期刊
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
卷 412, 期 4, 页码 895-904出版社
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-019-02300-4
关键词
Aptamer; Aflatoxin B-1; Aflatoxin B-2; Aptamer affinity column; Agro-products
资金
- Beijing Municipal Natural Science Foundation [7192026] Funding Source: Medline
- National Natural Science Foundation of China [81803712] Funding Source: Medline
- Beijing academy of agriculture and forestry sciences [KJCX20170401] Funding Source: Medline
- Beijing agricultural forestry academy youth foundation [QNJJ201630] Funding Source: Medline
- Beijing Excellent Talent Training Fund [2017000020060G131] Funding Source: Medline
We have developed an aptamer affinity column (AAC) for the purification and enrichment of trace aflatoxin B-1 (AFB(1)) and aflatoxin B-2 (AFB(2)) 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 AFB(1) and 162.5 +/- 8.9 ng for AFB(2)), selectivity (excellent), reusability (twenty-three times for AFB(1) and twelve times for AFB(2)), and repeatability (92.7% +/- 2.9% for AFB(1) and 71.5% +/- 3.4% for AFB(2)). 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 AFB(1) and 15 pg mL(-1) for AFB(2), and acceptable recovery with different spiking levels in different matrices. Finally, the AAC was successfully applied to analyte AFB(1) and AFB(2) 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. An aptamer affinity column for purification and enrichment of aflatoxin B-1 and aflatoxin B-2 in agro-products with the aid of HPLC-FLD and a post-column photochemical derivatization reactor
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