A rapid and simple high performance liquid chromatography (HPLC) mcthod wiih a UV detection (241 nm) was developed and validated for estimation of eplerenone from spiked human plasma. The analyte and the internal standard (valdecoxib) were extracted with a mixture of dichloromethane and diethyl ether. The chromatographic separation was performed on a HiQSil C-18HS column (250 mm × 4.6 mm, 5 um) with a mobile phase consisting of acetonitrile:water (50:50, v/v) at flow rate of 1 mL/min. The calibration curve was linear in the range 100 3200 ng/mL and the heteroscedasticity was minimized by using weighted least squares regression with weighting factor I/X.