Abstract: OBJECTIVE  To analyze the prototype components and metabolites of aqueous extract of Sanguisorba officinalis L. by UHPLC-Q-TOF/MS in feces, urine and plasma of rats and explore its metabolic pathways.METHODS  Chromatographic separation was performed on the Thermo Acclaim^TM RSLC 120 C₁₈ column (3.0 mm×100 mm, 2.2 μm). The mobile phase was consisted of 0.1% formic acid in water (solvent A) and acetonitrile (solvent B). The sample injection volume was 3 μL and the flow rate was set to 0.4 mL·min^-1. The column temperature was maintained at 40 ℃. The detection was carried out over a mass range of m/z 50-1 500 and equipped with an electrospray ionization (ESI) source operating in positive mode and negative mode. Chemical composition on Sanguisorba officinalis L. and its major metabolites were identified by comparing the differences in feces, urine and plasma between adminstration samples and blank samples in rats.RESULTS  After intragastric administration of the aqueous extract of Sanguisorba officinalis L., 69 exogenous components were identified from rat plasma, urine and feces, including 24 prototype components and 45 metabolites. 24 prototype components included 16 phenolics, 7 triterpenes, and a monoterpene glycoside. The metabolic pathways of the aqueous extract of Sanguisorba officinalis L. in rats mainly included reduction, methylation, dehydration, oxidation, sulfation, and glucuronidation.CONCLUSION  The identified prototype drugs and the metabolites can supply a clear material basis for the further study of efficacy of Sanguisorba officinalis L..