Separation of sialyl-oligosaccharides by high-performance liquid chromatography: application to the analysis of mono-, di-, tri-and tetrasialyl-oligosaccharides obtained by hydrazinolysis of α1-acid glycoprotein

Abstract High-performance liquid chromatography has been applied to the separation of isomers of mono-, di-, tri- and tetrasialylated oligosaccharides derived from α 1 -acid glycoprotein by hydrazinolysis. The separation of the sialyl-oligosaccharides on the basis of their negative charges was carried out with quaternary amine-bonded silica. Within each class, the anionic oligosaccharides were fractionated on the basis of their net carbohydrate content on alkylamine-modified silica using a mobile phase consisting of a mixture of acetonitrile and potassium dihydrogen phosphate with 0.01% of 1,4-diaminobutane or 0.01% of tetraethylpentamine.