A validated RP-HPLC method for the determination of Eplerenone in human plasma

A highly sensitive and accurate HPLC method was developed for the determination of Eplerenone in human plasma using Hydrchlorothiazide as internal standard. Diethyl ether was used as solvent in the liquid – liquid extraction process. A non-polar Phenomenex Prodigy ODS-2, C18 column (150 X 4.6 mm id) was chosen as the stationary phase and a binary mixture of 20 mM Sodium acetate buffer (pH 4.0 ± 0.05) and methanol in a ratio of 30:70 v/v was used as mobile phase. The drug and the internal standard were eluted under isocratic condition at a flow rate of 1.0 mL/min of the mobile phase and the wavelength of detection is 220 nm (UV – detector). The injection volume is 20 μL and the runtime of the method is 6 minutes. A good linearity was observed for the method over the range of 52.52 to 3089.48 ng/mL. The recovery of Eplerenone was 45.48 % with a standard deviation of 1.62 and recovery of internal standard was 75.32%. The LOD of Eplerenone was 52.52 ng/mL. Matrix effects were not observed