Anodic stripping voltammetry measures copper bioavailability for sea urchin larvae in the presence of fulvic acids

Copper speciation in the presence of fulvic acids (FAs) was studied in chemically defined seawater by square wave anodic stripping voltammetry (SQWASV). A simple complexation model assuming a single type of ligand and a 1:1 reaction stoichiometry successfully explained the measured data. A conditional stability constant of 5.80 +/- 0.07 and a complexing capacity of 610 +/- 80 micromol Cu/g FAs were obtained. This conditional stability constant would correspond to 7.23 on a free copper ion basis. The complexation kinetics was easily followed by anodic stripping voltammetry (ASV) measurements, and complexation equilibria were not reached until approximately 12 h. Coupled to speciation studies, the toxicity of copper in the presence of FAs was studied with the sea urchin Paracentrotus lividus embryogenesis bioassay. A clear protective effect of FAs was observed on Cu toxicity, and this effect could be accurately explained by SQWASV Cu speciation measurements. Additional experiments in the presence of high Cu and FA concentrations were performed, and no extra toxicity of Cu-FA complexes was observed. These results are in agreement with metal bioavailability models and also underline the usefulness of ASV for determining copper bioavailability in the presence of natural organic matter.