We have developed an aptamer affinity column (AAC) for the purification and enrichment of trace aflatoxin B1 (AFB1) and aflatoxin B2 (AFB2) in genuine agro-products through the covalent conjugation of amino modified aptamer and NHS-activated Sepharose. The coupling and working conditions found to be suitable for this AFB-AAC were examined in regard to coupling time (2 min), loading volume (30 mL), and the methanol concentration (< 10%) used in the loading step. The performance of AFB-AAC was then further evaluated in terms of capacity (329.1 ± 13.7 ng for AFB1 and 162.5 ± 8.9 ng for AFB2), selectivity (excellent), reusability (twenty-three times for AFB1 and twelve times for AFB2), and repeatability (92.7% ± 2.9% for AFB1 and 71.5% ± 3.4% for AFB2). Furthermore, the AAC clean-up combined with HPLC-FLD demonstrated excellent linearity over a wide range, good sensitivity with an LOD of 50 pg mL−1 for AFB1 and 15 pg mL−1 for AFB2, and acceptable recovery with different spiking levels in different matrices. Finally, the AAC was successfully applied to analyte AFB1 and AFB2 in four types of agro-products as well as a maize flour reference material, and the results were found to be in accordance with those of commercial IACs. This study provides a reference for the analysis of other trace analytes by merely changing the corresponding aptamer and represents a strong contender for immune affinity columns.
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The authors are very grateful for powerful suggestions from anonymous referees. We also thank the support from the science and technology innovation special construction funded program of National Natural Science Foundation of China (81803712), Beijing Academy of Agriculture and Forestry Sciences (KJCX20170401), Beijing Agricultural Forestry Academy Youth Foundation (QNJJ201630), Beijing Municipal Natural Science Foundation (7192026), and Beijing Excellent Talent Training Fund (2017000020060G131).
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Liu, H., Zhao, Y., Lu, A. et al. An aptamer affinity column for purification and enrichment of aflatoxin B1 and aflatoxin B2 in agro-products. Anal Bioanal Chem 412, 895–904 (2020). https://doi.org/10.1007/s00216-019-02300-4
- Aflatoxin B1
- Aflatoxin B2
- Aptamer affinity column