Palladium is widely used in hydrogen isotope technology owing to its obvious isotope effect, the systematic research of protium-deuterium seperation factor of sponge palladium was conducted by quadrupole mass spectrometer. The impact of temperatre (-25 ℃~155 ℃), protium-deuterium relative abundance (5%~90%), hydrogen-palladium ratio in the solid phase (0.05~0.7) on protium-deuterium seperation factor of sponge palladium was analyzed by experimental method. Combining the calculation of that in the a phase, the results show that protium-deuterium seperation factor of sponge palladium decreases with temperature, and increases with deuterium relative abundance. Protium-deuterium seperation factor increases with hydrogen-palladium ratio in the a+β and β phase, but such effect was not observed in the α phase.