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<sup>-1</sup> and 0.10 and 47.40 ng.g<sup>-1</sup>, respectively, with good linearity. The optimized method was applied to 97 legume samples (primary and processed products) originating from domestic and international markets. Occurrence analysis revealed that all analytes monitored were below the detection limits, suggesting that antimicrobial contamination in legumes-based alternative protein products is low within the Association of Southeast Asian Nations (ASEAN) region.
Authors
- Chanita Boonkanon (ORCID: https://orcid.org/0000-0002-8104-5133)
- Awanwee Petchkongkaew
- Brett Greer (ORCID: https://orcid.org/0000-0001-5039-0244)
- Umaporn Uawisetwathana (ORCID: https://orcid.org/0000-0001-6782-7617)
- Robin Philp
- Nitsara Karoonuthaisiri (ORCID: https://orcid.org/0000-0003-0838-8399)
- Kawisara Siwarak
- C.T. Elliott
- Nazmi Waesoh
- David Bradley
Institutions
- Thammasat University (TH)
- Queen's University Belfast (GB)
- National Science and Technology Development Agency (TH)
- Queens University (BD)
- Agilent Technologies (Australia) (AU)
- Agri Food and Biosciences Institute (GB)
- SEGi University (MY)
- National Center for Genetic Engineering and Biotechnology (TH)
Publication Details
- Journal
- npj Science of Food
- Published
- 2026-01-03
- DOI
- https://doi.org/10.1038/s41538-025-00678-3
- Primary Topic
- Pesticide Residue Analysis and Safety
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- European Commission
- National Science and Technology Development Agency
- National Center for Genetic Engineering and Biotechnology
- Thailand Science Research and Innovation
- HORIZON EUROPE Framework Programme