Semi-micro liquid chromatography of aromatic amino acid metabolites using isocratic elution and column switching

The concentrations of catecholamine-related compounds in body fluids reflect sympathetic nerve functions. Measuring the enzyme activity of these metabolic pathways will improve diagnosis since a variety of symptoms are reported. An isocratic elution system with two column switching valves was developed using three types of semi-micro columns for fast chromatographic analysis of catecholamine related compounds. Columns are a pentyl-bonded phase, 50 x 2.1 mm i.d., a phenylhexyl-bonded phase, 100 x 2.1 mm i.d. and an octadecyl-bonded phase, 100 x 2.1 mm i.d. The separation of 20 standard compounds was achieved within 25 min using reversed-phase ion-pair liquid chromatography with an electrochemical detector. This new system was applied for analysis of catecholamine-related compounds in pig brain, since pigs are a widely used animal model for transgenic manipulation of neural genes, and MHPG (or VMA), DOPAC, DOPA, NE, EP, DA, 5HTP and 5HIAA were quantified.