Rapid Separation of Five Anti-Hypertensive Agents - Atenolol, Metoprolol, Hydrochlorothiazide, Amlodipine and Nebivolol: Application to Estimation of Metoprolol Succinate in Tablet Dosage Form
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.
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