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ACUTE TOXICITY TO AQUATIC INVERTEBRATES (E.G.,DAPHNIA) TEST SUBSTANCE Identity:Perfluorooctylsulfonadtied,ecyldimethylammonium salt;may alsobe referredtoas Fluoroalkyalmmonium derivative[.1Decaminium, N-decyl-N,N-dimethyl-s,altwith 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8-heptadecafl-uoro-1 octanesulfoniaccid (1:1)C,AS # 251099-16-8] Remarks: The 3M productionlotnumber was Lot 1. The testsample is L-14394 referredto by the testlaboratoryas P3025. The sample was labeledF-11615, Lot 1. The testsample isa mixtureofthe testsubstance inwater (approximately30-40% testsubstance,60-70% water,and 0-5% ofresidualperfluorochemicals)A.llvalues reportedrelateto thismixture. The testsample appears tobe a 2-phase dispersion(clearliquiwdith opaque solid)which rapidlyseparatesafteragitation.No calculations were made to adjustforthe actualconcentrationofthe testsubstance in the testsample. METHOD Method: OECD 202 Test type: Staticacute GLP: No Year Completed: 1996 Species: Daphnia magna Analyticalmonitoring:DO, pH, temperatureand conductivitwyere monitored daily. Statisticamlethods: EL5o values calculatedusing Trimmed SpearmanKarber method. NOEL value calculatedusing Steel'sMany-One Rank test. Test daphnid source: Obtained from culturesmaintained by AScl Corporation,Duluth,MN. Test daphnid age at study initiation<: 24-hours Test conditions Dilutionwater: DechlorinatedCityof Duluth,MN tap water. Water was aeratedfor24-hourspriorto use inthe test. Dilutionwater chemistry: hardness: 44 mg/L as CaC03 pH: 8.04 LightingC:ool-whitfeluorescebnutlbs.Photopedodof16-hours light8,-hoursdark. No transitiopneriodnoted. Stock and testsolutionspreparation: Water-accommodated fractionsT.est solutionswere prepared individualfloyreach concentrationby mass additionofvigorouslyshaken testsubstance in1 L ofdilutiownater. The solutionswere vigorouslystirrefdor23hours (vortex1/2to 1/3solutiondepth).The aqueous phase was siphoned from thevesselatmid-depth aftersettlinfgor1-hour. Exposure vessels:250 mL borosilicatgelassbeakers containing 200 mL oftestsolution.The solutionswere kept coveredduring the test. Number of replicates:Four Number of daphnids per replicate:Five Number of concentrations:fiveplusa negativecontrol Water chemistry during the study: Dissolved oxygen range (0- 48 hours): 8.6- 9.1mg/L (controelxposure) 8.0- 8.8mg/L (50mgll exposure) pH range (0- 48 hours) 8.04-8.11 (controelxposure) 7.92- 8.00 (50mg/L exposure) Test temperature range (0- 48 hours) 20.9 - 21.OOC Conductivity range (0- 48 hours): 142 - 155 pmhos/cm (controelxposure) 120 - 124 pmhos/cm (50 mg/L exposure) Element basis:mortalitaynd immobilization RESULTS Nominal mg/L Element loading concentrations: Bk control,3.13,6.25,12.5,25,50 value: 24-hour EL50 = 27.0 (18.7-39.0)mg/L 48-hour EL50 = 11.3(9.6-13.2m)g/L 48-hour NOEL = 6.25 mg/L Allelement valuesbased on nominal concentrations. StatisticaElvaluation:The EL50 valuesand 95% confidenceintervals were calculatedby the Trimmed Spearman-Karber method. The NOEL was calculatedusingSteel'sMany-One Rank testusingthe TOXSTAT statisticsaolftwareVersion 3.2,UniversitoyfWyoming. BiologicaolbservationsD:aphnidsinthenegativceontroaln,d the3.13 and 6.25 mg/L treatmentsappeared healthyand normal throughoutthe testwithno mortalityi,mmobilitoyr overtclinicaslignsof toxicityT.he effectsnoted inthisstudywere mortalityn;o immobilizatiownas noted at any testconcentration.The number ofsurvivingorganisms were determined visuallaynd recordedinitialalnyd after24 and 48 hours. Cumulative percent m rtality: Nominal Loading Test Conc.,mg/L 24-hours 48-hours Neg. Control 0 0 3.13 0 0 6.25 0 0 12.5 5 70 25 50 95 Control response: satisfactory Remarks: Values reportedare forthe testsample. No calculationwsere made toadjustfortheconcentrationofthe