This study purposes to assess the cytotoxicity of 1-dodecyl-3-methylimidazolium chloride ([C 12 min]Cl) in human hepatocellular carcinoma (HepG2) cells. To this end, HepG2 cells were exposed to a range concentration of [C 12 min]Cl and evaluated cell viability, genotoxicity, oxidative stress, apoptosis, cell cycle, and apoptosis-related gene expression to determine cytotoxicity. The outcomes showed that [C 12 min]Cl curbed HepG2 cell growth and reduced cell viability in a concentration-and time-dependent manner.Moreover, our assay results also revealed that exposure to [C 12 min]Cl prompted DNA damage and apoptosis, reduced SOD and GSH content, enhanced MDA level, and changed the cell cycle of HepG2 cells. In addition, [C 12 min] Cl caused alters in the expression levels of p53, Bax, and Bcl-2, indicating that p53 and Bcl-2 family may be involved in the cytotoxicity and apoptosis of HepG2 cells induced by [C 12 min]C1. In summary, these results indicate that [C 12 min]Cl exerts genotoxicity, physiological toxicity and prompts apoptosis in HepG2 cells, and is not an alleged green solvent. K E Y W O R D S apoptosis, cytotoxicity, genotoxicity, ionic liquids, oxidative stress 1 | INTRODUCTION Ionic liquids (ILs) are low melting organic salts at room temperature and are considered a green substitute for industrial volatile organic compounds. 1-3 They are composed of various cations and anions, for example, imidazolium, pyridinium, bromide (Br − ), and chloride (Cl − ). 4 Last several years, ILs have drawn wide attention owing to their unique excellent physical and chemical properties, such as low vapor pressure and nonvolatility, 5,6 good thermal and chemical stability, 7,8and relatively high viscosity. 9 These ILs as environmentally friendly chemicals are famous mainly because their vapor pressure is negligible compared to many inorganic and organic materials. 5,10 Due to their unique properties, ILs are currently being used in a diverse range of applications, including ecology, 11,12 chemical engineering, 13 the medical field, 14 food industry, 15 textiles, 16 and electrochemistry. 17 However, many studies have shown that ILs may be released into aquatic environments via effluent discharges and accidental spills and cause damage to aquatic organisms. 18,19 For example, IL toxicity has been evaluated in microalgae, 20 fish, 21,22 and in Daphnia magna. 23 In addition, several previous studies also showed that ILs are cytotoxic to many classes of cells, such as the human HeLa cell line, 24 and PC12 cells. 25 In contrast, limited information is available on the toxic mechanism of ILs, which is important in risk evaluation.The human hepatocellular carcinoma (HepG2) cell line was derived from a hepatoblastoma at first and retains the structural and functional polarity that hepatocytes exhibit in vivo. 26 It plays a key role in the activation/detoxification of genotoxic procarcinogens and reflects the metabolism of such compounds in vivo better than experimental models with metabolically incompetent cells and exog...