The applicability of palladium for the separation of hydrogen isotopes (hydrogen and deuterium) is evaluated systematically by generating isotherm data and conducting column experiments in a laboratory set-up. Effect of various parameters such as concentration of the isotopic mixture, particle size, eluent flow rate, etc. is studied experimentally. A fixed-bed chromatographic model is developed and validated using the experimental data. The model is further used to predict the performance of a multi-column configuration for large-scale separation. Chromatographic separation is thus found to be a promising technique to achieve the required purity and hence it may be clubbed with the existing systems (e.g. cryogenic distillation) to obtain enhanced performance.