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The most widespread Sanguisorba species are Sanguisorba officinalis L. and Sanguisorba minor Scop. which are also found in the Romanian flora and classified as medicinal plants because of hemostatic, antibacterial, antitumor, antioxidant and antiviral activities. This study aimed to characterize and compare Sanguisorba species in order to highlight which species is more valuable according to phenolic profile and antimicrobial activity. Based on high-performance liquid chromatography equipped with photodiode array detection and mass spectrometry (electrospray ionization) (HPLC–DAD-MS (ESI+)) analysis, it was evident that the ethanol extract obtained from the leaves of S. minor Scop. contains the highest content of phenolic compounds at 160.96 mg/g p.s., followed by the flower and root extract (131.56 mg/g dw and 121.36 mg/g dw, respectively). While in S. officinalis, the highest amount of phenols was recorded in the root extract (127.06 mg/g), followed by the flower and leaves extract (102.31 mg/g and 81.09 mg/g dw, respectively). Our results show that among the two species, S. minor Scop. is richer in phenolic compounds compared with the S. officinalis L. sample. In addition, the antimicrobial potential of each plant organ of Sanguisorba species was investigated. The ethanol extract of S. minor Scop. leaves exhibited better antibacterial activity against all of the bacteria tested, especially on Staphylococcus aureus, with an inhibition zone of 15.33 ± 0.83 mm. Due to the chemical composition and antimicrobial effect, the Sanguisorba species can be used as food supplements with beneficial effects on human health. [ABSTRACT FROM AUTHOR]