Here, a systematic study in order to assess the potential of THz time domain reflectometry for measuring the thicknesses of overpaint layers applied on original gilded surfaces was carried out. The work is part of a thorough characterization campaign, which is going on at the Rijksmuseum for addressing the conservation problems of a set of 19th century gilded picture frames on which heavy coatings were applied in previous undocumented restoration interventions. To perform such non-invasive thickness measurements, an analytical protocol based on Gaussian fits of the THz pulse-echo temporal profiles was optimized through the preparation of suitable technical samples and the comparison with direct thickness measurements. Finally, the methodology was validated by characterizing the microstratigraphy of an original sculptural element from a gilded picture frame in the Rijksmuseum collection. The results achieved show the effectiveness of the present approach in revealing multi-layered dielectric microstructures with a spatial resolution of about 30 µm when using a spectral range up to 1.5 THz.