This paper aims to establish a chiral separation method for miconazole using high performance liquid chromatog. (HPLC) on Chiralpak AD-H chiral column. The influences of the concentration of organic solvents and flow rate of the mobile phase, and temperature were examined Miconazole could be separated optimally on Chiralpak AD-H chiral column using n-hexane-alc. (volume:volume= 90:10) as mobile phase at a flow rate of 0.6 mL/min-1 and at 25°. The method is simple, quick, and reproducible. It can be used for enantioseparation of miconazole.