3-Aminobenzamide and 3-aminobenzoic acid, tags for capillary electrophoresis of complex carbohydrates with laser-induced fluorescent detection.
The efficiencies in derivatization of reducing carbohydrates were compared by capillary electrophoresis using maltose as a model with nine monoaminobenzene derivatives by reductive amination in the presence of sodium cyanoborohydride. We found that aminobenzene derivatives substituted at the 3-position showed good reactivity with reducing carbohydrates as expected from the reaction mechanism, although the fluorescence intensities and molar absorptivities of these derivatives were not as high as those of 2- and 4-aminobenzene derivatives. The reagents, 3-aminobenzamide and 3-aminobenzoic acid, which showed the highest reactivity, were applied to the labeling of carbohydrate chains obtained from some sialic acid-containing glycoprotein samples, and also high-mannose and hybrid-type oligosaccharides. Capillary electrophoresis of these labeled carbohydrate chains in an inner surface-modified capillary with (50% phenyl)methylpolysiloxane allowed excellent separation of sialic acid-containing carbohydrate chains derived from fetuin and thyroglobulin as well as high mannose-type and hybrid-type carbohydrates derived from bovine pancreas ribonuclease B, soybean agglutinin and hen ovalbumin. The lower limit of calibration was as low as the 10(-16) mol (injected amount) with helium-cadmium laser induced detection.
Kakehi
K
K
Faculty of Pharmaceutical Sciences, Kinki University, Higashi-Osaka, Japan. k_kakehi@phar.kindai.ac.jp
Funakubo
T
T
Suzuki
S
S
Oda
Y
Y
Kitada
Y
Y
D062365:Aminobenzoates; D001549:Benzamides; D002241:Carbohydrates / Q000032:analysis*; D019075:Electrophoresis, Capillary / Q000379:methods*; D006023:Glycoproteins / Q000032:analysis; D007202:Indicators and Reagents; D007834:Lasers; D008358:Mannose / Q000032:analysis; D019158:N-Acetylneuraminic Acid / Q000032:analysis; D009844:Oligosaccharides / Q000032:analysis; D013050:Spectrometry, Fluorescence; D013954:Thyroglobulin / Q000032:analysis; D014168:Transferrin / Q000032:analysis; D000509:alpha-Fetoproteins / Q000032:analysis; D062368:meta-Aminobenzoates
D062365:Aminobenzoates; D001549:Benzamides; D002241:Carbohydrates; D006023:Glycoproteins; D007202:Indicators and Reagents; D009844:Oligosaccharides; D014168:Transferrin; D000509:alpha-Fetoproteins; D062368:meta-Aminobenzoates; C025160:3-aminobenzamide; D013954:Thyroglobulin; C044855:3-aminobenzoic acid; D019158:N-Acetylneuraminic Acid; D008358:Mannose