Optimization and validation of a high-performance liquid chromatographic method with UV detection for the determination of pyrrole–imidazole polyamides in rat plasma

A simple and sensitive high-performance liquid chromatography (HPLC) method utilizing UV detection was developed for the determination of plasma pyrrole (Py)–imidazole (Im) polyamides in rats and applied to the pharmacokinetic study of compounds. After deproteinization of plasma with methanol, Py–Im polyamides were analyzed with a reversed-phase TSK-GEL ODS-80TM (4.6 mm × 15.0 cm TOSOH Co., Japan) column maintained at 40 °C. The mobile phase solvent A was 0.1% acetic acid and the solvent B was HPLC-grade acetonitrile (0–10 min, A: 100–20%, B: 0–80% linear gradient; 10–15 min, A: 40%, B: 60%). The flow rate was 1.0 ml/min. The detection wavelength was set at 310 nm. The method was used to determine the plasma concentration time profiles of Py–Im polyamides after intravenous injection.