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Original Article
Year : 2017   |  Volume : 5  |  Issue : 3  |  Page : 23-27

Method development and validation of hydrochlorothiazide and Nebivolol in bulk and tablet formulation by reverse phase-high performance liquid chromatography method

The present work describes development and validation by high-performance liquid chromatography by UV detector procedure for the analysis of hydrochlorothiazide (HCZ) and Nebivolol (NBL) in pharmaceutical mixture. Effective chromatographic separation of HCZ and NBL was achieved using a Youngline (S.K.) Gradient System UV detector. Reverse phase (GRACE) C18 column, (4.6 mm × 250 mm; 5 μm), an SP930D pump, a 20 μl injection loop, and UV 730D absorbance detector and running Autochro-3000 software. Mobile phase composed of 0.1% solution of orthophosphoric acid in water and methanol in the proportion of (40:60), respectively. The flow rate is 0.7 ml/min on detecting wavelength 282 nm. The PH of mobile phase was 3.2. The proposed high-performance liquid chromatography method was statistically validated with respect to linearity, ranges, precision, accuracy, selectivity, limit of detection, limit of quantitation and robustness. The retention time of HCZ and NBL and were found to be 4.6833 and 9.18333, respectively. All parameters were found to be within the acceptance limit. The calibration curves were linear in ranges of 25-125 and 10-50, μg/ml respectively for HCZ and NBL. The R2 of HCZ and NBL was found to be 0.999, 0.999, respectively.
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