Laboratory of Cultivation and Breeding of Medicinal Plants, National Administration of Traditional Chinese Medicine, Department of Traditional Chinese Medicine, Jilin Agricultural University, Changchun, 130118, China; Key Laboratory of Bioactive Substances and Resources Utilization of Chinese Herbal Medicine, Ministry of Education, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100193, China.
Key Laboratory of Bioactive Substances and Resources Utilization of Chinese Herbal Medicine, Ministry of Education, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100193, China; Medical Laboratory of the Third affiliated hospital of Shenzhen university, Shenzhen, 518001, China.
J Pharm Biomed Anal. 2021 Feb 20;195:113868. doi: 10.1016/j.jpba.2020.113868. Epub 2020 Dec 24.
As a highly toxic and widely used herbicide, atrazine poses a serious threat to food safety as well as overall environmental and human health. Due to complex matrix interference and the difficulty of signal enrichment, there is an urgent need for a convenient, fast, and ultrasensitive method that detects trace atrazine without concern for matrix effects. Here, we provide the first account of a sensitive and rapid suspension probe based on magnetic microspheres used to detect atrazine in herbs. The self-made magnetic beads featured -COOH groups and were used as the carrier to construct immunofluorescent probes. These probes then conjugated with the atrazine antigen through an activated ester method, ultimately binding to the antibody. Homogeneous detection was ensured using flow cytometry and the microflow optical channel along with allophycocyanin-conjugated goat-anti-mouse secondary antibody (APC-IgG-SecAb) as the fluorescent signal. The magnetic suspension probe allowed for high target enrichment and the inherent two-dimensional selective detection of flow cytometry effectively avoided any matrix interference. This method had good linearity across 1.69-23.19 ng mL. The IC and LOD values were 4.81 ng mL and 0.95 ng mL, respectively; the sensitivity was increased three-fold relative to ELISA. After complete optimization, 2-N-morpholinoeth-anesulfonic acid was used as the coupling solution and maintained good mono-dispersity, stability, and reactivity for the labelled microspheres during the process. The entire experiment was simple, and effectively used reagents; moreover, both the labor required and detection time were greatly reduced. Critically, the strategy presented here greatly reduced interference from complex matrices, and saved preparation for matrix-matched solutions when different herbs were screened. Overall, this strategy was sensitive, rapid, eco-friendly, and labor-saving; collectively, these attributes make it well-suited for on-site screening of atrazine contamination and will allow for increased food safety.
作为一种高毒性且广泛使用的除草剂，阿特拉津对食品安全以及整体环境和人类健康构成严重威胁。由于复杂基质干扰和信号富集的困难，迫切需要一种方便、快速和超灵敏的方法，能够检测痕量阿特拉津而无需担心基质效应。在这里，我们首次提供了一种基于磁性微球的灵敏快速悬浮探针用于检测草药中阿特拉津的方法。自制的磁性微球具有-COOH 基团，用作载体构建免疫荧光探针。这些探针通过活化酯法与阿特拉津抗原偶联，最终与抗体结合。通过使用流式细胞术和微流光学通道以及别藻蓝蛋白偶联的山羊抗小鼠二级抗体（APC-IgG-SecAb）作为荧光信号，实现了均相检测。磁性悬浮探针允许高目标富集，而流式细胞术固有的二维选择性检测有效地避免了任何基质干扰。该方法在 1.69-23.19 ng mL 范围内具有良好的线性。IC 和 LOD 值分别为 4.81 ng mL 和 0.95 ng mL，灵敏度比 ELISA 提高了三倍。经过完全优化，使用 2-N-吗啉乙磺酸作为偶联溶液，在标记微球过程中保持良好的单分散性、稳定性和反应性。整个实验简单，有效使用试剂，大大减少了所需的劳动力和检测时间。重要的是，这里提出的策略大大减少了复杂基质的干扰，并且在筛选不同草药时节省了基质匹配溶液的制备。总的来说，该策略具有灵敏度高、快速、环保和省力的特点，非常适合现场筛选阿特拉津污染，将提高食品安全水平。
