Active components of herbal medicines usually become trace in the body, owing to absorption/distribution barriers and/or biotransformation hurdles. The goal of this study was to detect and identify angular-type pyranocoumarins (APs) in biological samples (plasma, urine and feces) obtained from rats after oral intake of Peucedani Radix extract (PRE), using ultra-high performance liquid chromatography-tandem mass spectrometry. Initially, 11 reference compounds were used to identify the pyranocoumarin prototypes. Predictions for the in vivo compounds were performed on the basis of chemical profiling for PRE and on the metabolic pathways of the APs as characterized in our previous reports. The predictions subsequently gave rise to a predictive multiple ionmonitoring-information dependent acquiring-enhanced product ion (pMIM-IDA-EPI) procedure. With the help of reference compounds, a number ofpyranocoumarins were unambiguously identified in plasma, urine and fecal samples (10, 8, and 8, respectively); the chemical structures of an additional 21 in plasma (8 prototypes and 13 metabolites), 10 in urine (6 prototypes and 4 metabolites) and 8 in feces (4 prototypes and 4 metabolites) were tentatively assigned based on their mass spectral profiles. Collectively, the method adopting the pMIM-IDA-EPI system was demonstrated to be reliable and can be introduced as an alternative procedure for rapid characterization of herbal medicine-derived components in biological samples.