In this article, the effect and optimization of process parameters in a counter-flow vortex tube on temperature difference were investigated through the Taguchi method. The experiments were planned as per Taguchi's L27 orthogonal array with each experiment performed under different conditions of inlet pressure, nozzle number, and cold mass fraction. By means of analysis of variance and regression analysis, the effects of factors and their interactions on temperature difference were determined and modeled with a correlation coefficient of 93.5%. Accordingly, it was observed that temperature difference goes up with the increase in inlet pressure, and the cold mass fraction and decreases with the increase in nozzle number. In addition, the optimum settings of process parameters maximizing the temperature difference are an inlet pressure of 650 kPa, a nozzle number of 2, and a cold mass fraction of 0.7. Finally, confirmation tests verified that the Taguchi method was successful in the assessment of vortex tube parameters for temperature difference.