Abstract:
In this work simple, precise and accurate spectrophotometric and high performance liquid chromatography (HPLC) assay methods for spironolactone in raw material and tablet dosage form were developed and validated. HPLC analysis method was developed using inertstil C18 (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. In spectrophotometric analytical method, two methods in UV region were developed and validated as Hydrazine and Hydroxylamine methods. The maximum absorption wavelength for determination of spironolactone was found to be 236.8 nm, but those for Hydrazine and Hydroxylamine derivatization were found to be 251.5 nm and 263.7 nm, respectively. Beer’s law was obeyed in the concentration range from 20 ppm to 30 ppm for both HPLC and spectrophotometric analysis methods. The assay percentage (mean ±RSD) for tablet commercial formulation was found to be (98.00±0.39) %, (97.15±0.28) % and (98.25±0.59) % for Hydrazine, Hydroxylamine and HPLC methods, respectively. The percentage of recovery was found to be (100.20-102.65) %, (98.47-100.67) % and (99.40-101.99) % for Hydrazine, Hydroxylamine and HPLC methods, respectively. The limit of detection (LOD), however, was found to be in the range from 0.028 ppm to 0.553 ppm for all methods. The limit of quantitation (LOQ) was also found to be in the range from 0.086 ppm to 1.677 ppm for all methods. All the validation methods were carried out according to ICH Q2 (R1) guideline.
