Document X7LGXn7VdgK5OgRL9BXo6Zgqx

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-heptadecafluoro-loctanesulfoniaccid (1:1)C,AS # 251099-16-8] Remarks: The 3M productionlotnumber was Lot 1. The testsample is L-14394 referredtoby thetestlaboratorays P3025. The sample was labeledF-11615, Lot 1. The testsample isa mixtureofthe testsubstance inwater (approximatel3y0-40% testsubstance,60-70% water,and 0-5% ofresidualperfluorochemicals)A.llvaluesreportedrelatetothismixture. The testsample appears tobe a 2-phase dispersion(clearliquiwdith opaque solid)which rapidlyseparatesafteragitationN.o calculations were made toadjustfortheactualconcentratioonfthetestsubstance in the testsample. METHOD Method: OECD 201 Test: Staticacute GLP: No Date exposure completed: 1/17/97(Repeatofearliesrtudycompleted 1/11/97) Species: Selenastrumcapricomutum Source: Originallfyrom The American Type CultureCollectio(nATCC), Strain22662. Element basis: Growth rateand number ofcells/mL. Analyticalmonitoring: Nominal concentrationosnly, pH, conductivity, temperaturewere monitored. Exposure period: 96-hours Startdate: 1/13/97 End date: 1/17/97 Test organisms laboratoryculture:Algae cultureswere growing in OECD-recommended culturemedium atASci Corporation,Duluth,MN for 4 days beforetestinitiation. Test Conditions: Test temperature range: 24.6- 25.6'C Dilutionwater source: The algalmedium was prepared toOECD recommended concentrationsby spikingdeionizedwater with nutrientstocks(includinNga2EDTA). The medium was sterilized via0.22 pm filtratiporniortouse inthetest.The pH ofthesynthetic algalmedium attestinitiatiwoans 7.95. Stock and testsolutionpreparation:Water accommodated fractionsT.est solutionswere prepared foreach testconcentration by mass additionoftestsubstance in500 mL ofalgalmedium. The solutionswere vigorouslystirrefdor21-hours(vortex1/2solution depth).The aqueous phase was pipettedfrom thevesselsatmiddepth afterthe solutionshad settledfor1-hour. Exposure vessels: Sterile250 mL borosilicatEerlenmeyer flasks plugged withfoam stopperscontaining100 mL oftestsolution. Agitation:Shaken continuouslyat 100 rpm Number of replicates:Six forcontrolt,hreeper testsubstance loading Initialgalcellloading:1.2X 104 cells/mL Number of concentrations: Fiveplusa negativecontrol Water chemistry: pH range (0- 96 hours) 7.95- 9.43 (controelxposure) 7.91- 7.78 (400mg/L exposure) Conductivityrange (0- 96 hours) 134 - 150 pmhos/cm (controelxposure) 154 - 166 pmhos/cm (400 mg/L exposure) Test temperature range (0- 96 hours) 24.6 - 25.60C Light levels:(0- 96 hours) 730- 750 ft-cfrom continuouscool-whitfeluorescent lighting Remarks: Inconsistengtrowth inhibitioant increasingexposure concentrationswas seen when the studywas firsrtun using exposure concentrationfsrom 5.2 to40 mg/L. Inthisfirssttudy,the48, 72,and 96hour E,L50(growthrate)could notbe calculatedb,ecause lessthan 50% growth inhibitiownas seen atallexposure concentrations.The study was repeatedathighertestsubstance concentrations.Inthesecond study, complete inhibitiownas seen atevery