Spectrophotometric detection of labile zinc(II) released from metallothionein : A simple method to evaluate heavy metal toxicity(METHODS)

概要

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A new and simple method based on the mechanism of detoxification of metallothionein was developed by using a water-soluble porphyrin and Zn(II)-bound metallothionein for evaluating heavy metal toxicity. Labile Zn(II) ions were released when toxic metal ions such as Cu(II), Pb(II), Bi(III), Cd(II), Hg(II), Co(II), Ag(I), and Ni(II) bound to Zn(II)-bound metallothionein. The water-soluble porphyrin, 5,10,15,20-tetraphenyl-21H,23H-porphinetetrasulfonic acid, a chromogenic reagent that is highly sensitive to Zn(II), formed a complex with the labile Zn(II) ions. The absorption change at 423nm resulting from the formation of the Zn(II)-porphyrin complex was used to evaluate the toxicity of sample solutions containing different metal ions. The absorption change was well correlated with the toxicity, which was evaluated by a bioluminescence inhibition assay using the bioluminescent bacteria Vibrio fischeri. This observation indicated that the absorption change determined by our method was a good indicator of heavy metal toxicity. The proposed method was more sensitive than conventional bioassays and could be used to detect metal toxicity at submicromolar concentrations of toxic metal ions.