Document aZYNzZRzj51wg7zNDe1nzz0R

Report E01-1245 PFOS: A Reproduction Study with the Nothern Bobwhite ARZ6-185:6 ANALYTSICtAuL PdHyASETiREtPlOeRT FOR PFOS: A REPRODUCTION STUDY WH THE NORTHERN BOBWHITE FIDFaRAtSaubdRiveisqiouniE,rSeecmtieonn7t1-4 (OECD Guideline 206 LisAa Au. tStheovernson Analytical PJahnuaarsy e31,I2n0i02tiation Date Analytical Phase Completion Date April 18, 2003 PerfoarmEnviirnongmenLtsaLabboortroay tory B5u05nugn2A3v6e.0n0e SL Pack MN 55105 3M EnPvirroonjmeenctatl LIabdoeranttoriyfSitcuad#tyiEo0n1-1245 `Wildife Intemational Ltd: 454-108 Total Number of Pages 26 NEnoviparrotnmoefnhtiasl rLeapboorrtatsorhyo.ulbd reproduced, except in ful, without witen approval ofthe 3M 3M Environmental Laboratory Page 1 6n0uit Report E01-1245 open ons PFOS: A Reproduction Study with the Nothern Bobwhite PFS: A rechcton Studywin na Nota Baile ~ GLP COS MtPLIeANeCm E nSTATEa MENT ss------ Study Tie: ABnoablwyhtiictael Phase Report for PFOS: A Reproduction Study wit The Northern Study Identification Number: EO1-1245 GTroiosdanLaalbyotricaatlorpyhaPsraectwiacse (coGnLdPu)cStteadnidnacrodmspwliitahntcheeweixlcheTpoixoincsSuibssetdanbecleosw:Control Act (TSCA) Exceptions to GLP compliance: + aAsuthoemaotreigdindaaltaracwoldacttai.on systems are ot validated. The hardcopy printout"s are considered + + NTohteaslltadbalttaygoefntehrearteefewreenrceerseucbosrtdaendcdeisreactnldy osrampprlomepstlhya,sonont abeteenddaetnedrmdianteedd. on dato of ent L(isiaeA_ SAtevenSshoon,sgPiroinncipal Analyical investigator ; hh `Susan A. Beach, Sponsor Aepreseriaie 04/Dleee/os 2/03 am Emre assy 3M Environmental Laboratory Pagezazze Page 2. Gaon ReportE01. 1245 sgn - PFOS: A Reproduction Study withthe Nothern Bobwhite HORS -------------- nn QUALITY ASSURANCE STATEMENT Sty Te: A[natyisceal Phaso Ropor for PFOS: A production Sudy with Tha Noir Stay entation umber: E011245 TC(hGoAoLma)n.ial2nytinacialcraptgheasdoeinnwatshaTuodlitoewdigbyahbe.SMThEenirnognnmgesnwtalrLsabsoerpaotnoerdysQsuamiatychAsysurance Uonnt [S---- "_ [rro-- [[oCommeme |[oefoeen | w]ee] HonilytBos.rareAs HFoeuennhallef= 0918)03 Bate Stony Bret rast Page ross Report E01-1245 Aopen E0125 PFOS: A Reproduction Study with the Nothern Bobwhite PROS:A Reproucion toywih heNoho Bot TAOFBCONLTENETS (GLP COMPIGNGE SIBBMENt..rrsn Ql ASSUTANGESHBBMEN....crsssssd FE -------- SHIU OMGNON vss RElerence SUBSIANGOS sm---- SPECIMENRECEP! ANY SIOTAGE...c.corrsssnnnd A---------- LE ---- SMPCONe8C1dHAONBNISIS......cvcrrssssss 10 Analytical Results....... anar-------------- LE -------- sami SUCTOfICOONCINSIO!N crs2 Statistical Methods... m-- mmm Statement of Data QR... A Listof Attachments... OE ---------- 2 SIGANIO PAGE. 8 `AAtnatlaycshimseonftFAu:oErxotcrhacatmiiocnaalnCdoAmnpaolyutnicdasl MfeotmhBoido,logEiTcSa-l8M2B3I1f.c1e,s""Solid Phase Extraction and 28 ANChTENBt:Data SUMMAIy Tables... nd Atiachment C: Sample Chromatograms, Calibration Information, and Instrument Informton.......62 ANCHE!D: SamplePreparSaNEtELiSon ss nn185 AttachmentE: StudyProtocol,ProtocolAMendments, and DEvations.......ner 187 J aM Environment Laboratory 3M EnvironmentalLaboratory Pogo 401226 Paged GROSE7 Report E01-1245 Report 01-1245 PFOS: A Reproduction Study with the Nother Bobwhite FOS: A ReproduSctutdiywointheNortherBobi Li OFTs ABLTES Tabl1e. Summary Table, CF SO CONCENiInSTerAa ANNiGLoIVNE ...cucevrerenT Table 2. Summary Table, C4F1,SO5' Concentration in Sera and Liver with Outliers Removed ......8. Table 3.Reference SDSIANCeS................. srsem-- 8 Tabl4e. SPEExtraction Summary... ----" ns Tabl5e. SAMPIE ANIYSIS cc ssss -- Table 6. Serum QCSUMMATY TabIE uses 13 Table 7.SerumMatrix Spike SummaryTable smmm------------5 Tabl8e.Liver QC SUMMA TAbIE......c.cvmvmmsmsmmsttsnes 13 Table5. LiverMatrix SpikeSUMMATaTblye mms 13 Table 10. CFy;SO5 Data Summary, Female Quail Sora (Adultand OHSPANG)........c.rrs16 `Table 11.CyF;7SO5Data Summary, Male QuailSera (Adult and OHSPIING) uss 17 Table 12. C4F17SO5 Data Summary,Female QuelLiver (Adult and OHSPANG) .........vvr 20 Table13. C4F7SO5 Data Summary,Male QuailLiver (Adult andOffspring). ni 22 Table 14.Summary Table,CyF7SOy CONCeNtration inSera aNd Liver... e28 Table 15.Summary Table, Cyf+SOy Concentration in Sera andLiverwith Outliers Removed ....25 IMEnironmantalLaboratory 3M Environmental Laboratory Pagesol226 Page 5 Choong Report EO1-1245 Rope Eon-i245 PFOS: A Reproduction Study with the Nothern Bobwhite PROS:A Reproduction Studywith haNorthamBobatite STUDY INFORMATION StudyDirector Sponsor SWiedaintPe. IGnatlelrnaagthieorna, Lid. 3M Environmental Technology and Safety Services 3M Environmental Laboratory 8598 Commerce Drive Buiding 23609 Easton, Maryland 21601 SStu.sPaanulA,.MBNeac5h5,10S6ponsor Representative AnaiyicaChemistry Laboratory 3M Environmental Technology and Safety Services (ET&SS) 3M Environmental Laboratory (3M Lab) B9u3i5ldBiunsgh2-A3v5e-n0u9e SLits.aPaSutle,vMenNson5,51P0ri6ncipal Analytical Investigator Study Personnel `JSoeaannnP.B. GBalelaavgehresr,, WWiilid IInntteermaattiioonnaallLLTTDD,,, LSatbuodryatDoirryecMtaornagement WSiulsiaanmA.K.BReeaacgh,enS,p3onMsEonrvRierpornemseennttaaltLiavbeoratory Management HLaisraolAd. OS.teJvoehnnssoonn,,PrIi,ncPirpianlciAnaallAyntailcyatlicIanvleIsniviegsattiogrator `KOegnntjeR.nkLainKdrsutprloamn,inS,eAnniaolrytRiecsaelaCrhcehmiCshtemist Marlene Heying, Analytical Chemnst Experimenta Dates ``SSaammppllee AAnnaallyyssiissICnofmiaptlieotni:on3:1 J1a5nAuparriy22000022 archives ALlalboorriagtionarlyraacwcodradtian,gptroot4o0colC,FaRndPaarntal7y6t2i.calThreepoarntalhyativcealbreeefneraernccheivsetdanadtatrhder3eMserEvnevisraomnpmleenst,alas wEenlvlir2o5ntmheentsaalmpLalbeosrpaetrotrayinaicncgotroditnhge taona4l0ytCicFalRpPhaartse79o2f.this study, are archived at the 3M M EnvtcomensLoony 3M Environmental Laboratory. Page 601225 Page 6 Choe? Report -- EO1-1245 et A-- PFOS: A Reproduction Study with the Nothern Bobwhite SA UMa MARY i Ersoy eal yJoAr-- OAorR B e SE oni,Tosr ee res ryie s 0 alesmonmt Mtx nwaicsedordBeE0cShaSrrtssnt C A T T Tn o , etThoeeSSer , Se aSi, Table 1. Summary Table, CsF;;SOs" Concentration in Sera and Liver as P---- (me | =] -- 2 free | = | mg | FemaleControl n <t0on(1g6 m1L7) | ony| zon [a ee.|| men mastiazr " wom| 0 Fore 1| o[BmSom,m| ]| MaleOffspring. || wm | omoon| }|--w-- m ||; eens | | [mm| 1mmmv | 0 | wom || + |1 | wv| + | oory| womeas womens nen 40-1: ; Bi rL E ETomEe T m ER--e -- TALe e LSTrIntopsneng : ro. o m ar,A, pe EE WEES Seria 3MEnvironmental Laboratory nr Page 7 Gay Report e 01-1245 ee ee PFOS: A Reproduction Study etwith the Nothern Bobwhite Table 2. Summary Table, CF17SOs" Concentration in Sera and Liver with Outliers Removed =mre =+ T[e eoon| cov--omic[na|=| e ] C[ EemT emis TeTT TT TT] ] = m=mEE =mmo r eee ER SEER Ter rmare PURPOSE The purpose of this analytical phase of the study is to quantify CoF 17SOy in the liver and sera of REFERENCE SUBSTANCES Table 3. Reference Substances [Sooo ouiiens | corona rou Expration Date T 08312006. 1201/2010 =Tr Lo 3: i Physi Desrpton |VieCrainPoe| wiaesas | 3M Environmental Laboratory Page 8 GRouIL Report E01-1245 Report Eor1205 PFOS: A Reproduction Study with the Nothern Bobwhite FOS: ARprcShyweihtthoohnm Boule `SPECIMEN RECEIPT AND STORAGE NupoornthetrheBocsonwchliuosisoenruomf atnhed livnef.sampphlaesseweofretohbetasitnueddyfo(1m45t4h-e10s8a)criccoendoufcttheedteaslt aWniimkailes tflaeobcroeariatetiodornyaattlraScLhkdi.nEgna(vnLidIoMsnSem)netnnt1uoamlb3eMLraEb(noEvr0ia1tr-oo1rn2ym4e5no,tnaElO013L-0a1b/3o40r21a,.toar0ny0d/f2oE5rO/1a0-n1a1,l3y6sa5ins,.dre0sT9ph2ee7ct/is0vp1ee;lcy)ia,mseasninsgdnwewederroae Stored at 20C +1. 10C uni extraction and analysi. METHOD SUMMARY T3hMe Ednevtietromnimneatnitoanl oLfaCbyorFa:t,oSrOy3mceotnhcoendtrEaTtSi-o8n-i231t.h1e,iv"eSroladndPshearsaewEexstrpaecrifoonrmaenddacAcnoarlydsiinsg toof Fluorochemical Compounds rom Biological Matrices" T17h2e0sreatriao.saTmhpleesIvwererseampprlaepasrewderbey pmreeapsaurreidngbyamveoalsuumreinogf saewrouimghalndofdivleutringanwdithdiwluaitnegr winiha aluttoed10se8ra1o10r r1a.i0moLpofotrhe0hohmoomoggeennaitzeinogf.ivFeirvaendmiw.aosf ascheatkoneintrfiolre2w0asminaudtdese.d tToh1e.s0emsLamofpitohse epSarosmssLetdhoetfnhmrceoeuignnhatniaculgceowdnadsftoirpoa1ns0asdemidsnolutithdersop;uhgathhseethseeuxptcreoamlcauttimaonnntt(oSwPeaElsu)tedceotclhauenmnta.endaliUytp.oon40Tdhymeli.nsgaofmtphwleaetcseorlwauenmrdne analyzed for CF 50; using HPLC.ESMSMS. PREPARATORY METHOD + EITrSo-m8:B2i0o1l.o1gic"aSlolmiadtrPicheass.e" ETxhtirsacmteitohnodanGdesAcnrablayssitsheofextFrlauctoirooncohfemtiarcgaelt aConmapeosnefnrtosm bwieorleo.giGcallumedacweish,Ksauncdhiyaoshiser(acoamnmdervceira,lluysibnagllseodd) pwhaatesre e1x/2r0c,iiboenfo(rSePE)e.xtraScetrioan.samLpileesr Shoammopgleensizweedr.e iAt1e.d0 m11L10a,iuapoptrooxfihmaetedliyt1egdosferIaveorrahnodmo9gmenoafteKaonfdvyoehriwwaasteerx,traancdiedthbeyn 2Su6p0argatSanmt wofasacreatnonsiedrer,edshtaokiangclTeoan2t0bm,inleust,edanwditcha4n0trumgiinofg Kfoarnd1i0yomhiinuwiaetse.r, Tahned tanearleydzetdhrobuygh ahgcohndiptieornoerdmaCn1c8e.(SPlEi)uicdolumCnh.iomAantaolgyraasphwye-realelctuotsepdraoyf tthaancdoelummn,maasnds Speckometry (HPLC-ESMSMS). Table 4. SPE Extraction Summary [ [1 2MeWaosrusreeout(v2o0umcrovoefrsevra 0or wwoiivgwhaptreiron of ver [3 iRxisoortawOamtioorfordhomsogedrizhe woerwartsearomp ganed39n3SOzT eAdCK Ska ames Scoring. Shakelo 2mites,conto or 10mines | 5 |Ditosuporwimh8a0tmoafwratter |e |bFEortoorosragomtpnoancalyotesnroam 1mh ncoenawSihP20EmLCmleondo(oVehOHo) m._____|_ aM Emcomens Labora 3M Environmental Laboratory Pansat Page CHOU Report romans E01-1245 rAcessc se PFOS: A Reproduction rn Study with the Nothern Bobwhite AScAoMrPnLaEr COoLLrECsTaIlOeNs AvNeDr ec ANALYSIfSoto a ard raed oanrnodwaswewdonppeisrto oSe ran da of se aci, vator be lra . so or or orebapr,sanod na meraapoikoperaaandniimeaotsinc TPoeialkioh efrsoesrtavaemdewcoolotp oonaErdsr2es 0putet,erFgr.LA eer Table 5. Sample Analysis E R Ea reT ote : emit, [2] C peRp TBGT E E w "Analysis of day 1serasamples ---- SL 31January2002 [wee rie | rovrao nanao | "Anaolfycosntirols, 10ppm andoffspfrroimdnagy Ba aTT]| Are? group(day 1liversamples),seeAttachment Dfor | [Trm S raa s r | ba s| E vF a | AttaDfcorshampmleseprneppted Se EoreS eSh oieA E e FCie E faa Ea `Preparationofmethodblanks andqcsamplesal5 13 February2002 senso 3M Environmental Laboratory Rr Page 10 GROCDI Report EoL-1245 J PROS: A Reproduction Study withthe others Bobi 708Aseo n nc t ER;EaEa rr [|=[[Formboiomtumbe rao| ow oemmmm]e| apes te raged ANALYTICAL METHOD F40Saintmaacaird E TciatHeePLrSusaEadIae SVa,r vneiCltosBSSe tkceonnctoeonclure oawm7o8riSSs3s8St00m00b88Le.d2 eh seo euesan pa ranceions TaiFpdestiamtlourgethoatg,AyEhatnoocptinePnH aargr.t B 0WTEioTdoopraomkecgeomeeostmofsa8 enoeatawanmeon. 5 ou. ot Voom ee oe s oT= s T e w w|1] [oo wo Tw1] + TP oo as. sotos uese tr te styt veass3lWbireomaaesitymsUeUcmrain,heTahCeeFsS3tee0m, yy T aR EAFar. Se FEO bon was maintainedat apressureof approximately 3.0x10" mBar. EvironmeSnRtSLTaborators -- Page Cano Report E01-1245 Reponcoracs PFOS: A Reproduction Study with the Nothern Bobwhite: PROS: ARapratSiycwiionneNosh Sabu ANALYTICAL RESULTS mDeatt,a equxacleiptyt o2bsjecntoitveeds oauntdindeidsciunstsheed3bMelEonwvi(rsoanemeAnttaalchLmaebnortatAo,ryETmSe-t8h.o2d31a.n1d, p"rSoatiodcolPhwearsee. ExdSrtacutdiyonPraontodcoAln.alPyrsoitsocooflFAumoernodcmheamnitcsa,laCnodmDpeoviuantdiosnsf)rom Biological Matrices: and Atachment + SReagmrpelsesdiaotan.s.EaQcuhadcraaltlircaicounrceurfv,ewheaidghatceodef1fi,ciewntasof dapeptleiremdinfaoticoanliobra2ft0i.on98s5t.andards and Calusiibnrgamtairoixn-Sexttarnadcatreddss.taInndsatrrdusmernatngringeinsconwpcaensotcraantlibosrnatienedthdeailnypirisoorlvteontsaemxtprlaecatnfarloymsis 0.2010 1000 ngCyFSOymL methano, corresponding 0& concentrationof 201 10 40150 SCnoognr/came-Lnetxortfartsaiecorenadoofsrta3an.n9dy9aart.dos1aT9nh9de53faocnugcr/utgreaeicnveyIr.oefrTa-ehxceicaalclelivebelrdawtSiatoasnncdsuvararvudeastabandra.alcykAzeentdyicnJaogonvsweiisdoteuesdiodfon7in6e%1102m5e%etof7n0o%m-in1a3l0w%aosf dneoamcitnailv.ateIdnwsiohme(hceaesxecsep(tnieonloowfesttecloonwceasntrcauirovne cpouns, wphoiic,hankeheodUeGdh obuuttswidaesheno3t0u%sendomfiondaelterreqmuiinreemtehneti,iwtaosf iqnucalntuidaetdiownh.enTchoenmsirtucoifnugatnhieicaailobnrawtaosntchuerne, hteesnpooxnpsooaifnmtaoinitehxeicruarcvteemdesettainndgarhdes Gus0i%ngnLomOinMaSl AreSquiinrsetmreunmte.ntaDtiuon1,0thh ecqaulacdurlaattiecd Cootncbeenctarlactuiolnatoefdhuisgihnegrltehveelqusatdarnadtaircdesquwaatisono.leTnhnedaectceerpmtiendatu,ppnelr cion8cesnitnrlaetioonsruancgoeulfodr seatcxucrduayrcaisceaydmprsltieaesndrwarawdssh.deenAtleblraamcnikanyceaidlccbauyllatsthueedihtisoghhieensttocaragrliainbrarelaptciooornntcesedtraainrrdoaatwridoinmhsee(pthiokngacitchoeeph7ie5ed%m-a1l2t5:% Calbration rangeof the instrument, + hSyosotnedmoSfuietaacbihliatnyaySiacmapllersun., SPyrsitoerm tshueitcaabllbyrawiaosn dceumrovensatnrdataetderphrei0asohtresasmtpalret,antdeat fiedteonvioenl munoextprraecctiseidoncawtarsatmiaorniosrtaenddaafrdtshewebregoinannianlygzaedn,d enTdhaofptohaekruanresaepparreactielsyi.on Tahned Wpaeaskqauraels p0reocrislieosnswtahsaneq2u.a5l%1RSoDr,smseetihnang5h.e0%saRtSeDd,q1uhalpyreocbijseicotnivofesthoe stehnotwitohnamteha instrumentation was providing suiabl resus hroughout each analytical sequence. + GaCanoianalttyiiznoeundinoofgvtahCrayltitaborrngattsicoaonmnpclVaeenrstiafotircoantlsie.osns Ot(roClCymVod)ar.liatrbAraimcniksdei-trlueemdveebnltyamplaalscsiritx,ngcwaCiltbOhraVtiowInemricehooefpcok2riw5ea%ds `Samples bracketed by one or more failing CCV's were reanalyzed. Limit of Quantitation (LOQ). The lower limit of quantitation (LLOQ) was defined in liver (Twnhige/hg)uptahpneedr7i0nI-ms1ie3r0uo%fm na(ounmagi/nnmiaLfl)atcarosintetr(hieUaLlaoOnwGde)swtiwahacscaenpdteaafnbialnleeydteexapt5reaahckteeadrcecoaanlciaebrneattariasottnioc2nu(romvfeestphoteihnethitbghlhaatenskit.s. + Balcacnekps.iaMeectuhrovde bploanmkss(epdurtcohcaasiedbrKaatndtihyeorhaiscproinnskienogfwtaeterI,nsstreuameannt.dlor vr taken through tChoentaemniinraetisoanmple preparation and analysis process) provided a measure of background + QouxatlaicttyodCoanntdroalna(lGyCz)adaannddMcaotrnisxsSopdikoefst.hrOoonevosest ooff nainnaelyQteCisarmppiclaetse.in eAadcdhitmoantarilx.wwaos mReasuilssarpekassalmopwess:ai the lowest concantaton were prepared wih each extracion set Eonar nosy 3M Environmental Laboratory Fuga r2crzze Page 12 coon Report EOL-1245 Reponon12es PFOS: A Reproduction Study with the Nother Bobwhite PFS: A Reproduction ty ith th Noth Scat Table 6. Serum QC Summary Table D F Reem T Iipa----------[--a ----) MSSppbt | 2otngmL | 752% 30% som | song Gestppm | To10000 ng aw QCeoppm | szs0ocongml | 1a sre | Saarsansancopniceatarbaebolrs5TMheSse043S0aiwaorewparropnearcvaalnloownitnygpaataeredp1roOcCe.drsan fc daay no GfCrestaimee wih anand 0 cor rl eu. Daa hom (anys wt ced Hla Table 7. Serum Matrix Spike Summary Table EET `ConcentrinaDtiitoend J {Three Matix Spikes [7 Thee Matix Spikes 10000ngm | 93.9% 1 --- I --or--e--tso el He aaSols PART Ir ---- oor Tabl8e. Liver GC Summary Table ioeteJOoe Dinlutged sLPieP Soren soars T [inR o F[a } wwe Te mT aGCssampalesweraeared1 crs ga es.Ova fom ana wae a eiodod in conaning Table 9. Liver Matrix Spike Summary Table ET vb J------ Four Matix Spikes FouDrupMlaitrciaxtSepsikes 499 ng 110% 4990e0t9 i} 2% - Tmuhsetprboetowcoilnan7d5m%et-h1o2d5s%tactreittehraiat.2A3loftraelleQoCf tahnedvaermi"nQiCm-u1m0pofpb%sopfieQsCeaxceeaecdhedletvheel Stated criteria therefor th Iver data reported nthe ow ange of the curve (Day 0 samples) aM Ervonmens Lavoro 3M Environmental Laboratory Page 1301226 Page 13 Cone Report 01-1245 RoperE01-1245 PFOS: A Reproduction Study with the Nothern Bobwhite PFOS:AReproduction tidy with heNorthamBobi msiagnyifbiceanbtliyasaebdovheight.he lSoiwncreanagllesotfutdhye scaurmvpel,etshethoavterwaelrleimnpoatct"D(aoryb0i"ass)hoonwdaatnaaliystemilneivmealls or none at al. + A`cbosmoplaurtineg Rtehceovreesruiletss.of tThheeexatrbascotleudtequraelctoyvecroyntroofl tshpeikeGsCquasnatmiptlateesd ivserdseutseramninmeadixb-y etxhtarnacwtheedncuursvienganadmaatsrolivxeenxttrcaucrvtee.d cTuhrvee.serTahsepilkoews qsupainkeisiaatreeh3ig7h.e8r%ushiinghgear,sotlhveenmtecduirvuem. squpainkteistat3i1o.n7o%f htihgehetrh,reaenldevtelhse ohfigshpiskepsikeussin2g6.a6%solhviegnhterc.urvTehever3s2u.s5%a maavlerrxa-geexirhaicgtheedr cisurovbesedrevmeodnsftorraItveesrtshaamtpileiss.appTrhoperilaitveer1s0piukseesmqauriaxn-toaxtteraactte4d.s8t%anldoawredr,s.6.T4h%e shiagmheer,traenndd 8av.e4r%agheigohfer3.r0e%spheicgihveerlyusfionrgtahosollovw,entmecdurivuemvaenrsdushiagnh esxptirkaecsteodf ftihveerlmiavelrrimxatcruirxve,witahgaainn demonstrating tha ts appropriate 1o use matrx-extracted standards. + IPnFtDerAnawlaSstannotdaursde:d fTohrequianntteirtnaatliosnt,abnudtarwdas(PuFsDeAd)towamsoniatdodredfortograollsssaimnpsltreusmeanntdasituarned.ards. InusstirngumtehentconRceesnptroantsioenCoaflitbhreatCioFn.SH;PLaCni-oEnSoMfSp/eMrSluoirnosotrcutmaennetsruelfsopnoantse,e wpoatsascsailubmrastaeldt u(sPeFdOSt)otphartepwaarse asdtjoucsktesdolfuotriopnusr.ty aAnndytfhuetuproetacsaslicuulmatiioonncsonrterqiubiurtiinogn ctoontcheentmraastisonosf P10FObSe expressed as a percentage of dietary n-lake must adjust the dosed amounts as well DATA SUMMARY pTraebsleenst1e0d tinhrAotutgahch1m3esnutmCm.arTihzeetianbdlievsidduiaslpslaaympClyeFdSatOa. Rceopnrceensternattaitoinvsefcohrrionmdiavtiodugarlaimnsjeactrieons. cAlallidbartaatiaornecucrovrer.ectAeldrfeosrulptusrtreyloaftethoe nPFolOtyheresfaemrpelnceessteusbtsetd.ance used for the mairix-exiracied IM Environmental Laboratory 3M Environmental Laboratory Pago 141228 Page 14 Chau) son2 Report E01-1245 PFOS: A Reproduction Study with the Nothern Bobwhite 08: A Battin Sit npuyn itt Table 10. CyF17SO5Data Summary, Female Quail Sera (Adult and Offspring) 7 y as CX TN | coos 510050,tomtom, 01512 | vo] -400 |. E01-1245-28446/454-108-332, Opp female,au, 01-31-02 o0o0ss ""o=o <400 [or | | ovsmalomosommimnmnoa | y E01-1245-28449 454-108-336,:Oppm female, adut, 01-31-02 | * orraeniss se 108308 oppmemain om onaroz | ; |cozroe]e10s0340 cpmtema, san cor 2 EE ------ |covrosz00s0 osmezr msamiczodron | || oass ?1 w 03 | ! To2zE | |: oor 3i o1 s i oorf 3i | <"a0 400 W oo ww =|] S"awon So a "wR oo E 2Zo aw [mesmmn normre |I | | coriscandce ca3 opm resn. Tol | = |. E01-1245-281 468|454-O1p0p8mf-e3m5al6e,, adut, 02.0102 1-1245.28470/454-1 020102 0o3s | 1 es ! <"0n | <d00 =n <d00 | cova car 64100300omtem, san 2012 a03 1[1 wens | G&L |] 2 74|a54-1 male, ack, 020102 | or | Eot12452847 45 10836, 0ppm ems,oi,020102 ] Te oncomone Eo 2852 007 oppmiemae somGov Ga EEOo0112--11422454552--220084448873544455544--111000860.