Diffuse reflectance spectroscopy (DRS), laser-Raman spectrometry (LRS) and energy dispersive X-ray fluorescence (EDXRF) were employed for the characterization of pigments on a portrait of Chinese Taoism figure on paper. The results showed clearly cinnabar, emerald green, ultramarine, orpiment, carbon black, and lead white were mainly used. To determine whether the ultramarine used was synthetic or natural, its crystal structure was characterized by X-ray diffraction (XRD). The results showed the blue pigment was synthetic ultramarine. Ultramarine blue and emerald green were synthesized in 1828 and 1814, respectively and introduced to China in the late Qing Dynasty. It was concluded that the portrait was made no earlier than the late Qing Dynasty. This study shows that DRS, LRS, and EDXRF are ideal and powerful tools for non-destructive identification of pigments on fragile paper-based cultural materials. The technique has broad application of analysis of cultural relics.