Reversed-phase liquid chromatography with amperometric detection of lipophilic dopamine analogues and determination of brain and serum concentrations after sample clean-up on small sephadex G-10 columns

The liquid chromatographic determination of N-alkylated analogues of dopamine is described. The retention and separation of these compounds, ranging from dopamine to N,N-dibutyl-dopamine, was studied on four bonded-phase columns, of which Nucleosil 5 C₁, was chosen for routine use. The compounds were detected by a rotating disc amperometric detector. Samples of rat brain and serum were taken through a clean-up step on small Sephadex G-10 columns from which the dopamine analogues eluted in the same fraction as dopamine. The overall recovery was 70–90% from brain tissue and 60–70% from serum or plasma. The limit of detection for the catechol-containing compounds in tissue was 40–100 pg, for O-methylated ones 100–200 pg. The method is applied to the determination of dopamine analogues in rat brain after peripheral administration.