Content of rotenone in Derris and Tephrosia vogelii were determined by HPLC.The results in the experiment showed that the content of rotenone varied in varieties,plant growing stages(PGS) and regions of plants grown.The PGS of the highest content of rotenone in the root of D.elliptica,D.malaccensis,D.albo-rubra,and D.marginata were all in April except that D.glauca was in November.The highest content of rotenone of plant varieties is 3.13%in the root of D.albo-rubra in April.The highest content of rotenone in Tephrosia vogelii is 0.43% in the leaf and 0.25% in the stem in Baoan county Guangdong Province.The results may be useful for spreading and picking them.