Abstract: In order to improve the stereoselectivity of the native β-cyclodextrin boned-silica gel stationary phase, 2,6-di-O-benzyl-β-cyclodextrin bonded stationary phase (BCDS) was prepared via the spacer γ-glycidoxypropyltrimethoxy-silane. The bonded silica was characterized by three methods (Fourier transform infrared spectrogram, Molish color reaction, X-ray optical electrical energy spectrogram). The chromatographic performances of BCDS were appraised by using toluene, dimethyl phthalate and phenanthrene. The selectivity of the new bonded-silica gel stationary phase was investigated by isomers. Some of the isomers such as nitroaniline, diphenols etc. were separated successfully by 2,6-di-O-benzyl-β-cyclodextrin bonded silica stationary. The results show that the introduction of benzyl to β-cyclodextrin leads to change of the retention of solutes and the dimensional selectivity.