Document X86arJZQmvnx52NKrXRLV7L5w

DETERMINA5TIOTNHEOFSTHHAEKEWAFLTAESRKSMOELTUBHIOLDITY OF PFOS WILDLIFEINTERNATIONAL LTD, PROJECTNUMBER. 454C-107 /C-5 `OECD Guideline for the TesoftChi emin calgs, 105. `Water Solubility AUTHORS: RayWmionlddL.5VNanoHoonv,en, >Ph.D. STUDY INITIATION DATE: February 10, 1999 STUDY COMPLETION DATE: May 3, 1999 AMENDED REPORT DATE: April 26, 2000 `Submitted to 3M Corporation Environmental Laboratory 935 Bush Avenue St. Paul, Minnesota 55144 Wildlife I35n98tCeomrmenrcae Dtivieonal Ltd. Eason, Myla 21601 (410) 822-8600 Page 1 of 35 AMENDED 000105 WILDLIFE INTERNATIONAL Lo. -2- PROJECT NO.: 454C-107 `GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT SPONSOR: 3M Corporation TITLE: Determinationofthe Water Solubility of PFOS by the Shake Flask Method `WILDLIFE INTERNATIONAL LTD. PROJECT NUMBER: 454C-107 3M LAB REQUEST NO: U2723 STUDY COMPLETION: May 3, 1999 "Thisstudy was conducted in compliance with Good Laboratory Practice Standards as published by the LU.aSb.orEantvoirryonPrmaecnttiacle,POroCteDcEti/oGn DAg(e9n2c)y3i2n, 4E0nvCiFroRnmPaerntt7M9o2,no1g7raAupghuNsot. 14958,9 PaanrdisO1E9C92D, PvriitnhcitphleesfoofllGowoiondg exceptions eer Sre Rerdir ne iSeprearemsmdih nt characterTihzaetitoenstwassubpsetrafnocremewdasacncootrdcihnagrtaoct3erMiSzteadndinaradccOopredraantciengwiPtrhocefudlulreGsLaPndcMoemptlhioadnsc,ea;nhdoawlelvrearw,datthae STUDYDIRECTOR: y 7/ L. VanHoven, Ph.D. Scientist `Wildlife International Ltd. rego DATE Sponsor . wilt 7/60 AMENDED 000106 WILDLIFE INTERNATIONAL Lm. 3. PROJECTNO.: 454C-107 QUALITY ASSURANCE STATEMENT "Thsstudy was examined for compliance with Good Laboratory Practice Stanaspdublaishredbdytshe US. Environmental Protection Agency in 40 CFR Part 792, 17 August 1989 and OECD Principles of Good Laboratory Practice, OCDE/GD (92) 32, Environment Monograph No. 45, Paris 1992. The dates of all inspections and audits and the dates that any findings were reported to the Study Director and Laboratory `Management were as follows: acTvITY: Test Substance Prepusion SamePepcaon DraftReport andDea Fa Report Amended Report DATECONDUCTED: February 15,1999 February 1730d18,1999 Mach 163017,1999 Nay,199 Ape 25,2000 STUDYDIRECDTAOTRE:REPORMTAENDATGO:EMENT: Febuary 15,1999 February 15,1999 February18,1999 February 19,1999 Mah 17,1999 Mah23,1999 May, 199 May3,1999 Api26,2000 Api 26,2000 Sp A (oblmar JQualityH.AsCsoulreamnacen Representative wacoo DATE AMENDED 000107 WILDLIFE INTERNATIONAL Lo. "as PROJECTNO.: 454C-107 REPORT APPROVAL SPONSOR: 3M Corporation TITLE: Determinationofthe Water Solubility ofPFOS by the Shake Flask Method WILDLIFE INTERNATIONAL LTD. PROJECT NUMBER: 434C-107 3M LAB REQUEST NO: U2723 STUDY DIRECTOR: )/ PhD. Scientist MANAGEMENT: Al (Fe Willrd B. Nixon, PhD. Manager, Analytical Chemistry H-26-00 DATE -- DATE Jot AMENDED 000108 WILDLIFE INTERNATIONAL Lo. ge PROJECT NO.: 454C-107 TAOFBCOL NTEE NTS Title/Cover Page................ arrrsas ere emmsressassssssassssasssse | GoodLaboratoryPracticeCompliance Statement. - srrrstes serene essere essences 8 Definitive Test Procedure........ esas Results a --------------------_---- enn ------------ 9 SS S-- | m---- RT Conclusions. semen: ensessassnsees - -------- } TABLES Tabl1e. TypicalLC/MSOperationalPArameters.......... Table 2. ReagentBlanksAnalyzed ConcurrentlyDuring Sample Tabl3e. Solubilityof PFOSin NANOPURC WRLe.......vv mmm 2 ARYSis............ov 14 15 000109 WILDLIFE INTERNATIONAL Lo. -6- PROJECT NO.: 454C-107 TABLE OF CONTENTS ~Continued- FIGURES Figure 1 - Analyticalmethodflow chartfortheanalysisof PFOS in NANOpure Water I Figure 2 Atypicalcalibration curvefor PFO... e 1T Figure 3 - Arepresentativeionchromatogramof a low-level standard. 