| 余宇燕,邱亚利,张红艳,邹艳辉,王彤颖.化学通报,2015,78(9):830-834. |
| 直接竞争荧光免疫法测定中药材中的黄曲霉毒素B1 |
| Determination of Aflatoxin B1 in Traditional Chinese Medicines by Direct Competitive Fluorescence Immunoassay |
| 投稿时间：2015-01-30 修订日期：2015-03-11 |
| DOI： |
| 中文关键词: 黄曲霉毒素B1 单克隆抗体 荧光标记抗体 FIA 中药材 |
| 英文关键词:Aflatoxin B1 Monoclonal antibody Fluorescein labeled antibody FIA Traditional Chinese medicines |
| 基金项目: |
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| 中文摘要: |
| 目的 建立快速灵敏的直接竞争荧光免疫分析方法检测中药材中AFB1含量。方法 采用Mannich反应合成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,建立AFB1荧光免疫检测方法。结果 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 = 34.45 logC + 25.55,r=0.9913,线性检测范围1~100 ng.mL-1,检测限0.69 ng.mL-1,回收率在90.4~106.6%之间。结论 本实验所建立的直接竞争荧光免疫分析方法具有操作简单、快速灵敏、特异性高等特点,可用于中药材中AFB1的分析测定。 |
| 英文摘要: |
| Objective: To develop a rapid and sensitive fluorescence immunoassay for the detection of AFB1. Methods: Synthesis of AFB1 artificial antigen by Mannich reaction, 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 employed. Results: The F/P ratio of labeling reaction was 4.19. After the optimal conditional experiment, the FIA standard curve was drawn up and the regression equation was obtained, I = 34.45 logC 25.55, correlation coefficient r=0.9913, and the range of linearity was from 1 to 100 ng.mL-1. By FIA method, the recoveries of AFB1 in traditional Chinese medicines samples were range from 90.4 to 106.6%. Conclusion: The FIA method has been successfully applied to the detection of AFB1 in traditional Chinese medicines. |
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