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TOXICITY TO AQUATIC PLANTS
TEST SUB,9-TANCE
Identity:Mixturecontainingperfluorooctanesulfonatwehich may also be referredtoas PFOS, FC-95, or as a component of FC-203FC. (1Octanesulfoniaccid)(CAS # 2795-39-3).
Remarks: The 3M productionlotnumber was notnoted. The testsample isFC-203FC, identifiebdy the laboratorays R1335. Currentinformation indicatesitisa mixtureof0.85% PFOS, 17% diethyleneglycolbutylether, 74.46% water,2.34% acrylifcoamer, 1.7% alkylethersulfate2,.55% sodium octylsulfate0,.12% sodium nitrit0e.,09% sodium molybdate,0.85% triethanolaminea,nd 0.04% tolyltriazole.
The followingsummary applies to a mixture with incompletely characterizedconcentrations of impurities.Data may not accurately retlecttoxicityof the fluorochemical component of the testsample.
MET-IOD:
Method: OECD Guideline201 Type: Acute Static GLP: No Year study performed: 1997 Species: Selenastrumcapricomutum Source: Originalloybtainedfrom ATCC, MD, maintainedatASci-DETD, Duluth,MN. Element basis: Algalcellcounts (cells/mls)p,ecificgrowth rates. Exposure period: 96 hours StatisticaMlethods: Statisticanlalyseswere performed usingthe Trimmed Spearman-Karber method. Analyticalmonitoring: pH, conductivitya,nd temperature
Test Conditions:
Algal NutrientMedium: Nutrientmedium per OECD. This nutrientmedium providedallmineralnutrientesssentialforalgalgrowth and alsoserved as the diluentforallalgaloperations.The pH ofthissyntheticalgalmedium was 8.0. Stock and test solution preparation: A 2000 mg/L primarystock solution was preparedby dissolvin7g00 mg FC-203FC in350 mL algalmedium, followedby mechanical stirringE.ach testconcentrationwas made by volume additionofthestockto algalmedium. Exposure vessels: 250 mi Edenmeyer flaskscontaining100 mi test solutions,ealed withfoam stoppers. Agitation: 100 rpm.
Number ofreplicates3:forexposureconcentratio6nfso,rblankcontrol. Initicaelllloading:1.0 X 104 cells/mL(1.3x 104 cells/mLactual). Number of concentrations:fiveplusa blankcontrol
Lighting: Continuouslightinagt740 ft-candleussingcool-whitefluorescent lamps.
Water Chemistry:
pH range (0-96 hours):
8.00 - 9.90 (controelxposure)
5.20 - 7.83 (1000 mg/L exposure) Conductivityrange (0-96hours):
151 - 186 pmhos/cm (controelxposure)
194 - 205 pmhos/cm (1000 mg/L exposure) Test temperature range (0-96hours):
24.8 - 25.8 *C (incubator)
RESULTS
Nominal concentrations: Blank control,130,216, 360, 600, and 1000 mg/L
Algal Growth Response EC50 Values
Exposure (Contact)
Hours 24 48 72 96
Cell-Count mg/L (95% C.I.) 298(207 -428) <130 (C.I.notcalculable) <130 (C.I.notcalculable) <130 (C.1.notcalculable)
Average Specific Growth Rate
mg/L (95% C.I.) 187 (16 - 209) 138 (Notreliable) 163 (145- 184) 271 (239- 308)
Element values: 96-hourEC50 <130 mg/L (celclount) 96-hour ErC50 271 (239- 308)mg/L (growthrate)
96-hourNOEC = 17 rng/L(celclount) 96-hourNOEC = 57 mg/L (growthrate)
Element valuesare based on nominal concentrations.
Reversibilityof Growth InhibitionA:liquotsofthe 1000 mg/L testsolution
were dilutedwithalgalmedium and culturedforan additionapleriod.Based on growth observed inthe recoveryphase,the effecton algalgrowth was found to be algistatic.
