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The present study aims to separate the active fractions from the n-hexane extract of Siam weed (Chromolaena odorata (L) R.M. King & H. Rob) leaves with potential as the antiacne agent. The present work includes isolation of active fractions guided by antiacne assay against Propionibacterium acnes and phytochemical analysis of the active fractions. Extraction was conducted by maceration using n-hexane. An agar well diffusion assay was applied to evaluate the antiacne activity of the extract at the concentration of 400 µg/well with chloramphenicol as a positive control. Column chromatography assisted by thin-layer chromatography of n-hexane extract gave six active fractions. Fractions TH5 and TH6 displayed the highest inhibition of 51% and 47% respectively. Further purification of the fraction yielded more than 14 subfractions. Phytochemical analysis indicated that fraction TH5 contains alkaloid and steroid while fraction TH6 contains alkaloid. The active fractions isolated from C. odorata have the potential to be developed as a natural antiacne.