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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

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Abstract

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 ofDl-5-hydroxy-[G-3H]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.

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Authors and Affiliations

  1. Indiana University School of Medicine, 46223, Indianapolis, Indiana

    M. J. Bernstein

  2. Institute of Psychiatric Research Departments of Psychiatry and Biochemistry, Indiana University School of Medicine, Indianapolis, Indiana

    P. A. Shea

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  1. M. J. Bernstein
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  2. P. A. Shea
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Bernstein, M.J., Shea, P.A. 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. Neurochem Res 7, 79–85 (1982). https://doi.org/10.1007/BF00965071

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  • DOI: https://doi.org/10.1007/BF00965071

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