Microquantitative analysis of neutral and amino sugars as fluorescent pyridylamino derivatives by high-performance liquid chromatography.
A method for the quantitation of picomole amounts of neutral and amino sugars in glycoconjugates was developed. Glycoconjugates were hydrolyzed with a mixture of equal amounts of 4 M trifluoroacetic acid and 4 M hydrochloric acid, and the free amino groups were acetylated. Sugars were coupled with 2-aminopyridine. After the excess reagents were removed by gel-permeation high-performance liquid chromatography, the fluorescent pyridylamino derivatives of sugars were separated and quantified by high-performance liquid chromatography on a reversed-phase column. This method allowed the determination of 0.01-10 nmol of sugars. About 100 pmol of several glycoconjugates were analyzed by the present method, with satisfactory results.
D000606:Amino Sugars / Q000032:analysis*; D000631:Aminopyridines; D002241:Carbohydrates / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D006868:Hydrolysis; D008832:Microchemistry; D013050:Spectrometry, Fluorescence
D000606:Amino Sugars; D000631:Aminopyridines; D002241:Carbohydrates; C032439:alpha-aminopyridine