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.

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.

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.

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.

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.

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.

Amethodforthedeterminationoftotaldietaryfiber(TDF),asdefinedbytheCODEXAlimentarius,wasvalidatedinfoods.BasedupontheprinciplesofAOACOfficialMethods^SM985.29,991.43,2001.03,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(S_r)rangedfrom0.41to1.43,andreproducibilitystandarddeviations(S_R)rangedfrom1.18to5.44.Theseresultsarecomparabletootherofficialdietaryfibermethods,andthemethodisrecommendedforadoptionasOfficialFirstAction.

Amethodforthedeterminationofinsoluble(IDF),soluble(SDF),andtotaldietaryfiber(TDF),asdefinedbytheCODEXAlimentarius,wasvalidatedinfoods.BasedupontheprinciplesofAOACOfficialMethods^SM985.29,991.43,2001.03,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(S_r),forIDFrangedfrom0.13to0.71,andthebetween-laboratoryvariability,reproducibilitySD(s_R),forIDFrangedfrom0.42to2.24.Thewithin-laboratoryvariabilitys_rforSDFrangedfrom0.28to1.03,andthebetween-laboratoryvariabilitysRforSDFrangedfrom0.85to1.66.Thewithin-laboratoryvariabilitysrforTDFrangedfrom0.47to1.41,andthebetween-laboratoryvariabilitys_RforTDFrangedfrom0.95to3.14.Thisiscomparabletootherofficialandapproveddietaryfibermethods,andthemethodisrecommendedforadoptionasOfficialFirstAction.

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.

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.

Thedietaryfibre(DF)contentinwheatgrainbasedfoodproductshavebeenestablishedwithboththeclassicalAOAC985.29dietaryfibreandthenewAOAC2009.01totaldietaryfibreprotocol.Thereisagoodagreementbetweenthehighmolecularweightdietaryfibre(HMWDF)contentsmeasuredwiththeAOAC2009.01methodand(DF)contentmeasuredwiththeclassicalAOAC985.29methodinwheatgrainbasedfoodproducts.WiththeAOAC2009.01methodalsoasignificantamountoflowmolarweightdietaryfibre(LMWDF),rangingfrom1%to3%w/w,wasmeasuredwhichisnotquantifiedwiththeAOAC985.29method.Withsemi-preparativeGPCtheLMWDF(DP≥3)fractionsinthewheatgrainbasedfoodproductswereisolated.ThemonosaccharidecompositionofthedissolvedLMWDFconstituentswasdetermined.Glucosewasbyfarthemostabundantmonosaccharidepresentwitharabinose,galactose,xyloseandmannoseasminorconstituents.ItappearedthattheLMWDFcontainsstillnotfullyconverteddigestiblestarch/malto-oligosaccharidefragmentswithDP≥3,whichareerroneouslyquantifiedasLMWDF.ByintroducinganextraAMGhydrolysisstepintheAOAC2009.01protocolafterevaporationoftheethanolanddissolvingtheresidueindeionisedwater,thesemalto-oligosaccharidesarefullyhydrolysedresultinginthatwayinacorrectandlowerLMWDFcontent.

ApreliminaryinterlaboratorystudywasconductedtoevaluatethevalidityofthemodifiedAOACmethodfordeterminationoftotaldietaryfiberbyTadaandInnami,inwhichthe3-stepenzymaticdigestionprocessinAOACMethod991.43ismodifiedtoa2-stepprocesswithoutpHadjustment.Totaldietaryfibercontentsin8representativefoodstuffsweremeasuredusingboththeoriginalAOACMethod991.43andthemodifiedmethodin6researchfacilitiesinJapan.Repeatabilityrelativestandarddeviations,reproducibilityrelativestandarddeviations,andHorwitzratiovaluesfromthemodifiedmethodwereequivalenttothosefromAOACMethod991.43,exceptinthericesample.However,thisexceptionalcaseshowninthemodifiedmethodwasentirelydissolvedbytheadditionof-amylasestabilizingagents.Themodifiedmethod,whichshortenstheprocessofenzymaticdigestionfrom3to2stepsandinwhichonlyreactiontemperatureisadjustedunderthesamepH,wasfoundnotonlytogiveaccuratevaluescomparabletotheoriginalmethod,butalsotosubstantiallyreducethelaborrequiredbythelaboratorystaffintheprocessofroutineanalysis.Thisstudyrevealedthatthevalidityofthemodifiedmethodwasfurtherensuredbyadding-amylasestabilizingagentstothereactionsystem.

