摘要
目的建立快速、灵敏的直接竞争酶免疫分析方法测定中药材中黄曲霉素B_1(AFB_1)的含量。方法用自制AFB_1人工抗原去免疫小鼠,获取AFB_1单克隆抗体;采用碳二亚胺法将AFB_1单克隆抗体与辣根过氧化物酶偶联制备AFB_1酶标抗原,建立直接竞争酶免疫分析方法,并对检测体系的多项影响因素进行选择优化。结果该方法在1~100μg/L浓度线性关系良好(R=0.993 4),标准曲线方程为I=18.41 log C+14.54,检测限为1.98μg/L,回收率为84.6%~92.8%。结论建立的AFB_1免疫学快速检测方法操作简便、快速灵敏且选择性好,可用于中药材中AFB_1的含量测定。
Objective To establish a rapid and sensitive direct competitive enzyme-linked immunosorbent assay(ELISA) for detecting aflatoxin B_1( AFB_1) in traditional Chinese medicines. Methods The AFB_1 monoclonal antibody was obtained by self-made AFB_1 artificial antigen for immunizing mice. The AFB_1 monoclonal antibody and horseradish peroxidase(HRP) were conjugated for preparing the AFB_1enzyme-labeled antigenby by adopting the 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride(EDC) method. A direct competitive ELISA was established. Then the multiple influencing factors of detection system were selected and optimized. Results This method had good linear relation at the concentration range of 1-100 μg/L(R=0.993 4). The standard curve equation was I=18.41 log C+14.54,the detection limit was 1.98 μg/L and the recovery rate was 84.6%-92.8%. Conclusion The method established by this study is simple to operate,rapid and sensitive with good selectivity,which can be used to detect AFB_1 in traditional Chinese medicines.
作者
王彤颖
陈媛媛
高璇
余宇燕
Wang Tongying Chen Yuanyuan Gao Xuan Yu Yuyan(Pharmacy College of Fujian University of Traditional Chinese Medicine, Fuzhou , Fujian 350122, China)
出处
《现代医药卫生》
2017年第2期177-179,共3页
Journal of Modern Medicine & Health
基金
国家自然科学基金资助项目(81202914)
福建省科技厅引导性项目(2016Y0056)
福建省卫生系统中青年骨干人才培养项目(2015-ZQN-JC-33)
关键词
黄曲霉毒素B1
酶联免疫吸附测定
参考标准
酶标抗原
中药
Aflatoxin B1
Enzyme-Linked immunosorbent assay
Reference standards
Enzyme-labeled antigen
Traditional Chinese medicine