Simultaneous determination of levodopa methyl ester, levodopa, 3-O-methyldopa and dopamine in plasma by high-performance liquid chromatography with electrochemical detection.

A new procedure is described for the simultaneous determination of levodopa methyl ester (LDME) and its biotransformation products levodopa (L-DOPA), 3-O-methyldopa (3-OMD) and dopamine (DA) in stabilized plasma samples, using reversed-phase high-performance liquid chromatography. A coulometric detector equipped with a dual-electrode system operating in the redox mode was used to simultaneously quantitate all compounds. This system generated a double signal monitored by a dual-channel acquisition data system and allowed quantitation of compounds at the nanogram level. The intra- and inter-assay precision varied in the 2.4-6.9% and 3.2-9.1% ranges respectively, whereas the recoveries were close to 85% for L-DOPA and 3-OMD and 70% for DA and LDME. Samples may be stored at -80 degrees C for 15 days before analysis. The method was applied to plasma samples after oral administration of LDME to rats, but it may also be suitable for human pharmacokinetic studies.