18 Figure 4 - Arepresentativeionchromatogramofa ighvlevel standard i 19 Figure5-Arepresentativeionchromatogramof areagent blanksample. mmol Figure6 - Arepresentativeionchromatogramofa Day 2solubility sample. ms 21 APPENDICES APPERiX TPO. Appendix 1-CrtifOiFAcaBtBeIYSI......ve Appendix II -PersonnelInvolvedinthe SRY... AppIV-e ReponrtAdmEaidmxent.... " RN. . pment smn 35 AMENDED 00110 WILDLIFE INTERNATIONAL Lo 7 PROJECT NO: 454C-107 SUMMARY STUDY TILE `WILDLIFE INTERNATIOLNTADL. PROTECT NUMBER: spoNSOR TESTING FACILITY: DLATOACANAD OTAFCSIOTPUOYDOYNF, TRHAEW FINAL REPORT: DMeettehromdinationofthe Water Solubilityof PFOS by the Shake Flask 454C-107 3M Corporation `Wildlife International Lid. Easton, Maryland 21601 Ea`sWitlodlni,feMIanrtyerlnaantdio2na1l6L0i1d. TEST SUBSTANCE: TEST DATES: FOS Experimental St-Faebrruatry 11, 1999 Experimental Termination - February 20, 1999 TEST SYSTEM: 'NANOpurs Water SUMMARY: The overall mean solubility concentration of PFOS in NANOpure water was 519mgai/L (SD =48.3; CV = 9.31% N=6). AMENDED oco111 WILDLIFE INTERNATIONAL Lo. "5. PROJECTNO.: 454C-107 INTRODUCTION `This study was conducted by Wildlife International Ltd. for 3M Corporation at theWildlife Intemational Lid analytical chemistry facility in Easton, Maryland. Atestwas conducted todetertmheiwanteer solubiliotfy PFOS (perfluorooctane sulfonic acid, potassium salt). The experimental portionofthis study was conducted between February 11, 1999 and February 20, 1999. The original raw data generated by Wildlife International Ltd. and a copyofthe final report arefiledunderProject Number 454C-107 in archives located on the Wildlife International Lid. sit. PURPOSE "The purposeofthis study was to determinethe water solubilityofPFOSat20 + 0.5C by the shake flask `method. Determofiwatnerasoltubi iliotybnythe shfla askmk ethe od is applicable to test substances with water solubilities qua toorexceeding 0.01 gram/Liter (g/L). EXPERIMENTAL DESIGN The watersolubiolfPiFtOySwasdeteatartem mperiatunreoef2d0+ 0.5C. Aprelimitensatwrays. performed to estimate the water solubility. The test consisted of additions of increasingly large amounts of 'NANOpure water to a known weightoftest substance at room temperature. The definitive test consisted of `equilibrationofanexcessamountoftestsubstancewith NANOpure water at 30C followedby equilibratiaotn 20C andanalyzingsubsamples byhighperfloiquridcmhra oman togc rapehywithmass spectrometricdetection (LCMS). MATERIALS AND METHODS `Thisstudywas conducted accorditnogthe proceduresoutlinedintheprotocol, "DetermionftahteWiaotenr Solubilityof PFOS by the Shake Flask Method." The protocol was based on procedures outlined in the OECD GuidelineforTestingofChemicals, 105: WaterSolubility(1). Thisstudymeets datarequirements unde this guideline andunder US. EPA Product Properties TestGuidelines, OPPTS, Serics:830.7840, WaterSolubility: 000112 WILDLIFE INTERNATIONAL Lo. -9- PROJECT NO.: 454C-107 ColumnElutionMethod; ShaleFlaskMethod (2)forshakeflask methodaonldToSgCyATide40oftheFederal (Code of Regulations, Part 79, Section 1840: WaterSolubility(3). Test Substance "The test substance was received from 3M Corporation on October 29, 1998 and was assigned Wildlife International Ltd. identificationnumber 4675uponreceipt. The test substance, awhit powder,vasidentifiaed: FC-95, Lot Number: 217, Expiration Date: 2008 and a reported purity of 98.9%. The test substance was reanalyzedbythe Sponsor and the CertificateofAnalysis dated March 9, 2000 indiacpiatytoefd 90.49%. The test substance was stored under ambient conditions. IntemalStandard "The intemal standard was received from 3M Corporation on July 2, 1998 and was assigned Wildife Intemational Lid. identification number 4526 upon receipt. The internal standard, @ granular material, was identified as 1H, 1H, 2H, 2H Perluorooctane Sulfonic Acid, ChemicalAbstractNumber: 27619-97-2. The standard was stored under ambieat conditions Reagents All solveats uscd nthe solubility analyses wereofACS reagent grade. Nanopure water (equivalent to ASTM Type II Designation D1193-91) was used (4) `PreliminaryTestProcedure | A preliminarytestwas conductteod estimate the water solubility of PROS. Thetest was performeadt roomtemperabytauddrineg 10.0mg ofthetestsubstanceto. 50-mLvolumetricflask.Increasingvolumesof 'NANOpure waterwere added (10.0,25.0and 50.0mL)totheflask.Aftercachaddithetconitaoinenrwa,s Shaken vigorouslyforapproximately 10 minutes and visuallyexaminedforundisstesotslubvstaencde.Afethe addition of S0-mLofNANOpure water no particulateswereobserved. DefiTnesitPtroicedvuree: Approximately 0.500 grams ofthetestsubstanwcase weighed into eaofcthrhee labelled16-ounceFrench `square bottles, NANOpure water, 250 mL equilibrated at 30C fo approximately 30 minutes, was added to AMENDED 000113 WILDLIFE INTERNATIONAL Lto. "10- PROJECT NO.: 454C-107 eachbottle. The bottles were sealed with screwcaps and wrapped with parafilm. The bottles were agitated ina `shaker-wabtatehr (Lindberg/Blue, Model No.SWB1122A)maintained at 30 + 1.0C. Afteroneday, onebottle `was removed and placed in a second constant temperature bath (Lindberg/Blue, Model No. SWB1122A, with agitation switched off) maintained at 20 + 0.5C for 24 hours with occasional shaking. Triplicate 10-mLsubsamples were removed from the bottle, centrifuged at approximately 3000 rpm for approximately 15 minutes in acentrifugeregulated at 20C. Fiftymicroliters from each subsamplewerevolumetremroviedcaandldillutyed 10500mLwith a solutionof 50% methanol and 50% NANOwpateurcr onte ain ing 0.05% v/v formic acid and 0.100 mg/Lofthe internal standard. Thedilutions were placed into autosampler vials for analysis by reverse- `phasehighperformanceliquidchromatography (HPLC) using a Hewlett-PackardModel 1100 High Performance. Liquid Chromatogrwaipthh a Perkin-Elmer API 100LC Mass Spec equt ipper dwio tham Perke in-t Elmee rTur rbo TonSpray ion source. Theremainingtwo bottles were treated in asimilar manner following initial equilibration `perofi2oadnds3 days at 30.0 10C. InstrumentalparametersfortheanaloyfsPFiOsSare summarizeidn Table 1 and a method flowchart is provided in Figure 1. Calibration Curve: Calibration standards of PFOS, ranging in concentration from 9.15 to 45.7 pg ai/L or 9.15 to 68.6 pug a.i./L, were analyzed with each sample set. Each calibration standard contained 0.100 mg/Lofthe internalstandard. Alinear regressioncquationwasgeneratedusingthepeakarearesp(raotionsofsPFeOSsto theinternal standard)versus the respective concentrationsofthe calibration standards. A typicalcalibrationcurve is presented in Figur2e. The concentration ofPFOSinthe samples wasdetebysrubsm titi utinngtheepedak area. response ratios intotheapplicablelincarregression equation. Representative ion chromatofolgowranadhmigsh calibration standards are presented in Figures 3 and 4, respectively. ReagentBlanks Reagent blanks were analyzed to assess the presenceofpotential interferences. No chromatographic interferences were observed in the blanks (prepared from a solutionof 50% methanol and 50% NANOpure water containing 0.05% v/v formic acid and 0.100 mg/Lofthe intemal standard) at or above the limit of quantitation of 9.15 pig a.i/L (Table 2). A representative ion