Remarks: Testingwas conducted on the mixtureas describedinthe Test Substance Remarks field.The values reportedapplytothatmixtureand not the fluorochemicalproportionalone.
CONCLUSIONS
FC-203FC exhibitsa 96-hour EC50 cellcount valueof<130 mg/L and a 96hour EIC50 growth ratevalue of271 (239- 308) mg/L.
The 96-hourNo Observed EffectConcentration(NOEC) is17 mg/L forcell count and 57 mg/L forgrowth rate.Thistestsubstance was determined to be algistatic.
Submitter: 3M Company, EnvironmentalLaboratory,P.O. Box 33331, St. Paul,Minnesota, 55133
DATA QUALITY
ReliabilityK:limischranking= 2. This studymeets allthe criterifaorquality testing.However, sample puritywas notpropedy characterizedand the study lacksanalyticaclonfirmatioonfthe amount offluorochemicalproportioninthe solution.
REFERENCES
This testwas conducted by ASci Corporation,DuluthEnvironmentalTesting DivisionD,uluth,MN atthe requestofthe 3M Company, Lab Request number R1335,1997.
OTHER
Last changed: 6/26/00
ASci Corporation/ASci-Duluth EnvironmentaTlestingDivision
ASci Study ID# 5010-069
STUDY TITLE GROWTH INHIBITION OF R1335 FOR GREEN ALGA (Selenastrumcapricornutum)
DATASTANDARD OECD GUIDELINE 201
STUDY COMPLETED April7, 1997
TESTING FACILITY AScl Corporation/AScl-Duluth EnvironmentalTestingDivision
4444 AirparkBoulevard Duluth,MN 55811
Tel.No. (218)722-4040 Fax No. (218)722-2592
STUDY IDENTIFICATION NUMBERS AScl Study ID# 5010-068
3M Company Lab Request# R1335
Sponsor: 3M Company Sponsor Study [Dg R1335
AScl Corporation/ASci-Duluth EnvironmentaTlestingDivision
AScl StudyID# 50io-o68
1.0 GENERAL INFORMATION
StudyTitle DataStandard Sponsor
Sponsor'sRepresentative
Sponsor's Lab Request_# TestingFacility
PrincipalInvestigator ProjectDirector TestingFacilityDirector DefinitivTeest Dates Data Validationand Report Review
GrowthInhibitioofnR1335 forGreenAlga (Selenastrcuampricornutum).
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.
R1335.
ASel Corporation/AScI-DuluthEnvironmental TestingDivision,4444 Airpark Boulevard, Duluth,MN 55811; Tel.No. (218)722-4040.
Todd McGuire
Joe Amato
Donald Mount
March 25-29, 1997.
Alan M0701
Report Signature-
Retentionof Raw Data and FinalReport Locationof Raw Data and FinalReport
Two yearsfrom the datestudy iscompleted.
AScl CorporationA/S cI-DuluthEnvironmental TestingDivision,4444 AirparkBoulevard, Duluth,MN 55811; Tel.No. (218)722-4040.
Sponsor: 3M Company
Sponsor Study ID# R1335
2
AScl Corporation/ASel-Duluth EnvironmentaTlestingDivision
AScl StudyID# 5010-068
2.0OBJECTIVE(S)
To detennintehe4-dmedianeffecctoncentrat(iEoCnSO)andthe4-dno observabelfefect c,oncenfta(dNoOnEC)ofthetesstubstancRe1,335,forgreenalga(Selenxtrucmapricomutum) understatitcesctonditions.
This studywas conductedaccordingtoOECD Guideline201 (OECD 1993).
3.0 TEST METHODS
Test Substance Propenies: R1335, clearamber liquid,stable,and misciblewith water.
Test Organisms
Testconcentrations:0 (algalmedium control),130, 216, 360, 600, and 1000 mg/L. A stocksolutiownas preparedby mass additiontoalgal medium followedby mechanicalstirringE.ach testconcentratiownas made by volume additionof the stockto algalmedium.
