Abstract: A hormone mass spectrometric detection method based on antibody-coupled enrichment technology. The method includes the following steps: coupling antibodies and magnetic beads, adding a high-concentration methanol solution, removing endogenous hormone small molecule compounds carried on the antibodies by vortex, then restoring the activity of the antibodies to obtain antibody magnetic beads that can enrich hormone small molecule compounds. The antibodies are immobilized by magnetic beads and cooperate with the 80% methanol aqueous solution to elute hormone small molecule compounds, and then the hormone small molecule compounds are tested by mass spectrometry, thereby improving the specificity of test.