A sensitive and enantioselective method for the determination of the dihydropyridine calcium antagonist manidipine in human plasma was developed by high performance liquid chromatography (HPLC). The method involves a rapid and simple extraction in human plasma based on C8 bonded-phase extraction, and extraction samples were purified and concentrated on the C1 pre-column by using the column-switching technique.(+)-Manidipine, (−)-manidipine and (+)-barnidipine as an internal standard were detected by ultraviolet detection at 254 nm, and enantiomers of manidipine were quantitatively separated by HPLC on a Sumichiral OA-4500 column, containing chemically modified Pirkle-type stationary phase. Calibration graphs were linear for each concentration of enantiomer from 1.0 to 25 ng/mL and the detection limit in human plasma was 0.5 ng/mL for each enantiomer. The recoveries of (+)-manidipine and (−)-manidipine added to plasma were 93.6%-99.3% and 91.5%-99.1%, respectively, with coefficients of variation of less than 9.5%, respectively.