Liquid Chromatographic determination of homovanillic acid, 5-hydroxyindoleacetic acid and probenecid levels in human cerebrospinal fluid during probenecid test

Abstract

Homovanillic acid and 5-hydroxyindoleacetic acid, respectively the major metabolites of the central neurotransmitters dopamine and serotonin, are present in human lumbar cerebrospinal fluid, and their determination during the probenecid test is used to study brain monoamine abnormalities in man. We developed a high performance liquid Chromatographie technique coupled with electrochemical detection which allows the simultaneous determination of homovanillic acid and 5-hydroxyindoleacetic acid on 10 μl of human lumbar cerebrospinal fluid. Another liquid Chromatographie technique coupled with ultra-violet detection was applied to the measurement of probenecid after injection of 10 μl of native cerebrospinal fluid. The main advantages of these two techniques are their simplicity, sensitivity, rapidity (five cerebrospinal fluid samples are analysed within one hour) and their good day-to-day reproducibility. These methods were applied to probenecid tests performed in several neurological patients.