exposure concentrationloading except the lowesttwo concentrations8,6.4 (growthrate)and 51.8 mg/L (celdlensity,growth rate).As a resultt,he 48,72, and 96-hourEL50 (cell density)could notbe calculatedand the ErL5oateach time periodwas slightlhyigherthan the lowestexposure concentration. RESULTS Nominal concentrations: Bk control,51.8,86.4,144,240, 400 mg/L. Element value: 24-hour EL5o (celldensity)= 55.8 (50.2- 62.1)mg/L 24-hourErL5o(growtrhate) 57.7(54.2-61.m5g)/L 48-hourEL5o(celdlensity)<51.8mg/L (Clnotcalculable) 48-hourErL50(growtrhate=)54.8(50.4-59.m6g)/L 72-hourEL50(celdlensity)<51.8mg/L (Clnotcalculable) 72-hourE,L5o(growtrhate) 61.4(58.2-64.m9g)/L 96-hourEL5o(celdlensity)<51.8mg/L(Clnotcalculable) 96-hourErLro(growtrhate) 64.6(62.5-66.m9g)/L 96-hourNOEL (celdlensity)7:.7(7.5-8.m1g)/L* 96-hourNOEL (growtrhate)2:9.3(27.7-31.m6g)/L* *NOEL valuesareapproximatiounssingtheIC,oas theendpoint Allelementvaluesbasedon nominalconcentrations. Statisticmaelthods: Celldensitiegsr,owthratesand percenitnhibition valueswerecalculatuesdingan ExcelspreadsheetT.he Trimmed Spearman-Karbemrethodand theInhibitCioncentrati(olnCp)approach (Versio2n.01,USEPA-ERL, DuluthM,N) wereusedforEL and NOEL determinations. Controlresponse:satisfactory Biologicaolbservationasfter96-hours: NominalLoading Mean Number Percent Percent Test of Cellsper inhibitiovnia Inhibitiovnia Concentration, mL Density Growth Rate mgil Blank Control 6,130,000 - - 51.8 2,O4OiOOO -67 -18 86.4 20,000 -100 -92 144 10,000 -100 -100 1 240 1 17,000 1 -ioo -94 400 7,000 -100 -100 Observations: Algalcellcounts ineach testvesselwere determined by means ofdirectmicroscopiccounts withan Improved Newbauer hemacytometer. CONCLUSIONS The testsample 96-hour EL5o intervaflorSelenastrum capricomutum was determined using two calculatiomnethods. By celldensity,itwas <51.8 mg/L and by growth rate64.6 (62.5- 66.9)mg/L. The 96-hourNOEL was determined tobe 7.7 mg/L using celldensityand 29.3 mg/L usinggrowth rate. Submifter: 3M Company, Environmental Laboratory,P.O. Box 33331, St. Paul,Minnesota,55133 DATA QUALITY Reliability:Klimischranking3. The study lacksanalyticamleasurement of testsubstance concentrationsinthe testsolutionsand sample purityis notsufficientclhyaracterized.Accurate EL5o'scould notbe calculatedat each observationperioddue tothe almost complete inhibitionfcell growth at allexposure concentrations.Additionalldya,ta isfora mixture and toxicitcyannot be positivelayttributetdo didecyldimethylammonium Perfluorooctylsulfonastaeltalone. Report quality:Two definitivsetudieswere conducted on thistest substance atthe same laboratoryi,nthe same time period.A careful examination ofthe raw data revealedseveralerrorsduringreport preparation.These are: I. The "studycompleted" dates printedon the reportcovers (January29, 1997 and February21, 1997) are switchedwith each other.The sectionsunder "1.0General Informationf"or "DefinitivTeest 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 afterthe second 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 study revealedthat 