Use of capillary electrophoresis with chemiluminescence detection for sensitive determination of homocysteine

Abstract

A sensitive capillary electrophoresis (CE) method with chemiluminescence (CL) detection was developed for the determination of homocysteine (HCys) in human plasma. In this work, N-(4-aminobutyl)-N-ethylisoluminol was used as tagging reagent to label the analyte for achieving high assay sensitivity. N-(4-Aminobutyl)-N-ethylisoluminol-tagged HCys after CE separation reacted with hydrogen peroxide in the presence of horseradish peroxidase, producing CL emission. Experimental conditions for labeling analyte, CE separation, and CL detection were studied. The CL intensity was proportional to the concentration of HCys in the range of 2.5×10(-8) to 5.0×10(-6) M. Detection limit (S/N=3) was 7.6×10(-9) M. Human plasma samples from healthy donors were analyzed by the presented method. HCys levels were found to be in the range of 9.50-15.3 μM.

Simultaneous determination of total homocysteine and cysteine in human plasma by capillary zone electrophoresis with pH-mediated sample stacking

“…To elucidate the function of Hcy and Cys in biochemical processes and clinical practice, their identication and accurate measurement are essential. In recent years, several CE procedures have been applied [7][8][9][10][11][12] to evaluate Hcy and thiols in general, in body uids. Many electrophoretic procedures for thiols and similar or related compounds have employed LIF 5,[13][14][15][16][17][18][19][20] detection but also UV [21][22][23][24][25][26] and amperometric [27][28][29][30][31] detection have been proposed with success.…”

Preparation of a dual‐enzyme co‐immobilized capillary microreactor and simultaneous screening of multiple enzyme inhibitors by capillary electrophoresis

“…CE is a powerful separation tool with the advantage of high‐efficiency separation, short analysis times and low sample loading , and studies have shown that CE is a versatile platform for enzyme study and inhibitor screening . Chenet et al developed a CE method for the screening of 20 proteasome inhibitors .…”

Recent advances in chemiluminescence detection coupled with capillary electrophoresis and microchip capillary electrophoresis

“…CE–CL detection method has also been applied to amino acid , PCR product , saccharide , organic small molecule , and metal ion analysis.…”