ISOCRATIC SEPARATION OF FOUR BETA BLOCKERS WITH AMLODIPINE BY C18 RP-HPLC: APPLICATION TO AMLODIPINE DETERMINATION IN PHARMACEUTICAL DOSAGE FORMS
Journal Title: International Research Journal of Pharmacy (IRJP) - Year 2013, Vol 4, Issue 6
Abstract
This method described for the successful separation of a beta blockers with amlodipine by RP-HPLC on a C18 column with UV detection. 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 was developed and validated for determination of five anti-hypertensive agents, atenolol hydrochloride, metoprolol succinate, propranolol hydrochloride, amlodipine besylate and nebivolol hydrochloride with application to estimation of amlodipine besylate. RP-HPLC method was developed by using Welchrom C18 column (4.6 mm i.d. X 250mm, 5µm), Shimadzu LC-20AT ProminenceLiquid Chromatograph. The mobile phase composed of 10mM Phosphate buffer (pH3.0, adjusted with triethylamine): acetonitrile (50:50, v/v). The flow rate was set to 1.0ml/min with the responses measured at 235nm using Shimadzu SPD-20A Prominence UV-Visible detector. This method provides effective and reproducible separation of five anti-hypertensive agents less than 6 minutes. The retention times of atenolol hydrochloride, metoprolol succinate, propranolol hydrochloride, amlodipine besylate and nebivolol hydrochloride were found to be 2.310 min, 2.830 min, 3.473 min, 4.260 min and 4.960 min respectively. The statistical validation of the developed method was carried out according to ICH guidelines. Amlodipine besylate 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 99.83-100.40%. The LOD and LOQ were found to be 0.2251µg/ml and 0.6823µg/ml respectively. The developed method can be used for routine quality control analysis of amlodipine besylate in pharmaceutical tablet dosage form. It can also be extended for the determination of other above mentioned most commonly prescribed anti-hypertensive agents. The analysis yields a simple, rapid analysis, reduction in runtime, with excellent peak shape, high resolution as well as outstanding reproducibility.
Authors and Affiliations
Ravisankar Panchumarthy , Garikapati Rao
Keywords
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- EP ID EP93370
- DOI 10.7897/2230-8407.04620
- Views 136
- Downloads 0
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