UPLC-MS/MS法检测人血浆中莫西沙星浓度

Determination of Moxifloxacin in Human Plasma Using Ultra Performance Liquid Chromatography Tandem Quadrupole Mass Spectrometry

关键词:  血药浓度  莫西沙星  药动学  UPLC-ESI-MS/MS

Key words:  plasma concentration  moxifloxacin  pharmacokinetics  UPLC-ESI-MS/MS

摘要:

目的 建立快速检测人血浆中莫西沙星浓度的UPLC-MS/MS方法。方法 用乙腈沉淀血浆蛋白的方法处理，运用Waters XEVO TQD三重四级杆液质联用仪，色谱柱为ACQUITY UPLC? BEH C₁₈柱(100 mm×2.1 mm，1.7 μm)，流动相为乙腈-水(含0.1%甲酸)，梯度洗脱，流速为0.4 mL·min^-1，柱温40 ℃，内标为左氧氟沙星，质谱条件：电喷雾离子化源(ESI)，正离子检测模式；加入200 μL冰乙腈沉淀蛋白，13 000 r·min^-1离心10 min，取200 μL上清于进样瓶内，取2 μL上样。结果 莫西沙星的保留时间为1.58 min，线性范围为0.1~10 μg·mL^-1(r=0.999 6)，最低定量限为0.01 μg·mL^-1，高、中、低浓度的相对回收率为(96.66±3.8)%，(102.0±1.76)%和(107.2±7.19)%。日内、日间RSD均<10%，血浆样品体系中的其他内源性物质不干扰测定。结论 该方法准确可靠，操作简便，重复性好，适于检测人血浆中莫西沙星的血药浓度及其药动学研究。

Abstract:

OBJECTIVE To construct an ultra performance liquid chromatography-tandem quadrupole mass spectrometry method to determine the concentration of moxifloxacin in human plasma. METHODS Precipitation method with acetonitrile was used. An ACQUITY UPLC? BEH C₁₈(100 mm×2.1 mm, 1.7 μm) column was used as the stationary phase at 40 ℃ on Waters XEVO TQD. The mobile phase was consisted of acetonitrile and water (containing 0.1% formic acid) with gradient elution pumped at a flow rate of 0.4 mL·min^-1. Plasmas were precipitated with 200 μL acetonitrile and centrifugated at 13 000 r·min^-1 for 10 min. RESULTS The retention time of moxifloxacin was 1.58 min. Excellent linear calibration curves of moxifloxacin was obtained in the concentration range of 0.1-10 μg·mL^-1(r=0.999 6). The lower limit of quantification of moxifloxacin was 0.01 μg·mL^-1, while the recovery of three concentrations were (96.66±3.8)%, (102.0±1.76)% and (107.2±7.19)%. The intra- and inter- day six standard deviations were all less than 10%. There were no endogenous substances existing in the incubation system which interfered with the determination of the analyses of interest. CONCLUSION The method is simple, accurate and rapid which is suitable for the determination of moxifloxacin and pharmacokinetics study in human.