Abstract

DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR THE DETERMINATION OF ATENOLOL IN HUMAN PLASMA

Ahmed Yusuf, Syed N. Alvi and Muhammad M. Hammami*

ABSTRACT

A simple, rapid and precise reversed phase HPLC assay for the determination of atenolol in human plasma was developed and validated. The method used 0.25 ml of human plasma and a simple liquid/liquid extraction procedure. The chromatographic separation of atenolol and nadolol (internal standard, IS) was achieved on Symmetry Shield RP18 (5-μm 4.5 x 150-mm column) at room temperature. The mobile phase was composed of 10 mM sodium hydrogen phosphate containing 7.3 mM sodium lauryl sulfate (pH= 3), methanol, and acetonitrile (40:57:3, v:v:v), delivered at a flow rate of 1.0 ml/min. The eluted components were detected by a fluorescence detector set at the excitation and emission wavelengths of 229 nm and 298 nm, respectively. Under these conditions, no interference was observed and the retention times of atenolol and the IS were around 5.4 and 8.3 min, respectively. The relationship between atenolol concentration in plasma and peak height ratio of atenolol to the IS was linear (R2 ≥ 0.9992) in the range of 0.01 – 1.5 μg/ml. The intra- and inter-day coefficient of variations were ≤ 5.3% and 5.2%, respectively. Mean extraction recovery of atenolol and the IS from plasma samples was ≥ 76% and 87%, respectively. The method was used to assess the stability of atenolol in human plasma under various clinical laboratory conditions.

Keywords: Atenolol, Human Plasma, Nadolol, HPLC.