As a complementary approach to released glycan analysis, monosaccharide profiling allows analysts to verify glycan composition determined in traditional HILIC-based separations. A well-established assay for monosaccharide analysis uses reversed-phase chromatography to separate 2AA-derivatized monosaccharides.
In this application note, we demonstrate the ability of the ACQUITY UPLC H-Class Bio System to run both HPLC and UPLC methods, using monosaccharide standards as well as bovine fetuin and the commercially available monoclonal antibody cetuximab as analytes. This HPLC assay provided sufficient resolution of individual monosaccharides but was restricted in part by the time required to perform the separation. A significant reduction in runtime was obtained by transferring the legacy HPLC method to UPLC technology.
With the modernized, UPLC-based separation, a higher throughput assay for monosaccharide analysis was thereby achieved.