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Several neuropsychopharmacological properties have been attributed to the 3α-reduced pregnane steroids, allopregnanolone and pregnanolone, as well as to dehydroepiandrosterone sulfate because of their ability to modulate γ-aminobutyric acid (GABA A ) receptors in the CNS. In order to understand better their role in several mechanisms in CNS, a new methodology is proposed to monitor these compounds in human plasma. Methodology &amp; results: The analytes were first derivatized with 2-hydrazinopyridine and extracted from plasma using SPE. Then, the compounds were separated and detected by LC-MS/MS. A mobile phase of formic acid (0.1%) in water and methanol through a gradient of composition and a flow rate of 0.3 ml min -1 resulted in good separations of the analytes. Linear responses in wide range of concentrations and LOQs ranging from 10 (dehydroepiandrosterone 3-sulfate) to 40 pg ml -1 (dehydroepiandrosterone) were obtained in &lt;9 min. The method proposed has been validated and then applied to monitor these neurosteroids in plasma samples from ten volunteers.

bioanalysis

LC–MS/MS Simultaneous Analysis of Allopregnanolone, Epiallopregnanolone, Pregnanolone, Dehydroepiandrosterone and Dehydroepiandrosterone 3-Sulfate in Human Plasma