The combination of normal phase with reversed phase high performance liquid chromatography for the analysis of asparagine-linked neutral oligosaccharides labelled with p-aminobenzoic ethyl ester.

We have developed a novel approach for the analysis of asparagine-linked neutral oligosaccharides derived from glycoproteins. The oligosaccharides are labelled with p-aminobenzoic ethyl ester and the derivatives are separated on two high performance liquid chromatographic columns, one containing amide-silica and the other containing octadecyl-silica. The elution positions of 39 different ABEE-oligosaccharides on the two columns were plotted on a two-dimensional map. Unique non-overlapping positions of these oligosaccharides demonstrate that this technology would be useful for the identification of Asn-linked oligosaccharides at high sensitivity.