Molybdenum is a vital element in the industry and biological systems, and for environmental protection, it is needed to remove molybdenum ion from the wastewater. Therefore, this work focused on removal and separation of molybdenum ion from some fission products such as La(III), Eu(III), Ce(IV) and Y(III) ions. For this purpose, perlite was impregnated by trioctyl amine, characterized using XRF, SEM, FT-IR and XRD and used for the sorption and separation processes. Sorption behavior of these ions was investigated by the batch method. The impregnation process of perlite enhanced the percent uptake of Mo(VI) from 3.6 to 93.2%. Furthermore, separation tests using simulated waste solutions containing the mentioned fission products were carried out to verify the possibility of the separation process. The breakthrough curve of Mo(VI) had S-shaped profile, and the elution percentage was estimated to be 98.6% by 1.0 mol/L H2SO4. The obtained results revealed that the proposed process is essentially feasible.