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Reference ranges of late-night salivary cortisol and cortisone measured by LC–MS/MS and accuracy for the diagnosis of Cushing’s syndrome

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Abstract

Purpose

International guidelines recommend salivary cortisol for the diagnosis of Cushing’s syndrome. Despite mass spectrometry-based assays are considered the analytical gold-standard, there is still the need to define reference intervals and diagnostic accuracy of such methodology.

Methods

100 healthy volunteers and 50 consecutive patients were enrolled to compare LC–MS/MS and electrochemiluminescence assay for the determination of late-night salivary cortisol and cortisone. Moreover, we aimed to determine reference intervals of salivary steroids in a population of healthy individuals and diagnostic accuracy in patients with suspected hypercortisolism and in a population including also healthy individuals.

Results

Method comparison highlighted a positive bias (51.8%) of immunoassay over LC–MS/MS. Reference intervals of salivary cortisol (0.17–0.97 µg/L), cortisone (0.84–4.85 µg/L) and ratio (0.08–0.30) were obtained. The most accurate thresholds of salivary cortisol for the diagnosis of hypercortisolism were 1.15 µg/L in the population with suspected hypercortisolism (AUC 1) and 1.30 µg/L in the population including also healthy individuals (AUC 1). Cut-off values of salivary cortisone (7.23 µg/L; Se 92.9%, Sp 97.2%, AUC 0.960 and Se 92.9%, Sp 99.1%, AUC 0.985 in suspected hypercortisolism and in overall population, respectively) and cortisol-to-cortisone ratio (0.20; Se 85.7%, Sp 80.6%, AUC 0.820 and Se 85.7%, Sp 85.5%, AUC 0.855 in suspected hypercortisolism and in overall population, respectively) were accurate and similar in both populations.

Conclusion

LC–MS/MS is the most accurate analytical platform for measuring salivary steroids. Obtained reference intervals are coherent with previously published data and diagnostic accuracy for diagnosis of overt hypercortisolism proved highly satisfactory.

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Funding

The study was supported by a grant from the University of Turin. The funding sources had no role in the study design, data collection and analysis, manuscript preparation, or decision to submit the paper for publication.

Author information

Author notes

  1. F. Ponzetto, F. Settanni and M. Parasiliti-Caprino are contributed equally to this work.

Authors and Affiliations

  1. Endocrinology, Diabetes and Metabolism, Department of Medical Sciences, City of Health and Science University Hospital, University of Turin, Corso Dogliotti 14, 10126, Turin, Italy

    F. Ponzetto, F. Settanni, M. Parasiliti-Caprino, F. Rumbolo & E. Ghigo

  2. Clinical Biochemistry Laboratory, City of Health and Science University Hospital, Turin, Italy

    F. Settanni, F. Rumbolo, A. Nonnato, M. Ricciardo, G. Priolo, S. Vitali & G. Mengozzi

  3. Department of Chemistry, University of Turin, Turin, Italy

    E. Amante & L. Anfossi

  4. Oncologic Endocrinology, Department of Medical Sciences, City of Health and Science University Hospital, University of Turin, Turin, Italy

    E. Arvat

  5. Department of Biological and Clinical Sciences, University of Turin, Turin, Italy

    R. Giordano

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  1. F. Ponzetto
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Contributions

F.S., F.P. had the idea and, together with M.P.C., performed the data analysis and the manuscript writing. F.R., A.N., M.R., E.A., G.P. and S.V. collected the biomaterial and performed the analyses for each patient. L.A. verified the analytical methods and supervised the manuscript drafting. R.G. and G.M. were in charge for overall direction and planning. E.A. and E.G. gave the needed encouragement and support to investigate and supervised the findings of this work. All authors discussed the results and contributed to the final manuscript.

Corresponding author

Correspondence to M. Parasiliti-Caprino.

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The study protocol was approved by the Institutional Ethics Committee.

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Written informed consent was obtained from all enrolled patients.

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Ponzetto, F., Settanni, F., Parasiliti-Caprino, M. et al. Reference ranges of late-night salivary cortisol and cortisone measured by LC–MS/MS and accuracy for the diagnosis of Cushing’s syndrome. J Endocrinol Invest 43, 1797–1806 (2020). https://doi.org/10.1007/s40618-020-01388-1

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