ObjectiveTo analyze the constituents and metabolites of Radix Saposhnikoviae(RS) in rat plasma and urine by rapid-resolution liquid chromatography-time of flight mass spectrometry (RRLC-TOF/MS), so as to explore the active ingredients and metabolites of RS in vivo.MethodsThe separation was performed on a Angilent Zorbax Extend-C18 (5 μm, 250 mm×4.6 mm id) column, with a methanol-water mobile phase system used for gradient elution.Time-of-flight mass spectrometer (TOF/MS) was applied for qualitative analysis under positive ion mode.Based on the accurate molecular weight of TOF/MS detection and the compound list of RS established previously, the constituents and metabolites of RS in different matrix in vivo were identified.ResultsSix constituents of RS were identified in the plasma:sucrose, prim-O-glucosylcimifugin, cimifugin, nodakenetin, 5-O-methylvisamminol, and 3’-O-i-butyrylhammaudol.Eight constituents were identified in the urine:prim-O-glucosylcimifugin, divaricatacid, cimifugin, 4’-O-glucosyl-5-O-methylvisamminol, (3S)-2,2-dimethyl-3,5-dihydroxy-8-hydroxymethyl-3,4-dihydro-2H,6H-benzo-\[1,2-b:5,4-b’\]dipyran-6-one, 5-O-methylvisamminol, sec-O-β-D-glucosylhammaudol, and wogonin.Two metabolites were identified in the urine:glucuronide of cimifujin and an isomer of it.ConclusionThe present method is reliable and effective for identifying compounds of RS in vivo, and it can provide a reference and evidence for the further pharmacodynamics experiments.