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Aim The study focused on identification and antidepression active verification of constituents from Angelica Sinensis Radix contrubuting to Xiaoyaosan based on UPLCPDA guided isolation technique.Methods The UPLCPDA chromatogram of Xiaoyaosan was compared with that of Angelica Sinensis Radix.The relative retention time of each peak and the Uhraviolet spectra providing by PDA were used in the analyses.Constituents were isolated from Angelica Sinensis Radix under the guidance of UPLCPDA investigation.Structures of the isolates were elucidated by NMR techniques.Antidepression effect was evaluated on glutamateinduced neurons.Results Five marker peaks of Xiaoyaosan fingerprint were belong to Angelica Sinensis Radix, and they were determined as coniferyl ferulate (Ⅰ), Ebutylidenephthalide (Ⅱ), ligustilide (Ⅲ), Zbutylidenephthalide (IV), 14Acetoxy12senecioyloxytetradeca2E ,BE, 10Etrien4,6diyn1ol (V).Compound V was isolated from the plants of Umbelliferae for the first time.Treatment with compound Ⅰ, Ⅱ, V can protect PCl2 and SHSY5Y cell from glutamateinduced cytotoxicity.Antidepression bioactivity of compound Ⅰ was first investigated.Conclusion UPLCPDA guided isolation technique was confirmed to be a rapid and accurate method to identify the main active constituents from Angelica Sinensis Radix contrubuting to Xiaoyaosan.