XXXX analysis of glycoconjugates by anion exchange chromatography with pulsed amperometric detection.

@entity1 @entity6146 @entity19131 @entity3363 @entity3954 @entity7787 @entity11485 @entity9928 @entity4823 @entity3953 @entity3913 @entity6674 @entity3649 @entity12016 @entity413
The method of anion exchange chromatography followed by pulsed amperometric detection (AE-PAD; Johnson, D. C., and Polta, T. Z. (1986) Chromatogr. Forum 1, 37-44) has been applied to the compositional analysis of glycoconjugates. Using 22 mM @entity6674 as a column effluent, underivatized @entity3953 , @entity11485 , @entity4823 , @entity3649 , @entity413 , and @entity3913 were readily separated in 15 min at a flow rate of 1 ml/min. The limit of quantification of the @entity7787 was better than 100 pmol (signal to noise ratio 184:1). AE-PAD was employed to quantify the @entity7787 of several glycoproteins, glycopeptides, and oligosaccharides after hydrolysis with 2 M @entity6146 . Both neutral and @entity12016 could be rapidly estimated in a single chromatographic step using AE-PAD. Complete release of @entity3954 required more vigorous hydrolysis conditions (Lee, Y. C. (1972) in Methods in Enzymology (Ginsburg, V., Ed.), Vol. 28, pp. 63-73, Academic Press, New York). In both glycopeptides and oligosaccharides, approximately one less residue of @entity1 than predicted was determined. Both AE-PAD and liquid chromatographic analysis of @entity19131 - @entity9928 complexes with fluorometric detection (Mikami, H., and Ishida, Y. (1983) Bunseki Kagaku 32, E207-E210) gave similar quantification of @entity3913 and other @entity3363 . The capability of rapid, sensitive quantification of underivitized @entity7787 should facilitate structural analysis of glycoconjugates.
