Ion-exchange chromatography by dicarboxyl cellulose gel

A new column packing material for ion-exchange chromatography was prepared from cellulose gel by periodate oxidation followed by chlorite oxidation to form spatially paired carboxyl groups (dicarboxyl cellulose, DCC). The carboxyl group was quantitatively introduced to spherical cellulose gel by controlling the extent of oxidation. The DCC gels were examined for their ion-exchange activity for various amines at pH of 2.5-5.5. In this pH range, aromatic amines with acid dissociation constant (pKa) below 2.7 showed no interaction with DCC gels as expected from their lack of protonation. The amines with pKa greater than 3.3, both aromatic and aliphatic, showed strong interaction corresponding to the amount of carboxyl introduced to the gel. However, these amines showed anomalous dependence on pH of the mobile phase, showing a maximum in retention factor at around pH 4. This is in contrast with the nearly constant retention factor of these amines on conventional carboxylated cellulose packing at pH greater than 4.0. The maximum retention factor at pH 4 of DCC gel was 4-5-times greater than that of conventional gel having a similar amount of carboxyls. Since pKa of dicarboxyl groups ranges 3-5 as determined by acid-base titration, the pH giving maximum retention corresponds to the pH at which one of paired carboxyls is dissociated. Possible cause of this anomaly is presented in terms of dissociation state of dicarboxyl groups and its interaction with amines.