The radionuclide 99 Mo has been prepared by 100 Mo(c, n) reaction using two types of natural molybdenum compound (molybdenum trioxide and zirconium molybdate gel) with the bremsstrahlung end-point energies of 10 and 15 MeV. After the equilibrium, 99m Tc was separated as NaTcO 4 from the irradiated samples using two different chemical procedures to examine the chemical yield. The separated Na[ 99m Tc]TcO 4 from the ZrMo gel has the 99Mo breakthrough of \10 -4 %, radiochemical purity[99 % as well as chemical impurities of Al, Mo and Zr \ 10 ppm. The estimation of 99 Mo and 99m Tc was done by using off-line c-ray spectrometric technique. The chemical yield of the separated 99m Tc from the dissolved molybdenum trioxide is 70.7-75.2 %, whereas in the undissolved zirconium molybdate gel, it is 19.1-43 %. The second method is preferable because it is user friendly in hospital radiopharmacy throughout the shelf-life.Keywords Medical isotope 99 Mo-9m Tc Á 100 Mo(c,n) reaction Á Molybdenum trioxide Á Zirconium molybdate gel Á Off-line c-ray spectrometric technique Á Chemical yield