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<p>The range of e-cigarette flavors has driven consumer interest while raising concerns about potential health risks. However, the toxicological impact and chemical composition of flavored e-liquids remain poorly characterized. This study investigates cytotoxicity differences between tobacco- and non-tobacco-flavored e-liquids and identifies key compounds contributing to toxicity through chemical correlation analysis. We assessed the cytotoxicity of 100 e-liquids (50 tobacco-flavored, 50 non-tobacco-flavored) using IC<sub>50</sub> values in NCI-H292 cells and selected representative samples for genotoxicity evaluation. Three component categories—principal constituents, organic acids, and cooling agents—were analyzed, alongside correlations between specific compounds and cytotoxicity. Our findings revealed that non-tobacco-flavored e-liquids exhibited significantly higher cytotoxicity than tobacco-flavored variants, with considerable variation across different flavors and nicotine concentrations. Among non-tobacco flavors, tropical fruit and melon varieties induced the greatest cytotoxicity. Genotoxicity analysis showed that while tobacco-flavored e-liquids (0.2 × to 1 × IC<sub>50</sub>) did not cause significant DNA damage, certain non-tobacco flavors (0.2 × to 1 × IC<sub>50</sub>) triggered pronounced DNA damage. Correlation analysis identified N,2,3-trimethyl-2-isopropylbutamide (WS-23), benzoic acid, and nicotine as negatively correlated with IC<sub>50</sub> values, with WS-23 displaying the strongest association. Further in vitro validation confirmed that WS-23 (0–2.4&#xa0;mg/mL) induces DNA damage and elevates reactive oxygen species (ROS) levels in a dose-dependent manner. These findings highlight the need for more rigorous investigation and regulatory oversight of flavored e-liquids to mitigate potential health risks.</p>

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In vitro toxicological assessment using NCI-H292 cells of the relationship between cytotoxic effects and chemical composition in flavored e-liquids

  • 期刊论文

  • 发表日期：

    2025年5月22日

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  • Shuhong Zhang,
  • Yumeng Zhang,
  • Hongjuan Wang,
  • Shulei Han,
  • Huan Chen,
  • Hongwei Hou

摘要

The range of e-cigarette flavors has driven consumer interest while raising concerns about potential health risks. However, the toxicological impact and chemical composition of flavored e-liquids remain poorly characterized. This study investigates cytotoxicity differences between tobacco- and non-tobacco-flavored e-liquids and identifies key compounds contributing to toxicity through chemical correlation analysis. We assessed the cytotoxicity of 100 e-liquids (50 tobacco-flavored, 50 non-tobacco-flavored) using IC50 values in NCI-H292 cells and selected representative samples for genotoxicity evaluation. Three component categories—principal constituents, organic acids, and cooling agents—were analyzed, alongside correlations between specific compounds and cytotoxicity. Our findings revealed that non-tobacco-flavored e-liquids exhibited significantly higher cytotoxicity than tobacco-flavored variants, with considerable variation across different flavors and nicotine concentrations. Among non-tobacco flavors, tropical fruit and melon varieties induced the greatest cytotoxicity. Genotoxicity analysis showed that while tobacco-flavored e-liquids (0.2 × to 1 × IC50) did not cause significant DNA damage, certain non-tobacco flavors (0.2 × to 1 × IC50) triggered pronounced DNA damage. Correlation analysis identified N,2,3-trimethyl-2-isopropylbutamide (WS-23), benzoic acid, and nicotine as negatively correlated with IC50 values, with WS-23 displaying the strongest association. Further in vitro validation confirmed that WS-23 (0–2.4 mg/mL) induces DNA damage and elevates reactive oxygen species (ROS) levels in a dose-dependent manner. These findings highlight the need for more rigorous investigation and regulatory oversight of flavored e-liquids to mitigate potential health risks.