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ACTIVATED SLUDGE RESPIRATION INHIBITION TEST TEST SUBSTANCE Identity:A mixturecontainingperfluorooctanesulfonatweh,ich may alsobe referredtoas PFOS, FC-95, or as a component of FC-206CE. (IOctanesulfoniaccid)(CAS # 2795-39-3). Remarks: The 3M productionlotnumber was 502. The testsample isFC-206CE. Currentinformationindicatesitisa mixtureof 0.92% PFOS, 15% diethyleneglycolbutylether,76.09% water,4% urea, 1.28% Sultonefoamer,1.55% sodium octylsulfate,1.05% polyoxyethylene monooctylphenylether,0.06% sodium laurylsulfatea,nd 0.05% tolytriazole. The followingsummary applies to a mixture with incompletely characterizedconcentrations of impuilties.Data may not accurately retlectthe toxicityof the fluorochemical component of the testsample. METHOD Method: OECD 209 Test type: Acute static GLP: No Year Completed: 1989 Analyticalmonitoring: Dissolvedoxygen concentrationpsH,, and temp. Statisticamlethods: Resultswere determinedby calculationfthe% inhibitioin 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 testsolutionpreparation: A stocksolutionofthe referencesubstance,3,5-dichlorophenowla,s prepared by dissolving 500 mg inlOmL IN NAOH, dilutedto 30mL withMilliporMeilli-QTM water,then broughttothe pointofincipienptrecipitatiwointh1N H2SO4 and dilutedto 1L withMilli-QTwmater. The pH ofthereferencesolution was measured to be 7.6.Test substance was added direcuytothe testvesselsand was apparentlymisciblewithwater. Test vessels: Not given. Number of concentrations: fiveplus 1 referencesubstance and 2 blank controls Temperature: 20 - 22 *C Totalsuspended solidsand pH on day of testing:3.6 g/L(TSS), initiaplH 7.3and finalpH 7.3 Element Basis: Respirationinhibitioans determined by oxygen consumption. Remarks: Valuescorrectetdo20 "Cforcalculations. RESULTS Nominal concentrations: 2 Blank controlsr,eferencesubstance,100,180, 320, 560, and 1000 mg/L Element values: 30-minute EC5o = >1000 mg/L 3-hour EC5o = >1000 mg/L Element valuesbased on nominal concentrations. Remarks: Testingwas conducted on a mixtureas describedintheTest Substance Remarks field.The values reportedapplyto thatmixtureand not the fluorochemicalproportionalone. CONCLUSIONS The FC-206CE 3-hourEC50 foractivatedsludgerespiratioinnhibitiownas determined to be >1000 mg/L. Submifter: 3M Company, Environmental Laboratory,P.O. Box 33331, St. Paul,Minnesota,55133 DATA QUALITY ReliabilityK:limischranking2. Testingmeets allcriterifaorqualitytesting. However, sample puritywas notproperlycharacterizedand the studylacks analyticaclonfirmationoftheamount offluorochemicalproportioninthe solution. REFERENCES This studywas conducted by the3M Company, EnvironmentalLaboratory,St Paul,MN, Lab Request number Gl 358-2,1989. OTHER Last changed: 6/26/00 ENVIRONMENTAL LABORATORY ACTIVATED SLUDGE MIXED LIQUOR DateAnalyii"IcAi Type Teat 0 &C! t@l-AC41*uaiSelcitx)cie. \.i I :rvtbi'bi'-+fe)-o@*m#-17cq Source Date Obtained T-reLt+rheiI*lidyi4 Date Used Mixed Liquor Suspended Solids (MLSS) 3 to g/L pH ,T'V1I'J4:-Ia = -7, F#hj t '7.9 Pqu/-@ A-,y :3 AdJustment_A,,Ovie g/L Coments Te&+- ry" u2i4.14-,-I-P- & 17, Test Substance Sample Description_ FC - &qO& er- I-e)+ Samplepl@-,qjqhe-4(on1e0.000 /1-t.7..