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

Karlsson G, Winge S, Sandberg H

Octapharma AB, SE-11275 Stockholm, Sweden. goran.karlsson@octapharma.se

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 degrees 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 microg for each monosaccharide, and intermediate precisions were 3-6% RSD (n=6).

Published 3 October 2005 in J Chromatogr A, 1092(2): 246-9.
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