Studies on neurosteroids XX. Liquid chromatography-tandem mass spectrometric method for simultaneous determination of testosterone and 5alpha-dihydrotestosterone in rat brain and serum
Autor: | Tatsuya Higashi, Kazutake Shimada, Yasuaki Ninomiya |
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Rok vydání: | 2008 |
Předmět: |
Male
Restraint Physical Spectrometry Mass Electrospray Ionization Neuroactive steroid medicine.drug_class Tandem mass spectrometry High-performance liquid chromatography Sensitivity and Specificity Analytical Chemistry Blood serum Tandem Mass Spectrometry medicine Animals Testosterone Solid phase extraction Androstanols Rats Wistar Brain Chemistry Chromatography Chemistry Reproducibility of Results General Medicine Androgen 17-Ketosteroids Rats Dihydrotestosterone Quantitative analysis (chemistry) medicine.drug Chromatography Liquid |
Zdroj: | Journal of chromatographic science. 46(7) |
ISSN: | 0021-9665 |
Popis: | Testosterone (T) and 5alpha-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-(2)H3]-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. |
Databáze: | OpenAIRE |
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