Validated HPTLC Method Development for Simultaneous Estimation of Spironolactone and Hydrochlorothiazide in Formulation

Abstract

This study describes a new TLC method for simultaneous quantification of Spironolactone and Hydrochlorothiazide in bulk and tablet dosage forms that is simple, precise and accurate. The drugs were separated using aluminium precoated plate with silica gel 60 F-254 as the stationary phase using Toluene: Ethyl acetate: Methanol 6: 4: 1 (v/v/v) as the mobile phase. At 231 nm, the separated zones were densitometrically evaluated. The two drugs were satisfactorily resolved with RF values of 0.78 ± 0.02 and 0.33 ± 0.02 for Spironolactone and Hydrochlorothiazide. The accuracy and reliability of the method were assessed by evaluation of linearity (120-270 ng/spot for Spironolactone and 100-350 ng/spot for Hydrochlorothiazide), precision (intra-day RSD 0.98-1.80 % and inter-day RSD 0.76-1.92 % for Spironolactone, and intra-day RSD 0.84-1.85 % and inter-day RSD 0.31-1.68 % for Hydrochlorothiazide), accuracy (100.78 ± 0.49 % for Spironolactone and 98.54 ± 1.80 % for Hydrochlorothiazide, by ICH guidelines. These methods have been applied to formulation and no interference of excipients is seen which proves that the method is specific.