Use of enhancers in the HPTLC fluorescence analysis of thiols

Several thiols of biological and pharmacological interest, including glutathione, coenzyme A, acetylcysteine and captopril were derivatized with the fluorogenic reagents SBD-F and ABD-F and analysed by high-performance thin-layer chromatography (HPTLC)-fluorodensitometry on silica gel 60 plates, using isopropyl ether-methanol-water-acetic acid (9:8:2:1, v/v/v/v) as the developing solvent. The luminescence was considerably increased when several types of enhancers were applied as dipping reagents: Triton X-100, liquid paraffin and cyclodextrins; thus the detectability of the thiol fluorophores was improved. The influence of enhancer concentration, method of application, sample concentration, drying conditions and measuring time after plate dipping were investigated. The greatest enhancement was achieved using a 40% (v/v) solution of Triton X-100 in toluene as a dipping reagent for the determination of SBD-acetylcysteine; more than a 10-fold increase of the fluorescence signal was obtained, allowing low picogram detection limits.