Development and Validation of UV Spectrophotometric and RP-HPLC Methods for Simultaneous Estimation of Spironolactone Capsule Dosage Form

Abstract

Two simple, selective, accurate, precise, rapid and reproducible methods are developed and validated for the analysis of spironolactone and furosemide in their combined capsule dosage form. The first UV Spectrophotometric method involves Absorbance Correction method that includes measurement of absorbance at two wavelengths i.e. at 237 nm (λmax of spironolactone) and at 275 nm (λmax of furosemide) in methanol. Linearity was found in the range of 2.5-17.5 μg/ml and 1-7 μg/ml for spironolactone and furosemide, respectively. Correlation coefficient (r2) values obtained were 0.9994 for Spironolactone and 0.9993 for furosemide. The another method is based on separation of both these drugs by RP-HPLC using column Inertsil C18 ODS – 3V column (150 mm x 4.6 mm, 5 μ) in isocratic mode utilizing a mobile phase of methanol and phosphate buffer (pH 3.2) (60:40, v/v) at a flow rate of 1.5 ml/min with UV detection at 254 nm. The retention time of furosemide and spironolactone was found to be 2.52 min and 8.32 min, respectively. Linearity was found in the range of 25-75 μg/ml and 10-30 μg/ml for spironolactone and furosemide, respectively. Correlation coefficient (r2) values obtained were 0.9998 for spironolactone and 0.9999 for furosemide. Furthermore, the developed methods are validated in terms of various validation parameters according to ICH guidelines and can be used for routine analysis of the both the drugs in pharmaceutical formulation.