<p>Bacterial biofilms are complex communities in which microorganisms are adhered and encapsulated in a self-produced extracellular polymeric material (EPS) matrix. Biofilm acts as a protective barrier, enhancing the resistance of bacteria to environmental stressors, cleaning agents and antimicrobial treatments. Therefore, this study aimed to evaluated the effectiveness of a mixed disinfectant suspension (MIX) against <i>Staphylococcus aureus</i> biofilms on sandblasted stainless steel disc surfaces. A cubic phase containing MIX was prepared, and its anti-biofilm ability was assessed in comparison to free MIX. The susceptibility of <i>S. aureus</i> ATCC 15,564 (SA^ATCC) and multidrug-resistant <i>S. aureus</i> CCARM 3080 (SA^CCARM) biofilms to single and mixed disinfectants (MIX) was evaluated. The incorporation of MIX into cubosomes was performed to evaluate its potential enhancement in antibiofilm activity. 1% MIX demonstrated the most effective anti-biofilm activity. Extracellular DNA (eDNA) and polysaccharides were also significantly reduced in biofilms treated with 1% MIX. TEM analysis revealed that 1% MIX effectively disrupted bacterial cell clusters within the biofilms. The incorporation of 1% MIX into cubosomes enhanced its antibiofilm activity, with cubosome-encapsulated MIX exhibiting superior results compared to free MIX. This study demonstrates the effectiveness of a mixed disinfectant suspension in reducing <i>S. aureus</i> biofilms on sandblasted stainless steel disc surfaces. Furthermore, the incorporation of MIX into lipid-based nanocarriers, such as cubosomes, enhanced its antibiofilm efficacy compared to the free disinfectant.</p>