Automated analysis of hexosamines by high-performance liquid chromatography with photometric and fluorimetric...

Glucosamine and galactosamine were well separated in ca. 60 min on a Hitachi 2617 column (polystyrene sulfonate type, 4 mm X 25 cm) with a borate buffer (pH 7.5) containing sodium chloride. The hexosamines in the eluate were monitored fluorimetrically and photometrically at 331 (excitation)/383 (emission) and 276 nm, respectively, by postcolumn labeling with 2-cyanoacetamide. This simple method allowed the simultaneous, automated determination of 10-500 nmol of glucosamine and galactosamine with high reproducibility. This method was applied successfully to the analysis of hexosamines in some glycoconjugates.