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TOXICITY TO AQUATIC PLANTS TEST SUBSTANCE Identity:A mixturecontaininpgerfluorooctanesulfonawthei,ch may alsobe referretdoas PFOS, FC-95,or as a component of FC-603EF. (1Octanesulfoniaccid)(CAS # 2795-39-3). Remarks: The 3M productiolnotnumber isnotnoted.The testsample is FC-603EF. Currentinformatioinndicateistisa mixtureof 1.30% PFOS, 10% diethyleneglycolbutylether,54.15% water,22.5% ethyleneglycol2,.6% propanesultonefoamer,5% Alkylethesrulfate0,.1% N-(3Chloroallyl)hexaminicuhmloride0,.9% Xanthan gum, 0.9% starch,1.0% triethanolaminaen,d 0.05% tolytlriazole. The followingsummary appliesa mixture withincompletely characterizedconcentrationsof impurities.Data may not accurately reflecttoxicityof the fluorochemicalcomponent of the testsample. METHOD: Method: OECD Guideline201 Type: Acute static GLP: No Year completed: 1996 Species: Selenastrumcapricomutum Source: In-housecultureO.riginalflryomATCC, RockvilleM,D Element basis:Algalcellcounts(cells/mLa)nd averagespecifigcrowth rates. Exposure period: 96 hours StatisticaMlethods: IC50 valuesdetermined by the Trimmed SpearmanKarber Method and Toxstatsoftwarepackage,Version3.5. Analyticalmonitoring: pH and temperature. Test Conditions: AlgalNutrientMedium: Per OECD method. Thisnutrienmtedium provided allmineralnutrientesssentiafloralgalgrowth and alsoserved as thediluent forallalgaloperations.The pH ofthissynthetiaclgalmedium was 9.06. Stock and testsolutionpreparation:A 1000 mg/L primarystocksolution was preparedby dilutin1g50 mg oftestsubstancein150 mL ofstandard algalnutrienmtedium. Aliquotswere thenadded directltyodilutiownater in testvesselsto createtestsolutions. Exposure vessels: 125 mL borosilicate-glEarslsenmeyerflaskscontaining 50 mL oftestsolutionand capped withfoam stoppers. AgitationS:hakencontinuousalty100 rpm. Number ofreplicate3s:forexposureconcentratio6nfso,rblankcontrol. Initicaelllloading:1.0 X 104 cells/mL(1.2x 104 actual) Number of concentrations:fiveplusa blankcontrol Lighting:Continuous400 ft-candieussingcool-whitfeluorescenltamps. Water Chemistry: pH range (0-96hours): 7.48- 9.06 Test temperaturerange (0-96hours): 24.2- 25.8'C RESULTS Nominal concentrations: Blank control,31.25,62.5,125,250, and 500 mg/L Algal Growth Response EC50 values Exposure (Contact) Hours 24 48 72 96 Cell-Count mg/L (95% C.I.) 139 (117- 165) 153 (124- 189) 188 (173-204) 141 (123- 162) Average Specific Growth Rate mg/L (95% C.I.) 171 (146- 199) 290 (254-330) 275 (252 -300) 294 (265 -326) Element values are based on nominal concentrations. Reversibilityof Growth InhibitionA:liquotsofthe 500 mg/L testsolution were dilutedwithalgalmedium