RP-HPLC法测定人血中伏立康唑血药浓度

Determination of Plasma Concentrations of Voriconazole in Human Blood by RP-HPLC

目的建立一种快速、简便、准确的RP-HPLC法测定人血中伏立康唑血药浓度的方法,以便于监测伏立康唑的血药浓度,指导患者个体化治疗。方法采用2倍体积乙腈沉淀血浆蛋白,采用RP-HPLC法,以卡马西平为内标,色谱条件如下:色谱柱为Agilent TC-C_(18)(4.6 mm×250 mm,5μm),流动相为乙腈-水(40∶60,V/V),流速1.0 ml·min^(-1),紫外检测波长为255 nm,进样量10μl。结果伏立康唑与卡马西平的保留时间分别为10.1和6.52 min。伏立康唑在0.2256~22.56μg·ml^(-1)范围内线性关系良好(r=0.9998);定量下限为0.2256μg·ml^(-1);相对标准偏差分别批内≤4.30%,批间≤3.94%;伏立康唑高、中、低三个浓度的绝对回收率为80.51%~95.44%。结论该方法灵敏度高,操作简便,结果准确,检测成本低,更适用于临床实践中常规的伏立康唑血药浓度监测及基础药代动力学研究。

Objective To establish a rapid,simple and accurate method for the determination of Voriconazole in human blood by Reversed-Phase High-Performance Liquid Chromatography(RP-HPLC)in order to facilitate the monitoring of Voriconazole concentrations and individualized treatment of patients.Methods The plasma protein was precipitated using the 2-fold volume acetonitrile method.RP-HPLC was used with carbamazepine as the internal standard.The chromatographic column was an Agilent TC-C18 column(4.6 mm×250 mm,5μm),with acetonitrile-water(40∶60,V/V)as the mobile phase.The flow rate was 1.0 ml·min^(-1)and the detection wavelength was 255 nm.The sample size was 10μl.Results The retention time of Voriconazole and carbamazepine was 10.1 and 6.52 min,respectively.Voriconazole had a good linear relationship over the range of 0.2256-22.56μg·ml^(-1)(r=0.9998),and the lower limit of quantitation was 0.2256μg·ml^(-1).The results of the relative standard deviation of intraand inter-assay precision were less than 4.30%and 3.94%,respectively.The absolute recovery of Voriconazole ranged from 80.51%to 95.44%.Conclusion This method is highly sensitive,user-friendly,accurate and cost-effective,which is more suitable for monitoring of routine Voriconazole concentrations in clinical practice and basic pharmacokinetic studies.