液相色谱-串联质谱法测定人血浆中伏立康唑和伏立康唑氮氧化物浓度及其临床应用

Determination of voriconazole and its N-oxide in human plasma by LC-MS/MS and its clinical application

目的建立同时测定人血浆中伏立康唑和伏立康唑氮氧化物浓度的液相色谱-串联质谱(LC-MS/MS)方法,并应用于临床伏立康唑的治疗监测。方法酮康唑为内标,经甲醇沉淀蛋白后进行LC-MS/MS分析。电喷雾离子源,正离子模式监测,检测离子伏立康唑m/z350.1→m/z127.0,伏立康唑氮氧化物m/z366.1→m/z224.0,酮康唑(内标)m/z531.0→m/z82.1;色谱柱为Inertsil ODS-3(2.1 mm×50 mm,5μm),流动相为甲醇∶10 mmol/L乙酸铵溶液(90∶10)。运用本法对本院23例患者进行伏立康唑和伏立康唑氮氧化物血药浓度测定。结果伏立康唑和伏立康唑氮氧化物在0.4~10.0μg/mL浓度范围内线性良好(r>0.999),最低定量限为0.4μg/mL。批内、批间精密度及基质效应RSD均小于15%。样本储存和处理过程中,化合物稳定良好。23例患者伏立康唑稳态谷浓度为0.4~14.3μg/mL,伏立康唑氮氧化物稳态谷浓度为0.5~7.2μg/mL,个体间差异较大。结论本文建立的液相色谱-串联质谱(LC-MS/MS)方法操作简便,特异性强,灵敏度高,可应用于临床治疗药物监测。

Objective To develop liquid chromatography-tandem mass spectrometry(LC-MS/MS)for the content determination of voriconazole(VRCZ)and its N-oxide(VNO)in human plasma,and to apply it to clinical samples.Methods Ketoconazole was used as the internal standard(IS).The plasma samples were treated with methanol to precipitate protein and analyzed by HPLC-MS/MS.The detection was accomplished with electrospray ionization(ESI)operated in positive ionization mode,and ion reaction pairs were selected to be m/z350.1→m/z127.0(VRCZ),m/z366.1→m/z224.0(VNO)and m/z531.0→m/z82.1(Ketoconazole).Chromatographic separation had been achieved on Inertsil ODS-3(50/2.1)with methanol-10 mmol/L ammonium acetate(90:10)as the mobile phase.Plasma concentrations of VRCZ and VNO in 23 patients were determined.Results Both VRCZ and VNO were linear(r>0.999)over the concentration ranges of 0.4~10.0μg/mL,inter-and intra-batch relative standard deviations(RSDs)and matrix effect were all less than 15%.The 2 compounds remained stable during sample storage and handling progress.Plasma concentrations of VRCZ in 23 patients ranged from 0.4 to 14.3μg/mL,and those of VNO ranged from 0.5 to 7.2μg/mL.Significant variance was found among these patients.Conclusion LC-MS/MS is simple,quick and sensitive for detection of VRCZ and VNO,and can be used for drug monitoring in clinical practice.