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Aptamer based ultrasensitive determination of the β-adrenergic agonist ractopamine using PicoGreen as a fluorescent DNA probe

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Abstract

The authors describe an aptamer based assay for determination of ractopamine (RAC) by using PicoGreen (PG) as a fluorescent probe specific for dsDNA. In the absence of RAC, the aptamer forms a duplex structure with a complementary sequence that results in enhanced PG fluorescence. Upon binding to RAC, the aptamer undergoes a structural switch. This reduces the number of DNA duplexes formed and causes a reduction of fluorescence intensity of PG as measured at excitation/emission wavelengths of 480/520 nm. Under optimized conditions, the dynamic calibration plot covers the 50 pM to 50 μM concentration range, with a 50 pM detection limit. This meets the safety supervision regulations of the European Commission in terms of residue limits of RAC in food. The method displays high selectivity over other β-adrenergic agonists including clenbuterol, dopamine and salbutamol. The assay was successfully applied to samples of swine urine at spiking levels of 7.4 nM, 22.2 nM and 37 nM. Average recoveries ranged from 95 to 110%, with an RSD of <1.5%. The method is expected to represent a promising tool for simple, rapid and sensitive on-site detection of RAC in animal products.

An aptamer based fluorescent assay for determination of ractopamine was developed with a dynamic range of 50 pM to 50 μM. The average recovery from spiked urine samples ranged from 95 to 110%, with an RSD of <1.5%.

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Acknowledgments

All authors gratefully acknowledge the funding support from the Special Fund for Agro Scientific Research in the Public Interest (201203046, 201203023).

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Authors and Affiliations

  1. Institute of Quality Standards and Testing Technology for Agro-products, Key Laboratory of Agro-product Quality and Safety, Chinese Academy of Agricultural Sciences, Beijing, 100081, China

    Chao Zhu, Guilan Zhang, Yafei Huang, Jiao Yan & Ailiang Chen

  2. Key Laboratory of Agri-food Quality and Safety, Ministry of Agriculture, Beijing, 100081, China

    Chao Zhu, Guilan Zhang, Yafei Huang, Jiao Yan & Ailiang Chen

  3. College of Food Science and Technology, Hainan University, Haikou, 570228, China

    Yafei Huang & Jiao Yan

Authors
  1. Chao Zhu
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  2. Guilan Zhang
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  3. Yafei Huang
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  4. Jiao Yan
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  5. Ailiang Chen
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Corresponding author

Correspondence to Ailiang Chen.

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Zhu, C., Zhang, G., Huang, Y. et al. Aptamer based ultrasensitive determination of the β-adrenergic agonist ractopamine using PicoGreen as a fluorescent DNA probe. Microchim Acta 184, 439–444 (2017). https://doi.org/10.1007/s00604-016-2032-z

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  • DOI: https://doi.org/10.1007/s00604-016-2032-z

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