LC-MS/MS法测定人血浆中米卡芬净浓度

Determination of micafungin in human plasma by LC-MS/MS

目的建立测定人血浆中米卡芬净的液相色谱-串联质谱(LC-MS/MS)法,验证后应用于米卡芬净在侵袭性真菌感染患者中的药代动力学研究。方法血浆样品用乙腈(含0. 1%甲酸)沉淀蛋白预处理,内标选用西替利嗪,经Agilent poro-shell 120SB-C_(18)(2. 1 mm×100 mm,2. 7μm)色谱柱进行分离,流动相为乙腈^(-1)0 mmol·L^(-1)醋酸铵溶液(均含0. 1%甲酸)梯度洗脱,流速为0. 20m L·min^(-1),用电喷雾离子化源,正离子方式,扫描方式为多反应监测(MRM),用于监测的离子反应分别为m/z 1271. 0→m/z 1172. 3(米卡芬净)和m/z 398. 1→m/z 212. 0(内标西替利嗪)。方法学验证后用于患者血浆中米卡芬净浓度测定。结果血浆中内源性物质对测定无干扰,米卡芬净在0. 10～25μg·m L^(-1)线性关系良好(r=0. 998 1); 4个质控浓度水平(定量下限,低、中、高质量浓度)的准确度在93. 8%～107. 9%;批内和批间精密度良好,RSD均小于15%;米卡芬净和内标的提取回收率在83. 4%～93. 3%,内标归一化基质效应为93. 9%～104. 8%。静脉滴注米卡芬净,每次100 mg,每日1次,连续用药7 d后,Cmax为(8. 87±1. 95)μg·m L^(-1),t_(1/2)为(16. 85±2. 23) h,血药浓度-时间曲线下面积(AUC_(0-t))为(98. 94±18. 57)μg·m L^(-1)·h,AUC_(0-∞)为(107. 41±21. 30)μg·m L^(-1)·h,表观分布容积(Vz)为(29. 52±7. 49) L·kg^(-1)。结论本方法简单、快速、灵敏、专属性好可用于人血浆中米卡芬净浓度的测定并用于药代动力学研究。

Objective To develop a LC - MS/MS method for the determination of micafungin in human plasma and apply this method of phar- macokinetic characteristics in patients with invasive fungal infections Methods The analyte were extracted from plasma by protein precipita- tion with acetonitrile （containing 0. 1% formic acid）, cetirizine was selected as internal standard. Chromatographic separation was achieved on Agilent poro - shell 120SB - C18 Column （2. 1 mm×100 mm, 2.7μm）, with gradient elution （ A ： acetonitrile, B ： 10 mmol·L^-1 ammonium acetate ） at the flow rate of 0. 20 mL·min^-1 Quantification was performed with the positive multiple reaction monitoring （MRM）. The MS/MS ion transitions monitored were m/z 1271.0→m/z 1172.3 and m/z 465.3→m/z 270. 1 for micafungin and internal standard, respectively. After methodology validation, this method was applied for the clinical analysis of micafungin in plasma from patients. Results Chromatograms showed no endogenous interfering peaks with blank samples. The standard curves were linear in the range of 0. 10 - 25 μg·mL^-1 （r =0. 9981 ）. The RSD of inter-day and intra- day for four different concentration levels were less than 15%, the average extraction recoveries were between 83.4% -93.3%. The internal standard normalized matrix effect was around 93.9% -104. 8%. After the intravenous infusion of micafungin 100 mg, qd for consecutive 7 d, the main pharmacoki- netic parameters were as follows Cmx： （8. 87 ± 1.95） μg·mL^-1, t1/2： （ 16. 85±2.23） h, AUC0-1 （98.94±18.57） μg. mL^-1·h, AUC0-∞ （107.41 ±21.30） μg·mL^-1·h, Vz （29.52 ±7.49） L·kg^-1 Conclusion The established method is simple, rapid, specific, sensitive and can be used to determine the concentration of micafungin in human plasma.