Separation and quantification of short-chain coenzyme A in plant tissues by capillary electrophoresis with laser-induced fluorescence detection

[摘要] CoenzymeA(CoA)isagroupofactivemetaboliccompoundsthatfacilitateover100chemicalreactionsinanimalandplantcells.Itmainlyservesasanacylcarrierinmanymetabolicreactionsandinitiatesthetricarboxylicacidcyclethatproducesmorethan90%oftheenergyrequiredforlifeprocesses.Measurementsofshort-chainandlong-chainCoAcompoundsinavarietyoftissuesbyusinghigh-performanceliquidchromatography(HPLC)andcapillaryelectrophoresis-ultraviolet(CE-UV)detectionhavebeenreported,butthesetechniquesdonotallowonetosimultaneouslydetermineallthepossiblecoexistingCoAsandtheirderivativesinplanttissueswithsufficientsensitivity.Inthispaper,amethodofquantitativedeterminationof5short-chainCoAsinplanttissuesbyusingcapillaryelectrophoresiswithlaser-inducedfluorescencedetection(CE-LIF)wasdeveloped.Underoptimizedderivatizationandelectrophoresisconditions,differentCoAsthatwerederivatizedwithfluorescein-5-isothiocyanate(FITC)wereseparatedandquantifiedatthepmolelevel.Afusedsilicacapillarywitha75μm(i.d.)×57cmwasusedfortheseparationand150mMboratebuffer(pH9.00)wasusedasabackgroundelectrolyte.Theseparationwascarriedoutat25kVandcompletedinlessthan13min.Theeffectsofderivatizationtime,bufferconcentrations,andpHvaluesonderivatizationefficiencywerealsosystematicallyinvestigated.ThisnewlydevelopedCE-LIFmethodcanbeusedtodetectCoAsinbothplantandanimaltissues...

[学科分类] 分析化学