The genus Eupatorium (Asteraceae) is distributed throughout tropical America, Europe, Africa, and Asia (Liu et al., 2015). Several species of this genus are found in Campos Gerais (State of Paraná, Brazil), which is a region of our studies, among them, the species Eupatorium macrocephalum Less (synonym Campuloclinium macrocephalum). This species is used in folk medicine as anti-inflammatory, sedative and in the treatment of heart diseases. The only one study on this plant reported the isolation and identification of triterpenes, steroids, the flavonoid cirsilol, and cinnamic acids derivatives (Vega et al., 2008). In continuing our studies on Eupatorium species present in Campos Gerais, in the present work we report the results of the chemical investigation of E. macrocephalum.The plant material was collected in Campos Gerais (Ponta Grossa-PR) and a voucher specimen is deposited at the Herbarium of UEPG (HUEPG 18905). The aerial parts (700 g) were extracted with methanol, and the crude extract (17g) was solubilized in methanol/water and partitioned into n-hexane, dichloromethane and ethyl acetate. Part of the dichloromethane fraction (931 mg) was successively fractionated on CC using hexane e hexane/acetone as eluent to afford the compounds EM-1 (4mg) and EM-2 (5mg) (Figure 1). The structures of isolated compounds were elucidated by analysis of their 1H and 13C NMR, HSQC, HMBC, COSY and HRESI-MS data. The 1H and 13C NMR of both compounds presented signals characteristic for sesquiterpene lactones containing the germacranolide-type skeleton (Yamada et al, 2004). The positions of the 5-hydroxytiglate group at C-8 for both compounds, as well of the acetoxyl (in EM-1) and hydroxyl (in EM-2) groups at C-9 were confirmed by HMBC analysis. The mass spectrum showed peaks at m/z 435.1610 [M+H]+ and m/z 393.1505 [M+H]+, for EM-1 and EM-2, respectively, which are consistent with the proposed structures. Compounds EM-1 and EM-2 are being described for the first time. Sequiterpene lactones with the germacranolide-type skeleton were reported to occur in E. perfoliatum, E. scabridum, E. glehnii, E. lancifolium, E. semiserratum, E. lindleyanum and E. sessilifolium (Liu et al., 2015).