首页> 外文期刊>Electrophoresis: The Official Journal of the International Electrophoresis Society >Capillary electrophoresis-mass spectrometry of basic proteins using a new physically adsorbed polymer coating. Some applications in food analysis
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Capillary electrophoresis-mass spectrometry of basic proteins using a new physically adsorbed polymer coating. Some applications in food analysis

机译:使用新的物理吸附的聚合物涂层进行碱性蛋白质的毛细管电泳-质谱分析。在食品分析中的一些应用

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摘要

A new physically adsorbed capillary coating for capillary electrophoresis-mass spectrometry (CE-MS) of basic proteins is presented, which is easily obtained by flushing the capillary with a polymer aqueous solution for two min. This coating significantly reduces the electrostatic adsorption of a group of basic proteins (i.e., cytochrome c, lysozyme, and ribonuclease A) onto the capillary wall allowing their analysis by CE-MS. The coating protocol is compatible with electrospray inonization (ESI)-MS via the reproducible separation of the standard basic proteins (%RSD values (n = 5) < 1% for analysis time reproducibility and < 5% for peak heights, measured from the total ion electropherograms (TIEs) within the same day). The LODs determined using cytochrome c with total ion current and extracted ion current defection were 24.5 and 2.9 fmol, respectively. Using this new coating lysozymes from chicken and turkey egg white could be easily distinguished by CE-MS, demonstrating the usefulness of this method to differentiate animal species. Even after sterilization at 120 degreesC for 30 min, lysozyme could be detected, as well as in wines at concentrations much lower than the limit marked by the EC Commission Regulation. Adulteration of minced meat with 5% of egg-white could also be analysed by our CE-MS protocol.
机译:提出了一种用于碱性蛋白毛细管电泳-质谱(CE-MS)的新型物理吸附毛细管涂层,通过用聚合物水溶液冲洗毛细管两分钟即可轻松获得。这种涂层显着减少了一组碱性蛋白(即细胞色素c,溶菌酶和核糖核酸酶A)在毛细管壁上的静电吸附,从而可以通过CE-MS进行分析。通过可重现的标准碱性蛋白质分离,涂层协议与电喷雾离子化(ESI)-MS兼容(%RSD值(n = 5),分析时间重现性<1%,峰高<5%,从总测量值离子电泳图(TIE))。使用细胞色素c测得的总离子流和萃取离子流缺陷的LOD分别为24.5和2.9 fmol。 CE-MS可以很容易地使用鸡和火鸡蛋白的这种新型包膜溶菌酶,从而证明了该方法对区分动物物种的有用性。即使在120摄氏度下灭菌30分钟后,也可以检测到溶菌酶,并且葡萄酒中的溶菌酶浓度也远远低于EC委员会法规规定的限值。碎肉与5%的蛋清的掺假也可以通过我们的CE-MS协议进行分析。

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