Combination of two hydrophilic interaction chromatography methods that facilitates identification of 2-aminobenzamide-labeled oligosaccharides.

Hydrophilic interaction chromatography (HILIC) is used to separate 2-aminobenzamide- (2-AB) labeled N-linked oligosaccharides. The glycans of the model protein, bovine fetuin, are identified following comparison of elution patterns of seven 2-AB-labeled glycan standards, of which two are of the high-mannose type and five are of the complex type. The combination of two HILIC methods, using an Amide-80 column, having different resolutions and selectivities, markedly facilitates the identification of the fetuin glycans. These HILIC methods are suitable for obtaining glycan profiles of complex mixtures.