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Determination of the catecholamines and serotonin, their precursors tyrosine and tryptophan, and their main metabolites in rat brain using reversed-phase high-performance liquid chromatography with fluorimetric and oxidative amperometric detection in series

1992; Elsevier BV; Volume: 578; Issue: 1 Linguagem: Inglês

10.1016/0378-4347(92)80220-k

1872-812X

J.R. Lee Chin,

Neuroscience and Neuropharmacology Research

A high-performance liquid chromatographic method for the determination of catecholamines and serotonin, their precursors and their main metabolites was developed applied to rat cerebellum, hypothalamus, striatum and cortex. A fluorimetric and an oxidative amperometric detector were used in series. For both detectors, detection limits (25-520 pg) were useful for this application, linearity of standards was excellent (average r greater than 0.9997), between-run precision for sample analytes was generally acceptable (coefficient of variation less than 10% with appreciable concentrations present) and average recoveries of standard additions to sample analytes were better than 90%. Particular attention was paid to peak identification, including both a thorough treatment of retention time agreement of peaks in standards and sample analytes, and a comparison of results for the seven compounds amenable to quantitation by both detectors. Considerable attention was also given to determining the stability of standards and sample analytes under a wide variety of conditions, and practical recommendations were made.