Development and Validation of RP-HPLC Method for the Simultaneous Determination of Amiloride Hydrochloride and Furosemide in Pure and Pharmaceutical Dosage Form

A simple, selective, rapid, precise and economical reverse phase high performance liquid chromatographic method has been developed for the simultaneous estimation of Amiloride Hydrochloride and Furosemide from pharmaceutical formulation. The method was carried out on an Enable C18 (250x4.6) mm; 5 μm with a mobile phase consisting of acetonitrile: Water (adjusted to pH 4 using orthophosphoric acid) in the ratio of 70:30 v/v at a flow rate of 1.0 ml/min. Detection was carried out at 281nm. The retention times of Amiloride Hydrochloride and Furosemide were 2.21 min. and 7.60 min., respectively. The developed method was validated according to ICH guidelines for evaluation of accuracy, precision, linearity, limit of detection, limit of quantitation and robustness. The proposed method can be used for the estimation of these drugs in combined dosage form.