DEVELOPMENT OF RP-HPLC METHOD FOR SIMULTANEOUS DETERMINATION OF NEBIVOLOL HYDROCHLORIDE, VALSARTAN AND HYDROCHLOROTHIAZIDE IN CO-ADMINISTRATED ANTIHYPERTENSIVE AGENTS AND HUMAN PLASMA

Abstract

Reversed phase high performance liquid chromatography combined with photodiode array (HPLC-PDA) detector method was developed for simultaneous determination of two binary antihypertensive combinations; either Nebivolol (NEB) with Hydrochlorothiazide (HCZ) or with Valsartan (VAL) in pharmaceutical formulation and spiked human plasma. The method was successfully applied to separate the three drugs on one chromatogram under the same chromatographic conditions that used for two binary combinations. The separation was established on Xterra C18 column using gradient mobile phase which composed of acidified water of pH = 3.0 (adjusted with 0.1% orthophosphoric acid) and acetonitrile with flow rate of 1.0 mL/min. The drugs were determined at 280 nm. Concentration range of each analyte was shown linearity at 5.0–70.0 μg/mL for NEB, 8.0–60.0 μg/mL for HCZ and 5.0-80.0 μg/mL for VAL. The results of method validation were displayed according to ICH guidelines. The green assessment of the proposed has been carried out using two different tools.