title: Determination of seven selected neuro- and immunomodulatory steroids in human cerebrospinal fluid and plasma using LC-MS/MS.
abstract: Dehydroepiandrosterone (DHEA) and its 7-oxo- and 7-hydroxy-metabolites occurring in the brain are considered neurosteroids. Metabolism of the latter is catalysed by 11β-hydroxysteroid dehydrogenase (11β-HSD) which also interconverts cortisol and cortisone. The concurrent metabolic reaction to DHEA 7-hydroxylation is the formation of 16α-hydroxy-DHEA. The LC-MS/MS method using triple stage quadrupole-mass spectrometer was developed for simultaneous quantification of free DHEA, 7α-hydroxy-DHEA, 7β-hydroxy-DHEA, 7-oxo-DHEA, 16α-hydroxy-DHEA, cortisol and cortisone in human plasma and cerebrospinal fluid (CSF). The method employs 500 μL of human plasma and 3000 μL of CSF extracted with diethyl ether and derivatized with 2-hydrazinopyridine. It has been validated in terms of sensitivity, precision and recovery. In plasma, the following values were obtained: limit of detection: 2-50p g/mL; limit of quantification: 5-140 pg/mL; within-day precision 0.58-14.58%; between-day precision: 1.24-13.89% and recovery: 85-113.2%). For CSF, the values of limit of detection: 2-28 pg/mL; limit of quantification: 6-94 pg/mL; within-day precision; 0.63-5.48%; between-day precision: 0.88-14.59% and recovery: 85.1-109.4% were acquired. Medians and concentration ranges of detected steroids in plasma and CSF are given in subjects with excluded normal pressure hydrocephalus (n=37; 65-80 years). The method enables simultaneous quantification of steroids important for the estimation of 11β-HSD activity in human plasma and CSF. It will be helpful in better understanding various degenerative diseases development and progression.
authors: Institute of Endocrinology, Department of Steroids and Proteofactors, Narodni 8, 11694 Prague, Czech Republic.
authors: Lucie
authors: L
authors: Sosvorova
authors: Institute of Endocrinology, Department of Steroids and Proteofactors, Narodni 8, 11694 Prague, Czech Republic.
authors: Jana
authors: J
authors: Vitku
authors: Institute of Endocrinology, Department of Steroids and Proteofactors, Narodni 8, 11694 Prague, Czech Republic.
authors: Tereza
authors: T
authors: Chlupacova
authors: Military University Hospital Prague, Department of Neurosurgery, U Vojenske nemocnice 1200, 16902 Prague, Czech Republic.
authors: Milan
authors: M
authors: Mohapl
authors: Institute of Endocrinology, Department of Steroids and Proteofactors, Narodni 8, 11694 Prague, Czech Republic. Electronic address: rhampl@endo.cz.
authors: Richard
authors: R
authors: Hampl
mesh_terms: D042842:11-beta-Hydroxysteroid Dehydrogenases / Q000378:metabolism; D000368:Aged; D000369:Aged, 80 and over; D003687:Dehydroepiandrosterone / Q000031:analogs & derivatives* / Q000134:cerebrospinal fluid*; D005260:Female; D006801:Humans; D007155:Immunologic Factors / Q000134:cerebrospinal fluid*; D008297:Male; D018377:Neurotransmitter Agents / Q000134:cerebrospinal fluid*
chemical_list: D007155:Immunologic Factors; D018377:Neurotransmitter Agents; D003687:Dehydroepiandrosterone; D042842:11-beta-Hydroxysteroid Dehydrogenases
keywords: 11β-Hydroxysteroid dehydrogenase; Cerebrospinal fluid; Dehydroepiandrosterone metabolites; LC–MS/MS