摘要
目的:建立HPLC-MS/MS法测定深静脉血栓患者血浆中华法林对映异构体的浓度。方法:以华法林-d5为内标,经甲醇沉淀蛋白后,采用CHIRALPAK?AS-3R手性色谱柱(4.6 mm×150 mm,3μm)分离样品,流动相为甲醇-含0.1%甲酸的醋酸铵水溶液(95∶5)。采用质谱检测系统,电喷雾离子源(ESI源),负离子扫描,多反应监测模式(MRM)监测m/z 307.0→160.8(华法林)及m/z 313.0→160.8(华法林-d5)。采集77名深静脉血栓患者稳态血样后,以HPLC-MS/MS法测定血浆中华法林对映异构体的浓度,并计算S-华法林与R-华法林浓度的比值(CS/R)。结果:华法林的2个对映异构体质量浓度在5~1 500 ng·mL-1内线性关系良好,定量下限为5 ng·mL-1。R-华法林批内和批间精密度的RSD为5.0%~8.1%和8.3%~13.4%,提取回收率85.9%~97.7%;S-华法林批内和批间精密度的RSD为6.2%~7.0%和6.8%~15.0%,提取回收率82.4%~93.9%。2个异构体均无基质效应影响,不同条件下的稳定性试验均满足要求。77名患者体内R-华法林与S-华法林稳态血浆药物质量浓度分别为(458.52±226.55)ng·mL-1和(239.63±152.83)ng·mL-1,其血浆药物浓度比值的平均值约为0.53±0.34。结论:本方法快速、简便、可靠,适用于深静脉血栓患者体内华法林对映异构体血药浓度的检测,能够指导临床合理用药。
Objective:To establish a simple,rapid and reliable HPLC-MS/MS method for the determination of the concentration of both warfarin enantiomers in plasmas of patients with deep venous thrombosis. Methods:Warfarin-d5 used as the internal standard,after precipitating by methanol,the seperation of plasma was accomplished by a CHIRALPAK?AS-3 R(4.6 mm×150 mm,3 μm) chiral column with a mobile phase comprising of methanol-ammonium acetate sulution containing 0.1% formic acid(95∶5)at a flow rate of 0.6 mL·min-1,and then the detection was performed on the MS system. Detection and quantitation was done in multiple reaction mode(MRM) following the transitions m/z 307.0 → 160.8 and 307.0 → 160.8 for warfarin enantiomers and the warfarin-d5 diastereomers,respectively,in the negative ionization mode. After the steadystate blood samples of 77 patients with deep vein thrombosis were collected,the concentration of warfarin enantiomers in plasma was determined by HPLC-MS/MS method,and the concentration ratio of S-warfarin to R-warfarin(CS/R) was calculated. Results:The linearity range was 5-1 500 ng·mL-1 and the lower limit of quantification was 5 ng·mL-1 for each enantiomers. The intra-and interbatch precisions were in the ranges of 4.99%-8.13% and 8.27%-13.41%,and 6.21%-6.99% and 6.82%-14.99% for R-warfarin and S-warfarin,respectively. The mean extraction recovery rate of both analytes ranged from 82.4% to 97.7%,the effect of the biological matrix was negligible. Both enantiomers were found to be stable under different stability conditions. In 77 patients,the steady plasma concentrations of R-warfarin and S-warfarin were(458.52±226.55) ng·mL-1 and(239.63±152.83) ng·mL-1,respectively,with an average CS/R ratio of about 0.53±0.34. Conclusion:The established HPLC-MS/MS method is rapid,simple and reliable,and is suitable for detecting the plasma concentration of warfarin enantiomers in patients with deep vein thrombosis,and can guide the clinical rational use of drugs.
作者
梁大虎
徐毛迪
吴子静
何春远
袁小龙
孙华
李相鸿
谢海棠
LIANG Da-hu;XU Mao-di;WU Zi-jing;HE Chun-yuan;YUAN Xiao-long;SUN Hua;LI Xiang-hong;XIE Hai-tang(Anhui Provincial Center for Drug Clinical Evaluation,Yijishan Hospital of Wannan Medical College,Wuhu 241000,China;Wannan Medical College,Wuhu 241002,China;Department of Clinical Pharmacy,Lu’an People’s Hospital,Lu’an 237000,China;Department of Pharmacy,the Second Affiliated Hospital of Wannan Medical College,Wuhu 241000,China)
出处
《药物分析杂志》
CAS
CSCD
北大核心
2020年第2期232-239,共8页
Chinese Journal of Pharmaceutical Analysis
基金
安徽省教育厅高校自然科学基金项目(KJ2017A248)。
关键词
华法林
异构体
人血药浓度
液质联用法
深静脉血栓
warfarin
enantiomers
human plasma drug concentration
HPLC-MS/MS
deep vein thrombosis
