Determination of 5-hydroxytryptophan, 5-hydroxytryptamine, and 5-hydroxyindoleacetic acid in 20 rat brain nuclei using liquid chromatography with electrochemical detection

1983 ◽  
Vol 61 (5) ◽  
pp. 530-534 ◽  
Author(s):  
Thérèse Di Paolo ◽  
André Dupont ◽  
Pierre Savard ◽  
Michel Daigle
Keyword(s):  
Liquid Chromatography ◽  
Rat Brain ◽  
Electrochemical Detection ◽  
Monoamine Oxidase Inhibitor ◽  
Oxidase Inhibitor ◽  
Major Metabolite ◽  
Tissue Preparation ◽  
Brain Nuclei ◽  
Hydroxyindoleacetic Acid

High-performance liquid chromatography with electrochemical detection is utilized for the simultaneous determination of serotonin, its precursor 5-hydroxytryptophan, and its major metabolite 5-hydroxyindoleacetic acid in nervous tissue samples. Tissue preparation required only homogenization in acidic solution and centrifugation prior to application to the chromatograph. Detection limits in the low picogram range were obtained for those indoles separated. This assay was used in combination with a micropunch dissection technique of 20 discrete rat brain nuclei to measure serotonin, its precursor, and major metabolite. The specificity of the assay was checked with pharmacological experiments aimed to increase or decrease serotonin levels. Pargyline, a monoamine oxidase inhibitor, led to a marked increase in serotonin and a decrease of 5-hydroxyindoleacetic acid while p-chlorophenylalanine, by blocking the conversion of tryptophan to 5-hydroxytryptophan, selectively depleted 5-hydroxytryptophan, serotonin, and 5-hydroxyindoleacetic acid.

Simplified determination of serotonin in plasma by liquid chromatography with electrochemical detection.

1984 ◽  
Vol 30 (1) ◽  
pp. 131-135 ◽  
Author(s):  
P C Tagari ◽  
D J Boullin ◽  
C L Davies
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Human Plasma ◽  
Electrochemical Detection ◽  
High Performance ◽  
Carcinoid Syndrome ◽  
Quantitative Detection ◽  
Chromatographic Purification ◽  
Hydroxyindoleacetic Acid

Abstract This simplified determination of serotonin in human plasma involves "high-performance" liquid chromatography with electrochemical detection. Deproteinated samples of platelet-poor plasma are injected directly into the chromatograph without preliminary chromatographic purification. The limit of quantitative detection of serotonin in plasma is 5 nmol/L (equivalent to 250 fmol injected). Intra- and inter-assay CVs ranged from 5.6% to 7.7% and 3.5% to 14.9%, respectively, for concentrations of serotonin from 0.02 to 10.0 mumol/L. Results agreed well with those by spectrofluorometry, both for plasma standards (r = 0.985, range 100 nmol/L to 100 mumol/L) and for pathological samples obtained from four carcinoid syndrome patients (r = 0.964). The method is also applicable to the concurrent measurement of 5-hydroxytryptophan and 5-hydroxyindoleacetic acid.

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