-3377724,,, 1o100pppppmmiffemeaol,e,msaaddanh,,l00022e00011-,00022 02| 050024 E01-124528489)104pp5m4fo-mal1e,0a8c:03270160,2 | 01 EOT-1245-28491/ 454-108-380, 10pprmfemale,adut, 02-01-02 Tv ||1 1|| |__1| | 7| F 2050 EE <n [| ED ET 9140 aemwo T | seo em [T-- 3M Environmental Laboratory rats az0 Page 15 canny Report E01-1245 Repo cov12cs PFOS: A Reproduction Study with the Nothern Bobwhite: PROS: A RerokSteytwiohhneNorham oth ci . Py - 0) pI Bris bu poe ie 01124528450]454108.92, 10pmfom,au, 02.0102 E0o1n1122445522004106]]55411063066,61o6p3ppmfo8emmaal6e,,as,oum,00220076022 | o0r1.11224455220655020]4551100603930,, 1pppmmoonnaall,,aacct,002200110022 012185042]45441063504, 10ppmoma, ck,020102 | Eot-124526506]a54-108-506, 0ppm female, sc, 0201.02 |Eo1-12452650elas- 106-398, 1oppm female aom.c20002 | |[|"ECoort--1122445522685511154]5541.0180-6-44002,, 10ppmmfteemamle.,asck, 0022.0011.0022 [coE0r11122445228655116]75454110008-440068, rpppmmeomamasasao0a2o0,r10a2s || |corresaasele1020. romeocen cars | | corasscase sI e108250, opmtome og 2vaz | rors wm [1 wm | 01-124528524{454108273, Opp fol,fpr 2012 03 | |owe | s| | 0a ||vnoen | ||| 00s: |[ewmw [| | 0s | maw || ow |T v ae 01 | + | oso | | o0r2 | 3+ || mS0600 | oa |v OF Ve Tw OoFr "a00 acs T a oer mm mens || & ] adsesas 1 orn, 0201. 02 <0 wo wr EE ---- 01124526550] 454106.525, 10pmfom, rg, 020102 io 12452050) % vom] rm rm ne -- 10538, 10pm fama,org,0201 aTe | 1 | rToesmo |] ow [1 12300 12500 00008 T1 w12o1o0o0 | | corrass uses 454108587 pmiemal ot. oanz| 0% ! 01124520506] 45410856,10pm foal, orn,02.01.02 3 T T1500 10800 [ i* heTh"eArveeraargoetowfo on res. DruespulStsaemppa"rtceofnornthese samples. 3 Boi values ae included and averaged in i+ntTehvea,sefovratlhueesrwejeerctrieojfnectoeudtebras.sed on th resus of the Dixon's G-Test, 50% confidence Accuracy: Sample resuls are accurate fo100% +1 30%. S6ra QC samples met th acceptance criteria or the method. aM EmvoomenaLasorsory 3M Environmental Laboratory Pagn t6ct2zs Page 16 canny Report E01-1245 Repo gor 1245 PFOS: A Reproduction Study with the Nothern Bobwhite P78: AReprcton Sywith trom State Table 11. CsF/S05Data Summary, Male Quail Sera (Adult and Offspring) ho [ie -- PAraleEtar ErrOetE i A ---- | eo-- pep---- - | eontassc2o0t7-|14524.e145h058:2305 0cp4o7mmmalk,,at,230,013.13. 12 [Ee rm mi, a0. cm |cove wspms stream oad oir . 0112452045252 454.4108:-39,100-359,0ppmmmla,aht,01.3102 01124520184 AE 464108341, 0Oppmpmmaall,eeadk,01031311.02 EE ------ . cor 245220046574554108c3m4mmal,,a0,203,102131.02 BE124528150 54108.347 opm,a8,02. | Eot-12645206446611 454.140584.13048849, ppmmmall,,oodt201020102 Er ------ 0022 1s <w0o | <0 oa T[ e= - moM 3! FD 0 oz i wo | 002 | || oo | w= o=z 1 v ! a p"w oz 7 wn o1 mwn |Tw 0oz T! T e=m w 0o2o 1 I <400 [= -a T3 "waoon Taw oT Em Tw i E01-1245-254674410|843s545,-108:358, ppmmmaal, atG2401-02 | owe xTT! =8 0 c12o0:m ommemae, sa | 0070 || 1|| <400 [w- n | conto cot-12652007 454 108.3p5om5mmata,di,t,020102.102 0o2 | 1: | -0 | cot120 12652807 454108361.1, cpmmopma, i,act,act 020102 I o-z |13 ""ao aw [| oron1.216254522884475 T445410895p5pm,mp al,atp C214-- 2 oy ! "-a EE ------ Tm Tw |. 01-1245:28478|454-O1pp0mm8 ale-,ad3ut,602-701-,02 | o0r1 !: p5y | <a00 Emon Lasoo 34 Environmental Laboratory Pan 7ct2ze Page 17 Cadeue Report E01-1245 [emp PFOS: A Reproduction Study with the Nothern Bobwhite: rAoRat.on Sty inrarSmite saw fr el [ET rea O ER E ee E |co orer m opms mnsnse ozare) 3 | i | ow ER | conressamor sss00ur, pommansam arora| G3 | 0 |corsoantn ora.e sansnzm anaz| 0} wo | mm [vem asso os tosom. mansan ovomoria| ok | 10 | ewe | wom [covrosszn sa100006, ppmmat sam arvoczonaa| 02 | 1% 7 A [corm serio o mni aos r| 02 | 0 | | eo foe E B E w= Wns sors 3M Environmental Laboratory fo Page 18 caduin Report 01-1 Ce 245 f-- PFOS: A Reproduction Study with the Nothern Bobwhite pe 4 ital EE | 01-125-28519 454-p1pm0mal8e,o-fsp2ring5,023-01,-02 RAY Ln oY 1 Ld = 0a 1 40 a0 =ETE= |. E01-1245-28522 |454- Opp1mm0 ale8 ,of: fspr2ing6 ,02-601, -02 oor 3 <400 vrwnoromrmmeem--me--neTee|[|| =E2 T| 1|l|:| 21=23T|[ET]| 27 |454-108:291, x 02:01: | | [E01-1245-28529 |454-106-522, 10ppmmale, offspring, 02-01-02 oor | 10 19300 19300 = 1 = > [enmeom [orsa mrsnmemnz | oot | | | [esePmmeemem vmmeen| 2EEE|| [e-e] o L | 10400 3M Environmental Laboratory Page 19 Cada Report E01-1245 Aon E1245 PFOS: A Reproduction Study with the Nothern Bobwhite: PROS: AReposition SuchtheoremBtu Table 12. CeF1SO5 Data Summary, Female Quail Liver (Adult and Offspring) oe [R-- } G.F.50. frie Saers Duco .| [[eeovvirotaeomer o[oppppmmaatissoarriimm [[Eorioizams joppmaisssFuver [[[cEoorr-aiazsoeanamsl[ooppppmmaeirssosFoirvuemr [[croromateeoiz omemm aike Fe [[FFEEoovriroatuezacnoteusrJooppppmmaarisstirriiame || toowwe || 1 1| | wwoorr || | roe | 1| wer | || ttoosws | |1 1 || wworr || | om | 1+ |r | || toomoe | | 11| | woorr || [econr svzesousntobomssotion [`[CCoorrrieaouzo5s0s8o73r[elo0oppoopmmmaasilsSssaFraiLnrneerr [Feoriatz sot opomats60FUr ee ree] JE[Eootr-11a4o2:t3026207_0[05Joppppmmaa1so3o4FFlLivveerr [Eorioszsoon Toppmarororiver Erma = = corisiz0717 10alSp74 mLin | tow | [or | Loassizm |1 | | 13031 1 wor "or | ome[1| wor roses| | osm | 1| wer | | toms|1 | soo | tome | seo | [e[oEr01--r1a34i2z:o30r7r5s3 o1p0pmpmaarias37o8fFLiivwer toss| 1 | wo | [eE o omse [10 omT a.86 Five [E oes| m mw | E01-1342.30610 [10 ppm ai. 204 FLiver [Eor-1542.30964 [10ppm a.400 FLiver ot sez 1000 om | 1 swe | ror T Sou" [ER -- 3M Environmental Laboratory [-- Page 20 Pr Pre] Report E01-1245 Aopenots PFOS: A Reproduction Study with the Nother Bobwhite: PFS: A Rprchein Sayihth cham Baba --Su iT.-- viobusosr : Saeunt y i eor-10651104 (MG/G OF LIVER) oraz i 20 oovt11906653311221004 [00apip .22p75a0mFLm vaeerr,l.Otspraogepmg || 044% 05008 v T 200 200 [Eov-1085.31224 0ppmai27 F Ler.Oteping _|__0.4526 : 200 [coE1o-1v0r65o31e2s50a_t[1a0ipoom [1i0SS12p75F ovveerr,mOOttseappiinnrgg|| _oe70 05720 || 1 1 |__| [E01-1965312o6i06_38[FL1ve0r0, pm Ofsping|__0f120 11_ aso_|| [EEoott-itsoe6s55011227050 [1100pppmmaais1 s FLwireo7 rtOfispF prrnngg|| Oodbsot || 1 | | v sssewo || +inteTrhvaels,efovratlhueesrewjeercteiorneocftoeudtebrassed on the results of the Dixon's Q-Test, 80% confidence Accuracy: Sample resuls are accurale to 100% + 30%. {oG+ rCLitsvheaermmpQeltCehsosad(6m.)pAlwleesrievceiwdrihnGioCtn mscietatert,mhe(th3p1e)roselfpfoiGrekCeetdasahtetesa1ec0choplnbedvweOleCmruecsrt>i1ebr2ea5w%wriaesicomv7eet5riy%n-.Ath1la2lt5o2t%/h3reorfiavaleOrC were win 75% 125%. aM Ervonmens Lavoioy 3M Environmental Laboratory -- onPiage 21 Report E01-1245 pon 011265 PFOS: A Reproduction Study with the Nothern Bobwhite FOS: APeprcShctinonha NoBac m cE Table 13. CoF S05 Data Summary, Male Quail Liver (Adult and Offspring) ne ET SYS,nse Spa [RosJse Ro "ates [cot10420550 [pom ai 301 Mer [ortonfopmarssomue =: ET | toss| +| wow | [[Eorsizs00se joppmaicariver |Eor-tacoaoie Jopomaicoomiver [Eortotecoieo foppmaisermiver [[Eoriseacosss loppmaiseomive [Eortosooiseloppmaiossmiver |tooo | 1 | woz | |oes| 1 | | ross|1| sor wer | | | owo | 1| woz | | tie |1| woz | [Feor-1312505o1p0ma. 251Miner [[Eor-tasecoses Joppmaissomiver =3 | vows | 1| soz sor | Eorioizosed opomaisTmilver E01-1342:30562 0ppm a. 3M5Li0ver E0o11--11334422-:3300660106 0[p0ppemmaa.i..336613MMLLiivveerr |Eot-istec0es loppmaisssmtver | tose | 1| <2 | 1T.o00s8s4 [1w1 or <502" | toot | 1| sor | [omanTomarsain | Eot-1542:90600 [10pomai.371 MLiver [[Eorstza0m0s[roppmataramiver 1 tion Jw 1 oese | to | | vos | to | mer esr | eeoor | foe| [eor-toss0rea [1oppmaiarrmiver |Eot-1342:30[1708p0pm a.38MLi1ver [eros fumes | oz | to | eo | 1.0501 10 un || mer or.192230047[10ppm a. 367 Mer [[Eor-1st23006s[10pmatsemis ime[0| exo | too | 10 | rower | [Eor-tatesosot[1oppmasssamiver | tose | 0| eee | or19425005 [10pam JMLver |Eor-1542:50[1007p3pmai. 401 MLiver |E01:134[102po:ma3.04039M9Liv1er Leonse23tcar[roppmai orm voz Wo om 100 | nosso[too | som seo | aEmon asco 3M Environmental Laboratory Panzer Page 22 6nres Report 01-1245 Reponon-125 PFOS: A Reproduction Study with the Nothern Bobwhite: PROS: AReproductionStudy ith ho North Bobwhite assu 2 -- | poe oIu) LEC a Eyy Nah [Eor-106531164_Joppm ai 2ssM iver Oteping | 03005 [1 | 200 | [E01-126531199 [0pp a. 256 MLive,Ofprng 05047 wo| [Eor-1[03pp6m a5i.23661MLi2ver0. 0 Ofsprng 02613 i wo | oswo [ | 1 aeor | [Eor136531220 [oppmai 2saMLverOfisping | 0sess | 1 | 00 | [Eors5a1204_oppmai ze Mer Otsping | ose| 1 | woo | J[Eeoot-sTos653311225655_[1[01opopmmaalSss2a2MMLbveerr,oOfpsipningg || o0s7o3r25s || 110| | vaesrsoo || Eoor1--11296655.3311226750 [[1100pppm.mai 4552MMLLiivveerr,,OOffiispprinngg | | o0a5t3253a|1 |I ser | [Eot-136p5pm3ai1.56288ML5ive_rO[fs1pri0ng | 07501 [er| intTerhveals,efvoratlhueesrweejercetiroenojefctuetdtberass.ed on theresultof the Dixon's G-Tost, 90% confidence Atachmant 8 coneins data summary tables. Accuracy: Sample results are accurate to 100% +- 30%. f*orLitvheermQeCthosda.mpAllesvdeird GnoCt mseaemtpltehse (J3)osfpQikCeda iga1c0hpapvbewlemro u>b1o2sw5i%tthrienco7ve5ry%.- 1A2ll5t%hcerrerviear wGeCresawimtphliens7(56)%-we1r2e5w%i.lin criteria therefor the second QC crieria was met in that 23 of all QC STATEMENT OF CONCLUSION C`SyeFryayrSe0s4ultIsn 1f9roomf 1th9equmaaile(<c4on0t0ronlgigmrLo).up Tphoretisoenraoffethsitssstfuodmy sthheowfoemdalneocdoontierocliagbrlooupevpeosronf oG`1fA1ntah1liy4st1is0cta0lu0dryongusihtmosLw.feoCdr.tFnwoSendOleystescat(rarabalsnegaelge9vee4ls4s0of0fo--Gmy2t2h,e6S01O00s0pipnnmg1/d6moLos.fe1Td6ambbalilereds1g4)r(.o<udp00AsnnhagoiiywimecLad)la(rnTeasabuvlleetsr1a5fg)oe.r CSoivFe1nstSoOeyn (srearnagesa4m7pl1e0s f1r4o7m00thne 1L0.ppTadbolese1d5)f.omale group showed an average of 8670 ng/mL oSfer6abriersduslt(s<f4r00omngt/hmeLm).aeThoeffsseprriangfocsounltrsolrogrmoutphesfheomwaelde noofiscptreicntgacbolnetrolleveglrsouopf CpyorFtyi,oSnOo5f ihni6s Sfteuscuylsshfoorwfevde nsoerdaetseacmtapbllees lferveolms tohfeC1yF01p,pSmOd3oisne4domfa4lebirodfsfs(p<ri4n0g0 gnrgo/umLp,ShTaobwleed 1a4n). aAvnearlaygtiecaolf t1h2e60100npg/mmL.Go(sreandgefe6m2al2e0-of1f5s2p0ri0ngngGirmoLu.pTsaheow1e4d).anAnaalvyeircaaglereosful1t2s4f0or0fnvms.era(sraanmgpele7s6f4r0om18500 ng Table 14), bLiivdesr r(e<s50u.l2t nfrgo,m tThaeblmeal1o5)coanntdolqugarnoiuipiabslheowceondcennotrdaattiaocntsabolfeCleoFvyesSOosf CinyF1,oSfO2y0in(21099 onfig1,9 CTaabFlueiS1O4u).in T2h0eofe2r0 brresduslt(s<5f0r.o2m ntihge,fTeamballee 1C4o)n.troAlnaglrytoiucpalsrheoswuelsd ffoiro tdweelnetcytaIbvleer lseavmelpsleosf o47m00t0heng1/0gppTmabdloes1e4d). maAlnealgyrtoicuap esshuolwsedfoanwaevnetraigveerofs8a8m5pl0e0snfgrlogmCtyhFep1S0Opypm(rdaonsgeed5f0e1m0a0l-e group showsd an average of 4900 ng/g CyF1,SOy (range 3290 - 7550 ng/g, Table 14) aM Emonmans Lasser 3M Environmental Laboratory Page z2s Page 23 CRAG Report E01-1245 PFOS: A Reproduction Study with the Nothern Bobwhite Table 14. Summary Table, CsF;;SOs" Concentration in Sera and Liver E" TAT Cr CR ares Pra=y]| || - ERTS nl <acongimt (1 17) =Nofsereno|rI SAvereace wr| nr aoe eC mamma Seckebey ' Te Male | Foro | Contd Group - | I - Ee To oo | ee. eee FE r [oE o [en[+[oer] re | -- on oni Report E A 01-1245 J PFOS: A Reproduction Study with the Nothern Bobwhite Table 15. Summary Table, CsF17SO5" Concentration in Sera and Liver with Outliers Removed [[ie |mor[m|e wew FemaleControl [mmmy E=e|e=r [nn|er w -- oreo ewe oT|= RR EA T GSareamGmro||v|[ | | e[ew r[| e[mrer] S EFom Eem --E e ----E r m re r, STATISTICAL METHODS `STATEMENT OF DATA QUALITY 3M Environmental Laboratory Page 25 GRALDS Report E01-1245 ReportEO1-1245. PFOS: A Reproduction Study with the Nothern Bobwhite: PFOS: ARagrStuodywidththueNocrthatmBoobwhnite `The protocol and method state that 2/3 of all QC and a minimum of% of QC at each level must be within 75% - 125% Criteria. Al three of the iver "0C-10ppb" spikes exceeded the stated cbiiiatesreida,htighhe.refSoirnectehaelllisvteurddyatsaamrpeploerstethdatinwtehreelnoowtr"aDnagye 0o"f sthheocwuarnvael(ytDeayle0veslasmspilgenisf)icmanatylybe. above the low range of the curve, the overall impact (or bias) on data is minimal or none at all. REFERENCES 1. Wildife International, Lid. Protocol No. 454-108 *PFOS: A Reproduction Study with the. Norther Bobwhite". 2. Wildife Intemational, Ltd. Protocol No. 454/120700/QR/SUBAS4 (aM Environmental Laboratory Project Number U2723) "PFOS: A Reproduction Study with the Northern Bobwhite". 3. Centre Analytical Laboratories, Inc. Study 023-044, "Validationof Analytical Methods. F"ErxotrmacSteironumoffPorotAanaslsyisuims PUesriinlguoHrPooLcCtaEnelseucltfroonsaptreayo/rMOatshseSrpFelcutorroomcehtermyi"caanldC"oEmxptoruacntdiso.n of APnoatlaysssiisuUmsPienrgflHuPoLroCocEtlaencetsruolsfpornaatye/MoarssOtShpeerctFrloumoerotcrhyeimniQcuaalilCoSmeprouumnadnsdFLrivoemr.L"iver for 4. 3M Environmental Laboratory Standard Operating Procedure ETS-8-231.1, "Solid Phase Extraction and Analysis of Fiuorochemical Compounds form Biological Matrices". LIST OF ATTACHMENTS Attachment A: Extraction and Analytical Method + Attachment B: Data Summary Tables + Attachment C: Sample Chromatograms, Calibration Information, and Instrument Parameters. Attachment D: Sample Preparation Sheets + Attachment E: Study Protocol, Protocol Amendments, and Deviations + Attachment F: Example Calculations IM Enironmertal Laboratory 3M Environmental Laboratory Page 26ot226. Page 26 GHAGOY rmeR Report E01-1245 +705.A msat y rn n tt PFOS: A Reproduction Study with the Nothern Bobwhite `SIGNATURE PAGE We cert that this report is a true and complete representation of the data for this study: (AuhSovPonwo =oul Pons Aran vestigtor UTV baomz nmaarnaggeomoeont Em 04/18/03 sana 3M Environmental Laboratory ---- Page 27 Report E01-1245 apron 1265 PFOS: A Reproduction Study with the Nothern Bobwhite PROS: AReprScsicnhuaNcottaSoains ATTACHMENT A: EXTRACTION AND ANALYTICAL METHOD, ETS-8-231.1, "SoLID PHASE EXTRACTION AND ANALYSIS OF FLUOROCHEMICAL COMPOUNDS FROM BIOLOGICAL MATRICES" SMEnon ay 3M Environmental Laboratory ---- Page 28 Catan Report EOI-1245 Ropon Eor-1245 PFOS: A Reproduction Study with the Nothern Bobwhite PROS: A epaicton Sywi haNornScbmtts 3M Environmental Laboratory TT hed T SoldPhaseExtransdAcnaolysnisofFuorochemical `Compounds from BiologicalMatrices Method Number:ET58.231.1 Adoption Dat: 301 Revision ate: 3/5/02 Enc Doe: 21/2 Ady Yr bFiolrainn MRaargaee 34Toe8/08 Envios aay 3M Environmental Laboratory ShCoc mpsesahi an pPsil fern) Page smn Page 29 cnet Report EO1-1245 RoponEot-1265 PFOS: A Reproduction Study with the Nother Bobwhite PFOS: ARprSotudydwihuheNcorttonBoobwnhite 1_Scope and Application sTohlisdehiedudeceisot(hSePncE Tchscfdetoysyoesbsoemxefrs. 10voelt,eotsoe.gcoriee spaovdidr eudnseh, sesinng lyona sme 2 Method Summary s16 hns,folrolgerGsietr,5cdeiebdynebylhe yes.asypreip g(eg1estdredesdlsrasAvPs ort oof she HChoamCraasororinobsepsoeneps3ee04.ssaTpsokriep0ehrreapcrpstnph csrop1n0c.3cTohnesurbceel,edmcacp.wri.lAm cera,sia np(atCcKeo)e dtiseacdudce. d3 re Ceonioonned ChSuPcchotridoe.oFtraly.thoe bycosnofe oresscareacniefs HbPeLSCPEEScSoN)rndeaayged by he 3_ Definitions 3.4 Dilution rAieSshiohneprrsmods 1. fs end 1 i.ofsic + mksof 6c or lof 6mL bis, 3.2 SPE cartridge A5p"crooslgouonncso1liuneritnnevtahecp,nocsmoppeoousnrodfsoFcinedabt rrneeeceoddndSoamVcEGk Neciltohbnbodnodregd2sxi)cmcrslcreshreantdteroobodevrednrd..10A1 one sensfor abi 3.3 Reagent grade water Wot ithetal conceonft ere st). 