Objective To develop an efficient detection method based on the Waters TQ-XS triple quadrupole liquid chromatography-mass spectrometry (LC-MS/MS) system for identifying cases of duck meat being substituted for beef or lamb.Methods Using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), specific peptide ion pairs of beef, lamb, and duck were screened and validated. A qualitative and quantitative analysis method was established under multiple reaction monitoring (MRM) mode, with optimization of sample pretreatment and mass spectrometry conditions.Results The method showed good linearity (R²>0.99), high sensitivity, and a minimum limit of quantification of 0.5%. Detection of samples with different adulteration ratios demonstrated that the method could accurately and rapidly identify and quantify duck meat substituted for beef or lamb. Results from actual sample testing were consistent with those obtained by PCR.Conclusion This method demonstrates excellent performance in sample processing and sensitivity, and is suitable for the rapid identification and quantitative analysis of duck meat adulterated into beef and lamb products. It can provide effective technical support for meat quality control, market supervision, and prevention of meat adulteration.