This paper presents the interaction of benzo[a]pyrene (B[a]P) with Cu(II)-montmorillonite to investigate the formation, evolution and potential toxicity of environmentally persistent free radicals (EPFRs) under dark and visible light irradiation conditions. Degradation of B[a]P and the generated transformative products on clay mineral are monitored by gas chromatography-mass spectrometry (GC-MS) technique. Hydroxyl-B[a]P and B[a]P-diones are observed during the transformation of B[a]P under dark condition. B[a]P-3,6-dione and B[a]P-6,12-dione are the main products under visible light irradiation. B[a]P transformation is accompanied by the formation of EPFRs, which are quantified by electron paramagnetic resonance (EPR) spectroscopy. With increasing reaction time, the concentrations of the produced EPFRs are initially increased and then gradually decrease to an undetectable level. The deconvolution results of EPR spectra reveal formation of three types of organic radicals (carbon-centered radicals, oxygen-centered radicals, and carbon-centered radicals with a conjugated oxygen), which also co-exist. Correspondingly, visible-light irradiation promotes the formation and the decay of these EPFRs. The produced B[a]P-type EPFRs induce the generation of reactive oxygen species (ROS), such as superoxide (O-2(-center dot)) and hydroxide radicals ((OH)-O-center dot), which may cause oxidative stress to cells and tissues of organisms. The toxicity of degradation products is evaluated by the livability of human gastric epithelial GES-1cells. The toxicity is initially increased and then decreases with the elapsed reaction time, which correlates with the evolution of EPFRs concentrations. The present work provides direct evidence that the formation of EPFRs in interaction of PAHs with metal-contaminated clays may result in negative effects to human health.