Development and Validation of RP-HPLC Method using UV detection for Simultaneous quantification of Amlodipine besylate and Nebivolol hydrochloride in Fixed-dose combination tablets

Abstract

Calcium channel blockers and beta blockers are the class of choice of drugs in treatment of hypertension and cardio vascular diseases. Such a fixed dose combination is viable to deal with the patients for whom monotherapy is not sufficient. A simple, responsive and productive high performance liquid chromatography method was designed and approved for simultaneous quantification of Amlodipine Besylate (AMB) and Nebivolol Hydrochloride (NBL) in tablet dosage form. In the developed method mobile phase comprised of a mixture of aqueous buffer (pH 6) and acetonitrile (55:45 v/v, respectively) at a flow rate of 1.5 ml/min. Chromatographic separation was achieved using Reverse-phase Kromasil 60-5-CN column, 250 × 4.6 mm (particle size 5 μm). Method was discovered specific as no obstruction of excipients peaks at the retention time of tablets was observed and demonstrated linearity in concentration range of 30.4-70.8 mcg/ml for AMB and 29.8-69.4 mcg/ml for NBL with R2> 0.999. Accuracy values for AMB and NBL was found to be 98.21-99.49% and 97.91-99.09%, respectively. Inter-day and intra-day precision was carried out and % RSD was found to be less than 2%. Robustness was performed to evaluate the results with slight variations in method and % RSD found to be less than 2%. Solution stability studies were conducted in normal conditions. Force degradation study was carried out using Photo-diode Array (PDA) detector and revealed that acid degradation in AMB and NBL was 7.12% and 1.88%, alkaline degradation was 9.61% and 0.05% and oxidative degradation was 8.50% and 1.59%, respectively.