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Xue-Fu-Zhu-Yu-Tang (血府逐淤〓) is a famous traditional Chinese medicine (TCM) formula for treating cardiovascular disease and related ailments in China for centuries. To profile the phytochemical constituents of the formula, an HPLC-DAD-ESI-MS analytical method has been developed to separate and determinate the medium- or non-polar fraction of the decoction, which has been demonstrated potency to lower the serum total triglyceride concentration, strongly decrease the TXA_2/PGI_2 ratio and attenuate production of proinflammatory cytokines in high cholesterol-fed rats. By comparing their retention time, UV and MS data with those obtained from the authentic compounds, ferulic acid (1), naringin (2), neohesperindin (3), naringenin (8), marmin (13), senkyunolide A (14), dehydrosafynol (16), safynol (17) and Z-ligustlide (18) are unequivocally determined. Moreover, additional thirteen compounds are tentatively identified as senkyunolide I (4), senkyunolide H (5), poncirin (7), benzoylpaeoniflorin (10), (Z)-6, 7-epoxyligustilide (11), senkyunolide G (12), 2-methoxy-safynol (15), cnidilide (19), tangeritin (20), saikosaponin b_2 (21), 2'-0-acetylsaikosaponin b_2 (22), saikosaponin b_1 (23) and auraptene (24), according to the comparison of their UV and MS data with the published data. The present study provides an approach to rapidly characterize bioactive constituents in TCM formulae.