Development and Evaluation of a Preliminary Screening Assay for Antibiotic Residues in Meat

• Published in: Applied biochemistry and biotechnology 2021-04, Vol.193 (4), p.1129-1146
• Main Authors: Fan, Wei, Gao, Xiao-yue, Zang, Ming-wu, Li, He-nan, Guo, Wen-ping, Li, Ying-ying, Wang, Shou-wei
• Format: Article
• Language: English
• Subjects: Acetonitrile; Aminoglycosides; Antibiotics; Assaying; Bioassays; Biochemistry; Biological activity; Biotechnology; Boric acid; Chemistry; Chemistry and Materials Science; Correlation analysis; Crosslinking; Detection limits; E coli; Embedding; Food safety; Hexanes; Meat; Microbial activity; Microorganisms; n-Hexane; Optical density; pH effects; Polyvinyl alcohol; Quinolones; Residues; Screening; Shelf life; Sodium tetraborate; Testing laboratories; Tetracyclines; β-Lactam antibiotics
• ISSN: 0273-2289; 1559-0291
• DOI: 10.1007/s12010-020-03462-z

A preliminary screening assay based on a microbial chromogenic reaction was developed to detect common antibiotic residues in meat rapidly. The assay comprised two bioassays: one for Escherichia coli and another for Geobacillus stearothermophilus . The assay was optimized and evaluated for the simultaneous screening of 30 antibiotics from five common antibiotic classes (tetracyclines, aminoglycosides, macrolides, β -lactams, and quinolones) found in meat. Extraction using phosphate-acetonitrile buffer (pH 7.2) and a delipidating treatment using n-hexane resulted in a high extraction efficacy for the five antibiotics, without affecting the microbial color reaction. A carrier, polyvinyl alcohol (0.1 g/mL); a cross-linking agent, boric acid-sodium tetraborate solution (pH 5.5); and a bacterial suspension with an initial optical density of 1.0 were the optimal embedding conditions for stability, microbial activity, and chromogenic efficiency. The assay exhibited a 6-month shelf life, with detection limits of 40–60, 60–140, 60–100, 20–40, and 40–180 μg/kg for tetracyclines, aminoglycosides, macrolides, β -lactams, and quinolones, respectively, which met the European Commission (37/2010) requirements for antibiotic residue limits. Our assay results were confirmed using LC-MS/MS with 160 samples, revealing a good correlation. This study demonstrates a reliable, easy-to-use, and economical method for preliminary screening of antibiotic residues in meat. This method may find an immediate application in food safety and general testing laboratories.