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 language | English |
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Pages (from-to) | 269-276 |
Number of pages | 8 |
Journal | Journal of Chromatography B |
Volume | 818 |
Issue number | 2 |
DOIs | |
Publication status | Published - Apr 2005 |
Keywords
- Microbore HPLC
- Serotonin
- Catecholamines
- Lymnaea stagnalis