2006 Volume 55 Issue 4 Pages 213-221
A highly selective and sensitive method with a novel concept is introduced for the assay of biological substances. This method is based on an intramolecular excimer-forming fluorescence derivatization with a pyrene reagent, followed by reversed-phase liquid chromatography. Polyamines, polyphenols, and polycarboxylic acids, having two or more reactive functional groups in a molecule, were converted to the corresponding polypyrene-labeled derivatives by a reaction with an appropriate pyrene reagent. These derivatives afforded intramolecular excimer fluorescence (440∼520 nm), which can clearly be discriminated from the monomer (normal) fluorescence (360∼420 nm) emitted from pyrene reagents and monopyrene-labeled derivatives of monofunctional compounds coexisting in the samples. By intramolecular excimer fluorescence detection, highly selective and sensitive determinations of polyamines including histamine and basic amino acids, polyphenols, and polycarboxylic acids including acidic amino acids could be achieved. Furthermore, the methods were successfully applied to the determination of various biological and environmental substances in real samples, which required only a small amount of samples and a simple pretreatment.
