Capillary electrochromatography of proteins and peptides with a cationic acrylic monolith.
Capillary electrochromatography of proteins and peptides with a cationic acrylic monolith.
复制标题
使用阳离子丙烯酸整料对蛋白质和肽进行毛细管电色谱分析。
DOI：
10.1016/s0021-9673(00)00250-8
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发表时间：
2000
期刊：
影响因子：
--
通讯作者：
Horváth,C
中科院分区：
文献类型：
--
作者：
Zhang,S;Huang,X;Zhang,J;Horváth,C
用50 μm内径的硅烷化熔融石英毛细管制备了毛细管电色谱整体固定相，用于蛋白质和多肽的分离。通过甲基丙烯酸缩水甘油酯、甲基丙烯酸甲酯和乙二醇二甲基丙烯酸酯在丙醇和甲酰胺作为致孔剂存在下的原位共聚。在多孔整料的表面处的环氧基团与N-乙基丁胺反应以形成具有乙基链和丁基链的固定叔氨基官能团。核糖核酸酶A、胰岛素、α-乳白蛋白和肌红蛋白的混合物通过反方向CEC等度分离，水有机移动的流动相含有乙腈和磷酸钠缓冲液，pH 2.5。在相似条件下，也实现了四种血管紧张素型肽的CEC分离。蛋白质的洗脱顺序与反相色谱中获得的顺序相似。蛋白质和肽的迁移因子与乙腈浓度的关系图显示出相反的趋势。这很可能是由于在反向CEC系统中，蛋白质的色谱保留比肽大，电泳迁移速度比肽低。由此可以得出结论，分离是由一个双重机制，涉及选择性色谱保留和差分电泳迁移之间的复杂的相互作用。
For the separation of proteins and peptides by capillary electrochromatography (CEC), columns with a monolithic stationary phase were prepared from silanized fused-silica capillaries of 50 μm I.D. by in situ copolymerization of glycidyl methacrylate, methyl methacrylate and ethylene glycol dimethacrylate in the presence of propanol and formamide as porogens. The epoxide groups at the surface of the porous monolith were reacted with N-ethylbutylamine to form fixed tertiary amino functions with ethyl- and butyl-chains. A mixture of ribonuclease A, insulin, α-lactalbumin and myoglobin was separated isocratically by counterdirectional CEC with hydro–organic mobile phases containing acetonitrile and sodium phosphate buffer, pH 2.5. The separation of four angiotensin type peptides by CEC was also achieved under similar conditions. The elution order of proteins was similar to that obtained in reversed-phase chromatography. Plots of the migration factors for proteins and peptides against the acetonitrile concentration exhibit opposite trends. This is most likely due to the greater chromatographic retention and lower electrophoretic migration velocity of proteins than that of peptides in the counterdirectional CEC system. From this it is concluded that the separation is governed by a dual mechanism that involves the complex interplay between selective chromatographic retention and differential electrophoretic migration.