In order to investigate the effects of biosurfactant properties , concentration and time on the removal of heavy metals from river sediment, three different bio-active agents, tea saponin, saponin and sophorolipid were selected for the study. Results indicate that washing efficiency varied among these biosurfactants. Critical micelle concentration (CMC) of biosurfactant had a significant effect on washing efficiency. Biosurfactant with the lowest CMC, i.e. sophorolipid, manifested the highest washing efficiency. In addition, washing efficiency reached its optimum at a concentration of 8%. Study on washing time to leaching effect showed that Elovich equation describe well the kinetics of washing by tea saponin, saponins and sophorolipid which means that heavy metal washing involved multi-phase diffusion. Forms of heavy metal before and after washing were also analyzed. Biosurfactant washing was effective in removing weakly-bound (ion exchangeable and carbonated-bound) heavy metal thus increased the proportion of strongly-bound ones and reduced the bioavailability of heavy metals.