Simultaneous determination of four monoamine metabolites and serotonin in cerebrospinal fluid by 'high-performance' liquid chromatography with electrochemical detection; application for patients with Alzheimer's disease

Abstract

We describe an optimized, sensitive assay procedure for simultaneously determining 3-methoxy-4-hydroxyphenylglycol, 3,4-dihydroxyphenylacetic acid, homovanillic acid, 5-hydroxyindoleacetic acid, and serotonin in human lumbar cerebrospinal fluid. The assay is based on liquid chromatography with an electrochemical detection system equipped with a newly developed, highly sensitive graphite electrode. Before assay, samples are deproteinized with perchloric acid, 0.2 mol/L. The within- and between-day CVs were less than 2.5% nd 6.7%, respectively, for all compounds. The lower limits of detection for 3-methoxy-4-hydroxyphenylglycol and 3,4-dihydroxyphenylacetic acid were 0.5 pg and for the other three analytes 1.5 pg per injection. Preliminary data on monoaminergic metabolites by the current assay showed that concentrations of dopaminergic and serotonergic metabolites were significantly (P < 0.05) lower in patients with Alzheimer's disease than in the age-matched controls. In addition, the concentrations of 3,4-dihydroxyphenylacetic acid correlated significantly (P < 0.01) with that of homovanillic acid, suggesting that monitoring either of these metabolites in cerebrospinal fluid can be used as an index of central dopaminergic activity.