Analytical and Bioanalytical Chemistry

, Volume 411, Issue 6, pp 1203–1210 | Cite as

Noninvasive determination of human cortisol and dehydroepiandrosterone sulfate using liquid chromatography-tandem mass spectrometry

  • Zhimin (Tim) CaoEmail author
  • Stephanie E. Wemm
  • Liqiao Han
  • David C. Spink
  • Edelgard Wulfert
Research Paper


Liquid chromatography-tandem mass spectrometry (LC-MS/MS) for measurements of steroids in human saliva has garnered increased interest in the area of clinical psychoneuroendocrinological research. However, performance characteristics of LC-MS/MS methods for the analysis of steroids in saliva are limited. Human saliva samples were collected via passive drool. Cortisol and dehydroepiandrosterone sulfate (DHEA-S) in the samples were extracted together, resolved on a C18-A column, and analyzed using tandem mass spectrometry. The LC-MS/MS method had limits of quantitation of 0.03 and 0.06 ng/mL for DHEA-S and cortisol, respectively. Method evaluations showed coefficient variation (%CV) of inter-assay ranging 4.6–17.9% for DHEA-S and cortisol, recoveries of 102.4–109.5% for DHEA-S and 94.6–98.3% for cortisol, and assay linearity with R2 = 0.9964 for DHEA-S (1.0–25.0 ng/mL) and R2 = 0.997 (1.0–25.0 ng/mL) for cortisol. No cross contamination among samples was observed. Human saliva showed 20% and 18% ion enhancement effect for DHEA-S and cortisol assay, respectively. No interference by ten common steroids was detected. Regression analysis of method comparisons with laboratory-developed test (LDT) method revealed R2 = 0.9688 (LC-MS/MS = 0.9665 LDT-LC-MS/MS − 0.7355) for cortisol, and R2 = 0.9039 (LC-MS/MS = 1.0173 LDT-LC-MS/MS + 3.6797) for DHEA-S. Reference ranges for young adults were determined to be 0.3–5.9 ng/mL for females and 0.1–5.6 ng/mL for males for salivary cortisol, and 0.6–7.4 ng/mL for females and 0.6–10.1 ng/mL for males for salivary DHEA-S. An LC-MS/MS method for quantifying cortisol and DHEA-S in human saliva was developed and validated for clinical and psychoneuroendocrinological research that require noninvasive means of measuring these hormones.


Saliva Cortisol DHEA-S Dehydroepiandrosterone sulfate Mass spectrometry 





Dehydroepiandrosterone sulfate


Liquid chromatography-tandem mass spectrometry


Laboratory-developed test


Limit of detection


Limit of quantitation


Proficiency test



We also thank the National Natural Science Foundation of China (Grant# 81572088 & 81601861) for the support of Liqiao Han in carrying out the study at Wadsworth Center, New York State Department of Health.

Funding information

This project was supported in part by a grant (F31 DA038931) from the National Institute on Drug Abuse.

Compliance with ethical standards

The study protocol was approved by the Institutional Review Board (IRB) or Ethical Committee of the University at Albany, State University of New York. Consents were obtained from all individual participants, who voluntarily donated saliva samples.

Conflict of interest

The authors declare that they have no conflict of interest.


Use of trade names and commercial sources is for identification only and does not constitute endorsement by the Wadsworth Center of the New York State Department of Health.


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Zhimin (Tim) Cao
    • 1
    • 2
    Email author
  • Stephanie E. Wemm
    • 3
  • Liqiao Han
    • 4
  • David C. Spink
    • 1
  • Edelgard Wulfert
    • 2
  1. 1.Wadsworth CenterNew York State Department of HealthAlbanyUSA
  2. 2.University at Albany, State University of New YorkAlbanyUSA
  3. 3.Yale University School of MedicineNew HavenUSA
  4. 4.The Second Affiliated Hospital of Guangzhou University of Chinese MedicineGuangzhouChina

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