Abstract: The aerosol from various market flavored e-liquids and pod-based ENDS flavored formulations (tobacco, menthol and non-tobacco non-menthol) were evaluated for their potential in vitro toxicity.
A closed system pod-based system with temperature-regulated ENDS device and an open tank ENDS (Aegis Mini Mod with a Nautilus Aspire tank and a Nautilus BVC 1.8 Ohms, non-mesh coil) were used to generate the aerosols. All e-liquids contained nicotine-salt at a concentration of 50 mg/mL. The formulations were vaped following CRM81 non-intense puffing conditions and aerosol collected mass (ACM) was extracted using an ethanol extraction methodology. Progressive doses of ACM were tested in the in vitro toxicological assays. The cytotoxicity was assessed by the neutral red uptake in vitro assay in BALBc/3T3 cells (OECD, TG 129). The mutagenicity was assessed by bacteria reverse mutation assay (OECD TG 471) using 5 tester strains of (TA98, TA100, TA102, TA1535, and TA1537) in the presence and absence of rat liver S9 fraction metabolic activation system. The genotoxicity was assessed by MN assay (OECD TG 487) in human lymphoblast TK6 cells.
Under the experimental conditions and based on the established criteria for evaluation of various assays, no aerosol mediated cytotoxicity, mutagenicity or genotoxicity was observed in any of the tested flavors. EC50 for all aerosols could not be calculated for any assay due to the lack of dose-response. The extracted aerosol samples generated either with pod-based temperature- regulated ENDS device or with a open tank ENDS, including three different flavors (tobacco, menthol and non-tobacco-non- menthol) and containing nicotine-salt at 50 mg/mL, did not induce cytotoxic, mutagenic or genotoxic responses.
Manoj Misra, Ed Carmines, Lise Fraissinet
Affiliations: Chemular, Inc, Hudson, MI, USA