A high‐performance liquid chromatographic method for simultaneous determination of free catecholamines, their basic metabolites and serotonin in human urine was developed. The compounds were separated on a precolumn of cation‐exchange resin and a mixed‐mode (C18/cation‐exchange) column and determined by an eight‐channel electrochemical detector. By this method, free norepinephrine (NE), epinephrine (E), dopamine (DA), normetanephrine (NMN), metanephrine (MN), 3‐methoxy‐tyramine (3‐MT) and serotonin (5‐HT) were determined in 40 min with inter‐assay precision of 1.9, 3.2, 1.2, 1.6, 1.6, 5.5 and 3.2%, respectively. Detection limits were between 0.05 (E) and 0.3 (3‐MT) pmol. The mean values of analytical recovery for NE, E, DA, NMN, MN, 3‐MT, 5‐HT were 99, 104, 104, 99, 102, 109 and 99% respectively.
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