chromatogramof a reagent blank is presented in Figure S. AMENDED 000114 WILDLIFE INTERNATIONAL Lo ne PROJECTNO.: 454C-107 Example Calculations Sample number 454C-107-2A, nominal concentration of2000 mg/L in NANOpure water. Initial Volume: 0.0500 mL Final Volume: 500 mL, Dilution Factor: 10,000 PFOSPeakArca: 687581 Internal Standard Peak Area: 603048 Peak Area Ratio: 1.140 Calibration curve equation Slope: 0020 Intercept: 0.068 Curves weighted: (11x) Calculated concentration: 539 mgai/L. Note: manual calculations may differ. RESULTS `Water Solubility "Triplicatesubsampleswereremovedfrom the appropriate bottlesafterone, twoandthreedaofyshaksing inawaterbathmaintaiante3d0 + 1.0C and folloneodawyoifan20g+ 0.5Cequilibrationperiod(Table 3). Analysisofaqueoussubsamplesafte oncdayhad amean analytical resultof 459 mg aL (SD = 8.9C6V=, 1.95%). Forsubsamples collectedaftertwoandthreedays,themeanconcentrationswere537mg ../L (SD = 27.6, CVo= 5.14%) and 501 mg ai/L (SD = 64.2, CV = 12.8%), respectively. A representative ion chromatogram ofaDay 2 solubilitysampleis presentedinFigur6e. CONCLUSIONS The Day 2 and Day 3 mean solubility concentrations were within 1o5fe%ach otherandwere averaged to obtain the overall mean solubility concentration. The overall mean solubility concentration of PFOS in NANOpurewater was 519mgai/L (SD = 48.3; CV = 9.31%; N =6). AMENDED 000115 WILDLIFE INTERNATIONAL Lo. 12. PROJECT NO.: 454C-107 REFERENCES 1. Organisation for Economic Cooperation and Development. 1995. Guideline for Testing of Chemicals, 105: WaterSolubiliy. 2 Product Properties Test Guidelines. 1996. OPPTS 30.7840. WaterSolubility: Column Elution Method; ShakeFlaskMethod. 3. TSCAofthTeFei deratlCodleofeRegudlati0 ons. 1994. Part 796,Secti18o4n0;WaterSolubility. 4 American D1193-91, Society ASTM for Testing and Materials. 199]. Standard Section II Water and Environmental Technology, Specification for Vol. 11.01:45-47. Reagent Water. 000116 WILDLIFE INTERNATIONAL Lo. 13. PROJECTNO: 454C-107 INSTRUMENT: ANALYTICAL COLUMN: OVEN TEMPERATURE: STOP TIME: FLOW RATE: MOBILE PHASE: INJECTION VOLUME: PFOSRETENTION TIME: INTERNAL STANDARD RETENTION TIME: PFOS MONITORED MASS: INTERNAL STANDARD MONITORED MASS: Table | `Typical LC/MS Operational Parameters `Hewlett-Packard Model 1100 High Performance Liquid Chromalograph with a Perkin-Elmer API 100LC Mass Spectrometer equipped with a Perkin-Elmer Turbo lonSpray ion source. Operated in selective ion `monitoring mode (SIM). KeystoneBetasil Cys co(1l00mumxm2mmn,3 umparticle size) 30C 10 minutes 0220 ml/minute: 72% Methanol: 28% NANOpure Water containing 0.1% Formic: Acid 500uL Approximately 7.07 minutes Approximately 4.85 minutes | 498.6 amu 4267 amu 000117 WILDLIFE INTERNATIONAL Lm. -14- PROJECT NO.: 454C-107 Table 2 Reagent Blanks Analyzed Concurrently During Sample Analysis _-- Number (454C-107-) Blank ta Sample Type Reagent Blank Measured a Concentration of (wgaiS/LL)Y <LOQ Blank te Reagent Blank <LOQ Blank tr. Reagent Blank <LOQ Standardanalyzed (9.13 pga)ao TThelimitofquantit(LaOtQ)ioofn 9.15 uga.i./L wasbasedupontheproduct ofthelowestcalibration standard analyzed (9.15 pg a.i./L) and the dilution factorofthe respective blank sample (1.00). AMENDED 000118 WILDLIFE INTERNATIONAL Lo. StH PROJECT NO.: 454C-107 SNaummpbleer ~~ Dayof @S4C107) Test 1A 1 B 1 1c 1 Table3 Solubility of PFOS in NANOpure Water Concentrationof PFOS Fortified Measured (gL) (mgai/l) 2000 464 2000 465 2000 "9 Mean sD ov Mean = 459 SD=896 CV=195% 2 2 2000 3) 2 2000 2 2 2000 539 Mean = 537 SD=276 508 cv=514% 563 3A 3 2000 562 3B 3 2000 a4 3c 3 2000 507 Totals: Mean(Da2aynd3s) 319 SD= 483 cv= 931% N= 6 Note: