Development and validation of an LC-MS/MS method for a sensitive quantitation of livestock and poultry meat allergens

Meat allergens pose significant health risks to allergic individuals, yet highly sensitive quantification methods remain limited. In this study, a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for quantifying trace meat allergens from beef, lamb, pork, chicken, and duck in foods. Sample preparation was optimized, involving protein extraction, enzymatic digestion, and peptide purification, with five surrogate peptides from myoglobin and myosin light chain selected as quantitative markers. Multiple reaction monitoring (MRM) parameters were refined using Skyline software, and matrix effects were minimized through matrix-matched calibration and stable isotope-labeled peptides. Validation confirmed excellent specificity, linearity (R² > 0.995), limits of quantification (LOQ) of 5.0-10.0 mg/kg, apparent recoveries of 80.2 %-101.5 %, and precision (RSD < 13.8 %), with limits of detection (LOD) of 2.0-5.0 mg/kg for all five allergens. The developed method offers a robust tool for sensitive quantification meat allergen, enhancing food safety and allergen risk management.