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  • Separation of metallothionein isoforms with capillary zone electrophoresis using an uncoated capillary column : Effects of pH, temperature, voltage, buffer concentration and buffer composition


Metallothionein(MT) isoforms, sulphydryl rich proteins with a high affinity for essential as well as toxic trace metals, are readily separated by capillary zone electrophoresis (CZE) with an uncoated polyimine cladded fused silica capillary using TRIS borate buffer. Investigated metallothionein samples were rabbit liver MT, rabbit liver MT-1, rabbit liver MT-2 and horse kidney. MT. Effect of temperature, buffer pH, buffer concentration, buffer composition and running voltage on the separation efficiency was investigated. Improved separation efficiency compared to those methods mentioned in the literature using uncoated fused silica capillary was obtained allowing separation of peaks which were earlier comigrating. The used running conditions were: voltage 11 kV, temperature 20 C, buffer TRIS 110 mM - borate 110 mM, pH 6.90. Under the experimental conditions horse kidney MT exhibited 5 putative isoform peaks and rabbit liver MT 9-10 peaks. Interestingly, experiments indicated that rabbit liver MT contains one major component which is found neither in isoforms MT-1 nor MT-2 maybe indicating that the improved separation efficiency made it possible to detect a new group or structure. Based on our initial evaluation, CZE with uncoated fused silica capillary using TRIS borate buffer appears to be a very useful analytical method for the separation of metallothionein isoforms.

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Bibliographic Reference: Article: Journal of Chromatography A
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