Various novel substituted acetyl salicylanilide derivatives were prepared and Characterized all the analytical and spectroscopic data and support the proposed structures. All the seven compounds were scanned against microorganism for their antibacterial activity like Escherichia coil, Pseudomonas putida, Bacillus cereus, Pseudomonas aeruginosa, Staphylococcus aureus and for Antifungal activity of above produce compounds ware scanned against different fungus like Alternative rock, Aspergillus nigar, Rhizoctonia solani. Among them, 2-(4-Sulfamoyl phenylcarbamoyl) phenyl acetate 2-(4(N-(5 methyloxazol, 3-yl) Sulfamoyl) phenyl carbomoyl) phenyl acetate, 2-(4- hydroxyphenol carbomoyl) phenyl acetate gives higher activity in antimicrobial study against E- coil, Staphylococcus Aureus, P. Putida microorganism and 2(4-hydroxyphenol carbomoyl) phenyl acetate, 2-(2-Phenyl hydrazine carbomoyl) phenyl acetate, 2-(4-Sulfamoyl phenylcarbamoyl) phenyl acetate gives higher antifungal activity against Alternaria, R. Solani organism.