Influence of organic modifier concentration on plate number in micellar electrokinetic chromatography. 1. 2-propanol.

This paper establishes a physicochemical basis for the efficiency losses in micellar electrokinetic chromatography in buffers containing sodium dodecyl sulfate (SDS) and 2-propanol (2PN). Weakly, intermediately, and strongly retained analytes were separated in phosphate/borate buffers containing 50 mM SDS and from 0 to 10% 2PN by volume. Their plate numbers N generally agreed well with predictions of a theory for N based on longitudinal diffusion and instrumental contributions to dispersion. The N's of weakly and intermediately retained analytes were not affected strongly by 2PN over this concentration range, because their diffusion coefficients varied inversely with buffer viscosity and their retention times largely varied directly with viscosity. These combined effects on dispersion almost canceled. However, the N's of strongly retained analytes decreased with increasing 2PN, because their diffusion coefficients varied inversely with viscosity but their retention times increased more rapidly than did viscosity. These combined effects on dispersion did not cancel. These differences occurred because 2PN penetrated the micelles, caused bound counterions to be released, and increased the micellar charge and electrophoretic mobility. As 2PN concentration increased, the micelles electrophoresced increasingly rapidly against the electroosmotic flow. Consequently, strongly retained compounds required increasingly long times to elute.