This study applied multiple scientific approaches to establish the significance of an old work of art, Red Guitar, by examining its historical origin and the color materials used in its creation. Additionally, the study provides thus far unknown pieces of Olga Picasso’s family history to be added to her biography. Scientific approaches included digital X-ray radiography, X-ray fluorescence spectroscopy, infrared Fourier transform spectroscopy, Raman spectroscopy, and elemental thermal conductivity analysis. This combination of techniques provided a broad confirmation as to when the painting was created. The work includes colors (white, black, blue, yellow, green, red, and brown/red) and prevalent use of lead- and iron-based historic pigments—chrome yellow, yellow ochre, and red ochre. It also documents the use of unconventional materials, such as the colorant, Pigment Red 4, and nitrocellulose. This investigation led to the conclusion that the artwork, Red Guitar, is authentic and in accordance with Picasso’s work during the first two decades of the 20th century.