Joule heating induced stream broadening in free-flow zone electrophoresis.

Abstract

The use of an electric field in free-flow zone electrophoresis (FFZE) automatically leads to Joule heating yielding a higher temperature at the center of the separation chamber relative to that around the channel walls. For small amounts of heat generated, this thermal effect introduces a variation in the equilibrium position of the analyte molecules due to the dependence of liquid viscosity and analyte diffusivity on temperature leading to a modification in the position of the analyte stream as well as the zone width. In this article, an analytic theory is presented to quantitate such effects of Joule heating on FFZE assays in the limit of small temperature differentials across the channel gap yielding a closed form expression for the stream position and zone variance under equilibrium conditions. A method-of-moments approach is employed to develop this analytic theory, which is further validated with numerical solutions of the governing equations. Interestingly, the noted analyses predict that Joule heating can drift the location of the analyte stream either way of its equilibrium position realized in the absence of any temperature rise in the system, and also tends to reduce zone dispersion. The extent of these modifications, however, is governed by the electric field induced temperature rise and three Péclet numbers evaluated based on the axial pressure-driven flow, transverse electroosmotic and electrophoretic solute velocities in the separation chamber. Monte Carlo simulations of the FFZE system further establish a time and a length scale over which the results from the analytic theory are valid.