The aim of the present investigation was isolation and characterization of active components derived from whole plant of Saccharum spontaneum. The plant was extracted with ethanolic solvent. The preliminary phytochemical results revealed that alkaloids, carbohydrates and glycosides, phenolic compounds, saponins, tannins, protein and amino acids, coumarins & flavonoids as active constituents in ethanolic extract of Saccharum spontaneum. The ethanolic extract of Saccharum spontaneum was undergone column chromatography with different solvent fractions. Despite, three compounds were isolated from ethanolic extract of Saccharum spontaneum with the compound 1 was eluted with pet. ether: benzene 50:50, v/v and compound 2 were eluted with ethyl acetate: ethanol 90:10, v/v and then compound 3 were eluted with ethyl acetate: ethanol, 80:20, v/v. The structures of the three isolated compounds were characterized by using FT-IR, NMR and Mass spectrophotometric methods. Thus, the compound 1 was characterized as 3,3',4',5-Tetra hydroxy-6,8-dimethoxy flavone (C17H14O8), the compound 2 was characterized as 3,5-Dihydroxy-4'-methoxy-7- oxyglucopyronoside flavone (C22H22O11) and the compound 3 was characterized as 3,3',4',5,7- Penta hydroxy flavone (C15H10O7). Furthermore, pharmacological studies required for the isolated compounds.