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
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), Vol. 7:1
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
HYDROXYTRYPTOPHAN
BIOCHEMISTRY
SEROTONIN
TRITIUM COMPOUNDS
BRAIN
ISOTOPE APPLICATIONS
LIQUID COLUMN CHROMATOGRAPHY
RADIOCHROMATOGRAPHY
RATS
AMINES
AMINO ACIDS
ANIMALS
AUTONOMIC NERVOUS SYSTEM AGENTS
AZOLES
BODY
CARBOXYLIC ACIDS
CENTRAL NERVOUS SYSTEM
CHEMISTRY
CHROMATOGRAPHY
DRUGS
HETEROCYCLIC COMPOUNDS
HYDROXY ACIDS
HYDROXY COMPOUNDS
INDOLES
LABELLED COMPOUNDS
MAMMALS
NERVOUS SYSTEM
NEUROREGULATORS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANS
PYRROLES
RADIOPROTECTIVE SUBSTANCES
RODENTS
SEPARATION PROCESSES
SYMPATHOMIMETICS
TRYPTAMINES
VERTEBRATES
550201* - Biochemistry- Tracer Techniques
551001 - Physiological Systems- Tracer Techniques