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ACTIVATED SLUDGE RESPIRATION INHIBITION TEST TEST SUBSTANCE Identity:A mixturecontainingperfluorooctanesulfonatweh,ich may also be referredtoas PFOS, FC-95, oras a component of FC-203CE. (1Octanesuffonicacid)(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% tolyltriazole. The followingsummary applies to a mixtum with incompletely charactedzectconcentrations of impurfties.Data may not accurately reflectthe toxicityof the fluorochemical 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_QTwMater Synthetic sewage: per OECD method Reference and testsolution preparation: A stocksolutionofthe referencesubstance,3,5-dichlorophenowla,s prepared by dissolving 500 mg in 1OmL 1N NAOH, dilutedto30mL with MilliporMeilli-QTM water,then brought to the pointof incipienptrecipitatiownith 1N H2SO4 and dilutedto 1L withMilli-Qrwmater. The pH ofthe referencesolution was measured to be 7.4. Test substance was added directltyotest vesselsand was apparentlymisciblewithwater. Test vessels: Not given Number of concentrations: threeplus1 referencesubstance and 2 blank controls Temperature: 20 - 22 OC Totalsuspended solids and pH on day of testing: 5.033 g/L and initiaplH 6.6and finalpH 6.9. InitiTaSlS of 1000 rng/Lconcentration was 2.020 g/L,pH 6.6 and finalTSS of 1000 mg/L concentrationwas 2.200 g/L,pH 6.8. ElementBasis:Respiratiionnhibitaisondeterminebdyoxygen consumption. Remarks: Values correctedto20 "C forcalculations. RESULTS Nominal concentrations: 2 Blank controlsr,eferencesubstance,320, 560 and 1000 mg/L Elementvalues: 30-minuteEC5o=>1000mg/L 3-hourEC5o = >1000 mg/L Element values based on nominal concentrations. Remarks: Testingwas conducted on a mixtureas describedinthe Test Substance Remarks field.The values reportedapplytothatmixtureand not the fluorochemicalproportionalone. CONCLUSIONS The testsubstance 3-hour EC5o foractivatedsludge respiratioinnhibitiownas determined to be >1000 mg/L. Submitter: 3M Company, EnvironmentalLaboratory,P.O. Box 33331, St. Paul,Minnesota,55133 DATA QUALITY ReliabilityK:limischranking2. Testingmeets allcriterifaorqualittyesting. However, sample puritywas not properlycharacterizedand the study lacks analyticaclonfirmationofthe amount offluorochemicalproportioninthe solution. REFERENCES This study was conducted by the 3M Company, EnvironmentalLaboratory,St Paul,MN, Lab Request number H 1480-1, 1990. OTHER Last changed: 6/26/00 Type Test igr+iv4i-r-I ENVIRONMENTAL IABORATORY ACTIVATED SLUDGE MIXED LIQUOR H16+M L Date,-x.