Development and validation of stability indicating RP-HPLC method for Simultaneous determination of Hydrochlorothiazide and Nebivolol Hydrochloride in pharmaceutical dosage form

Abstract

A stability-indicating RP-HPLC method has been established for determination of Hydrochlorothiazide (HCTZ) and Nebivolol hydrochloride (NBL) in combine dosage form under different stress conditions. The drug substances  were  subjected  to  stress  by  hydrolysis  (0.1N  HCl  and  0.5NaOH),  photochemical,  and  UV degradation (254 nm). Both the drugs were degraded under selected experimental conditions. Successful separation of the drugs from the degradation products was achieved on a  Hypersil BDS C18, 150x4.6, 5µ column  with  50:50 (v/v)  Mixed buffer and Methanol in the ratio 50:  50.    2.6 gms of potassium hydrogen phosphate and 0.6gms of dipotasssium hydrogen phosphate were dissolved in 1000 ml distilled water, pH was adjusted 5.5±0.1 with dilute orthophosphoric acid as mobile phase. The method was linear over the concentration range of 6.250-38.750 μg mL-1  (r2 > 0.991), with limits of detection and quantitation (LOD and LOQ) of 0.3061 and 0.9276 μg mL-1, respectively, for HCTZ and 2.500-15.000 μg mL-1  (r2  > 0.991) with LOD and LOQ of 0.489  and  1.4815  μg  mL-1,  respectively,  for  NBL.  The  method  was  validated  for  specificity,  selectivity, accuracy,  and  precision.  This  method  was  reproducible  and  selective.  As  the  developed  method  could effectively separate the drugs from their degradation products, it can be used as stability -indicating.