title: Rapid determination of phenylethanolamine A in biological samples by enzyme-linked immunosorbent assay and lateral-flow immunoassay.
abstract: Phenylethanolamine A (PA) is a β-adrenergic agonist, which was first used in animal husbandry as a growth promoter in China in 2010. In this study, a monoclonal-antibody (mAb)-based indirect competitive enzyme-linked immunosorbent assay (icELISA) and lateral-flow immunoassay (LFA) for the detection of PA in swine urine and pork were developed. The immunogen was prepared by linking PA hapten with carrier protein via a diazotization method. The IC50 value of the optimized icELISA was 0.44 ng mL(-1). The limits of detection of the icELISA for PA in swine urine and pork were 0.13 ng mL(-1) and 0.39 ng g(-1), respectively. The recoveries of PA from spiked swine urine and pork were in the range 82.0-107.4 % and 81.8-113.3%, respectively, with the coefficients of variation in the range 4.1-16.2% and 1.2-6.3%, respectively. The mAbs had negligible cross reactivity with 10 other β-agonists. In contrast, the LFA had a cut-off level of 5 ng mL(-1) (g) in swine urine and pork, and the results could be achieved within 5 min. Ten blind samples of swine urine were analyzed simultaneously by icELISA, LFA, and ultra-high-performance liquid chromatography-tandem mass spectrometry, and the results of the three methods agreed well. Therefore, the combination of two immunoassays provides an effective and rapid screening method for detection of PA residues in biological samples.
authors.affiliation: College of Veterinary Medicine, China Agricultural University, No. 2 Yuanmingyuan West Road, Beijing, 100193, China.|Beijing WDWK Biotechnology Company, Ltd., No. 156 Beiqing Road, Beijing, 100085, China.
authors.forename: Xiangmei
authors.initials: X
authors.lastname: Li
authors.affiliation: Beijing WDWK Biotechnology Company, Ltd., No. 156 Beiqing Road, Beijing, 100085, China.
authors.forename: Wenjun
authors.initials: W
authors.lastname: Wang
authors.affiliation: College of Veterinary Medicine, China Agricultural University, No. 2 Yuanmingyuan West Road, Beijing, 100193, China.
authors.forename: Limiao
authors.initials: L
authors.lastname: Wang
authors.affiliation: College of Veterinary Medicine, China Agricultural University, No. 2 Yuanmingyuan West Road, Beijing, 100193, China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Wang
authors.affiliation: College of Veterinary Medicine, China Agricultural University, No. 2 Yuanmingyuan West Road, Beijing, 100193, China.
authors.forename: Xingyao
authors.initials: X
authors.lastname: Pei
authors.affiliation: College of Veterinary Medicine, China Agricultural University, No. 2 Yuanmingyuan West Road, Beijing, 100193, China. wdwk001@163.com.
authors.forename: Haiyang
authors.initials: H
authors.lastname: Jiang
mesh_terms: D015078:2-Hydroxyphenethylamine / Q000031:analogs & derivatives* / Q000032:analysis / Q000276:immunology / Q000652:urine; D000318:Adrenergic beta-Agonists / Q000032:analysis* / Q000276:immunology / Q000652:urine*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation / Q000379:methods*; D004867:Equipment Design; D006133:Growth Substances / Q000032:analysis* / Q000652:urine; D057230:Limit of Detection; D051379:Mice; D011934:Reagent Strips / Q000032:analysis; D000069466:Red Meat / Q000032:analysis*; D013552:Swine / Q000652:urine*
chemical_list: C000608765:2-(4-(nitrophenyl)butan-2-ylamino)-1-(4-methoxyphenyl)ethanol; D000318:Adrenergic beta-Agonists; D000911:Antibodies, Monoclonal; D006133:Growth Substances; D011934:Reagent Strips; D015078:2-Hydroxyphenethylamine
keywords: Biological sample; Indirect competitive ELISA; Lateral-flow immunoassay; Phenylethanolamine A; β-Adrenergic agonist