High Performance Liquid Chromatography of Mono- and Oligosaccharides Derivatized withP-Aminobenzoic Ethyl Ester on a C18-Bonded Silica Column

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Abstract
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°C. Sugar-ABEE derivatives are separated well within a short run time(ca. 25 min) by reverse-phase partition chromatographic mode, and CIS-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. β-Anomeric configuration of [Glc-β-(1→4)Glc]-oligomer has shorter retention time than α-anomer in RP-HPLC. Column temperature and ionic strength slightly affect the elution behavior of sugarABBEE derivatives.