Abstract: As one of the most important biological macromolecules in the organism,glycoprotein participated in the recognition,adhesion and migration of cells,regulates the proliferation and differentiation of cells. A large number of studies had shown that glycoproteins were widely used in food-derived materials and exhibited a variety of physiological activities. However,because of the complexity of glycosylation,the analysis of glycosylation sites had become a crucial link in the characterization of glycoprotein structure. In this review,N-glycosylation,O-glycosylation and partial sugar chain structure of glycoproteins were introduced. N-glycosylation was set as the main research object,and two approaches for nuclear magnetic resonance(NMR)and mass spectrometry techniques were introduced. Mass spectrometry technique detection method was widely used because it was suitable for the analysis of mixtures. According to the research results of glycosylation sites from food-derived glycoprotein in recent years,two approaches were concluded. Subsequently,the methods of fragmentation,the mechanism of deglycosylation and the principles of glycosylation sites analysis based on glycopeptides were clarified. In order to provide reference for the determination of glycosylation sites by biological mass spectrometry,and paved a way for the in-depth study of the nature and function of food-derived glycoprotein.