直接竞争荧光免疫法测定中药材中的黄曲霉毒素B_1  

Determination of Aflatoxin B_1 in Traditional Chinese Medicines by Direct Competitive Fluorescence Immunoassay

作　　者：余宇燕[1] 邱亚利[1] 张红艳[1] 邹艳辉[1] 王彤颖 

机构地区：[1]福建中医药大学药学院,福州350122

出　　处：《化学通报》2015年第9期830-834,共5页Chemistry

基　　金：国家自然科学基金项目(81202914);福建省中药学重点实验室平台资助

摘　　要：本文采用Mannich反应合成黄曲霉毒素B1（AFB1）人工抗原,免疫小鼠制备AFB1单克隆抗体。采用直接搅拌法将异硫氰酸荧光素（FITC）标记AFB1抗体,经Sephadex-50凝胶柱纯化,制得FITC-AFB1荧光标记抗体,分析其免疫学特性,从而建立了一种快速灵敏的直接竞争荧光免疫分析方法（FIA）以用于检测中药材中的AFB1含量。结果表明,AFB1-FITC标记抗体的结合比率为4.19。通过对检测体系多项影响因素的筛选优化,FIA检测方法的标准曲线方程为I=33.45 log C＋25.55,R=0.9913,线性检测范围1-100 ng/m L,检测限0.69ng/m L,回收率90.4%-106.6%。该方法具有操作简单、快速灵敏、特异性高等特点,可用于中药材中AFB1的分析测定。AFB1 artificial antigen was synthesized by Mannieh reaction, and the monoclonal antibody with high affinity and specificity was obtained after immunization of Balb/c mice. The conjugates of AFB1 antibody and FITC were prepared by direct label method. A direct competitive FIA method was established to develop a rapid and sensitive fluorescence immunoassay for the detection of AFB1. The resutts showed that the F/P ratio of labeling reaction was 4. 19. At the optimal conditions, the FIA standard curve was drawn up and the regression equation was obtained, 1 = 34.45 logC ＋ 25.55, correlation coefficient R = 0. 9913, and the range of linearity was 1 - 100 ng/mL. By FIA method, the 84.6% - 92.8% of average recoveries of AFB1 were reached. The FIA method has been successfully applied to the detection of AFBI in traditional Chinese medicines.

关 键 词：黄曲霉毒素B1 单克隆抗体 荧光标记抗体 FIA 中药材 

分 类 号：R284.1[医药卫生—中药学] O657.3[医药卫生—中医学]