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 degrees 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.
Kang
S H
SH
Department of Chemistry, Iowa State University, Ames 50011, USA.
Wei
W
W
Yeung
E S
ES
D019075:Electrophoresis, Capillary / Q000379:methods*; D006710:Homocysteine / Q000032:analysis*; D007834:Lasers; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods; D013056:Spectrophotometry, Ultraviolet / Q000379:methods; D013438:Sulfhydryl Compounds / Q000032:analysis*
D013438:Sulfhydryl Compounds; D006710:Homocysteine