In this study, lead bioaccumulation by the living free-floating aquatic macrophyte Eicchornia crassipes in different hydroponic conditions with variations in phosphorus and lead concentrations was investigated. A set of growth experiments in hydroponic media doped with lead and phosphorus within a wide concentration range was performed for 32 days in a greenhouse. All experiments were carried out with periodic replacement of all nutrients and lead. The concentration of lead and nutrients in biomass was determined by synchrotron radiation-excited total reflection X-ray fluorescence. By increasing the lead concentration in the medium, a reduction in biomass growth was observed, but a higher phosphorus retention in roots and leaves was shown at lower lead concentrations. In addition, an increase in the amount of bioaccumulated lead and phosphorus in roots was observed for higher lead and phosphorus concentrations in the medium, reaching saturation values of 4 mg Pb g−1 and 7 mg P g−1, respectively. Four non-structural kinetic models were tested, to represent the bioaccumulation of lead and phosphorus in roots. Pseudo-second order and irreversible kinetic models described the lead bioaccumulation data well, however, an irreversible kinetic model better fitted phosphorus uptake in roots.