Medicinal plants reveal their therapeutic effects due to the high amount of phenolic compounds confined abundantly in roots, fruits, leaves, and flowers. Rosa heckeliana Tratt, as a member of Rosa, was recently discovered as one of the important medicinal plants used in folk medicine. In this study, the therapeutically notable Rosa heckeliana was selected for the analysis of important phenolic components particularly confined in the roots. The roots were first extracted in methanol, then phenolic constituents were partitioned according to polarity by fractionation method. The method was also supported by the total amount of phenolic compounds. The highest total phenolic content as gallic acid equivalent (8.8 +/- 0.19 mu g/mg) was obtained in the ethyl acetate fraction. Hence, that fraction was further analyzed for the identification of phenolic constituents by employing high performance liquid chromatography equipped with diode array and fluorescence detectors. Structural information on the compounds was confirmed by high-resolution mass spectrometry. Accordingly, catechin, caffeic acid, epicatechin, p-coumaric acid, scopoletin, ellagic acid, and quercetin were identified in the ethyl acetate fraction. Catechin, epicatechin, and scopoletin revealed fluorescent characteristics. To the best of our knowledge, this is the first detailed effort for the fractionated analysis of phenolic constituents in the roots of Rosa heckeliana.