In this review, the application of chromatographic techniques in the separation and purification of flavonoids was summarized. The usage of column chromatography, pre-high-performance liquid chromatography, thin-layer chromatography, supercritical fluid chromatography, high-speed countercurrent chromatography and a combination of multiple chromatography methods were also reviewed. The development direction of preparative liquid chromatography columns, the optimization of solvent system for high-speed countercurrent chromatography, and the use of multi-dimensional chromatography systems were prospected.