and culturedfor7 days. Based on growth observed inthe recoveryphase,the effecton algalgrowth was found to be algistatic. Remarks: Testingwas conducted on the mixtureas described inthe Test Substance Remarks field.The valuesreportedapplytothatmixtureand not the fluorochemicalproportionalone. CONCLUSIONS The testsubstance exhibitsa 96-hourEC50 cellcount value of 141 (123 - 162 mg/L) and a 96-hourErCrogrowth ratevalueof294 (265 - 326) mg/L. The 96-hourNo Observed EffectConcentration(NOEC) is31.25 mg/L forcell count and 125 mg/L forgrowth rate.Thistestsubstance was determined to be algistatic. Submifter: 3M Company, EnvironmentalLaboratory,P.O. Box 33331, St. Paul,Minnesota,55133 DATA QUALITY ReliabilityK:limischranking= 2. Thisstudymeets thecriterifaorquality testing.However, sample puritwyas notpropedycharacterizeadnd thestudy lacksanalyticaclonfirmatioonftheamount offluorochemicaplroportioninthe solution. REFERENCES Thisstudywas conductedby AScl CorporationA,Scl-DuluthEnvironmental TestingDivisionD,uluth,MN attherequestofthe3M Company, Lab Request number P2581, 1996. OTHER Lastchanged: 6/26/00. AS,:[C,,rl%,rAts.,ii,AS,:[-Duluil@ I'n%iriiiiiiTi%c:i.i@ttiiDtnig%isititt AS,,:lStud.vIDJR 5010-Obg STUDY TITLE GROWTH INHIBITION OF FC-603EF. LIGHT ATC 3%, AR-AFFF TO GREEN ALGA (Seletiastrciaipir?it-oriiiiiiiiii) DATA STANDARI) 01--CD GUIDELINE 201 STUDY CO,%IPI.EI'Et) Noveiiiber22. 19k)O TESTING FACILITN' AScl Corporation,ASc-iDuluth EnvironmentalTestinC,gDivision 4444 Airpark Boulevard Duluth,NIN 55811 Tel.No. (218)722-4040 F ax N o. (1.18) 7'@)-.')5-9)2 STUDY IDENTIFICATION NUMBERS AScl Study ID# 5010-068 3M Company Lab Request# P2581 S;)onsor. 3M Company SponsoSrtudyIDIP".581 AS,.l Efl,ir,wnitici,.TtLa:t,EitilD:t%i.i,,n AS,:l St@,dl@ll)g 5010 068 1.0 GENERAL INFORNIATION Study Title Data Standard Sponsor Sponsor's Representative Sponsor's Lab Request# TestingFacility PrincipalInvestigator ProjectDirector TestingFacilityDirector DefinitiveTest Dates Data Validationand Report Review Growth Inhibitioonf FC-603EF. l.iahtWater (TM) ATC 3%, AR-AI:Fl:to GreenL AIL--A (Selenastriicta?piriet)riiiiiiiiii). OECD Guideline201. 3NI Environmental Techiltolt),a,i,l@dSer,,,ices. Building2-3E-09, 935 BusliA%,cntie.St. I'aLli. MN 55133, Tel No. (612)778-7452. Susan Beach. 3NI En%-ironniciiial and Services,Buildine"-3E-09. 