DietaryFiberAssayKit（食物纤维测定***盒）Grade:forFoodAnalysisStoragecondition:Keepat2-10degreesC.Manufacturer:WakoPureChemicalIndustries,Ltd.Cat.No.Pkg.Size291-59701100TestsSummaryThisproductisforresearchuseonly.Donotadministerittohuman.AsimpleandmodifiedversionofProsky'smethod.ComparedtoconventionalProsky'smethod,itassureshighmeasurementaccuracy.Furthermore,enzymetreatmenttimeisshortened.KitContentsThermostablealpha-AmylaseSolution1vialx20mLProteaseSolution1vialx20mLAmyloglucosidaseSolution1vialx20mLDiatomaceousEarth1vialx100gMeasurementOutlineSampledissolvedinMES/TrisbufferDigestionwiththermostablealpha-amylaseDigestionwithproteaseandamyloglucosidasefor30minutesatpH6.3and60degreesCEthanolPrecipitationLe***e1hourFiltrationWashwithethanolandacetoneDryovernightKjeldahlProteinDeterminationandAshDeterminationTotalDietaryFiberDeterminationPrinciple1、DeterminationoftotalDietaryFiberContentsDigestiblecomponentsinsamples,suchasstarchandprotein,arehydrolyzedwithalpha-amylase,protease,andamyloglucosidase,precipitateundecomposedresidualmacromoleculebyaddingquadruplevolume95%Ethanol,andthentrappedwithaglassfiltercruciblecontainingdiatomaceousearth.TotalDietaryFiberContentsaredeterminedbydeductingweightofproteinandashcontentfromthedryweightoftrappedresidueonaglassfiltercrucible.2、FractionalAnalysisofSolubleDietaryFiberContentsandInsolubleDietaryFiberContentsDigestiblecomponentsinsamples,suchasstarchandprotein,arehydrolyzedwithalpha-amylase,protease,andamyloglucosidase,thenfiltratedwithaglassfiltercruciblecontainingdiatomaceousearth.InsolubleDietaryFiberContentsaredeterminedbydeductingweightofproteinandashcontentfromthedryweightoftrappedinsolubleresidueonaglassfiltercrucible.Meanwhile,addaquadruplevolume95%Ethanolintothefiltrate,precipitateundecomposedresidualsolublemacromolecule,andtrapwithaglassfiltercruciblecontainingdiatomaceousearth.SolubleDietaryFiberContentsaredeterminedbydeductingweightofproteinandashcontentfromthedryweightofthetrappedprecipitatedresidueonaglassfiltercrucible.TotalDietaryFiberContentsaredefinedasthesumofInsolubleDietaryFiberContentsandSolubleDietaryFiberContents.3、DeterminationofLow-molecularSolubleDietaryFiberSpecificLow-molecularSolubleDietaryFibers,whicharenotprecipitatedevenbyadditionofaquadruplevolumeof95%Ethanol,aredeterminedbyhigh-performanceliquidchromatography.Afterdigestiblecomponentssuchasstarchandproteininsamplesarehydrolyzedwithalpha-amylase,protease,andamyloglucosidase,theundecomposedresidualmacromoleculeareprecipitatedbyaddingaquadruplevolumeof95%Ethanol.Thenfilteritwithaglassfiltercruciblecontainingdiatomaceousearth.Proteins,organicacidsandinorganicsaltsinthefiltrateareremovedwithion-exchangeresin,andthenthesolutionisappliedtohigh-performanceliquidchromatography.Low-molecularSolubleDietaryFiberContentisdeterminedbytheratioofthepeakareaofthesamplewiththatofglucose(internalstandard)onthechromatogram.Meanwhile,InsolubleandSolubleMacromoleculeDietaryFiberContentisdeterminedbydeductingweightofproteinandashcontentfromthedryweightofthetrappedprecipitatedresidueonaglassfiltercrucible.TotalDietaryFiberContentsisdefinedassumofLow-molecularSolubleDietaryFiberContentsandInsolubleandSolubleMacromolecularDietaryFiberContents.说明书单询。)