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 ASelCorporation/A-SDcuiluth EnvironmcntATlcstingDiVi3ion ASci Study ID# 5010-068 STUDY TITLE GROV;TH INHIBITION OF IOjc)K, GREEN ALGA (Selenastnucnapticomutum) DATA STANDARD OECD GUIDELINE 201 STUDY COMPLETED JANUARY 29, 1997 TF,STING FACILITY ASCI Corporation/AScl-D uluth Environmental TestingDivision 4444 AirparkBoulevard Duluth,MN 55911 Tel.No. (218)722-4040 Fa.xNo. (218)722-2592 STUDY IDENTIFICATION NUMBERS AScl Study ID# 5010-068 3M Company Lab Request# P3025 Sponsor: 3M Company Sponsor Study ID# P3025 ASci CorpomtionlAScl-Dut,th EnvironmentaTlestingDivision ASciStudyIDN 501o-o,68 1.0 GENERAL INFORMATION StudyTitle Data Standard Sponsor Sponsor's Representative Sponsor'sLab Request# TestingFacility PrincipalInvestigator ProjectDirector TestingFacilityDirector DefinitivTeest Dates Data Validationand Report Review Growth Inhibitioonf capricomuturn). ,Selenastrwn OECD Guideline201. 3M EnvironmentalTechnology and Safety Services,Building2-3E-09,935 Bush Avenue, St.Paul,MN 55133; Tel No. (612)778-7452. Susan Beach, 3M EnvironmentalTechnology and SafetyServices,Building2-3E-09,935 Bush Avenue, St. Paul,MN 55133; Tel No. (612)778-7452. P3025. AScl Corporation/ASCI-Du tuth Environmental TestingDivision,4444 AirparkBoulevard, Duluth,MN 55811;Tel.No. (218)722-4040. Todd McGuire Joe Amato Donald Mount January 13-17,1997. Alan Mozol ReportSignature: Retentionof Raw Data and FinalReport Locationof Raw Data and FinalReport Two yearsfrom thedatestudyiscompleted. ASCI Corporation/ASclD-uluthEnvironmental TestingDivision, 4444 Airpark Boulevard, Duluth, MN 55811; Tel. No. (218)722-4040. Sponsor: 3M Company Sponsor Study ID# P3025 2 ASciCorpomlionlAScl-DuluLh EnvironmentalTestingDivision ASci Study [D# 5010-068 2.0OBJECTIVE(S) To determinethe4-dmedianeffecltoading(EL50)and the4-d no observableeffecltoading (NOEL) of thetestsubstance,- ,.;),Lo-S forgreenalga(Selenastrum capricomutumu)nderstatitcesctonditions. This stud was conductedaccordingto OECD Guideline201 (OECD 1993). Test solution preparationwas water-accommodated fractio(nWAF) per Girling& Whale 1994. 3.0 TEST METHODS TestSubstance Test Organisms Algal Medium Sponsor: 3M Company Sponsor Study ID# P3025 Properties: two phase liquid,specific gravityca. 1.2 (water= 1),stable,and insolublein water. Testloadings:0 (algalmedium control),51.8,86.4, 144, 240, and 400 mg/L. Test loadingrange was determined from range-findihgtestresults. Each testloadingwas preparedby mass additionof testsubstanceto 500-ml aliquotsof algalmedium. Test substancewas thoroughlyhomogenized beforeeach mass was determined. Each solutionwas vigorously stirred (vortex1/2 of flasksd'epth)for21 hours to obtainWAF'S. Test solution obtainedfrom flasksby pipettingfrom mid-depth. Source: ATCC 22662 Inoculum preparation: Algal stock was subculturedin OECD recommended medium for4 days beforetestinitiation. Use in test:Stock culturewas dilutedand spikedintotestsolutionsto obtainan initiaclelldensityof 1.0 X 104 cellsper ml (1.2 X 104 actual