NREL-Determination-of-Structural-Carbohydrates-and

NRELisanationallaboratoryoftheU.S.DepartmentofEnergy,OfficeofEnergyEfficiency&RenewableEnergy,operatedbytheAllianceforSustainableEnergy,LLC.ContractNo.DE-AC36-08GO28308DeterminationofStructuralCarbohydratesandLignininBiomassLaboratoryAnalyticalProcedure(LAP)IssueDate:April2008RevisionDate:August2012(Version08-03-2012)A.Sluiter,B.Hames,R.Ruiz,C.Scarlata,J.Sluiter,D.Templeton,andD.CrockerTechnicalReportNREL/TP-510-42618RevisedAugust2012NRELisanationallaboratoryoftheU.S.DepartmentofEnergy,OfficeofEnergyEfficiency&RenewableEnergy,operatedbytheAllianceforSustainableEnergy,LLC.NationalRenewableEnergyLaboratory15013DenverWestParkwayGolden,Colorado80401303-275-3000•(LAP)IssueDate:April2008RevisionDate:August2012(Version08-03-2012)A.Sluiter,B.Hames,R.Ruiz,C.Scarlata,J.Sluiter,D.Templeton,andD.CrockerNRELLaboratoryAnalyticalProceduresforstandardbiomassanalysisareavailableelectronicallyat(“Methods”)areprovidedbytheNationalRenewableEnergyLaboratory(“NREL”),whichisoperatedbytheAllianceforSustainableEnergy,LLC(“ASE”)fortheDepartmentOfEnergy.AccesstoanduseoftheseMethodsshallimposethefollowingobligationsontheuser.Theuserisgrantedtheright,withoutanyfeeorcost,touse,copy,modify,alter,enhanceanddistributetheseMethodsforanypurposewhatsoever,exceptcommercialsales,providedthatthisentirenoticeappearsinallcopiesoftheMethods.Further,theuseragreestocreditNREL/ASEinanypublicationsthatresultfromtheuseoftheseMethods.ThenamesNREL/ASE,however,maynotbeusedinanyadvertisingorpublicitytoendorseorpromoteanyproductsorcommercialentityunlessspecificwrittenpermissionisobtainedfromNREL/ASE.TheuseralsounderstandsthatNREL/ASEisnotobligatedtoprovidetheuserwithanysupport,consulting,trainingorassistanceofanykindwithregardtotheuseoftheseMethodsortoprovidetheuserwithanyupdates,revisionsornewversions.THESEMETHODSAREPROVIDEDBYNREL/ASEASISANDANYEXPRESSORIMPLIEDWARRANTIES,INCLUDINGBUTNOTLIMITEDTO,THEIMPLIEDWARRANTIESOFMERCHANTABILITYANDFITNESSFORAPARTICULARPURPOSEAREDISCLAIMED.INNOEVENTSHALLNREL/MRIBELIABLEFORANYSPECIAL,INDIRECTORCONSEQUENTIALDAMAGESORANYDAMAGESWHATSOEVER,INCLUDINGBUTNOTLIMITEDTOCLAIMSASSOCIATEDWITHTHELOSSOFDATAORPROFITS,WHICHMAYRESULTFROMANACTIONINCONTRACT,NEGLIGENCEOROTHERTORTIOUSCLAIMTHATARISESOUTOFORINCONNECTIONWITHTHEACCESS,USEORPERFORMANCEOFTHESEMETHODS.CoverPhotos:(lefttoright)PIX16416,PIX17423,PIX16560,PIX17613,PIX17436,PIX17721Printedonpapercontainingatleast50%wastepaper,including10%postconsumerwaste.1ProcedureTitle:DeterminationofStructuralCarbohydratesandLignininBiomassLaboratoryAnalyticalProcedure1.Introduction1.1Carbohydratesandligninmakeupamajorportionofbiomasssamples.Theseconstituentsmustbemeasuredaspartofacomprehensivebiomassanalysis.Carbohydratescanbestructuralornon-structural.Structuralcarbohydratesareboundinthematrixofthebiomass,whilenon-structuralcarbohydratescanberemovedusingextractionorwashingsteps.Ligninisacomplexphenolicpolymer.1.2PortionsofthisprocedurearesubstantiallysimilartoASTME1758-01“StandardMethodfortheDeterminationofCarbohydratesbyHPLC.”1.3Thisprocedureissuitableforsamplesthatdonotcontainextractives.Thisprocedureusesatwo-stepacidhydrolysistofractionatethebiomassintoformsthataremoreeasilyquantified.Theligninfractionatesintoacidinsolublematerialandacidsolublematerial.Theacidinsolublematerialmayalsoincludeashandprotein,whichmustbeaccountedforduringgravimetricanalysis.TheacidsolubleligninismeasuredbyUV-Visspectroscopy.Duringhydrolysisthepolymericcarbohydratesarehydrolyzedintothemonomericforms,whicharesolubleinthehydrolysisliquid.TheyarethenmeasuredbyHPLC.Proteinmayalsopartitionintotheliquidfraction.Ameasureofacetylcontentisnecessaryforbiomasscontaininghemicellulosewithaxylanbackbone,butnotbiomasscontainingamannanbackbone.AcetateismeasuredbyHPLC.2.Scope2.1Thisprocedureisappropriateforextractivesfreebiomass,whichincludesbiomassthathasbeenextractedusingLAP“DeterminationofExtractivesinBiomass,”aswellasprocesssolidscontainingnoextractives.Resultsarereportedonanovendryweightbasis.Resultsmaybereportedonanasreceivedbiomassbasisoranextractivesfreebasis,dependingontypeofbiomassused.LAP“PreparationofSamplesforBiomassCompositionalAnalysis”shouldbeusedpriortothisprocedure.2.2Thisprocedureisappropriateforbiomasscontainingthecomponentslistedthroughouttheprocedure.Anybiomasscontainingotherinterferingcomponentsnotlistedmustbefurtherinvestigated.2.3Ameasureofacetylcontentisnecessaryforbiomasscontaininghemicellulosewithaxylanbackbone,butnotbiomasscontainingamannanbackbone.2.4AllanalysesshouldbeperformedinaccordancewithanappropriatelaboratoryspecificQualityAssurancePlan(QAP).3.Terminology3.1Ovendryweight(ODW)—theweightofbiomassmathematicallycorrectedfortheamountof2moisturepresentinthesampleatthetimeofweighing.3.2Preparedbiomass—biomasspreparedaccordingtoLAP“Prepar