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TheIntegratedTotalDietaryFibertestkitis suitableforthemeasurementandanalysisofIntegratedTotalDietaryFiber.
Updatedtoincluderesistant,branchedmaltodextrinsasacomponentindietaryfiber.
K-INTDF-AnIntegratedProcedure(AOACMethod2009.01&2011.25)forthemeasurementofTotalDietaryFiber(includingresistantstarchandnon-digestIBLeoligosaccharides).ThismethodcombinesthekeyattributesofAOACOfficialMethodsofAnalysis2002.02,985.29,991.43,2001.03.
Specificdietaryfiberfractionsaremeasuredasfollows:
1.Insolubledietaryfiber(IDF),HigherMolecularWeightSolubleDietaryFiber(SDFP)andLowerMolecularWeightSolubleDietaryFiber(SDFS)determination(AOACMethod2011.25)
2.TotalHighMolecularWeightDietaryFiber(HMWDF)andSDFSdetermination(AOACMethod2009.01)
Theenzymesusedinthesemethodsareofveryhighpurity;theyareeffectivelydevoidofcontaminatingenzymesactiveonβ-glucan,pectinandarABInoxylan.
Datacalculatorsarelocatedinthe Documentationtab.
ViewMegazyme’slatestGuideforDietaryFiberAnalysis.
Determinationoftotaldietaryfibreandavailablecarbohydrates:Arapidintegratedprocedurethatsimulatesinvivodigestion.
McCleary,B.V.,Sloane,N.&Draga,A.(2015).Starch/Stärke,67(9-10),860–883.
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ThenewdefinitionofdietaryfibreintroducedbyCodexAlimentariusin2008includesresistantstarchandtheoptiontoincludenon-digestibleoligosaccharides.Implementationofthisdefinitionrequirednewmethodology.AnintegratedtotaldietaryfibremethodwasevaluatedandacceptedbyAOACInternationalandAACCInternational(AOACMethods2009.01and2011.25;AACCMethod32–45.01and32–50.01,andrecentlyadoptedbyCodexAlimentariusasaTypeIMethod.However,inapplicationofthemethodtoadiverserangeoffoodsamplesandparticularlyfoodingredients,somelimitationshavebeenidentified.Oneoftheongoingcriticismsofthismethodwasthatthetimeofincubationwithpancreaticα-amylase/amyloglucosidasemixturewas16 h,whereasthetimeforfoodtotransitthroughthehumansmallintestinewaslikelytobeapproximately4 h.Inthecurrentwork,weuseanincubationtimeof4 h,andhaveevaluatedincubationconditionsthatyieldresistantstarchanddietaryvaluesinlinewithileostomyresultswithinthistimeframe.Problemsassociatedwithproduction,hydrolysisandchromatographyofvariousoligosaccharideshavebeenaddressedresultinginamorerapidprocedurethatisdirectlyapplicabletoallfoodsandfoodingredientscurrentlyavailable.
Measurementofdietaryfibrecomponents:theimportanceofenzymepurity,activityandspecificity.
McCleary,B.V.(2001),“AdvancedDietaryFibreTechnology”,(B.V.McClearyandL.Prosky,Eds.),BlackwellScience,Oxford,U.K.,pp.89-105.
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Interestindietaryfibreisundergoingadramaticrevival,thanksinparttotheintroductionofnewcarbohydratesasdietaryfibrecomponents.Muchemphasisisbeingplacedondetermininghowmuchfibreispresentinafood.Linkingaparticularamountoffibretoaspecifichealthbenefitisnowanimportantareaofresearch.Theterm"dietaryfibre"firstappearedin1953,andreferredtohemicelluloses,cellulosesandlignin(Theandere/tf/.1995).Trowell(1974)recommendedthistermasareplacementforthenolongeracceptableterm"crudefibre".Burkitt(1995)haslikenedtheinterestindietaryfibretothegrowthofariverfromitsfirsttrickletoamightytorrentHeobservesthatdietaryfibre"wasfirstviewedasmerelythelessdigestibleconstituentoffoodwhichexertsalaxativeactionbyirritatingthegut",thusacquiringthedesignation"roughage"-atermlaterreplacedby"crudefibre"andultimatelyby"dietaryfibre".Variousdefinitionsofdietaryfibrehaveappearedovertheyears,partlyduetothevariousconceptsusedinderivingtheterm(i.e.originofmaterial,resistancetodigestion,fermentationinthecolon,etc.),andpartlytothedifficultiesassociatedwithitsmeasurementandlabelling(Mongeauetal.1999).Theprincipalcomponentsofdietaryfibre,astrADItionallyunderstood,arenon-starchpolysaccharides(whichinplantfibreareprincipallyhemicellulosesandcelluloses),andthenon-carbohydratephenoliccomponents,cutin,suberinandwaxes,withwhichtheyareassociatedinnature.In1976,thedefinitionofdietaryfibrewasmodifiedtoincludegumsandsomepecticsubstances,basedontheresistancetodigestionofthesecomponentsintheupperintestinaltract.Forthepurposesoflabelling,Englystetal.(1987)proposedthatdietaryfibrebedefinedas"non-starchpolysaccharides(NSP)inthedietthatarenotdigestedbytheendogenoussecretionsofthehumandigestivetract".MethodswereconcurrentlydevelopedtospecificallymeasureNSP(Englystetal.1994).
Dietaryfiberandavailablecarbohydrates.
