Globularia alypum leaves have been widely used for more years in gastrointestinal disorders as a traditional folk medicine. The aim of the present study was to determine the chemical composition of the petroleum ether extract and to evaluate its antioxidant activities in comparison with eugenol and its derivative isoeugenol. After phytochemical tests, a simple hydrodistillation was effectuated by Clevenger apparatus and the distillate was extracted with petroleum ether by decantation process. Gas chromatography mass spectroscopy was used to identify and quantify phenolic compounds in this extract. The antioxidant activity of petroleum ether extract of Globularia alypum was measured in vitro by 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging method. However, total antioxidant capacity (Molybdate phosphate test), hydrogen peroxide scavenging and reducing power antioxidant were estimated. The petroleum ether extract demonstrated a low free radical scavenging capacity compared to eugenol and isoeugenol extracted from Syzygium aromaticum. Furthermore, the analysis of this extract by CPG/ms led to the identification of new known phenol named eugenol and also its derivative isoeugenol with considerable amounts (10.56%, 0.87%), respectively. The antioxidant capacity of the petroleum ether extract is probably associated with phenolic compounds detected and its principle compounds indicate that this plant may be an important source of chemopreventive and chemotherapeutic natural products. The best of our knowledge is the combination of new detected compounds for the first time, eugenol and its derivative in this plant which have been tested separately as powerful antioxidant agents. However, further studies are required to determine if this is of clinical significance.