The invention provides a sample pretreatment method suitable for the immunological detection of drug residues in food. The method comprises the steps of (1) treating a homogeneous tissue sample with an extracting solution and centrifuging the treated sample to collect supernate, (2) mixing the supernate collected in the step (1) with a polarity regulator to obtain a mixed solution, wherein the mixed solution and an aqueous solution are immiscible, and (3) transferring drug residues in the mixed solution obtained in the step (2) into the aqueous solution or a buffer solution with the pH value of 5.0-9.0 and directly applying the collected aqueous solution or buffer solution with the pH value of 5.0-9.0 to immunological detection and analysis. A target object extracted by the method is freeof organic solvent interference and can be directly used for immunological method detection, operation steps are simplified, the extraction efficiency and the detection efficiency are improved, and meanwhile, an immunodetection result is not influenced, and so that the method has important application value in pretreatment application of the drug residues in the food.