Determination of thiols of biological and pharmacological interest by high-performance thin-layer chromatography and fluorescence scanning densitometry.

The application of high-performance thin-layer chromatography (HPTLC) with fluorescence scanning densitometry provides a simple, rapid and reliable system for the qualitative and quantitative determination of several thiols of biological and pharmacological interest. The determination of a mixture of thiols (captopril, coenzyme A, cysteamine, cysteine and glutathione), together with their disulphides may readily be performed by pre-chromatographic derivatization with the thiol-specific fluorobenzoxadiazole reagents SBD-F and ABD-F, followed by HPTLC separation on silica gel plates using isopropyl ether-methanol-water-acetic acid (9:8:2:1, v/v/v/v) as the developing solvent, and fluorodensitometric measurement of the fluorescing derivatives. Detection limits of about 30 pg (coenzyme A) to 6 pg (cysteamine) per spot were achieved; the relative standard deviation (RSD) of the complete procedure was 1.16-3.2%.