Results and comections were generaled using MacQuan version 83.3 calculations. Values have been rounded for reporting purposes. Mean = 501 SD= 642 ov=128% software and manual AMENDED 000119 WILDLIFE INTERNATIONAL Lo. -16- PROJECT NO.: 454C-107 METHODPFOOUSTILNINNEANFOOPRUTRHEEPWRAOTCEESRSING OF Prepare calibration s0t.a0n5da%rdvsivinforsmoilcuatciiodn oafnd500%.10memtgh/aLnoofltahnedi5n0te%maNlAsNtOanpduarrde water containing using volumetric flasks and gas-tight syringes. | Prepare solubility samp`laensd abdydwienigg2h5i0ngm0L.o50f0Ngarnaompsuirnetotwharteeer1t6o-coaucnhc.e French square bottles 1 at apApfrtoexrimtahteealpypr3o,p0r0i0atrepemqufiolriabrpaptriooxnipmeartieoldys,1c5enmtiinfuutgees itrniaplciecantteri1f0ug-emrLegsuulbastaemdptloe2s0C, Dilute subsamples to05.0005%mLvivwiftohramiscolauctiidoannodf05.010%mmge/thLaonfotlheanidnt5e0rn%alNsAtaNnOdaprud.re water containing 1 Transfer samples and standards to autosampler vials for analysis by LC/MS. Figure 1. Analytical method flow chart for the analysisofPFOS in Nanopure water. 000120 WILDLIFE INTERNATIONAL Lo. "17. PROJECT NO: 454C-107 lw 120 100 x ow 3foe % 040 ow] 00 00 20 00 "0 00 Concentration (ug &.i/L) Figure2. AWteyipgihctaeldc(a1l1i)br.ation curv for PFOS. Slope = 0.020; Intercept = 0.068; R= 0.990, AMENDED 000121 WILDLIFE INTERNATIONAL Lo. -18- PROJECTNO.: 454C-107 0 169 intnsty: 8955cps o 8 7 5 5 i | kh 8 |2 IK : S{0LL25 O3Lm iSIos i6si% 8715 eelis | STemne : Figure3. Arepresentativeionchromatogram of&low-level 9.15ig 8/1)standard. AMENDED 000122 WILDLIFE INTERNATIONAL Lo. "192 PROJECT NO.: 454C-107 0 189 ntensty: 32211 cps a |8 | |.o 5 " | ur = |[Rt Er I0r e25191Sm BSre 4651% t7e's 2ai1s | gTamne || Figured. Arepresentativeionchromatogramofahigh-level (45.7 pga.i/L)standard. AMENDED 000123 WILDLIFE INTERNATIONAL Lo. 20. PROJECTNO: 454C-107 AT rr Toor { 30 sa] !a i ] 70 wy !" I 150 i | i i Seito I || To Bs fh He VR Wr Wh Em Figure. aArrroewpirnedsiecnattateistvehieonrectchartoimonattiomgreaomfoPfFOaSr.eagentblanksample (454C-107-Blank-ta).s The 000124 s WILDDLLIFE INTERNAr TIONAL Lm ePRe OJECTNO.: 454C-107 21. To 168 intensity:42163cps | 8 7 [ | 5 1s. | | IE |a | to, 124 i S1T% 2Tet% eSnm zstes a61% e7T5 z8i8ts | TSiamme Figure. `Arccporonsccateatniovtfei2ro0an0ct0hmrigo/omLan)t.ogramof aDay 2 solubility sample,454C-107-2A(sominal 000125 WILDLIFE INTERNATIONAL Lo "2. APPENDIIX Protocol PROJECT NO: 454C-107 000126 WILDLIFE INTERNATIONAL Lro. n. PROJECTNO.: 454C-107 1 PROTOCOL DETERMINABTYITOHNOEFSHTAHKEEWFALTAESRKSMOELUTBHIOLDITYOF FOS 0BCDGln"fWoretreSToelsitbiingtyofComics, 105 SMLab RequNeo s027t23 Suaitatto Em3MmCoagerLsicny StPa7l.M0.ixa3n331$5133 8 WILDLIFE INTERNATIONAL LTo. Basini, Mo a22n3d60201601 Decxmber?, 1958 -- PROTOCNOOL: ASUIZGISUASOTHOSUBAS! MLAB REQUESTNO,U2723 000127 WILDLIFE INTERNATIONAL Lm. 24 PROJECTNO.: 454C-107 WILDLIFE INTERNATIONAL uo. 2 DETBYETHROEFSTHMHAEKEIWFALTANESRKSMAOELTUHBTOIDLI.ITYOOF FFNOS seonson: 3EMxvCboopcrzssicclnLibri SrCPitMmesos $5133 , SPONSORSREPRESNTATIVE MsSesABsecah TEGRACUITY 598iote mrmeiDriocvLd Bastion,Miri 21601 SDYDRECTOR: Se erin eSaSyca siirnhdichoc lan,tTh. | LABORATORY MANAGEMENT; MaWnagierBo.lfNrolxi,PsdAD. .