This research aims to evaluate the safety of ethanolic extract of Acalypha hispida (A. hispida) leaves with acute toxicity test using 15 female rats strain Sprague-Dawley. A single dose of different doses of extract (2, 4, 8, and 16 g/kg body weight) was administrated orally, and theobservation was conducted for 14 days. The results revealed that the ethanolic extract of A. hispida leaves was relatively harmless (LD50 16 g/kg BW), did not affect body weight, and did not show clinical signs of toxicity during the observation periods. The parameters of blood serumbiochemistry of all extract-treated groups (alanine aminotransferase, aspartate aminotransferase, creatinine, and urea) did not change significantly compared to the control group. The histological observation of the liver showed a significant increase in eosinophilic cytoplasm and basophilic nuclei at all doses. However, the ethanolic extract of A. hispida leaves did not significantly affect glomerulus/Bowman’s capsule ratio, glomerular cell density, and the proportion of normal cell tubule. In conclusion, the ethanolic extract of A. hispida leaves was relatively harmless with LD5016 g/kg BW and seems to be safe in low doses (2 g/kg BW).