This in vitro study was conducted to evaluate the impact of gamma-irradiation on the antioxidant activity of the methanolic extracts of Crocus sativus petals. After gamma-irradiation at 5, 10, and 35 kGy doses, Folin-Ciocalteu, aluminum chloride colorimetric methods, Fourier transform infrared spectroscopy (FTIR), and high performance liquid chromatography (HPLC) techniques were deployed to analyze the irradiated and non-irradiated extracts. The obtained results were indicative of boosted levels of phenolic and total flavonoids compounds in methanolic extracts irradiated at 5 and 10 kGy. Results of the HPLC method also revealed that gamma-irradiation, especially at 5 kGy, could increase the quantity of certain polyphenols, including chlorogenic acid, epicatechin caffeic acid, and thioxanthene, in the saffron extract. In contrast, gamma-irradiation at 35 kGy exerted certain adverse effects on polyphenols contents. In conclusion, gamma-irradiation, especially at 5 kGy, could enhance the antioxidant potential of the methanolic extract of saffron petals.