This study described the use of a rapid and sensitive flow injection chemiluminescence immunoassay based on the specific binding of antigens and antibodies for the detection of brombuterol residues in the swine meat and feed samples. Carboxylic resin beads were used as a solid-phase carrier for supporting more coating antigens through acylamide bonds. Under the mechanism of competitive immunity, the brombuterol in solution competed with the coating antigen immobilized on the resin beads for the limited binding sites of antibody. Then, horseradish peroxidase-labeled secondary antibodies were introduced into the immunosensor to combine with polyclonal antibodies. The chemiluminescence intensity of the luminol-p-iodophenol-H2O2 system was catalyzed by horseradish peroxidase, and thus, signal amplification was achieved. Under the optimal conditions of the concentration of coating antigen, antibody and enzymatic secondary antibody, brombuterol could be detected quantitatively. The chemiluminescence intensity decreased linearly with the logarithm of the brombuterol concentration in the range of 0.001 to 300 ng mL−1. The limit of detection (3σ) and the limit of quantitation were 0.33 pg mL−1 and 1.1 pg mL−1, respectively. This method had undergone a series of test conditions and exhibited good specificity, stability, and reproducibility. The immunosensor can also detect real samples and provide prospects for the detection of other small molecule compounds, such as isoproterenol, enrofloxacin, and ofloxacin.
Chemiluminescence immunoassay Carboxylic resin beads Signal amplification Luminol-p-iodophenol-H2O2Brombuterol
This is a preview of subscription content, log in to check access.
This work was financially supported by the Science Fund from the National Natural Science Foundation of China (no.21175097, no.31772053), the Project of State Key Laboratory of Analytical Chemistry for Life Science (SKLACLS1716), the Suzhou Industry, State and Local Joint Engineering Laboratory for Novel Functional Polymeric Materials (SYG201636), and the Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (no.YX10900212).
Compliance with Ethical Standards
Conflict of Interest
The authors have read and complied with the statement of ethical standards for manuscripts submitted to Food Analytical Methods. All the authors listed have approved the submission: Xinchun Zhou declares that she has no conflict of interest. Yuting Li declares that she has no conflict of interest. Jing Shi declares that she has no conflict of interest. Kang Zhao declares that she has no conflict of interest. Anping Deng declares that he has no conflict of interest. Jianguo Li declares that he has no conflict of interest.
This article does not contain any studies with human subject. All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.
Informed consent was obtained from all individual participants included in the study.
Chung SWC, Lam C-H (2015) Development of a 15-class multiresidue method for analyzing 78 hydrophilic and hydrophobic veterinary drugs in milk, egg and meat by liquid chromatography-tandem mass spectrometry. Anal Methods-UK 7:6764–6776. https://doi.org/10.1039/c5ay01317hCrossRefGoogle Scholar
Dong T, Yangxiao K, Zhao K, Deng A, Li J (2016) Signal amplification strategy for highly sensitive detecting brombuterol with electrochemiluminescent immunoassay by using CdSe QDs as label and gold nanoparticle as substrate. Electroanal 28:1847–1855. https://doi.org/10.1002/elan.201600332CrossRefGoogle Scholar
Du H, Chu Y, Yang H, Zhao K, Li J, She P, Zhang X, Deng A (2016) Sensitive and specific detection of a new β-agonist brombuterol in tissue and feed samples by a competitive polyclonal antibody based ELISA. Anal Methods-UK 8:3578–3586. https://doi.org/10.1039/c6ay00079gCrossRefGoogle Scholar
Hu L, Dong T, Zhao K, Deng A, Li J (2017) Ultrasensitive electrochemiluminescent brombuterol immunoassay by applying a multiple signal amplification strategy based on a PAMAM-gold nanoparticle conjugate as the bioprobe and Ag@Au core shell nanoparticles as a substrate. Microchim Acta 184:3415–3423. https://doi.org/10.1007/s00604-017-2359-0CrossRefGoogle Scholar
Sun Y, Ding C, Lin Y, Sun W, Liu H, Zhu X, Dai Y, Luo C (2018) Highly selective and sensitive chemiluminescence biosensor for adenosine detection based on carbon quantum dots catalyzing luminescence released from aptamers functionalized graphene@magnetic beta-cyclodextrin polymers. Talanta 186:238–247. https://doi.org/10.1016/j.talanta.2018.04.068CrossRefGoogle Scholar
Zhao Y, Jiang D, Wu K, Yang H, Du H, Zhao K, Li J, Deng A (2016) Development of a sensitive monoclonal antibody-based ELISA for the determination of a β-adrenergic agonist brombuterol in swine meat, liver and feed samples. Anal Methods-UK 8:6941–6948. https://doi.org/10.1039/c6ay01709fCrossRefGoogle Scholar