The present invention provides a method for desorbing and regenerating a butanol-adsorbing hydrophobic macroporous polymer adsorbent, comprising: successively eluting the hydrophobic macroporous polymer adsorbent with butanol adsorbed therein using a water soluble low-boiling-point polar solvent and water. After a butanol-containing solution, such as a butanol fermentation liquor, has been adsorbed by a hydrophobic macroporous polymer adsorbent, the butanol adsorbed in the adsorbent can be thoroughly dissolved and removed by the desorption and regeneration method of the present invention; furthermore, the adsorbent can be directly regenerated to recover its adsorption capability on a fixed bed adsorbing the butanol fermentation liquor, wherein the next stage of adsorption process can be directly entered without taking the adsorbent out from the fixed bed; therefore it saves a great deal of time and improves the regeneration efficiency. The method provided in the present invention has a simple process, a short separation time, easy, fast and complete desorption and regeneration, low equipment investmentand pollution, and reduced energy consumption, and therefore production is easy on a large scale.