化学学报 ›› 2014, Vol. 72 ›› Issue (6): 720-730.DOI: 10.6023/A14040274 上一篇    下一篇

研究论文

新型双β-环糊精键合SBA-15液相手性固定相的制备及评价

周仁丹, 李来生, 程彪平, 聂桂珍, 张宏福   

  1. 南昌大学分析测试中心 南昌 330047
  • 收稿日期:2014-04-14 出版日期:2014-06-14 发布日期:2014-05-11
  • 通讯作者: 李来生 E-mail:lilaishengcn@163.com
  • 基金资助:
    项目受国家自然科学基金(No.21165012)、江西省自然科学基金(No.2010GZH0089)、江西省教育厅科技项目(No.GJJ11274)资助.

Preparation and Evaluation of a Novel bis(β-cyclodextrin)-bonded SBA-15 Chiral Stationary Phase for HPLC

Zhou Rendan, Li Laisheng, Cheng Biaoping, Nie Guizhen, Zhang Hongfu   

  1. Center of Analysis and Testing, Nanchang University, Nanchang 330047
  • Received:2014-04-14 Online:2014-06-14 Published:2014-05-11
  • Supported by:
    Project supported by the Natural Science Foundation of China (No.21165012), the Natural Science Foundation of Jiangxi Province (No.2010GZH0089) and Jiangxi Province Education Commission Foundation (No.GJJ11274).

由于桥联双β-环糊精的协同包结作用和独特的多重识别功能已被广泛地用作人工模拟酶,本文尝试用桥联双β-环糊精作固定相配体,开发其手性分离功能.采用3-异氰酸丙基三乙氧基硅烷偶联剂连续反应法,首先将(6-氧-对甲苯磺酰)-β-环糊精键合到有序介孔SBA-15硅胶表面,然后与(6-乙二胺-6-去氧)-β-环糊精反应,制备一种新型的乙二胺桥联双β-环糊精键合SBA-15液相色谱手性固定相(BCDSP).采用质谱、红外光谱、元素分析、热重分析和透射电镜等进行结构表征.在极性有机溶剂模式下,评价了新固定相的基本色谱性能,并用于β-受体阻滞剂对映体的拆分.考察了流动相中有机溶剂、三乙胺、冰醋酸改性剂用量和柱温对手性分离的影响.基于优化的色谱条件,采用双β-环糊精键合固定相成功地拆分了常用的14种β-受体阻滞剂药物对映体,其中普萘洛尔的分离度可达到2.18,卡维地洛的分离度可达到2.01,分析时间一般为10~20 min,取得了较高的分离效率.为比较研究,还制备了一种单β-环糊精固定相(CDSP),结果发现双β-环糊精键合相可拆分14种β-受体阻滞剂,而在优化的条件下单β-环糊精键合相仅能部分拆分4种β-受体阻滞剂,且前者的分离度明显优于后者.基于实验数据对双β-环糊精和单β-环糊精固定相的分离机理的异同点展开讨论.一方面包结作用和氢键作用在桥联双β-环糊精键合相的手性分离中占重要地位,与常见的单β-环糊精固定相相似;另一方面桥联双β-环糊精固定相拥有自身的特点,两个β-环糊精腔体存在协同作用,相应于它的模拟酶功能,协同作用不仅扩大了空间识别域,还提高了立体选择性,基于双腔体的包结作用和乙二胺桥键的氢键作用等增强了固定相的手性识别能力,加上有序的SBA-15能加快传质,使得BCDSP具有较好的手性色谱性能,拥有更广泛的拆分对象、更高的分离度和更短的分析时间.新固定相的制备方法简便,手性分离功能强,色谱性能稳定,且制备成本较低.这类桥联双β-环糊精键合有序介孔SBA-15新型的快速分离材料在手性药物质量监控和药代动力学研究中存在良好的应用前景.

关键词: 高效液相色谱法, 乙二胺桥联双β-环糊精键合SBA-15手性固定相, 协同分离机理, β-受体阻滞剂, 手性药物

The bridged β-cyclodextrin has been widely used as artificial enzymes due to its synergistic inclusion effect and unique multi-recognition function. In this paper, we employed bridged β-cyclodextrin as ligand in order to explore its chiral separation function. A novel N,N'-ethylenediamino bridged bis(β-cyclodextrin)-bonded SBA-15 chiral stationary phase (BCDSP) for HPLC was first developed by solid successive reaction method. In the first step, a 6-O-toluene-sulfonyl-β- cyclodextrin was bonded to ordered mesoporous SBA-15 by using 3-isocyanatopropyltriethoxysilane as coupling reagent. Then the bonded SBA-15 silica continued to react with mono-(6-ethylenediamino-6-deoxy)-β-cyclodextrin and obtained the BCDSP with bridged double β-cyclodextrin ligand. Its chemical structure was characterized by mass spectrometry, infrared spectroscopy, elemental analysis, thermogravimetric analysis and transmission electron microscopy. The basic chromatographic property of new stationary phase was evaluated in polar organic mode and used for the separations of β-blockers enantiomers. Some effect factors such as the concentrations of organic solvent, triethylamine (TEA) and glacial acetic acid (HOAc) in mobile phase, column temperature on the resolution were investigated. The 14 kinds of β-blockers enantiomers on BCDSP column were successfully separated under the optimized chromatographic conditions, in which the resolutions of propranolol enantiomers and carvedilol enantiomers were 2.18 and 2.01 within 10~20 min, respectively. By comparative study, the bis(β-cyclodextrin)-bonded phase could separate all 14 kinds of β-blockers enantiomers, while native β-cyclodextrin-bonded phase (CDSP) with single ring ligand could partially resolve about 4 kinds of them under optimized condition. Moreover, the resolutions of drugs on BCDSP were more than those on native β-cyclodextrin column. The chromatographic data analysis showed that BCDSP with double β-cyclodextrin ligand exhibited a stronger enantioseparation ability and wider analytes as comparison with the native β-cyclodextrin stationary phase. The excellent property of BCDSP should be due to the double β-cyclodextrin ligand expands the chiral recognization for enantiomers of drugs. Meanwhile, the inclusion interaction from double cavities and the hydrogen bonding interaction from the bridged ethylenediamino group with the tested enantiomers improved the chiral selectivities of β-blockers. The ordered pore of SBA-15 could also reduce the mass transfer resistance and enhance the chiral chromatographic property of BCDSP. According to chromatographic data, the similarities and differences of separation mechanism of double β-cyclodextrin and single β-cyclodextrin stationary phases were discussed. On the one hand, the inclusion complexing and hydrogen bonding interactions of BCDSP played important roles in chiral separations, which was similar to the common single-β-cyclodextrin stationary phase. On the other hand, the synergistic interaction of double β-cyclodextrin cavities could expand spatial recognition domain, making BCDSP with better chiral chromatographic performance, wider analytes, higher resolution and shorter analysis time. This new stationary phase has clear and stable structure of ligand, convenient preparation method, lower cost, especially, its excellent enantioseparation ability. The exploration of this double β-cyclodextrin-bonded phase could provide a kind of new type separation material for wide chiral drugs. It has a research significance and bright prospect in quality-controlling of chiral drugs.

Key words: high performance liquid chromatography, ethylenediamino bridged bis(β-cyclodextrin)-bonded SBA-15 chiral stationary phase, synergistic separation mechanism, β-blockers, chiral drugs