A simple method for the determination of isoelectric points of ampholytes with closely spaced pKa values using pressure-mediated capillary electrophoresis.

A simple method, based on a modified version of pressure-mediated capillary electrophoresis (PreMCE) has been developed for the determination of the isoelectric points of ampholytes which have closely spaced pKa values. This new pI-determination PreMCE method (i) can be easily executed on most commercial capillary electrophoresis instruments; (ii) it can use small, impure samples, (iii) unlike isoelectric focusing methods in natural pH gradients, it does not require a linear pH gradient, and (iv) it eliminates the pI errors that are due to chromatographic retention on the walls of the separation chamber.