title: Synthesis of 3α-deuterated 7α-hydroxy-DHEA and 7-oxo-DHEA and application in LC-MS/MS plasma analysis.
abstract: 7-Oxygenated metabolites of dehydroepiandrosterone (DHEA) are known for their neuroprotective and immunomodulatory properties. These neuroactive steroids are currently predominately analysed by mass spectrometry, for which the use of internal deuterated standards is necessary. The aim of this study was to synthesize the deuterated derivatives of 7α-hydroxy-DHEA and 7-oxo-DHEA and test them in liquid chromatography-tandem mass spectrometry (LC-MS/MS) in order to enhance the performance characteristics of this method. Here we report the synthesis of 3α deuterium-labelled 7α-hydroxy-DHEA and 7-oxo-DHEA. Deuterium was introduced into the 3α position by reduction of the corresponding 3-ketone with a protected 17-carbonyl group using NaBD4. Our new procedure allows the easier synthesis of deuterated steroid labelled compounds. The use of these deuterated steroids enabled us to improve the human plasma LC-MS/MS analysis of 7α-hydroxy-DHEA and 7-oxo-DHEA in terms of sensitivity, precision and recovery.
authors.affiliation: Institute of Endocrinology, Department of Steroids and Proteofactors, Narodni 8, 116 94 Prague, Czech Republic. Electronic address: lsosvorova@endo.cz.
authors.forename: Lucie
authors.initials: L
authors.lastname: Kolatorova Sosvorova
authors.affiliation: Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacky University, Hnevotinska 5, 77900 Olomouc, Czech Republic. Electronic address: jan.sarek@gmail.com.
authors.forename: Jan
authors.initials: J
authors.lastname: Sarek
authors.affiliation: Institute of Endocrinology, Department of Steroids and Proteofactors, Narodni 8, 116 94 Prague, Czech Republic. Electronic address: jvitku@endo.cz.
authors.forename: Jana
authors.initials: J
authors.lastname: Vitku
authors.affiliation: Laboratory of Growth Regulators, Centre of the Region Hana for Biotechnological and Agricultural Research, Institute of Experimental Botany ASCR & Palacky University, Slechtitelu 27, 78371 Olomouc, Czech Republic. Electronic address: miroslav.kvasnica@upol.cz.
authors.forename: Miroslav
authors.initials: M
authors.lastname: Kvasnica
mesh_terms: D002853:Chromatography, Liquid; D003687:Dehydroepiandrosterone / Q000031:analogs & derivatives* / Q000138:chemical synthesis* / Q000737:chemistry; D003903:Deuterium / Q000737:chemistry*; D015394:Molecular Structure; D053719:Tandem Mass Spectrometry
chemical_list: C002914:7-oxodehydroepiandrosterone; D003687:Dehydroepiandrosterone; D003903:Deuterium
keywords: 7-Oxo-DHEA; 7α-Hydroxy-DHEA; Deuterated standard; Mass spectrometry; Neurosteroid; Steroid