3.4 Quality control sample Semple odomanior he orci fie (osmt ik)dvr hecota cc fe 4_ Warnings and Cautions 4.1 Health and Safety Warnings Alva sporie ove, ewer dco whenwoking vith snes, sp dorce en. Uwsieection it hevoagecbsfheprobe,Wheneng.teprobecpp.solapeofppeimaeyS000 42 Cautions TCakoemcper4odtrialelobehnesShPowc1ob 0in p nsuagfher heSmF eSaklaaddtersashes. ARhing Sod Phas ExractEiornsnadaA s i nofsi acs hs Compounds fom Bop Matrices Page2010 SMEnvtonmontl Laboratory 3M Environmental Laboratory Pagesors Page 30 Cadul Report 01-1245 arEot125 PFOS: A Reproduction Study with the Nothern Bobwhite 7705: ARapSccwuin acriroBonte DooteprLC helnepesshtn edoywo.f0or(50 eck ron, boc eso ce 40 [---------- 5 _intorforences F Ce ee e ------ 6_ instrumentation, Supplies, and Materials Tmh olow.ing npoenidn,sspecrs.y nderdisefeeed5sdb peng hs mel. Eq 6.1 IonnsterumenVtaOtRioVno Coe? i VicamT unTy2 tsnis C$15oBmaciMoian0ti 0 Ske, Each WR Mtierram01a005r) ion ie inp MowSecor qedi lpn onion ce ASRN ---------- 6.2 Supplies and Materials Drpcecnegrsepsontc pcs. pee F Vereo acklm sco,eAfo li 0r m 1 Le) Foamsc reiiiGt.EWN)homor erpeie Gomisabes, bap, 5d0. LGobt pcses thos SaitnesTocDieefrscbacofdi0e0in00S.kofshes) Sm P xinrcos ndr| oSrose kpiccnr se eCuWnn) Crimp snwiiohon Si PCeoEmpxocEomisBasoepoafoers ci Pacers SMEnon abrir Pan arcrzze 3M Environmental Laboratory Page 31 ner Report EO1-1245 ReponnE01-1245 PFOS: A Reproduction Study with the Nother Bobwhite PROS: AReproductionStuywih haNorbomBowie Cringe HPLC silcol, seis obedemi bye altoddcr nve edi. 7_Reagents and Standards Rene rade waeMIG,Nap 1.orsive Acca,HPLCgrade cam Metro, HPLC adeoreqns: Arsm cot reegetnreorcq oidfsor bss, oore sper 7.1 Reagents preparation l200eMcs cocn caocntao nsoogne.t Wediegvhs ar ikp nla0l}3i 00 sg c monliey lcbeen,0Pvoheui:tacan2g 0vm0rl aiewr. toe roomcen NnodtoeWpheenparnedprriecnttvoorceessodnndhnogeytdin eget,ortrly dnd repusn, 7.2 Target analyte standard preparation Plreopwairengryatncautrlsooynatn) ofhresymrotdpcie. riicoosnaeoseeyrs s drescs epte. Th sWeinghdsphoaknoye1s0pofr9riemsnriigtn.c mk 10 ol,solacefk nd cond hecs) weiignthe Bring volewihmenfor sock dodofpeony 100pe(gl) Dkiueitne.s1o0k00sghlexn SihAmDed.rol=fSrOHgwio.rkingsar |scksionofappei0npoam.eElxye Duibitownor:kiSngsmasnd01mwLiAhOmDOaLn- Ofogpiroc wokingsandr) 2stn ofpox $0por. Exar Dciltawno:kinSgOspgmodxl |1 w0ihneLn1d00oS0primodg.uc.e wakingsind honofspe.0.50po.Exe 7.3 Surrogate standard preparation Psaenpdespmrporsgtiosnsu. as.The loving anexe lyand ya ytbesprog for llsurge Weighposi5t0110 ofarog ed 3 10m.ienekad eon ec wight Brnocluewithncnfor og srsockofspoximtly 0-100. PlrimemeesdaaTgn kussrndsabdingdravvsloasonnrscishmi,taTnradssbwporkekosnoeyrdT|nolfTo9f0.i 1 bpeeasm naRndeedse ckaco0r1l0odne 7.4 Internal standard preparation dPepapenl.sds)The longs 2 earl ond nyor ey oeapc folmrt Weighpron 50110 mgof rlsind 0100-,vlametofksnd codhesc eight Sli PhaseExrctiEonTssndaAansysoforocherica Pagesofy Compounds rom Bitopsi Matis WM EnvoomanaLaboratory 3M Environmental Laboratory. Pages20t26 Page 32 GRA Report E01-1245 Repen E0v.1245 PFOS: A Reproduction Study with the Nothern Bobwhite. PFOS:AReproduction Saywin hoNorthamEcwe Brin omewithetn fo 2 es)stadsickofpricy0 100 Prleapamreeris fekra)dsraingdwoakiengsihdme.anTefrrsworpkiingSnadyar| olfo0f.1 0r7s.s Resd toacak1cda3c0hea isdandsndSanson sb hook or SPR a 8_ Sample Handling sis eee ene ad stbeesren ni he cctionisprone Allowsais oh rom pea rie xc. Tpryedmpoveaihlacwilaisliaslsys reitheechsd ys. Excsandsa sample are soredin aaybenewdiy ltbe mdseec, cxaicedisisd ndsos plerfeorymedberefered poise C nef 3_ Quality Control 9.1 Blanks. 811 Solvent Blank vain fens wd eek. Soeblka okxc. 84.2 Method Blank nCreqdaonfd 1y0 mid.ofvwitohoe1rploewpeh:ssfna.ai(dwo5petiddbWhksaFrogweatnhdbk1sr Shey 943 Matrix Blank Amd n8pporsfo1pi0ema. oMrrLigoafneirsepirdcofhoomorgeeof ceencde.1o3nsbokc.oOroemariFaesmrsyobye Cionlfassnd)lregrceionvergdewesirnd,+mspcuolrcvecitndd23CcCc oVmfoer3m,o ottosf ln .eTehlecuoer fccife0shatenua he1idseyspred eino.ndd(scy. nvei dforn ce, 19:1.31_ ingSumlycyowoolomtacie1 fh0e.s8ycoorlortenid sdfore crv, f(4mei barks 9.132creCopmempereotc(4h) Peotnitbaaessiecnsg) ostmseuove mi cyomvmberinc(epotci wiahorehe 3nd ond ed) 90.33sanS:ucroommermihoyvvalilee ovsooxppcirfosrd(4)rhiesbaarve, rnegpcrfooumre(4)lyiabblkewmivn of secs lysdho dt 91.34elylinrtomcarn i is vaw le,encofeed dk aksrybe aenddwll edn 9.2 Sample Replicate Samospinswepreening ch dypesoon rotvine Sli rae ExrctiEonTnsdaaAnofsForcyhersics `Compounds rom Blogs Matis Pesos SMEnvtanmentalLaborato 3M Environmental Laboratory. Pages3al226 Page 33 Report EQ1-1245 Rogen E011245 PFOS: A Reproduction Study with the Nothern Bobwhite PROS: A ReproductionStudywith ho NothamBobwhite 9.3 Surrogate standard Scioaaernocgosncdearn.icomapanrosf osftiaey,clsanecue,5widh tielx sop p ofaWnidk ppoisexdciioentde SFyuiorcoarltysraeaygbnae shsepaed0dmndseppllkm.otthteeoe110rpl 0iri.ce o Aee demowib glEhndor,mr ieereo vEefGooRcfn Osrgciieoonne,ndHSowre, 9.4 Internal standard aFiclsoacnanreismecidocmnrpgoerotafbofecilsbsyiconolunmamep.lesaepk wih rlsanse cnc oon Type memo sins iedso he 20mL.ofext ne 15mcng be,befor antiote 9.5LabControl Sample Labconesles ro comportofhsmeh, 9.6QualityControl (aC) Sample SPreiiptaaolresnubcirutnseciosonaevned.(oTQrCo)opshoalmQCpLSermovcncr oesexsopcieen efafirn.3fdstoSvoeryma c1oine0dpsurmesofte oTfgaQiheCssw(12im) qececmcctoncbteoplsreaepusHd(iQreCe)rccwiaolcylhbeeepvernoeondcaOcoChdcyh1ofsipultdeuxpvncpihoednc.oos( necoofhaoec yoydyeA miI rshin praieey (rQCesacpeswsilpclo cfooeesrplGadtcacahboefracneoefsoafoa.nThveecls isdbelow mybe edsnd Lovie X51 XbeL0 100. Mibene casas pitcreid ofbecltaioncin, [rr---- o c TuoQiCawipc nei,ilswh haeofsObCesedr oOCspiteriaclbbye ieytSahevserpBearfksonstSionngdrsehe ellets tcabyotbt hntadt COCC rpescgx conweihofpheiecytrsoyplsicmetsbeesey hesme bid rn.AnyOC moles c1xrciiocnsdeomsmtolbexecd dotc cssyinosys ntheQC slsfrombe sespl ExicshcrOCci,scaodediisnhoowfn1c2aufee OoCf7s1l35s0of opchevi.l mAmtineitmasc3r.fllIOckC,slseesosrtecocetr beardeid 9.7 Sample Dilution 6Anyaslaimpmlieownichiv.Ivcf epaiesnoAtdof hehieghntecoafbelce daunowrillseaodexithedSlkoeemnee.of3 snk alton wlbsproms veryrie bes. TSotpnsfeo tihred muotioen doron1,oeosmaarsiswilobe espSbpi1n0or lpt aintndsrmei e0y. So PhaseExacionETsnsdaAansyioflaches CompomoBougine Mdavser Page or1y SM EnveonmanaLato 3MEnvironmental Laboratory Poon a0126 Page34 Cred Report E01-1245 Roper E01-1245. PFOS: A Reproduction Study with the Nothern Bobwhite PFOS: A RerStoudydwhtuhe Ncorthter Boobrnite hs eicdoicwp ciutmwheiasdttdhsie dtoiilov.nesnntposoetstoefpxlveea)otrothn(ecnextrdhcattnidoolnes(ixunagdctatawioipnnroiacd)eo.sw.hliTlmeheetthfheiosecpoooncvdoerdraosrc)chonescisotsnosftiofndgieitnheg IdFettehremirneleawthiviecphervcelnt15d:i3fufrcncte EisecnoCknwtiioin 1o5f9t%hfesoamtphleesctnwceonsraamiplcens: addins)testingwilberegefo 10 Calibration and Standardization 10.1 InstrumentCalibration Oexwntieoccltolcnondisitssoto,cfsas3vdmeewelrli ibnedporbfu eyptanhreee(dr9)oreoyvmeaesnx.dacdAtdeoidtaotnrsicsaitlnabnondaatrkodnsc0urivheessmmaeybmeaextaactheesodnaslpea,tpeerssuapyi.e c"eTnrunisfegee1w,0biw.ionrgtabdrpaospabplerploaspcmpriuip.etof.Tdhiedwilclobnirorlepicdeodrwbhoimopgreenpiazreidngcaohnqoletssfoerth0e s81t5amld. cruermveo.vedB.esmetomixorshake fhecoolmixconsebetween ako10 re horusseple x Recordcachsnd vou:onthewighhohamessector exictionwashessppoprse. Four107ml.aliuos,or otherspeopiaeamount, ofcosrolmati serv 5 esblanks. "T`ohnedssenrdvaicnunrdndvtec.eospntAscae0emn1pil]tseiosof.nT9 shatnanmdduasmprbidkesirnfgoocfmmsotsaundiasrbdlssaaennkdds.binnTldafbkloree ye| bnineydabdeiluaatrsekeda,aewhdpereenpeaapmrperedodpnrbiaystteh,oea0naoiltykhsetescOnoCed documneanottoeied. URseeeAtdascehcmoen1nC2t.1ocroatlhuebrtaeptphroepsrcii)cfooncme,nt2rataionaodfincyleeiaancdhcelcbroontocnesnatnodfsntdeOwoCkaimnpglset.ands. oHTocwyetvoepre,tmihotevmcaianygagbetelhess1pla.i0kyeymdidsatnmapnldlaer,dnoiotishpe hmhoetii|kOptLhreo1100tdelldLh.idanmigokgweierthiovnraeodrgs,eanimipleedtrseeumoipvteeeddmfohreoomxotfvognee.xnecistziadoelnde. Aengryesaiotnenoxlcyssitaesdimng hiepessanrderacsuofrtvheeprairotseamchs)toifnegxiMtas.sTshye uobveeresquuastbiolnewsiolfltbweadreeterminebyd mAhtnesyelbe3veewllietovuhetilsnsimd0ey%7b5oe%fdoerc2itn5iav%la.ioefAdmlilinInavayelosmeessttbsoehrdoeawocsetisvsawtpieodl.sbaeedrreeesereraynnt.e-ecauolotefdt.heexbcleapkt.heALmLOaQwrhioachf Solid Phase ExtractEioTn aSnd Anofs Fuooy chems ical `ComfropmBioolougicnalMadtrisces: 3MEnvironmentalLaboratory 3MEnvironmentalLaboratory PageTof19 Pa3sg012e26 Page 35 Ghd Report EOI-1245 Reponon12s PFOS: A Reproduction Study with the Nother Bobwhite Pros: A Rptchcton Stywihth Nother Bbw EpS rovi co-- nan) -- 7 --7 -- p musk ar iie--Us| --5 in Fina 20.--veume | [s7 [ ums T 1t0i |T WGoo0mmssooo mmnml | |--Osn tonngmo] [sE m{eTSS T 0 7 t11e7 ne nT_o--TmEmrio ] S So v o o T_T 17 || [n[ irmo S50a TiOtu1m7m7iaem nnnl| | a Dloiosmp]| Samesoar| 0|emosporoant| oognt 11 Procedures 114ObiTifsseue rSamnplselePsreparation CeupeeALo e1p00 n fsqyt 0s.00o05)woneber eoepBeoe.sos set o sel. hip Weight decdpy csl e Readewiht cpetcl co wok oeppp con Hekmuaed tis bo A825Lf agen fcr eci smo oer ter sil os oleecon by he td raed in ke oomeoeggapefeesrle. soe UkTire ie nhspladgdfo apmin2maniiey, Ronee robe terwhecoi4n1if 0ng lw e hPmO .of cg grewa. eemind Ti1kmesoptioemreesnponasroIeOnnoith(ws1ii015i01e c0rcchselo fo.rid Rr c.To5r29d 43s21e0wi0h2.5eeoesofmheveiri) oost To omysgdre ni le Sod PhaCsoerasosmicsoshoomnsdraoyepscsoMf oPvoesres aEmmonsLasorsony 3M Environmental Laboratory meets Pogedsorzze coPatge a36 Report E01-1245 Repo EO1-1245. PFOS: A Reproduction Study with the Nothern Bobwhite: PROS: AReproduction StudywhtheNorth Bobwhite 11.2 Fluid Sample Preparation Obifrogen fdsimpleand ko 1thw roomemptortr SLeeobtech1e5dmwLo.rpkoslhyepertop(ylAeaneccheennAostuibewritwhokshleisecety)foarmdbeern,tsaimpnlehIeDc,ixtitnagSdc67ct.snidaonalny nas oVorrtlexsmic hepafledvsiealngolelfeorsaappprocximaielcyo13ntseeco.nds.then esr 107m oflid,ofaber appropri amon Retun sedsamplestofeefeentationmoushvbec ened Aanddd$0omcLuofmrneeannenottweaoeteidto he 10 nL.of idfor 3 1 dln,o otherdlaion a deteinedbythesly s1oamveodlluanioonabsearbtohvaen.10misremovedfor inal volun,sist the atervole:sccodiglyfo siti he 113 Tissue and Fluid Sample Extraction GAfhr teeod1hdossammnplpeosehgfvoeebsepipernoapnrieempiacrdeldyac15osndeingc1tohseetnniodnnsss1e1.11a.0dn11o2,vaotrheespioeshapkveoblyeuhmaen,d1h0e3. clean 15 mlpolypropylene ceifoge he eu misediidhomogenized portionsoefee fercxactonamounts hav benened. Resa thevolumeremovedontheextractionworkabet (AchmetAor ila sect Spikebbs.amples,andsad eyforcxicionithsop sandnd adscibod in this eto. SpliikbeatciaocnhucavibecsntnidonasdasnadneddemcahtOiCwsiatmhptlhee. propreawn ofsad ssdescribedin hismtdfo he Vortex mixthestandardcurvespl ndOCsumplesfosspeoisicly seconds. Tsoecnadcsh.spea standard. 5d 5.0 mLofaccion, 3p.3d voi mioshake byhand spproimstc1l5y 3P0la0c7esplosnnthpelmsoodnetshesshdskienr etacnosngp1r)o.prnesped or20mics toadequacy ix (3stingofsppeoimaily Rseemionvgeoffapopmeuthseimaikleyr2n00d0centoifngtehoetmoadseplroipeidtne scpeendf6o.111.0mitesto adel pelle the peecipie 4 AcSdaolnut4on0t.h0CesmoLppeoasfnadrmneistxenbdiygoretahvdeeetwitntcsiv(te0aecl0amnmesl5.0t1bnetl,hickiinpntcgaerenrdboee.ofRnsermiocovnesannpayylobefesftcrehoamangitehd.eicecln.dthisefnsgasomp4hlee maybeput lo the cninge tbe sndthenteweade. sch he esr 0thSPEcandy sch esr aridge ut103 acummail. fNlOoTwEo:f 5W.h7emninmiin.gFhleovwsamcya.vsarey rheouvagchucurmticdugnebseraantd ahepPrAaxaiycbe1SrKePAdftoor isvowanpbepsardodwyiicnsiasiilocrn mosfavebeendrdomwn. Pr(eOpanrEeatlhiequSotPsEocfrwiadteg.eabykgwicanreg nwoiteowsiltohwtshpeprcoxoiimely0r50unmL0odfrmynecnsoaf,erleachowabsyhwspeosde (wo Atfrhsttrhinecgooilsucmuonmtpolgderty,mpheosu.tthesample ilthe reservar witsndallowal ofthe quid to pass Ruth vacua onihfospproxmily mites 0adel doycoh SPEcards. Place colction15ml plyprcplen ceufge beunderschcard ndchewith 20 ml.of tinal. Spike exactedblanks, spies,adsandals ilnssand3s describedin is tod. Sol PhaseExixtiEoTnaSnd AnaoflFlyaroscheimicsal `Compaunds fromBicogical Matrices: Pagedof19 SM EnvironmentalLaboratory 3MEnvironmental Laboratory Page 3701226 Page 37 Chen Report EOI-1245 Ropon01-1245 PFOS: A Reproduction Study with the Nothern Bobwhite PROS:AReproduction Sad wihthNorhamBowie Laitbceachrdhsesetoni,I1MsSsmpaober,e,t,wiltthssvle,y Stmbeesc,ovmolpaflne(iaechdivde m.bcesi,nwiople. rc be.an ly)efibexin Tonercachchant gsaisnd ap. 114 Extract Analysis 114.1 Software set-up yOtheoMaosLy.isndp3ge,er up arpieeitwroin.gShevepeTtsi3hsanchdsr p nFor 2 hsof Example Sample Ls YYMMDDS T2200 1st ssnrn A =Ani) YY Ton@y MM Tsmonth 02) DD~ Texdoy 00) = Ftsani ife oy be caspe wildi, ee, a {Aeeinn flearmebesringseeisnisahn1enddee sta irco ers asht 2 iirsaeof,be ttyarancy, 3 36 Example cane: IVYMMODHPoAE20401 =i oframemne YY=Totyen MM Termeni DD=Tordsy t= iteiqabeersniglwilh gh9000) Anbdtsame doefsrhpersnse.n, sameh (MS) einone i. emb a ee ctionsrhe TiMoRrMc(tMmAileiei ,RebsitecoksnctMaaeinbonond)a.ErSentbdlttorpznttonsMMadSSeoS3vusemsqsPpuanrecaedhsnsodemhnsosdSI1RI0fS4iM9ngSl0eSolamReinepeoprsnp4) oaea. andrxsPosotfo magiroahnomuion aybe colle. ee iran MasLymn "Gude sTynpe telah sq eins wi pn isi vr ks, etofcd mix Sampyle dxcpsrreesayeod irhwopQoCsselsteitcedrhseracnrdyr 01scamoplreedcisnp.lSvxecsbabnksyshoubde ied sh Soi PhaCsaeEmxrpcotisonEronTmdsBaloynsgissofMaFtircoesemics a Envcnmens Laboratory 3M Environmental Laboratory Page 10010 Pa38gcraezs Page 38 or Report E01-1245 RaponE0t1245 PFOS: A Reproduction Study with the Nother Bobwhite 705: A productionStudyithNorth Bobwhts 1142 HPLC setup Setupmrp scsiotheseepeeshove. RSeectondteesHPcLCotnte:sloinncocmioncohoooosrsphpesshotsiindsppp foeps spor. [------ Inctnyic 1 Coens = 100mies Powe 00min Mobi pe:Soe A 2m ArmAccs,Sb B+Mel . Se sitpn rogr | orge [wT o o =I= 1 |T= 1 = = 1143 Instrumentsetup RFeifenrcEhToS5n:A28,mOepseFnesodMoanciroaneSoof bue.idceess Qa Tri QuinteMssSpcomtr Checkinsb velinHPLCsos nd lf ces. Cm oc essiisve c 3 1 ia s p dofct opsr,.Usneemehepxroveciheeckcecoi.STheesshcolbceufnawd.i Tamang Opn mpg. Cenper ts Hock nddse cns. Open it Eder. DoriontheHPLCrch andnesober flow begin semcquibrien Ste fovto 10-p 50 op: SaMPLCpmp "0 Ober dopeocitcing fpfere.Afoishudbe expel with ritopenkisendhe of hert.Reo iheptotpomfisbbreed otcqo freon 1mies. Toit ners pnts Dry 20-0 coor ESwhinogs 10-15 eens Sold Phase EnvctioEn snsd Asnaisof Forces Compan rom Bos Mars Enon Laser 3M Environmental Laboratory Page tof Page 3901226 cn Report E01-1245 Foponors PFOS: A Reproduction Study withtheNothern Bobwhite: POS: AReproduction StudyiheNota Bout HPLCct owme fw 10-501 rss 400 psn ts. le sree HPLC sprogcorey) Sorel mesoepimely 50C Dontacrpeanc pesimy20C Thee sigs eyhrs mci ini pr: iCtottheoemose,snpio5.anscsion dodomMas yo nd slyinde ihcopy Cflokobnsga au to i t te ActtineConor ldPiun 0ciofbes sony sy cp Masons 12DataAnalysis and Calculations 124 Calculations otredothos rFc, ey wlbedsnec rees Coke te tisamcor be tldnsing lowe ction: Argon + -- WEEVIL) Msi Emon) ae IVDV (ny Coleco cocoa ofse cabinsdssig lngcasio o SokeConcnitio pm Spied mo), 00g 6 on (gor) = MataAmount gmor mm) 3 Wcl ig ue 10 10m) IV = vole DSVV.=SDoineiscsolram: smegnsnfrail 10m AR Amfomy sys sey [LE -- FV Faole MA = Motos Sime Mhofcsndconve,sumedtoe oS mkver role Ma ssi le xmlofaml nlva) Calne piepen nisin bincin heey = E bred Reni aE s Bik Resi, Soh PCeoEmxpnacntErioosnmndEaArrioissoMfoFvlesac! Peron SW Emcomers Lavoro 3M Environmental Laboratory Pagesoi z2s Page 40 Gnd Report EOI-1245 Repo ots PFOS: A Reproduction Study with the Nother Bobwhite PROS: ARepetitionSywi aNorham Bite Culdevit nin u lovins qin Relive Sandord Devin SSNIDEMOn gp Calinperndevation ig ollovgcq: uhraion EEBSEOCEonpcCCoonoe Con, " ClbxualcormaionfyinBad pn) AR gin DF +f dssaldol)* aT lin * Las = te) Cust conentiofloen ineUs AR)DF ~dens inl) abil * 10g = Ge) snp) Fe) nbs 10003 13 Method Performance 13.1 System Sultabilly SiySsitmnesiAistepyenladlsbeedhepteosdaseserpiesiotf ee3reensidn (heprrcoilmopnlsicoanfctd,hcwsdplckeuePsryi wwtllbhhee STdo5nEecReSpeDae kheprcercyotvsg ieenecni cioodn ahg earcuhsretr05ch-gA.F<o2TheewhchhAaFns5rhe RnrS.ayo ebeeskeoanl tbes 20. 1S6aemoy paenthop somoee igbeit,Stsai ymaetrnclaricopieRdpioe,0 eE in 3m codeytloef 132TheQcuoanitintatofioncin ln oreclinsve,ptebyeres in hepkssof he i ms e orb Ahlysivdecevu op et posiibeie in25: 6fi hei oewilh cpofBeiLO opo, Crutganostdsbewil obkyaksrsobctloirsrheyce ri Bk wil Be dence gy abdcabot cae cori est cepossesacl LOG thc cotmehr hesimplest bssyedrece 133"ooAiccuorfaaclyuty cone sis nd 72h lity consichee ie oped shown pike Soka PhaCseoEmxcmtioonEossmodspoipesnoMf aFovcirscherict Pe of Ecomat ano 3M Environmental Laboratory page stot 226 Page 41 GN Report EI-1245 AoponEon 1248 PFOS: A Reproduction Study with the Nothern Bobwhite FOS: A RoptSyowidthhieNtchotmBoownie Suieypecedonsdsncoynmfaisr.ls1thhoisdeScsTo,nmdhssoyaesvecs3oopndevsorlooedcwevlieeaoanee< hd0e4vr.iraDlwvielle boe.dceivrdeei dis 0ngre windl obe 14_Pollution Prevention and Waste Management Semplevast diporedof posi bardwsconser. Fabien vase desocf ghBTUcones. Gispipe ws dsposdfinbeng consis sid ear. 15 Records Csoampttecxucionnhs cheboth thd ater pla ket,adsoeith he dy aExmcbhepragse piectnedefhoos,aly vetgcommianod.e sFrpilc iefo,rmsatcioniipdoel. dcindcoyr 1eaofNn)rc i rmnys be ed ppthc e ay. aPtine eg e aagee,srims pele oat,rxdciesri,on method omMany clk wih he cy owds. Coy Ptthecalbasionuve be pop resin. Pi bes: gapndstsor ihesly owdt. wiwct rionsay, gionmh nd routs om Mas, strwihesd aw. Sumit diai ise hare ce7 IMS)andsr nthesoy elder, Bdacoknuy cdte.or ta 0spp edn. Recon nlynto he Fi re:and cio ofbck 16_ Attachments Aen A: Excion Wokbeet cenB:Sle WeighVosWorksheet J ------ etc Di SueyWakes 17 References ETSh.a0.n24A, OnpesronPnadneMoainreSofotohe"Micros Qua Tre Quupole Miss Specter Fed E1552. Oprosnn Ms osToec Grn" 18 Affected Documents Nae Solid Pas ExacionEssndaAnisysofarches Page Mof19 Compounds rom Biologist Matic SMEnvrcrmenaLaboratory 3M Environmental Laboratory Pageazai22 Page 42 Gdn Report EO0I-1245 FoponEon12is PFOS: A Reproduction Study with the Nothern Bobwhite PFOS: AReproductionStudy wh haNorhamBotta 18 Rovisions ReNvoins7iion Mion formaning changes AddedRdelsedo esmsaotitoinntol scons soemhe ReDvuiieon 02102 exerancnio.n odor,aml proce, 4 clin. Add sashes 1d So PhaCseoEmixnscEossmnidsBAoalnoingssofrFesroscherical aM Envionmana Lasoo 3M Environmental Laboratory Page 15119 Page 301226 Page 43 ane Report EO1-1245 Reps on1265 PSSerup oNouimba: oe PFOS: A Reproduction Study with the Nothern Bobwhite PFS: RorocictonSty win NotherBoba Achmet A Exinction Woes VotaRospsione ETS8231.1 Samoa Tinga: rT -- rT77 [ 117 [TTT rT 71 1 TT TTT T 1 rT 1 rr 7 171 rr rT TT TT r L Tremmpodi ioT n ---- 1 -- T TT 1] A nnmatmatt1 hrhn m m reme one Sa pen Wa o H) E 2 tr ge vonp pany rm westSk Bo on seoscht tbasnps n toop op e sam wkcchorweonnt 1roe edelvoeveimmereaatsrehsaipaes nstA$i0r4i. ehnlse et t 7lomtmy to Sd PCeaErxotmoaentromsbeieadnf ternis etary Eniormariat asrsoy 3M Environmental Laboratory Pagesat228 Page 44 CHAS Report te E01-1245 mm PFOS: A Reproduction Study with the Nothern Bobwhite EBeEeE E-o rer rr 1 1 rrr rrr rr rT I Ey 1 [1 | I 1] | ---" rr ---- ---- -- 1 -------- 1 -- [1 Fr 1 TT I 1 1 -- em ie een GRA 2 Report EO1-1245 RoponE01. 