4-(6,7-dihydro-5,8-dioxothiazolo[4,5-g]phthalazin-2-yl)benzoic acid N-hydroxysuccinimide ester as a highly sensitive chemiluminescence derivatizationreagent for amines in liquid chromatography

4-(6,7-Dihydro-5,8-dioxothiazolo[4,5-g]phthalazin-2-yl)benzoic acid N-hydro xysuccinimide ester was synthesized as a highly sensitive and selective che miluminescence derivatization reagent for primary and secondary amines in l iquid chromatography. Methyl-n-octylamine, n-nonylamine and n-decylamine we re used as model compounds to optimize the derivatization, separation and c hemiluminescence reaction conditions. This reagent reacts selectively with amines in the presence of triethylamine to give the highly chemiluminescent derivatives, which produce chemiluminescence by reaction with hydrogen per oxide in the presence of potassium hexacyanoferrate(III) in an alkaline med ium. The chemiluminescent derivatives of the three amines can be separated within 20 min by reversed-phase liquid chromatography with isocratic elutio n, followed by chemiluminescence detection. The detection limits (signal-to -noise ratio=3) for primary and secondary amines are at sub-fmol levels for a 20-mul injection. Furthermore, this method was applicable to the determi nation of amantadine in human plasma. (C) 2001 Elsevier Science B.V. All ri ghts reserved.