Methods that unambiguously prove microbial plastic degradation and allow for quantification of degradation rates are necessary to constrain the influence of microbial degradation on the marine plastic budget. We developed an assay based on stable isotope tracer techniques to determine microbial plastic mineralization rates in liquid medium on a lab scale. For the experiments, ¹³C-labeled polyethylene (¹³C-PE) particles (irradiated with UV-light to mimic exposure of floating plasticto sunlight) were incubated in liquid medium with Rhodococcus ruber as a model organism for proof of principle. The transfer of ¹³C from ¹³C-PE into the gaseous and dissolved CO₂ pools translated to microbially mediated mineralization rates of up to 1.2 % yr⁻¹ of the added PE. After incubation, we also found highly ¹³C-enriched membrane fatty acids of R. ruber including compounds involved in cellular stress responses. We demonstrated that isotope tracer techniques are a valuable tool to detect and quantify microbial plastic degradation.