ObjectiveTo isolate and identify the chemical constituents of the EtOAc fraction of Tithonia diversifolia.MethodsThe dried aerial parts of Tithonia diversifolia were extracted with 80% EtOH,and the chemical constituents of the ethyl acetate portion from the extract were isolated by column chromatography with silica gel, Sephadex LH-20 and MCI.The isolated compounds were identified by NMR and MS analysis.ResultsTen compounds were isolated from EtOAc extract, and identified as:vanillin(1), ergosterol(2), ergosterol peroxide(3), 3-methoxy-4-hydroxy-trans-cinnamaldehyde(4), p-hydroxybenzaldehyde(5), 7-methylesculetin(6), 5-hydyoxymethyl furfural(7), nepetin(8), docosanoic acid 2, 3-dihydroxypropyl ester(9) and 13-hydroxyeupatorochromene(10).ConclusionVanillin, ergosterol, ergosterol peroxide, 3-methoxy-4-hydroxy-trans-cinnamaldehyde, p-hydroxybenzaldehyde, 7-methylesculetin, 5-hydyoxymethyl furfural, docosanoic acid 2, 3-dihydroxypropyl ester, and 13-hydroxyeupatorochromene have been isolated from Tithonia diversifolia for the first time.