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Preparative capillary zone electrophoresis using a dynamic coated wide‐bore capillary
Author(s) -
Yassine Mahmoud M.,
Lucy Charles A.
Publication year - 2006
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.200500862
Subject(s) - capillary action , capillary electrophoresis , chemistry , chromatography , tris , electrophoresis , pulmonary surfactant , analytical chemistry (journal) , dispersion (optics) , cytochrome c , adsorption , hydroxymethyl , bromide , materials science , inorganic chemistry , biochemistry , physics , organic chemistry , stereochemistry , optics , mitochondrion , composite material
Preparative capillary zone electrophoresis separations of cytochrome c from bovine and horse heart are performed efficiently in a surfactant‐coated capillary. The surfactant, dimethylditetradecylammonium bromide (2C 14 DAB), effectively eliminated protein adsorption from the capillary surface, such that symmetrical peaks with efficiencies of 0.7 million plates/m were observed in 50‐µm id capillaries when low concentrations of protein were injected. At protein concentrations greater than 1 g/L, electromigration dispersion became the dominant source of band broadening and the peak shape distorted to triangular fronting. Matching of the mobility of the buffer co‐ion to that of the cytochrome c resulted in dramatic improvements in the efficiency and peak shape. Using 100 mM bis(2‐hydroxyethyl)imino‐tris(hydroxymethyl)methane phosphate buffer at pH 7.0 with a 100‐µm id capillary, the maximum sample loading capacity in a single run was 160 pmol (2.0 µg) of each protein.