Abstract: The complex structural composition of biomass resulted in low raw material utilization and high production costs. The separation of biomass fractionation was the premise to achieve efficient utilization of all components. Based on the characteristics of biomass component separation using chemical methods, it could be categorized into three categories: cellulose-first, lignin-first, and hemicellulose-first. The characteristics of different component prioritization strategies and the main methods included in each strategy were introduced in this paper. The latest advancements in various methods under different strategies were emphatically reviewed. Moreover, the strengths and weaknesses of each method, as well as their effects on the separation of different components, were analyzed. Additionally, the research focus on the future optimization of efficient biomass fractionation technologies and how to drive the development of bio-refining was prospected.