This research describes the development of a supercritical fluid extraction (SFE) method to recover aflatoxin B₁ from fortified soil. The effects of temperature, pressure, modifier (identity and percentage), and extraction type were assessed. Using the optimized SFE conditions, the mean recovery from air dried soil was 72%. The variables associated with changes in recovery of aflatoxin were co-solvents, static extraction, and temperature. Acetonitrile–2% acetic acid, used both in-cell and on-line, provided the most efficient recovery. The results indicate that desorption from the soil was the limiting factor in recovery and that the static phase was more important than the dynamic.