Quantitation of Caffeine by Capillary Zone Electrophoresis with End-Column Amperometric Detection at a Carbon Microdisk Array Electrode

Published: Journal of Chromatographic Science, ISSN 0021-9665 Volume 38, Number 1, January 2000, pp. 6-10

Studies on Neurosteroids X. Determination of Pregnenolone and Dehydroepiandrosterone in Rat Brains Using Gas Chromatography–Mass Spectrometry–Mass Spectrometry
Kazutake Shimada and Kenichiro Yago
Faculty of Pharmaceutical Sciences, Kanazawa University, 13-1 Takara-machi, Kanazawa 920-0934, Japan

An assay method for pregnenolone and dehydroepiandrosterone in rat brains is developed using gas chromatography (GC)–electron ionization-mass spectrometry (MS)–MS. The extract of the rat brain homogenate containing deuterated internal standard with organic solvent is purified by silica gel minicolumn chromatography. The obtained fraction is derivatized into methyloxime, treated with dimethylisopropylsilylimidazole, and then subjected to GC–MS–MS. The method is applied to the determination of these steroids in the gray matter and olfactory bulbs of rat brains, which are divided into control and acute stressed specimens. Although pregnenolone in both regions of the rat brains increases more than three times after stress, dehydroepiandrosterone in both regions is not so clearly influenced by stress.

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