In the previous paper (Part I), the colorimetry and interferometric microscopy measurements on modern gold leaf models have revealed that the visual appearance of a gilded surface, both burnished and unburnished, depends strongly on the substrate type, surface roughness and texture, but not on the colour of the substrate. In this second part, we investigate the materials compositions and technical specifications of medieval gold leaf through combining literature sources and materials analysis such as scanning electron microscopy coupled with energy dispersive X-ray analysis (SEM–EDX) on samples taken from gilded wooden sculptures. Our study shows that the late medieval gold leaf has a high purity of about 23.7 carat and has an average thickness of 160 nm (with a peak value of 138 nm), purer and thicker than the modern gold leaves studies in Part I. Supportive Rutherford backscattering spectrometry (RBS) measurements on gilded models confirms the accuracy and reliability of the SEM–EDX observations on the medieval gold leaf samples. We additionally present observations of a rarely recorded special variant of medieval gold leaf—“fine reinforced gold leaf”. Combined with the findings from Part I, we conclude that light penetrating the medieval gold leaf and reflected from the gilding substrate could not be a significant, or even perceptible contribution to the visual appearance of the gilding. We argue that the misconception surrounding the correlation between the substrate colour and the gilded surface appearance can be attributed to the historical development of gilding and polychromy technologies.