FACTORS INFLUENCING THE SEPARATION OF METALLOTHIONEINS BY CAPILLARY ZONE ELECTROPHORESIS

The factors influencing the separation of metallothioneins (MTs) by ca pillary zone electrophoresis (CZE) were studied using untreated fused- silica capillaries. A comparison was made between the various buffers of 20 mM Na2B4O7, 10 mM Na2B4O7-10 mM Na2HPO4 and 20 mM Na2B4O7-10 mM Tris for the separation of MTs in terms of migration time and UN-absor bance under the various applied voltages. The migration times for MT-1 and MT-11 decreased with increase in the applied voltage and column t emperature. However, no significant change in UV absorbance was observ ed. When Na2B4O7-Tris buffer was chosen for the separation of MTs, the migration time, UV absorbance, theoretical plates and resolutions als o increased with increasing buffer concentrations. The effect of buffe r pH on migration time was relatively complicated. Under the optimal o perating conditions the standard MTs were well separated. Determinatio n of MTs in the real samples failed due to the adsorption of other pro teins onto the capillary walls.