In this study, the adsorption and desorption properties of sulforaphane on macroporous resins (HP20, SP207, SP850, and HP2MGL) were investigated. Analysis revealed that SP850 resin was most effective in the separation of sulforaphane. The equilibrium experimental data obtained at different temperatures were well fitted to the Langmuir and Freundlich isotherms. To optimize the separation process, dynamic adsorption and desorption tests were performed with a column packed with SP850 resin. The results showed that the optimum parameters for adsorption were as follows: flow rate: 5 BV/h, pH 2, temperature: 25 degrees C; for desorption: ethanol-water (40:60, v/v), 6 BV as an eluent, flow rate: 6 BV/h. The highest purity of sulforaphane product was 85.9%, i.e., 107-fold higher than those in broccoli seeds through one run treatment on the column packed with SP850 resin under normal conditions. This indicated the high efficiency of SP850 resin in separating sulforaphane.