Agrimonia eupatoria L. (Rosaceae fam.) is widespread in temperate regions. In folk medicine, this species has been used for its astringent, analgesic and anti-inflammatory, as well as in gastrointestinal disorders. As these biological properties have been linked to its phenolic composition, this plant species could be an interesting source of bioactive compounds with therapeutic potential [1]. For the evaluation of chemical compounds in A. eupatoria extract, the aerial parts were harvested in the flowering period from the collection of the Scientific Practical Centre in the Field of Medicinal Plants of “Nicolae Testemițanu” State University of Medicine and Pharmacy. The dry extracts were concentrated using a rotary evaporator-Laborota 4011. The analysis was performed on Shimadzu LC-20AD chromatograph with SPD-20A UV detector [2], under the following conditions: stationary phase - Zorbax Exlipse Plus C18 (4.6x250 mm, 5 microns); 2 mobile phases: solvent mixture: methanol: water (40:60) with gradient elution and orthophosphoric acid 0.5%: acetonitrile (80:20) with isocratic elution mode; detection at wavelengths of 280, 325 and 360 nm. The solvent system that achieved optimal separation of the phenolic compounds was the mixture of 0.5% orthophosphoric acid:acetonitrile (80:20) at a wavelength of 325 nm. The extract of A. eupatoria was found to be richer in tannins (catechin-3.68%, epicatechin-2.59%); in flavonoids (rutin-1.71%, quercetin-0.199%, luteolin-0.184%, kaempferol-0.162%, apigenin-0.078%, with a lower content of hydroxycinnamic acids (chlorogenic, caffeic, cycoric and ellagic). The HPLC method with UV-VIS detection can be successfully used in the identification and quantitative determination of chemical compounds in Agrimonia eupatoria extracts for the development and standardization of new pharmaceutical forms.