| FORLABORAUSTEOONRLYY PropoDts: BuSimDnem 2012 0/49 ETxepmaiiomsainclaDux _3li0lt9 jose: FHCAOV pur Blkololt - | ToaSubssmebo: __AGTS __ Radwpuc --aholte i TE Wedd(2 ll 2 y(n AVE aok lle PROTOCOLNO: 4SU/1200SRA30THOSUBASH SLABREQUESTNO, 2723 000128 WILDLIFE INTERNATIONAL Lo. -25- PROJECT NO.: 454C-107 WILDLIFE INTERNATIONAL tm. 3. Wife Inerstonsl Lid.wil eIxNpeTrRieOaDtUalClTyIdOeNrmisthe waterslablty of PFOS L(ad.esnalcycail csohelmiiostcryaccfiadc,ipltoyaisniEnataso)n.,MTahrsleyn.wTilhlobsctoynwdile bao tpheecWorimed fnoalltoiwioangathlo uincepresentedinthoOECDGuideline for Testi of heicals,105: WaterSolubly (1)to shake akmctbodology. Thi dyisinendodtometdatarien oderthis rile od `derEPAProduct PropertiesTestGuidelies,OPPTS$30.7040,WerSub:ColumnElton Method;Shake FlaskMethod 2) sdTSCA Tile40oft PderalCodeofRegatics, Put796, `Secon 140:WirSoli ).Th methodofsmear quitpoviddbyteSpor be vod Wile ermsiosl Ld.Ri det forallweekporn tWIkIeItrnstional Lid ad coyofte final reportwil befdbypriestmeri aciveslocatedontheWilde IntemationalLid. ite oatanlentive cation tobespecified nto filreport. PURPOSE Th poss fisshyto dtrminthe wate siltyo PROS 20 05Cby esha. askmethod. Deteeminaion of warsail byth sh flaskmethodis applicable 1 test ita with wacesolubleeal oorcoming001 gramLic (31).Thembodelomgayy `otboepplicsbl tosta activeorvoll st bans. . "Thwatersolbilityof PFOSwilbo determined ecpentrof20.4 05C. Apein est wil bepecocmedto tims thswar kilt a1c pers,Th elistsconseists ofadditionsoficressigly ergomortof wat 0 a knowswightof istsubstan tl cobain elfociod th oobi eststo bsstan 0.01 1.Fosubstanwictesssa tinted waesoil gratertn 0.01pL, 8df test willb pec.Th dfs cist 30C, lolfoewqeudbayteioqnuof atcsie2so0nCo.utof st bt with wat a clove pert, jt !J MATERANIDMAETLHOSDS "Tht tanswilboth Bighestpty valableThe stsubstance hoiboa ast 9%. pubywreghot.TheSponsorwillbask provide thefollowin formation concsing thetest ibstfaanvaleibl,e "PROTOCOLNO: 4SH/120298/830.TUSUBASH MLABREQUESTNO, U2723 fas i : - * 000129 WILDLIFE INTERNATIONAL Lo. -26- PROJECTNO.: 454C-107 WILDLIFE INTERNATIONAL ro. 4 Comical LotorBuch Number BenT BE Cm Ethems faa ogTwhieshMtaatdfmSfIDyEDNeTtIFSIhCcA,TwIiOlNObeFwTdESTSprUovBiSdeTANrCmEBaYn SePsOsNaSyOfRo(rAtphpeandi, sia ofthstssan.TheSpon rs 4p esdest tnenlc st banecetsine thecadofhe dy. Bess "Wate thtmetsASTMTypo standards(ASTMD 1193-51)willbed (4).Orsles nybed milcid peepreionof ckslo nd lyinsada,All solventswillbeACSearngad ober, cdwillbederi tobefoeofoes tr ith te ustiattootmnisotfa. PAoprelliTmoistPootescstwaillbecondita catittesoil of he et bstanc a oom emperor.Appeosimatly 10mgof eetbtswillb lcd plat be.locrasing `kamenofwae illbe dedsteps 5 hivblr: || Tovkmeovimadd@l) 01 05 1 2 10 Jum----e----y a 2 0s 1 AB echaft ofwate vos ce,th aio willbocppodadsila Violfoappcsitely 10mites vial {est mbt ox solvedshedionof 10 cominfor soi ts tac. th ofwai, ctso thocoinswil be itarm ot reosoe a rtrdafda roifdsvi olet o,f 100mml. Fotlloy wingcippwmdd intowersly<0.1 pl. Premier tioofoscil ves conctrtions(<0. 1ttmywibth pscrmmbycmoaftistngbihetcsa,t3s,1ofmhgs.A1r0tmsessdevtel 1.cvedelshioseicngl iden cotofopel sy,reost of st bsewithpropane vosofwate maybowed. FROTOCOLINO:454120099330 T0SUBASH MLABREQUESTNO.Uz723 : . - 000130 WILDLIFE INTERNATIONAL Lo. 27. PROJECT NO.: 454C-107 WILDLIFE INTERNATIONAL Lm. -s- Di Tos Pt rcndae Thoquit of st besa satrithdesiredoan:ofwaei timidfon iraengper.eriemseirnrasytn.T(ohcvcoule)fofvwtioeeswtehdewgiulsldtpyeofteasthbsintyintromdid1 cnidcslbwielteyr. sity ibetrot 0 iof stvesselslsc dorter ableconse tri,Thededvolsofwar illbe ddd eachveelsdteves sid.Tecod edileagitoGies3i0 Lz0Ciewdkrb)e.Af |day,cos tsves iberemovedsdentedfr24or 2040.3 withxslsinoir. The Cottofthevesselwil be becentrifugedin. etorept 20C.Thomsttcnof it tac onotoaq mo vss rsd sillyfling of il pcspha wllodio. quinnpodsof2 nd 3dey Theremin 30.4 L0C. Thce toestnwilvlsebseseeelsatgcees twithiiin1c5v%o.nIcfperensi ovfatthitoesnt blbcarnvcodisnaediofuhsrphaassteooffttwheo `Spon testmayb eltwith gerquanpots. Maat Thsmt . thdfequanof htbtn willbbe procespid