BACKGROUNDS: Phosphorus is a critical nutrient in agriculture, influencing plant growth and nutritional quality.OBJECTIVES: This study, uniquely designed to investigate the effects of phosphorus (P) fertilization levels, sorghum cultivars, and growing locations on phytochemical content and antioxidant activity in sorghum grains, employed four sorghum cultivars (Hakeka, P954063, Tabat, and Tetron) grown under three P levels (0P, 1P, 2P) in two locations (Gezira and White Nile) in Sudan.METHODS: In this study, four sorghum cultivars were grown in two distinct locations in Sudan, employing a split-plot design with three (P) fertilization levels. P was applied as triple super phosphate directly with the seeds, and additional fertilization included urea applied in two split doses. At physiological maturity, representative sorghum panicles were harvested, processed, and analyzed for bioactive compounds and antioxidant activities using standard extraction and quantification techniques such as Folin-Ciocalteu for phenolics and colorimetric flavonoid assays. Antioxidant activities were assessed through various assays, including DPPH and FRAP. Statistical analyses were performed using a three-way ANOVA to examine the effects of cultivar, P level, and location on the measured parameters, supplemented by multivariate analysis to further elucidate the interactions between these factors.RESULTS: Significant interactions (p