Determination of Reducing Sugars by High Performance Liquid Chromatography with Postcolumn Fluorescence Derivatization Using 1, 2-Bis(4-methoxyphenyl)ethylenediamine

Abstract

A sensitive method for the determination of reducing sugars is described which employs high performance liquid chromatography with on-line postcolumn fluorescence derivatization. Reducing sugars are separated by borate complex anion-exchange chromatography on a trimethylammonium-bonded resin column (TSK gel Sugar AXG) and automatically converted into fluorescent derivatives by reaction with 1, 2-bis(4-methoxypheny)ethylenediamine in an alkaline medium. The detection limits (S/N=3) for reducing sugars are 30-160pmol in a 100-μl injection volume. The quantification of reducing sugars in human urine and serum is also described.