Abstract: A method was developed in determining 4 kinds of glucides which were fucose, galactose, α-methyl mannoside and N-between starch bioadhesive materials and sugar residues by high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS). The sample was filtrated and analyzed by HPLC-MS/MS. Analyte was separated by liquid chromatography with a mobile phase consisting of V(acetonitrile)∶V(water)=80∶20. The mass spectrometer was operated at the negative ion mode via electrospray ionization (ESI) source. The quantitative analysis was determined by the multiple reactions monitoring(MRM). Linear relationship between values of peak area of detection signals and concentration of fucose are obtained in the ranges of 1.0—40.0 mg•L^-1, while relationships of galactose, α-methyl mannoside and N-acetylglucosamine are obtained in the range of the linear ranges 1.0—80.0 mg•L^-1. The recovery rate of fucose is 111%, and the RSD is 6.55%. The recoveries rates of galactose, α-methyl mannoside and N-acetylglucosamine are 106.5%, 99.0% and 91.0%, the RSDs are 7.78%, 6.64% and 5.68%, respectively. Limits of detection (S/N≥3) for fucose, galactose, α-methyl mannoside and N-acetylglucosamine are 0.1, 1.0, 01 and 0.5 mg•L^-1, respectively. The method is proved to be rapid, reliable and sensitive, which is suitable for estimating the specific linking between starch bioadhesive materials and glucides. The proposed method is also suitable for rapid determination of glucides components in other complex system.