Abstract
Migration zones of some indicator reagents were focused and separated in a polyacrylamide gel medium with a pH gradient using a disc-electrophoretic type column without discontinuous surface in the polyacrylamide gel.
Pyrocatechol Violett (PV), Xylenol Orange (XO), Thorin (T) and alizarin complexone (ALC) are expected to be separated at pH 710 in the gel by the theory of mobility-pH curve.
As pK35-values of above samples will range from 9.7 to 10.5, their mobilities (migration speed) will change sharply at about pH 9.9. So we have succeeded satisfactorily the above focusing and separations by using the pH change in gel employing the gel pH 8.9 against the electrode buffer pH 9.9.
When glycine having pK-value 9.6 is added to the electrode buffer, focusing and separation are made more efficiently. Since glycine dissociation will begin at or over pH 9.6, resulting electric resistance in electrode buffer decrease and the potential gradient in the gel medium higher, so that the migration velosity of samples will be much accelerated.
Two focused zones were observed with the XO-electrophoresis. By means of the mobility-pH curve, the one zone is ascertained to be XO and the other semi-XO (impurity in trace).
These were identified only through the focused zone.
