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AR126-OSIB TOXICITY TO AQUATIC PLANTS (E.G., ALGAE) TEST SUBSTANCE Identity: Perfluorooctanoic acid; may also be referred to as PFOA, FC- 26, or FX-1001. (Octanoic acid, pentadecafluoro-, CAS # 33567-1) Remarks: The 3M production lot number was 269. The test substance is a white powder. The test sample is FC-26 referred to by the test laboratory as N2803-3. The purityof the sample was not sufficiently characterized, although current information indicates it is a mixture of 96.5 100% test substance and 0- 3.5% Cs, C7, and Cs perfluoro homologue compounds. Remarks field: The sample preparation directions given to the laboratory were to dissolve the test material in a 50:50 `water:isopropanol solution. In the protocol amendment, it was stated that the sample was combined with isopropanol in a 50:50 ratio prior to use. METHOD Method: U.S. EPA-TSCA Guideline 797.1050 Test: Static acute GLP: Yes `Year completed: Study completed in 1995. Report completed in 1996. Species: Selenastrum capricormnutum Source: Originally from The Culture Collection of Algae at the University of Texas at Austin, maintained in culture medium at T.R. Wilbury, Inc., Marblehead, MA. Element basis: Reported two ways: number of cells/mL and growth rate. Exposure period: 96-hours Year study conducted: 1995 Year study completed: 1996 Analytical monitoring: Nominal concentrations. pH and temperature. Test organisms laboratory culture: Algae cultures were growing in U.S. EPA-recommended sterile enriched medium for at least 14 days prior to test initiation. Test Conditions: Algal medium: prepared to U.S. EPA recommended concentrations by spiking deionized water with nutrient stocks. The pH of the synthetic algal medium at test initiation was 7.4. Algal medium used for culturing and as diluent. (33936 Ssttoocckksaolnudtitoenstwassolpurteipoanrsepdreinpastreartilieone:nriAch1e,d00m0edmiga/.L Apprpirmoaprryiate amounts of this stock solution were added directly to dilution water to formulate the test Exposure vessels: media. 250 mL glass Erlenmeyer flasks containing 50 mL of test Agitation: solution. Shaken continuously at 100 rpm Number of replicates: three Initial algal Number of cell loading: 1.0 concentrations: X 10 cells/mL Five plus a negative control Water pchHemriasntgrey:: (0-96 hours) 7.4-103 (control exposure) 2.9-3.0 (1,000 mg/L exposure) Test temperature range: (0-96 hours) 235-24.0C Light levels: (0-96 ~380 ft-c from hours) continuous cool-white fluorescent lighting Remarks: The pH of the exposure concentrations test were solutions for in the range the 250, 500 of 2.9-4.0 at and test 1,000 mg/L initiation. This low pH would have adversely affected the survival and subsequent growth of the algae. RESULTS _ _ Nominal concentrations: (Tested as 50:50 mixture of Blank control, 63, 125, 250, 500, test substance and isopropanol). 