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建立大鼠肝微粒体中盐酸度洛西汀的RP-HPLC测定方法,为其体外代谢研究奠定基础。盐酸度洛西汀与大鼠肝微粒体共孵育后,用甲醇沉淀蛋白。离心上清液,采用HP1100色谱系统,以甲醇-0.025mol.L-1磷酸二氢钾缓冲盐(62∶38,V/V,pH 3.5)为流动相,流速0.9mL.min-1,进样量20μL,经Hedera ODS-2(4.6mm×250mm,5μm)色谱柱分离,检测波长290nm。盐酸度洛西汀在0.4—16μg.mL-1范围内线性关系良好(r=0.9992),检测限为40ng.mL-1,定量限为0.2μg.mL-1,方法回收率在94.4%—105.0%之间,日内、日间精密度分别<5%和<10%(n=5)。本方法较为快速,灵敏,准确,可用于盐酸度洛西汀在大鼠肝微粒体中的体外代谢研究。
To establish an RP-HPLC method for the determination of duloxetine hydrochloride in rat liver microsomes, which lays a foundation for its in vitro metabolism research. After duloxetine hydrochloride is incubated with rat liver microsomes, the protein is precipitated with methanol. The supernatant was centrifuged and analyzed by HP1100 chromatography system with methanol-0.025mol.L-1 potassium dihydrogen phosphate buffer (62:38, V / V, pH 3.5) as mobile phase at a flow rate of 0.9mL.min-1 The sample volume was 20 μL and separated on a Hedera ODS-2 (4.6 mm × 250 mm, 5 μm) column with a detection wavelength of 290 nm. The duloxetine hydrochloride showed good linearity (r = 0.9992) in the range of 0.4-16μg.mL-1, the detection limit was 40ng.mL-1, the limit of quantification was 0.2μg.mL-1, the recovery was 94.4% 105.0%, intra-day and inter-day precision <5% and <10% respectively (n = 5). The method is rapid, sensitive and accurate and can be used to study the in vitro metabolism of duloxetine hydrochloride in rat liver microsomes.