In Vitro and In Vivo Toxicity Profiling of Ammonium-Based Deep Eutectic Solvents

The cytotoxic potential of ammonium-based deep eutectic solvents (DESs) with four hydrogen bond donors, namely glycerine (Gl), ethylene glycol (EG), triethylene glycol (TEG) and urea (U) were investigated. The toxicity of DESs was examined using In Vitro cell lines and In Vivo animal model. IC₅₀ and selectivity index were determined for the DESs, their individual components and their combinations as aqueous solutions for comparison purposes. The cytotoxicity effect of DESs varied depending on cell lines. The IC₅₀ for the Gl_DES, EG_DES, U_DES and TEG_DES followed the sequence of TEG_DES< Gl_DES< EG_DES< U_DES for OKF6, MCF-7, A375, HT29 and H413, respectively. Gl_DES was selective against MCF-7 and A375, EG_DES was selective against MCF-7, PC3, HepG2 and HT29, U_DES was selective against MCF-7, PC3, HepG2 and HT29, and TEG_DES was selective against MCF-7 and A375. However, acute toxicity studies using ICR mice showed that these DESs were relatively toxic in comparison to their individual components. DES did not cause DNA damage, but it could enhance ROS production and induce apoptosis in treated cancer cells as evidenced by marked LDH release. Furthermore, the examined DESs showed less cytotoxicity compared with ionic liquids. To the best of our knowledge, this is the first time that combined In Vitro and In Vivo toxicity profiles of DESs were being demonstrated, raising the toxicity issue of these neoteric mixtures and their potential applicability to be used for therapeutic purposes.