Abstract: The present paper describes the use of high performance liquid chromatography to separate the Chinese drug venoruton.It was separated by K-1 silica gel column and was identified by UV, HPLC and TLC.The composition was found to be mainly: 7-mono-O-(β-hydroxyethyl) rutoside (HER); 4',7-di-O-(β-hydroxyethyl) rutoside 3',7-di-O-(β-hydroxyethyl) rutoside 3',4',7-tri-O-(β-hydroxyethyl) rutoside 4', 5,7-tri-O-(β-hydroxyethyl) rutoside 3', 5,7-tri-O-(β-hydroxyethyl) rutoside and 3',4',5,7-tetra-O-(β-hydroxyethyl)rutoside.The quantitative analysis indicates that it consists of about 33% of mono-HER, 37% of di-HER, 26% of tri-HER,2% of tetra-HER and 2% of rutin.