Automated analysis of hexosamines by high-performance liquid chromatography with photometric and fluorimetric postcolumn labeling using 2-cyanoacetamide.
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.
D000328:Adult; D000818:Animals; D001322:Autoanalysis; D002851:Chromatography, High Pressure Liquid; D006595:Hexosamines / Q000032:analysis*; D006801:Humans; D006868:Hydrolysis; D008297:Male; D009570:Nitriles; D010777:Photochemistry; D013050:Spectrometry, Fluorescence
D006595:Hexosamines; D009570:Nitriles; C031594:2-cyanoacetamide