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ACTIVATED SLUDGE RESPIRATION INHIBITION TEST TEST SUBSTANCE Identity:A mixturecontainingperfluorooctanesulfonatweh,ich may alsobe referredto as PFOS, FC-95, or as a component of FC-203CE. (1Octanesulfonicacid)(CAS # 2795-39-3). Remarks: The 3M productionlotnumber was 519. The testsample isFC203CE (2/21/89).Currentinformationindicatesitisa mixtureof 1.84% PFOS, 30% diethyleneglycolbutylether,60.23% water,2.56% Sultone foamer, 3.1% sodium octylsulfate2,.1% polyoxyethylenemonooctylphenyl ether,0.12% sodium laurylsulfatea,nd 0.05% tolyltriazoAlnet.ifoamerSWS 214 was added ata nominal concentrationof 200 mg/L. The followingsummary applies to a mixture with incompletely characterized concentradons of impurities.Data may not accurately retlectthe toxicityof the tluorochemical component of the testsample. METHOD Method: OECD 209 GLP: No Year Completed: 1990 Analyticalmonitoring: Dissolvedoxygen concentrationsp,H and temperature. Statisticamlethods: Resultswere determined by calculatioonfthe % inhibitioinn respiratiornate. Test organism source: Activatedsludge mixed liquorcollectedfrom the Metro Waste Treatment Plant,St.Paul,MN Test condition: Dilutionwater: MilliporMeilli-(Ywmater Synthetic sewage: per OECD method Reference and testsolution preparation: A stocksolutionofthe referencesubstance,3,5-dichlorophenowla,s preparedby dissolving 500 rng in1OmL 1N NAOH, dilutedto 30mL withMilliporeMilli-TQM water,then broughttothe pointofincipienptrecipitatiwointh IN H2SO4 and dilutedto 1L withMilli-dTwmater. The pH ofthe reference substance was measured tobe 7.4. Test substance was added directltyothe testvessels.Antfoamer SWS 214 was added tothe test vesselsat200 mg/L from a stocksolutionpreparedby dilutin2g000 mg ofSWS 214 in500 mL Milli-lYwmater. The testsubstance and antifoamerappeared miscibleinwater. Test vessels: Not given Number of concentrations: threeplus 1 referencesubstance and 2 blankcontrols Temperature: 20 - 22 OC Totalsuspended solidsand pH on day oftesting5:.033g/Land initiaplH 6.6 and finalpH 6.9, InifiTaSlS of 1000 mg/L concentrafion was 2.027 g/L,pH 6.6 whilefinalTSS of 1000 rng/Lconcentrationwas 2.213 g/L,pH 6.8. Element Basis: Respirationinhibitioans determined by oxygen consumption. Remarks: Values correctedto 20 OC forcalculations. RESULTS Nominal concentrations: 2 Blank controlsr,eferencesubstance,320, 560 and 1000 mg/L Element values: 30-minute EC5o = >1000 rng/L 3-hour EC5o = > 1000 nig/L Element values based on nominal concentrations. Remarks: There was a slightreductioninthe amount offoam formed inthe 1000 mg/L exposure concentrationwhen compared to FC-203CE without antifoamer. Testingwas conducted on a mixtureas describedinthe Test Substance Remarks field.The values reportedapplyto thatmixtureand notthe fluorochemicalproportionalone. CONCLUSIONS The testsubstance 3-hour EC50 foractivatedsludge respiratioinnhibitiownas determined tobe >1000 mg/L. Submitter: 3M Company, EnvironmentalLaboratory,P.O. Box 33331, St. Paul,Minnesota,55133 DATA QUALITY Reliability:Kliniiscrhanking2. Testingmeets allcriterifaorqualitytesting. However, sample puritywas notpropedy characterizedand the study lacks analyticaclonfirmationofthe amount offluorochemicalproportioninthe solution. REFERENCES This studywas conducted by the 3M Company, EnvironmentalLaboratory,St Paul,MN, Lab Request number H1480-3,1990. OTHER Lastchanged: 6/26/00 Type TestAz4i%iAl-&ti ENVIRONMENTAL LABORATORY ACTIVATED SLUDGE MIXED LIQUOR LR. Dat Analyoz@;ZAi Source Date Obtained All q icto Awl- -c4--.