In this study, magnetic molecularly imprinted biochar microspheres for specific adsorption of tetracylines (TCs) were prepared by Pickering emulsion polymerization using tetracyline (TC) as template and Fe₃O₄ and biochar as co-stabilizers. The adsorption properties of the imprinted microspheres for TC were studied by adsorption kinetics test, adsorption isothermal test, and selectivity adsorption test, exhibiting the excellent specific adsorption for TC with imprinting factor of 3.97. Further more, the imprinted materials were used as adsorbents for extraction and purification of TCs in real samples, and the content of TCs were detected through high performance liquid chromatography (HPLC). Under the control of the optimum conditions, the established method showed a good linearity in the range from 5 μg/L to 160 μg/L with the precision of 1.23%~9.99%, limits of detection (LOD) of 1.42~1.58 μg/L, and high recoveries of 90.6%~103.4%. This indicated that the proposed method could be applied for the accurate analysis of TCs at trace level in food.