Serotonin is a neurotransmitter and vasoconstrictor that has been implicated in a variety of conditions and physiological
responses such as essential hypertension, vascular pain and emesis. The effective analysis of whole blood serotonins, therefore,
is necessary to monitor the role serotonin regulation has upon these and other sympathoadrenal-mediated events.
Here in our research lab, HPLC with electrochemical detection is used to determine whole blood serotonin levels. Serotonin,
5-hydroxytryptamine (5-HT), is easily distinguishable from the internal standard, N-w-methyl-5-hydroxytryptamine (5-MHT).
Both monoamines exhibit good chromatographic quality under our current run conditions.
During sample preparation, the addition of carbon monoxide is critical to the preservation of serotonins upon lysis. Whole
blood serotonin values (ng/ml) are calculated for individual samples by means of a linear regression analysis, based upon
a triplicate injection of a single-point external reference standard (free base: 40ng/ml 5-HT; 40ng/ml 5-MHT). In addition
5-MHT is used as an internal standard. The limit of detection for our assay is 1.875ng/ml.
