach filenamc. Example filcname: IYYMMDD/fl#?or A020204001 I = Initial ofinstmmcm name YY = Test year MM - Test month DD: Test day ### : 3-digit sequential file numberstartingwith I through999 (001) Also, as part of the samplclist,assign a method (MS) for acquiring, an inlet file, a bottle number,an injection volume, and sample dcscfiptiot_. To create a mothod, click on Method Editor button in the MS Status Pane and select SIR (Single Ion Recording) or MRM (Multiple Reaction Monitoring). Set Ionization Mode as appropriateand mass to 499 or other appropriate mass(s). Also set the acquisition start mad stop times. Save acquisition method. If MS/MS insm.zmentaax_ employed, additional product ion fragmentationinormadon may bc collected. See Microm,x_MassLynx "Guide to Typically the analytical batchrun I:cgins with system suitability,so|vent blanks, and a set of extractedmatrix standards. Data Acquisition," for additional iafotmatiOnoseuqMueRmMcc. Sample extracts are analyzed wifl_two QC samples injectedafter every tenth sample injection. Solvent blanks should. be analyzed periodically to monitor possibl," analyte carryover and am not considered sample extracts but may be included as such. --_ ._'l [ '_ "_ :L_'X ._ r"_ ETS-8-23 [. I Solid PhascExtraction and Analysis of FluorochcmicaJ Compotmds from Biological Matrices Page 10 of [9 Page 46 of 225 i ...... !_ :,i:JI i , ;x_ !: i I': T _il _._, !4 /!i ,_:,i _ :i : 3M ENVIRONMENTAL LABORATORY . E02-1039 11.4.2 HPLC set-up Set up sampletray accordingto the samplelistprepar_l above. Sct up theHPLC othe followingconditionsor at conditionstheanalystconsiderasppropriateforoptim._lresponse. Recordactualconditionsin d' immanentlogbook,orotherappropriatelocation: Samplesize= !0 HL injection Inject/sample= I Cycle ti,ne = 10.0 minutes Flow rate = 300 g/./min Mobilephase:SolvenAt ==2 mM Ammonium AcetateS,olvenBt=,Methanol Solventgradientprogram: Time % Solvent B 5.5O 95% 7.50" 8.00' 10%" 11.4.3Instrumenset t-up Rogerto ETS-9-24, "Operation and Mainteaance of the Micromass QuattroII Triple Quadrupole Mass Spectrometer Fitted with anAtmospheric PressurIoenization Source.fo,r"details. Checkthesolvent level in HPLC resewoirsand refill if necessary. Check the stainless steel capillaryat the end ofthe probe. Use aneyepiece to check d_etip. The tip should be fiat with no jagged edges. If the tip is found to be unsatisfactory,disassemblethe probe and replacethe stainlass steel capillary. Turnon tlcnitrogen. Open the mr,page. Click on operate to initiate source blockand d_olvation heaters. Open th_ Inlet l_dJtor. SDeotwthneloflaodwthteo/-I1r0p-L-5c0m0ueUthmodinaonrdaisnaidpaptreosporlivaetent flow to begin s3,stemequilibrium. SetI-[PLpCump to "On" Observderopleotrsmis_comingoutofthetipoftheprobe.A finmeistshouldbe expelled with no nitrog_ leaking around thedp of theprobe. Readjust g/ _ -4 ._ _ "x_ ,,.a ._ t Allow to equilitbhreattiepfoofrthaeppproroxibne_itfenlyo mI0ismt iisnuotbess,erved Typical instrument panuncters include: Drying gas 250-400 liters/hour ES nebulizing gas IO-15 litcrs/hour u_._ _;_ ETS-8-231I. Solid Phase Extraction and Analysis of Flnorochemical Compounds from Biological Matrices Page II of 19 Page 47 of 225 /:,l !i g :_::::i _,,_._ -! i _J' ..... i !i '1 _::'_ _!i _,:._I:.._] ,=::. i i. _ !_!:.:! :i 3M ENVIRONMENTAL LABORATORY E02-1039 HPLC constanftlow mode,flow rate10-500I.Z[Jmin Pressure<400 bar(thisparameteris notset,it isa guidetoensurethe HPLC is operatingcon_ctly.) Sourceblocktcmpcratureapproxin_tcly150"C Desolvation temperatureapproximately250C Thes,o set, rigsmay change in orderto optimize theresponse Print the tune page, sample list, and acquisition method from MassLynx and store it in the study binder with a copy taped into the instrument log. Click on start button in the Acquisition Control Panel (the location of the start button may vary among MassLynx versions, rcfcr to appropriateMossI.,ynxUser's Guide). t2 Data Analysis and Calculations 12.1 Calculations If othercalculationsareusedthanthoselisted,theywill be documentedinthe rawdata. Calculate the matrix amount containedin thc initial dilution using the following equation: Matrix Amount (g/mL or mL/mL) = IW (g) (or IV (mL)) (IW(g)(orIV(mL))+DV (mL) Calculataectuaclonccnu'atioofnasnalytiennalibmfisotnandarudssingth_followienqguation: Conccnrcation (ng/g or ng/mL) = Spike Concentration (ug/mL)x Spikcd.Amount(mL)xlO00 ng SV (mL)x Matrix Amount (g/mL or mL/mL) Iug IW = Initial weight (whom 1.0 g = 1.0 mL ) IV = Initial volurns DV _ Diluent volume (rw..agcnt grade wamr) AR = Analytical resultfrom MassLynx maunmry SDVF =,- DSialumtipolnevfoalcutomreremovedforcxtracdon(typlcally1.0mL) FV = Final volume MA ='Matrixamount 3 curve= MA oftissuc/fsltuainddarcdurvea,ssumedtobe lg orImL/5mL water 3 sample = MA of tissuc/fluid sampleC....,).r.m.Lg of sampld5 mL water) Calculate spike percent recoveries using the tbllowing equation: 'Y,,Recovery ObservedResult - Matrix Blank Result = Spiking Level x tO0 ETS-8-231.1 Solid Phase Extraction and Analysis of Fluorochcmical Compmmds from Biological Matrices Page 12 of 19 _ o's ''__----_m r2_o _ ! "%c__- _t'._,,,.3 Page 48 of 225 :i I ':' 'i '7_ ,_ ! i-_.! .J :_ i_ _!i_ :_:_"__:_:'i_'_'_! 'i i i L= = :,L; .!/_! 3M ENVIRONMENTAL LABORATORY E02-1039 Calculate relative standard deviation using the following equation: Standard Deviation RcladveStandard Deviation = x 100 Mean Calculate portent deviation using the foUowiag equation: %Deviation = Expected Cone.- Calculated Cone. x 100 ExpectedCone. Calculate actual concenmafionofanaJytein _uid (gg/mL): AR.(ng/mL) x DF x _ curve (mUm[.) x FV fmL) in Curve x 1.0 leg o3sample (mUm/..) FV (mL) in Maaix 1000 ng = (lag/g) Calculam actual conccotration ofanalytin tis,_ _g/g): AR (ng/g)x DF x 3 curve(E/mL_x FV (mL'_in Cuw x _ asample(g/mL) FV(mL)inMatrix 1000rig = _g/g) 13 MethodPerformance 13.1 SystemSuitability System suitability will be determined prior to the start and at the completion of each analytical run. Prior to the calibration curve and aRer the last sample of the run throe (.3) mid-lcv=l unextractcd calibration standards will be" a,mlyzcd. As applicable, the peak area precision, mtendon time pt_-cision,resolution, and peak asymmetry will be monitored at thc beginning and the end of the mn separately. The peak area precision must b equal to or less than 5.0% RSD, the precision of the retention tin_ must be equal to or l_s than 2.5% RSD, the resolution must be > 2.0, and the peak asTmmctry (fconting or tailing) must be 0.5<AF<2.0, where AF is the a_. nmetry factor. sIyfsatenmy istuemitaboiflittyhesasmysptleems asnuditatbheilistyystfeamilss,usiytastbeimlitymmauinsttepnaasnscbeefmoureststbaertcionmg tpbleetceadlibprraiotironto. mIf'nsynsitnegmasuseitcaobnidlityseftilolsf at the completion of a run,the sample set must be manalyzcd. The coefficient of determination value for the calibration curve, plotted by regressionusing the peak areas of the analyt_s), must be 0.990 or better. All active calibration curve points must bwithin 25% of the theoretical value with the exception of the LOQ point, which may deviate up to 30%. Calibration standards with peak areas less than two times the curve lm_x bhnk will be deactivated to disqualify' a data range that may be affected by backgroundlevels of tl_ analyt, A valid calibration curve must contain at least 6 active points above and includingthe LOQ. If the curvecannotmeetthesecrkcr/a,thesamplesetmustbc reanalyzcdor rccxtractcd. 13.3 Accuracy TWOthirds ofall quality control samples and }/2 ofcach quality eonWol sample at each level amexpcctcd m show an accuracy of 75-125%. ETS-8-231.I Solid Phase Extraction and Analysis of Fluorochcmical Compounds from Biological Matriccs Page 13 of 19 ""c_c:) '>_, ca. j _ " :_ u._ _,_'_ Page 49 of 225 'i 3M ENVIRONMENTAL LABORATORY ;i E02-1039 !,_)f .... _! Surrogates and internal standards must have a percent deviation < 50%. Deviations outside this range will be _i!J reanalyzed re confirm. If the second analysi;_conarms the original, the dcvistion wifl be documented in the raw data. Ifthc second analysis is within 50%,then the second value will replace the original value. ! 14 Pollution Prevention and Waste Management Samplewasteisdisposedof in noninFcc_oubsiohazardwastecontainers. Flammablesolventwasteisdisposedof in highBTU' containers. Glass pipette waste is disposed of in broken glass containers located in tl_ laborato_. ;,,:_ 15 Records ,.) j Completethe extractionwork_hectattachedtothismethod,or otherapplicableworkslcct andstorewith thestudyraw -_ data. ; " J Each page generated for a study mu.contain the following information (if applieabl,.): study/project or instrmnent number, acquisition method, integration method, sample ntur, extraction date, dilution factor (if applicable), and analyst. Other information may be added if applicable to the study. ' Print the tune page, sample list, and acquisition method from MassLynx to include with the study raw data. Copy __j these pages and tape into the instrument rtmlog. Plot the calibration curve by the appropriateregression. Print these graphs and store with the study raw data. Print data integration stmamary,integration method, and chromatograms from MassLynx, and store with the study raw data. Summarize data using suitable so.cam (Excel 7.0 or LIMS) and store in the study folder. Back up electronic data to appropriate medium. Record in study notebook the tile nanm and location of backup _!! electronic data. 16 Attachments _z_, Attachment A: -ExtractionWorkshcet Attatchrlncnt B; Sample We[_tFv'o|urlrze WOlkSheet Attachment C, Calibration Standard Concentration Worksheet -- _-i 17 References .... ETS-9-24, "Operation and Maintenance of the Micromass Quattro lI Triple Quadrupole Mass Spectrometer Fitted '_ with an Atmosphere Pressure Ionization Source" u._ ETS-9-52, "Operation and Maintenance of a Tissue Grinder" 18 Affected Documents '_ ETS-8-231.1 Page 14of19 i Solid Phase Extraction and Analysis of Fluorochemical Compounds from Biological Matrices Page 50 of 225 r!,,,_" 4_:i '11:13 _:_t I ' : 1 ! 3M ENVIRONMENTAL LABORATORY E02-1039 .... 19 Revisions R_visJon Numbcr I RevisionDescHnfion MinorformattingchangesA.ddeddetailedinformatiotno all secdoncsoncernintghe extractionprocedurea,nalyticaplrocedurea,ndcalculatioN.Addedattachmenatsnd references. Revision .D.ate 02/18/02 r _'_. h_ '! ETS-8-23I.I Page [5 of 19 Solid PhaseExtractionand Analysis of Fluoroche,nical Compoundsfrom BiologicalMatrices Page 5I of 225 ?!i "_i_:_l .!_-_7 i_/:-,_ --'1 i-{ ;i J f'; _i 3M ENVIRONMENTAL LABORATORY E02-1039 Study Number: Prep Date: Analysts initials: Box#: Attachment A - Extraction Worksheet Method Revision:ETS-8-231.1 Matrix: Sample Timepoint: SampleNumber Sample Number Volumeof Amounat nd or dilutedsample Amounatnd surrogatsepike Typeofcolumn Elutlonsolvent description removed spikemixused mixused usedandlot andvolume Comments .... .o, ....i :7 i i , _:__i '_':;1 ,i -i Blankmatrix TN-A- ;Amounwt eighedlaliquoted: g/mL 21.. HAoliqmuoog1temnLizsoeafdmilpulteedmatrixinto15mL polypropylenttd,be 3. Spike samples accordingly 4. Add N-- mL of ACN (TN-A- ) to each diluted sample and shake or vortex mix 5. Shakesamplefor20 min@ rpm(Shaker ) 6. Centrifugseamplefor10rain ___ rpm(Centrifuge1. ) 7. Add40 mLof -- waterto50 mLpolypmpdencaentrifugteube. 8. Decantextractinlocentrifugteubeswithwater 9. Shakesampleslicjhttytoensurepropemr ixing 1I. Conditioncolumnwithtwowashesof-5 mL MeOH(TN..A- ) - donotallowcolumntogotodP/ness 12. Washcolumnwithb.,vwoashesof-5 mL__ water- donotallowcolumnto goto dryness 1130.. AFitlttaecrsha6mmpLleCt1h8roSuPgEhccoanrdtriitdiogneesacdnodl7u5mmndL, irsecsaerdrvinofgiirltstoravtaecuummanifold 14. Allowcolumntogotodryness.Afterdrippingstopsd, rawa highvacuumthroughcolumnforat least5 minutes. 15. Elutecolumnwithsolvent( TN-A-___) intoappropriat1e5mLcentrifugteube 16. Spikesampleswith-- uLofinternasl tandard# 17. Transfersampleintoappropriatelyabeledautoviaal ndcap __ conc. Note: Invacuumstepsabovesetthevacuumchamberat ar_proximate1l5ykPA-thisshouldgiveapproximate5ly-7mL/mlnelutlonflow Flowsmayvarythroughcart,ridge-skPAmaybe raisedfor,';lowtubesanddPjingaftermosthavebeendrawndownandshutoff. ETS-8-231.1 Solid Phase Extraction and Analysis of Fh.torochemical Compounds from Biological Matrices Page 16of19 | __ O"_- ,_ 'J "_ oc_ _" J Page 52 of 225 3M ENVIRONMENTAL LABORA TORY E02-?039 r,: i ." i i Attachment B - Sample Weight/Volume Worksheet Prep Date(s): _! Analyst(s): Sample Matrix: Method/Revision: Study Number: Equipment Number: Final Solvent & TN Number: i Sample ID Initial WtdVnL Water Vnlume added Vnlume Removed _nmments _mlLA'f.., (mL) (mL) ,{ .... i .; Form Completion Verified By:. ETS-8-231.1 Solid Phase Extraction and Analysis of Fluorochemieal Compoands from Biological Matrices Pag* 17 of 19 Page 53 of 225 3M ENVIRONMENTAL LABORA TORY E02-1039 _ __ _.i _i_,_ i , _t.,J _] ! '_ _- J : :, _":_' , _1 'i , Prep date(s): Analyte(s): Sample matrix: Method/revision: AttachmentC: CalibrationStandardConcentrationWork,sheet Standard number: Equipment number: Final solvent and TN: Blank Tissue or Fluid/identifier: AnalySe mix std approx. 0.500 ng/mL: AnalySe mix std approx. 5.00 ug/mL: AnalySe mix std approx. 50.0 ug/mL: Surrogate std approx. 100 ug/mL: Actu tl concentrations of standards in the analyremix AnalySe Std cone ug/mL 0.500 0.500 0.500 0.500 0.500 5.00 5.00 5.00 5.00 5.00 5.01} 50.0 All Am't spiked mL 0.0015 0.0030 0.0080 0.0160 0.0320 0.0056 0.0080 0.0160 0.0240 0.0320 0.0400 0.0(15 All Final Volume: lr_ 2.00 2.00 2.00 2.00 2.00 2.00 2.00 2.00 ZOO 2.00 2.00 2.00 All Initial Fluid Dilution mL/mL 0.1667 0.1667 0.1667 0.1667 0.1667 0.1667 0.1667 0.1667 0.1667 0.1667 O.1667 O.1667 All Initial Tissue Density g/mL 0.1600 0.1600 0.1600 0.1600 0.1600 0.1600 0.1600 0.1600 0.1600 0.1600 O.1600 0.1600 50.0 0.006 2.00 O.1667 O.1600 Calculated concentrations of standards in relation to the final 2.0 mL solvent and initial matrix 2.0 mL Final Volume Fluid Matrix Tissue Matrix Analyse Final cone. ng/mL 0.375 Surrogate Std cone ng/mL 1O0 AnalySe Final cone. ng/,nL 5.00 Surrogate Std cone ng/mL 100 AnalySe Final cone. ng& 5.00 Surrogate Std cone ng/mL 100 O7. 50 2.00 4.00 Surrogate Final cone IO.O 25.0 50.0 Surrogate Final oanc I O.O 25.0 50.0 Surrogate Final cane 8.00 n_'mL 14.0 0.500 100 ng/mL I00 175 6500 175 n_l _ 6500 20.0 250 250 _ 4o.0 500 500 60.0. 80.0 , I O0 125 150 750 I000 1250 1500 1750 750 I000 1250 1500 1750 ETS-8-231.1 Solid Phase Extraction and Analysis of Fluorochcmical Compot,nds from Biological Matrices Page 18 of 19 ] "_ = _I ,.r:.:-, ._, >, t._ _ _ _ "_ _ _, Page 54 of 225 q _,_,,_ ii__i,'!! 'i 3M ENVIRONMENTALLABORATORY E02-1039 Study: DBioluxtNiounmDbaetre:/Analyst: Sample Number or Description 1/ Attachment D: Dilutions Summary Works, heet SolventfrN Number: MExattrraixct/TioinmDeaptoei/nAt:nalyst: 1/ I/ 11 1/ Dilutions 1/ 1/ Comments Verified By: I__,, N1/o1t0esd: ilution = of sample + of solvent _ __ ___ ri , :, Form Completion Verified By: ETS-8-231.1 Page 19 of 19 SoLidPhaseExtr,xctionand Analysisof Fluorochemical i CompoundsfromBiological Matrices Page 55 of 225 .. ii':i,i ATTACHMENT B: DATATABLES _7 3M ENVIRONMENTAL LABORATORY E02-1039 t :,7"1 :r Page 56 of 225 Human Serum Data Quantitated with Chinese Plasma Calibration Curve Alldataisin unitsn_/mL Lampire Serum PFOS sample 1 < LOQ sample 2 < LOQ low spike < LOQ IISigma Serum High spike] sample 1 sample 2 . low spike 5.47 4.74 4.38 4.75 Ct= < LOQ < LOQ 0.65 5.70 < LOQ < LOQ 0.45 CIt < LOQ < LOQ 0.82 7.02 < LOQ < LOQ 0.69 Ct0 < LOQ < LOQ 0.68 6.34 < LOQ < LOQ 0.60 C9 < LOQ < LOQ 0.79 5.91 0.19 0.34 0.84 C= 0.73 0.57 1.39 6.81 1.60 1.59 1.86 _z < LOQ < LOQ 0.75 6.51 < LOQ 0.15 0.65 High spike 9.06 3.74 5.45 5.17 5.91 6.64 5.78 lGolden West Serum I ] sample I sample 2 25.64 28.41 < LOQ < LOQ 0.31 0.33 < LOQ < LOQ 0.96 0.84 5.54 5.65 0.23 0.15 low spike 24.16 0.46 0.85 0.60 1.30 4.64 0.41 High spike 26.10 3.65 6.42 5.20 6.62 9.12 4.91 Biorasource Serum sample 1 16.79 < LOQ 0.28 < LOQ 0.63 2.83 < LOQ sample 2 17.26 < LOQ 0.31 < LOQ 0.58 3.06 < LOQ High spike 18.78 2.65 4.94 4.32 4.95 7.39 1.47 Analyte ... PFos Clz C, C1= C9 Cs C7 Calibration Range, nglmL 1.01 - 30.2 0.104 - 31.1 0.103 - 30.9 0.252 - 30.3 0.101 - 30.4 0.258 - 31.0 0.102 - 30.7 Ch!neese Plasma_ Serum Data Cal Curves _ystem Suitability Samples, CCV #_ CCV #2 CCV #3 R= 0.999596 %RSD 2.48 recovery, % recovery, % recovery, %= 52% 3% "" 0.999621 0.999767 0.999471 0.9997 0.998816 0.999569 0.99 2.46 1.45 2.94 6.73 2.37 77% 105% 86% 91% 94% 87% 52% 105% 90% 105% 87% 95% *" 99% "* 102% ** 93% Matrix Blanks < LOQ < LOQ < LOQ < LOQ < LOQ < LOCI < LOQ "None performed Low spike 16.44 < LOQ 0.96 0.73 1.11 4.03 0.18 0 t_ Human Plasma Data Quantitated with Chinese Plasma Calibration Curve All datais in unitsng/rnL Chinese Plasma PFO5 sample I < LOQ sample 2 < LOQ low spike 1.30 High spik 4.78 IILampire Plasma _ sample I sample 2 9.89 12.34 Clz < LOQ < LOQ 0.52 5.02 < LOQ < LOQ Ctl < LOQ < LOQ 0.65 5.18 < LOQ < LOQ C10 < LOQ < LOQ 0,40 4.88 < LOQ < LOQ C9 < LOQ < LOQ 0.59 4.91 0.37 0.50 low spike 13.15 0.48 0.46 0.49 0.78 Ca7 << LLOOQQ << LLOOQQ <0.L2O8 Q 45..7218 20..3200 <2.L9O1 Q 30..3542 High spike 16.19 4.53 4.42 4.01 4.88 75.,3413 Golden West Plasma sample I 16.20 < LOQ < LOQ 0.15 0.42 sample 2 20.32 < LOQ 0.10 0.17 0.65 low spike 13.56 0.44 0.52 0.57 0.72 03..4456 40..2183 30..3342 High spike 20.34 4.80 4.95 5.29 5.11 Ilnnovative I _ sample I 15.61 < LOQ 0.13 0.19 0.68 Research Ptasma sample 2 15.55 < LOQ 0.14 0.15 0.49 low spike 17.32 0.58 0.84 0.78 0.84 75..9344 <3.L0O4 Q <3.L0O9 Q 30..8468 High spike 21.63 5.66 6.16 6.08 6.00 96..1164 Analyte PFOS C_z Cll Cl0 C9 .Cs ICT Calibration Range, nglmL 1.0 - 30.0 0.10 - 31.0 0.10 - 30.9 0.10 - 30.3 0.25 - 30.4 0.52 - 31.0 0.10 - 30.7 Chineese Plasma i Plasma Data Cal Curves _ystem Suitability Samples, CCV #1 CCV #2 Matrix R2 0.999721 %RSD 2.20 recovery, % recovery, % 102% 84% Blanks < LOQ 0.999759 0.999585 5,87 1.21 101% 90% 84% 78% < LOQ < LOQ 0.999695 2.76 0.999847 5.07 91% 97% 83% 80% < LOQ < LOQ 0.999736 0.999695 2.85 5.85 81% 102% 89% 75% < LOQ < LOQ '-o O0 0 Human Serum Data, 5x Sample Scale-Upwith 20uL Injection Volume Quantitated with Chinese Plasma CalibrationCurve All data is in units ng/rnL Lampire Serum Sigma Serum PFOS C_= C. Ct0 C= C= C7 Sample 1, Corrected Sample 1, on-Column for Concentration Concentration Factor 24.859 < LOQ < LOQ 0.313 0.675 11.734 0.250 2.486 < LOQ < LOQ 0.031 0.068 1.173 0.025 Sample 1, onColumn Concentration ULOQ (30.2) 0,177 " 0.760 0.575 1,234 13,396 0.126 Sample 1, Corrected for Concentration Factor Golden West Serum Sample 1, onColumn Concentration ULOQ (30.2) 0,018 0,076 0.058 0.123 1.340 0.013 > ULOQ (30.2) 0.402 2,473 2.025 3.533 26.245 < LOQ Sample 1, Corrected for Concentration Factor ULOQ (30.2) 0.040 0.247 0,203 0.353 2.625 < LOQ Bioresource Serum Sample 1, on-Column Concentration ULOQ (30.2) 1.444 2.856 3.272 2.657 26.245 < LOQ Sample 1, Corrected for Concentration Factor ULOQ (30.2) 0.144 0.286 0.327 0,266 2.625 < LOCi Human Plasma Data, 5x Scale-Up with 20uL InjectionVolume Quantitated with Chinese Plasma CalibrationCurve All data PFOS _'_z C11 C_0 CD C= Cl is in units of n_/rnL Lampira Plasma Sample 1, on-Column Concentration ULOQ (30.2) 0.364 0.486 1.270 3.799 36,207 < LOQ Sample 1, Corrected for Concentration Factor > ULOQ (30.2) 0.036 0.049 0.127 0,380 3.621 < LOQ Golden West Plasma Sample 1, Scruple 1, on- Corrected for Column Concentration Concentration Facto=r ULOQ (30.2) ULOQ (30.2) 0.236 0.024 0.711 0.071 2.112 0,211 3.436 0.344 ULOQ (31,0) ULOQ (31.0) < LOQ < LOQ Innovative Research Plasma Sample 1, Sample 1, on- Corrected for Column Concentration Concentration Factor ULOQ (30.2) 0.219 ULOQ (30.2) 0.022 1.400 2.169 0.140 0.217 4.016 0.402 37.170 0.160 3.717 0.016 Chineese Plasma Calibration Curve for Scale-Updata, 10uL InjectionVolume Analyte PFOS Calibration Range, ng/mL 1.01- 30.2 R= 0,999539 System Suitability Samples, %RSD 4.21 CCV #1 recovery, % 155% Matrix Blanks < LOQ t'_ Ct= 0.104- 3t,1 0.999696 3.94 114% < LOQ tJI CH 0.103- 30.9 0.999526 6.34 95% < LOQ CI0 C0 C= iC_ 0.101- 30.3 0.254 - 30.4 0.103 - 31.0 0.102- 30.7 0.999907 0.999812 0.999441 0.999903 4.02 4,16 5.99 6.21 94% 92% 92% 106% < LOQ < LOQ < LOQ < LOQ ,i :_:i!i _ ! , _'!_i_ili '_" i:::i ' -_i _:iiii !!i_ Matrix Spike Recovery for Human Serum Data Quantitated with Chinese Plasma Calibration Curve Analyte PFOS C12 C11 Clo CC9s 07 Lampire Serum Average Endogenous Concentration, ng/mL < LOQ Low Spike Recovery, % < LOQ High Spike Recovery, % < LOQ ISigma Serum I Average [ Endogenous I Concentration, I ng/mL 4.56 < LOQ < LOQ < LOQ 129% 164% 136% 114% 140% 127% < LOQ < LOQ < LOQ <0.L6O5Q 115488%% 111283%% 01..2670 < LOQ 150% 130% 0.15 Low Spike Recovery, % 39% 90% 138% 120% High Spike Recovery, % 90% 75% 109% 103% 15135%% 100% 111031%% 113% _,nalyte PFOS i Clz IC11 IC10 IIC9 C8 C7 GoldenWest Serum Average Endogenous Concentration, ng/mL 27.02 < LOQ 0.32 Low Spike Recovery, % -573% 92% 106% < LOQ 120% 0.90 5.60 O.19 80% -191% 44% High Spike Recovery, % -19% 73% 1221o 104% 114% 71% 94% BioresourceSerum Average Endogenous Concentration, nglmL 17.02 < LOQ 0.30 Low Spike Recovery, % 0% 530% 93% < LOQ 864% 0.61 2.95 < LOQ 86% 91% 294% High Spike Recovery, % 3288% < LOQ 6% 146% 49% 624% 36% _i I i Page 60 of 225 '_'__ '"_!: _-_t i i :i ,( :,;J :_i ;i,_!!:l Matrix Spike Recovery for Human Serum Data Quantitated with Chinese Plasma Calibration Curve Analyte PFOS Ct2 Cll Clo C9 Cs C7 Chinese Plasma Average Endogenous Concentration, nglmL < LOQ Low Spike Recovery, % 260% < LOQ < LOQ 104% 130% < LOQ < LOQ < LOQ < LOQ 80% < LOQ < LOQ 56% High Spike Recovery, % 96% 100% 104% 98% 98% 94% 106% ILampirePlasma Average Endogenous Concentration, ng/mL 11.12 < LOQ < LOQ < LOQ 0.44 2.61 0.20 Low Spike Recovery, % 407% 96% 92% 98% 69% 147% 64% High Spike Recovery, % 102% 91% 88% 80% 89% 94% 105% Analyte PFOS C12 Cll C1o Cg C8 C8 Golden West Plasma Average Endogenous Concentration, nglmL 18.26 < LOQ 0.10 0.16 0.54 3.87 < LOQ Low Spike Recovery, % -940% 88% 84% 82% 37% -106% < LOQ High Spike Recovery, % 42% 96% 97% 103% 92% 81% 125% Innovative Research Plasma Average Endogenous Concentration, ng/mL 15,58 < LOQ 0.14 0.17 0.59 3.07 < LOQ Low Spike Recovery, % 348% 116% 141% 122% 51% 159% " 1438% High Spike Recovery, % 121% 113% 121% 118% 108% 122% 183% Page 61 of 225 .. : :I '_,:! _;,i_,:i :!=.J i I " ii i : _...:.,_ ::,_ i ,' ! ,..! 3M ENVIRONMENTALLABORATORY E02-1039 PFOS Telomer Analysis Summary 3M Pharmaceuticals Analytical Research and Development LC/MS analysis of a pooled human sera sample obtained from Golden West Biologicals (#E02-1039) was performed using an Agilent 1I00 quaternary I-IPLC system coupled to a Micromass Q-Tof2 quadrupole time-of-flight mass spectrometer. The sample was run as submitted using the following parameters listed in Table 1: Table 1 Inject: Analog wavelength output: PDA Wavelength Range: Recorded Mass Range: Flow Rate: Column Temp: Column: 20 gL NA 190-600 nm 100-800 Da. 0.3 ml/min ambient Betasil C 18, Run Time (rain.) 1:I20w/2raM N_CI-I3CO2 MeOH 0 90 10 7 0 100 12.5 0 100 13.0 90 10 17.0 90 10 IonizatioMnode: Resolution: 50x2mm ESI(-) 7,000 Accurate mass measurements and elemental compositions obtained in this analysis are summarized in Table 2: TabRleete2ntion Time 5.8 min 6.3 min 6.5 rain 6.69 min 6.71 min 6.72 min 6.8 min 7.1 min Accurate Mass Theoretical Mass 362.9739 412.9637 526.9693 462.9613 498.9280 512.9615 426.9734 562.9542 362.9691 412.9659 526.9610 463.9627 498.9297 512.9595 426.9674 562.9563 Mass Deviation (ppm) 13.3 -5.4 15.8 -3.0 -3.3 3.9 14.1 -3.8 Probable FormuIa C702F13 CsO2FIs CI0H403FITS C902FI7 C803F178 C i002Fl 9 CsH403FI3S C1102F21 Page 62 of 225 !.i _,_.,_ ATTACHMENT C: CHROMATOGRAMS 3M ENVIRONMENTALLABORATORY E02-1039 ]:!:ii_i ! ,iiJi: =i" Page 63 of 225 ( plasma up Ig3dMo02l1d0e30n0w05est 100 5xscale 1012310o2k 8.04 8.53 2: 3M0R-OMcto-2f 04D0C2ahvane1ny2e:0l3s77:E04S_7-9S TIC 4.36e4 ' % 1.06 %t O" , , d021030005 .............. 