*Article* **Fabrication of Caseinate Stabilized Thymol Nanosuspensions via the pH-Driven Method: Enhancement in Water Solubility of Thymol**

**Wei Zhou 1,2, Yun Zhang 1, Ruyi Li 2, Shengfeng Peng 1,3,\*, Roger Ruan 1, Jihua Li 2 and Wei Liu 1**


**Abstract:** Thymol has been applied as a spice and antibacterial agen<sup>t</sup> in commercial products. However, the utilization of thymol in the food and pharmaceutical field has recently been limited by its poor water solubility and stability. In this work, a caseinate-stabilized thymol nanosuspension was fabricated by pH-driven methods to overcome those limitations. Firstly, the chemical stability of thymol at different pH value conditions was investigated. The physiochemical properties of thymol nanosuspensions were then characterized, such as average particle size, zeta potential, encapsulation efficiency, and loading capacity. Meanwhile, the X-ray diffraction results showed that thymol was present as an amorphous state in the nanosuspensions. The thermal stability of thymol was slightly enhanced by encapsulation through this process, and the thymol nanosuspensions were stable during the long-term storage, and the average particle size of nanosuspensions showed that there was no aggregation of nanosuspensions during storage and high temperature. Finally, the antimicrobial activity of thymol nanosuspensions was evaluated by investigating the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against *Salmomella enterca*, *Staphlococcus aureus*, *Escherichia coli*, and *Listeria monocytogenes*. These results could provide useful information and implications for promoting the application of thymol in food, cosmetic, and pharmaceutical commercial products.

**Keywords:** thymol; nanosuspensions; caseinate; pH-driven method; water solubility
