Development of a sensitive indirect competitive enzyme-linked immunosorbent assay for high-throughput detection and risk assessment of aflatoxin B1 in animal-derived medicines
Development of a sensitive indirect competitive enzyme-linked immunosorbent assay for high-throughput detection and risk assessment of aflatoxin B1 in animal-derived medicines
复制标题
开发灵敏的间接竞争酶联免疫吸附测定法，用于动物源性药物中黄曲霉毒素 B1 的高通量检测和风险评估
DOI：
10.1016/j.toxicon.2021.04.009
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发表时间：
2021-04-27
期刊：
影响因子：
2.8
通讯作者：
Yang, Meihua
中科院分区：
文献类型：
--
作者：
Wang, Changjian;Zhang, Lei;Yang, Meihua
动物源性医学是中医学的重要组成部分。研究表明，许多动物源性药品容易受到黄曲霉毒素的污染，然而，动物源性药品的快速检测却容易被忽视。为此，我们建立了一种灵敏的间接竞争酶联免疫吸附试验(IcEL ISA)，用于快速筛选金龟子、蟑螂、家蚕和蚯蚓中的黄曲霉毒素B-1(AFB(1))。IcEL ISA方法的灵敏度显著提高。四种动物源性药材的IC_(50)范围为0.092~0.135 ng mL(-1)，检出限为0.008~0.020 ng mL(-1)。为了获得较高的准确度，对提取液和提取时间进行了考察。用该方法对138份样品进行了检测，金龟子和蚯蚓样品中AFB(1)的检出率分别为26.6%和16.7%。用液-质联用法对分析结果进行了验证，两个数据集之间具有良好的相关性，R-2值为0.999。结果表明，该方法可用于动物源性药物中黄曲霉毒素B(1)的快速检测。在此基础上，对金龟子和蚯蚓的致癌风险进行了定量风险评估，结果表明金龟子摄入AFB(1)对致癌风险有一定的威胁。
Animal-derived medicine is an important part of traditional Chinese medicine (TCM). Studies have shown that many animal-derived medicinal products are susceptible to contaminate of aflatoxins, nevertheless, the rapid detection for animal-derived medicine is prone to be ignored. Here we developed a sensitive indirect competitive enzyme-linked immunosorbent assay (icELISA) for rapid screening of aflatoxin B-1 (AFB(1)) in ground beetle, cockroach, silkworm and earthworm. The sensitivity of the icELISA method was significantly enhanced. The IC50 for the four animal-derived medicinal samples ranged from 0.092 to 0.135 ng mL(-1); the limit of detection (LOD) was 0.008-0.020 ng mL(-1). To obtain high accuracy, the extraction solution and time were evaluated. By using this method, a total of 138 samples were investigated, and the detection rates of AFB(1) in ground beetle and earthworm samples were 26.6% and 16.7%, respectively. The result was validated by liquid chromatography combined with tandem mass spectrometry, and an excellent correlation was observed between the two datasets, with a R-2 value of 0.999. Our results indicate that the proposed method can be used for the rapid detection of AFB(1) in animal-derived medicine. Furthermore, the quantitative risk assessment was conducted for ground beetle and earthworm based on the results, demonstrating that the intake of AFB(1) in ground beetle had a slight threat to the risk of cancer.