A Novel Indirect Competitive Enzyme-Linked Immunosorbent Assay Format for the Simultaneous Determination of Ractopamine and Phenylethanolamine A Residues in Swine Urine
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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.
KeywordsRactopamine Phenylethanolamine A Indirect competitive enzyme-linked immunosorbent assay Monoclonal antibody Residues
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.
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.
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