Melampyrum barbatum Waldst. & Kit. Ex Willd. (Scrophulariaceae) has been used in traditional medicine for the treatment of rheumatic complaints and different skin diseases. In the course of our study the anti-inflammatory activity of the aerial parts of M. Barbatum was evaluated. A MeOH extract was prepared and consecutively partitioned with CHCl3, EtOAc and n-BuOH. The fractions were assayed in in vivo carrageenan-induced rat paw oedema model. The intraperitoneally administered n-BuOH phase exerted marked inhibitory effect (33.6 %, p < 0.01). Multistep chromatographic separation afforded mussaenoside and aucubine from n-BuOH fraction. Moreover, 8-epiloganin, loganic acid and mussaenoside were obtained from EtOAc fraction and apigenin, luteolin, benzoic acid and galactitol from CHCl3 fraction. These data validate the ethnomedicinal use of M. Barbatum for the treatment of inflammatory diseases and reveal that iridoids and flavonoids could be responsible for the anti-inflammatory effect of this species.