1245 PFOS: A Reproduction Study with the Nothern Bobwhite PROS: ARoprSidoywidhhiaNtomiBootnte AnParevpedtino:: SMaemlhpoldeimraotvriiioxn: Aachen C: Calibration Standard Concenttion Worksheet SEtqaunidpamrendtsonmubmebrer: FiBnlaanlksoTlvuenct nordFlTi:dideatifir An`aNloyltteemmiisesstdd aapppprroonx.. 0$30000gwidm:: ASvuerrmiossdgsadapsptprooer.es1.0000vpups. J : SiAdnsaoivnice amiped| FAt | nAol| oATl wn ob | vonleCs:|| DFiasodn|| DTenosiety C CE -- 1Se 0O mint0| 0 gmt [CoC sCo T -- rT o --s7 o0 TO oo mar oro]| C-- a 1 O C1 YT, C1K 1 B wt1 in2 a C[Ceooo I Tov w T TiTm a umtesrieTr uoiree0r|| edWcWil Calvaconcnrions fsandarsin elatonto he el20.soeadial ti FFrale|| SSecrornees|| AFmale|| SSoveroogsnts|| AnFaaidic|| SSadreroonpee co | vom | om| ml | on | onl EE Era EPoom]sTsuw meF5s8w0Tsnwr s5w0|sewem|] to Pe me) [ior ew [aoe] om) [oCa ian]] ia] [mswoom]m me] [[Ciia]] Solid Phase ExtacioEn anTd ASnsysofFlaorchersisl "Comropm Boiluisn] dtrsen MEnvrmens Laboratory 3M EnvironmentalLaboratory Page 180019 Pose sot226 Page 46 CHAT Report E01-1245 Repon E0r1245 PROS: A Reproduction Study with the Nothern Bobwhite PFOS: AReprStouydwihuthocNortthein Boobns SDuilaution atAmayt: Box Nombre tachment: DikionsSummary Worksheet SNEavxtreTunicymoTenNpDNiuenmbatenr:a: EL sampteNomber [oie| orton [wv TvTvTvTo TwTw[commons [Verieany:| I rrrrrrr -- rrr rrr rrrrr rrrrrr rrrrrr rr r rr -- rrrrrr rrrrr rrr rrr rr rrrrr rr -- rrr TT 1 L Nore: rT T TT 1 Vi0diion= ___ofsamples ____ofsvent Form Completion VertiedBy Soi rn EnictioEn osdsAaviisof Forces Compounds om romp Nae Fae 190019 SMEnoomentaLaboratory 3M Environmental Laboratory Paa7gct2e26 Page 47 Chaz Report EOI-1245 Revo E01 1245 PFOS: A Reproduction Study with the Nothern Bobwhite PFOS: ARopiStudoy wdihhusNocrhatmBootna ATTACHMENT B: DATA SUMMARY TABLES SM Envionmona Lavorstoy 3M Environmental Laboratory Page dncizzs Page 48 CRAs -- 2 f fanz z Hi 2 rier : 3 z Bs: i: i it ros se) eK g< 4 Hee oe HLac c 5f " HIER at | eel sas i ERNE HE aa . 5: I IK z BE | g j :| a: FKRees sas | iBe oo * 5s PH EEes suf PE RB : BR: eel zoel 5 o g ii 8 i? g[AN WARREN il : Bi HE ii E N;E HEE H1 ii idl: HE 2 z Hii E] iB <i 5 p Bun Biima:Pii :ss0: EPuEa PBelnui:n Zg 5 Halecdl sad + HHH ER g 23 3f :f nae xs _--_-- 2 axgsazliny HEH f2 Es 3 z |< <| | gadis) HIT E5515 i |< < << FR (RORITI PE 2g= i i Bl 1 iiRLi| 8 le RRRRR of H erzkrzlResl 3 ] 4 H f ifli es i| iEBl J1E]R: eases o JR Se Jdele J line 1) <dlria biiidseseiig H|H: F| HW OUR: W | [i G i3EMf il BfHeckIecGpsHy ER3 WlH aepiE; it : W Eic C EER h ol o3 2, Bi SE Ce go illliackackad a ills iladaail: af & 5i. 33 i =3 Chien ------ T 3 sH ys saaggaeesinsgtei : Rs i 2 << ease rasan < ify Beep EEE RERESESE Iz eslseseasisas| of ER! i 22afzaalrar(ees] f ffses- foecfess H.. 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Coarwvoeicmonmap ntnte2ro0ot1w0a7it00055r00o0ptmosCrBreALsa ahKnw2n.41T.,6e7Herd WES geod) 0ok 820% pa narsch eaeer-- m---- ------r hm e, croae Son amps wr neoussoa tr snc ctPsrtOoArtt(aMuortetnThC12ScIolEBF sep 15 vicstgime cre(23 _wotsi eel rite 3M EnvironmentalLaboratory Page 161 Chad le Pearl] ReportEOI-1245 SPE Columns Extraclion WERE roepionoe:ts OgnKasw 2793, Veet or angen | "ater a Settuhdy aRmevbiesiron:01ET152842581.0 Nae Sera [meme Jommomowoad gee [mee| wf |gee | [Consens [remeron] oebf| wn FE [me |eounonus fprmmomiicd gg fom | wm fete | tee | [eon [mr |e fev | wm foe | Foe | [oom om|i EE t me [oom [mm| fe| wf ree | [Coonan [oun | go Lrmar--| wpe[eeec | [Ceovoemn | Cro | we Pome | fo Toeme | oCreooen17|2 resomtar|t a1ve2mfom Fat ons emaeniseoasr prAersehooest 280 SETH ee saecare or20mi 30 mn ska iki L021 Serwit eet omen Tia | sowlne[we| re al2oivseuCcuttoiaug] : osLb ) boo1t. i OGK. 4 O@* 1202 n Cmasno essarmae mameoTnYe.n [9 Fiersamplethroughconditionedcolumn,discardingfitrate Eeeccommm eovacntsssnotesvnn ttaocsdsd TTSSII ospmt t1csnn iS Zadar "3M Enironmental Laboratory Page 162 661505 Repo E0124 recon x TOVAGR AS E ERE pron [mmm] ee | PrepDate: `Analysts initials: 212002 R-) 02 EN PL 2-1 on - MethodRevision: ETS-8-231.0 `Study Number: E01-1245 Matrix: Sera. msAvene sanpis| Sorpletomber | somarees | Toyt cpom | Amonudnspte| xensven oeem [eheee ofP Ee;aere| T,fl[aaoe | oven [errr fm | fi om | CTooe [e= ar mFr=e [|ow fEoE [oence || on[ea fm |, mr [CO] ee [ei fe| [hen | [oan[| fo |p r| ree 1 rtet al t J r ee t fee eerr ee bk rer oHamre r rSeEz en perna GH,J1 uddted ei mtr tes sabes oo p Ere ei oun WO Hsagsealiguciifer [Ctr oma monCeomneiat SFi aS omrm o :ne ue. amount ofsovert (Leh. es at 3M Environmental Laboratory oh det" by Hoo : ox fue0l FU535) SS ito appropriat1e5miceniruge 2 et feo mi oR Report E01-1245 PFOS: A Reproduction Study with the Nothern Bobwhite 'BOTTLE-TOP DISPENSER DAILY CALIBRATION CHECK Smeivroaeesr, VARs Sud [rwm r [va ee Date: 21:09 DiispmemIneeirtiaIIlDos;;: 0310K9J% Volume: 5 wil Solvent: ACN Wa oA 4195 [211 2 os 3 s 4 1S 1 os 2| [T2 1o39 k51 | 3 Ss8 o> | [aT294% | o3%57 |seat | 133 |o34 %f | 1 co7 er | |__VA olumv e:| eS roa gcge| [ %Accu|ra Jooc .3y 70:| [Sube:| --ocoszey | [--wev 01a % l Para |Biv] Cen fs =5 w sw0% ] Performance Specifications Corre = oe Haken. 2102 OK "Density values taken from the Merck Index, 12 Edition, Copyright 199 , =specific gravity at y*** referred to water at Ce o-Hexane d**, =0.660 Acetone d'; =0.788 MethyMleetnheanChollori*d*,e=40.7,8=616.3255 3M Environmental Laboratory Page 164 Goan? Report E01-1245 SPE Columns ExtracRBORoMkRRgrbduction PainBopyhic IE =e ee py AnJ alystsinty ial: ce 1c] SMtetyhoNduRmebveisri:on:c 7--o 2& 4"523.0haze0 omer foram fora [roams promo poroowe formasmn forma formaor formas ein | i | sober ogee |TTTT eesmTen | [empSIN TTTer meg NT 1 Tam TL [omps NT TTTeaesy|[| [ops NT leew [1 [omp eaR e]TO Joep SAN Nceny] TO omas TNT oe T [meg [TNTTlead]7 oreo foroasmn roam perma formaor roam [oops 1PNE TomeT 1 [omerNT eam | T [rego | KTfase |] emp TNTlowe| [ mgm TNT wag [[ [me TTTNT eel[1 "aus woe yin Sul, if oDoro frees [mR Dwg[1 proass [Tom INT fieseg| [| proase[mmee TTX leg|[1 ore [mop [1 Nees TT] rome Jemem TTfT a [VU] Aoruasgmeo.tbotra t 5 Amt oopAcenoonne vt355r R SSE oeassaarsise2i0sn0 Gi e B arsar)g w+ Cae rarn ritaicyyre irs aSespwiirvpeee ce oe. CVFiorenaaCroibuenmhoacHraocnhrWThkp,dcr CRicciogSoocnnsdanr7ts et Th) tospp 15a he ofammnton TL TS 3 3MEnvironmental Laboratory Page 165 Cha por B01 uo]248 PE Columns Exmchon wos PFOAS: Reprodu1"StUf SMEtheR A Biehit Ce fe de ea a prapoae: 27.02/24C2 Analysts initials: fH py phen] ev Method Revision: <3 ~ 237.| SMtautdryixNum1b0er: x=/- 2% 4S)eer teri Feiss] eo proses proawen pons prooaae proses reso poroawn oroeaon proses [opm TRTT TT ser | [omeSV RT TTlesen[[1] [om SRT TT ear Joma NTToT psy[1] oS m ST had| [empSTN TT feoany [| [vpS TN fess [TO ome R TT Netwg[T [EET ONG TT fresky[[ porvaswe [vm 1 Od llewaig[[| rrsesoas [emp SAIN Loma T [ ones JompEr]1 TNT liesesl[1 fons Jom Sl NT lowe COREE tsi IN AN NO NNI2772 I poss Jel 1 TNT lem[1 posse 0] TN lias,TT pram IRE 0 RN fey[1 proses IR TIN Jeane[|] posse JET 1 1 TNfase fT mee Jom]1 1 1 Trewn,TU] T-- emesho a3ArctsrtAotritu 5 TM Sepeetsaorviesscie15 prt he AdarSmaofgAH 2(T0HA Gr w1 austedsample 8o-_-- O= rca40 mfKarcaitohSeO= vipnveip a oCeamohn yHe t Coent erg Faret Cmooe i carroWie cnnhe Lieoar gooey A Sopope vertsasPo aT A esas Tan ap iserpedsti : Soules woe cighedad * ob howtoJqentiaed ks= fod. > f ok 3MEnvironmental Laboratory Page 166 [LEER] Cee| Report EOI-1245 | Anpraeypsonse;tial0s:3C:H2| [HoT52 spe comms ExnclPRakrcion Sg ana Bei MSettuhdyodNuRmebviesri:on:<13.77522931.1 Matrix: Zz "ol 2 Ec ro a] sereat fone [oem Toney| al forma formas Jom mr XT Troma Joma NT Hee[1 foes [ome NTeT es 1 femme onan JJoeN mm | RTTT TTreaagg|[TTTH fname [omen NT ese TT [prem [omer|X Nese [nes [emp RT rose TT foram Tem N Teese I TH rma [omemn RGTT Tiong TH foram [ompmr| TENTT1 oan] TT fname nese ome [omeS ENT eee,| [ TT NTTR ae,I froma [ome TR Too ens | ormasone [mmer| NTT Tend| [1 fporrmemsans| [epee p T TNN | | Tleressseg[TH fo[ rmas mNT C iow | T orsason: [mm| IN Tween] pose formas [moe TNlaste [| 1 [ome [T SeT ss| SPireammt 2 ko 15 polaa rmoatsts 15T7 ad PAroeAk<La4efh,c1.0 ApLlon HALT cauplep etapa hl waneAE" CAHMI REER $e `FShakeesmampafor2u0mnin@00pm Stnoaslairs_s__a_n_e) CaetnStoomnsaam ro 10tmin @S20o0m0op (CnoriSES ov aexrer5 o TRoa 1) meliatins i DeC ceasntaero act lpoto cyntHifaugen eCsoreeseavtehwelt. danke Foatticma oeennnc oa,e dacs e Sioo caomrggtiowhcetoemtpdetasan1st ofPsEhOaNnAk_a_.__Ththo poepreet_15 m_ic_asmyibeatin Toi amp iapprepave to 5: shoeuuwdeegsensa;apneed, GrbyiskHee3.ov!tib; eior.0 IM Environmental Laboratory Page 167 CHAE Report 01-1245 PROS: A Reproduction Studyif he qian Bobwhite . SPE ColumnsExtractionWorksheet Fo wets IWaka EEpToonreus ee Pp aOrHp ier |e | wai] Matador:<77.a . 231|.TP somes proaon [FnSNTT Tioske|, | fermen [mea NT seg,| roomed [roams [ mg Se N mm emea r|[| fonsaon owen] IN 1 1 Jee [| pos Jomo] TN 1 | Teewey[[| pornasom prose -- Jomg Tomo T TNT N TlA eeoenssT | |T [roa [Ewe ENT [me [| promos [emaor TT ONT eas| fproasen [mem NT Jemsa [| proses [Ewe INT ease [| LTTT NT Tw 1 1] 1 1 1 NT11711 T 1 T [1711 r 17 T 1 IN {111 11 TN TTT] 11 7 N11 ITT 1 C1 1TN [v1] FSeoentnvoetieboent v Troray tmearrottan 7 rd pSS rotaiseemtpeprecarryy e torsseane 4~ COoKnncotirwgetaanhenfe1t0o b22e00m0m (Coe ze Ce ootanpm eosooya TE FfarssetgISr eesec Gse,minem, TSiroames amonwii mSnesaPEOAssoRT fu se, angles weeweisghec!od by ov 4 oes ted by HoT, fo? 3M Environmental Laboratory Page 168 co1iTl Ce le Fearaman a Repo- s roaens aDgEfti? prey come TAR PFOS: AReproduction E Syud TBobuhi iStoerA37.1 2301 i svercinss oo proves [PEF 7TT fomeg[yp | ooo Jem TT Tee rrnenm omemT Taew[[ roe TomemRTT Tew?[TT [rNe T m hao e,TT fforomsrse [oomme TN KR hesns, |] prroeaa Jom IN TT edeag roam |e TT egeecy| frre mam NT agen| proses wml NG] eed prone Pomp aga [1 poms fone mem NT feetTT Tom sel 1INT Jem | T1 proone [Pie 1 1 NT losses][1 sone "om 1TN Jews 1 ] poses PRT TN desvg[1 1 1 INT "TT 1 1 TN1 141 OIGN, ghd out bot, heausgeusboy #3. BanekgeorsToAt AwAomnousnvt:t 5 Trt|ASNanEtSeTe w.e1stAnwdeaARaEndimiSTsr PRbY m SaASimekssanroonbeidoy1SmipprcJpeaneibe Poco ar Soumptian 6.35 - 23m Foo(n=F:D ThWIeZboEa@i CSPoaeiTa atroe201401GiRO@Sosow Co485 ee las EGueesAsL waeTesE Trars tcica Srlyste ese soe rwge cur fA : iN jaTis een Ca oFnadirtsioanrceohPaemnnwhiohMnreiOHre( seA ,d)co GEtRrTFo p~3L2demme AlSloewccoloumnmtomgooconmplsetelaymdory. lst____Te) log m-oFroemshoe Tari amon i sToe ve ort `Sphesampfeswithinternalstandard PFOAstandardnumber conc. amountadded. Souples aud i were wigpi ve Jor souls' i ju Fie < 3M Environmental Laboratory diy . Page 169 001152 Report 01-1245 PE GotanExes gardcin Sly ation Rye Eo Ta Vat po] AprneaplDyastte:nals: O21K2H0O0J2 01o0k samen aay | isa sn pornasen|ocooms| iow[uw| fornasoo Lomein| oe | tan | MeSttuhdoydNuRemvbiesri.onE:01E-T158.482-0243316.50. 295 @Dadd le Matrix Liver wm |mem |wa | ow |maem| [orm | rememisars| as |"ite|sosoromoes| wom | || [proses [FrmTy | ite |ousoemoras|masm| || fesse [Fm | oie|auorcamnes| meaom| || [msi [m | ter |swocames|y mom ||| [rns [Salmo| iy |"iter | sousommas|mom | || oraz [FE]py | be |osama| Tt || [roe [SEE | Toit |sooo| menem | || fini [FEE | "tien [somone]mom ||| [presen [SELB | Toi | osscnmies| menos| || forse [no py | en |e] mono ||| [reson [omni| ge |ver | isto |maz ||| Jorn[FE Ty|as| ssomonso|mono || [renee [Femme | tie |msomons| maz ||| Jerse [T |Te|susan |y moron||| [rneses Frm ee oe |TN | sons| Tas |SR AST KSiHloavaestmotThHotAetwSiighIKD1ae5rmanpnAamwboeautoenrptMlmeeeenrtne5f.r235 6ieldn_k45vmeir THA ASr1c4unt_twAtmroundtioedfr,91.CimS" SeiataamsmootsrOnscNaArIyLS tositessame Ceorreusgaempample2f0om1i0nG3@200o0er(CeUnRtosw24peGYSDh OCH DAuca4t0rmaatKalnyotct ae50stipneetrpecoi oe Caotneasnasmoeesmigh MseOrHr(HrAose51mi5n4g FairsaCmoplae oughancdonydonWeadaslo, acd te EASiooowncc soaommneo tnoigocm camomdm pssaemnio8ig PsFs oOAnst (rHECrirC, 2S1514)45ris aoer1010o 53mcgrstvgmeeousne isos 4: Tarr ari i serrata es tor - b de Eubine exhachbou deve yf ox. {foRw 3MEnvironmental Laboratory Page 170 [EEF] Report E01-1245 PArna\elnypalsDytsasttsen:iiainl9tials-:1! 2 0 scons cxscrPFeOmS:moAnRmepernodu 8ctig nStuaT a L dywith je Nothern Bobwhite MetSthuodd2yRNNeuuvimmsbieornm:: e(o07onS/t---)294-2AH 2r33-oA 1ar-E0) EE enum pornsen| ocoorms| aa | Em |waonos| Stim|[| poroesen| ognem | awa [Em Lsosaomose| Soto | 1] fborrooesron||ccoommmmss|| woex ||HmEEn||oowveaommoons]|BMomom ||1 || fporronmoes| | cecqompn || owew ||"viemin Loeorvnenna]|fSomom||[ [| | fborrnoemsoen ||omcommipns|| wgens [|"TwiEnLoosanna|]mSomomm |7J fren |omarion|e [TE [mm 1 bfrrooomns|oommareionn||epe[ |CE |w| mmm[TT1]] former [omarion | ga [um| ow [REST] formes |omen| ge | im| ow [AECL L f[orrmmaesn |[oommmaoemiroe|| pe | | um| ow || wmmmmm[ [11]] C--=rr =1= r 11 r a1 r Blank ver TAS 1475 Amoouf m ver:_905593 Blank LiverThA 5 142);Amouofnivetr: LOKo `Homagenize iver withKandiyohiwater;Amountofwateradded _45_ mi `Amofowatuernaddted__ mi ERSr r etereeeetssera r mmrsepoiea--o r--iosno nn oonpehro3no41o0en03 Conditioncolumn with MeOH (TNA.5154) TA reomemar--aepan--rene----cet, CE Trantersale icapprorately marked autoPr obarbeu slot be on.oCcE". 3M Environmental Laboratory Page 171, CoLisg Report E01-1245 Ero Tomar: me ERE =e PFOS: A Reproduction Study with the Nothern Bobwhite: EEE mans mips peo +apot a EEE(ElE[E[E] BE ETETE) EEEEEE || E fas | E Colada hed cased To doyws Hauke Ail =A 3M Environmental Laboratory ee a ln Page 172 Gains Report 01-1245 PFOS: A Reproduction Study with the Nothern Bobwhite BOTTLE-TOP DISPENSER DAILY CALIBRATION CHECK Manufacturer Vou & and Model: (. Volume: 5g TO a 30Niall IDE: Tov 4. 