A quantitative determination by capillary gas-liquid chromatography of neutral and amino sugars (as O-methyloxime acetates), and a study on hydrolytic conditions for glycoproteins and polysaccharides in order to increase sugar recoveries

Abstract

Complete gas-liquid chromatographic separation of O-methyloxime acetates (syn and anti isomers) prepared from eight neutral sugars, three hexosamines, and muramic acid has been obtained, using a fused-silica Carbowax 20M capillary column. A single hydrolytic step for carbohydrate-containing biological material (less than or equal to 2.5 X 10(-3) M sugar solution in 4 N trifluoroacetic acid at 125 degrees C for 1 h) has been developed, and results have been compared with those obtained with standard hydrolytic conditions in order to ensure complete release of amino sugars from glycoproteins, together with minimum losses of neutral sugars. The combination of this acid hydrolysis with the above improved derivatization procedure for the gas-liquid chromatographic analysis has led to a simple, rapid, and sensitive analytical method, which has been successfully tested on three glycoproteins (fetuin, mucin, and peroxidase) and two plant cell-wall polysaccharide fractions (soluble fibers from carrots and soybeans).