Application of micro-scale liquid chromatography with fluorescence detection to the determination of thiols

Different thiol compounds of biological and pharmacological interest were separated on a packed reversed-phase fused-silica capillary column and determined with fluorescence detection. Conventional inexpensive liquid chromatographic equipment could be adapted for such purposes, providing a highly efficient analytical system. The different compounds were derivatized with the fluorogenic reagents SBD-F and ABD-F and chromatographed both isocratically and in the gradient mode in order to separate a series of thiols with widely varying polarities. Subsequently the derivatives were measured at lambda ex. = 380 nm and lambda em. = 510 nm. Application of the system to biological and pharmacological samples is suggested.