In order to study the performance of an amphoteric resin with hydroxyl group and nitrogen-containing heterocycle used for the separation of succinic acid (SA), a weak acid-weak base resin, shortly called 4-VMVH resin, the copolymerization of 4-vinylpyridine with methyl acrylate and vinyl acetate, was prepared, then the physicochemical property, performance and adsorption mechanism of the resin was explored. The results indicated that more meso- and micro-pores and less macro-pore were determined inside the resin, and the adsorption of SA on the resin was an exothermic process and could be described by Freundlich model. The distribution of adsorption site on the resin was inhomogeneous, and the isosteric adsorption enthalpy of the resin decreased with increasing the fractional loading of SA. The total acid-base exchange capacity and the hydroxyl group content of the 4-VMVH01 resin were 4.607 mmol/g and 114.21 mg KOH/g and higher than that of the 2-VMVH resin. Moreover, the resolution of SA and acetic acid (AcOH) was 0.92 while the recovery rate of SA reached to 84.67% at a ratio of the column height to its diameter of 15/1 with only hot water as eluent, and the reusability of the 4-VMVH resin basically remained constant. The interaction energy between the heterocyclic N atom and the carboxyl H atom of SA was higher than that between the heterocyclic N atom and the carboxyl H atom of AcOH. The interaction energies between hydroxyl O atom and the carboxyl H atoms of SA and AcOH were respectively -26.531 kJ/mol and -25.094 kJ/mol, which belonged to the hydrogen-bonding adsorption, and therefore the retention force of SA on the 4-VMVH resin was stronger than that of AcOH.