Different parts of Ruta chalepensis L. medicinal plant and soil samples collected from three zones in Jordan (Irbid, Al-Mafraq and Ma'an) were analyzed for determination of Pb, Cu, Zn, Cd, Ni and Fe by atomic absorption spectrometry. Heavy metal levels in washed and unwashed parts of R. chalepensis were evaluated and compared statistically. Results show that there was a variation in heavy metal concentrations related to washed and unwashed parts of R. chalepensis in the three zones. Moreover, Pb, Zn, Cu, Cd, and Ni were concentrated in the following order:Two standard reference materials of plant (SRM 1790a; spinach, CRM 281; rye grass) and a standard reference material of soil (GBW 07406) were examined to validate the method used.Results indicate that high recoveries were obtained. Significant variation existed in metal content in different parts of Ruta chalepensis L. for the various locations (P ≤ 0.05).