High-performance liquid chromatographic (HPLC) separation of sugar anomers was carried out on -NH2columns in acidic eluents in a very low temperature region. Chromatogram patterns were found to show remarkable changes with decreasing column temperatures. This was attributed to the fact that interconversion between sugar anomers (mutarotation) occurred during the chromatographic process. Since mutarotation was suppressed in a very low temperature region, pyranose anomers of each mono- and di-saccharide were separated completely. Furanose anomers were also separated from an equilibrium mixture, and the necessary conditions for complete separation of furanose anomers were discussed.