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A Mercury saturation assay for measuring metallothionein in fish
Author(s) -
Dutton Michael D.,
Stephenson Malcolm,
Klaverkamp Jack F.
Publication year - 1993
Publication title -
environmental toxicology and chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.1
H-Index - 171
eISSN - 1552-8618
pISSN - 0730-7268
DOI - 10.1002/etc.5620120707
Subject(s) - metallothionein , chemistry , mercury (programming language) , trichloroacetic acid , chromatography , metal , saturation (graph theory) , centrifugation , isotope dilution , rainbow trout , dilution , mercure , biochemistry , analytical chemistry (journal) , mass spectrometry , fish <actinopterygii> , biology , fishery , organic chemistry , physics , mathematics , combinatorics , computer science , gene , programming language , thermodynamics
An accurate, rapid, sensitive, and simple method using mercury saturation for quantifying metallothionein (MT) is described. A complex solution (“pseudocytosol”) of enzymatic and non‐enzymatic thiols, including rabbit liver MT‐II, and supernatants from homogenized samples of rainbow trout liver were incubated in the presence of 203 Hg in 10% trichloroacetic acid Excess Hg was bound to and removed by chicken egg albumin, which denatured on contact with the acidic assay medium. After centrifugation, MT labeled with 203 Hg remained in the TCA supernatant and was estimated using known stoichiometry for Hg‐MT binding. A dilution series was used to establish that nonspecific metal binding, a common problem with other metal saturation assays, is negligible. Analysis of hepatic MT with high Cu content from rainbow trout demonstrated virtually complete displacement of Cu, Cd, and Zn by Hg. When compared to other metal‐saturation assays developed for vertebrates, this method requires the least number of technical steps, and one‐third or less of total preparatory and analytical time.