Original Articles
P. Ravisankar, G. Devala Rao,
Abstract
One of the key goals of High Performance Liquid Chromatography technique is to achieve a consistent and reproducible separation. A simple, precise, selective and sensitive HPLC method with UV detection was developed for separation of five anti-hypertensive agents, atenolol hydrochloride, metoprolol succinate, hydrochlorothiazide, amlodipine besylate and nebivolol hydrochloride and validated for determination of metoprolol succinate. RPHPLC method was developed by using Welchrom C18Column (4.6 mm i.d. X 250mm, 5µm), Shimadzu LC-20AT ProminenceLiquid Chromatograph. The mobile phase composed of10 mM Phosphate buffer (pH3.0,adjusted with triethylamine): acetonitrile(50:50, v/v).The flow rate was set to 1.0 ml/min with the responses measured at 235nm using Shimadzu SPD-20A Prominence UV-Visible detector. The retention times of atenolol hydrochloride, Metoprolol succinate, hydrochlorothiazide, amlodipine besylate and nebivolol hydrochloride were found to be 2.303 min, 2.827 min, 3.500 min, 4.253 minand 4.957 min respectively. The separation was achieved within 6 min. The statistical validation of the developed method was carried out according to ICH guidelines. Metoprolol succinate was found to give linear response in the concentration range of 2-10µg/ml. Recovery studies were performed to ascertain the accuracy by standard addition method and average recovery was found to be 99.8-100.75%. The LOD and LOQ were found to be 0.1840 µg/ml and 0.5578 µg/ml respectively. The developed method can be used for routine quality control analysis of metoprolol succinatein pharmaceutical tablet dosage form. It can also be extended for the determination of other most commonly prescribed anti-hypertensive agents namely atenolol hydrochloride, hydrochlorothiazide, amlodipine besylate and nebivolol hydrochloride. This method provides a fast, simple method with excellent peak symmetry and high resolution.