High-performance liquid chromatographic determination of urinary free catecholamines with electrochemical detection...

A reliable high-performance liquid chromatographic method is presented for the determination of the urinary free catecholamines noradrenaline, adrenaline and dopamine. Urine is purified on a column of immobilized boric acid. Catecholamines are separated by ion-pair reversed phase high-performance liquid chromatography and detected electrochemically. The method is suited for routine analysis. It allows the determination of urinary free catecholamines in concentrations as low as 1 microgram/1 for noradrenaline and adrenaline and 5 micrograms/1 for dopamine. A single analysis can be completed within 1 h. Routine analyses can be carried out in a series of 40 samples within 2 days. The within-assay and between-assay coefficients of variation of the analyses in urine were both 2.9% for noradrenaline, both 5.0% for adrenaline, and 1.9 and 2.1% for dopamine. The chromatographic properties of the immobilized boric acid were investigated. In particular, the elution pattern of a series of catecholamine metabolites and analogues was determined. Under the conditions used, only basic compounds containing both a vicinal hydroxyl configuration and a primary or secondary amino group adsorb and elute together with the free catecholamines.