血浆华法林HPLC测定法及心脏换瓣术后患者血药浓度监测初探

Determination of Warfarin in Plasma by HPLC and an Investigation of M onitoring Patients after Cardiac Valve Replacement

目的 建立反相 HPL C法测定血浆中华法林浓度 ,并探讨心脏瓣膜替换术后患者华法林血药浓度与抗凝疗效的关系。方法 血浆样品经二氯甲烷 -正己烷 ( 1∶ 9)萃取浓集等预处理后 ,于国产 YWG C18柱上 ,以甲醇 - 5 0 mm ol/ L醋酸铵缓冲液 ( p H2 .5 ) ( 70∶ 30 )为流动相分离 ,紫外 30 8nm波长测定。结果 华法林最低检测浓度为 2 0 ng/ m l,标准曲线线性范围 5 0～ 2 0 0 0 ng/ m l。血浆华法林萃取回收率 78.0 %～ 81.6 % ,方法回收率10 3.1%～ 10 6 .5 % ,日内 RSD 2 .33%～ 5 .46 % ,日间 RSD 5 .2 9%～ 7.73%。用本法监测 70例心脏换瓣术后患者 ,结果 INR值在推荐国人安全范围的 44例中 ,37例 ( 84.1% )患者华法林血药浓度在 6 16 .2± 15 4.8ng/ m l。

Objective A simple HPLC method was established f or the determination of warfa rin in plasma to investigate the relationship between warfarin concentration a nd anticoagulant effect. Methods The mixture of dichloromethane and hexane (1∶9)was u sed as extracting solvent for the plasma samples. The chromatographic separat ion was on C 18 column with a mobile phase consisting of methanol and 50 m mol/ L ammonium acetate buffer (pH2.5,70∶30).Results The calibr ation curve was linear w ithin 50 2000 ng/ml. The extraction recoveries of warfarin were 78.0% 81.6%. Th e recoveries of methodology were 103.1% 106.5%. Inter day and intra day RSD w e re 2.33% 5.46% and 5.29% 7.73%, respectively. This method was used for determi ni ng warfarin in 70 patients after cardiac valve replacement. The results showed t hat 44 cases had their INR within the safety range(1.31 2.35) recommended to Ch inese, and of them 37(84.1%) cases had a warfarin level at 616.2±154.8ng/ml.Conclusion This method is useful in monitoring warfarin concentrat ion during anticoagulant therapy.