Determination of serotonin, melatonin and metabolites in gastrointestinal tissue using high-performance liquid chromatography with electrochemical detection

In this paper we show a simple isocratic chromatographic method for the detection of serotonin and its precursors and metabolites from various types of gastrointestinal tissue. The paper measures for the first time basal measurements of melatonin in the gastrointestinal tract, which has recently been shown to be released from the musosal lining of the gut. Tissue samples were stable following sample preparation in either 0.1 m perchloric acid or mobile phase. Analysis was carried out using a mobile phase consisting of 10% acetonitrile-90% acetate acid buffer pH 4.0 with 2 mm decane-sulfonic acid sodium salt at a column temperature of 50 degrees C. Electrochemical detection was utilized at a potential of +850 mV vs Ag/AgCl reference electrode at 10 microA full-scale deflection. The detection limit of 5-HT and melatonin was 241 and 308 nm respectively for a 10 microL injection. As a result of the method optimization, total analysis was reduced to 30 min. Accurate responses of the tissue samples following sample preparation could be obtained following a week after storage at -80 degrees C. This method is capable of preparing and analysing of samples from all regions of the gastrointestinal tract.