参考文稿教程current trend technologies detecting food adulteration

Currenttrendinindetectingfood

Asweallknowthesituationoftheillegaladultioninfoodis ingmoreandmorehascausedlargedamagetonotonlylegalfoodprocessors,butalsotheconsumerswhosehealthwillbethreatened.Inthiscase,thetechniquesappliedinthefoodauthenticityshouldbemoresophisticatedsoastomakedefinitiveverificationofthefoodadul ysisCurrent chainreaction(PCR)istheconventionaltechniqueandiscommonlyusedinmanyfieldsofConventionalPCRPolymerasechainreaction(PCR)isthemostcommonlyusedtechniqueinmanyfieldsPCRisatechniquethatamplifiesthespecificareasofDNAwhichweliketoInordertoimprovetheefficiencyofauthentication,ltiplexPCRsdevelopedrecentlyandhasbeenusedinmanyareas,suchasthedetectionofAsisaccurateandstable,PCRmethodisindispensableforthelabelingoffoodproductsandhasbeenappliedinfoodidentification.IthasbeenshownthatthemultiplexPCRcouldbeusedinthesimultaneousidentificationofmultiplemeatspecies.ItisanticipatedthatbyapplyingtheproposedmultiplexPCRmethodforidentifyingchickenpasteinsausagesandmeatproducts,thiskindoffraudcanbeinspectedandeffectivelyprevented[1].AndforthepurposeofimprovingtheconventionalPCR,techniquesbasedonPCRcomeupforfoodauthentication.NovelPCRPCR-RestrictionFragmentLengthPolymorphism(RFLP)PCR‐RFLPtechnique(restrictionfragmentlengthpolymorphism)differsfromysisofPCRproducts,whichrevealsthevarietyofspeciesbythe eofdifferent‐‐247+‐‐‐‐‐323+‐‐329+‐ ‐247+‐‐‐‐‐323+‐‐329+‐Fromtable1,wecangettheinformationthatAluIenzymegeneratedfragmentsof359and97bpincattle,246and210bpfragmentsinbothsheepandgoatandnofragmentsinbuffalo,asperexpectation.HhaI,ApoIandBspTIwerespecificonlytobuffalo,sheepandgoatgenerating247and209bp,329and127bpand323and133bpfragments,respectively.Theresultswererepresentativeof10separateexperimentswithdifferentsamples.Fromthiscase,wecouldconcludethatthesefourcommercialspeciescanbeidentifiedanddifferentiatedbyAnotherAnotherusingPCR‐RFLPistheapplicationforpufferfish‐basedcommercialLnewset1:5‘‐CTTCCTACCCCCTCAAACATTTCHGCMTGRTGAAA‐3’withaproductof376‐bpfragmentofthecytochromebgeneafterPCRFiverestrictionendonucleasesincludingBsaJI,AciI,HinfI,TaqIandSapIwereselectedforspecies‐specificrestrictionpatternsysisoftheamplifiedPCRproducts.TherestrictionfragmentswerevisualisedandphotographedbyUVtransilluminationandtheirsizeswereestimatedbycomparingwithacommercial100‐bpladder[3].Astheresult,sequencesysisofdistinctspecieslikeLagocephalusgloveri,Lagocephaluswheeleri,LagocephaluslunarisandTakifuguoblonguscanbedetected.ItapprovedthattheThismethodhasbeenprovedtobeusedincanidentificationofythepufferspecieswithaccuracyofItprovesthatthemethodusedinthisstudyStudieshasshownthatPCR‐RFLPwas[JH17Movetothebackofsection.这个表格可以横着弄，用一]:[JH19Listthepapersappliedthismethod[JH17Movetothebackofsection.