Objective：To develop an HPLC method for simultaneous determination of Rutin, Hyperoside, Isoquercitrin, Quercetin in the hebal of Apocynum venetum from different places．Methods：The separation was performed on a Shiseido C18(3μm，30㎜×100㎜) column by gradient elution with acetonitrile：methanol=10:1 as the mobile phase A 一0．1％ formic acid as the mobile phase B （0-4min，5%A→16%A；4-15min，16%→23%A；15-20min，23%A→35%A;20-22min，35%A→55%A）at a flow rate of 0.6 mL•min-1．The detection wave length of DAD was set at 360 nm，and the column temperature was 30 ℃．The injection volumn was 5μl。Results： Four flavonoids(Rutin, Hyperoside, Isoquercitrin, Quercetin) were separated at base line within 22 min with good linearity (r=0．9999) .The result of intra-day and inter-day precisions ,limits of detection and quantitation were all within the normal range. The recovery rates (n=3)were Rutin 102.0%，96.40%，103.8%；Hyperoside 98.50%，101.0%，102.9%；Isoquercitrin 98.40%，100.4%，101.4%；Quercetin 104.4%，103.1%，103.5%。respectively．The contents of four flavonoids in the hebal of Apocynum venetum from 9 different places were determined.Conclusion：The developed method is rapid，simple，accurate，reliable，and is helpful to control the quality of Apocynum venetum．