Effectiveness Test of Pluchea indica L. Leaf Filtrate as a Natural Anticoagulant In Vitro

Abstract

Ethylenediaminetetraacetic acid (EDTA) is a standard anticoagulant used in hematological examinations to prevent blood coagulation and preserve cell morphology. Beluntas leaves (Pluchea indica L.) contain secondary metabolites such as flavonoids, tannins, alkaloids, and saponins that may affect the coagulation process. This study aimed to determine the effectiveness of beluntas leaf filtrate (Pluchea indica L.) as a natural anticoagulant compared with EDTA based on blood clotting time and erythrocyte morphology. This study used a pre-experimental design with a post-test only controlled group design. The filtrate was prepared from fresh and healthy beluntas leaves that were washed, dried, cut into small pieces, ground using a mortar and pestle, then squeezed through a filter cloth and filtered again using filter paper. Venous blood samples were divided into four groups: 500 µL, 600 µL, and 700 µL of beluntas leaf filtrate, and EDTA as the positive control, with six repetitions in each group, resulting in 24 samples. Blood clotting time was observed, while erythrocyte morphology was examined using peripheral blood smears stained with Wright stain at 100× magnification. Data were analyzed using the Kruskal-Wallis test and Fisher's Exact Test. The results showed clot formation on the tube wall at 125, 130, and 140 minutes for 500, 600, and 700 µL of filtrate, respectively, while EDTA showed no clot formation for up to 1440 minutes. Crenation was observed in 83.3% of the 500 µL group and 100% of the 600 and 700 µL groups, whereas all EDTA samples showed normal morphology. Statistical tests showed significant differences in clotting time and erythrocyte morphology (p < 0.001). Beluntas leaf filtrate delayed coagulation but was not yet effective as a substitute for EDTA.