@Adjustme1n4tlotle Comments Reference Substance Sample DescriptiorL Sample pH 7.k Coments /6),CCL <nth.\ Adjustment Exposure Conditions TestDuration Test Temperature_ o - Oc Comments Form IOM-8- 0- PM Sample Description ,=C-Ao6e-E L c i 5C),P,- EnvlronmentLaalb- WorkShoot OECD ACRIVAUM SLUDGE RESPIRATION INHIBITION TEST #209 Lab RequetiNto.6 1,53,9 Page I of 3 Date: to-,93-91 Analy.t:,A.@""q - &-a@ Tt-qtgnliitinn-P,-reparation Test Conc. mg added iniy/T.. r--r @00 mi ml bf-water ml syn. sewage ner SOO ml fped npr SOn mi ml microbial i]3nriilinvenE @g()Oml 1 C)C> INC> So + qc > + + Ito 4,@o + IL + 4 ac>C) 4- goo .3,;o@ '571C0) 1000 1100 4 olgc) 4. 6q gq ,@00 j- 01 4- /6 /6 4- -I- aloo 4- a C)o 4- Total volume ml 5-c@o 5-oz) B)Ovlk O-on*roi W 1'+Ln L 0, 17@,5-0-c.L)4> Sldylk Onnfrn I II r 7, I) a u-ua@w u5ea -Rfe- Ra recl O.Q ra4i OV@, G@o r+,ect 40-.Qc LR ENVIRONMENTAL LABORATORY OECD ACTIVATED SLUDGE RESPIRATION INHIBITION TEST #209 P6 RAW DATA Test Substance: @F Source of Activated Sludge Mixed Liquor: M e-+ro (I ) aLc,+ e- 'Tie-a-i rre IQ a vi Date Recei d: Analyst: F---- Date Used: 0 30 - MINUTE EXPOSURE - DISSOLVED OXYGEN CONCENTRATIONS ma/L lest 5ubstance TIME-MINUTES TEMF Concentration (mg/L) 0.5 1.0 2.0 13.0 4.0 1 5.0 6.0 1 7.0 8.0 19.0 10.0 C* CONTROL 1 IOD 71 7, L f.-L I.5-1q.D 3.1>",3. 9- 5.S 5-3 1q--Sq--33,913.3:2 7,,6-7. 0 6 -q @5.9-50 3 --7 3,;, ;Z-7 ;?o 7.7 7.q -5-C)q,q 3,9 3,;Z.2bLq.o 7.q 7,,) 5-9 5.1 l@,5r3,93-1 /000 q 19.3 -4 3.1 o@iil1-7 o-(I- CONTROL 11 7,-3 7. 5,0 3.5-130, 3 - HOUR EXPOSURE lest Substance Concentration (mg/L) CONTROL 1 /oo /90 ,3&@cR,) /ooo 0.5 1.0 1-1 7,b 7.X DISSOLVED OXYGEtl COTICENTRATIONS ma/L TIME-MINUTES 2.0 1 3.0 4.0 15.0 6.0 7.0 1 8.0 9.0 k -711,.,1,5, b 15-) Li- L 3-D 'TEM 10.0 c :9c.) '7e,ll7.5 7,, 6.2 D Y.b P6 9.1 7.7 7,1-17.0 @;-1 7,-7 7.3 &,9 7,4,@@ #-9 oail1 '7..-75.'1/,0't.oq.ito,O5.7 1519 q.q 3.0 7-9 7.1-57.1 6.,3 .5-5-5.1 q.5 'OtN'TROL II r7.3 17.1-ib.5'16,o5. 0 pH of Activated Sludge.Mixed Liquor: pH of highest concentration tested: Test Temperature:20-;IOOC Initial7.3 Initial7,5- Y@3,q 3.3 Final 7,3 Final7. 6-- EWIRONC.NFAL LABORNFORY Test Substance A=IVATM SWDGE 01, (1 RESPIRATION INHIBITION TEST #209 CALCUIATION OF RESULTS I-o4-50A 30 - MINLrfE EXPOSUPE. Test Substance Concentration(miz/L) MNM()L T /oo Oxygen Uptake Rate R ---Ol R Corrected(') to 2e C --o Li-1 q776 loon (r-,OOZ) lob /,@? 