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TOXICITY TO AQUATIC PLANTS TEST SUBSTANCE Identity:A mixturecontainingperfluorooctanesulfonatweh,ich may alsobe referredtoas PFOS, FC-95, oras a component of FC-203CE. (1Octanesulfonicadid)(CAS # 2795-39-3). Remarks: The 3M productionlotnumber was 519.The testsample isFC203CE. Currentinformationindicatesitisa mixtureof 1.84% PFOS, 30% diethyleneglycolbutylether,60.23% water,2.56% Sultone foamer,3.1% sodium octylsulfate2,.1% polyoxyethylenemonooctylphenyl ether,0.12% sodium laurylsulfatea,nd 0.05% tolyltriazole. The followingsummary applies to a mixture with incompletely charactedzed concentrations of impudties. Data may not accurately reflectthe toxicityof the fluorochemical component of the testsample. METHOD: Method: OECD 201 and ISO/DIS 8692 Type: Acute Static GLP: No Year study performed: 1989 Species: Selenastrumcapricomutum Source: Originalloybtainedfrom ATCC, RockvilleM,D, medium at 3M EnvironmentalLaboratory,St.Paul,MN. Element basis: Algalcellcounts (cells/mi). Exposure period: 96 hours StatisticaMlethods: Not given. Analyticalmonitoring: pH and temperature. maintainedinculture Test Conditions: Algal NutrientMedium: Nutrientniedium per OECD 201 and ISO/DIS 8692. This nutrientmedium providedallmineralnutrientesssentialforalgalgrowth and alsoserved as the diluentforallalgaloperations.The pH ofthis syntheticalgalmedium was 7.6adjustedto 8.3. One hundred mL ofthispHad*tstednutrientconcentratesolutionwas added to900 mL ofMilliporMeilliQ water.FinalpH 8.3. Stock and testsolutionpreparation:A 1000 mg/L stocksolutiownas pre ared by dilutin1g000 mg oftestsubstance in900 mL of MilliporMeilliQTRpwater then adding 100 mL ofthe ph-adjustednutrientconcentrate solution.FinalpH ofthisstocksolutionwas 7.9. AJiquotswere then added directltyodilutiownater intestvesselsto createtestsolutions. Exposurevessels2:50mL Erienmeyefrlaskcsontaini1n0g0mL test solutioannd capped withstainlessteelclosures. Agitation:Shaken continuouslayt- 100 rpm. Number of replicates:3 fortheblankcontrolsand 2 fortestmaterial treatments Initiaclellloading: 1.0X 104 cells/mL. Number of concentrations: fiveplus a blankcontrol Lighting:400 foot-candiesfrom continuouscool-whitefluoresoentlighting. Water Chemistry: pH range (0-96hours): 8.3- 9.4(controelxposure) 6.8- 7.9(1000nig/Lexposure) Test temperature(0-96houm): 24 OC (incubator) RESULTS Nominal concentrations:Bk control,100,180,320,560, and 1000 mg/L Algal Growth Response at 96 Hours Nominal Conc., mg/L Bk. Control 100 180 320 560 1000 Avg. Growth Rate 1.3853 1.3398 1.2703 1.0891 1.0129-0.8045 % Change inGrowth Rate -3.3 8.3 21 27 42 Element valuesare based on nominalconcentrations. Revemibilityof Growth Inhibition:Not given. Remarks: Testingwas conductedon a mixtureas describedintheTest Substance Remarks field.The valuesreportedapplytothatmixtureand not the fluorochemicaplroportioanlone. CONCLUSIONS The testsubstanceexhibitas 96-hourpercentchange ingrowthrate compared tothecontrolof-42% atthe 1000 mg/L treatment. Submitter: 3M Company, EnvironmentalLaboratory,P.O. Box 33331, St. Paul,Minnesota,55133 DATA QUALITY Reliability:Klimischranking= 2. This study meets the criterifaorquality testing.However, the study lacksinformationon purityofthetestsubstance and the