title: Matrix-assisted laser desorption/ionization high-energy collision-induced dissociation of steroids: analysis of oxysterols in rat brain.
abstract: Neutral steroids have traditionally been analyzed by gas chromatography/mass spectrometry (GC/MS) after necessary derivatization reactions. However, GC/MS is unsuitable for the analysis of many conjugated steroids and those with unsuspected functional groups. Here we describe an alternative analytical method specifically designed for the analysis of oxosteroids and those with a 3beta-hydroxy-delta5 or 5alpha-hydrogen-3beta-hydroxy structure. Steroids were derivatized with Girard P (GP) hydrazine to give GP hydrazones, which are charged species and readily analyzed by matrix-assisted laser desorption/ionization mass spectrometry. The resulting [M]+ ions were then subjected to high-energy collision-induced dissociation on a tandem time-of-flight instrument. The product ion spectra give structurally informative fragment ion patterns. The sensitivity of the analytical method is such that steroid structures can be determined from low-picogram (low-femtomole) amounts of sample. The utility of the method has been demonstrated by the analysis of oxysterols extracted from rat brain.
authors.affiliation: The School of Pharmacy, University of London, 29/39 Brunswick Square, London WC1N 1AX, UK.
authors.forename: Yuqin
authors.initials: Y
authors.lastname: Wang
authors.forename: Martin
authors.initials: M
authors.lastname: Hornshaw
authors.forename: Gunvor
authors.initials: G
authors.lastname: Alvelius
authors.forename: Karl
authors.initials: K
authors.lastname: Bodin
authors.forename: Suya
authors.initials: S
authors.lastname: Liu
authors.forename: Jan
authors.initials: J
authors.lastname: Sjövall
authors.forename: William J
authors.initials: WJ
authors.lastname: Griffiths
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism*; D002853:Chromatography, Liquid; D006835:Hydrazones / Q000737:chemistry; D007664:Ketosteroids / Q000032:analysis*; D007834:Lasers; D008027:Light; D051381:Rats; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D006835:Hydrazones; D007664:Ketosteroids