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An oligosorbent-based aptamer affinity column for selective extraction of aflatoxin B2 prior to HPLC with fluorometric detection

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Abstract

The article describes an aptamer affinity column for selective solid-phase extraction of aflatoxin B2 (AFB2). Amino-modified aptamer against AFB2 was immobilized on CNBr-activated Sepharose through a covalent bond. The effects of oligosorbents based on 3′- or 5′-amino-modified sequences with a C6 or a C7 spacer arm were evaluated by UV spectroscopy at 260 nm. The extraction recovery was evaluated by HPLC with fluorometric detection. The extraction of AFB2 was optimized. Under the optimum conditions, the aptamer affinity column has a linear response to AFB2 in the range of 0.5–80 ng, with a capacity of 84.6 ng. Control supports without immobilized aptamers and a nonspecific oligosorbent immobilized with a negative control oligonucleotide were studied in order to demonstrate selectivity. The method was tested with spiked peanut sample (0.5–50 μg·kg−1 AFB2) and gave average recoveries of 80.9% and a mean relative standard deviation of 1.9%. The limit of detection is 25 pg·mL−1. This is much lower than the maximum residue limits suggested by the European Union. The columns can be re-used up to five times without any loss of performance. The oligosorbent was also applied to clean-up of AFB2 from peanut sample extracts before HPLC analysis. Results were further confirmed by ultra-fast liquid chromatography with tandem mass spectrometry. Conceivably, the method may also be applied to other samples, such as food, agricultural products, and traditional Chinese medicines.

Schematic illustration of the fabrication procedures of aptamer affinity column, AAC (a), its principle of aptamer bound to aflatoxin B2 (b) and the obtained AAC (c).

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Acknowledgements

The authors are grateful for the support from the Science and technology innovation special construction funded program of Bei**g academy of agriculture and forestry sciences (KJCX20170401 and KJCX20170419), the Science and technology project of Bei**g (Z171100001317012), the Chinese Ministry of Agriculture and the Open Project of Bei**g Research Center for Agricultural Standards and Testing (ATFM-KFKT201601), and the Chinese postdoctoral science foundation funded project (2017 M610807).

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Authors and Affiliations

  1. Bei**g Research Center for Agricultural Standards and Testing, Agricultural Product Quality and Safety Risk Assessment Laboratory of the Department of Agriculture, Bei**g Municipal Key Laboratory of Agriculture Environment Monitoring, Bei**g, 100097, China

    Hongmei Liu, Yunxia Luan, Anxiang Lu, Bingru Li & JiHua Wang

  2. Key Laboratory of Bioactive Substances and Resources Utilization of Chinese Herbal Medicine, Ministry of Education, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Bei**g, 100193, China

    Meihua Yang

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  1. Hongmei Liu
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Liu, H., Luan, Y., Lu, A. et al. An oligosorbent-based aptamer affinity column for selective extraction of aflatoxin B2 prior to HPLC with fluorometric detection. Microchim Acta 185, 71 (2018). https://doi.org/10.1007/s00604-017-2591-7

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