反相高效液相色谱法测定人血浆中罗红霉素的浓度

Determination the Concentration of Roxithromycin in Human Plasma by RP-HPLC

目的:建立测定人血浆中罗红霉素浓度的反相高效液相色谱方法。方法:采用Agilent C_(18)分析柱(150 mm×4.6 mm,3.6μm),以甲醇-0.02 mol·L^(-1)乙酸铵(68:32,用氨水调pH至7.4)为流动相,流速0.8 ml·min^(-1),紫外检测波长210 nm,柱温55℃,血浆样品碱化后用乙醚提取2次,以克拉霉素为内标。结果:罗红霉素的保留时间为6.2 min,线性范围为0.25～33μg·ml^(-1)(r=0.999 7),最低检测限4 ng,方法回收率在96.2%～100.1%之间,日内、日间RSD均小于4%。结论:该法简便快速、准确灵敏、重现性好,适用于罗红霉素血药浓度测定。

Objective ： To establish a method for determination the concentration of roxithromycin in human plasma by RP-HPLC. Method： The Agilent C18 column （150 mm×4.6 mm, 3.6μm） was used. The mobile phase consisted of methanol-0. 02 mol·L^-1 ammonium acetate（68： 32, adjusted to pH 7.4 with ammonia water） and the flow rate was 0.8 ml · min^-1. The detection wavelength was 210 nm. Roxithromycin and clarithromycin （internal standard） were extracted from alkalinized plasma （0.5 ml） with ether. Resuit： The retention time for roxithromycin and clarithromycin were 6.2 min and 5.2 min, respectively. The standard curve was linear in the concentration range of 0.25 - 33 μg·ml^-1, r = 0. 999 7. The recovery of roxithromycin was within 96.2% - 100.1%. The RSD of within-day and between-day were less than 4%. Conclusion： The method is simple, sensitive, and reproducible for the determination of roxithromycin concentration in human plasma.