Analysis of mono- and disaccharides by high-performance liquid chromatography of the benzyloxime-perbenzoyl derivatives

Abstract

A group of biologically important mono- and disaccharides are separated by high-performance liquid chromatography of the benzyloxime-perbenzoyl derivatives on a normal-phase microparticulate column with a hexane-dioxane mixture as the eluting solvent. A single, quantifiable derivative of each sugar is formed easily. These derivatives are detected by UV absorption at either 230 or 254 nm, with a sensitivity in the picomole range at the former wavelength. The sugars in the residues from the evaporation of small aliquots of biologic fluids (10-100 microliter) are derivatized without prior isolation and are determined quantitatively by the use of appropriate internal standards. The analyses could be performed routinely with a simple, inexpensive instrument.