1,000 The mg/L. concentration of test substance in solution was 32, 63, 130, 250 and 500 mglL. Element value (as-tested): 72-hour ECso (cell density) = 180 (125-250) mg/L 9762--hhoouurr EECCssoo((cgelrlowdetnhsriattye))== 118800((112255--225500))mmgg//LL 96-hour ECso (growth rate) = 180 (125-250) mg/L 96-hour NOEC: 125 mg/L. Element value (based on concentration of test substance in solution): 72-hour ECss (cell density) = 90 (63-130) mg/L 72-hour E.Cso (growth rate) = 90 (63-130) mg/L 9966--hhoouurr EECCsaoo((cgelrlowdetnhsirtayte))==9800((6633--113300))mmgg//LL 96-hour NOEC: 63 mg/L (USE All element values based on nominal concentrations. A toxicity test conducted previously with isopropanol at 1,000 mg/L showed no effect on algal growth. Statistical methods: Cell densities, growth rates and percent inhibition values used to estimate the EC1o, ECs, and EC values and 95% confidence limits were calculated using the computer software of C.E. Stephan. The no observed effect concentration (NOEC) was calculated using one-way analysis of variance (ANOVA). Control response: Satisfactory Biological observations after 96-hours: C(aNionomscienuda,n [o[[fmMteCanaNupemober||vPDieertcneonatvri|a s[GrpohewrtchienRtavnitae | [coma[o|m[e| n w | mon| 2|o | [oo loom | w | wm | w |woow|w | wm | om |wo|mwo o| w | Observations: Algal cell counts in each test vessel were determined by means of direct microscope counts with a hemocytometer. After 96 hours of exposure, there were no signsof aggregation, flocculation or adherence of the algae to the flasks in the control or any test treatment group. In addition, there were no noticeable changes in cell size, color or morphology when compared to the control. Reversibility of Growth Inhibition: Effectofthe test substance was determined to be algistatic based on the results of the post-definitive test exposure. CONCLUSIONS The test sample 96-hour ECso and 95% confidence interval for Selenastrum capricornutum was determined using two calculation methods. The as-tested test substance 96-hour ECs for Selenastrum capricornutum was determined to be 180 mg/L with a 95% confidence interval of 125 -- 260 mg/L, when calculated using either the cell density or CI39Ls growth rate. The as-lested test substance 96-hour no observed effect concentration (NOEC) was 125 mg/L. The 86-hour ECs for Selenastrum capricornutum based on the concentration of test substance in solution was determined to be SO mg/L with a 95% confidence interval of 63 ~ 130 mg/L using either the cell density or growth rate. The 96-hour no observed effect concentration (NOEC) for the test substance in solution was 63 mg/L. No signs of aggregation, flocculation, or adherence were. noted in any of the test solutions. This test substance was determined to be algistatic. `Submitter: 3M Company, Environmental Laboratory, P.O. Box 33331, St. Paul, Minnesota, 55133 DATAQUALITY _ _ Reliability: Kiimisch ranking 3. The study lacks analytical measurement of test substance concentrations in the test solutions and sample purity is not sufficiently characterized. Additionally, there appears to be a discrepancy between the sample preparation directions given to the laboratory and the procedure conducted by the laboratory to prepare the test solutions. Initial low pH values in the higher concentrations may have had an adverse effect on the survival and growth of the algae. REFERENCES This study was conducted at T.R. Wilbury Laboratories, Inc., Marblehead, MA, at the request of the 3M Company. OTHER Last changed: 5/25/00 3E830 comt pts ety ns ere se ien--s--vt--m--e cor SR pres ; `mote ; TLE | orea ein | |! 