-@hLd@4t& Date Used ign Mixed Liquor Suspended Solids (MLSS) g/L Adjustment g/L PH to.Lo ta,q Comments Test Substance Sample Description PC-?Q-x klikeek.CoKcektra*30K+Jel-jt@ Sample pHim@tVM_i 1_&.Lo-o Adjustment Comments L4L-4,Scif-1006=3qlL. +P--,+- @v@uaj 2..071-(QIL, n\AlaZLLU20owSIL) /I Reference Substance Sample Description.%%, VA;g%est COAMV*raeAtvt Sample pH ivi,4;A6l. Comments 10 "IL hdiustment Exposure Conditions Test Duration Test Temperature 1-0 -7 0c Comments Form 1002o.S.0. pWo Sample Description EnvironmentaLlab- Work Sheet OECD ACTIVATED SLUDGE RESPIRATION INHIBITION TEST #209 Lab Request No.6J1480----A, Page of S Date: Analyst: 9/IQ Tpgt SnliitinngPrpparatinn (2 zitSC@NtetUgAk4iQ)ai,,ne-icSy 1 2 rX@ L)M"ll":Q -,S@4 LA I.,414,'O@duA-Q U:u 8 2= A @ft^ Test Conc. mg acfgd mlmq w-eer ml syn. sewage mi micr6bial Total volume Ion 4- 7 Sg + lt)O() 4- Z5-9 + Z S + I W- -r 2-84 -ir-0 +- ILp 4:::kX4,Pv- ZV, cnvtwn kT- LV A-lk4lg Y-eAuc:i@cyL'%V%-&MC-XA-V&- Of--POAVvt-@6nd&ai dl.Lnl'tCaLft!idS.i@j'iL4 AQ S-0 0 Sol) ENVIRONMENTAL LABORATORY LR p@ye- 2io-f-b OECD ACTIVATED SLUDGE RESPIRATION INHIBITION TEST #209 RAW DATA Test Substance: /.qg W, Source of Activated Sludge Mixed Liquor:h@(4ne= Date Received: Date Used: Analyst 511!?1,9,0 5 IA/,-32 1 30 - MINUTE EXPOSURE - lest Substance Concentration(mg/L) 0.5 CONTROL I 1.0 11.5 DISSOLVED-OXYGEN CONCE TRATIONS ma/L TI@IE-MINUT)S .0 2'.0 2.5 13.0 3.-9t 4 14.5 5.0 LS @?. ,5,4/ TEM r,.5 rIc 1 2,, zo L4 ?-.n CONTROL II Z,A/ Z,O 3 - HOUR EXPOSURE un lest Substance Concentration(mg/L) 0.5 CONTROIL i!i,9 DISSOLVED OXYGEN CONCENTRATIONS E- INUTES 1.0 11.5 2.0 2.5 3.0 13.5 4.0 (a Z, 4,a AIS 4 / A.7 mg/L 4.5 R 5.0 -.5.5 ;?,,4,/2,0 TEMI C ZL ?@2 -j5.o -r-@100 AA-A 9 i ,.7 2---7 7J 100c) ,Os6l,t;l_5o,;1J0-Ib4.A 6@ 0 CONTROL 11 19 15, 5 -@T, zz pH of Activated Sludge Mi%ed Liquor: PH of highest concentrationtestcd: Test Temperature:W-ZZOC Initial(.O.(D Initial (,.p.Lp Final(p.9 FinalLoilb ACTIVATED test SubstanceAr-'MrlCi:j EWIRONME-NTAL LABORATORY SUJDGE RESPIRATION INIIBITION TEST #209 CALCULATION OF RESULTS )with,4n6 @rc)a,,Z -Wn5e,rz 30 - MINLITE-EXPOSLJPE- Test Substance Concentration(miz/L) mmmnl,I- Oxygen Uptake Rate R - C).AL4gi R Corrected(') to 21fC PS IIto LR @IIAIW-.;S Date -;rJ,lc?lql)Analyst a4e.36@@ ' I Inhibitionin Respiration Rate(2) CONML I1 5; 2,--JSO Percent Differencein Controls(3) W@o (9S% Confidence Limits) mg/L -EXMQTRR Test Substance Concentr;@tion(mg/L) Oxygen Uptake Rate R CORMOL I %2 ":sI O.s ()no o. IRI) I R Corrected(') to 21fC % Inhil)itioinn Respiration Rate(2) (N.l(@041 1I-tp CONML II C).F)IOC)ia Percent Difference in Controls (3) BCSO(gstConfidenLciemitsm)g/L 2j,.Z@@6 (1) Temp. CorrectiiDn= o@ (mg 02/L-min.) RC1 + RC 2 (2)1 Inhibition RS x 100 RC 1 + @2 llhere: 2 R a Oxygen uptake rate (mg 02/L-min.) T obs - Smnple tem% Oc &I- 1.047, x - T,o's'- 2eC C - Control (3) IRC RL;ix ifference= lob. RC1 +tRiC22 2 RC The0e two cownnittrol re1s5'1piio@r+fatioodn@nor@aete@s@ (sh@suhlodutl@dbeb@ecgwiint@h1in 151 ofteiaoc@h other@' Type Test At,4.