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Ar226 _ 0886 TOXICITY TO AQUATIC PLANTS TEST SUBSTANCE Identity: Mixture containing perfluorooctanesulfonate which may also be referred to as PFOS, FC-95, or as a component of FC-3003. (1Octanesulfonic acid) (CAS # 2795-39-3). Remarks: The 3M production lot number was not given. The test sample is FC-3003, identified by the laboratory as R1331. Cument information indicates it is a mixture of 1% PFOS, 15% diethylene glycol butyl ether, 79.39% water, 2% Sultone foamer, 1.5% sodium octyl sulfate, 1% sodium decyl sulfate, 0.06% sodium lauryl sulfate, and 0.05% benzotriazole. The following summary applies to a mixture with incompletely characterized concentrations of impurities. Data may not accurately reflect toxicityofthe fluorochemical component of the test sample. METHOD: Method: OECD Guideline 201 Type: Acute Static GLP: No Year study performed: 1997 Species: Selenastrum capricomutum Source: Originally obtained from ATCC, Rockville, MD, maintained in culture medium at AScl-DETD, Duluth, MN Element basis: Algal cell counts (cells/ml), specific growth rates. Exposure period: 96 hours Statistical Methods: Statistical analyses were performed using the Trimmed `Spearman-Karber method. Analytical monitoring: pH, conductivity, and temperature Test Conditions: Algal NutrientMedium: Nutrient medium per OECD. This nutrient medium provided all mineral nutrients essential for algal growth and also served as the diluent for all algal operations. The pHofthis synthetic algal medium was 8.0. Stock and test solution preparation: A 2000 mg/L primary stock solution was prepared by dissolving 700 mg in 350 mL algal medium, followed by mechanical stirring. Each test concentration was made by volume addition of the stock to algal medium. --= Exposure vessels: 250 mi Erlenmeyer flasks containing 100 mi test solution, sealed with foam stoppers. Agitation: 100 rpm. Numbeorf replicates: 3 for exposure concentrations, 6 for blank control. Initial cell loading: 1.0 x 10* cells/mL (1.3 x 10* cells/mL actual). 002721 Number of concentrations: five plus a blank control Lighting: Continuous lighting at 740 ft-candles using cool-white fluorescent lamps. `Water Chemistry: pH range (0-96 hours): 8.00 ~ 9.90 (control exposure) 4.72 - 7.90 (1000 mg/L exposure) Conductivity range (0-96 hours): 151 -- 186 pmhos/cm (control exposure) 159 -- 170 pmhos/cm (1000 mg/L exposure) Test temperature range (0-96 hours): 24.8 -25.8 C (incubator) RESULTS Nominal concentrations: Blank control, 130, 216, 360, 600, and 1000 mg/L. Algal Growth ECs, Values Exposure `Average Specific (Contact) Cell-Count Growth Rate Hours mg/L (95% C.1.) mg/L (95% C.1.) |Z | 233 (1--53492) 160 (1--51700) [a8| <130 (Not Calculable) |(225014-308)| [96 C<a<113l300c(N(uoNottlCaalcublabllee) )| 644 (528-834 Element values: 96-hour ECso = <130 mg/L (cell count) 96-hour E.Cso = 664 (528 -- 834) mg/L (growth rate) 96-hour NOEC = 23 mg/L (cell count) 96-hour NOEC = 96 mg/L. (growth rate) Element values are based on nominal concentrations. Reversibility of Growth Inhibition: Aliquotsofthe 1000 mg/L test solution were diluted with algal medium and cultured for an additional period. Based on growth observed in the recovery phase, the effect on algal growth was found to be algistatic. Remarks: Testing was conducted on the mixture as described in the Test Substance Remarks field. The values reported apply to that mixture and not the fluorochemical proportion alone. CT CONCLUSIONS 002722 FC-3003 exhibits Eso growth rate a 96-hour ECso cell valueof664 (528 -- c8o3u4n)tmvga/lLu.eof <130 mg/L. and a 96-hour Tcohuent96a-nhdou9r6Nmog/OL.bfsoerrvgreodwtEfhfercatteC.onTcheinsttreasttisounb(sNtOanEcCe)wiass23demtge/rLmifnoerdcetllo be algistatic. Submitter: 3M