用HPLC-MS/MS法考察两种伏立康唑注射剂在大鼠体内的药动学及组织分布情况

Determination of pharmacokinetics and drug distribution in the tissues of two kinds of voriconazole injections in rats by HPLC-MS/MS method

目的:建立测定Wistar大鼠血浆中伏立康唑浓度的高效液相色谱-串联质谱法(HPLC-MS/MS),考察两种伏立康唑注射剂及其代谢物伏立康唑氮氧化物在大鼠体内的药动学及组织分布。方法:把大鼠随机分为两组,从尾静脉分别注射国产和进口的伏立康唑注射剂,于不同时间点取血或脏器,采用HPLC-MS/MS法测定其浓度,流动相为乙腈-0.2%乙酸水溶液(内含20mmol/L乙酸铵)=80∶20(V/V),流速为0.5ml/min,以氟康唑为内标,采用多反应监测(MRM)方式检测伏立康唑和伏立康唑氮氧化物。检测离子对:伏立康唑m/z350.2→281.0,伏立康唑氮氧化物m/z366.1→224.0,内标氟康唑m/z307.1→220.1。采用PhoenixWinNolin64数据处理软件的非房室模型,以统计矩法比较伏立康唑及代谢物伏立康唑氮氧化物在心脏、肝脏、脑组织、肺、脾脏、肾脏中的组织分布。结果:伏立康唑及其氮氧化物的浓度与峰面积线性良好,精密度、重复性、稳定性良好。国产和进口伏立康唑注射剂中伏立康唑的血浆AUC0~24h分别为(244.4±25.1)、(207.1±19.1)μg·h·ml^-1;伏立康唑氮氧化物的血浆AUC0~24h分别为(60.5±8.6)、(61.3±5.8)μg·h·ml^-1。国产和进口制剂中伏产康唑和伏立康唑氮氧化物在各组织的AUC0~24h无显著性差异。结论:该方法简单快速,灵敏度高,可用于伏立康唑的体内过程研究。大鼠单剂量静脉注射国产和进口伏立康唑注射剂后,伏立康唑和伏立康唑氮氧化物在大鼠体内的主要药动学参数无显著性差异,在各组织的总体暴露未见显著性差异。

Objective:To establish a method for the determination of voriconazole in plasma of Wistar rats by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS),and also to study the pharmacokinetics and distribution of the metabolites voriconazole nitrogen oxides in the tissues of 2 kinds of voriconazole injections in rats.Methods:The rats were randomly divided into 2 groups.Domestic and imported voriconazole injections were respectively injected in tail veins,and then blood and tissue samples were collected at different time points.The drug concentrations were detected by HPLC-MS/MS method.The mobile phase was acetonitrile-0.2% glacial acetic acid aqueous solution (containing 20 mmol/L ammonium acetate) 80∶20( V/V ),and flow rate was 0.5 ml/min.The multiple reaction monitoring (MRM) was used to detect the levels of voriconazole and voriconazole nitrogen oxides.The detected ion pairs were m/z 350.2→281.0 (voriconazole),m/z 366.1→224.0 (vorconazole nitrogen oxides) and m/z 307.1→ 220.1 (fluconazole). The pharmacokinetic parameters of the 2 injections were determined by non- compartment model (statistical moment method) using Phoenix WinNolin 64 data processing software.The distributions of voriconazole and voriconazole nitrogen oxides in the tissues of the heart,liver,brain,lung,spleen and kidney were analyzed statistically.Results:Voriconazole and voriconazole nitrogen oxides were in good linearity with peak area.The precision,reproducibility and stability were good. AUC 0-24 h of voriconazole for the domestic and imported voriconazole injections was (244.4± 25.1) and (207.1±19.1)μg·h·ml^-1. AUC 0-24 of Voriconazole nitrogen oxides were respectively (60.5± 8.6) and (61.3 ± 5.8)μg·h·ml^-1. Conclusion:This method is simple,rapid and sensitive,and could be used to study in vivo pharmacokinetics of voriconazole.After giving single dose of domestic and imported voriconazole injections in the veins of the rats,no significant differences could be seen in the main pharmacokinetic parameters and overall exposure in the tissues.