Protein separation using a novel silica-based RPLC/IEC stationary phase modified with N-methylimidazolium ionic liquid

• Published in: Chinese chemical letters Vol. 26; no. 8; pp. 988 - 992
• Main Authors: Wang, Yi-Xin, Zhao, Kai-Lou, Yang, Fan, Tian, Lei, Yang, Ying, Bai, Quan
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 01.08.2015
• Subjects: IEC; Ion-exchange chromatography; Ionic liquid; Mixed-mode chromatography; N-甲基咪唑; Protein separation; Reversed-phase liquid chromatography; 反相高效液相色谱; 固定相; 硅胶; 离子液体; 蛋白质分离; 高效液相色谱法; Protein separation; Reversed-phase liquid chromatography; Mixed-mode chromatography; Ion-exchange chromatography; Ionic liquid
• ISSN: 1001-8417; 1878-5964
• DOI: 10.1016/j.cclet.2015.05.001

Ionic liquids（ILs） immobilized on silica as novel high performance liquid chromatography（HPLC）stationary phases have attracted considerable attention. However, it has not been applied to protein separation. In this paper, N-methylimidazolium IL-modified silica-based stationary phase（Silpr Mim）was prepared and investigated as a novel multi-interaction stationary phase charged positively for protein separation. The results indicate that all of the basic proteins tested cannot be absorbed on this novel stationary phase, whereas all of the acidic proteins tested can be retained, and the baseline separation of eight kinds of acidic protein standards can be achieved when performed in reversed phase/ion-exchange chromatography（RPLC/IEC） mode. Compared with commonly used commercial octadecylated silica（ODS） column, the novel stationary phase can show selectivity and good resolution to acidic proteins, which has a promising application in the separation and analyses of acidic proteins from the complex samples in proteomics. In addition, the chromatographic behavior of proteins, the effect of the ligand structure and the retention mechanism on this stationary phase were also investigated.