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

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Abstract

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.

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Abbreviations

ACN:

Acetonitrile

DHEA-S:

Dehydroepiandrosterone sulfate

LC-MS/MS:

Liquid chromatography-tandem mass spectrometry

LDT:

Laboratory-developed test

LoD:

Limit of detection

LoQ:

Limit of quantitation

PT:

Proficiency test

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Acknowledgements

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

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

Author information

Authors and Affiliations

  1. Wadsworth Center, New York State Department of Health, Empire State Plaza, PO Box 509, Albany, NY, 12201-0509, USA

    Zhimin (Tim) Cao & David C. Spink

  2. University at Albany, State University of New York, 1400 Washington Avenue, Albany, NY, 12222, USA

    Zhimin (Tim) Cao & Edelgard Wulfert

  3. Yale University School of Medicine, 333 Cedar St, New Haven, CT, 06510, USA

    Stephanie E. Wemm

  4. The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, 111 Dade Road, Guangzhou, 510120, China

    Liqiao Han

Authors
  1. Zhimin (Tim) Cao
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  2. Stephanie E. Wemm
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  3. Liqiao Han
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Corresponding author

Correspondence to Zhimin (Tim) Cao.

Ethics declarations

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.

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The authors declare that they have no conflict of interest.

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Cao, Z., Wemm, S.E., Han, L. et al. Noninvasive determination of human cortisol and dehydroepiandrosterone sulfate using liquid chromatography-tandem mass spectrometry. Anal Bioanal Chem 411, 1203–1210 (2019). https://doi.org/10.1007/s00216-018-1549-x

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  • DOI: https://doi.org/10.1007/s00216-018-1549-x

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