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The present work was aimed at characterising the materials and assessing the state of conservation of the 'Adorazione dei Magi', a wooden panel painted by Bartolo di Fredi in the second half of fourteenth century. To this goal, innovative non-destructive investigation techniques such as portable Raman spectroscopy, visible reflectance spectroscopy and unilateral nuclear magnetic resonance, were used in situ in order to achieve surface and bulk compositional data. The former allowed the identification of the artist's palette including cinnabar, red ochre, minium, carbon black, lead white, ultramarine blue, malachite and phthalocyanine green, while the latter provided information on the stratigraphic structure and state of conservation of the wooden panel.