concentration). Preparation:Deionizedwaterwas spikedwith nutrientstocks(including Na2EDTA) to OECD recommended concentrationsT.he most recent chemical analysisfordeionizedwater isin Raw Data Package. CharacteristicsA:t testtime,the medium had a pH of 7.95. Sterilizevdia 0.22 filtratiporniorto use in the test. 3 Exposure Chambers Incubation Observations Endpoint [-Calculations ASci Corporation/ASci-Duluth EnvironmentaTlestingDivision AScl StudyID# 5Olo-o6a 7@pe: Sterilize2d50-ml borosilicate-glaEsrslenmeyer flaskssealedwith foam stoppers. Use in test: 100 ml of algalmedium or testsolution/chamber.Six control replicateasnd threereplicatepser testsubstanceloadingwere employed. During the test,keptstoppered,exceptwhen experimentalobservations were made. Duration: 4 days. Daityphotoperiod:Continuousat740 ft-candleussingcool-white fluorescenltamps. Test chambers heldon shakertablesetat 100 rpm. Temperature:25.0 2*C. Biological:Daily cellcounts usingdirectmicroscopicenumerationvia Improved Neubauer hemocytometer. Water chemistry:(1)At testinitiatioand terminationp-H, conductivity, and temperature(,2)daily-temperature.All determinationwsere made accordingtoAPHA methods (1995). 1-,2-,3-,and 4-d ELSO'S,and 4-d NOEL based on both cellcountsand averagespecifigcrowth rates(,u). Sponsor: 3M Company Sponsor Study [D# P3025 4 AScl Corpomtion/AScl-Duluth Environmental TestingDivision ASci Study ID# 5010-068 4.0 RESULTS Test Substanr-Teoxicity(Pertinendtataare inTables 1 2, and 3) Test Conditions CellCount 1-d EL50: 55.8 mg/L (50.2-621.). , 1-d ELSO: 57.7 mg/L (54.2-61.5). CellCount 2-d EL50: < 51.8 mg/L. .U2-d ELSO: 54.8 mg/L (50.4-59.6). CellCount 3-d ELso: < 51.8 mg/L. ,u3-d EL50: 61.4mg/L (58.2-64.9). CellCount 4-d ELSO: < 51.8 mg/L. A 4-d EL50: 64.6mg/L (62.5-66.9). CellCount 4-d NOEL: 7.7 mg/L (7.5-81.). pL4-d NOEL: 29.3 mg/L (27.7-31.6). NOEL valuesare approximationsusingthe ICIO as theendpoint. Effectof testsubstancewas determinedto be algistatibcased on the resultsof a postdefinitivteestexposure. Temperature('C): 24.6-25.6. Light intensitfyt-cdls:730-750. pH: 7.78-9.43. I Specificconductivit(ylimhoslcm):134-166. Sponsor: 3M Company Sponsor Study ID# P3025 5 AScl Co rporatino/AScI-Dututh EnvironmentaTlestingDivision AScl Study ID# 5010-068 5.0TEST QA/QC QA/QC Criteria Algalcountsin controlmust increaseby at leasta factorof 16 duringthefirs3t-dof the exposure. Testdurationmust be 4-d. Data During thefirs3t-dof thetestcontrolalgal countsincreasedby a factorof approximately 191. The testdurationwas 4-d. 6.0 REFERENCES American PublicHealthAssociation(APHA). 1995.StandardMethods fortheExaminationof Water and Wastewater.APHA, Washington,D.C. GirlingA,.B., G.F. Whale,and D.M.M. Adema. 1994.A GuidelinSeupplementfor DeterminingtheAquaticToxicityof PoorlyWater-SolublCeomplex MixturesUsing WaterAccommodated FractionsC.hemosphere 29(12):2645-2649. OrganizationforEconomic Cooperationand Development(OECD). 