College of Food Science, Southwest University, No.2 Tiansheng Road, Beibei District, Chongqing, 400715, China.
Biological Science Research Center, Southwest University, No.2 Tiansheng Road, Beibei District, Chongqing, 400715, China.
Anal Bioanal Chem. 2020 Sep;412(23):5627-5635. doi: 10.1007/s00216-020-02778-3. Epub 2020 Jun 30.
Zearalenone (ZEN) is a type of estrogenic mycotoxin commonly occurring in cereals. The aim of this study was to design a simple, rapid, inexpensive and ultrasensitive fluorescence assay for the determination of ZEN. Here, amino-modified mesoporous silica nanoparticles (MSNs-NH) were synthesized to be the positive charge-rich reactor. A 6-carboxy-fluorescein-labeled aptamer (aptamer-FAM) was designed as the signal probe, ZEN-capture probe and negative charge reactor. In the absence of ZEN, the negatively charged aptamer-FAM combined with the positively charged MSNs-NH in an electrostatic manner. In the presence of ZEN, the fluorescence intensity in the supernatant increased significantly because the aptamer-FAM could bind to ZEN instead of MSNs-NH. Under the optimal experimental conditions, this assay exhibited excellent specificity, repeatability and a wide linearity range of 0.005-150 ng/mL, with a detection limit of 0.012 ng/mL. Additionally, it showed high recovery (83.3-101.5%) for the spiked samples. There was no statistically significant difference in the ZEN concentrations detected by the proposed assay and HPLC in naturally contaminated samples. Overall, this design provides a new strategy for the rapid, inexpensive and sensitive detection of ZEN, and it could be applied to develop fluorometric assays for different targets by the selection of appropriate aptamers. Graphical abstract.
玉米赤霉烯酮(ZEN)是一种常见于谷物中的雌激素真菌毒素。本研究旨在设计一种简单、快速、廉价且超灵敏的荧光测定法来测定 ZEN。在这里,合成了氨基修饰的介孔硅纳米粒子(MSNs-NH)作为带正电荷的富反应剂。设计了一种 6-羧基-荧光素标记的适体(aptamer-FAM)作为信号探针、ZEN 捕获探针和带负电荷的反应剂。在没有 ZEN 的情况下,带负电荷的 aptamer-FAM 以静电方式与带正电荷的 MSNs-NH 结合。在存在 ZEN 的情况下,由于 aptamer-FAM 可以与 ZEN 结合而不是与 MSNs-NH 结合,因此上清液中的荧光强度显著增加。在最佳实验条件下,该测定法表现出优异的特异性、重复性和 0.005-150 ng/mL 的宽线性范围,检测限为 0.012 ng/mL。此外,它对添加的样品具有高回收率(83.3-101.5%)。在天然污染样品中,所提出的测定法和 HPLC 检测到的 ZEN 浓度没有统计学上的显著差异。总体而言,该设计为快速、廉价和灵敏检测 ZEN 提供了一种新策略,并且可以通过选择合适的适体来应用于开发用于不同靶标的荧光测定法。