Phytochemical Profiling and in vitro Antioxidant, Cytotoxic, and Digestive Enzyme Inhibitory Activities of Ethanolic Fenugreek (Trigonella foenum-graecum L.) Seed Extract

Abstract

Trigonella foenum-graecum L. (fenugreek) is a medicinal and culinary plant valued for its rich phytochemical composition and diverse biological activities. This study investigated the phytochemical profile and in vitro biological activities of an ethanolic fenugreek seed extract using chromatographic and biological assays. GC–MS analysis tentatively identified nineteen volatile constituents, with 2-methyl-2-butenal (36.65%) and p-cymene (17.96%) as the major compounds, while HPLC revealed a phenolic-rich extract. The extract exhibited concentration-dependent antioxidant activity, reaching 85.75 ± 1.73% and 71.79 ± 1.95% scavenging in the DPPH and ABTS assays, respectively, at 1000 µg/mL. MTT evaluation demonstrated preferential cytotoxicity toward HepG2 and Caco-2 cancer cells, with IC₅₀ values of 124.65 and 164.39 µg/mL, respectively, compared with 480.8 µg/mL for normal WI-38 cells. The extract also showed moderate pancreatic lipase inhibitory activity (IC₅₀ = 116.74 µg/mL) compared with orlistat (IC₅₀ = 17.83 µg/mL) and dose-dependently inhibited α-amylase and α-glucosidase. Collectively, these findings demonstrate the extract’s in vitro antioxidant activity, preferential cytotoxicity toward the tested cancer cell lines, and inhibitory effects on pancreatic lipase and carbohydrate-digesting enzymes.