Background The two-dimensional MXene, known as titanium carbide (Ti₃C₂), is characterized by its substantial interlayer spacing, extensive surface area, hydrophilic nature, exceptional thermal stability, and outstanding electrical conductivity. These distinctive attributes render Ti₃C₂ an ideal candidate for detecting target analytes and immobilizing biomolecules. Bismuth oxide (Bi₂O₃), an essential compound of bismuth, frequently acts as a foundational element in bismuth chemistry. Its applications are diverse, from fireworks to oxygen gas sensors and solid oxide fuel cells, with particular emphasis on its behaviour under elevated temperatures and pressures. Notably, phase transitions to various polymorphs, which remain metastable at room temperature, have been documented under these conditions, indicating potential for numerous applications. Integrating MXene with Bi₂O₃ composites holds significant promise for advancements in energy-related electronics, sensing technologies, and photocatalytic processes. Objective To optimize the synthesis of titanium carbide-bismuth oxide (Ti₃C₂-Bi₂O₃) nanoparticles to enhance their antimicrobial activity by identifying the best synthesis conditions and assessing their effectiveness against various microbial pathogens. Materials and methods The preparation of Ti₃C₂ MXene involves dissolving lithium fluoride in hydrochloric acid, followed by Ti₃AlC₂ and stirring at 40°C for 48 hours. The resulting pellet is then dispersed in ultrapure water and centrifuged to obtain the MXene dispersion. Bi₂O₃ nanoparticles are prepared by preparing bismuth nitrate pentahydrate in nitric acid and adding sodium hydroxide to adjust the pH. The resulting white precipitate is filtered, washed, and dried before being calcined at 400°C for two hours to produce Bi₂O₃ nanoparticles. The Ti₃C₂-Bi₂O₃ composite is synthesized by adding Bi(NO₃)₃ solution to a 5 mg/mL Ti₃C₂Tx MXene solution. The reaction solution is heated to 160°C, and the resulting black powder is labelled as x% Bi₂O₃/MXene. The antimicrobial efficacy of the nanoparticles is assessed using the disk diffusion method. The zones of inhibition are measured and analyzed as indicators of antimicrobial activity. Results The scanning electron microscopy (SEM) analysis revealed the presence of Bi₂O₃ particles alongside Ti₃C₂ nanosheets, while the X-ray diffraction (XRD) analysis and energy-dispersive X-ray spectroscopy (EDS) confirmed the high crystallinity of the compound. Furthermore, the compound was determined to be impurity-free and demonstrated antimicrobial properties. Conclusion The XRDanalysis confirms the effective integration of various materials and the existence of crystalline phases. SEMprovides insights into the morphology and organization of particles within sheets, whereas EDSassesses the elemental composition and its uniform distribution. These studies demonstrate the synthesis of Ti₃C₂-Bi₂O₃ composites, suggesting their potential for usage in applications involving antimicrobial action.