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

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    Authors
    Wear, Joanne E
    Owen, Laura J
    Duxbury, Katherine
    Keevil, Brian G
    Affiliation
    Department of Clinical Biochemistry, Wythenshawe Hospital, Southmoor Road, Wythenshawe, Manchester M23 9LT, United Kingdom. Joanne.wear@nhs.net
    Issue Date
    2007-10-15
    
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    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.
    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
    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
    ae974a485f413a2113503eed53cd6c53
    10.1016/j.jchromb.2007.08.001
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