Ayurvedic formulations have been utilised in healthcare for thousands of years and have made significant contributions to human health protection and improvement. Although its treatments have been used for millennia and are known to be non-toxic or considered non-toxic, there is a lack of verifiable data to support the numerous claims. The current investigation sheds light on the acute and subacute oral toxicity of Dhatupaushtik Churna aqueous extract in Swiss mice and Wistar rats. Acute toxicity assessment was studied using male Swiss mice weighing 20-25 g, and the subacute toxicity testing was done using male Wistar rats weighing
150-200 g. The acute toxicity study was carried out according to OECD (Organisation for Economic Co-operation and Development) guidelines 423. There were no deaths of mice at an oral dose of 2000 mg/kg in the acute toxicity studies. The extract-treated rats (200 and 400 mg/kg, p.o.) showed no significant differences compared to the control group in the subacute toxicity investigation. The liver, kidney, pancreas, and testis weights remained unchanged, while haematological and biochemical markers remained stable. The findings revealed that Dhatupaushtik Churna had no discernible effect on the parameters evaluated at the doses used in the study. However, further investigation is required to confirm its safety.