914 H2%zr lode T7550 | ET 5vw I EE 30577 3 5eey TT e3d | Cs TeT s f T8553 | [Av Volue me:|ra 077g % e| [ wacourJa og.e F y:| iet : [v CC cv08i 37s l lesplancern Chri FT] Ce oso | whew | Nove Yates B=) Zocor "Density values taken from the Merck Index, 12" Edition, Copyright 1996 d, =specific gravity at y** referred to water at xTM n-Hexane d**, =0.660 Acetone d'*, =0.788 Methanol d, =0.7866 Methylene Chloride a, =1.3255 3M Environmental Laboratory Page 173 GHA 5G Report EO1--1245 SPEColumns ExtractARS nReprodEg i deApter Baebwwhis eT Tere Ee] PArnepaDastet:als. a 2O1K2H2o0J02 2o1u2.02 Mpethond RebvisionN: ETSi-8:2a31.0,s, ; JY Matrix __ ver reer | nso | vwaoraio | soke mcnea] vous fone frase meso | remem| en |OlToma be TT |remeesoma| oe | gg loool wits [1] |omnesorim| wa | vane |ow [om TO] poriasons |ommesonon| we | aww | ow |ostTT foes opm | we |wm|ow |mS TT formes ommesouie| we | em |ow |iTT promo |ommuserim| wn | vem | ow [aii TT] CIEE peeve) PPR kul ITS NE I ronsoe |ommeserim| we | wm |ow aig TT] [reson |opmusoner| ws | me | ow |ono [T fornsom |ommumorom| oe | wm |ow [igs[1 fone omusiie| we |um|ow |ome TT foes owewsmrow| ee | mmm|ow Lowi TT] Ble A51D Amnotver 5:03Dg _ parker THA:SL amountorwer 10'S `eHoomogernitzetiveerwirth K1a6ndmiyiohpiwaatebr;oApmoauonatnofwateradded iS mw Amooufwnatter added __) m CeotnemeataACoe(toAaroUinS@ omorasirotetdososianSe j00641169>4 Oanc1e0rm atKalnotcenotgee e50s ipwprp cot 2s VmiaCralunm weasnodmTWmahteatr SoToSn) NFoerwscaomnttvaegohcocmoprtdeone8d7colon, dcreg rte . tecomwen croreed amounntpofrsoovsent m(tHErOH uCa.SITUEE)S Ltospeeoepae160m-iSerptsmtrinaSl Tarr aml io spprareatxeyhraecchdotuteours daw byal or Oafrro 0 WY 3M Environmental Laboratory Page 174 ALERT Report 01-1245 57 comsExincu OSH BEPSCSh2i42eh Bh Bobs po gm on AS iy Foss oas: okros 5% Lo Sty Naroa:Sos 45 07.2 r=]Taree] oo py [meson [presse piers lost fritid |omarion| en |Chg | ow |nha [omnes | we [euamFw ame Comments ug | oman |omursmrow| we |og=lw |omRECTT foram [omen| we | m=]ow |RESO IT] forme ommsoroe| ee [oulTw|eTT forma emerson| en ong =e foresee |ommearow| ov |ObsiFTw[ameTT [pres |ommsovi | we [enammw ame TT frmesoer |omersriw| en |ohpiS we ame TT [rene |ommsmwoe|ee |enamlw mm TT meses |opmusrim| ov | hee | ow |mum [TO frien |opus | on | gre| ow [om TT formas |ommuserue| =v |Tope | ow |mm [| F [ F rl F + [| rT TT IFe TT rr LL --4 - t----a +11--p1 11 11 TT 1 1{ | thachou dove by ow. ihe BlankLiver TNAS'4_; Amooufivner:t 5-0>3g5 BlankLiver TNA4Z2 Amouofnvter: 1.015 _g. rr aisin tn ien `Homogenize IverwithKandiyohiwater;Amountofwater added +S m `Amofowatueranddted__ 3__ mi ing mi85007 rae OR rp R 3) 0418 OCeevneetHrttougmeetrsatamptlreedfeoem10tmirnu@ 52e000wl0pimp(rConntogeAInHN16he.61D aesome SA------TETY `CoFontnditsiGoanorcoeheen whiKtahcnMieoOnHe(Th-cA:_S7ad5sc1ri,e Ri soem cmt 1 . Ttiectonaanrweihhenesscmapedamamoyuantmonafrets irsovent o(HnOmH o1asAiSo TnSAEorBrtsspeoocpttemreS1165g.mgdcaonet mmooinretd Jal 3M Environmental Laboratory MES OFAAK | Page 175 ALLGS Report E01-1245 PFOS: A Reproduction Study with the Nothern Bobwhite BOTTLE-TOP DISPENSER DAILY CALIBRATION CHECK [re] [eg] Manpurfactouwreer yf Gand -12-03, BareR DiBseptaeTnnoesteeaIrlD;s; 150%70,9% VSoalvTuaomnset:: ta5rcsohenl ara [519 o3sy3 | 3% %9 | [ [C C3o12>a91 si3oaqsg[ A 1 --|T | S5= oe5e0sr ||| F [Av Volue me:|r5.a c3g 50 e| [Sade vozes | Pasha] BF] Performance Specifications eolT ) weBhen Teonw] Corrective Aon ove HabeLR [29] "DensityOLvalues kenafryom he Merch Inde, 12 Edin, Copyright 199 n-Hexane 0%, =0.660 Acetone d',=0.788 Methanol d*, =0.7866 Methylene Chloride d*,=1.3255 3M Environmental Laboratory Page 176 Chains Report EOI-1245 PFOS: A Reproduction Study with the Nothern Bobwhite tl Sn Jug SPE Columns Extraction Worksheet fage Voi)3 otlaila APnraelpyDsattse:nals: 2O1K3H72O0J02 samp nomen adescr | rnc maior | - MSteutdhyodNuRmebveisri:on:EOE1T-S1.284.5231.0 Mor Sera ea | "vou Comments bosses Jamsso | wwe | wma | ow |wae | losers Jounsos | ae| mm | wa JW] Jervisecerss Jomus|owp o |mwnasm|smewon n | s me ||| lease Jspunums msone|naamez| vaso |swoon [Her||| lerissers Jaswsomsonea|naees|"vi|tswooonr [Baw|[| Jesters nusrgesos|e|"omeLoving|mw|[| Jovinisons [soussooner | vs | vit |oo|wWaot |c| om] se [ws| tn [ovoomosa[Bei |[| ------ 11> TI 1 1' 1 ~1 117 | 11 = 1117 1 1 1 ~~ 1 TT1 4+TT 117 1 CC Fae | | TT 1 1 1 [1 11 1 -- 1 = 1 1' [11 -- = 11 117| c rF [ r e t1 -- 1 T1 TT [ [V1 v 1 ]1] ere Sark rsera A599 2 of setaan tit 2 1 uSpoetsnanoessaccirson1gy5m pavpropone ie A 5omfAlCN (TN-A-4145) to alquated sample `CSahank4e0tsaumpoaflKemfolra2fo0rmhi1wn0a@tm3n0@0 r20p050m001m(SphapkreCorewpreyultinA nse2co_20u4sIGUAbEeY.12) nOnacentseamxpalcetiighct fno atesepsopweinmwnar `WaCauonsrdhitsCiaolnmcaeonlruwmonitwuiKthaaMnnyedoOen(rWTaetNecAroSli1,5S4e_c)on rte Nowour tog ampldyy itecokumnwith i amountof sovent (HCH va. S754) intoappropriate 15 mi cantrtuge ut Tarr same ooTH seprxaybmaarckehdiooinl dove bX ok 3M Environmental Laboratory NA) Page 177 NLLG0 Report 01-1245 PFOS: A Reproduction Study with the Nothern Bobwhite Bottle-Top Dispenser Daily Calibration Check hor CORY OF ie"2 rSoraisiNAL Eo Niamatactorer sa Mode nical DiBsapleannsceer IIDD:: ValSumoeve(nmtb:: Ds: nemo RST Sep 0510909 Aosn TAs Ke @ Massof solvent (9) | Densmibty' | VoSlouvemnetof = 1T smwsToomms ||ssoows || T Cs e 1 sow 1 w e[omwss ||ssooss|| Cs 1 som | ows |sow| [FAs TOG] pass Pass Volume, Accuracy, Performance Specifications cv. (ml) 0 on Ce 1ssoiewe1e Siow Correave Aci: Neve TREN of 2120, "Density values taken from the Merck Index, 12 Edition, Copyright 1996. @, =specific gravity at y*"*C referred to water at x** n-Hexane d*, =0.660 Acetone d*, =0.788 Methanol d*, =0.7866 Methylene Chloride d*', 1.3255 3M Environmental Laboratory Page 178 niga FE TETT EE E Report 01-1245 TT Analysts inals; ~~ OKIHOJ pe rE Gots ExacioSRTHMBAPWYoSoCdTRSoLoof DS2158f2s5N10 e Dobie Study NumbeMr.att01L-ive1r 374252a:e37.83.8 | ventair |aov eaa formas Jomo| ee[ge] ow 1opin TT) forse [omnammon| [rm [mma | [prams Towmamerow| [mae [emma | fons |ogmuomrow| [rams [emesmmon| prison Towmumroe| [prea eomesmroe| oe [ota[ow ole TT wn [hen[a[merTT ov [otiTw[ae TT oe | ote TwPamirTT] oe [Sham wm owl 1] ee |g wa TT] oc [ota [we |omle TT] en [opmTeTrTT) Svar THA_S103_ Amoof eur_5n02t5% Ba vr T5142 Amountative 100152 AHe roemogevniot zetlieverrwn iithsKTanodmibyohipopes water, Amount of Fi wateradded_48%S 4 wk `Amount of water added _9mi rRe emstaomfplReON (20m1i 43050m (tSheakerV2I a) R CUE Cian1u0tme aKmapya10tmir@ G2O00p05 l(onrecHnfXE e Oecnt r exchsiogcarnastieesrwinwate oConnoCnocteoiinarMiAOtHe(TrA121) S Foursome brash consort, Gscardng seN Tantra lesspaosrtlnyemwaersrd staorts arr EIS L t ogt amaumssnd_Zie exphcTion Done BY OK (ho 3M Environmental Laboratory O@ ok IY Page 179 nas Report 01-1245 SPE Columns. ExactcRRS BryfnStr bdedistor Bobwhite CeesTeele] APnraelpysDiatse:ntials: ~~2O1K3I2H0O0J2 pie MSettudhyodNuRemvbiesrio:n:EO1E-T1S3-482-/213316.50 Mat: Liver edi | wasn | pk mone] vou foraons [omnes | ov [Cha ow TREET] Er ERE ever BENi ET e a) ial TS Ie rime |opmesemun| ov [oo | ow [rs[1 promors |omesworow| ot | hppa |ow [ibe TT] oro forason Lommumeon [we Vogel ow |eTT Lommumnoe| oe [NgeeTanTT fron fora fra |omm| ao0m[motin[ow LambsTOT Lommomrie| on [pir] ow Pais TT) |ommwn| oc [opt lwLie TOT] 41-1 I | -- 11 rT 1 11 I 1 CC -- fF Twa | TT | --t------+----+--[ + {-- | HBloamnkegUenreicAv8r w1ih4KanyoAn wmateur Aeemeto_5oof03v5aft3e cded_4BGEi3rwkEoyIerTA__SA1m4o2ur tAatmeurnaodfed_Sivmelr_100150 ASpiicnosamofswcscatroan1g5y. poeple aon 5ShakmesaofmApCoNr(20Ami4n1@530)0psmuSmheakdersiampsle<p 6% C6o6n4t0emoafmKpalnehfyooht10atienr @to2500070mopry(rCeennteecHarEr1i1e01oe7. DeP cantp stac--c-- eas wtih--voar WCoahndCoasmhewniiKhanMceyOoHn W(aTtrer.1654) Fitesanph though connedcoke, discarding te oeceomr saecsensmsoonaart soonmseHmeeCHaCB ,02S154) sod pB15Rm contig bese Sh ransier sre to spate mated atv extraction dove by ok. @ of 9 gg 3M Environmental Laboratory Page 180 GOLLBTD Report E01-1245 PFOS: A Reproduction Study with the Nothern Bobwhite Bottle-Top Dispenser Daily Calibration Check Framutacturer 3nd Mode, Date: Initials DiBsapleannsceer IIDD:: VolumSeol(vmeLn)t:: i: VR Sl pene wm py oc fe) 099009 AcsN NA Massof solvent (| Dengsidty' VSoolluvmeentof [ Te n 5s n T1 omoT mssrs | | ssooas || [ C s+ T T 1"ssseeoem3 mssT| |oommomsser7 | |Issso ooess||| [mgs foe] Pass [S Deva t 0169 | Pass [CPCass/FaPiAStS ]| Volume, PerformaAncccueraScpye,cifications cv. ml) 0 00 Ce wees| e iow | Corrective Aeon: Nowe TAKER ' Tok 1308, "Density values taken from the Merck Index, 12* Edition, Copyright 1996 @, specific gravity at y*"TM referred to water at x"*" n-Hexane d"*, =0.660 Acetone d*, =0.788 MethyMleetnheaCnhollordiTMd*,e =d0.,7=816.63255 3M Environmental Laboratory Page 181 Cots ReportEO1-1245 PFOS: A Reproduction Study with the Nothern Bobwhite Achmet D: Dilotons Summary Worksheet -- Sen Number: OWS WIS PED D[ilRutioonkDaite/Antalyest:n(0 i Extraction Date/Analyst: #033 coz 2/CR) orPeseipton [uno[noo[vTw_Tv[uu]Commens [VeritedBy:| [Eorisanso |jp[x[FTTT [pada |E01-124528482| HHA [Eoresze [x | Ab [oozes [11x | 1 1[ [7/7 | | [eormesmas| TF [eormesmee|TTT /T [eormeszsir||x[TT [eorsmoe[|x TT|Jolla ||| (pormesmes |[x[TT] (ormemer [Tx fT 11 | [ovmeszses|x 7 [((r[ o[ Eoormm an[| | x| 7[|[] |] (forrmmess [|[7 xx|[] | pI emrmms s | x (| | [| | [Nrore sa[vy 1100ition 101ofsample380! ofsven Al iuwdreomadnonsdayof extraction rte dn tesmi. 00 4 462 GED), o ] Be om ComplVeirtony: 1h Gsteery 3M Environmental Laboratory Page 182 GHAR Report E01-1245 PFOS: A Reproduction Study with the Nothern Bobwhite: tachment: Dilutions Summary Worksheet BDsowixaeNynmaDaErtsnAmsEyobs1:34E2 o (24 G03 4+MSEareeiacTinmoNnaNapumatb,erm:GM:oesOIe2IS1r43S1?3WIS PFDA Sample Number or Deseripion [Enna [[ x tT pid) [pwsaseo[x | | TT[ZT] [ewes x| | [1 [ ~1 1 | [ewsesms||[[1 7 |[1] [owneare x| | | 1 1] [onewe x|| | | 7 [| [psa ||[| [4TL [pesos x[| 7[| [ase[x| | [1/1[[|| [osesmes x| |[AT[1 1] [posasm x|| [7 [1] [eo[xv|s[a7s[o1 [[11] [posesow | [] loans[x |[TTT 1] [pores = |71[TT lesan[x[7 | | | [ [ T [] [oan[[ x TT [oNo3t0sdesi:lautsio=nr1[00xu of[sam~+p9l0T 0eulafvF ent ----------J|| 17100 dan 10ulof sample 990ul ofvent Alans wre maconayo extaction otedn teats. OKT Po B25" Form Completion Verified By: E21 d-cvov2 3M Environmental Laboratory Page 183 COREG Report EOL-1245 5PE Columns ExtacFIRSEnBapr RSIS Bobi ros m Saosnym WBA MSetyhoNdoRmobviorirnE:OrEST5-8:S2910D" 2 443 we Tl, Cr Jy Er -- [oosrmmeermoerrts]|we| [oe E EERE free] EET [oman|oe Tgw] m |Sif [1| [oromme Tommeworuw| we |oyewv m |i TT poison ommuomon| we PREGow] Toad TT fprrnmon omneieriw| oe [newTenTT Pommecron| oe [SheBT |BEET] C[oneT sies TCommrercyoum|I oc [shil| TwmN[oe i [1 | fpornesion Townson | 00 [s| wm |ooi TT frien [opmaseroe| en [TpgTM|ow Taf TT presi [ommermue| oe |Tug] wm [RRS TT [o onswmemor | u oee [hase| wm |Te T sE rior ut,_310T _-- shmett,_SG atsBn Tk_812 mteatos_L1180 orermea emrm AoEn tyoewravntas3) I rm eeE ----E --. Ee fTem ewompe------ do Condtian column with MeOH (TN-A- 5754. Rowtonpamreasnoonotsaves HOH ur Si.741 apt1s conti LtHeA coat maA sFONmars ODE 3b5e cnor Eeo n son Extachon dove by of Ww {go 3M Environmental Laboratory Page 184 COE IE I ry ee Ee py Report E01-1245 TPropbonse : is oasezs0.02 SPE Columns 259; % ExtactioRRhRapSrtoWeugldshytodiNaRoneoSvortlu,opdny01E1B 2T9 .85b.248i10s0tS6h0e055Bonbbwah ite 2.15 oK Mate i Li [orm Tommi| we [haw[eT frneos |ommermui | oe [RSS]ow [omerTT froromevs e Toommummuoiw|| ooeo [[SmB=E] ] wwue |TGoe E 1 [[rnreoonse oommemrumssrmoum|| oooe [[hisSe|| ooww ToReEsSTTTT] rF os Tmo mromrin|| eeo e[[RoEeEms wR Ee TTTT [rFroonmms Toopmmson | omoe [[STheeae[|owow || E GEEET [fornoe om Tuommmmsmerne]| owoe [[hTmige[ lww Toa it[T1T]| FI --1-->[1 1 11 1F111 1 mama] [1 | rr ------ 1 1 [1 [1 1] [----t--[1114]1] J I em------ tdi,Sor TA fr mno t _0150 a Semper AoSddeeSmirsoofeAnCNfr(ToN-A-._i4G1g45)nTroE1rsnteie ustRedSsan mHpleO1n E4R) r ee --m -- : TtrEeEm--m----r----------T3 EossissnsimmrnerR aMet 21 re tm Se ---- exit doe by 00 ETS , J" KER f200L 3M Environmental Laboratory Page 185 Cares Report E01-1245 PFOS: A Reproduction Study with the Nothern Bobwhite: Bottle-Top Dispenser Daily Calibration Check Same TORE NE Dnaet: 215o2k002 DiSspaennseer 11:: oSom VoluSmoev(eanyt Acsx ph Dkdies Dey] Vater CwwweT Tooww[[saoens]] T T o w oom ww [|sosnoiw]] C Tws [[TvRoasnme]]|ssweomoi]]| ras [i--cor v m pass [Pasa]PASS] Volume, FerTormahneoecSp,ecfctiions cv. oy cn on Cio 1ose-iewe | om "Density values taken from the Merck Index, 12* Edition, Copyright 1996 &, =specific gravity at y**** referred to water at x**** n-Hexane 0%, =0.660 Acetone d*, =0.788 MethyMleetnheanCohllordi*d%,e =40.,781663255 fy ii no ewdoe ivd Jot dnecllyu towouder Was avaiteste 3M Environmental Laboratory ise P7UP A 1 9.0|0 Page 186 Cong Report 01-1245 Repongor 126s PFOS: A Reproduction Study with the Nothern Bobwhite: FOS: ARaphi Suc wih hoHono Sob ATTACHMENT DEVIATIONS E: STUDY PROTOCOL, PROTOCOL AMENDMENTS, AND aMEmon Lavoro 3M Environmental Laboratory [---- Page 187 CAHALD Report EOI-1245 PFOS: A Reproduction Study with the Nothern Bobwhite WILDLIFE INTERNATIONAL, LTD PROJECT NO.: 454-108, EOl12es AMENDMENT TO STUDY PROTOCOL STUDY TITLE: PFOS: A REPRODUCTION STUDY WITH THE NORTHERN BOBWHITE PROTOCOL NO: 454/120700/QRISUBAS4 SPONSOR: 3M Corporation EFFECTIVE DATE: January 24, 2002 AMENDMENT NO.: 9 PROJECTNO.: 454-108, 01-1245 AMENOMENT n"eTcheisssaamreyntdomceonmtplinectleutdheesaannalaytttiaccahlmpehnats(eAotfttahcihsmsetnutdyA.) which provides the information REASON: "The protocol did no include information for completing the analyses required by the sponsor. AMENDMENT: HaroldJohnson sthePrincipal Analytical Investigator REASON: "Thesponsorrequestedthat aprincipalanalytical investigatorbeaddedtotheprotocol. AMENDMENT; The SM Enver Lahrsory Pej enifcaion Number to be included with the study is REASON "The sponsor roquestedthtthe 3M Project Identification Number be included inthe protocol. 3M Environmental Laboratory Page 1012 Page 188 CaaLvL -- oRYoJ]GHEWOLN1245T25FANX M410 822 E0N83I2ZONBNAL 1vit42o33E 5 08 A ucion Stwy wsiththepNoather00B2obwhite | WILDLIFE INTERNATIONAL, LTD PROJECTNO; 454108, EOL-1245 Saait. ef,SadyBulb. TTaebosraier.y Teratins] 0 f HarEa 0.Job(neeCn.s ] ~ re-- `3MEnvireamental Laboratory, = La Led Be 1] 9fen 13 T Du 4.4 02 Oglosln Date z. 3M Environmental Laboratory Page2af2 Page 189 enn Report E01-1245 'WILDUIFE INTERNATIONAL, LTD PFOS: A Reproduction Study with the Nothern Bobwhite PROJECTNO. 454-108, E01-1245 Scan P_ Gallagher, Study Director Date. LaboraStEorByeMaenangeWmeintl ee TD Dw FarToon Pr Ai ee Dw "William K. Reagen, 3M Environmental Laboratory, Date. Laboratory Management Reha Foden, Speers Repsenaie Dw 3M EnvironmentalLaboratory Page 2002 Page 190 carne Report E01-1245 PFOS: A Reproduction Study with the Nothern Bobwhite Protocol Amendment # 9 - Attachment A Wildlife International Ltd. # 454-108 STUDY TITLE PFOS: A Reproduction Study with the Northern Bobwhite SPONSOR 3M Environmental Laboratory `Building 2-3E-09 PO Box 33331 St. Paul, MN 55133-3331 PERFORMING ANALYTICAL LABORATORY 3M Environmental Laboratory BuPiOldBinogx23-333E3-109 St. Paul, MN 55133-3331 LAB3OMREAnTviOrRonYmePntRaOl JLaEbCorTatIoDryE:NET0I1F-I1C24A5TION `Wildlife International Ltd. 454-108 3M Environmental Laboratory Page 191 CHILE Report EOI-1245 PFOS: A Reproduction Study with the Nothern Bobwhite Sponsor Study Identification PFOS: A Reproduction Study with the Norther Bobwhite 3M Environmental Laboratory Building 2-3E-09 PO Box 33331 St. Paul, MN 55133-3331 Principal Analytical Investigator SAtNuAdLYyTLIoCcAaLtTiEoSnTsING LABORATORY ANALYTICAL TESTING LABORATORY H3aMroElndviJroonhmnesnotnal Laboratory Building 2-3E-09 PO Box 33331 St. Paul, MN 55133-3331 651-778-6479 3M Environmental Laboratory Building 2-3E-09 935 Bush Ave. PO Box 33331 St. Paul, MN 55133-3331 PACE Tier 2 1700 Elm Stree, Suite 200 Minneapolis, MN 55414 Proposed Study Timetable ANALYTICAL EXPERIMENTAL STARTDATE ANALYTICAL EXPERIMENTAL TERMINATION Dare 01124102 03124102 3M Environmental Laboratory 2 3M Environmental Laboratory Page 192 CRATE Report EO1-1245 PFOS: A Reproduction Study with the Nothern Bobwhite 1. PURPOSE/OBJECTIVES Innomtihneail ndofsepshoafsepoefrtflhusorosotcuidayn,eNsuolrftohnaetre,Bopbowtahsistieuqmuasailltw(ePrFeOSre)guilnatrhleyirexfpeeodseidnotroderto epvuarlpuoasteeotfhteheifsfeacntasloytfidcialetpahrayseexopfotshuerestuupdoynisadtuolqtusaonvteirfyaPfFiOveS-mionntthhepleirvieord.andTsheera of btootchocmopnatrroelwainthd tdooxsiecdolNoogirctahlehirstBoopbawthhoiltoegiqcuaaill.resTuhltesafnraolymtitchaeliinn-lfiofrempaotritoinomnoafytbheisubsierdd study. The in-ife portionofthe study was completed at Wildlife Intemational, Ltd. in study 454-108, 2. REGULATORY COMPLIANCE "ETnhviisraonnalmyetnitcaall pPhraotseecsttiuodnyAwgiellncbye GcoonodducLtaebdoriantaocrcyorPdraacntciecewiStthantdhaerUdsn,it4e0dCStFaRtes792. 3. QuALITY ASSURANCE `sTthuedy3cMonEdnuvcitr,odnamtean,taanldLatbheorfaintaolrryeQpuoartlittoydAestseurrmainnceecUonmiptliwialnlcaeuwdiitthanGaolyotdicaLlabpohraasteory PPrraoccteidcuereSst.anTdhaerdQsuaalnidtywiAtshsu3rManEcnevUinriotnwmielnltarlepLoarbtoarlaltfoinrdyinSgtsatnodatrhdeOSpperoantsionrg, Representative and the Study Director. 4. REFERENCE SUBSTANCE 4.1 Physical Description Perfluorooctanesulfonate, potassium sal from Lot 217is a white crystalline material 4.2 PhausribteyeannsdhoSwtnabtioblietsyt--aTbhlee piunrimteytohfanLoolts#ol2u1t7iohnassfboereunpdteote6rmmoinnetdhstoatbeam8b6i.e9n%t. PFOS tIenmspterurmaetnutreDeintetchteivoanliLdiamtiitonDerteeporrmtinfartoimonstfuordyLPCS/OM1S3,a"nMdeGtCho/dMSValAindaaltyisoinsoafndFC807 `CoCnotmrpool,unadnsdiRneMfeetrheanncoeld.a"tRaebfaesre tinotthhee3CeMrtiEfnivciartoeonmfeAnntaallysLiasbo(rsattoorerdy)infotrhefuTretshte,r purity and stability information. 4.3 Storage Conditions--The test article may be stored at ambient temperature or lower. 4.4 Reserve Samples--A reserve sampleofPFOS from Lot #217 has been frozen at 20C 5C or colder and is keptinthe 3M Environmental Laboratory. 45 CAS Number--The CAS number is: 2795-39-3. 4.6 Molecular Weight--The molecular weight is 538 (CF1sSOSK). Note: The molecular weightof the anion (PFOS) is 499 (CaF1,505). 3M Environmental Laboratory 3 3M Environmental Laboratory Page 193 Report E01-1245 PFOS: A Reproduction Study with the Nothern Bobwhite 5. CowrmoL Matrices Typesofcontrol matrices and their source, physical description, storage requirements, and traceability numbers will be recorded in the raw data and included in the final report. 6. TestSystem Thetest system is Northern Bobwhite quail. The test system was dosed (at Wildlife International Ltd. during the in-Jife phaseofthis study)withfood prepared by mixing PFOS with feed in large mixer in appropriate portions to achieve the nominal 10 ppm level. A food dose verification validation study was performed by Wildife International under study number 454C-110 (3M #U2723), "Analytical Method Verification for the Determination of PFOS in Avian Diet" Wildiife International verified the actual dosed fNooordthfoerrthBiosbswthuidtyeasnedrtuhme arensdulltisvewrilslambpelceosnwtaeirneedcoilnletchteedin-blyifWe iWlidlidlfiefeInptheamsaetiroenpaolr,t Ltd. and sent to 3M Environmental Laboratory for analysis. 