Qualitative Screening and Analysis of Lunasia amara Blanco (Tawal-ulad) Ethanolic Leaves Extract as Potential Anti-Angiogenic Inhibitors Using the Chorioallantoic Membrane Assay on Mallard Duck Embryo

Angiogenesis, the physiological process of forming new blood vessels, is a hallmark of tumor progression and metastasis in various cancers. As resistance to traditional chemotherapy increases, anti-angiogenic therapies have emerged as promising alternatives. This study investigates the anti-angiogenic potential of Lunasia amara Blanco (Tawal-ulad) ethanolic leaf extract using the chorioallantoic membrane (CAM) assay on mallard duck embryos. A quasi-experimental pretest-posttest design with non-equivalent groups was employed to assess the extracts inhibitory effect on vascular development across five treatment groups (1 mg/mL, 3 mg/mL, 5 mg/mL, pure extract) and two controls (positive: 1 mg/mL Celecoxib; negative: untreated). Results indicated that the pure extract yielded the highest average inhibition (68.89%), particularly in tertiary, quaternary, and quinary blood vessels, suggesting a concentration-dependent inhibitory trend. Despite these observable trends, one-way ANOVA and Tukey's post hoc tests revealed no statistically significant differences (p > 0.01) among treatments, indicating the effects may be due to biological variability. Nonetheless, the pronounced inhibitory patterns in higher extract concentrations support the need for further studies. These findings suggest Lunasia amara holds potential as a natural anti-angiogenic agent pending further validation through advanced assays and larger sample sizes.