@/i/cqiO Analys4 tmkz- Source tap;hm Date Obtained --;sIl(RIgo Date Used Ici Mixed Liquor Suspended Solids (MLSS) ,5t)AY g/L Adjustment g/L pH )n.Ci Comments Test Substance Sample Description vu 11 Sample, pH loo. t2J2J.19,R) Adjustment CommentsMLSS nC-Jr)nC)wv4L4e-@-,M+vi-,14 !LA&-ViALI 2,n2-qlL-j,-VLvlAltaurL).2-8/L, Reference Substance Sample DescriptiorLA.";-.-NlclllnynDbt--rk,@l wgv,e.s+ Sample pH ilk,'-4;tAoi,"l ibm/L Adjustment Comments Fxvosure Conditions Test Duration Test Temperature 0c Comments Fo#snSV620 O-D PWO Sample Description EnvironmentalLab - Work Sheet .i OECD ACRIVATED SLUDGE RESPIRATION INIIBITION TEST #209 Lab Request No. P 1%R4O- Page of .5 Date: 9 q -o Analyst: Tpqt Iqnlutinng R3=aratinn Cl@;Ln+--Slg u i misc lhlrw,i-fiwiater, ce appmt-Awv-.: eleay-We-lier.A-"mw Test Conc. mg added MIMQ water ml syn. sewage ml microbial kO 2-B4 -ir. Ito 7 pr@; k- z-84 fto i-non' -Sim Bi@Ounz6ml-t 0 294 (a ZR4 + i(a i- 7rx,@ +- 2L 0 + mo Total volume ml -ti@m non Nno nilar ea-13i:@CO- 3L W*i-@-Ihg w 14i:kn- V4,v1--iw-g-@amw@j @ Lame.i arnnukni- t-,+-Poam -Pnn-ned, U,--.@ MEL, LR ENVIRONMENTAL LABORATORY OECD ACTIVATED SLUDGE RESPIRATION INHIBITION TEST #209 RAW DATA Test Substance: /-)a/-Yc?L21,z@lf Source of Activated Sludge Mixed Liquor:e,,!@m kras6-- Date Received:- Date Used: Allql,9,6 Anal 30 - MINUTE EXPOSURE iest Substance Concentration (mg/L) 0.5 1.0 11.5 DISSOLVED OXYGEN CONCE N TRATIONS-ma/L TI.M MINUT s 2. 0 2.5 3. 0 1 3. !1,-.4.0 14.5 5.0 CONTROL I "SIZO & -n 15.-q-,5,A/ .:5.0 v ti V 4,10 /,'g 4,5 A. 7 -j;. 11.9 -pstor.) 100c) z: TEM 5-5 rIc @@C -M CONTROL 11 5,8- 5,3 54,9 lov@4 1 t 8 2,011,0 3 - HOUR EXPOSURE lest substance Concentration (mg/L) 0.5 DISSOLVED OXYGEN CONCENTRATIONS ma/L IME- INUTE 1.0 11.5 2.0 2.5 13.0 3.5 )4.0 4.5 15.0 CONTROL 1 -,5, (a 5. L 4. q AA,.5 4, / ;s. 7 --A -L 52C Isla[) to.I 5,7 ,5,A 14.19 5@to os,@A ?5.5? AIR _js,io TEM .5.5 c Z,o ?-1 2-0- I '@'.ONTROL Il pH of Activated Sludge Mfxed Liquor: PH of highest concentration tested: Test Temperature: 2.0 -.ZZOC, Initial lo-to Initial(p.(p Finallo.9 Final (p.15 EWIRONmE.NrAL 1AWRATORY ACTIVATED SUJDGE RESPIRATION INIIBITION TESR #209 . CALCULATION OF RESULTS Te st Substance-I-yc-mirF- 30 MIMM EXPOSLIPX- Test Substance Concentration (miz/L) CnNmnl - T 'IS20 tilan Oxygen Uptake Rate R o.2 4tir7 18 R Corrected(') to.2e C 1. ---I,L) LR I-IIAIRO -/ Date .VJ,9loA Analyst -JZ4Z pag e-z 6-ris I Inhibitionin Respiration Rate(2) CON7ROL II (3) 4Percent Differencein Controls Bcso (95% Confidence Limits) mg/L -A. z I IIILZP(IC;IRP___ Test Substance Concentration(mg/L) CONTROL I Oxygen Uptake Rate R o. RC)@(Q -1'7-ZA I "IO-Ci 0,14q I R Corrected(') to 2(fC t Inhil)itiion RespirationRate(2) I CONML II PercentDifferencein Controls(3)-. L'C-so(gs% Confidence Limits) mg/L (1) Temp. CorrectiLon-- (mg 02/L-min.) RC 1 + RC2@ .. 