testsubstance inthe test sample. CONCLUSIONS The testsubstance 48-hour EL50 forDaphnia magna was determined to be 11.3 mg/L witha 95% confidenceintervaolf9.6-13.2mg/L. The 48hour no observed effectlevel(NOEL) was 6.25 mg/L. Submitter:3M Company, EnvironmentalLaboratory,P.O. Box 33331, St. Paul,Minnesota,55133 DATA QUALITY ReliabilityK:limischranking2. The studylacksanalyticamleasurement oftestsubstanceconcentrationsinthe testsolutionsand sample purityis not sufficientclhyaracterizedA.dditionallyd,ata isfora mixtureand toxicitcyannot be positivelayttributetdodidecyldimethylammonium Perfluorooctylsulfonastaeltalone. REFERENCES This studywas conducted atAScl Corporation,EnvironmentalTesting DivisionD,uluth,MN, attherequestofthe 3M Company. OTHER Last changed: 5/24/00 ASciCorpomlion]AScl-Duluth EnvimntnertTteAsLtIinDgivision ASciStudyID# 5010-069 STUI)Y TITLE ACUTE TOXICITY OF DEVELOPMENTAL TO DAPHNL4 MAGN.4 MATERIAL DATASTANDARD OECD GUIDELINF- 202 STUDY COh4ELETED January 10, 1997 TESTING FACILITY ASCI Corporation/ASel-Duluth EnvironmentalTestingDivision 4444 AirparkBoulevard Duluth,MN 55811 Tel.No. (218)722-4040 Fax No. (218)722-2592 STUDY IDENTIFICATION NUMBERS ASel Study ID# 5010-068 3M Company Lab Request# P3025 Sponsor: 3M Company Sponsor StudyID# P3025 ASci Co rpomtion/AScl-Duluth Environmental TestingDivision ASci Study ID# 5olo-o68 1.0 GENERAL MORMATION StudyTitle Data Standard Sponsor Sponsor'sRepresentative Sponsor'sLab Request# TestingFacility PrincipalInvestigator ProjectDirector TestingFacilitDyirector DefinitiveTest Dates Data Validationand ReportReview AcuteToxicitoyf Developmental MaterialtoDaphnia magna. OECD Guideline202. 3M EnvironmentalTechnology and Safety Services,Building2-3E-09,935 Bush Avenue, St.Paul,MN 55133. Susan A. Beach, 3M Environmental Technology and SafetyServices,Building23E-09,935 Bush Avenue, St.Paul,MN 55133; Tel No. (612)778-7452. P3025. AScl Corporation/AScl-DulutEhnvironmenL-d TestingDivision,4444 AirparkBoulevard, Duluth,MN 55811;Tel.No. (218)722-4040. Joe Dierkes. Joe Amato. Donald Mount. December 11-13,1996. Alan Mozol. Re rtsignature: Retentionof Raw Data and FinalReport Locationof Raw Data and FinalReport Two yearsfrom thedatestudyiscompleted. ASCI Corporation/AScl-DulutEhnvironmental TestingDivision,4444 AirparkBoulevard, Duluth,MN 55811;Tel.No. (218)722-4040.--Jl Sponsor: 3M Company Sponsor Study ID# P3025 2 ASci Corporation/ASIc-Duluth Environmental Testing Division ASci Study ID# 5010-068 2.0OBJECTIVE(S) To determintehe48-hELSO(effectliovaedinga)nd48-hNOEL (noobserveedffecltoadingo)f thetesstubstancLe-,14394DevelopmentMaalteriaflo,rDaphniamagna. Thisstudywas conducteadccordintgotheASCIstudyplanretainbeyd theSponsoarndOECD Guideline203 (OECD 1993). The testsolutionwsere water-accommodatedfraction(sWAF) preparedfollowingGirlingW,hale, & Adema 1994. 3.0 TEST METHODS TestSubstance Test Organisms DilutionWater Propenies:f@c)@@ two-phaseliquidbeforeagitatedl,ightcolored,nearly odorlessslurryafteragitations,pecifigcravityca.1.2g/cc,and insoluble in water. Testloadings:0 (dilutiownater control)3,.13 mg/L, 6.25 mg/L, 12.5 mg/L, 25 mg/L, and 50 mg/L. Testloadingsolutionwsere prepared individuallfyoreach concentration.The solutionwsere preparedby mass additionof vigorouslyshaken testsubstancein 1 L of dilutiownater. The solutionwsere vigorouslystirre(dvortex1/3to 1/2solutiodnepth)for 23 h. The aqueous phase was siphonedfrom thevesselatmid depth. Source: .Healthy,lessthan 24-h old Daphnia magna neonatesobtained from AScl stockculturesmaintainedinwaterof testdilutiownaterquality. Use intest:Gravidadultsisolatedin dilutiownater24 h beforethetest initiationU.sed 5 organisms/replicaatned 4 replicates/tceosntcentration. Handled witha smooth bore pipetwithan insidediameterof 6 mm. Did not feedduringthetest. Source: