박소연 (한국과학기술연구원, 한양대학교)

최만호 (한국과학기술연구원)

박소연 (한국과학기술연구원, 한양대학교)

박승신 (서울대학교)

박민정 (고려대학교)

이준석 (한양대학교)

김정희 (서울대학교)

최만호 (한국과학기술연구원)

Although mass spectrometry is the gold standard for steroid profiling, it requires complex sample purification and technical expertise.  Since saliva is a potential alternative to blood, we simplified the LC-MS method for quantifying cortisol, cortisone, and dehydroepiandrosterone sulfate in saliva.  The assay involves protein precipitation with a deuterated internal standard solution followed by filtration.  The limit of quantification for all analytes was 0.2 – 2.0 ng/mL, with good linearity (r² > 0.97) across a calibration range of 0.1 to 100 ng/mL using 100 mL saliva.  Intra-day precision and accuracy ranged from 1.5% to 24.7% and 83.1% to 114.7%, respectively, while inter-day reproducibility ranged from 2.8% to 19.1% for precision and 81.1% to 109.1% for accuracy. Recovery rates were between 88% and 128%.  The method was then applied to salivary samples from eight patients with adrenal insufficiency and seventeen controls without adrenal dysfunctions following adrenocorticotropic hormone (ACTH) stimulation.  The results were compared with those obtained from our conventional assays in both saliva and serum samples.  Good correlations were observed with the conventional salivary assay (r = 0.92 – 0.98), and with serum concentrations (r = 0.42 – 0.67).  The salivary and serum responses to ACTH stimulation were consistent, except for cortisone, which showed a distinct increase in salivary levels.  This method offers a practical and reliable approach for assessing steroids in saliva, and may prove valuable in clinical settings where steroid levels serve as diagnostic biomarkers.