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: TOXICITY TO AQUATIC PLANTS AR R86 -- O44 EST SUBSTANCE Identity: A mixture containing perfluorooctanesulfonate; may also be referred to as PFOS, FC-95, or as a component of FC-203C and L-5440. (1-Octanesulfonic acid, 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8-heptadecafluoro-, potassium salt, CAS # 2795-39-3) Remarks: The 3M production lot number was not noted. The test sample is FC-203C and is referred to by the testing laboratory as "81-1/8". Current information indicates it is a mixture of 1.47% test substance, 60% `water, 30% diethylene glycol butyl ether, 3% sodium octyl sulfate, 3% hydroxy foamer, 10% polyoxyethylene monooctylphenyl ether, 0.12% sodium lauryl sulfate, and 0.05% benzotriazol. - - The following summary applies to thetest sampleas a mixtureofthe test substance in watersolution with incompletely characterized `concentrationof impurities. Data may not accurately relate toxicity of the test sample with that of the fluorochemical component. METHOD: Method: USEPA-- 600/9-78-018 and ASTM-E-35.21 Draft No. 1. Type (test type): Acute Static GLP: No Year study performed: 1981 Species: Selenastrum capricomutum Source: Originally obtained from USEPA-ERL, Corvallis, Oregon, maintained in culture medium at the 3M Environmental Laboratory, St. Paul, MN. Element basis: Algal cell counts (cells/ml), cell dry weights. Exposure period: 96 hours Statistical Methods: Regression model. Analytical monitoring: pH and temperature. Test Conditions: Algal Nutrient Medium: Nutrient medium per EPA -- 600/9-78-018. This nutrient served as the diluent for all algal operations. The pHof this synthetic algal medium was adjusted to 7.5 prior fo use in assays. Stock and test solution preparation: Stocksolutionswere prepared in the algal standard test medium, however, no further description was provided. 1038 Exposure vessels: 250 ml Erlenmeyers containing 50 mi test solution. Agitation: Shaken continuously at 100 pm. Number of replicates: 3 INnuitmiablecrelolflcooandicnegn:tr1a.t0ixon1s0: fcievlelsp/lmuLs. a blank control Lighting: 400 foot-candles from continuous cool-white fluorescent Wliagthteirng.Chemistry: pH range (0-96 hours): Not noted. Test temperature range (0-96 hours): 23 + 2C (incubator) RESULTS Nominal concentrations: Bk control, 280, 420, 650, 1,000, and 1,550 mg/L. - Element values: 9966--hhoouurr EECC5500 ACleglall-DCreyllweCioguhntt:: 440280 ((119576 ---- 986985)) mmgg//LL. Element values are based on nominal concentrations. Reversibility of Growth Inhibition: Aliquots of the 1,000 and 1,550 mg/L test solutions were diluted with algal medium and cultured for 10 days. Based on growth observed in the recovery phase, the effect on algal growth was found to be algistatic. Rdeesmcarrikbesd: iTnetshteintgeswtassubcsotnadnucceterdemoanrktshefimelidx.tuTrheoefvtahleuetsesrtespuorbtsetdanacpeplays to that mixture and not the fluorochemical components alone. CONCLUSIONS 8T6h8e mtegs/tLs)uabnsdtaanc9e6-ehxohuibritEsCaso9b6-ahsoeudroEnCsceollcedlrlycwoeuingthvtaolfu4e0o8f 4(201(--15999765--) mL. This test substance was determined to be algistatic. Submitter: 3M Company, Environmental Laboratory, P.O. Box 33331, St. Paul, Minnesota, 55133 DATA QUALITY 001029 Reliability: Kiimisch ranking = 2. This study meets the criteria for quality tseusbtsintga.ncHeoawenvdeirt ,latchkessatnuadlyytilcaaclkscoinnffiorrmmaattiioonnoofnthpeuriatmyoofutnhteoftest fluorochemical proportion in the solution. There are also some errors (presumed typographical errors) in the report. The Technical Report cover sheet indicates a "40 day exposure" and page 2 indicates the algal culture was stored in the dark "at 40C". REFERENCES Testwas conducted by the 3M Company, Environmental Laboratory, St. Paul, MN, Lab Request number 6811S, 1981. Dm TH eeammm---------------- Last changed: 6/26/00 on1020 i wife Y yk Fa TECHNICAL REPORT SUMMARY oi recntigal coumumcaTiow cei 201201 {importan~tIf reipsprointredotnbothsiodfpaepesr,sendtwocopietso TCC.) - 6/12/81 Environmental Engineering & Pollution Control (Envir. Laboratory) 0535 Commercial Chemicals 9970012600 Algal Assay: Bottle Test(AA:BT), Toxicity Testing FC-203C 050 E. A. Reiner - 21-2 LR #6811 M. T'. Elnabarawy - 2-3E 46981 - ecomrvp |B oon 7 O Glows SacHEMeAL 5 enShin gris emonos, CURRENT ORIECTIVE: = Sp es y Spots | Tao40d-edtaeyrmienxepostuhreealpgearliodgrotwothvarryeisnpgonsceosnce(Cngt,@tfo)nsfoloflowFiCn-g2 followed by a 10-day recovery period. EE & PC Div. (Env. Lab) tga Assay TBoettt aie, v ONTARTA,(250250 wna ois rv orion hSw 7 To Gon The algal acute 4-day ECg, values obtained for freshwater gtroeeFnC-a2l0g3aCein(tSreliepnlaisctartume'Beaxppreircimoernntustwuem)refo4l0l8owianndg. e42x0pomsgu/rle These values represent the algal growth response as measur by cell-dry weight and algal cell-count, respectively. No test material concentration was defined as the no-effec level. `The algal biomass monitoring measurements following: a 10-d: rceocnocveenrtyrapteiroinosdteisntdeidc.at.edSinicnesitghneifiaclagnatl acellgliscidhaalveefrfeecxotvear and resumed logarithmic growth when resuspended in fresh medmium, the 4-day median growth response (ECgq) is . interpreted as an inhibitory effect. BEST OPY AVAILABLE . Ce.3 Cn " oan di a a Lo aSRE SeenLe Lnee Kooi oaRN E SE S ie ST #8 , Sher, t nseim NanRe on5 Od|| 2 Lp ERR SR BR alCleShlhotaati nT ge CAR rma ve ONO] - FPCag-e203TiCo/MTE June 23, 1981 concruszons In these triplicated 4-day ECsg algal assays, the calculated rwveearslepuoens4s0e8foaarsndfmre4ea2s0shuwrmaegtd/elr.bygrTecheeenlsle-aalvrgayaleuwee(siSgerhletepnraaenssdternuatmlgaClatphe creaillclgorancluoutgnurtmos)wth respectively. No test material concentration was defined as the. no-effect level. The algal biomass monitoring measurements following a 10-day recovery period indicated little or no algicidal effects at concentrations tested. Since the algal cells have recovered -and - resumed logarithmic 4-day median growth grreoswptohnsewhe(nECsrqe)susipse-nidnetderpirnetferdeshasmeadnium, the inhibitory effect. INTRODUCTION In testing the possible effects of chemical substances on the aquatic environment, unicellular algae are recommended as a model system for evaluating the influence of chemicals on aquatic primary producers, plants, and phytoplankton. Toxic effects are tested on growing algal cultures that undergo cell-division during the test. Algal growth response include: a) no effect, b) stimulatory, c) inhibitory, d) algistatic, or algicidal. Tahmebietnetstrosoumbsttaenmcpeeraitsurae.cleaTrhesaembbeiroalsisqauyisd, soluble performed in on water this at material evaluated generated form the its potential basis of this algal toxicity report. and the data MATERIAL AND EXPERIMENTAL