中药材中黄曲霉毒素B_1免疫学快速检测方法研究

Study on rapid immunological detection method for determining AFB_1 in traditional Chinese medicines

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.