McCleary,B.V.&Rossiter,P.C.(2007).“DietaryFiber:AnInternationalPerspectiveforHarmonizationofHealthBenefitsandEnergyValues”,(DennisT.GordonandToshinaoGoda,Eds.),AACCInternational,Inc.,pp.31-59.
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Debatecontinuesonthedefinitionofdietaryfiber(DF),methodsformeasurementofDF,andmethodsformeasurementofthecarbohydratesthatarereadilyhydrolyzedandabsorbedinthehumansmallintestine.HenNEBergandStahmanndevelopedthe"Wende"proximatesystemforanalysisoffoodsin1860,andasetofvaluesobtainedusingthismethodwerepublishedbyAtwaterandBryantin1900.ThismethodisstillinuseintheUSAforthemeasurementoftotalcarbohydrate.Inthisprocedure,totalcarbohydrateismeasuredbydifferenceafterdeductingthemoisture,protein,fatandashfromthetotalweight.Carbohydratecalculatedinthiswaycontainsnotonlysugarandstarch,butalsothe"unavailablecarbohydrate"ofDF.However,thereareanumberofproblemswiththisapproach,asthe"bydifference"figureincludesanumberofnon-carbohydratecomponentssuchaslignin,organicacids,tannins,waxesandsomeMaillardproducts.Inadditiontothiserror,itcombinesalloftheanalyticalerrorsfromtheotheranalyses(FAO1997).AneedforinformationonthecarbohydratecompositionoffoodsfordiabeticspromptedMcCanceandLawrence(1929)toattempttomeasurecarbohydratecom¬positiontogainresultsthatwouldbeofBIOLOGicalsignificance.Theydividedthecarbohydratesinfoodsintotwobroadgroups,"available"and"unavailable".Theavailablecarbohydrates,thatis,sugarplusstarch,weredefinedasthosethataredigestedandabsorbedbymanandareglucogenic.Theunavailablecarbohydratesweredefinedasthosethatarenotdigestedbytheendogenoussecretionsofthehumandigestivetract.Inthemid1920s,McCanceobtainedagrantof£30peryearfromtheMedicalResearchCounciltoanalyserawandcookedfruitsandvegetablesfortotal"availablecarbohydrate";valuesneededforcalculatingdiabeticdiets.
Measuringdietaryfibre.
McCleary,B.V.(1999).TheWorldofIngredients,50-53.
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Interestindietaryfibreisundergoingadramaticrevivalthanksinparttotheintroductionofnewcarbohydratesasdietaryfibrecomponents.Muchemphasisisbeingplacedondetermininghowmuchfibreispresentinafood.Linkingaparticularamountoffibretoaspecifichealthbenefitisnowanimportantareaofresearch.TotalDietaryFibre.Theterm“dietaryfibre”firstappearedin1953andreferredtohemicelluloses,cellulosesandlignin(1).In1974,Trowell(2)recommendedthistermasareplacementforthenolongeracceptableterm“crudefibre”Burkitt(3)haslikenedtheinterestindietaryfibretothegrowthofariverfromitsfirsttrickletoamightytorrent.Heobservesthatdietaryfibre“wasviewedasmerelythelessdigestibleconstituentoffoodwhichexertsalaxativeactionbyirritatingthegut“thusacquiringthedesignation“roughage”atermwhichwaslaterreplacedby“crudefibre”andultimatelyby“dietaryfibre”Variousdefinitionsofdietaryfibrehaveappearedovertheyears,partlyduethevariousconceptsusedinderivingtheterm(i.e.originofmaterial,resistancetodigestion,fermentationinthecolonetc.),andpartlytothedifficultiesassociatedwithitsmeasurementandlabelling(4).Theprinciplecomponentsofdietaryfibre,astraditionallyunderstood,arenon-starchpolysaccharides,whichinplantfibreareprincipallyhemicellulosesandcelluloses,andthenon-carbohydratephenoliccomponents,cutin,suberinandwaxeswithwhichtheyareassociatedinNature.
Enzymepurityandactivityinfibredeterminations.
McCleary,B.V.(1999).CerealFoodsWorld,44(8),590-596.
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Dietaryfiberismainlycomposedofplantcellwallpolysaccharidessuchascellulose,hemicellulose,andpecticsubstances,butitalsoincludesligninandotherminorcomponents(1).Basically,itcoversthepolysaccharidesthatarenothydrolyzedbytheendogenoussecretionsofthehumandigestivetract(2,3).Thisdefinitionhasservedasthetargetforthosedevelopinganalyticalproceduresforthemeasurementofdietaryfiberforqualitycontrolandregulatoryconsiderations(4).Mostproceduresforthemeasurementoftotaldietaryfiber(TDF),orspecificpolysaccharidecomponents,eitherinvolvesomeenzymetreatmentstepsoraremainlyenzyme-based.InthedevelopmentofTDFproceduressuchastheProskymethod(AOACInternational985.29,AACC32—05)(5),theUppsalamethod(AACC32-25)(6),andtheEnglystmethod(7),theaimwastoremovestarchandproteinthroughenzymetreatment,andtomeasuretheresidueasdietaryfiber(afterallowingforresidual,undigestedproteinandash).Dietaryfiberwasmeasuredeithergravimetricallyorbychemicalorinstrumentalprocedures.Manyoftheenzymetreatmentstepsineachofthemethods,particularlytheprosky(5)andtheUppsala(6)methodsareverysimilar.Asanewrangeofcarbohydratesisbeingintroducedaspotentialdietaryfibercomponents,theoriginalassayprocedureswillneedtobereexamined,andinsomecasesslightlymodified,toensureaccurateandquantitativemeasurementofthesecomponentsandofTDF.These“new”dietaryfibercomponentsincluderesistantnondigestibleoligosaccharides;nativeandchemicallymodifiedpolysaccharidesofplantandalgalorigin(galactomannan,chemicallymodifiedcelluloses,andagarsandcarrageenans);andresistantstarch.Tomeasurethesecomponentsaccurately,thepurity,activity,andspecificityoftheenzymesemployedwillbecomemuchmoreimportant.Aparticularexampleofthisisthemesurementoffructan.Thiscarbohydrateconsistsofafractionwithahighdegreeofpolymerization(DP)thatisprecipitatedinthestandardProskymethod(5,8)andalowDPfractionconsequentlyisnotmeasured(9).Resistantstarchposesaparticularproblem.Thiscomponentisonlypartiallyresistanttodegradationbyα-amylase,sothelevelofenzymeusedandtheincubationconditions(timeandtemperature)arecritical.
