Influence of the sample preparation method on the serotonin determination in plasma and platelets

Abstract

Plasma or platelet serotonin concentration is commonly used to provide information about the serotonergic activity in various psychiatric or neurological diseases. Some difficulties have been described in the measurement of serotonin (5-HT) levels in plasma or platelets. We describe an isocratic liquid-chromatographic assay with amperometric detection for determination of 5-HT in the platelet pellet and in platelet-rich and platelet-poor plasma (PRP and PPP) in sample sizes of 100 µL of plasma. The method uses an RP18 column and an amperometric detector with a thin-layer type electrochemical flow cell, with glassy carbon electrode maintained at a potential of +0.600 V vs an Ag/AgCl reference electrode. Determinations were performed in the presence or in the absence of plasma, since the biological matrix may affect the results. Different validation parameters were analysed: selectivity, accuracy, precision, linearity and stability. Reference values for 5-HT concentration in healthy adults (n = 12) were 6.6 nmol/109 platelets, for the platelet pellet, and 5.5 nmol/109 platelets, for PRP. The 100 µL sample volume used for the preparation of PPP did not make possible the determination of 5-HT levels with accuracy and precision. Copyright © 2004 John Wiley & Sons, Ltd.