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On‐line affinity selection of histidine‐containing peptides using a polymeric monolithic support for capillary electrophoresis
Author(s) -
Vizioli Nora M.,
Rusell Maria Lucía,
Carbajal Maria Laura,
Carducci Clyde N.,
Grasselli Mariano
Publication year - 2005
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.200410416
Subject(s) - iminodiacetic acid , capillary electrophoresis , chromatography , glycidyl methacrylate , monolithic hplc column , capillary action , chemistry , electrophoresis , histidine , self healing hydrogels , imidazole , methacrylate , analytical chemistry (journal) , polymerization , materials science , chelation , polymer , high performance liquid chromatography , polymer chemistry , amino acid , biochemistry , organic chemistry , composite material
An on‐line affinity selection method using a polymeric monolithic support is proposed for the retention of histidine‐containing peptides and their subsequent separation by capillary zone electrophoresis (CZE). Monolithic capillary columns were prepared in fused‐silica capillaries of 150 μm inner diameter (ID) by ionizing radiation‐initiated in situ polymerization and cross‐linking of diethylene glycol dimethacrylate and glycidyl methacrylate, and chemically modified with iminodiacetic acid (IDA) and copper ion. Monolithic microextractors were coupled on‐line near the inlet of the separation capillary (fused‐silica capillary, 75 μm ID × 28 cm from the microextractor to the detector). Model peptide mixtures of histidine‐containing and histidine‐noncontaining peptides were assessed. Peptides were released from the sorbent by a 5 m M imidazole solution and then separated by CZE with ultraviolet detection. Relative standard deviation values for migration times and corrected peak areas were found to be lower than 5.8 and 10.5%, respectively. IDA‐Cu(II) ion modified monolithic microextractors showed a chromatographic behavior and could be reused at least 25 times. The use of monolithic supports proved to be an advantageous alternative to packed particles for the preparation of microextractors.