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%.
Ling
B L
BL
Laboratory of Pharmaceutical Chemistry and Drug Analysis, Faculty of Pharmaceutical Sciences, State University of Ghent, Belgium.
Baeyens
W R
WR
Del Castillo
B
B
Imai
K
K
De Moerloose
P
P
Stragier
K
K
D000085:Acetates / Q000032:analysis; D002855:Chromatography, Thin Layer; D003720:Densitometry; D004220:Disulfides / Q000032:analysis; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D013050:Spectrometry, Fluorescence; D013438:Sulfhydryl Compounds / Q000032:analysis*; D014867:Water / Q000032:analysis
D000085:Acetates; D004220:Disulfides; D007202:Indicators and Reagents; D013438:Sulfhydryl Compounds; D014867:Water