title: Precise and accurate assay of pregnenolone and five other neurosteroids in monkey brain tissue by LC-MS/MS.
abstract: A series of steroids present in the brain have been named "neurosteroids" following the possibility of their role in the central nervous system impairments such as anxiety disorders, depression, premenstrual dysphoric disorder (PMDD), addiction, or even neurodegenerative disorders such as Alzheimer's and Parkinson's diseases. Study of their potential role requires a sensitive and accurate assay of their concentration in the monkey brain, the closest model to the human. We have thus developed a robust, precise and accurate liquid chromatography-tandem mass spectrometry method for the assay of pregnenolone, pregnanolone, epipregnanolone, allopregnanolone, epiallopregnanolone, and androsterone in the cynomolgus monkey brain. The extraction method includes a thorough sample cleanup using protein precipitation and phospholipid removal, followed by hexane liquid-liquid extraction and a Girard T ketone-specific derivatization. This method opens the possibility of investigating the potential implication of these six steroids in the most suitable animal model for neurosteroid-related research.
authors.affiliation: Endoceutics Inc., 2795 Laurier Blvd, Suite 500, Quebec City (QC) G1V 4M7, Canada.
authors.forename: Alain Y
authors.initials: AY
authors.lastname: Dury
authors.affiliation: Endoceutics Inc., 2795 Laurier Blvd, Suite 500, Quebec City (QC) G1V 4M7, Canada.
authors.forename: Yuyong
authors.initials: Y
authors.lastname: Ke
authors.affiliation: Endoceutics Inc., 2795 Laurier Blvd, Suite 500, Quebec City (QC) G1V 4M7, Canada. Electronic address: fernand.labrie@endoceutics.com.
authors.forename: Fernand
authors.initials: F
authors.lastname: Labrie
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism*; D002853:Chromatography, Liquid; D000882:Haplorhini; D007659:Ketones / Q000032:analysis; D015394:Molecular Structure; D018377:Neurotransmitter Agents / Q000032:analysis*; D011284:Pregnenolone / Q000032:analysis*; D053719:Tandem Mass Spectrometry
chemical_list: D007659:Ketones; D018377:Neurotransmitter Agents; D011284:Pregnenolone
keywords: Cynomolgus monkey; Girard T reagent; Liquid chromatography–tandem mass spectrometry; Neurosteroids