Subacute oral exposure to Fumaria parviflora aromatic water (hydrosol) is associated with 2,4-DTBP-related toxicity in rats, with molecular docking suggesting a potential pro-oxidant role

Fumaria parviflora is widely used in traditional Iranian medicine, commonly in the form of hydrosol. Despite its therapeutic applications, data regarding its safety, particularly following repeated exposure, remain limited. This study aimed to evaluate the subacute oral toxicity of F. parviflora hydrosol in a rodent model and to investigate the potential role of its major constituent, 2,4-di-tert-butylphenol (2,4-DTBP), using molecular docking analysis. Hydrosol was prepared via traditional distillation and analyzed by GC–MS to identify its chemical constituents. We administered different doses of hydrosol (0.5, 3.2, and 10 mL/kg) to 40 male and female Wistar rats over a subacute period (28 days). Phytochemical analysis identified seven compounds, with 2,4-DTBP as a major component. High-dose administration (10 mL/kg) resulted in significant histopathological alterations in the liver and kidney, elevated blood urea nitrogen levels ( P < 0.05), and reduced triglycerides ( P < 0.01). White blood cells were significantly ( P < 0.05) decreased, and Mean Corpuscular Hemoglobin was significantly ( P < 0.05) increased in all treated groups. Molecular docking revealed strong binding affinities of 2,4-DTBP toward RXR-α, PPARα, and TNF-α, suggesting a potential pathway to the observed toxic effects. F. parviflora hydrosol exhibits dose-dependent toxicity, with higher doses inducing significant hepatorenal damage. The presence of 2,4-DTBP may contribute to these effects through interactions with key molecular targets. These findings highlight the necessity for dose regulation, safety assessment, and standardization in the use of herbal hydrosols.