Simple automated high-performance liquid chromatographic column-switching technique for the measurement of dopa and 3-O-methyldopa in plasma.

A reliable assay procedure for the determination of 3,4-dihydroxyphenyl-L-alanine (dopa) and its O-methylated metabolite 3-O-methyldopa (3-OMD) is described. Supernatants from deproteinized plasma samples were directly injected into a column-switching system, allowing on-line prepurification of the samples by cation-exchange chromatography followed by reversed-phase high-performance liquid chromatography with electrochemical detection. With this method, the detector response was linear from endogenous levels of dopa and 3-OMD up to microgram concentrations. This technique combines the simplicity of direct injection methods with advantages of procedures using a sample clean-up. It represents a valid tool for the rapid and accurate measurement of large numbers of plasma samples in animals treated with levodopa and in clinical trials with new levodopa formulations.