Summary
A new system for the rapid and sensitive analysis of underivatized carbohydrates has been established using capillary zone electrophoresis with indirect UV detection. At an applied potential of 28 kV, sugars and sugar acids could be separated by the combined effects of electroendosmosis and electrophoresis within 20 minutes in a fused silica capillary of 50 μm internal diameter and an effective length of 100 cm using 6mM sorbic acid, pH 12.1, as both carrier electrolytie and chromophore. The alkaline pH ensured ionization of the sugars and, hence, their detection by means of charge displacement. Furthermore, the chosen concentration of sorbic acid allowed the smallest fractional change in the background signal to be measured. While the electrophoretic mobilities of the sugars were found to increase within a pH range of 11.9 to 12.3, those of the sugar acids were not affected. Due to the increasing competition of hydroxide ions in the displacement of the chromophore with rising pH, a significant loss of sensitivity is observed at pH values higher than 12.1 and this pH was found to provide sufficient resolution, optimum sensitivity, and a acceptably short analysis time. Under these conditions, a lower detection limit of 2 pmol was obtained for glucose.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
E. Stahl, U. Kaltenbach, J. Chromatogr.,5, 351 (1961).
R.A. Laine, W.J. Esselman, C.C. Sweeley, Methods Enzymol.,28, 159 (1972).
S.C. Churms, J. Chromatogr.,500, 555 (1990).
S. Hoffstener-Kuhn, A. Paulus, E. Gasmann, H.M. Widmer, Anal. Chem.,63, 1541 (1991).
J. Böseken, Adv. Carbohydr. Chem.,4, 189 (1949).
H. Weigel, Adv. Carbohydr. Chem.,18, 61 (1963).
H. Scherz, Electrophoresis,11, 18 (1990).
S. Hase, S. Hara, Y. Matsushima, J. Biochem.,85, 217 (1979).
S. Honda, S. Iwase, A. Makino, S. Fujiwara, Anal. Biochem.,176, 72 (1989).
W. Nashabeh, Z. El Rassi, J. Chromatogr.,514, 57 (1990).
T.W. Garner, E.S. Yeung, J. Chromatogr.,515, 639 (1990).
P. Jandik, W.R. Jones, J. Chromatogr.,546, 431 (1991).
F. Foret, S. Fanali, L. Ossicini, P. Bocek, J. Chromatogr.,470, 299 (1989).
J.A. Rendle, Jr., in “Carbohydrates in Solution”,H.S. Isbell, ed., American Chemical Society, Washington, DC, 1973, p. 54.
T. Tsuda, K. Nomura, G. Nakagawa, J. Chromatogr.,248, 241 (1982).
G. Bonn, H. Binder, H. Leonhard, O. Bobleter, Monatsh. Chem.,116, 961 (1985).
E.S. Yeung, Acc. Chem. Res.,22, 125 (1989).
R. Pecina, G. Bonn, E. Burtscher, O. Bobleter, J. Chromatogr.,287, 245 (1984).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Vorndran, A.E., Oefner, P.J., Scherz, H. et al. Indirect UV detection of carbohydrates in capillary zone electrophoresis. Chromatographia 33, 163–168 (1992). https://doi.org/10.1007/BF02275899
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02275899