NutritioNISTsrecommendincreasingtheintakeofsolubledietaryfiber(SDF),whichisverylowinmostcereal-basedproducts.ConversionofinsolubleDF(IDF)intoSDFcanbeachievedbychemicaltreatments,butthisaffectsthesensorialpropertiesoftheproducts.Inthisstudy,thepossibilityofgettingasubstantialincreaseofSDFfromcerealproductsusingatailoredpreparationofTrichodermaenzymesisreported.EnzymeswereproducedcultivatingTrichodermausingdurumwheatfiber(DWF)andbarleyspentgrain(BSG)asuniquecarbonsources.ManyTrichodermastrainswerescreened,andthehydrolysisconditionsabletoincreasebyenzymatictreatmenttheamountofSDFinDWFandBSGweredetermined.ResultsdemonstrateinbothproductsthatitispossibletotripletheamountofSDFwithoutamarkeddecreaseoftotalDF.Theenzymatictreatmentalsocausesthereleaseofhydroxycinnamicacids,mainlyferulicacid,thatarelinkedtothepolysaccharideschains.Thisincreasesthefreephenolicconcentration,thewater-solubleantioxidantactivity,and,inturn,thephenolcompoundsbioavailability.

Starchgranulesisolatedfromwaxy,normal,andhigh-amylosehulllessbarleygrainswereexaminedbytransmissionelectronmicroscopywithcytochemicaltechniques.Themicrographsshowedtwodistinctregionsofdifferentsizes:1)denselypackedgranulegrowthrings(whichvariedinsizeandnumberdependingonthegenotype),and2)aloosefilamentousnetworklocatedinthecentralregionofthegranule.Thegranuleringwidthdecreasedwithincreasingamylosecontent.Inallthreegenotypes,thegrowthringsclosertothegranulesurfacewerenarrowerinwidththanthosewithinthegranuleinterior.Thewaxystarchhadwiderintercrystallineamorphousgrowthrings,semicrystallinegrowthrings,andmoreopencrystallinelamellaethannormalandhigh-amylosestarches.Granuleboundproteins(mainlyintegralproteins)werelocatedinthecentralandperipheral(growthring)regionsofthegranule.

Wheatflourwasextrudedatdifferentconditionsofbarreltemperature(120°Cand180°C),watercontent(18%and22%)andscrewspeed(400rpmand800rpm)withanincreasingconcentrationofwheatbranfibers(2.8%,12.6%and24.4%).Inthetestedextrusionconditions,starchcrystalliteswerefullydissociated.Theestimatedstarchsolubilitywasinfluencedbytheprocessconditionsandrangedfrom24.1%to63.1%.Atsameprocessconditions,thestarchsolubilitywasincreasedonlyatthehighestbranlevel.Thebranconcentrationinfluencedtheglasstransitiontemperature,meltingtemperatureandsorptionisothermoftheunprocessedwheatflour.Attheextrusionconditions,itshowedthathigherbranlevelsledtoahigheramountoffreewaterandadecreaseinstarchglasstransitiontemperatureofupto13K.Thedifferencesinstarchtransformation,inducedbytheconcentrationofbran,mightcontributetothemodulationoftheexpansionpropertiesofbran-containingstarchyfoams.

