Separating nanoparticles by surface charge group using pH-controlled passivated gel electrophoresis

Because ionically stabilized colloids in aqueous dispersions have net surface charges that depend on pH, it is potentially possible to separate mixtures of nanospheres having identical radii, yet different types of stabilizing surface charge groups, efficiently using passivated gel electrophoresis (gel-EP). To demonstrate this, we separate a binary dispersion of polystyrene nanospheres that have nearly identical radii and surface group densities, yet different types of anionic stabilizing surface charge groups: sulfate and carboxylate. We achieve an efficient separation by adjusting the pH of the running buffer to lie between the pKa values of these charge groups, resulting in significantly different protonation and, consequently, different electrophoretic propagation velocities of the nanospheres. The measured steady-state propagation velocities of both types of anionic nanoparticles as a function of pH can be fit well by an equilibrium model of pH-dependent protonation of anionic surface charge groups. Thus, pH-controlled passivated gel-EP opens a route for separating similarly sized charged colloidal objects that are stabilized by a variety of different surface charge groups.