Source: ATCC 22662
AlgalMedium
Inoculwn preparation:Algal stockwas subculturedin OECD recommended medium for4 days beforetestinitiation.
Use intest:Stockculturewas dilutedand spikedintotestsolutiontso
obtainan initiaclelldensityof 1.0 X 104 cellsper mi (1.3X 104 actual
concentration).
-
Preparation:Deionizedwaterwas spikedwithnutriensttocks(including Na2EDTA) toOECD recommended concentrationsT.he most recent chemicalanalysisfordeionizedwaterisin Raw Data Package.
Exposure Chambers
CharacteristicsA:t testtime,themedium had a pH of 8.00. Sterilizevdia 0.22 um filtratiporniortouseinthetest.
Type: Sterilizevdiaautoclave250-ml borosilicate-glaEsrslenmeyer flasks sealedwith foam stoppers.
Use in test:100 ml of algalmedium or testsolution/chamber.Six control
replicatesand threereplicatesper testsubstance concentrationwere
employed. During thetest,keptstoppered,exceptwhen experimental 1observationswere made.
Sponsor: 3M Company
Sponsor Study ID# R1335
3
Incubation
Observations
Endpoint Calculations
ASci Corpomtion/ASci-Duluth Environmental Testing Division
ASC-IStudy ID# Soio-06s
Duration:4 days.
Dailyphotoperiod:Continuousat740 ft-candleusingcool-white fluorescenltamps. Testchambers heldon shakertablesetat 100 rpm.
Temperature: 25.0 2*C. Biological:Daily cellcountsusingdirectmicroscopicenumerationvia Improved Neubauer hemacytometer.
Water chemistry:(1)At testinitiatioannd termination-pH, conductivity, and temperature(,2)daily-temperature.Alldeterminationwsere made accordingto APHA methods(1995). 1-,2-,3-,and 4-d EC50's, and 4-d NOEC based on both cellcountsand averagespecifigcrowtb rates(IA).Statisticaanlalysesperformed using Trimmed Spearman-KarberMethod (ECSO's)and lCp (ICIO).
4.0 RESULTS
TestSubstanceToxicity(Pertinendtataare in Tables 1 2, and 3)
CellCount 1-dEC50: 298 mg/L (207-428).111-dEC50: 187 mg/L (168-209).
CeU Count 2-dEC50: < 130 mg/L. 142-dEC50: 138 mg/L (NotReliable).
CellCount 3-d ECSO: < 130 mg/L. IA 3-d EC50: 163 mg/L (145-184).
CellCouni 4-d ECSO: < 130 mg/L. g 4-dEC50: 271 mg/L (239-308).
CellCount4-d NOEC (ICIO):17 mg/L (17-18). A 4-dNOEC (ICIO):57 mg/L (53-62).
Effectwas determinedto be algistatbiacsed on recoveryof cellspreviouslyexposedto the 1000 mg/L testsubstanceconcentration.
Sponsor: 3M Company
Sponsor Study ID# R1335
4
TestConditions
ASci Corporation/AScl-Duluth EnvironmentaTlestingDivision
AScl StudyID# 5oio-o6s
Temperature(OC): 24.8-25.8. Lightintensilyft-cdl7s4:0. pH: 5.20-9.90. -_jkeeidc conductiyLq-(iimhoslcm)1:49-205.
5.0TEST QA/QC
QA/QC Criteria
Algalcountsincontrolmust increasbey at leasta factorof 16 duringthefirs3t-d of the exposure.
Test durationmust be 4-d.
Data
During the firs3t-d of thetestcontrolalgal countsincreasedby a factorof approximately 157.
The testdurationwas 4-d.
6.0 REFERENCES
American PublicHealthAssociation(APHA). 1995.StandardMethods forthe Examinationof Water and Wastewater.APHA, Washington,D.C.
OrganizationforEconomic Cooperationand Development (OECD). 1993.OECD Guidelinesfor Testingof Chemicals.OECD PublicatioInnformationCenter,Washington,D.C.