10.26 / 7.45 9.56 9.,96_ 10.7510.96 2" , .... ''' "' " ...... ,1.,, VV-" V! ., 12.51 I ' ' ' I , , i .... 1 .... 1 ' ' I .... I "*"' *" 1: Daughters of 363ES- 15.15 TIC 100 t t 15.23 7.73e7 co 1171 14. _" 12:52 14.80/t I _ O_._ (::)0 0 .... _. . .,'.. . . , .... _ .... , .... , .... , .... _ .... , .... , .... ,...._ .... _ .... , .... ,.... Time _) 2.00 4.00 6.00 8.00 10.00 12.00 14.00 _ ExacCt opoyfOriginal (._:v _ _/_,/__ InilJaJ Date goldenwestplasma5x scaleup 10/23/02ok 3M Environmental Lab d021030005 273 (7.408) Sm (Mn, 2x1.00); Cm (272:274) 100- 74.459 84.685 davey 070799 30-0ct-2002 12:37:49 1: Daughters of 363ES- 2.99e4 362.960 113.088 ] I 169.102 ' J 318.241 c_ '_/: / t il i J 87931" Ill 9.007 _ '-0 1236.154 I i _ L 3 19.373645j420 ;o 391.469 0 _-_ 0 m/z "c__ t_ 60 80 100 " 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 co :o t,J _o-< Exact Copy of Original Initial Date " I! golden west 3M d021030005 plasma Gx scale up 10123/02 ok "_+ 1001 0 ; _ '-'_"1 _; ' "_ ' | '_ " ' I_ ' ' ' ' I '_T_" ' ;-I .... d021030005 I .... Davey070799 30-Oct-2002 12:37:49 2: MRM of 4 Channels ES- 8.01 363 > 319 10.26 10.75 +:_ +:I/__,o_ ' 732 ;36 9.29 i .... I '_''' ' ' I .... I .... I .... 12.05 I .... 6.45 4.09e4 13. 89 12.97 13 16 I ' "_ ' ' I .... | .... 2: MRM of 4 Channels ES- 363 > 219 %- _00-. 7700.81 1,16 230 _ d021030005 7.82 - 10.29 _-=_986p5 _8;I/_'!-'_11;_r'I.-4151i6192:4+_1272 7.40_ 15.21 15.78 2: MRM of 4 Channels ES- 8.5: 363 > 169 1001 7.45 " 3.76e4 +o, ._..1_ -, __ ,_,,.+_.+_,,,,..,-)_gA,:,,,+,,__+_,_,_..++_ +;, ..... t .06 0.84._, _11.62+ __2.17 2 68 ............. " 55 7.17 ......... 9.56 9. i .... ,10...3. 4 1I1..2..1. 11.8=8.. 12_0 ; ,. 1!_4..27 , 1,,5_..2_1., co .+ +,'" n_:_, d021030005 2: MRM o1'4 Channels ES- 7.45 363 > 119 _Pq 0k._ t_ t._ t.,,rl % 7.17 II 1034 |.__./_ _84 1.19 0 .... _, , _, _ 2.00 - ._p1 ....'...I. ' ExacCtopoyf0dginal _4.7_7 523 5.90 66- 0 I _ .... + .... _ .... 4.00 6.00 8.12 _ .... 8.00 9.10 9.83 -1067 11"611.75 _ .... _ .... _ _ , , , _ .... 10.00 12.00 13.02 133. 2 _ .... _ .... 14.00 149._1...._15.35 -. _ .... llme r- __ O _ _ :o l.o ..< golden west 3M d021030005 100- plasma /" 5x scale up 10123102 ok @% I%- 114;) ' 8.0' 10.26 10.75 9.96_ I 10.96 o-_--_.,....-, ._.. ,-..--.,_. ;-,.... , .... , .... , ._.._,.Vt,.... d021030005 100 8.45 ,.___._,. Davey070799 30-Oct-2002 12:37:4_ 2: MRM of 4 Channels ES- 363 > 319 4.09e4 _..-,.._-... 2: MRM o14 Channels ES363 > 219 oj d021030005 100- 21 ,o, 8.53 7.40e3 ......... 2: MRM of 4 Channels ES- 363 169 3.76e4 7.45 %. 1.06 o ..0.....8..4__/ ,"___;.1._.-.1_"62,"_.2.".1.7268,.. ,_..,. ,-_, JI755 9.56 717 ..._.___, .... , .... , ....'I'._.9_.3''7 /I 10.34 '"l_l__.."'_', 11 21 11.88 12.80 ; 14.27 15.21 .._ .,-._. (,a _, '";1' " d021030005 2: MRM of 4 Channels ES- :_ 100 7.45 363 > 119 o _ o.76 _ .72 8 12 9 10 10"_10;67 li69 _-,-,I _-'_'_]|_( _._.1.19 3.01__ .,..i..77_.235.90660flll, l 2 . 9.631 I " " 13.32 I _,1175 1302 14._1 / .... ,,.._,,-.15"35 O 0 .... , ........ , , .... , .... i .... I .... , .... , .... , ...., .... , .... , .... , .... , .... , .... Time to 2.00 4.00 6.00 8.00 10.00 12.00 14.00 i',O - ExacCtopoyfOriginal _0 _(_ _ :o _D-.< golden west plasma 5x scale up 10123/02 ok 3M d02103000_ 1 O0 goldmn west plasma Sx scale up 10/23/02 ok 314 Environmental Lab 7,,, _ , =as dave0y70r99 30-0t-2002 12:37:41 z,,4 ==m . G0ZI0_D05 213 (7.4011)5m (Mn. 2x1.D0);C41_l T2:ZT4) 1; D_Jiht4_i Ill 3_1E8- o/0_ 8.01 10.26 10.75 9.72 _,"/_ /_11.07 Davey070799 30-Oct-2002 12:37:49 2: MRM of 4 ChannelsES363 > 319 4.0904 0 d0210300( 100-1 "! .... , .3._ ,. 1= II i I I m.,_ I ===._= =1=.=,1 I =_s.,= I 8.45 I I 2: MRM of 4 Channels ES363 > 219 7.4003 o d ........"_'""i" ........_.-......._ ......._._...._....._ ....... _-_- - %]_6' d021030005 230=_ 3.474.04 12 5.31 598 "c'_'85 ::_ 26"t'_ 9.16 10.121029.10.45 il.69 100 8.53 .... ... 12.48 12.72 1_.78 2: MRM of 4 Channels ES- 363 169 7.45 t 3.76e4 J 1 92217 %01 ,0, .84.._'_1.62 2.6,8, ,-.,, d021030005 100 ,,.'_,...;-,_,':-; " 7,l_,.,J,._, ,_,,_ __ 7.45 ., 14.27 15.21 ;_" 8- - .-, .',-,-_.., ,'4'...:. : _i,' 2: MRM of 4 ChannelsES- _, 363 > 119 "_o t 0.75 % 7.17 t i0.84 oo o - 72812 ,11i6 10167 "',_ 5.23 o.=_. 6.60 /_l_l ?'- .9"xi09.83 _ j[ _. , , _ ..-.'.__ z .... _ .... a_ ..'._. ",_-__".,.,.,.t..n"__._..,..,_,_,.,_,,,.._..,.... _ .... :3i02,13.32 14.91 15.35 z ......_ ...... "...... ,,_.A..,._.,./i_rr-z.e bO _ 2.00 4.00 8.00 8.00 10.00 12.00 14.00 _ r-- 0 ,_ co:_ _O-.< ExactCopyof 0dginal ("r_ I_J._ .I _ ::L. Date 02048-21-7 3M solvent d021030015 100- standard 10ppb Cw-a&JS 7.36 Davey070799 30-Oct-2002 15:29:24 2: MRMof 4 ChannelsES- TIC 2.29e5 l%. 0 .' "; = _"T';"''_'"-Y'_'_-'--F d021030015 -''-_-' i 11 I ,,_......._..i__..,:_..D_.,,__._-,___- 7.15 % 1, i ' i ' ' ' i ' ' _ ' '"' ' g r, 2.07e77"33 7.16 1_ ,)_k' ,3 .... i _ 1087 ; i .... _ , ; "r"l 1: Daughtersof 363E8- 13.44 TIC 1228 A ",. 1274 I_ 14.37 15.28 .... ,jw_ _-, vv_ '_._'7_J__,/'._'_ _.. 0.88.1.02 210 570 6.70-- "_,ll 7"95.8.14 9_01 ".,_11.74-_ l.'_ |1,_1...,.l_J*.,_,,..t _, _l1.(..]o._:_ _ _m_O 0 .... , .... , .... , .... , .... ,,-... , .... I .... , .... , .... , .... , .... 0'...., .... , .... , .... ,Time ,_ o 2.00 4.00 6.00 8.00 10.00 " 12.00 14.00 16.00 _ t,J (,o:0 ExactCopyof Original ,. _"_ Dato .---.- ..... ._,: ..... _-- -_.::---..... .... ._:_... 02048-21.7 solvent standard 10ppb 3M Environmental Lab d021030015 270 (7.325)Sm (Mn,2xl.00); Cm (260:277) 100 168.504 I I davey 070799 30-Oct-2002 15:29:24 1: Daughtersof 363ES- 5.29e4 319.257 118.887 _ ;_-; 318.241 \ 61.959 t_ ...j 237.110 280.606 300.977 rT1 _O 0 60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360" 380 400 _ I'J co:_ ExacCt opyofOd_!nal InltJal Oate ....................... t .' ........ _.--_..._.......... ,....... 02048.21.7 3M solvent standard 10ppb d021030015 100 7.3, 1%t ' d021030015 _! 100 0.89 ot | o.32 ; d021030015 100 %-t 7.17 - -_= " - 7.3, 7.15 Davey070799 30-Oct-2002 15:29:24 2: MRM of 4 Channels ES- 363 > 319 .. 1.91e4 2: MRM of 4 Channels ES363 > 21_ " 1ti72 12.83 15;37 , " ' 2: MRM of 4 Channels ES- 363 > 169 1.66e5 0 ".. ' i "'" ' _ .... : .... I .... i '' I .... d021030015 100 %'" I .... 7.34 7 ' ' , _ 1 _ ,', ', "_". , ,'i ,' , , i .' . ,',' i . ; ._ i' . ,'. , i 2: MRM of 4 Channels ES- 363 > 119 7.12 t'n.. _" ;* .:_ o 0 :, . , _ .... . . . _ _" , , , .... .... ; ....... .... r .... .... ', . : .; .-; . ,-.'_. Time ,_; 2.00 4.00 6.00 8.00 t0.00 . 12.00 14.00 ii 16.00 i _ l-,.) t./) ExactCopyof Original _o-< ,. I O2048-21-7 solvent standard 3M d021030015 4 _rt 0204S-2t-7 solvent standard t0ppb 10ppb ; UU-[ 1 3_M21E03n0v0ir1o*n' m2e1n0t(a1l.32L_a)bSm (Mn.2x1.00];Cm(Z60;277) daveoyToTm 7.34 =o-o,_ozl=:z=:z4 I I; Oevihle(sof36_E8- | Davey070799 30-Oct-2002 15:29:24 2: MRM of 4 ChannelsES363 319 " 9_ 4 1. 1 e m% 7. 0t_..2.oM1.c/ d0210300" I 100;' ,,,,I / - =a_-T ,,,.,I ''1 .... , .... _ .... _, .... , .... o .... , .... , .... ; .... , 2: MRM of4 ChannelsES- 363 219 2.80e3 % .L-':i_.o....i._......_. .....i._. ":=:: '- ,. ....,._. "s .......... 032 ltno_ 192 \ _ 349403 ":'v 552 "';" 898 _._o -7.77_9 J.... i u.o] lrJ.:_9 1140! ]-'Jo I la 97 ls.os ',_I2g /. I 0 ' ' ' ' I ' ' ' ' I ' ' ' ' I ' ' ' " I ' ' ' ' I .... I ' ' ' I .... I .... I .... I '"" '" ' '1 " ' " ' I , ' ' I .... I ' ' '! ' ' ' I d0:_1030015 2: MRM of 4 ChannelsES- 7.36 363 169 100-t 1.66e5 ]Id02103..0..0150il . .'. . , .... , .... I'. . . I .... , .... i :-. . . . i. .-.; _ _. ; .1 :.-. .-, .... i .... 2:MR.M"I'of'4'lch"a'nn'e,ls"ES- _ 734 363 119 7.12 0 ". _.. , .... , .... , .... , .... ,..... , . . . . , . .L__._...,.. , .... _'_.-. ,-..-.. , ._.. ,-..- , .-; . . ,-..-".. ,Time ,_ O_'_ ..... 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 -c___) ExacCt opyof Original ,,- ! ,_.1'1'3(_. t/_/o._ ' Dais JR,/ goldenwestplasma5xscaleup10123/0o2k 3M d021030001 100- 8.37 Davey070799 30-Oct-2002 11:28:58 2: MRM of 4 Channels ES- TIC 5.78e6 |%. d021030001 8.08 8.40 1: Daughters of 413ESTIC 5.51 6.09 L I, _,t 8.98 10.43 12.21 _: _: 0 0oQ_ 0'--_ .... ,Time "" _"_" 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 co ::o I_ to-,< L,/z ExactCopyof Original golden west plasma 5x scale up 10/23102 ok 3M Environmental Lab d021030001 288 (8.395) Sm (Mn,2xl.00); Cm (284:297) 100 168.768 .o davey 070799 30-0ct.2002 11:28:58 1: Daughtersof 413ES- 8.08e5 % 218.992 369.289 ",J' ;[. ::o _ 118.651 ] _= _ 413.208 _ Om _o 60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500 "_ tJ -- _;0 ExactCopyof Odginal Ci_t _o _._j__. Dat .. golden west plasma 5x scale up 10/23/02 ok 3M d021030001 100 - 8.3 Davey070799 30-Oct-2002 11:28:58 2: MRM of 4 Channels ES- 413 > 369 = %-I 0 '1 ' .... i d021030001 100. ' ' , .... I ' ' ' i ' .....'"' _ 5.21 e5 , ' ". ......i' " 8.08 % ._. L 8.37 _I , 0 0 _i _ , . I 0 i 0 o I 'P"_' ' ' I "" ' ' I . 0 I . , I 2: MRM of 4 Channels ES- 413 > 219 9.47e5 %- 4 d021030001 100 7.798.05 \_.._/_L_ _ .,,.,. 8.3' , .... , .... , .... , .... , .... , .- 2: MRM of 4 Channels ES- 413 > 169 4.09e6 CO 0/01 8.11\ ./: Plr_- 0-' .... i .... I .... i .... , .... l .... l .... I .... I/''P' _' ' I .... l .... l .... i .... , .... l .... l .... l _ ii: d021030001 100 2: MRM of 4 Channels ES- 8.37 413> 119 . o 2.268S o_ c0 8.17 _" ._j co o - . _. 0 I .... , .... 2.0, 0' "" ' , .... 4.0j 0.... , .... 6.0,_0: ' ' , .... 8.0,'0 _'"1 .... 10.,00.... , .... 12.,00.... , .... 14'.,00.'... , .... 16.,0T0ime _) (,o ::o ho (o,.< Exact Copyof Original :_:__: _ i ::i__ ..... :_._! _:_:_ __::i : _....%..:__ ..... :_ : ...... golden west 3M d021030001 plasma / 5x scale up 10123/02 ok 1001 %-I 3M Envb'_msBtal Lab dD2IO3000t 288 (8 _a5| Sm(Mn. ;b;t.00_;Cnt (214_=1) 100 I_.?U golden,,,,t p",,= =*,,l, upI=Z=SoOk= _.Oct-200| tldlll:6B 1; Dm_ht_l d 413ES1.3g"at d.,,yOToT=" 8.37 " Davey070799 30-0ct-2002 11:28:58 2: MRM of 4 Channels ES- 413 > 369 5.21e5 d02103[ _. 100-, 2t Jim2 ...,,,.,,8,..08,,._.L.., .... ...,...,,.,.-,0.,._..,.... , .... ,.... , 2: MRM of 4 Channels ES- 8.37 413 > 219 9.47e5 ,=.......i........_. ........ _" ..-i_.._.. d021030001 . i .. . , . , i o . .'i ,l , 8.37 2: MRM of 4 Channels ES413 > 169 1001 4.09e6 8.11 ._, 0/1 n_ .... _.. , . . _ .... _ .... _ .... _ .... _ .... _ .... _ , _ .... _ .... _ , , . _ . , .... _ .... _ .... _ .... _ i.: d021030001 100 1 2: MRM of 4 Channels ES- _ 8.37 413 > 119 0 " 2.26e5 _: IT; 0_ 8.17 _" ",-J o c_ '-_0 0 _ .... , _ .... '_.... _ ,', , , _ .... _ .... _ ... . _ .... _ ' ' ' i ' '-. _. , ' . _ .... , .... _ .... , .'_'.'. _ ,'Time _ 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 _ :_ Exact Copyof Original j.ol_,/c:z_. 02048-21.7 solvent standard 10ppb 3d0M21030011 100- I1%, .o Davey070799 8.3; 2: 3M0R-MOcotf-42C00ha2n1n4el:s21ET:SI0C-4 6.47e5 0 ",", , T I .... d021030011 1001 I .... I .... I .... I .... I .... I _..... d ' I ' ' ' ' I '=; ' '"_"_;- 8.34 ' "'1 _" ", , ]-, , ; ! T , , , ! '-" ";--;" I 1: Daughters of 413ES- TIC t i ._ 1.24 t Hs: 46 10.11 11.45 11;9512.09 .... 1404 =o./_ 14.71 3.81e7 _ r_ g -.a _o "oJ 0 .... , .... , .... , .... , .... , ..... .... , .... , ..... ',",,, ,". ,,., .... ,...... , .... , .... , .... ,Time ,_ _._ 2.00 4.00 6.00 8.00 10.00 12_00 14.00 16.00 _ _) t..o _o.-< ExacCt opyofOriginal ,,. ! _ 020--_-2-1-7- solvent standard 10ppb 3M Environmental Lab d021030011 286 (8.339) Sm (Mn, 2xl.00); Cm (281:288) 100- 168.768 davey 070799 30-Oct-201)2 14:21:04 1: Daughtersof 413ES- 1.90e5 218.255 368.797 co )-18.931 0 119.082 4 _0 58.794 0_, ................6..0....... 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500 "_ "_ _0 tx_ Co::O O,< ExacCt opyof 0ri0inal ;-" Initial Date ........... _ ...................... _-". ............. ---'."... __: j _ !>-_i _ _7_ _ : i__-i_ __-_'_ 02048-2t-7 3M solvent standard 10ppb d021030011 8.32 Davey07079 30-Oct-2002 14:21:04 2: MRMof 4 ChannelsES413 > 369 I% 10001"_-,"_-;'"'l_- J;-";' I_ _ d021030011 :': 100 6.13 a" -' "-'_ ...._, , , ^, 8.67 9,28 _''_'_-i_'_'__ 8.32 %- ; _-_'..-:5'.9,6e4 2: MRM of4 ChannelsES- 413 > 219 9.42e4 0 " '' ' I ''' '_' I .... d021030011 14 1 1 ' ' ' I , ,'I , i,, ,'_ I, , ; , i , ,-, t'_-_ 'L_-' * t=' ' _'l t .... 8.32 :I--'-' ;''-I '-i ;-' I ' ' ;_-|';'''_'%_''' I 2: MRM of 4 ChannelsES- 4134>.631e659 d0210.3.0.01.1. , .... ,.... , .... ,.... ,.... , .... , .... _o...., .... , .... ._......,.....,...2.: M,R..M.o.f 4,C.h.a.n.ne,lsES- 6.32 413 > 119 1 co _,_,-mi' -,4 8._ 03 12.19 = _-_ 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16_.0-_.0... _ L_ ExactCopyof Orioinal / gk_ f_ 02048.21.7 solvent standard 10ppb 3M d021030011 100 o2048-21s-7olvensttandar1d0ppb 3M Envkonmental Lab 002103001 "_ 286 15.33g) Sm |Mn, _:x1.00); Cm [281:2t_) 100 160.768 % davey0707SS 30-Oct-2002 14:21:04 '$;Daughte_ _| &I3_ES- 1._3e5 8.32 d02103001 _ _,.. 1001 218.255 :3M,TP7 8.67 9.2S 8.32 Davey070799 30-Oct-2002 14:21:04 2: MRM of 4 Channels ES41 3 > 369 5.96e4 2: MRM of 4 Channels ES413 > 219 9.42e4 %t .s=_ 5.s.7=l4 f sO..._.'I_ 0 " ' ;'1 "'" ''' I ' ' d021030011 100 _218.931 t ...... , '_0 ' 2_o' _" 3_0'4_o '._ .._.o."._ 'I; ' ' I ' "'''_ ' I"' '"" I" ' + ' I " ";-" _ 8.32 - ^ 2: MRM of 4 Channels E$- 413 > 169 '%]0/ ,., .... d0210.3..0. 011 4.63e5 ] -. , ! ,1 ,,, :,, - .,. ,,, ,,,, i __ ,, _. ,, i.,,, 6.32 l,,, ,-i, ;,.,, ,'. ,'i'., ,2:M, RMi,..oi Cf4h,' a'n'n'elsE'$- 413 > 119 _m=; l _o 1001 8.0\3 } _ OO 0/ , , . , .,......... _ .... _ , , ,-. _ , r'. ,_ , .... , , , _-.. "_" . ,_,"_.. 1219 ; ; , __ 3.02e4 .":-,-'_ Time _ oo ,_ ,-_ ...... 2.00 4.00 6,00 8.00 10.00 12.00 14.00 16.00 _ ExacCt opyofOriginal ,' ae up I3gdMo02l1d0e30n0w06estp"lasma5x scale 100- 10/23102ok I1%- 10.06 9.21 2: 3M0R-OMcto-2f 04D0C2ahvane1ny2e:0l5s75:0F0TS27IC-99 8.32e5 d021030006 100 10.02 1: Daughters of 463ESTIC 11.04 A _ lO.89 . 12.o9 _,'" _: A12.20 14.35 14:87 7.86e7 % 9.21 ; _ 11 47 8.68 J_ 9.70 / _ -ii 8.76 _ o ,_ _, ....... , .... , ...... ' .", .... , .... , ..... "..-.'t'...., .... , .... ,'...., .... , .... ,Time _ 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 co :_ t..o _o.< ExacCtopoyfOriginal InlUaJ Date 0 golden west 3M d021030006 100 plasma 5x scale up 10123/02 ok 10.06 Davey070799 30-Oct-2002 12:55:02 2: MRM of 4 ChannelsES- 463 > 419 d021030006 100 9.21 " A 92.1 8.28e5 2: MRM of4 ChannelsES463 219 2.83e5 %1 t o].... ,.... ,.... ,.... ,.... ,....,.... ,.... ,..._'__,.,.-,..,... ,.....,......,... ,...... d021030006 2: MRM of4 ChannelsES- 100 9.2 463 169 3.06e5 %1 9.01 .,. d'_-Jh''1''l'I'r l I i i i I 1 4 6 1 | i i i i l l l l i 'I i . , i I ' i I i I i i , i I ' ' n "_ h , ''I" ' ' ' i i ' ' ' ' I" ' ' ' I'' ' ' I ' ' ' ' I i , , '," d021030006 2: MRM of 4 ChannelsES- 9.2 463 > 119 CO _._;I 3:o o 1001 1.36e4 r_ /-J 0.84 r- __ _.oo ,.oo _.oo _.oo lO.Oo ,_.oo ,.oo _ cD 01 8.98 oo 6.80 15.32 t_ 0O_ 0,1,-;--:-T_-.,= ......... , .... 3.4,q.3.57 , ....._ _ .... _, "= _ = ,, = i , 10J.03,'1-,0-_7-6, 11.48 =12.33.1.247_..,_ 15.20..._^ Time IcT_I_0" o,.< ExacCt opoyfOriginal In_al Date _:_i _ i i_ _ golden west plasma 5x scale up 10123102 ok 3M Environmental Lab d021030006 10C 316 (9.206) Sm (Mn, 2xl.00); Cm (314:321) 168.522 218.992 _ _*_ _' _ davey 070799 30-Oct-2D02 12:55:02 1: Daughters of 463ES- 8.07e4 268.835 418.552 462.836 160.222 \ 119.105 / r._ 444.964 0 m:_ 279.220 329:050 362.654 401i169 tI _o I,O 60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500 __,,_ ExactCopyof Original cgo,-p '_0 t-,,J %,re ,<' golden west plasma 5x scale up 10/23/02 ok 3M d021030006 1 00" golden west plasme 6x scale up 10/23/02 ok 3M Environmental Lab _021030006 316 (9.208) Sm (u_. 211 .O0);Cm (314:321) 100- 1M 522 2111.(i02 clayey0707. _ct.2002 t2:55:02 I: Diugl_em of 43ES- 8"02ro4 I1% 10.06 DaveyOlO799 30-Oct-2002 12:55:02 2: MRM of 4 Channels ES- 463 > 419 8.28e5 _ 9_A21 0 _ ._ llm -, . ; i- ._. i .'-r--_., 100 _i.lm 9.21 463 219 %- I 1E0.222 1111ES1 33050 401.Ill ,Ihi, ,11 , _41i..'!_!. I i, I J 2.83e5 I_o l_o 2oo 2'_o 3o0 _o 4_o 45o 5oo 8.9_ 0 J , , , "=i' I d021030006 ' ' " ' I "' " I' '' ' I ' ' '; ' I ' ' '' i ' ' ' ' I ' ' ' " i " " ' "_ mi' ' "' I'" 92.1 '" ' ' I ' ' ' " ( ' ' ' ' | ' ' ' I " ' ' I ..... " 2: MRM of 4 Channels ES- 463 169 1001 %1 0|' ' ' f" I_' '''' ''I '' '' I''' d021030006 ' I .... I * '''I " * _'' I''"''' '1 01 0 -=.... :. .-', , i- .. ; .... " ...... 2.00 4,00 680 ., , .' 6.00 9.01 i'' ''' _""' ''l =' '' ZI''' 9.21 t898 II 8.00 10.00 ExacCt opyof Original Dat_ 3.06e5 .._, ' I"" ''' i '' ' ' ' I .... I .... 2: MRM of 4 Channels ES463> 119 . 1.36e4 12.00 14.00 15.32 Time ___: c_ _o o ;_ r_ o _ _ :o r,o ,..< C--c/ 02048-21-7 solvent standard 10ppb 3M d021030016 9.10 100- Davey070799 30-Oct-2002 15:46:28 2: MRM of 4 Channels ESTIC 4.98e5 I1% t" O- ,"; ";-_-i _'. ; i ' "" i .... . d021030016 I "-' ' I ' ' _ ' _ '"" ' _ "'" ' J ' ' """' _ ; "_: 9.09 i ; ' ; T_--;--;-.. j ,,-'r"_-;--T_'%',--r-_"-:-]-'_--,-_ 1: Daughters of 463ESTIC 1 100 ] ' _/ 9l .24 1.1_6 1_2_ .... ..60.... 3.32e7 6.62 J_/ 11.08. \ 12.2'! "-;" 14;_l:S/ |_;o| 1' , oo ' _0 0 .... , .... , .... , .... , .... , .:.. , .... , .... , .... , .... , .... , .... , .... , ...., .... , .... ,Time ,_ o __ ...... 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 _ I.O _o..< to Exact Copyof Original Orn( I I/_/O'Z._ --Ini_i___ " Date .... -, ..................... ," ".... _ ....... ........... - 02048-21-7 solvent standard lOppb 3M Environmental Lab d021030016 1 ]0- 312 (9,095) Sm (Mn, 2x1.00); Cm (308:318) 218.869 218.25.=. % davey 070799 30-0ct-2002 15:46:28 1: Daughters of 463ES7.22e4 168.399 o/ 268.896 418.982 _:/ 102.598 152.659 443.305 :o 0 _-_0 t,J 60 80 t00 120 140 160 180 200 220..... 240 260 280 300 320 340 360 380 400 420 440 460 480 500 _ CO-< ExactCopyof Original dauL " k_al Date '_ ........ _'*'-.................. p ,'" :"'-" '-"- .......... _" 02048-21-7 solvent 3M d021030016 100 standard 10ppb Davey070799 30-Oct.2002 15:46:28 2: MRM of 4 ChannelsES- 9.10 463 > 419 [] %-t i 1.13 Ol:T """' I"" ";-;":;:1 _" d021030016 ' ; i ":' ,.2, ...... 1*_' -| `% ' ;'i ' '" " _t_'- ,o.oo,.,, ' " "- I : ' '- . i] .... I .... I .... 9.10 _ 1001 % 6.95e4 12.31o3.21 1 .... l .... _1 ' ' ' '_/' '--%''; I 2: MRM of 4 Channels ES463 > 219 2.04e5 C I '_:j ' ''--;'lJ .... d0_1030016 100 1 I .... ''l , , , i i _ , , i j .... ill .... jlll'] ' _llijIl ill ,:, J , ,q, j .... ] .... Ili,l, , iII _ .... J 2: MRM of 4 Channels ES- 9.10 463 > 169 2.18e5 %1 d02103.00. 1j60'"_' ' ' I .... 100 I " " ' ''I ' "' I ' ' _ ' I ' ' ' ' I , ". a , I " ' ' ' _--' 9.07 ' ' I ' ' ' ' I , . , _n. . , , I ' ' '2_' MRM' ' ' 'of'4' Ch" a' n'n'elEs'S463 > 119 c_ _ i_ :_ _ /ot 9.2_ 105_ 1402 __." D_ o o{._,_..--._-1:..6TB,-_..... ," ........., ....... s,_ 6.14 ". , .... , .... 7.24 s.23 , .... , ........ ,-......, ..._....o....o._.,...._....... , .._..2.s,7..'1.'._,.0........_. . .-.,'..._..1._5.._.-L-,T4ime m ,Ip o 2.00 4,00 6.00 8.00 10.90 12.00 14.00 16.00 _ F,J ExactCopyof Original Ir_tial Date -_:+; _@_ i _-, ' _::_+_ ()2048-21-7 solvent standard 10ppb 3M d021030016 1 00" zo4s-2_-7_t _=td lOpp_ 3M Enviroclllrlenl, al I=b _(32103301e 3t2 _ 0;_") Sm (M_. _t .CO); Cm _30&318) 100 21Szu_ claye0y7_'tm 30-Od-200t2S:4=:=e 1: _u_h_wz d 46_F_% 73_e4 ! d021030016 _ 100 '_ _._ 0/to o] d021030016 100-_ . ,.. _,.o.J_......a L___L- i 4_,.,..,._ 50 I00 150 200 250 300 350 400 450 500 . 9.10 _ 140.15 9.10 Davey070799! 30-Oct.2002 15:46:28 2: MI_M of 4 Channels ES463 > 419 6.95e4 12.30 13.21 14.14 2: MRM of 4 Channels ES- 2.04e5 2: MRM of 4 Channels ES463 > 169 2.18e5 0 "I_+ ' + l ' + ' ' i ; + ; +"i t', d021030016 100 , , I ' ' ' ' l +' _ ' i + ' ' + I '-_ r:, i ' _ =_ ' ' [ ' i ' ' ' ' l ' ' ' ' i-- _ 9.07 _ [ ' ' + + i .... i ++ ' ' ' i 2: MRM of 4 Channels ES+ 463 > 119 7.74e3 co P'PI+. ;I (_ Oo 0 F.) .... 0 -x .... _ , , , _..-_ .'_ ,', , , _ 2.00 4.00 ExacCt 0_y nf 0r!_;q_! initial [m+_+ _',, , _ .... 6.00 7.24 , , , ,'",'_ .... 8.00 19_25 10:53 _ , .+ , _ .... 10.00 11.5812.57 _ ,'. , _ .... 12.00 13:50 _ .... 14 02 ,,_-,,.v _ .... 14.00 15.54 _ .... ,lime 16.00 ". .._......... _ ........ -_--.-:..... : .,- I .... .-+ _" _0 _ "_ golden west 3M d021030003 100 plasma 5x scale up 10123102 ok I1%- Davey07079fl 30-Oct-2002 12:03:28 2: MRM of 4 Channels ES- 9,88 TIC 3.59e5 d021030003 1( O 1: Daughters of 513E$- 13.46 13 64 "" TIC 1.94e8 7.46 11.92 ('D DO ..iv,.- ._ . -_ _ _ o ..... ,..--.. , .... , .... , .... ,..'., -..'. , .... , ..... , .... , .... , ...-,'... ,...'.r.'...', ... ,Time 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 ExactCopyof Original Init_l Date CO o (3 '_ _ _ golden west plasma 5x scale up 10123/02 ok 3M Environmental Lab d021030003 327 (9.869) Sm (Mn, 2xl .0C); Cm (325:330) 100- 169.19 218.739 268273 268.870 / 194.714 davey 070799 30-Oct-2002 12:03:28 1: Daughters of 513ES- 6.65e4 466.288 (69.303 / 513,307 512.651. 413.53t ,_ 319.229 ;;'_ 422250 \ "_ 279.504 349.213 112.331152.45; __ _ 495.097 r.,- ' " " ' I .... I " _ " ' I ' " = ' ] '-' _ " I ' _ ' _ I ' ' ' ;It . ' z'' I v ''v v I1 ' ' = I " ' I'1'"" w I ' I'1 r'm'g I J * ' _ I _ ' ' ' I'l" ' _ I ' = = _1 ' " ' I " ' ' _ [*'i" " lm/z '-_ 50 75 100 125 150 175 200 225 250 275 300 325 350 375 400 425 450 475 500 525 550 _ Exact Copyof 0dgina! Iniliai Date golden west 3M d021030003 100 plasma 5x scale up 10123102 ok DaveyO7079S 30-Oct-2002 12:03:21] 2: MRM of 4 ChannelsES- 9.88 513 > 469 9.10e4 0-L_'_;'i-I "' ' '"-,-' ' ' ' I `_=' " ' = '=' '_ , ' ;-;' I ' ' ' I ;-; ;_'3 -''`;=; f':_" "_ i .... i .... = .... _ .---'"=. ,~,-'-._. = ,_';.=.. d021030003 2: MRM of 4 ChannelsES- 100- 9.88 513 > 219 1.64e5 %- 9.68 0 ' "' "1 '"" ' ' i .... (1021030003 I ' ' _ ' I " ' ''' I '--' '' I .... I .... I " r '% i ' ; 'r'- ; ' ' ' / '-' 0 ! .... i , " , i ,,-, ,--i .-,, 2: MRM of 4 ChannelsE_- 100 9.88 513 > 169 9.64e4 0/01 0" ......... d021030003 100 co "./, ", ' I" ' ' " I , ' ,'" I'' ' ' i i-.-, I ' '-'-I -'-'_-' _ I =,-_'-_ _T. '-.-i " :"-_i .... I ''; . ' i .-.'. "_l^- . h 2: MRM of 4 ChannelsES- ._ 9.66 513 > 119 o 6.92e3 ._ 1 o_ _._ 4 /9.76 4.66. _,5.15 -,=_ 9.33...._]_ 10.48 11 39 1275 15.4615;67 o o 0 .... 7.-. ......,... ..,. .," .'x_. , , ... , .... _ ...-. ..... ,',.. _ .... , .... , . .. , .... , . . .. , . ;.- .... Time _k3" 2.00 4.00 6.00 8.00 10.00 12.00 14.00 _ _;o !_ o-K ExacCt opyof Original lnit_ Date golden west plasma 5x scale up 10123102ok 3M d021030003 100 golden west plasma 5x scale up 10/23/02 ok davey 070799 9.88 3M Environmental Lab 30-Oct.2002 12:03:28 Davey070799 30-Oct-2002 12:03:28 2: MRM of4 ChannelsES_13 > 489 I 0021030003 327 {9.869}Sm [MrL2xl.00); C_ _:_30) 1:4__u.2__htm o_513E_- 9.1 084 2187392'68.273 %"] 0'..._; I-'-' d021030003 % _ 7_ .z,r_87o- s.:_m 1_0.-0"-8 "_'-z-c'-'-_' __" 513.3o7 9.88 11113.3001_11.8481_-2.'_47 ; ;',- I-, . "_"I ;_;-:'-" 2: MRM of 4 ChannelsES513 > 219 1 413.531 =tj=2= | 1319.5_ _2.717 I_ 2 ] I...... h ..... L I. I, o_L._,...:.,.L,:...I..,..;...,:.I....,.,......-....._...,..,.7......