The aim of the present study was to isolate and characterize the active components present in the root of Lawsonia inermis. The plant was extracted with ethanolic extract. The phytochemical results reveals the presence of carbohydrates, glycosides, tannins, phenolic compounds, flavonoids and terpenes in the plant. The ethanol extract of Lawsonia inermis was undergone coloumn chromatography with different solvent fractions. Two compounds were isolated from ethanolic extract. Compound 1 was eluted with benzene: Chloroform 10:90 v/v and compound 2 was eluted with ethyl acetate:ethanol, 30:70, v/v. The structures of the 2 compounds was characterised by using FT-IR, NMR and Mass spectrometry. Compound 1 was characterised as 24β-ethyl cholest-4-en-3β-ol as C29H50O and compound 2 is 3,7,4’,5'-Tetrahydroxy-6-methoxyflavone as C16H12O7.Therefore, further biological investigation need to carry out isolated compounds present in this plant.