Abstract: The separation and recovery of hydrogen-helium and tritium-helium mixture were a key technology for the tritium production process. A separation experiment device for hydrogen-helium and tritium-helium mixture was made by the low temperature adsorption desorption method with the double towers fixed bed, using the difference of adsorption capacity for adsorbents between hydrogen isotopes and helium at liquid nitrogen. The separation experiment of hydrogen-helium and tritium-helium mixture with different composition was developed on this apparatus. The results show that hydrogen-helium and tritium-helium mixture with different composition are quickly separated by the low temperature adsorption-desorption method with the double towers fixed bed, and the hydrogen content in helium is below 1.2×10^-5, and helium purity reach 99.998 8%, and detritiation factors of the adsorption column is more than 97%. The low temperature adsorption-desorption method with the double towers is not suitable for the hydrogen recovery, because of desorption hydrogen containing high concentration helium. It is feasible for tritium-helium separation using the fixed bed low temperature adsorption column and acting auxiliary unit of membrane separator.