The present invention provides a chromatographic method for separating and purifying recombinant human lactoferrin from transgenic rice seeds, comprising the following steps: (1) by using transgenic rice seeds with recombinant human lactoferrin as a raw material, using an extraction buffer to prepare an rLF crude extract and (2) separating and purifying the rLF crude extract by cation exchange chromatography, to obtain a purified rLF target object. The chromatographic method further comprises a step of regenerating resin.