It was expected that replacing buffer with artificial saliva would give a similar pattern as presented above. However, a counterintuitive pattern was obtained, as shown by the inverse trend showing heightened TO fluorescence. Based on the data outlined in figure , it was assumed that elevated hydrocortisone levels in the sample would trigger an increased release of TO from the cortisol aptamer, leading to a reduction in fluorescence. However, results obtained when substituting the buffer with saliva demonstrated an increase in TO fluorescence with rising hydrocortisone concentrations from 9.96% at 0.00 µM to -2.03% at 0.50 µM of hydrocortisone (figure ). This heightened fluorescence implies that artificial saliva has a discernible impact on TO fluorescence in comparison to the HS Tris buffer.