Cook Your Own Ampholytes For Use In Isoelectric Focusing And Chromatofocusing

Abstract

Isoelectric focusing and chromatofocusing are techniques to separate amphoteric compounds based on their isoelectric points. The commonest used compounds to generate a pH gradient in these techniques are ampholytes. The prohibitively expensive cost of carrier ampholytes limits the application of these techniques in less advantaged parts of the world. A simple and inexpensive method is described in this paper to prepare a broad pH range (pH 3 to 11) ampholytes for use in isoelectric focusing or chromatofocusingin any routine laboratory. The method involves crosslinking polyethylene polyamine to create complex cross-linked polyaminesin the first step. Whereas, the second step involves reacting the cross-linked polyamines with an unsaturated carboxylic acid to produce ampholytes. [Kalaria T Natl J Integr Res Med, 2021; 12(3):105-109]