简介：Inthisstudywesystematicallyanalyzedtheelutionconditionoftrypticpeptidesandthecharacteristicsofidentifiedpeptidesinreversephaseliquidchromatographyandelectrospraytandemmassspectrometry(RPLC-MS/MS)analysis.Followingproteindigestionwithtrypsin,thepeptidemixturewasanalyzedbyon-lineRPLC-MS/MS.Bovineserumalbumin(BSA)wasusedtooptimizeacetonitrile(ACN)elutiongradientfortrypticpeptides,andCytochromeCwasusedtoretestthegradientandthesensitivityofLC-MS/MS.Thecharacteristicsofidentifiedpeptideswerealsoanalyzed.Inourexperiments,thesuitableACNgradientis5%to30%fortrypticpeptideelutionandthesensitivityofLC-MS/MSis50fmol.Analysisofthetrypticpeptidesdemonstratedthatlonger(morethan10aminoacids)andmulti-chargestate(+2,+3)peptidesarelikelytobeidentified,andthehydropathicityofthepeptidesmightnotberelatedtowhetheritismorelikelytobeidentifiedornot.Thenumberofidentifiedpeptidesforaproteinmightbeusedtoestimateitsloadingamountunderthesamesamplebackground.Moreover,inthisstudytheidentifiedpeptidespresentthreetypesofredundancy,namelyidentification,charge,andsequenceredundancy,whichmayrepresslowabundanceproteinidentification.