Nano magnesium aluminum layered double hydroxide (nano MgAl LDH) sorbent, prepared via autocombustion route, was used as a base material for chromatographic 99Mo/ 99mTc column generator. The acid–base titration curve proved the amphoteric nature of the prepared MgAl LDH (with a point of zero charge at pH 6.5). The sorbent prepared with the optimum conditions was characterized by IR spectroscopy and XRD. The Mo(VI) breakthrough capacity was found to be 6.54´10-2 mmol Mo(VI)/g, i.e., 6.27 mg Mo(VI)/g. The prepared 99Mo/99mTc chromatographic column generator showed a good performance. The 99mTc elution yield was found to be 75.0 % at flow rate of 1 ml/min, pH of the eluate ranged from 5.5-7.5, the radionuclidic purity of the eluted 99mTc was > 99.99 % and Al3+ content in the eluate was < 5 ppm. In conclusion Nano magnesium aluminum layered double hydroxide (MgAl LDH) sorbent was well synthesized and characterized and used as a base material for 99Mo/99mTc column generator achieving higher loading capacities than that of alumina.