61 NJuosrttihfeimcaBtoibownhiotfetqhueaiSlerleepcrtesieonnt owifltdhbeirTdepsotpuSlaytsitonesmand they are an EPA orfectohemsmeemnadterdicespsfeociretshbeecsatuusdey tishteoy ddeotweerlmliinneiaflPaFboOrSatworaysednevpiorsointmeednti.n tThheeluivseer: and serumofmating adults that were fed PFOS. Furthermore, analysisofthese matrices from hatchlings will determineifPFOS was passed on to mating adults' offspring 7. SOURCEOF SUPPLYFORTHETESTSYSTEM "The birds were obtained by Wildiife International, Ltd. See their protocol (study 454-108) for information about bird origination location. 8. SPECIMENAND SAMPLE RECEIPTANDMAINTENANCE `ESnevriaraonndmelnivtearlweLarbeocroaltloercytefdorbaynaWliylsdiis.feTIhnete2m0at-iwoeneakl,eLntddp.oianntdssehriapapnedd tloivetrhesa3mMples received by the 3M Environmental Laboratorywerefromthe following birds: +40 control adult birds (20 mating pairs) that ingested food containing 0 ppm PFOS 40 experimental adult birds (20 mating pairs) that ingested food containing 10 ppm P20FOfiSrst generation offspring birds that ingested food containing 0 ppm PFOS dSeasmcprliebsedwiilnltbhee mmaeitnhtoadi.neTdhferoszaemnpelxecsewpitllwbheenkerpetmiosvoeladtefdorfrexotmratchteitoenstansdubasntaalnysciesdausring storage. 3M Environmental Laboratory 4 3M Environmental Laboratory Page 194 Report EOI-1245 PFOS: A Reproduction Study with the Nother Bobwhite 9. SUB-CONTRACTED ANALYSIS 91 Allsera and liver extractions as detailed inthis protocol amendment wil be performed at Pace Tier 2, 1700 Elm Street, Suite 200, Minneapolis, MN $5414. 9.2 Allsera and liver analyses as detailed in this protocol amendment willbeperformed at 3M Environmental Laboratory, Building 23-09, 935 Bush Avenue, St. Paul, MN 55106. 10. EXPERMENTAL CONDUCT . "The 21-week endpoint liver and sera samples received (see section 8) will be extracted and analyzed for PFOS quantities using the methods listed below. 10.1 Preparatory Methods 10.1.1 ETS-8-6, ExtractionofFluorochemical Compounds from Liver for Analysis Using HPLC-Electrospray/Mass Spectrometry 10.1.2 ETS-8-4, ExtractionofFluorochemical Compounds from Serum for Analysis Using HPLC-Electrospray/Mass Spectrometry 102 Analytical Methods 10.24 ETS-8.7, AnalysisofFluorochemicals in Liver Extracts Using HPLCElectrospra/y Mass Spectrometry 10.2.2 ETS-$-5, AnalysisofPotassium Perfluorooctanesulfonate or Other Fluorochemicals in Serum Extracts Using HPLC-Electrospray/Mass Spectrometry 11. METHOD VALIDATION Method validation for PFOS quantitation in quail sera and liver was performed at Centre Analytical Inc. in study 023-054. The method will be applied to sample analyses without additional validation 12. DATA QUALITY OBJECTIVES DURING SAMPLE ANALYSIS "The following criteria will be met using the validated calibration levels and fortification levels determined in section 11 121 System Suitability System suitabilitywilbe determined prior to the start and at the completion of cach analytical run. Prior to the calibration curve and after the last sampleofthe run three (3) mid-level unextracted calibration standards wilbeanalyzed. The peak area precision and retention time precision and peak asymmetry will be monitored at the beginning and the endof the run separately. Thepeak area precision must be equal to or less than 2.0% RSD, the precisionofthe retention time must be equal to or less than 2.5% RSD and the peak asymmetry (fronting or tailing) must be 0.5<AF<2.0, where AF is the asymmetry factor. Ifany itemof the: 3M Environmental Laboratory 5 3M Environmental Laboratory Page 195 601573 Report EOI-1245 PFOS: A Reproduction Study with the Nothern Bobwhite system suitability ails, system maintenance must be completed prior to running a second set ofsystem suitability samples and the system suitability must pass before starting the calibration. 122 Calibration It will consist ofa minimum ofnine (9) levels, includinag blank with surrogate and a blank without surrogate, the lowest standard will be at approximately 50%ofthe LLOQ and the highest standard will be approximately 50% higher than the ULOQ. `The standard curveequation wil bedeterminedbyregression analysis using the peak areasofthe analyte. The accuracyofeach level will be verified. Any level outside 80% - 120% accuracy must be deactivated, and regression re-calculated. `0.T9h8e5c.urAvlelmluesvetlshamvuesta scohrorwelaatrieosnopofnsdeetgerremaitnearttihoann(tw)ogrteiamteesrtthhaatnooftr hequballantko. `Thetotal numberofstandards excluded my not exceed 20%. Ifthe preceding criterion are not me, the curve and sample set will be reanalyzed. 12.3 L`Tihmeiltoswoefr Qliumaitnotfitquaatnitiotnat(iLonOQ(L)LOQ) was determined to be 10 ng/g in liver and 10 ng/mL serum during the method validationasdescribed above in section 11 The LLOQ may be determined to be higher than that found in the validation study during a screening to be done for endogenous levelsofPFOS in the blank matrices available at 3M Environmental Laboratory. Ifendogenous levelsofPFOS are found to be higher than the LLOQ or if the levels are found to be highly variable, the LLOQwillbeadjusted to accommodate this. The screening procedures and LLOQ determination will be recorded in a note to file and kept with the raw study data as welabeing reported in the final report. Recovery efficiency at the LLOQ will be determinedduringeach run using an extracted quality control sample. spiked at the LLOQ level. Ifthe quality control sample at the LLOQ level fails during a run, the LLOQ for that specific run would be considered 10 be the next lowest passing quality control sample and any samples below the new LLOQ must be reanalyzed "The upper limitofquantitation (ULOQ) was determined to be 250 ng/g in liver and 250 ng/mL in serum during the method validation as described above in section 11. Anextracted quality control sample spiked at the ULORQ will be used tomeasurerecovery efficiency at the ULOQ. Ifthe quality control sample at the 'ULOQ fails durinag run the ULOQ for that specific run wouldbeconsidered tobe: the next highest passing quality control sample and any samples above the new ULOQ must be reanalyzed. `Any sample wiantarhea greater than thatof the highest acceptable standard wil need to be diluted into the rangeofthe calibration curve. Ifsamples are diluted into the rangeofthe curve during analyses and enough sample remains, a post-run dilution validation will be performedtoverify sample values. To perform the 3M Environmental Laboratory 6 3M Environmental Laboratory Pose 198 Report EOI-1245 PFOS: A Reproduction Study with the Nother Bobwhite d(iiel.uttiownova1limdLatiaolni,quoontes sfoarmpfllueidwislalmbpelesseoprartawteod 1ingtroatmwaomroeupnretssenftoarttiivsesuseamples sdialmuptliensg)thtehesnadmiplluetewdituhsiandgdittwioonaplromcaedtuirxesp.rioTrhteofeixrsttrapcrtoicoend(usreeracoandsdiisntsg osefra), `ewxhtirlaecttihoens(emceotnhdanporloceexdturracetcaodnsdiisntgsoafdddiitliuotnianlgmtehteheaxntorlacstolwvietnht)s.olvent post- Ifthe values are not within 15%ofeach other additional testing will be required to determine which value is a correct representation ofthe sample concentration. 124 Continuing Calibration Verifications . bAenaunnaelxytzreadctaefdermiedvieervyel1s0tiannjdeacrtdio(ncso(nsttianrutiinnggacfatleirbrtahteiolnasveirnijfeicctaitoinono--ftCheCV) will calibration curve). The accuroafcthyeCCV must be within 20%. Data may only bcealsisbproatritoandciurnvaesphoointta).d iDyaptaassbiroanckCetCe'd sbyoonr8e oCrCmVorae0)faHileinsgaCnCsV'losvem)ust be reanalyzed. 125 Quality Control Samples Aminimumofthree (3) quality control samples (QC) will be prepared with cach set ofsamples. They wil consisofblank matrix spiked with three levels of anaLlyote.w TlehveelleavteltshearLeLOQ (10 ng/mL in serum and 10ng/g in liver), Midlevel: Highlevel: atttihmeesUtLheOLQL(O2Q5,times the LLOQ) 126 EQaCchPQeCrfiosremxpaenccteedCtroitsehroiwa an accuracyof80-120%ofexpected. A mirimum of2/3ofall QC must meet these criteria. Ifnot, the entire sample set must either be re-analyzed or re-extracted (as decided by the Study Director). 127 Use of Confirmatory Methods Confirmatory methods are typically not needed with LC/MS/MS analysis 12.8 PDFemOoSnssptecriafitciitoynwiollfbSepescuibfstiacnittiyated by chromatographic retention time, by the characteristic primary ion (495), and the characteristic product on (99) using LCMS/MS. 13. SUB-CONTRACTED ANALYSIS 13.1 AplelrfloirmdedanatdPtaicsesuTeieerxt2r,ac1t7i0o0nsElams Sdterteaeitl,edSuiinteth2i0s0,prMoitnonceoalpoalimse,nMdmNen5t54w14i.l be SM Environmental Laboratory 7 3M Environmental Laboratory Page 197 CHENG Report EO1-1245 PFOS: A Reproduction Study with the Nother Bobwhite: 13.2 All fluid and tissue analyses as detailed in this protocol amendment will be performed at 3M Environmental Laboratory, Building 2-3E-09, 935 Bush Avenue, St. Paul, MN 55106. 13.3 Pace Analytical Services, Inc. will follow all applicable 3M Environmental Laboratory Standard Operating Procedures and the following Pace Analytical In. Standard Operating Procedures PSS-Admin-01 Quality System PSS-Admin-04 Laboratory Facilities PSS-Admin-06 Training Record Files Maintained by QAU PSS-ARC-02 DispositionofArchive Materials PSS-ARC-04 Archiving Data, Specimen and Logbooks PSS-DC-05 Lab Notebooks, Logbooks, and Phone Logs PSS-DM-03 Statistical EvaluationofData PSS-OPS-01 Use and MaintenanceofUltra-Turrax T25 Homogenizer PSS-OPS-02 Raw Data PSS-OPS-03 Hazardous Waste Disposal PSS-OPS-04 Use and MaintenanceofN-Evap Analytical Evaporator PSS-OPS-05 UseofCompressed Gases in the Laboratory PSS-OPS-06 Use and Maintenance of NANOpure Il Water Purification System PSS-OPS-07 Cleaning Non-disposable Volumetric Pipettes PSS-OPS-08 Laboratory Calculations PSS-OPS-09 Deviations from Established Procedures PSS-OPS-10 Analytical Definitions PSS-OPS-11 Use and Calibrationof a Bottle-Top Dispenser PSS-OPS-13 Use and MaintenanceofLab Refigerators, Freezers, and Ovens PSS-OPS-14 Use and Calibrationof Syringes with a Hamilton Chaney Adapter PSS-OPS-15 Glassware Cleaning PSS-OPS-16 Use and Maintenanceofthe Fisher Scientific Isotemp Freezer PPSSSS--OOPPSS--1187 UUssee aanndd MMaaiinntteennaanncceeooffVUlotrrtaesxonSihcaWkeartser Baths (Soricators) PSS-OPS-24 Use and Maintenance ofa Desiccator PSS-OPS-31 Use and MaintenanceofShakers PSS-OPS-32 Operation, Maintenance and CalibrationofLaboratory PSS-OPS-37 Operation, Maintenance, and Calibrationof pH Meters and pH. Electrodes PSS-MC-01 PurchasingofLaboratory Supplies PSSMC-02 ReceiptofLaboratory Supplies PSS-MTR-01 CalibrationofCertified Weight Set PSS-MTR-02 CalibrationofCertifiedThermometers/Thermocouples PSS-MTR-05 Calibrationof EppendorfPipettes PSS-MTR-06 VerificationofCalibrationof theEppendorfPipettor PSS-MTR-07 New Equipment Qualification (IQ, PQ, 0Q) PSS-MTR-08 VerificationofCalibration and UseofAnalytical Balances 3M Environmental Laboratory 8 3M Environmental Laboratory Page 198 onrgL Report EO1-1245 PFOS: A Reproduction Study with the Nothern Bobwhite PSS-MTR-09 PSS-Safety-01 PSS-Safety-02 PSS-Safety-04 PSS-Safety-05 PSS-TS-01 UMosneiatnodriMnagiDnetveincaensceofFisher Scientific Electronic Temperature GUesneeraanldLMaabionrtaetnoarnyceSoaffeBtiyological Safety Cabinets UEsmeeragnedncMayiEnvtaecnuaantcieoonfPLraobcoedruarteosr.y Fume Hoods Training Procedures 13.4 pAernfaomremneddmaetnltabtoortahtoeripersoottohcoelrtwihlalnbtehewPriatcteenTiifeexrt2raocrti3oMnsEnavnidraonnamleysnetsalare Laboratory, respectively. . 14. STATISTICAL ANALYSIS SEtxaatimsptliceaslomfetthheodcsalwciulllatbieonlsimuisteeddtion tthhee acnaallcuylsaetsiwoinolbfemienacnlsuadnedd isntatnhdeaarndaldyetviicaatliopnhsa.se report. 15. Report foAlflionwalarlelpGorLtPwirlelqbueiprremeepnatrse,dabnydtwhiell3iMncElnuvdaierodnemscernitpatliLonaobofraaltlomrayt.erTihaelsiannadl rmeetphoodwrsitl used, including a narrative discussion witha tabular presentationofalanalysesresults 16. LOCATIONOFRAWDATA, RECORDS, AND FINAL REPORT `reWphoernt,tshteufdiynaclorrerpeosrptoinsdceonmcpel,estaemdp,lael roerciegiipntalanpdapterracdkaitnag,, isnacmlpuldienpgrperpoatroactoilo,n,pahansde acnoarlryetsicpaolnddiatnag,twrialilnbineg rreetcaoirndesd, icnaltihberaatricohnivreescoorfds3,MinEsntvriurmoennmtemnatianlteLanbaonrcaetolroyg.s, Asltlandard `Toipeerra2tionrgtphreoc3eMduErnevsi,reoqnumiepnmteanltLparboocreadtuorreys,,aasnadppmleictahboled.s wilberetained either at Pace 17. Specmen Rerenmion `anSaalmyptliecsaolfstthanedatresdtsswuilblsbtaenmcaei(natnayinleedftionvetrhecolnatbroorlatsoerrya ofroairveprerainoddosfatmiplmees)asansdpetchiefied by r`seagmupllaetsiownilolrnaostlboengmaaisntthaeinqeudalmiotyroeftthhaenp1r0epyaeraartsioafntearfftohredsefefveacltuiavteiodna.teoHfowtehveefri,na test rauplpel(yiftaopsplaimcpalbeler)e.te3ntMionEntviimreosnmental Laboratory Standard Operating Procedures also 18. REFERENCES 181 Wildlife Intemational, Lid. Protocol with the Northern Bobwhite" No. 434-108, "PFOS: A Reproduction Study 18.2 MCeentthroedAsna"lEyxttircaacltiLoanboorfaPtootraiesss,iIunmc.PeSrtfulduyor0o2o3c-a0t5a4n,es"uVlafloindaatteioonroOftAhnearlytical FElleucotrroocshpermyi/cMaalsCsoSmppeocturnodmsetFrryo"manSder"uExmtrfaorctAinoanloyfsiPsoUtsaisnsgiuHmPLC 3M Environmental Laboratory 9 3M Environmental Laboratory Page 199 Gots Report E01-1245 PFOS: A Reproduction Study with the Nothern Bobwhite PAnearlfylsuiosroUoscitnangcHsuPlLfoCnaEtleeocrtrOotshperrayF/lMuaosrsocShpeemcitcraolmeCtormypionuQnudaislFSreormumLiavnedr Lfiovrer." 18.3 3M the Study U2723, Wildlife Determinationof PFOS Study 454C-110, in Avian Diet." "Analytical Method Verification for 19. PROTOCOL AMENDMENATNSD DEVIATIONS PSltaundnyeDdicrehcatnogresatnodtthhee SprpootnoscoorlRweiplrebseenitnattihvee.forAmmoefnwdrmiettnetnsawmiellnbdemceonntssidseigrneeddabsyptahret offotrhmoefpwrroitotctoelnadnevdiawtililonbse, astitgancehdedby10thteheStfiundayl Dpriorteocctoolr. aAnndySoptohnesrorchRaengpersewsiellntbaetnainvthede filed with the raw data." Allchangesto the protocol wilbeindicated in the fina report 3M Environmental Laboratory 10 3M Environmental Laboratory Page 200 RSG PALIDISY FAK 410832 0832 WILDLIFE RRGSSATRgadduction Study with the NottsBobwhite -- WILDLIFE INTERNATIONAL, LTD ] PROTECTNO: E45o6v-12068,5 AMENDMENT TO STUDY PROTOCOL STUDY TITLE: PFOS: A REPRODUCTION STUDY WITH THE NORTHERN BOBWHITE PROTOCOL NO.: 454/120700/QR/SUB4S4 SPONSOR: 3M Corporation AMENDMENNOT. 10 PROJECTNO.: 454108, E0L-1245 EFFECTIVE DATE: January 30, 2002 AMENDMENT: This amendment inclades an stachmen: (Aachen A) which provides updated formation necessary `complete the analytical phaseofthis study. This attachment A replaces `the attachment A in amendment 9. ReasoN Thc upd amehnedmdenntquaalaictyhmbjeenctivAesw,sanudpd changeintchleudveeradainfderseenst etxrtarcacttiioonn acnadtainoanlyialmeth,to SeaFn. GulagPh.erh, Sdtudy Diocior Bi don E TET T Laboratory Management B odd = Harold O. Johnson, Principal Analytical Investigator pi>lloK Regen, 30)Evomamentl Caborstor, Lm a ---- RofefgcRd~obJii deaes)eSpoS njJorEs < RepreseCaUAtative tele (3 lo2 Date. Bae Cll Dats 2 174 3M Environmental Laboratory 10f9 GoxiPaigel201 Report EOI-1245 WILDLIFE INTERNATIONAL, LTD PFOS: A Reproduction Study with the Nothern Bobwhite PROJECT NO: 454-108, E0L-1245 AMENDMENT TO STUDY PROTOCOL STUDY TITLE: PFOS: A REPRODUCTION STUDY WITH THE NORTHERN BOBWHITE PROTOCOL NO: 454/120700/QR/SUB4S4 AMENDMENT NO: 10 SPONSOR: 3M Corporation PROJECT NO: 454-108, E0I-1245 EFFECTIVE DATE: January 30,2002 AMENDMENT: -- -- t"oThcisomapmleentdemtehnetanianlcyltuidceasl apnhaastesocfhtmheinstst(uAdtyt.acThhmiesntatAi)acwhhmiencth AprroevpildaecseusptdhaetaedttiancfohrmmeantAitoninnecessary amendment 9. REASON: "uTphdeataemethnedmdaetnatqu9aaltittyacohbmjeecnttivAesw,aasnduptdoatcehdantgoeitnchluIdveeardainfdfesreernateexxttrraaccttiioonn laoncdataionna.lytical method, to Sean? GallagherS,mdy Diesor Dae LJaobaonrnaBt.orByeaMvaenras,geWmiednitfe International, LTD, Fiarold0. Johnson, Principal Analytical Investigator Date -- Bae LWaibloiraamtoKr.y MRaenaaggeenm,3enMtEnvironmentalLaboratory, [TI Rochelle R_ Rabideau, Sponsor's Representative Date -- 10f9 3M Environmental Laboratory Page 202 COLABE Report E01-1245 PFOS: A Reproduction Study with the Nothern Bobwhite Protocol Amendment # 10 - Attachment A Wildlife International Ltd. # 454-108 STUDY TITLE PFOS: A Reproduction Study with the Northern Bobwhite: 3M EnvirSoPnmOenNtSalORLaboratory Building 2-3E-09 PO Box 33331 St. Paul, MN 55133-3331 PERFORM3IMNEGnvAirNoAnLmeYnTtIalCLAaLboLraAtBorOyRATORY Building 2-38-09 St. PauPlO, MBNox 5353133331-3331 LAB3OMREAnTviOrRonYmePntRaOlJLEabCorTatIorDyE:NTE0I1F-I1C2A45TION `Wildlife Intemational Ltd.: 454-108 3M Environmental Laboratory 20f9 3M Environmental Laboratory Page 203 60126 Report EO1-1245 PFOS: A Reproduction Study with the Nother Bobwhite: Sponsor Study Identification PFOS: A Reproduction Study with the Northern Bobwhite 3M Environmental Laboratory Building 2-3E-09 PO Box 33331 St. Paul, MN 55133-3331 Principal Analytical Investigator SAtNuAdLyYTLIoCcAaLtTiEoSnTsING LABORATORY Harold Johnson 3M Environmental Laboratory Building 2-38-09 PO Box 33331 St. Paul, MN 551333331 651-778-6479 3M Environmental Laboratory Building 2-3E-09 935 Bush Ave. PO Box 33331 St.Paul, MN 55133-3331 Proposed Study Timetable ANALYTICAL EXPERIMENTAL STARTDATE ANALYTICAL EXPERIMENTAL TERMINATION Dare 01/31/02 03/31/02 aM Environmental Laboratory 3019 3M Environmental Laboratory Page 204 CHALR? Report EO1-1245 PFOS: A Reproduction Study with the Nothern Bobwhite 1. PURPOSE/OBJECTIVES In the in-life phaseofthis study, Norther Bobwhite quail were regularly exposed to t`oneovmainlaulatdeostehseocfffpeecrtfsloufordoioecttaarnyeesuxlpfoosnuartee,uppoontaasdsuilutms osavletr(aPFfiOvSe)moinntthheiprerfieoedd. inTohreder purposeofthis analytical phaseof the study is to quantify PFOS in the liver and sera of both control and dosed Norther Bobwhite quail. The analytical information may be used to compare with toxicologicalhistopathological results from the in-life portionofthis bird study. The in-life portionofthe study was completed at Wildlife Intemational, Lid. in study 454-108. 