Molybdenum oxide nanomaterials have recently attracted widespread attention for their unique optical properties and catalytic performance. However, until now, there is little literature on the application of photoluminescent molybdenum oxide nanomaterials in biological and pharmaceutical sensing. Herein, photoluminescent molybdenum oxide quantum dots (MoO x QDs) were synthesized via a facile method, and then, the synthesized MoO x QDs were further applied as a new type of photoluminescent probe to design a new off-on sensor for captopril (Cap) determination on the basis of the fact that the quenched photoluminescence of MoO x QDs by Cu2+ was restored with Cap through specific interaction between the thiol group of Cap and Cu2+. Under optimal conditions, the restored photoluminescence intensity showed a good linear relationship with the content of Cap, ranging from 1.0 to 150.0 μM, with a limit of detection of 0.51 μM (3σ/k). Additionally, the content of Cap in pharmaceutical samples was successfully detected with the newly developed off-on sensor, and the recoveries were 99.4-101.7%, which suggest that the present off-on sensor has a high accuracy.