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.LinktoArticleReadAbstractThenewdefinitionofdietaryfibreintroducedbyCodexAlimentariusin2008includesresistantstarchandtheoptiontoincludenon-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.LinktoArticleReadAbstractInterestindietaryfibreisundergoingadramaticrevival,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.LinktoArticleReadAbstractDebatecontinuesonthedefinitionofdietaryfiber(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.LinktoArticleReadAbstractInterestindietaryfibreisundergoingadramaticrevivalthanksinparttotheintroductionofnewcarbohydratesasdietaryfibrecomponents.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.LinktoArticleReadAbstractDietaryfiberismainlycomposedofplantcellwallpolysaccharidessuchascellulose,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.LinktoArticleReadAbstractAstudywasmadeoftheeffectoftheactivityandpurityofenzymesintheassayoftotaldietaryfiber(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.LinktoArticleReadAbstractEnzymeactivityandpurityofthesetopics,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.LinktoArticleReadAbstractThe"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.LinktoArticleReadAbstractWiththerecognitionthatresistantstarch(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.LinktoArticleReadAbstractAmethodisdescribedforthemeasurementofdietaryfibre,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.LinktoArticleReadAbstractAnintegratedtotaldietaryfibre(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.LinktoArticleReadAbstractAmethodforthedeterminationoftotaldietaryfiber(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.LinktoArticleReadAbstractAmethodforthedeterminationofinsoluble(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.LinktoArticleReadAbstractTheCodexCommitteeonMethodsofAnalysisandSamplingrecentlyrecommended14methodsformeasurementofdietaryfiber,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.LinktoArticleReadAbstractAOACOfficialMethods2009.01and2011.25havebeenmodifiedtoallowremovalofresistantmaltodextrinsproducedonhydrolysisofvariousstarchesbythecombinationofpancreaticα-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.LinktoArticleReadAbstractThereliabledeterminationofsoluble,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.LinktoArticleReadAbstractDietaryfiber(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.ValidationofMethodsAACCMethod32-45.01AACCMethod32-50.01AOACMethod2009.01AOACMethod2011.25CODEXMethodTypeIForthedeterminationofTotalDietaryFiberincerealproducts,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   andsubtractedKitsize:                         100assaysMethod:                           Hydrolysis/removalofnon-dietary                                         fibrecomponentsTotalassaytime:              ~3hrwork(over2days)Detectionlimit:                 0.5-100%ofsampleweightApplicationexamples:Foodingredients,foodproductsandothermaterialsMethodrecognition:   AOAC(Methods2009.01and2011.25),AACC(Method32-45.01and32-50.01)andCODEX(TypeImethod)AdvantagesTheonlymethodthatisconsistentwiththeCODEXAlimentariusdefinitionofdietaryfibre Highpurity/standardisedenzymesemployed Allreagentsstablefor>2years Mega-Calc™softwaretoolisavailablefromourwebsiteforhassle-freerawdataprocessing Verycompetitiveprice(costpertest) 

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