Nandhi Mezhugu (NM) is a classical Siddha herbo-mineral formulation indicated for all types of Arthritides. Acute toxicity study was performed at 3 dose levels, i.e., 50, 300, and 2000 mg/kg body weight of rats. Sub-acute toxicity study was performed at doses of 9, 45, 90 mg/kg body weight and sub-chronic toxicity study was performed at doses of 9, 45, 110 mg/kg body weight of rats. Palm Jaggery solution was used as vehicle. In sub-acute and sub-chronic toxicity studies, the drug was administered twice for 7 days followed by 7 days of drug holiday, i.e., in sub-acute toxicity study, the period was 56 days and in the sub-chronic toxicity study, it was 180 days. In both studies, the high dose recovery and recovery control groups were used to study the safety of the drug. Biochemical, haematological parameters, gross pathology and histopathology of the rats were analysed in both studies. ICP-OES detection of heavy metal traces in animal tissue samples in sub-chronic toxicity study (high dose group) were analysed. Acute toxicity study showed no mortality and no treatment-related toxicity signs. Similarly, sub-acute and sub-chronic toxicity studies showed no treatment-related abnormalities at the high dose levels such as 90 and 110 mg/kg body weight in rats respectively. Hence, the scientific validation of the safety of Nandhimezhugu is proven by the toxicity studies.