The interior of Orthodox churches is entirely decorated with paintings, icons, and frescoes, to help create a special environment for the prayers and visitors. The paintings have religious, esthetic, and historical value, being created in the Byzantine tradition and following recommendations on style and materials according to old church canons. Particular importance is attributed to the Iconostasis decorated with icons on several registers, as well as to polychrome wooden ornaments and imperial doors. This work presents the scientific investigation results of three royal icons, theoretically dating to the 19th century, from the Iconostasis of the Runcu church, an important 19th-century Romanian historical monument, aiming to answer questions regarding their age and constituent materials, especially what were the mineral pigments used in the color palettes for their painting and if there were any old repainting interventions, in order to assist their restoration and conservation. Methods of characterization employed were optical microscopy, SEM-EDX spectrometry, ATR/FTIR spectroscopy, and X-ray fluorescence spectrometry. Results obtained allowed us to date, attribute, and evaluate the conservation state of the Royal icons; to our knowledge, this study is among the very few research studies carried out so far on the religious heritage conservation field in Romania.