93i liLISI) Avenue. St. Paul. NIN i@,133: Tel No. (612)778-7452. P258 1. AScl Corporition,@ASci-I)LIl1L-I1t)iI'lrt)[Illlelltil TestingDivision.4444 Airl).trBkotile%iirki. Duluth,MN 558i 1: Tel. No, ("18) 722-4040. Todd NlcGuire Joe Amato Donald Mount Novei-nber4-8, 1996. Alan Niozol Report Si,,iiattire: Retentionof Raw Data and Final Report Location of Raw Data and FinalReport Two years from thedate study iscompleted. ASci Corporatioti/ASci-DtiltEintvhironmental Testinc,Division,4444 Airpirk Boulevard. Dulutoh,MN 55811; Tel. No. (218)722-4040. Sponsor; 3M Company SponsoSrtudyID# r-581 2 @%S. I Si...l@ ll)z 50!0 L)68 2.0OBJECTIN'E(S) To determinethe4-d medianeffecctoncentrati(oEnC,,,t)h.e4-dno obser,ableeffect concentrati(oNnOEC), andalgicidal/asltgaitiecffecotf thetestsubstiiii1c:e(.'-w3E[:. Water(TNI)ATC 3%, AR-AFFF forgreenalga(Si,lt,ii(its-itritpirni?(-ortittiintdtelrsit?i)l)ltiecst conditions. This study was conducted accordingto OECD Guideline201 (OE('D 199-1). I'cstSubstance Test Oroeal nisms Algal Medium 3.0 TES'I'NIE*1'110[)S Properliev: Aiiiber-coloreldiqtii(sip.ecificEriti-t,%ct. 1.0t) 1). stable,and conipletelsyoltit)iliei Te.vctoticentrafioit0s:(a]-2allllCdlLiclolyliirol)3. 500 mg/L. Soiirce:ATCC 22662 hiocultim preptiriiiiott: Al-al stock was Subcultured 4 days before test initiatioii. iii01:Ct) ille(iililliI*tlr U.ve itt test: Stock culture %%-as dillited and spiked lilt()test SOILitioll" to obtain an initial cell density of 1.0 X 10, cells per nil ( 1.2 N IC)4 actual concentration). Preparatioti:Deionized water %%-asspikedwitlinutrientstocks(includ[112 NA,EDTA) to withinOECD recommended concentrationsT.he most recent chemical analysisfor deionized.-.ateirsin Raw Data Packat-,e. CliaracteristicsA:t testtime, the mediuni had a pH of 9.06. Sterili/,%e-dia autoclavepriorto u.,ein the test. Sponsor. 3ArtCompany Sponsor Study ID# F"-581 3 AS,*lC,irrk,r:ili,5ti,AS@l-Duluili En%,ir@,iiTicnsetntnaDgli%isii)n AS,:lSitid-1@[)#5010-069 Exposure Chambers Incubation Observations Endpoint Calculations 7:vpe:Sterilize1d25-ml borosilicate-glaEsrsienmeyer flask-s,eated%vith foam stoppers. U.veiiitest:50 nilof algalniedium or testsolution/chaiiiberS.ix control replicateasnd threereplicatepser testsubstanceconcentration'A-ere employed. Dufinc,the test,keptstoppered,exceptwhen experimental observationswere made. Solutionswere not aerateddurinc,t,he CXfX)SLires. Ditration:4 days. Dailypht)t(perit)iClo:ntinuousat 400 ft-candieussingcot-)]l-i,i.i@e fluorescenltamps. 'restchambers heldon shakertabLleset itI(X)ri)iii. Tei?ipertitt2i5r.e0: 2@C. Biological:Daily cellcotitittsisin-d,irectmicroscopicCilLiii,,cri%tit-i,Liiii Improved Neubauer heiiioc\.tt)iiieter. @V(itecrliettiivi(n1.):At testinitiatiaonnd terniinall(Ii)iHi.-CO[ILILLCiItI\\, and temperature,(2)daily-temperature.All determination\%,kerieii:tde accordingtoAPHA itiethod(s1995). 1-,2-,3-,and 4-d EC,,,'sa.nd 4-d NOEC based on botilCellCOLlilainsd averagespecifiocrowt[irates(,u).Algicidal/alcistdaettiecriiiiiii\i%t,iatsiii made posttoxicittyesttermination. Sponsor. 