In Turkish traditional medicine, Tripleurospermum species have been used for the treatment of inflammatory diseases. The present study was designed to investigate the anti-inflammatory potential and fatty acid composition of the extracts prepared from Tripleurospermum parviflorum (Willd.) Pobed. and T. tenuifolium (Kit.). Anti-inflammatory activity was assessed by using carrageenan-, and serotonin- induced hind paw edema and acetic acid-induced increase in capillary permeability models. The fatty acid compositions of the plants were investigated by gas chromatography (GC). EtOAc extracts of T. tenuifolium and T. parviflorum exerted notable inhibitory effect in the all in vivo anti-inflammatory activity models tested. Generally, C 16:0 palmitic acid and C 18:2 linoleic acid were found to be the major fatty acids in two species. Saturated fatty acids (SFAs) were found in higher amounts than monounsaturated fatty acids (MUFAs) and polyunsaturated fatty acids (PUFAs) in two species. SFAs were determinated at 63.15% and 58.68% in T. tenuifolium and T. parviflorum, respectively. The high content of linoleic acid and palmitic acid may be primarily responsible for significant anti-inflammatory activity. The present study confirms the anti-inflammatory activity of T. parviflorum and T. tenuifolium. Further phytochemical and biological activity studies are needed for the determination of the active principle/s and anti-inflammatory activity mechanism.