This study investigated the chemical changes of phytochemicals and antioxidant activity during the preparation of traditional Sudanese asida prepared from gamma-radiated (1.0 and 2.0 kGy) flour of two sorghum cultivars (Tabat and Wad Ahmed). For both cultivars, the irradiation process significantly (p < 0.05) increased the total phenolic content and antioxidant activity of the raw flour, while it caused a significant reduction in total flavonoid content and tannin content. Traditional asida (fermented food) prepared from irradiated sorghum flour caused a significant reduction in TPC, TFC, and tannin content in both sorghum cultivars, while the antioxidant activities (DPPH, reducing power, and H₂O₂ scavenging) were significantly increased in both the Tabat (85.0%, 3.8 mg AAE/g, and 84.6%, respectively) and the Wad Ahmed (89.6%, 3.9 mg AAE/g, and 83.1%, respectively) sorghum cultivar grains, particularly in those processed from 2.0 kGy-irradiated flour. A positive high correlation was observed between gamma radiation (2.0 kGy) and the antioxidant activity of asida prepared from both cultivars. In conclusion, traditional fermented asida obtained from irradiated flour showed high antioxidant activity in both sorghum cultivars.