-............_.... 0 .... i , ' _ _oo ?_o z_o _o =o. _o .4oo _ _oo _ r_ d021030003 9 , _ . , , _, , , , , , , , = .... 9.88 1.64e5 ,.,_ , , _ .... _ ,-, , , _. 2: MRM of 4 ChannelsES513 169 1001 %1 0.87 1.05 o'...._,_.-.._.m.,...,-..,,,.-.,-., cl021030003 100 9.64e4 8.43 15.09 ";.Zc-- , ,,,_,,-,-,-,_,_,,_.-._L_-_,:.-,-:,....,_._.,....,,.....,,,,,.. _, 2: MRM of 4 ChannelsES- ;o 9.8B 513 119 (3 9.76t " 6.92e3: t_ \,_.=, .,_u7.59,_. 12.75_ 1.5..,4. 6 15.67 _ ,t-__30 0 ,_,',',.r,., _2.00 ,_,_',,_,',4,..00 .... 6,00.._.. ' .... 8.00''''_'""'"'"'';'''10.00 12.00i'-'"'_ ...... 14.00 "'' _ .... Ttmel _._::0 Fo _o-< ExactCopyofOriginal Initial Date 02048-21-7 solvent standard 10ppb 3M d021030013 100- Davey070799 30-Oct-2002 14:55:16 2: MRM of 4 Channels ES- 9.74 TIC 4,23e5 |%_L O- -;-";-_-_. a"" "-i ' "_ ; ' " ,-I ' ;-a .... d021030013 100- "1" , , ; I _' ,'-' i _ _ _ _ , ''_' _-"- 9.77 ' ' i , _-_:---1_-, _ I ; ' ; 'l_' " -'l , "_"m 1: Daughters of 513ESTIC 4,17e7 ._,,..._...... ___ __ 13.28 8.92 10.16 10,91 12;08 _Jk-..,_ _ 13.83 1467 ..... ,;.ZL,-,_--..__ OQ _._ 0--_ . . , .... , .... _ .... _ .... , .... , .... _ . .'. ,'. ..... , . , . , '.............j ..--, , _- . ,', _Time _ 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 ExacCt opyofOri9in_ D_ "-.-. ........ _ .............. _c ..... ._-',._._ .... ,'" *'".- :_.... 0204B-21-7 solvent standard lOppb 3M Environmental Lab d021030013 324 (9.765) Sm (Mn, 2x1.00); Cm (322:326) 1(30 218.799 268,990 davey 070799 30-0ct-2002 14:55:16 1: Daughters of 513ES1.10e5 o_. 1_,354 168,892 / 118.61(} 319.169 ;1 N r"- 0 60 80 100 120 140 160 180 200 220 240 260 280" 300 320 340 360 380 400 _ FO co'< ExactCopyof Origina! rt" Initial Oat_. '_ ............. _-.' .................... _- ......... ........ -_ _ _ _J_ _ _ .... _ _____ _ i _ _ __i _; i__ r 02048-21-7 3M d021030013 100 solvent standard 10ppb Davey070799 30-Oct-2002 14:55:16 2: MRM of 4 Channels ES- 9.74 513 > 469 1.24e5 "%1 0 I .... I .... i .... _ ' ; '-i-' ' c._i ; ' '-" I-' '-' ' I _' '-'- I; ' "/_-='_'_ _l-; ' ' ' I ' ;-' '-I " ; ' I _ ' ' ; i , '-' ""i "' ' i d021030013 2: MRM of 4 ChannelsES- .'; 100 9.74 513 > 219 0/01 0 d021030013 100 1.68e5 2: MRM of 4 ChannelsES- 9.74 513 > 169 1.22e5 %1 d021030013 2: MRM of 4 Channels ES- 9.74 513 > 119 r.o '_ 92 r_ 0Q 9.59 _" 8.333,.,3,_ 12.69_65 _ Orn ui . 0 _':_ '" ,' ' , ; '" ; .... ; .... , ' ' ,- ' ' ' , .... ,-:-.'.". = .... , .... , .... , . . ,. :. ::-;Time ,_ o 200 400 600 800 10.00 12.00 14.00 1800 _ tt,J,o r,o,.< L,h ExacCt opyof 0riginal _ _/UZ_LL.o_---- Dat.e.... _......... ..................... ,'" _" _.-- 02048-21o7 solvent standard 10ppb 3M d021030013 o2o48-2s1o.7lvesnttanda1r0dppb 1 00 7 3M Environmental Lib d0210:_013 324 _.765) ,S,'n{Mn.2x 1.1_; Cm (322:326) dave0y707, 30-0_-2002 14:55:1S 1:Da_mrs o1513ES- 9.74 Davey070799 30-Oct-2002 14:55:16 2: MRM of 4 ChannelsES513 > 469 1.24e5 01 , , , i', , ,-, f ; , , ,5 ,-, -_, i ._ , ;i d0210300" 0/01 .i _+ 1=.3s4 3...9 IIl t 118.61o ..... .... , .... , .... +.... , .... ,-..-, .... i .... I .... i .... ,..-+..,., .... i .... , ,'T-tZ + 0 I"I DO 'too 15o 2oo 25O 3O0 350 40O d021030013 " 100 9.74 , ; , ,-_. ;-, . _ , , . : i , , ;-.-i . ,., , i 2: MRM of 4 Channels ES- ++1.68e5 2: MRM of 4 ChannelsES513 > 169 %-I d021030013 9.74 1.22e5 co 2: MRM of 4 Channels ES- i 513 > 119 _:1 1001 / 9.92 9.27e3 Or-- 8 13.65 =O _(_D 0 -- :'_ ; ..".. _ ' '"..":-;:;-. ...I.... " _ .... =......._..-._... ; - ,_ _: . . . , ,. , , _ , , . . , ,.:, . ,, . ,. = ; , Time _ o 2.00 " 4.00 6.00 8.00 10.00 12.00 14.00 16.00 _c_ t,J E)-< bo L/I ExactCopyof Original ,,' / _ / _._--_ IriitJal Oet_ +........... :---_ .................. :.... "_": ......... '+.......... S_ i : _'_,:_T:_ i golden west plasma 5x scale up 10123/02 ok 3M c1021030007 100- i i _ - ii !_ _i_ .o _ , _,: 10.34 Davey070799 30-Oct.2002 13:12:10 2: MRM of 4 ChannelsESTIC 1.13e5 i1%- 0.83 d020 1030' 0'07' I .... I_' 100t ] 10.14 _ 10.92 12.24 ]_ .... 1 "_ _ ,- _,=-u,,,_,0 ,,. _,_3E__"- 13.71"3.84 TIC 1.90e9 c_ "O__-_ o-L.--:-:-:...,..2..0_.0........, , . . . , .... , .... 4.00 6.00 ; .... 8.._00.... , .... _ .... 10.00 , .... j 12.00 - ._ -. 14.00 . ,..... . ; Time _ h.) .... hJ _,O-< ExacC_opyofOriginal plasma up Ig3doM0l2d1e0n30w00e7st 5x scale 10/23/02 ok 10.34 2: MRM of 4 ChannelsES30-Oct-20D0a2vey5106373:10>275:9119(9] 1001 4.37e4 II %] 0.73 0.1_'; -.J_. F. (1021030007 . i_-._. , i_', , .-i , _;-. 5.79 i ; .- ........ _ . ,=- ;-- ........ 10.14 11.30tl.98 12.1t712"6213.62 14.59 10.34 2: MRM of 4 ChannelsES563 > 219 1001 3.70e4, 9.18 10"181110"56 14.75 0 T, . .-, .... j .... , .... "i .... _', '' i .... i' " '-i .... i_.._. ,_"_l_..i.. J .... i-' .-" '-i "-, i ' "-"_' i : "-" ." d021030007 10.34 2: MRM of 4 ChannelsES563 > 169 1001 0.83cj'J-,u,="/"1.89 0_ ....... j_.__A. ,_.__.,............i..-...... d021030007 1"....... t" ,'....l...............l 10.14 2.93e4 , I 10.69 11.21 12.24 , '" '-r-'-"'_"p-_-' 10.34 13.40 '_';r; * .... ,,' ;. _'. ""-;- 2: MRM of 4 ChannelsES563 > 119 C_ _ :.; -o 1001 t " 3.33e3 O='Q %-] 7.76 10"18]1 11.50 12.0"4 13.17 ] _ 0.830."99990 _.. z]:> 3.57 _.4_ 6.44673 718 I 8438.69 _[ _, " J I" ,i 12..9'I..,_13,49 15.85 14.62 \./ r'_ 0= o0 o 0 __........__.._I _.... _ .... _I ".... 'l .............. , , , " ' ....; ' ; ............ , ; ,' ............. , , _-.,-._....'.".-_"w' Time '"__ _ '-_ 2.00 4.00 6.00 8.00 10.00 12.00 14.00 FO c_ _> .o_ O.< (Ji ExacCt opyof Original _ Date golden west plasma 5x scale up 10123/02 ok 3M Environmental Lab d021030007 320 (10.328) Sm (Mn,2xl.00); Cm(319:322) 169.073 1001 I 218.957 davey 07079S 30-Oct-2002 13:12:1[ 1: Daughtersof 563ES5.56e4 318.866 %- 96.702 168.29,_ \ 266.374 253.323 295.464 518.889 563.636 417.661 474.61g . 562.91l Co "; _ i_ O o 0 .... , .... , .... , .... ,".... , ...... _..,...'., .... , .... ,.;..,.'..,, .... ,'_.... ,...., .... , .... ,....,....,...., .... , ..... ,m/z _ '-_ 50 75 100 125 150 175 200 225 250 275 300 325 350 375 400 425 450 475 500 525 550 575 600 _ :0 t_J CO"< L,r= ExacCtopoyfOriginal In_ Da_ golden west 3M c1021030007 plasma 5x scale up 10123102 ok 1 00- golden _=t plasma 6x$_1eup 10/23/02ok 0021030007320(10328)Snt(Mtl.2xl.00);Crn(31=;322) 1130 1690"/3 dlvey 0707_ 1; DlUl_h__gl _83JES- 6.56e4 % =._7 10.34 10.14 \ Davey07079S 30-Oct.2002 13:12:1(; 2: MRM of 4 Channels ES563 > 519 0._ C d02103001 _. =.m, ="" _ 9.37 1..1,.30t.l...9..8..1.2.17 12.6,.2 1_3.,..8.2 14._5,9 _.. 2: MRM of 4 Channels ES- I | 100 I _'1r'_ _j_ _ 417.m| / ,!!,It" I ! ! 0 _*--,%': d021030007 ....i.N'-' ,_"' "_'""gO"'" g_'-'-'_'""i_=""-i,_'""=&'"_ "+:'i_ I .... I .... I .... "1 .... I''''' ' I .... 10.34 10.18 I .r. ; "_1 ' ' ' -'-'r -_" ".' _ 10.56 0.34 563 219 3.70e4 i '-'-* " '1 .... 14.75 i .... _'_-" "1 ; '+ " ' .. 2: MRM of 4 Channels ES- 563 169 1001 o+, 10.83 106 1.89 d021030007 ,,.:, ,.. 10.14 , .= _ t 0.34 2.93e4 v: 13_40 _ 2: MRM of 4 Channels ES- 563 119 o_ 100_ %1 (_ I 3.33e3 _"_ 7.76 10.1,,8 It 11.50 12.04 13.17 r" 3.57 6.44 7 18 8 438.69 ' 15.85 o 0_.... ,....,...., .... , .... , .... , .... , ....., .... , ....., .... , .... , ..... , .... ,....+.-... tl,,,e _ O 2.00 4.00 6.00 8.00 10.00 12.00 14.00 _ _) (O-< '-" ExactCopyof Original 02048-21-7 solvent standard 10ppb 3M d021030017 ,o1o_ | /o-_ I. .o ,o., Davey070799 I 30-0ct-2002 16:03:30 I 2: MRM of 4 ChannelsES- ( _-:'_1 O-m d021030017 100- I0.30 1: Daughtersof 563ESTIC 6.32e7 13.46 14_o4 O '-" _ __ _m_ o 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 --_-- (._ ::o tt,,o,.) _o-< (j-= /- Exact Copyof Original ,, _ i_ _ _+ i ....... F-i_ _%_+ +.:i_ ! _ i: i_:i _ _ _ 02048-21-7 solvent standard lOppb 3M Environmental Lab d021030017 319 (10.300) Sm (Mn. 2x1.00); 100- Cm (316:322) 269.007 - davey 0707991 30-0ct-2002 16:03:301 1: Daughters of 563E$- I 1.28e51 518.231 168.354 218.718 169.193 / 319.285 318.38E \ co ;! 98.859 (_ 1 369.374 .,.t.. 368.3 562.858 543 _o + ................ ,, , , , , I",',', _ .... , .... i .... _ _'_',, .'.', ,, .... ,m/z 0 50 75 100 125 150 175 200 225 250 275 300 325 350 375 400 425 450 475 500 525 550 575 600 _ ExacCtopyof Original _+._ ,,.,,,................. .,,........ ," ., ....,.,.+.............. .... 02048-21-7 3M (1021030017 100 solvent standard 10ppb %J 4 0 I, .. , , , . . ., .... d021030017 ', 100_ " _ ., .... -,...., %- ..... ., .... ,.....,..,.,.....,-_--_, d021030017 100 10.28 Davey070799 30-0ct-2002 16:03:3(] 2: MRM of 4 Channels ES- 563 > 519 2.68e5 ....... 10.28 -,.-. ._ .... ,.-...-, .... 2: MRM of 4 Channels ES- 563 > 219 1,77e5 _'_ _ ;-'I %-" "" I " " I .... i "'"" , I '" ' ' I 2: MRM of 4 Channels ES- 10.28 563 > 169 1.31e5 d021030017 10.31 2: MRM of 4 Channels ES- ;_ 563 > 119 " %-t] 5.02 6.80 8"1,,2,_8.37 9.05 12.24 12.95 15.24 om rO- 0 _ .... , .... i .... ., .... , .... , .... , .... "_-'...., .. '". , .... ,'..._"_--T-.,:.-_".. -,...- ..... ,Time _ 0 2.00 4.00 6.00 8.00 10.0( 12.00 14.00 16.00 ' "_ t,_ (D-K t-,J _J1 ExacCt opoy fOriginal /, t "LnJtlg Date "_ ......... "_" "- " ....... "......_. ..... -'-'-"......................... 02048-21-7 solvent standard 10ppb 3M d021030017 100 02041-21-7 solvesntat ndta0rpdpb d02t030017 319 (10._0) 8m (MIt. 2Xl.00); GI (318_22) 100+ 269.007 3M Environmental Lab "%t 0 ] .... I'' " d021030017 _ 21L718 1N.3S*. ] _"iTM dlVeOyT0.S t: Da_htlrl ol 553E._P 1.2Be5 5"18_2"31 30-.Oct-2002 16:03:30 10.20 Davey070799 30-Oct.2002 16:03:30 2: MRM of 4 Channels ES- 563> 519 2.68e5 i { ; ' ; ' "| + ' + + i-- i _ __ ' : ' 1 ' ' + ' | _' ' ' i i g':+ t "" ' "'L'' " " ' I 2: MRM of 4 Channels ES- ] ;'-' 100-1 /O t 90.11511 1 _1.314 ; 11 , _12.1151 l, 10,28 1.77e5 563 > 219 Ol d021030017 100-_ - _'"_'i'_'""i'_'":_'""_'""i_" "'-_'""_'""i_+'"'_ _"'_'g'""_ ''+ 10.28 2: MRM of 4 Channels E$563 > 169 1.31e5 %_ (Pq ,"+ Z "_ _-_ FO 0 -I" .... _ d021030017 "" ' _-+' ' + 'I : i :'I'' ' i .... I .... i .... i .... i ' ' ' '" I"'_ ' ' '_ "" '_ '" I'' ' ' ""_'' 10.31 , ' T ' ' ' ' i .... '= .... ; 2: MRM of 4 Channels ES- 563 > 119 1001 1.39e4 0 -.'1_ ' ' " ''I .... I " ' ' ' i .... 2.00 i .... 4.00 .5--.,0_2._ _ J .... I .... " 6.00 6.80 _ 8.?_ l '1 ' _'IIlIIl .... 8.00 'l 1'1' .._ _.__;24, ' : ' 'II'I ' II ' ' ' lII . 10.00 11 I ' ' I ' ' 12.00 12.95 , _';-T 14.00 15.24 ."J,/T-:'T-., Time 16.00 ExacCt opyof0rig+ha! ttm2_o-z._ LrdlJal [3ate _++_--.-..-.:.. ?,_P-.-........ . -.-_-- ----..-_-+~+_.-+.:...... l" ,-' v +..+ .._..... co rTl., :_ " _ _ O= _ _(:3 _O-< golden west plasma 5x scale up 10/23/02 ok 3M d021030004 100- 10.92 Davey070799 30-Oct-2002 12:20:37 2: MRM of 4 Channels ESTIC 4.55e4 i1%- 0.85 / 0.781 0.99 " _J_ 1"681"96 309 1_.47 0" " " " ' | " " ' " I ' ' " ' i '"" d021030004 3954.47 ' I ' " '% .645.546506.997"17.47 7.928.54 957 t0.75 11.6412i3012.47 , t._ 0.o4 9.57 lU.iO 13.12 I ' ' '' l " ' " i ' " ' ' I " "' " I " ' ' ' | " "' ' | " " " ' | "" " " I ' " " ' I ' " ' " ' ' " ' "I "" 1: Daughters of 613ES- 8.53 TiC 100 t 1.48e8 .it _'0 (_Q 02 I - . - _. [ 10 1210.43 12.84 tA "_') "_ 6.43 o o O_ 0 i, , , , ... ,,... , , .... , ... , .... , .... , .... , . .'.., .... , .... ",.... , .... , .... , .... ,Time _ 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 coco t_ o,.< ExacCt opyof 0dglnal _L_ _ _ [_ i_/_!J i " _i _ _i __-_ .... .C ! golden west plasma 5x scale up 10123/02ok 3M Environmental Lab d021030004 316 (10.910) Sm (Mn, 2x1.00); Cm (314:318) 100- 269.341 davey 070799 30-0ct-2002 12:20:37 1: Daughters of 613ES613.862 5.12e4 568.519 219.192 .. 295.037 319.180 360.130 418.715 CO _,, ;_" 209.292 "0 118.313 168.373 O_ 474.731 S77278 :_ c) 368.593 I 493.084 643.893 r-.- 0 o', _=o o ,-_ m/z t_ 50 100 150 200 250 300 350 400 450 500 550 600 650 __ Fo CO:o ExacCt opoyf Odgina] Init_ Date golden west plasma 5x scale up 10/23102ok 3M d021030004 100-1 - 4 %-I d021030004 100 10.92 10.92 Davey070799 30-Oct-2002 12:20:37 2: MRM of 4 Channels ES613 > 569 2.41e4 2: MRM of 4 Channels ES613 > 219 l 0.78 1.8_2 0 '. , , , i .... i ;_ d021030004 100 0.85 4.57 6f6 8.06 _ , ";--,=-;-=c,-. '---i ' -_ '"', ' "" , ' ' ' ' , ' 9._13 1023 10.50 11 43 ,-i .... = .... _'= '""' 10.92 1230 I .... 1254 I .... 15.09 15 50 8.59e3 a .... J ," . .; ." 2: MRM of 4 Channels ES- 613 > 169 1 I _.o9 % ...........9.9. , .... , .... , .... -,,...., .... ,.,..., .... , ........ ,.,..-, ,..,.. ,.-..., .-_,. _i: d021030004 11.33 2: MRM of 4 Channels ES- 613 > 119 .43 100-1 10.92 j 2.09e3 o_ %] 0 85 J 10.61 It lt115o c'-_" "t 058 _'"092 2.572.92 3.30 ._ 5.16 I _"_3 785 847 93"7 ",9,_"51 10.23\J iJil 1ill1'"(124 0_ 1l 2i.85.. _ 14.951529 15.85 _r" o--a m _0 =t ' 2.23 4.09 4.85 "." "" "1,.t.:'_0 .. O 0 ".... , .... , .... ,' ' ' ," '" ;' _' , .... ";" ' ' , ' ' '' ,"'' ",' ' , .... , .... _-'. , .-. , .... Time "_g t_ 2.00 4.00 6.00 8.00 10.00 12.00 14.00 ,_. ExacCt opyof Orig/na/ ,.' .l/:. golden west 3M d021030004 100- plasma 5x scale up 10/23102 ok llloldlm west plasma Ilx scala lip 10/23._2 Ok 3M Envllonme_ttal I"b d02103Q004 318 [10.910) Sm (Mn. 2xl.G0); Cm (314:818) 100- %1 O..J clayey 0707l!1 30-0t-2002 12:20:31 1: Daughters ot 613ES- et3.M2. 6.12e4 ==._lg " 10.92 Davey07079_ 30-Oct-2002 12:20:3"/ 2: MRM of 4 Channels ES613 > 569 2.41e4 96 . . _ ! -- d021030004 _k I00 _,=I._=z ' _t J .... I " " " I " ' i ' ' " "I " " I " " " ' I .... I "' ' ' 2: MRM of 4 Channels ES- 10.92 - 613 219 t I rl:=...,= ! oo,j IIIl, l ir-i ._ ............ 10.50 i "; "",___. _,_ ^ __K- o 0" ; , I .... I " ' ""i ' ' ; ; I .... i " ' '' I .... 1 .... '1 .... I .... I .... 1 .... I .... I .... I .... I ..... " d021030004 2: MRM of 4 Channels E0- IL 100 0.85 , 10.92 613 > 169 1 I'0_.99r"147 5s5 7.92 9.9_ _i: _: d021030004 100] " 11.33 10.921 2: MRM of 4 Channets ES- 613 119 . (b 2.o9e3 t 6.71 _ II.1.40 _' (rq %t 0.8=5 IJ 951 10.6_1, IIII 1I_111520" 12.85 t 058_("0"92,,_- 2"572"92 3:30.,,,, .6<_16 _713 7.85 8.479"37"1"_ 10;23_ II 11_12"41_. It 1336 14.951529 15.85 't-__ 2.00 4.00 6.00 8.00 10.00 12.00 14.00 bo L./t ExactCopyof Original _co;_o) o-.< 02048-21-7 solvent standard 10ppb 3M d021030014 100- I1%- 10.76 Davey07079S 30-Oct-2002 15:12:19 2: MRM of 4 ChannelsES- TJC 1.09e6 d0;21030014 100 10.75 1: Daughtersof 613ESTIC % 8.26e7 co 13.89 / 1:_ r-- _ Q '..0 0 a ' ' ' ' ,' '' ' , .... ',' ' ' "' I .... ,"' '"'-I .... I ' "' ' I .... ; .... ; .... , .... g . ,-'-. ,-. ' . . , .... , . .'.';Time ,_ O 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 ""_" FO co:_ t,O O.< ExactCopyo,"Of'i;ina! ,.. C.rlfql _)'/__---- " Inl1_ Dato 02048-21.7 solvent standard 10ppb 3M Environmental Lab d021030014 312 (10.754) Sm (Mn, 2xl.00); Cm (309:318) 100 318.703 Z2 !'. davey070799 30.Oct.2002 15:12:19 1: Daughters of 613ES- 569.293 1.27e5 568.459 \ %_ 169.029 268.267 216.536 .269.102 168.431 \ 368.951 o I-o .... 118.253 338.970 l 419.013 50 100 150 200 250 300 350 400 450 500 550 ' 600 650 _-_._C,otpyof Original :nitl_ Da_ __ ...... '_" ............... / ''"_"_ -"";"........... r_ 0 _.. _ _o:;o i _,"i 02048-21-7 solvent standard 10ppb 3M d021030014 100 d021030014 3, 100' %. d021030014 1001 d021030014 100 10.76 10.76 10.76 10.76 Davey0707991 30-Oct-2002 15:12:19 2: MRM of 4 Channels ES613 > 569 5.70e5 2: MRM of 4 Channels ES613 > 219 2.41e5 2: MRM of 4 Channels ESo 613 > 169 2.53e5 2: MRM of 4 Channels ES613 > 119 2.19e4 0 -T-:..-, .... , .... , .... , .... _-,_,, ; , .... . ,.. |.. ,;, ^,... _ ,,, ,_JL| ,,__ .... ,-:.-._'3-- .... ,4. :--,Time ,_ 0 2.00 4.00 6.00 6.00 10.00 12.00 14.00 16.00 _ t_ (o-< _ _<_cC_opyo_Original n_,_at D_te _ IJQ ,t_- _-,O t,O t,O 02048-21.7 solvent standard 10ppb 3M d021030014 100 0201s-=1-=7mwm=t=,da,=loppb 0021030014 312 (10.754) Sm (Mn. 2zl.00)_ Cm (30_):315) 100- 3M Envhonrnmnla! Lsb _=,._ _707_ 1: Daughter=of (513ES- 58J!.2g; 1.27e5 305-8O8.4t5-29002 lS._Z;I= | %t 0 j .... "'I d021030014 _ :18_o= 1,.0,I..=+ 118253 _+,, =m._1 ""I " ' ' ' l .... %" d07.1030014 100 ..... I I 4,.,I.m . 05_+_++_+++_+_+_++_+_++_+_+_+_++_5+_++ 10.76 I ' ' ' _ I + ''" ' I .... It 10.76 Davey070799 30-Oct.2002 15:12:19 2: MRM of4 Channesl ES- 613 569 5.70e5 | ' '"''t I'-I + ' ' I .... ! 2: MRM of4 ChannelsE$- 2.41e5 2: MRM of 4 Channels ES613 > 169 2.53e5 %1 0 +,,, I, I ''' l ' ' ,"r !, ' "' 1 .... d021030014 1100 % I .... 1''' ;I ''; '1"'7_i _ ' _' I ; ' +;" _ ' '' ' I .... 10.76 '1''' "' !' ' '= I ,'r'V , _ 2: MRM of 4 Channels E$- 613 > 11! 2.19e4 0 _,+-;_ '::i '-' '--' +_='--' ,' ..'....._ ,. 2.00 4.00 9.o7 : 4'_-._m-' , ;-,'_7' .3. 6'+_"++-.'= ,A_+'- ....,.... J l, "_ . ,+.'". +,- , -.--.','". .' + , .... , Time 6.00 " 8.00 10.00 12.00 14.00 16.00 / ExacCtopyof 0rigfna! ..' Lrdtlal Dat+ _, < +m ,_ _" (,O:_ _t3-< ._ golden west plasma 5x scale up 10123102 ok d021030008 3M .9.08 Davey070799 2: MRM of 3 Channels ES- 304_-2002 13:29:2T3IC 100II1% 8.811 1.12e7 t" L O , | i .... I .... i ', i , , i .... i , , , , i"',', , , ,-,", i , ,' , i'",' ! , , , t , , t , , , s , + i d021030008 1: Daughtersof 499ES- 9.10 TiC 1001 0_ 8._ 9.18 , 2.60e8 r_ t_ o 0 _ ..... ...., .... ,'. .... , .... , .... , , , ......... , .... , .... , , .... , .... ,Time "_ _ "_ 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 _ h,) (O'.< ExacCtopyofOriginal ln_d Date  i '_= i_: _ _ _ goldenwest plasma5x scaleup 1012310o2k 3M EnvironmentaLl ab d021030008 329 (8.884) Sm (Mn, 2xl.00); Crn (323:334) 100- 79.695 davey070799] 30-Oct-200213:29:23i 1: Daughters of 499ES1.67e6 129.614 % 0_ -P_"" _1'__3 98,714 "_: 179.580 229,786 168.663 280.108 "" "" 60 ': "r'l'" l""l'"']':' ;'"] _,i:],,,, _;'l I'" 'H'I'"'I''"P"'I"'- 80 1D0 12_ 140 160 180 200 220 2"']""I'-"'I _ 240 260 _'_'_'i;'_'_''''_';_i_'_''_''''i_'''_'_'_''''_;'_'_;;'_;''_'_'_'''_'_'_ 260 300 320 340 360 380 400 420 440 460 4_9';9'' .0'I7r_9'r___T;l r_z 480 500 520 ExacCt opoyfOriginal Initial Dete :o O r_ _" _ or_ cc__;0 golden west plasma 5x scale up 10123/02 ok 3M Environmental Lab 0021030008 337 (9.100)sm (Mn,2xl.00); Cm(334:343) 100- 79.695 davey 070799 30-Oct.2002 13:29:23 1: Daughtersof499ES499.1442.02e6 98.650 ,o % !l 129.614 r_ >_, f 168.663 229.979 - L LIII Jt_ ] ( '';'T"%-'_[ T .... I'" P'"I'"'I''"I''"I" 'l"nl='"l H" I"_'l'"'_l='"l'";'l ='" _''__'_'_'_'T'(_;'_7_%T_''_''__P_-;_''_.n_''_'_'''_''_'_''_'_ "== .;;l;..o1 "_ 60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500 520 O t',3 r,D '.< "_ ExacCt opoyfOriginal tn_ golden west plasma 5x scale up 10/23/02 ok 3M Environmental Lab d021030008 337 (9.100) Sm (Mn. _1.00); Cm (323:347) 79.695 100] ! I davey 070799 30-0ct-2002 13:29:23 1: Daughters of 499E_. 1.60e6 %- 98,650 499.144 129.614 = l 168.663.,,179.580 229.851 co 'l d _0 0 " _ "_ o 0 --" .. _L..J, ___-_L_ ..... _ ............. , h_'l,,"l,"'l,,,,I,,"t,,,'l'-'l'" '"" I' ......I.... I *I" .....I , I ..... I ' ....I. , I"" |,,H |,," I,',,| ........I........I........I I I I ' .....I n I"" I", ..I........I I "i ..I... i,,,; _;,11I'_" 1", ...I.. I,,," I""1 _,, ..i.. ,i .... I'n' I'" r'l m/z 60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500 520 _) to co;0 tJi ExactCopyof 0dgin_ ./!," golden west plasma 5x scale up 10123102ok 3M d021030008 1001 %-_ d021030008 1 100t Davey070799 30-Oct.2002 13:29:23 2: MRMof 3 ChannelsES- 8.81 499 > 130 2.45e6 9.08 | I I | I I 2: MRM of 3 ChannelsES- 499 > 99 4.19e6 o. d021030008 100 2: MRMof 3 ChannelsES- 9.11 499 > 80 ::0 I ":i. 0 6.75e6 '=" _ ;; ---.I 0 i , _ ; , , , , , , i , , i , Time __ 2,00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 _ ho C_ tt,j_O (O-< ExactCopyof Original /" . goldenwest plasma5xscaleup 1012310o2k 3M ! d021030008 1007 golden west plasma 6x scale up '10123/02 ok 3M nvilonmsntal Lab dsvly 0707St 30-0t-2002 13:29:23 8.8' 021030001L 337 lg.100JSm |Mn, 2xl.00]: Cm (323:347) YO.U6 I:Oaughlllirsof49gES1.50B8 I oj 1]oo] iI.NO 4aBB.I,_, o cl021030006 100 '_"' l,,,uo =='1"' ,Li 1011,t._! ,;I l! I _ 2eOi ...... o_ ":l_ ",_o z_ =;_,0--_o =_o" 4_"-;_=lo"--_O''== % 9.06 / .d _.., 9.08 8.89 Davey070799 30-Oct.2002 13:29:23 2: MRM of 3 Channels ES- 499 > 130 2.45e6 2: MRM of 3 Channels ES499 > 99 4.19e6 d021030008 100 % .._, ,-_ 2.00 4.00 6.00 t,_ Exact Copyof Orio!r__a .... . lail_a! Date 9.11 8.81il 8.00 10.00 12.00 2: MRM of 3 Channels ES- c_ 499 > 80 -_[: 14.00 Time 16.00 ._ _," r,n __ _ o,.< 02048-21.7 solvent standard 10ppb 3M d021030018 Davey070799 30-Oct-2002 16:20:35 2: MRM of 3 Channels ES- 9.11 TIC 6.02e5 0 10300 18 dO.2 8,81 9.10 1: Daughters of 499ESTIC /1 _ :t I o _ r..- _0 0 .... I .... I ' '"-""r'" . ' I '""'", .... ,"' "" I .... i .... I .... , .... i '_'"'w .... i ."" ", .... i . ''_ , ,Time i_ o 2.00 4.00 6.00 8.00 10,00 12.00 14.00 16.00 o'__) bo o-< h.) ExacCt opyofOriginat t r" Initial f)_l-_ ....... '.......... i._... i_ : _ _ _:_ _ ,_ !!_? _ii_ _ '; :_: =_ 02048-21.7 solvent standard lOppb 3M Environmental Lab dO21030018 329 (8.886) Srn (Mn, 2x1.00); Cm (321:334) 79.695 davey 070799 30-0ct-2002 16:20:35 1: Daughters of 499ES6.00e4 129.679 168.663 180.093 498.117 ._ _ ., 160.764 98.s86L_ =9.594 t,J O 60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500 520 _(_ tt__ (0-< i ExactCopy of OriQina_ -" ,p ,,/_L/U'L-- Initial Dai_ .... ..--.-...... _ ................ .__......_:,.;..,.:........ _.._.,... ,:__ ...... _._ i _-:Z-_ . -: , ...... , _,_-_-_;-- .; --i-_i ---_........... 02048-21-7 solvent standard lOppb 3M Environmental Lab d021030018 337 (9.102)Sm (Mn. 2x1.00);Cm (335:342) 100- 79.760 98.586 davey 070799 30-0ct-2002 16:20:35 1: Daughtersof499ES1.20e5 c_ 129.422 _;I 1 168.792 229.530 499.528 ! r-'- b_ 0 60 8D 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500___..520_ _ t,_ o-< F,.) ExactCopyofOriginal ,," g}L_Cl/__ lilt/_,n__. lailial Date _ ........... '_--_ " : .... -'-_""'-.... 02048-21-7 solvent standard 10ppb 3M Environmental Lab d021030018 337 (9.102) Sm (Mn,2xl.00); Cm (322:342) 100 79.695 davey 070799 30-Oct.2002 16:20:35 1: Daughtersof499ES8.55e4 98.585 04/0_ 129.679 168.663 180.093 _1 _"i 0 __ _. 498i17 :_ 0 .... I""l'"'l''",'"'l''"l'"'l""|'"'l""]'"'l""l'"'l''"l' ...'._._.h._..._....i._._..._.._..._|_._'_'_.'_._..._..._'...'_._.'_.._._.._._._...._ "'l''"l'"',""|'"'l'"'l'"'|''"l'"'|'"'|'"'l""l'"', r_z i1_ O 60 80 100 120 140 160 180 200 220 240 260 280 "300 320 340 360 380 400 420 440 460 480 500 520 _c_) _'_ " co::o t_o q3-_ L_ ExactCopyof 0r!::..: t" _c . cl,,'__.u./',--. Initial .r]_l_.. -._............,._........ _.._.._:-._................ - _.--,_-- _7,: : _ _ _:_ :_ _,_ _:_, _:!_ /_ ,_ _ .... 02048-21-7 solvent standard 10ppb 3M d021030018 100-1 l ] " ,I, , %-i d021030018 100 t 8.84 9.11 ! 9.11 Davey070799 30-Oct.2002 16:20:35 2: MRM of 3 Channels ES499 > 130 6.92e4 .- 2: MRM of 3 Channels ES499 > 99 2.23e5 %- 0 I .... _d021030018 100 I' , _ . , I ' , , 9 I .... I ' "' .'l'' ' ' ' I ' ' ''"* .... 8.87 \ I ' ' '/X _" ' I , ,', , I .... 9.11 I .... I _ , . I i :':"i , : i 2: MRM of 3 Channels ES- 499 > 80 o 8.81 I"" /1 .... , .... , .... ," ' ' '', .... * _ .... _ .... _ ' ' ' , . _ .., . .'.. , . . .-. , ."; _ . ,'. _.., .... ,Time .1_._ O 2.00 4.00 6.00 8.00 10.00 12.00' 14.00 !6.00 ---__ t_ ;o u_ ;,.,_yof0riglnaJ ,.'" :_._i_l _j_. ".,--....... _'._........ -_ .-z, ............ ..... ................. '2. i _ _ _ _:_ '_ _ _ _!.... _ _;_i;_ _ _j J 02048-21-7 solvent standard 10ppb 3M d021030018 100] | | | 02048-21-7 solvent s=nd;Ird _gppb 3M Environmentzl Lab d0210300t8 337 (9.tQ2) Sm (M_, 2x1.00); Cm(322:3_) , dJvl 0707_ 30-0ct-2002 16:20:35 1: Oaugll_ mol 490E_q- 8.84 Davey070799 30-Oct-2002 16:20:35 2: MRM of 3 Channels ES499 130 6.92e4 _ 0 t',J t,o " 9.11 100] %60 l==,_u_=s 100 .! ........!.1 _._K-i 150 200 250 0 300 3_ ..,I_ 411.117 400 450 500 , , ..... d021030018 100 , ..... , ,, , , , , , B_,_!.87 , , .... 