DETERMINATION OF ALIPHATIC THIOLS BY FLUOROMETRIC HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY AFTER PRECOLUMN DERIVATIZATION WITH 2-(4-N-MALEIMIDOPHENYL)-6-METHYLBENZOTHIAZOLE
Ai. Hajyehia et Lz. Benet, DETERMINATION OF ALIPHATIC THIOLS BY FLUOROMETRIC HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY AFTER PRECOLUMN DERIVATIZATION WITH 2-(4-N-MALEIMIDOPHENYL)-6-METHYLBENZOTHIAZOLE, Pharmaceutical research, 12(1), 1995, pp. 155-160
A sensitive, fluorometric high-performance liquid chromatographic meth
od for the detection of aliphatic thiols, following pre-column derivat
ization with 2-(4-N-maleimidophenyl)-6-methylbenzothiazole, has been d
eveloped. The N-maleimide, the acid and the methyl ester derivatives o
f the commercially available 2-(4-aminophenyl)-6-methylbenzothiazole w
ere synthesized and found to be equally effective for the precolumn de
rivatization procedure. The resulting fluorescentic derivatives of ali
phatic thiols were separated on a reversed-phase column (Ultrasphere-O
DS) using 0.1% hexane-sulfonic acid in 10 mM potassium hydrogen phosph
ate: acetonitrile (65:35) as a mobile phase and were detected fluorome
trically (excitation 320 nm; emission 405 nm). The method is highly se
nsitive (femtomole range) and is easily applied for determination of S
H-containing drugs and endogenous thiols in biological samples.