Separation mechanism of pullulan solution-filled capillary electrophoresis of sodium dodecyl sulfate-proteins.

The separation mechanism of capillary electrophoresis of sodium dodecyl sulfate (SDS)-proteins using pullulan with a molecular mass range of 50,000-100,000 as a separation matrix was investigated. The pullulan solution was filled into fused-silica capillaries whose inner walls were coated with linear polyacrylamide through chemically stable Si-C linkages. Baseline separations of SDS proteins were achieved at concentrations ranging from 3-10% w/v of pullulan. The entanglement threshold of pullulan solution was found to be around 0.5% w/v, indicating migration of SDS-proteins through an entangled pullulan network. Ferguson plots exhibited a linear relationship between log mobility and pullulan concentration. Linear relationships were also obtained for double logarithmic plots of the electrophoretic mobility and protein molecular mass. These results show that the separation is based on mass discrimination in accordance with the Ogston theory.