Article ID: 2026.013
Temperature-responsive ion-exchange chromatography has been investigated as a promising method for the separation of ionic biomolecules. In this study, we examined the effect of methanol addition to the mobile phase on analyte elution behavior. The phase-transition behavior of poly(N-isopropylacrylamide-co-N,N-dimethylaminopropyl acrylamide-co-n-butyl methacrylate) (P(NIPAAm-co-DMAPAAm-co-BMA)) indicated that methanol addition lowered the transition temperature due to enhanced polymer dehydration. The retention times of adenosine nucleotides decreased in the presence of methanol, which was attributed to reduced electrostatic interactions caused by enhanced deprotonation of dimethylamino groups. These results demonstrate that methanol addition is an effective approach for controlling retention behavior in temperature-responsive ion-exchange chromatography.
