We found that the presence of either vanillin or cinnamaldehyde in e-liquids was associated with higher toxicity values. Our data indicated that e-liquids are extremely heterogeneous, so we also performed gas chromatography–mass spectrometry (GC-MS) of all e-liquids to evaluate their composition/toxicity relationship. In this work, we developed a high-throughput screening (HTS) assay to rapidly triage and validate the toxicity of multiple e-liquids in parallel. There are currently over 7,700 different flavored e-liquids that are commercially available, but there is very limited information regarding either their chemical composition or toxicity. The e-liquids used in electronic cigarettes (E-cigs) typically consist of a mixture of propylene glycol (PG), vegetable glycerin (VG), and nicotine, as well as numerous chemical additives that are used for flavoring. Such an approach may serve as a roadmap to enable bodies such as the Food and Drug Administration (FDA) to better regulate e-liquid composition. Our data suggest that an HTS approach to evaluate the toxicity of multiple e-liquids is feasible. We have also developed a publicly available searchable website ( Given the large numbers of available e-liquids, this website will serve as a resource to facilitate dissemination of this information. Further analysis of common constituents by electron ionization revealed that the concentration of cinnamaldehyde and vanillin, but not triacetin, correlated with toxicity. Furthermore, these data indicated that (i) the more chemicals contained in an e-liquid, the more toxic it was likely to be and (ii) the presence of vanillin was associated with higher toxicity values. Subsequent nonmetric multidimensional scaling (NMDS) analysis revealed that e-liquids are an extremely heterogeneous group. We also performed gas chromatography–mass spectrometry (GC-MS) analysis on all tested e-liquids. Our data demonstrated that the PG/VG vehicle adversely affected cell viability and that a large number of e-liquids were more toxic than PG/VG. Here, we developed a 3-phase, 384-well, plate-based, high-throughput screening (HTS) assay to rapidly triage and validate the toxicity of multiple e-liquids. There are currently over 7,700 e-liquid flavors available, and while some have been tested for toxicity in the laboratory, most have not. The e-liquids used in electronic cigarettes (E-cigs) consist of propylene glycol (PG), vegetable glycerin (VG), nicotine, and chemical additives for flavoring.
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