Importanceofenzymepurityandactivityinthemeasurementoftotaldietaryfibreanddietaryfibrecomponents.
McCleary,B.V.(2000).JournalofAOACInternational,83(4),997-1005.
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Astudywasmadeoftheeffectoftheactivityandpurityofenzymesintheassayoftotaldietaryfiber(AOACMethod985.29)andspecificdietaryfibercomponents:resistantstarch,fructan,andβ-glucan.Inthemeasurementoftotaldietaryfibercontentofresistantstarchsamples,theconcentrationofα-amylaseiscritical;however,variationsinthelevelofamyloglucosidasehavelittleeffect.Contaminationofamyloglucosidasepreparationswithcellulasecanresultinsignificantunderestimationofdietaryfibervaluesforsamplescontainingβ-glucan.Pureβ-glucanandcellulasepurifiedfromAspergillusnigeramyloglucosidasepreparationswereusedtodetermineacceptablecriticallevelsofcontamination.Sucrose,whichinterfereswiththemeasurementofinulinandfructooligosaccharidesinplantmaterialsandfoodproducts,mustberemovedbyhydrolysisofthesucrosetoglucoseandfructosewithaspecificenzyme(sucrase)followedbyborohydridereductionofthefreesugars.Unlikeinvertase,sucrasehasnoactiononlowdegreeofpolymerization(DP)fructooligosaccharides,suchaskestoseorkestotetraose.Fructanishydrolyzedtofructoseandglucosebythecombinedactionofhighlypurifiedexo-andendo-inulinases,andthesesugarsaremeasuredbythep-hydroxybenzoicacidhydrazidereducingsugarmethod.Specificmeasurementofβ-glucanincerealflourandfoodextractsrequirestheuseofhighlypurifiedendo-1,3:1,4β-glucanaseandA.nigerβ-glucosidase.β-glucosidasefromalmondsdoesnotcompletelyhydrolyzemixedlinkageβ-glucooligosaccharidesfrombarleyoroatβ-glucan.Contaminationoftheseenzymeswithstarch,maltosaccharide,orsucrose-hydrolyzingenzymesresultsinproductionoffreeglucosefromasourceotherthanβ-glucan,andthusanoverestimationofβ-glucancontent.Theglucoseoxidaseandperoxidaseusedintheglucosedeterminationreagentmustbeessentiallydevoidofcatalaseandα-andβ-glucosidase.
Twoissuesindietaryfibermeasurement.
McCleary,B.V.(2001).CerealFoodsWorld,46(4),164-165.
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Enzymeactivityandpurityofthesetopics,theeasiesttodealwithistheimportanceofenzymepurityandactivity.Asascientistactivelyinvolvedinpolysaccharideresearchoverthepast25years,Ihavecometoappreciatetheimportanceofenzymepurityandspecificityinpolysaccharidemodificationandmeasurement(7).Thesefactorstranslatedirectlytodietaryfiber(DF)methodology,becausethemajorcomponentsofDFarecarbohydratepolymersandoligomers.ThecommitteereportpublishedintheMarchissueofCerealFOODSWORLDrefersonlytothemethodologyformeasuringenzymepurityandactivity(8)thatleduptheAOACmethod985.29(2).Inthisworkenzymepuritywasgaugedbythelackofhydrolysis(i.e.,completerecovery)ofaparticularDFcomponent(e.g.β-glucan,larchgalactanorcitruspectin).Enzymeactivitywasmeasuredbytheabilitytocompletelyhydrolyzerepresentativestarchandprotein(namelywheatstarchandcasein).Theserequirementsandrestrictionsonenzymepurityandactivitywereadequateatthetimethemethodwasinitiallydevelopedandservedasausefulworkingguide.However,itwasrecognizedthattherewasaneedformorestringentqualitydefinitionsandassayproceduresforenzymesusedinDFmeasurements.
Dietaryfibreanalysis.
McCleary,B.V.(2003).ProceedingsoftheNutritionSociety,62,3-9.
