The invention discloses an analysis and test method of puraconazole optical isomer. The method includes: adopting liquid chromatography, wherein solid phase of liquid chromatography is octadecyl silane bonded silica gel while mobile phase of the same is acetonitrile water solution, voluem percentage concentration of acetonitrile is 30-50%, 4-tert-butyl cyclohexanol at effective concentration is contained in the acetonitrile water solution, and 4-tert-butyl cyclohexanol is a cis-trans isomer mixture. By using the method, effective separation of the puraconazole optical isomer can be realized on a common C18 chromatographic column, so that application cost is lowered remarkably, and chromatographic column consumable cost can be saved greatly.