Xây dựng phương pháp định lượng itoprid trong huyết tương người bằng phương pháp HPLC với detector huỳnh quang

Abstract

A new method was developed for determination of itopride in human plasma by high-performance liquid chromatography (HPLC) with fluorescence detection (excitation at 265 nm and emission at 344 nm). The method employed one-step extraction of itopride from human plasma with tert-butyl methyl ether using alfuzosin hydrochloride as an internal standard. Chromatographic separation was obtained within 10 min using a reverse phase column (C18, 250 mm × 4.6 mm, 5 µm) and an isocratic mobile phase constituting of acetonitrile, tetrahydrofuran and pH 3.5 buffer (suitable ratio) at a flowrate of 1.2 ml/min. The method was linear in the range of 5 to 1000 ng/ml. The lower limit of quantitation was 5 ng/ml. The intra and inter-day accuracy were within between 86.2 % and 108.5 %. Plasma sample containing itopride were stable for 100 days at -35 ^oC and for 6 hours at room temperature. This method can be used for BA-BE studies of itopride preparations.