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Index of Tables and Figures VILE. athods and vatesiate vv . s 4 5: X | ,i .| .! Appendix A. Water Quality Data from Toxicity Test er 7 Zs 3 i Sa a hs LTE ERR Ne S F SodsmReIeES ene LCS losedSR CL SE a Ca Damas) foike cn Fos ER RL ae Hm ag adi ik aE > WI pe Fr SE Se e JR Ch %:% a th SSiEE Shi dn ysi SER 0013 CAE Fle. cHmhinLtsenLTtCS hEsstSmaiL ereraamiaeI oneeofrs4I rtS epsastfay nsivn . Ble 3 oBEeRnaEte,RsTaI ma fo eh totety sos wich ] Foes ony arm te cnet " i Rte 4, wos parI sriogi cohdigtropye waatnmtoeesfSehmmorsnatoatytedsoomnent - i Eermates sie Sosastrin opslesmmseeih he Totes. SgjruEemekTsni pmenn oinosKopihiltotifika ih " Rte 0.1. SeS paTcsEL acEsmLa: ueSed feinnth e totee tae 5 Fe. L ptI enAGAa cotnetcaeeowsvis 0 E BB L i; r L enbe % eaes, ssceh apemridceoernsnutcS urfneoontRE RSSl RERYe Ea BSha ERALdRyR1L 5WkL SShlaielCa el SL C BL TRE HB RA SSR Oy MRC EuiRi T a . ai ASR a bea rE ea a J Re i SE a ETyh LH03941 SRB mre Ch abe les a IsiResisgiMoAS 20 sin BSooe SReSigWeyPolar || aBm Torn aLcaomeounyefTiornot3SshooseTetsennkso)spsJounietrioeTtbaeovrpi1c2t,s enSTsEv,erantweoommnseTiesizeo2ry + tomperatuce of 20t 2. | ! pnThe sample of K2E803D-3 Ssupptlied2by ,the sa ponsor vease ctombigned v3ith |{ Barrie ai ReyESi,RayFEore ried |! EE SiriEirneamaar ! Sri ER eat n SCaR RSNE SE SiaeSneEHsatnkp7,eCS e5gS i 0ErSWEeei h 2gTeev atidpwEiReiate th8 || TE H Ei IDn 5Pi) H LSoeSy baRJot ors || 5, 5s LBpaeedass o2 wRE the average specific growth Kate and the 36 hour NoEC 1a' 63 mg/h and the 1170 | Saopropasal) Ea TE | RIM 1G ASR a sSaadpboe oTSEbAEaRlEataaEg<rLA e SG C RR o S R R N Res Si ASNEIET ET ke SREA igi A LaEPiR C e i POR RT aime i EER ds a OR bard .2 ok en3945 ot Mi TE En i RE" in fue ; Expctmncas compotion i SET Fa TIT Arh eycris, ed PEL Boia a aha a COE LB F i oaaaes l oa e aCSda atimLlfe nS SBEe HERba e E e r eSe CTa ee BERIT voor ones aan a S Si E p ERG OeTTA 0 nFAGNe imBAen E ER E a jp i aFR CR iho Lact io c0ug4s ns ik: IiE BEST COPY AVAIL 4BL a Stahnti3drSvLa pE encbey30cR ompaY.e e he cE e htactiv0oef ttheeaataay S S53 R condiiR oense,ethCe Sohint broa iodal vTaasanbaessecdeonn i0i-5" Eso,Tnhcmatemitses,s,So05eF8 2 Sra 0,018 SL HBOS 5. wibucy Lascatoi cin sateae measeer 531 } Sineeninaealy soi ghee ae II ag bottle included i | i r20i 3 (4 SWhiLte sRolidL) ves wS gpisL d byS I come pany, sv act TIL hn orn sedCREaESE ms SX Teopcooanoty sea potion ELA PL rtalewaded LP S80 eo cr cnt 1 sich ti SEAR pp SEmSrEBe aYL ber R oiencarm go pi T of nGiEaREY Bn eR ee Chern 0a BS TEERRE gnrf incn dl esrle i ceeTn ea ne {setaS nasttruen S clopbiciorcnlie teluns m re Msay STOoAS RE h nDeTeSd eer EeeE Senmtainae irneaatt FEED: BN eriatyorb ticlr e hioSan repens omen oy A BENBS es a a B Su TRo ANS Cr n eb 3osra A La IER Sali ae =; - or - EE B--e-- t -- : Ta z Signa aE w,e i ARis SHIpi LA i Re