1.11 % 8.81 499 > 99 2.23e5 , ,, ,o,, ., , , . ,, 2: MRM of 3 Channels ES499 80 3.46e5 ............. 2:00 4.00 ExacCt 0_yofO_igin. a_. tnlu_ Da_ .......... 6,00 8.00 10.00 12.00 /" . 14.00.............. , Time !6..00.- _"%i_ 0 _ 0 ,_ _ G,);o to-,< 3M ENVIRONMENTALLABORATORY E02-1039 i. I ATTACHMENT D: PREP SHEETSAND TRACEABILITYINFORMATION i t i ii ! ,i Page 125 of 225 3M E_IVIRONMENTALLABORATORY i E02-1039 ,I :: 1 : *, :__r i ." ':i a _ , ::j, , SPE Columns Extraction o:. Worksheet Prep Date: Analysts initials: Sample Number or description i Blank milli-q Lampire Serum Sigma Sem..m GoldenWest Serum Biomsource Serum Lamp.irePlasma 'GoldenWest Plasma Innovative Resourcets Plasma 10123/2002 OK /O ,_.-_ C__ _)/_ Method Revision: ETS-8-231.1 Study Number: E02-1039 Volume ot sample filtered (ml) Type of column Used 2000 ml 2000 ml 2000 ml 2000 ml 201711ml 2000 ml Waters, 19, 6ml Waters, 19, 6ml Waters, 19, 6ml Waters, 19, 6ml Waters, 1_, 6ml Waters, 19, 6ml I::luUon Amountand spike mix used . .. NA ...NA NA NA HA NA solventand volume Comments of MIOH TN ,.,al_ 2n1_M MIOH TN _13 2ml_ MIOH rCR._ea TN, _13 rcR.6a9 2ml d MeOH TN. A-6313 2ad _ MeOH _13 _ OfMeOH ,.=13 2n11of MeOH TCR-890 TN, rc_._7 TN TCR-a_ "IN 2000 ml Waters, 19, 6ml 2000 ml Waters, 1_, 6ml NA _Jofu.oN=T"N63m1R3.sa4 NA _ mR.sa3 ,,, "!_ '_ , ,, J '; Extraction=Steps(initialwhen/if done): ,;,._ _x_c0ndltion column with MeOH; x_wash with milIi-qwater;,_x_filter sample; ; ....!. _x_vacuum dry the column; _.x_elutecolumnwith _lvent; _'', % . ",. Additional comments: i 10mlofsera dilutedwith40ml ofmilll-qand200mlofACN.Sampleswer_ shakenfor20 minat 300rpm.thencentrifugedfor 10rain @ 3500rpm/4C Samplewas transferredinto1750mlof milll-qandfilteredthroughconditioneSdPE / :,}-,] ExactCopyof Original Date Page 126 of 225 , i 7:1 ! :I j "J l:l:!j _::3 ' i_ .i ; _-:_ _1 '! I ' i i _._ .; 3M E'/VVIRONMENTALLABORATORY E02-1039 Norf"GLP SPE Columns Extraction Worksheet Prep Date: Analysts initials: _}_ 1011112002 'c. la.o'_ OK OL/--J Method Revision: E-T-S..;_Q-I _) Study Number:.E 02-1039 Sample Number description 'H _ 31ank,milli-q-1 Blank milll-q-2 rabbit serum blank rabbit serum curve-0.1ppb rabbitserum curve-0.25ppb Volume of Elution or sample Type of column Amount andspike solvent and filtered (ml) used mix used volume Comments '" 2 ml'of MeOH 400 ml Waters, 19, 6ml NA TN-A-=10 2ml ofMeOH 400.ml Waters, 1_],6ml NA r.-^_lo I ml_ MeOH 400 ml Waters, lcj, 6m! NA rN_._o TCR.See | mlof MeOH 400 ml Waters, 1_, 6ml 2ul of 02050-14 rN-A-aam rCR._8e 400 ml Waters, 1,cj.6ml 5ul of 02050-14 rN.*-=_o ! ndol MIOH rCR-aee rabbit serum curve_-0.5ppb irabbit serum curve-O,.75pFb ,. rabbit serum curve:lppb I rabbit serum curve-2.5ppb i I rabbitserum curve-Sppb . : rabbitserum curve-.10ppb ;.._ '" rabbit serum curve-_30ppb Lampire serum blank-1 Lampire serum blank-2 Lampire serum MS K)..5ppb Lampire serum MS-Sppb 400 ml 400 ml 400 ml 400 ml 400 ml 400 ml 400 ml 400 ml 400 ml 400 ml 400 ml Waters, 1_],6ml Waters, 1j_6ml Waters, lg, 6ml Waters, 19, 6ml Waters, 19 6ml Waters, 1_, 6ml Waters, 19, 6ml Waters, lcj, 6ml Waters,,.lg, 6ml Waters, 1_ 6ml Waters,lg, 6ml 10ulof 02050-14 rI n_d-oaf 3MrIOoH 15ul of 02050-14 r_a3m 2 nd,_ MAVDH 20ul of 02050-14 50ul of 02050-14 T_a_o 2 mlof MIOH TN-A_tO 2 mlof MIOH 100ul of 02050-14 _A-ealo 2 mlot MeOH 200ul of 02050-14T,-A_o 2 mlof MeCH 600ul of 02050-14 r.-_.3_o 2ml of MIOH NA rN-_-_m 2ml _' kteOH NA rN-A-_m Z ml,V MeOH lOul of 02050-14 r,-*.-._o | mlol MeOH 100ul0f.02050-14 rN.A._o rcRaes rcR-aSe rcR.eaa rcR.ses TCR-_88 TCR.680 TCR-888 TCR.SSa TCR.SaB TCR._a TCR.eaa Sigma serum blank-1 Sigma serum blank-2 400 ml 400 ml Waters, 1_16, ml We!ere, !g, 6ml NA rtrr.n.l,o_f aMeaO,Ho TCr_S 12mlof MeOH NA TN-_-_I0 rcR.ea9 , t Sigma serum MS-0.S ppb " Sigma serum MS-5ppb Gold.enWest serum blank-1 400 ml 400 ml 400 ml Waters, lcj, 6rot Waters, 1_, 6ml Waters, lcjt 6ml 'TN-A-11O 10ulO__r,_o_. 22mml_f|Moel_MH eOH 100ulo2_-_hTN-_-e=_O 2 rnl _ _lldOH rCR-68g rca.ae_ NA _.._o rCR_O Golden West serum blank-2 GoldenWest serum MS-0.Sppb GoldenWest serum MS-5ppb ",. Bioreseource research serum blank-1 Bioreseource research serum b,L lank-2 Bioreseource research serum,MS-0:5ppb Bioreseource research serum MS-5ppb 400 ml 400 ml 400 ml 400 ml 400 ml 400 ml 400 ml Waters, 1_16, ml Waters, 1_ 6ml .waters, 19_6ml Waters, lg, 6ml .Waters,19, 6m!. Waters, lg, 6rnl Waters, lg, 6ml NA "rN-A-_m 22 mmllOoI t kM_eOOHH lOul of 02050-14 T,-_a_o 100ul of 02050-14 =_m.^lo_o_U,OH ! mt ol Me,OH NA TN-._-6310 2 rill OfMeOH NA TN-A" -6310 2 mlO| MeOH 10ul of 02050-14 T.-^-_=_O 2 ml Of MIOH 100ulof 02050-14 !Ta-x-ea_o rCR._gO rcR-ego ,IrCR.e e TCR..887* TCR-ea7 TCR-687 TCR.887 ii_ Extraction Steps (initialwhen/if done): '" _x condition column with MeOH; _.x_washwith milli-qwater; _x_filter sample; _xvacuum dry the column; _x._elute column with solvent; _" :" Additionalcomments: hC'_ _ .A.-_/_C_ c'_ ,c,,._,? ExactCepyofOr_ginal _ _(b"t_._ InYdaJ Data 2ml of seraJplasma diluted with Bml of mil[i-q and 4Qml of ACN. Samples were shaken for 20 rain at 300rpm, then centrifuged for 10 rain @ 3500rpm/4C Sample was transferred into 350ml of milll,q and filtered through conditioned SPE _,-,_"_" v_- _ C:c7% ,-',_;_'-"='-_'_\_'_H "_ _._c'_'_ _.;_;/a'Y.YJ- Page 127 of 225 3M E_IVIRONMENTALLABORATORY E02-1039 !i' i ,_:,.._ i ' i:- I : . "' ::::i , ! I :_; .- ; i .. !__h i'il_ ; 2:_} ' Prep Date: Analysts initials: ./ ._LP SPE Columns Extraction Worksheet pO__.l 'JP'10/10/2002 --/'" OK It.;/6'.o,_' C,p__. Method Revision:SPE validation Study Number:,E 02-1039 '" Sample Number ,, description Blank milU-q-1... Volume ot Elution or sample Type of column Amount and spike solvent and! filtered (ml) ii m used mix used volume 2ml ol MeON 400 ml . Waters, 19, 6ml NA m-A._lo Comments Blank milli-q-2 Chinese plasma curve-0.1ppb C,hinese plasma curve-0.25ppb. Chinese plasma curve-0.Sppb ' ' '" Chinese plasma curve-0.75ppb Chinese plasma curve, -lppb ! I Chinese plasma curve-2.Sppb i Chinese plasma curve-Sppb , ] Chinese plasma curve-10ppb _I, Chinese plasma curve-30ppb, Chinese plasma blank-1 Chinese plasma blank-2 Chinese plasma .MS-0.Sppb ,., i C,hineseplasma MS-Sppb 400 ml IWaters, 19, 6ml NA 2rNml-o_f 1M0IIOH Zml cffMeOH 400 ml 400 ml 400 ml Waters, 19, 6ml Waters, 19, 6ml Waters, 19, 6ml 2ul of 02050-14 5ul of 02050-14 10ulof 02050-14 rN-A.6310 ZmlotMeOH rN-A._0 2ml oIMeQH r_,.A.=_0 ImlolMeOH TCR-674 rCR-_74 TCR-(174 400 ml 400 ml Waters 1_18, .ml Waters, 19, 6ml 15ulof 02050-14 20ul of 02050-14 r....,,_rQ 2 ml of MeOH _-A-._a 2 ml OtMIOH rcR-e74 TCR-6"/4 400 ml 400 ml Waters, lg, 6ml Waters, 19, 5ml 50ul of 02050.14 _-,_._t0 2rid _ MIOH 100ulof 02050-14 :..A-ela 2 mlo( MIQH TCR-874 TCR-e74 400 ml Waters, 19, 6ml 200ul of 02050-14 m.A_31o TCR.874 2nd oiMeOH 400 ml Waters, 19, 6ml 600ul of 02050-14 m-A._o 2ml el MIOH TCR-874 400 ml Waters, lg, 6ml NA r..A.=_o TCR-874 Z mlofMlOH 400 ml Waters, lcj, 6ml NA rN_._to /'CR.674 Z ndot MeOH 400 ml Waters, 19, 6ml 10ul of 02050-14 m.A-=_0 l ml of MeOH rcR.e74 .... 400 ml _Waters,lcj, 6ml 100ulof 02050-14 _A._',o TCR-674 Lampire Plasma blank-1 ' i Lampire Plasma blank-2 ! Lampire plasma MS-0.Sppb _ Lampire plasma MS-Sppb I Golden West Plasma blank-1 400 ml 400 ml 400 ml 400 ml 400 rnl !Water , 19, 6ml Waters, lcj, 6ml Waters, 1_, 6ml Waters, 1_, 6ml Waters, l_h 6ml NA NA 10ul 100ul NA rN-A-_m t ml of MeOH m-A.=to | ml ofMIOH rN-A-=,o N..,'.-_0 _ml of MeOH rN_tO TCR-e_ rCR-68S rCR-88S rcR-eas rcR-ell4 GoldenWest Plasma blank-2 Golden West Plasma MS-O.Sppb Golden West Plasma MS-Sppb. 400 ml 400 ml 400 ml Waters, lcj, 6ml Waters, lg, 6ml Waters, 19, 6ml NA 10ul of 02050-14 100ulof02050-14 IrnNdo.(_MmeOoH _nd_ M_OH rN-A-(_tO _ml _ MIOH N..-_lo _ed _ MeOH rcR.ea4 "rCR-M4 rCR._ InnovativeResearch Plasma b!.ank-1 .... t Innovative Research Plasma blank-2 i 400 ml 400 ml Waters, 19, 6ml Waters, 19, 6ml NA r...._o rcR-s_ t ml of MeOH NA TN.A..6.110 TCR-eB3 r2 ml _ MIIOH i Innovative Research Plasma MS-0.Sppb_ 400 ml Waters, 19, 6ml 10ulof 02050-14 _'N_'^_"_.OO._ rcR-_a3 ' InnovativeResearch Plasma MS-5ppb 400 ml Water,s,,lg, 6ml 100ulof 02050-14 TN-_,H$310 TCR-683 ,,, Extraction Steps (initialwhen/if done}: _xcondition columnwith MeOH; _x_wash with milli-qwater;, _x_filter sample;; _x_vacuum dry the column; _x._elutecolumn with solvent; ExacCt opyof Original _.iY1C /O/_( ](_r-2..... A('d,:_d.i-tpio.n_a=l_comme,_n,t's_:-'_ C' 2 I1_-_,$_;_,'- , ,;_ _ _.._: V_j,_ _(_. (__,_ C_/(.:C_. 2mlofsera/plasmdailutedwith8mlof milli-qand40mlofACNS. amplewsareshakenfor20minat 300rpmth, encentrifugefodr10rain@ 3500rpm/4C Samplewastransferreidnlo350mlofmillJ-qandfilteretdhrougchonditionSePdE k'_ _A "_/_t Page 128 of 225 Y'-I : .... .1 .c, "] i !I ii _I i 1 i 3M ENVIRONMENTALLABORATORY E02-1039 3M ENVIRONMENTAL LABORATORY Note to File Project or Study Number: E02-1039 Associated Study Number: Forsamplesthat werepreppedon 10111/02t,hat areBioresourceResearchSerumMS 0.Sppband ._PP.,O_m_ .o_Cmw.-Pa_.]_Lnw_r_em._.aeem__n!'y, .sw_n_P,.......................... ..................................................................................... .................................................... ..................................................................................... ................... ..... i ?.i'i_ , i_I!i_ ', _Y'_ :i 'i .! , ', ,, 2'i ............................................................. ..................... .................... t ........................................................................._.....;.'_'_,,/,_: _ ":'%_ -_ - ................................................................................................................................ -_" Recorded By: I ,. / [ FormETS4-15,0 Page 129 of 225 i :_:?' i.,,,_ ....; ;:"i ::.,.ii .=i i :=, JM t-NVIH_JNMEN IAL LAIdUHA I UH Y E02-1039 Test Control and Reference Substance Log I Substance trade name IHuman plasma from China or reference # IITCR Substance # TCR-674 ISnaumbset:ance/chemicaI ILotJbatch#: 13M# (Human p,asma IN087P27 I ITCR# IReceived from: TCR-674 Northwest Bioanalytlcal Expiration date: I1 Avoml)o: unt received (wt. or 7x-lOml IlnitJals: IOK Date: 0912512002 Number/size of containers: I17-15 ml centrifuge tubes Shipper: FedEx ]Condition: Iliquid, Biohazard MSDS (y/n) '=..)Y N IRetain I Date of Retain 1.7.,_.'.-.-.......i[.._ .-_U_.:...-_-.-.._.-.-._;.__.--T_::...--..',.,:_--_.-._!_..,:_.=__-.:'; i(..: ArchwedlSubstancNeot:AVatlaq!e_i] IPumy: I I IRecor_s received: I I fabrication, or derivaUon Lreoccoartdiosn: of synthesis, I I IStdLocaUon/Storage: IFrozen F19 I IMolecular Formula: I I IComments IAttachment(s) ) Smearrekecdamoen Itnhe7 tsuebpear(a4t,e5,6tu,7b,8e,s1.1 Poler a1s2e) mark which tube was used for sample, bY number I I I Exact Copy of Original .... I_al Date- i : t Page 130 of 225 ,:. 3M ENVIRONMENTALLABORATORY , E02-1039 _,:_ "'__ _uman plasma from China "lumen plasma "CR-674 USE LOG Gross Wt.Nol. Amnt. Gross wt/vol Purpose (enter standard number Balance Balance Initials Date ,:,! Before withdrawn after or reason for removal) ID ID .. I' withdrawal (mass or vol) withdrawal Balance ID 1 Balance ID 2 Balance ID 1 1 2 . I'" 26ml " IcEo0m2-b1t0n3e9d, )validation (all tubes were _a _a IOK 1010912002 ! ExactCopyof Odginal I_al Data !:'23 Page 131 of 225 i b _"_,_i :<, .... ' _'; : =_. i ? '.:!_ 3M ENVIRONMENTALLABORATORY E02-1039 Test Controland ReferenceSubstanceLog Substancetrade name IPooled HumanPlasma In Sodium 3r reference# SCitrate TCR Substance# IITCR-683 1 I [nSaumbes:tance/chemical [Pooledhumanplasma Lot/batch#: IIR02-014 :3M# I Expirationdate: 09130/2007 nitials: IOK TCR# ITCR-683 [ Receivedfrom: IlnnovativeResearch,MI I I Amountreceived(wLor [10xl0ml vol): i [ Date: I10/02/2002 I Ncounmtabienre/srisz:eof I10xl0ml plastictubes Shipper: I I Condition: R-etain ]good I MSDS(y/n) t .) Y N I Dateof Retain [ I ! C')ArchivediSubstanCeNotAvailable:t IPurity: J I [Recordsreceived: I CofA I {fabdcatlono, r dedvation [iLreoccoartdiosn:of synthesis. ISouthfleldM.I I [StdLocation/Storage: I Frozen, F24 I lMolecularFormula: I I JComments I I A[Attachment(s), I I ExacCt opyofOriginal _C_ _,/, / o__=.= 'l .... 1 Page 132 of 225 3M ENVIRONMENTALLABORATORY E02-1039 I _ooled Human Plasma =ooled humanplasma _" i rCR-683 USE LOG I 1| '- _ I Before withdrawn after or reasonfor removal) ! I withdrawal (massor vol) withdrawal Balance(D 1 BalanceID 2 BalanceIO 1 I', 18ml I" IE02.103va9lidation I" I" IoK , 110114/2002 :i,.i ,i ,i ExacCt opyofOriginal Page 133 of 225 '.'"_:_ ,; :_ t:5',i .... i .,;.. 3M ENVIRONMENTAL LABORATORY E02-1039 I pooled Human Plasma Fooled human plasma _CR-683 USE LOG I Before I withdrawal ,IGBroalsasnWcet.NIDol.1 I' withdrawn (mass or vol) I BalaAnmcne,.ID 2 110ml after withdrawal t0B=-a-lIance ID 1 i" or reason for removal) Purpose (enter standard number lE02-1039v,alidation ID IBalco2JBa,aI =l,nOa,e,,a,s. l" b IOK 110/=3/=00= _ ExactCopyof Original I_"_ C/"/'1C tilt ,/ 0% Date J__ ! Page 134 of 225 , _"!:i.. I, i , .i i: :_""!i .." i _L-'.! ii ! 3M ENVIRONMENTALLABORATORY E02-1039 Test Control and Reference Substance Log I Substancetradename Sodium Citrate PooledHuman TCR Substance# TCR-684 J or reference# P asma ILoUbatch#: (3M # I ,ra,on Ilnitials: tG014t0002 I r,o,o3, o7 IOK Number/sizeof containers: Jlxt O0mlplasticbottle j ICondition: IRetain JFrozen I ;!( ).,. -Archive"d" "l.S" "u.'Cbas. ta: n' -c.'!e:N" 'ot.A .V" aLi:.l_a...:.b... le'iI IReceivedfrom: I Golden West Biologlcals, Inc. I I ovo)u: ,',,rec(ewoi,,r."e,00,',',, I IDate: 10103/2002 ] I tShipper: UPS i J IMSD(Sy/n) l" ) Y N J IDateof Retain I I (Recordsreceived: I CofA I Jfabdcationo, r derivation IJLreccaotridns:f synthesis' I Glden west Bilgicals I {StdLocation/Storage: (Frozen F19 ] (MolecularFormula: I I IComments J I }Attachment(s) I ] Exact _i CopyofOriginal C_rvl( _ jt/L(_._. __t _ Date (':1 Page 135 of 225 3M L_NVIRONMENTALLABORATORY , E02-1039 5odium Citrate Pooled Human Plasma ...i._, ;_ooledhuman plasma .i "CR-684 USE LOG i I Before withdrawn after or reasonfor removal) ID jBa,a,JBal cej l,o,,,a,s, I withdrawal (massorvol) withdrawal 1 .. I[GBraolsasnWcet/JDVo1l.I BalAanmcnetI.D 2 IGBraolsasnwcet/IvDo1lJ Purpose(enterstandardnumber o=e I, .. 18ml I" IE02-1039,validation I,a _a IOK 110110/20OZ ExactCopyof 0dginal _'t _ ' Initial Date Page 136 of 225 ,,. 3M ENVIRONMENTALLABORA:'I'ORY E02- f 039 :i I USE,OG I : I; _ooled humanplasma ' p[i'oCdRiu-6m84Citrate Pooled Human Plasma 1 ...i,o. oi.,._w,_-.-vr-os-o, = I 0..I ::! Before withdrawn after or reasonforremoval) withdrawal (mass or vol) withdrawal Ba anceIO 1 BalanceID 2 BalanceID 1 I- I_oml I" IE02"1039 I"A INA IOK I101=:_=00lZ ii:t _:;::I ExactCopyof Original. Date i_._'_. Page 137 of 225 ! ._ . '-! i ,,,_ J > " !' : 3M ENVIRONMENTAL LABORATORY E02-1039 Test Controland ReferenceSubstanceLog Substance trade name or reference # Substance/chemical name: Lot/batch #: 3M# Expiration date: Initials: Number/size of ;ontainers: Condition: Pooled Human Plasma From Lampire IPooled Human ILamp|re 122-60824A I II10108/2007 {OK 11-250mplalstic I Plasma bottle From jFrozen Retain 1 , IITCR Substance # TCR-685 ITCR # I JReceived from: I Amount received (wt. or vo ): TCR-685 Lampire 10Oml IDate: 101t012002 JShipper: I IMSDS(y/n) UPS _.) Y O N IDate of Retain I Punly: I I Records received: I Disease Screen I fabrication, or derivation Lreoccoardtiso:n of synthesis, I Lampire I Std Location/Storage: I Frozen F19 I Molecular Formula: I I Comments I I Attachment(s) I I ExacCt opyofOriginal _ Datll Page 138 of 225 : ....i.. :_:_:'! T ii _i:!!J 3M ENVIRONMENTAL LABORATORY E02-I039 _ooled HumanPlasmaFromLamplra _ooledHumanPlasmaFromLamplre USE LOG I I Before withdrawn after I withdrawal (mass orvol) withdrawal I BalanceID 1 BalanceID 2 BalanceID 1 orreasonforremoval) 1" 11Oral I" E02-10"19 I I I ID 2 Ina ha ' IOK 11023/Z002, _i1 ExacCt opyof 0dg_nal _ C("v"/(: i _/_/o-z-.- Oat= i, Page 139 of 225 3M ENVIRONMENTAL LABORATORY E02-1039 I USELOG I I pooled Human Plasma From Lampire I i_'_I _oolad Human Plasma From Lamplre i',_' _CR-685 _--! Before withdrawn after or reasonforremoval) ID withdrawal (massorvol) withdrawal 2 . GBraolsasnWcet.NDo1l. I BalAanmcnetI.D 2 BGarolasnscwetI/Dvo1l I Purpose(enterstandardnumber IBal_ce I BalanceI IniUals I Date I" 18ml I" IE02-1039,validation Ina _na IQK 11011012002 ei _;,_ i _it ExacCt opyof0rigfnal ] L !Li Page 140 of 225 ....,_. .... :L : " "= ::! " 'i '.:.'.._. "i = ,.o 3M ENVIRONMENTALLABORATORY E02-1039 Test Control and Reference Substance Log JIoSrurebfsetraenncceter#adename name: ILot/batch#: {J3ME#,p,,Ua,oen: I Ilnitials: PBoloorleesdouHrucme anSerum From roo,...oo. .ru 1020821 I I JOK ITCR Substance, i.c . IReceivedfrom: ITCR-687 IBloresource IAmo, ueniv,e,oI,OOm, VO): IDate: 110/11/2002 Ncounmtabienre/srisz:eof 15O0mpl tasticbottle IConditlon: JFrozen {Retain I ! ... , :.. ..... ... ... _. ' ,"" ,:' - " : L_--"-';:i "-_::'; ":%.',;_:J_: _( ,. ) Ar.ch. iv:edlSubstancb N0i_ AV. aii.a,ble_ :. IShipper: IMSDS (y/n) IDateofRetain UPS 1..J. Y N I IPun_/: 1 :" I lRecordsreceived: I I Ifabricallon,or derivation [ILreoccoartdiosn:of synthesis, 1Sioresource I IStd Location/Storage: I FrozenF19 I IMolecularFormula: I I IComments JAlso assignedTN.A-62841011110O2 K I IAttachment(s) I I r", (_ ExaCtCopyof Original 'nEz.Vj._.___e._ !_J Page 141 of 225 3M ENVIRONMENTAL LABORATORY E02-1039 I ooled Human Serum From Bloresource _!_i _>ooled Human Serum USE LOG BaracI,.'o'IIvG",w="0w'.'B,"u.v",n.=o'orOn,,.IoVO,I",'"/'Or"Ia0"" 11Oral I" IE02"1039 F J" I0K' 110/23/2002 Exact Copy of Original _,W(1 I,,/,/0_ Page 142 of 225 3M ENVIRONMENTAL LABORATORY E02-i039 i I ooled Human Serum From Bioresource _,. Fooled Human Serum ;_,I FCR.687 USELOG I Before I withdrawn after or reason for removal) ":i I withdrawal I (mass or vol) withdrawal I Balance ID 1 I Balance ID 2 Balance ID 1 [- Is'" I" E02.1039, validation I" l" IOK 110111/2002 ! ,t IL. ExacCt opoyfOriginal _ Date ; Page 143 of 225 3M ENVIRONMENTAL LABORATORY E02-1039 _' i I Test Control and Reference Substance Log ':""_ or reference # , ! Substance t,ade name IPooled Human serum from Lampiro TCR Substance, TCR-.688 I I i, _I Substance/chemical jPooled Human serum TCR # TCR-688 name: JI 3M# I Lot/batch #: IX324B " I I Received from: ILampire J Expiration date: (0812912007 I Avoml)o: unt received (wt. or 1, 250ml I Initials: IOK Date: It 0/11/2002 containers:NUomf ber/size ,JZSOml Nalgene bottle Shipper: UPS I Condition: JFrozen USDS (y/n) _. } Y t_ N I Retain J Date of Retain I [ ' ( 5"ArchivedlSubstan_eNotA_iia_,e_ ::") Purity: J I Records received: J J I I ' i Location of synthesis, I Lampire J t fabrication, or derivation '-.,; records: Std Location/Storage: JFrozen F19 J i Molecular Formula: J I , Comments JAlso assinged TN-A-6286 10111102ok I Attachment(s) J I _i,:!i: ii_ii 'i ExactCopyof Origiac,l J Page 144 of 225 ; i:i 1 )ooled Human serum from Lempira i _)ooled Human serum ,; I_'CR-6B8 3M ENVIRONMENTAL LABORATORY E02-1039 USE LOG -i I Before withdrawn after or reason for removal) ID : _ withdrawal (mass or vol) withdrawal 1 I GBroalsasnWcet.NIDol1. I BalaAnmcenLID 2 t BGarolasnscewtI/Dvo1l Purpose (enter standard number Balance I Bali)nce I IniUals I Data I" 18ml i" E02-1039, validation I" _" IOK 11011112002 'S= i !, . J i J !i'? _ i ,,ii ii o Exact Copy of OHglnal i Page 145 of 225 3M ENVIRONMENTALLABORATORY ..... E02-1039 "'i I USELOG I I pooled Human serum from Lamplre [ _ '_ pooled Human serum 'i Ircr.68e :" i Before withdrawn after or reasonforremoval) ,t withdrawal (massorvol) withdrawal Balance D 1 BalanceID 2 BalanceID 1 l" 1O1 ral I" [E020-:t19 I" I" ]oK 11012_:z00:'I i :i :1 I i_ Exact Copyof Original !",'L 1 Page 146 of 225 i ,,._ --,_ _'_ .' ._ ' '_! ::': ! '_ !;.:"; :_":r_: -::_, ,, :' '_ ; 3M ENVIRONMENTAL LABORATORY EO2-'103g Test Control and Reference Substance Log I Substance trade name JMale Human Serum from Sigma or reference # IITCRSubstance # TCR-689 Substarlce/chemJca, name: Lot/batch #: 3M # Expiration date: Initials: Number/size of containers: Condition: Ii_t_ale Human serum 1022K0965 I I10712412007 IOK 1-100ml plastic bottle Ifrozen J'l'CR, I JReceivedfrom: I Amount received (wt. or vo): IData: [Shipper:. I IMSDS (y/n] TCR.689 Sigma 100ml 10/11/2002 UPS _Y N _Rem!n................................!_._. IDate of Retain i !-; ArchwedlSubstance Not Avadab] e '1 !Purity: I Records received: ] ...... ] ] fabrication, derivation Lreoccoartdiosn: of synthesiS,or I Sigma I Std Location/Storage: I Frozen F19 I Molecular Formula: I I Comments I AIso assigned TN-A-6250 10/11102 OK ] Attachment(s) I I ExactCu;;,.:.:,?_,.. ,,; :i?i Page 147 of 225 3M ENVIRONMENTALLABORATORY E02-1039 I USELOG 1 / _AaleHuman Serum from 51gma ] ' i"ii pale Human serum ! irCR.6a9 .... '/lJBGwarilBtashensWdfcoretar'INewDal1"l I (BmwaaliAatshnmsdconreraLIvDwon2l) BGwariltasahnsfdtcwereratI/Dwva1ll I Purporsree(eanstoenrfsotrarnedmaordvnaul,mber IBal_ce I Bajl2IaDnceI Initials. Date I I- le_nl I" 1E02-10v3a9n, datlon I" J" IOK 110/111202 I j q _,,_ Ng!,', !i . ,! ., ! ExacCt opyofOriginal Page 148 of 225 1 I ") I pale Human Serum from Sigma '__i pale Human serum .i, FCR.e89 3M ENVIRONMENTALLABORATORY E02-1039 USE LOG / ;i I Before withdrawn after or reasonforremoval) ' I withdrawal (massor vol) withdrawal I BalanceID 1 BalanceID 2 BalanceID 1 [" [lOml I" [E02-10,19 PF IOK [10123/Z002 l i_,':,=I ,] r) "y_-4 '_";"_i i ! t:;.:!] ExacCt opoyfOriginal Page 149 of 225 i _i "i ;:;"I 'h"l: : ; -' 'i " , 3M ENVIRONMENTALLABORATORY E02-1039 TestControal ndReferencSe ubstanceLog Substancetrade name 3r reference# Substance/chemical aame: Lot/batch #: 3M # Expirationdate: Pooled HumanSerumfrom Golen ,West Pooled Human Serum G01406042 0712512007 Initials: Number/sizeof containers: Condition: Retain OK 1-500ml blastic bottle frozen TCR Substance# TCR.690 TCR # Receivedfrom: TCR-690 Goldenwest Amounrteceived(wt.or 500ml _ol): Date: 1011112002 Shipper: UPS VISDS(y/n) Date of Retain _.) Y N i !:..(.".'- Archived/Sbbsta, nCe.N.oi.;Av_iia..l._[ie::.,.: .;: Pu_/: I I Recordsreceived: I I I I Locationof synthesis, I Golden West I fabricationo, r derivation records: Std Location/Storage: IFrozenF19 I MolecularFormula: I I Comments I AlsoassignedTN-A-62511011110o2k I Attachment(s) I I !;_;;_ I ExacCt opoyfOriginal _ Dtlto Page 150 of 225 3M ENVIRONMENTALLABORATORY i E02-'1039 ! ' I : '_i p_o'ooolleeddHuman Serum from GolenWest _rCR-690 USE LOG -:" Before withdrawn after or reasonforremoval) withdrawal (massor vol) withdrawal GBraosasnWcet.DNo1l. I BalAanmcnetI.D 2 I BGarloasnscwetI/Dvo1l Purpose(enterstandardnumber I Bal_ce I Bal_)nceI Initials I Date I" 1lml I" E02-1039 l" F IOK I10/2312002 z r:7 li_i__, !i:iJ i:cli! L _ i ExacCt opYofOriginal" Page 151 of 225 i? _.i :'_"'i,,, ,, i : _:i 3M ENVIRONMENTALALBORATORY E02-I039 I USE LOG I I _ooled Human Serum from Golen West I _PooledHuman Serum _rCR-690 Before withdrawal GBraoasnscWe t.NDol1. [" withdrawn (mass or vol) I BalaAnmcenltD. 2 [8ml after withdrawal J BGarloasnscewlDt/vo1a l" or reason for removal) Purpose (enter standard number E02-1039, validation f Bal_ce [ Bal_nce I InlUals I Date [" _" IOK 11011112002 ._:._,_ ExacCt opyofOriginal Page 152 of 225 Standardcurve trackingsheet P,ep date: Analyst: 10/7/2002 OK /(.._'C_t/ ] 'C-./"%J Type ofthe cun_e: Target analytes: Solventcuwe forSPE sera validation C6, C7, C8, C9, C10, C11, C12, THPFOS,THPFDS, PFOS Standard mix usedforpreparingthe curve: 02002-63 StudyNumber. FinalSolvent&TN-A#: Sera SPE validation MeOH/6308 [FinalVolumeofeach point:I 25l .... j: Curvepointnumber 02048-21-1 02048-21.2 C)2048-21-3 02048-21-4 ....... 