USE OF NATIVE FLUORESCENCE MEASUREMENTS AND STOPPED-FLOW SCANNING TECHNIQUE IN THE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC ANALYSIS OF CATECHOLAMINES AND RELATED COMPOUNDS

Simultaneous separation of catecholamines and typtophan metabolites has been carried out using a reversed-phase partition mode of high-performance liquid chromatography (HPLC). Compounds are detected and measured via their native fluorescence emitted with an excitation wavelength of 285 nm and the emission cut-off filter of 340 nm. Sample preparation is minimized and the assay is selective and well-suited for routine analyses. The sensitivity of the method is in the nanogram range. The identity of chromatographic peaks is confirmed by their excitation spectra, obtained by the stopped-flow fluorescence scanning method. This method is applied to the direct analysis of rat brain and heart extracts as well as human serum samples. We believe that the well known separation power of HPLC, combined with the current state of the art in fluorescence monitoring will circumvent some of the problems presently encountered in the analysis of these compounds of great biological importance. © 1979.