ETHNOPHARMACOLOGICAL RELEVANCE: 'Ershiwuwei Shanhu' pill (ESP), a classical and famous prescription of traditional Tibetan medicine, has a long history of empirical clinical use for the treatment of cerebrovascular and neurological diseases, but the absence of scientific evidence for its effect restricted its clinical application and further development.MATERIALS AND METHODS: The methodology of plasma pharmacochemistry was adopted to analyze the potentially bioactive components in ESP extracts. A method based on UPLC-DAD/Q-TOF-MS was established to identify herb components in ESP extracts and analyze the absorbed components of ESP and their metabolites in rat plasma, brain, heart, liver and kidney samples after oral administration of ESP extracts. RESULTS: A total of 61 herb components were detected and identified in ESP extracts, while 35 absorbed components-including 19 prototype compounds and 16 metabolites-were discovered as potentially bioactive components in rat plasma and tissues by comparative analysis of the UV and MS chromatograms of ESP extracts, blank biosamples and dosed biosamples. CONCLUSIONS: The potentially bioactive components of ESP extracts identified from rat plasma and tissues provide useful information for further study of the pharmacology and mechanism of action of ESP.