Antioxidants are known to play a crucial role in maintaining cellular health by neutralizing harmful free radicals. Among various dietary sources of antioxidants, wheat-based products, particularly flours, have gained significant attention due to their potential health benefits. Durum wheat, a widely cultivated species, serves as a primary ingredient in numerous food products. However, limited research has been conducted to assess the antioxidant activity of flours obtained from durum wheat varieties. In this study, we aimed to comparatively evaluate the antioxidant and antiplatelet potential of flours from 22 selected durum wheat varieties cultivated in Greece. We focused on three major parameters for antioxidant activity measurement: total phenolic content, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, and ferric reducing antioxidant power (FRAP). The total phenolic content was determined using the Folin-Ciocalteu method, while the DPPH and FRAP assays were employed to assess the ability of flours to scavenge free radicals and reduce ferric ions, respectively. Antiplatelet activity was evaluated by the platelet activating factor inhibition assay (PAF) in platelet rich plasma. Analysis of the data revealed notable differences in total phenolics, antioxidant and antiplatelet activities among the tested samples. The total phenolic content ranged from 624.0 + 3.5 to 950.0 + 5.3 μg of gallic acid equivalent/g flour with the variety Zeta E having the highest content. Antioxidant activities based on the DPPH and FRAP assays ranged from 0.56 + 0.02 to 2.26 + 0.08 and 1.93 + 0.02 to 3.65 + 0.03 μmol of trolox equivalent/g flour respectively with the varieties Marco Aurelio and Zeta E exerting the highest antioxidant activities in DPPH and FRAP, respectively. In addition, the IC50 values for antiplatelet activity ranged from 0.72 + 0.21 to 3.06 + 0.17 in mg of flour, with the variety of Zoi exhibiting the highest antiplatelet activity. Overall, this comparative evaluation highlights the differences of antioxidant and antiplatelet activities among flours obtained from 22 different durum wheat varieties cultivated in Greece. The results from this study aid in the selection of wheat varieties with superior antioxidant and antiplatelet capacities for use in bakery food formulation and dietary recommendations.