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Highly Enantioselective HPLC Separation of 5-Methyl-5-phenylhydantoin on the Norvancomycin-bonded Chiral Stationary Phase1 Yang Yiwen, Su Baogen, Shan Hanwen, Ren Qilong Institute of Pharmaceutical Engineering, Department of Chemical and Biochemical Engineering Zhejiang University, Hangzhou, China (310027) E-mail ：ceywyang@ Abstract Enantiomers of 5-Methyl-5-phenylhydantoin (MPH) were separated on norvancomycin chiral stationary phase (NVC-CSP), which was prepared by covalently linking norvancomycin to acid-treated silica gel. Three mobile phase modes have been investigated: reversed phase, polar organic mode, and normal phase. MPH may be enantioseparated under the reversed phase and polar organic mode. There is no enantioseparation under the normal phase. The effects of composition, column temperature, pH, and flow rate of mobile phase on the enantioseparation were investigated. The enantioseparation by methanol/2% TEAA (pH 5.5) 10/90 at the flow rate 0.6 ml/min and 20ºC is excellent. The hydrophobic interactions and hydrogen bonding are the dominant interactions on enantioseparations between the analyte and macrocycle in this chromatographic system. Keywords: 5-Methyl-5-phenylhydantoin; Norvancomycin; Enantioseparation; Chiral stationary phase 1. Introduction Enantioseparation has increasingly become important, especially in the pharmaceutical and food industries as optical isomers often possess different biological properties. The analysis and preparation of a pure enantiomer usually involves its resolution from the antipode. Among all the chiral separation techniques, HPLC has proven to be th