The present study was carried out to characterize the bioactive constituents present in seed and whole plant extracts of Lepidium sativum using UV-VIS and GC-MS. The crude extracts were scanned in the wavelength ranging from 200 to 800 nm by using Perkin Elmer spectrophotometer and the characteristic peaks were detected. For GC-MS analysis about 25g of powdered plant material was uniformly packed into a thimble and extracted with 150ml of ethanol as solvent using this plant extract was prepared. Helium gas (99.999%) was used as the carrier gas at constant flow rate 1ml/min and an injection volume of 2?l was employed (split ratio of 10:1); Injector temperature 80°C; Ion-source temperature 250°C. The oven temperature was programmed from 110°C (isothermal for 2 min.), with an increase of 10°C/min, to 200°C, then 5°C/min to 250°C, ending with a 9min isothermal at 280°C. Mass spectra were taken at 70 eV; a scan interval of 0.5seconds and fragments from 45 to 450 Da. The UV-VIS profile showed different peaks ranging from 280 and 290 nm with absorbance values of 0.26 and 3.98 respectively. The spectra for phenolic compounds (tannins) and flavonoids typically lie in the range of 230-290 nm. The results of the GC-MS analysis provide different peaks determining the presence of 28 phytochemical compounds in seed extract and the major phyto constituents were (Peak area 16.23%),o-ethyl S-2-Dimethylaminoethyl Ethylphos, (14.37%) Oleoyl chloride and (12.50%) cis-9-Hexadecenal (8.97%). Phytochemical compounds present in whole plant extract was 79 and the major phyto constituents were Eugenol (7.69 % ); Hexadecanoic Acid, Ethyl Ester (7.50%) and Stigmast-5-EN-3-OL, (3.BETA.)- (7.14 %) were reported by GC-MS analysis. The results revealed the major compounds are fatty acid esters and alkaloids which showed antioxidant, antimicrobial and anticancer activities.