The limits of uptake of Co, Ni, and Cu by the common metallophyte, Medicago sativa, were assessed using hydroponic growth and metal uptake experiments. The influence of the growth substrate metal concentration (500 and 1000ppm) and exposure time, i.e., the time plants were exposed to the metal solution (24, 48, or 72h) was investigated. The combined roots and shoots of Medicago sativa accumulated up to 2.2wt-% Co, 2.0wt-% Ni, and 3.5wt-% Cu, when exposed to aqueous solutions containing 1000ppm Co for 48h, 1000ppm Ni for 72h, and 1000ppm Cu for 72h, respectively. The distribution of the sequestered metals was assessed using proton induced that X-ray emission spectroscopy (-PIXE), which indicated that translocation mechanism was most likely xylem loading. However, the rate of translocation of the metal from the roots to the plant stem was different for each metal, suggesting differing mechanisms for each. Collectively, these results suggest the separation and removal of the heavy metals Cu, Co, and Ni from contaminated substrates using Medicago sativa is a viable technology.