An aptamer affinity column for purification and enrichment of aflatoxin B1 and aflatoxin B2 in agro-products
An aptamer affinity column for purification and enrichment of aflatoxin B1 and aflatoxin B2 in agro-products
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适配体亲和柱纯化富集农产品中黄曲霉毒素B1和黄曲霉毒素B2
DOI：
10.1007/s00216-019-02300-4
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发表时间：
2020-02-01
影响因子：
4.3
通讯作者：
Luan, Yunxia
中科院分区：
文献类型：
--
作者：
Liu, Hongmei;Zhao, Ying;Luan, Yunxia
我们研制了一种适配子亲和柱(AAC)，通过氨基修饰的适配子与NHS活化的琼脂糖共价连接，用于纯化和浓缩真正的农产品中的痕量黄曲霉毒素B1(AFB1)和黄曲霉毒素B2(AFB2)。考察了适合于该AFB-AAC的耦合时间(2分钟)、负载体积(30毫升)和负载步骤中使用的甲醇浓度(<； 10%)。然后从容量(黄曲霉毒素1为329.1 ± 13.7 ng和黄曲霉毒素2为162.5 ± 8.9 ng)、选择性(优良)、可重用性(黄曲霉毒素1为23倍和黄曲霉毒素2为12倍)和重复性(黄曲霉毒素1为92.7% ± 2.9%和黄曲霉毒素2为71.5% ± 3.4%)等方面进一步评估了黄曲霉毒素B-AAC的性能。此外，AAc净化与高效液相色谱联用具有良好的宽范围线性关系，对黄曲霉毒素1和黄曲霉毒素2的检出限分别为50pgmL−1和15pgmL−1，在不同基质中的加标回收率均可接受。最后，将该方法成功地应用于四种农产品和一种玉米粉标准物质中的AFB1和AFB2的分析，结果与商业IAC的结果一致。图形摘要利用高效液相色谱-荧光检测器和柱后光化学衍生化反应器纯化和浓缩农产品中黄曲霉毒素B1和黄曲霉毒素B2的适体亲和柱，借助高效液相-荧光检测器和柱后光化学衍生反应器纯化和浓缩农产品中黄曲霉毒素B1和黄曲霉毒素B2的适配子亲和色谱柱
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. Graphical abstract An aptamer affinity column for purification and enrichment of aflatoxin B1 and aflatoxin B2 in agro-products with the aid of HPLC-FLD and a post-column photochemical derivatization reactor An aptamer affinity column for purification and enrichment of aflatoxin B1 and aflatoxin B2 in agro-products with the aid of HPLC-FLD and a post-column photochemical derivatization reactor