Schlemmer Bettina, Bandari Rajendar, Rosenkranz Lutz, Buchmeiser Michael R
Leibniz-Institut für Oberflächenmodifizierung e.V., Permoserstrasse 15, D-04318 Leipzig, Germany.
J Chromatogr A. 2009 Mar 27;1216(13):2664-70. doi: 10.1016/j.chroma.2008.09.003. Epub 2008 Sep 5.
Monolithic capillary columns were prepared via electron beam triggered free radical polymerization within the confines of 0.2 and 0.1mm I.D. capillary columns using ethyl methacrylate and trimethylolpropane triacrylate as monomers as well as 2-propanol, 1-dodecanol and toluene as porogenic system. The influence of column diameter on reproducibility and separation performance was investigated. For evaluation, a protein standard consisting of five proteins in the range of 5800-66,000 g mol(-1) was used. Reproducibility was checked by determining the relative standard deviations in retention times, peak widths at half height, asymmetry and resolution. Excellent run-to-run reproducibility was found for both 0.2 and 0.1mm I.D. columns; batch-to-batch reproducibility was good for both column types. In order to enhance the non-polar character of the monolithic columns, lauryl methacrylate-based capillary columns were prepared. These were successfully used for the separation of proteins and a cytochrome c digest.
通过电子束引发自由基聚合反应,在内径为0.2毫米和0.1毫米的毛细管柱内制备整体式毛细管柱,使用甲基丙烯酸乙酯和三羟甲基丙烷三丙烯酸酯作为单体,以及2-丙醇、1-十二醇和甲苯作为致孔体系。研究了柱直径对重现性和分离性能的影响。为了进行评估,使用了一种由五种分子量在5800 - 66,000 g mol(-1)范围内的蛋白质组成的蛋白质标准品。通过测定保留时间、半高峰宽、不对称度和分离度的相对标准偏差来检查重现性。对于内径为0.2毫米和0.1毫米的柱子,均发现了出色的批内重现性;两种柱型的批间重现性都很好。为了增强整体柱的非极性特征,制备了基于甲基丙烯酸月桂酯的毛细管柱。这些柱子成功用于蛋白质和细胞色素c消化产物的分离。