Mycotoxins are naturally occurring toxins produced by certain fungi. Exposure to mycotoxins may occur through the consumption of contaminated foods or from animals that are fed contaminated feed. To safeguard the nation’s food supply, the U.S. Food and Drug Administration (FDA) regulates and monitors mycotoxins in food and conducts research on how best to detect and prevent mycotoxin contamination. Mycotoxin analysis is at the center of the mycotoxin program – concentration data are needed for determining the occurrence of regulated and toxic mycotoxins in food and feed products, estimating dietary exposure, the establishment of guidelines for non-regulated mycotoxins, and accumulating historical data to explore the distribution of mycotoxins as a function of agricultural and food processing practices, geographic location, and environmental conditions. This presentation will discuss the FDA’s continuous efforts to develop and incorporate fit-for-purpose analytical tools for mycotoxin analysis with a focus on the relationship between analytical methodologies and scientific assessments. The presentation will further highlight the Agency’s proactive approaches to address challenges in sampling and analytical method development. Lastly, as environmental factors (e.g., weather, humidity, climate change) and anthropogenic activities (e.g., agricultural and food processing practices) continue to change, affecting the distribution of mycotoxins in foods, the presentation will explore future possibilities to develop new analytical tools to meet emerging regulatory needs.