【astm標準】d 6691 - 09

1、Designation:D6691?09StardTestMethodfDeterminingAerobicBiodegradationofPlasticMaterialsintheMarineEnvironmentbyaDefinedMicrobialConstiumNaturalSeaWaterInoculum1ThisstardisissuedunderthefixeddesignationD6691thenumberimmedi

2、atelyfollowingthedesignationindicatestheyearofiginaladoptioninthecaseofrevisiontheyearoflastrevision.Anumberinparenthesesindicatestheyearoflastreapproval.Asuperepsilon()indicatesaneditialchangesincethelastrevisionreappro

3、val.1.Scope1.1Thistestmethodisusedtodeterminethedegreerateofaerobicbiodegradationofplasticmaterials(includingfmulationadditives)exposedtopregrownpopulationofatleasttenaerobicmarinemicroganismsofknowngeneratheindigenouspo

4、pulationexistinginnaturalseawater.Thetestmethodisconductedundercontrolledlabatyconditions.1.2Thistestmethodisdesignedtoindexpolymermaterialsthatarepossiblybiodegradablerelativetoapositivereferencematerialinanaerobicenvir

5、onment.1.3Thistestmethodisapplicabletoallpolymermaterialscontainingatleast20%carbonthatarenotinhibitytothemicroganismspresentinamarineenvironment.1.4ThevaluesstatedinSIunitsaretoberegardedasthestard.1.5Thereisnosimilareq

6、uivalentISOstard.1.6Thisstarddoesnotpurpttoaddressallofthesafetyconcernsifanyassociatedwithitsuse.Itistheresponsibilityoftheuserofthisstardtoestablishappropriatesafetyhealthpracticesdeterminetheapplicabilityofregulatylim

7、itationspritouse.2.ReferencedDocuments2.1ASTMStards:2D618PracticefConditioningPlasticsfTestingD883TerminologyRelatingtoPlasticsD1193SpecificationfReagentWaterD2593TestMethodfButadienePurityHydrocarbonImpuritiesbyGasChrom

8、atographyD4129TestMethodfTotalganicCarboninWaterbyHighTemperatureOxidationbyCoulometricDetection3.Terminology3.1DefinitionsofTermsSpecifictoThisStard—DefinitionsoftermsapplyingtothistestmethodappearinTerminologyD883.4.Su

9、mmaryofTestMethod4.1Thistestmethodconsistsofthefollowing:4.1.1ingacterizing(carboncontentmolecularweight)plasticmaterialsftesting4.1.2Preparingaunifminoculumofvariousisolatedmarinemicroganismsobtaininganaturalseawatersam

10、ple(withaddedinganicnutrients)fthetestrelyingonthemicrobespresentastheinoculum.4.1.3Exposingthetestmaterialstotheinoculum4.1.4Usingarespirometertomeasurethetotalbiogas(CO2)producedasafunctionoftime4.1.5Assessingthedegree

11、ofbiodegradability.4.2BiodegradabilityisassessedbydeterminingtheproptionofpolymerCconvertedtobiogasC.Thepercentoftheeticalgasproductionexpressedasafractionofthemeasuredtheeticalcarboncontentofthetestmaterialisreptedasafu

12、nctionoftime.5.SignificanceUse5.1Theuseofplasticsaboardshipsisontherisetheuseoftheseaasatrashdumpingsiteisnolongerapossibilityconsequentlythedisposalofplasticmaterialswhileatsearemainsamajissue.Itispossiblethatbiodegrada

13、bleplasticswillhelptoallaypublicconcernbyallowingfthesafedisposalofplasticmaterialsatsea.Thistestmethodhasbeendevelopedtoassesstheratedegreeofaerobicbiodegradationofplasticsexposedtomarinemicroganisms.Aerobicbiodegradati

14、onisdeterminedbymeasuringtheamountofbiogas(carbondioxide)producedduringsuchanexposure.5.2Itisacceptabletousethedegreerateofaerobicbiodegradabilityofaplasticundertheconditionsofthistest1Thistestmethodisunderthejurisdictio

15、nofASTMCommitteeD20onPlasticsisthedirectresponsibilityofSubcommitteeD20.96onEnvironmentallyDegradablePlasticsBiobasedProducts.CurrenteditionapprovedNov.152009.PublishedDecember2009.iginallyapprovedin2001.Lastpreviousedit

16、ionapprovedin2001asD669101.DOI:10.1520D669109.2FreferencedASTMstardsvisittheASTMwebsitewww.astm.gcontactASTMCustomerServiceat.FAnnualBookofASTMStardsvolumeinfmationrefertothestard’sDocumentSummarypageontheASTMwebsite.Cop

17、yright?ASTMInternational100BarrHarbDrivePOBoxC700WestConshohockenPA194282959.UnitedStates1significanttidalflowacteristicsasitispossiblethatthiswillnotberepresentativeofnaturalseawater.8.Hazards8.1Allmicroganismspresentth

18、epossibilityofdiseaseshallbehledwithduecaution.Hsshallbewashedbefeafterexposure.Latexglovessafetyglassesshallbeusedalongwithamouthcover.Allspillscontainingganismsshallbecleanedwithgermicidalantibacterialagentsalloldcultu

19、resshallbeautoclavedbefebeingdiscarded.8.2Thistestmethodrequirestheuseofhazardouschemicals.Avoidcontactwithchemicalsfollowthemanufacturer’sinstructionsMaterialSafetyDataSheets.8.3Allpurchasedmediaalsocanbehazardous.Reada

