2,3-Butanediol (2,3-BDO), which is used in various fields such as cosmetics and fertilizers, is a high valueadded substance and the demand for it is gradually increasing. 2,3-BDO produced from the fermentation of microorganisms not only contains by-products of fermentation, but also varies greatly in feed composition depending on fermentation conditions, so it is difficult to efficiently operate the separation process to reach the target purity of the product. Therefore, in this study, through dynamic optimization of the bio-based 2,3-BDO distillation process, the optimal control route was explored to control the 2,3-BDO concentration of the bottom product to 99 wt% or more, when feed concentration changes. Steady and dynamic state process simulation, proportional integral (PI) controller design, and dynamic optimization were sequentially performed. As a result, the error between the 2,3-BDO concentration and the set point of the bottom product was reduced by 75.2%.