A simplified method for the measurement of urinary free cortisol using LC-MS/MS.

2.50
Hdl Handle:
http://hdl.handle.net/10541/70194
Title:
A simplified method for the measurement of urinary free cortisol using LC-MS/MS.
Authors:
Wear, Joanne E; Owen, Laura J; Duxbury, Katherine; Keevil, Brian G
Abstract:
The measurement of 24 h urinary free cortisol is used in the investigation of patients with symptoms of hypercortisolism. Many different methods have been published for the measurement of cortisol, but most of these methods involve cumbersome pre-extraction of the cortisol prior to analysis. We have developed a method using in-well protein precipitation which serves to clean up the sample without requiring lengthy sample preparation. A Shimadzu SIL-HT autosampler was used to inject 50 microL of extract onto a Phenomemex Gemini C18 guard column attached to a Waters Xbridge C18 column. The eluant was introduced directly into a Waters Quattro Micro tandem mass spectrometer. The method was found to be linear up to 3448 nmol/L with a lower limit of detection of 5.3 nmol/L. Precision and accuracy were acceptable, and no interference was noted from compounds such as prednisolone or fenofibrate. This assay was compared to a previously published method, which uses solid phase extraction prior to LC-MS/MS analysis. We have developed a simplified, robust assay for the quantitation of urinary free cortisol that will increase the throughput of the assay and avoid the use of neurotoxic solvents such as dichloromethane.
Affiliation:
Department of Clinical Biochemistry, Wythenshawe Hospital, Southmoor Road, Wythenshawe, Manchester M23 9LT, United Kingdom. Joanne.wear@nhs.net
Citation:
A simplified method for the measurement of urinary free cortisol using LC-MS/MS. 2007, 858 (1-2):27-31 J. Chromatogr. B
Journal:
Journal of Chromatography. B
Issue Date:
15-Oct-2007
URI:
http://hdl.handle.net/10541/70194
DOI:
10.1016/j.jchromb.2007.08.001
PubMed ID:
17698425
Type:
Article
Language:
en
ISSN:
1570-0232
Appears in Collections:
All Christie Publications

Full metadata record

DC FieldValue Language
dc.contributor.authorWear, Joanne E-
dc.contributor.authorOwen, Laura J-
dc.contributor.authorDuxbury, Katherine-
dc.contributor.authorKeevil, Brian G-
dc.date.accessioned2009-06-11T13:12:56Z-
dc.date.available2009-06-11T13:12:56Z-
dc.date.issued2007-10-15-
dc.identifier.citationA simplified method for the measurement of urinary free cortisol using LC-MS/MS. 2007, 858 (1-2):27-31 J. Chromatogr. Ben
dc.identifier.issn1570-0232-
dc.identifier.pmid17698425-
dc.identifier.doi10.1016/j.jchromb.2007.08.001-
dc.identifier.urihttp://hdl.handle.net/10541/70194-
dc.description.abstractThe measurement of 24 h urinary free cortisol is used in the investigation of patients with symptoms of hypercortisolism. Many different methods have been published for the measurement of cortisol, but most of these methods involve cumbersome pre-extraction of the cortisol prior to analysis. We have developed a method using in-well protein precipitation which serves to clean up the sample without requiring lengthy sample preparation. A Shimadzu SIL-HT autosampler was used to inject 50 microL of extract onto a Phenomemex Gemini C18 guard column attached to a Waters Xbridge C18 column. The eluant was introduced directly into a Waters Quattro Micro tandem mass spectrometer. The method was found to be linear up to 3448 nmol/L with a lower limit of detection of 5.3 nmol/L. Precision and accuracy were acceptable, and no interference was noted from compounds such as prednisolone or fenofibrate. This assay was compared to a previously published method, which uses solid phase extraction prior to LC-MS/MS analysis. We have developed a simplified, robust assay for the quantitation of urinary free cortisol that will increase the throughput of the assay and avoid the use of neurotoxic solvents such as dichloromethane.en
dc.language.isoenen
dc.subject.meshChromatography, Liquid-
dc.subject.meshHumans-
dc.subject.meshHydrocortisone-
dc.subject.meshReproducibility of Results-
dc.subject.meshTandem Mass Spectrometry-
dc.titleA simplified method for the measurement of urinary free cortisol using LC-MS/MS.en
dc.typeArticleen
dc.contributor.departmentDepartment of Clinical Biochemistry, Wythenshawe Hospital, Southmoor Road, Wythenshawe, Manchester M23 9LT, United Kingdom. Joanne.wear@nhs.neten
dc.identifier.journalJournal of Chromatography. Ben
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