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An ethyl acetate extract from the barks of the ethnic Chinese medicine <i>Daphne tangutica</i> Maxim. exhibited antihepatocellular carcinoma activity against HepG2 and Hep3B cell lines. By using high-performance liquid chromatography based activity profiling in combination with offline liquid chromatography with mass spectrometry and NMR analysis, we rapidly identified ten major components of the extract, including seven active principles, coumarins (<b>1-4</b>) and biscoumarins (<b>7, 8, 10</b>), along with three inactive flavonoids (<b>5, 6, 9</b>). This study demonstrated that our combined protocol can be used as an important strategy for chemical profiling, dereplication as well as the identification of bioactive compounds from herbal medicines.