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The"goldstandard"methodforthemeasurementoftotaldietaryfibreisthatoftheAssociationofOfficialAnalyticalChemists(2000;method985.29).Thisprocedurehasbeenmodifiedtoallowmeasurementofsolubleandinsolubledietaryfibre,andbuffersemployedhavebeenimproved.However,therecognitionofthefactthatnon-digestibleoligosaccharidesandresistantstarchalsobehavephysiologicallyasdietaryfibrehasnecessitatedare-examinationofthedefinitionofdietaryfibre,andinturn,are-evaluationofthedietaryfibremethodsoftheAssociationofOfficialAnalyticalChemists.Withthisrealisation,theAmericanAssociationofCerealChemistsappointedascientificreviewcommitteeandchargeditwiththetaskofreviewingand,ifnecessary,updatingthedefinitionofdietaryfibre.Itorganisedvariousworkshopsandacceptedcommentsfrominterestedpartiesworldwidethroughaninteractivewebsite.Morerecently,the(US)FoodandNutritionBoardoftheInstituteofHealth,NationalAcademyofSciences,undertheoversightoftheStandingCommitteeontheScientificEvaluationofDietaryReferenceIntakes,assembledapaneltodevelopaproposeddefinition(s)ofdietaryfibre.Variouselementsofthesedefinitionswereinagreement,butnotall.Whatwasclearfrombothreviewsisthatthereisanimmediateneedtore-evaluatethemethodsthatareusedfordietaryfibremeasurementandtomakeappropriatechangeswhererequired,andtofindnewmethodstofillgaps.Inthispresentation,the"stateoftheart"inmeasurementoftotaldietaryfibreanddietaryfibrecomponentswillbedescribedanddiscussed,togetherwithsuggestionsforfutureresearch.
Measurementofnoveldietaryfibres.
McCleary,B.V.&Rossiter,P.(2004).JournalofAOACInternational,87(3),707-717.
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Withtherecognitionthatresistantstarch(RS)andnondigestibleoligosaccharides(NDO)actphysiologicallyasdietaryfiber(DF),aneedhasdevelopedforspecificandreliableassayproceduresforthesecomponents.TheabilityofAOACDFmethodstoaccuratelymeasureRSisdependentonthenatureoftheRSbeinganalyzed.Ingeneral,NDOarenotmeasuredatallbyAOACDFMethods985.29or991.43,theoneexceptionbeingthehighmolecularweightfractionoffructo-oligosaccharides.ValuesobtainedforRS,ingeneral,arenotingoodagreementwithvaluesobtainedbyinvitroproceduresthatmorecloselyimitatetheinvivosituationinthehumandigestivetract.Consequently,specificmethodsfortheaccuratemeasurementofRSandNDOhavebeendevelopedandvalidatedthroughinterlaboratorystudies.Inthispaper,modificationstoAOACfructanMethod999.03toallowaccuratemeasurementofenzymicallyproducedfructo-oligosaccharidesaredescribed.SuggestedmodificationstoAOACDFmethodstoensurecompleteremovaloffructanandRS,andtosimplifypHadjustmentbeforeamyloglucosidaseaddition,arealsodescribed.
Anintegratedprocedureforthemeasurementoftotaldietaryfibre(includingresistantstarch),non-digestibleoligosaccharidesandavailablecarbohydrates.
McCleary,B.V.(2007).AnalyticalandBioanalyticalChemistry,389(1),291-308.
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Amethodisdescribedforthemeasurementofdietaryfibre,includingresistantstarch(RS),non-digestibleoligosaccharides(NDO)andavailablecarbohydrates.Basically,thesampleisincubatedwithpancreaticα-amylaseandamyloglucosidaseunderconditionsverysimilartothosedescribedinAOACOfficialMethod2002.02(RS).Reactionisterminatedandhighmolecularweightresistantpolysaccharidesareprecipitatedfromsolutionwithalcoholandrecoveredbyfiltration.RecoveryofRS(formostRSsources)isinlinewithpublisheddatafromileostomystudies.Theaqueousethanolextractisconcentrated,desaltedandanalysedforNDObyhigh-performanceliquidchromatographybyamethodsimilartothatdescribedbyOkuma(AOACMethod2001.03),exceptthatforlogisticalreasons,D-sorbitolisusedastheinternalstandardinplaceofglycerol.Availablecarbohydrates,definedasD-glucose,D-fructose,sucrose,theD-glucosecomponentoflactose,maltodextrinsandnon-resistantstarch,aremeasuredasD-glucoseplusD-fructoseinthesampleafterhydrolysisofoligosaccharideswithamixtureofsucrase/maltaseplusβ-galactosidase.
Developmentandevaluationofanintegratedmethodforthemeasurementoftotaldietaryfibre.
McCleary,B.V.,Mills,C.&Draga,A.(2009).QualityAssuranceandSafetyofCrops&Foods,1(4),213–224.
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Anintegratedtotaldietaryfibre(TDF)method,consistentwiththerecentlyacceptedCODEXdefinitionofdietaryfibre,hasbeendeveloped.TheCODEXCommitteeonNutritionandFoodsforSpecialDietaryUses(CCNFSDU)hasbeendeliberatingforthepast8yearsonadefinitionfordietaryfibrethatcorrectlyreflectsthecurrentconsensusthinkingonwhatshouldbeincludedinthisdefinition.Asthisdefinitionwasevolving,itbecameevidenttousthatneitherofthecurrentlyavailablemethodsforTDF(AOACOfficialMethods985.29and991.43),noracombinationoftheseandothermethods,couldmeettheserequirements.Consequently,wedevelopedanintegratedTDFprocedure,basedontheprincipalsofAOACOfficialMethods2002.02,991.43and2001.03,thatiscompliantwiththenewCODEXdefinition.Thisprocedurequantitateshigh-andlow-molecularweightdietaryfibresasdefined,givinganaccurateestimateofresistantstarchandnon-digestibleoligosaccharidesalsoreferredtoaslow-molecularweightsolubledietaryfibre.Inthispaper,themethodisdiscussed,modificationstothemethodtoimprovesimplicityandreproducibilityaredescribed,andtheresultsofthefirstroundsofinterlaboratoryevaluationarereported.
