Man ever since his first appearance on earth, has used plant throughout his historical development as a source of medicines. Medicinal plants have formed the basis of the folkloric medicine, which was the main source for new medical discoveries. By the middle of the nineteenth century at least 80% of all medicines were derived from plants. Therefore, in this work, as a result of this intensive study, seventeen phenolic constituents were individually isolated and identified from the methanolic extract of licorice roots by various chromatographic methods (TLC, CC, GC-MS, LC-MS, .. etc). The structure of these compounds was supported by their spectral data and chromatographic analysis as HPLC, GC/MS and PC. On the other hand, the methanolic extract of Licorice roots have been subjected to certain (in vitro) biological studies including the evaluation of the antioxidant activity against the stable free radical DPPH, cytotoxic ctivity on three human cell lines (liver, colon and breast) and its activities as antimicrobial.