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Vernonia amygdalina L., commonly utilized in traditional Vietnamese folk medicine for the management of afflictions including liver diseases, hypertension, diabetes, and elevated cholesterol levels. Despite its extensive employment in local herbal remedies, there exists a scarcity of scientific investigations elucidating its pharmacological and therapeutic potential. This research investigated the antioxidant activity of leaf extracts using a range of solvents, including n-hexane, chloroform, ethyl acetate, n-butanol, ethanol, and water. The ethyl acetate extract was identified as the most potent antioxidant, exhibiting EC50 values of 8.42 and 71.88 µg/mL in ferric reduction and DPPH assays, respectively. The hepatoprotective activity, as measured using the cellular lipid peroxidation method of ethanol leaf extract (IC50 = 5.87 µg/mL) is 4.75 times stronger than the trolox. Additionally, the ethanol extract demonstrated significant inhibitory potential against α-amylase and α-glucosidase enzymes (IC50 of 480.00 and 72.53 µg/mL, respectively). The acute oral toxicity study conducted on a mouse model indicates a safety margin at the experimental dose Dmax = 18.44 g extract/kg. Additionally, this study encompassed a leaf microdissection of V. amygdalina, elucidating pivotal morphological features. So, it is safe for further studies and producing functional products for disease prevention. [ABSTRACT FROM AUTHOR]