The invention relates to an adsorbent for removing testosterone in blood as well as preparation and application of the adsorbent. A chitosan microsphere serves as a carrier, and a complex is formed with a zinc ion ligand on the surface of the chitosan microsphere carrier by virtue of a multi-amino spacer connecting arm. The multi-amino spacer connecting arm is hexamethylenediamine, and the contentof the zinc ligand in the adsorbent is 90-130mg/g. Appearance and physical and chemical parameters comprise: brown spherical resin, the particle size of 0.45-0.9mm, the moisture content of 50-65%, and the adsorption rate of 25-35% on the testosterone in blood. Based on an existence form of the testosterone, according to molecular structure characteristics of testosterone-binding proteins, the designed testosterone adsorbent is capable of selectively removing the testosterone from the blood, has a low rate of removing other proteins, and can be used for removing the testosterone in blood during blood perfusion.