Antibacterial substances belong to a group of compounds that attack dangerous microorganisms. Therefore, killing bacteria or reducing their metabolic activity will lessen their adverse effects on a biological system. They originated from either synthetic materials, microbes, or mold. Many of these medications treat the gram-negative bacteria from the critical precedence group, such as Pseudomonas, carbapenem-resistant Acinetobacter, and Enterobacterales. This study aims to investigate the simultaneous analysis of specific antibacterial spectrophotometrically. The WHO maintains this list of priority infections with antibiotic resistance. Drug combinations in single dosage forms are becoming increasingly popular in the pharmaceutical industry. This has created a significant issue for pharmaceutical administrators, particularly in combating fake medications and pharmaceutical analysts, and developing reliable and accurate methodologies with minimal overlapping effects on quantification. The basics of several spectrophotometric methods utilized to conduct multi-component analysis are collected in the current work, and the validation criteria that are an essential component of the approaches are also described. Numerous analytical techniques, including high-performance liquid chromatography, electrochemical methods, flow injection techniques, gas chromatography, spectrofluorometric techniques, capillary electrophoresis, and spectrophotometric techniques, have been reported to determine antibacterials. This study conducts a concise narrative evaluation of the many spectrophotometric methods that have been published for the simultaneous investigation of Levofloxacin, Sulfamethoxazole, Metronidazole, and Rifampicin in their pure forms, pharmaceutical dosage forms, and biological samples due to their sensitivity, simplicity, and cost-effectiveness.