Many phytochemical investigations have been focused recently on the antioxidant activity of herbal extracts which can be used in phytotherapy. The previous study revealed antioxidative properties of Mutellina purpurea extract, but the constituents responsible for this action were not described yet. The aim of this study was activityguided separation and identification of antioxidant compounds from M. purpurea herb. Thin-layer chromatography-2,2-diphenyl-1-picrylhydrazyl (TLC-DPPH•) assay was used to detect compounds of interest; liquid chromatography-diode array detection-mass spectrometry (LC-DAD-MS) analysis allowed to identify antioxidants. The active fractions analyzed with LC-DAD-MS contained as a main compound chlorogenic acid accompanied with p-coumaric acid, ferulic acid, three dicaffeoylquinic acids, and caffeoylferuloylquinic acid. The fast TLC-DPPH• assay with LC-DAD-MS identification enabled the accurate identification of antioxidants in M. purpurea herb, which was done for the first time.