The pictorial materials used in the decoration of the front page of a Renaissance Portuguese Charter, 1512, were studied. The front page is an illuminated manuscript incorporating a decorative border, and begins with a gilded initial; it presents iron gall calligraphy and also a red decorated initial A. The colours, applied on parchment, are orange, red, green, blue, gold and a reddish brown. They were applied with a common binding medium, a vegetal gum. With the exception of the brownish red, all the colorants were of inorganic origin, synthetic or mineral: vermilion, red lead, azurite and malachite. In the synthetic colours, fillers such as calcium carbonate and lead white were added. The reddish brown used in the background of the main initial is an organic dye. Gold was used in the main initial as well as in some details in the decorative border. The gold was applied on a substrate of gypsum and lead white in a proteinaceous medium. The materials were characterized by non-destructive techniques, in situ (microEDXRF, UV-VIS emission fluorescence, colorimetry), or, when needed, by micro-sampling (microFTIR), and the results were compared to what is described in medieval treatises on the Art of illumination, as De Arte Illuminandi and The book on how one makes colours. (c) 2007 Elsevier Masson SAS. All rights reserved.