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BIODEGRADATION TEST SUBSTANCE Identity:Perfluorooctylsulfonadtied,ecyldimethylammoniumsalt;may also be referredto as 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-heptadecaf-luoro-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 rapidlyseparatesafteragitationN.o calculations were made to adjustfortheactualconcentrationofthetestsubstance in the testsample. METHOD: Method: OECD Method 301 D Type: Aerobic GLP: No Year completed: 1997 Contact time:28 days lnoculum: PolyseedC- NX (withnitrificatiinohnibitorl)o,tA306, Polybac Corp, Bethlehem, PA RESULTS COD: 125,000 Mg 02 /kg chemical CBOD Exposure period, Day 5 10 20 28 FirstReplicate CBOD Mean Value (mg/kg) <40,000 <40,000 <40,000 55,039 Second Replicate CBOD Mean Value (mg/kg) <40,000 <40,000 <40,000 <40,000 Remarks: Valuesreporteadreforthetestsample.No calculations were made toadjustforthe concentrationofthe testsubstance inthetest sample. CONCLUSIONS This testingindicatedthatsome portionofthetestsample biodegraded in aerobicenvironmentscapable ofsupportingbiodegradation.This study cannot ruleout thepossibilithyatconditionscould be found thatwould allowforbiodegradationofthe fluorochemicalcomponent, nor could it exclude the possibilitthyata microorganism couldeventuallybe enriched thatcouldmetabolicallaylterthischemical. Submifter: 3M Company, EnvironmentalLaboratory,P.O. Box 33331, St.Paul,Minnesota,55133 DATA QUALITY ReliabilityK:limischranking2. This study lackssufficient charactedzationofthe testsubstance purityand analyticamleasurement oftestconcentrations.Because a mixturewas tested,the testingcannot conclude what portionsofthetestsample were responsibleforthe apparent biodegradationvalues. REFERENCES This studywas conducted by the 3M Company, Environmental Laboratory,St Paul,MN, Lab Request number P3025. OTHER Last changed: 5/24/00 3M ENVIRONMENTAL LABORATORY DETERMINATION OF THE 28-DAY CARBONACEOUS BIOCHEMCIAL OXYGEN DEMAND (CBOD) FOR P3025 OECD Closed BottleTest,#301 D (Modified) STUDY COMPLETED: March 28,1997 FINAL REPORT COMPLETED: May 20,1997 Env. Lab Request Number P3025 Prepared by: Name: Title: Laboratory: Address: Susan A. Beach Advanced Environmental Biologist 3M Environmental Laboratory Building2-3E-09 935 Bush Avenue St Paul,MN 55144 28-Day CBOD Page 1 May 20, 1997 SUMMARY InthisStudy the aerobic,aquaticready biodegradabilitoyf P3025 ingeneralaccordance withthe "Closed Bottle Test",OECD Method No. 301 D was determined. The testsubstance was found tobe not readilybiodegradable, withremoval valuesof <32% and 44% (duplicateo)btainedafter28 days. ORJECTIVE The purpose ofthisstudywas toevaluatethe ready biodegradabilitoyf P3025 after5, 10,20, and 28 days of exposure using a modifiedform of OECD Method 301 D. MATERIALS AND METHODS Tgst Substance: P3025, date 10/7/96,Lot 1 Appearance Aqueous Solubility Two-phase dispersion(clearliquiwdithopaque solid) Apparentlynot completelysolubleinwater attestedconcentrations. AvailablePhysicochemical Properties Date Received Storage BoilingPoint: Vapor Pressure: Vapor Density: EvaporationRate: Solubilitiynwater: SpecificGravity: Percent Volatile: pH:. 1000C ca.18 mm Hg P <1 <1 Insoluble ca.1.2 60-70% 6-8 200C 10/16/96 Inthe dark atambientroom temperature(22 20C) Test Solutions Reported values Test solutionwsere preparedas Water-Accommodated Fractions(WAFs). Four stocksolutionse,ach induplicateo,fapproximately6, 12,25, and 50 nig/Lwere prepared inaeratedmineralnutrienmtedia. Each was capped and allowedto mix overnight,inthe dark,atambient room temperature.Test substance was apparentlynot completelymisciblewithwater. A few whiteparticulatewsere observed ineach stocksolutionaftermixing.The clearliquidportionwas withdrawn forbiodegradationtesting. The reportedvalueswere based on nominal concentrationsinthe test media at the beginningofthe toxicittyest. 28-Day CBOD Page 2 May 20, 1997 Test Orc3anismse Type Source Lot number Preparation Polyseed@-NX (withnitrificatinohnibitor) Poiybac Corp.,Bethlehem, PA A306 Contents ofone Polyseecl&NX capsule placed into500 mL aerated mineralmedia. Solutionstirrefdorsixtyminutes on magnetic mixer. Solutionkeptstirrinwghilealiquotswithdrawn toinoculatetestbottles. Test Condit6ons: Photoperbd Test Vessels TestVokinie Loacfing No. Replicates MineralMedium 24-hours dark 300 mL clearglass Wheaton BOD boftleswithground glassstoppers sealedwitha water sealand capped. 