新型共聚物涂层毛细管电泳柱及其分离蛋白质的研究

The Novel Copolymer Coated Capillary Columns of Electrophoresis and Their Applications to Separation of Proteins

研究新型共聚物——ＺＢ系列表面键合剂在毛细管电泳中的应用。采用物理吸附的方法制备了ＺＢ－００４，ＺＢ－０１４，ＺＢ－０１６等３种涂层毛细管柱，在ｐＨ３～５范围内，３种涂层均能有效地降低管壁对蛋白质的吸附作用和电渗流，其中亲水性较弱的ＺＢ－００４涂层的分离性能最好。在ｐＨ＜５时，涂层具有较高的稳定性和良好的分析重复性，但在更高的ｐＨ值条件下，仍然存在着峰形畸变和电渗流迅速增加的现象。

The copolymer of acrylonitrile, methyl acrylate, hydroxy ethyl acrylate (ZB 004), the copolymer of acrylonitrile, methyl acrylate, hydroxy ethyl acrylate, acrylamide (ZB 014) and the copolymer of acrylonitrile, hydroxy ethyl acrylate (ZB 016) were coated on the inner surface of fused silica capillaries by just filling the capillary with solutions containing these copolymers followed by flushing the capillary with nitrogen. The physically adsorbed layer can reduce both protein adsorption and electroosmotic flow in the pH range of 3 5. Electroosmotic flow descreased by raising the concentrations of the copolymers. Separation performance of ZB 004 layer is better than those of other two layers due to its low hydrophilicity, but with higher pH values, appreciable peak deformation and increase in electroosmosis were observed. The intra day and inter day migration reproducibility were investigated in terms of relative standard deviation ( RSD ) with four basic proteins at pH 4 0. The RSD s of the intra day migration times were less than 2%. The RSD s of the inter day migration times were less than 4%. At pH 5 0, the RSD s of the migration times in two ZB 004 coated capillaries made on two different days were less than 1%. Separation efficiencies of four basic proteins in a ZB 004 coated capillary which stored in a buffer (pH 4 0) for fifteen days after being used for 14 days decreased 15%. These coatings were stable and exhibited reproducible separations from intra day, inter day and inter column under acidic conditions.