Development and Validation of High Performance Liquid Chromatography Assay Method of Spironolactone

Hozyfa A Hamdan and Ahmed E Mo

Abstract

A new, precise and accurate reversed-phase high performance liquid chromatograph a method was developed for determination of spironolactone. High performance liquid chromatograph was carried out by isocratic technique on a reverse-phase using C18 inertstil (250 *4.6 mm), 5μm column with a mobile phase consisting of phosphate buffer solution PH = 4 and acetonitrile in (1:1) ratio. The flow rate was adjusted at 1.5 ml/min, detection wavelength at 240 nm, temperature at 40 ᵒC and retention time was found to be 4.5 min. Beer's low was obeyed in concentration range 20-30 ppm. The assay percentage of spironolactone in tablet dosage form was found to be (98.25±0.59) %. The % of recovery was found to be (99.4-101.99) %. The limit of detection (LOD) and limit of quantization (LOQ) were found to be 0.553 ppm and 1.677 ppm respectively. The results of analysis have been validated statistically and recovery studies confirmed the accuracy of accuracy of developed method which was carried out according to ICH guidelines [3].

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