2. REGULATORY COMPLIANCE "This analytical phase study will be conducted in accordance with the United States. Environmental Protection Agency Good Laboratory Practice Standards, 40 CFR 792. 3. QuALTY ASSURANCE The 3M Environmental Laboratory Quality Assurance Unit will audit analytical phase study conduct, data, and the final report to determine compliance with Good Laboratory Practice Standards and with 3M Environmental Laboratory Standard Operating RPreopcreedsuernetsa.tiTvheeanQduatlhietyStAusdsyurDainreccetoUrnit will report all findings to the Sponsor 4. REFERENCE SUBSTANCE 4.1 Physical Description- Perfluorooctanesulfonate, potassium salt from Lot 217 is a white crystalline material. 4.2 Purity and Stability--The purityof Lot #217 has been determined to be 86.9%. PFOS has been shown to be stable in methanol solutions for up to 6 months at ambient ItnesmtpreurmaetnutreDeintetchteivoanliLdiamtiitonDerteeprormtinfartoimonstfuodryLPCS/0M13S,a"nMdeGthCo/dMVSalAindaaltyisoinsoanfdFC807 Compounds in Methanol." Refer to the Certificateof Analysis (stored in the Test, Control, and Reference database in the 3M Environmental Laboratory) for further purity and stability information. 4.3 Storage Conditions--Th test article may be stored at ambient temperature or lower. 4.4 Reserve Samples--A reserve sample of PFOS from Lot #217 has been frozen at 20C 5C or colder and is kept in the 3M Environmental Laboratory. 4.5 CAS Number--The CAS number is: 2795-39-3 4.6 Molecular Weight--The molecular weight is 538 (CiFi:SOK). Note: The molecular weightof the anion (PFOS) is 499 (CsF11805). 3M Environmental Laboratory 40f9 3M Environmental Laboratory ory GO11IRE Report E01-1245 PFOS: A Reproduction Study with the Nother Bobwhite: 5. CowtRoL MaTRices `Typesofcontrol matrices and their source, physical description, storage requirements, and traceability numbers will be recorded in the raw data and included in the final report, 6. TestSystem The test system is the Northern Bobwhite quail. The test system was dosed (at Wildlife Intemational Ltd. during the in-life phaseofthis study) with food prepared by mixing PFOS with feed in a large mixer in appropriate portions to achieve the nominal 10 ppm level. A food dose verification validation study was performed by Wildlife Intemational under study number 454C-110 (3M #12723), "Analytical Method Verification for the DeterminationofPFOS in Avian Diet." Wildlife Intemational verified the actual dosed food for this study and the results will be contained in the in-life Wildlife phase report Norther Bobwhite serum and liver samples were collected by Wildlife International, Lid. and sent to 3M Environmental Laboratory for analysis. 6.1 Justification of the Selection of the Test System Northern Bobwhite quail represent wild bird populations and they are an EPA recommended species because they do well in a laboratory environment. The use ofthese matrices for the study is to determine ifPFOS was deposited in the liver and serumof mating adults that were fed POS. Furthermore, analysisof these matrices from hatchlings will determineifPFOS was passed on to mating adults" offspring. 7. SOURCE OF SUPPLYFOR THETEST SYSTEM The birds were obtained by Wildlife Intemational, Lid. See their protocol (study 454108) for information about bird origination location. 8. SPECIMEN AND SAMPLE RECEIPT AND MAINTENANCE Sera and liver were collected by Wildlife Intemational, Ld. and shipped to the 3M Environmental Laboratory for analysis. The 21-weck endpoint sera and liver samples received by the 3M Environmental Laboratory were from the following birds: 40 control adult birds (20 mating pairs) that ingested food containing 0 ppm PFOS 40 experimental adult birds (20 mating pairs) that ingested food containing 10 ppm PROS 20 first generation offspring birds that ingested food containing 0 ppm PFOS `Samples will be maintained frozen except when removed for extraction and analysis as described in the method. The samples will be kept isolated from the test substance during storage. 3M Environmental Laboratory 5019 3M Environmental Laboratory Page 206 GNIANG Report EO1-1245 PFOS: A Reproduction Study with the Nothern Bobwhite: 8. SUB-CONTRACTED ANALYSIS All sera and liver extractions and analyses as detailed in this protocol amendment will be performed at 3M Environmental Laboratory, Building 2-3E-09, 935 Bush Avenue, St Paul, MN 55106.55414. 10. EXPERIMENTAL CONDUCT The 21-week endpoint liver and sera samples received (see section 8) will be extracted and analyzed for PFOS quantities using the methods listed below. 10.1 Preparatory and Analytical Method 10.1.1 ETS-8-231 Solid Phase Extraction and Analysis of Fluorochemical Compounds from Biological Matrices 11. DATA QUALITY OBJECTIVES DURING SAMPLE ANALYSIS Method validation for PFOS quantitation in rat and mouse sera and liver is in process in the 3M Environmental Laboratory study EO1-1277. Limited Norther Bobwhite control matrix preclude a cross validation, however, the EO1-1277 study methods will be applied to sample analysesof Northern Bobwhite sera and liver samples using the following performance based criteria: 11.1 SSyysstteemmsuSituaibtialbitiyliwitlyl be determined prior to the start and at the completion of each analytical run. Prior to the calibration curve and after the last sampleof the run three (3) mid-level unextracted calibration standards will be analyzed. The peak area precision and retention time precision wilbe monitored at the beginning and the endof the run separately. The peak area precision must be equal to or less than 2.0% RSD, the precisionofthe retention time must be cqual 10 or less than 2.5% RSD. If any itemof the system suitability fails, system msaaimnptleesnaanncde tmhuesstybsetecmosmupilteabtielditpyrimoursttopraussnnbeifnaogressetacrotnidngsetthoefscaylsibtreatmiosnu.itability 11.2 Absolute Recovery "The absolute recovoeftrhye method will be evaluated one time in both liver and smeirnuim.muFmoorfetawcoh mlaetverlisxoafmPiFOnSimuthmaotfbtrraicpkleitcattheesraamnpgleeosfawnilallybteefdorettiefriemdinateda for the samples (i.c., at near the highest level determined in samples and at ~150% of LOQ for samples). The samples will be extracted through the method, and analyzed by comparison with an extemal calibrationofnon-extracted standards. "wiTlhle ncoonns-iesxttroacfatemdisntianmduamrodfcnaliinbera(t9i)onlecvuerlvs,esinwcilludbiengpraempaertehdanionlmebltahnakn.olT,haend k best appropriate regression will be used to describe this curve (for best accuracy at all levoeftlhse standards). 3M Environmental Laboratory 60f9 3M Environmental Laboratory Page 207 onLLs Report EO1-1245 PFOS: A Reproduction Study with the Nothern Bobwhite The accuracy (% absolute recovery) and precision (%CV of the recoveries) will be determined at each level using the external calibration curve for quantitation. "There is no control limit for accuracy; precision must be better than 20% at each level. Shoulda level (high or low) not meet this precision, the method will only be considered valid for levels between the two valid levels. Additional testing may be conducted at other levels to extend the acceptable rangeof the method. 11.3 Calibration A calibration curve should be prepared in each matrix by spiking the matrix with known concentrations of the analyte and surrogate. A calibration curve should consistofat least nine standard curve points covering the expected range, including the LLOQ. The simplest model that adequately describes the concentration-response relationship should be used. The accuracyofeach level will be verified. Any level outside 75% - 125%ofnominal must be deactivated, and regression re-calculated, except the LLOQ which must be within 30% of nominal. All levels must show a response greater than twice thatofthe matrix blank. A maximum of four (4) levels may be deactivated in any one set, or the set will be re-analyzed. 11.4 Limits of Quantitation (LOQ) "The lower limitofquantitation (LLOQ) willbedefined in liver (ng/g) and in Serum (ng/mL) as the lowest acceptable extracted calibration curve point that is within the 70% -130% nominal criteria and with an analyte peak area at leas2t. times the blank. The LLOQ determination will be recorded in a notteo file and Kept with the raw study data as well as being reported in the final report, "The upper limitofquantitation (ULOQ) will be defined in liver and in serum as the highest acceptable extracted curve point that is within the 75%- 125% nominal criteria and includes applicable dilution factor corrections. 11.5 Quality Control Samples A minimumofone sct of nine (9) quality control samples (QC) will be prepared one time in each matrix. They will consist of three levofeanallytse, each in triplicate. The quality control samples should be prepared in each matrix by spiking the conirol matrix with known concentrationsof the analyte. The levels bracket the sample concentrations determined in the study: + Low level: equivalent to an analyte concentration nar orbelow the lowest level ofanalyte determined in samples + Mid-level: equivalent to an analyte concentration near the middie of the sample range High level: equivalent to an analyte concentration near or above the highest level of analyte determined in samples 11.6 Quality Control Sample Acceptable Recoveries 3M Environmental Laboratory 7of9 3M Environmental Laboratory Page 208 Gnaiss Report E01-1245 PFOS: A Reproduction Study with the Nothern Bobwhite Each QC is expected to show an accuracyof75-125% of expected. A minimum of2/3 (6 outof9)of all QC must meet these criteria and aminimumofone half (2 out of 3)ofthe QC at each level must meet these criteria. Ifnot, the entire sample set must either be re-analyzed or re-extracted (as decided by the Principal Analytical Investigator). 11.7 Matrix Spikes Two matrix spikes wil be extracted with each sample preparation. The matrix spikes will consistofone fortification level in the rangeofthe samples, extracted in duplicate. If the marx spike and/or matrix spike duplicate fall outsideof25% accuracy, the curve, CV's, matrix spikes and samples will be reanalyzed. 11.8 OBnleanckonstrol matrix and one water method blank will be extracted with each sample preparation. 11.9 Continuing Calibration Verification Two extracted standards (continuing calibration verification ~ CCV) will be analyzed after every 10 injections (starting after the last injection ofthe calibration curve). The accuracyof the CCV must be within 25%. Data may only be reportedif bracketed by passing CCV's (ora CCV and the same level calibration curve point). Data bracketed by one or more failing CCV must be reanalyzed. 11.10 Use of Confirmatory Methods Confirmatory methods are typically not needed with LC/MS/MS analysis. 11.11 Demonstrationof Specificity PFOS specificity will be substantiated by chromatographic retention time, by the. characteristic primary ion (499), and the characteristic product ion (99) using. LC/MS/MS. 12. SUB-CONTRACTEDANALYSIS 121 All fluid and tissue extractions and analyses as detailed in this protocol amendment will be performed at the 3M Environmental Laboratory, Building 23E.09, 935 Bush Avenue, St. Paul, MN 55106. 122 An amendment to the protocol will be writenifextractions andanalysesare performed at laboratories other than the 3M Environmental Laboratory. 13. STATISTICAL ANALYSIS Statistical methods will be limited to the calculation of means and standard deviations. Examplesof the calculations used in the analyses will be included in the analytical phase report 3M Environmental Laboratory 8of9 3M Environmental Laboratory. Pe Report E01-1245 PFOS: A Reproduction Study with the Nothern Bobwhite 14. ReporT A final report will be prepared by the 3M Environmental Laboratory. The final report will follow all GLP requirements, and will include a description of all materials and methods used, including a narrative discussion witha tabular presentationofall analyses result. 15. LOCATIONOFRaw DATA, RECORDS, AND FINAL REPORT `When the final report is completed, all original paper data, including protocol, phase report, study correspondence, sample receipt and tracking, sample preparation, and analytical data, will be retained in the archivesof3M Environmental Laboratory. All comesponding training records, calibration records, instrument maintenance logs, standard operating procedures, equipment procedures, and methods will be retained at the 3M Environmental Laboratory, asapplicable. 16. SPECIMEN RETENTION `Samplesofthe test substance (any leftover control sera or liver and samples) and the analytical standards will be maintained in the laboratory fora period of time as specified by regulation or as long as the qualityofthe preparation affords evaluation. However, samples will not be maintained more than 10 years after the effective dateofthe fina test rule (if applicable). 3M Environmental Laboratory Standard Operating Procedures also apply to sample retention times. 17. RereReNCES 18.1 Wildiife Intemational, Lid. Protocol with the Northern Bobwhite". No. 454-108, "PFOS: A Reproduction Study 18.2 3M Study U2723, Wildlife Study 454C-110, "Analytical Method Verification for the Determination of PFOS in Avian Diet." 18. PROTOCOL AMENDMENTS AND DEVIATIONS Planned changes 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 ofthe protocol and wil be attached to the final protocol. Any other changes will be in the form of written deviations, signed by the Study Director and Sponsor Representative and filed with the raw data. All changesto the protocol will be indicated in the final report. 3M Environmental Laboratory 90f9 3M Environmental Laboratory. Page 210 Gna Report EO1-1245 PFOS: A Reproduction Study with the Nothern Bobwhite WILDLIFE INTERNATIONAL, LTD PROJECTNO.: 454-108, EO1-1245 AMENDMENT TO STUDY PROTOCOL STUDY TITLE: PFOS: A REPRODUCTIONSTUDYWITHTHENORTHERN BOBWHITE PROTOCOLNO.: 454/120700/QRISUB4S4 AMENDMENT NO: 11 SPONSOR: 3M Corporation PROJECNOT.: 454-108, 01-1245 EFFECTIVE DATE: March 27, 2002 AMENDMENT. Swion 11% Semienced Change:rTetheenptieoanktairmeae mpruesctisbieocnqmuuasltoboereqlueaslst1haonr2l.e5s%s tRhSaDn.2.0% RSD, the precisionofthe "To:rTehteenpteiaonkatrimeeampruesctibsieoenqmuuasltbooe cqlueaslstthoaon2l.e5ss%tRhaSnD.5.0% RSD, the precisionofthe REASON:aIrnefaocromuantitoinscreocmeimvoend ftorUomVoduertiencsttorrusmpeenctifsiucpaptliioenrs.inMdaicsastsesptehact at2r.o0%mewRthSieDcfrhoswrte,h.e ausnebdfeorstuhpiprsetsusdeyd,bayroethmeorrsepseencsiietsipvree,sienagteinnetrhalesaanmdptleefmaacttrtihxataanndamloybtielieonpihzaasteion cmlaekanelianreesasraeprrotdhuecidbeitleirtmyimnoinrgefcahcatlorlse,ngbiuntg<t2.ma0i%nitsaisn.lSaambpitlaegagnrdesssoiuvercfeor mass spec. The application chemists at Micromass, the instrament vendor, suggesta value of <5.0%. AMENDMENT: Section 113, Sentenc5e Change: rAencaylcluelvaetleodu,tesxicdeep7t5th%e-L1L2O5Q%owfhincohmimnuasltvbaeluweitmhuisnt3b0e%doefactniovmaitneadl,. and regression "To: iInncsluodmeedciansteshtlhienclaorwersatngceurtoveinpfolnute,ncaeltthheoulgohweourtesniddeotfhteh3e0cu%rvne.omiOnnallyroenqeuiproetme,ntt,hemlaoywebse.t lequvaenltiptoaitnito,n.caTnhbeelliemfittoifnqfuoartnhtiitspatuiropnosweil.Iltbewitlhlennootbxepuoisnetdotnodtehteecrumrivneetmehctliinmgittohef30% nominal requirement. REASON:cTalhiebrraetmioonvaplooifntthoedleovwieasttecbayligbrreaattieorntcuhravne3p0o%i.ntReimnsoovamlecoafstehsicspaouisnets itnhtenuenxctaluosweesst ctahleinberxatticoanlpiobirnattisoinnpsoionmtetcoadseevsicatoeulbdygcroenattienrutehuantnil30th%e.reThariesvceornytifneuwedpodienltestion of 3M Environmental Laboratory Page 103 Coles Report EO01-1245 PFOS: A Reproduction Study with the Nothern Bobwhite 3r0em%aiisniunsgeidntthoiencfulruvcen,ceTthheeilnocwleusrieonndoofftchuercvuerpvoeianntwdiitsnhoatduesvieadtfioorndgerteeatremritnahtainon ofthe LOQ. AMENDMENT: Section 11.3,thelstsentence, Change: Areamnaalxyizemdu.moffour (4) levels may be deactivated in any one set, or the set will be To: The calibration curve will contain a minimumofsix (6) calibration curve points. REASON:"TLnhemloasrtgocansuesmtbhercoaflipboriaattisoinntcuhrevceusrcvoentiasuinsgerdeattoesrctlheacnt nirnaeng(e9r)ecparleisbernattaiotnivpeotinotts.he csaalmipblreast.ioLnirmaintgiengnotthererpermeosveanltaotfiovneolfythfoeursa(m4)plpeosints in some cases could cause a AMENDMENT: Section 113 `Add: The coefficientofdetermination or the calibration curve should be > 0.985. REASON: Nro ceoeffmipcriioeontotvfodperetienrtdimnignoafttiohneifsinianlccluodpeyd.in the protocol. It was inadvertently 3M Environmental Laboratory Page2af3 Page 212 Canann ReFpQ 1o245s17 Fax 410 22 0632 wruovrre FRSARggipduction Study with theNope Bobwhite aoe 1:25 RSD IN TEDH 2-28-01 + ~a1GG2SLS ro.505 peor i! j VKDLIFE INTERNATIONAL, LTD. " Dame 8 Sel .Gallagher, rRomCTNO 454108, J eres SELLE : Major 'B. Ieenational, LTD % g eR O Pa AR eg DE : by Ri: Sez. ! f "Williaa K_ Reagea, IM Environments) Laboratory, Date: Laboratory Manageme SES nd Sprasoy's Represcatative 7 . o i iy ie ' 3 i 8 3M Environmental Laboratory i : Psat | Page 213 COLL Report E01-1245 "WILDLIFE INTERNATIONAL, LTD PFOS: A Reproduction Study with the Nother Bobwhite: PROJECT NO.: 454-108, E01-1245 Sw Ghge SDR Joann B. Beavers, Wildlife Intemational, LTD Laboratory Johnson rincipalAnalytical Trvesigaior [oE mK.Reage in, 12EnvironmentalLabora, Laboratory Management "HohelRiK e Robideau, Sponsoy's Representative bw Date. Tw4.2-02 aie Oy --o>-< > 3MEnvironmental Laboratory Page 303 Page214 GAS? Report EO01-1245 WILDLIFE INTERNATIONAL, LTD PFOS: A Reproduction Study with the Nothern Bobwhite PROJECT NO: 454-108, EO1-1265 AMENDMENT TO STUDY PROTOCOL STUDY TITLE: PFOS: A REPRODUCTION STUDY WITH THE NORTHERN BOBWHITE PROTOCOL NO: 454/120700/QRISUB4S4 AMENDMENT NO: 12 `SPONSOR: 3M Corporation PROJECT NO: 454-108, EOI-1245 EFFECTIVE DATE: August 16,2002 AMENDMENT: Cthheansgteu:dyTishEeO13-M12E4n5v.ironmental Laboratory Project Identification Number (LIMS) to be included with "thTo:sTtuhdeymiasiEnD13-1M24E5n.viArdodnimteinotnaall LtraabcokriantgornyumPbreojresctasIsdoecnitaitfiecdawtiiotnhNtuimsbsetrud(yaLrIeMSU)27t2o3b,eEiOnIc-l1u3de4d2,wiatnhd EOI-1365. REASON: BwoebrwehaistseigsnaemdpaleLsIwMeSrtrraecckeiinvgednuimnbtehre. 