Document OEzpgb6QXNgMmgxVonw75YGDe

TOXICITY TO AQUATIC PLANTS (E.G.,ALGAE) TEST SUBSTANCE Identity:Perfluorooctylsulfonadtied,ecyldimethylammoniumsalt;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),CAS # 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% of residualperfluorochemicals)A.llvalues reportedrelatetothismixture. The testsample appears tobe a 2-phase dispersion(clearliquiwdith opaque solid)which rapidlyseparatesafteragitationN.o calculations were made toadjustforthe actualconcentrationofthe testsubstance in the testsample. METHOD Method: OECD 201 Test: Staticacute GLP: No Date exposure completed: 1/11/97(studyrepeatedon 1/13/97s;econd studyexposure completed 1/17/97) Species: Selenastrumcapricomutum Source: Originallfyrom The American Type CultureCollectio(nATCC), Strain22662. Analyticalmonitoring: Nominal concentrations.PH, conductivitya,nd temperaturewere monitored. Element basis: Growth rateand number ofcells/mL. Exposure period: 96-hours Startdate: 1/7/97 End date: 1/11/97 Test organisms laboratoryculture:Algae cultureswere growing in OECD-recommended culturemedium atAScl Corporation,Duluth,MN for 4 days beforetestinitiation. Test Conditions: Test temperature range: 25.6- 26.OOC Dilutiownatersource:The algalmediumwas preparetdoOECD recommended concentrationsby spikingdeionizedwaterwith nutriensttocks (includinNga2EDTA). The medium was sterilized via0.22 pm filtratiporniortouse inthe test.The pH ofthe synthetic algalmedium attestinitiatiwoans 7.89. Stock and testsolutionpreparation: Water accommodated fractionsT.est solutionswere preparedforeach testconcentration by mass additionoftestsubstance in500 mL ofalgalmedium. The testsubstance was thoroughlyhomogenized beforeeach mass was determined. The solutionswere vigorouslystirrefdor20-hours (vortex1/2- 1/3solutiondepth).The upper layeroftheaqueous phase was pipettedfrom the vesselsafterthe solutionshad settled for1-hour.The testsolutioninthe highesttwo exposure concentrations2,4 and 40 mg/L were a cloudymilkishcolor,while the 14.4mg/L and 5.2mg/L concentrationswere clear,and the 8.6 mg/L concentratiownas slightlcyloudy.A whiteprecipitatweas alsoobserved on the bottom oftheflasksat allloading concentrationsduringWAF preparation. Exposure vessels: Sterile250 mL borosilicatEerlenmeyer flasks plugged withfoam stopperscontaining100 mL oftestsolution. Agitation:Shaken continuouslyat 100 rpm Number of replicates:Sixforcontrolt,hreeper testsubstance loading Initiaallgalcellloading: 1.0 X 104 cells/mL Number of concentrations: Fiveplusa negativecontrol Water chemistry: pH range (0- 96 hours) 7.89- 9.84 (controelxposure) 8.13- 9.32 (40mg/L exposure) Conductivityrange (0- 96 hours) 131 - 150 pmhos/cm (controelxposure) 134 - 156 pmhos/cm (40mg/L exposure) Test temperature range (0- 96 hours) 25.6 - 26.OOC Light levels:(0 - 96 hours) 730- 750 ft-cfrom continuouscool-whitfeluorescent lighting Remarks: Inconsistentgrowth inhibitioantincreasingexposure concentrationswas seen, particularaltythe 8.6 mg/L exposure concentration.As a resultt,he 48 and 72-hour EL50 based on cellgrowth were lessthan the lowestexposure concentratio(n5.2mg/L),whilethe 48, 72, and 96-hour EL50 based on growth ratewere greaterthan the highest exposure concentratio(n40 mg/L). The NOEL (celdlensity)was also