Unveiling the Genotoxic Safety and Antioxidant-Mediated DNA Protective Mechanisms of Triammonium Citrate: An Integrated In Vitro and In Silico Approach

Triammonium citrate (TAC) is a citrate salt widely utilized in diverse industrial and dietary applications, leading to widespread human exposure. While related citrate salts have established safety profiles, the specific genotoxic potential and molecular interactions of TAC remain largely unexplored. This study provides the first integrated in vitro and in silico mechanistic safety assessment of TAC. Cell viability, evaluated in NIH/3T3 and MCF-7 cells via the CCK-8 assay, remained above 90% at all tested concentrations (0.25-2 mg/mL). Furthermore, a comprehensive genotoxicity battery-including the Ames test, chromosomal aberration, micronucleus, and sister chromatid exchange assays in human lymphocytes-revealed no mutagenic, clastogenic, or aneugenic activity. Beyond its safety profile, TAC exhibited marked chemoprotective activity, preserving supercoiled plasmid DNA with up to 96% protection against H₂O₂-induced oxidative damage. Molecular docking studies substantiated these findings by predicting moderate binding affinities for DNA replication enzymes (DNA polymerase δ: ΔG = -5.7 kcal/mol; topoisomerase I: ΔG = -5.9 kcal/mol) while showing a preferential affinity for the metabolic enzyme glutamine synthetase (ΔG = -6.1 kcal/mol). These in silico results suggest that TAC does not interfere with critical DNA maintenance machinery. Collectively, these findings establish a robust safety profile for TAC and unveil a previously unrecognized, antioxidant-mediated DNA-protective function, highlighting its potential as a bioactive functional ingredient capable of mitigating oxidative DNA stress.