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目的优化硝苯地平药物的免疫胶体金探针的制备条件。方法采用柠檬酸三钠还原法制备胶体金溶液,采用紫外分光光度法在400~700 nm处测定胶体金溶液吸光度,判断胶体金颗粒大小。调整标记胶体金溶液p H、抗体蛋白量以及复溶液等因素,试制试纸条并进行加标验证实验,确定制备免疫胶体金探针的最优条件。结果实验制备胶体金颗粒大小在20~40 nm;最佳制备条件为:单抗标记p H为9.0,单抗标记量为5.0μL,复溶液为0.5 mol/L Tris(p H 9.0,含0.5%Tween20)。经加标实验验证,试纸条的检测灵敏度可以达到设计要求。结论硝苯地平免疫胶体金探针的制备条件优化,可为下一步制作高质量的免疫胶体金试纸条提供技术准备,可应用于对食品中非法添加硝苯地平的快速检测。
Objective To optimize the preparation conditions of immunogold gold probe of nifedipine. Methods Colloidal gold solution was prepared by sodium citrate reduction method. The absorbance of colloidal gold solution was measured by ultraviolet spectrophotometry at 400-700 nm to determine the colloidal gold particle size. Adjust the tagged gold solution p H, the amount of antibody protein and complex solution and other factors, the test strips and spike validation experiments to determine the optimal conditions for the preparation of immunogold gold probes. Results The size of colloidal gold particles prepared in the experiment was 20-40 nm. The best preparation conditions were as follows: the mAb labeled p H was 9.0, the labeled amount of the mAb 5.0 μL and the reconstituted solution 0.5 mol / L Tris (p H 9.0, containing 0.5 % Tween20). After the standard test verification, the test strip test sensitivity can meet the design requirements. Conclusion The optimization of the preparation conditions of nifedipine immunogold probe can provide the technical preparation for the next step of producing high quality immunocolloid gold test strip, which can be applied to the rapid detection of illegally added nifedipine in food.