Microchimica Acta ( IF 5.3 ) Pub Date : 2021-09-14 , DOI: 10.1007/s00604-021-04997-6 Tingting Zhao 1 , Xinyue Ding 1 , Chenchen Lin 1 , Xucong Lin 1 , Zenghong Xie 1
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A photo-initiated polymerized oligonucleotide-grafted hydrophilic affinity monolithic column was synthesized in situ, and exploited for selective in-tube solid phase micro-extraction (IT-SPME) protocol towards the sensitive detection of ochratoxin A (OTA). Only 7 min was required for the rapid polymerization of aptamer-based affinity monolith, which was much less than the reaction time of most thermal polymerization (12–16 h) and sol-gel chemistry methods (up to 52 h). Characterizations such as polymerization recipes, structure morphology, FTIR spectrum, elemental mapping, mechanical stability, and specific recognition performance were evaluated. A significantly hydrophilic nature with a low contact angle of 15° was observed, and a mixed-mode mechanism including aptamer affinity recognition and hydrophilic interaction (HI) was employed. By coupling with HPLC-fluorescence detection, the highly specific online recognition performance was achieved with an extremely low nonspecific adsorption of the analogues. The calibration curve of OTA was obtained in the concentration range 0.05–50.00 ng·mL−1 with a limit of detection (LOD, S/N = 3) of 0.012 ng·mL−1. Applied to sample analysis, acceptable recovery yields of 95.1 ± 1.4% – 99.5 ± 2.2% (n = 3) were obtained in beer and red wine. The proposed method lighted a promising way to efficiently preparing a hydrophilic aptamer-affinity monolith for highly specific recognition of trace mycotoxin by IT-SPME coupled with HPLC.
Graphical abstract
中文翻译：
原位光引发聚合寡核苷酸功能化亲水性毛细管亲和单体用于高选择性管内微萃取赭曲霉毒素 A 霉菌毒素
原位合成了一种光引发聚合寡核苷酸接枝亲水亲和整体柱，并用于选择性管内固相微萃取 (IT-SPME) 协议，以灵敏检测赭曲霉毒素 A (OTA)。基于适配体的亲和单体的快速聚合仅需 7 分钟，这比大多数热聚合（12-16 小时）和溶胶-凝胶化学方法（最长 52 小时）的反应时间要短得多。评估了聚合配方、结构形态、FTIR 光谱、元素映射、机械稳定性和特定识别性能等表征。观察到具有 15° 低接触角的显着亲水性，并采用了包括适体亲和识别和亲水相互作用 (HI) 的混合模式机制。通过与 HPLC-荧光检测相结合，实现了高度特异性的在线识别性能，同时对类似物的非特异性吸附极低。在 0.05–50.00 ng·mL 的浓度范围内得到 OTA 的校准曲线-1，检测限（LOD，S/ N = 3）为 0.012 ng·mL -1。 应用于样品分析，啤酒和红酒的可接受回收率为 95.1 ± 1.4% – 99.5 ± 2.2% ( n = 3)。所提出的方法为高效制备亲水性适配体亲和整体材料提供了一种有前景的方法，用于通过 IT-SPME 与 HPLC 联用对痕量霉菌毒素进行高度特异性识别。
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