A high-performance liquid chromatographic (HPLC) method was established for kinetic studies of the interconversion (mutarotation) between pyranose sugar anomers in solution. At very low temperature, HPLC chromatograms of each sugar indicated the concentrations of each anomer in the state prior to the HPLC analysis. Thus, when one of the pyranose anomers obtained by recrystallization was dissolved into water and was analyzed by HPLC repeatedly, gradual changes in the chromatogrphic patterns were observed. The equilibrium concentrations of each sugar anomer in different solvents were found to vary, which was also investigated by HPLC.