To establish the HPLC fingerprint of Sinopodophyllum hexandrum,and to determine the contents of four constituents.JADE-PAK ODS（4.6 mm×250 mm,5 μm）column was used.Acetonitrile (A)-0.04% formic acid (B) was used as the mobile phase for gradient elution.The flow rate was 1.0 mL/min.The eluate was monitored by UV absorption at 254 nm.The column temperature was 30 ℃,and the injection volume was 10 μL.Results:HPLC fingerprint of S. hexandrum was established,and the similarity of 12 batches of S. hexandrum samples were compared.And 11 common peaks were identified,among which 4′-demethylpodophyllotoxin,quercetin,kaempferol and podophyllotoxin were identified.A method for simultaneous determination of 4′-demethylpodophyllotoxin,quercetin,kaempferol and podophyllotoxin was established,and the linear relationship between the peak area and the range of the injection volume was good,respectively.The RSD of precision,stability,repeatability,and sample recovery all meet the requirements.The content determination results showed that the contents of four main constituents in S. hexandrum from different regions were significantly different,and the contents of four main constituents in S. hexandrum from Tibet reached the highest level.This study established a method for HPLC fingerprint and multi-component content determination of S. hexandrum from different origins.This method has strong specificity and good repeatability,providing a reference for improving the quality control of S. hexandrum.