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Objective: Silver sulfadiazine and benzalkonium chloride are biocidal agents known to be effective against microorganisms. Although planktonic cells are easily affected by these agents, bacteria in the biofilm are more resistant to the same chemicals. Agents fixed on nanoparticles are reported to have higher antimicrobial activity against biofilm bacteria. The effects of nanotechnological disinfectants on biofilm bacteria vary with dose and time. The type of surface on which these disinfectants are sprayed also changes their activity. The aim of this study was to investigate the ability of silver sulfadiazine and benzalkonium immobilized on nanoparticles to adhere to surfaces and their effect on biofilm. Methods: Stainless steel coupons were sprayed with nano benzalkonium chloride and nano silver sulfadiazine were air-dried to cure for 24 hours, then the coupons were placed into the biofilm reactor. Bacterial counts in the biofilm layer formed on these surfaces were compared at monthly intervals with control coupons. Bacterial biofilms allowed to form naturally were analyzed by microbiological culture methods and number of live and dead microorganisms were determined by DAPI - CTC staining on epifluorescence microscope. Results: After the test periods, microbial culture and DAPI - CTC (live / dead) results were found significantly lower on disinfectant covered coupons. Furthermore, nano benzalkonium chloride was found to be significantly more effective against biofilm bacteria than nano silver sulfadiazine. Conclusion: The results were examined in order to observe biofilm formation on the surfaces covered with nano benzalkonium chloride and nano silver sulfadiazine compounds, to evaluate their use in industrial and clinical fields as effective disinfectants. Because these disinfectants are frequently used in industrial or clinical environments, results have made an important contribution to the manufacturer to focus on final version of the product. [ABSTRACT FROM AUTHOR]