Simultaneous analysis of a series of phosphorylated sugars in small tissuesamples by anion exchange chromatography and pulsed amperometric detection

A simple method was developed to analyse a series of phosphorylated sugars in small physiological samples with dry tissue weights as low as 10 mg. Pho sphorylated compounds were extracted from the tissue after boiling in 80% m ethanol by re-suspension of the dried methanol phase in water. The sugar ph osphates were separated by Dionex high performance liquid chromatography an d detected by pulsed amperometry, in contrast to the common protocol for th e separation of sugars, the sugar phosphates were eluted at a low temperatu re and at a relatively low concentration of hydroxide in order to preserve the stability of the nucleotide sugars ADP-glucose and UDP-glucose. At low concentrations, the separation, detection and recovery of these compounds w as highly satisfactory and an almost linear relationship between concentrat ion and response was obtained. Quantities as low as 0.1 nmol could easily b e detected. Copyright (C) 1999 John Wiley & Sons, Ltd.