Determination of aliphatic thiols by fluorometric high-performance liquid chromatography after precolumn derivatization with 2-(4-N-maleimidophenyl)-6-methylbenzothiazole.
A sensitive, fluorometric high-performance liquid chromatographic method for the detection of aliphatic thiols, following pre-column derivatization with 2-(4-N-maleimidophenyl)-6-methylbenzothiazole, has been developed. The N-maleimide, the acid and the methyl ester derivatives of the commercially available 2-(4-aminophenyl)-6-methylbenzothiazole were synthesized and found to be equally effective for the precolumn derivatization procedure. The resulting fluorescentic derivatives of aliphatic thiols were separated on a reversed-phase column (Ultrasphere-ODS) using 0.1% hexane-sulfonic acid in 10 mM potassium hydrogen phosphate: acetonitrile (65:35) as a mobile phase and were detected fluorometrically (excitation 320 nm; emission 405 nm). The method is highly sensitive (femtomole range) and is easily applied for determination of SH-containing drugs and endogenous thiols in biological samples.
Haj-Yehia
A I
AI
Department of Pharmacy, School of Pharmacy, University of California, San Francisco 94143-0446, USA.
Benet
L Z
LZ
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003545:Cysteine / Q000032:analysis / Q000493:pharmacokinetics; D005470:Fluorometry / Q000379:methods; D005978:Glutathione / Q000032:analysis / Q000493:pharmacokinetics; D007202:Indicators and Reagents / Q000138:chemical synthesis*; D008297:Male; D008301:Maleimides / Q000138:chemical synthesis*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012680:Sensitivity and Specificity; D013438:Sulfhydryl Compounds / Q000032:analysis* / Q000493:pharmacokinetics; D013844:Thiazoles / Q000138:chemical synthesis*; D014018:Tissue Distribution
C093186:2-(4-N-maleimidophenyl)-6-methylbenzothiazole; D007202:Indicators and Reagents; D008301:Maleimides; D013438:Sulfhydryl Compounds; D013844:Thiazoles; D005978:Glutathione; D003545:Cysteine