Determinationoftotaldietaryfiber(CODEXdefinition)byenzymatic-gravimetricmethodandliquidchromatography:collaborativestudy.
McCleary,B.V.,DeVries,J.W.,Rader,J.I.,Cohen,G.,Prosky,L.,Mugford,D.C.,Champ,M.&Okuma,K.(2010).JournalofAOACInternational,93(1),221-233.
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Amethodforthedeterminationoftotaldietaryfiber(TDF),asdefinedbytheCODEXAlimentarius,wasvalidatedinfoods.BasedupontheprinciplesofAOACOfficialMethodsSM985.29,991.43,2001.03,and2002.02,themethodquantitateshigh-andlow-molecular-weightdietaryfiber(HMWDFandLMWDF,respectively).In2007,McClearydescribedamethodofextendedenzymaticdigestionat37°CtosimulatehumanintestinaldigestionfollowedbygravimetricisolationandquantitationofHMWDFandtheuseofLCtoquantitatelow-molecular-weightsolubledietaryfiber(LMWSDF).Themethodthusquantitatesthecompleterangeofdietaryfibercomponentsfromresistantstarch(byutilizingthedigestionconditionsofAOACMethod2002.02)todigestionresistantoligosaccharides(byincorporatingthedeionizationandLCproceduresofAOACMethod2001.03).ThemethodwasevaluatedthroughanAOACcollaborativestudy.Eighteenlaboratoriesparticipatedwith16laboratoriesreturningvalidassaydatafor16testportions(eightblindduplicates)consistingofsampleswitharangeoftraditionaldietaryfiber,resistantstarch,andnondigestibleoligosaccharides.Thedietaryfibercontentoftheeighttestpairsrangedfrom11.57to47.83.DigestionofsamplesundertheconditionsofAOACMethod2002.02followedbytheisolationandgravimetricproceduresofAOACMethods985.29and991.43resultsinquantitationofHMWDF.ThefiltratefromthequantitationofHMWDFisconcentrated,deionized,concentratedagain,andanalyzedbyLCtodeterminetheLMWSDF,i.e.,allnondigestibleoligosaccharidesofdegreeofpolymerization3.TDFiscalculatedasthesumofHMWDFandLMWSDF.Repeatabilitystandarddeviations(Sr)rangedfrom0.41to1.43,andreproducibilitystandarddeviations(SR)rangedfrom1.18to5.44.Theseresultsarecomparabletootherofficialdietaryfibermethods,andthemethodisrecommendedforadoptionasOfficialFirstAction.
Determinationofinsoluble,soluble,andtotaldietaryfiber(codexdefinition)byenzymatic-gravimetricmethodandliquidchromatography:CollaborativeStudy.
McCleary,B.V.,DeVries,J.W.,Rader,J.I.,Cohen,G.,Prosky,L.,Mugford,D.C.,Champ,M.&Okuma,K.(2012).JournalofAOACInternational,95(3),824-844.
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Amethodforthedeterminationofinsoluble(IDF),soluble(SDF),andtotaldietaryfiber(TDF),asdefinedbytheCODEXAlimentarius,wasvalidatedinfoods.BasedupontheprinciplesofAOACOfficialMethodsSM985.29,991.43,2001.03,and2002.02,themethodquantitateswater-insolubleandwater-solubledietaryfiber.ThismethodextendsthecapabilitiesofthepreviouslyadoptedAOACOfficialMethod2009.01,TotalDietaryFiberinFoods,Enzymatic-Gravimetric-LiquidChromatographicMethod,applicabletoplantmaterial,foods,andfoodingredientsconsistentwithCODEXDefinition2009,includingnaturallyoccurring,isolated,modified,andsyntheticpolymersmeetingthatdefinition.ThemethodwasevaluatedthroughanAOAC/AACCcollaborativestudy.Twenty-twolaboratoriesparticipated,with19laboratoriesreturningvalidassaydatafor16testportions(eightblindduplicates)consistingofsampleswitharangeoftraditionaldietaryfiber,resistantstarch,andnondigestibleoligosaccharides.Thedietaryfibercontentoftheeighttestpairsrangedfrom10.45to29.90%.DigestionofsamplesundertheconditionsofAOAC2002.02followedbytheisolation,fractionation,andgravimetricproceduresofAOAC985.29(anditsextensions991.42and993.19)and991.43resultsinquantitationofIDFandsolubledietaryfiberthatprecipitates(SDFP).Thefiltratefromthequantitationofwater-alcohol-insolubledietaryfiberisconcentrated,deionized,concentratedagain,andanalyzedbyLCtodeterminetheSDFthatremainssoluble(SDFS),i.e.,alldietaryfiberpolymersofdegreeofpolymerization=3andhigher,consistingprimarily,butnotexclusively,ofoligosaccharides.SDFiscalculatedasthesumofSDFPandSDFS.TDFiscalculatedasthesumofIDFandSDF.Thewithin-laboratoryvariability,repeatabilitySD(Sr),forIDFrangedfrom0.13to0.71,andthebetween-laboratoryvariability,reproducibilitySD(sR),forIDFrangedfrom0.42to2.24.Thewithin-laboratoryvariabilitysrforSDFrangedfrom0.28to1.03,andthebetween-laboratoryvariabilitysRforSDFrangedfrom0.85to1.66.Thewithin-laboratoryvariabilitysrforTDFrangedfrom0.47to1.41,andthebetween-laboratoryvariabilitysRforTDFrangedfrom0.95to3.14.Thisiscomparabletootherofficialandapproveddietaryfibermethods,andthemethodisrecommendedforadoptionasOfficialFirstAction.
