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Optical Engineering

Modern Aspects Of Fluorometric Detection In Liquid-Phase Chromatography
Author(s): Bernard Bousquet; Jean Pierre Garnier; Claude Dreux
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Paper Abstract

Recent advances are described in the combined use of fluorometric derivatization and high performance liquid chromatography (HPLC) for clinical chemistry determinations. Derivatives (especially dansyl derivatives) can be formed prior to chromatography in the case of estrogens, amino acids, and catecholamines. In post-column reactions, we preferred to use air-segmented reactions as they conform better to all the optimized chromatographic and spectrofluorometric parameters. Fluorescent derivatives produced from cate-cholamines, tryptophan and its metabolites, hydroxyindoles, tryptamine, amino acids, sugars, polyamines, and other substances are often sufficiently sensitive to be detected in picogram quantities by HPLC. Their reaction principle and some of their applications to samples are described. Recently, chemical excitation of fluorophore-like dansyl amino acid was proposed as a detection system for HPLC. By a post-column reaction, a fluorophore can be made to emit light by its reaction with trichlorophenyl oxalate (TCPO) and hydrogen peroxide. The detection limit of this system is about 10 fmol for each dansyl amino acid. Application of this new reaction to catecholamines opens up new prospects for fluorometric detection.

Paper Details

Date Published: 1 October 1983
PDF: 3 pages
Opt. Eng. 22(5) 225589 doi: 10.1117/12.7973201
Published in: Optical Engineering Volume 22, Issue 5
Show Author Affiliations
Bernard Bousquet, Hopital Saint-Louis (France)
Jean Pierre Garnier, Hopital Saint-Louis (France)
Claude Dreux, Hopital Saint-Louis (France)

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