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<p>The increasing incorporation of multiple sweeteners into processed foods underscores the need to evaluate their combined genotoxicity. Epidemiological studies associate high intake of sweeteners with elevated risks of cardiovascular, cerebrovascular, and metabolic disorders, yet the genetic consequences of their combined consumption remain largely unexplored, highlighting the necessity of assessing their interactive genotoxic risks. This study therefore aimed to investigate the genotoxic and developmental toxicity of the widely used artificial sweeteners, acesulfame potassium (ACE K), aspartame (ASP), and stevia (S), individually and in relevant binary and ternary combinations (ACE K + ASP, ACE K + S, ASP + S, and ACE K + ASP + S) at 10, 30, and 50&#xa0;mM. Genotoxicity was assessed using the Drosophila SMART assay, complemented by molecular docking and 100-ns dynamic simulations with the developmental regulators Engrailed (En) and Hedgehog (Hh), and toxicity prediction via ProTox-3.0. All sweeteners produced dose-dependent genotoxic effects, with ACE K–containing mixtures showing higher spot frequencies at 10&#xa0;mM, while ASP + S exhibited stronger genotoxicity at 30 and 50&#xa0;mM; the triple combination consistently mirrored the 10&#xa0;mM trend. In silico analyses revealed higher binding affinities of ACE K toward En and Hh, aligning with in vivo observations. Collectively, these findings demonstrate concentration-dependent genotoxic risks associated with sweetener mixtures and indicate that formulations incorporating stevia may offer relatively safer alternatives for food applications.</p>

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Genotoxicity of Artificial Sweetener Combinations: An Integrated Drosophila melanogaster SMART Assay and Computational Modeling Approach

  • 期刊论文

  • 发表日期：

    2026年3月17日

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  • Nivedaa Shankar,
  • B. Priyanka,
  • D. Vidhya Janani,
  • Priyanka Balaji,
  • K. Nivetha,
  • J. Vijayalakshmi,
  • B. Amshumala,
  • G. Tamizh Selvan,
  • Solomon F. D. Paul

摘要

The increasing incorporation of multiple sweeteners into processed foods underscores the need to evaluate their combined genotoxicity. Epidemiological studies associate high intake of sweeteners with elevated risks of cardiovascular, cerebrovascular, and metabolic disorders, yet the genetic consequences of their combined consumption remain largely unexplored, highlighting the necessity of assessing their interactive genotoxic risks. This study therefore aimed to investigate the genotoxic and developmental toxicity of the widely used artificial sweeteners, acesulfame potassium (ACE K), aspartame (ASP), and stevia (S), individually and in relevant binary and ternary combinations (ACE K + ASP, ACE K + S, ASP + S, and ACE K + ASP + S) at 10, 30, and 50 mM. Genotoxicity was assessed using the Drosophila SMART assay, complemented by molecular docking and 100-ns dynamic simulations with the developmental regulators Engrailed (En) and Hedgehog (Hh), and toxicity prediction via ProTox-3.0. All sweeteners produced dose-dependent genotoxic effects, with ACE K–containing mixtures showing higher spot frequencies at 10 mM, while ASP + S exhibited stronger genotoxicity at 30 and 50 mM; the triple combination consistently mirrored the 10 mM trend. In silico analyses revealed higher binding affinities of ACE K toward En and Hh, aligning with in vivo observations. Collectively, these findings demonstrate concentration-dependent genotoxic risks associated with sweetener mixtures and indicate that formulations incorporating stevia may offer relatively safer alternatives for food applications.