Experimental study for hydrogen isotope separation by thermal diffusion has been performed by using a single column and four interlinked columns. The dependence of the separation factor Q on the pressure was examined for single column operation with H-D (H2/ D2 ≈ 52/48), H-T (H2 / T2 ≈ 50/50) and D-T (D2 / T2 ≈ 80/20) systems where the hot wire temperature selected was 1273K. The optimum pressure observed was 93kPa for H-T, and 106kPa for H-D and D-T systems. The four columns were interlinked to function as a single column under the total reflux mode, and were operated with H-T at the optimum pressure observed in the single column experiments. The composition distributions experimentally observed were in close agreement with calculated results of the single column operation which is four times as high as each column.