The interaction of glucose with pyrite has been investigated by a series of surface analyses, such as amounts adsorbed, TG, FTIR and XRD measurements. The adsorption experiment reveals that glucose would rapidly adsorb on the pyrite surface within 60min. However, physical adhesion characterized as multi-molecular layer adsorption may occur based on adsorption isotherm type. All results obtained by TG and FTIR suggest that no adsorption took place after several times of washing. XRD results indicate that crystal structure of pyrite was not obviously destructed after glucose was added. The interplane distances of (111), (400) after interaction between glucose and pyrite were widened, which showed the similar manner with that of before interaction. All these convincing data imply that physical adsorption predominantly governs the interaction of glucose on pyrite.