Separation of monosaccharides by hydrophilic interaction chromatography with evaporative light scattering detection

Hydrophilic interaction liquid chromatography (HILIC) was used to separate monosaccharides that are common in N-linked oligosaccharides in glycoproteins and other compounds. A TSKgel Amide-80 column was eluted with 82% acetonitrile, in 5 mM ammonium formate (pH 5.5). Column temperature was 60 °C and evaporative light scattering was used for detection (ELSD). With this method, l-fucose, d-galactose, d-mannose, N-acetyl-d-glucosamine, N-acetylneuraminic acid, and d-glucuronic acid were separated, with detection limits of 0.3–0.5 μg for each monosaccharide, and intermediate precisions were 3–6% RSD (n = 6).