A new cyclodextrin-derived chiral stationary phase (denoted as CDA-CSP) was synthesized by immobilizing mono(6(A)-azido-6(A)-deoxy)-per(p-chlorophenyl carbamoylated) β-cyclodextrin derivative to alkynyl modified silica via click chemistry. This newly prepared CSP shows good enantioseparation performance for six chiral compounds (1-6), such as 4-phenyl-oxazolidine-2-thione, two kinds of aryl alcohols, substituted flavonoids and benzoin, in which baseline separation of Analytes 1-4 was achieved under the experimental conditions. The effects of column temperature, mobile phase pH and content of methanol on the enantioseparation characteristics of CDA-CSP were investigated in detail. Retention factor and resolution for Compound 3 gradually reduced with an increase of column temperature, and a good linear relationship was shown between napierian logarithm of selectivity factor and reciprocal of column temperature. In the pH range from 3.56 to 5.50, a change in pH hardly affected the resolution of Analyte 2. In addition, increasing methanol in the mobile phase resulted in rapid eluting of the analytes from the column in reversed-phase mode. The retention factors for Analytes 1 and 3 significantly decreased and their resolution showed different trends.