这个表格可以横着弄，用一]:[JH19Listthepapersappliedthismethodintofood ]:1.Itisvulnerable sheep,2.Itishardto therestrictionsite thedetermination productsasthe 3.Itisdifficultdevisethepattern negativewilloccur pletedigestion Itislessstableand designationisnotstandardizedbothin numberandalsotheconditions,damagingthecomparabilityofdatafromdifferentFalsepositiveandnegativeresultsareinevitableduringtheprocesswhichaffectsthereliabilityItisasensitive methodofgeneticitcanmarkmorespotsand informationthanisozymeandRFLPloquat;potato;vegetables;fish;Itrequires lessgenomicBarleyequipmentsand HighDNApurityandenzyme 2.itcandetect blottingand knowledgeof1.ithashigher detection,andlesscarry‐overMeatproducts;wheatspiecesPCR-RAPDRAPD(randomlyamplifiedpolymorphicDNA)isasensitiveandefficientmethodofgeneticmarker;itcanmarkmorespotsandrevealmoreinformationthanisozymeandRFLPfingerprinting.Withoutcomplicatedstepsofhybridization,isotopelabelingorcloningpreparation,RAPDisusuallydominantinheritanceandcouldbecarriedoutonthewholegenome,thussharinglesslimitationonthepartandsizeofthesamplescollectedfrommaterials.(JiaZhang2011)[13].ItisalsotruethatDNA‐basedRAPDmarkershavebeenZ.Q.Mei(2014)[14]developedanimprovedRAPDmethodinordertodeterminethegeographicaloriginsofD.Longansamplesfrom5provincesinsouthern.Theselectiverandomprimerscouldproducedistinguishablebandsthatdefiniyseparateddistinctspecies.TheabundantDNApolymorphismsshowedinthisstudyimpliesthatlonganaccessionscanbedistinguishedbyusingtheimprovedRAPDmethodwhichprovidesproofforfruitResearchersalsoyzedgeneticdiversityofpotatobyusingRAPD,whofoundthatatotalof128uniqueRAPDfragmentsamplifiedfromPCRanddecamerprimerswereobservedamongthe28potatogenotypes.Similaritymeasuresandprincipalcoordinateysisgenerallyreflectedtheexpectedtrendsinrelationshipsofthefullandhalf‐sibpotatogenotypes(Demeke,T.Notonlyappliedinfruitandvegetableidentification,RAPDisalsoapplicableforfishanddiaryproduct.InastudyconductedbyZhouandLiin2006,theydetectedthedifferencesbetweenPelteobagruseupogonandPelteobagrusbyRAPDandclusterysiswhichproofedtheefficiencyofthemethodforthetwofishspeciesidentification[19].Xu&Chen(2008)providedanewmethodtoidentifythemilkoriginratherthantraditionalmethods.Despiteitshighefficiencyandsensitivityindetection,convenienceinsamplescollection,RAPDmethodalsohassomeshortages(table2)PCR-AFLPAFLP(amplifiedfragmentlengthpolymorphism)isdevelopedunderthecombinationofbothRFLPandRAPD,soitdoesnotonlysharethereliabilityandrepeatabilityofRFLP,butalsoinheritthesafety,efficiencyandconveniencefromRAPD[20].AFLPismainlyusedtoidentifythespeciesofcloserelatednesswhichareoftenusedinfoodfraud.AndthedendrogramcreatedbyclusterysisonthebasisofUPGMA(thesametheoryasmentionedinRAPD)canalsorevealtherelationshipofdifferentspeciesforfoodsubstitute.A.AssefaandM.T.Labuschagne[21]hadsuccessfullyappliedAFLPmethodtoShewainEthiopia,andfoundthatthefivefarmers’barleycultivarsaremorphologicallysimilarinthegrainmarket.Inrecentyears,AFLPremainsapopulartechniqueforbarleyauthenticity,andjustin2014,VdovychenkoZhVappliedthismethodtotestfor19varietiesthatindicatedahighsensitivityofthedevelopedsystem.Inaddition,thedendrogrammadeintheirexperimentrevealedaseparatedoriginofvarietiesofmaltingandfeeddirections[22].Atthesametime,farinItaly,researchersassessedthegeneticdiversityandrelationshipsinasampleof54maizelandraces(ZeamaysL.)bymorphologicaltraitsandAFLPprofiling.Intheirstudy,AFLPmarkersproducedahighfrequencyofpolymorphicbandsandwereabletoclearlyfingerprinteachofthelandracesconsidered.Thedatarevealedalargegeneticheterogeneityforbothmorphologicalandmoleculartraitsintheaccessions(HartingsH2008)[23].TherearestillsomeshortagesofAFLPSimpletativetechnologyforfoodauthenticityhasgrownininterestinrecentyearssinceiteliminatetheprocessofgeleletrophoresisandtheadverseeffectofusingtoxicdye,EB(ethidiumbromide).Morerecentlyreportshavefocusedontheuseofefficientreal‐timePCR(RT‐PCR)methodbyusingspecificprimers,namely,TaqMantechnology,orSYBRGreenDyetechnologywithouttheneedforindividualprobedesign.ficationofDNAsequence.Real‐timePCRmethodhaspreviouslybeenreportedbysomeresearchersfortheidentificationofanimalderivedmaterialinmeatmixturesbyamplifyingmtDNAgenesas12SrRNA(Rodriguezetal.,2005),cytochromebgene(Chisholmetal.,2005;Dooleyetal.,2004;Hirdetal.,2004),and16SrRNA(Sawyeretal.,2003).However,noreporthasbeenpublishedforthereal‐timePCRtechniquetargetingthemitochondrialND2andND5genesforthepurposeofspeciesidentification[27].Z.Kesmen(2009)andhiscolleaguesstudiedtherealtimePCRassayfortheidentificationofrawandcookedmeatproductsandsuggestedthattheTaqManprobeusedinthisresearchmightbearapidandsensitivemethodfortheroutinemeatspeciesidentificationsstudiesinraworcookedmeatproducts,thatallowedthedetectionofaslittleas0.0001ngtemteDNAfrompuremeatforeachspeciesinvestigatedandexperimentalmeatNotonlyusedinthedetectionofmeatproducts,realtimePCRhasalsobeenappliedfortheidentificationofwheatspecies.Commonwheatishexaploidandhasthreegenomes(A,BandD)whereasT.DurumistetraploidandlackstheDgenome(Bryan,G.J1998)[28].Asequence(PSR128)hasbeenidentifiedthatshowsasignificantlevelofsequencepolymorphismbetweenthethreegenomesandlittlepolymorphismwithinthegenomes.Inparticular,thereare54bpwithinanintronthatispresentintheDgenomebutnottheAandBgenomes.ThusthelengthofthisintroncanbeusedfordetectionoftheDgenomeandaconservedregionofthePSR128sequencecanbeusedasaninternalstandard.Forfication,asetofreferencepastaswaspreparedwithhardflouradultedtodifferentdegreeswithsoftflour[29].SpectroscopeSpectroscopeisregardedthatInfraredspectroscopy(IR),Ramanspectroscopy,Nuclearmagneticresonancespectroscopy(NMR),Ultravioletandvisiblespectroscopy(UV)arefrequently‐usedSpectroscopictechniquesinyses.Fortheirstrengthsindetectingthestructureandcompositionoforganics,themethodsmentionedabovearebeinginalargeapplicationinfoodadultion,andtheyhavealsodevelopedindetectingadultioninfood.Andthispartshowshowthesetechniquesapplyinadultiondetecting.IR,duetoitsrapidity,simplicityandlowfinancialcost,hasbeenanemergingyticalmethodfordeterminingtheauthenticityoffoodsamples.TheforemostapplicationofIRisbeenappliedtotativeysiscoupledwiththeintensivelystudiedchemometrics.Mid‐infrared(MIR)spectroscopyonwhichthefundamentalabsorptionofmostorganismisstrongestandthepeaksoffunctiongroupsarenarrowaswellasintenseisusedtodeterminedFoodsthathaverecentlybeendetectedforauthenticityusingFTIRcombinationwithchemometricsincludelotusrootpowderinterfusedwithpotatostarch(JiaLiuetal.,2013),liquidmilk,infantformulaandmilkpowdermixedwithmelamine(NitishRaietal.,2014),virgincoconutoilsneakedincornandsunfloweroils(AbdulRohmanetal.,2011),butteraddingmargarine(Kocaetal.2010),nhoneyadultedwithjaggery(Mishra,S.etal.2010),meatadultion(Rohman,A.2011).NIRinconjunctionwithPartialLeastSquares(PLS)hasbeenusedinthedeterminationofpolymerisedtriacylglyceride(PTG)invegetableoils,providingapredictionerrorof2.28%(w/w),anditwasconsideredasasimple,fast,environmentallymethod(JuliaKuligowski,2012).ErikaMellado‐MojicaandMercedesG.LópezfirstusedNIRandNIRspectroscopycoupledwithPrincipalcomponentsysis(PCA)todistinguishagavesyrupfromthoseadultionsmixedwithstarch,molasses,glucose,dextrin,fructoseorothersugars.IthasbeenfoundthatNIRspectraovertheregionfrom8000to4000cm‐1wasnotabletodistinguishagavesyrupsamongnaturalsyrupswhilethereissignificantdifferencesamongthenaturalsweetenersintheMIRspectrarangefrom1500to900cm‐1(ErikaMellado‐Mojica&MercedesG.López,2015).Withregardstodetectingmelamineadultionofmilkpowder,studiesshowedthatMediumandnearinfraredspectroscopycombinedwithchemometricsreachedtheparallelefficiency(0.76±0.11ppm)(ElisângelaDomingoetal.,2014;RomanM.Balabin&SergeyV.Smirnov,2011).ficationofdiesel/biodieselblendsbyvegetaloilusingNIRhasbeenfoundtobebetterthanMIRintermsofpredictionerrorswhencomparingtheinfluenceofthepretreatmentandselectedvariablesrange(V.Gaydou,J.Kister,N.Dupuy,2011).RamanbetweenphotonandmoleculeaftersubstancesabsorbingirradiationandcreatinginelasticItisreportedthatRamanspectroscopyissuitabletodetectaqueoussolutionduetothenegligibleimpactofwateronRamanspectrum,andwithotheradvantagessuchaseasilysampleHatlen,2004).Butter,withcompositionofmilk,proteinandwater,mixedwithmargarinewasyzedbydemonstratedthatintenseandparticularabsorptionofbutterandmargarinerangedfrom400cm‐1to1500cm‐1(ReyhanSelinUysal,2012).Moreover,Ramanspectroscopyhasalsobeenappliedindetectingothermeatssuchasfish(AnaM.Herrero,2008;Marquardt,B.J.2004),pork(Lyndgaard,L.B.2012)andchicken(EllisDI,2005).Itwasfoundthatinthesurfaceofmetallicsubstratessuchasgold,silverandcopper,theabsorptionsignalsofRamanscatteringhasenhanced104‐106timesfromtheordinaryscattering(SERS).Inrecentyears,withhighsensitivityandselectivity,SERSasanoveltechniquehasincreasinglybeenusedtoysiswidelyincludinghoneyadultedwithsyrups(ShuifangLietal.2012),ractopamine(Zhai,F.,2011),beef(Il‐HoonCho,2014),andchilipowdersneakedinsudanIdye(SimonA.Haughey,2014).Nuclearmagneticresonance(NMR)NMRspectroscopyreferstoaphenomenonofresonancetransitionamongenergyleverscertainfrequencyelectromagneticwavesinanexternalmagneticfield.penetrabilityandprecision,amajoritynumberofresearcheshavebeencarriedoutusingNMRspectroscopyforauthenticationoffoodproducts.Thoseproductscontainhoneywithlow‐costsugars(K. b,2013;).1HMNRhasbeenwidelyemployedinyzingorganicstructures.1HMNRhasonceusedtodetectredwinessneakedinanthocyanin(E.Ferrari,2011).Itwasprovedtobeatechniquewithspeedandeasyacquiringdata.T.Parkeretal.presentedthefirstresultfromanewmachine60MHz1HMNRbench‐topspectrometer,detectingoliveoiladultedwithhazelnutoil(Parker,T.,2014).ThespectrometerownsagreatstrengthofspecificchemicalNMRspectrumthatitprovidedarapidscreeningtoolfordetectinghazelnutoiladultionofoliveoilat11.2%w/w.Itisbecausetheskeletonofalmostorganicsiscomposedofcarbonatomthatfurtherstudiesonthesignalsofcarbonatomusing13CNMRisextraordinarilysignificanttoorganicssignalandwide‐areachemicalshift,andsincethedevelopmentofpulseFouriertransformtechnique,thosedisadvantagesmentionedabovecouldbe eandthesensitivityof13CNMRisimprovedsignificantly.UV–visUV–visspectroscopyisbasedonthetheorythatwhenasubstanceisirradiatedwithabeam(generallyintheultravioletfrom200‐400nmorvisiblelightfrom400‐800nm),moleculewithinthesubstancewilltransittohigherenergylevelinoneofanumberrecognizedwayssuchasn→σ*,n→π*,π→π*thatcangeneratespectrumlines,additionally,thesamplecanbetativebasedontheLambert‐Beerlaw.Duetothecharactersmentionedabove,UV–vis,amethodwithlow‐cost,convenienttooperate,andrapid,hasbeenmainlyappliedtoyzeorganicswithconjugatestructure.TheapplicationsofUV–vishaveincludededibleoiladultedwithwasteoil(JoséS.Torrecilla，2013,Lichen,2014),faketequilas(UlisesContreras,2010).Melamineinthemilkwasis2.32μM(3σ)(Hong,2012).Inarecentstudy,however,employingUV–vis,NIR,andMIRtodetectmincedbeefmixedwithturkeymeathaspointedoutthattheUV–visresultswerelesssatisfactorycomparedwithNIRandMIR(CristinaAlamprese,2013).IthasbeenpointedoutthatthelimitationofUV‐visisthelesssignalabsorptionOnthepurposeof ingtheshrotagementionedabove,UV‐visspectroscopyincombinationwithHighPerformanceLiquidChromatography(HPLC)hasbeenemployedinfoodFejösetalusedHPLC‐UVtodetectPDE‐5inhibitorcontentofillegalerectiledysfunctionproducts(Fejösetal.,2014).Intheresearch,14designerPDE‐5inhibitorswereseparatedbyHPLC–UV,andthemethodwasprovedtobeaalternativetoLC‐MSduetoitsspecificity,ChromatographicChromatographictechniquesincludingliquidchromatography(LC)andgaschromatography(GC)aredefinitiveandwidelyusedinalotoffoodauthenticity.BasedonthetraditionalLC,HighPerformanceLiquidChromatography(HPLC)conquerstheshortagesliketimeconsumingandlaborconsumingoftraditionalLC.AndGCiscommonlyappliedinthefoodsamplesthatarevolatileorsemi‐volatile.AndalotofresearcheshaveprovedthatchromatographictechniquesbasedonLCandGCaremakingprogressindifferenttypesofadultiondetectioninrecentyears,whiletheyalsohavesomedisadvantagestechnically.TheapplicationofHPLChasbeendevelopedinrecentyearsasmanyresearchesreported.ItseemsthatHPLCysasignificantroleinfooddetection.Foods,unlikedrugs,arecomplexmatrices,comprisingamultitudeofcompounds,whicharepronetovariationduetoenvironmentalfactorsandmanufacturingconditions,thusmakingysisachallengingtask(DirkSteinmann,2010;VitaDiStefano,GiuseppeAvellone,2012).Amongthehighofdetection,HPLC–MSistheforemostoneofthisyticalchallenge;beingapowerfultoolforreliableauthenticationofcomplexmixtures(VitaDiStefano,GiuseppeAvellone,2012).AstudydevelopedanHPLC–MS/MSmethodwithone‐stepextractionprocedureforthesimultaneousdeterminationofeightillegalsyntheticdyes(Sudan(I–IV),ParaRed,RhodamineB,ChrysoidinandAuramineO)inchiliproducts,whichhasagoodrepeatabilityandhighaccuracywithlowdetectionlimitsandficationlimits(JuanLi,2013).Subsequenthighpressureliquidchromatographywithultraviolet/visible(HPLC‐UV)aminoacidprofilingfurtherconfirmedtheadultionofthosematerialscontaminatedwithmelamineandmelamine‐relatedcompounds(LawrenceR.Levinson,2011).ThestudyprovesthatHPLC‐UVisindispensableasastand‐alone1stlevelofscreeningtoassesstheintegrityofaVPPoranynutritiveprotein‐basedsample(LawrenceR.Levinson,2011).Futureresearchanddevelopmentisrequiredtobringtheassociatedinstrumentationcostsdowntoalevelwheretheycanbeadoptedonawidespreadbasis(LawrenceR.Levinson,2011).Inordertotesthealthfoodsforillegallyaddeddiureticsforweightloss,H.Wooandhiscolleaguesdevelopedsimple,rapid,selective,andsensitivemethodsusingHPLCandLC‐MS/MSforthesimultaneousysisof17diureticsindietarysupplements(H.Woo,2012).GC–MSwasthefirsthyphenatedtechniqueintroducedin1960s.VolatilityandthermalstabilityarethetwomaincriteriaforanycompoundtobesuccessfullyyzedbyGC–MS.AlthoughonlysporadicreportscanbefoundinlitureforitsuseintheysisofPDE‐5inhibitorsdespiteitslowcostcomparedtoLC–MS(DhavalkumarNarendrabhaiPaa.2013;K.Saisho,2001;C.N.Man;2009;I.Papoutsis,2011;S.Strano‐Rossi;2010).Nevertheless,insituationswhereLC–MSysisaloneisnotabletodefinethestructureofanogueoritsfragment,GC–MSysisafterderivatizationand/orchemicalreactionssuchashydrolysiscanbeusedtocharacterizethestructure(DhavalkumarNarendrabhaiPaa;Forexample,acidhydrolysisfollowedbyGC–MSysiswasemployedtoelucidatestructuresofpiperidinafil(DhavalkumarNarendrabhaiPaa,2013;J.C.Reepmeyer;2006),noracetilde‐nafil(DhavalkumarNarendrabhaiPaa,2013;J.C.Reepmeyer;2007),aildenafil(DhavalkumarNarendrabhaiPaa,2013;J.C.Reepmeyer;2007),thioaildenafil(DhavalkumarNarendrabhaiPaa,2013;J.C.Reepmeyer;2009)anddesulfovardenafil(DhavalkumarNarendrabhaiPaa,2013;Y.H.Lam;2008).Andthereareotherapplicationsinotherfieldsofauthenticationintheseyears.Toinvestigateadultionincommercialchilipowder,thevolatileorganiccompoundsofhealthyandinfectedpowderedchilipepperwerecharacterizedusingasolvent‐solidinjector(SFSI)coupledwithgaschromatography/massspectrometry(GC/MS)(Ah‐YoungKo,2014).Volatilesdifferentsamples(wholeseed,flour,andhusks)wereextractedwithsimultaneousextractionanddistillationbyLikens–NickersonapparatusandyzedbyGC‐MS.Thedetectionofadultionofhighpricedoilsisaparticularconcerninfoodqualityandsafety(LiangxiaoZhang,2014).Inthisstudy,fattyacidprofilesoffiveedibleoilswereestablishedbygaschromatographycoupledwithmassspectrometry(GC/MS)inselectedionmonitoringmode(LiangxiaoZhang,ImmunoassayImmunoassaymethodsareindispensableinfoodchemistryduetotheirofferingasimple,semi‐tativetativeandfastscreeningforroutinedetectionoffoodadultioninfoodproducts.Theenzyme‐linkedimmunosorbentassay(ELISA)issupposedtobethehighlyefficienttechniqueofimmunoassaymethods.AndthedevelopmentofELISAhasalsobeenshowninmanyresearches.Asimmunoassaymethodsareconvenientandefficient,theyhavebeencommonlyusedtheseyears,andwewillfocusonELISAandotherimmunoassaymethodssuchasbiosensorimmunoassaymethodappliedindifferentareasoffoodadultion,andmeanwhile,wewillcomparetheirstrengthandshortages.TheELISAisoneofthemostextensiveimmunoassaytechniquesthathavebeenusedtodetectresiduesinfoods.SomeadvancesinELISAmethodscurrentlymakeitpossibletoscreenetal.,2010;Juxiangetal.,2010;Kim,Perkins,＆Bushway,2008;Petraetal.,2011).Melamineoncewasusedasadditiveinmilkandmilkpowderduetoitsspecialchemicalstructure,andresearchescurrentlyholdtheviewthatthetolerabledailyintakewouldbebetterifunderthelevel0.2mg/kgofbodyweightwhichestablishedbytheWHO(ERICA.E.GARBER,2010;Chen,B,2009;Li,G,2010).ELISAhasbeenregardedasacommonmethodtodetectmelaminethatmanykitshavebeendevelopedforthispurpose.AbraxisMelamineteKitandRomerAgraQuantMelamineSensitiveTestKitusedtobecomparedtogetherinaDespitetheconveniencelikelow‐costandtheabilityofhighoutputthatELISAprovides,itinvolvesmanycomplexproceduresduringcommonysis,suchaswashingandincubation,etc.ToincreasethesensitivityandspecificityofELISAforthedetectionoftargetmoleculesinfoods,acombinationofrationalhaptenmodificationandheterogeneousantibody/coatingantigenhasbeendevelopedinthedetectionofMAinfoodproducts,suchasmilk,milkpowderandfeedsamples,andthereportclaimedthatthismethodhadaexcellentspecificityandAsthetechniquesdevelo,lowerdetectionlimitandhigher‐throughoutscreeningadvancedELISAmethodshavebeenreported.Forinstance,animmunoliposomalnanovesicle(IMLN)basedonimmunomagneticbead(IMB)methodhasachievedasuccessfullyapplicationtothedetectionofgliadininglutenfromwheat.Theimmunomagneticbead(IMB)hasbeenappliedfortheextractionofaimedytefromliquidsamplesasahighperformancematerial.Andtheresultprovedthattheassayhasalowdetectionlimitfor0.6μg/mlofgliadin,althoughitrepresentedslightcross‐reactionswithbarelyandryeconservativeELISA[2].ComparedtotheconservativeELISA,theIMB‐basedELISAismoresensitivewithdetectionlimitofcarcinoembryonicantigeninhumanserumaccordingtotheOtherImmunoassayTherearestillmanyotherimmunoassaymethodsthatmakeupforthedeficiencyofELISA,likethebiosensorimmunoassaymethodandlalflowimmunoassay(LFIA).Inmelamine(MA)detecting,fortheaimofdevelotheeasy,rapid,andcosteffectiveyticaltechnique,thebiosensorimmunoassaymethodhasbeenappliedtodetectMAinmilk(Fodeyetal.,2011),meanwhile,afluorescencepolarizationimmunoassayalsohasbeenusedforthedetectionofMAinmilkpowder(Qiangetal.,2011).Comparedtothesetwoimmunoassaymethods,lalflowimmunoassay(LFIA)ismorerapid,sensitiveandlowcost,ithasbeendevelopedonthebaseofcompetitiveformat,MAandcyromazine(CA)ofwhichMAisthemajormetaboliteinanimalfeedandthecontrolofcropproductioninanimaloriginalfoodsareabletobefoundoutbyusingLFIA(Taoleetal.2012).Meanwhile,aLFIAteststripbasedoncolloidalseleniumhasbeenreportedtodetectMAinmilkproductsandotheranimalfeed,andtheprincipleisthatcolloidalseleniumparticlescanbindtoMAantibodiesandcolloidalseleniumimmunoassayisthebaseofLFIAstrip.Moreover,theMAteststripcanbestoredstabilityfor1yearinadryanddarkenvironment,andithasnotfoundanyfalsepositiveconcequences(ZhizengWang,2014).InordertodeveloptherapidandcommercialdetectionmethodofSudanredⅠinfoodproducts,suchastomatosauceandchillipowder,anotherstripassaythatbasedonthemonoclonalantibody(Mab)labeledwithnanocolloidalgoldcalled8A10hasbeeninvented.Incanbereflectedontheintensityofformedcolorredinthetestline.Itwouldberapidforthe10minysistimeandavisualobservationatalimitationof10ng/g,butthistripisjuststablefor2monthsplusat4℃forstorage(JiaWang,2012).Thecombinationofimmunochromatographicassaywithnanocolloidalgoldhasbeenwidelyusedforthedetectionoflowmolecularweightcompounds(JiaWang,2012;Lisaetal.,2009;Wangetal.,2005;Zhouetal.,2009).Althoughthismethodissemitive,itisverysuitedfortheon‐the‐spottesting(JiaWang,2012).IsotopeysisIsotopeysismethodistheidentificationofisotopicsignature,thedistributionofcertainstableisotopesandchemicalelementswithinchemicalcompounds.Thiscanbeappliedtoafoodwebtomakeitpossibletodrawdirectinferencesregardingdiet,trophiclevel,andInternalstandardstablecarbonisotoperatioysis(ISCIRA)methodhasbeenusedastheAssociationofOfficialyticalChemists’(AOAC)methodforthedetectionofhoneyadultion.Itisappliedbythecomparisonoftheproteinfractionswithdifferentcarbonisotoperatios(URSˇKAKROPFetal.,2010).Andthe13C/12Cisotoperatioysismethodhasbeenusedtodetectsugarcaneorcornsugarsyrupsinhoneyasadultion(MuratTosun,2012;White＆Winters,1989).Thismethodcanbeappliedinthedetectionofadultioninmulberrypekmez,andthestudyhasfoundthatinvertedadultionslikeglucosesyrup(GS)andhighfructosecornsyrup(HFCS)canbedetectedoutbyusingthismethod(MuratTosun,2012).Tomakeimprovement,isotoperatiomassspectrometrycanalsolinkedwithhighperformanceliquidchromatography(HPLC)todetecttheauthenticityoflemonjuiceandhoneyadultion,whichprovidedhighsensitivityandspecificity(FrancoisGuyonetal.,2012;FEIXiaoqing,2011).beenwidelyusedasaclassicseparationmethodtoindentifyproteinsbasedonprotein’smolecularIthasbeenreportedhavinganexcellentseparationresultsofdairyproducts,suchasJovanovic,S.,2007).SDS‐PAGEcanalsobeusedintheysisofhazelnutoil(HAO)asadultioninextravirginoliveoil(EVOO),forthereasonthathazelnutproteinsinsolvent‐extractedHAOcanbedetectedinSDS‐PAGEatthebandsrangingfrom10‐60KDainedEVOOevenatalevelof1%contamination(M.Arlorioetal.,2014).teinsonthebaseofprotein’smolecularweightwhichcanseparatelow‐molecular‐weightproteins,suchasPAPsfrombeef,pork,andturkeyinmeatmixtures.(G.Kreuz,2012;N.Z.Ballin,2010;Vallejo‐Cordoba＆Cora‐Rivas,1998).ComparingwiththePAGE,Capillaryelectrophoresis(CE)providesmanyadvantagesoverPAGE,suchastheshortertimeforysisanddetection,theon‐columnficationandincreasedefficiencyandhigherresolution(ALEJANDROHERNAÄNDEZetal.,2006;Cancalon,P.F.,1995;Manabe,T,1999).Microscopicobservationusuallydependsontheshapeandsizeofsamples,whichissimplebutpowerful.Researchersinusemicroscopet