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[中药黄曲霉毒素酶联免疫吸附测定法的建立]

[Development of enzyme linked immunosorbent assay of aflatoxin of Chinese herbal medicines].

作者信息

Tie-Gui Nan, Xiao-Xu Hong, Xin-Yi X U, Lu-Qi Huang, Yuan Yuan

机构信息

State Key Laboratory Breeding Base of Dao-Di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences Beijng 100700, China.

Chinese Pharmacopoeia Commission Beijing 100061, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2020 Sep;45(17):4158-4162. doi: 10.19540/j.cnki.cjcmm.20200608.101.

Abstract

The enzyme linked immunosorbent assay of aflatoxin has been adopted in Chinese Pharmacopoeia(2020 edition). Based on high-throughput screening of monoclonal antibodies technology, monoclonal antibodies that can specifically recognize the aflatoxin B_1 and the total amount of aflatoxin B_1, B_2, G_1, and G_2 in Chinese herbal medicines were prepared. By optimizing the concentration of coating antibody, enzyme-labeled antigen, and the reaction system of enzyme-linked immunosorbent assay, the enzyme linked immunosorbent assay(ELISA) were developed for detection of aflatoxins in Chinese herbal medicines, decoction pieces, and preparation of Chinese medicine. In this method, the recovery test of actual samples is 60%-120%, and the relative standard deviation is less than 15%. In addition, in view of the complicated and expensive pretreatment methods for the determination of aflatoxin in Chinese herbal medicine, we developed a highly efficient pretreatment method of liquid-liquid extraction of aflatoxin in Chinese herbal medicine without immunoaffinity column. As an effective method for the detection of aflatoxin, the ELISA can effectively reduce the aflatoxins testing cost of traditional Chinese medicine, and promote the detection ability at earlier stages of production, and strengthen the quality supervision of traditional Chinese medicine.

摘要

黄曲霉毒素的酶联免疫吸附测定法已被《中国药典》(2020年版)采用。基于高通量筛选单克隆抗体技术,制备了能够特异性识别中药材中黄曲霉毒素B_1以及黄曲霉毒素B_1、B_2、G_1和G_2总量的单克隆抗体。通过优化包被抗体、酶标抗原的浓度以及酶联免疫吸附测定的反应体系,建立了用于检测中药材、中药饮片和中成药中黄曲霉毒素的酶联免疫吸附测定法(ELISA)。该方法实际样品回收率为60%-120%,相对标准偏差小于15%。此外,鉴于中药材中黄曲霉毒素测定的预处理方法复杂且昂贵,我们开发了一种无需免疫亲和柱的中药材中黄曲霉毒素液液萃取高效预处理方法。作为一种有效的黄曲霉毒素检测方法,ELISA可有效降低中药黄曲霉毒素检测成本,提升生产前期检测能力,加强中药质量监管。

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