bythSpc.Thometedwillbe pendedtotsp inAppiIedelrod - m peat.Theo meetshbodmaat ysbocvonece roet1isopnrs.tMofohooproelidminiarynswli)l dbepyenbsdmbyith dSpa c sd StyDirect,sddomrawi dts. ColTalhteiccoosncnrationofthe stlta ccveelwllboexes i gLmg Lin - i tepepe.Thevr slsillyfortetveel withsobswii15% of chor | Eben rst `HSap"eThenSpodrsoiwdilSlantdaeynyaapci alin salty prcaios obwdwith5 hs : efacestmnbod, Allcalrctionswihsecobinedsagewillbt. - PROTOCOLNO: ASUIZEZSUASOTHAOSUBASS SMLABREQUESTNO. U2723 000131 WILDLIFE INTERNATIONAL Lo. "28. PROJECTNO.: 454C-107 `WILDLIFE INTERNATIONAL LTD. "6: SomUpeoondcTomepslSeutbisontof Rsteintgi,pnrionsofthe istmbt wd a etof hissaywil bs bpaennedowfililnbeorredtewohhfSrpc,:st nd oceans. Anyuprid nof st. 'RECORDSTOBEMAINTAINED mnRtoc:ondstobemaintainedfo datageneratedbyWifeInternationalLud.wilinclude,butnotbe. 1. 2 MAicpmyiocfsthoss guodprc oto. hm bts,iprovit dedby ist Spon: 3.. DDtaotooaxkitpionrsodsimtpcsletmiooafthilco.t. 5. 6 STtotomoicsoodcioiwnseloofphts sbstaoce. 71,. Ctonbtcennnmcnonsiiacdsoescdtnsrcooflttonrtspo,renpppsac. 5Sisaloligos. 40. T1.eAtcoonpsy.foasfmtpoen. Afn poet theesi ofEhIeNAdLyRwEilPlbOoRpTeprdby Wii rion Lt.Th p1ntNielslosk,bstootfobcttpotch mowitneg,whley.pcb: 23. DAsastoswoifccehpsttusdywgidbeythmeSdicoodmypDeeteedr. isingsycsptns toGood i 4. LabPrsac tSamty. Papo pot ss th spove ro,cai ol mendesex devises 5. TohotttrbossMinin, hinoc codemambo,pity,capil mis, oorerbm ch a,mb, thdof yl, nd ye donc roid bytheSponsa. Ducrpimof et eth ero tte etded hong with rymodicaions made. PROTOCOLINO:45412029830 THOSUBASH MLABREQUESTNO,U3 i ! . |i : . . 000132 WILDLIFE INTERNATIONAL Lo. 29. PROJECT NO.: 454C-107 WILDLIFE INTERNATIONAL Lo. "2. 7. TThhoeimndeiavnisdeenncocaantdraarondsevoifaticonhssaofmpslecalydemons a cach eral fo cach 5. persed. Desipionof nyprbienscprcoced dbo they wr sl 10. AtsittseonrtpaedpeadbfiyntihnegQrueypaAidsheeSUidsDiincg henddiMcissatghetmseosdty. peti sd i Plesnodchinges 10hpro`tCwilHlbAfONhFo fPGoRcOITofONwrCiOGteLnmodesied byeSd Do icoush prhotSop.Aony.tAenccihesatwiilllbnechonsfidoemreodf5ipdretnodf hesvricnsfleedwditwhitlhlesoaedctl. Nlchestohproto willbo dicted thf pat. : Thi staywillbocom0e0tDcLoAnBtOiRnAfTotOhRGYoPoRdALaCbTrIeCeEySPrisdeedi OECD LSBiA9t2.cm8o4nx-bdyE1PAW2(o4k30CF6tR.r7Paat9i19)2)L.BdcQhuasliatyyAcsosnatniUbysWfiodcoltisrcis nviath cGooopdiLeabsorwitGPoroadLca,bSrsatnedyPOcpeniwngilPhrporecpesfootlospporetsiioinosdopf rhes.lyAcioakimsboytf VPiradts erpioeneLpio.TehetSpboynsotoewiblrbesep,sitRlaefdoerfcoormplllawcosrkwipthrGtodoLcuWmiebddye Vinlii aalnLiossdcLoip.ysohfsrtepttrivllbcoladnbytpbreoscei t itno iilvepoctodonte "PROTOCNOOL. 4SU12029WE30TAOSUBASH MLABREQUESTNO.Uz723 000133 WILDLIFE INTERNATIONAL Lm. -30- PROJECTNO.; 454C-107 WILDLIFE INTERNATIONAL tro. . ER C1 Oirgasnis,atio1n05forWEactoernoSmoicuCloapernionaadDevelopment. 1995,Griceli fx Tsingof 2EPnroodunctMPertohpoedrtSiekseTestaGskuMeicdheod. 1996.OPPTS8307840.WaterSob: Column 3 TSSClAaTyile 0oftheFederalCodeofRegulation. 9%,Pt756,Socian 1840.Water + AmericanSRodeetyfoTesting ndMatEerialso. 1991.STaendrdVSLpeTLcfofrRoenngrion Wes. - PROTOCOLNO: ASHIZGISUASATHAOSUBASS SMLABREQUESTNO, Uz723 000134 VWAILLODULIPFEEIINNTTEERRNNAATTIIOONNAALLLLToD., rms mss PCROOJECLTNSO.O: S454AC-107 "31. We ILDLIFE INTERNATIONRAe LBuo. ee e etme --_-- A `NTIOFFCESTSAUBST TANCIEBYSOP-- ONNSOR ---- _ Tots ombiySpceer TototeMenyootbooedin hereper PEQSPclsaneSliAidPots Sat Rete Sanda ppb:Anil Sodertk NA ra Stender:_LL22HHSL Bclriane Sf Acid ! TostSut SupeCodecBuchNebr:_Lat2l! To SobtrocePy Aci logroticty:_J89__ ExgaicnDes: 2008 IL TesSubstaoceComaciizmion