The environmental impacts of graphite exfoliation using biological synthesis were quantified using the life cycle assessment (LCA) approach as per ISO 14040-44 (ISO 14040, 2006; ISO 14044, 2006). Four steps were involved in the process (a) goal and scope definition, (b) life cycle inventory (LCI), (c) life cycle impact assessment (LCIA), and (d) life cycle interpretation. This study's primary goal is to evaluate environmental impacts of graphite exfoliation using biological synthesis. The scope of this LCA includes only the exfoliation of graphite into fewlayers graphene (gate-to-gate). The upstream life cycle stages utilization of graphene and disposal stages were excluded. The energy calculations were performed using specific heats of materials and/or the power rating of the equipment used in the synthesis process. The location of graphite exfoliation plant was considered Kharagpur, India, the source of materials and chemicals was based on the local conditions (Kolkata, India). The eastern India electricity power mix was used for energy consumption. The study's functional unit was the exfoliation of 1 ton of graphite. The functional unit was chosen to compare the feasibility of producing graphene from waste-based feedstocks from an environmental perspective at an industrial scale. The parameters and components in synthesis routes for graphite exfoliation were scaled to the industrial level and were normalized to the functional unit. The scaled-up quantities of materials and amount of energy were normalized to the functional unit, assuming that reaction rates remain unaltered at the industrial scale. Environmental impacts were evaluated using SimaPro 8.0.3 version software and Eco-invent database. IMPACT 2002+ ® method was used for the characterization of end-point impacts. Since we are aiming to study the industrial-scale exfoliation of graphite, we choose the process contributions which are most relevant to the production processes. Sensitivity analysis was performed by varying the country's electricity mix and quantity of deionized water. In the present investigation, a high-voltage electricity grid mix of the Indian geographic location was used for modeling the scenarios. For sensitivity analysis, the electricity mix of China and Norway was modeled to compare and evaluate the impacts associated based on the shift towards reduction in the coal-based electricity mix.