ForusewiththeTotalDietaryFiberAssayKit.Containsbarleyβ-glucan,highamylosemaizestarch,wheatstarch,casein,pectinandlarchgalactan.WheatarABInoxylanisavailableonrequest.
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“wasviewedasmerelythelessdigest
IBLeconstituentoffoodwhichexertsalaxativeactionbyirritatingthegut“thusacquiringthedesignation“roughage”atermwhichwaslaterreplacedby“crudefibre”andultimatelyby“dietaryfibre”Variousdefinitionsofdietaryfibrehaveappearedovertheyears,partlyduethevariousconceptsusedinderivingtheterm(i.e.originofmaterial,resistancetodigestion,fermentationinthecolonetc.),andpartlytothedifficultiesassociatedwithitsmeasurementandlabelling(4).Theprinciplecomponentsofdietaryfibre,astr
ADItionallyunderstood,arenon-starchpolysaccharides,whichinplantfibreareprincipallyhemicellulosesandcelluloses,andthenon-carbohydratephenoliccomponents,cutin,suberinandwaxeswithwhichtheyareassociatedinNature.
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.
Invitrofermentationofspentturmericpowderwithamixedcultureofpigfaecalbacteria.
Han,K.H.,Azuma,S.&Fukushima,M.(2014).Food&Function,10,2446-2452.
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Thefermentationpotentialofspentturmericwasstudiedininvitroswinefaecalbatchcultures.Thespentturmericresidue(theenzyme-resistantfractionfromspentturmeric,EST)wasobtainedthroughtheuseofthedigestiveenzymesamyloglucosidaseandpancreatinandcomparedtocelluloseandhigh-amylosestarch(HAS)ascarbonsources.ESTshowedsignificantincreasesintotalanaerobes,bifidobacteria,lactobacilliandlacticacidbacteriapopulationscomparedtocelluloseat12,24and48h,andthetotalanaerobiclevelintheHASgroupwassignificantlyhigherthaninthecellulosegroupat24and48h.However,asignificantdecreaseinthecoliformpopulationwasonlyfoundintheHASgroupcomparedtothecellulosegroupat48h.Thetotalshort-chainfattyacid(SCFA)concentrationsintheESTandHASgroupsweresignificantlyhigherthanthatinthecellulosegroupat12hand48h.However,therewasnosignificantdifferenceinthetotalSCFAconcentrationbetweentheESTandHASgroupsat12hand48h.AmmoniaandpHlevelsintheESTandHASgroupsweresignificantlylowerthanthoseinthecellulosegroupat24and48h,buttherewasnosignificantdifferencebetweentheESTandHASgroups.Theseresultsindicatethatthefermentationpotentialoftheenzyme-resistantfractionfromspentturmericiscomparabletothatofcommerciallyestablishedresistantstarch.