Given the heightened awareness of the need to protect the environment, the scientific community has turned to develop green technologies that will have a less harmful effect on the environment. Eutectic solvents stand out as one of the new green solvents, and if such a solution is prepared with the active substance as one of the components, then it is a therapeutic eutectic solvent (THEDES, Therapeutic Deep Eutectic Solvents). Therefore, the aim of this study was to examine the antimicrobial effect of the components that make up THEDES and the antimicrobial effect of THEDES itself on some of the most common representatives of human pathogens: Escherichia coli 3014, Staphylococcus aureus 3048, Pseudomonas aeruginosa 3024, and Candida albicans 86. This test was performed by the disk-diffusion method. The obtained results indicate the possibility of a synergistic antimicrobial effect of octanoic acid and thymol according to E. coli 3014, since no inhibition of bacterial growth was observed in the presence of these components, while in the presence of a mixture of octanoic acid and thymol antibacterial activity is observed. The inhibitory effect of THEDES on all test microorganisms was observed, and E. coli 3014 proved to be the most sensitive to THEDES.