An Enzyme-Linked Immunosorbent Assay to Detect Salinomycin Residues Based on Immunomagnetic Bead Clean-up
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An Enzyme-Linked Immunosorbent Assay to Detect Salinomycin Residues Based on Immunomagnetic Bead Clean-up Wenhao Tian 1 & Xiaoxiao Zhang 1 & Meirong Song 1 & Haiyang Jiang 1,2 & Shuangyang Ding 1,2 & Jianzhong Shen 1,2 & Jiancheng Li 1,2
Received: 9 December 2016 / Accepted: 10 March 2017 # Springer Science+Business Media New York 2017
Abstract Polyether antibiotics have been widely used for the treatment and prevention of coccidiosis in chicken farming. In the present study, an efficient, simple and inexpensive competitive indirect enzyme-linked immunosorbent assay (ciELISA) method based on immunomagnetic sample clean up was developed for salinomycin. Monoclonal antibodies were immobilized on the surface of carboxylic acid magnetic beads (2.8 μm in diameter). After a simple extraction, residues in sample extracts were specifically adsorbed and the supernatant removed by magnetic separation. Analytes retained on the beads were then released by elution prior to ciELISA. The limit of detection for salinomycin in chicken muscle and liver was 22 and 18 ng mL−1, respectively, and the linear quantitative range was 47–653 ng mL−1. Intra-assay recoveries ranged from 86.00 to 99.32%, and inter-assay recoveries were between 85.68 and 96.34%. The inhibition efficiency and sensitivity of this method was improved compared with traditional hydrophilic-lipophilic balance column clean up ciELISA. Furthermore, the convenience and repeatability of the immunomagnetic cleanup renders the new method of high value for the analysis of drug residues in complex matrices.
Keywords Magnetic bead . Salinomycin . Clean up . Elisa . Chicken tissue . Residue
* Jiancheng Li [email protected] 1
National Reference Laboratory of Veterinary Drug Residues, College of Veterinary Medicine, China Agricultural University, Beijing 100193, People’s Republic of China
2
Beijing Key Laboratory of Detection Technology for Animal-Derived Food Safety, Beijing Laboratory for Food Quality and Safety, Beijing 100193, People’s Republic of China
Introduction Polyether antibiotics, also known as ionophore anticoccidial drugs, are widely used in veterinary medicine and as feed additives for the prevention of coccidiosis in chickens. Polyether antibiotics have a high molecular weight and are insoluble in water but soluble in methanol, isooctane and ether. The term ‘molecular skeletons’ is used to refer to open-chain polycyclic ether structures linked by the oxygen-containing heterocyclic rings tetrahydrofuran or tetrahydropyran (Fig. 1). Alkaline metal ions can be chelated by the ether oxygen atoms of these compounds and carried through the cell membrane, which can alter the osmotic balance and cause depolarization or cell death (Stanker et al. 1994). The toxicity associated with polyether antibiotics might be due to excessive dosage, inhomogeneously mixed feeds or administration in non-target animals. Oral median lethal dose (LD50) values for monensin, salinomycin, lasalocid and maduramycin in mice were reported as 44, 50, 146 and 35 mg kg−1, respectively (Dowl
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