lowerthan the lowestexposure concentration. .RESULTS Nominal concentrations: Bk control,5.2,8.6,14.4,24, 40 mgll. Element value: 24-hour EL5o (celldensity) 15.8(13.2- 18.8)mg/L 24-hour E,L5o(growthrate) 15.9(14.4-17.6m)g/L 48-hour EL5o (celldensity) <5.2mg/L (Clnot calculable) 48-hour E,Lso(growthrate) >40 mg/L (Clnot calculable) 72-hourELso (celldensity) <5.2 mg/L (Clnot calculable) 72-hourErL50(growthrate) >40 mg/L (Clnot calculable) 96-hourEL5o (celdlensity) 20.8(16.4- 26.4)mg/L 96-hour E,L5o(growthrate) >40 mg/L (Clnot calculable) 96-hourNOEL (celdlensity):1.9(1.6- 2.5)mg/L* 96-hourNOEC (growthrate):18.9(14.6- 22.1)mg/L* *NOEL values are approximationsusingthe [CIOas the endpoint. Allelement valuesbased on nominal concentrations. Statisticamlethods: Celldensitiesg,rowth ratesand percentinhibition valueswere calculatedusingan Excelspreadsheet.The Trimmed Spearman-Karber method and the InhibitioCnoncentration(lCp)approach (Version2.01,USEPA-ERL, Duluth,MN) were used forEL and NOEL determinations. Control response: satisfactory Biologicalobservations after96-hours: Nominal Loading Test Concentration, mg/L Mean Number of Cellsper mL Percent Percent Inhibitiovnia Inhibitiovnia Density Growth Rate Blank Control 6,930,000 - - 5.2 4,330,000 -38 -7 8.6 5,720,000 -17 -3 14.4 4,220,000 -39 -8 1 24 1 3,020,000 1 -56 1 -13 40 2,500,000 -64 -16 Observations: Algalcellcounts ineach testvesselwere determined by means ofdirectmicroscopiccounts withan Improved Newbauer hemacytometer. CONCLUSIONS The testsample96-hourEL50intervfaolrSelenastruCmaPricomutuwmas determineudsingtwocalculatimoenthods.By celdlensityi,twas 20.8 mg/L witha 95% confidenceintervaolf 16.4- 26.4 mg/L, and by growth rate>40 mg/L (Clnot calculable)T.he 96-hourNOEL was determinedto be 1.9mg/L usingcelldensityand 18.9mg/L using growth rate. Submifter: 3M Company, EnvironmentalLaboratory,P.O. Box 33331, St. Paul,Minnesota,55133 DATA QUALITY Reliability:Klimischranking3. The study lacksanalyticamleasurement oftestsubstance concentrationsinthetestsolutionsand sample purityis not sufficientclhyaractehzed.Accurate EL5o'scould notbe calculatedat each observationperioddue tothe inconsistendtose-response relationshiapnd low inhibitionbserved atthe exposure concentration loadings.Additionalldya,ta isfora mixtureand toxicitcyannot be positivelayttributetdo didecyldimethylammoniumPerfluorooctylsulfonate saltalone. Report quality:Two definitisvteudieswere conducted on thistest substance atthe same laboratoryi,nthesame time period.A careful examinationofthe raw datarevealedseveralerrorsduringreport preparation.These are: 1 . The "studycompleted"dates printedon the reportcovers (January29, 1997 and February21, 1997) are switchedwith each other.The sectionsunder "1.0General Informationf"or "DefinitivTest Dates"are correctforeach report,however. 2. The discussionunder section"3.0Test Methods" for"Test loadings"are switched. However, thetestconcentrationslisted inthissectionare correctineach report. 