[en] This study describes the development of a validated method for the quantification of rotenone in yam bean. The milled seeds were submitted to a Soxhlet dichloromethane extraction which allowed extracting 90% of the seeds rotenone. Elimination of the lipids was obtained via solid phase extraction. Rotenone was eluted with dichloromethane/methanol and the solution dried under vacuum and solubilised directly in methanol before injection in HPLC. The whole process was realised as much as possible protected from light and at temperatures lower than 40°C which allowed high recovery rates of spiked rotenone. Total error was used as criterion for the validation process and accuracy profiles drawn. The method allows the quantification of rotenone in yam bean seeds from 0.07% up to 1.25% (w/w). This method was applied to the quantification of rotenone in the seeds of several accessions of Pachyrhizus erosus and P. ahipa. The results range from 1.13 to 2.76 mg/g dry material.