MeasurementoftotaldietaryfiberusingAOACmethod2009.01(AACCInternationalapprovedmethod32-45.01):Evaluationandupdates.
McCleary,B.V.,Sloane,N.,Draga,A.&Lazewska,I.(2013).CerealChemistry,90(4),396-414.
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TheCodexCommitteeonMethodsofAnalysisandSamplingrecentlyrecommended14methodsformeasurementofdietaryfiber,eightofthesebeingtypeImethods.OfthesetypeImethods,AACCInternationalApprovedMethod32-45.01(AOACmethod2009.01)istheonlyprocedurethatmeasuresallofthedietaryfibercomponentsasdefinedbyCodexAlimentarius.OthermethodssuchastheProskymethod(AACCIApprovedMethod32-05.01)givesimilaranalyticaldataforthehigh-molecular-weightdietaryfibercontentsoffoodandvegetableproductslowinresistantstarch.Inthecurrentwork,AACCIApprovedMethod32-45.01hasbeenmodifiedtoallowaccuratemeasurementofsampleshighinparticularfructooligosaccharides:forexample,fructotriose,which,intheHPLCsystemused,chromatographsatthesamepointasdisaccharides,meaningthatitiscurrentlynotincludedinthemeasurement.Incubationoftheresistantoligosaccharidesfractionwithsucrase/β-galactosidaseremovesdisaccharidesthatinterferewiththequantitationofthisfraction.Thedietaryfibervalueforresistantstarchtype4(RS4),variessignificantlywithdifferentanalyticalmethods,withmuchlowervaluesbeingobtainedwithAACCIApprovedMethod32-45.01thanwith32-05.01.ThisdifferenceresultsfromthegreatersusceptibilityofRS4tohydrolysisbypancreaticα-amylasethanbybacterialα-amylase,andalsoagreatersusceptibilitytohydrolysisatlowertemperatures.OnhydrolysisofsampleshighinstarchintheassayformatofAACCIApprovedMethod32-45.01(AOACmethod2009.01),resistantmaltodextrinsareproduced.Themajorcomponentisaheptasaccharidethatishighlyresistanttohydrolysisbymostofthestarch-degradingenzymesstudied.However,itishydrolyzedbythemaltase/amyloglucosidase/isomaltaseenzymecomplexpresentinthebrushborderliningofthesmallintestine.Asaconsequence,AOACmethods2009.01and2011.25(AACCIApprovedMethods32-45.01and32-50.01,respectively)mustbeupdatedtoincludeanadditionalincubationwithamyloglucosidasetoremovetheseoligosaccharides.
ModificationtoAOACOfficialMethods2009.01and2011.25toallowforminoroverestimationoflowmolecularweightsolubledietaryfiberinsamplescontainingstarch.
McCleary,B.V.(2014).JournalofAOACInternational,97(3),896-901.
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AOACOfficialMethods2009.01and2011.25havebeenmodifiedtoallowremovalofresistantmaltodextrinsproducedonhydrolysisofvariousstarchesbythecombinationofpancreaticα-amylaseandamyloglucosidase(AMG)usedintheseassayprocedures.Themajorresistantmaltodextrin,63,65-di-α-D-glucosylmaltopentaose,ishighlyresistanttohydrolysisbymicrobialα-glucosidases,isoamylase,pullulanase,pancreatic,bacterialandfungalα-amylaseandAMG.However,thisoligosaccharideishydrolyzedbythemucosalα-glucosidasecomplexofthepigsmallintestine(whichissimilartothehumansmallintestine),andthusmustberemovedintheanalyticalprocedure.HydrolysisoftheseoligosaccharideshasbeenbyincubationwithahighconcentrationofapurifiedAMGat60°C.ThisincubationresultsinnohydrolysisorlossofotherresistantoligosaccharidessuchasFOS,GOS,XOS,resistantmaltodextrins(e.g.,Fibersol2)orpolydextrose.TheeffectofthisadditionalincubationwithAMGonthemeasuredleveloflowmolecularweightsolubledietaryfiber(SDFS)andoftotaldietaryfiberinabroadrangeofsamplesisreported.Resultsfromthisstudydemonstratethattheproposedmodificationcanbeusedwithconfidenceinthemeasurementofdietaryfiber.
DietaryfibrefractionsincerealfoodsmeasuredbyanewintegratedAOACmethod.
Hollmann,J.,Themeier,H.,Neese,U.&Lindhauer,M.G.(2013).Foodchemistry,140(3),586-589.
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Thereliabledeterminationofsoluble,insolubleandtotaldietaryfibreinbakedgoodsandcerealfloursisanimportantissueforresearch,nutritionallabellingandmarketing.Wecomparedtotaldietaryfibre(TDF)contentsofselectedcerealbasedfoodsdeterminedbyAOACMethod991.43andthenewAOACMethod2009.01.Fifteenbreadandbakeryproductswereincludedinthestudy.OurresultsshowedthatTDFvaluesofcerealproductsdeterminedbyAOACMethod2009.01werealwayssignificantlyhigherthanthosedeterminedbyAOACMethod991.43.ThiswasexplainedbytheinclusionoflowmolecularweightsolublefibrefractionsandresistantstarchfractionsintheTDFmeasurementbyAOAC2009.01.ThisdocumentsthatnutritionallabellingofcerealproductsposesthechallengehowtoupdateTDFdatainnutrientdatabasesinareasonabletimewithanacceptableexpenditure.
