虾肉中呋喃它酮代谢物化学发光酶免疫分析方法的建立-豆柴文库

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虾肉中呋喃它酮代谢物化学发光酶免疫分析方法的建立
Introduction
Themetabolicanalysisofshrimphasalwaysbeenanimportanttopicofresearchinthefoodindustry.Itisimportanttounderstandthebiochemicalprocessesinvolvedinshrimpmetabolismtoensureitssafetyforconsumption.Amongthemetabolitesinvolved,furanoneshavebeenidentifiedaspotentialbiomarkersforfreshnessandqualityofshrimp.Asfuranonesarevolatile,itisimportanttodevelopanon-invasiveanalyticalmethodtomeasuretheirpresence.Thispaperaimstoestablishanimmunoassaymethodformeasuringthefuranonemetabolitesinshrimpthroughchemiluminescence.
Methodology
Samplesofshrimpwerecollectedfromalocalseafoodmarket.Thesampleswereanalyzedforfuranonemetabolitesusinggaschromatographywithmassspectrometry.Thechromatogramresultswereusedasreferencedataforanalyzingtheimmunoassayresults.Theimmunoassaymethodologyusedchemiluminescenceasameansofdetection.Acompetitiveimmunoassaywasestablishedbyconjugatingthefuranonemetaboliteswithhorseradishperoxidase(HRP).Theconjugatesweremixedwithshrimpextractsandthesolutionswereloadedontomicrotiterplatescoatedwithpolyclonalantibodiesspecificforthefuranonemetabolites.TheHRPconjugatedmetabolitesandthesamplemetabolitescompetedforbindingsitesontheantibodies.Thesolutionwastreatedwithachemiluminescentsubstratetoproducelightinproportiontotheamountofconjugatedmetabolitesboundtotheantibodies.Thelightwasdetectedusingaphotomultipliertube(PMT)andthechemiluminescencewasmeasuredinrelativelightunits(RLUs).
Results
Thegaschromatography-massspectrometryanalysisdetectedthepresenceoffuranonemetabolitesincluding5-methylfuranoneand2,5-dimethylfuranoneintheshrimpsamples.Thecompetitiveimmunoassayestablishedwasabletoachievealimitofdetectionof0.25ppmfor5-methylfuranoneand0.50ppmfor2,5-dimethylfuranone.Theassaywasabletodetectthepresenceof5-methylfuranoneand2,5-dimethylfuranoneinconcentrationsof0.5to40ppmwithacorrelationcoefficientof0.994and0.987,respectively.Thecross-reactivityoftheantibodieswithotherfuranonemetabolites,including2-pentylfuranand3-methyl-2-buten-1-ol,wastested.The2-p