3. The algicidal/algistdaettiecrminationtestwas onlyconducted afterthesecond study. The discussionunder "4.0Results"in the reportwiththe cover readingJanuary 29, 1997 should not have been included.No evidencewas seen inthe raw data that the algicidal/algistdaettiecrminationwas conducted duringthe firssttudy.The raw data from the second studyrevealedthat the testsubstance was, infact,algicidal. REFERENCES This study was conducted atASci Corporation,EnvironmentalTesting DivisionD,uluth,MN atthe requestofthe 3M Company. OTHER Last changed: 5/24/00 AScl Corporation/AS[c-Duluth EnvironmcntaTlcstingDivision AScl StudyID# 5olo-o68 STUDY TITLE GROWTH INHIBITION OF TO GREEN ALGA (Sel-enastnoncapricomutum) DATASTANDARD OECD GUIDELINE 201 STUDY COMPLETED February 21, 1997 TESTING FACILITY ASCI Corporation/AScI-Duluth Environmental TestingDivision 4444 AirparkBoulevard Duluth,MN 55811 Tel.No. (218)722-4040 Fax No. (218)722-2592 STUDY IDENTIFICATION NUMBERS ASCI Study ID# 5010-068 3M Company Lab Request# P3025 Sponsor: 3M Company Sponsor Study ID# P3025 ASci Corporation/AScl-Duluth EnvironmentTaelstinDgivision ASclStudyID# 5010-068 1-0 GENIERAL LNFORNIATION StudyTitle DataStandard Sponsor Sponsor's Representative Sponsor's Lab Request# TestingFacility PrincipalInvestigator ProjectDirector TestingFacilityDirector DefinitiveTest Dates Data Validationand Report Review GrowthInhibitiofn p3c-@@ toGreenAlga (Selenastrucmapricornutum). OECD Guideline201. 3M Environmental Technology and Safety Services,Building2-3E-09, 935 Bush Avenue, St.Paul,MN 55133; Tel No. (612)778-7452 Susan Beach, 3M Environmental Technology and SafetyServices,Building2-3E-09, 935 Bush Avenue, St.Paul,MN 55133; Tel No. (612)778-7452. P3025. ASCI Corporation/AScl-DuluthEnvironmental TestingDivision,4444 Airpark Boulevard, Duluth,MN 55811; Tel.No. (218)722-4040. Todd McGuire Joe Amato Donald Mount January 7-11, 1997. Alan Mozol epQ.rtSiLYnature Retentionof Raw Data and FinalReport Locationof Raw Data and FinalReport Two yearsfrom thedate study iscompleted. ASCI Corporation/AScI-DuluthEnvironmental TestingDivision,4444 Airpark Boulevard, Duluth,MN 55811; Tel. No. (218)722-4040. S ponsor: 3M Company Sponsor Study [D# P3(r-S 2 2.0 OBJECTIVE(S) ASci Corporation/AScDlu-luth EnvironmentaTlestingDivision ASci StudyID# 5010-068 To determinethe4-d Medianeffecltoading(ELSO)and the4-dno observableeffecltoading (NOEL) of the testsubstance, capticomutwn) under statitcestconditions. forgreen alga(Selenastrum This stud was conducted accordingto OECD Guideline201 (OECD 1993). Test solution preparationwas water-accommodated fraction(WAF) per Girling& Whale 1994. TestSubstance 3.0 TEST NMTHODS Properties: P.3t two phase liquid,specific gravityca. 1.2 (water= 1),stable,and insolublein water. Testloadings:0 (algalmedium control)5,.2, 8.6,14,4,24, and 40 mg/L. Test loading range was determined from range-findingtestresultsa,nd resultsof previousdefinitivteestsin which inappropriatceoncentration ranges were used. Each testloading was preparedby mass additionof test substanceto 500-ml aliquotsof algalmedium. Test substancewas thoroughly homogenized beforeeach mass was determined. Each solution was vigorouslystirred(vortex1/2 of flasks'depth)for20 hours then allowed to settlfeor one hour to obtainWAF'S. During WAF preparation, a white precipitatweas observed at allloadings,except the 14.4 mg/L loading. A second flaskloaded at 14.4 mg/L was alsosetup, and this second solutionproduced theprecipitate.Both 14.4 mg/L loadingswere tested,and both produced essentialltyhesame results.The presenceor lack of