The mainly use of infrared spectroscopy and colorimetry allowed the characterization of the pigments and compounds of the wall paintings from the Monastery of San Isidoro del Campo in Seville, dated on the 15th century. Calcite, quartz and albite were detected in the ground mortar layers. As pigments, red ochre, yellow ochre, paratacamite, atacamite, azurite and carbon black were used, and also lime as the binder, therefore the fresco technique was employed. Both FTIR-ATR and FTIR (in transmission) experiments were performed. Comparison between materials used in some of the wall paintings from Seville in this period was performed. The article revealed important information about wall paintings within a historical context related to the 15th century Catholic Reformation.