Development of a highly sensitive method for the quantitative analysis of modified nucleosides using UHPLC-UniSpray-MS/MS

There are more than 150 types of naturally occurring modified nucleosides, which believed to be involved in various biological processes. Recently, an ultrahigh performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UHPLC-ESI-MS/MS) technique has been developed measure low levels nucleosides. A comprehensive analysis nucleosides will lead a better understanding intracellular ribonucleic acid modification, but this requires high-sensitivity measurements. In perspective, we established highly sensitive and quantitative method using the newly ion source, UniSpray. spectrometer was used with UniSpray source positive mode. Our UHPLC-UniSpray-MS/MS methodology separated detected four major 42 dG15N5 (internal standard) 15 min. The provided good correlation coefficients (>0.99) for all analyzed wide range linearity 35 46 Additionally, accuracy precision values satisfied criteria <15% higher concentrations <20% lowest We also investigated whether could samples mouse tissues non-small cell lung cancer clinical specimens. were able detect 43 31 different from tissues, respectively. found significant differences N6-methyl-N6-threonylcarbamoyladenosine (m6t6A), 1-methylinosine (m1I), 2'-O-methylcytidine (Cm), 5-carbamoylmethyluridine (ncm5U), 5-methoxycarbonylmethyl-2-thiouridine (mcm5S2U), 5-methoxycarbonylmethyl-2'-O-methyluridine (mcm5Um) between cancerous noncancerous tissues. conclusion, simultaneously quantify can samples.