Separation and analysis of 4'-epimeric UDP-sugars, nucleotides, and sugar phosphates by anion-exchange high-performance liquid chromatography with conductimetric detection

Abstract

An anion-exchange HPLC method coupled with conductimetric detection was developed for the analysis of UDP-sugars, nucleotides, and sugar phosphates, each in a single chromatographic run. The analysis was applicable to concentrations over 50 pmol. The utility of this technique was demonstrated by the measurement of UDP-sugar 4'-epimerase activity of cell-free extracts from wild-type and mutant Neisseria meningitidis serogroup B strains. Additionally, the method has been applied to the analysis of the intermediates of glycolysis in human erythrocytes.