The invention provides a desorption regeneration method of a hydrophobic macroporous polymer adsorbent adsorbed with butanol. The method disclosed herein comprises the steps of using water-soluble low-boiling polar solvent and water to successively elute the hydrophobic macroporous polymer adsorbent adsorbed with butanol. According to the invention, after a solution containing butanol (e.g. butanol fermentation liquor) is adsorbed by the hydrophobic macroporous polymer adsorbent, by using the desorption regeneration method disclosed herein, the full dissolution and desorption of the adsorbed butanol in the adsorbent can be realized; in addition, the regeneration treatment can be directly carried out on the adsorbent on a fixed bed where the butanol fermentation liquor is adsorbed, and the adsorption process of the next stage can be directly conducted without taking out the adsorbent from the fixed bed, thus a lot of time is saved, and the regeneration efficiency is raised. The method disclosed herein has the advantages of simple process, short separation time, easy, rapid and complete desorption and regeneration, low investment of devices, low pollution, and low energy consumption, and is easy for large scale production.