Determination of monosaccharides in glycoproteins by reversed-phase high-performance liquid chromatography on 2.1-mm...

The monosaccharide composition of glycoproteins was determined by using a Supelcosil LC-18 150 x 2.1 mm narrowbore column under reversed-phase conditions. Methodology was developed for sample hydrolysis, N-acetylation, benzoylation, and cleanup prior to high-performance liquid chromatography. Monosaccharides were identified and quantified at picomole levels. Various glycoproteins were then analyzed, including ribonucleases A and B, Mucin, and fetuin. In addition, the monosaccharide content of Chinese hamster ovary cell-expressed recombinant human renin was determined.