Mushrooms have been used for ages as foods with health benefits and traditional medicines to manage several ailments. While edible mushrooms (EMs) may be picked from the wild or cultivated, the boundary between “edibility” and “nonedibility” is not well defined given the unique array of metabolites found in both edible and nonedible species. As such, mushroom edibility is defined within specific parameters to mitigate adverse impacts. Likewise, since not much is known about the potential emerging food and nonfood industry applications of EMs and their metabolites, research on EMs has increased in the past decade to cover the knowledge gap. In the same vein, research giving deeper insights on EMs metabolomic profiles, biological activity range, metabolite biosynthetic pathways, and mechanisms of actions have evolved. This is closely linked to the continued fine-tuning of metabolomic profiling workflows, databases and platforms for analyzing metabolomic data, and increasing precision and accuracy in metabolite identification and annotation. Given these highlights, this chapter focuses on providing a comprehensive insight on metabolomic workflow types and their use in EM metabolite profiling, the challenges associated with them, and an outlook on current, emerging, and/or future applications of metabolomic platforms, EM extracts, and metabolites in various industries. In addition, the collation highlights study gaps in EM metabolomics to guide future research, while touching on metabolites defining the border between edible and inedible mushroom with a view to giving a holistic overview on the topic under investigation.