Development and validation of a high performance liquid chromatography method for determination of 6‐benzyl‐1‐benzyloxymethyl‐5‐iodouracil (W‐1), a novel non‐nucleoside reverse transcriptase inhibitor and its application to a pharmacokinetic study in rats

A sensitive and selective high‐performance liquid chromatographic (HPLC) method for determination of 6‐benzyl‐1‐benzyloxymethyl‐5‐iodouracil (W‐1), a novel non‐nucleoside reverse transcriptase inhibitor in rat plasma, was developed and validated. Chromatographic separation of W‐1 and megestrol acetate (internal standard) was achieved on a reversed‐phase C 18 column at 25°C. The mobile phase was consisted of acetonitrile–water (60:40, v/v) and pumped at a flow rate of 1.0 mL/min. The ultraviolet (UV) detector was set at the absorption wavelength of 284 nm. The calibration curve for W‐1 was linear over the concentration range of 0.01–8 µg/mL and the lower limit of quantification was 10 ng/mL. The intra‐ and inter‐day precision and accuracy were <8.9 and 5.3%, respectively. The extraction recoveries ranged from 97.9 to 101.6%. The validated HPLC method was successfully applied to a pharmacokinetic study of W‐1 in rats. Copyright © 2015 John Wiley & Sons, Ltd.