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In this study, the whole plant of Mimosa pudica was extracted using absolute ethanol. The crude ethanolic extract and its isolated triterpenoid glycoside were tested for antifungal activity towards Aspergillus flavus and Trichophyton rubrum using Well Diffusion Method. At concentrations of 25 – 100 mg/mL, the extract possesses antifungal activity (from being partially active to very active) against Aspergillus flavus and Trichophyton rubrum. The TLC profile of the crude extract indicated many compounds with RF values of 0.41, 0.43, 0.56, 0.68, 0.89, and 0.90. The isolation and purification were established using extensive thin layer and column chromatographic processes by employing various solvents with varied polarity. The results of thin layer and column chromatography gave rise to the isolation of a triterpenoid glycoside with RF value of 0.41. The spectral analyses involving 1H-NMR, 13C-NMR, 13C-DEPT, 135COSY, HMBC, HMQC and infrared were useful in proposing the structure of the compound.