The uses of natural plant origin bioactive compounds are emerging as a promising strategy to detoxify aflatoxin B1 (AFB1). This study aimed to explore the potential of cooking, phytochemicals content, and antioxidant activities derived from garlic, ginger, cardamom, and black cumin to detoxify AFB1 on spice mix red pepper powder (berbere) and sauté. The effectiveness of the samples was analyzed for AFB1 detoxification potential through standard methods for the examination of food and food additives. These major spices showed an AFB1 level below the detection limit. After cooking in hot water for 7 min at 85 ℃, the experimental and commercial spice mix red pepper showed the maximum AFB1 detoxification (62.13% and 65.95%, respectively). Thus, mixing major spices to produce a spice mix red pepper powder had a positive effect on AFB1 detoxification in raw and cooked spice mix red pepper samples. Total phenolic content, total flavonoid content, 2,2-diphenyl-1-picrylhydrazyl, ferric ion reducing antioxidant power, and ferrous ion chelating activity revealed good positive correlation with AFB1 detoxification atp< 0.05. The findings of this study could contribute to mitigation plans of AFB1 in spice-processing enterprises. Further study is required on the mechanism of AFB1 detoxification and safety of the detoxified products.