A technique of High Performance Liquid Chromatography was applied to determine trehalose in the samples, which could contain other disaccharides (maltose, sucrose). Two columns and various mobile phases of eluents were tested as regards the efficiency of resolution. The limits of detection and determination were estimated as were the range of linearity and the precision of the selected methods. Moreover, the process of extracting trehalose from yeast cells was as regards the disintegrating substance, temperature, and extraction duration time).