Abstract: By adopting the bifunctional group reagent methylene-di-paraphenylene iso-cyanate　(MDI), a ring antibiotic chiral stationary phase (CSP) was prepared in non-aqueous dimethylfomamide (DMF) with vancomycin which is a macrocyclic glycopeptide antibiotics bonded on γ-amidopropyl silica gel. It was used in the chiral separation in high performance liquid chromatography. The results showed that vancomycin CSP synthesized in this way can be well used in enantiomer separation of the chiral compounds in normal and reversed-phase. It can be confirmed that the separation capability in reversed-phase is better than that in normal phase. Seventeen pairs of enantiomers were separated in reversed-phase mode, and the results showed a wide range of application. The phosphorus buffer system was better than the triethylamine-acetic acid buffer system. The resolution of some chiral drugs such as D,L-dansyl amino acid has certain rule, and provides information about absolute configuration. The aging or denaturalization can not happen in the CSP when phase system was transformed, so it has some stability. The mechanism of the separation was analyzed and found to be the same as that of Armstrong.

Key words: stationary phase, chiral separation , vancomycin, high performance liquid chromatography