DechlorinateCdityof Duluth,MN, tapwater. The most recent chemicalanalysisisinRaw Data Package. CharacteristicsA:t testtime,the water had a hardnessof 44 mg/L (as CaC03) and a pH of 8.04. Aeratedfor24 h priortousinginthetes Sponsor: 3M Company SponsorStudy ID# P3025 3 ASel Corporation/AS[c-Duluth Environmental Testing Division ASci Study [D# 5010-o6a Exposure Chambers Incubation Observations Endpoint Calculations 7@pe:250-mlborosilicgaltaesbseakers. Use intest: 200 ml of dilutionwater or testsolution/chamber.During the test,kept covered,exceptwhen experimentalobservationswere made. Duration:48 h. Dailyphotoperiod: 16 h lightand 8 h dark,maintainedusingcool-white fluorescenbtulbs. Temperature: 21*C. Biological:Daily- gentlyagitatedtheexposurechambers and thencounted immobilizedand dead testorganisms(immobilizedorganismswere those thatdid not swim within15 secondsafterexposurechambers were gently agitated).Did not remove any daphnidsfrom theexposurechambers at 24 h. Water chemistry:(1)At testinitiatiotno-talhardnessand alkalinity(,2) daily-temperaturep,H, DO, and conductivityA.lldeterminationwsere made accordingtoAPHA methods (1995). ELSO: Trimmed Spearman-Karbermethod. NOEL: Steel'sMany-One Rank Test (TOXSTAT statisticsaolftware, Version3.2,Universitoyf Wyoming). Sponsor: 3M Company Sponsor Study ID# P3025 4 ASciCorpomtion/ASci-Duluth EnvironmentTaclstinDgivision AScl StudyID# 5010-06S 3.0RESULTS TestSubstancTeoxicit(yPertinednattaarein Table 1) Ranges of Water Chemistry Parameters 24-hEL50:27.0mg/L (18.7-39m.g0/L). 48-h EL50: 11.3 mg/L (9.6-13.2mg/L). 48-h NOEL: 6.25 mg/L. Totalhardness (asCaC03 mgll): 41-44. Alkalinity(asCaC03 mgll): 42-45. Temperature (*C): 20.9-21.0*C. DO (mglL): 7.9-9.1. pH: 7.92-8.11. Specificonducti@@ (,gmhoslcm):120-155. 4.0 TEST QA/QC -QA/QC Criteria Test organism dead,immobilizedor trappedin controlexposuremust not be more than 10%. The lowestDO concentratioanttest terminatiomnust be atleast60% of theair saturatiovnalueat testtemperature. Test durationmust be 48 h. Referencesubstancetoxicittyest (thistestis optional). Data 0% dead,immobilizedor trapped. The lowestDO concentratiomneasured was 89% of theairsaturatiovnalueat21.0"C. The testdurationwas 48 h. CurrentcontrolchartforD. magna acute testingisincludedin the raw datapackage. Sponsor: 3M Company Sponsor Study [D# P30,-15 5 ASci CorporationlAScl-Duluth EnvironmentTaelstinDgivision ASciStudyID# 5010-06g 5.0REFERENCES AmericanPublicHealthAssociati(oAnPHA).1995.StandarMdethodsfortheExaminationf Water and Wastewater.APHA, Washington,D.C. Girling,A.E., G.F. Whale, and D.M.M. Adema. 1994.A GuidelineSupplementfor DeterminingtheAquaticToxicityof PoorlyWater-SolubleComplex MixturesUsing WaterAccommodated FractionsC.hemosphere 29(12):2645-2649. Hamilton,M.A., R.C. Russo,and R.V. Thurston.1977.Trimmed Spearman-KarberMethod for EstimatingMedian LethalConcentrationisn ToxicityBioassays.Environ.Sci.Technol. 11(7):714-719C;orrectio1n2(4):417(1978). OrganizationforEconomic Cooperationand Development (OECD). 1993.OECD Guidelinesfor Tes ng 0 Chemicals.OECD PublicatioInnformationCenter,Washington,D.C. Universityof Wyoming. ToxstatVersion3.2.Cheyenne, Wyoming 82001. Sponsor: 3M Company Sponsor Study ID# P30'25 6 ASci Corporation/ASci-Duluth Environmental Testing Division AScl Study ID# 5010-@8 TABLE 1. ACUTE TOXICITY OF P'@c4 DEVELOPMENTAL MAGNA MATERIAL TO DAPHNIA mg/L) 0 3.13 6.25 12.5 25 50 T C=Wative' Number and Percentof AffectedOrganisn3s' 24. h 48 h 0(0) 0(0) 0(0) 1(5) 10(50) 15(75) 0(0) 0(0) 0(0) 14(70) 19(95) 20(100) a Percentagesare in parenthesis. EFFECT AND NO-EF7ECT LOADINGS WITH CORRESPONDING 95% CONFIDENCE INTERVALS on 24-h EL50 48-h F-Lso 48-h NOEL lba't'i 27.0 11.3 6.25-- 95 % Confidence Interval 18.7-39.0 9.6-13.2 Not Applicable Sponsor: 3M Company Sponsor Study ID# P3025 7