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亲和毛细管整体柱在牛血清白蛋白与奈福泮对映体相互作用研究中的应用

曾玙, 季一兵

曾玙, 季一兵. 亲和毛细管整体柱在牛血清白蛋白与奈福泮对映体相互作用研究中的应用[J]. 中国药科大学学报, 2016, 47(1): 66-72. DOI: 10.11665/j.issn.1000-5048.20160109
引用本文: 曾玙, 季一兵. 亲和毛细管整体柱在牛血清白蛋白与奈福泮对映体相互作用研究中的应用[J]. 中国药科大学学报, 2016, 47(1): 66-72. DOI: 10.11665/j.issn.1000-5048.20160109
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ZENG Yu, JI Yibing. Study of interaction between bovine serum albumin and nefopam enantiomers with affinity capillary monolith[J]. Journal of China Pharmaceutical University, 2016, 47(1): 66-72. DOI: 10.11665/j.issn.1000-5048.20160109
Citation: ZENG Yu, JI Yibing. Study of interaction between bovine serum albumin and nefopam enantiomers with affinity capillary monolith[J]. Journal of China Pharmaceutical University, 2016, 47(1): 66-72. DOI: 10.11665/j.issn.1000-5048.20160109
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亲和毛细管整体柱在牛血清白蛋白与奈福泮对映体相互作用研究中的应用

  • 中国药科大学分析化学教研室

Study of interaction between bovine serum albumin and nefopam enantiomers with affinity capillary monolith

