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

Abstract

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.