The ability of the pyrimidine derivatives to form numerous complexes and supramolecular assemblies makes them suitable for the construction of new functional surfaces. Therefore, in this paper, the adsorption behavior of 4-hydroxy-6-(trifluoromethyl)pyrimidine-2-thiol (HTPT) on a Au(111) surface has been investigated using electrochemical scanning tunneling microscopy (EC-STM). High-resolution imaging revealed that the HTPT molecules organize on a gold surface producing a highly ordered monolayer consistent with a (4 x root 3)R-30 degrees superstructure. It has been observed that the arrangement of the molecules, as well as their orientation with respect to the substrate, remains stable over a relatively broad potential range from -0.40 to 0.55 V. It has been demonstrated that the presence of the functional groups attached to the aromatic ring affects the final structure of the HTPT adlayer, giving rise to the formation of the assembly with a uniform orientation of the molecules oil the substrate.