The assessment of in vitro hemolytic activity of Zornia gibbosa Span involves evaluating its potential to induce the lysis or rupture of red blood cells (erythrocytes) in a laboratory setting. Zornia gibbosa Span, commonly known as "marsh cudweed," is a plant species found in various regions, including tropical and subtropical areas. Research into its hemolytic activity can provide insights into its safety profile and potential pharmacological applications. To assess hemolytic activity, the Zornia gibbosa Span extract is incubated with the red blood cell suspension under controlled conditions, such as temperature and pH. Following the incubation period, the mixture is centrifuged to separate intact red blood cells from any lysed or ruptured cells. The supernatant is then analyzed to measure the release of hemoglobin, a marker of hemolysis, using spectrophotometric methods. The results are expressed as a percentage of hemolysis, calculated based on the absorbance of the supernatant compared to positive and negative controls. Higher absorbance values indicate greater hemolysis, while lower values suggest minimal or no hemolytic activity. Statistical analysis is often performed to assess the significance of the findings and determine dose-response relationships.