Development of microwave-assisted methods to aid in carbohydrate analysis

This study involved development of three high-throughput chemical techniques to assist in carbohydrate analysis. The methods utilized both microwave radiation and a volatile organic base to achieve short reaction times and high product yield. Two methods, one reductive and one non-reductive, released intact O-glycans for structural analysis, and one method aided in determination of the site of O-glycosylation on the peptide. Optimal reaction conditions for each method were determined on standard glycoproteins. In comparison to common release methods, microwave-assisted reductive O-glycan release provided higher yields of O-glycans in every case. O-glycans were released rapidly and were able to be subsequently labeled for chromatographic separations by the non-reductive release method. The O-glycosylation site was determined following release by the formation of peptide-dimethylamine adducts, which, coupled with the addition of a deuterium label further aided in selection of glycopeptides for analysis