筛查食品中违法添加奥沙普嗪荧光免疫层析方法研究

Development of an Immunochromatographic Assay for Screening of Oxaprozin as Illegal Adulterant in Foods

目的本研究建立了一种筛查食品中违法添加的奥沙普嗪胶体金免疫检测试纸条(ICA);方法以改良活泼酯法将奥沙普嗪发表连接到BSA和OVA,合成免疫抗原和检测抗原,免疫家兔获得针对抗奥沙普嗪抗体;以胶金垫、NC膜、样品垫和吸水垫组装IAC试剂条,以标记抗体的40nm胶体金,分别与NC膜上的检测抗原和山羊抗兔IgG二抗进行免疫反应,形成检测线(T线)和质控线(C线)。结果在ELISA体系检测奥沙普嗪的IC_(50)值为13.2ng·mL^(-1),与8种相关药物交叉反应率都小于0.1%,优化IAC试剂条对奥沙普嗪的最低检测限为2.0ng·mL^(-1)。IAC试剂对30个市售样品的检测结果,与HPLC验证结果吻合。结论本方法灵敏、特异、简便、快速,是对违法添加奥沙普嗪的食品进行筛查的有效工具。

OBJECTIVE An immunochromatographic assay（ ICA） for screening oxaprozin as illicit adulterant in food was developed in this research. METHODS Artificial antigens were prepared by conjugating oxaprozin to OVA and BSA using modified active ester mechanism. Antibodies against oxaprozin were raised from rabbits by immunizing with BSA-oxaprozin conjugate. RESULTS Results indicated that the IC_（50） value for oxaprozin was 13. 2 ng·mL^（-1） in the ELISA. The cross-reactivities for 8 associate drugs were lower than 0. 1%. A consequent ICA showed detection limit of 2. 0 ng·mL^（-1） for oxaprozin. The current ICA showed good agreements with HPLC in detecting oxaprozin in 30 food samples from local market. CONCLUSION The ICA is suitable for screening illegal adulterant of oxaprozin in foodstuffs due to highly sensitive and specific.