Senna sulfurea is a plant species known for its traditional medicinal uses, including its potential as a laxative. This research aimed to assess in-vitro hemolytic action taken by Senna sulfurea extracts, which is crucial for evaluating its safety and pharmacological effects. Fresh blood samples were obtained combined with RBCs from healthy donors and were separated and incubated with various concentrations of Senna sulfurea extracts. Hemolysis was evaluated by determining the supernatants absorbance to quantify hemoglobin release. The results revealed a concentration- dependent hemolytic effect of Senna sulfurea extracts, with higher concentrations inducing greater hemolysis compared to controls. Furthermore, comparison with positive and negative controls confirmed the hemolytic activity of Senna sulfurea extracts. Mechanistic studies suggested that bioactive compounds present in the extracts, such as saponins and alkaloids, may contribute to RBC lysis. These findings indicate the potential cytotoxic effects of Senna sulfurea on RBCs and raise concerns regarding its safety profile. Additional investigation is necessary to clarify the underlying mechanisms of hemolysis and to assess the safety and efficacy of Senna sulfurea extracts for therapeutic use. Overall, this study provides valuable insights into the pharmacological properties of Senna sulfurea and highlights the importance of considering its hemolytic activity in medicinal applications.