Road markings play an important role in road safety because they provide significant information to drivers about the road. For that reason, they must be replaced when they are not correctly perceived by road users. To analyse which are the main factors that affect road marking perception over time, a test section was designed in a two-lane rural highway, running actual traffic over 18 different types of markings fabricated with different combinations of drop-on materials. Chromatic coordinates, luminance and retroreflectivity of each sample were measured during 18 months in order to study its evolution over time. The results obtained show different behaviours depending on the aggregates and application method used. An increment of the durability has been observed with the use of different layers and mixtures of glass microbeads with different sizes.