BIOANALYTICAL METHOD DEVELOPMENT OF ATENOLOL IN RAT PLASMA BY ULTRA PERFORMANCE LIQUID CHROMATOGRAPHY AND TANDEM MASS SPECTROMETRY METHOD: APPLICATION TO PHARMACOKINETIC DRUG-HERB INTERACTION STUDY

Abstract

Objective: To develop a rapid, simple and sensitive ultra-performance liquid chromatography and tandem mass spectrometry (UPLC-MS/MS) method for quantitative estimation of atenolol (ATN) in rat plasma and its application to pharmacokinetic drug-herb interaction study.

Methods: Simple precipitation method was used for the extraction of plasma samples with an aliquot of 25 μl plasma samples extracted using acetonitrile precipitation technique containing internal standard. Chromatographic separation was performed using Phenomenex, Kinetex, C18, 50 x 2.1 mm, 1.7µ by a gradient mixture of 0.1 % formic acid in acetonitrile and 10 mmol Ammonium formate as a mobile phase at the flow rate of 0.7 ml/min. The analyte was protonated in the positive electrospray ionization (ESI) interface and detected in multiple reactions monitoring (MRM) modes using the transition m/z 145.0-267.2.

Results: The developed method had an advantage of fast chromatography run time of 1.8 min with improved sensitivity over existing methods and broader application to use for drug-herb or drug-drug interaction studies in rat pharmacokinetic. Calibration curves have been linear over the wide range of 4.93-5047.00 ng/ml in rat plasma which covers a wide range of plasma concentrations form study samples. ATN is widely used as a biomarker for determination of the drug-herb or drug-drug interactions studies, especially with the drugs that alter the permeability of another administered drug(s) and the developed method can be used to explore drug interaction studies. Especially to evaluate the role of concurrently administered drugs and its impact on permeability enhancer or absorption enhancer drugs following oral dose administration. This method will be significantly useful for the drug-herb pharmacokinetic interaction studies where we cannot quantify the herb because of multiple components in the hydro-alcoholic or aqueous extract of any herb, so choice remained is to quantify ATN and therefore it will be extremely helpful in such scenario.

Conclusion: It was concluded that the developed UPLC-MS/MS method was sensitive linear and rapid; can be used for quantification of ATN in rat plasma for pharmacokinetic drug-herb interaction study.