Separation of partially methylated mannitols by liquid chromatography

To facilitate the methylation linkage analysis of complex carbohydrates containing radioactively labeled mannose residues, an HPLC micromethylation technique has been developed which effectively resolves all of the partially methylated mannitol standards prepared using an under-methylation protocol. The method was applied to the linkage analysis of the nine mannose residues of the dolichol-derived oligosaccharide Glc3Man9GlcNAc2 isolated from BHK-21 fibroblasts. This technique should prove widely applicable to the methylation linkage analysis of radiolabeled complex carbohydrates.