论文部分内容阅读
目的:建立快速检测人血浆中华法林及其代谢产物7-羟基华法林浓度的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 UPLCBEH C18柱(50 mm×2.1mm,1.7μm);流动相为乙腈-水(含0.1%甲酸),梯度洗脱,流速为0.4 ml·min-1,柱温为40℃,内标为华法林-d5;质谱条件:电喷雾离子化源(ESI),正离子检测模式。结果:华法林和代谢产物7-羟基华法林的保留时间分别为1.8 min和1.5 min,线性范围分别为25~2 000 ng·ml-1(r=0.999 3)和5~500 ng·ml-1(r=0.999 6),最低定量限分别为5,2.5 ng·ml-1,回收率分别为96.9%~105.3%,97.1%~103.3%,日内、日间RSD均<10%。结论:该法准确可靠,操作简便,重复性好,适于检测人血浆中华法林及其代谢产物7-羟基华法林的浓度。
Objective: To establish a rapid UPLC-MS / MS method for the determination of warfarin and its metabolite 7-hydroxy warfarin in human plasma. Methods: The plasma protein was precipitated by acetonitrile. The sample was eluted with a Waters XEVO TQD Triple Quadrupole LC / MS column (50 mm × 2.1 mm, 1.7 μm). The mobile phase consisted of acetonitrile- Water (containing 0.1% formic acid) gradient elution at a flow rate of 0.4 ml · min-1 at a column temperature of 40 ° C with an internal standard of warfarin-d5; mass spectrometry conditions: electrospray ionization source Detection mode. Results: The retention times of warfarin and 7-hydroxy warfarin were 1.8 min and 1.5 min, respectively, with linear ranges of 25-2 000 ng · ml-1 (r = 0.999 3) and 5-500 ng · ml-1 (r = 0.999 6). The lowest limit of quantitation was 5 and 2.5 ng · ml-1, respectively. The recoveries were 96.9% -105.3% and 97.1% -103.3%, respectively. Conclusion: The method is accurate, reliable, simple and reproducible, and suitable for the determination of warfarin and its metabolite 7-hydroxy warfarin in human plasma.