Surface-alkylated polystyrene monolithic columns for peptide analysis in capillary liquid chromatography-electrospray ionization mass spectrometry.
Surface-alkylated polystyrene monolithic columns for peptide analysis in capillary liquid chromatography-electrospray ionization mass spectrometry.
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DOI：
10.1021/ac011202w
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发表时间：
2002-04
影响因子：
7.4
通讯作者：
Xian Huang;S. Zhang;G. Schultz;J. Henion
中科院分区：
文献类型：
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作者：
Xian Huang;S. Zhang;G. Schultz;J. Henion
大孔聚（苯乙烯-二乙烯基苯）（PS-DVB）整料通过在PEEK、熔融石英或内径为75或125微米的不锈钢管中原位聚合来制备。描述了用于随后烷基化整料的流动接触表面的方法。该方法用含有溶解的Friedel-Crafts催化剂、烷基卤（1-氯十八烷）和有机溶剂的溶液处理包括刚性大孔整料的通孔表面在内的所有表面。与未修饰的整体PS-DVB柱相比，该方法产生了改进的肽的反相液相色谱分离。表面十八烷基化对于蛋白质的反相分离是不必要的，因为未改性和改性的柱都提供相当的结果。采用电喷雾质谱检测，胰蛋白酶蛋白酶，标准蛋白质和标准肽用于评价整体柱。利用烷基化整体柱结合纳米制造的电喷雾离子化芯片进行蛋白质组学研究的潜在应用也进行了讨论。
Macroporous poly(styrene-divinylbenzene) (PS-DVB) monoliths were prepared by in situ polymerization in PEEK, fused silica, or stainless steel tubing having an inner diameter of 75 or 125 microm. A process is described for subsequent alkylation of the flow-contacting surfaces of the monoliths. The process treats all the surfaces including through-pore surfaces of the rigid macroporous monolith with a solution containing a dissolved Friedel-Crafts catalyst, an alkyl halide (1-chlorooctadecane), and an organic solvent. This process produces an improved reversed-phase liquid chromatographic separation of peptides compared to an unmodified monolithic PS-DVB column. The surface octadecylation is not necessary for a reversed-phase separation of proteins since both unmodified and modified columns provide comparable results. Tryptic protein digests, standard proteins, and standard peptides were used to evaluate the monolithic columns by employing electrospray mass spectrometry detection. Potential applications in proteomics studies by mass spectrometry, which use the alkylated monolithic column engaged onto the nanofabricated electrospray ionization chip, are also discussed.