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目的建立心脏瓣膜置换术后患者血中华法林浓度的快速、准确的高效液相色谱法(HPLC),便于华法林临床药物浓度监测,实现个体化给药。方法收集西南医院、重庆医科大学附属第一医院人工机械瓣膜置换术后口服华法林抗凝治疗达稳态的患者100例的血浆样品,经正己烷-二氯甲烷(9∶1)萃取富集处理后,在310 nm紫外检测波长;流动相:甲醇∶水=75∶25(含0.2 ml冰乙酸);流速:0.9 ml/min;柱温:25℃条件下,经过Hypersil BDS C18(200 mm×4.6 mm,5μm)色谱柱分离测定。结果华法林的保留时间为6.41 min,样品浓度线性方程为C=3.12×10-3.R+0.038,r=0.999 8,线性范围为20～2 000 ng/ml,平均回收率为97.5%,日内与日间相对精密度均小于5%。对重庆地区100例心脏瓣膜置换术后患者血浆华法林测定,其血药浓度是(845.48±388.16)ng/ml。结论 HPLC法能快速、准确地测量心脏瓣膜置换术后患者血中华法林浓度,并能进行临床监测。
Objective To establish a rapid and accurate high performance liquid chromatography (HPLC) method for the determination of warfarin concentration in blood of patients after heart valve replacement, and to facilitate the monitoring of warfarin concentration in clinical medicine and achieve individualized administration. Methods Plasma samples from 100 patients with stable state of oral anticoagulant warfarin after oral mechanical valve replacement in Southwest Hospital and the First Affiliated Hospital of Chongqing Medical University were collected and extracted by n-hexane-dichloromethane (9: 1) After UV detection at 310 nm, the mobile phase consisted of methanol: water = 75:25 with 0.2 ml glacial acetic acid, flow rate: 0.9 ml / min, column temperature at 25 ° C and Hypersil BDS C18 4.6 mm, 5 μm) column. Results The retention time of warfarin was 6.41 min, the linear equation of sample concentration was C = 3.12 × 10-3.R + 0.038, r = 0.999 8, the linear range was 20-2 000 ng / ml and the average recovery was 97.5% , The relative precision of day and day were less than 5%. The plasma warfarin level was measured in 100 patients after heart valve replacement in Chongqing area. The plasma concentration was (845.48 ± 388.16) ng / ml. Conclusion HPLC method can quickly and accurately measure the concentration of warfarin in the blood of patients after heart valve replacement and can carry out clinical monitoring.