2.6. Statistical Analyses
anxiety
As no clinical study has investigated the effect of saffron extract in response to an acute laboratory stressor on salivary cortisol response, the sample size calculation was based on the effect size of a lemon balm food supplement on salivary cortisol in healthy adults 1 h post-treatment [Demographic and lifestyle characteristics at baseline (V0) and pre-treatment values of VAS stress, VAS anxiety, salivary cortisol and cortisone collected during testing visits were compared according to the sequence using a linear mixed model (PROC MIXED). The model included sequence, period, and treatment as fixed effects and participants within sequence as the random effects. In order to confirm that participants found the MAST stressful, VAS anxiety, VAS stress, salivary cortisol, and cortisone were compared before, during, and after the MAST using the linear mixed model as above and using the time of sample collection as additional fixed factor.The incremental area under the curve (AUC) for VAS stress, VAS anxiety, salivary cortisol, and cortisone were calculated using the trapezoid rule, discarding the area beneath the pre-treatment values, considered the baseline value [All statistical tests were 2-sided, and a 