Interfaces between Mo substrate and free-standing diamond films prepared by DC arc plasma jet operated at gas recycling mode were investigated, including for the first time used and multi-time used substrate. The morphology, phase composition and bonding state of elements in the interface between substrates and diamond films were examined by optical microscopy, XRD and XPS. The profiles of carbon concentration of Mo substrates were measured by GDOES. It showed that Mo2C and MoC were formed on the first time used Mo substrate, and MoC was found on diamond films nucleation side after detachment. It suggested that MoC was peeled off from Mo substrate. The stable Mo2C on Mo substrate was formed after multi-time use of Mo substrate. However, MoC has not been found on it. The thickness of carburizing layer on the first time used Mo substrate is up to 30µm, and the carburizing layer on the multi-time used substrate is much thicker than that on the first used. The amorphous carbon in the surface of the substrate and nucleation side of diamond films was found by XPS, including for the first time used and multi-time used substrate.