Abstract In this study, numerical simulations were carried out to investigate the separation of the helium-argon gas mixture by thermal diffusion column. This research determined the significant parameters and their effects on the process performance. Effects of feed flow rate, cut ratio, and hot wire temperature in a 950 mm height column with an inner tube of 9.5 mm radius were examined through the simulation of the thermal diffusion column. For minimizing the number of simulations and obtaining the optimum operating conditions, response surface methodology (RSM) was used. Analysis of separative work unit (SWU) values as a target function for helium-argon separation clearly showed that the maximum amount of SWU in thermal diffusion column was achieved, when hot wire temperature increased as large as technically possible, and the feed rate and cut ratio were equal to 55 Standard Cubic Centimeters per Minute (SCCM) and 0.44, respectively. Finally, the SWU value in optimum conditions was compared with the experimental data. Results illustrated that the experimental data were in good agreement with simulation data with an accuracy of about 90%.