, Volume 59, Issue 3, pp 520–528 | Cite as

Characterization of the serum and salivary cortisol response to the intravenous 250 µg ACTH1-24 stimulation test

  • Brendan J. Nolan
  • Jane Sorbello
  • Nigel Brown
  • Goce Dimeski
  • Warrick J. InderEmail author
Original Article



The ACTH1–24 stimulation test is commonly used to assess the hypothalamic-pituitary-adrenal (HPA) axis. Given variations in CBG concentration and binding affinity, serum total cortisol may misclassify some patients. Salivary cortisol correlates well with serum free cortisol but is easier to measure and widely available in commercial laboratories. The aim of this study was to investigate the utility of measuring salivary cortisol during the ACTH1–24 stimulation test.

Design and methods

Case–control study in a clinical research facility. Eighty-seven patients with suspected cortisol deficiency, twenty-four healthy controls, and ten healthy women on the oral contraceptive (OC) underwent an intravenous 250 µg ACTH1–24 stimulation test. Concordance of ACTH1–24 stimulated serum and salivary cortisol was evaluated.


There was a significant difference in serum cortisol between the healthy volunteers and the women on the OC (P < 0.001) but no difference in salivary cortisol. The lower limit of the reference interval for salivary cortisol at 60 min was 26 nmol/L. 27/89 (30%) of tests with suspected HPA axis disorder failed the 60 min serum cortisol cut-off of 500 nmol/L. Of these, 24/27 (89%) had a salivary cortisol of <26 nmol/L. In contrast, 12/19 (63%) tests and 5/43 (12%) tests where the 60 min serum cortisol was 500–599 and ≥600 nmol/L, respectively had a salivary cortisol of <26 nmol/L.


Salivary cortisol provides additional diagnostic value during the 250 µg ACTH1–24 stimulation test in patients with proven or suspected alterations in CBG and potentially those with a borderline 60 min serum cortisol 500–599 nmol/L.


Salivary cortisol ACTH1–24 stimulation test Oral contraceptive Adrenal insufficiency 



This study was funded by a grant from the Study Education Research Committee, Pathology Queensland, Brisbane, Australia.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in the study.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Diabetes and EndocrinologyPrincess Alexandra HospitalWoolloongabbaAustralia
  2. 2.Faculty of MedicineUniversity of QueenslandBrisbaneAustralia
  3. 3.Department of Chemical PathologyPathology QueenslandBrisbaneAustralia

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