02048-21-5 02048-21-6 02048-21-7 02048-21-8 02048-2t-9 02048-21-10 Amountofmix used(ul) 5 25 50 125 250 375 500 750 1000 1500 C6 0.100 0.500 1.00 "C7 0.102 I 0.511 I 1.02 2.50 5.00 7.50 10.00 15.00 "20.01 2.55 5.11 7.68 10.21 1513'7 20.43 30.0,1 I 30.64 C8 0.10:3' 0.517 1.03 "2.58 5.17 7.75 10.33 15,50 20.66 31.00 Analyte concentrationin every point (ppb) C9 0.101 0.807 1.01 C10 0.101 01505 t.01 Cll 0.103 0.515 ..... 1.03 C12 0,104 0_518' 1.04 2.54 5.07 7.61 10.14 15.22 20.29 2.53 5.05 7.58 10.10 15.15 20.20 2.58 5.15 7.73 10.31 15'.,_6 20.81 2.59 5.18 7.77 10.38 15.54 20.72 '30.43 I 30.30' ' 30.92 31;,08 THPFOS 0.102 0.512 1.02 2.56 5.12 7.68 10.24 15.37 20.49 30.73 THPFDS ' 0.100 0.501 1.00 2.51 5,01 7.52 10.02 15.03 20.04 I' 30.06 I PFOS 0.101 0.503 1.01 2.52 5.03 7.55 10.06 15,09 '20.12 30.18 01(/ co O _ Q ExacCt opyof Original _ /- Ildtlal Date ,,_ ...... "T" "': .... ..--r,.=..?............... ."=_-,.'.,'.-'T--_- ........ _" " " "- .... 3M ENVIRONMENTALLABORATORY E02-1039 ....i '_ I; "_ ', : i , : !ii : ; Standardmix trackingsheet Sere Super mixfor 8PE lOX MDV Prepdate: Analyst: 10/7/2002 OK Exp. 11/13102 0,_., /0' _'C_ Typeofthemix: sere supermix Targetanalytes: C6. C7. C8. C9. C10. C11.C12. THPFOS. THPFDS.PFOS Standardnumberforthe mix: 02002-63 StudyNumber:. FinalSolvent&TN-A#: jFtnalVolumeof the mix: Analyto C(] C7 'C8 C9 C10 C11 C12 THPFOS THPFDS PFOS concenlratloonf amountof concentratioonfthe Analytestandard enalyteIn standardstandardused analytaIn themix number mg/mJ(ppm} (ul) ng/ml(ppb) 02040-44 1042 12 500 02040-58 1344 9.5 511 02040-4,5 1230 10.5 517 02040-46 1266 10 507 02022-90 1010 12.5 505 0204065 1356 9.5 515 02040-55 1126 11.5 518 0204042 1348 9.5 512 02040-49 1002 12.5 501 02022-56 1048 12 503 Sere SPE validation MeOH/8308 I 25 ExactCopyof Origtnal Page 154 of 225 3M E_VIRONMENTALLABORATORY E02-1039 SINGLE COMPONENT PREPARATION LOG ' :_ Date: I0, I_0- 0 _J !! Analyst: 0 BookNo. 02 050 PageNo. 14 I DescflpUon: Ji:::}CT_p "_L#"_ :_ bL_,I"x _ _ 5? 'i 1 ='.',' StoCkNumber:.C)oQO- _3) _-- -_ _:= i,. ,....I. Weightor Volume Used: 2_ _ -i BalanceID: /_.)t_ Concentratioonr Purity: EO0,_. 0_' _ ,ziL. CorrectedWeight: _I/_ OtherCorrecffon Factors: tJ/% SolventandTN-A Number:. _eO -A " _ I_) ._ _" _ ._ ' I F,.., Volum.: FinalConcentration: _)Off_ ,f. d::! ExactCopyof 0rig[na/ ( _,lnc t_c/:t_lOL- Inltlal Data Reviewed /'Signat'_,"" _ 7/ jo Date( Page 155 of 225 3M ENVIRONMENTALLABORA'TORY .... E02-1039 v= _,L,:l 1 SINGLE COMPONENT PREPARATION LOG ,1 o,,e/o:.)c,_P.) BookN02o0. 50 ';4: ,;t.j , j :::i wo,,,,o,_o,u.3o7.0. %. -,(.4 ,.,...,o. f//t / <i ConcentrePtluornitoy:r -_tl f) ii;',!.,,:_ oo.-.w.,0/v.,A. _ij .,.,v._.. -_,-;_g ,.aJ + Storage Lo=ilon: _:_UOJ (_, :. ! ;.J L."JD Other CoFrarecctotirosn: //_/4 _,v..u.m,._,e.__.O. ///ql,' _ceJo I .,.._,.o....o, _ 7'-;xH_6- ExpimUon Date: tl.i;.o_J ExacCtopoyfOriginal L'i_c ICI',_)O-L.'- Inl_ g_ b Reviewed Sig_'natdia ,,,'-"_Z _ _'* ,_ / ioJ,/o Date Page 156 of 225 I II II__ _I ,, .... A II II I I llEl ...- 3M ENVIRONMENTAL LABORATORY E02-1039 "lq SINGLE COMPONENT PREPARATION LOG I Date: ]C ]C"C_ Book No. 02 050 Do,c,,,,o,- O.P ix i -:! Analyst: C_) Page No. 16 I -! i :/ Stock Number:. IO,gC,t'_. ,_ _ "_ I ; Weight or VoUlusmede: , I o_ "_" ] ,{,.,_ Balance ID: M/'_ " J ConcentraPtiounritoyr: , ,_ '= _"CC////_TrI_) I! Other CoFrarecctotirosn: /_///'31 _;_J "- ' ....: Fl.l_l,I V(il.IJITU_: _ _ i_t.l_ Storage Location: _I,_ I_ t_ }" Final ConP,:entltlltton: Expiration Date: Li'7)._.. C_" _ I"I //'/_" _'__) ! ExacCt opyofOriginal Initial Oa_ Reviewed by.,,#',vi"'--_'"S/X,,-'_'_ I_'_//_ ''_A Page 157 of 225 _._ __,_ i__. C _..... _ "" ": l l IBm. lif i l i m i m m m i R _ R l _ Description: Date Prepared: Prepared By: OO_Ue "E_'P,j ._-'2A6T_Co-r3M_L_O--_;-K -ic---C-_ _-/,'_ Io.:/.o2,jStorage Location: (J_Z,.._ Expiration Date: _Ot _ ,_(_ /I.r6,- C_/;_. MULTI-LEVEL PREPARATION LOG Book No. 02 048 PageNo. 21 Solvent: A[_-_--)/_ 3M Trace #: 7"A_-A-L_ _'_.-._,,_. Standard ID # 02048-21.1 02 048-21.2 02 048-21-3 02 048-214 02 048-21-5 02 046.21.6 02 048-21-7 02048-21-0 ''='_'____ Description ,., Description Compo ,_ntStandards ID # .. . Conc _/_O Expiration Date a. 1 Std 1 _. _" Std 2 ( _. 5" 5- 7. '_ ),_ Amount of Component Standard Added _,J_- Std 3 Std 4 Std 5 Std 6 Std 7 /:_Std 8 CV<, ........ L _ _=,V=,__on.,(_ _'_ _ ,_ ,...... -........-"."-.'-'> i -= FlnalConcentratlon.,,.-__@_..2... O. ( . O" _ | _."_ _ __ IC' J._"- Standard IO # 02049.2'1-9 02 04s-21-t0 02 048.21-1t 02 0411-21-1=02 048.21-1302046-21-14(}2N0.21-15 02 04s-21-1= _, ._ - ,,. ,-._, Descri.,p. tion, -,. _O _ C_ _..__---.- ----,-,,',------ _---'Tb,.,-,:-_'_-.: Component Standards - "Expiration Amount of Component Standard Added _ .... " Description ID # Conc Date Std 9 Std 10 Std 11 Std 12 Std 13 Std 14 Std 15 Std 16 __ C_ FinalVolume .j_S"- _"_ -"--'-----. _" bO to _ t:3 Reviewed / ._ / ....., 3M ENVIRONMENTAL LABORATORY E02- f 039 ,-_ .,.:I MULTI-COMPONENT PREPARATION LOG Book No. 02 002 Page No. 63 52"I . Oeac,p.o",_,:c"R'_.,2H/_'_ "-_'_A _'P _ so.,e.,t:/4_CH .! Date Prepared: _O- _'" C*,_ 3M Trace #: TN-A- (_ "_? i ' _ Prepared By: _")K_ storage,o=,.,:_,q_,j_. i Expiration Date: H" I :;_"C*-_ Standard.It:)# i iii 02 002-63 i i i m "... % Component=Standard '..l Corrected FinalConcentration ':-- Description ID # CPounrti,toyr ExDpiaratetion WVeoilguhmi'oer BalanceID Weight Tf,_.O | ii ,.- ,,,' :i C_.Ac_'b 4_ _.PTY .. C._ Ac;_ Cs ACi,_ _- r.r_ I 'tO _2c_- t_ _ _o_-, _.2.o3 12.,;kJ, -, _og- C_ /_c.;'_ 7__F-c,_5 iO.2CA-o- #u2._ _'.),q'_3 _1._/ :;5 it'-'-' . ,. 4-2. -_ ... ' %- r_.,_ ' _= T,t4?fib 5 _. Tf_ ... o _. .... ' _vv:, __- 'c4_i',,._,.o_I,z>_i >-o2_ -,i ,,,.1, L:Q,i.il _ c,_ -/_ _-c'2 Concentrationof the mix . ..,_ . - _,/ "Si0_nature. " _ "D_at' a/ Page 159 of 225 , : :_"_! i i "i "_ : i':'i'i '1 '_':' :! i , _::_i q'! 3M ENVIRONMENTAL LABORATQRY E02-1039 Test Control and Reference Substance Log I Substance trade name TDHA or reference # TCR Substance # TCR-99131-025 I Substance/chemical Trldecafluoroheptanolc Acid TCR # TCR-267 name: {Lot/batch #: PUI07219EU IRecelved from: Aldrich | I3M # NA I J IIExpiratJondate: 01/01/2005 I Amount received (wt. or ZS g, 37.1357 g gross wt. l VO ): {Initials: SRP [Date: 12/2111999 I I Number/size of 1130 mL plastic container [Shipper:. Courier ] containers: I [Condition: clear crystals IMSDS(y/n) Y'(.) N 1 IRetain 10'._3994_ {Date of Retain 92115/2000 I i.(_)'ArchivedlSfil_si_ance :N0t_Avaiiabie t , . ...... . . : :. -: 2 ..;J." . [Pudty: Records received: 99.5% LAG 4/24101 MSDS KJD 311100 Location of synthesis, fabdcation, or dedvation records: Std Location/Storage: Molecutar Formula: Comments NA F19, Frozen C6F13COOH Standard has been moved to Freezer 19 in room 347 KJD 06/07/00 Standard was stored at room temperature prior to 06107/00 LAC 12/22/00 Shipping comments: Corrosive, No 3M ID# - must ship ground small quantity sxcepUon (30 mug or less per vial). I.AS 10102/02 Attachment(s) I,_ L_ ExactCopyof Odglnal Data Page 160 of 225 _-; i _..,.. .t J J = L I i i i 1 .... !I?,_i i;_ I , : JAN 26 2808 89:48 FR ALDRZCH QC _':" 3M EtT_I_,_Sf_I980RA TC_,,,_4,,04 E02-I039 CERTIFICATEOF ANALYSIS _--,_,.=_ 3H ENVZRONHENTfd_OT'r P_T PQ NBR: PRODUCT NUH_ER: 34204.- LOT NUMBER: O'tRi?EU PRODUCT N_E_ TRt'_EC._FLUOROHEPT_NQIG ACZI), 99K FORMULA; G'rHFi30= FL_HULA WKZGHT: :364.06 _NFR_R1E;D8P_TRUH PLUORZNK Nf_ GAg LZQUZD _HROMATOGRalH-W _JALZTY P_J'INTROL A_CIEPTC_NE:ED_TE CONFORH_ TO b'FRUCTURK #=bid_L'AId]>_ A_ ILLUSTRATED tin PAGE 7?BB OF DZTZON ._, VOI.UHE t OF "TIE ALDRZGH LZBRARY OF FT-ZR _PEC11_". ONF_IRM.5TO 6TRUCTURE, 79._ X JUNE 1999 ExacCt opoyf0dgfnal AL1DRtCH CHKM_C_L L'J]CPNCf ,..I._UARY26 _ 2000 _ ch'umis_ helpingohamlscsin n_.searoh& Indusb3t /J-m_C_N_=r_tl_p_=_=_=_=_._._l b at: 4u__cl Axnm_a, i_,_mr mu__ _4 aldrichhemlr.ai o. --"_'"'_"-.--'-'--._.,.-_-,.--,, _ TOTAL PAGE. _4 _<_ Page 161 of 225 3M ENVIRONMENTALLABORATORY E02-1039 J ' 1 USELOG I "! p'ridecafluoroheptanoicAcid ; _IrDCHRA.99131-025 I 'i I Before withdrawn after or reasonforremoval) ID ; } withdrawal (massor vol) withdrawal 2 [GBraolsasnWcetIJDVo1l. BalAanmcnetI.D 2 BGarolasnscwetI/Dvo1l I Purpose(enterstandardnumber i Ba,l,_c, e I BalanceI lniUals ] Date [ 135.9s19g lO.OeSSg J35.a9_g IstandaOrdZO4O-Se p14 I" JRWW Joe/291:zoo2 I !_!i ii_ ExacCt opoyfOriginal Ill :_2 Page 162 of 225 ..,."... i":'.:.:_ r-r; :! _i:,,; !_j {_::_L,_. .o. 3M ENVIRONMENTAL LABORATORY E02-1 O39 _/.._l.> Date: u-'--_---" .2- s,.o,ecoiPo.,=p..T,:PAI_Tl.IOOQ, ._o.,o. o=04o Page No. 58 o.o._o.:7<,_,',/e/_,_x;. A..,..,1+,rJT-_-) Stock Number: p _'/..._J" ,,_'.J'" u.e.: Weightor Volume ,d-_'7. 0 Concentrationor Ba,,.,=o,:_ P'/"/ OtherCorrection correcWtee,lgh_. _, tP_ ?,Z2 SolventaNnduTNm-Aber.'/,",fM,,."-Till.--,#-z z _ V L_L Fin,Vi olume: ' _"D-" ./ FinaCl onc,nlrallon. v' /..]/./'Z_.p.p,. Storage Location: .2.,, ,Ar ,/_,/o _ #' _'#_ F_"._ ExplralionDate: _e//O 'i ExacCtopoyfOriginal _ '__ :i Jill ilj Sfgnaiure, Date Page 163 of 225 '.; 3M ENVIRONMENTAL LABORATORY E02-1039 SINGLE COMPONENT PREPARATION LOG .-' ..... Date: ,?'/_,6:,. i, q r . Book No. Page No. 02 040 58 o,,c,,7,:o,J::<,/:/...g.._./.;.,.#:.r.:.:J.,.,_.) 11_ .."... i= f:,_, !::i,:_:::i ..,.. Weight or Volume u,e= ,_- ObT.>'-? Concentration or Bale_,=D: _ / '-/ Other Correction Corrected Weight _-O _"_-- 4' / Solvent and TN-A Number:.'/_,,.:)/Z:]_ 7"/I/.7,__ _ ,]..__,/ .::J FinaVl olume: "_/3._ .)t F=alConcentration'/. 3 _=_./Je" -,] ._ Storage Location: .,2. ,_ .,_/o &_r' _'._,.. ,_/), ExplraUon Date: ,_t_,_ J_' ' ExacCt opyofOdglnal L Signature, .pil,o, Date " Page 163 of 225 t" i .....,. I: - i: 'i i i ii -_ i ii_ '_i'i::;:i , _":_ _ i_ '2 j _':: 3M ENVIRONMENTALLABORATORY E02-1039 "" 3M SPECIALTY MATERIALS MANUFACTURING DIVISION ANALYTICAL LABORATORY To: From: Lisa Stevenson - (8-5568) - ET&SS - 2-3E-09 Tom Kestner - (3-5633) - SMMD Analytical Lab - 236-2B-11 Request # GID:71638 Subject: Date: Characterization of TCR-99131-025 by 'H-NMR and '_-NMR Spectroscopy October 30, 2002 SAMPLE DESCRIPTION: TCR-99131-025, lot PU/07219EU from the Telomer project. Nominal )roduct = CF3(CFz).-CO2H, where average n = 5 (white solid). Sample Spectra#'s ExperimenDt escriptions TCR-99131-025 H71638.GID.401 400MHz ID 'H-NMRin a CFClj/acetone-ds_olventmixture+p-HFXcross integration/internasltd. TCR-99131-025 F71638.GID.401 376 MHzID IgF-NMRin a CFCl3/actone-dso_lventmixture+ p-HFXcross intqFation/intemasltd. ' TCR-9913!-025 F71638.GID.COSY.502 470MHz2D t_F-NMRCOSY(correlatedspectroscopye)xperimenint a CFCIv'acetonc-dso_lventmixture. OBJECTIVE: This sample was subjected to a combination of IH-NMR and 'gF-NMP,,spectral analyses to determine the purity of the nominal product. Special emphasis was also placed on attempting to identify and quantify any impurity components. EXPERIMENTAL: A portion of the sample was accurately weighed, spiked with a known amount of 1,4-bis(trifluoro- meixthtuylr)ebefnozresnuebse(pq-uHenFtX)a,naalnydsisthbeny NtoMtaIlLly dIin,;itsioallvoednei-ndiamCenFsCio1n3/adleu(1teDr)at4ed00acMetHonze tH(a-cNetMonRe-darn)d s3o7l6venMtHz )gF-NMR spectra were acquired at room temperature using a Varian UNITYplus 400 FT-NMR spectrometer. A two-dimensional (2D) WF-NMR correlated spectroscopy (COSY) experiment was also acquired using a Varian UNITY INOVA 500 FT-NMR spectrometer to facilitate assignment of tgF signals associated with various impurity components. This sample preparation method permitted the p-HFX to be used as either I) a IHftgF-_R. intemal stamlard to allow the calculation of the absolute weight percent concentrations of specific components, or 2) a 1H/19F-NM'Rcross integration standard to permit the cross correlation of the relative _H and 19Fsignal intensities for evaluation of the overall sample composition. RESULTS: - ".% The tH-NMR and tF-NMR spectral data indicated this sample was a relatively high purity form of the nominal product, CFa(CFz),-COzH, where the average value ofn = 5.05. Small amounts of a few impurity components, including probable isomers, were also assigned. A IH]IgF-NMR cross integration analysis technique was then used to calculate the relative weight percent concentrations of the identified components. The qualitative and quantitatiw," compositional results that were derived from the single trial IH/I'_F-NMR cross integration analysis are summarized below in TABLEd. The relative weight percent concentrations shown in TABLE-1 should be very close to their respective absolute weight percent values assuming no water was present in the sample. Trace amounts of numerous other unassigned components were also detected in the NMR spectra, but additional work would be needed in an effort to identify or quantify some of these other components. ExactCopyof Original Pagel of 2 _-- _ " Page 164 of 225 L-! r' I i ,i , , !: i ....,. iiii:i "_ . ; :i .i "'L" 3M ENVIRONMENTALLABORATORY E02-1039 October 30, 2002 31VSl MMD AnalyticalLab Request # GID:71638 .TCR-99131-025, Lot PU/07219EU: Telomer Project Copies of the NMR spectra are attached with the paper copy of this report for your reference. If you have any questions about these results, please let me know. Tom Kestner : MarkEllefson RickPayfcr RonPurcell WilliRacmagcn FilReefc_nIcseT:I63_.GID.TCR-L9a9Pt1U3I10-7O22159TEeUlom?erroject.DOC/lO| TABLE-1 Sample: TCR-99131-025, Lot PU/07219EU from the Telomer project. OverallCompositionRaelsultbsy II-I/tgF-NMRCrossIntegratiAonnalysis ComponentStructurtes IHflgF.NRM , Relative Weight% Concentrations (single trial analysis) "" CF_(CFz),-CO2H (where average n = 5.05) Probable internal branched isomers CF3(CFz)x-CF(CFa)-(CFz)y-COzH (where x_-0and assume x..+y= 3.05 for calculation purposes) Probable terminal isopropyl branched isomers (CF3)2-CF-(CF2)n-CO2H (assume n=3.05 for calculation purposes) A probable ether/acid as possible .. O-[CF2CF2-CO2H]2 .... 98.24% 0.87% 0.52% 0.13% " F F F' o 0.11% F.A--(--o/ . Probable FF C,H2,+2 saturated aliphatic hydroe_bons and functional aliphatic components, C,H2,+t-X, --0.11% where -X can be possible ether and ester functional groups. _ '.... Possible CF3(CF2),-COzCH3 0.021% ". (where average n = 5.05) 1. Traceamountsof numerous otherunassignedcomponentswerealsodetectedin the JgF-NMR spectrum. ExaCctopoyfoaginat _ Dal_ Page2of2 Page 165 of 225 E ' _i"i '"-_ i _,j i q ="_' i i_..i ."_ :! !i_i!_ .. , [_i!i] : 3M ENVIRONMENTALLABORATORY E02-1039 Certificate of Analysis Nominal Product: CF_(CF2),-CO2H,where average n _ 5 Tridecafluoroheptanoic acid Product Code: TCR.-99131-025, Lot PU/07219EU October 30, 2002 The sample of TCR-99131-025,. Lot PU/07219EU was analyzed using a combination of t9F-NIVIRand tH-NMR, spectral analysis techniques. The overall qualitative and quantitative compositional results that were derived from these combined analyses are summarized below in TABLE- 1. TABLE-1 S ample: TCR-99131-025, Lot PU/07219EU Overall Compositional Results by Combined 19F/IH-NMR Spectral Analyses ' Component Structures i UH/19F_NMR Relative Weight*/, Concentrations ....... ...... CF3(CFz).-CO2H (sing!_etrial analysis) <98.2% (where average n = 5.05) "' Probable internal branched isomers Purity 0.87% CF3(CF2)x-CF(CF3)-(CF2)),-CO2H (where x_-Oand assume x+/= 3.05 for calculation purl_oses) Probable terminal isopropyl branched isomers .... 0.52% (CF3h-CF-(CF2).-CO2H (assume n=3.05 for calculation purposes) .. A probable 'e'ther/aeidas possible O-[CF2CF2-CO2H]2 0.13% F v F .O. ' 0.11% F_o_ Probable FF c.H2.+2 saturated aliphatie hydrocarbons and _0.11% _---.. functional aliphatic components, C.H_.+I-X, where -X can be possible ether and ester functional .groups. Possible CF3(CF2),-CO2CH3 0.021% (where average n = 5.05) 1. Traceamountsof numerousotherunassignedcomponentswerealsodetectedm'theNMRspectra. "= Tom Kestner ExactCOpyof Original r Page 1 of I FileReference:Cola TCR-99131-025Lot PU-07219EU.doc Page 166 of 225 :1. ' . ,i/ :TI_i 1 ' .... '", i i,_,i} .. 3M E-ZNWRONMENTAL LABORATORY E02-1039 Test Control and Reference Substance Log Substance trade name _r reference # Substance/T:hemical _ame: Pentadecafluorooctanoic acid Pentadecafluorooctanoic acid .ot/batch #: :3M # 1210002 I Expiration date: Initials: Number/size of containers: J I IOK Jl-t0ml I amber glass vial Condition: Retain _white crystals . t TCR Substance # ..... TCR # TCR-617 TCR-817 Received from: Oakwood Products Amount received (wt. or 5g _ol): Date: Shipper:. 0711912002 Courier VISDS (y/n) Date of Retain t Y t. ) N ! (,_ :Aibhived/Substance. Not:/(_ailable"1 ... ?_.; :.. . ,, :.... ...'--, . .,.-_:.'__:_;'/:.- Purity: 99.51%, updated 10130102OK, NMR analysis Records received: MSDS I'ocation of synthesis,' fabdcation, or derivation records: Certificate of Analysis I_AS 07/25102 Oakwood products Std Location/Storage: Molecular Formula: Comments Room Tamp, TCR-C01 C8HF1502 Corrosive Solid LAS 07119/02 Shipping comments: No 3M ID# - must ship ground small quantity exception or less per vial). LAS 10102/02 Attachment(s) L_ TCR-617.pdf , .. (30 mL/g ExactCopyof Original "" Cwlc _olal! Iilllil L !i Page 167 of 225 , =; i an ] III I II J , # . .. ', ,,_ ' : 'J .... i i , ...:.. } .... .' .., -'. v ' ,. ",, .'. ' I ... .. '.... ._: _ . ", _', ,, I . : _' :" : "" ,.,,,,. !?: ..,,: ':':' :"" .... !:,: . ""_'" : .." ":".. ' o ...,. :i: '"",' "' !:'.... Oakvcood Products, Inc. I741 Old Dtmbax l'Load West Colambia, SC 29172 ]rhone I1_03) 739-8800 Fax(803)739-6957 ." CERTIFICATE OF ANALYSIS Date: XS-ltflo02 Material: Pentadecaflum'o oc tanoic add .. Cat.No.: 1319 Cas No.: [335-67-:1] ]Lot No.: 210002 Assay: 97+% by NaOH tilxation .;:'.-.. .,.' ,' ... " ' ....".. "v" ..:'r: . -': ": r. ': :....:.. ,. .( 4; _,. __;'; :f ." : "" .,._ . ' .-.... ;,, ,o ., ..,.. "' :.- ,..-, . :,...'....... o,.. '.X , "-';:., ""7"_. i.''.":'' .., ....., . :, :: - .: ..-,, ,., ': "': j . "_ . . ... .._. ::i:_i if : ..,I +" ::.. ,. :. " _ ' .,. .'. , "," ;, . i ;" : ,.:l...:...":.]..'.'. ; . _ ,. . i:, . ]. ,.'= Appeaxa_e: x_qi_te solid Melting Point:. 59-61_'C .: _:..-.. ; ".,'!' __ :i !::i...:,.,........ :: i _' umet Yah ',--"QC M_a e,'=-_r :..""'.. :. ;.. :: ! :._: ., . "': ..... . } ' ":""?,'o." '*" _acCt opyofOriginal ._ :., at.'. ":. , " _ Date :.i .Y, ;t ._.',.. ,3 . .... ;| .='._.'.. "-.', ., :_ ..:..:. _. : ":" ' "" "'..:: " :"."'- ; .... .,. , :..", , .] ._,_. , .. .A.- Page 168 of 225 3M ENVIRONMENTAL LABORATORY E02-1039 I .... entadecafluorooctanoi acid , JPentadecafluoroocta nolo acid I _CR'617 USE LOG _-_ J Before withdrawn after or reason for removal) ID ID I withdrawal (mass or vol) withdrawal 1 2 IGBroalsasncWet.INDol1. BalaAnmcent.ID 2 BGarlaonscsewtI/Dvo1l I Purpose (enter standard number I Balance I Balance I Initials I Date Ils.._asg Io.oslrg Ils.48eee Istanda0rd2040-45 t914, l" IRww Ioa/ls/2002 :/i J r ,'!::j ExacCt opyof 0dglna] ,i 1 Page 169 of 225 :, ! oa,,: 7,_-"_._ :i ,_v,/ Analyst: : } * 3M ENVIRONMENTALLABORATORY E02-1039 SINGLE COMPONENT PREPARATION LOG BooNko.020,0 Page No. 45 . =,, Stock Number:. TC_- _"// :i ,I ....i .- Weight or Volume ;'"_ U,ed.: _. Ol/-._.2 Concentration or Pudty: _. _ !! Corrected Weight: _),' D_//.S"_ / B_l__nDo:o. 7 : '7/' Other Correction Factors: /4-// Solvent and TN-A Number: ,_C_, ' _/.'_/7_,_,--/'_ -/__.2--.'i2 I',:,.: Final Volume: 5-o I _ ,.., Final Concentration: L2.._":,_,.,, : StorageLocatlon: 9_2 :/f:.._,[-"-.//2o.,._.,._.,'I i i L'_._ Expiration Date: _?S'S'_._ ExacCt opyofOriginal 8kjnaturo, Dato " Page 170 of 225 3M ENVIRONMENTALLABORATORY E02-1039 3M SPECIALTY MATERIALS MANUFACTURING DIVISION ANALYTICAL LABORATORY _::i t To___2 Lisa Stevenson - (8-5568) - ET&SS - 2-3E-09 Request # GID:71638 From: Tom Kestner - (3-5633) - SMMD Analytical Lab - 236-2B-11 Subject: Date: Characterization of TCR-617 by IH-NMR, 19F-NMR, and LC/MS Analyses October 28, 2002: Updated R,:port - LC/MS Analysis for Impurities :if_ ! ...._ I i_!:_];-iJ ' ii_ I!':!_ '_ _ i :_ i .?! SAMPLE DESCRIPTION: TCR-617, lot 210002 from .theTelomer project. Nominal product = CF3(CF2)r-CO2H, where average n _ 6 (white powder). Sample Spectra#'s ExperimenDt.escriptions TCR-617 TCR-617 UPDATE: HT1638.GID.501 500 MHz'H-NMRinaeetone-dsnolvent+p-HFXcrossintesration/internasltd. F71638.GID.501 470MHz t_F-NMRinaeetone-d_solvent+ p-HFXcrossintegration/internasltd. ' After the initial NMR compositional analysis report was issued to you on 10-24-02, Joel Miller performed a ! qualitative LC/MS analysis to assist in the assignment of some of the impurity components in the sample of i TCR-617. This updated reported summarizes the new information regarding the identifies and :_t concentrations for some of the impurity components. You will notice that the tentatively assigned olefin : structure from the original report is now replaced with a probable (C6FI30)-CO2H ether acid. r OBJECTIVE: This sample was subjected to a combination of _H-NMR and 19F-NMR spectral analyses to determine the purity of the nominal product. Special emphasis was also placed on attempting to identify and quantify any i iinmpthueritayssicgonmmpeonntenotsf. impTuhreitysamcopmlepownaesntasl.so JgoievlenasktoedJotehlatMIiilnlecrorfpoorraatqeuatlhietaqtiuvaelitLaCti/vMe Sinafonramlyastiisonto farossmist his LC/MS analysis in this updated report. "' FT-NMR EXPERIMENTAL: 'i:t .;[ : _ ....". ; .I "; A portion of the sample was accurately weighed, spiked with a known amount of 1,4-bis(trifluoromethyl)benzene (p-HFX), and then totally di.,molvedin deuterated acetone (acetone-d6) for subsequent analysis by NMR. A 500 MHz _H-NMR spectrum and a 470 MHz 'gF-NlVlRspectrum were acquired at room temperature using a Varian UNITY INOVA 500 FT-NMR spectrometer. This sample prel_aration method permitted the p-I-I'FXto be used as either 1) a IH/IF-NMR internal standard to allow the calculation of the absolute weight percent concentrations of specific components, or 2) a IH/19F-NMR cross integration standard to permit the cross correlation of the relative tH and 19Fsignal intensities for evaluation of the overall sample composition. RESULTS: The combined )H-NMR, 19F-NMR, and LC/MS analyses indicated this sample was a high purity form of the nominal product, CF_(CF2),-COzH, where the average value ofn = 6.02. Small amounts era few impurity components, including probable isomers, were also assigned. A tI-UmF-N'MRcross integration analysis technique was then used to calculate the relative weight percent concentrations of the identified components. The qualitative and quantitative compositional results that were derived from the combined single trial IH/J'_F-NMR cross integration analysis, and the qualitative LC/MS analysis, are summarized below in TABLE-1. The relative weight percent concentrations shown in TABLE-I should be very close to their respective absolute weight percent values assuming no water was present in the sample. PageI of 2 [xa_ Copyof Orfginal ,o_au.o Initial n,_. Page 171 of 225 i ,i !_ I i !I -: i :] '.'l ,t !_!!] !