Abstract
Purpose: To develop a sensitive, simple and validated high performance liquid chromatography (HPLC) analytical method for the determination of metoprolol tartrate in rabbit plasma. Methods: Mobile phase of methanol and 50 mM ammonium dihydrogen phosphate solution (50:50) at pH 3.05 was used for separation of metoprolol on BDS hypersil C18 column at a wavelength of 223 nm. Flow rate and retention time were 0.6 mL/min and 7.4 min, respectively. For pharmacokinetic study, rabbits were given an oral dose of 8 mg/kg of metoprolol in solution form. Blood samples were taken from jugular vein of the rabbits after drug administration and analysed by HPLC. Results: Separation of metoprolol was not interfered with other components in plasma. The calibration curve was linear in the range of 25 - 1000 ng/mL (r 2 = 0.997). Lower limits of detection (LLOD) and quantitation (LLOQ) were 8.87 and 25 ng/mL, respectively. Relative standard deviation (RSD) of intraday and inter-day precision was < 14.27 and 7.61 %, respectively. Relative error of accuracy was between 4.85 and 14.37 %. Maximum plasma concentration (C max ), time to maximum plasma concentration (T max ) and half-life (t½) after metoprolol oral administration in rabbits were 186.29 ng/mL, 0.50 h and 2.27 h, respectively. Conclusion: A simple, accurate and precise HPLC-UV method for metoprolol determination in rabbit plasma has been successfully developed and applied to a pharmacokinetic study. Keywords: HPLC-UV, Metoprolol, Pharmacokinetics, Rabbit plasma, Liquid-liquid extraction, Validation
Highlights
Metoprolol tartrate is a selective β1-adrenergic blocking agents used for management of angina pectoris and treatment of mild to moderate hypertension
Several high performance liquid chromatography (HPLC) methods are available for Metoprolol quantification in plasma, urine and pharmaceuticals [3,4,5,6]
The mobile phase consisting of methanol and 50 mM ammonium dihydrogen phosphate was selected because of symmetry of drug peak, sensitivity and reasonable retention time
Summary
Metoprolol tartrate is a selective β1-adrenergic blocking agents used for management of angina pectoris and treatment of mild to moderate hypertension. Metoprolol is absorbed up to 95 % but undergoes intensive first pass metabolism after oral administration with bioavailability of 40 to 50 %. Present HPLC method utilizes UV detector as it is available, affordable and convenient and requires less maintenance [7,8,9,10]. Liquid-liquid extraction, solid-phase micro-extraction and solid-phase extraction are commonly used for the preparation of biological samples Most of these methods require various steps for the extraction of metoprolol from blood samples [11,12]. Aim of present study was to develop a sensitive, precise, accurate and simple HPLC-UV method for metoprolol determination in plasma and investigate its application for pharmacokinetic study in rabbits
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