The measurement of catecholamines and their metabolites in biological samples remains a current analytical challenge, in spite of the great diversity of methodologies that have been developed throughout the years. High-performance liquid chromatography is the standard method for their separation and quantification in biological samples, either coupled with electrochemical, fluorescence, chemiluminescence or mass spectrometry detection. This review summarizes the most important physicochemical properties of catecholamines, the wide panoply of sample preparation techniques and the main issues to consider during the development of chromatographic methods. The major difficulties encountered during the optimization of these procedures are related with the high tendency of catecholamines to oxidize and the very low quantities at which they exist in biological matrices. Herein, the most important aspects that ought to be considered during collection, treatment and storage of fluid and tissue samples intended for catecholamine analysis are underlined, the chromatographic conditions are compared and the technical advantages and limitations of each detection system are discussed.
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