液相色谱-串联质谱法测定人血浆中阿普唑仑的浓度

Determination of alprazolam in human plasma by liquid chromatography-tandem mass spectrometry

目的建立一种测定人血浆中阿普唑仑浓度的液相色谱-串联质谱(liquid chromatography-tandem mass spectrometry,LC-MS/MS)法。方法用蛋白沉淀法对样品进行前处理。色谱柱:Inert Sustain AQ-C_(18) HP(2.1 mm×50.0 mm,3μm),流动相:100%水含2 mmol·L^(-1)醋酸铵和0.1%甲酸-100%甲醇含0.1%甲酸,流速:0.60 mL·min^(-1),柱温:40℃,进样量:10μL。用电喷雾离子化源,正离子模式,多反应监测。考察该方法的专属性、标准曲线和定量下限、精密度与回收率、稀释效应、稳定性。结果阿普唑仑在0.10~15.00 ng·mL^(-1)线性关系良好,其标准曲线为y=2.14×10^(-1)x+9.41×10^(-4)(r=0.9986),定量下限为0.10 ng·mL^(-1),日内和日间相对标准偏差为1.4%~6.6%,稀释效应的相对标准偏差为4.0%,回收率可达93%以上。结论本方法的专属性强、灵敏度高,适用于阿普唑仑的治疗药物监测和药代动力学研究。

Objective To establish a liquid chromatography-tandem mass spectrometry(LC-MS/MS)method for the determination of alprazolam in human plasma.Methods Samples were processed by the protein precipitation method.The chromatographic column was Inert Sustain AQ-C_(18) HP(2.1 mm×50.0 mm,3μm).Mobile phase was 100% water contains 2 mmol·L^(-1) ammonium acetate and 0.1% formic acid-95% acetonitrile contains 2 mmol·L^(-1) ammonium acetate and 0.1% formic acid.Flow rate was 0.60 mL·min^(-1).The column temperature was 40℃.The injection volume was 10μL.The specificity,standard curve and lower limit of quantification,precision and recovery,dilution effect and stability were investigated by electric spray ionization source in positive ion mode with multiple reaction monitoring.Results Alprazolam was well related linearly within 0.10-15.00 ng·mL^(-1),and the standard curve was y=2.14×10^(-1)x+9.41×10^(-4)(r=0.9986),the lower limit of quantification was 0.10 ng·mL^(-1),the relative standard deviations of intra-day and inter-day were 1.4%-6.6%,the relative standard deviations of dilution effect was 4.0%,and the recovery rate was more than 93%.Conclusion This method has strong specificity and sensitivity and is suitable for pharmacokinetic studies and therapeutic drug monitoring of alprazolam.