DESIGN These algal assays consisted of a 4-day exposure period followed by a 10-day recovery period. The test protocol utilized for this study was modified after those described by USEPA - 600/9-78-018 and ASTM-E-35.21 Draft No. 1. (1), (2). Test Species: A culture of the freshwater green algae (Chlorophyceze), oSebltenaaisntreFudrmomCaDpErEicPoArnuEtRuEm, ofCotrhve., orOdreergonchl(oFreobcroucacrayle1s3, wa1s981). The test algae are non-motile unicellular cells and having the appearance of a new moon. The algal culture was stored in the dark at 40C until use. dmeirE sL ualE eliE E a CAEL C tATgEr a eotolae FPaCg-e20T3hr/eMeTE June 23, 1981 Inoculum: gAilvgeaea fsrtoamrtain7g-daoyptiomldumsitnocokculcuumltulreevelwerofe gurseedenaaslgianeoculum to 7S-e1l0ednaasytroludm cCaupsrtolickctorunurtuem 1wixll104incseulrles/tmhle. preTsheencueseofofaa sgruofwftihciephnatsea.mounTtheofinviitiaablleaalglaglalceclelllscouinnt thien etxheponsetnotcikal C(u3l7t0u,r0e00wacselldse/tmelr)m.ined with the aid of a hemocytometer Algal Culture Medium: . ASttatnadcahrmdentcul#t1.ureThainsd atlegsatlmecduiltuumrewasmedpiruemparperdoviadsedoutalllined in @niultureinetntsforessalelnfiaallgalfoorpearlagtailongsrowitnhcluadnidngalstohe sperrveepdaraatsiotnheof aSdtjoucsktesdoluttoio7n.s5. priTohre ptoH oufse thinisasssyanytsh.etic algal medium was Culture Flasks: - Ecaocmhpri2s5e0dmla etersltenmfelyasekr. conCtualitnuirneg fl5a0skmsl oafndtealslt ostohleurtion g#l2.asswAaurteoclwaevreed spfeocamiaplllyugsprewperaereduseads adsescfrliabsekdclionsuArtetsa,chment allowing for free gas exchange. Range-Finding Test (Exploratory): bThieomaesxsplionratsoirnygletesftlasckosnsicsotnetdainoifngdetsetramnidnairndg ctuhletur4e-dmayediaulmgal porlduesrstheoftmeasgtnimtautdeer:ial0.a1t, co1.n0c,ent10r,ati1o0n0sancdove1r,i0n0g0 msge/vle.ral Full-Scale Test(Definitive): Aulslingdef2i5n0itmliveErlaelgnamleyearssafylsaskwsereconctaarirniiendgou5t0 miln otfriptlesitcate sstoalguetsi:on.a T4h-edaydeefxipnoistuirvee pteersitodconfsoilsltoewdedofbytawo1c0u-dlatyure preercioovderywaspetroiodv.erifTyheipnuhripboisteoryofeftfheectesx.posuTrhee r(eccoonvteacrty) a(lsguibscutlattuirce/)algsitcaigdealproevfifedcetds.information on ~~ T65h0e, f1o,l0l0o0winagndFC1-,255003Cmgl/olgarweirtehmiucsedcontcoenitnriattiiaotne: th2e80,4-d4a20y, cbornatcakcett p4e-rdiaoyd.ECs `Thviasludeosseprreandgiecteodf dfriolmutiporneslimwiansarsyelected to ealxgpalloractuolrtuyretesmteidnigu.m plTuhsreealgfallaskcsellcsontcaoimnpirnigsed100t8hefresh nontreated controls. The recovery stage was initiated by CatU anRaoRRGealB AREY a See ks REAR FL ES 001043 FPCag-e203FoCu/rM,TE June 23, 1981 - . osfubcthueltutrwionghig(h0e.s5tm1t/es5t0 cmlo)ncefnrtormateiaocnhsofinttohefltahsrkese croenptlaiicnaitnegs effrfeeschtsalggarleatmeerdiutmh.an T8h0e%sedurtiwnogttrheeatmcoennttasctinpdeurcieodd.inhibitory tFhreeshalgsatlockstasnodlaurtdiontsestofmetdhieumt.estSeseubAstttaanccehmewnetre #3p.repared in Test Conditions: cAlolndiatligoanls,opeinratoirodnesr twoereavociadrriceodntaomuitnautnidoenr swittehrilbeacteria and eontvhierronamlegnate.al cAlhgaamlberculutnudreersthweerefolilnocwuibnagtedstainndaarnd growth . conditions: *Temperature - 23 + 2C (70-779F) "Fluorescent illumination - 400 &.C. + 10% *Sfhreaekinggas 10e0xc+han1g0e rp-m.Continuous Platform - Algal Biomass Monitoring: wTheeighatlga(lmgg/1r)owtahnd