HPLC-MS/ESI同时测定人血浆中氯唑沙宗及其代谢物的浓度

 目的建立高效液相色谱质谱联用测定人血浆中氯唑沙宗及其代谢物6-羟基氯唑沙宗的方法。方法采用Thermo C₁₈色谱柱(4.6 mm×150 mm,5μm),Phenomenex C₁₈保护柱,柱温40℃,流动相:乙腈-30 mmol·L^-1醋酸铵溶液(34∶66),流速:1mL·min^-1,进入MS检测器的分流比3∶1。质谱采用电喷雾电离源负离子模式(ESI-),选择性监测质荷比(m/z)168.3,170.2(氯唑沙宗),444.7(内标),184.4,186.2(6-羟基氯唑沙宗)的准分子离子峰。血浆样品孵化后,经叔丁基甲醚提取,内标法定量。结果氯唑沙宗及6-羟基氯唑沙宗分别在25～10 000,10～3 000μg·L^-1内线性关系良好(r≥0.999 8);最低检测质量浓度分别为2.5和2μg·L^-1;两个化合物的方法回收率在90%～110%之间;日间和日内RSD均小于15%。结论该方法灵敏度高,快速,简便,可用于氯唑沙宗药动学及药物相互作用的研究。

Determination of Chlorzoxazone and Its Metabolite in Human Plasma by HPLC-MS

OBJECTIVE To establish a high performance liquid chromatography-electrospray mass spectrometry method for the determination of chlorzoxazone and its metabolite in human plasma.METHODS The HPLC separation was performed on a Thermo C18 column(4.6 mm×150 mm,5 μm)with guard column phenomenex C18.The column temperature was 40 ℃.The mobile phase consisted of acetonitrile-30 mmol·mL^-1 ammoniumacetate solution.The flow rate was 1 mL·min^-1 with a split ratio 3∶1.The compound was ionized in the electrospray ionization(ESI) negative ion source of mass spectrometer and selected ions were at m/z 168.3,170.2(CZX),444.7(IS)and 184.4,186.2(HCZX).Human plasma samples were extracted with methyl t-butyl ether after incubation.RESULTS The linear ranges were 25～10 000 μg·L^-1 for chlorzoxazone,10～3 000 μg·L^-1 for 6-hydroxychlorzoxazone(r≥0.999 8),and their detect limits were 2.5,2.0 μg·L^-1.Recoveries were within 90%～110%,and intra-and inter-day RSDs were all less than 15%.CONCLUSION The methed appeared to be accurate,sensitive,specific and simple.It is suitable for clinical study of chlorzoxazone pharmacokinetics and drug interaction.