Capillary zone electrophoresis of malto-oligosaccharides derivatized with 8-aminonaphthalene-1,3,6-trisulfonic acid.
Malto-oligosaccharides were derivatized via their reducing end with 8-aminonaphthalene-1,3,6-trisulfonic acid by reductive amination, and the separation and electrophoretic migration behavior of the labelled sugars were investigated by capillary zone electrophoresis. Series of linear malto-oligosaccharides were found particularly suitable for both the study of the effect of the operating conditions on the separation and the investigation of the relationship between the electrophoretic mobility and the molecular size of the homologues. The electrophoretic mobility of the malto-oligosaccharide conjugates was found to be a linear function of the molecular mass to the negative two-thirds power. The sugar derivatives employed here carry three negative charges due to the presence of the dissociated sulfonic acid groups even at strongly acidic pH. Therefore, the analytes can migrate in the electric field without interference by electroendosmotic flow and/or wall adsorption in uncoated silica capillaries at low pH. As a result, the separation of these carbohydrate conjugates can be carried out under such conditions with high speed and efficiency in free solution, i.e., without an anticonvective medium such as a gel or a viscous polymer solution. Appropriate use of triethylammonium phosphate buffer, pH 2.5, as the background electrolyte improves not only the reproducibility, but also the efficiency and speed of the separation. The labelled sugars allow monitoring of the separation by UV detector or laser-induced fluorescence detector with concomitant enhancement of analytical sensitivity.
Chiesa
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Department of Chemical Engineering, Yale University, New Haven, CT 06520.
Horváth
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D002240:Carbohydrate Sequence; D004586:Electrophoresis; D004591:Electrophoresis, Polyacrylamide Gel; D005947:Glucose / Q000032:analysis; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007834:Lasers; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009281:Naphthalenes; D009844:Oligosaccharides / Q000302:isolation & purification*; D009995:Osmosis; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D013213:Starch / Q000032:analysis; D013757:Tetraethylammonium Compounds
D009281:Naphthalenes; D009844:Oligosaccharides; D013757:Tetraethylammonium Compounds; D013213:Starch; C046465:8-amino-1,3,6-naphthalenetrisulfonic acid; D005947:Glucose