3161Company Sponsor Study IDO P-'591 4 ASCI Dultitit Fnvir-inrnciTlct%jtlingDi%isiiin AS,:lSi,.dyIDO 5OlO-Ob8 4.0RESULTS Test SubstanceToxicity(Pertinendtataare inTables 1,2, and 3) Test Conditions CellCt)itt1i-td EC,,: 139 iici@@oil@(117-16-5). ti1-dEq,: 171 ingl,'(L146-199)- CellCot4ni2-d EC,,,:153 ii2-d EC,: 290 nig,'[(-2i4-330). CellCo"nt 3-d EC,,,:188 ingi'L(173-'-'10.-1 IA3-d Eq,,):275 mg,L (252-3(X)). CellCt)iin4i-d ECs,,:141 Iloi,I,-@(12 1t,,I ,u 4-d EC5,,: 294 iii-l@t- ('-'65-326). Ci,il Cotipit 4-d %70EC: ', I . I it 4-d NOEC: 12i iilc@o I.. Effectof testsubstaiic%e%;idseteriiiiiiteodtic algistatibcased on reco%-erovf cellsexlx)sedtt) 500 mg/L concentration. Tet?ipt-ratt(i'rCt),: L@@hiititeitsiry.ft-c4-(dXl)v.: pil: 7.48-9.06. Specificcondiictit-itv 137-15'-'. Spon;or: 3161,Company SponsorStudyID# PI-581 5 AS,:] En@ir,,iiii@viiiii AS,:lStuj@ 1[)#5010 OtS ';.T0EST QA/QC QA/QC Cri,,eria Algalcountsincontrolmust increaseby at leasta factoorf 16 duringthefirs3t-d of the exposure. Testdurationmust be 4-d. Data During thefirs3t-d of thetestcontrolal,,al countsincreasedby a factorof approximately 39. The testdurationwas 4-d. 6.0 REFERENCES AmericanPublicHetithAssociatio(nAPHA). 1995.Standard%lettitxfits)irtiIc:xaiiiiiiioili'(iii Water and Wastewater.APHA, Washington,D.C. OrganizatiofnorEconomic Cooperatiotaind De%-elopinet(i0t1-Cl)).IL)930.1--Cl)(iuilielitiLi)ers Testingof Chemicals.OECD PublicatioInnformatioCnenter.Washinc-tonD..('. Hamilton,M.A., R.C. Russo,and R.V. Thurston.1977. Trimmed Spearniall-Karb%eIretllod forEstimatingMedian LethalConcentrationisnToxicityBioassivs.Erl%,irkS)cri.Tc,,:Iltlol. 11(7)7:14-719;Correctio1n2 (4):417 (1978). West, Inc.1996. ToxstatSoftwarePackageVersion3.5. Spons.or3:NICompany Sp4)nsorStudy ID# P-1581 6 AS,:lC,)rlx)ratlAo&lIl Dtilutii Te@iiiii: AS,:lSt-d@ IDP 5010 0(,S TABLE 1. EC,'S, 95% ConridenceIntervalsa,nd 4-d NOECIS rrom AlgalGrowth Iiihibitioll Testwith FC-603EF Light Water(TM) ATC 3%, AR-AFFF Endpoint Cell Count Time 1-d EC@ 2-d EC;., 3-d EC,i 4-d EC,, 4-d NOEC Data- (mg/L) 139 153 188 141 31 95 % Ct)nl-iden,:c (nijl) ll'-I(,5 1 1'14- 1 EndFx)int Averai!e Specific Gro%%,IhRate (;A) Time I-d EC@, 2-d EC,, 3-d EC,, 4-d EC,, 4-d NOEC Data- 171 290 275 294 11-5 95'i @ll I i -3 NtitAppli --thl- Values Based on Nominal Test Concentrations Spunm)r: 3.kfCompany Sponsor Study [D# P-1591 7 AS,:[ -\S,iD,,[,,Ili AS,:lStiidkli),5/010068 TABLE2. Cellspertnland PercentagDeifferencferom\leanControlValueforSelenasti7itit capricorntilEuxmposedtoFC-603EFLightWiter(T.