Dietaryfiber(DF)isoneofthemaindietaryfactorscontributingtoconsumers"well-being.Inthisworkthepossibilityofusingtheroastedcoffeesilverskin(CS),abyproductofroastedcoffeebeans,asaDF-richingredienthasbeenevaluated.Theresultsofourinvestigationshowedthatthismaterialhas60%totalDF,witharelevantcomponent(14%)ofsolubleDF.AlthoughasmallamountoffreephenolcompoundsispresentinCS,ithasamarkedantioxidativeactivity,whichcanbeattributedtothehugeamountofMaillardreactionproducts,themelanoidins.StaticbatchculturefermentationexperimentsshowedthatCSinducespreferentialgrowthofbifidobacteriaratherthanclostridiaandBacteroidesspp.CScanbeproposedasanewpotentialfunctionalingredientinconsiderationofthehighcontentofsolubleDF,themarkedantioxidantactivity,andthepotentialprebioticactivity.

Theeffectofprecookingbyextrusionprocessingonthedietaryfiberprofileofwheatfloursubstitutedwith0%,10%,20%,and30%wheatbranwasevaluated.Dependingonthelevelofbran,totaldietaryfiber(TDF)andsolubledietaryfiber(SDF)inuncookedfloursrangedfrom4.2%to17.2%and1.5%to2.4%,respectively.PrecookingbyextrusionsignificantlyincreasedSDFinflours(by22%to73%);althoughinmostcasesitalsoledtoasignificantdecreaseinTDF.Cookiesandtortillasproducedfromuncookedandprecookedflourswith0%and20%substitutedbranwereevaluatedforconsumeracceptabilityusinga9-pointhedonicscale.Withafewexceptions,allcookieshadscoresrangingfrom6to7(“likeslightly”to“likemoderately”)foreachattribute,includingoverallacceptability,appearance,texture,crumbliness,andflavor.Tortillaswereratedforthesameattributesexceptforcrumbliness,whichwasreplacedwithchewiness.Inmostcases,tortillascoresrangedfrom5to7(“neitherlikenordislike”to“likemoderately”).Consumeracceptabilityscoresofcookiesfromuncookedflourdidnotchangesignificantlywithincreaseinbransubstitutionfrom0%to20%.However,consumerscoresfortortillasdiddecreasesignificantlywithincreaseinbranlevel.Extrusionprecookingofthefloursdidnotimprovetheconsumeracceptabilityofcookiesandtortillas;however,itdidimprovetheirdietaryfiberprofilebyincreasingtheSDFsignificantly.

Solubledietaryfibrewithlowviscosityhasthepotentialtodeliveracceptablemouthfeelandtexturewhenincludedinthedietinasignificantamounttoshowhealthbenefits.Solubleflaxseedgum(SFG)wasreportedtohavelowviscosity,thushaspotentialinapplicationsasafibrefortifier.Inthepresentwork,SFGextractedfromflaxseedhullswasfractionatedintoaneutralfractiongum(NFG)andanacidicfractiongum(AFG)usingionexchangechromatography.TheproteincontentinSFG(11.8%)andAFG(8.1%)werecompletelyremovedbyproteasetoobtaintwomoreprotein-freefractions,SFGnPandAFGnP;NFGcontainednoprotein.TheuronicacidcontentinNFG(1.8%)wasmostlyeliminated,whereasinAFCincreasedto38.7%.NFGconsistedofhighmolecularweight(MW)(1470kDa)arabinoxylansandexhibitedpseudoplasticflowbehaviour;whereasAFGwasmainlycomposedofrhamnogalacturonanswithahigherMWfraction(1510kDa)andalowerMWfraction(341kDa)andshowedNewtonianflowbehaviour.Therankingofintrinsicviscosities(mLg^-1)indecreasingorderwas:SFG(446.0)>NFG(377.5)>AFG(332.5).AFGwasexpectedtohavehigherchainflexibilityforitslowervalueofHugginsconstant(0.16)comparedtothatofNFG(0.54)andSFG(0.48).IncomparisonwithSFGnP,AFGnPandNFG,SFGandAFGshowedbettersurfaceactivitiesandemulsifyingstabilitiesduetothepresenceofproteininbothfractions.

