A rapid and simple method to determine the specific activities of serotonin, 5-hydroxyindoleacetic acid, and 5-hydroxytryptophan in brain by HPLC with electrochemical detection

Journal Article · Fri Jan 01 04:00:00 EST 1982 · Neurochem. Res.; (United States)

DOI:https://doi.org/10.1007/BF00965071· OSTI ID:7089879

Bernstein, M J; Shea, P A

The specific activities of 5-hydroxytryptophan (5-HTP), serotonin (5-HT), and 5-hydroxyindoleacetic acid (5-HIAA) have been determined in the brain of rats by HPLC using electrochemical detection. The method allows, from a single sample, the simultaneous measurement of all three compounds and collection of each peak for radioactivity determinations. Five male Wistar rats were injected i.v. with 2.0 mCi/kg of DL-5-hydroxy-(G-/sup 3/H)tryptophan (2.6 Ci/mmol) and 30 min later the animals were killed by near freezing. Whole brains were removed and homogenized in an acid medium. The content of 5-HTP, 5-HT, and 5-HIAA were determined by HPLC. Each peak of interest was immediately collected after detection in scintillation vials by use of a small dead space detector (TL-9A, Bioanalytical Systems, Inc.). The amounts of radioactivity were determined and specific activities calculated from the results. A second chromatography system (TLC) was used to check the authenticity and purity of compounds separated by the HPLC.

• Research Organization: Indiana University School of Medicine, Indianapolis
• OSTI ID: 7089879
• Journal Information: Neurochem. Res.; (United States), Journal Name: Neurochem. Res.; (United States) Vol. 7:1; ISSN NERED
• Country of Publication: United States
• Language: English

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59 BASIC BIOLOGICAL SCIENCES
AMINES
AMINO ACIDS
ANIMALS
AUTONOMIC NERVOUS SYSTEM AGENTS
AZOLES
BIOCHEMISTRY
BODY
BRAIN
CARBOXYLIC ACIDS
CENTRAL NERVOUS SYSTEM
CHEMISTRY
CHROMATOGRAPHY
DRUGS
HETEROCYCLIC COMPOUNDS
HYDROXY ACIDS
HYDROXY COMPOUNDS
HYDROXYTRYPTOPHAN
INDOLES
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LIQUID COLUMN CHROMATOGRAPHY
MAMMALS
NERVOUS SYSTEM
NEUROREGULATORS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANS
PYRROLES
RADIOCHROMATOGRAPHY
RADIOPROTECTIVE SUBSTANCES
RATS
RODENTS
SEPARATION PROCESSES
SEROTONIN
SYMPATHOMIMETICS
TRITIUM COMPOUNDS
TRYPTAMINES
VERTEBRATES