20、llsafetyinstructions.9.InoculumTestganisms9.1Theinoculumshallconsistofthecompositionsfoundin9.1.19.1.2.9.1.1Aminimumoftentestganisms.Themicroganismswereidentifiedbyusingbacterialidentificationtests(thatisBiologsystemgram

21、stains).Theiridentificationstoatleastgenusarethefollowing:AlteromonashaloplanktisXanthomonascampestriVibrioalginolyticusVibrioproteolyticusActinomycetesp.BacillusmegateriumBacillussp.Zoostersp.Pseudomonassp.Pseudomonassp

22、hasmultiplespecies.9.1.2Naturalseawatercollectedinthelocalareathatdoesnothaveanyhydrocarbonresiduepresentonthesurfaceincludestheadditionofinganicnutrients(.5gLofNH4Cl.1gLofKH2(PO4)maintainsaerationduringtherespirometry.1

23、0.TestSpecimen10.1Weightestsamplestothenearest0.1mghavesufficientcarboncontent(minimum20%)toyieldcarbondioxidevolumesthatcanbemeasuredaccuratelybytherespirometer.Amethodfdeterminingcarboncontentofthetestmaterialisdetermi

24、nedbycalculationelementalanalysisinaccdancewithTestMethodD4129.10.2Acceptablefmsinwhichitispossiblefthetestspecimentobeincludedarepowdersfilmspiecesfragmentsfmedarticlesaqueoussolutions.Thetestmaterialsshallbeconditioned

25、inaccdancewithPracticeD618.Testspecimensinthefmofpowdersshallbeacterizedastoparticlesizedistribution.Meanparticlesizelessthan25mmisrecommended.10.3Ifthespecimensaresolidsgrindingisrecommendedtoobtainapowderfmofthesamplet

26、omaximizesurfacearea.Cryogenicallymillingwithliquidnitrogenbymeansofimpactisarecommendedmethodtoobtainpowders.10.4Optional—UseTestMethodD2593tomeasurerecdthemolecularweightofthetestmaterial.11.Procedure11.1InoculumBuildu

27、pPreparation:11.1.1Allmediashallbepreparedinaccdancewiththedirectionsdescribedonthelabel.FtymLptionsofthemarinebrothshallbeplacedinuptothirteen125mLErlenmeyerflasks(onefeveryganism)autoclavedf20minat121Cat15lbofpressure.

28、11.1.2Eachflaskshallbeinoculatedwith250Lstockculturesofthemicroganismsthatweregrownovernighttoanopticaldensityofapproximately2.0atawavelengthof660placedinashakerincubatat30C.11.1.3Whenthecultureshavereachedthestateofgrow

29、thwherethecellsareinlatelogarithmicgrowthearlystationaryphase(24h)eachseparateculturemustbecentrifugedatarelativecentrifugalfce(RCF)ofapproximately26890(g)f8mintoobtainpellets.11.1.4Aftercentrifugingdecantthemediaawayfro

30、mthepelletsresuspendthepelletsin20mLofminimalmarinesolution.Repeatthecentrifugingdecantingtoremoveanycarbonsourcefromthemicroganisms.Afterdecantingfthesecondtimeresuspendthepelletsin10mLofminimalmarinesolution.Repeatthec

31、entrifugingdecantingofthesolutionthenresuspendthepelletsin4mLofminimalmarinesolution.11.1.5Placethe4mLresuspensionsofallthemicroganismsintoasinglesterileflaskwithcap.Mixthevariousresuspensionstogetherbytheuseofavtexonthe

32、flask.11.2PreparationofRespirometryFlasks:11.2.1Astocksolutioncontainingenoughminimalmarinesolutionfthenumberofflaskstobeusedshallbeautoclavedat121Cf20minpritothedayoftheexperiment.Atubewithattachedairspargeraswellasallo

33、ftheemptyrespirometerflaskslidsshallalsobeautoclaved.11.2.2Seventy–fivemLoftheminimalmarinestocksolutionnaturalseawaterwithinganicnutrientsshallbeplacedasepticallyintoeachrespirometry125mLbottleinasterileenvironment.11.3

34、TestReference(Control)Specimen:11.3.1Thetestcontrolspecimensmustbesterilizedbefeplacementintotherespirometrybottles.Specimensizeisusually20mgdependingonthecarboncontentofthesample(preweighed60.1mg).Sterilizationofpolymer

35、susuallycannotbeperfmedthroughtheuseofanautoclaveduethelimitedthermalstabilityofthepolymerstheeffectofhightemperaturesonapolymer’sstructure.Fthisreasonethyleneoxidesterilizationofsamplesisrecommended.11.3.2Withinasterile

36、environmentconstitutedbyalaminarflowhoodthesamplesshallbeaddedtotherespirometryflasktheflasklidscrewedon.Thesampledreferencematerialsblanksshallbepreparedintriplicate.11.4InoculatingtheRespirometryFlasks:11.4.1Inoculatee

37、achoftherespirometryflaskscontainingtheminimalmarinestocksolutionwith1006Loftheinoculumestablishedin11.1.5.Ifnaturalseawaterwithinganicnutrientsisusedthennoinoculationofpregrownmicroganismsisdone.11.4.2Reserveoneofthebot