Serotonin (5-hydroxytryptamine, 5-HT) is a monoamine neurotransmitter synthesized in serotonergic neurons in the Central Nervous System (CNS) and in enterochromaffin cells in the gastrointestinal tract. One product of Serotonin breakdown is 5-hydroxyindoleacetic acid (5-HIAA) which is excreted in the urine. Low levels of Serotonin may be associated with several disorders, namely clinical depression, obsessive-compulsive disorder and anxiety disorders. There are several abuse drugs (i.e. Cocaine, LSD, Amphetamines) that modulate the 5-HT system and can lead to addiction-related behavior in humans. The increase in 5-HT levels is one of the core mechanisms of action for drug addiction, although the dopaminergic system still plays an important role in the investigation of abuse drug behavioral effects. The monitoring of monoamines and some metabolites in human plasma is a useful clinical research tool for examining the etiologies of neuroendocrine disorders and the involvement of dopaminergic and serotoninergic activity under different physiological or pathological conditions (i.e. addiction phenomena). An HPLC method with coulometric detection is currently under development for the assay of Serotonin, 5-HIAA and Dopamine metabolites (3,4-dihydroxyphenylacetic acid or DOPAC, and 4-hydroxy-3-methoxyphenylacetic acid or HVA). All compounds are separated on an RP C18 column with a mobile phase composed of MeOH and a pH 3.0 citrate buffer also containing octanesulfonic acid and EDTA. Electrochemical detection is carried out using a coulometric detector in oxidation mode. An original and fast procedure of plasma pretreatment based on deproteinization procedure with perchloric acid has been implemented and has provided high extraction yields (>90%) for all analytes. The method seems to be suitable for the determination of Serotonin, 5 HIAA, DOPAC and HVA levels in human plasma from former drug users.
M.A. Saracino, M. Vitali, A. Ferranti, G. Gerra, C. Leonardi, M.A. Raggi (2007). Simultaneous determination of monoamine neurotransmitters and metabolites by high performance liquid chromatography with coulometric detection. FIRENZE : RDPA.
Simultaneous determination of monoamine neurotransmitters and metabolites by high performance liquid chromatography with coulometric detection
SARACINO, MARIA ADDOLORATA;FERRANTI, ANNA;RAGGI, MARIA AUGUSTA
2007
Abstract
Serotonin (5-hydroxytryptamine, 5-HT) is a monoamine neurotransmitter synthesized in serotonergic neurons in the Central Nervous System (CNS) and in enterochromaffin cells in the gastrointestinal tract. One product of Serotonin breakdown is 5-hydroxyindoleacetic acid (5-HIAA) which is excreted in the urine. Low levels of Serotonin may be associated with several disorders, namely clinical depression, obsessive-compulsive disorder and anxiety disorders. There are several abuse drugs (i.e. Cocaine, LSD, Amphetamines) that modulate the 5-HT system and can lead to addiction-related behavior in humans. The increase in 5-HT levels is one of the core mechanisms of action for drug addiction, although the dopaminergic system still plays an important role in the investigation of abuse drug behavioral effects. The monitoring of monoamines and some metabolites in human plasma is a useful clinical research tool for examining the etiologies of neuroendocrine disorders and the involvement of dopaminergic and serotoninergic activity under different physiological or pathological conditions (i.e. addiction phenomena). An HPLC method with coulometric detection is currently under development for the assay of Serotonin, 5-HIAA and Dopamine metabolites (3,4-dihydroxyphenylacetic acid or DOPAC, and 4-hydroxy-3-methoxyphenylacetic acid or HVA). All compounds are separated on an RP C18 column with a mobile phase composed of MeOH and a pH 3.0 citrate buffer also containing octanesulfonic acid and EDTA. Electrochemical detection is carried out using a coulometric detector in oxidation mode. An original and fast procedure of plasma pretreatment based on deproteinization procedure with perchloric acid has been implemented and has provided high extraction yields (>90%) for all analytes. The method seems to be suitable for the determination of Serotonin, 5 HIAA, DOPAC and HVA levels in human plasma from former drug users.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.