Diffusion as major source of band broadening in field-amplified sample stacking under negligible electroosmotic flow velocity conditions.

Theory of field-amplified sample stacking (FASS) also called field-enhanced sample stacking is reevaluated considering the early work of Chien, Burgi and Helmer. The classical theory presented by Chien, Helmer and Burgi predicts the existence of maxima, which are ascribed to the counteracting principles of zone focusing and hydrodynamic dispersion. In contrast to their work, we here focus on cationic analytes separated in an acidic background electrolyte providing a very low electroosmotic flow velocity. Therefore, peak broadening due to differences in the local electroosmotic flow velocities in different compartments of the capillary can be regarded to be negligible. Consequently, peak broadening resulting from hydrodynamic dispersion will not be the dominant limitation of the accessible enrichment efficiency. In our experimental studies we, however, obtain an optimum value for the field enhancement factor (maximum of the enrichment efficiency, when varying the electric conductivity of the sample and the size of the sample injection plug) corresponding to a 10-fold dilution of the BGE in the sample solution. Comparing these experimental data with data modeled according to the revised theory, we show that this limitation of the loadability is caused by the unavoidable decrease of the analyte migration velocity in the BGE compartment of the capillary when injecting of a sample plug of lower electric conductivity (decrease in the local electric field strength). The additional diffusional band broadening limits the obtainable enrichment efficiency.