2 RS (2) 1 Inhibition-- x 100 1 + RC2) Where: (LC 2 / RToo-bsOxy-gSeanmupplteatkeemrpa.t,e*C(mg 02/L-min.) & - 1.047, x - T,obs - 2eC C n Control I_RC (3) % Difference x lo@ I + RC2\ 2 The two control respiration rates should be within 1S% of each other 19 41 ug K to 10, cr car E 4 v + +1 41 a 0 c- oc V, 10 LOI c ec v., Uac9n I x CL LD Eco E .2 it CL L 'orm IOUO -8 -D -PWO Sample Description -A v-Ci@lQRCA Label Dvl)ow-0 STOCK SOUITION: Environmental Lab - Work Shoot OECD ACRIVATED SUMGE RESPIRikTION INHIBITION TEST #209 Lab Request No. Page of Date: --,/1/9 190 Analyst: Stock and Test Solutions Preparation -,IC-JIPt-I b-,+vaicc:iar@ L-.E6@"14,5t-o (p IML, =n.r--imQ k=r4 -11, le) mt, Test Conc. ml stock soln mg/L per SOO ml ml M:) water per SOO mi 4. Bl@ cuaijml m 0 ml syn. sewage feed per SOO ml ml microbial inoculum per SOO ml lip Lo() + Total volume ml osoo- r--@loknktim i4" W)4 ENVIRONMENTALLABORATORY LRY4 /@'dgeZ07@5 OECD ACTIVATED SLUDGE RESPIRATION INIIIBITION TEST 0209 RAW DATA Test Substance: --2S5, Source of Activat;d Sludge Mixe'dLiquor:g@= DatReeceived.:1IIrqilie) Dc f Analyst SLh-,-4nkICe-) kne;- Date Used: ni Iq 190 Ij M14, 30 - MINUTE EXPOSURE Test 5ubStance Concentration(mg/L) 0.5 CONTROL IL DISSOLVED.OXYGEN CONCE TRAIIONS--ma/L TIM INUT)S TEMP 1.0 11.5 2-.-0 2.5 13.0 3. 4.0 14.5 5.0 5--S rIc 1 4 -q --z6i. ;!k s.o 4.%l 44@.44 ZP2Tr'I'3cM1' CONTROL M 6-o 3 - HOUR EXPOSURE lest Substance Concentration(mg/L) 0.5 CONTROL rl DISSOLVED OXYGEN COMCENTRATIONS mall IME- INUTES IFTEECMMPI 1.0 11.5 2.0 2.5 13.0 13.5 14.0 4.5 J5.0 @-5..55.@5 OC R..:; 1-05.1 4.lo 4.L zl.q L'Il;?1.1 ID.q 6,1111 [0.10 6.4 "ONTROL M L4. L4.1 7 pH of Activated Sludge Mixed Liquor: pH of highest concentrationtested: Test Temperature:Zl- Z40(:, Initialto-to Initial(P.1 ?I Final(o.,g Final6.8 li.o ACRIVATED Test Substance -K- rj'iVIMJI)MLiNiJkL IAABUKAUII(Y SUJDGE RESPIRATION INlilBITION TEST CALLWTION OF RE-SULTS #209 30 - MINURE EXPOSupX- Test Substance Concentration (mq/.L) Oxygen Uptake Rate R t).-Rl,,JSQ 2 R Corrected(') to 2e C -- n@"-islot LR -14-litgO Date Analyst 0 t Inhibitionin RespirationRate-(2) OOMMOL M 10, Percent Differencein Controls(3) Bcs,o(9SI ConfidenceLimits) mg/L Test Substance CI o..n.c.e.wn.-.t.r."ation(m-g-/L) CONMOL IM Oxygen Uptake Rate - R n.-AI442, R Corrected(') to 2dc t Inhil)itiion (2) Respiration Rate S14l CONML (),q(),S5 Percent Difference in Controls(3) ECSO (9st Confidence Limits) mg/L RT @;s (1)Tev. Correction= (mg 02/L-min.) 1 + RC2@ (2)t Inhibition 2 RS - x 100 RC1 + RC2) lqhere: 2 RT,o-bOsx-ygSeanmupplteatkeemrpa.t,e*c(mg 02/L-min.) & - 1.047, x - T,obs - 2(fC C - Control IRC (3) Difference- 1 --9ilxob (RC, + RCN2 7 2 The two control respirationrates should be within 1S% of each other