2-(4-N-MALEIMIDOPHENYL)-6-METHOXYBENZOFURAN - A SUPERIOR DERIVATIZINGAGENT FOR FLUOROMETRIC-DETERMINATION OF ALIPHATIC THIOLS BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY
Ai. Hajyehia et Lz. Benet, 2-(4-N-MALEIMIDOPHENYL)-6-METHOXYBENZOFURAN - A SUPERIOR DERIVATIZINGAGENT FOR FLUOROMETRIC-DETERMINATION OF ALIPHATIC THIOLS BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY, Journal of chromatography B. Biomedical applications, 666(1), 1995, pp. 45-53
Citations number
23
Categorie Soggetti
Chemistry Analytical
Journal title
Journal of chromatography B. Biomedical applications
The use of 2-(4-N-maleimidophenyl)-6-methoxybenzofuran as a fluorogeni
c pre-column derivatizing agent for sensitive high-performance liquid
chromatographic (HPLC) determination of aliphatic thiols is described.
The maleamic acid, its methyl ester and the N-maleimide derivatives o
f the common amine precursor were easily synthesized and characterized
according to conventional methods. These derivatives lack fluorescent
properties in their native form but display strong fluorescence upon
reaction with aliphatic thiols. The reaction is rapid and highly selec
tive for thiols over a wide pH range (7.1-8.8). Following derivatizati
on, the thiol adducts were separated on a reversed-phase column (Ultra
sphere-ODS) using 0.1% hexanesulfonic acid in 10 mM potassium hydrogen
phosphate-acetonitrile (65:35, pH 4.5) and were detected fluorimetric
ally (excitation at 310 nm; emission a; 390 nm). The method is highly
sensitive (femtomole range) and is easily applied to the determination
of SH-containing drugs and endogenous thiols in biological samples.