保健食品中违法添加双氯芬酸钠免疫层析筛查方法的建立

Immunochromatographic assay for the screen of illegal adulterant diclofenac sodium in health foods

目的为了筛查食品中违法添加的双氯芬酸钠,本研究建立相关胶体金免疫层析检测方法(ICA)。方法以活泼酯机制将双氯芬酸钠连接到不同蛋白质载体,分别合成免疫抗原和检测抗原,免疫家兔获得针对双氯芬酸钠的特异性抗体;抗体标记的胶体金(40 nm)包被在胶体金垫,检测抗原和二抗喷涂在硝酸纤维素膜(NC膜)上,分别构建检测线(T线)和质控线(C线)。组装胶金垫、NC膜、样品垫和吸水垫,组成ICA试剂条。结果在ELISA体系检测双氯芬酸钠的IC50值为9.3 ng·mL^(-1),与8种相关药物交叉反应率都小于0.01%,以ICA试剂条对双氯芬酸钠的最低检测限为2.0 ng·mL^(-1)。IAC试剂对60个市售样品的检测结果,HPLC验证完全吻合。结论本方法简便、价廉、灵敏、特异,适用于筛查食品中违法添加的双氯芬酸钠。

Objective To develop an immunochromatographic assay （ICA） to screen illegal adulterant diclofenac sodium in health food. Methods Diclofenac sodium was conjugated to different protein carriers to synthesize artificial antigens by active ester mechanism. Specific antibodies were obtained from rabbits immunized by immunogen. Results The ICso value for diclofenac sodium was 9.3 ng mL-1 by ELISA. The cross- reaction rates of 8 related drugs were lower than 0.01%. The consequent ICA showed the detection limit of 2.0 ng mL-1 for diclofenac sodium. Good agreements were observed between current ICA and HPLC in detecting diclofenac sodium in 60 batches of health food samples. Conclusion ICA is a good tool to screen diclofenac sodium as illegal adulterant because it is highly sensitive and specific.