Premium
Separation of peptides on mixed mode of reversed‐phase and ion‐exchange capillary electrochromatography with a monolithic column
Author(s) -
Wu Ren’an,
Zou* Hanfa,
Fu Hongjing,
Jin Wenhai,
Ye Mingliang
Publication year - 2002
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(200205)23:9<1239::aid-elps1239>3.0.co;2-x
Subject(s) - capillary electrochromatography , electrochromatography , chromatography , monolithic hplc column , phase (matter) , ion exchange , capillary action , chemistry , column (typography) , materials science , ion , analytical chemistry (journal) , capillary electrophoresis , stationary phase , computer science , organic chemistry , composite material , telecommunications , frame (networking)
The mixed mode of reversed phase (RP) and strong cation‐exchange (SCX) capillary electrochromatography (CEC) based on a monolithic capillary column has been developed. The capillary monolithic column was prepared by in situ copolymerization of 2‐(sulfooxy)ethyl methacrylate (SEMA) and ethylene dimethacrylate (EDMA) in the presence of porogens. The sulfate group provided by the monomer SEMA on the monolithic bed is used for the generation of the electroosmotic flow (EOF) from the anode to the cathode, but at the same time serves as a SCX stationary phase. A mixed‐mode (RP/SCX) mechanism for separation of peptides was observed in the monolithic column, comprising hydrophobic and electrostatic interaction as well as electrophoretic migration at a low pH value of mobile phase. A column efficiency of more than 280 000 plates/m for the unretained compound has been obtained on the prepared monoliths. The relative standard deviations observed for t 0 and retention factors of peptides were about 0.32% and less than 0.71% for ten consecutive runs, respectively. Effects of mobile phase compositions on the EOF of the monolithic column and on the separation of peptides were investigated. The selectivity on separation of peptides in the monolithic capillary column could be easily manipulated by varying the mobile phase composition.