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TOXICITY TO AQUATIC PLANTS TEST SUBSTANCE Identity:A mixturecontainingperfluorooctanesulfonatweh,ich may also be referredtoas PFOS, FC-95, or as a component of FC-203CE. (1Octanesulfonicacid)(CAS # 2795-39-3). Remarks: The 3M productionlotwas notnoted.The testsample isFC203CE. Currentinformationindicatesitisa mixtureof 1.84% PFOS, 30% diethyleneglycolbutylether,60.23% water,2.56% Sultonefoamer, 3.1% sodium octylsulfate2,.1% polyoxyethylenemonooctylphenylether,0.12% sodium laurylsulfatea,nd 0.05% tolyftriazole. The followingsummary applies to a mixture with incompletely characterized concentrations of impuddes. Data may not accurately reflectthe toxkity of the ffuorachemical component of the testsample. METHOD: Method: OECD 201 and EPA-600/9-78-018 Type: Acute Static GLP: No Year study performed: 1991 Species: Selenastrumcapricomutum Source: In-houseculture.Originallfyrom the CultureCollectioonfAlgae at the UniversityofTexas atAustin. Element basis: AJgalcellcounts (cells/mi) Exposure period: 96 hours StatisticaMlethods: Performed usingstandardstatistictaelchniques. Stephan, 1983. NOEC was calculatedusingDunneft'stest. Analyticalmonitoring: pH and temperature Test Conditions: Algal NutrientMedium: Nutrientmedium per U.S. EPA, 1978 prepared in deionizedwater. This nutrienmtedium providedallmineralnutrientesssential foralgalgrowth and also served as the diluentforallalgaloperations.The pH ofthissyntheticalgalmedium was -8.0. Stock and testsolutionpreparation: A 1000 rng/Lprimarystocksolution was prepared inalgalmedium. Aliquotswere added directltyo algalmedium intestvesselsto createtestsolutions. Exposure vessels: 250 mL Erienmeyer flaskscontaining100 mL test solutionand capped withinvertedglassbeakers. Agitation: Continuous rotaryshaker,rateofrotationotgiven. Number of replicates:3 Initiaclellloading: 1.0 X 104 cells/mL. Number ofconcentrationfsi:vpelusa blankcontrol Lighting:Continuoulsightiantg50 pEin/sectmu2singcool-whiftleuorescent lamps. Water Chemistry: pH range (0 - 96 houm): 8.2- 9.9 (controlexposure) 8.3- 8.6(1,000rng/Lexposure) Test temperature range (0- 96 houm): 23.0- 23.4*C (incubator) RESULTS Nominal concentrations: Blank control,150, 250,400, 600, and 1000 mg/L Algal Growth Response at96 Hours Nominal Avg. Coll-Count Avg. Specific Exposure (colls/mL) Growth rate % Change Conc. (mg/L) (p) Control 479,000 0.040 - 150 188,000 0.031 22 250 58,000 0.018 55 400 20,000 0.007 82 600 3,000 -0.012 100 1000 1 0 0.000 100 (1)Average growth rateincontrolcompared totestconcentrationgrowth rate. Element values: 96-hour ErC5o= 230 (212 - 248) nig/L 96-hour NOEC = <150 mg/L Element values are based on nominal concentrations. Revemibilityof Growth Inhibition:Not determined. Remarks: Testingwas conducted on a mixtureas describedinthe Test Substance Remarks field.The valuesreportedapplyto thatmixtureand not the fluorochemicalproportionalone. CONCLUSIONS The testsubstance exhibitsa 96-hour EC5o of 230 mg/L witha 95% confidenceintervaolf212 to248 mg/L.The 96-hourNo Observed Effect Concentration(NOEC) was <150 mg/L. Submitter: 3M Company, EnvironmentalLaboratory,P.O. Box 33331, St. Paul,Minnesota,55133 DATA QUALITY ReliabilityK:Jimischranking= 2. Testingmeets the criterifaorquality testing.However, sample puritywas notpropedy characterizeadnd the study lacksanalyticaclonfimiatioonfthe amount offluorochemicaplroportioinnthe solution. REFERENCES Thistestwas conducted by EnviroSystemsDivisionof Resource Analysts, Incorporatedo,f Hampton, NH atthe requestofthe 3M Company, Lab Request number J1884-1,1991. OTHER Last changed: 6/26/00 staticlcateIroxicito-fi-,rc-203CE to.,thjeaga..soialo@ Robert Tinothy i..v.ard_ ............. study let 7@7 sr@.q. 5i4llp*N Resource,_An Latayett ad' smpton",.x The acute toxicity of rC-203CS to the alga, Selanastram capricorautun, is described in this;final report. The test was conducted for 3N Company for 96 bourn during July 8 to July 12, 1991, at the InviroSystess Division of Resource Inalysts, Inc. in Hampton, Now Hampshire. It was conducted by. J*m@me Nagazu, Peter Kowalski, Ellen ":,,StanfordR,obert fteri, and Timothy The test was performed under static conditions with fiv0 concentrations of test substance and a dilution water control at a mean -teaperature of 23.2*C. The dilution vator was synthetic media prepared with sterile doianized water@ Tlasks":we re'incubated on a rotary shaker. 