Development of a Competitive Indirect Enzyme-Linked Immunosorbent Assay for Screening Phenylethanolamine A Residues in P
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Development of a Competitive Indirect Enzyme-Linked Immunosorbent Assay for Screening Phenylethanolamine A Residues in Pork Samples Xiaomei Wang 1 & Tongling Liufu 1 & Natalia V. Beloglazova 2 & Pengjie Luo 3 & Jianwen Qu 1 & Wenxiao Jiang 1
Received: 18 November 2015 / Accepted: 28 March 2016 # Springer Science+Business Media New York 2016
Abstract Phenylethanolamine A (PEA), a new alternative βagonist, has been illegally used in farming to promote the muscle growth in food-producing animals. In this study, a sensitive and convenient competitive indirect enzyme-linked immunosorbent assay (ciELISA) was developed for determination of PEA residues in pork samples. The produced antibody was highly specific to PEA and exhibited a negligible cross-reactivity toward some other β-agonists. The developed technique was characterized by the limit of detection below 0.08 μg kg −1 and the IC 50 value of 0.93 pmol mL −1 (0.32 ng mL−1). Validation of the technique was done using artificially spiked and naturally contaminated pork samples. The recoveries ranged from 79.6 to 112.6 % for the samples spiked at levels of 0.1–5 μg kg−1 with the variation coefficients below 15 %. The analysis of naturally contaminated samples showed that the obtained data corresponded with the data obtained by the LC-MS/MS. The developed ciELISA was shown to be a feasible highly sensitive and specific screening tool for PEA residue analysis.
* Wenxiao Jiang [email protected]
1
Key Laboratory of Biomedical Engineering, School of Medicine, Health Science Center, Shenzhen University, Shenzhen 518060, China
2
Laboratory of Food Analysis, Department of Bioanalysis, Faculty of Pharmaceutical Sciences, Ghent University, Ottergemsesteenweg 460, Ghent B-9000, Belgium
3
Key Laboratory of Food Safety Risk Assessment, Ministry of Health, China National Center for Food Safety Risk Assessment, Beijing 100021, China
Keywords Phenylethanolamine A . Antibody . Pork . Residue . Food safety . ciELISA
Introduction β-agonists, a class of sympathomimetic agents acting upon βadrenoceptors, have been originally used for preventing preterm labor and treatment of asthma and other respiratory diseases in humans (Brambilla et al. 2000; Rosales-Conrado et al. 2015). Besides, β-agonists are also misused as growth promoters in livestock production, because these compounds can increase feed efficiency and enhance lean-to-fat ratio (Mitchell and Dunnavan 1998). The most commonly abused β-agonists are clenbuterol, salbutamol, and ractopamine. However, a long-term or high-dose use of β-agonists can lead to the deleterious physiological side effects and trigger the acute toxic responses such as cardiac palpitation, tachycardia, nervousness, muscle tremors, and confusion (Fang et al. 2011; Tang et al. 2015). Besides, residues of these chemicals can pose the potential human health hazards, such as food poisoning, cardiovascular, or central nervous diseases (Malucelli et al. 1994). Therefore, the Ministry of Agriculture of China and some other countries have banned β-agoni
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