Calibration based on the "single-point calibration method", a simple exponential transformation of the response function of an evaporative light scattering detector was improved and applied to analysis of selected saccharides under hydrophilic interaction chromatography mode (a polar phase LiChrospher100 DIOL, mobile phase acetonitrile/water). The improved approach to the calibration procedure yielded a calibration curve with an excellent linearity (quality coefficient <5%). This quantitative evaluation of chromatograms of D-galactose suggested that not only anomers but even pyranose and furanose forms of the anomers could be resolved--the resulting calculations of abundance of the anomeric form strongly correlated with data from the literature obtained mostly by NMR studies (analogous results were also obtained for D-arabinose, D-glucose, and D-mannose). Because of the rapid separation (retention time less than 10 min), the observed correlation enabled to monitor anomeric conversion (mutarotation) of monosaccharides.

Department of Natural Drugs Faculty of Pharmacy University of Veterinary and Pharmaceutical Sciences Brno Czech Republic

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