Putri malu (Mimosa pudica Linn.) is one of plamts that grows in Indonesia. The previous research showed that the extract of Putri malu leaves have an anti-fungal activity. The research aims to determine the anti-fungal activity subfractions of methanol-water fraction of Putri malu using column chromatography againts Aspergillus brasiliensis, Candida albicans, Trichophyton mentagrophytes, and Pityrosporum ovale. Extraction was done with maceration method using methanol:water (70:30) solvent, fractination used n-hexane and ethyl acetate solvents and the methanol-water was further separated with column chromatography. The anti-fungal activity test was conducted using agar diffusion method. The anti-fungal activity of subfractions of methanol-water of Putri malu leaves showed that tose sub-fractions do not have anti-fungal againts A. brasiliensis, C. albicans, T. mentagrophytes, and P. ovale.