Phytochemical profiling, antioxidant evaluation, and anticancer potential of Azolla pinnata methanolic extract: In vitro and in silico insights

Azolla pinnata , a fast-growing aquatic fern, has recently gained attention for its medicinal potential. This study aimed to evaluate the phytochemical composition, antioxidant activity, and anticancer effects of the methanolic extract of A. pinnata . Preliminary phytochemical screening confirmed the presence of flavonoids, phenolics, and saponins, suggesting strong antioxidant potential. Fourier transform infrared (FTIR) spectroscopy (FT-IR) analysis revealed functional groups such as hydroxyl, carbonyl, and amide bonds, while gas chromatography–mass spectrometry analysis revealed diverse phytoconstituents with known pharmacological properties. The antioxidant activity, assessed using the 2,2-diphenyl-1-picrylhydrazyl (DPPH), demonstrated moderate radical scavenging potential with an IC₅₀ value of 124.75μg/mL. Cytotoxicity evaluation using the MTT assay on MCF-7 breast cancer cells showed a dose-dependent reduction in cell viability, with an IC₅₀ of 103.29μg/mL, accompanied by morphological features indicative of apoptosis. Molecular docking analysis of neophytadiene and gamma-sitosterol against apoptotic regulatory proteins (Bax, Bcl-2, p53, and Caspase-3) revealed favorable binding interactions. Notably, γ-sitosterol exhibited stronger binding affinities and diverse interaction profiles, suggesting enhanced pro-apoptotic potential. These findings highlight that A. pinnata as a promising source of bioactive compounds with antioxidant and anticancer properties, warranting further investigation through bioactive isolation, in vivo validation, and mechanistic studies.