AdaptationoftheAOAC2011.25IntegratedTotalDietaryFiberAssaytodeterminethedietaryfiberandoligosaccharidecontentofdryediblebeans.
Kleintop,A.E.,Echeverria,D.,Brick,L.A.,Thompson,H.J.&Brick,M.A.(2013).JournalofAgriculturalandFoodChemistry,61(40),9719-9726.
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Dietaryfiber(DF)hasimportanthealthbenefitsinthehumandiet.Developingdryediblebean(PhaseolusvulgarisL.)cultivarswithimprovedDFandreducednondigestibleoligosaccharidecontentisanimportantgoalfordrybeanbreederstoincreaseconsumeracceptance.TodetermineifgeneticvariationexistsamongdrybeancultivarsforDF,twopopulationsofdiversedrybeancultivars/linesthatrepresenttwocentersofdrybeandomesticationwereevaluatedfordietaryfiberusingtheIntegratedTotalDietaryFiberAssay(AOAC2011.25).Thisassaywasadaptedtomeasurewaterinsolubledietaryfiber,watersolubledietaryfiber,oligosaccharidesraffinoseandstachyose,andthecalculatedtotaldietaryfiber(TDF)contentofcookeddrybeanseed.TheAOAC2011.25protocolwasmodifiedbyusingaquick,simple,andsensitivehigh-performanceliquidchromatographymethodpairedwithanelectRochemicaldetectionmethodtoseparateandquantifyspecificoligosaccharides,andusingduplicatesamplesasreplicatestogeneratestatisticalinformation.TheTDFofdrybeanentriesrangedfrom20.0to27.0%inpopulationIandfrom20.6to25.7%inpopulationII.Totaloligosaccharidesrangedfrom2.56to4.65%inpopulationIandfrom2.36to3.84%inpopulationII.Theresultssuggestthatsignificantgeneticvariationexistsamongdrybeancultivars/linestoallowforgeneticselectionforimprovedDFcontentindrybeansandthatthemodificationstotheAOAC2011.25methodweresuitableforestimatingDFincookeddryediblebeans.
ValidationofMethods
AACCMethod32-45.01
AACCMethod32-50.01
AOACMethod2009.01
AOACMethod2011.25
CODEXMethodTypeI
ForthedeterminationofTotalDietaryFiberincerealproducts,foodstuffs,feedsandothermaterials.CODEXTypeImethod(2011)
Principle:
(Pancreaticα-amylase+amyloglucosidase)
(1)Non-resistantstarch+H2O→D-glucose
(protease)
(2)Protein+H2O→peptides
(3)IDF(includingresistantstarch)andalcoholprecipitated
solubleDF(SDFP)determinedgravimetrically
(4)AlcoholsolubleDF(SDFS)determinedbyHPLC
(5)AshandresidualproteindeterminedonDFresidues
andsubtracted
Kitsize: 100assays
Method: Hydrolysis/removalofnon-dietary
fibrecomponents
Totalassaytime: ~3hrwork(over2days)
Detectionlimit: 0.5-100%ofsampleweight
Applicationexamples:
Foodingredients,foodproductsandothermaterials
Methodrecognition:
AOAC(Methods2009.01and2011.25),AACC(Method32-45.01and
32-50.01)andCODEX(TypeImethod)
Advantages
- TheonlymethodthatisconsistentwiththeCODEXAlimentariusdefinitionofdietaryfibre
- Highpurity/standardisedenzymesemployed
- Allreagentsstablefor>2years