_''!,,._',_ _1 'i '.i 3M ENVIRONMENTALLABORATORY E02-1039 :'i , _ ,:i l :! i : _i ! I October 28, 2002 3M SMMD Analytical LabRequest # GID:71638 Updated Report - LC/MS Analysis for Impurities in TCRgil7, Lot 210002 RESULTS (cont.): Trace amounts of a few other unassigned components were also detected in the NMR spectra, but additional work would be needed in an effort to identify or quantify these other components. Copies of the NMR spectra are attached with the paper copy of this report for your reference. If you have any questions about these updated results, please let me know. Tom Kestner c: JRoicekMlPiallyefrcr RonPurcell WilliamReag=n FileReferencIes;71638.OlO.UlgRla,.t,eudlts_TCR-61271..0.L0o0t2TelomePtrojeeLDO0CI /I TABLE-1 Sample: TCR-617, Lot 210002 from the Telomer project. Overall Compositional Results by IH/19F-NMR. Component Structures I CROSISnte_ation and LC/MS Analyses NMR Relative Weight% Concentrations ...... (single trial measurement) CF3(CF2)n-CO2H where average n = 6.02 by lgF-NMR. LC/MS showed n=6 (major), n=5, n--4 (minors). Probable (CFa)2-CF-(CF2)n-CO2H 99.52% 0.39% assume n----4for calculation pro'po, ses Probable (C6FI30)-CO2H aeyelic ether acid as possible CF3CF2-O-CF(CF3)-CFzCF2-CO2H Possible CF3(CFz)x-CF(CF3)-(CF2)y-CO2H where x;_0,y_0 and assume x+y : 4 for calculation purposes Possible (CF3)3-C-(CF2).-CO.,H assume n=3 for calculation purposes PossibleC.H:.,2 saturatedaliphatichydrocarbons 1. Traceamountsof otherunassignedcomponeaatwserealsodetectedin theNMR.spectra, 0.057% 0.019% 0.013% 0.0079% \ -"%. ,i Page2 of 2 ExactCopyof Original tel :_ll(,ft..- _ _a_i-- Page 172 of 225 ..=.. 3M ENVIRONMENTAL LABORA TORY : E02-1039 i ' :. _ i f .,i ' Ii ' !_ I ! :;Ii] ;:"_ !_:it: _,, ': !' ; . _], : ,i ; :! ; , iI L Certificate of Analysis Nominal Product: CFj(CF2)n-CO2H, where average n _ 6 Pentadecafluorooctanoic acid Product Code: TCR-617, Lot 210002 October 28, 2002 Tom Kesmer and Joel Miller The sample of TCR-617, lot 210002 was analyzed using a combination of 19F-lqM_, IH-NMR, and LC/MS analysis techniques. The overall qualitative and quantitative compositional results that were derivedfromthesecombinedanalyseasre summarizedbelowinTABLE-I. TABLE-1 Quantitative Sample: and Qualitative Compositional Component Structures t TCR-617,Lot210002 Results by Combined 19F/IH-NMR and LC/MS Analyses IH/19F__vl_q. Relative Weight% Concentrations (single triaalnalysis) CF3(CF2),-CO2H where average n = 6.02 by IOF-NMR. LC/blS showed n=6 (major), n=5, n=4 (minors). Probable (CF3)z-CF-(CF2),-CO2H assume n=4 for calculation purposes Probable(C6F,30)-CO2Hacyclic etheracid as possiblCeF3CF2-O-CF(CF3)-CF2CF2-CO2H PossiblCeF3(CFz)x-CF(CF3)-(CF2)r-CO2H where x;_0,yea) and assume x+7 = 4 for calculation purposes Possible (CF3)3-C-(CF2)a-CO2H <99.51% Purity 0.39% 0.057% 0.019% 0.013% a_surne n=3 or calculation purposes Possible C.H2.+2 saturated aliphatio hydrocarbons 1. Trace amounts of other unassigned components were also detected in the NMR spectra. 0.0079% #, -- Page I of I File Reference: CofA_TCR-617_Lot 210002.doc Exa_Copoyf Orfglna) i_ Oato Page 173 of 225 i '_ '/:':I ! ..... ; ; :_ , ' ;i 'i i..J i! ' r_c .?:.., 3M ENVIRONMENTALLABORATORY E02-1039 Test Control and Reference Substance Log !Substancetrade name Heptadecafluorononanoicacid TCR Substanc#e TCR-618 or reference# Substance/chemical name: Heptadecafluorononanoicacid TCR # TCR-618 ]Lot/batch#: 3M_ Expirationdate: Initials: H7588 NA OK IReceivedfrom: I Amountreceived(wL or Ivo): IDate: OakwoodProducts 5g 0711912002 Number/sizeof containers: 1-50ml plasticjar IShipper:. I Courier Condition: _hite crystals. JMSDS(y/n) Y (.) N I Retain JDateof Retain I i(_i,Aichived/Sub'starice_oi:-_vai,i-_.i_ :Purity: 198.02% updated1013010O2 K, NMRanalysis :Recordsreceived: MSD$ Locationof synthesis, fabrication,or derivation records: Certiflcatof Analysis LAS07/25102 Oakwood Products Std Location/Storage: ( RoomTemp, TCR-C01 MolecularFormula: Comments IC9HFt702 :Shippingcomments: Corrosivesolid, No3M ID# - must ship groundsmall quantity exception(30 mug or lessper vial). LAS 10102/02 Attachment(s) _._ TCR-618.pdf __ ExacCt opoyfOr/g_nal ':: ] i L Page 174 of 225 ,_;, . ,_-,w ),?_d,?>_-..... ' :_ ' Oakwood Products, 1741 Oht Dunbar Road _'VesCt t_[uu,.biaS, C29172 ['hone {8t_3)"Dg-SSO0 Fax (803) 730-6957 Ir_c. CERTIFICATE OF ANALYSIS :: ""' Date: 27-Nov-01 ' , ! .: ' .: _ " ' .::., .i : .... '.i ".:' , .., r._ . ' :L,,. ',i:i _. " i: 4, Ma{ex4nI. Heptadeciffluorortona_mic add Cat.No.: 2263 CaS No,: Lot No.: [375-95-2] H7568 Assay: 99*% by NaOH titration .,,, ,,._..-, : " ' Appearance: Off-white _olid _. :,::'.:":., Melting Polxtt: 57-62C i" L ,. ' '..: i.'. ,, ,.. : _. ,. : ::'. ., ::' 4 ::::",' ;' ':...... ,_,. 4. " " :,.? , ' '3 , _, . ' ' :..... . ... , ., : " , . ., :" ":,,i." '" ". X k :' _: '" ? ? :..: :', ExacCt op-oy:l:O:':r,i'_inal;'" c>c _ ._ ',', '.::: :" ,_ ., , , '.' ,._ ':, , "' ,. Page 175 ot/'225 ; i,, I eptadecafJuorononanoic acid :i _eptadecafluorononanoic acid 'i IrcR-818 3M ENVIRONMENTALALBORATORY E02-1039 USE LOG o,.o,,w,.,__on,,.. withdrawal Balance ID 1 (mass or vol) Balance ID 2 BOwaliathn,dc'reaow|Da,l1',I,,,",,,.v,ro,,,=o,on'e,'sI="",a,,,',_c,o,',,""a,,,a"nor_,',:n"",,=1,, 20.8649g 0.0677g 120.7972g Istandard02o4o.4s _14 l" IRWW Io511_00Z ;! ,7,1 _!(i ExacCt opyof Original _! _ oat= t :J Page 176 of 225 i,." 3M ENVIRONMENTALLABORATORY i E02-1039 7 SINGLE COMPONENT PREPARATION LOG I Oata:Y/_- ! _ookNoo2.o,o ' Page No. 46 Analyst: #vIJ i i Description:..I1;,,7_ olaf,//,<o..,,,/__.,,.,,;,2..i/.o,<.7,7,.(>"i,si'_,l_-,c,_o,# :'", L,t [ ; Weight or Volume Concentration or Purity: #/_ .5 Corrected Weight: #/_,, O_dc/# h:G FinaVlolume: ! ,_.)'--#;"t Other Correction Factors" A//_" Solvent and TN-A Number:. tlL'/l,'_.,'.,//_-'ll/,-#.l#,__._,2.._._.., . FinalConcentration:L .2 6 ,_ ,,F,# ExacCt opyofOriginal .... Initial Date Reviewedby:,..._._.%A, c' Signature, il_/r)_. Oat_ Page 177of 225 ,! :_"iill :-_ i ':/_i i :J -_ !__.t !'[ ; ! J _i_ ::_" ..'. -? i) .i _ ii 3M ENVIRONMENTALLABORATORY E02-1039 3M SPECIALTY MATERIALS MANUFACTURING DIVISION ANALYTICAL LABORATORY To: From: Lisa Stevenson-(8-5568) - ET&SS- 2-3E-09 Tom Kestner - (3-5633) - SMMD Analytical Lab - 236-2B-l 1 Request # GID:71638 Subiect: Date: Characterization of TCR-618 by IH-NMR and 19F-NMRSpectroscopy October 28, 2002 SAMPLE DESCRIPTION: TCR-618, lot H7568 fromthe Telomerproject. Nominal.product = CF3(CF):,-CO2H,where average n _ 7 (white solid). , Sample Spectr#a's ExperimenDtescriptions TCR-618 H71638.GID.402 400MHzIH-I_MiRnacetone-dso_lven+t p-HFXcrossintegration/intexsntd_. TCR-618 F71638.GID.402/.403376M_Izv_F-NMiRnacetone-ds_olven+t p-HFXcrossintegration/intersntda.l OBJECTIVE: t This sample was subjected to a combinationof IH-NMR and IgF-NMRspectral analysesto determine the purity of the nominal product. Special emph:tsiswas also placedon attemptingto identify and quantify any impurity components. EXPERIMENTAL: A portion of the sample was accuratelyweighed, spikedwith a known amountof 1,4-bis(trifiuoromethyl)benzene (p-HFX), and then totallydissolved in deuteratedacetone (acetone-de)for subsequent analysis by NMR. A 400 MHz _H-NMRspectrum and 376 MHz tgF-NMRspectra were acquired at room temperature using a Varian UNTI'Yplus400 FT-NMR spectrometer. This sample preparation method permitted the p-HFX to be used as either 1) a IH/=gF-NMRinternal standard to allowthe calculation of the absolute weight percent concentrationsof specific components,or 2) a 1H/tgF-NMRcross integration standard to permit the cross correlationof therelative IHand 19Fsignal intensitiesfor evaluationof the overall sample composition. RESULTS: Thv IH-bTM17,.and 19F-N'DtI_ spectral data indicated this sample was a high purity form ofthe nominal product, CF3(CF2)a-CO2I-wI,here the average value ofn = 6.953. Small amounts of a few impurity components were also assigned. A II-I/I_F-NMRcross integration analysistechnique was then used to calculate the relative weight percentconcentrationsof the identifiedcomponents. The qualitative and quantitative compositional results that were derived from the single trial H/19F-NMRcross integration analysis are summarized below in TABLE-1. The relative weight percent concentrations shown in -". TAB LE-1should be very close to their respective absolute weight percent values assuming no water was present in the sanaple. Trace amounts of a few other unassigned componentswere also detected in the NMR spectra, but additional work would be needed in an effort to identifyor quantify these other components. Copies of the NMR spectra are attachedwith the paper copy of this report for your reference. If you have any questions about these results, please let me know. ExacCt opyofOriginal. Page 1 of 2 _ l_ats Page 178 of 225 !?! I ii<i ...q. i ' __ : ',,,,)_,_ .... '::_I _:_) :] : )_i_:ii _ i_ii"i L,:J JL _-.,_ October 28, 2002 3M ENVIRONMENTALLABORATORY E02-1039 ".. 3M SMMD Analytical Lab Request # GID:71638 TCR-618, Lot I-I7568:Telorner Project Tom Kestner e: RickPayfer RonPurcell WilliamReagen Fil,R":ferencIes:TI6311.GID.TCR-1611785_6LT8oetlomPerrojectDOC./100 TABLE-1 Sample: TCR-618, Lot H7568 from the Telomer project. Overall Compositional Results by tI-I/19F-NMRCross Intesration Analysis Component Structures t NMR Relative Weight% Concentrations (single trial measurement) CF3(CF2),-CO2H where averase n = 6.955 Probable H-(CF2)a-CO2H assume n = 8 for calculation purposes Probable CF3(CF2)n-CO2CH3 where average n = 6.955 A probable cyclic ether F O "C'F4 98.02% 1.10% 0.65% 0.20% F F as possible F t_ or similar Total inorganic fluoride (at least 3-t_pes), including some possible HF Possible C,H2n+2saturated aliphatie hydrocarbons 0.021% 0.014% 1. Traceamountsof othermmssignedcomponentws er,e alsodetectedintheNMRspectra. Page 2 of2 ExactCopyof 0 rigihal -._)_C_ lo/.L._.O-L." blit[al Date Page 179 of 225 3M ENVIRONMENTALLABORATORY E02-1039 :i i i ._i , _ ! !_ ': i i_ ,:! .... J Certificate of Analysis Nominal Product: CFa(CF2)a-CO2H,where average n _ 7 Heptadecafluorononaaoic acid Product Code: TCR-618, Lot H7568 October 28, 2002 Tom Kestner The sample of TCR-618, lot H7568 was analyzed using a combination of 19F-NMR and IH-NMR spectral analysis techniques. The overall qualitative and quantitative compositional results that were derived from these combined analyses ate summarized below in TABLE-1. TABLE-1 , Sample: TCR-618, Lot H756g Quantitative Compositional Results by Combined 19F/tH-NMR Spectral Analyses Component Structures ' 'H/19F.NM R Relative Weight% Concentrations (sinl_le trial anal)sis) CFa(CF2),-CO2H where average n = 6.955 Probable H-(CF2),-CO2H assume n = 8 for calculation purposes Probable CFa(CF2)n-CO2CH3 where average n = 6.955 A probable cyclic ether F0 F F "C4F9 F F <98.02% Purity 1.10% 0.65% 0.20% as possible F F orsimilar Total inorganic fluoride (at least 3-type:Q, 0.021% ineludin_ some possible H-F Probable C, H2,+2 saturated aliphatic hydrocarbons 0.014% ,. 1. Trace amountsof otherunassignedcomponentws erealsodetectedintheNMRspeclra. ExactCopyof Original Pagel of I FileReference:CofA_TCR-618_LoHt7568.doe Page 180 of 225 i! '- i '. _,.; .i i ; _ "3 i. i i___! I 3M ENVIRONMENTAL LABORATORY E02-1039 Test Controland ReferenceSubstanceLog Substance trade name or reference # Substance/chemical name: _lonadecafluorodecanolc acid Nonadecafluorodacanolc acid TCR Substance # TCR # SD036 TCR-36 Lot/batch #: 3M # IR11 K ]10#2284 jRecelved from: I IOakwood Products Inc 4/2/99 I I I Expiration date: niUals: Number/size of containers: 12/0112010 IJCP 111120mL plastic bottle i voAmunrte)c:eived (wt.or 25g,43.3264 g gross wt. I jDate: 104/27/1999 I I JShipper: N/A , I Condition: Jwhlte solid MCH 07122J99 JMSDS (y/n) Jg Y _..) N J Retain j0.1874g [Date of Retain 11011811999 I "(:')/_rchived)_ubstance Noti:Avaif_,_le_j Purity: 198.01%,updated t0/30102 OK NMR analysis I Records received: IMSDS,Certificate of Analysis I oc,ooolu,.,on.o o.s I fabrication, or derivation ecords: iStd Location/Storage: JF19, Frozen I Molecular Formula: I C9F19COOH I Comments TN-A-2451 prior to 4/27199 JCP 04/27199 Standard ha,= been moved to Freezer 19 In room 347 KJD 06/06/00 Standard was stored at room temperature prior to 06/06100. LAC 12/19/00 Attachment(s) _ sd036msds.pdsf d036cofa.pdf EXaCt Copy of Original . Iiddai Oats Page 181 of 225 .... = _,_ i!,,<! i '..,._.j.. ,, ,_ i ;-,_ .! _i_ '_ _t ,_::i _!i!:?'i 3M ENVIRONMENTALLABORATORY E02-1039 3M SPECIALTY MATERIALS MANUFACTURING DIVISION ANALYTICAL LABORATORY To: From: Lisa Stevenson - (8-5568) - ET&SS - 2-3E-09 Tom Kestner - (3-5633) - SMMD Analytical Lab - 236-2B- 1! Request # GID:71638 Subiect: Date: Characterization of SD036 by tH-NMR and tgF-NMR Spectroscopy October 28, 2002 SAMPLE DESCRIPTION: SD036, lot R1 IK from the Telomer project. Nominal product = CF3(CF2),-CO2H, where average n _ 8 (white powder). Sample Spectra #'$ ExperimentDescriptiom SD036 SD036 H71638.GID.403 400 MHztH-NMRin aeetone-d_solvent+ p-HFXcrossintegration/imernn/std. F71638.GID.404 376 MHz "F-NMRinacetone-d,solvent+p-HFXcrossintegration/internasltd. OBJECTIVE: This sample was subjected to a combination of tH-NMR and _gF-NMR spectral analyses to determine the purity of the nominal product. Special emphasis was also placed on attempting to identify and quantify any impurity components. EXPERIMENTAL: A portion of the sample was accurately weighed, spiked with a known amount of 1,4-bis(tfifluoro- ' methyl)benzene (p-HFX), and then totally dissolved in deuterated acetone (acetone-d6) for subsequent analysis by NMR. A 400 MHz JH-NMR spectrum and a 376 MHz 19F-NMR spectrum were acquired at room temperature using a Varian UNITYplu_ 400 FT-NMR spectrometer. This sample preparation method permitted the p-HFX to be used as either 1) a IH/_gF-NMR internal standard to allow the calculation of the absolute weight percent concentrations of specific components, or 2) a IH/tgF-NMR cross integration standard to permit the cross correlation of the relative tH and 19Fsignal intensities for evaluation of the overall sample composition. RESULTS: The II"I-F_,fRand 19F-N'M'Rspectral data indicated thls sample was a relatively high purity form of the nominal product, CF3(CF2)n-CO2H, where the average value ofn = 7.85. Small amounts of a few impurity components, including probable isomers, were also assigned. A tH/tgF-Nl_fl_,cross integration analysis technique was then used to calculate the relative weight percent concentrations of the identified components. The qualitative and quantitative compositional results that were derived from the single trial , IH/19F-NMR cross integration analysis are summarized below in TABLE-1. The relative weight percent -'" concentrations shown in TABLE-1 should be very close to their respective absolute weight percent values assuming no water was present in the sample. Trace amounts era numerous other unassigned components were also detected in the IF-NMR spectrum, but additional work would be needed in an effort to identify or quantify some of these other components. Copies of the NMR spectra are attached with the paper copy of this report tbr your reference. any questions about these results, please let me know. If you have ExactCopyof Original Page l of 2 Page 182 of 225 '! i_!*i "_?i i _i ' _,:,i 'J !i --j .:_J _ I'::_!_I i_!ill ]"_ 3M ENVIRONMENTAL LABORATORY E02-1039 October 28, 2002 3M SMMD Analytical Lab Request # GID:71638 SD036,LotRI 1K:TelomerProject Tom Kestner c: RRoicnkPPuarycfeelrl William Rcagen I=il Reference: Is71638.GID.SD036_Lot RI I K..Tclomer PmjccLI:X3C/I OI TABLE-I Sample: SD036, Lot R11K from the Telomerproject. Overall CompositionalR.esu!tsby 1I-I/19F-NMCRr.oss Integration Analysis Component Structures t NMR Relative Weight% Concentrations (single trial measurement) CF3(CF2).-CO2H where avera[[en = 7.85 98.01% , Probable CF3(CF2)x-CF(CF3)-(CF2)y-CO2H wherex_0 andassumex+y = 5.85for calculationpurposes Probable (CF3)z-CF-(CF2)n-CO.,H assume n=5.85 for calculationpurposes Probable 1.41% 0.36% 0.12% F F F I assume n=3.85 Pfororbcaablcleulationpo urposes F II F FF 0.060% FF B F CF OH assume n=2.85 for calculationpurposes Probable CnI-Iln+2 saturated aliphatie b7dmcaxbons Total inorganicfluoride 0,026% _0.0095% 1. Trace amounts of numerous otherunassigned components were also detected in the J'JF-NMRspectnm_ ,' ExacCt opyof Original J Page 2 of 2 Page 183 of 225 Ot::_KWIP3R0D013UCTS Fax:803-?39-6957 Feb3_ EZ_(_oN,_'m- LAe_"_",,R,_Y -I :,.] ...'. i.' I ,., _ ,!?' _, ,_i 17_ Old _b_ bad WestColumbia,$C 2g_l._. i,i,.ot_(sos) (_ Oa3_ood l'roduets,Inc. CERTIFICATE OF ANALYSIS Date: 11-Feb-00 Material: Nonadec_luorodecanoi add Cas No.: [335-76-2] Lot No.: RI::I..K Ashy: Products mare volatile than I'etfluorodecanoi,' aJcd <1% Per1uomdet_onlc acid 98/orain. Products less volatile _an Perfluo_decanoic acid <1% Appea_anc_ White solid Melting Point= 83-85"C EXacCt opyofOdglnal _ i_Lm.. Page 185 of 225 3M E'hlVlRONMENTALLABORATORY E02-1039 I USE LOG ] " "; _lonadecafluorodecanoicacid ,', ' __SoDna0d36acafluorodacanoicacid I Before withdrawn after orreasonforremoval) IO withdrawal (massor vol) withdrawal 2 Balance D 1 BalanceID 2 BalanceID 1 J33.9944g ]0;:i'19Sg J33.8784g ]standard02022.90 J914 J. [RWW [07102/2002 I t , ExacCt opyof 0dginal N Page 186 of 225 '",_I _,=,:, i..i 3M ENVIRONMENTALLABORA TORY E02-1039 Date: ,Z.. ,_,,=ys_. ,/,_j _ SINGLECOMPONENTPREPARATIONLOG De..,,,o,.P F/J._ _ /_, /L BookNo. 02 022 F',_eNo. 90 sto=.u.,be_. ,.5/) d3_ Weight orVolume Used:,. f'_/,_ Concentrationor BalanceID: _/_-/ Other Corre_on _!t ..... Co.,,_e,_W/e__Ig.,_D_'I Solventand TN-A Number./_I,-,_/,Zx.,,z.-..,_#-,,_z. ." Final Volume: . /_. / ,,o,._,,o. =,o,z.:.jF/_.,, ! :_,_.i ...:.. Final Concentration: /Z_/_ ," .tt'_ _,,,,,o,o,,,.,. /_/D,_ ExacCt opyofOriginal InJr/el Date U S_nature, /1 Date Page 187 of 225 !i:'I ":;'i ,i ':',i :: _. _:._.,,i "__'_ : I , ..- ._ ?--! 3M ENVIRONMENTAL LABORA TORY E02-1039 Test Controland ReferenceSubstanceLog Substance trade name =erfluoroundecanoic acid TCR Substance # TCR-619 or reference # Substance/chemical name: Perfluoroundacanoic acid TCR # TCR-619 Lot/batch #:. 3M # JU11N JNA Received from: Oakwood products I I Expiration date: _1 vol):Amruencteived (wt. or 5 g I Initials: IOK Dale: 107119/2002 I I Ncounmtabienre/srisz:e of 1-10ml amber glass vial Shipper:. IjCourier I Condition: Iwhits crystals. MSDS (y/n) I(ID Y (..) N '1 Retain I Date of Retain I I ! .("_ArchivedlSubstanc:e,'i_0At;vaiiab'lei:J Purity: Records received: Location of synthesis, fabrication, or derivation records: Std Location/Storage: Molecular Formula: Comments I >99% LAS 07125/02 MSDS Certificate of Analysis Oakwood products I,AS 07/25102 I Room Tamp, TCR-C01 I C11HF2102 Shipping comments: Harmful solid, No 3M ID# - must ship ground '(30 mug or less per vial). LAS t0/02/02 small quantity exception Attachment(s) L_ TCR-619.pdf Exact Copy of Orlgl_ Page 188 of 225 3M ENVIRONMENTALLABORATORY E02-1039 j "., 3M SPECIALTY MATERIALS MANUFACTURING DIVISION ANALYTICAL LABORATORY :I ,, To: Lisa Stevenson - (8-5568) - ET&SS - 2-3E-09 Request # GID:71638 From: Tom Kestner - (3-5633) - SMMD Analytical Lab - 236-2B-I 1 S,ubject: Date: Characterization of TCR-619 by IH-NMR and 19F-NMR Spectroscopy October 29, 2002 SAMPLE DESCRIPTION: TCR-619, lot U11N from the Telomer project. Nominal product = CF3(CF2),-CO2H, where average n = 9 (white powder). i s .a.mple Spectra#'s .. ExperimenDt escriptions i TCR-619 H71638.GID.404 400 MHzJH-NlVlRin acetone-drsolvent+ p-H.FXcrossintegration/internasltd. TCR-619 F71638.GID.405 376 MHzJgF.NMRin aectone-d_solvent+ p-HFXcrossintegration/internasltd. OBJECTIVE: This sample was subjected to a combination of IH-NMR and 19F-NMRspectral analyses to determine the purity of the nominal product. Special emphasis was also placed on attempting to identify and quantify any impurity components. EXPERIMENTAL: A portion of the sample was accurately weighed, spiked with a known amount of 1,4-bis(trifluoro- methyl)benzene (p-HFX), and then totally dissolved in deuterated acetone (acetone-dr) for subsequent analysis by NMR.. A 400 MHz IH-NMR spectrum and a 376 MHz WF-NMR spectrum were acquired at room temperature using a Varian UN1TYplus 400 FT-NMR spectrometer. This sample preparation method permitted the p-HFX to be used as either 1) a tH/19F-NMR intemal standard to allow the calculation of the _t absolute weight percent concentrations of specific components, or 2) a II-I/I_F-NMR cross integration standard to permit the cross correlation of the relative IH and 19Fsignal intensities for evaluation of the overall sample composition. _i_ RESULTS: The IH-N'IvIR and 19F-.-"q'IWII_ spectral data indicated this sample was a relatively high purJt-y forna o the nominal product, CF3(CF2),-CO2H, where the average value ofn = 8.63. Small amounts of a few impurity components, including probable isomers, were also assigned. A II-/./t9F-NMR cross integration analysis technique was then used to calculate the relative weight percent concentrations of the identified components. The qualitative and quantitative compositional results that were derived from the single trial _ ]H/19F-NMR cross integration analysis are summarized below in TABLE-1. The relative weight percent "" concentrations shown in TABLE-I should be very close to their respective absolute weight percent values assuming no water was present in the sample. Trace amounts of numerous other unassigned components were also detected in the 19F-NMR spectrum, but additional work would be needed in an effort to identify or quantify some of these other components. Copies of the NMR spectra are attached with the paper copy of this report for your reference. If you have any questions about these results, please let me know. - ExactCopyof Or/g_nal Pagel of 2 lnlUal Page 189 of 225 , i-?-.i_ I fJ '_ i i !=i,i_ i! i=.! -1 i_] !i:_ 3M ENVIRONMENTAL LABORA TORY E02-1039 October 29, 2002 3M SMMD Analytical Lab Request # GID:71638 TCR-619. Lot U11N: Telomer Project Tom Kestner e: Mark EIIcfson Rick Payfer Ron Purcell William Reagen Fil Reference: 1571638.GID.TCR-619_Lot U 11N.. Tdomcr Project.DOE/I OI TABLE-1 Sample: TCR-619. Lot U11N from the Telomer project. Overall Compositional Results by IH/tgF-NMR Cross Inte[_ration Analysis Component Structures' NMR R(esliantgilvee trWiael igmheta*s/u, reCmoenncte)ntrations CF3(CF2),-CO2H 96.4% , (where average n = 8.63) Probable internal branched isonzers 2.6% CF3(CF2)x-CF(CFj)-(CF2)c-CO_H . (where x;e0 and assume x+y = 6,63 for calculation purposes). Probable trans-olefins of general form: trans-CF3(CF2)x-CF=CF-(CF2)y-CO2H 0.89% (assume x+y = 6.63 for caleulat!on pl .U'l_.OS)eS Possible terminal isopropyl branched isomers (CF3_z-CF-(CFz),-CO2H (assume n=6.63 for calculation purposes) 0.12% Probable C.H2,.2 saturated aliphatic hydrocarbons Total inorganic fluoride 0,032% >0.0017% 1. Trace amounts of numerous other unassigned components were al_o detected in the 'gF-NMR spectna_ Page 2 of 2 ExacCt opyof 0riginal Page 190 of 225 3M _NVlRONMENTAL LABORATORY ,, E02-1039 i:i ,_,_! i i ! ..... ii ii i_J _ !t i] _:; !?_,_'_ ',_;_ !_ (:_ i,L.. i > tLJ Certificate of Analysis Nominal Product: CF3(CF2)n-COzH,where average n _ 9 Perfluoroundccanaociicd Product Code: TCR-619, Lot UI 1N October 29, 2002 The sampleofTCR-619, lotUIIN wasanalyzeudsinga combinatioonfZgF-NMR and IH-NMR _cctral analysitsechniquesT.he overalqlualitati,vaenqduantitaticvoempositionraelsulttshatwerederivefdrom thesecombinedanalyseasresummarizedbelowinTABLE-I. TABLE-1 Sample: TCR-619, Lot U1 IN , OverallCompositionaRlesultbsy Combined19FInH-NMRSpectraAlnalyses Component Structures i II.