The present study aimed to investigate total flavonoid (TFC), cyanidin-3-glucoside (Cyd-3-glu) content, and anti-oxidant and antibacterial activities of ethanolic seed coat extract of two safflower genotypes (genotype C111 and A82) with contrasting seed coat colors. Despite the absence of Cyd-3-glu in seed coat extracts of white-seeded genotype C111 versus black-seeded genotype A82 and equal TFC index between the two genotypes, there was no significant difference in their antioxidant activity. Also, the ethanolic extract has growth inhibitory properties in pathogenic bacteria. It seems that differences in type and level of secondary metabolites of the seed coat with different color patterns can result in the ethanolic extract’s antioxidant activity. In addition, the results confirmed that seed coat color has not effect on the level (or severity) of the antibacterial properties of ethanolic seed coat extract.