Paul, Minnesota, Company, 55133 Environmental Laboratory, P.O. Box 33331, St. DATA QUALITY tReesltiianbgi.litHyo:weKvieirm,isscahmrpalnekipnugri=ty2.waTsesntoitngprmoepeertlsytchhearcarictteerriiafzoerd qaunadlittyhe study lacks analytical solution. confirmationof the amountof fluorochemical proportion in the REFERENCES "DTihviissitoens,t wDualsutcho,nMduNctaetdthbeyrAeSqculesCtoorfpotrhaeti3oMn,CDoumlpuathnyE,nvLiarbonRmeeqntuaelstTensutmibnger R1331, 1997. OTHER Last changed: 6/26/00 002723 EAnSvcitrCoonrmpeonrtaatliToens/AiSncgl-DiDvuilsuiotnh . "Asal Study 104 5010-068 STUDYTITLE GROWTH INHIBITION OF R1331 FOR GREEN ALGA (Selenastrum capricornutum) DATASTANDARD ` OECD GUIDELINE 201 STUDYCOMPLETED April 3, 1997 TESTINGFACILITY AScICorporation/AScI-Duluth Environmental Testing Division 4444 Airpark Boulevard Duluth, MN 55811 TFealx. NNoo.. ((221188)) 772222.-24509420 3 STUDYIDENTIFICATIONNUMBERS AScl Study IDA 5010-068 3M Company Lab Request R1331 SSppoonsoorr:Su3dMy CIoDmFpRanIyI 1 002724 EAnSveilroSCnmolernptoSarlaniyToens1t/0iA1nSgeSloDDliu0vl.iu0st6oh8n 1.0 GENERAL INFORMATION { study Title santas Sponsor `Sponsor's Representative Sponsor's Lab Request `Testing Facility Principal Investigator Director `Testing Facility Director [| oct tonbuee |{ Data Validation and Report Review . RetentionofRaw Data and Final Report | Location of Raw Data and Final Report ep ume, l Growth Inhibitionof R1331 for Green Alga | (Selenastrum capricornutum). 3M Environmental Technology andSafety Services, Building 2-3E-09, 935 Bush Avenue, | St. Paul, MN 55133; Tel No. (612) 778-7452. Susan Beach, 3M Environmental Technology | BaunsdhSaAfveetnyuSee,rvSitc.esP,auBlu,ilMdiNng525-133E3-;09, 935 Il on. Tel No. (612) 778-7452. v [rodameque| AScl Corporation/AScl-Duluth Environmental | Testing Division, 4444 Airpark Boulevard, { Duluth, MN 55811; Tel. No. (218) 722-4040. rm 0.57 Donald Mount Alan Mozol I| byZe Report Signature; | Two years from the date study is completed. | `ATSecstlinCgorDipvoirsaitoino,n/4A4S4c4l-ADiurlpuatrhk EBnovuilreovnamredn,t~al~ || Duluth, MN 55811; Tel. No. 218) 722-4040. | SSphoomoerr: Su3dMyCIoFmpaRn1y31 2 4 002725 EAnSierlonCmopeonrtuaiToens/tAiSnegLD-iDvualiaohn 'AScl Study 104 5010-068 2.0 OBJECTIVE(S) `To determine the 4-d median cuonndceerntsrtaattiicotnes(tNcOoEndCi)tioonfst.he teefsftecstubcsotnacnecnet,raRti1o3n1(,ECfyoyr)graenedn tahleg4a-(dSenloenoabssterruvmabclaeperfifceocrtnutum) This study was conducted according to OECD Guideline 201 (OECD 1993). 3.0 TEST METHODS `Test Substance l `Test Organisms Algal Medium f Exposure Chambers | Properties: (water=1), sRt1ab3l3e1, ,ancdleacromapmlbeetre-lcyolsoorleubdlleiiqnuiwd,atespre.cific gravity=1.02 | { T1e0s0t0comngc/eLn.traAtiosntso:ck 0 (algal solution medium control), was prepared by 130, mass 216, 360, 600, and addition to algal |{ medium made by followed by mechanical volume addition of the stirring. stock to alEgaalchmetedsitucmo.ncentration was | Source: ATCC 22662 Inoculum preparation: recommended medium Algal stock was subcultured in for 4 days before test initiation. OECD "|l Use in rest: Stock culture was diluted and spiked into test solutions to | coobntcaeinntraantiionnit)i.al cell density of 1.0 X 104 cells per ml (1.3 X 10% actual i | Preparation: Na,EDTA) to DOeEioCnDizerdecwoamtemrenwdaesdscpoinkceedntwriatthionnust.rieTnht estmoocskst (including; recent | | chemical analysis for deionized water is in Raw Data Package. 0C.ha2r2acytiemrifsitlitcrsa:tionAtprtieosrt time, to use the medium in the test. had a pH of 8.00. Sterilized via Type: sealed Sterilized with foam via autoclave stoppers. 