@LvA@--A ENVIRONMENTAL LABORATORY ACTIVATED SLUDGE MIXED LIQUOR Date C) Analyst::;@Z-% Source 0 IA Date Obtained -,-IrZ5-V>8 Date Used -tlr Mixed Liquor Suspended Solids (MLSS) SS-ic@ g/L Adjustment g/L PH t,,-l t..Ck Comments Test Substance Sample DescriptioA-'SAAL@'ZIJ44W@fL@ %N!3htbi- Sample pH _A@AW jai 1#,-l Comments Adjustment Reference Substance Sample Descriptiork_ kijumstcw"vr4ye"(M Sample pH la@4,-A6,,g -FIIAA-1 4. 0 Comments e AIA.OIAIJ Ovd[L. Adjustment Exposure Conditions Test Duration 0 Test Temperature c Comments I FoirnIOM - 0 -PWO SampleDescription EnvironmentalLab - Work Sheet OECD ACRIVATED SUJDGE RESPIRATION INHIBITION TEST #209 Lab RequestNo.U14RO-15. Page I of -",) Date: Alz,.A Qo C==:@ i Analyst: Stock and Test Solutions Preparation IJL@Ay-r@xr-A eir (411190 STOCK SOLURION: n@itiomes Q=p==-t6evA Yr, L zt4 /,-:@CV)'nWI- S 2J ve-YIU m "CSIhPi-e1,) Ady,@kam 4MQ tdi. Test Conc. ml stock soln mIM2 water kg/L per SOO ml per 500 ml ml syn. sewage feed per SOO ml 12,5 + Isci + 14 ml microbial inoculun per 500 ml 2 OC) Total volume ml LR 14148on5 ENVIRONMENTAL LABORATORY OECD ACTIVATED SLUDGE RESPIRATION INHIBITION TEST #209 RAW DATA Test Substance: Arl+;Rk-t@nce(141'ili;nwi)l Source of ActivatedSludge Mixed Liquor:I&tbm \&IA Date Received: 612-2,LqO Date Used: Analys 30 - MINUTE EXPOSURE iest !iUbStance DISSOLVED OXYGEN CONCEttTRATIONS ma/L TIME MINUT S Concentration (mg/L) 0.5 1.0 1.5 12-.0 2.5 13 4.0 4.5 5.0 S. c CONTROL 1 4.1 4.2-,--A.17,.l 2-.U i.q 1&5 ().f@ @l:cm CONTROL 11 5; Lci- 2, 1-ci I.L@ O.C'4 3 - HOUR EXPOSURE %a lest Substance . DISSOLVED OXYGEN CONCENTRATIONS ma/L TIME-MINUTES TEM Concentration (mg/L) 0.5 1.0 1.5 12.0 2.5 13.0 3.5 14.0 4.5 5.0 l@.5.5 Oc CONTROL 1 3 11,04.9 4.4 Zllg z; 2-,2- 1,S I-8 CONTROL Il Ts.5 !4.ci pH of Activated Sludge Mfxed Liquor: pH of highest concentration tested: Test Temperature: 18@190C, Initial Lv.'l Initial Final to.9 Final to-O O.S EWIROW-NTAL LWRATORY Test Substanc ACRIVATED SUJDGE RESPIRATION IMilBITION TEST #209 CALCULATION OF RESULTS c=Q=I-jevi, 30 MINLME EXPOSLJPX. LR -141LI-P@n@ Date rq4 Analyvst pa,gc Adyiavl, Test Substance Concentration(MR/L) MNM()T. T - Ionn Oxygen Uptake Rate R 1.6-164 R Corrected(') to 2e C 1.1AriA Inhibitioinn ----RespiratioRnate(2) OONTROL II ql PercentDifferencein Controls(3) Ecso (9S% ConfidenceLimits)mg/L Test Substance Concentration(mg/L) CONTROL I - Oxygen Uptake Rate R -- I.Iq.5q 1.114-N-sin R Corrected(') to 2CPC I.2,cA7-j I -Z-10 1 % Inhil)itioinn -RespirationRate(2) ODNML II Percent Differencein Controls(3) ECSO (951 ConfidenceLimits)mg/L (1)'Temp.Correction---Itr.s (mg 02/L-min.) 1 + RC2@ (2)1 Inhibition 2 /- RS - x 100 1 + RC2) llhere: (!C- 2 0/ R a Oxygen uptakerate (mg 02/L-min.) T,obs . le tap.,*c & - 1 ;47S7P x - T,obs - 2eC C - Control IRC, RC (3) % Difference- x lob. RCI + RC2 ThOeu:tlwo coonn@trolrle@s%dii)rifartaoinonraotaetse ss!hhoulwdtdohbeciwni@rtceh)ialncIhA ofteiach othheserr