HPLC-MS/MS法同时测定人血浆中伏立康唑及其氮氧化代谢物浓度

An LC-MS/MS Method for the Simultaneous Determination of Voriconazole and Voriconazole N-oxide in Human Plasma and for the Pharmacokinetic Study

目的:建立快速测定人血浆中伏立康唑及其主要代谢物伏立康唑-N-氮氧化物浓度的HPLC-MS/MS方法,用于人体药代动力学研究。方法:以伏立康唑和伏立康唑-N-氮氧化物的D3同位素为内标,血浆样品经蛋白沉淀后,用Agilent ZORBAX SB-Aq色谱柱(2.1 mm×50 mm,1.8μm)和API 4000质谱仪进行正离子模式电喷雾离子化分析。流速为0.6 mL·min^-1,以甲醇为有机相,0.1%甲酸为水相,按一定程序进行梯度洗脱。结果:伏立康唑和伏立康唑-N-氮氧化物在10.00~8000 ng·mL^-1范围内线性关系良好,该方法的特异性、批内/批间精密度、准确度、介质效应、提取回收率、稳定性和稀释可靠性符合相关要求。结论:建立了一种准确、简便、可靠的HPLC-MS/MS分析方法,成功应用于健康中国人静脉注射4 mg·kg^-1伏立康唑后的药代动力学研究。

Objective:To establish an HPLC-MS/MS method for the rapid determination of voriconazole and its main metabolite voriconazole N-oxide in human plasma for the study of voriconazole pharmacokinetics.Methods:With voriconazole-D3 and voriconazole-D3 N-oxide as isotopic internal standards,plasma samples were processed by protein precipitation and analyzed by an Agilent ZORBAX SB-Aq column(2.1 mm×50 mm,1.8μm)and an API 4000 MS with positive electro spray ionization.The flow rate was set at 0.6 mL·min^-1.Gradient elution was carried out with the ratio of methanol and 0.1%formic acid varied as scheduled.Results:The calibration curves for both voriconazole and voriconazole N-oxide ranged from 10.0 to 8000 ng·mL^-1 in human plasma.The specificity,intra-/inter-run precisions,accuracy,matrix effects,extraction recoveries,stabilities as well as the dilution integrity of this method met the relevant requirements.The method was verified with two healthy Chinese subjects under single intravenous injection of 4 mg·kg^-1 voriconazole.Conclusions:An accurate,simple and reliable HPLC-MS/MS method has been developed.The developed method can be used for the pharmacokinetic study of voriconazole.