Determination of monosaccharides and sugar alcoholsin tissues from diabetic rats by high-performance liquid chromatography with pulsed amperometric detection

Abstract

A sensitive and simple high-performance liquid chromatographicmethod has been developed to determine the concentration of monosaccharides and sugar alcohols in animal tissues. Five neutral monosaccharides ( d-glucose, d-galactose, d-mannose, d-fructose, and d-ribose) and three neutral sugar alcohols (myo-inositol, glycerol, and d-sorbitol) predominate in the renal cortices and sciatic nerves of rats. These monosaccharides and sugar alcohols were extracted with distilled water, purified by deproteinization with ethanol, a Sep-Pak C 18 cartridge, and columns of Dowex 50W-X8 and Amberlite CG-400, then separated on Ca 2+ and Pb 2+ cation-exchange columns, eluted with deionized distilled water at 80°C, and detected using integrated pulsed amperometry. About 10 pmol of each sugar was detectable with a signal-to-noise ratio of 10:1. d-Glucose, d-fructose, d-sorbitol, and d-mannose were higher in both the renal and sciatic tissues of diabetic rats than in those of normal animals. d-Ribose and glycerol were higher in the renal cortex of diabetic animals.