Micellar electrokinetic chromatography in zero-electroosmotic flow environment.

Abstract

Micellar electrokinetic chromatography (MEKC) is conducted in polyacrylamide-coated capillaries under almost complete suppression of electroosmotic flow. The equations of migration and resolution for neutral solutes in this mode of MEKC operation are presented. The technique is termed reversed-flow MEKC (RF-MEKC) because, in contrast to MEKC in bare-silica capillaries (N-MEKC), solute migration order is reversed and solute migration time is inversely proportional to micelle concentration. This presents an advantage for the high-efficiency separation of extremely and moderately hydrophobic solutes in a short analysis time. Examples of the separation of polycyclic aromatic hydrocarbons, aflatoxins and dansylated-amino acids are presented using sodium dodecyl sulfate (SDS) surfactant. Polycyclic aromatic hydrocarbons are separated using a relatively low micelle concentration. The detection sensitivity for these compounds is enhanced in two ways. First, the peaks are sharp because of the short analysis time and the inertness of the column surface. Second, the fluorescence background and Joule's heating are minimal because of the low concentration of SDS and other additives needed to affect the separation. While N-MEKC is mainly conducted with basic buffers, RF-MEKC can be conducted in basic as well as acidic media as illustrated in the separation of 15 dansylated-amino acids at pH 4.2.