Simultaneous measurement of cortisol and cortisone in human saliva using liquid chromatography-tandem mass spectrometry: application in basal and stimulated conditions.

2.50
Hdl Handle:
http://hdl.handle.net/10541/109253
Title:
Simultaneous measurement of cortisol and cortisone in human saliva using liquid chromatography-tandem mass spectrometry: application in basal and stimulated conditions.
Authors:
Perogamvros, Ilias; Owen, Laura J; Newell-Price, John; Ray, David W; Trainer, Peter J; Keevil, Brian G
Abstract:
Immunoassays used for the measurement of salivary cortisol are limited by variable interference from cortisone. Salivary cortisone is a consequence of the salivary glands expressing 11beta-hydroxysteroid dehydrogenase type 2 (11beta-HSD2) which converts cortisol to cortisone. We report a combined salivary cortisol and cortisone (SalF and SalE respectively) liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay to address the cortisone cross-reactivity in cortisol immunoassays and as a tool to study 11beta-HSD2 activity. The method was linear up to 400 nmol/L for SalF and 200 nmol/L for SalE and the lower limits of quantitation were 0.39 nmol/L (SalF) and 0.78 nmol/L (SalE). No evidence of ion suppression was found and precision, accuracy and recovery were within internationally accepted limits. No interference was identified from 13 structurally related steroids. SalF, SalE and SalF/SalE were significantly greater in the morning than at bed-time and following stimulation of the adrenal glands. As serum cortisol increased, an exponential rise was observed in SalF and a linear increase in SalE which reached a plateau at higher SalF concentrations. We have developed a novel, robust LC-MS/MS assay for the combined measurement of SalF and SalE. Our results confirm the 11beta-HSD2 activity of the salivary glands resulting in high SalE concentrations and the enzyme saturation at high substrate concentrations. This method can be used as a simple, non-invasive and highly specific tool to assess the value of salivary cortisol as a surrogate for free serum cortisol and as a potential novel way to assess 11beta-HSD2 activity.
Affiliation:
Department of Endocrinology, The Christie, Manchester, UK. ilias.perogamvros@nhs.net
Citation:
Simultaneous measurement of cortisol and cortisone in human saliva using liquid chromatography-tandem mass spectrometry: application in basal and stimulated conditions. 2009, 877 (29):3771-5 J. Chromatogr. B Analyt. Technol. Biomed. Life Sci.
Journal:
Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Issue Date:
1-Nov-2009
URI:
http://hdl.handle.net/10541/109253
DOI:
10.1016/j.jchromb.2009.09.014
PubMed ID:
19783236
Type:
Article
Language:
en
ISSN:
1873-376X
Appears in Collections:
All Christie Publications ; Endocrinology

Full metadata record

DC FieldValue Language
dc.contributor.authorPerogamvros, Iliasen
dc.contributor.authorOwen, Laura Jen
dc.contributor.authorNewell-Price, Johnen
dc.contributor.authorRay, David Wen
dc.contributor.authorTrainer, Peter Jen
dc.contributor.authorKeevil, Brian Gen
dc.date.accessioned2010-08-07T10:41:52Z-
dc.date.available2010-08-07T10:41:52Z-
dc.date.issued2009-11-01-
dc.identifier.citationSimultaneous measurement of cortisol and cortisone in human saliva using liquid chromatography-tandem mass spectrometry: application in basal and stimulated conditions. 2009, 877 (29):3771-5 J. Chromatogr. B Analyt. Technol. Biomed. Life Sci.en
dc.identifier.issn1873-376X-
dc.identifier.pmid19783236-
dc.identifier.doi10.1016/j.jchromb.2009.09.014-
dc.identifier.urihttp://hdl.handle.net/10541/109253-
dc.description.abstractImmunoassays used for the measurement of salivary cortisol are limited by variable interference from cortisone. Salivary cortisone is a consequence of the salivary glands expressing 11beta-hydroxysteroid dehydrogenase type 2 (11beta-HSD2) which converts cortisol to cortisone. We report a combined salivary cortisol and cortisone (SalF and SalE respectively) liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay to address the cortisone cross-reactivity in cortisol immunoassays and as a tool to study 11beta-HSD2 activity. The method was linear up to 400 nmol/L for SalF and 200 nmol/L for SalE and the lower limits of quantitation were 0.39 nmol/L (SalF) and 0.78 nmol/L (SalE). No evidence of ion suppression was found and precision, accuracy and recovery were within internationally accepted limits. No interference was identified from 13 structurally related steroids. SalF, SalE and SalF/SalE were significantly greater in the morning than at bed-time and following stimulation of the adrenal glands. As serum cortisol increased, an exponential rise was observed in SalF and a linear increase in SalE which reached a plateau at higher SalF concentrations. We have developed a novel, robust LC-MS/MS assay for the combined measurement of SalF and SalE. Our results confirm the 11beta-HSD2 activity of the salivary glands resulting in high SalE concentrations and the enzyme saturation at high substrate concentrations. This method can be used as a simple, non-invasive and highly specific tool to assess the value of salivary cortisol as a surrogate for free serum cortisol and as a potential novel way to assess 11beta-HSD2 activity.en
dc.language.isoenen
dc.subject.meshChromatography, Liquid-
dc.subject.meshCortisone-
dc.subject.meshHumans-
dc.subject.meshHydrocortisone-
dc.subject.meshReproducibility of Results-
dc.subject.meshSaliva-
dc.subject.meshTandem Mass Spectrometry-
dc.titleSimultaneous measurement of cortisol and cortisone in human saliva using liquid chromatography-tandem mass spectrometry: application in basal and stimulated conditions.en
dc.typeArticleen
dc.contributor.departmentDepartment of Endocrinology, The Christie, Manchester, UK. ilias.perogamvros@nhs.neten
dc.identifier.journalJournal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciencesen
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