Tadalafil is one of the selective phosphodiesterase type 5 inhibitors (PDE5) and serves as the active compound in drugs used for the treatment of erectile dysfunction. These PDE5 inhibitors are prescribed under medical supervision. However, cases of adulteration of dietary supplements with PDE5 inhibitors or their unapproved analogs have been reported worldwide. The presence of the PDE5 inhibitors in such supplements poses a serious health risk to consumers, particularly when combined with certain nitrate-containing drugs, as their toxic effects have not been thoroughly assessed and may result in unpredictable adverse reactions. Therefore, it is crucial to detect adulteration in these dietary supplements. However, current methods for PDE5 inhibitor detection rely on time-consuming and expensive analytical techniques, although they are sensitive. In this study, we propose an aptasensor based on ellipsometry for the detection of PDE5 inhibitors. To enhance the detection specificity for PDE5 inhibitors, we designed an aptamer with a hydrophobic pocket that incorporates a guanine base-rich region and a three-way junction. This design is particularly effective considering the poor aqueous solubility of PDE5 inhibitors. Preliminary results demonstrate that tadalafil detection in various media can be achieved within the range of 1-2000 ng/mL. The limit of detection for the active compound of tadalafil is as low as 1.82 ng/mL.Copyright © 2023. Published by Elsevier B.V.