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Repeatedly usable immobilized pH gradient in a monolithic capillary column
Author(s) -
Yang Chun,
Zhu Guijie,
Zhang Lihua,
Zhang Weibing,
Zhang Yukui
Publication year - 2004
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/elps.200405916
Subject(s) - glycidyl methacrylate , monolithic hplc column , monolith , chromatography , glutaraldehyde , isoelectric focusing , capillary action , chemistry , immobilized ph gradient , aqueous solution , methacrylate , polymer , polymerization , materials science , high performance liquid chromatography , organic chemistry , composite material , catalysis , enzyme
Glycidyl methacrylate (GMA) and ethylene dimethacrylate (EDMA) were used to synthesize a monolithic capillary column containing reactive epoxy groups. Glutaraldehyde was introduced and linked to the monolith after a process of amination. An aqueous solution of commercial carrier ampholytes (CAs, Ampholine) was focused in such a polymer column. The primary amino groups of CAs reacted with glutaraldehyde along the capillary. CAs were immobilized at different positions in the column according to their isoelectric points (p I ), resulting in a monolithic immobilized pH gradient (M‐IPG). Isoelectric focusing (IEF) was performed without CAs in such an M‐IPG column. Due to the covalent attachment of the CAs this M‐IPG can be repeatedly used after its preparation. Good stability, linearity, and reproducibility were obtained.