Fluorescence Detection in High Performance Liquid Chromatography

Abstract

The principles and applications of fluorescence detection and fluorescence introducing reagents and methods in HPLC are reviewed. The design and requirements for fluorescence detectors, flow cells and excitation sources and the conversion of non-fluorescent compounds into fluorescent products by pre-column and post-column derivatization reactions are discussed. For the applications the emphasis is on drug analysis, where possible in biological fluids (serum, urine, etc.). The last paragraphs are divided in a number of sections in which newly developed and some scarcely used reagents are mentioned shortly; a more complete treatment is given of the reagents and labels most frequently used in the derivatization of certain functional groups. In this discussion the methods of derivatization as well as the selectivity, stability, fluorescence behaviour of the reagents/labels and derivatives and the reaction conditions are included. An up-to-date survey of the applications of fluorescence detection in liquid chromatography (TABLE III, TABLE IV and TABLE V), ends this review paper.