衍生化β-环糊精液相色谱手性固定相拆分和测定人体血浆中卡维地洛对映体  

Eantioseparation and determination of carvedilol enantiomers in human plasma on a derivatized β-cyclodextrin-based chiral stationary phase by HPLC

作　　者：程彪平[1] 李来生[1,2] 周仁丹[2] 曹志刚[1] 曾春[1] 

机构地区：[1]南昌大学化学学院,江西南昌330031 [2]南昌大学分析测试中心,江西南昌330047

出　　处：《南昌大学学报（理科版）》2017年第1期77-82,89,共7页Journal of Nanchang University(Natural Science)

基　　金：国家自然科学基金资助项目(21165012);南昌大学食品科学与技术国家重点实验室开放基金(SKLF-KF-201605);江西省教育厅科技重点项目(GJJ150019)

摘　　要：以有序介孔SBA-15为基质,通过"点击化学"反应制备一种二硝基苯醚化β-环糊精键合SBA-15液相色谱手性固定相(NESP)。在极性有机溶剂模式下,成功地实现了β-受体阻滞剂卡维地洛对映体的快速手性分离,较详细地考察了流动相中甲醇含量、冰醋酸与三乙胺的浓度和温度等因素对手性分离的影响。在流动相组成为乙腈/甲醇/冰醋酸/三乙胺(91/9/1.0/1.0,v/v/v/v),流速为1.0mL·min^(-1),柱温为20℃,进样量为20μL,检测波长为285nm的色谱条件下,卡维地洛对映体在较短的时间(<15min)内达到基线分离,分离度为1.62。血液样品用叔丁基甲醚提取,以S-噻吗洛尔为内标,采用HPLC-MS选择离子[M+H]+407.30和317.20分别监测卡维地洛和内标S-噻吗洛尔,建立了一种测定血浆中卡维地洛对映体含量的新方法。卡维地洛两对映体在10~200ng·mL-1的浓度范围呈良好的线性关系(r=0.9992~0.9994),采用标准添加法测得两对映体在血浆中的回收率为88.60%~95.44%。血浆样品测定结果的日内和日间RSD分别≤1.73%和≤3.84%(n=5)。按三倍噪声推算出最小检测浓度为2.5ng·mL^(-1)。测定了健康人口服外消旋的卡维地洛后血浆中对映体的含量,发现卡维地洛的总含量和S-/R-对映体比例存在一定的差别,这与个体的差异和对映体生物利用度的不同有关。实验结果表明,所建立的分析方法灵敏、准确、简便、快速,自制的环糊精手性柱可大大降低测试成本,适用于人体血浆中卡维地洛对映体的测定及药代动力学研究。A novel of dinitrophenyl ether β-cyclodextrin-bonded SBA-15 chiral stationary phase（NESP） for HPLC was prepared by using ordered mesoporous matrix via a＂Click Chemistry＂reaction. The fast enanti- oseparation of carvedilol enantiomers on NESP was successfully achieved in ploar organ solvent mode. Some effect factors such as the methanol content, the ratio and concentration of glacial/triethylamine and temperature on the enantioresolution were investigated. The optimized composition of mobile phase was acetonitrile/methanol/glacial acetic acid/triethylamine（91/9/1/1,v/v/v/v） at a flow rate of 1.0 mL · min-1. The column temperature was at 20℃. The wavelength was at 285 nm. The injection volume was 20 μL. The carvedilol enantiomers were separated with resolution 1.62 within 15 min. The carvedilol was ex- tracted from human plasma with tert-butyl methyl ether. A new LC-MS method for the determination ofcarvedilol enantiomers in human plasma was established by selected positive ion monitoring of [M＋H]＋ 407.30 for the enantiomers and [M＋H]＋ 317.20 for S-timolol interal standard,respectively. The good lin- ear relationships for two carvedilol enantiomers were observed in the range of 10~200 ng · mL-1 （r=0. 9992~0. 9994）. The recoveries of two enantiomers from human plasma were 88.60%-95.44%. The RSDs were less than 1.73% for intra-day and 3.84% for inter-day（n=5）. The detection limits were 2.5 ng · mL-1 for both enantiomers. The total content of earvedilol and S-/R-enantiomeric ratio in healthy human plasma after oral administration racemic carvedilol existed certain difference, which related with individual differences and bioavailability of the enantiomers. The results showed that the established method was sim- ple, fast,sensitive,exact and low cost by using homemade cyclodextrin-based chiral column. It is suitable for fast quantification of carvedilol enantiomers and its pharmacokineties study.

关 键 词：液相色谱-质谱联用法 β-环糊精类SBA-15固定相 卡维地洛 对映体含量测定 人体血浆 

分 类 号：O657[理学—分析化学]