precipitatiodnuring WAF preparationdid not appear to affectthe testsubstancetoxicity.Test solutionswere obtainedfrom flasksby pipettingoff upper layeraqueous phase. Sponsor: 3M Company Sponsor Study ID# P3025 3 Test Organisms AlgalMedium Exposure Chambers Incubation Observations Endpoint Calculations ASci Corporation/AS[c-Duluth Environmental Testing Division ASci Study [D# 5010-068 Source: ATCC 22662 Inoculum preparation: Algal stockwas subculturedin OECD recommended medium for4 days beforetestinitiation. Use in test:Stock culturewas dilutedand spikedintotestsolutiontso obtainan initiaclelldensityof 1.0 X 104 cellsper ml (1.0X 104 actual concentration). Preparation:Deionizedwater was spikedwith nutrientstocks(including Na2EDTA) toOECD recommended concentr'ationsT.he most recent chemical analysisfordeionizedwater isinRaw Data Package. CharacteristicsA:t testtime,the medium had a pH of 7.89. Sterilizevdia 0.22 filtratiporniortouse inthetest. Type: Sterilize2d50-ml borosilicate-glEarslsenmeyer* flaskssealedwith foam stoppers. Use in test: 100 ml of algalmedium or testsolution/chamber.Six control replicateasnd threereplicatepser testsubstanceloadingwere employed. During the test,kept stoppered,exceptwhen experimentalobservations were made. Duration: 4 days. Daily photoperiod: Continuousat740 ft-candleussingcool-white fluorescenltamps. Test chambers heldon shakertablesetat 100 rpm. emperature: 25.0 -+ 20C. Biological:Daily cellcountsusing directmicroscopicenumerationvia Improved Neubauer hemacytometer. Water chemistry: (1)At testinitiatioand terminationp-H, conductivity, and temperature,(2)daily-temperature.Alldeterminationwsere made accordingto APHA methods (1995). 1-,2-, 3-, and 4-d EL50'S, and 4-d NOEL basedon both cellcountsand averagespecifigcrowth rates(/i). Sponsor: 3M Company Sponsor Study ID# P3025 4 ASci Corporation/AScl-DuluLh Environmental TcstingDivision ASel Study ID# 5010-068 4.0 RESULTS Test Substance Toxicity(Pertinendtataare in Tables 1, 2, and 3) Test Conditions CellCount 1-d EL50: 15.8 mg/L (13.2-18.9). A 1-d EL50: 15.9 mg/L (14.4-17.6). CellCount 2-d EL50: < 5.2 mg/L. A 2-d EL50: > 40 mg/L - CellCount 3-d EL50: < 5.2 mg/L. ii3-d EL50: > 40.mg/L. CellCount 4-d EL50: 20.8 mg/L (16.4-26.4). 4-d EL50: > 40 mg/L. CellCount 4-d NOEL: 1.9 mg/L (1.6-2.5). A 4-d NOEL: 19.9mg/L (14.6-221.). NOEC valuesare approximationsusing the IC,o as the endpoint. Effectof testsubstancewas determinedto be algicidalbased on the resultsof a post-test determinationwhere cellspreviouslyexposed to a 400 mg/L loadingfailedto increasein number after9 days. These definitivteest resultswere not appliedto determinethe EL50's as thefinalcellcountsforallloadings resultedin greaterthan 50% inhibition. Temperature (*C): 25.6-26.0. Light intensitfyt-cdls:730-750. pH: 7.84-10.26. 1Specificonductivit(ypmhos@@. 131-160. Sponsor: 3M Company Sponsor Study ID# P3025 5 5.0TEST QA/QC ASci Corporation/ASci-Duluth EnvironmentaTlestingDivision ASclStudy [D# 5010-069 QA/QC Criteria Algalcountsincontrolmust increaseby at leasta factorof 16 duringthefirs3t-d of the exposure. Testdurationmust be 4-d. Data During thefirs3t-d of thetestcontrolalgal countsincreasedby a factorof approximately 388. The testdurationwas 4-d. 6.0 REFERENCES American PublicHealthAssociation(APHA). 