In this study, we provide a simple and straightforward method to distinguish core-fucosylated glycans from antenna-fucosylated glycans by systematically evaluating the extent of fucose migration based on the feature Y ions by using Fut8 knockout (Fut8−/−) mouse brain under HCD energy of 20%. The method was further evaluated by multiple different approaches including validation by standard glycopeptides, evaluation in silico, comparison under the lower energy (15%), and applied to analyze core-fucosylated glycans in wide-type (Fut8+/+) mouse brain.