Synthesis of 3,5-di(6-mercaptohexyloxy)toluene (DMHT) was achieved starting from orcinol. A molecularly imprinted self-assembled monolayer (SAM) was fabricated by forming an SAM with both DMHT and the template molecule (cholesterol), followed by removal of the template molecules by solvent extraction. These imprinted SAMs were capable of discriminating the imprinted substrate. The stability of the imprinted SAMs was estimated precisely using a quartz crystal microbalance. In comparison to the results used monothiol compounds, the duration of the sensing capability of this imprinted SAM was improved greatly. The two thiol functional groups in DMHT clearly had a significant effect on the duration.