The clinical use of positron emission tomography, PET, with selected radiolabelled tracer molecules visualizing and quantitating physiological processes in the tissue relies in many situations on compartmental models for the interpretation of the radiosignal. Validation of such models must, therefore, include chromatographic analysis of the radioactivity composition of the signal. Rapid and sensitive liquid chromatographic methods amenable for automation for the analysis of [11C] labelled L-DOPA and its metabolites were therefore developed and validated for the quantitation of radioactivity composition in rat brain microdialysates as well as homogenates. Analysis included a simple isolation step, separation using reversed phase liquid chromatography with radiometric detection and permitted assay following tracer doses with an analysis time of 15 min. The analysis of radioactivity composition in the rat striatum showed that peripherally formed O-methyl L-DOPA constituted less than 20% of the radioactivity 40 min after injection of L-[beta-11C]DOPA. In the extracellular space the main component was [11C]-homovanillic acid which increased with time indicating rapid formation but slow elimination. The cumulation of radioactivity in the striatum corresponded to the radioactivity signal of dopamine and derived metabolites. The formation rate of dopamine in the rat corresponded closely to the utilization rate in the striatum of monkey and man measured with PET. This indicated that the rate constants measured with PET correlates well to the dopamine synthesis rate.