Extracellular polymeric substances (EPSs) excreted by the bacterium Sinorhizobium meliloti and associated Ca, Cd and Pb were characterised by asymmetrical flow field-flow fractionation coupled with UV spectrophotometry and inductively coupled plasma mass spectrometry in terms of molar-mass distributions, number- and weight-average molar masses and polydispersity index. Two major populations with weight-average molar masses of 74 × 10^3 and 1.35 × 10^6 g mol^–1 were obtained for the EPS. Characterisation of the whole EPS–metal interactions evidenced the preferential binding of Ca and Cd to the low molar mass fraction, whereas Pb associated mainly with the high molar mass (HMM) fraction. Comparison with the EPS produced by exoY-mutant, deficient in HMM-EPS excretion, confirmed the preferential binding of Pb to the high molar mass fraction. Enrichment of the EPS with increasing metal concentrations induced the formation of aggregates, which was most pronounced in the presence of 10^–4 mol L^–1 Pb.