Hierapolis of Phrygia, an archaeological site in southwestern Turkey, has been a UNESCO World Heritage Site since 1988. During archaeological campaigns, 71 mortar samples from public buildings were collected, dating from the Julio-Claudian to the Middle Byzantine period. The samples were analyzed using a multi-analytical approach including polarized optical microscopy (POM), digital image analysis (DIA), X-ray powder diffraction (XRPD) and SEM–EDS to trace the raw materials and understand the evolution of mortar composition and technology over time. During the Roman period, travertine and marble were commonly used in binder production, while marble dominated in the Byzantine period. The aggregates come mainly from sands of the Lycian Nappe and Menderes Massif, with carbonate and silicate rock fragments. Variations in composition, average size and circularity suggest changes in raw material sources in both Roman and Byzantine periods. Cocciopesto mortar was used in water-related structures from the Flavian to the Severan period, but, in the Byzantine period, it also appeared in non-hydraulic contexts. Straw became a common organic additive in Byzantine renders, marking a shift from the exclusively inorganic aggregates of Roman renders. This study illustrates the evolving construction technologies and material sources used throughout the city’s history.