The chemical composition of Anacyclus valentinus essential oil from Algeria was determined and its antifungal effect in vitro toward 7 toxigenic fungal strains was evaluated. Essential oil yield of 0.13% (v/w) was obtained from the aerial parts of A. valentinus using hydrodistillation. The GC/MS analysis identified 29 components of which delta-3-carene (31%), spathulenol (14.2%), decanoic acid (5.5%), delta-cadinene (4.4%), anethole (3.4%) and aromadendrene (3.3%) were the major compounds of this oil. The results of the antifungal activity showed that the most sensitive fungal strains to A. valentinus oil were Fusarium graminearum, Aspergillus parasiticus, and Penicillium expansum with lower minimal fungicidal concentration (MFC) of 1.25 mu l/ml (v/v). The oil also had a strong fungicidal effect against A. haves and F. moniliforme at a MFC value of 2.5 mu l/ml, while the oil concentration of 10 mu l/ml was needed to show a fungicidal activity against A. ochraceus and P. citrinum. This study suggested that A. valentinus oil is a potential candidate to be used as a safe biocontrol agent to prevent food crops from fungal diseases and improve product quality and safety.