'Light was adjusted to 50 Wtio/sec/0@il with a photoperiod of 24 hours light and 0 hours dark ' Nominal co rations of IPC-203CSwere: 0 mg/L (control), 150 mg/L, 2iO mg/l., 400 mg/Le,.600 mg/L, and 1,000 no/L. Nominal concentrations were used for all-maculations. kigae used in the test was "from an in-house culture that was iiaintainod under test conditions., Exposure of algae to the test substan.-o ,resulted in a 72 hour IC50 of 340 mg/L IPC-203CEand a no observed offect '.:'concentratio(nNOZC) of 150 mg/L. Th4i'9.h6our EC50 and NOZC are 230 mg/L and less than 150 mg/L, respectively. -3 OX U. TIM CONTZNTX SECTION: PACE 1. sumary 2 ii. Table of.Contents 3 Ill. lndsx of-Tablis 4 IV. Introduction 5 V. Nothodi and Materials Vi. Results"- VIZ. References 12 JLMndix iL.:fs,tar,Qwa t@.y. Data -'rImf Taxicity.'T-e.st 4.0 Ill. nnpa OF Tulin Table 1. Chemical characterizationof a representative' sample of dsionized water used to prepare test media for toxicity test PACE 6 Table 2. Growth data from toxicity test 9 Table 3. Percent change from control and average 10 specific growth rate from toxicity test Table 4. Median effective concentrations (Zcsos) from toxicity test Table &.I. Temperature measured during.,toxicitytest it Table A.2. pg measured during toxicitytest IT. INTROMMON This.@ study was sponsored by 3H Coipiii,St.Paul# Minnesota. The objectiveof the Study was to daterain*the acute toxicityof TC-203CE to a freshwater alga. The report contains sections that describe the methods and materials employed in the study, and the results of the investigation.The report also contains an appendix that presents the water qualitydata collectedduringthe test. V. XCMDS AIM RLTEKMS TEST SUBSTANCE: TC-203CI (SaviroSystess Sample Number 2935A) was delivered to EnviroSystess on May 23, 1991. It was containedin a 40 oz. plastic bottle that was labelled with the following information:"JIS84-1. IrC-203CE".., TC-203CZ (an orange-brown liquid) was supplied by 3H company, 935 Bush. &venue, St. Paul, Minnesota. Prior to use the test material was stored in the dark at room temperature. I reservesample (approximately 1.1 grass) will be retainedat InviroSysteasfor a minimum of 10 years. DUAUTION WATER: Water wed for acclimationof test organismsand for all toxicity testing was sterile enriched media (U.S. EPAO 1978) with a pH of approximately 0.0. Results of chemical analysis of a representative sample of ,deionized water used to prepare the media are presented in Table 1. TEST Oitomsx: algae" used*for the toot was from a cultureorig"iit the' Culturt collection of Algae.@atthe Universityof Texas at Austiji".and sarirosyst- ess oi@Nay 14, 1991o The identity.*ofttb6-Liulture was vorifi*d @iU.sing@ an approprlate taxonomic key "-'@l-'The was media :identicalti@, dia''Used"foet,wls, t tiTc.;t:omicitt@.t-e'st,.T.o, liirfoinid@ 1991 is@ based on@,,p-roc*dur4 OZO, (19 at Yatistei.-.'teiperatloifk2e2,:k 21!C with fly and dilution.'!rart.i,,,contI"r*o-@'-@A',1,000 @a"g/I. at @,,,was..prop'liredfiiv',z'dilutviaotna t'",.i,.T-he :stock,7501utlon was@@added aimed,,: tosr;a,ve,ssels@wi concentrati'6iks,":"Ofx@:iii*,-',test - 250 ag/L;,,400-iag/l;,@.,.,600'ag/'La, to. t' fiat :,,i6itoit rt Tole I Chemical,characterizatioonf'*a, representativeS&RPIO 0 t deionizedwaterused to preparetestmediafor toxicitytest Parameter Unit of loportina Neasureaeut Uikit Reasured Value organochlorine ug/L 2 ND pesticides organophosphoras ug/L o.5 ND pesticides Polychlorinated ug/L 0. 