Surface charge is an important property of particles. It has been utilized to separate particles in microfluidic devices, where dielectrophoresis (DEP) is often the driving force. However, current DEP-based particle separations based on the charge differences work only for particles of similar sizes. They become less effective and may even fail for a mixture of particles differing in both charge and size. We demonstrate that our recently developed AC insulator-based dielectrophoresis (AC iDEP) technique can direct microparticles toward charge-dependent equilibrium positions in a ratchet microchannel. Such charge-based particle separation is controlled by the imposed AC voltage frequency and amplitude but is nearly unaffected by the size of either type of particle in the mixture except for the time required to achieve an effective separation. This AC iDEP technique may potentially be used to focus and separate submicron or even nanoparticles because of its virtually "infinite" channel length.