The paper presents our results concerning a complex investigation by reflectance spectroscopy (RS) and Electron Paramagnetic Resonance (EPR) of the defects induced by gamma irradiation in 22 different historical pigments. Gamma irradiation is used to destroy microflora and insects which are involved in biodeterioration processes of art works such as paintings. At the same time, it can induce defects i.e. color centers which are likely to modify the original painting colors by altering the embedded pigments. Accordingly, RS was used to quantify, by means of CIELAB color space, the contribution of irradiation defects to the pigments color changes, while EPR spectroscopy, in view of paramagnetic properties of color centers, was used to confirm their presence after irradiation. Our investigations showed that, excepting marble dust whose color alteration was still observable after 3months, color changes induced by irradiation in all other pigments disappeared after about 30–40days. In addition, RS as well as EPR measurements suggest that color changes are related to irradiation color centers, this finding being confirmed by the coincidence, within experimental uncertainties, of the half-life time color changes as obtained by these methods.