On-line concentration of neutral analytes for micellar electrokinetic chromatography. 3. Stacking with reverse migrating micelles

On-line concentration of neutral analytes by sample stacking in reversed migration micellar electrokinetic chromatography is presented, Micellar separation solutions of sodium dodecyl sulfate are prepared with acidic buffers to reverse the direction of the migration velocity of neutral analytes owing to a reduced electroosmotic now. Samples are prepared in nonmicellar matrixes of low conductivity (i.e., water, diluted buffer, or dilute organic/aqueous solvent) to achieve field enhancement in the sample zone, Without polarity switching inherent in large-volume sample stacking, narrowing of analyte bands, removal of sample matrix, and separation of focused analyte bands are achieved. A model is proposed to describe the stacking technique and is supported by experimental results. In addition, equations are derived to describe: band broadening associated with the technique. Detector response improvements reaching a 100-fold are confirmed experimentally. Concentration detection limits on the order of low-ppb levels (S/N = 3) are realized with model steroidal compounds.