Indirect detection by semiconductor laser-induced fluorometry electrokinetic chromatography

Indirect fluorescence detection of electrically neutral compounds separated by micellar electrokinetic chromatography is performed using a semiconductor laser as an exciting light source. Oxazine 750 is used as a visualizing agent of which absorption maximum is near 680 nm. A surfactant, tetradecyltrimethylammonium chloride, is used to form micelles and to prevent adsorption of oxazine 750 with a positive charge on the capillary wall negatively charged. This surfactant coats on the capillary wall so that oxazine 750 is repulsed electrically on the capillary wall. In this technique, some aromatic compounds with relatively polar functional groups, such as aniline and nitrobenzene, could be separated and detected, while nonpolar compounds such as benzene and toluene can not be detected. The range of the detection limit is from 4.2 × 10^-4 to 1.6 × 10^-3 M (S/N = 3) for the aromatic compounds. The detection mechanism is based on enhancement of the fluorescence intensity in the micellar solution and on exclusion of the fluorophore attached at the hydrophilic moiety of the micelle by a hydrophilic sample.