OchratoxinAisanimportantmycotoxinthatcanenterthehumanfoodchainincereals,wine,coffee,spices,beer,cocoa,driedfruits,andporkmeats.Coffeeisoneofthemostcommonbeveragesand,consequently,ithasapotentialriskfactorforhumanhealthrelatedtoochratoxinAexposure.Inthisstudy,coffeeandcorrespondingbyproductsfromsevendifferentgeographicregionswereinvestigatedforochratoxinAnaturaloccurrencebyHPLC-FLD,nutritionalcharacterization,andantioxidantactivitiesbyspectrophotometricassay.Theresearchfocusedoncompositionchangesincoffeeduringtheprocessingstep“fromfieldtocup”.CostaRicaandIndiangreencoffeeswerethemostcontaminatedsamples,with13and11µg/kg,respectively,whiletheEthiopiancoffeewastheleastcontaminated,with3.8µg/kgofochratoxinA.ThereductionofochratoxinAcontaminationduringtheroastingstepwascomparableforanysamplesthatwereconsideredundertherecommendedlevelof4µg/kg.Totaldietaryfibersrangedfrom58.7%forVietnamand48.6%forIvoryCoastingreencoffeesandrangedfrom58.6%forCostaRicato61.2%forIndiainroastedcoffee.CoffeesilverskinbyproductobtainedfromIvoryCoastwasthehighest,with69.2and64.2%ofinsolubledietaryfibers,respectively.

Althoughvasculardementiaisthesecondleadingcauseofdementiaandoftenunderdiagnosed,therearenodrugsyetapprovedforthetreatmentofvasculardementia.Inthisstudy,itisdemonstratedthatwaterextractofTriticumaestivumL.(TALE)andsomeofitscomponentshaveprotectiveeffectsagainstvasculardementia-induceddamagebypreservingthemyelinsheathandinhibitingastrocyticactivation.Thememorytestusedavasculardementiamodelutilizingbilateralligationofthecarotidarteriesofrats.TALE,someofitscomponents,suchasstarch,totaldietaryfiber(TDF),arabinoxylan,β-glucan,anddegradedproductsofarabinoxylan,suchasarabinoseandxylose,wereadministeredtotheanimalsfromday8today14,followingthesurgery.Twenty-onedaysafterthesurgery,thewatermazetestwasperformedfor5days,andthetimetakentofindtheplatformduringtrainingtrials(meanescapelatency)wasmeasured.ThemeanescapelatencywasdecreasedconsistentlyintheTALE-,starch-,TDF-,arabinoxylan-,andarabinose-treatedgroups,comparedwiththatinthevasculardementiagroup.Tomeasurebraindamage,Luxolfastbluestainingandimmunohistochemistryofmyelinbasicprotein(MBP)wereperformedtoobservemyelinsheathinthewhitematter,andimmunohistochemistryofglialfibrillaryacidicprotein(GFAP)wasperformedtoobservetheastrocyticreaction.VasculardementiareducedtheMBPlevelandincreasedtheGFAPlevel.ArabinoseeffectivelyinhibitedtheMBPandGFAPchange,whereasarabinoxylaninhibitedtheGFAPchangeonly.TheseresultssuggestthatTALEandsomeofitscomponentscanbeusedasamedicinalmaterialforthedevelopmentofneuroprotectiveagentsagainstvasculardementia.

