Abstract: AIMTo establish new methods for the chiral separation and preparation of three new drugs, alfuzosin, terazosin and doxazosin. METHODSBy optimizing factors which affect the chiral separation, modifier of solvent, chiral additive, pH of mobile phase, modifier of organic base and stationary phase, the optimum condition for chiral separation were selected. The preparation of enantiomers was carried out on semi-preparative reverse phase column (7.8 mm×250 mm C₄ 5 μm). Acetonitrile-water modified by the addition of carboxymethyl-β-cyclodextrin (2%～5%, w/v) was applied as chiral mobile phase. RESULTSThe enantiomers of three new drugs were base-line-separated and milligram-scale samples of enantiomer were obtained. CONCLUSIONThe newly established method can be used in research and development of the enantiomers of three new drugs.