The knowledge of the material composition of works of art, e.g., pigments and binding media of paint layers, can be of great importance for conservators and art historians. A number of analytical procedures can be applied for the characterization of works of art, e.g., the microscopic examination of cross sections, x-ray diffraction, etc. The routine procedures require a sample from the original and cannot be regarded as nondestructive methods as far as the object under investigation is concerned, even if the analytical procedure itself does not destroy the sample. The article describes the physical conditions needed and the technical feasibility for the application of particle induced x-ray emission (PIXE) as a multielement analysis technique, where sampling is not required. The procedure is suitable for the investigation of the elemental composition of inorganic materials, such as pigments, and can be applied to the identification of pigments in paint layers in painting, polychromed sculptures, or works of graphic art. The technique is based on the bombardment of a minute spot of the object under investigation by protons and subsequent detection of the generated characteristic x-or gamma-radiation specific for each element present. The advantages and limits of the analytical procedure are demonstrated through the investigation of a medieval panel painting. The technique allows the analysis of microdomains and leads to excellent results. The irradiation of the paint layers did not create any changes in the visual appearance of the object either during the analytical procedure itself or during the 10 months of the painting's observation. The technique therefore can be regarded as safe and strictly nondestructive.