In this paper, we introduce a dielectrophoresis (DEP)-based separation method that allows for tunable multiplex separation of particles. In traditional DEP separations where the field is applied continuously, size-based separation of particles uses the cubic dependence of the DEP force on particle radius, causing large particles to be retained while small particles are released. Here we show that by pulsing the DEP force in time, we are able to reverse the order of separation (eluting the large particles while retaining the small ones), and even extract mid-size particles from a heterogeneous population in one step. The operation is reminiscent of prior dielectrophoretic ratchets which made use of DEP and Brownian motion, but we have applied the asymmetric forces in time rather than in a spatial arrangement of electrodes, thus simplifying the system. We present an analytical model to study the dynamic behavior of particles under pulsed DEP and to understand the different modes of separation. Results from the model and the experimental observations are shown to be in agreement.