CHROMATOGRAPHY, Vol. 22 (2001), No. 3, pp. 171-180

Analysis of Carbohydrates by Capillary Electrochromatography
Shigeo Suzuki and Susumu Honda*
Faculty of Pharmaceutical Sciences, Kinki University, 3-4-1, Kowakae, Higashiosaka, Japan

In this review we summarize the recent advances in capillary electrochromatography of carbohydrates, glycoconjugates, and carbohydraterelated compounds. Intact carbohydrates could be separated on ODS columns with aqueous acetonitrile as eluent. However, detection by UV absorption in the ordinary wavelengths did not give enough sensitivity for the detection of separated carbohydrates. Absorption at 195 nm allowed much more sensitive detection, but selectivity was rather problematic. Although the use of a light scattering detector was also possible, it had a similar limitation. Derivatization of carbohydrates enhanced detection sensitivity. Derivatives formed with 1-phenyl-3-methyl-5- pyrazolone could be separated on ODS columns with aqueous acetonitrile and sensitively detected by UV absorption. p-Nitrophenyl glycosides of mono- and oligosaccharides were also separated by this system. It was essential that a considerable proportion of silanol groups remained unmodified in the stationary phase in order to induce appropriate velocity of electroosmotic flow for driving the separated solutes inreasonable analysis time. However, the remaining silanol groups caused peak broadening. Tailor-made continuous bed of polyacrylamide columns gave excellent separation, since they were free from this “remaining silanol” problem. Some applications to glycoconjugates are also overviewed to evaluate CEC in biomedical analysis.
Keywords: capillary electrochromatography, high-performance liquid chromatography, carbohydrates, ODS columns, aminosilica columns, electroosmotic flow, in-capillary derivatization, PMP sugars, p-nitrophenylgylcosides

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