Growth of two-layer copolymer as the stationary phase with very high separation efficiency for separating peptides in capillary electrochromatography |
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Authors: | Genlin Sun Weiyang Tang Yao Lu Kyung Ho Row |
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Affiliation: | 1. Department of Pharmacy, Jiangxi University of Chinese Medicine, Nanchang, P. R. China;2. Department of Chemistry, Inha University, Incheon, South Korea |
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Abstract: | 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. |
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Keywords: | Capillary electrochromatography Open tubular capillary column Peptides RAFT polymerization van Deemter |
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