摘要

    摘要
  • 摘要: 利用胃蛋白酶键合的有机聚合物整体柱在电色谱中对手性药物奈福泮的拆分能力，采用前沿分析法同时对奈福泮的两个对映体与牛血清白蛋白(BSA)的相互作用情况进行了考察。经过优化后建立的电色谱条件为:胃蛋白酶修饰的聚(甲基丙烯酸环氧丙酯-乙二醇二甲基丙烯酸酯)毛细管整体柱作为分离通道(32 cm×75 μm，有效长度22 cm)，运行缓冲液为pH 5.5的15 mmol/L醋酸铵，样品溶剂为pH 7.4的50 mmol/L醋酸铵，运行电压为5.0 kV，电压进样10 kV×6 s，检测波长为215 nm。此时奈福泮两个对映体平台峰彼此完全分离，结合体系中BSA对对映体在电色谱中的分离和检测均无影响，测得两个对映体与BSA的结合常数分别为443和527 L/mol，结合位点数均为1.0，结合位点为Sudlow site Ⅱ。
    Abstract: A pepsin modified poly(glycidyl methacrylate-ethyleneglycol dimethacrylate)(poly(GMA-EDMA))capillary monolith(32 cm×75 μm, effective lenth 22 cm)was applied in exploring the interaction between nefopam enantiomers and bovine serum albumin(BSA). Frontal analysis was used to measure the binding constant, number of binding sites and the location of binding sites of BSA to both nefopam enantiomers. Optimal CEC conditions were as follows: 15 mmol/L ammonium acetate was adjusted to pH 5. 5 as running buffer, separation voltage was 5. 0 kV, detection wavelength was 215 nm, and samples were dissolved with 50 mmol/L ammonium acetate buffer at pH 7. 4 and were injected at 10 kV×6 s. The results indicated that two enantiomers could successfully separated by CEC on the monolith column. BSA in the binding system showed no effect on the separation and determination of nefopam enantiomers. Frontal analysis demonstrated that BSA has only one binding site with both enantiomers, with binding constants(K)of 443 L/mol and 527 L/mol, respectively. The displacement expe-riments indicated that binding site of both isomers to BSA molecule was Sudlow siteⅡ.
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  • [1] Zheng Y,Wang X,Ji YB.Monoliths with proteins as chiral selectors for enantiomer separation[J].Talanta,2012,91:7-17.
    [2] Chi CJ, Wang W, Ji YB. Preparation and evaluation of pepsin affinity organic polymer capillary monolithic column[J].Chin J Chromatogr(色谱),2014,32(8):791-797.
    [3] Hong TT,Chi CJ,Ji YB.Pepsin-modified chiral monolithic column for affinity capillary electrochromatography[J].J Sep Sci,2014,37(22):3377-3383.
    [4] Hong TT,Zheng Y,Hu WW,et al.Preparation and evaluation of bovine serum albumin immobilized chiral monolithic column for affinity capillary electrochromatography[J].Anal Biochem,2014,464:43-50.
    [5] He J,Ji YB.Proteins as chiral selectors for enantiomer separation[J].Pharm Clin Res(药学与临床研究),2013,21(2):166-171.
    [6] Girardot M,Li HY,Descroix S,et al.Determination of binding parameters between lysozyme and its aptamer by frontal analysis continuous microchip electrophoresis(FACMCE)[J].J Chromatogr A,2011,1218(26):4052-4058.
    [7] Sisavath N, Leclercq L, Le Saux T, et al. Study of interactions between oppositely charged dendrigraft poly-L-lysine and human serum albumin by continuous frontal analysis capillary electrophoresis and fluorescence spectroscopy[J].J Chromatogr A,2013,1289:127-132.
    [8] Zhao LJ, Chen DY. Characterization of interactions between methoxatin disodium salt and human serum albumin by pressure-assisted capillary electrophoresis/frontal analysis and circular dichroism spectroscopy[J].Biomed Chromatogr,2015,29(1):123-128.
    [9] Liu C,Kang JW.Improved capillary electrophoresis frontal analysis by dynamically coating the capillary with polyelectrolyte multilayers[J].J Chromatogr A,2012,1238:146-151.
    [10] Busch MH,Carels LB,Boelens HF,et al.Comparison of five methods for the study of drug-protein binding in affinity capillary electrophoresis[J].J Chromatogr A,1997,777(2):311-328.
    [11] Østergaard J,Schou C,Larsen C,et al.Evalution of capillary electrophoresis-frontal analysis for the study of low molecular weight drug-human serum albumin interactions[J].Electrophoresis,2002,23(17):2842-2853.
    [12] Tu FZ,Xiang XY,Dong Y,et al.Studies on the interaction for bovine serum albumin with benzoate by capillary zone electrophoresis frontal analysis[J].Chin J Anal Lab(分析试验室),2011,30(8):73-76.
    [13] Jiang P,Wu LQ,Rena B,et al.Study of interaction of fluorescein sodium and bovine serum albumin by capillary electrophoresis/frontal analysis and molecular modeling technology[J].Chin J Anal Chem(分析化学),2011,39(5):680-684.
    [14] Mohamed NA,Kuroda Y,Shibukawa A,et al.Enantioselective binding analysis of verapamil to plasma lipoproteins by capillary electrophoresis-frontal analysis[J].J Chromatogr A,2000,875(1/2):447-453.
    [15] Ohara T,Shibukawa A,Nakagawa T.Capillary electrophoresis/frontal analysis for microanalysis of enantioselective protein binding of a basic drug[J].Anal Chem,1995,67(19):3520-3525.
    [16] Ding YS, Zhu XF, Lin BC. Study of interaction between drug enantiomers and serum albumin by capillary electrophoresis[J].Electrophoresis,1999,20(9):1890-1894.
    [17] Ishihama Y,Miwa T,Asakawa N.Drug-plasma protein binding assay by electrokinetic chromatography-frontal analysis[J].Electrophoresis,2002,23(6):951-955.
    [18] Shiono H,Shibukawa A,Kuroda Y,et al.Effect of sialic acid residues of human alpha 1-acid glycoprotein on stereoselectivity in basic drug-protein binding[J].Chirality,1997,9(3):291-296.
    [19] Ding YS,Zhu XF,Lin BC.Albumin-drug binding study by capillary electrophoresis Ⅱ.Determination of drug enantiomeric binding constants[J].Chin J Chromatogr(色谱),1999,17(2):138-141.
    [20] Ding YS,Lin BC.Investigation of molecular association by capillary zone electrophoresis / frontal analysis[J].Chin J Anal Chem(分析化学),1999,27(1):20-23.
    [21] Martínez-Pla JJ,Martínez-Gómez MA,Martín-Biosca Y,et al.High-throughput capillary electrophoresis frontal analysis method for the study of drug interactions with human serum albumin at near-physiological conditions[J].Electrophoresis,2004,25(18/19):3176-3185.
    [22] Liu Y,Liu YM,Xia ZN.Investigation of interaction of potassium ion to 18-crown-6 by capillary electrophoresis / frontal analysis[J].Chin J Anal Chem(分析化学),2003,31(12):1500-1503.
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  • 刊出日期:  2016-02-24

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