Analytical considerations for quantitative determination of serotonin and its metabolically related products in biological matrices.
We discuss the use of "high-performance" liquid chromatography with electrochemical detection for the quantification fo serotonin (5-hydroxytryptamine) and metabolically related products (i. e., 5-hydroxyindoleacetic acid and 5-hydroxytryptophan) in biological matrices. Two methods are described: one for routine quantification of nanogram amounts or more; the other a two-column purification-separation ("trace enrichment") technique for determining subnanogram amounts of these compounds. Various factors affecting the liquid-chromatographic separation and quantification of 5-hydroxy-indoles are detailed, including selection of appropriate internal standard, sample purification, and effects of changes in mobile phase composition, e.g., ion-pair reagent, organic modifier content, and pH. We describe in detail the trace-enrichment method for determinations in the range of 25 pg to 25 ng, and discuss the general factors to consider when developing or modifying an assay for specific applications.
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D006916:5-Hydroxytryptophan / Q000032:analysis*; D001923:Brain Chemistry; D002851:Chromatography, High Pressure Liquid; D004563:Electrochemistry; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006897:Hydroxyindoleacetic Acid / Q000032:analysis*; D008168:Lung / Q000032:analysis; D008832:Microchemistry; D012701:Serotonin / Q000032:analysis*; D013048:Specimen Handling; D014599:Uterus / Q000032:analysis
D012701:Serotonin; D006897:Hydroxyindoleacetic Acid; D006916:5-Hydroxytryptophan