In this study, life cycle assessment of oxygen-18 by using cryogenic distillation of oxygen is performed using SimaPro 8.3 software. Life cycle assessment is performed to understand the environmental profile and hotspots of this process in order to be used in design and development. Simulation of oxygen-18 process is executed by Hysys software, and the required inputs and outputs for inventory of life cycle were acquired. By doing life cycle assessment and considering achieved results after characterization and normalization of inventory data it has been investigated that in the majority of environmental impacts electricity consumption has a huge contribution relative to other parts of the system like liquefied oxygen production from air separation unit, required facilities for air separation and oxygen-18 units, and needed transportation. Also, among 17 impact categories investigated in ReCiPe impact assessment method, fossil depletion, climate change (human health), particulate matter formation, climate change (ecosystem), human toxicity, and metal depletion have the most contribution in entire environmental loads respectively. Furthermore, sensitivity analysis showed that changing life cycle impact assessment method fromReCiPe to IMPACT 2002+has no significant effect on acquired results and results are confident. In addition, assumption of market for depleted oxygen from heavy isotopes which is withdrawn from top of distillation columns showed some positive effects compared to first case and environmental impacts resulted from liquefied oxygen production (feed) reduced but because of huge contribution of electricity consumption compared to other sections, this positive effect has no remarkable influence on entire environmental loads of product system.