Abstract: A high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was established for the determination of amitraz and its metabolites in eggs, and the residue dissipation patterns in eggs were investigated. Laying hens were sprayed with 0.05% and 0.1% amitraz solutions, and the low-concentration group received a second administration one week later. The residual levels of amitraz and its metabolites in eggs were measured at different time intervals after the administration. The results showed that the residue concentrations of amitraz and its metabolites in eggs increased initially, and reached the peak on the 3rd day. The residues of the low-concentration group did not exceed the European Union's temporary maximum residue limit, while the high-concentration group required a withdrawal period of 9.38 days. This study provides a scientific basis for the safe use of amitraz and for the food safety of laying hens.