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A sensitive LC/MS method was established to investigate the in vivo metabolism of GuanXin II prescription, a five-component Chinese herbal medicine formulation. Rat plasma, bile, urine, and feces were collected and analyzed following oral administration of the water decoction. A total of 50 compounds were identified, including 17 prototypes and 33 metabolites underwent methylation, oxidation, hydrolysis, sulfate conjugation, glucuronide conjugation, and glutathion conjugation. In addition, the component herb of the formulation from which the metabolites were derived was also identified. Among the five component herbs, Rhizoma Chuanxiong, Flos Carthami, and Lignum Dalbergiae Odoriferae were actively metabolized, contributing 26 metabolites and 2 prototypes, while Radix Salviae Miltiorrhizae and Radix Paeoniae Rubra underwent less biotransformation, yielding 7 metatbolites and 15 prototypes. This is …