\I)A,TC 3@'tA,R-AFFF TestConc. (mg/L) Contrt)l 31.25 62.5 125 250 Soo Replicate A B C. D E F \Ie@in A 13 c .\Ie@in A B c %lean A B c .\lean A B c Mean A B c Ni can Id 8.5 (-1 8.0(-) - I-) 5.5 6.0 9.i 7.5u -T Cell.-i.('10r4)nl 2d 24 (-) 25(-) 28 37 27 1-7. -3 -1 49 t-) 4S 39 5.i 43 4(@ 4 1 8.5 (13)" 7.0 (-7) 8.0 (7 @ 7.83 (4) 17 i-19@ 1()1-ill -'2 (--,)1 ),)-.'(-.II) 5'4(1f 41 4(1.11 10 (33) 6.5 (-13@ 5.5 (-2,) 7.33 f-I-) 11)(-311 17 ( 401 22 (-2-1) 19,(@1-31) -45 -4) 4,;1-4) 44 -.i) 4.4.3(-4) 3.i 1-53) 1.0(-87) 4.0(-47) 2.83 (-62) 20 (-30) IS (-37) 15 1-48) 17.2 (-38) 43 (-S) 39 (-1(1) 3S (-IS) 39.S (-14) 3.0 (-60@ 2.5 (-67) 2.5 (-67) 2.67(-64) S.i (-69) 5.5 (-SO) 10(-64) 8.00(-71) 12 (-75) lu (-78) 9.5 (-So) 10.3 (-79@ 2.0(-73) 0(-Ioo) 3.0(-60) 1.67 (-78) 2.3(-91) 1.5(-95) 3.0(-S9) --).3(3-92) 2.0(-96) 1.5(-97) 1.3 t-97) 1.67 (-96) i (-I I I") '4 54 1 ;1 @l,) 4(,(-34) 4(,(-14) 40.' 1-3-') 14 1-St)) -1((1-72) 14 (-80) 1i.7 (-77) 2.(@4-97) 0.,)(-99) 4.0 t-94j 17 t-97) Control ReplicateC Values Omitted iL-a;n Outlier b ParentheticValues = % Difference from C4)ntrolNlean Value 0-d %lean Algal Count 1.2 X 10, ..Sponsor; 3,%lCompanv Sponsor Study ID& PI-@ST 8 ASci C,irpiriii,,AnSci-Duluth L-'ii%irtinritenTtci.l%tingDj%isj4iii AS,:lSluJ% IDZ 5010-0tS TABLE 3. Average SpeciricGroulh Rates (,ua)nd PercentageI)irrerencreroni%leiiiControoll Value forSeknastrum capricornutumExposed to FC-603EF l.ighl%%ater(l'.NAlR)AFFF Test Conc. (mg/L) Conin)l 31.25 62.5 125 250 500 Replicate A B C, D I.F ,%Ican A B c Nicaii A B c .%lean A B c Nlcafi A B c Mean A B c Mean 1.958 1.897 1 522 I,bw 2.069 1 833 1.959 (711.764 ( 4) 1 897 (4i I 8"t, 1-1 ]'-0(lb) 1 689 1-81 1.522 (-17) 1.810(-1) 1.070 1-42) 0.182 1-100) 1.204 (-34) O@859 (-53) O@916 (-50) 0.734 (-60) 0.734(-60) 0.799(-56) 0.511 (-72) 0(-Ioo) 0.916(-50) 0.329 (-82) .p = In Cells per mi t-- In Cell%/mi I t.- L, Control ReplicateC Values Oinittedas in Outlier ParcnUm:LicValues = % Difference from CtinirL)%llean Value 0-d Mean Algal Count = 1.2 X 104 Sponsor. 3161 Company Sponsor Study ID# F"-581 Avem@c Specific GnvAih Rite 4 r 2d 3J 1 @487 1.509 - I-) 1W f 1 707 1 547 1 '.S70 1 325 lb, 1 3tg 13) 1 454 1 385 i !Zt 1 233 189 I zis 1 381 12) 1,311 17) 1 443 81 01 1 @381 1 1 394 t-Ili l@340 ( 15@ 1 246 1-21) 1 330 1-15) 0,979 1-38; 0761 ( 521 1.060(-32) 0.949(-40) is') 1 iw 1 is, i@ 1 !C)7 1-4! 0 75 -,S. 0 707 '4'-) 0 690 4-431 0.718 (411 0.367 (-77) 0.112 t-931 0.458 1-71) 0.332 1-79) O@ 170 ( 9846,)) 0.074 1 0,074 10 1]01 4d 0 )7S 0 1)8.1 0 os-, 04@, 101! 0 ),)S I 0 U,017 S. o').-, 0 _, : E, 0,)41 71 0927 i 9 1 01)01) 10@ 0,)0-) 10, 0915 91 ot,], 0 o,)," I 0605 :-40)) 0.642 -3o 00,128 ( 871 030119 1-100) 1-70) 48 4-851