300 mL 5 mL reconstituteidnoculum added to each bottleexcept formedia blanks which receivednone 2 each formedium blank,seeded blank,referencestandard and each testconcentration.Only one replicatfeortoxicitcyheck. Prepared withHach BOD NutrientBufferPillows(catalogue# 14863-98) each dissolvedin19 literMsilliporMeilli-wQater.Glass carboys aerated withlaboratoryairfilteretdhrough a 0.2gm filtefrorapproximately2hours. Carboy openings covered withnew foam plugs and allowedto stand overnightinthedark atambient room temperature. InitiaSltock SolutionLoadings -nominal FirstReplicate 6.7 mg/L 12.3 mg/L 26.9 mg/L 51.6 mg/L Duplicate 5.6 mg/L 10.9 mg/L 24.3 mg/L 54.2 mg/L Test Bottles Each testBOD bottlewas seeded and fillecdompletelyfulwlithsolution from the appropriatewater-accomniodated fraction. Reference Standard Glucose/glutamicacidmixture.Hach BOD standardsolutionampules, cat.# 14865-10. Added 1.5mL ofsolutiontoeach referencestandard BOD bottle. ToxicityCheck Added 1.5mL of Hach glucose/glutamicacidmixtureto a seeded SOD bottlecontainingthe testsubstance P3025 atthe highestloading. 28-Day GSOD Page 3 May 20.1997 TestTerTp.,OC Stock SolutionspH Exposure Period Measuremerts Date Test Initiated Date Test Terminated Procedure 21 -220C, continuouslyrecorded. InitipaHl values ranged from 6.5to6.7. 28-days Dissolvedoxygen at0,5, 10,20,and 28 days. Dissolvedoxygen meter calibratebdy means ofWinklertitratiofnaerated Milli-wOater each day. 2/28/97 3/28/97 The testgenerallyfollowedthe recommendations ofthe OECD GuidelinesforTestingof Chemicals CALCULATIONS The reportedvalues are based on nominalstocksolutionloadingsatthe beginningof the study. The concentrationstested and the correspondingresponse data derivedfrom theduplicatedtestare used to calculate,N possible the mg of carbonaceous biochemical oxygen demand per kg of testsubstance (BOD). The percent biodegradabilityisthen determined by dividingthe CBOD by the Chemical Oxygen Demand (COD) and multiplyingby one hundred. RESULTS Percent biodegradabilityvalues obtained after5, 10, 20, and 28 days of exposure to P3025 are summarized in Table 1. Induplicated measurements, oxygen demand. the percent biodegradabilityof P3025 was reported as <32% and 440/oof chemical ACCEPTABILTTY CRITERIA This biodegradation test meets qualitycriteriparovided intestguidelines: 1) The glucose-glutamic acid reference standard 5-day C-BOD resultswere withinthe range normally seen in thislaboratory.See Figure 1. 2) The oxygen depletion of the medium blanks was < 0.5 mg/L after5-days. 3) The oxygen depletion of the medium blanks was:s 1.5 after28-days. 4) The C-BOD resultforthe toxicitycheck (highesttested concentration plus glucose-glutamic acid)was greater than the value found forglucose-glutamic acid alone. This indicatedthe testsubstance did not inhibitthe microorganisms. 5) Only those data pointswhere the corrected dissolved oxygen drop was > 2.0 mg/L and the final dissolved oxygen concentration inthe bottlewas > 0.5 mg/L were used to calculateC-BOD. 28-Day CBOD Page 4 May 20, 1997 Thistestwas conducted by Trudy K. Stietenhoeteras requested by Pam Wolf of3M SpecialtyChemicals Division. REFERENCES APHA, AWWA, WEF. 1995. Standard Methods forthe ExaminationofWater and Wastewater. Method 5210 B, 5-Day BOD Test. APHA, AWWA, WEF. 1995. Standard Methods forthe ExaminationofWater and Wastewater. Method 4500-0 C. Oxygen (Dissolved)A,zide Modification. OECD. 1993. 301 D Closed BottleTest. GuidelinesforTestingofChemicals. OrganisationforEconomic Cooperationand Development, Paris,France. OECD. 1981. PrincipleosfGood LaboratoryPracticeA,nnex 2,C(81)30 (Final)7:-28. OECD Guidelinesfor Testingof Chemicals. OrganisationforEconomic Cooperation and Development, Paris,France. Polybac. 1997. PolyseecIO-NX,'The Original"CBOD Seed Inoculum. Polybac Corporation,Bethlehem, PA. 28-Day CBOD Page 5 May 20,1997 TABLE 1 SUMMARY OF READY BIODEGRADABILITY OF P3025 Exposure Period 5-Days Duplicate 10-Days Duplicate 20-Days Duplicate 28-Days Dul2licate CBOD, mg0@g <40,000 <40,000 <40,000 <40,000 <40,000 <40,000 55,039 <40,000 *COD reported as 125,000 mg/kg Percent of COD* <32 <32 <32 <32 <32 <32 44 <32 28-Day CBOD Page 6 May 20, 1997 400 . ....................... FIGURE I CBOD GGA 5-daY Reference Standard 350 300 250 200-E 150 -- 100 -- 50-- Results mg./i --it-Average * Lower limitAver.-2std dev * Upper limitaver.+2 std dev 0 11/22/96 12/20/96 Date 1/24/97 3/28197