1993.OECD Guidelinefsor Testingof Chemicals.OECD PublicatioInnformatioCnenter,Washington,D.C. Hamilton,M.A., R.C. Russo'a,nd 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 forSublethaTloxicityT:he Inhibition oncent tion(lCp)Approach(Version2.0).EnvironmentaRlesearchLaboratoryD,uluth, Minnesota. Sponsor: 3M Company Sponsor Study ID# P30"'i 6 ASet Corporation/ASci-Duluth EnvironmentalTestingDivision AScl Study ID# 5010-06& TABLE 1. ELSO'S, 95% ConridenceIntervalsa,nd 4-d NOEL'S from AlgalGrowth Inhibition Test with po inL CellCount 1-d EL50 2-d ELSO 3-d ELSO 4-d EL50 4-d NOEL 55.8 <51.8 <51.8 <51.8 7.7 @95% Interva(lm 50.2-62.1 Not Calculable Not Calculable Not Calculable 7.5-8.1 End;po= Average Specific Growth Rate (,u) 'T 1-d F-L50 2-d EL50 3-d ELSO 4-d ELSO .4-d NOEL& (;ngfL) 57.7 54.8 61.4 64.6 29.3 NOEL valuesareapproximatioasusingthe ICIO as theendpoint. 95% Conridence Interva(liiig/L) 54.2-61.5 50.4-59.6 58.2-64.9 62.5-66.9 27.7-31.6 Sponsor: 3M Company SponsorStudyID# P3025 7 ASel Corporation/ASci-Duluth EnvironmentalTestingDivision ASci Study [D# 5010-068 TABLE 2. Cellsper ml and PercentageDifferencefrom Mean ControlValue forSelenastrum capricomutumExposed to P@,cA Too;: 9 Control 51.8 96.4 144 240 4 Repticate@,@ 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 Id 5 7 5 5 6 6.0(-) 3 5 2 3.3 (-44)& 0 0 0.3(-94) 2 0 1 1.0(-93) 1 2 1 1.3(-78) 0 0 0 0(-100) Ce]Wwl (104) .@'2d, 38 36 37 34 33 36 35.7(-) 10 7 7 8.0(-78) 1 2 1.3(-96) 2 1 0 1.0(-97) 1 1 0 0.7(-98) 1 0 0 0.3 (-99) .3 d 200 232 256 232 200 256 229(-) 59 42 41 47.3 (-79) 2 1 3 2.0(-99) 1 1 3 1.7(-99) 1 2 0 1.0(-100) 1 0 0 0.3 (-100) 4d 585 615 675 570 600 630 613 199 222 192 204(-67) 3 1 2 2.0(-100) 0 1 2 1.0(-100) 1 2 2 1.7(-100) 1 0 1 0.7(-100) ParentheticValues % Differencefrom ControlMean V alue 0-d Mean Algal Count = 1.2 X lQ4 Sponsor: 3M Company Sponsor Study ID# P3025 8 ASci Corporation/ASci-Duluth EnvironmentalTestingDivision ASci Study ID# 5010-06g TABLE 3. Average SpecificGrowth Rates (u)and Percentage Differencefrom Mean Control Value for Seknastrum capticornutumExposed to e3D@@ .:Test gdtn,g,. Control 51.8 86.4 144 240 400 R fciatc 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 i 1.897 1.427 1.764 1.427 1.427 1.609 1.609(-) 0.916 1.427 0.511 1.012(-37)b -0.182 0 0 -1.396(-100) 0.511 0 -0.182 -0.182(-100) -0.182 0.511 -0.182 0.080(-95) 0 0 0 0.000 (-IDO) A4trigeS,pmificG,iovAh Rate(u)g ..,::: .. 3 d, 1.728 1.701 1.714 1.672 1.657 1.701 1.696(-) 1.705 1.755 1.788 1.755 1.705 1.789 1.751 1.060 0.882 0.882 0.949(-44) 1.298 1.195 1.177 1.225(-30) -0.091 -0.091 0.255 0.040(-98) 0.255 -0.091 0 -0.091(-100) -0.()91 -0.091 0 -0.269(-100) 0.170 -0.061 0.305 0.170(-90) -0.061 -0.061 0.305 0.116(-93) . -0 061 0.170 0 -0.061(-100) -0.091 0 0 -0.693(-100) -0.061 0 0 -0.462(-100) 4d 1.547 1.560 1.583 1.541 1.554 1.566 1.559 1.276 1.305 1.269 1.284(-18) 0.229 -0.046 0.128 0.128 (-92) 0 -0.046 0.128 -0.046(-100) -0.046 0.129 0.129 0.097 (-94) -0.046 0 -0.046 -0.135(-100) u In CeUs t)erml In Cells/ml-to tn to b ParenthetiVcalues = % Differencefrom Contra[Mean Value 0-d Mean AlgalCount = 1.2 X 104 Sponsor: 3M Company Sponsor Study ID# P3025 9