A series of homoleptic and heteroleptic bismuth(III) flavonolate complexes derived from six flavonols of varying substitution have been synthesised and structurally characterised. The complexes were evaluated for antibacterial activity towards several problematic Gram‐positive (Staphylococcus aureus, methicillin‐resistant Staphylococcus aureus (MRSA), and vancomycin‐resistant Enterococcus (VRE)) and Gram‐negative (Escherichia coli, Pseudomonas aeruginosa) bacteria. The cell viability of COS‐7 (monkey kidney) cells treated with the bismuth flavonolates was also studied to determine the effect of the complexes on mammalian cells. The heteroleptic complexes [BiPh(L)2] (in which L=flavonolate) showed good antibacterial activity towards all of the bacteria but reduced COS‐7 cell viability in a concentration‐dependent manner. The homoleptic complexes [Bi(L)3] exhibited activity towards the Gram‐positive bacteria and showed low toxicity towards the mammalian cell line. Bismuth uptake studies in VRE and COS‐7 cells treated with the bismuth flavonolate complexes indicated that Bi accumulation is influenced by both the substitution of the flavonolate ligands and the degree of substitution at the bismuth centre.