应用抗生物素蛋白-脂质体复合物为手性选择剂的毛细管电色谱法拆分手性对映体

应用抗生物素蛋白-脂质体复合物为手性选择剂的毛细管电色谱法拆分手性对映体

依明·尕哈甫, 陈鑫, 郑丽英, 殷其蕾, 贺锐锐, 詹先王, 王耀欣, 何希辉, 韩南银^*

北京大学医学部药学院化学生物学系, 北京 100191

收稿日期:2010-03-12 修回日期:2010-06-10 出版日期:2010-07-15 发布日期:2010-07-15
通讯作者: 韩南银*

Enantiomers separation using avidin-liposome complex as a chiral selector in capillary electrophoresis

Yiming·Gahafu, Xin Chen, Li-Ying Zheng, Qi-Lei Yin, Rui-Rui He, Xian-Wang Zhan, Yao-Xin Wang, Xi-Hui He, Nan-Yin Han^*

Department of Chemical Biology, School of Pharmaceutical Sciences, Peking University Health Science Center, Beijing 100191, China

Received:2010-03-12 Revised:2010-06-10 Online:2010-07-15 Published:2010-07-15
Contact: Nan-Yin Han*

摘要/Abstract

摘要：

抗生物素白蛋白(Avidin)-脂质体复合物是一种特殊的手性选择剂, 本研究利用毛细管电色谱 (CEC) 将它应用于拆分D,L-色氨酸、D,L-苯乙内酰硫脲丝氨酸、D,L-苯乙内酰硫脲苏氨酸和R,S-盐酸吡格列酮对映体。Avidin 被固定在涂布于毛细管内的脂质体上, 该脂质体由二棕榈酰磷脂酰乙醇胺 (Biotinyl-cap-DPPE) 和不同比例的磷酯酰丝氨酸 (PS) 构成。实验结果发现将1 mg/mL 的Avidin和 Biotinyl-cap-DPPE/PS (80:20, mol%) 的脂质体作为毛细管固定相, 可以实现较好的分离度和重现性。本实验还研究了不同的涂布和预处理方法对手性拆分结果的影响, 最后, 还通过成功地进行了盐酸吡格列酮对映体手性拆分对该电色谱法进行了评估。因此, 利用制备毛细管固定相应用于手性对映体拆分的毛细管电色谱法在将来的药物开发研究中具有重要的现实意义。

关键词: Biotinyl-cap-DPPE, 抗生物素蛋白, 脂质体, 对映体拆分, 毛细管电色谱, 盐酸吡格列酮

Abstract:

Avidin-liposome complex is a specific chiral selector used to separate the enantiomers of D,L-tryptophan, D,L-phenylthiohydantoin-serine (D,L-PTH-Ser), D,L-phenylthiohydantoin-threonine (D,L-PTH-Thr) and R,S-pioglitazone hydrochloride in capillary electrochromatography (CEC). The avidin is immobilized on the phospholipids coated in the capillary. The liposome used for the phospholipid coating contains 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(Cap biotinyl) (sodium salt) (Biotinyl-cap-DPPE) (16:0) and different proportions of L-α-phosphatidylserine (PS) in Tris (hydroxymethyl) aminomethane (Tris) buffer at pH 7.4. A good resolution and reproducibility was obtained by coating the capillary with Biotinyl-cap-DPPE/PS (80:20, mol%) followed by immobilization of 1 mg/mL of avidin solution in N-(2-hydroxyethyl) piperazine-N'-(2-ethanesulfonic acid) (HEPES) buffer at pH 7.4. A comparative study of chiral separation efficiency with different capillary coating methods and preconditioning conditions was conducted. Finally, the electrochromatographic method was successfully used to separate enantiomers of pioglitazone hydrochloride. Therefore, coated CEC will be a promising tool for pharmaceutical enantiomers separation in new drug development.

Key words: Biotinyl-cap-DPPE, Avidin, Liposome, Enantiomers separation, CEC, Pioglitazone hydrochloride

中图分类号:

R917

Supporting: ^*Corresponding author. Tel.: 86-10-82801590; fax: 86-10-82802414

依明·尕哈甫, 陈鑫, 郑丽英, 殷其蕾, 贺锐锐, 詹先王, 王耀欣, 何希辉, 韩南银^*

应用抗生物素蛋白-脂质体复合物为手性选择剂的毛细管电色谱法拆分手性对映体

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Yiming·Gahafu, Xin Chen, Li-Ying Zheng, Qi-Lei Yin, Rui-Rui He, Xian-Wang Zhan, Yao-Xin Wang, Xi-Hui He, Nan-Yin Han^*

. Enantiomers separation using avidin-liposome complex as a chiral selector in capillary electrophoresis [J]. .

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