色谱 ›› 2017, Vol. 35 ›› Issue (7): 712-718.DOI: 10.3724/SP.J.1123.2017.03019

• 研究论文 • 上一篇    下一篇

金纳米粒子修饰的手性毛细管电色谱固定相的制备及性能表征

熊乐乐, 李瑞军, 季一兵   

  1. 中国药科大学理学院, 江苏 南京 210009
  • 收稿日期:2017-03-07 出版日期:2017-07-08 发布日期:2013-10-18
  • 通讯作者: 季一兵,Tel:(025)86185150,E-mail:jiyibing@msn.com

Preparation and performance characterization of gold nanoparticles modified chiral capillary electrochromatography stationary phase

XIONG Lele, LI Ruijun, JI Yibing   

  1. College of Science, China Pharmaceutical University, Nanjing 210009, China
  • Received:2017-03-07 Online:2017-07-08 Published:2013-10-18

摘要:

制备了粒径为15 nm的金纳米粒子(GNPs)并将其修饰到氨基衍生化的硅胶整体柱内,通过化学键合法将牛血清白蛋白(BSA)固载到GNPs的表面作为手性固定相。通过透射电子显微镜、扫描电子显微镜等方法进行表征,结果表明,GNPs分散性良好,并被成功地修饰到毛细管柱内,含量高达17.18%。优化了BSA手性柱的制备条件,最终确定了体积分数为10%的3-氨丙基三乙氧基硅烷(APTES)和15 g/L BSA为最佳反应条件。在毛细管电色谱分离模式下,对缓冲液pH值、电压等分离条件进行了考察,最终选择了10 mmol/L pH 7.4的磷酸缓冲液和15 kV运行电压作为最佳分离条件。手性柱对3种手性化合物(色氨酸、阿替洛尔和麻黄碱)有拆分效果,对色氨酸能实现基线分离。与物理吸附法相比,化学键合法制备的手性柱拆分效果好,分析物无需柱前衍生化,且色谱柱稳定性良好。该文的制备方法也为其他类型手性选择剂的引入提供了良好的思路。

关键词: 硅胶整体柱, 金纳米粒子, 毛细管电色谱, 手性固定相

Abstract:

Gold nanoparticles (GNPs, 15 nm) were prepared and introduced to amino groups derived silica monolithic column. Bovine serum albumin (BSA) was immobilized via covalent modification method onto the carboxylic functionalized GNPs to afford chiral stationary phase (CSP) for enantioseparation. GNPs were well dispersed and successfully incorporated onto the columns with the contents as high as 17.18% by characterization method such as transmission electron microscopy (TEM), ultraviolet (UV)-visible absorption spectra and scanning electron microscopy (SEM). The preparation conditions of the BSA modified CSP were optimized and 10% (v/v) 3-aminopropyltriethoxysilane (APTES) and 15 g/L BSA were selected as appropriate reaction conditions. The enantioseparation performance of the BSA modified CSP has been investigated by capillary electrochromatography (CEC). Enantiomers of tryptophan, ephedrine and atenolol were resolved, and the baseline separation of tryptophan was achieved. Meanwhile, the influences of pH value, buffer concentrations and applied voltages used on the chiral separation were studied, and the optimal separation conditions were 10 mmol/L phosphate buffer at pH 7.4 and 15 kV applied voltages. In comparison with the BSA modified CSP prepared by physical adsorption, the CSP prepared by covalent modification method had better separation results, and the analytes could be separated directly without pre-column derivatization. In addition, the prepared BSA modified CSP exhibited good run to run repeatability with relative standard deviations (RSDs) of the migration times and selectivity factors not more than 2.3% and 0.96%, respectively. This work offers a good thinking for modification with other proteins or other types of chiral selectors.

Key words: capillary electrochromatography (CEC), chiral stationary phase (CSP), gold nanoparticles (GNPs), silica monolithic column (CSM)

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