Protein separation using a novel silica-based RPLC/IEC stationary phase modified with N-methylimidazolium ionic liquid |
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Authors: | Yi-Xin Wang Kai-Lou Zhao Fan Yang Lei Tian Ying Yang Quan Bai |
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Institution: | Key Laboratory of Synthetic and Natural Functional Molecule Chemistry of Ministry of Education, Key Laboratory of Modern Separation Science of Shaanxi Province, Institute of Modern Separation Science, Northwest University, Xi'an 710069, China |
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Abstract: | 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 (SilprMim) 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. |
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Keywords: | Ionic liquid Mixed-mode chromatography Protein separation Reversed-phase liquid chromatography Ion-exchange chromatography |
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