The painting materials in the Islamic monuments in Cairo were variable and distinguished. The accurate identification and characterization of painting materials in an art object or in a paint layer is certainly an important step in the history of art, technology and conservation. The main objective of the present paper is to examine and investigate the different painting materials (pigments, gilding, binding media and painting ground) employed in the decorated ceilings of Madrasa of Al-Ashraf Bersbay (Mamluk Jarkasy period). X-ray diffraction (X.R.D.), scanning electron microscope SEM (EDAX), light optical microscope (LOM) and fourier transform infrared spectroscopy (FTIR), together with determination of physical properties of the support (water content and density) and microbiological study, adapted for analysis, investigation and evaluation of the current status. The study revealed the severe damage of painting materials. Gypsum, ultramarine, smalt, prussian blue, red ochre, emerald green, red lead (minium) and gold leaves were identified in El-Ashraf Bersbay Madrasa decoration painting