Bioactive substances found in plants, microorganisms and animals have fascinated mankind since time immemorial. This review will focus on the progress that has been made over the centuries and our growing insights. The developments relate to both the discovery and characterization of novel bioactive substances, as well as the ceaseless implementation of refined techniques, the use of high-end instruments and breakthroughs in artificial intelligence with deep learning-based computational methods. As these approaches possess great translational potential, with many applications in different fields, such as therapeutic, diagnostic and agrochemical use, there is a good rationale to continue investing in toxinology-related research.