Abstract
<title>Abstract</title> <p> <bold>Introduction:</bold> <italic>Balanites aegyptiaca</italic> is a medicinal plant widely used in traditional medicine, particularly in Africa and southern Asia, for the treatment of infectious diseases and oxidative stress-related conditions, including diarrhea, dysentery, anthrax, wounds, and malignant infections. This study was designed to evaluate the phytochemical composition, antioxidant activity and Antibacterial activity of the aqua-ethanol extract of <italic>Balanites aegyptiaca</italic> leaves using qualitative screening and Liquid Chromatography-Mass Spectrometry/Mass Spectrometry (LC-MS/MS) <bold>Methods:</bold> Qualitative and quantitative phytochemical analyses were conducted using standard procedures. Antioxidant activity was evaluated using the DPPH free radical scavenging assay and the β-carotene-linoleic acid bleaching assay. Antibacterial activity was assessed against <italic>Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa,</italic> and <italic>Klebsiella pneumoniae</italic> using agar well diffusion, minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) assays. LC-MS/MS analysis was performed to identify the major bioactive constituents. <bold>Results:</bold> Phytochemical screening confirmed the presence of alkaloids, flavonoids, tannins, saponins, glycosides, phenols, terpenoids, and carbohydrates, while steroids and proteins were absent. Phenolic compounds were the predominant constituents (42.50 ± 0.01 mg/g extract), followed by flavonoids (35.80 ± 0.02 mg/g), saponins (31.20 ± 0.03 mg/g), and tannins (28.40 ± 0.03 mg/g). The extract exhibited concentration-dependent antioxidant activity, with DPPH radical scavenging increasing from 34.80% to 57.00% and an IC₅₀ value of 33.64 ± 0.04 μg/mL, compared with 19.65 ± 0.02 μg/mL for ascorbic acid. In the β-carotene-linoleic acid assay, antioxidant inhibition increased from 35% to 85%, approaching the activity of BHT (40–92%). The extract also demonstrated concentration-dependent antibacterial activity, with inhibition zones of 16.0–20.0 mm at 100 mg/mL. <italic>Staphylococcus aureus</italic> was the most susceptible organism, whereas <italic>Pseudomonas aeruginosa</italic> was the least susceptible. MIC and MBC values ranged from 20–30 mg/mL and 40–60 mg/mL, respectively. LC-MS/MS analysis identified quercetin, kaempferol, gallic acid, and balanitin-6 as major bioactive constituents. <bold>Conclusion:</bold> The aqueous-ethanolic leaf extract of <italic>Balanites aegyptiaca</italic> is rich in phenolic and flavonoid compounds and exhibits appreciable antioxidant and broad-spectrum antibacterial activities. The identified phytochemicals likely contribute to these biological properties, supporting the potential use of <italic>B. aegyptiaca</italic> leaves as a natural source of antioxidant and antimicrobial agents for pharmaceutical and nutraceutical applications. </p>