HIGH PERFORMANCE LIQUID CHROMATOGRAPHY OF MONO- AND OLIGOSACCHARIDES DERIVATIZED WITH P-AMINOBENZOIC ETHYL ESTER ON A C-18-BONDED SILICA COLUMN

摘要

This report examines a behavior of mono- and oligosaccharide-ABEE derivatives in RP-HPLC. Sugar-ABEE derivatives are saparated by reverse-phase partition chromatography rather than normal-phase partition chromatography. Reducing sugars are derivatized with p-aminobenzoic ethyl ester(ABEE) for the UV detection at 254 nm. C18-bonded silica column is used for the separation of sugar-ABEE derivatives in an isocratic mode. RP-HPLC is performed by using ternary mixture as a mobile phase and column temperature is maintained at 45 degrees C. Sugar-ABEE derivatives are separated well within a short run time(ca. 25 min) by reverse-phase partition chromatographic mode, and C18-bonded silica column is very stable during the analysis. The (1-->6)-linkage type of dihexose-ABEE derivatives has shorter retention time than (1-->4)-linkage type. beta-Anomeric configuration of [Glc-beta-(1-->4)Glc]-oligomer has shorter retention time than alpha-anomer in RP-HPLC. Column temperature and ionic strength slightly affect the elution behavior of sugar-ABEE derivatives.