The invention discloses a preparation method of clenbuterol hydrochloride molecularly imprinted polymer microspheres. This preparation method comprises the following steps: 1) mixing nonionic surfactant solution, clenbuterol hydrochloride, porogen, initiator, inorganic nanoparticles, monoene monomer and polyene monomer, uniformly 2) heat the emulsion to react to obtain polymer microspheres; 3) wash the polymer microspheres, then soak the microspheres in acid solution, then take out the microspheres, wash and remove the acid solution, and obtain template removal 4) performing Soxhlet extraction on the template-removed microspheres, and then drying to obtain clenbuterol hydrochloride molecularly imprinted polymer microspheres. The preparation method of the present invention is simple, cost-saving, and environmentally friendly, and the prepared clenbuterol hydrochloride molecularly imprinted polymer microspheres have good uniformity, strong specific adsorption capacity, and high stability, and can be used in the fields of food safety detection and environmental restoration, etc. with broadly application foreground.