, Perform a saliva analysis

, Determine the participant's cortisol level by the enzyme-linked immunosorbent assay (ELISA) technique using assay kits (see Table of Materials)

, Measure the concentration related to the absorbance at 450 nm with a 96-well microplate spectrophotometer (see Table of Materials)

, Calculate the intra-assay precision with the average value of individual variability for all of the duplicates within each microplate

, Obtain concentrations of unknown samples (nmol/L) with a four-parameter nonlinear regression curve fit, following the ELISA kit protocol recommendations

, Calculate the effect sizes with a partial eta squared. NOTE: Eta squared is a statistical measure of effect size for use in ANOVA

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