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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Greaves, Ronda / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter

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Volume 44, Issue 12


Measurement of late-night salivary cortisol with an automated immunoassay system

Michael Vogeser / Jürgen Durner / Ewald Seliger
  • Department of Obstetrics and Reproductive Medicine, Hospital of the Martin Luther University, Halle, Germany
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Christoph Auernhammer
Published Online: 2006-12-13 | DOI: https://doi.org/10.1515/CCLM.2006.244


Background: Measurement of late-night salivary cortisol concentrations is increasingly used as a screening test in suspected Cushing's syndrome. Cortisol concentrations are typically extremely low in late-night samples and discordant assay-specific reference ranges have been reported. Therefore, the aim of our study was to assess the analytical performance of the first automated cortisol immunoassay specified for salivary measurements and to establish late-night sampling reference-range data for this test.

Methods: Salivary cortisol was measured using the Roche Cobas Cortisol assay (Roche Diagnostics). Five salivary pools in different concentration ranges were used to assess the inter-assay imprecision of this test in a two-centre evaluation protocol including two reagent lots. Linearity was tested by serial dilution. Salivary samples were obtained at 23:00h from 100 apparently healthy volunteers using a commercially available salivary sampling device (Salivette, Sarstedt). A subset of 20 samples was used for method comparison with isotope dilution liquid chromatography-tandem mass spectrometry.

Results: Inter-assay coefficients of variation (n=20) between 11.6% and 40.4% were found for mean cortisol concentrations between 12.9 and 2.6nmol/L, with an estimated functional sensitivity of approximately 5.0nmol/L. The test also gave linear results in the lowest concentration range between 1.0 and 8.3nmol/L. Mean late-night salivary cortisol of 5.0nmol/L was found for healthy individuals; the absolute range was 1.4–16.7nmol/L, and the 95th percentile was 8.9nmol/L. Substantially lower concentrations were found with isotope dilution LC-MS/MS compared to immunoassay results (mean concentrations 1.8 and 4.4nmol/L, respectively).

Conclusions: The automated assay investigated was found to offer acceptable analytical performance in the very low concentration range required for late-night salivary cortisol, despite a very short turn-around time. Using this assay, late-night salivary cortisol concentrations below 8.9nmol/L are typically found in healthy volunteers.

Clin Chem Lab Med 2006;44:1441–5.

Keywords: immunoassay; late-night salivary cortisol; liquid chromatography-tandem mass spectrometry (LC-MS/MS); reference range


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About the article

Corresponding author: Michael Vogeser, MD, Institute of Clinical Chemistry, Hospital of the University of Munich, Marchioninistraße 15, 81377 Munich, Germany Phone: +49-89-70953221, Fax: +49-89-70953240,

Received: 2006-04-28

Accepted: 2006-09-23

Published Online: 2006-12-13

Published in Print: 2006-12-01

Citation Information: Clinical Chemistry and Laboratory Medicine (CCLM), Volume 44, Issue 12, Pages 1441–1445, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/CCLM.2006.244.

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