Fractionation, Phytochemical Profiling, and Antioxidant-Antimicrobial Activities of

Tadehagi triquetrum L., locally known as Duduk leaves, has long been used in traditional medicine for various treatments, including treating fever, liver disorders, and kidney ailments. However, systematic phytochemical and bioactivity studies on this species remain scarce. Unlike previous reports that mainly focus on crude extracts, this study integrates polarity-based fractionation, quantitative phytochemical analysis, antioxidant evaluation, antimicrobial testing, and Liquid Chromatography– Mass Spectrometry (LC-MS) profiling to provide a more comprehensive assessment of the bioactive potential of T. triquetrum L. leaves. The crude extract obtained from maceration with 70% ethanol yielded 28.49%. Subsequent fractionation using n-hexane, chloroform, ethyl acetate, and water produced yields of 6.52%, 0.90%, 12.82%, and 52.37%, respectively. Phytochemical screening revealed that the ethyl acetate fraction contained the highest levels of phenolics, tannins, and flavonoids. This fraction exhibited the strongest antioxidant activity in the DPPH assay (IC₅₀ = 0.71 μg/mL). LC–MS profiling identified 15 putative compounds, with catechin (26.84%) and kaempferol (14.58%) as the dominant constituents. The ethyl acetate fraction also demonstrated moderate to strong antibacterial activity against Staphylococcus aureus and Escherichia coli. These findings highlight T. triquetrum L. as a promising source of natural antioxidant and antimicrobial agents.