Simultaneous determination of irbesartan and hydrochlorothiazide in human plasma by liquid chromatography

A simple, selective, sensitive and precise high-performance liquid chromatographic plasma assay for the antihypertensive drugs, irbesartan and hydrochlorothiazide is described. Good chromatographic separation was achieved using a Supelcocil C18 (5 μm, 15 cm×4.6 mm) column and a mobile phase consisting of 10 mM potassium dihydrogen phosphate:methanol:acetonitrile (5:80:15 v/v/v) (pH:2.5) while at a flow-rate of 1.0 ml min−1. Irbesartan and hydrochlorothiazide were detected at 275 nm and were eluted 5.8 and 7.8 min, respectively, after injection. No endogenous substances were found to interfere. The method utilizes protein precipitation with acetonitrile as the only sample preparation involved prior to reversed-phase high-performance liquid chromatography. No internal standard was required. Linearity range for irbesartan and hydrochlorothiazide was 10.0–60.0 μg ml−1 and 4.0–20.0 μg ml−1, respectively. The determination of intra- and inter-day precision (RSD) was less than 2.5 and 3.5%, at all concentration levels, while the inter- and intra-day accuracy (% difference) was less than 4.9–6.2%. This method is being used in a therapeutic drug monitoring service to quantitate these therapeutic agents in patients for pharmacokinetic studies.