Open tubular CEC (OT-CEC) column with a very high separation efficiency was prepared for peptides separation. A pretreated silica-fused capillary was reacted with 3-(methacryloxy) propyltrimethoxysilane followed by vinylbenzyl chloride and divinylbenzene to produce first thin monolithic monolayer. The second copolymer layer was formed on thin monolithic monolayer of the capillary by reversible addition-fragmentation transfer polymerization of N-phenylacrylamide and styrene. The key parameters including buffer pH value and organic modifier were systematically evaluated to provide the optimal chromatographic condition. The resultant OT-CEC columns were validated by separating a synthetic mixture of peptides and cytochrome C tryptic digest in capillary electrochromatography. The number of theoretical plates as high as 2.4 million per column was achieved for synthetic mixture peptides. In addition, the fabricated OT-CEC column also resolved more than 18 high-efficiency digestion peptides from a mixture containing tryptic digest of cytochrome C. The column to column and inter- to intraday repeatabilities of OT-CEC column through RSD% were found better than 3.0%, exhibiting satisfactory stability and repeatability of the two-layer deposited OT-CEC column. The results reveal that the open tubular capillary column modified with two-layer copolymer shows the great prospect for the separation of proteins in capillary electrochromatography.

中文翻译：

---

两层共聚物作为固定相的生长在毛细管电色谱中分离肽的分离效率非常高

制备了具有非常高分离效率的开管式 CEC (OT-CEC) 柱用于肽分离。预处理的石英熔融毛细管与 3-(甲基丙烯酰氧基) 丙基三甲氧基硅烷反应，然后与乙烯基苄基氯和二乙烯基苯反应，以产生第一个薄的整体单层。通过N的可逆加成-断裂转移聚合在毛细管的单片薄单层上形成第二共聚物层-苯基丙烯酰胺和苯乙烯。系统评价了缓冲液pH值和有机改性剂等关键参数，以提供最佳的色谱条件。通过在毛细管电色谱中分离肽和细胞色素 C 胰蛋白酶消化物的合成混合物来验证所得的 OT-CEC 柱。合成混合肽的理论塔板数高达每柱 240 万个。此外，制造的 OT-CEC 色谱柱还从含有细胞色素 C 胰蛋白酶消化物的混合物中分离出超过 18 种高效消化肽。通过 RSD% 发现 OT-CEC 色谱柱的柱间和日间重复性更好小于 3.0%，表明两层沉积 OT-CEC 色谱柱的稳定性和重复性令人满意。