Simultaneous high-performance liquid chromatographic determination of adenosine and dopamine in rat striatal tissue with combined ultraviolet absorbance and electrochemical detection.

A method is described for the simultaneous determination of biogenic amines, adenosine and their metabolites in rat striatal tissue using high-performance liquid chromatography with ultraviolet spectrophotometric and electrochemical detection. Peaks in the chromatograms of striatal tissue extracts were identified by retention times and by on-line analysis of peak spectra for adenosine and its metabolites, and by comparing current ratios of the dual-electrode coulometric detector for monoamines and metabolites. The assay gives a linear response over the concentration range of 0.15-0.60 micrograms/ml for biogenic amines, 0.5-2.0 micrograms/ml for serotonin, 5-20 micrograms/ml for hypoxanthine, adenosine and N-methyladenosine, and 10-40 micrograms/ml for inosine. The limit of detection for striatal homogenates was 3.5 ng/g for monoamines, 9 ng/g for serotonine, 140 ng/g for hypoxanthine, 290 ng/g for inosine and 80 ng/g for adenosine. The recovery ranged from 88.5% for vanillylmandelic acid to 110.3% for dopamine. The method was used to measure biogenic amines, adenosine and related metabolites in rat striatal tissues.