Mechanistic insights derived from retardation and peak broadening of particles up to 200 nm in diameter in electrophoresis in semidilute polyacrylamide solutions

Rigid spherical particles in the size range of 5–200 nm diameter were subjected to capillary zone electrophoresis (CZE) in semidilute solutions of un‐cross‐linked polyacrylamide of M r 5, 7 and 18 × 10 6 (PA‐5, ‐7 and ‐18, respectively) of varying concentrations up to 1.6% and at field strengths varying from 68 to 270 V/cm. For all particles under study, the experimental Ferguson plots, log(mobility) vs . polymer concentration, permit a linear approximation. Their slope, the retardation coefficient K R = ∂log(mobility)/∂(concentration), for particles smaller than 30 nm in diameter increased with particle size in PA‐5 and −7 independently of electric field strength and polymer M r . The K R of particles of 30 nm in diameter or more was found to be independent of particle size at the lowest field strength used but to decrease with it at the higher values of field strength. The decrease was parallel but shifted to higher values of retardation when the polymer M r increased from 5 to 7 × 10 6 . With a decreasing ratio of average mesh size of the polymer network, ξ, to particle radius, R , the approach to “continuity” of the polymeric medium (ξ/ R < 1) with both increasing particle size and polymer concentration does not result in the retardation behavior expected according to the macroscopic (bulk) viscosity of the solution. These experimental observations were hypothetically interpreted in terms of a transition to a retardation mechanism comprising the formation of a polymer depletion layer near the particle surface ‐ polymer solution interface. Peak width exhibited an overall increase with PA‐7 concentration for all particles studied. For particles of 30 nm in diameter or less, the increase was steepest when the radius of the particle was approximately commensurate with ξ at a given polymer concentration. For the largest particle, 205 nm in diameter, peak broadening with polymer concentration was found to correlate linearly with peak asymmetry. CZE of the particles in PA‐18 solutions revealed abnormal behavior, with both mobility and peak width remaining near‐constant up to a concentration of 0.08% and sharply declining at higher concentrations. The decline of relative mobility is the same for the entire particle size range used, while peak width declines in direct relation to particle size.