【摘要】 目的:降低基质效应,建立测定人血浆中苯烯莫德的液相色谱-串联质谱(LC-MS/MS)法。方法:以氟代苯烯莫德为内标,血浆样品用四硼酸钠碱化后,用甲基叔丁基醚(MEBE)萃取,离心后取上清液氮气吹干,流动相复溶后进行LC-MS/MS分析。色谱条件:采用Ultra C18(50 mm×2.1 mm,5.0μm)色谱柱,以乙腈-0.2 mmol·L-1甲酸铵为流动相,流速300μL·min-1,柱温20℃;质谱条件:电喷雾离子化源(ESI),负离子模式多重反应选择离子检测(MRM),用于定量的离子对分别为苯烯莫德m/z 253.1→210.9和内标氟代苯烯莫德m/z 270.9→229.2。结果:人血浆中苯烯莫德的线性范围为0.1~10 ng·m L-1(r>0.99),定量下限为0.1 ng·m L-1;日内、日间精密度(RSD)均小于15%;相对误差为-1.03%~0.01%;提取回收率大于85%;稳定性良好,血浆基质对苯烯莫德测定无干扰。结论:该方法适用于人血浆中苯烯莫德浓度的测定。氟代同系物内标能部分取代稳定同位素内标,应用在生物样品的检测中。更多还原

【Abstract】 Objective: To reduce the matrix effect and establish a LC-MS / MS method for the determination of benvitimod in human plasma. Methods: Fluorophenyl-benvitimod was used as the internal standard. The samples were firstly alkalified with sodium tetraborate,and then extracted by methylterbutylethylene( MTBE). The supernatant liquid after centrifugation was dried by nitrogen and analyzed by the LC-MS / MS method. Analytes were separated on the Ultra C18column( 50 mm × 2. 1 mm,5. 0 μm) with the mobile phase consisting of acetonitrile and 0. 2mmol·L-1ammonium formate. The flow rate was 300 μL·min-1,and the column temperature was 20 ℃. Detection and quantitation was performed by electrospray ionization( ESI) and multiple reaction monitoring( MRM) in the negative ion mode. The ion pairs of m / z 253. 1 →210. 9 and m / z 270. 9 →229. 2 were used to qualify benvitimod and fluorophenyl-benvitimod( internal standard),respectively. Results: The calibration curve was linear over the concentration range of 0. 1-10 ng·m L-1( r > 0. 99),with the lower limit of quantitation( LLOQ) of 0. 1 ng·m L-1. Intra-day and inter-day precisions were less than 15% and the relative error was between-1. 03% and0. 01%. The extractive recovery was above 85%,stabilities were good. The plasma matrix showed no effect on the quantification of benvitimod. Conclusion: The established method can be applied in analysis of the benvitimod concentration in human plasma. Fluorinated homolog internal standard may partly replace stable isotopes as the internal standard in the application of biological samples.更多还原