SaSE GEST COPY AVAILABLE Table 1. S dBOa LA SSE chemical characterisation of a representative sample of test i" --P_a--rameter' fSee oo Im,Phosphorus Nessuresent EoE e oi uals sn a SE Lait waleae a3h3 a i:- 0.03 ok he 7.4 te ous Wis Be I fe oa Masntc aan Se n o oat wd ERE isa SEos EBEe oo 2 EErono IoBEna oo Be I03.z0)3 0EB .0a8 a a ivy eo on 2 md rt are RPE ius TT oxsghese Iaasni soz wo da 2os vw or ie EE pr I. . rene. LT ba 0.5 XRAY: E ggg a e re o acad ox G0 eEAE onS IRRekA iCT hm et A wE e e 33 Ft sala Seo en ct "E atel s Smr a SDBRn AOE RT LSAAR abA eyR sty masenr aLE SESS a ER Ee E an eel : : H Eein Fhe i : 4 ; Nd PARR Ee = oi (0394 CH oslo BESTOOPY AVI 131 BREIiT sSUITING ms ae nel a1 SO j SL Ean el wiems ; BE voy i Sa Be SE lin 1 commie 5 SICILY Sr mn e201 wn ive BATL re Te le |! 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Cutcutacas Ung the abr of cottaret 2 Culculaed Unig the aeess spacitic Groh nce CThlooloitnets based co Tub mbsses aessceived (R510 sithent A. Cutcuacas Ung the mmr of cttarit 5. Cheutatas ato th Mace sail sem ace : EneSia Ie Eee TRESS sei Ges 00 wats cote on crevasse hy CIc TESEe ER c L ee BR Late a a Tdi EAhrRSa E ae " R EE SsOer a in ibe ses) er rac la Rea pee he eh it is SfSE en aE aE an ga h EE o LERaRGRe R:b| BSEfNE A : oh Re Ws C05 RE SR ER ERR Ne BEST COPY AVAN Apii Y ; Bre,1Sinndwre rsL3Asistent ra esi Lo iting phan, C5. 1903 Comptes rep fr Coeutation of 165 vain, 0.5 1an0E ameAE untacrsS cpicn omti rene iee snem pte CO iantioens rinek melonseevionpiperAS CONT Act Tene Stn don 10 rte nt sas cont es vers th T2o5xi1cit5y0aatai,thn1d13453 pseende,tha9Fe.ostcmesoanhasined, e"pShacuetiod re ena or mi Ep Company. iby Tete ad LoS nd A idhsa = Te we plaoaans Sn Bi. RR EShaLaE GL iC H neas m a RE i se Te e eeS E RE e ERAEETREa B Re SEa R 2 dR y van mn fCe lE mr iiL einaE la ehl on B ohms RIG a SAE ria ca ey i] 03953 ato is | aR Sor ARE 2 HE SL 3 BEST COPY AVAIL AB n 1 i: OTfifea5Vkoge fut, | uA lint: 1344 : Fi Engrabin%Yoder 29960 3' os TCEHAReNE als, HL fR osts ers SERS 14 LEE ne Shik al bE SAeR LOlE Nac ER e oneLiF cFoemueeehty l ore"+. TT oSurEiI iBbpeReMeInStNEE GCSoRI LEFopL ekt T J EE Pr cone EE Bk [ fo LAE a : Sd tagass oh i CELE Gag p eflEAbR ao peo R ee R i BESTCOPY 11, EY 23 11. en,waesss sn Din proc vu ot co sett nt ha enn ve ETT Sr ey EE ah in SEAiRSNR S D Oa TEn T sa E Moe Tame e S FoRESn a ae Ec iaC lae icenGa e Le e FEE i in RIAEREE i seEitR nC y a TEerawa TE a faEE GoaR RaPRA a Day SY & oh i tt dRTR hr a 03969 l4d% BEST COPY Avs i 1: Appendix AI WATERGOALITYBATA Row oRiCIFTESYT irad | : i 2 Fam ORE oni ada FEEL nGohdiERAER N ni R iBs rhe us STR EES RE SETAE Sa be a A bbO LfdHa lsil n acai BEST COPY AVAII ABLE : T tE si Sbal eceatyrelhinh"es a E TT Te TT 2 2.6 k erreeereeeemeeeseesme Rh eCgnaenmiiE innE co aE onn o cdadE WY RE eeR RE adeDE Se A Ws EE Re Sa CWEe aoin : E CoA E BEE [oe iB i ERR i ee Fu CBT A BESTCOPY AVA) aii: _e hm gm bim g m hem-- cai ain arbi Zoe ein wets wien ees Sot ie Be Leg sa ca CdR fTo aalgE caE PE E reek R i IC e E a aG e RESEE R Sea CT SRD a N SSeEI ASeLe GEL a Sr gl SSaElReTsEREaCOs NG M Su E ET gos i esseiay d]