4-hydroxy-3-(1-(arylimino)ethyl)chromen-2-one were synthesized by condensation of primary aromatic amines with 3-acetyl-4-hydroxy-2H-chromen-2-one. These were characterized by IR, 1HNMR, 13CNMR and mass spectral analysis. In vitro biological screening effects of the investigated compounds were tested against the bacterial species Staphylococcus aureus, Escherichia coli, Salmonella typhi, and Bacillus subtilis by Agar cup method. Fungal species Aspergillus niger, Penicillium chrysogenum, Fusarium moneliforme and Aspergillus flavus by the poison plate method.