5 ND biphonyls Not*: ND not detected at or above the reporting limit. klgae was randoaly and equally distributed aaong three replicates of each treatmentat the rate-of10,000collsfal. The test was performed in 250 &I glass Uleme"r flasks that contained100 al of test solution. The flasks were capped with invartedglass beakers. Test vessels were randoaly arranged on a rotary shaker in an incubator during the 96 hour test (a random numbers table was used to select the location of each vessel). A 24 hour light and 0 hour dark photoperiod was maintainedwith cool-whitefluorescentlights that provideda light intensityof So UZS-IM-2. The number of algal collslalin each test vesselwas determined visually *very 24 hours by means of direct microscopiccountswith a hemacytonoter. The pl (Backmanmodel pal 51 motor) was determinedin each test flask at the beginningand end of the test, and temperature (KSTX mercury thermometer) was measured and recorded daily in the incubator. STITISTICILL NETRODS: Results of the toxicity test wore interpretedby standard statistical techniques(Stephan,1993). The binomialor moving average method was used by the author to calculateSCSO values using nominal concentrationsof test substance. The VOIC was calculatedusing Dumett's test, which includes a parametric one-way analysis of variane (INOVA). The average specific growth rate was calculated as the Ttural log of the number of cells per al at 24, 48, 72, or 96 hours minus the natural log of the numberof cells par al at 0 hours divided by the owsure@. period. The percent change from control is -calculated by subtractingthe sample averag*'specificgrowthrat* froa ..@,th,o.'@,:conatvreorlage 'specific'g@rowth rate, dividing that value by the ayerage.,ap*cific growthrate :i@@tho'controland multiplying that by: 100., _,Thep.ercent change"f'rom cozktrolv,u used to coaputo ZCSO mosc values.".,- ,,Pagt-7" -Ti. IZMTB Biological and water quality data generated-by the acute toxicity test are presented in Table 2 and App*udixA. respectively, and the percent of control and average specific growth rate information is presentedin Table 3. The 24, 48, 72, and 96 hour BC50s for algae ezp"ed to FC-203CZ are presented in Table 4. The 72 hour ZCSO is 340 ag/L. and the 72 hour NOZC is 150 mg/L FC-203CZ. The 96 hour ZC50 is 230 ag/L and the 96 hour NOIC is less than 150 mg/L. 0 -table2. fromto3dcitY ttst loominal concentration rep. 0.0 @tcoutrol) 150 250 400 600 000 1 2 3 mean 1 2 3 mean 1 2 3 *Gan 1 2 3 seen 1 2 3 "an 3. lijllj'lite2r103 Nusber Of calls per 0 24 48 12 (bour) 96 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 to 10 10 10 10 10 10 10 10 12 Is is 2.6 14 14 10 13 20 20 20 20 14 10 16 13 30 22 20 24' 249 120 106 136 121 64 70 66 @46 46 48 47 54 .40 ILI 4;4 38 38' .430 21 I ..,-1@ 144 219 196 196 as 110 70 89 so 95 is 10 25 .40 25 30 40 20 20 21 ---3S D .-. 21 312 450 61S 419 215 195 iss Is$ 50 to AS se 25 10 25 l'O 0 0 'or. Table 3. P*rcsnt cwmgs from control and average specific growth rate from toxicity test Nominal Concentration of Test Substance (mg/L) Average specific growth rate 24hr 4&hr 72br 96hr 0.0 (control) 150 250 400 600 1,000 0.020 0.011 0.029 0.011 0.037 0.033 0.052 0.040 0.032 0.032 0.034 0.026 0.04i 0.040 0.030 0.031 0.027 0.01$ 0.015 0.007 0.014 -0.012 0.014 0.000 Percent change from control 24hr 4&hr 72hr 96hr 0 0 0 0 45 23 27 22 -45 38 34 55 45 38 63 82 -85 35 66 100 -85 50 66 100 L ? Table 4. ftftu effectivecoiesstratims (MC50s)fron'tmicitytest EWsurs. period zcso 9S percent confidence limits Calculation method 24 hours 48 hours 72 hours 96 hours 11000 mg/L ISO mg/L 340 mg/L 230 mg/L- 247 438 mg/L 212 - 248 mg/L Moving Average Probit Pr ....... ..y as ii;, o, -.@IlL 090. 19$41. cuidelinesfor Testing of Chemicals. Section 2: Effects On Biotic ftstmx. xethod 201, iliga,Growth inhibitionTest. kdopted JunG 4, 1984. Stephen. C.E. 1983. Computer program for calculation of LC50 values. Personal communication. U.S. Vi. 1979. The solonastrm capricorautum Printz klgal Assay Bottle Test. EPI-600/9-18-018.'July, 1978. aii JAN L AROWM MM kippendix A. UTZI OUALITY DATK nOR TOXICITY TIST pass, 11- Table 1.1. Tesperaturesou,-r*dduringtoxicitytest Day of IW sure 0 1 2 3 4 ToW rature of lacubator,OC 23.2 23.0 23.0 23.4 23.2 17,Zillt, .Tabl* A.2. pR measureddaring toxacity test Noainal concentration (ag/L) 0 (control) iso 250 400 600 1,000- Replicate 1 2 3 1 2 3 1 2 3 1 2 3 1 2 3 2,-, 3,- -VIA, initial 8.2 8.2 2.2 9.4 8.4 8.4 8.4 8.4 8.4 8.4 8.4 3.4 8.4 8.4 8.4 8.3 8.3 8.3 final 9.9 9.3 9.9 9.7 3.7 8.7 9.7 3.7 9.7 8.6 9.6 8.6 2.6 8.6 4.6 8.6 8.6 8.6