This article presents a comprehensive analysis of the polychrome paintings on the Flower Peking Opera Theatre in Bozhou, Anhui Province, China. A multi-technique approach was employed, including polarized light microscopy (PLM), X-ray fluorescence (XRF), micro-Raman spectroscopy, X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy with energy-dispersive spectrometry (SEM-EDS), and Herzberg staining to determine the composition and methodologies involved in the formation of the pigment layer, the white primer, and the ground layer. The analysis identified cinnabar (red), both artificial ultramarine and Prussian blue (blue), a mixture of barite and gypsum (white), a mixture of chromite and Prussian blue (green), and carbon black (black) in the pigment layer. The ground layer was found to consist of clay and plant fibers (cotton and hemp), while the white prime layer was primarily composed of barite and gypsum. This research provides insights for future conservation and restoration efforts.