A novel LC-MS/MS multi-group method for simultaneous determination of antimicrobial residues in legume-based alternative proteins

A novel liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the simultaneous analysis of three different classes of veterinary drugs—antibiotics, antifungals, and antiparasitics—in legume-based alternative protein samples, e.g. beans, peas, and nuts. The sample preparation process utilized a modified dilute-and-shoot (DnS) technique, achieving recoveries ranging from 72.08 to 108.11% for 18 of the 26 target analytes. The method demonstrated excellent repeatability (n = 12) with relative standard deviations (RSD) between 1.39 and 9.26%, and intermediate precision (over three days, n = 18) ranging from 5.76 to 19.94%. Limits of detection (LOD) and quantification (LOQ) ranged from 0.04 to 15.64 ng.g−1 and 0.10 and 47.40 ng.g−1, respectively, with good linearity. The optimized method was applied to 97 legume…