Xiao-Yao-San-Jia-Wei (XYSJW) is a commonly prescribed formulation for depression and anorexia in the Jiang Su Province Hospital of Chinese Medicine. Unfortunately, the proper dosage of this formulation is still unclear due to its limited chemical and pharmacokinetic profiles. Thus, in the present study, a sensitive, precise, and rapid procedure for the identification of absorbed compounds (Cs) in the plasma of depressed rats together with a pharmacokinetic analysis was established with the help of ultra-flow liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UFLC-Q-TOF MS/MS) and ultra-flow liquid chromatography coupled with electrospray ionization triple quadrupole tandem mass spectrometry (UFLC-QQQ MS/MS). Based on the characteristic fragmentation, neutral loss, mass defect filter, relevant literature and reference standards, 225 Cs in the XYSJW extract and 20 Cs in the plasma of the depressed rats were tentatively recognized UFLC-Q-TOF MS/MS and UFLC-QQQ MS/MS. Then, the 12 major absorbed Cs in the depressed rats after oral XYSJW administration were chosen to further investigate its pharmacokinetic profile by UFLC-QQQ MS/MS. This study provides a systematic approach for the rapid and qualitative analysis of absorbed Cs in depressed rats and investigating the pharmacokinetics of XYSJW. More importantly, our work provides key information on the chemical and pharmacokinetic profiles of XYSJW and , which may benefit its therapeutic efficacy and further pharmacological studies involving this formulation.