High molecular weight poly(2-vinylpyridine) (P2VP) was characterized by temperature gradient interaction chromatography (TGIC). In TGIC, the column temperature is varied during the elution to control the solute retention. Anionic polymerized homo poly(2-vinylpyridine) samples were successfully separated by TGIC with much higher resolution than size exclusion chromatography. An efficient interaction chromatography separation condition was established for the P2VP characterization using bare silica column, and the molecular characteristics of P2VP obtained from the temperature gradient IC analysis were compared to SEC. The MWD of anionic polymerization P2VP was found much narrower than the values commonly measured by SEC like other polymers prepared by anionic polymerization. The results show that amine-containing polymers can be separated ith high-resolution by IC.