摘要
目的建立液相色谱串联质谱(LC-MS/MS)法测定人血浆中达比加群浓度。方法以达比加群自身标准溶液(6.24 ng·mL^(-1))为内标,色谱柱为Aston SN苯基柱(2.1 mm×50 mm,5 μm),流动相采用甲醇-水溶液(含甲酸和氨水浓度分别为4 mmol·L^(-1))(33∶67,V/V)进行等度洗脱。流速0.8 mL·min^(-1),柱温45℃,进样量1 μL。基于全自动二维液相色谱串联质谱耦合仪和本研究组开发的阀切换技术,待测样品和内标依次进样,进样时间间隔0.4 min。在电喷雾离子化源(ESI)正离子条件和多反应监测模式(MRM)下扫描,检测离子对为m/z 472.15/289.10。结果达比加群在5.204~520.4 ng·mL^(-1)与峰面积线性关系良好(r=0.9997),日内、日间精密度<3%,准确度为91.75%~113.16%,提取回收率为88.03%~92.42%,基质效应为100.35%~102.84%。结论本方法首次以达比加群自身为内标,方法新颖,样品处理简单,分析过程快速、高效,测定结果准确,适用于达比加群血药浓度的测定。
Objective To establish an LC-MS/MS method to determine the concentration of dabigatran in human plasma.Methods The standard solution of dabigatran was used as the internal standard.The separation was conducted on an Aston SN phenyl column (2.1 mm×50 mm,5μm).The mobile phase was composed of methanol-deionized water (containing 4 mmol·L^(-1) of formic acid and ammonia)at 33∶67 (v/v).The flow rate was 0.8 mL·min^(-1).The column temperature was 45℃.The injection volume was 1μL.With the fully automatic two-dimensional liquid chromatography tandem mass spectrometer coupling instrument and the valve switching program developed,the sample and IS were injected respectively at an interval of 0.4 min.Electrospray ionization source under positive ion and multiple reaction monitoring mode were used.Ion transition was monitored with mass-to-charge ratio of m/z 472.15/289.10 for the quantification.Results This method showed good linearity at the concentration of 5.204-520.4 ng·m L;(r=0.9997).The intra-and inter-day precisions were less than 3%and the accuracy was at 91.75%-113.16%.The mean extraction recovery was 88.03%-92.42%.The mean matrix effect was 100.35%-102.84%.Conclusion This novel method uses dabigatran as the IS,simplifies the sample preparation,and analyzes rapidly and accurately,which is suitable for determining the concentration of dabigatran in human plasma.
作者
王峰
何昕
钱懿轶
WANG Feng;HE Xin;QIAN Yi-yi(Department of Pharmacy,the Second Xiangya Hospital of Central South University,Changsha 410011;Department of Pharmacy,Fuwai Yunnan Cardiovascular Hospital,Kunming 650000;Graduate School of Kunming Medical University,Kunming 650500)
出处
《中南药学》
CAS
2021年第12期2520-2524,共5页
Central South Pharmacy
基金
云南省教育厅科学研究基金项目(No.2018JS245)
云南省阜外心血管病医院院级科研基金项目(No.2019YFKT-08)。
