A procedure is developed for determining 11 β-agonists, that is, hormonal growth promoters for farm animals, using HPLC with mass spectrometric detection. The procedure includes the enzymatic hydrolysis of homogenates of organs and tissues of animals (longissimus muscle, liver, and kidney), precipitation of proteins with perchloric acid, extraction of β-agonists with a mixture of isopropyl alcohol–ethyl acetate (6: 4), purification of the extract by solid phase extraction using a Bond Elut cartridge, HPLC separation in the gradient elution mode, electrospray ionization, and mass spectrometric detection in the multiple reaction monitoring mode. The procedure is certified. The analytical range is 1.0–100.0 μg/kg.