Separation and quantitation of metallothionein isoforms from liver of untreated rats by ion-exchange high-performance liquid chromatography and atomic absorption spectrometry |
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Authors: | Karl N stelbacher, Manfred Kirchgessner,Gabriele I. Stangl |
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Affiliation: | Institute for Nutritional Sciences, University of Technology of Munich, Hochfeldweg 2–6, 85350 Freising-Weihenstephan, Germany |
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Abstract: | A sensitive method for determination of metallothionein (MT) isoform levels in rat liver by ion-exchange high-performance liquid chromatography and atomic absorption spectrometry was developed. Critical steps in sample preparation, like MT extraction, MT saturation with Cd and protein separation, were optimized. This method is capable of measuring levels of 2.0 μg/g liver for metallothionein-1 (MT-1) and 1.3 μg/g liver for metallothionein-2 (MT-2), respectively, with a high recovery of 103% on average. The method described, thus, proved suitable for analyzing metallothionein isoform concentrations even in untreated animals. The ratio of MT-1 to MT-2 was found to be 1:1 on average. MT decomposition during storage was very high in whole livers, but could be reduced by about 80% when extracted liver samples were used. |
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Keywords: | Metallothionein isoforms Ion-exchange HPLC Liver Rat |
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