Previously performed studies have shown that alfalfa shoot biomass can bind an appreciable amount of nickel(II) and chromium(III) ions from aqueous solution. Direct and indirect approaches were applied to study the possible mechanisms involved inmetal binding by the alfalfa biomass. The direct approach involves investigations of the metal-bound alfalfa shoot biomass by X-ray absorption spectroscopic analysis (XANES and EXAFS). Results from these studies suggest that nickel(II) and chromium(III)binding mostly occurs through coordination with oxygen ligands. Indirect approaches consist of chemical modification of carboxylate groups that have been shown to play an important role in metal binding to the alfalfa biomass. An appreciable decrease inmetal binding resulted after acidic methanol esterification of the biomass, indicating that carboxyl groups are entailed in the metal binding by the alfalfa biomass. In addition, base hydrolysis of the alfalfa biomass increased the binding of thesemetals, which further indicates that carboxyl groups play an important role in the binding of these metal ions from solution. Therefore, by combining two different techniques, our results indicate that carboxylate groups are the major ligands responsiblefor the binding of nickel(II) and chromium(III) by alfalfa biomass.
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