- Mega-Calc™softwaretoolisavailablefromourwebsiteforhassle-freerawdataprocessing
- Verycompetitiveprice(costpertest)
品牌介绍
Megazyme品牌产品简介
来源：作者：人气：2149发表时间：2016-05-19 10:59:00【大 中 小】
Megazyme是一家全球性公司，专注于开发和提供用于饮料、谷物、乳制品、食品、饲料、发酵、生物燃料和葡萄酒产业用的分析试剂、酶和检测试剂盒。Megazyme的许多检测试剂盒产品已经为众多官方科学协会（包括AOAC, AACC , RACI, EBC和ICC等），经过严格的审核，批准认证为官方标准方法，确保以准确、可靠、定量和易于使用的测试方法，满足客户的质量诉求。
Megazyme的主要产品线包括：
◆ 检测试剂盒
◆ 酶
◆ 酶底物
◆ 碳水化合物
◆ 化学品/仪器
官网地址：http://www.megazyme.com
检测试剂盒特色产品：
货号
中文品名
用途
K-ACETAF
乙酸[AF法]检测试剂盒
酶法定量分析乙酸最广泛使用的方法
K-ACHDF
可吸收糖/膳食纤维检测试剂盒
酒精沉淀法测定膳食纤维
K-AMIAR
氨快速检测试剂盒
用于包括葡萄汁、葡萄酒以及其它食品饮料样品中氨含量的快速检测分析。
K-AMYL
直链淀粉/支链淀粉检测试剂盒
谷物淀粉和而粉中直链淀粉/支链淀粉比例和含量检测
K-ARAB
阿拉伯聚糖检测试剂盒
果汁浓缩液中阿拉伯聚糖的检测
K-ASNAM
L-天冬酰胺/L-谷氨酰胺和氨快速检测试剂盒
用于食品工业中丙烯酰胺前体、细胞培养基、以及上清液组分中、L-天冬酰胺，谷氨酰胺和氨的检测分析
K-ASPTM
阿斯巴甜检测试剂盒
专业用于测定饮料和食品中阿斯巴甜含量，操作简单
K-BETA3
β-淀粉酶检测试剂盒
适用于麦芽粉中β-淀粉酶的测定
K-BGLU
混合键β-葡聚糖检测试剂盒
测定谷物、荞麦粉、麦汁、啤酒及其它食品中混合键β-葡聚糖(1,3:1,4-β-D-葡聚糖)的含量
K-CERA
α-淀粉酶检测试剂盒
谷物和发酵液（真菌和细菌）中α-淀粉酶的分析测定
K-CITR
柠檬酸检测试剂盒
快速、可靠地检测食品、饮料和其它物料中柠檬酸（柠檬酸盐）含量
K-DLATE
乳酸快速检测试剂盒
快速、特异性检测饮料、肉类、奶制品和其它食品中L-乳酸和D-乳酸(乳酸盐）含量
K-EBHLG
酵母β-葡聚糖酶检测试剂盒
用于测量和分析酵母中1,3:1,6?-β-葡聚糖，也可以检测1,3-葡聚糖
K-ETSULPH
总亚硫酸检测试剂盒
测定葡萄酒、饮料、食品和其他物料中总亚硫酸含量（按二氧化硫计）的一种简单，高效，可靠的酶法检测方法
K-FRGLMQ
D-果糖/D-葡萄糖[MegaQuant法]检测试剂盒
适用于使用megaquant?色度计（505nm下）测定葡萄、葡萄汁和葡萄酒中D-果糖和D-葡萄糖的含量。
K-FRUC
果聚糖检测试剂盒
含有淀粉、蔗糖和其他糖类的植物提取物和食品中果聚糖的含量测定。
K-FRUGL
D-果糖/D-葡萄糖检测试剂盒
对植物和食品中果糖或葡萄糖含量的酶法紫外分光测定。
K-GALM
半乳甘露聚糖检测试剂盒
食品和植物产品中半乳甘露聚糖的含量检测
K-GLUC
D-葡萄糖[GOPOD]检测试剂盒
谷物提取物中D-葡萄糖的含量测定，可以和其它Megazyme检测试剂盒联合使用。
K-GLUHK
D-葡萄糖[HK]检测试剂盒
植物和食品中D-葡萄糖的含量测定，可以和其它Megazyme检测试剂盒联合使用。
K-GLUM
葡甘聚糖检测试剂盒
植物和食品中葡甘聚糖的含量测定。
K-INTDF
总膳食纤维检测试剂盒
总膳食纤维特定检测和分析
K-LACGAR
乳糖/D-半乳糖快速检测试剂盒
用于快速检测食品和植物产品中乳糖、D-半乳糖和L-阿拉伯糖
K-LACSU
乳糖/蔗糖/D-葡萄糖检测试剂盒
混合面粉和其它物料中蔗糖、乳糖和D-葡萄糖的测定
K-LACTUL
乳果糖检测试剂盒
特异性、快速和灵敏测量奶基样品中乳果糖含量
K-MANGL
D-甘露糖/D-果糖/D-葡萄糖检测试剂盒
适合测定植物产品和多糖酸性水解产物中D-甘露糖含量
K-MASUG
麦芽糖/蔗糖/D-葡萄糖检测试剂盒
在植物和食品中麦芽糖，蔗糖和葡萄糖的含量检测
K-PECID
胶质识别检测试剂盒
食品配料中果胶的鉴别
K-PHYT
植酸(总磷)检测试剂盒
食品和饲料样品植酸/总磷含量测量的简便方法。不需要通过阴离子交换色谱对植酸纯化，适合于大量样本分析
K-PYRUV
丙酮酸检测试剂盒
在啤酒、葡萄酒、果汁、食品和体液中丙酮酸分析
K-RAFGA
棉子糖/D-半乳糖检测试剂盒
快速测量植物材料和食品中棉子糖和半乳糖含量
K-RAFGL
棉子糖/蔗糖/D-半乳糖检测试剂盒
分析种子和种子粉中D-葡萄糖、蔗糖、棉子糖、水苏糖和毛蕊花糖含量。通过将棉子糖、水苏糖和毛蕊花糖酶解D-葡萄糖、D-果糖和半乳糖，从而测定葡萄糖含量来确定
K-SDAM
淀粉损伤检测试剂盒
谷物面粉中淀粉损伤的检测和分析
K-SUCGL
蔗糖/D-葡萄糖检测试剂盒
饮料、果汁、蜂蜜和食品中蔗糖和葡萄糖的分析
K-SUFRG
蔗糖/D-果糖/D-葡萄糖检测试剂盒
适用于植物和食品中蔗糖、D-葡萄糖和D-果糖的测定
K-TDFR
总膳食纤维检测试剂盒
总膳食纤维检测
K-TREH
海藻糖检测试剂盒
快速、可靠地检测食品、饮料和其它物料中海藻糖含量
K-URAMR
尿素/氨快速检测试剂盒
适用于水、饮料、乳制品和食品中尿素和氨的快速测定
K-URONIC
D-葡萄糖醛酸/D-半乳糖醛酸检测试剂盒
简单、可靠、精确测定植物提取物、培养基/上清液以及其它物料中六元糖醛酸含量（D-葡萄糖醛酸和D-半乳糖醛酸）
K-XYLOSE
D-木糖检测试剂盒
简单、可靠、精确测定植物提取物、培养基/上清液以及其它物料中D-木糖含量
K-YBGL
Beta葡聚糖[酵母和蘑菇]检测试剂盒
检测酵母和蘑菇制品中1,3:1,6-beta-葡聚糖和α-葡聚糖含量
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