3P-37 PDF
LC/MS analysis of low molecular substances having a phosphate group or a sulfo group in biological samples using polymer-based HILIC column
In this study, a new HILIC column (HILIC or RP + anion exchange) Shodex® HILICpak® VT-50, packed with polyvinyl alcohol base material modified with quaternary ammonium functional groups, was used to develop high-sensitive LC/MS method for the quantification of various anionic low molecular substances. Phosphorylated saccharides such as Glucose-6-phosphate are important intermediates involving energy metabolism. In order to simplify conventional analytical method required derivatization or ion pair reagent, a new LC/MS method was studied. We found that VT-50, with aqueous solution including ammonium formate /acetonitrile as the eluent, was useful for this analysis. Peak shapes of phosphorylated saccharides were sharp with effective separation, and the calibration curves were linear. Both detection limits were 0.005 μM. Moreover, it was demonstrated that this method was applicable for various other anionic substances (e.g. cAMP, AMP, Taurine, Taurocholic acid). The new HILIC column was useful for the LC/MS analysis of various anionic substances using more simple conditions than previous methods.