Journal Article

Studies on Neurosteroids XX. Liquid Chromatography—Tandem Mass Spectrometric Method for Simultaneous Determination of Testosterone and 5α-Dihydrotestosterone in Rat Brain and Serum

Tatsuya Higashi, Yasuaki Ninomiya and Kazutake Shimada

in Journal of Chromatographic Science

Volume 46, issue 7, pages 653-658
Published in print August 2008 | ISSN: 0021-9665
Published online August 2008 | e-ISSN: 1945-239X | DOI: http://dx.doi.org/10.1093/chromsci/46.7.653
Studies on Neurosteroids XX. Liquid Chromatography—Tandem Mass Spectrometric Method for Simultaneous Determination of Testosterone and 5α-Dihydrotestosterone in Rat Brain and Serum

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Testosterone (T) and 5α-dihydrotestosterone (DHT) are now referred to not only as androgenic steroid hormones, but also as neuroactive steroids, because they elicit anesthetic and anxiolytic effects. Methods using liquid chromatography—electrospray ionization-tandem mass spectrometry (LC—ESI-MS—MS) coupled with derivatization are developed and validated to examine rat brain and serum levels of T and DHT and their stress-induced changes. The steroids are extracted with methanol—acetic acid from the brain tissue or serum, purified using solid-phase extraction cartridges, derivatized with a permanently charged reagent, 2- hydrazino-1-methylpyridine, and subjected to LC—MS—MS. [19,19,19-2H3]-T is used as the internal standard. The intra- and inter-assay coefficients of variation are below 10.0%, and the analytical recoveries are 98.1–103.0%. The developed methods are applied to the animal study and it was found that a fair amount of DHT is continuously and locally synthesized in the brain, and its level is not changed by the immobilization stress and depends on the brain T level.

Journal Article.  0 words. 

Subjects: Chemistry

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