On-column derivatization for the analysis of homocysteine and other thiols by capillary electrophoresis with laser-induced fluorescence detection

On-column derivatization and capillary electrophoresis (CE) with laser-induced fluorescence (LIF) detection has been developed for the fully automated assay of homocysteine and other thiols. The unique feature of this CE technique comes from the direct injection of a sample including homocysteine, enabling the derivatization with 4-aminosulfonyl-7-fluoro-2,1,3-benzoxadizole (ABD-F) to be accomplished in the capillary. After the derivatization for 10 min at 50°C, the homocysteine was analyzed within 7 min under an applied electric field of 333 V cm−1. The detection limit obtained for homocysteine with on-column LIF detection was 5.0 nM, as compared to 2.5 nM with pre-column LIF detection. The method is a very simple, fast, and practical approach for the fully automated assay of homocysteine and other thiols contained in low-volume and low-concentration samples.