Senegalia pennata, a medicinal plant rich in bioactive compounds, has gained attention for its potential therapeutic properties. This study aimed to evaluate the in-vitro heamolytic activity of S. pennata extract. Fresh leaves of S. pennata were collected and extracted using standard procedures. The extract was then subjected to hemolytic activity assessment using erythrocytes obtained from healthy donors. Various concentrations of the extract were incubated with erythrocytes, and the degree of hemolysis was measured spectrophotometrically.Results showed a concentration-dependent hemolytic activity of S. pennata extract. At lower concentrations, minimal hemolysis was observed, while at higher concentrations, a significant increase in hemolytic activity was noted. This suggests a dose-dependent effect of the extract on erythrocytes. Furthermore, the hemolytic activity was found to be influenced by factors such as pH and temperature. this study demonstrates the hemolytic potential of Senegalia pennata extract in vitro. Further investigation into the underlying mechanisms and in vivo studies are warranted to elucidate its therapeutic relevance and safety profile