An improved method for preparation of perbenzoylated ganglioside‐derived sialic acids and nanogram detection ofN‐acetyl‐ andN‐glycolylneuraminic acid by high performance liquid chromatography

摘要

AbstractAn improved method for the preparation of perbenzoylated ganglioside‐derived sialic acids is described. After mild acid hydrolysis, isolation of sialic acids can be achieved by Folch partition (Method A) or by anion exchange chromatography (Method B). Perbenzoylated sialic acids were freed from benzoylation reagents by a second Folch partition. Total recoveries of both methods were found to be ⩾90, calculated from metabolically labelled gangliosides. DerivatizedN‐acetylneuraminic andN‐glycolylneuraminic acids were separated and quantified by isocratic high performance liquid chromatography using a RP18column as the stationary phase and methanol: water (8:2) as the mobile phase. Both sialic acids were completely separated and eluted as single peaks within 15 min, monitored by UV detection. As little as 20 ng of neuraminic acid could be detected, the detector being linear up to 5 μ