{/19F.NIk/IR " Relative Weight% Concentrations (sinGle trial analysis) . CF3(CF2).-CO2H (where average n = 8.63) Probable internal branched isomers CF3(CFz)x-CF( CF3)-(CF2)y-CO2H (where x_0 and assume x+y = 6.63 for calculation purposes) Probable trans-olefins of general form: trans-CF3(CF2)x-CF=CF-(CF2)y-CO2H (assume x+y = 6.63 for calculation purposes) Possible terminal isopropyl branched isomers <96.4% Purity 2.6% 0.89% 0.12% (assume (nC=F63.6)23-CfoFr-(CcaFlc2u)l.-aCtioOn2Hpurposes) Probable C.I'-I2.+2 saturated aliphatie hydrocarbons O.O32O/Jo'" "' Total inorganic fluoride >_0.0017% 1. Trace amounts of numerous other unassigned components were also detected in ttm '9F-NMK speettl, lrcL, -,,. Tom Kestner ExactCopyof Original oate Page 1 of I File Ref:rence: CofA_TCR-619_Lot UI IN.doe Page 191 of 225 ";::.i ":_,:_ "_=:_t' i : "' ::; _ i i_!l :::! _ :'_J :i _#;,_ "_ , r,l,_MEIV_fC_NMEZVL_AaOP_T,O4 ffY OaLkwoodProducts, Inc. 1741 Old Dunbar Road West Columbia, SC 29172 Phone(803)7"39-8800 :. Fax(8031739-6957 CERTIFICATE OF ANALYSIS Da{e: 15-Jui-02 .. Material: Perfluoroundecanoic acid : Cat.No.: 2265 ' Cas Na.: Lot No.: [4234-73-5] UIlN Assay: 99+%by NaOH titration Appeaxance; _hlte solid Melting Point: 92-98_C ; :,;.:. ."_, ,; ,= ..._. .. ..,, :. - ' : ":.,'. : "_ ..:,.::. :_. "-7". _:-2:.,. .-%, .... : .:: .,: :_. .....: ;.!. ":))':'": .!! "' ". 'e @ . .:.:. . "";;.: ."g ........ :_ .-;:,: . _..... '" ...".. ,:: .- .j '. ". :.:,: ._" Exactopoyfoa_al .,'.' _ .;_ _ _/'-- :.: _: --.. ..; ., "t, :._ "" '_i -, .._. .;..!!:.'. ,, .. .._ .... ', . .,.. . ';':"-"'":" .i ;. .i Page 192 of 225 3M ENVIRONMENTALLABORATORY ', ' E02-1039 USE LOG J ...... l_erfluoroundecanoicacid ; PrCeRrf-l6u1o9roundecanolcacid I -_ I Before withdrawn after or reasonfor removal) withdrawal (massorvol) withdrawal I BalanceIO 1 BalanceID 2 BalanceID 1 J15.S264g 10.0691g ]15.4573g _standard02040-65 [914 I" IRWW,. . 108/29/2002 I /'i .J L i '_./,i IniU_. Dato .... J Page 193 of 225 ,..... ': i' :.... 3M ENVIRONMENTALlABORATORY E02-1039 Data: Analyst" SINGLE COMPONENT PREPARATION LOG 2..- /__ . RookNo. 02 040 Description: ft'e//..,,-b/.t.J/.,.,c ._;J )I Weight or Volume " Used: <_ _'* _,_ .. BalanceIO: ._ I C/ Concentrationor purity: __,, /,//4 o_ecr o=aeon Faro,,: ,_ ,d- Solventand TN.A J Storage Location: _,_ _) F-_, ,_ ,_-/_ / <_ I ExpirationDate: ,_ ___/OJ_ , ExacCt opyofOriginal gnatum. ' Date" Page 194 of 225 ii _"_ _,'_ '_,_ _'_/ ., ' _: ! .i ,-._ "l _.J :: _i_ 3M ENVIRONMENTALLABORATORY E02-1039 Test Controland ReferenceSubstanceLog Substancetradename orreference# Substance/chemical name: ILoUbatch#: 13M# Perfluorododecanoicacid Perfluorododecanoiacid R24K [D#2266 TCR Substance# TCR # Receivedfrom: SD037 TCR-37 OakwoodProducts Inc 4/2/99 'Expirationdate: Jlnitials: Number/sizeof containers: ICondition: IRetain 12/01/2010 JCP II120 mL plasticbottle white powder MCH07/22/99 ).2159g vol):Amreucnetived(wt.or 25g, 43.4548 g gross wlL Date: 04/2711999 Shipper:. NIA MSDS (y/n) Dateof Retain lid y (._)N 1011811999 J_; .)!Ar cfiiVed/Subs{a:nceiN0t.Available_l Purity: 99.65% updated 1013010O2 K, NMRanalysis Recordsreceived: MSDS,Certificateof Analysis Locationof synthesis, Unknown ' fabrication,or derivation records: Std Location/Storage: iF19, Frozen MCoomlemcuelnatrFsormula: CTNl 1-AF.2234C5O3 pOrHiorto 4/27/99 JCP 04/27/99 Standardhas been movedto Freezer19 in room 347 KJD 06/06/00 Standardwas storedat room temperaturepriorto 06/06/00. LAC 12/19100 Shippingcomments: Irritant, no shipping information- ship as small quantity exception. LAS 10102/02 Attachment(s) _ sd037msds.pdf _ sd037cofa.pdf L_ NMR SD037.pdf ;_i..., ExacCt opyof Original _ Da_ Page 195 of 225 3M ENVIRONMENTALLABORATORY E02-1039 _,_ i "'-:_i I 7 ! !.:','!'ii iiii_,i iI ::i,_ : !_ili I __r !_i _ i I i _":_i__.. 3M SPECIALTY MATERIALS MANUFACTURING DIVISION ANALYTICAL LABORATORY To: Lisa Stevenson - (8-5568) - ET&SS - 2-3E-09 Request # GID:71638 From: Tom Kestner - (3-5633) - SMMD Analytical Lab - 236-2B-11 Subieet: Date: Characterization of SD037 by tH-NMR and tgF-NMR Spectroscopy October 28, 2002 SAMPLE DESCRIPTION: SD-037, lot R24K from the Telomerproject. Nominal product = CF3(CF2),-CO2H, where avera[e n = 10 (white powder). I Samples Spectra#'s ExperimenDt escriptions SD037 SD037 HT1638.GID.405 400 MHz'H-NMRin aeetone--dsnolvent+p-HFXcrossintel]xation/internsatld. F71638.GID.406 376MHz 19F-NM_ in acetone-dosolvent+ p-HFXcrossintegration/internasltd. OBJECTIVE: This sample was subjected to a combination of tH-NMR and tgF-NMP,,spectral analyses to determine the purity of the nominal product. Special emp.hasis was also placed on attempting to identify and quantify any impurity components. EXPERIMENTAL: A portion of the sample was accurately weighed, spiked with a known amount of 1,4-bis(trifluoromethyl)benzene (p-HFX), and then totally di,;solved in deuterated acetone (acetone-dr) for subsequent analysis by NMR. A 400 MHz _H-NMR spectrum and a 376 MHz 19F-NMR spectrum were acquired at room temperature using a Varian UNITYplus 400 FT-NMR spectrometer. This sample preparation method permitted the p-HFX to be used as either 1) a IH]IgF-NMR internal standard to allow the calculation of the absolute weight percent concentrations of specific components, or 2) a tH/tgF-NMR cross integration standard to permit the cross correlation of the relative IH and 19Fsignal intensities for evaluation of the overall sample composition. RESULTS: The |H-N'MR and 19F-N1MIR spectral data indicated this sample was a high purity form of the nominal product, CF3(CF2)a-C02H, where the averag0 value ofn = 10.042. Small amounts of a few impurity components, including probable isomers, were also assigned. A tH/t 9F-NI_L-Rcr.oss integration analysis technique was then used to calculate the relative weight percent concentrations of the identified components. The qualitative and quantitative compositional results that were derived from the single trial _.-.. _H/_'_F-NMRcross integration analysis are summarized below in TABLEd. The relative weight percent concentrations shown in TABLEd should be very close to their respective absolute weight percent values assuming no water was present in the sample. Trace amounts of a few other unassigned components were also detected in the NMR spectra, but additional work would be needed in an effort to identify or quantify these other components. Copies of the NMR spectra are attached with the paper copy of this report for your reference. If you have any questions about these results, please let rae know. ExactCopyof Odg[nal Page1 of 2 Dat.a Page 196 of 225 3M ENVIRONMENTALLABORATORY E02-1039 _ October 28, 2002 3M SMMD Analytical Lab Request # GID:71638 SD037, Lot R24K: Telomer Project '_ Tom Kestner e: RickPayfer Ron PurcEll WilliamRezgen FileRcfenmceb:71638.GID.SDO37.R_I2._4KTlomcPrrajeetDOCllOI TABLE-1 Sample: SD037, Lot R24K from the Telomer project. Overall Compositional Results by 1I-{/19F-NMRCross Integration Analysis Component Structures i NMR Relative Weight% Concentrations ...i.,. (single trial measurement) ':ii CF3(CF2)n-CO2H 99.65% where average n = 10.02 "_ Probable (CF3)2-CF=(CFz)n-CO2H O.13% 'I .; assume n=8 for calculation purposes Possible CF3(CFz)x-CF(CF_)-(CF2)y-COzH 0.088% : where x_0, _t-_0and assume x+y = 8 for calculation purposes Probable chlorinated impurity, CI-CF2CF2-Rt; 0.049% possibly as CI-(CF2)II-CO2H Possible methyl ester impurity as 0.048% CF3(CF2)n-CO2CH3 where average n = 10.02 Probable C.H2.+2 saturated aliphatie hydrocarbons 0.016% iii' 1. Trace amounts of other unasTsiognlueednceomponents werealso detected in the NMR spectra. 0.015% t ExactCopyofOriginal Page 2 of 2 Page 197 of 225 3M ENVIRONMENTALLABORATORY E02-1039 ..._. ' ,_ ,t _i_i:,l -I I -" ::i .,....J '__ !_ :i " i I [ C erti [icateo fA na lysis Nominal Product: CF_(CF2).-CO2H, where average- n _ 10 perfluorododecanaociicd ProductCode:SD037,LotR24K October28,2002 Tom Kestner The sample of SD037, lot R24K was analyzed using a combination of 19F-NMR and IH-NMR spectral analysis techniques. The overall qualitative and quantitative compositional results that were derived from these combined analyses are summarized below in TABLE-1. TABLE-1 ' Sample: SD037, LotR24K Quantitative Compositional Results by Combined 19F/tH-NMR Spectral Analyses Component Structures _ _H/19F-NMR Relative Weight% Concentrations (single trial analysis) CF3(CP2),-CO2H whereaveragen = 10.02 Probable(CF3)2-CF-(CFz),-CO2H assumen=8 forcalculatipounrposes Possible CF3(CF_)x-CF(CFj)-(CF2):CO2H where x;t0, y_0 and assume x+y = 8 for calculation purposes Probable chlorinated impurity, CI-CF2CF2-Rf, possibly as CI-(CF2)H-CO2H Possible methyl ester impurity as CF'_(CF2)n-COzCH3 <99.65% Purity 0.13% 0.088% 0.049% 0.048% where averase n = 10.02 Probable C,,H2,+2saturated aliphatic hydrocarbons 0.016% Toluene 1 0.015% 1. Trace amountsof otherunassignedcomponentswerealsodetectedin theNMRspectra. %. ExactCopyof0rfg[nal Page I of I FileReference:CofA_SD037_LoRt24K.doc Page 198 of 225 i.! !.'._.I ." . _:'_ t :i i i_J f_:i 'J '_ _._, ., t._J , 1 %,, OAKWOOPDRODUCTS Fax:803-739--6957 Feb _/t I_JRON_I_ri'AL I...AfJ_'t_TORY _ Oa]_vood]Pxod_ts, Inc. 17_1OldDunba_ Road West Columbia, $C 29172 Phont (S0_)739-eeoo Fax(8o3)_39-_957 CERTIFICATE OF ANALYSIS Dat_ 11-FeIMJ0 i Materia]: Perflaorododecanoic add J i Cat.No.: 2266 " Lot No.: Assay: R22K J Products more volatile than Ferfluorododecanoic acid < 2% Feffluorododecar_ acid 196%rain. l_roducts less volatile than Peffluorododecanoic acid' <2% Appearance: WRite solid Melting Point: 107-I09C ! ! ExacCt opoyfOriginal i _uality C6_ffol _" Page 199 of 225 3M dNVlRONMENTAL LABORATORY E02-I039 : I useLOQ 1 !_i _erfluorododecanoicacid ...:._! _I$eDrf0lu3o7 rododecanolc acid 1 I j Before withdrawn after or reasonfor removal) :-I I withdrawal (massor vol) withdrawal : IGBraolsasnWcetI.DNo1l. I BalAanmcnetI.D 2 IGBroaslasnwcte/IvDol1l Pulpose(enterstandardnumber o,,, 142.5752g 10.0796g '' 142.4956g. tstandard02040-55 _914 _. IRVVw .,,108/29/2002 l } := _:._._ I!!';_!,! ExacCt opoyfOriginal _al Date Page 200 of 225 3M ENVIRONMENTALLABORATORY E02-1039 Analyst: //4_A,_ Page No. 55 StockNumber:. _",/_ _.J_ .. i Weight orVoluma _._' Used: _ _:p,..,r"_,,_'_ J BalanceIO: _/q Pu,_: /i_ Concentrationor :.:_ i:?! Cor_c_Wdeig._ OtherCorrection Fa_,',,: /V/'" Solventand TN-A _ FinaVl olume: ,__'___ ] FinaCl oncen_tlon: / / 2._ / . .L ! ' Storage Location: 2.J _/'_ --" t/_,- ,_'" --_ ExpiratlonDate: ,_/_?/0 ,._> .... ExacCt opoyf 0dglnal _tV_(. ,/,/<_",-- : InlBl _lgna_e," DSta - Page 201 of 225 . 3M ENVIRONMENTAL LABORATORY E02.1039 i Test Control and Reference Substance Log Substance trade name PFOS TCR Substance # SD018 :i or reference # ' Substance/chemical Potassium Perfluorooctane I"CR # rCR-18 name: Sulfonate FC-95 Lot/batch #: 217 IReceived from: lugDickes 8/10198 I 3M # date: 98-0211-0888-S I,,morouonto(w,vo,eI.Ir'd"O.Oo'rOo'.g II VO ): I I Initials: PMR IDate: 104107/1999 I containers:Nuomfber/size t/175 mL glass container ijShipper: !lUnknwn I ICondition: white powder MCH-07122/99 IMSDS (y/n) _ll Y (.) N I Retain 0.1733g tDate of Retain 110118/1999 1 /_i ?::' ' i i _' - ("_ ArchivedlSubstance NotA_ailable_J Pudty: 86.9% LAG 09119100 IRecords received: I NMR Results/Charectedzation #53030, MSDS KJD 02/29100 Interim final report from Centre and Certificate of Analysis. LAG 04126/01 [Location of synthesis, Ifabfication, or derivation Irecords: 98-0211-0888-5 _Std LocatJonlStorage: Molecular Formula: Comments F19, Frozen C8F17SO3-K+ Environmental Lab Traceablitity numberwas TN-A-2130 PMR 04/07199 Moved to cold storage (< OC) on 05116/00 LAC 05116/00 Standard has been moved to Freezer 19 in room 347 KJD 06/06100 Standard was stored at room temperature prior to 05116100. LAC 12/19/00 Shipping Codes: FC-S000-0185-6(<30 g on dry Ice) LAC 06/14101 Shipping Comments: Dangerous goods in excepted quantity of Class 6, UN281t. REGULATED-TOXIC 98-0211-0888-5 (>30. g (1 oz sample)) Not on dry ice. LAG 01/08/02 sdO18msds.pdf sdO18nmrreq53030.pdf sdO18nrnrreq61517.p( ,v:,,_, E00-1682-COA-SD018-PFOS-Rev3 ExactCopyof Original Page 202 of 225 ! _'I '_:-_i ' ii :,....J I:_ '_ '_1 ..... !_i! ' _! _!_, :'; _ i r k: :__JI 3M ENVIRONMENTALLABORATORY E02-1039 _lkV._'_'_k'_mli. '_P'O_'_ll=._Phone: Cntr Analgtical Laboratories, Inc. 3048 ResearchDrive State College, PA 16801 www.centrelab.com (814)231-8032 Fax:(814) 231-1253 or (814) 231-1580 INTERIM CERTIFICATE OF ANAL YSIS Revision 3 Centre Analytical Laboratories COA Reference 3M Product: PFOS, Lot 217 Reference #: SD-018 #: 023-018A Test Name Purity I Purity: 86.9% Specifications i i Result 86.9% Appearance Identification NMR Metals (ICPfMS) I. Calcium 2. Magnesium 3. Sodium 4. Potassium 2 White Crystalline Powder' Conforms Positive I.O.O05wt./wt.% 2. 0.001 wtJwt.% 3. 1.439wtJwt.% 4. 6.849wt./wt.% 5. Nickel 6. Iron 7. Manganese Total % Impurity (NMR) Total % Impurity (LC/MS) Total % Impurity (GC/MS) Related CompoundsPOAA Residual Solvents (TGA). 56.. <O0..O00015wwt.t/.w/wt.t%.% 7. <0.00lwt./wt.% 1.91 wt./wt.% 8.41 wt./wt.% None Detected 0.33 wt./wt.% None Detected PInuorirtgyanbicy DASniCons (IC) 1. Chloride 2. Fluoride 3. Bromide 4. Nitrate 5. Nitrite 6. Phosphate 7. Sulfate 4 Organic Acids _(IC) Not Applicable _ 1. <O.O15 wt./wt.% 2. 0.59 wt./wt.% 3. <0.040wtJwt.% 4. <O.O09wtJwt.% 5. <0.006wt./wt.% 6. <0.007 wt./wt.% 7. 8.76 wt./wt.% 2. PFPA 3I. THFFABA 4. NFPA Elemental Analysis*: 1. Carbon 2. Hydrogen 3. Nitrogen 4. Sulfur 5. Fluorine 1. Theoretical Value = 17.8% 2. Theoretical Value = 0% 3. Theoretical Value = 0% 4. Theoretical Value = 5.95% 5. Theoretical Value = 60% 2. <0.1 wt./wt.% 31.. <0.01.01wwt./tw./wt.%t.% 4. 0.28wt./wt.% l. 12.48 wt./wt.% 2. 0.244 wt./wt.% 3. 1.74 wt./wt.% 4. 8.84 wt./wt.% 5. 54.1 wtJwt.% __ =_ v, " o u__ _l COA023-018A Page I of 3 Page 203 of 225 "L'." 3M ENVIRONMENTALLABORATORY i E02-1039 ...i.: _la_k_'_i_ Centre Analgtical Laboratories, Inc. 3048 Research Drive State College, PA 16801 www.centrelab.com '_e'O"_._Phone: (814)231-8032 Fax:(814)231-1253or(814)231-1580 ,'_ INTERIM CERTIFICATE OF ANAL YSIS "_i Revision 3 Centre Analytical Laboratories COA Reference #: 023-018A Date of Last Analysis: 08/31/00 Expiration Date: 08/31/06 Storage Conditions: Frozen _<-10C ._ Re-assessment Date: 08/31/06 . _,_ IPurity = 100% - (sum of metal impurities, 1.45%+LC/MS impurities,8.41%+Inorganic i Fluoride, 0.59%+NMR impurities, 1.905%+organic acid impurities, 0.38%+POAA, 0.33%) Total impurity from all tests = 13.07% Purity = 100%- 13.07% = 86.9% 2potassium is expected in this salt form and is therefore not considered an impurity. 3Purity by DSC is generally not applicable to materials of low purity. No endotherm was _ observed for this sample. 4Sulfur in the sample appears to be converted to SO4 and hence detected using the inorganic anion method conditions. The anion result agrees well with the sulfur determination in the elemental analysis, lending confidence to this interpretation. Based on the results, the S04 is not considered an impurity. .,, I-IFBA Heptafluorobutyric acid NFPA Nonafluoropentanoic acid PFPA Pentafluoropropanoic acid 6Theoretical value calculations based on the empirical formula, CsFt7SO_'K+(MW=538) __ ; Tlfis work was conducted under EPA Good Laboratory Practice Standards (40 CFR 160). :ii COA023-018A Page 2 of 3 Page 204 of 225 ' i ii'!:,.l i:iill , :] :J i IVA'_'_'_l '_'O"_k'_k'_i_e 3M ENVIRONMENTALLABORATORY E02- f 039 CEntPGAnalytical Laboratories, Inc. k ' 3048 Research Drive State College, PA 16801 www.centrelab.com hone: (814) 231-8032 Fax: (814) 231-1253 or {814) 231-1580 INTERIM CER TIFICA TE OF ANAL }'SIS Revision 3 Centre Analytical Laboratories COA Reference #: 023-018A LC/MS Purity Profile: Impurity C4 C5 C6 C7 Total wt./wt. % 1.22 1.33 4.72 1.14 8.41 Note: The C4 and C6 values were calculated using the C4 and C6 standard calibration curveu, respectively. The C5 value was calculated using the average result from the C4 and C6 standard curves. Likewise, the C7 value was calculated using the average result from the C6 and C8 standard curves. Prepared By: /o _,/o/ _ Charles SimO_ D "_ , Scientist, CentreAnalytical Laboratories ._. o , Reviewed By:/_hn Flaherty / Date Laboratory Manager, Centre Analytical Laboratories COA023-018A Page 3 of 3 Page 205 of 225 3M ENVIRONMENTAL LABORATORY E02-1039 J USELOG FOS ....] Fotasslum Perfluorooctane Sulfonate FC-95 -_i _SD018 Before withdrawn after or reason for removal) withdrawal (mass or vol) withdrawal BraolasnscWet.INOol1. I BalaAnmcneLIO 2 I BGarolasnscweUIDvo1l I Purpose (enter standard number I Bal_nce I Ball_nce J Initials I Date I 1140.44eOg [0.0814g [140.36680 Istandar0d2022-56 ]914 I" [RWW 10,5/13/2002 I i L,,J _.:,'.;:,_ 21 i E;;;.:tCopoyfOtfg/_ Page 206 of 225 ,::!4 .::I.LI.; ... 3M ENVIRONMENTAL LABORATORY E02-1039 Date: Analyst: SINGLE COMPONENTPREPARATIONLOG ,_., #(_112A) pp >J .s'b<.g Book _o. 02 022 Page No. 56 ;" _ ;"3]! ,i I_,_ *'_ ,,_r_!'.__:_.,,... StockNumber. ,.5/) 0/_ u,=t: Weight orVolume /__..o_ Concentratioonr Pu_s:. El, 9 _ CorrectedWeigh_ _"_, J_ r,,,= #" B=,..Ico.: ?/ OtherCorrection Factora/:), 92 7J- Solvent aNndumTbNe-rA. /6_? _,ff/._'A/,;-A -_OZoJ'J/ FinalVolume:_ _-0_-. / :. ;j :,; Slorage Location: /i_ / _, ',] RmllConcentration: l _DI./_',_/_,, ...... ExplmUon Date: ( [k3/_ J." ExactCo_yof Original Initial Dats 5_r_llum; Dil Page 207 of 225 _:;::_ k,i i ATTACHMENET: PROTOCOL AND AMENDMENT 3M ENVIRONMENTALLABORATORY E02-1039 :_: :21 Page 208 of 225 i 3M ENVIRONMENTALLABORATORY E02-1039 t :_i! i_Jii-!:i _ i-I :! _, i STUDY PROTOCOL STUDY TITLE Analysisof EndogcalouFsluomchcmicailns NormalPooledHumanSerumandPlasma SPONSOR William K, Reagen,Ph.D DATA REQUIREMENT 4.0CFRPart792 TESTING FACILITY 3M EnBvuirioldnimngen2-t3aElL-a0b9oratory 935 Bush Avenue St Paul, MN 55106 LABORATORYSTUDY IDENTIFICATION 3M Environmental StudyNumber E02-1039 NUMBER OF PAGES 10 ExacCt opyofOdginal Page 209 of 225 i '_ '_ ,!_I ii_ i, ....:. : Anatyslso( EndogenousRuotochemicaltt 3M ENVIRONMENTAL LABORATORY E02-1039 E02-1039 STUDY IDENTIFICATION Analysis of Endogenous Fluorochemieals in Normal Pooled Human Serum and Plasma SPONSOR Srt1_rDh_CrOR TEst FAazdza, William Reagen, Ph.D. 3M Environmental Laboratory 935 Bush Avenue, Building 2-3E-09 St. Paul, MN 55106 (651)778-6565 Mark E. Ellefson 3M Environmental Laboratory 935 Bush Avenue, Building 2-3E-09 St.Paul,MN 55106 (651)778-5405 3M Environmental Laboratory 935 Bush Avenue, Building 2-3E-09 St. Paul, MN 55106 Experimental StarDtate 10 October 2002 _ Experimental Termination Date 01 November 2002 i %. 3M EnvironmentaL|abotatt3P/ ExactCopyof Original Page2 of 10 " Page 210 of 225 3M ENVIRONMENTALLABORATORY E02-1039 "L I AnalysoisfEndogenoFulsuoroctlemicale E02-1039 ':_, L.: iJ ' 1.0 Introduction and Purpose/Objective 1.1 The purpose of this study is to quantify perflunrohexanoic acid (C6), perfluoroheptanoic acid (C7), pentadecafluorooctanoic acid (C8), heptadecafluorononanoic acid (C9), nonadecafluorodecanoic acid (CI 0), perfluorodecanoic acid (CI 1), perfluorododecanoic acid (C12), tetrahydroperfluorooctan sulfonate (THPFOS), tetrahydroperfluorodecane sulfonate (THPFDS), and perfluorooctanesutfonate (PFOS) in normal pooled human serum and plasma. This study does not have a typical test substance that is dosed onto a specific controlled test system as per a conventional GLP study. 2.0 Regulatory Compliance 2.1 This study will be conducted in accordance with the United States Environmental Protection Agency Good Laboratory Practice Regulations for the Toxic i] Substances Control Act, 40 CFR 792. ' 3.0 Test Substances 3. l Preliminary screening indicates that the test compounds listed below are present as endogenous material in normal pooled human serum and plasma. Information pertaining to traceability, source, physical description, and storage conditions is not available for these compounds as they exist in biological matrices. _,:_ Test Substances Tee!Substance ,:_, PerfluorohexanAocicid t,_ , ] TetradecafluoroheptAancliodc PentadecafluorooctaAncoiidc ;'_!:, -_ ": " HeptadecafluotononaAncoidlc NonadecafluorodecaAncoidlc PerfluoroundecaAncoildc PerfluomdodecaAnocid 1H,2H,3H4,H-perfluorooctanesulfonate 1H,2H.3H.4H-per_uomdescualnfoenata i PotassiumPerfluorooctanesulfonata Formula CsF.COOH ,, CaFI=COOH CTF_=COOH C=F_rCOOH CoFI_=CGOH CloF_tCOOH CuFz=COOH C=H=FI=O=S C_oH=F_IO=S C=F_TSO_'K : ExacCt opyof Original 3MEnvironmeLnatabloraton/ Page3 of10 Page 211 of 225 ', j Analysisof EndogenouaFluorochemicals 3M ENVIRONMENTAL LABORATORY E02-1039 ". E02-1039 4.0 Reference Substances "!_ii i i :_.:_ ! :! !_ ',_',! __i''_ Reference Substances SRuebfesrteannccee For.mula Trace#ability Source Perfluorohexanoic Acid Tet dec no, heptanioc AcJd Pentadecafluom octanoic Ar.Jd Heptadecafluoro nonanoic AcJd C=F_ICOOH TCR-047 SMM* 236-1B-10 c,,COOH TCR-267 ,,,,, C_FnCOOH TCR-617 Oakwood Products C.F_K_OOH TCR-618 Oakwood Products NdoencaadneocicafAlucoidro C_._COOH TCR-038 OParokdwuocotds Perfluomun decanoic Acid C,_=_COOH TCR-619 Oakwood Products dPecearfnluooicroAdcoid 1H,2H,3H,4Hperfluorooctane sulfonate Ct_Fz._COOH ,,,, TCR-037 ,, , C=%FI=O=S TCR-343 OParkowduocotds SynQuest Labs 1H,2H,3H,4Hperfluorodecene sulfonate C_0H=F_70=S TCR-627 Pace Analytical Potassium Perfluomoctane sulfonate C,F_SO='K *Documcatution of the method ofsynthc_is TCR-018 SMM* 236-1B-10 hi located at the source. DPehscyrsipictaiol n ,, J,,,,, Colorless Liquid iear Crystals White Crystals White Crystals White Solid Purity TBD** 99.5% 97% > 99% 98% White Crystals PWowhidteer ,, > 99% 96% White Powder TBD White Crystals White Powder 94.7% 86,9% CSontodriatigoens Frozen Frozen Ambient Temperature Ambient Temperature Frozen Ambient Temperature Frozen Frozen Ambient Temperature Frozen **A sample of the reference substance ha=been sent for characterization ':':_ 5.0 Test System i _. Test Systems Pooled Human Serum Sigma-Aldrich, Milwaukee, WI TCR-689 i Pooled Human Serum Lampire Biological Laboratories, TCR-688 :: Pipersville, PA Peered Human Serum Bioresource Technology, Inc., Fort TCR-687 Exact Copyof Original : Lauderdale, FL _ IO3_ =, Pooled Human Serum CA TCR-690 ---- 3M Environment_Laboratory Page4 of 10 . Page 212 of 225 i Analysis of Endo_genous Fluorochemical.s :.=., 3M ENVIRONMENTALLABORATORY E02-1039 E02-1039 . :._, ,_ I :i _ "_ 1!;_,I_ I_+ .__ ....... . PooledHumanPlasma PooledHumanPlasma LampireBiologicaLlaboratories, PipersviUeP,A GoldenWestBioCloAgicalsT,emecula, PooledHumanPlasma _ InnovativRe eseaMrIchI,nc.iSouthleld, PooledHumanPlasma CentralChina TCR-685 TCR-684 TCR-683 TCR-674 5.1 Justification of the test system. Based on preliminary testing, normal pooled serumand plasma contain endogenous levels of thetest analytes. 5.2 Identification of Test System, Samples shall be identified by the study number, date of initial preparation, test substance or test system, sanaple number, replicate number (if applicable), analyst(s), and project leader. 6.0 Surrogate Matrix Surrogate Matrix RabbitSerum J Sigma-Aldrich TCR-686 6.1 Justification of the Surrogate Matrix. Based on preliminary testing, normal pisoothleedrebryabsbuititasebrluemforcounsteaianss avseuryrrologwateemndaotrgixen. ous levels of the test analytes and 6.2 Induemntbifeirc,atidoante oof finSuitriarol gpareteparMataiotrnix, . tesStasmupblsetsanschealolrbseuirdroegnatitfeiedmabtryixt,hesasmtupdlye number, replicate number (if applicable), analyst(s), and project le'ader. "_,. 