studylacksanalyticaclonfirmationofthe amount offluorochemical proportioninthe solution. REFERENCES This studywas conducted by the 3M Company, EnvironmentalLaboratory, St.Paul,MN, Lab Request number G 1358-1, 1989. OTHER Last changed: 6/26/00 ENVIRONMENTAL LABORATORY 'y STANDARD NUTRIENT MEDIUM FOR FRESHWATER ALGAE NUTRIENT STOCK SOLUTION CONCENTRATION IN STOCK SOLUTION li MACRONUTRIENTS FINAL CONCENTRATION IN TEST SOLUTION NH Cl mgti 2'6H2o cacl2:2H 20 MGSO 7H 20 KH 2P6 4 STOCK SOLUTION 21 FE-EDTA 1.5 g/L 1.2 g/L 1.8 9/L 1.5 g/L 0.16 g/L 15 mg/L 12 mg/L 18 mg/L 15 .mg/L I 6 mg/L FeCl 6H 0 N'a 2EA;A.3H 20 STOCK SOLUTION 3: TRACE 80 mg/L 100 mg/L ELEMENTS 80 ug/L 100 ug/L H BO MAC,3.4H 0 ZnCl 2 2 COC12.6H 0 CUC12 2H2 0 Na2 ma; 40 iH 20 STOCK SOLUTION NAHCO 3 4: NAHCO 3 185 mg/L 415 mg/L 3 mg/L 1.5 mg/L 0.01 mg/L 7 mg/L 50 g/L 185 ug/L 415 ug/L 3 ug/L 1.5 ug/L 0.01 ug/L 7 ug/L 50 mg/L Sterilize stock solutions 1-4 by membrane filtration (mean pore diameter 0.2 um). SOLUTION 5: NUTRIENT CONCENTRATE (Prepare freaFi-y-bef-oreeach te-s-t.) Add 100 mL of STOCK SOLUTION 1; 10 mL of STOCK SOLUTION 21 10 mL of.STOCK SOLUTION 3; and 10 mL of"STOCK SOLUTION 4. Makeu UP to@.11000@'m'L'with''W, A d st the'-pH'to@-,9. water SOLUTION 6t STANDARD ALGAL NUTRIENT MEDIUM (ue&k+ct- Add 100'mL,0of the H'adj@sted Nutrient Concentrate Solutio to 900 ML f water. REPERENCESs OECDf-198 G@r-o'@wn@.'@ti n n Lb it !'on To a t 2 0 1 n s,;...Gu for@..'iestihg'-of-c-Chomii6Oarlgia,nization, for Ec"Oriomi6 cooi@eration and,'D@Oelopni6nt ;,.P.aris Prance., (2),ISO 1987. Wat'orT, A 1L.'g a 1:',.'Grow'th Inhib @ion ISO/DIS,8692,@@Ints rnAtional .10r;ianlzation for'Standareds izat@io' z Gonevoi -.Sswwitzioirliin i STANDARD ENVIRONMENTAL LABORATORY NUTRIENT MEI)IUM FOR FRESHWATER ALCAE t"72 IUTRIENT STOCK SOLUTION CONCENTRATION IN STOCK SOLUTION 1: MACRONUTRIENT - FINAL CONCENTRATION -IN TEST SOLUTION Nil Cl M961 2 .61120 CaCi 2 .2112 0 MGSO .711 0 KH ps 2 24 STOCK SOL-LITION 2: FE-EDTA 1.5 g/L 1.2.g/L 1.8 g/L 1 5 g/L 0:16 g/L 15 m(u/L 12 mg/L 18 mg/L 15 mg/L 1..6 mg/L STOCK FeCl 6H 0 Na 2 Ea@A.3H 20 SOLUTION 3: TRACE 80 mg/L 100 mg/L ELEMENTS 80 ug/L 100 ug/L H BO MAC,3 2 .4H 2 0 ZnCl 2 coci 2- 6H 2 0 cuci 2H 0 Na 2 mg; 4- 3H 20 STOCK SOLUTIONNAHCO 3 4: NAHCO 3 185 mg/L 415 mg/L 3 mg/L 1.5 mg/L 0.01 mg/L 7 mg/L 50 g/L Sterilize stock solutions 1-4 by membrane (mean pore diameter 0.2 um). 185 ug/L 415 ug/L 3 ug/L 1.5 ug/L 0.01 ug/L. 7 ug/L .50 mg/L filtration SOLUTION 5: NUTRIENT CONCETJTRATE (Prepare freshl bef ,e eazh Ces-t Add 100 mL of STOCK SOLUTION'L 10 mL of STOCK SOLUTION 10 mL of STOCK SOLUTIONt.3 and 10 mL of STOCK SOLUTI( Make up to 1,000 mL with Adjust the pH to 8.3 + water SOLUTION 6: STANDARD A AL"NUTRIE Add 100 'mL of the pH-adjust4d to 900 ML of rr4tA Nuii'i'n '-'Concentr soluti ater.@ REF]ERENCES: 0 0 ECD'.'1984oAlgA'","Growth",@inhi ODS t or,:-,Testin*gof:Cheiiica Igi Organ'i'zatLo 'n ion and.,Developm6nt@,'., P"a*r'*'s@ii@ -@ISO 1987.-:Water Qu'aiity@ ISOIDIS 8692."."I@ternaty'@@ on Go !lovep,-@Switzerl0a'.