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金纳米粒子修饰手性毛细管电色谱固定相的制备及性能表征

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

作者信息

Xiong Lele, Li Ruijun, Ji Yibing

机构信息

College of Science, China Pharmaceutical University, Nanjing 210009, China.

出版信息

Se Pu. 2017 Jul 8;35(7):712-718. doi: 10.3724/SP.J.1123.2017.03019.

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.

摘要

制备了金纳米颗粒(GNPs,15纳米)并将其引入到氨基衍生的硅胶整体柱中。通过共价修饰方法将牛血清白蛋白(BSA)固定在羧基功能化的GNPs上,以提供用于对映体分离的手性固定相(CSP)。通过透射电子显微镜(TEM)、紫外(UV)-可见吸收光谱和扫描电子显微镜(SEM)等表征方法,GNPs得到了良好的分散,并成功地负载到柱上,负载量高达17.18%。优化了BSA修饰CSP的制备条件,选择10%(v/v)的3-氨丙基三乙氧基硅烷(APTES)和15 g/L的BSA作为合适的反应条件。通过毛细管电色谱(CEC)研究了BSA修饰CSP的对映体分离性能。色氨酸、麻黄碱和阿替洛尔的对映体得到了分离,实现了色氨酸的基线分离。同时,研究了pH值、缓冲液浓度和施加电压对手性分离的影响,最佳分离条件为pH 7.4的10 mmol/L磷酸盐缓冲液和15 kV的施加电压。与通过物理吸附制备的BSA修饰CSP相比,通过共价修饰方法制备的CSP具有更好的分离效果,分析物无需柱前衍生即可直接分离。此外,制备的BSA修饰CSP表现出良好的批间重复性,迁移时间和选择性因子的相对标准偏差(RSDs)分别不超过2.3%和0.96%。这项工作为用其他蛋白质或其他类型手性选择剂进行修饰提供了很好的思路。

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