Safety assessment of AYUSH-64 through acute and 90-days repeated dose oral toxicity

Background: AYUSH-64 is a polyherbal Ayurvedic antimalarial formulation, also found effective in COVID-19. Aim: The present study was designed to investigate the safety of AYUSH-64 in acute and 90-day repeated dose oral toxicity as per the Organization for Economic Cooperation and Development (OECD) guidelines 423 and 408. Materials and Methods: During the acute toxicity study, AYUSH-64 was orally administered at a single limit dose of 2000 mg/kg with distilled water as a vehicle. The mortality and clinical signs were observed for 14 days. Similarly, the 90-day repeated dose toxicity was studied by administering the AYUSH-64 at doses of 500, 1000, and 1500 mg/kg. Results: In the acute toxicity study, there were no signs of toxicity. During the 90 days of repeated dose toxicity (subchronic toxicity study), AYUSH-64 at the maximum dose level of 1500 mg/kg did not show any signs of toxicity compared to the control. Histopathological or hematological and biochemical parameters of AYUSH-64 at 1500 mg/kg did not show any significant difference compared to the control (p > 0.05). The organ weights of the test group do not differ significantly compared to the control group. Conclusions: It was concluded that the No Observable Adverse Effect Level (NOAEL) of AYUSH-64 was calculated as 1500 mg/kg/day.