The results derived from the examination of hydrocortisone within artificial saliva exhibited a contrasting pattern (figure ) when contrasted with the outcomes observed for hydrocortisone in HS Tris buffer (figure ). A plausible explanation for these unexpected findings could be attributed to the presence of mucins within the artificial saliva, a component that also exists in authentic human saliva . These glycoproteins characterised by a peptide backbone and intricate carbohydrate side chains. These attributes contribute to the formation of a random coil structure, consequently elevating the viscosity of saliva . While existing literature lacks insights into the impact of mucins on TO fluorescence, this study aims to draw parallels between the structure of mucins and the assembly of amyloid fibrils, which are organised protein aggregates. previously illustrated that the aggregation of amyloid fibrils triggers the activation of TO's fluorescent signal. In the context of this study, it is conjectured that the amplified fluorescence of TO, following the addition of artificial saliva, can be attributed to constraints imposed upon the torsional motion around the monomethine bridge connecting the two ring components. This constraint is presumed to occur due to TO becoming entrapped within the mucin structure. Consequently, the presence of mucins within saliva contributes to an augmentation in TO fluorescence. However, this augmentation introduces complexity in the current approach to cortisol determination in saliva, as the signal associated with the hydrocortisone-aptamer interaction becomes entwined with the signal originating from TO's interaction with mucins and is thus not detected during measurments.