250-ml borosilicate-glass Erlenmeyer flasks || rUespeliicnatteessta:nd1t0h0remelroepflailcgaatlesmpeedrituemst osrubtsetstanscoeluctoinocne/ncthraamtbieorn.weSriex control | employed. During the test, observations were made. kept stoppered, except when experimental | SSppoonnssoorr:St3udMy CIoDmIpRanyI 3 : 002726 Incubation l Observations | C`aElncduploaitnitons EAnSveilrCoonrmpeonraaliToena/iAnSgelDDivuilsuiotnh 'ASel Sudy DF 5010.068 Duration: 4 days. Daily photoperiod: fluorescent lamps. Continuous at Test chambers 740 held ft-candles using on shaker table cool-white set at 100 rpm. |I Temperaure: 25.0 + 2C. BIimoplroogviceadl:NeuDabialuyercelhlemcaocuynttosmuesticnrg.direct microscopic enumeration via Water chemistry: and temperature, (2()1)daAitlyt-estteimnpietriaattiuorne.and~Altlerdmeitneartmiionna-tipoHn,s wcoenrdeucmtiavdiety, |{ according to APHA methods (1995). | a1-v,er2a-g,e3-s,peacinfdic4-gdroEwCtyh'sra,teasn(d4)4.-dStNaOtiEstCicablaasneadlyosnesboptehrfcoelrlmecdouunstisngand;, `Trimmed Spearman-Karber Method (ECsy's) and ICp analysis. | 4.0 RESULTS Test Substance Toxicity (Pertinent data a=re in Tables 1, 2, and 3) | Ce--ll C--ount I-d ECs:--233 mg/L (159-- -342). - | i 1d ECyg; 160 mg/L (150-170). { Cell Count 2-d ECgy <130 mg/L. | 1 2-d ECs5; 251 mg/L (204-309). { Cel3ldCoEuCsnyti3-Sd7E2Cymyg:/L<(143340-7m5g6/).L. i 1 Cell Count 4-d ECgy: <130 mg/L. lf 4d ECsy: 664 mg/L (528-834). (Ce2l2l24C)o.unt 4-d NOEC (ICI0): 23 mg/L 1 4d NOEC (IC10): 96 mg/L (88-105). { Effect was determined to be algistatic based on production of cells exposed during the i cdeofnicneinttirvaetitoenst. to the 1000 mg/L test substance. `SSppoonnssoorr:St3udMy CIoDmHpRanIyI 4 I 002727 } Test Conditions EAnSveirlC"oAnoSmreeplnotSraualeiToyens1/t0Ai3SnegSloDD1iu0vl.i0us6ot8nh Temperature (C): 24.825.8. 1 Light intensityfi-cdls: 740. . PH: 4.72:9.90. Specific conducivi os/cm): 129-186. | 5.0 TEST QA/QC . [vec come oe +] Algal counts in control must increase by at | During the first 3-d of the test control algal exlpeaosstuaref.actor of 16 during the first 3-d of the | c1o5u7n.ts increased by a factor of approximately Test duration must be 4.4. |a thetest durstioa n was ] 6.0 REFERENCES American Public Health Association (APHA). 1995. Standard Methods for the Examination of ] Water and Wastewater. APHA, Washington, D.C. 3 Organization for Economic Cooperation and Development (OECD). 1993. OECD Guidelines for `Testing of Chemicals. OECD Publication Information Center, `Washington, D.C. : Hamilton, M.A., R.C. Russo, and R.V. Thurston, 1977. Trimmed Spearman-Karber Method for Estimating Median Lethal Concentrations in `Toxicity Bioassays. Environ. Sci. Technol.i.; 11(7): 714-719; Correction 12 (4): 417 (1978). 1 US EPA. 1993. A Linear Interpolation Method for Sublethal Toxicity: The Inhibition : MiCnonnecsenottraa.tion (ICp) Approach (Version 2.0). Environmental Research Laboratory, Duluth, j SSppoonnssoorr: St3udMy CIoDmFpRanIyS 5 . 002728 mcoorsaa TABLE 1. ETCe'stSw,ith95R1%3C3o1nfidence Intervals, and 4-d NOEC'S from Algal Growth Inhibition po Be || m oomm e e ow T |T ewe com] [oi Tm an] wT ----] | pCpRSg [E om T TT w ] ee ewe Te om we iBene es mio : EA Sponsor: 3M Company . 2 002729 TABLE 2. cCealplrsipceormumtlumanEdxpPoesrceednttoagRe13Di3f1ference from Mean Control Value for Selena-strum we | 5AT] l:4 T = 23 200 726 A 2 3- 69 30 | A 2 39 195 I A 3 4 23 7s 1 | 2i ST 1.7 (55) *0-4 MeanAlgalCount =1.3 X 10 10 F2E | 8.7 (:96) :Boesy | i F-- R, . 002730 . EAnvliCoonrmpnoraiToons/iAnSgekDDivuititon, RS Sty 103 s6l0068 TABLE 3. Average Specific Growth Rates (#) and Percentage Difference from Mean Control Value for Selenastrum capricornutum Exposed to R1331 3 TeaCoCmRe EE `Average SpeGrcowithRfatie coF [il =[emer[ee[me [en|{ EERE Lo | ome [osm | wow|omer| *#= lapCele ls rmIinCet llum,i ty * Parenti Vales = bl % Diffs frm Corot Mean Value =04 MeanAlgalCount = 1.X3 10% SSptoonnseorr Su3dMy C1o8mpRanIyI . 0027131