Abstract: Glycosylation is a common form of protein post-translational modification and plays key roles in biological processes, which is closely related to the occurrence, development, diagnosis and treatment of various diseases. Although the rapid development of biological mass spectrometry-based technology offers great conveniences to identification of glycosylation sites, peptides sequences and glycan structures of glycoproteins respectively, the illumination of the linkage between proteins and glycans and the analysis of the structure of intact glycopeptides still face a lot of difficulties and challenges. Based on literature researches, the major directions of the field were determined. This review will focus on MS analysis of the site-specific intact glycopeptides and describes the development and prospects of biological mass spectrometry-based technology for the analysis of intact glycopeptides on many aspects, from ion source and mass analyzers to dissociation modes and the analysis of mass spectrometric data.