Electrostatic traveling wave (ETW) methods have grown in popularity as a method to manipulate and separate fine particles. This is of interest to mineral processing and In-situ Space Resource Utilization. Here, we demonstrate that ETW can be used to separate particles by size. First, our numerical calculation and simulation results show the competing effect of DEP and Coulomb force on particle motion direction, which depends on the frequency, particle size and charge. Then we define the “crossover frequency” allowing prediction of operating parameters to separate particles. At the “crossover frequency” – a particle is equally likely to move forward or backward. The crossover frequency decreases as particle size increases, and the proportion of particles moving backward increases as frequency increases. Four size groups of ballotini particles (glass) were transported and collected at different wave field frequencies. Finally, we have demonstrated experimentally the separation of particles between 30–50 µm and 75–110 µm. Simulation and experiment results revealed the effects of frequency, particle size and charge on particle motions. And the model can be used to guide the design of separation systems.