SummaryMicellar electrokinetic capillary chromatography (MEKC), a mode of capillary electrophoresis, can separate cations, anions and uncharged molecules simultaneously and with high resolution. So it is ideal for separating endogenous and exogenous analytes in complex biofluids, such as urine. Our previous system used 13-CD-enhanced SDS MEKC to aid resolution and could resolve a large numberof peaks from human urine in about 12 minutes.In this study we have investigated a number of alternative additives to enhance resolution even further. Various compounds have been claimed, in the literature, to improve MEKC e.g. mixed micelles, organic solvents, and urea. We investigated a number of such compounds as additives in ourstandard urine system butwithout much success. However some modified cyclodextrins were found to increase both resolution and the migration window. In particular sulphated 13-cyclodextrin gave excellent improvements in both resolution and selectivif,/. The optimised conditions (6.2 mM S-13-CD in 25 mM sodium tetraborate, pH 10, 75 mM SDS) resolved 70 components in a normal urine pool in < 15 minutes using < 10 nl of urine. Faster separations butwith some loss of resolution were obtained by injection from the detector end of the capillary (short-end injection).In addition we have investigated the use of microemulsion electrokinetic chromatography MEEKC but found that it offered no real advantages over MEKC for urine analysis.This optimised MEKC separation is simpler, faster and gives higher resolution than gradient RP-LC.
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