Separation of fructose and glucose was performed using emulsion liquid membranes with a mixture of an organoboronic acid and a quaternary ammonium salt as a carrier in a batch reactor. In order to find a carrier and an optimal experimental condition suitable to the sugar separation, extraction of each sugar was carried out independently. The effect of various experimental variables, such as initial concentration of sugar in the feed phase, type of organoboronic acids, and w/o ratio, on the sugar separation was investigated, and the concentrations of sugars in each aqueous phase were analyzed. The ratio of degree of extraction of fructose to that of glucose was very high, but the concentration of fructose in the receiving phase was not too high. Therefore, a stronger stripping agent in the receiving phase was required for development of a practical ELM system suitable to the sugar separation.