Immunoassays for the Rapid Detection of Gentamicin and Micronomicin in Swine Muscle
Abstract:A monoclonal antibody (mAb)-based ELISA and strip test for gentamicin (GEN) and its analogue micronomicin (MIN), are reported in this study. The conjugate gentamicin-glutaraldehyde-bovine serum albumin (GEN-GDA-BSA) was used as an immunogen. The produced anti-GEN mAB exhibited high cross-reactivity with micronomicin (MIN; 131.2) and slight or negligible cross-reactivity with other aminoglycosides. Based on this mAB, an ELISA and a strip test for GEN and MIN were developed and evaluated. The ELISA showed a 50 inhibition concentration (IC50) of 0.75 ng/mL for GEN and 0.58 ng/mL for MIN. For GEN, the average recoveries at 25200 g/kg ranged from 73 to 91, with intraday CVs of 916 and interday CVs of 815. For MIN, the average recoveries ranged from 108 to 131, with intraday CVs of 1016 and interday CVs of 815. In contrast, the strip test for GEN or MIN had a detection limit of 5 ng/mL in phosphate-buffered saline and 50 g/kg in muscle (n = 24), and the results could be judged within 10 min. The detection results of incurred samples analyzed by the strip test, ELISA, and HPLC indicated that the two immunoassays correlated well with the HPLC method and could be used as convenient tools for the rapid screening of GEN and MIN residues in swine muscle.
Document Type: Research Article
Affiliations: 1: China Agricultural University, College of Veterinary Medicine, Department of Pharmacology and Toxicology, Beijing 100193, People's Republic of China and Ministry of Agriculture Feed Safety and Bio-availability Evaluation Center, College of Veterinary Medicine, Beijing 100193, People's Republic of China. 2: China Agricultural University, College of Veterinary Medicine, Department of Pharmacology and Toxicology, Beijing 100193, People's Republic of China. 3: China Agricultural University, College of Science, Department of Chemistry, Beijing 100193, People's Republic of China.
Publication date: January 1, 2010
- The Journal of AOAC INTERNATIONAL publishes refereed papers and reviews in the fields of chemical, biological and toxicological analytical chemistry for the purpose of showcasing the most precise, accurate and sensitive methods for analysis of foods, food additives, supplements and contaminants, cosmetics, drugs, toxins, hazardous substances, pesticides, feeds, fertilizers and the environment available at that point in time. The scope of the Journal includes unpublished original research describing new analytical methods, techniques and applications; improved approaches to sampling, both in the field and the laboratory; better methods of preparing samples for analysis; collaborative studies substantiating the performance of a given method; statistical techniques for evaluating data. The Journal will also publish other articles of general interest to its audience, e.g., technical communications; cautionary notes; comments on techniques, apparatus, and reagents.
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