Preclinical Safety Assessment of a Polyherbal Formulation: Acute, Subchronic, and Mutagenic Evaluation

Introduction: Before market approval, novel herbal medicines and bioactive compounds require rigorous genotoxicity and safety evaluations. A polyherbal formulation derived from Dasiphora fruticosa, Cynara scolymus, and Rosa acicularis has previously demonstrated antioxidant and nephroprotective properties. However, comprehensive toxicological and mutagenicity evaluations are needed to support its clinical development. In this study, we aimed to evaluate the acute and subchronic toxicities and mutagenic potential of this polyherbal formulation. Methods: Acute oral toxicity was assessed in C57BL/6 mice using a two-phase protocol based on Lorke's method. Subchronic toxicity was evaluated in Wistar rats following OECD guideline 407, with daily oral administration of the polyherbal formulation at doses of 500 and 1,000 mg/kg for 28 days. Mutagenicity was assessed using the Muta-Chromoplate (Ames test) kit according to OECD guideline 471. Results: Acute toxicity evaluation determined that the median lethal dose (LD₅₀) of the polyherbal formulation exceeds 5,000 mg/kg, categorising it as practically nontoxic. The subchronic toxicity assessment revealed that doses of 500 and 1,000 mg/kg had no significant effects on body and organ weight, haematological and biochemical parameters, and histopathological features compared with the controls. Furthermore, the Ames test confirmed that the polyherbal formulation had no mutagenic activity. Conclusions: The polyherbal formulation exhibited no acute toxicity at doses up to 5,000 mg/kg, and no adverse effects were observed in a 28-day subchronic toxicity study. Furthermore, its favourable safety profile was further confirmed by its lack of mutagenic potential. Collectively, these findings provide a robust foundation for continued preclinical and clinical development of the polyherbal formulation