A high performance liquid chromatography(HPLC) was established for determination of the characteristic spectra as well as the multi-index contents of substance benchmarks for classical prescription Danggui Sini decoction,which elucidated the critical quality attributes and improved the quality control system.The chromatographic analysis was performed on a Pomenex Luna C18 column(250 mm×4.6 mm,5 μm) at 30 ℃.The gradient elution was carried out with a linear gradient system consisted of acetonitrile-0.05% phosphoric acid water solution.The signal was monitored at 230 nm and 290 nm.The flow rate was maintained at 1.0 mL·min-1.The similarity of characteristic spectra for 15 batches of substance benchmarks was more than 0.89,while 45 common peaks were assessed,and 7 common components were identified as paeoniflorin,ferulic acid,liquiritin,ammonium glycyrrhizinate,asarinin,cinnamaldehyde and ligustilide.The average yield of dry extract for 15 batches of substance benchmarks was 15.00%.The contents of paeoniflorin,ammonium glycyrrhizinate,asarinin,cinnamaldehyde and ligustilide in the 15 batches of substance benchmarks were 8.226-13.46，1.929-6.535，0.020 02-0.034 30，0.076 61-0.543 3 and 0.075 94-0.140 6 mg·g-1,respectively.This study provides a scientific basis for the quality control of substance benchmarks as well as the development of compound preparation of Danggui Sini decoction.