Separation of pure fructo-oligosaccharides from the mixed solution of glucose, sucrose and fructo-oligosaccharides was studied using a cationic ion-exchange resin column. Optimum separation conditions, i.e., temperature, feeding rate and the ratio of column vs. diameter were evaluated, which were found to be $85^{circ}C$, $0.25h^{-1}$ and 30, respectively. At the optimized separation conditions, high-purity fructo-oligosaccharides up to 96% were obtained and the total recovery yield was about 66% after four cycles. After the chromatographic separation, purification to remove the salts and color in pure fructo-oligosaccharides solution was successfully conducted using the mixed-bed of cationic and anionic ionexchange resin columns.