rbeyspcoenlsle cwaosuntmea(scuerlelds/mbly).algaBlothceglrlo-wdtrhy meexapsosuurreemenptesriowdereandmadoeilywitche.lla-lcgaoluntculwtasuremsadeafwtietrh tthhee 4-day osfulbciuolmtausrsesmeaafstuerremaen1t0s-daarye rbercioevfelryy dpeesricordi.bedBionthAptrtaocchemdeunrtes . uBtCislgizvianlgues34anSdIKC9U5R4, coanfTiRAdCencSeystleimnitfsorwerreegrecsaslicounlatmeoddels of experimental data. TEST RESULTS TSheelenraessturlutms oCfapbriiocmoarsnsutumemasuuserdemeinntstheofse tshetudgireesenaraelgae Sreesullfts-eofxpthle adneafsnidtnaiotrirevyedteetsatileadre isnumdmaatraizsehdeetisn Taatbtlaecheld.. The Et hare RREHICSE 00100eli FPacg-e203Fciv/euTe June 23, 1981 : Table 1 Definitive Test - ECsg values, mg/l with Selenastrum Capricornutum --Typ-- e Tes-- t --Cel-- l-Dry-- Weigh-- tIGmgT/-- l bio-- meCe11-- FCoonuenrt-- No./-- 1 mL Ferioa 4-Day Exposure 408(156-995)* 420(197-868)* #954 Confidence limifs. * The data reported herein are based on studies performed by M. T. Elnabarawy. Accompanying data sheets comprise original data. A copy of this report is filed in the Environmental Laboratory Archive System, Building 2-3E. It should b noted that the reported ECsg values were evaluated RerERENCES oinn, ttheestbassoilsutiofonst.he calculated initial concentrations of FC-203C (1) Miller, W. E., J. C. Greene, and T. Shiroyama. 1978. The Selenastrum Capricornutum Printz Algal Assay Bottle Test: UE.xSp.eriEmnevnitraolnmdeenstiagln,PraoptpelcitciaotnioAng,encayn,d Cdaotravalilnitse,rprOerteagtoino.n. EPA-600/9-78-018. 125 p. (2) CAoSnTdMu-cEt-i3n5g.2iT.oxic1i9t80y. TesPtrsopwoistehd Standard Practice For Freshwater and Saltwater Algae. Draft No. 1. ikLBEa Ge Sa TARE e 0010i45 oH : TA bieA Laban i rafoaLsaelCe CRE : Type Test 4 Day Simmer sonar ALGALTOAXSISCAIYT:Y TBEOSTTTLEOATTAESTSHEE(ATA:BT) Eqpos ce w_G38lilg Material Tested_EC - 203 C Lot Physical Description Clear Amboy Lig xd Algal Species:Green TEAT lgse, Selenasitnruenccaaprrliccoornnuutninm source Corvallis | Oregen Date Rec'd 2-8-7) Age of Inoculon__ 7 days Test Container:250 ml Erlenmeyer flasks with fomn plugs Sample-to-Volune Ratio:__SO ts 250 initial Call Loading: 10000 cells/ml Initial pH of Synthetic Algal Medium: 7,520.1 Test Conditions: TFeimepterersacteunrte =11T2a3T%2eatie(70o-77tF tee g10% . Free Gas Exchange -Continuousplatform-shakingat 100210rpm. Date Started G- | - 2 Date Completed Go -[S-F Investigator MT, E lnabgSo ALGAL CELL DRY WEIGHTI, MEAN VALUE inmg/1 (1) TREATHENT EXPOSURE: 4-DAY CONTACT PERIOD RECOVERY: 10-DAY SUBCULTURE PER100 oNTRO $0 ee 6.5 i. 5.T 420. 1. 2.4 --~la2S000 LEO oe tdriam nsanirrronentri-er -G resets mi------------ oe revs sevefl eee ie its ---- ss spell mmmsictnpommmondine agarose ger 0 -- ALGAL te CELLCOUNT,MEANVALUENO,/mi{1) LONTRO 20 o eso --L.000. 50 gC RET 400.000 0,000 69,000 1 PY) rednBDe OO.e .. 0,000 ERA le Creer ee | 0,000 innlO,000 00,000 Aico (1) (2) Based on mean values of Method of calculations; each triplicate sets of flasks. BEST COPY AVAILABLE BE Go Terr of == coe oom 001015 TREATMENT ALGALEANSVSIARYO:NMEBNOTTATLLE LTAEBSOTRAT(OARAY:BT) , RECOVERY STAGE DATA SUMMARY ALGAL CELL COUNT, Cells/ml (MEAN VALUES) w_G8 Ss -- oO ~< INITIAL ALGAL CELL LOADING a FINAL ALGAL CELL COUNT ' veel 700 190m 2 sso] se~ e [ioosso I J] Ea (1). Based on 0.5m1/50m! subculture made from k-day exposure cultures. (2). 10-day recovery cultures. Cae a REE 001047