Advertisement

A Novel Indirect Competitive Enzyme-Linked Immunosorbent Assay Format for the Simultaneous Determination of Ractopamine and Phenylethanolamine A Residues in Swine Urine

  • Dapeng Peng
  • Lan Zhao
  • Lingyan Zhang
  • Yuanhu Pan
  • Yanfei Tao
  • Yulian WangEmail author
  • Feng ShengEmail author
  • Zonghui YuanEmail author
Article
  • 14 Downloads

Abstract

For monitoring ractopamine (RAC) and phenylethanolamine A (PA) with high efficiency, a novel indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) format was developed. At first, two specific monoclonal antibodies (mAbs) against RAC and PA were prepared, respectively. Based on the finding that both mAbs RAC and PA could recognize the same coating antigen RAC-SA-OVA, a novel ic-ELISA format was constructed, in which the limit of detection (LOD), recoveries, and coefficient of variation of the ic-ELISA developed for RAC residues were 0.35 μg L−1, 96.7~108.6%, and less than 6.8%, and for PA residues, they were 0.11 μg L−1, 97.8~110.2%, and less than 9.4%, respectively. The developed method also exhibited a positive correlation with the results of HPLC-MS conducted on the samples. These data indicate that the developed novel ic-ELISA is reliable and could be used in a routine application for the simultaneous determination of RAC and PA residues in swine urine.

Keywords

Ractopamine Phenylethanolamine A Indirect competitive enzyme-linked immunosorbent assay Monoclonal antibody Residues 

Notes

Funding

The authors are grateful to the National Natural Science Foundation of China (31772074) and the Fundamental Research Funds for the Central Universities (2662017PY049, 2017BC010) for their financial support.

Compliance with Ethical Standards

Conflict of Interest

Dapeng Peng declares that he has no conflict of interest. Lan Zhao declares that she has no conflict of interest. Lingyan Zhang declares that she has no conflict of interest. Yuanhu Pan declares that he has no conflict of interest. Yanfei Tao declares that she has no conflict of interest. Yulian Wang declares that she has no conflict of interest. Feng Sheng declares that he has no conflict of interest. Zonghui Yuan declares that he has no conflict of interest.

Ethical Approval

This article does not contain any studies with human subjects. All animal experiments described in the present study were performed in adherence to Huazhong Agricultural University animal experiment center guidelines and approved by Animal Ethics Committee.

Informed Consent

Not applicable.