0. -7//&R --e) .,5731 0- 1 0 -7-1- '7 CDNWL 11 0 ,Percent Differencein Controls(3) *7 1 o -Il(o7-7 1.,5C-;lo ECSO(951ConfidenLciemitsm)g/L-> ]IO)Q-- 1 -(7ADn@aatlyst PA 13, Now I Inhibitionin ResDi ation Rate(2) - Test Substance Concentration(mg/L) CONTROL 1 lc>Q -/Ooo Oxygen Uptake Rate R 0 0 q,50t0 -7-7 00 C') 3,57(o 0 R Corrected(') to 2e C q7q q qal00 C)- 3 77 t, y 000 3@@76 t Inhibitionin Respiration Rate (2) - 13 17 'A h CONTROLil PercentDifferencein Controls(3)-- 0. Z 916 ECSO (95% ConfidenceLimits)mg/L- @ 1C)t)c) MA IL (1) Te-np.Correctionz e (mg 02/L-min.) RC 1 + RC?) (2) 1 Inhibitionltiere: 2 RS RC + RC - 1 2) 2 ,0/ x 100 R - Oxygen uptake rate (mg 02/L-min.) T,obs - Sample temp.,CIO 1.047, x - T,obs C Control IRC (3) Difference- x 100 RC1 + RC2 "2 2 The two controlrespirationrates shouldbe within 151 of each other See, r +es,.-Lcotti. p F- Sample Description 3,5- t>@c6 loroj>heno I A e@e ren ce- 5obs4a nce POA L&6a, Aof c ry4,@a I@ Lab Request No. EnvironmentalLab - Work Sheet Page ot OECD ACTIVATED SLUDM RESPIRATION INHIBITION ITM #209 Date: Analyst: 5-@ Stock and ation 7,!5"0 STOCK SOLUTION: 00 ni - u4ecL *1 rme @A OA) Xb e-) Fest C M,g/L m Der 00 ml water 1 syn. sewage per 500 ml-----ifeepder 500 mi ml mic bia inoculum Der SOO ml Tota volume mi 7 L/ + I)C A4i)lj-- Lk) LR ENVIRONMENTAL LABORATORY OECD ACTIVATED SLUDGE RESPIRATION INHIBITION TEST #209 p@A RAW DATA Test Substance Suk<4d-nc%e_ Source of Activated Sludge Mixed Liquor:A4e+rc> IL)a<.+e-'-Tfea4mpn+f>14n4 -.SI-PaLjl,A4,0 Date Recei ed . to-o?-A Date Used: &-07..g-sq Analystn:d..t, a 30 - MINUTE EXPOSURE - DISSOLVED OXYGEN CgUCENIRATInNS mall iest bubstance TIME-MINUTES TEM Concentration(mg/L) 0.5 '1.0 2.0 13.0 4.0 1 5.0 6.0 1 7.0 8.0 9.0 10. c CONTROL I /D '7. 7.a CONTROL11 7,3 .;@F5.7T,5.i 3 - HOUR EXPOSURE lest 5ubstance Concentration(mg/L) 0.5 CONTROL 1 7.3 DISSOLVED OXYGEN CONCENTRATIONS mall TIME-MINUTES TEUF 1.0 2.0 13.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0 c 7.) 6.5-16.0 q.q ;?-Cl @Q 7.77.5- 7-0 '%'.ONTROLII 17#q 17* 1 b b I pH of Activated Sludge Mixed Liquor: pH of highest concentration tested: Test Temperature:AI-A;(OC Initial7, Initial 7.6@ Final 7.s- Final .7,9 Test Substance ACTIVATED EWII@ONMI'MAL LABMTORY SUJDGE RESPIRATION INHIBITION TEST #209 CALCUIATION OF RESULTS 30 - MINUTE EXpOSUpE Test Substance Concentration- L MNTgni, Oxygen Uptake Rate R R Corrected(') to 2e C O.q&77 & 1.1.58-,R LR.6@@ Date Rq Analysf@ n-ea p@j3&fi I Inhibition-in Respiration Rate(2) CDNML 11 0, 5901 Ll O.,qe? #p Percent Difference in Controls(3)_ ,57, Ecso (95% Confidence Limits)mg/L /0 -1- igLExpngimp /Z- u 1)(ep&- q3 Test Substance Concentr.ad tion (mg/L) - CQQQL I Oxygen Uptake Rate R C). 5A9 R Corrected(') to 2(fC )4/m I Inhibitionin RespirationRate(2) -57 COICWL II 97ql- Percent Difference in (3) Controls 0 v'7 EC so (9S%ConfidencLeimitsm)g/L /z> rnoL-)iIV-%fti)(49A.76c-@7n k Ibl+IC)eu (1) Temp. Correction-- (mg 02/L-min.) + (2) 1 Inhibition 2 R'S x loo RCI + RC2) hhere: 2 R ,10s xIgen uptake rate (mg 0 /L.min.) T Sample temp., C& .2 & - 1.0479 x - T.obs - 2(f-@ C - Control (3) 1 Difference RC - XL;2f .100. RC +@R+C2RC 2 The two c rates should be wlthin 151 of each other See_ p,?,A _Cr.)r c., 4_ te