Abstract: [Objective] To determine the content of rutin, hyperoside and isoquercitrin in apocynum leaf by HPLC, and compare the content of 3 flavonoids in Apocynum leaf from different sources.[Method]The assay was performed on symmetry column (250 mm×4.6 mm, 5 μm), eluted with a gradient of acetonitrile-0.2; aqueous phosphoric acid at 35℃.The rate of flow was 1.0 mL/min, detection wavelength was 360 nm, the infection volume was 10 μL.[Result]The linear ranges of rutin, hyperoside and isoquercitrin were 0.251,1-1.255,4 μg, 0.026,6-0.265,9 μg, and 0.235,6-1.178,0 μg, and the correlation coefficients were 0.999,8, 0.999,9 and 0.999,9 respectively;The average recovery rates were 103.6;, 102.1; and 104.4;, respectively and the RSD was less than 3.8;.The content of isoquercitrin in Poacynum hendersonii(Hook.f.) Woodson was the highest,rutin in Apocynum venetum Linn was the highest ,and hypericides were detected only in Apocynum venetum Linn.[Conclusion]HPLC method can be used to determine the content of 3 flavonoids in Apocynum venetum L.There are certain rules in the content of 3 flavonoids in two kinds of Apocynum venetum, and the results can provide two methods and basis for quality control of Apocynum venetum, The content of isoquercitrin in Apocynum venetum Linn is slightly higher than that of Poacynum hendersonii (Hook.f.) Woodson, the content of hyperoside is only 0.05;, no hypericides were detected in Poacynum hendersonii (Hook.f.) Woodson, and the results can provide two methods and basis for quality control of Apocynm venetum.