Abstract

A liquid chromatographic method has been developed and validated for the determination of the of Torsemide and Amiloride hydrochloride. Effective chromatographic separation was achieved on an eclipse Enable -C18 (4.6 mm X 250 mm, 5µm) column using gradient reverse phase technique. The mobile phase employed was ACN: 50Mm Potassium phosphate buffer (40:60), the pH was adjusted to 3.0 by ortho phosphoric acid. The flow rate was maintained at 1.0 mL/min and elute was monitored at 288nm. A linear response was observed over the concentration range 2-12µg/mL (R2=0.999) of AmilorideHCl and the concentration range 2-12 µg/mL (R2=0.998) of Torsemide. The limit of quantitation (LOQ) and limit of detection (LOD) for Torsemidewere 0.007 and 0.02 µg/mL, respectively and for AmilorideHClwere 0.02 and 0.07 µg/mL, respectively. The method was successfully validated in accordance to ICH guideline Q2. The RSD for intra-day and for inter-day precision were found to be lesser than 1.5% forAmilorideHCl. The RSD for intra-day and for inter-day precision were found to be lesser than 1.7% for Torsemide. The percentage recovery was found to be greater than 98.0 %.The method was found to be accurate, precise, linear, specific, sensitive, rugged, robust and stability indicating.