7.0 Analytical Methods 7.1 Analysis of the test and reference substances will be conducted in the 3M Environmental Laboratory. The analyses will be conducted as described by 3M Environmental Laboratory Method ETS-8-231, "Solid Phase Extraction and Analysis of Fluorochemical Compounds from Biological matrices", ExactCopyof Original Initial gala 3MEnvironmeLnatabloratory Page5of10 Page 213 of 225 Analysis of Endogenous Fluorochemicals 3M _'NVIRONMENTALLABORATORY E02-1039 E02-1039 !i:_,i i:r !=_:'! , i.il.i ii_'_;! _r"_ _'_:i i..; ,_ "'. i _' i !iiiii 8.0 Data Quality Objectives 8. I Absolute Recovery: 8. I. I The absolute recovery of the method will be evaluated separately in human serum and plasma and rat serum. For each matrix, the samples will be fortified at two levels of(500ppt and 5ppb). All samples will be extracted through the method, and analyzed by comparison with external calibration of non-extracted standards. The non-extracted standard calibraticounrveswillbe prepareidnmethanola,ndwillconsisotf a minimum ofnine(9)levelsi,ncludinagmethanolblank.The best appropriarteegressiwoinllbe usedtodescribtheis curve(forbest accuracyatalllevelosfthestandards). 8.1.2 The accurac(y% recoverya)ndprecisio(n%CV oftherecoveriewsi)llbe deterrninactdeachlevela,ndforalllevelcsombined.Thereisno control limiftoraccuracyp;recisiomnustbebettetrhan15% ateachlevel. 8.2 CalibratioCna:libraticounrveswillbepreparefdromextractemdatrixstandards, inChineseplasmaandrabbisterum.The curveswillconsisotfaminimum of nine(9)levelasnda matrixblank.The equatiownillbe determinebdy regression analysiussingthepeakareasoftheanalyteT.he accuracoyfeachlevelwillbe vcrificdA.ny leveloutsid7e5% -125% ofnominalmustbe deactivateadn,d regressiorne-calculateexdc,epttheLLOQ whichmustbe within30% ofnominal. Alllevelmsustshow a responsgereatetrhantwicethatoftheblank.A maximum of four (4) levels may be deactivated in any one set, or the set will be re-analyzed. 8.3 Limits of Quantitation (LOQ): The lower limit of quantitation (LLOQ) will be determined for each analyte. The level determined as the LLOQ must show a recovery within 75% - 125% for the analytes and must show a response greater than twice that of the blank. These limits will be determined during the course of the study and documented in the raw data. Should the LLOQ level calibration be de-activated in a particular set, the practical limit of quantitation for this set will be raised to the next acceptable level. Samples below the practical LOQ of that set will be reanalyzed until quantitated in a set including the validated LLOQ. The upper limit ofquantitation (ULOQ) will be determined in serum and plasma. The level determined as the ULOQ must show a recovery within 75% - 125% for the analytes. These limits will be determined during the course of the study and documented in the raw data. Should the ULOQ level calibration standard be deactivated in a particular set, the practical limit of quantitation for this set will be lowered to the next highest acceptable level. Arty sample with an area greater than 110% of the highest acceptable standard will need to be diluted into the range of the calibration curve. If samples are diluted into the range of the curve during analyses and enough sample remains, a post-ran dilution validation will be performed to verify sample values. To Exact Copyo1'Orlglnal (t/_( _ i_ 3M Environmental Labotaton/ Page 6 of 10 . Page 214 of 225 ;; ,4 :,, L,ji<l _ :i':i :,_:i i :,j _-,, i! :i > : ...... .,.:._..::,i! ! AnalysoisfEndogenoFulusorochemicals :.= 3M ENVIRONMENTALLABORATORY E02-1039 E02-1039 perform the dilution validation, one sample will be separated into two representative samples (i.e. two I mL aliquots for fluid samples or two I gram amounts for tissue samples) then diluted using two procedures. The first procedure consists of diluting the sample with additional matrix prior to extraction (sere adding sera), while the second procedure consists of diluting the extract with solvent post-extraction (methanol extract adding additional methanol solvent). If the values are not within 15% of each other additional testing will be required to determine which value is a correct representation of the sample concentration. 8.4 Use o f Confirmatory. Methods Confirmatory methods are typica!ly not needed with LC/MSIMS analysis. 8.5 Demonstration of Specificity 8.5.1 The identification ofanalytes will be substantiated by chromatographic retention time, by the characteristic primary ion, the characteristic product ion, and isomeric proportions (where applicable). 7.6 Controi of Bias Two levels of matrix fortifications, prepared at known concentrations of the test substance and bracketing the anticipated range of the method will be evaluated to determine recovery and to evaluate method pcrformance. P.eagent and matrix blanks will be run with each set to evaluate the level of background interferences. 9.0 Statistical Methods and Calculations 9. l Statistical methods for the analytical results will be limited to the calculations of means, standard deviations, and relative standard deviations (as appropriate). 10.0 Report A report of the results of the study will be prepared by 3M Environmental Laboratory. " The report will include, but not be limited to, the following, when applicable: 10.1.1 Name and address of the facilities performing the study, 10.1.2 Dates upon which the study was initiated and completed. 10.1.3 A statement of compl:iance by the Study Director addressing any exceptions to Good Laboratory Practice Standards. 10.I.4 A copy of the protocol, and any amendments and deviations. 10.1.5 A description of the methods used to conduct the test(s). The report will ExactCopyof Original [ tw_ ,.i._,,l_ _/owg..._.L_u. _ 0at8 3MEnvitonmenLtaabJoralory Page7 of10 Page 215 of 225 '' i ' :_ ! : ::,_! _.... :',_I:_ :."_ ' I _ .._.!.. If_ )',:_, ,,_1 !_ ,_ _ ,o._ ;_!_ i_:_)_3 i /_alysis of Endc-,9_nousFluorochemicals 3M ENVIRONMENTALLABORATORY E02-1039 E02-1039 contain updated methods incorporating any changes or improvements. 10.1.6 A description of the test system. 10.1.7 A description of any circumstances that may have affected the quality or the integrity of the data. 10.1.8 The name of the Study Director and the names of other scientists, professionals, and supervisory personnel involved in the study. 10.1.9 A description of the transformations, calculations, or operations performed on the data, a summary and analysis of the analytical chemistry data, and a statement of the conclusions drawn from the analy ses. 10.I. 10Statistical methods used to evaluate the data, if applicable. I 0. I. IIThe signed and dated reports of each of the individual scientists or other professionals involved in the study, if applicable. 10.1.12The location where raw data and the final report are to be, stored. 10.1.13A statement prepared by the quality assurance unit listing the dates that study inspections and audits were made and the dates of any findings reported to the Study Director and Management. 10.2 If it is necessary to make corrections or additions to a final report after it has been accepted, the changes will be made in the form of an amendment issuedby the Study Director. The amendment will clearly identify the part of the final report that is being amended, provide the reasons for the amendment, and will be signed by the Study Director. 11.0 Quality Assurance 11.1 The 3M Environmental Laboratory Quality Assurance Unit will review the protocol and audit study conduct, data, and the final report to determine compliance with Good Laboratory Practice Standards and with 3M Environmental Laboratory Standard Operating Procedures. The Quality Assurance Unit will report all findings to the Sponsor Representative and the Study Director. t2.0 Location of Raw Data, Records, and Final Report 12.1 Original data or copies thereof, will be available at 3M Environmental Laboratory. When the final report is completed all original paper data, including those items listed below, will be retained in the archives of 3M Environmental Laboratory following signing of the final report. 12.2 The following raw data and records will be retained in the study folder in the archives according to 3M Environmental Laboratory SOPs. 12.2. I Approved protocol and amendments ExacCt opyofOriginal _ 3M EnvironmentalLabaralc=ty Page8 of 10 Page 216 of 225 _;-:.] ! ,:_ ::,_ ' _'_ _I ' "I i,..J __L'_:_.iit '/_:_j t'?,: _t I_ ""% AnalyaoisfEndogenoFulsuorochamlcals 3M _NVIRONMENTALLABORATORY E02-1039 E02-_039 12.2.2 Study correspondence 12.2.3 Shipping records 12.2.4 Raw data 12.2.5 Approved final report (original signed copy) 12.2.6 Electronic copies of data 12.3 The following supporting records will be retained separately from the study folder in the archives according to 3M Environmental Laboratory SOPs: 12.3.1 Training records 12.3.2 Calibration records 12.3.3 Instrument maintenance logs 12.3.4 Standard operating procedures, equipment procedures, and methods t3.0 Sample Retention 13.1 A portion of the reference substances used in the study will be retained in the laboratory for a period ofnot less than 2 years after a report is issued. 13.2 Sample extracts will be retained and refrigerated for a period of not less than three months from the time of analysis. 14.0 Protocol Amendments and Deviations 14.1 Amendments and deviations to the protocol will be in the form of written amendments signed by the Study Director and the Sponsor Representative. Amendments will be considered as part of the protocol and will be attached to the final protocol. All changes to the protocol will be indicated in the final report. Any other changes will be in the form of written deviations, signed by the Study raw Director and filed with the data. 15.0 Attachments None "K ; :I 3MEnvironmeLnatabloratory ExactCopyof Original Init/al Date Page9of10 Page 217 of 225 : = : _':.i AnalysisofEndogenousRuorochemlcals 3M ENVIRONMENTAL LABORATORY E02-1039 E02-1039 Protocol Signature ='='S Mar_:E (Eliefso_n -- "__ S" 3MEnvironmentalLa_tory 0_. Date: Protocol Approval Sponsor. J ..... WilliamtC Reagen,Ph.D. 3M EnvironmentaLl aboratory "*% 3M Envlronmenl_lLaboratow Date: ExacCt opyofOriginal Page 10 of 10 Page 218 of 225 3M ENVIRONMENTALLABORATORY E02-I039 i ... . ._ Study Title Analysisof EndogenouFsluorochemicailns NormalPooledHumanSerumandPlasma PROTOCOLAMENDMENTNO. 1 ! ...,._ Amendment Date: i_J O_toU3l,e2_002 _! Performhlg Laboratory t 3M En,lironmentalLaborato_, Building2-3E-09 935 Bush Avenue St. P_ml,MN 55144-1000 _'_!_ .... " Laboratory Pro]ect Identification 3M EnvironmentalLaboratoryStudyLIMS#E02-1039 ExacCt opyofOrZg;nal i_atB Page 219 of 225 o.,_q '' :_:! , "! _;_'i5 :%,! . _J i= ..'.. :'_!!t _ _:_,_ ' i_ i .._.. !_!_I : -,,J 3M ENVIRONMENTAL LABORATORY E02-1039 Protocol #E00-1311 Amendment 1 This amendmentmodifies the following portion(s)of the protocol: 1. PROTOCOLREADSS: ECTION8.1.I: Theabsoluterecoveryof themethodwill beevaluatedseparatelym human serum and plasma and ratserum. For each matrix,the samples will be fortified attwo levels of(500ppt and 5ppb). All samples will be extracted throughthe method, and analyzed by comparisonwith externalcalibration of nonextracted standards. The non-extractedstandardcalibration curveswill be prepared in methanol, andwill consist of a n_nimum of nine (9) levels, including a methanol blank. The best appropriateregression will be used to describe this curve (for best accuracy at all levels of the standards). AMEND TO READ:The absolute recovery of the method will not be evaluated as the matrixeffects render this type of comparison limited in its usefukm, ss. REASOn: A planned change in the stady's direction. 2. PROTOCOLREADS:SECTION8.2: Calibration:Cal_ration curves will be preparedfTomextracted matrix standards, in Chinese plasma and rabbitserum. The curves will consist era minimum of nine (9) levels and a matrix blank. AMENDTO READ:Calibration: Cal_ration cu.,_,eswill be preparedfrom extra, tedmatrix standardsin Chinese plasma. The curves will comist of a minimum of nine (9) levels. Reasons fornot using one or more of these standards in the construction of a calibrationcurve are up to the discretionof theanalystand will he documented in the raw data. REASON:The samples were run against 9 extracted standards withouta blank, but not all of these standards were used to construct the curve. Additionally rabbit curves were not included in the data forthis study. 3. PROTOCOLREADS:SECTION7.1: Analysisof the test and reference substances will be conducted in the 3M Enviromncntal Laboratory. The analyses will be conducted as described by 3M EnvironmentalLaboratory'Method ET8-8-23I,"SoliPdhaseExtractiaonndAnalysiosfFluorochemicCaolmpoundsfromBiologicmaaltrices". AMENDTOREAD:Analysis of the test and referencesubstances will be conducted in the 3M Enviromnental Laboratory. The analyses will be conducted as described by 3M Environmental Laboratory Method ETS-8-231. "Solid Phase Extraction and Analysis of Fluorochemical Compounds fromBiological matrices". Selected ion monitoring will bc used to detect the prcscnc_of the targetanalytes. The masses scannedwill be documented in the raw data and in the final report. REASON:Add clarity. %, 4 PROTOCOL READ.S:SECTION8_.A.nysamplewithanareagreatetrhan1i0% ofthehighesatcceptable standarwdillneedtobediluteidnttoh,r_a"ngeofthecalibratciuornveI.fsampleasrediluteidntotherange ofthecur',d'u,rinagnalyseasndenoughsampleremainsa,post-rudnilutivoanlidatiwoinllbeperformetdo verify sample values. To perform the dilution validation, one sample will be separated into two representatsiavmeples(i.et.woI mL aliquotfsorfluisdampleosrtwoI gramamountsfortissuseamples) thendiluteudsingtwoprocedureTsh.,:firsptrocedurceonsisotfsdilutitnhgesamplewithadditional matrix prior to extraction (scra adding scra), while the second procedureconsists of diluting the extract with solvent post-extraction (methanol extract adding additional methanol solvem). If the values are not within 15%of each other additional testing will be required to determine which value is a correct representation of the sample concentration. ExacCt opyofOrigin_J 3M Environmental Laboratory Page 2 of 6 _ DSt8 Page 220 of 225 . ::::'i_ i_,_:i ....+. + i i '! _::1 ,J _i+_ ,! ._.' '_'!_ 3M ENVIRONMENTAL LABORA TORY E02-1039 Protocol #E00-1311 Amendment 1 AMEND TO RF.AD: Any sample with an area greater than 110% of the highest acceptable standard will be reported as >ULOQ. REASON: The purpose of this study is to demonstrate the presence of the target analytes in pooled human serum and plasma. It is more important to show that endogenous Icvels are detectable than to show specifically what those levels are. 5. PROTOCOl. READS: SECTION1.1 The purpose of this study is to quantify perfluorohexanoic acid (C6), perfluoroheptanoic acid (C7), pentadccafluorooctanoic acid (C8), heptadecafluorononanoic acid (C9), nonadecafluorodecanoic acid (C10), perfluorodecanoic acid (CI 1), perfluorododecanoic acid (C 12), tetrahydroperflunrooctano sulfonate (THPFOS), tetrahydroperfluorodecane sulfonate (THPFDS), and perfluorooctanesulonat (PFOS) in normal pooled human serum and plasma. This study does not have a typical teat substance that is dosed onto a specific controlled test system as per a conventional GLP study. AMEND TO READ: The purpose of this study is to quantify pexfluoroheptanoic acid (C']), pcatadecafluorooctanoic acid (C8), heptadecafluorononanoic acid (C9), nonadecafluorodecanoic acid (CI0), perfluorodecanoio acid (el 1), , perfluorododecanoic acid (CI 2), tetrahydroperfluorooctane sulfonate (THPFOS), tetrahydroperfluorodecane sulfonate (THPFDS), and perfluorooctancsulfonate (PFOS) in normal pooled human serum and plasma. This study does not have a typical test substance that is dosed onto a specific controlled test system as per a conventional GLP stud'.' IIEASON:.Thc C6 compound was eliminated from the study. 6. PROTOCOLREADS: SECTION3.1 Test Substances Test Substance Perfluo rohexanoic Acid Pentadecafluorooctanolc Acid Tetradecafluoroheptanloc Acid Heptadecafluorononanoic Acid Nonadecafluorodecanoic Acid ii PerfluoroundecanoicAcid Perfluorododecanotc Acid - 1H.2H.3H.4H-perfluorooctanesulfonate "" 1H.2H.3H,4H-perfiuorodecane sulfonate Potassium Perfluorooctanesulfonate AMRNI) TO READ: Test Substances Test Substance Tetradecafluoroheptanioc Acid PentadecafluoroocAtacnidoic 3M Environmental Laboratory Formula CsFt1COOH CIFtsCOOH Co, FtaCOOH CeFoCOOH C#toCOOH Ct0FzICOOH CuFz=COOH CaHsFnOaS CmHsF_70_S C_FtTSO3"K" Formula C_FlaCOOH CIFnCOOH ExacCt opyof Or!_!n_j Page 3 of 6 Page 221 of 225 4 i' v,'_ ,'"AI "' i i ; ' I[;t_.! [_iiJ:il'',, '! _ _J } ; 'I ....i i i":"'i., ..... ', _ "., Heptadecafluorenonanoic Acid Nonadecafluorodecanoic Acid Perfluoroundecanoic Acid Perfluomdodecanoic Acid 1H,2H,3H,4H-perfluomoctanesulfonate 1H,2H,3H,4H.perl]uorodecane sulfonate Potassium Perfluomoctanesulfonate REASON: The C6 compound was efimL,mted from the study. 3MENVIROnmENLtAABLO_rOm" E02-1039 t.:. Protocol #E00-1311 Amendment 1 CBF_TCOOH CoF_COOH C_oF2_COOH C.F=_COOH CeHsF_30_S C_oH_tTO_S C=Ft_SOzK 7. PROTOCOL READS: SECTION 4.0 Reference Substances RSuefbesretannccee , ,, Perflu_,rohexanoic Acid Formula CsFtlCOOH Traceability # TCR-047 ' Tetradecafluoro heptantcc Acid CaF_3COOH TCR-267 Pentadecafluoro octanoic Acid C1FtsCOOH TCR-617 Heptadecafluoro nonanoic Acid CaFITCOOH TCR-618 NdoencaadneocicafAlucoidro CgFI=COOH TCR-036 Source SMM* 236-1B-10 Aldrich Oakwood Products Oakwood Products OParokdwuocotds Physical Descdptlon i, Colorless Liquid Clear Crystals White Crystals White Crystals White Solid Purity ,, TBD'" Storage Conditions Frozen 99.5% Frozen 97% Ambient Temperature Ambient > 99% Temperature I = 98% Frozen Perflucroun decanoic Acid CtoFalCOOH TCR-619 Oakwood , I Products White Crystals 99% Ambient Temperature dPecearfnluooicroAdcoid 1 H,2H,3H,4H- perfsluuolfroonoactteane 1H,2H,3H,4Hpedsluuolfroondaetceane Potassium Perftuorooctane sulfonate AI_I END TO READ Ct_F=3COOH TCR-037 CaHsF_O=S TCR-343 CloHd=lTO_S TCR-627 CsF_zSO3"K* TCR-018 OParokd'wuocotsd SynQuest Labs Pace Analytical SMM* 236-1B-10 PWohwidteer White Powder 96% TBD White Crystals White Powder 94.7% 86.9% Frozen Frozen Ambient Temperature Frozen Reference Substances Reference Substance Formula Tetradecafluoro heptanioc Acid C6FI_(_OOH" Pentadecafluoro octanoic Acid CtF,_COOH Traceability # 'TCR-267 TCR-617 Source Aldrich ., Oakwood Products Physical Description Purity Clear 99.5% Crystals .... White Crystals 97% Storage Conditions Frozen . Ambient Temperature Exact Copy of 0rigin_.! _._],'_1(_ _ ,_J.J.._L.. _ D_t.(] 3M Environmental Laboratory Page 4 of 6 Page 222 of 225 i ..... > :,_; _ 7:'! i 'i ,_= i'" i t _,Ti _! _:_il _:._I _i _, ! si ...._ 3M ENVIRONMENTAL LABORATORY E02-1039 Protocol #E00-1311 Amendment 1 Pentadecafluero octanoic Acid Heptadecafluoro nonanoic Acid NdoencaadneociacflAucoimd Perfluoroun decanolc Acid Perfluorodo decancic Acid CtFlsCOOH TCR-617 CoF17COOH ml, CeFI=COOH TCR-618 TCR-036 C_oF=ICOOH TCR-619 ClIF_COOH "rCR-037 Oakwood Products Oakwood Products OPraokdwuocotsd Oakwood Products Oakwood Products White Crystals White Crystals White Solid White Crystals White Powder > 97% 99% 98% 99% 96% Ambient Temperature Ambierlt Temperature Frozen i Ambient Temperature Frozen 1H,2H,3H,4Hperfluorooctane sulfonate "' 1H,2H,3H,4H- C_sF_=O=S TCR-343 SynQuest Labs Pace While Powder While TBD Frozen Ambient perfluorodecane sulfonate CloHsFtTO_S TCR-627 Analytical Crystals 94.7% Temperature ' Potassium SMM" White Perfluorooctane sulfonate CeFITSO='K TCR-018 _36-1B-10 Powder 86.9% Frozen RE,_St)N: The C6 corn )omzd was criminated from the study. 8, PROTOCOl, READS: SECTION 6.0 Surrogate Matrix Rabbit Serum I Sigma-Aldrich TCR-686 6.4 Justification of the Surrogate Matrix. Based on preliminary testing, normal pooled rabbit serum contains 6.5 vIdeernytifliocwatioenndogeonfoSuusrrolgeavteels oMfattrhiex.tesSt aamnapllyetsesshaanldl ibsethideernetbiyfiesduitbaybltehefostruudsye nausmabseurr,rodgaatete omf aintritiixa.l preparation, test substance or surrogate matrix, sample number, replicate number (if applicable), analyst(s), and project reader. ,AMENI) TO READ: None REASON: Itwas decidednottouserabbitserum curvestogeneratedataforthisstudy. 9. PROTOCOl. READS: SECTION 8.3 Limits of Quantitation (LOQ): The lower limit of quantitation (LLOQ) will be determined for each analyte. The level determined as the LLOQ must show a recovery within 75% - 125% for the analytes and must show a response greater than twice that of the blank. Exact Copy of Or]g_j73j. _ I.I.._._o-2_ _ Data 3M Environmental Laboratory Page 5 of 6 Page 223 of 225 c_ ; , -..... :tr} <:,'! i 3M ENVIRONMENTALLABORATORY E02-1039 Protocol #E00-1311 Amendment 1 A_ _ TO READ: Limits 0f Quantitation (LOQ}: The lower limit of quantitation (LLOQ) will be determined for each analyte. The level determined as the LLOQ must show a recovery within 70% - 130% for the analytes. R_,_SON: Since this is a screening-type study it was appropriate to relax the acceptable recovery levels in order to avoid re-running sample sets. Amendment Approval WilliamK. Reagen, Sl:onsorRepresentative Date / Mar/cE. Ell@on. Study Director Date i f i %, 'i :,._] 'd 3M Environmental Laboratory ExacCt opyof Original Page 6 of 6 Page 224 of 225 ! _":T :_:i '+,t '" ..... " ; _.-,, i_';,d .' _J ,"7 i _._!:_ LJ ] i.) Protocol E02-1039 Amendment #2 Study Title Analysis of Endogenous Fluorochemicals in Normal Pooled Human Serum and Plasma PROTOCOL AMENDMENT NO. 2 Amendment Date: 6 December, 2002 Performing Laboratory 3M Environmental Technology & Safety Services 3M Environmental Laboratory 935 Bush Avenue St. Paul, MN 55106 This amendment Laboratory Project Identification ET&SS E02-1039 modifies the following portion(s) of the protocol: PAGE 3, SECTION 1.1, PROTOCOL READS: The purpose of this study is.to quantify, perfluoroheptanoicacid (C7), pentadecafluorooctanoic acid (C8), heptadecafluorononanoic acid (C9), nonadecafluorodecanoic acid (C10), perfluorodecanoic acid (C1 l), perfluorododecanoic acid (C12), tetrahydroperfluorooctane sulfonate (THPFOS), tetrahydroperfluorodecane sulfonate (THPFDS), and perfluorooctanesulfonate (PFOS) in normal pooled human serum and plasma. This study does not have a typical test substance that is dosed onto a specific controlled test system as per a conventional GLP study. AMEND TOREAD: The purpose of this study is to quantify perfluoroheptanoic acid (C7), pentadecafluorooctanoic acid (Clq), heptadecafluorononanoic acid (C9), nonadecafluorodecanoic acid (C 10), perfluorodecanoic acid (C 11), perfluorododecanoic acid (C12), and perfluorooctanesulfonate (PFOS) in normal pooled human serum and plasma. This study does not have a typical test substance that is dosed onto a specific controlled test system as per a conventional GLP study. REASON: THPFOS and THPFDS were removed from the list of analytes in the revised report because the data was intended as quantitative and only screening estimates could be obtained. In addition, THPFOS and THPFDS were removed from the list of test substances, section 3.0, page 3 and the list of reference substances, section 4.0, page 4. Amendment Approval William K. Reagen, Sponsor Representative Date Mark E. Ellefson, Study Director /'" / Date Page 225 of 225