Biofilm development causes immense economic and social problems for many industries, including the food industry. Biofilm formation of foodborne pathogens in food processing environments can contribute to the persistence and aid in transmission of food-borne diseases. Salmonella are a major cause of food-borne illnesses throughout the world. Salmonella gastroenteritis is usually a self-limiting disease; therefore, antibiotics are not commonly used and so effective sanitization plays a crucial role in preventing contamination and infection. Chemical disinfectants remain the most common mode of microbial inactivation due to costs and practicality reasons. Whilst antibiotic resistance in biofilms is of an area of intensive research, there is less focus on chemical disinfectant resistance in biofilms. Here, we successfully used the MBEC™ Assay to grow and test Salmonella Typhimurium biofilms against disinfectants including benzalkonium chloride, chlorhexidine gluconate, citric acid, quaternary ammonium compounds, sodium hypochlorite and ethanol. Our results showed that all disinfectants reduced biofilm cells at concentrations and contact times sufficient to eliminate planktonic cells, but there were still sufficient viable cells present to cause further contamination and potential infection. Surprisingly, we also demonstrated that a low concentration of sodium hypochlorite was more effective against the biofilm compared to higher concentrations of sodium hypochlorite. The susceptibility of biofilms at different ages to disinfectants was also evaluated and it was concluded that under the conditions of this study, age of biofilm does not contribute to increased resistance towards disinfectants. Rather, the concentration and contact time were both shown to play a role in successful sanitization. Control of Salmonella in food processing environments poses a major challenge to both consumers and producers. The extensive use of chemical disinfectants in the role of sanitization pinpoints the importance in assessing the efficacy of biocides and disinfectants against biofilms of foodborne pathogens. Finally, it is crucial that we adopt a standardised assay designed for high throughput growth and testing of biofilms against disinfectant and biocides.