-, izill or@,-E,*E"onomic ilKt6 iion es 'cor s k)ON STANDARD ENVIRONMENTAL LABORATORY NUTRIENT MEDIUM FOR FRESHWATER ALGAE NUTRIENT CONCENTRATION IN STOCK SOLUTION FINAL CONCENTRATION IN TEST SOLUTION STOCK SOLUTION 1: MACRONUTRIENTS NH Cl P6 t4g8l2- 6H 2 0 CaCl2 2H 20 Mgso :7H 20 KH 2 4 STOCK SOLUTION 2: FE-EDTA 1.5 g/L 1.2 g/L 1.8 g/L 1.5 g/L 0.16 g/L 15 mg/L 12 mg/L 18 mg/L 15 .mg/L 1 6 mg/L FeCl 6H 0 Na 2 E6@A.3H 2 0 STOCK SOLUTION 3: TRACE 80 mg/L 100 mg/L ELEMENTS 80 ug/L 100 ug/L H BO MAC,3 .4H 0 22 ZnCl 2 CoCl .6H 0 2. 2 cucl 2H 0 Na 2M90 4- iH 20 STOCK SOLUTION 4: NAHCO 3 NaHC03 185 mg/L 415 mg/L 3 mg/L 1.5 mg/L 0.01 mg/L 7 mg/L 50 g/L Sterilize stock solutions 1-4 by membrane (mean pore diameter 0.2 um). 185 ug/L 415 ug/L 3 ug/L 1.5 ug/L 0.01 ug/L 7 ug/L 50 mg/L filtration SOLUTION 5: NUTRIENT CONCENTRATE (Prepare freshly before each test.) Add 100 mL of STOCK SOLUTION 1; 10 mL of STOCK SOLUTION 2; 10 mL of STOCK SOLUTION 3; and 10 mL of STOCK SOLUTION 4. Make up to 1,000 mL with Adjust the pH to 8.3 + O.'T. SOLUTION 6: STANDARD ALGAL NUTRIENT =Q water. 'RIO %, k MEDIUM lrczowf,5,40cltsooon/L Add 100 mL oft@e to 900 mL of pu-adjusted Nutrient c OdCW421C6 rate Solution &ter. REFERENCESs (1) OECD 1984. Alga, Growth Inhibition":'Test, 2010 Ins Guidelines for Testing of Chemicals. Organization for.Economic Cooperation and Development, Paris, France. (2) ISO 1967. Water Ouality - Algal.-Growth Inhibition Teste ISO/DIS 8692. International.organization for Standardization, Geneve, Switzerland. EWIRONNINTAL 1ABoRATORy PRESMTER ALGAE GRowni INHIBITION TEST ALGAL CELL COUNR, No./mL HR40CYIUOU MMMD* MATERIAL TEMMAEW, WM Typ /.;p A= SPECIES:Green Al ae, Selenastruncadricornuttn ANALYST- DATE-@:.@@l- TREA7MW T- TOTAL OF 2S FIELDS Comm No./mL MEAN SD pH -,2 1-3 1-4 2--4 14,,F:7(,) /4,1 C-) @loaioo Test Temperattire 0.1 mm de 'a itd of field1 x I and 0.1 om deep. Test methodology tI the: fo Il'o'w-ing two" test protoc0 1s:.. (1) OECD 1201-!'Alga Gi IMibiiien'.--T6st,, (2) US-EPA OBC-$;."'Aii,Acute@,Toxi6ity,Test" _j i@ 10620a0PWO Wple Description 14ATE-RIAL TESTED: ALGAL SPECIES: NT EnvironmentaLlab - Work Shoot AWAL ASMY: WITLC- *FMT (M: BT) CCMPARISON or. GRowni RATES Lab Request No. p.p of Dais: rG Selenaesmit-Lcraupmcraicornutum DAY -DAY to sx (r L; 1.'7 v it)N- (L) SpecificGrowth Rate -In (u) Contro. EWIRO*IWAL LABORATORY FRE-SUATER ALGAE GROMI IMilBITION TEST ALGAL CELL CO(INT,No./mL H50LYrCmh-M MMMD* TEST L MATERLKL TESTM)- 7 TEST TYP - A= SPECIES: Green Algae, Selenastrum canricomutum ANALYST fr DATF,__a:,/a ;NIEMATHwr TOTAL OF 2S FIELDS No./mL MEAN ;t SD Pli UE 17- IC-c ICc,c; I @o Icc,c, 4 -sg lc,ccl,wit <,5CL-iM, 0 5cfccrc 5(d, COC) Z;40100c, zw 000 b.9 oi Test 'Temperatur:e,", 0.1 ma dftp fim@@t ,@rea-o eld: 1 x 1 and 0.1 mm &-ep. T(Ieit'M' theKIblov. C@mfolms'withthe@@.folloW@ng,.tw test protocols: OECD 1201 "AlFa,-Growth.IMibiii'6ri- I @- I -- , -I .- (21 US-EPA OBC-5 -.;,AlgalA.c-u to -- " LobRiquesNto.igzase?-l 9 SunpleNsaiption pap of EnvironmentaLlab - Work Shoot ALM ASSAY: BOTTLE TEST (AA:BT) CCMPARISON OF GROWM RATES Date: MATERIAL TE=: -?) ,/ Analy ALGAL SPECIES: Ereshwater Gr@eenAlizae,Selenastrum capricornutum CELL Cotw GROWRH RATE TREA79W 4-DAY 4-DAY +DAY IC)OK,(QL i- i 2--2- z-4 ]Oc@UlLi I-IS Y, OLO z. in x IOV > 1.1.@kl--i&.2If;,z (a:5 IC) I, ig <I o :E.Yo,lo!5, Ips 51(oA io L.4:x in@s [D A IOS; 1. 2-76 Mq 1,0 1 i.oiz.2 OOLOZ 0 lig4-,@7 n. 2 '-l 4z (1) SpecificGrowth Rate (u) (7) Ra=ent Chanue izar-row:tRhate -in N,% ln NAw t CContrzl-- Samel:- x Control