Plants and their secondary metabolites are the rich source of drugs against several diseases. Therefore, our aim of study was to isolate and characterized several secondary metabolites from Lawsonia alba (Henna) using state of the art spectroscopic techniques e.g., UV, IR, Mass spectroscopy, 1H- and 13C-NMR spectroscopy and explore the bioactivities of crude plant extract, semipure frcations and pure constituents.