三聚氰胺、沙丁胺醇及苯乙醇胺a的酶联免疫吸附分析法(elisa)的分析和应用-analysis and application of enzyme linked immunosorbent assay ( elisa ) for melamine, salbutamol and phenylethanolamine a.docx

Studyandapplicationofenzyme-linkedimmunosorbentassays(ELISAs)forthedetectionofmelamine,salbutamolandphenylethanolamineAAbstractInChina,foodsafetyincidents(e.g.melamine,β-agonist)occurredfrequently,makingthefoodsafetybecomeahotissueforpublicconcern.Therefore,itisanurgentdemandtodeveloprapiddetection/screeningmethodsformelamineandβ-agonists.Inthisstudy,highlysensitiveandspeci?cindirectcompetitiveenzyme-linkedimmunosorbentassays(ELISAs)forthedetectionofmelamine,salbutamol,phenylethanolamineAinfoodandfeedsamplesweredeveloped.Melamineisakindofimportantorganicnitrogenousheterocyclictriazinechemicalrawmaterial.Becausemelaminecontainslargeamountsofnitrogenelementinthemolecule,someunscrupulouspersonsaddmelamineinfeed,milkpowderandothervegetableoranimalproteinfoodstoimprovetheirproteincontentoftheproductandtogetmorebenefit.Melamineisoflowtoxicity,butifusingwithcyanuricacid,itwillforminsolublemelaminecyanurate,resultinginseriouskidneystones,leadingtoadversehealtheffectonanimalsandhumansbeing.Inthisstudy,todevelophighsensitivityandspecificityoftheELISAforthedetectionofmelamineinmilk,milkpowderandfeedsamples,rationalhaptenmodificationandheterogeneousantibody/coatingantigencombinationswereadopted.Threemelaminederivativeswithdifferentlengthofcarboxylicspacerattheendweresynthesizedandlinkedtocarrierproteinsfortheproductionofimmunogensandcoatingantigens.Monoclonalantibodyagainstmelaminewasproducedbyhybridomatechnology.Underoptimalexperimentalconditions,thestandardcurvesoftheELISAsformelaminewereconstructedinrangeof0.1-100ng/mL.Thesensitivitywas10-300timesenhancedcomparedtothoseinthepublishedliteratures.Thecross-reactivityvaluesoftheELISAsalsodemonstratedtheassaysexhibitedhighspecificity.FivesampleswerespikedwithmelamineatdifferentconcentrationsanddetectedbytheELISA.Therecoveryratesof72.8–133.0%andintra-assaycoef?cientsofvariationof0.8–18.9%(n=3)wereobtained.TheELISAfor英文摘要三聚氰胺、沙丁胺醇及苯乙醇胺A的酶联免疫吸附分析法(ELISA)的研究和应用milksamplewascon?rmedbyhigh-performanceliquidchromatography(HPLC)withahigh