References

  1. Bai Y, Liu Z, Bi Y, Wang X, Jin Y, Sun L, Wang H, Zhang C, Xu S (2012) Preparation of polyclonal antibodies and development of a direct competitive enzyme-linked immunosorbent assay to detect residues of phenylethanolamine A in urine samples. J Agric Food Chem 60:11618–11624CrossRefGoogle Scholar
  2. Burnett T, Rodewald J, Moran J, Turberg M, Brunelle S, Coleman M (2012) Determination of ractopamine in swine, bovine, and turkey tissues by HPLC with fluorescence detection: first action 2011.22. J AOAC Int 95(4):945–958CrossRefGoogle Scholar
  3. Cao B, He G, Yang H, Chang H, Li S, Deng A (2013) Development of a highly sensitive and specific enzyme-linked immunosorbent assay (ELISA) for the detection of phenylethanolamine A in tissue and feed samples and confirmed by liquid chromatography tandem mass spectrometry (LC-MS/MS). Talanta 115:624–630CrossRefGoogle Scholar
  4. Di Corcia D, Morra V, Pazzi M, Vincenti M (2010) Simultaneous determination of β2-agonists in human urine by fast-gas chromatography/mass spectrometry: method validation and clinical application. Biomed Chromatogr 24(4):358–366Google Scholar
  5. Ding G, Li D, Qin J, Zhu J, Wang B, Geng Q, Guo M, Punyapitak D, Cao Y (2015) Development and validation of a high-performance liquid chromatography method for determination of ractopamine residue in pork samples by solid phase extraction and pre-column derivatization. Meat Sci 106:55–60CrossRefGoogle Scholar
  6. Elliott C, Thompson C, Arts C, Crooks S, van Baak M, Verheij E, Andrew B (1998) Screening and confirmatory determination of ractopamine residues in calves treated with growth promoting doses of the β-agonist. Analyst 123:1103–1107CrossRefGoogle Scholar
  7. FDA (2000) New animal drugs for use in animal feeds: ractopamine hydrochloride. Fed Regist 65:4111–4112Google Scholar
  8. Gressler V, Franzen A, de Lima G, Tavernari F, Dalla Costa O, Feddern V (2016) Development of a readily applied method to quantify ractopamine residue in meat and bone meal by quechers-lc-ms/ms. J Chromatogr B:192–200–1015-1016Google Scholar
  9. Haasnoot W, Stouten P, Lommen A, Cazemier G, Hooijerink D, Schilt R (1994) Determination of fenoterol and ractopamine in urine by enzyme immunoassay. Analyst 119:2675–2680CrossRefGoogle Scholar
  10. Juan C, Igualada C, Moragues F, León N, Mañes J (2010) Development and validation of a liquid chromatography tandem mass spectrometry method for the analysis of β-agonists in animal feed and drinking water. J Chromatogr A 1217:6061–6068CrossRefGoogle Scholar
  11. Köhler G, Milstein C (1975) Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256:495–497CrossRefGoogle Scholar
  12. Li M, Chen Z, Zhu Q (2007) Development and identification of monoclonal antibodies against ractopamine. Hybridoma 26(3):148–154CrossRefGoogle Scholar
  13. Li X, Zhang G, Deng R, Yang Y, Liu Q, Xiao Z, Yang J, Xing G, Zhao D, Cai S (2010) Development of rapid immunoassays for the detection of ractopamine in swine urine. Food Addit Contam A 27(8):1096–1103CrossRefGoogle Scholar
  14. Li Y, Lu S, Liu Z, Sun L, Guo J, Hu P, Zhang J, Zhang Y, Wang Y, Ren H, Meng X, Zhou Y (2015) A monoclonal antibody based enzyme-linked immunosorbent assay for detection of phenylethanolamine A in tissue of swine. Food Chem 167:40–44CrossRefGoogle Scholar
  15. Liu L, Kuang H, Peng C, Wang L, Xu C (2013) Fragment-based hapten design and screening of a highly sensitive and specific monoclonal antibody for ractopamine. Anal Methods 6(1):229–234CrossRefGoogle Scholar
  16. Mcconnell, R., Fitzgerald, S., Benchikh, E., Lowry, A. 2007 Phenethanolamine- derived haptens, immunogens, antibodies and conjugates. US7192722Google Scholar
  17. Mcconnell, R., Fitzgerald, S., Benchikh, E., Lowry, A. 2011 Phenethanolamine- derived haptens, immunogens and conjugates comprising them and antibodies recognising said immunogenes and conjugates. EP1657235Google Scholar
  18. MOA Bulletin No. 1519. 2010 Ministry of Agriculture, PR ChinaGoogle Scholar
  19. MOA Regulation 176, 2002 Ministry of Agriculture, PR ChinaGoogle Scholar
  20. Shelver W, Smith D (2002) Application of a monoclonal antibody-based enzyme-linked immunosorbent assay for the determination of ractopamine in incurred samples from food animals. J Agric Food Chem 50:2742–2747CrossRefGoogle Scholar
  21. Shelver W, Smith D, Berry E (2000) Production and characterization of a monoclonal antibody against the β-adrenergic agonist ractopamine. J Agric Food Chem 2000(48):4020–4026Google Scholar
  22. Tao Y, Zhu F, Chen D, Xie S, Pan Y, Wang X, Liu Z, Peng D, Yuan Z (2014) Evaluation of matrix solid-phase dispersion extraction for 11 β-agonists in swine feed by liquid chromatography with electrospray ionization tandem mass spectrometry. J Sep Sci 37:2574–2582CrossRefGoogle Scholar
  23. Wang J, Wang Y, Pan Y, Chen D, Liu Z, Liang F, Peng D, Yuan Z (2017) Preparation of a generic monoclonal antibody and development of a highly sensitive indirect competitive ELISA for the detection of phenothiazines in animal feed. Food Chem 221:1004–1013CrossRefGoogle Scholar
  24. Wang J, Zhang S, Shen J (2006) Technical note: a monoclonal antibody-based immunoassay for determination of ractopamine in swine feeds. J Anim Sci 84(5):1248–1251CrossRefGoogle Scholar
  25. Wang Z, Liu M, Shi W, Li C, Zhang S, Shen J (2015) New haptens and antibodies for ractopamine. Food Chem 183:111–114CrossRefGoogle Scholar
  26. Xiong L, Gao Y, Li W, Yang X, Shimo S (2015) Simple and sensitive monitoring of β2-agonist residues in meat by liquid chromatography tandem mass spectrometry using a QuEChERS with preconcentration as the sample treatment. Meat Sci 105:96–107CrossRefGoogle Scholar
  27. Zhang M, Li C, Wu Y (2012) Determination of phenylethanolamine A in animal hair, tissues and feeds by reversed phase liquid chromatography tandem mass spectrometry with QuEChERS. J Chromatogr B 900:94–99CrossRefGoogle Scholar
  28. Zhang Y, Wang F, Fang L, Wang S, Fang G (2014) Rapid determination of ractopamine residues in edible animal products by enzyme-linked immunosorbent assay: development and investigation of matrix effects. Biomed Res Int 2009(3):269–281Google Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.National Reference Laboratory of Veterinary Drug Residues (HZAU) and MAO Key Laboratory for Detection of Veterinary Drug ResiduesHuazhong Agricultural UniversityWuhanChina
  2. 2.Hubei Collaborative Innovation Center for Green Transformation of Bio-Resources, The College of Life SciencesHubei UniversityWuhanChina

Personalised recommendations