3NMotEnavwiarroenmoefntthaelsactospeeveorfahl itsimoer paoniyntso.theUrppoarntiacrurliavarl,stsuadmyples paaescrsthiaginonefidntghteotsotehtipshreossjteeucsdtaysmippnlecerltsua,dietnihtnehgyetwfoeortlhelioscwhisnteguc:dkyeUd2w7ei2rne3t,oandEaiOfl1fy-ez1ree2dn4t5u3,ndMEeOrL1o-In1Me3S4m2pa,rioanjnescdttusE.dOyIL-n1Iu3Mm6Sb5e.prrSoEjaOemIcp-tl1ne2us4m5ib.ners AMENDMENT: Change: Harold Johnson is the Principal Analytical Investigator. To: Lisa A. Stevenson is the Principal Analytical Investigator. REASON: To change role of the Principal Analytical Investigator 3M Environmental Laboratory Page 102 Page 215 CLLRS Report E01-1245 PFOS: A Reproduction Study with the Nothern Bobwhite WILDLIFE INTERNATIONAL, LTD Ase Sean. Galla ghber , S ady Biecor I Toa Beatd ers, WI ie mermaonl,n LTD, Livoratory Management utheveSnsoen,ePnomp Amy vesigtor aKDAue gen It M Evonmental ahora. Laboratory Management oTs =opa[oiA n e,d Sy rstlD Rereenais PROJECT NO.: 454-108, E01-1245 Baeqfx3s5/0d aBoe bder Broe tha Bieciatoiloa Ba s/oJu 3M Environmental Laboratory Fapezor2 OLE Page 216 Report E01-1245 PFOS: A Reproduction Study with the Nothern Bobwhite: WILDLIFE INTERNATIONAL, LTD PROJECTNO.: 454-108, EO01-1245 AMENDMENTTO STUDY PROTOCOL STUDY TITLE: PFOS: A REPRODUCTION STUDY WITH THE NORTHERN BOBWHITE PROTOCOL NO: 454/120700/QRISUBAS4 SPONSOR: 3M Corporation EFFECTIVE DATE: September 27, 2002 AMENDMENTNO: 13, PROJECT NO: 454-108, EOI-1245 FT Change: Rochelle R. Robidean i the Sponsor Representative Tor Susan A. Bach i the Sponsor Representative. REASON: "To change ole ofthe Sponsor Representative ScaLn P.aGalelaghetr,.Study Dlirelct s. Diehoy r Tom BeerR , WHA Terma LTD. Laboratory Management OsanA. AS.teveSnlseonr,tPrreinncipal Analytical nvesigator p William K.r ReZagen, 3M --Environmental Laboratory, Labgrstory Management Tre o\\er Datoed/azloa aS hosbs ate Sudin A. Beach, Sponsor's Representa2tive Taitje3a7 [eo 3M Environmental Laboratory Page tof Page 217 a1Lan Report EOI-1245 3M Confidential PFOS: A Reproduction Study with the Nothern Bobwhite Record of Deviation ESt0uld-y124P5roject No. D(eCvhieactkioonntey)pe SOP Protocol Document number ETS4-40 I__Identification OMethod 0 Equipment Procedure [] Other: Dates)ofoccurence 23/1121/0022,, 22711301202,,12310520,22702, 28002, 21102, Ti Description ERTeSq-u4i-r4e0dIpnreomcaelducrhei/nporofcceussso;dy W"iTthhiinntetrhnealIac.haTiheofcfuosrtmsohdoyulfodrbmetlobeeduwitshtostreuadcydkdaatla omr oothverereolmefvasenatmnpslatemsple documents. AInctteurmaall cphraoicnseodfucrues/tpordoicseswse:re not used 1 rac samples nd sample extracts. Since recipt samples were Stored in freezer FI. After homogenizing lve and alquaisenrgsamples werestored in Freeze FS Uniextractiondate. Samples remained frozen uniextraction. Devition to SOP writen. suchas amTeiln,dmAecnttissounesd,TSaOkPerenvision, ic Recorded by, Authorizebdy Shy Duech J ot . ufeuske Sn A" Date 4 Foz Datg . ~~ Caged SyDDeceovriaiPorneNLoo.dFe TST R|yoFRED reasTiller asdor booby Yorgounk: grtrl ky tachment A DocumeniatoEnoTfSD4ev8i2ations foe/o- Pagesof | 3M Environmental Laboratory Page 218 6033 Report E0I-1245 PFOS: A Reproduction Study with the Nothern Bobwhite Record of Deviation T. Identification Study/ Project No.EO1-1245 Deviation Type (Check one) (SOP Method 0 Equipment Procedure Protocol 1) Other: Document Number-Protocol #: 454/120700/QRISUB454, amendment #: 10 2Da1t1e/(0s2),of21o2c1c0ur2r,en2c1e3:/02:,1/321//1042/,022/,1/20/21,8/20/22,/0421,12/1072/,02, aneo2 Ii. Description: ? Required mid-level uPnreoxcteradcutreed/cparlciebsrast:ioSnescttainodnar11d.1w,ilSlybseteanmalSyuziteadb.ility, second sentence says that a OAcntuoaclcaPsrioocnetdhurroeulgphrotcheesdsr:oppingofcalibration points, the system suitability samples were on uthneexhtirgahcteenddcoufrvtehepcouirntvse tohreiorutasriedaectohuentwsorwkeirnegatratnhgeehoifgthheencduorfvte.heAclatlhiobruagthiotnhecsuervweerraenge, Tthheey ddaotapafsrsomth2e/s7y/s02tewmassuijtuasbtialibtoyvcerittheericaaalinbdratthieosne rdaantgae.pacTkheetsewidlaltbaewcilolnsaildseorbeed caocncespitdaebrleed. acceptable (such as Til. Actions Taken: amendment issued, SOP revision, etc) Issuingofthis protocol deviation. wl 4 Date {Ah IV. Impact on Stu/Prdojeyct Gita a"Tnhderweewriellwibtehinno tihmepaacccteoptnatbhleesptruedciysaisonthreanignjeescetivoenns tihnovuoglvheadbwoevreetuheseedxtfroarctseydstcealmisburiattaiboinlity curve Authorized By (Stvuay Direftor// ProjeScet Lead) Date Grez 3H EFmovirRrmaErEmTeESna4l3L0abTora R deen o(ogoRby Sy DiorDovPioapteioLenndNo.hcc_n_3_or_pr_o) 3M Environmental Laboratory Page 219 GOAw IT Report E0I-1245 PFOS: A Reproduction Study with the Nothern Bobwhite Record of Deviation I. Identification Study/ Project No.EO1-1245 D(eCvhieactkioonnTe)ype 0VsProoptocol 0 MOetthhero:d 0) Equipment Procedure Document NumberETS-8-231.1 2Da1t1e/(0s2),o2f/o1c2c/u0r2r,en2c/1e3:/10/23,1/20/21,4/20/21,/022/,182//022/,024,1121//70/202,, : anei02 I Description: ; Required Procedure/process: Section 12.1, Calculations: Ifother calculations are used then those listed, they will be documented in the raw data. FSAocaotnuudgamaliEtPOyrIom1c3o4e2d|-um3rL6eo0/1fp,rePorecepepsosmhsoc:stednetsec.ptiTohns""hCoimrovgeenPaotie c-o1n0sptpoifn5IveI"n,"waussr4ep5pme.dwbalyers.pikSiinngce0I0..ou0t20f0016m6 iHnodmopgreepnaarteedsuswiengdS1PEr,etphecfarcah)cexrsaceie1d 2s.a0nmd.ro,fM1e1OHo5,ft0hTeH5ER0E1F5O)RoEf. ver 5spiked. Aer this homogenates spiked (20) (L.00gPFOS mk.McOH)(1Om 000)(1000n/1.=0u2g0)n PROS onoSPE column {ST2i0nh0egcPatFhOeiSs't/shtea2rne0dfamorrd hiMerOuencHiotrsd=bede1n30gng1m0PepFsbOs,Sehmdeln.cdoMme1cOetHpu.oniidsoarneth0echrnoma4tognPraLpoEhfiMcQEsyOsSHt). Simry the following semenisve. (20mg PROS: 11.0mL homogenat(e1 Om homogente 0.1 liver) = 200ng PROSg iver Tcihrsormlaatoongsahpihpehreosuwlst htah iiomr/olf2.0 sd coe" he unis when clean the fa est n ng/g fom he `SSEanOoIl-e1C3al4a2l-a3t0i3o4n8:,Opapim. 200 Mlver" HChormoomgieongeadphwiichs5.l0miL 1t0e76,nandgILre(pone0odfva%ilsuesn2r08w4eingghe,d,Ssiinnccee hev1er03d3i3ymiotfs1.o00laeswesighutomfieesredd 5f)or he cnt SPE procedure, th clclation 13 lows (1076ng PROS! mL MOH) (ml MeOH 0.1053 g vr) = 208. ng PROS g liver SMSamiEpmCleHeCEsaOh1le-t1dSo4n3nt-:36I0m17,iveQrC@h1o0pgghe1r" This QC samples prepared by adding 4ul.of 20013 (5.02005PEOS mi. (44) (5.02ugmL) (1mL/100L) (1000mgig) (0.1liv=e20r05)08 PFOS 5 ivr PUsRiOnSg t/hgevaelr,ethcelerleatcioonv,ste(hre7a1yna$ly2t0ic0alEe1s0t0)of 13. S1395.n4%g/mL calculates 0271 Sng ve. Since the spike vel is 200.805 3M EFnovrirmonEmTeSn.t4a:l8L0aboratory 3M Environmental Laboratory. Deviation No. _1 (asp by SyDirectoror PcLenda i cdof dyor REA, Report EO1-1245 PFOS: A Reproduction Study with the Nother Bobwhite Record of Deviation Ti, Actions Taken: Issuing ofthis SOP devia(tsiuocnhaansdaamenontdeme10ntfilies.sued, SOP revision, ei.) Recorded By ) he (0 Ao <S IV. Impact on St/uPrdojeyct "There will be no impact on the study as this is for clarification purposes. Date G15 op Authorized ByYo (Study Director /Project Lead) Sedole ResAL <S Shay Ocho: 2 . Men nln Date {aor Uburckiy Mosgprnd p } y ag700 la 3M EFnvoirrmonm1e5nt4a.l8L0aboratory 3M Environmental Laboratory (et Sty DcDxevoiagtieoneNo. 0_03 TR) Page 221 GAR: Report E01-1245 PFOS: A Reproduction Study with the Nother Bobwhite Record of Deviation 1. Identification Study/ ProjectNo. 01-1245 - D(eCvhieactkioonnTey)pe GPrSoOtPocol 00 OMtehtehr:od O Equipment Procedure Document Number: ETS-6231.1 Date(s) of occurrence: Throughout the study "1 Il. Description: Required Procedure/process: ~ ~ Sdeecatciotniv11a.t3,eandSd,enrteegnrceess5iosntarteecsa:lcuAlnatyed.l.e.vel outside 75% - 125%~of nominal value must be. ActualProcedure/process: o_o `aMpaenxyooffthtehecuhrivgeh.caWlhibernatiaoncaploiibnrtatsioinn tphoiisnsttluideysaarbeovkeepttheinatpheex,cuitrvisedceosnpsiitdeebreedinigndaebteorvmeintahnet (flagged with an T) and no concentration is calculated for the standard. If no concentration is calculated, no percent difference `within the stated criteria. is calculated for evaluating whether or not ~ that standard is `Removal of these points in most cases causes the next calibration point to then be above the apex of the curve. This continued deletion of points could continue until there are no points rraenmgaeinoifntghoencutrhevecuwrivteh. nToheevraelfuoartei,ocnaolifbpreartcioenntpodienvtisataiboonvcerittheeriaapex were included in the Ill. Actions Taken: -- "Tis deviation was wri_ti(esnu,cahanostaemteonfidlmeewnatsiwsrsiutetde,n,SaOnPdredvaitasipooni,nettsc.w)hi_ch__we_r.e_a_b_ov.e_t.he__ next highest calibration point were reanalyzed o |RecordedBy LisaA. Stevenson Olin A, Shusion snd Authorized By Mane (Study D'irector be /Project Lead) Spun Represtatobiee" AEfa dod Clik. "Dae 08/16/02 Date 8/zofozs 3MFEonvrirmoEnmTeSnt4a8lL0aboratory 3M Environmental Laboratory sidSdyDioDaePviraetiLoenndNo.5_cd_o1_d_y o_p_ro_ Page 222 rsa Report E01-1245 PFOS: A Reproduction Study with the Nothern Bobwhite Record of Deviation 1. Identification Swdy/PNro.oEjO1e-1c24t5 T TTT |DeviationType ~~ VSOP 0 Method O EquipmentProcedure | (Check one) O Protocol 0 Other: Document Number: ETS-8-231.1 |Date(s) ofoccurrence: Entire Stud, IL. Description: SRceiqouni1r2.1e, CdaPerlocoendsu:re/fporrobecsecss:elaevts ithanothonse issdheyvill document i ecwcts. A`(Scttunu5daadlPE2OrIo-nc103eo0d52u0m3r57eL7/1op,pfrproe)ocpedshses:tr.dTsis_p__i_"e_1d_sprpscsornecsiFvBpoi.nt" FEwis pos pSome.waS:ki i. of0200-- 1 57 See mLoutof4OmL. ieeddnsedr ewpindedoopngeSrPE.schcralrcinsea3d0,m.J4oDfMoelIOHe.SmTHoEfRsEFrORSEh.ed. Aber 1o0 fitd. sen (502008 POS mlMOH) * dl)(10mL/000L) * 1000/=2100g0PR)OS sto SPEcolumn {2TSh0e0isgeePRhreOsSs2ean.od0nLeMcEaoOrHsdbe=di110n9mPpeFpsOeSshneks.cnoMdenrOeepHo.niso 1h0e0cghPoEmOiSonglPMEGQyHe)r SainmgiPrRyOSh!e1fo0llmow.inlg tstdatsmeennt is(r4e0,.0. iedser 2Or.iii se) = 00mgPROS ml.Sera 1heisslenansgPtROSshowSsetrhfeofmthe 4chDro(omrat2o0grmaphMErOsH 0.ha05.nSzn)FiOdS".cMmOcH". he unis whencallin he nl SCEharDmoTmpac1tC2og8ara3lp0h4ai8c1oe:4l54s-1028972,n1/0pnomMfemOsH:, apdoitn,d0v2a.l0ue1092 10 np.Sec. Sic the 0. nil srevolumeof e(2r9.i20i5tPRdOSwith301m40.0m.MoerwOaaHt)We,on(2dd.i0lmamiboMn1eO5GRD/L0AorS05h1e.e1JSsncelrees=S1PE7p0rnegcPeRdeOsS.tecmiu.lSatsion 5 allows MSSaemiOamHleeCsa"pEkDleInd|i3io4nt536m1k55,itGeCd1s00rppob-O105TmhQoCisiatmsplsee)in.prepared by dog 2a. of 300152 (5.030 PROS: mL. (50209) Ga) * (LOA) (1000nghg (005 mi.Ses)= 20083g FOS i.Sra Usingthcarclulion, the rly cstof 6.17ng/mL.clues 0246Sm.Sr.Sinethe pike evel 200.8ngPEOS'/mi.Sera, the recovery is(246.8200.8)(100)=122.9%. -- -- (suchas amendllml.eAncttiisosnuesd,TaSkOePnr:evisionetc.) TE RLeicsoarAd.eSdBteyvenson (YAA. Sh3 uenson- - 7 Dae 108716002 ogifo, at Aes ec Authorized By (Study Director /Project Lead) AO ee Date Shay 0 Lbrae lony Moy sovement : h < AY TA.DeviationNo. _____ 3MEnvironmental Laboratory Page 223 GOLLOG Report E01-1245 PFOS: A Reproduction Study with the Nothern Bobwhite Record of Deviation Study / Project No. E0I-1245 1. Identification Deviation Type (Check one) osop V Method 0 Equipment Procedure 0 Protocol O Other: Document Number: ETS-8-231.1 Date(s)ofoccurrence: Throughout the study Ti. Description: Required Procedure/process: `Section 13.3 of the method states: Two thirds of all quality control samples and 1/2ofcach quality control sample at each level are expected to show an accuracy of 75-125%. AAc1t2ua5lppbProcqeuadiulrel/ivperroqcueaslsi:ty control sample recoveries were >125% Io @us oles Te 03) es An3loy Ti, Actions Taken: ~~ (such as amendment issued, SOP revision, etc.) ~ This deviation was written, liver data reported at the low range of the curve may be biased high, and quality control data not within criteria are flagged in the raw data. RLeiscaorA.deSdteBvyensonOf A Sector Authorized By packs AE? ~ (Study Director 3 /Project Lead) 90M LaborabyMascgemants pe tolls fagAy D0a8t2e6/02 #2L0 a fo Date 3M EFnvoirrmonEmTeSnt-a4l8L0aboratory 3M Environmental Laboratory gus (aed SyDicDevigactiLoesndNo. s_n_oBuyLrt) Page 224 GOL Report EO1-1245 PFOS: A Reproduction Studywiththe Nothern Bobwhite Record of Deviation Identification Study Project NoE01. 1245 D(eCvhieactkioonnTe)ype PSrOotPocol (1 Mtetehrod 1 Equipment Procedure Docomen Number ETS.8.231.1 Da2t/e1(2s/)02of1o3cc0u2rr.en2c1e4:/10/231/20128,020/21./0421, 2171220,24,1267100022, 211/02, i. Description: wRiellqubierdeodcuPmreoncteeddurien/tphreorcaewsdsa:ta.ection 12.1, Calculations: othe calculations a used than thos itd, they "ATcotueavilnPiornocuepdrusrdee/spdroeceissn:#1 clinghe ve lets: S0t0anpdamrd.E)01i1i3o6|2m-L3o6f0v1Prr0ep,hsoemtagdeesc.riptTihoins"hCoumrovgeePnaotne cpongosif$ir03s5,3tscrprppue4d5bOy spmikit.ng2r0SoocfCI0um01t6of SSO5053S3AmoLf. hivoem)o"gspekedu.sdA1RpTrsshcohmoxgternactied ssapnikderd,n1d5p0roefprheed5u0s3in53SP(E0, 0t0h6e3f5a8.l clo3t83 3 1000 ooffMwemtOH+. THEREFORE. 2001) (100 ug PROS MeOH) * 1.0 mL/*1(1000050350)= 120 1PR)OS oto SPEcoma |2S0i0n0e5 PROsSt'an2 dardm1 Lr0 McoerOdHd =311000pn,g PheOoSr.c MnetOsH.se1P 001gOSofM, OH) Ths re he unt bing mesureapore n he chomaagrapve sym. Simih,e ollowing socment is re. (2008 PROS 110m booger)* (1.0m homoge 0 1003S give) = 199ng POS g vr S"EaOm1-o3C4o2l-c3a0,p5pm4a5.3M3e3r" i CRohrroogresioagnadpwiiehe8s0tmsv10e7,6 nsng10pmoireedvalfoerst2h09ernel.SSPiEnperothce s3,3th8e Hnialosampo le wewiogfhnet i 10338 0.0m wate+ 10738 give)= 01030f5 ve in1.0m ofomogense removed fo extraction 010760 PROS! ml. McOM)* (2.0mLMeOH 0.103032 iver) =209ngFOS vr `SSMaiemmOppHll)eeC"sEplDiIkead3nn42t:36Imi01v7 e1QC0ho5mo5ge1na"te ThQCisamsple rbp yslia ng4url.ofd 0201-52 (5 0205PFOS /mi. (6*)(5020/1) * (LO *(10L 0ngi) g 0 1063Sg ive) = 200of FOS iver |UPsiOngSthgeicvaert.erthceolelcaotvioeny.it(h27a02n00y)a* 1s00u1o1f3l1503%9mg. call1s270ng ive. Since th spike eel is 200mg 1 (such as ameInL.dmAecntiisosnuesd.TaSkOePn:revision ec) | Issuingofhis SOP deviation { Recorded By Dae |1 Lissa sivenson Yo A Sleynion 12002 oily | Authorized Bboy (StSunudy DiGrelctoor /Project LeaJd)|ol Dae Won SCHrEnyviinmReonigliLaeborratoOryho +H 7 7 57a DeviationNo. __3___ Form ET5.4.00 EndyStdDirectora ProsLead coo dy orrot 3M Environmental Laboratory Page 225 Report EOI-1245 Repo E1245 PFOS: A Reproduction Study with the Nothern Bobwhite PFOS: ARopSrtudoy wdihnueNocx tBoitne eree eee ene eneespesmreeeeeeeeeemmet ATTACHMENT F: EXAMPLE CALCULATIONS --_-- FORMULA USED FOR SERAANALYSES IN STUDY EO1-1245 AR (ng/mL MeOH) x DF x Diluton(diued sera) = ng/mL. Sera Colculaton Used fo EO1-1245:20481, 454-108-370, 10 ppm Female, adult (inital volume = 0.3 mL) 220.24 ngimL MeOH x 1x40 = 9170 ng/mL Sera -_-- FORMULA USEDFOR LIVER ANALYSES IN STUDY EO1-1245 AR (ng/mL MeOH) x DFx`amounoFtifnaIlveVroilnudmieute(mdi sample (g) = ng/g Liver (Calculation Used for EO1-1242:30672,10 ppm, a.369M Liver (iis weight =1.0010) 39321 ng/mL MeOH x 10 x 20.1000m0L90MgeOLiHver = 78500 nglg Liver -_-- DAFR == DAinlaultyitoincaflacrteosrult from MassLynx summary Diltion(iuted sera) = Final volume(mL)/amounotf saa i died sample(mt) Amount of erain cuted samplo(rL) =amouofnsetra sedi Mota amount ofsera + watt) hAommoougnetniofzivng(imonid=lgr)ued sampl(c) = amounotfver weighed(c) amount of Iver weighed) +water added for Examples: AmDoituonnt(o@fisueodasin euteed)sam0gl1e(vnoLu)m=ea(m2o.0unmtLo)fasmerouunseofd(s0or.a3imnL)iAtcetadlaammapunlto(f0s.0a5rmaL+)a=t4e0r(6mL) = 0.06mL. aAdmdoeudntorofhveorminodkgteed(ns0aa0mnmLeg))== a1m00o1n0.9of/I1v0er60w1e0g5he=d (11..000010000a4mount of verweighed (1.0010) + water _-- Sootr:a fFooarscooans.amFpole ewshslessaiasn, 0a5chsinrdohcads,oaalhoatlwivstUoprGwaaesdad1d0ihn rercfoonrsemGanh10 of tod F"oarngxe+a00n2M3UsAaOmMiLw~h003 mLedwih0.02.ser.vite, 0.98mi.of airvasadded the mounof era i tnd Theditosernait)(=2a0 UuD02tmL.d50 WM Envonmontl Laboratory 3M Environmental Laboratory. 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