CedE it TEnd coRmopoA to , x HL TestSubsStotreaConndictioens Pleas intethrecommendsags onions Wiirion Lid. betes TpEe Ya__No X_ Ot pa ablynoo mics HT x MERE. - __DpfmimeERee CV. Tyltoomaion: Muomian: RaLDS02SLmgke MoweDSO NA . Adu: Iver nicky(RLCS0) Fi Taicy(LCS0) DuicimssZnel. ReTons s:Umo elln tr Tory omDanuGaltivngif0atnnggsmloferme sndPbaboecadcMiem:orSmal J -- PROTOCOLNO: 4SUIZ029W330THAUSUBASH 3MLABREQUESTNO, 12723 000135 WILDLIFE INTERNATIONAL Lo. 32. PROJECT NO.: 454C-107 WILDLIFE INTERNATIONAL LTD. "10 APPENDIX! ANALYTICAL METHOD. Aosta!Method for heDermisation of PFOS i Wer atteec)tiiPosnF(oOLtSEhMemSca)oy.obSeeaedmteprteletreimpoirnnerepdaairsgatwiooafnfrionristrneugfeoinrgttiahflipcearrteifsoopnomnsasneuc.epMieastwiidlxhbmrlvoalskvtseowdgiitalipboyenwid(lshiigslsihenpseaecmceedtm.osmTithrec TcoolnldoiwtiinognspraeysebtesscphparnogxeimtsotpercohvridoemcsoondgirteiopniscmcoendeitpiotnsnfiozrdLfCo SeqeuaantitstaidoonrocfhrPoFwOaSt.opCalprhoimcssipoegrrtaiphoinc otprOS Cromuen:tKeyHsetwolenteBtePtaackcarCd1M8,e1l00m09m0xA2H0PmLaC,3 Mobil Phase: Mebanol:ArosaAcetateBufferedWate65.35 (v=)isocratic. FCloolwumRantTe:e2m5p0eurnsi:u4t0eC aojncStioonnesV:oTlumoe: l1c0S0pry MassSpectPrekoinnEeleeAPrII:OO . RumTis: 60mimes PROTOCOLNO:4S4120290330.TSUBASA SMLABREQUEST NO. 12723 000136 WILDLIFE INTERNATIONAL Lo. PROJECT NO: 454C-107 CertifofiAcnaalytsies FC-95, Lot 217 arr Mar9,c20h00 ome `Thissamplewasanalyzed LCa AMS, "H-bNhMoR, E-wNMsR,Perndoesletmoenptoaltainoalnyses techniTqheureesuslt.s -- [ om C se -- xT --e e mo s w w o ] x [--em7 s--o uaex w r| (--emso ----XL-- b%---- CT emsox 1 eodsw --emso ek R oso enw emo 1 otin 1] moo --Tam Cmemso1 ossw 1] ddionally,Geisomerdistributionofthesuplews determlandusing. "F-NMR teckniquesaodfound ort the llowingmolepervectcomposition: hd BKe r reA (intervalmonometiy! beanct 171% Ce og Gir to. =] AMENDED 000137 WILDLIFE INTERNATIONAL Lm. "34. PROJECT NO.: 454C-107 APPENDIX IIT Personnel Involved in the Study `The following key Wildlife Intemational Ltd. personnel were involved in the conduct or management of this study: 1. WillarBd.Nixon, Ph.D., Manager, Analytical Chemistry 2. Raymond VanHoven, Ph.D, Scientist 3. NormaJn.Glassbrook, M.S., Senior Chemist 4. Keri Leach, B.S,, Technician 000138 WIWI ILLDDLLIN IFFEE T INTERE NATIR ONALN Lo. ~ A~ T~ ION FPRROOJWEA GCITNO.:L e 454C-1. 07 -35- APPENDIX IV Report Amendment 1 Original Report: Poges 1-4 and 6 `Amendment: "The pages wereschnandgesdtodiunclutdheetahelatmoenndoefdtrheepoerptodratse,erenvidseidnpa4ge nAuPmbpeErsR,GaVAn1d. Reason: To reflect the issuingofan amended report. 2. Original Report: Page 9 Amendment: Reason: nInfiornmagtiohneprreoavniadleydsibsydatthee aSpnotnhsorpurretfyle,cwtiansg atdhederdomtaotlhyesTiseofsthSuebstteastncseusbescttanicoen, Tao reflect the current test substance information provided by the Sponsor. 3. Original Report: Entire report Amendment: AalsldteetsetrsmuibnsetdanbcyectohneceSnptornastoironisn waerreeacnhalaynsgiesodtftohre teesttfshueblpsutraeintcyeco(fFtthCe-t9e5s,tsLuobtst2a1n7c)e.. Test conceiatons orginally were based on the ported purity of 985% The certioffiacnaalytsies dated March 9, 2000 indicated apurityof 90.49%. Thereforael,l daCotnacseuncihatiaosntse,stasnudbssttaatinscteiccaolncveanltureast,iwoensr,d rienccallucduilnagtendoamnindalrceopnocreinetdraastmiognsL,/mLeasbuarseedd on the 90.49% purity. Reason: Toreporttheresultsofthetestbasedon thetest substancepurityof90.49%atthe `request of the Sponsor. AMENDMENT SIGNATURES: Study Dirtorastioven, PRD. Malnagder,B.SANnailxayotni,lcal ClhseHmistory bATEA200 DATE Hw REVIEWED BY: Quality Assurance x AMENDED 000139