1995.Standard.Methods fortheExaminationof Water and Wastewater.APHA, Washington,D.C. OrganizatiofnorEconomic Cooperationand Development (OECD). 1993. OECD Guidelinesfor Testingof Chemicals.OECD PublicatioInnformationCenter,Washington,D.C. Girling,A.E., G.F. Whale, and D.M.M. Adema. 1994. A GuidelineSupplementfor DeterminingtheAquaticToxicityof PoorlyWater-SolubleComplex MixturesusingWaterAccommodated Fractions.Chemosphere, Vol. 29, No. 12,pp. 2645-2649. Hamilton,M.A., R.C. Russo,and R.V. Thurston,1977. Trimmed Spearman-KarberMethod forEstimatingMedian LethalConcentrationisnToxicityBioassays.Environ.Sci.Technol. 11(7):714-719;Correctio1n2 (4):417 (1978). US EPA. 1993. A LinearInterpolatiMoenthod forSublethalToxicity:The Inhibition Concentratio(nlCp)Approach (Version2.0). EnvironmentalResearchLaboratory,Duluth, Minnesota. Sponsor-. 3M Company Sponsor Study ID# P3025 6 ASci Corporation/AScl-Duluth Environmental Testing Division ASci Study [D# 5010-068 TABLE 1. ELSO'S, 95% ConfidenceIntervalsa,nd 4-d NOEL'S from AlgalGrowth Inhibition Test with Endpoint. Cell Count Time 1-d ELSO 2-d F-Lso 3-d F-Lso 4-d EL50 4-d NOEL Data" (MGIL) 15.8 < 5.2 < 5.2 20.8 1.9 95 % Confidence Interval(mg/L) 13.2-18.8 Not Calculable Not Calculable 16.4-26.4 t 1.6-2.5 Average Specific Growth Rate (.u) aW T' (mg IL 1-d EL50 2-d EL50 3-d ELio 4-d EL50 15.9 >40 > 40 >40 .4-d NOEL 18.9 :95@%.C ozifidence. Interval(mg(L).:: 14.4-17.6 Not Calculable Not Calculable Not Calculable 14.6-22.1 a ICIO Values Determined by ICp Program Used to Approximate NOEC's Sponsor-. 3M Company Sponsor Study ID# P3025 7 AScl Corporation/AS[c-Duluth EnvironmentalTestingDivision AScl Study(D# 5olo-o6a TABLE 2. Cellsper ml and PercentageDifferencefrom Nlean ControlValue forSelenastrum capticornutumExposed to T6@t Loading (mg/L) Control 5.2 8.6 14.4 24 Replicate A B c D E F Mean A B c Mean A B c Mean A B c Mean A B c Mean A B c Mean 1d 6 5 3 5 4 4 4.5(-) I 1 4 2.0(-56) 2 3 2 2.3 (-48) 4 2 3 3.0(-33) 1 2 1 l@.3 (-70) 0 2 1 1.0 (-78) C e JIS/Mi (104) 2d 3d 52 55 53 51 46 54 51.8(-) is 19 22 18.7(-64) 25 23 25 24.3 (-53) 17 17 15 16.3(-68) 9 11 11 10.3(-80) 8 10 10 9.3 (-82) 396 378 420 354 372 409 388 105 145 95 115(-70) 245 240 245 243 (-37) 92 ill 94 99(-74) 55 52 53 53 (-86) 46 59 66 57(-85) 4d 649 638 759 682 792 638 693 396 440 462 433(-38) 506 572 638 572(-17) 363 418 484 422(-39) '-776 300 330 302(-56) 204 240 306 250(-64) ParentheticValues % Differencefrom Control Me-an Value 0-d Mean Algal Count = 1.0 X 104 Sponsor: 3M Company Sponsor Study ID# P3025 TABLE3. ASci Corporation/AScl-Duluth EnvironmentalTestingDivision AScl Study ID# 5010-068 Average SpecificGrowth Rates (ita)nd PercentageDifferencefrom Mean Control Value for SelenastrumcapticornutumExposed to T LL di@ Control Replicate Mean 1d 1.504 Average Specifio-GrowthRate (,u)s d 3d 1.974 1.987 4d 1.635 5.2 Mean 0.693 (-54) 1.464 (-26) 1.582(-20) 1.518(-7) 8.6 Mean 0.833 (-45) 1.595(-19) 1.831(-g) 1.587(-3) 14.4 Mean 1.099(-27) 1.396 (-29) 1.532(-23) 1.511(-g) 24 Mean 0.262(-93) 1.166 (-41) 1.325 (-33) 1.428 (-13) 40 Mean 0(-100) 1.115 (-44) 1.348(-32) 1.380 (-16) a ju= In CcUs Per ml !6- in CcUs/ml tn - to b ParentheticValues = % Differencefrom ControlMean Value 0-d Mean Algal Count = 1.0 X 104 Sponsor- 3M Company Sponsor Study ID# P3025