Isoform Separation of Metallothioneins by Capillary Zone Electrophoresis with Tris-Tricine Buffer in the Presence or Absence of Methanol.

A simple capillary electrophoretic method for analysis of metallothionein (MT) isoforms has been developed. rabbit liver MT isoforms are readily separated by capillary zone electrophoresis (CZE) with 150 mM tricine buffer (pH7.50) using uncoated polymide cladded fused-silica capillary column thus offering an alternative method for MT separations under original conditions. The effects of buffer concentration, buffer pH and buffer composition on the separation of MT isoforms were investigated. The effect of addition of organic solvent, MeOH, as buffer modifier on the separation was also studied. Addition of MeOH to the buffer system enhanced the separation and additional information of the heterogeneity within each isoform group was gained. Also the effect of pH on the separation is enhanced when using organic solvent modifier buffer. Enhanced information is gained by using photodiode array detection allowing recording of UV spectra of the putative isoform peaks thus offering the possibility of characterisation of the peaks obtained by comparing them to the unique characteristics of metallothionein metal-thiol bonds. The Tris-tricine method presented in this paper allows the separation of peaks that were found to need two separate runs under different conditions for separation to occur when using tris-borate buffer.

VIRTANEN Vesa;  BORDIN Guy; 

1998-06-09

JRC15926

https://publications.jrc.ec.europa.eu/repository/handle/JRC15926,