title: LC-MS/MS simultaneous analysis of allopregnanolone, epiallopregnanolone, pregnanolone, dehydroepiandrosterone and dehydroepiandrosterone 3-sulfate in human plasma.
abstract: OBJECTIVE
  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 (GABAA) 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 & 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-1resulted 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 <9 min. The method proposed has been validated and then applied to monitor these neurosteroids in plasma samples from ten volunteers.
  
  
  CONCLUSIONS
  For the first time, a straightforward and reliable method for the chromatographic separation of allopregnanolone, epiallopregnanolone and pregnanolone, as well as of dehydroepiandrosterone and dehydroepiandrosterone 3-sulfate was carried out, with optimal accuracy, sensitivity and specificity.
authors.affiliation: Advanced Molecular Diagnostics Unit, Istituto Dermopatico dell'Immacolata (IDI-IRCCS), Rome 00167, Italy.
authors.forename: Luana
authors.initials: L
authors.lastname: Lionetto
authors.affiliation: CICAB Clinical Research Centre, Extremadura University Hospital & Medical School, Badajoz 06071, Spain.|Department of Analytical Chemistry & Food Technology, Faculty of Pharmacy, University of Castilla-La Mancha, Albacete 02071, Spain.
authors.forename: Fernando
authors.initials: F
authors.lastname: De Andrés
authors.affiliation: Laboratory of Experimental Immunology, Istituto Dermopatico dell'Immacolata (IDI-IRCCS), Rome 00167, Italy.
authors.forename: Matilde
authors.initials: M
authors.lastname: Capi
authors.affiliation: Psychiatry & Neurology Department, Sapienza University of Rome, Rome 00185, Italy.|Department of Psychiatry, Harvard Medical School, Boston, MA 02215, USA.|Department of Molecular Medicine, Sant'Andrea Medical Center, Italy Regional Referral Headache Center, Sant'Andrea Hospital, Sapienza University of Rome, Rome 00189, Italy.
authors.forename: Martina
authors.initials: M
authors.lastname: Curto
authors.affiliation: NESMOS Department, Sapienza University of Rome, Rome00189, Italy.
authors.forename: Donata
authors.initials: D
authors.lastname: Sabato
authors.affiliation: NESMOS Department, Sapienza University of Rome, Rome00189, Italy.
authors.forename: Maurizio
authors.initials: M
authors.lastname: Simmaco
authors.affiliation: Experimental Neuro-Psychobiology Lab, Department of Clinical & Behavioural Neurology, IRCCS Santa Lucia Foundation, Via Ardeatina 306, Rome 00179, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Bossù
authors.affiliation: Department of Clinical & Behavioural Neurology, IRCCS Santa Lucia Foundation, Rome 00179, Italy.
authors.forename: Eleonora
authors.initials: E
authors.lastname: Sacchinelli
authors.affiliation: Department of Clinical & Behavioural Neurology, IRCCS Santa Lucia Foundation, Rome 00179, Italy.
authors.forename: Maria Donata
authors.initials: MD
authors.lastname: Orfei
authors.affiliation: Department of Clinical & Behavioural Neurology, IRCCS Santa Lucia Foundation, Rome 00179, Italy.
authors.forename: Federica
authors.initials: F
authors.lastname: Piras
authors.affiliation: Department of Clinical & Behavioural Neurology, IRCCS Santa Lucia Foundation, Rome 00179, Italy.
authors.forename: Nerisa
authors.initials: N
authors.lastname: Banaj
authors.affiliation: Department of Clinical & Behavioural Neurology, IRCCS Santa Lucia Foundation, Rome 00179, Italy.|Beth K & Stuart C Yudofsky Division of Neuropsychiatry, Menninger Department of Psychiatry & Behavioral Sciences, Baylor College of Medicine, Houston, TX 77030, USA.
authors.forename: Gianfranco
authors.initials: G
authors.lastname: Spalletta
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods*; D003687:Dehydroepiandrosterone / Q000097:blood*; D019314:Dehydroepiandrosterone Sulfate / Q000097:blood; D016903:Drug Monitoring; D006801:Humans; D057230:Limit of Detection; D011280:Pregnanolone / Q000031:analogs & derivatives / Q000097:blood*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: C579439:epiallopregnanolone sulfate; D003687:Dehydroepiandrosterone; D019314:Dehydroepiandrosterone Sulfate; D011280:Pregnanolone
keywords: allopregnanolone; dehydroepiandrosterone; mass spectrometry; neurosteroids; plasma; pregnanolone