Simple and rapid determination of serotonin and catecholamines in biological tissue using high-performance liquid chromatography with electrochemical detection

Bhavik Patel, M. Arundell, K.H. Parker, Mark Yeoman, D. O'Hare

Research output: Contribution to journalArticle

Abstract

Using the CNS of Lymnaea stagnalis a method is described for the rapid analysis of neurotransmitters and their metabolites using high performance liquid chromatography coupled with electrochemical detection. Tissue samples were homogenised in ice-cold 0.1 M perchloric acid and centrifuged. Using a C18 microbore column the mobile phase was maintained at a flow rate of 100 μl/min and consisted of sodium citrate buffer (pH 3.2)–acetonitrile (82.5:17.5, v/v) with 2 mM decane-sulfonic acid sodium salt. The potential was set at +750 mV versus Ag|AgCl reference electrode at a sensitivity of 50 nA full scale deflection. The detection limit for serotonin was 11.86 ng ml−1 for a 5 μl injection. Preparation of tissue samples in mobile phase reduced the response to dopamine and serotonin compared with perchloric acid. In addition it was found that the storage of tissue samples at −20 °C caused losses of dopamine and serotonin. As a result of optimising the sample preparation and mobile phase the total time of analysis was substantially reduced resulting in a sample preparation and assay time of 15–20 min.
Original languageEnglish
Pages (from-to)269-276
Number of pages8
JournalJournal of Chromatography B
Volume818
Issue number2
DOIs
Publication statusPublished - Apr 2005

Keywords

  • Microbore HPLC
  • Serotonin
  • Catecholamines
  • Lymnaea stagnalis

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