Open AccessA novel capillary electrophoretic method for protein determination using bicinchoninic acid-Cu<SUP>+</SUP>colorimetric reaction
Author(s) -
孟庆威,
郭磊,
申睿,
谢剑炜
Publication year - 2010
Publication title -
sepu/chinese journal of chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.171
H-Index - 19
eISSN - 1872-2059
pISSN - 1000-8713
DOI - 10.3724/sp.j.1123.2010.00682
Subject(s) - bicinchoninic acid assay , chemistry , ricin , capillary electrophoresis , boric acid , chromatography , transferrin , detection limit , electrophoresis , bradford protein assay , biochemistry , organic chemistry , toxin
A new rapid, sensitive method for protein determination using capillary electrophoresis and specific colorimetric reaction of bicinchoninic acid (BCA) was established, assisted by microwave incubation. With 60 mmol/L boric acid buffer (pH 9.5) and inclusion additive of beta-cyclodextrin, the complex of BCA-Cu+ and free BCA molecules were efficiently separated. The peak intensity of BCA-Cu+ was higher than those of native proteins about two orders of magnitude at a low wavelength of 200 nm. The linear ranges of this method were from 2 to 200 mg/L for transferrin, and 2 to 100 mg/L for ricin. The limits of detection for transferrin and ricin were 0.33 and 0.37 mg/L, respectively. This method was also successfully applied in the determination of some ricin samples in the First International Proficiency Test on the quantification of ricin.