Microchimica Acta

, 186:225 | Cite as

Immunochromatographic fluorometric determination of clenbuterol with enhanced sensitivity

  • Yuyang Zeng
  • Demei Liang
  • Pimiao Zheng
  • Tao Peng
  • Shujuan Sun
  • Ghulam Mujtaba Mari
  • Haiyang JiangEmail author
Original Paper


A method is described to enhance the sensitivity of an immunochromatographic assay for clenbuterol (CLE) by making use of dually-labeled gold nanoparticles (GNPs), background fluorescence blocking, and immunomagnetic separation. The GNPs were labeled with biotinylated antibody and streptavidin, respectively, and dually labeled GNPs were obtained via the biotin-streptavidin interaction to amplify the detection signal. The fluorescent signal was blocked by dually labeled GNPs and decreased as the dually labeled GNPs aggregation increases on nitrocellulose membrane, which derived from fluorescent polyvinylchloride card. However, fluorescence (measured at excitation/emission wavelengths of 518/580 nm) recovers when CLE reacts with dually labeled GNPs. Immunomagnetic separation was first applied for sample pretreatment. This can offset the matrix effect and improves the sensitivity and accuracy of the assay. Under the optimal conditions, the limits of detection of CLE visually were 0.25 μg·L−1. In addition, clenbuterol can be quantified in swine urine with a 0.03 μg·L−1 detection limit. This is 60-fold lower than current immunochromatography. Response is linear in the 0.06–0.59 μg·L−1 concentration range, and the recoveries from spiked swine urine range from 81 to 115%.”

Graphical abstract

Schematic presentation of the strategies for improving sensitivity of immunochromatographic assay. It includes immunomagnetic separations, dually-labeled gold nanoparticles and background fluorescence blocking. The assay was applied to detect clenbuterol (CLE) in swine urine with an excellent performance.


Lateral flow immunoassay Dual-labeled gold nanoparticles Background fluorescence Fluorescence blocking Immunomagnetic separation Signal amplification Clenbuterol Swine urine 



This study was supported financially by grants from the Ministry of Science and Technology of People’s Republic of China (2017YFF0211003).

Compliance with ethical standards

The author(s) declare that they have no competing interests.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Informed consent

Not applicable.

Supplementary material

604_2019_3326_MOESM1_ESM.docx (179 kb)
ESM 1 (DOCX 179 kb)


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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2019

Authors and Affiliations

  • Yuyang Zeng
    • 1
  • Demei Liang
    • 1
  • Pimiao Zheng
    • 1
  • Tao Peng
    • 1
  • Shujuan Sun
    • 1
  • Ghulam Mujtaba Mari
    • 1
  • Haiyang Jiang
    • 1
    Email author
  1. 1.Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Veterinary Medicine, Beijing Key Laboratory of Detection Technology for Animal-Derived Food Safety, Beijing Laboratory for Food Quality and SafetyChina Agricultural UniversityBeijingPeople’s Republic of China

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