BACKGROUND: Stress activates the central nervous, the autonomic nervous, and the 
endocrine system. This study aimed to (1) test the usability of salivary 
cortisone in a standardized psychosocial stressor, (2) create a comprehensive 
profile of hormonal responses to determine laboratory parameters with high 
discriminatory power, and (3) analyze their association with psychometric and 
autonomic stress measures.
METHODS: Healthy young men (18-35 years) completed either the Trier Social 
Stress Test (TSST) (n = 33) or a Placebo-TSST (n = 34). Blood and saliva were 
collected at 14 time points along with state-anxiety (STAI) and heart rate. 
Serum steroids (cortisol*, cortisone*, dehydroepiandrosterone-sulfate, 
androstenedione*, progesterone*, 17-hydroxyprogesterone*, testosterone, 
estradiol*, aldosterone*), salivary cortisol* and cortisone*, copeptin*, 
adrenocorticoptropic hormone*, corticosteroid-binding globulin, and salivary 
alpha-amylase* were analyzed. We used mixed-design ANOVAs to test group 
differences, receiver operator characteristic (ROC) curve analyses to assess the 
discriminatory power of each measure, and Spearman correlation analyses to probe 
the association between measures.
RESULTS: The largest area under the ROC curve was observed in salivary cortisone 
at 20 min after the end of the TSST (AUC = 0.909 ± 0.044, p < 0.0001). 
Significant time-by-group interactions were found in the parameters marked with 
* above, indicating stress-induced increases. The peak response of salivary 
cortisone was significantly associated with those of STAI (rho = 0.477, 
p = 0.016) and heart rate (rho = 0.699, p < 0.0001) in the TSST group.
CONCLUSION: Our study found salivary cortisone to be a stress biomarker with 
high discriminatory power and significant correlations with subjective and 
autonomic stress measures. Our results can inform future stress studies of 
sampling time for different laboratory parameters.