Analysis of oligosaccharides by microbore high-performance liquid chromatography

A 1-mm microbore hydrophilic interaction column has been used for the separ ation of 2-aminoacridone (2-AMAC)-derivatized glycan mixtures, released fro m naturally occurring and recombinant proteins, Primary structure identific ation of the 2-AMAC glycan derivatives was carried out by HPLC using fluore scence and mass spectrometric detection. In some cases, enzymatic digestion of the 2-AMAC glycans was applied to confirm glycan structure. This strate gy is considerably more rapid than methods normally used in glycan analysis , which involves manual collection of separated oligosaccharide derivatives and analysis of individual fractions by mass spectrometry.