In this study, Zeugma wall paintings and related materials including sediment and stone/rock had been examined. Samples were examined to get their raw material characteristics, mineralogical and chemical compositions, and microstructural properties using various analytical techniques. Sediments were calcereous soils of Eocene. Rock types of stones were mainly limestone which had 3 subgroups: micritic, biomicritic and recrytallised micritic limestones. The source of the limestones should be from the local formation. Binder of mortar samples were lime, aggregate materials come from the river deposites of Euphrates. Mural painting samples have different layers; pigment layer at the top, intonaco layer underneat, and single or double rather coarse grained arriccio layer(s) below intonaco layer. The mural painting technique was fresco technique. Calcite was common mineral identified for all pigments of different colours. The sources of white, black and green coloured pigments were found as vaterite, graphite and malachite respectively. The sources of yellow coloured pigments were identified as ankerite, siderite and goethite. The red colours were identified as hematite, jasper and red earth/ochre. Jasper and vaterite, jasper and calcite, red earth/ochre and calcite, and hematite were the colour forming minerals of pink coloured pigments.