Hamilton,M.A., R.C. Russo, and R.V. Thurston,1977.Trimmed Spearman-KarberMethod for EstimatingMedian LethalConcentrationisnToxicityBioassays.Environ.Sci.Technol. 11(7): 714-719;Correction12 (4):417 (1978).
US EPA. 1993.A LinearInterpolatiMoenthod forSublethalToxicity:The Inhibition Concentratio(nlCp)Approach (Version2.0).EnvironmentalResearchLaboratory,Duluth, Minnesota.
Sponxor: 3M Company
Sponsor Study M# R1335
5
ASci Corpomtion/ASci-Duluth Environmental TestingDivision
ASci Study ID# 5010-068
TABLE 1- ECso'S,95% CoaridenceIntervalsa,nd 4-dNOECIS from AlgalGrowth Inhibition Test withR1335
CellCount
Endpoint Average Specific Growth Rate (a)
Time 1-d EC50 2-d EC50 3-d ECSO 4-d EC50 4-d NOEC (1clo)
Time
Data*
1-d EC50 2-d EC50 3-d ECSO 4-d EC50 4-d NOEC (,CIO)
Values Based on NorninalTest Concentrations
(mg/L) 298 < 130 < 130 < 130 17
(mg/L) 187 138 163 271 57
95 % Confidence Interva(lmgtl)
207-428 Not Calculable Not Calculable Not Calculable
17-18
95% Confidence Interva(lmg/L)';
168-209 Not Reliable
145-184 239-308
53-62
Sponsor: 3M Company
Sponsor Study ID# R1335
6
ASe-qCorporation/ASci-Duluth Environmental TestingDivision
ASci Study ID# 5010-069
TABLE 2. Cellsper ml and PercentageDifferencefrom Mean ControlValue for Seknastrum capticomutum Exposed to R1335
Control 130 216 360 600 1000
licaft
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
4 2 3 5 3 5 3.7
3 3 3 3.0(-19)
1 I 2 1.3 (-64)
3 3 I 2.3 (-37)
I I 2 1.3(-64)
2 1 2 1.7(-55)
Cells/ml(104)
2d
3d
35
200
35
215
35
200
40
195
31
205
30
210
34.3
204
8 8 9 8.3 (-76)
2 1 1 1.3 (-96)
1 1 1 1.0(-100)
<1 <1 <1 <I (-100)
33 33 30 32.0(-84)
6 10 6 7.3 (-96)
<1 2 1 1(-Ioo)
<1 <1 <1 <1 (-100)
<1 2 <1 0.7(-98)
<1 <1 <1 <I (-100)
41d
726 666600 627 6166 770044 639638 662
140165 180 162(-76)
58 51 65 58.0(-91)
16 20 15 17.0(-97)
<1 <1 <1 <I (-100)
<1 I 0.3(-100)
ParentheticValues % Differencefrom Control Mean Value 0-d Mean Algal Count 1.3 X 104
Sponsor: 3M Company
Sponsor Study ID# R1335
7
AScl Corpomtion/AScl-Duluth Environmental TestingDivision
ASel Study ID# 5010-068
TABLE 3. Average SPecilriGcrowth Rates (&L)and PercentageDifferencefrom Mean Control Value forSelenastrumcapticornutumExposed to R1335
Control
Id 1.046
Average SpecificGrowth Rate (jL)&
2d
3d
1.636
1.686
4d 1.558
130
0.836(-20)
0.927(-43)
1.068(-37)
1.206(-23)
216
0(-100)
0(-100)
0.575 (-66)
0.950(-39)
360
0.571 (-45)
0(-100)
0(-100)
0.643 (-59)
600 1000
0(-100) 0.268 (-74)
0(-100) 0(-100)
0(-100) 0(-100)
0 100) 0(-100)
IA InCoUs permi t,- In Cells/mtlo tu- to
b ParentheticValues = S Diffemnce from ControlMean Value 0-d Mean AlgalCount 1.3 X 104
Sponsor: 3M Company Sponsor Study [D# R1335