Determination of free and total catecholamines and salsolinol in urine by ion-pair reversed-phase liquid chromatography with electrochemical detection after a one-step sample clean-up.

A simple method is presented for the routine determination of free and total catecholamines and salsolinol in urine. For the analysis of total catecholamines and salsolinol, the urine samples are first hydrolysed. The compounds are isolated by a one-step sample clean-up by cation-exchange chromatography and separated by ion-pair reversed-phase liquid chromatography and detected electrochemically. The method is suited for routine analysis. Pretreatment of ten samples for the high-performance liquid chromatographic analysis of free and total catecholamines and salsolinol is carried out in 4 h. A single analysis is completed in 30 min. The absolute losses of the compounds were less than 10%, except for epinephrine (20%). The detection limits were 0.9, 2.4, 4.8 and 6 nmol/l for norepinephrine, epinephrine, dopamine and salsolinol, respectively, in native urine. The values were twice as high for hydrolysed urine. The sample clean-up appears to be rather specific: apart from catecholamines and salsolinol, only those structurally related amines that contain a vicinal hydroxyl group are isolated.