Reversed-phase HPLC of monosaccharides in glycoproteins derivatized with aminopyrazine with fluorescence detection

A precise method for the determination of monosaccharides is described. Monosaccharides were separated as their aminopyrazine derivatives using reversed-phase HPLC and were detected at 410 nm by a fluorescence detector with excitation at 245 nm. The 8 monosaccharides tested were completely separated in about 20 min, and the detection limits were 45-800 fmol. Particularly, strong fluorescence was observed for N-acetylated monosaccharide and sensitive determination could be carried out. The monosaccharide composition of several glycoproteins was also determined by reversed-phase HPLC and anion-exchange HPLC, and the results were consistent with the reported values.