Size-dependent electrophoretic migration and separation of liposomes by capillary zone electrophoresis in electrolyte solutions of various ionic strengths

The size-dependent electrophoretic migration and separation of liposomes was demonstrated and studied in capillary zone electrophoresis (CZE). The liposomes were extruded and nonextruded preparations consisting of phosphatidylcholine/phosphatidylglycerol/cholesterol in various ratios and ranging from 125 to 488 nm in mean diameter. When liposomes of identical surface charge density were subjected to CZE in Tris-HCl (pH 8) buffers of various ionic strengths (0.001-0.027), they migrated in order of their size. Size-dependent electrophoretic migration and separation of liposomes in CZE can be enhanced or brought about by decreasing the ionic strength of the buffer. It was shown that size-dependent migration is primarily a function of κR, where κ^-1 is the thickness of the electric double layer (which can be derived from the ionic strength, I, of the buffer) and R, the liposome radius. Liposome mobility depends on κR and surface charge density in a manner consistent with that expected from the Overbeek-Booth electrokinetic theory. Thus, the relaxation effect appears to be the physical mechanism underlying the size-dependent electrophoretic separation of liposomes.