Development and validation of HPTLC and Green HPLC methods for Determination of Furosemide, Spironolactone and Canrenone, in Pure forms, Tablets and Spiked Human Plasma

Research Abstract

Two selective and accurate chromatographic methodsare presented for simultaneous quantitationof furosemide (FR), spironolactone (SP) and canrenone (CN); the main degradation product and the mainactive metabolite of spironolactone. Method (A) is HPTLC,where separation was completedon silica gel HPTLC F254platesusing ethylacetate: triethylamine: acetic acid (9: 0.7: 0.5, by volume) as a developing systemand UV detectionat254 nm. Method (B) is agreen isocratic RP-HPLC utilizing C18 (4.6 x 100 mm) column, themobile phase consisting of ethanol: deionized water (45: 55, v/v) and UV estimation at 254 nm.Adjustment of flow rate at 1 ml/min and pH at 3.5 with glacial acetic acid was done.Regarding greenness profile, the proposed RP-HPLC method is greener than the reported one. ICH guidelines were followed to validate the developed methods. Successful applications of the developed methods were revealed by simultaneous determination of FR, SP and CN in pure formsand plasmasamples in ranges of 0.2-2 µg/band, 0.05-2.6 µg/band and 0.05-2 µg/band for method (A) and 5-60 µgmL-1, 2-60 µgmL-1 and 2-60 µgmL-1 for method (B) for FR, SP and CN, respectively.

Research Keywords