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Pulp Out, paste-contained jatropha sap, sidaguri roots, and melittin, has been studied to have potency used as an herbal-based devitalizing agent. Prior to clinical application, the toxicity of Pulp Out should be evaluated as it might leak from the cavity unintentionally and get into the digestive systems, which can cause either local or systemic effects. The present study aimed to evaluate the impact of Pulp Out application on periodontal and periapical tissue as well as acute toxicity in Wistar rats. The paste was inserted into the periapical tissue. After 7 days, the periapical tissue was isolated for histopathological evaluation. Pulp Out in an oral suspension of 50, 500 and 2500 mg/kg BW was administered. Autonomic nerve signs were observed intensively every 4 h as well as water and food consumption for 14 days. Biochemical, hematologic parameters and specific organs were evaluated. Therefore, considering the inflammatory lymphocyte cells, osteoblasts, and osteoclasts, Pulp Out is not toxic. The acute toxicity study revealed no treatment-related death was observed, indicating that LD 50 is greater than 2500 mg/kg BW. No significant difference statistically either in body weight, water or food consumption as was observed in autonomic clinical signs of treated groups when compared to the control. Similarly, biochemical and hematologic properties showed no significant difference compared to control. Histopathological, slightly hydrophilic degenerative was observed in all organs. In conclusion, Pulp Out showed low acute toxicity in Wistar rats., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Maria Tanumihardja has patent #P00202007885 issued to Ministry of Law and Human Rights., (© 2022 The Authors.)