Thedistributionofsubstituentsalongthepolymerchainincationicpotatoamylopectinstarch,modifiedinsolution,granularslurry,ordrystate,wasinvestigated.Thestarchderivativesweresuccessivelyhydrolyzedbydifferentenzymes,followedbycharacterizationofthehydrolysisproductsobtainedbymeansofelectrospraymassspectrometry(ESI-MS)andmatrix-assistedlaserdesorption/ionizationmassspectrometry(MALDI-MS).ESI-MSandMALDI-MSwereprovedtobeappropriatetechniquesforidentificationofthesubstitutedhydrolysisproducts,forwhichtherearenostandardcompoundsavailable.Nohighlysubstitutedoligomerswerefoundinthehydrolysates,whichwastakenasanindicationofamoreorlesshomogeneousdistributionofcationicgroupsintheamylopectinmolecules.FurThermore,fromtheresultsobtaineditwassuggestedthattheenzymescleaveglucosidiclinkagesonlybetweenunsubstitutedglucoseunitsand,preferentially,linkagesinsequencescontainingmorethantwoadjacentunsubstitutedunits.Thedeterminationoftheamountofunsubstitutedglucoseproducedfromeverysuccessivehydrolysissteprevealedslightdifferencesbetweenthedifferentstarchsampleswithrespecttothehomogeneityofthesubstitutionpattern.Amongthethreesamplesunderinvestigation,starchcationizedinsolutionwasfoundtohavethemostanddry-cationizedstarchtheleasthomogeneousdistributionofsubstituents.

AnHPLCmethodisdescribedusingnormalphaseconditionswithanunbondedsilicacolumntodetermineconcentrationsofsupplementaryvitaminAandvitaminEestersandβ-caroteneininfantformulae.Themethodutilisesselectivedual-channelfluoresenceforvitaminsAandEandvisibleabsorbanceforβ-carotene.Anattributeofthemethodistheuseofretinolpropionate,α-tocopherylpropionateandall-E-β-apo-8′-carotenoicacidethylesterinternalstandardstocompensateforanalyticalvariationsassociatedwiththeselabilevitamins.Extractionisperformedwithoutsaponification,withtheaidofproteasetoremovevitaminencaspsulationandfacilitatevitaminpartitionintohydrocarbonsolvent.Figuresofmeritindicatethemethodissuitableforitsintendedpurposeinthehighlyregulatedinfantformulaenvironment,includingliquidformulations.ThemethodisextendabletowholemilkpowderswheretotalvitaminAcontentdatacanbecalculatedbysummingtheinnatelong-chainvitaminAesterswiththeaddedesters.

Apartfrombeingarichandinexpensiveproteinsource,legumesprovideessentialvitamins,mineralsanddietaryfibre.Consideringthenutritionalbenefits,legumesflourcanpotentiallybeincorporatedinthedevelopmentofnewproducts.Theaimofthisstudywastoinvestigatewhetherfermentationaffectstheproteincontent,invitroproteindigestibility,trypsininhibitoractivityandthefunctionalityofproteinsinfababean,desiandkabulichickpea.Australiangrownchickpeaandfababeanwereselectedandinitiallysoaked,de-hulled,driedandmilledintoflour.Thiswasfermentedwithlyophilisedyoghurtculturesina30°Corbitalshakerfor16 h.Whileproteincontentsinfermenteddesiandkabuliflourwerelowerthantheirrawcounterparts(p > 0.05),itwassignificantlyhigherinfermentedfababean.Asignificantincrease(9.5 %)ininvitroproteindigestibilitywasfoundinfermenteddesi.Trypsininhibitoractivityinfermenteddesi,kabuliandfababeanreducedby2.7,1.1and4.7 %,respectively(p > 0.05).Overall,theinvitroproteindigestibilityinfloursamplesincreased,whilesimultaneouslyreducingthetrypsininhibitoractivity.Thewaterabsorptioncapacityofthefermentedkabulifloursignificantlyincreasedby11.3 %.Allfermentedfloursampleshadsignificantlyhigheroilabsorptioncapacitythantheircorrespondingrawflourthatwaslikelyduetoincreasedinsolublehydrophobicprotein.Although,thefoamingcapacityinallfermentedfloursampleswassignificantlylowerthantheirrespectiverawsamples,onlyfermenteddesiandfababeanflourshowedlowerfoamingstability(p > 0.05).Thepresentstudysuggeststhatfermentedlegumeflourcouldfulfillthedemandforinnovativeproductsofhighernutritionalvalue.