The growing global concern for environmental preservation and the demand for sustainable technologies have stimulated the development of biotechnological processes that enable the production of energy and chemicals in an environmentally responsible manner. Among these, fermentation from lignocellulosic biomass has gained prominence as a biological alternative to achieve these goals. However, the conversion of lignocellulosic feedstocks presents significant challenges due to the generation of microbial inhibitors (phenolic compounds, furans, and organic acids) during the hydrolysis pretreatment. These compounds compromise microbial viability and fermentation efficiency, making detoxification a necessary step. This chapter addresses this issue by first reviewing the physical, chemical, and enzymatic pretreatment methods that lead to the formation of inhibitors, characterizing their behavior in the fermentation systems. Then it presents a comprehensive overview of available detoxification strategies, including chemical, biological, and physical methods, following for the potential reactor configurations. Furthermore, the chapter explores sustainability aspects by examining economic and environmental assessments associated with detoxification processes, including life-cycle analysis (LCA) studies that quantify their impact on resources. Finalizing with the technical challenges and new prospects. By integrating these perspectives, the chapter highlights the importance of selecting appropriate detoxification strategies based not only on technical performance but also on their economic and environmental feasibility. It concludes with a discussion of the remaining overall challenges and future perspectives for improving detoxification technologies and their role in advancing sustainable biorefinery processes.