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This work aimed to determine and identify the total phenolic (TP) and flavonoids (TF) compounds in different Teucrium polium leave extracts and evaluation their antioxidant and cytotoxic activity against MCF-7 breast cancer cells. Polyphenols were determined by HPLC; however, the TF and TP were estimated by colorimetric methods using rutin and gallic acid as standards, respectively. The HPLC analysis of the methanolic, ethanolic and aqueous extracts identified a total of 9, 8 and 11 compounds, respectively. Naringenin, quercetin, and ellagic acid were the most abundant, and chlorogenic acid, pyrocatechol. Chlorogenic acid, pyrocatechol and 4`.7-Dihydroxyiso flavone was completely absent in the methanolic and ethanolic extracts; however, catechin was completely absent in the ethanolic and aqueous extracts. The methanolic extract exhibited high antioxidant activity then the ethanolic and aqueous extracts. The three extracts exhibited strong cytotoxic activity against MCF-7 cell line using MTT in a dose-dependent fashion and the methanolic extract was more effective with an IC50 of 14.20 ± 1.3 μg/ml. The obtained data indicated that T polium could be an ideal alternative to be used for the development of new anticancer agents due to its potential antioxidant and cytotoxic capacities.