Complexation and toxicity of copper and the free metal bioassay technique

The concentrations of free and total copper toxic to Daphnia and guppies were determined in inorganic media with and without addition of various concentrations of β-alanine, glycine, glutamic acid or Tris. Free copper concentrations were determined using a cupric ion electrode. Stability constants calculated for each of the detected complexes compared favourably with previously published values, with the possible exception of the Cu(OH)₂ complex. Free copper concentrations in solutions equally toxic to Daphnia were observed to vary greatly, primarily because of the toxicity of copper amino acid complexes to this organism. The copper/amino acid complexes were, nevertheless, less toxic than the free copper ions. The copper/β-alanine complex was observed to be less toxic to guppies than to Daphnia, indicating a difference in sensitivity to different copper complexes in different organisms. Copper/Tris complexes were found to be only slightly toxic to both Daphnia and guppies. A bioassay technique for determining free copper concentrations by comparing copper toxicity before and after addition of Tris was tested and verified. Although free metal concentrations can be determined from properly conducted bioassays, the variation in free metal concentration in equally toxic solutions demonstrates that free metal concentrations cannot be calculated by simply comparing metal toxicity in a test solution with toxicity of the same metal in a standard solution with known free metal concentration, unless it is known that no complexes are present in the test solution which can form toxic complexes with the metal.