Hou Jingguo, Zheng Jie, Shamsi Shahab A
Department of Chemistry, Center of Biotechnology and Drug Design, Georgia State University, Atlanta, GA 30303, USA.
J Chromatogr A. 2007 Aug 3;1159(1-2):208-16. doi: 10.1016/j.chroma.2007.04.037. Epub 2007 Apr 21.
Warfarin is a widely used oral anticoagulant which is mostly administrated as a racemic mixture containing equal amount of R- and S-enantiomers. The two enantiomers are shown to exhibit significant differences in pharmacokinetics and pharmacodynamics. In this study, a new chiral micellar electrokinetic chromatography-mass spectrometry (MEKC-MS) method has been developed using a polymeric chiral surfactant, polysodium N-undecenoyl-L,L-leucyl-valinate (poly-L,L-SULV), as a pseudostationary phase for the chiral separation of (+/-)-warfarin (WAR) and (+/-)-coumachlor (COU, internal standard). Under optimum MEKC-MS conditions, the enantio-separation of both (+/-)-WAR and (+/-)-COU was achieved within 23 min. Calibration curves were linear (R=0.995 for (R)-WAR and R=0.989 for (S)-WAR) over the concentration range 0.25-5.0 microg/mL. The MS detection was found to be superior over the commonly used UV detection in terms of selectivity and sensitivity with LOD as low as 0.1 microg/mL in human plasma. The method was successfully applied to determine WAR enantiomeric ratio in patients' plasma undergoing warfarin therapy.
华法林是一种广泛使用的口服抗凝剂，大多以含有等量R-和S-对映体的外消旋混合物形式给药。已表明这两种对映体在药代动力学和药效学方面存在显著差异。在本研究中，开发了一种新的手性胶束电动色谱-质谱联用（MEKC-MS）方法，使用聚合手性表面活性剂N-十一碳烯酰-L,L-亮氨酰缬氨酸多钠盐（聚-L,L-SULV）作为假固定相，用于手性分离（±）-华法林（WAR）和（±）-氯灭鼠灵（COU，内标）。在最佳MEKC-MS条件下，（±）-WAR和（±）-COU的对映体分离在23分钟内完成。在0.25-5.0微克/毫升的浓度范围内，校准曲线呈线性（（R）-WAR的R = 0.995，（S）-WAR的R = 0.989）。发现MS检测在选择性和灵敏度方面优于常用的UV检测，在人血浆中的检测限低至0.1微克/毫升。该方法成功应用于测定接受华法林治疗患者血浆中的WAR对映体比例。
