摘要
针对个别不法厂家在辅助治疗风湿性疾病保健酒中添加双氯芬酸的问题,建立快速检测违法添加剂双氯芬酸的免疫学方法。以活泼酯法将双氯芬酸分别连接到不同载体蛋白制备免疫抗原和检测抗原,免疫家兔制备多克隆抗体,经条件优化建立竞争抑制酶联免疫检测方法。结果显示:检测体系IC50值为9.3ng/mL,最低检出限为2.0ng/mL,与8种相关药物交叉反应率都小于0.1%,用免疫学方法与HPLC检测市场采集的抗风湿保健酒,2例阳性和23例阴性结果全部吻合。该方法灵敏高、特异强,适用于违法添加双氯芬酸食品的初步筛选。
To develop an immunoassay for rapid detection of diclofenac as illegal adulterant in anti-rheumatism health spirit,diclofenac was conjugated to different carrier proteins using active ester method for synthesis of immunogen and testing antigen,respectively.Polyclonal antibodies were obtained from immunized rabbits.Performance of the optimal ELISA was as follows ： the IC 50 value and LOD of the immunoassay were 9.3ng / mL and 2.0ng / mL,respectively.The cross-reactivities against 8associate drugs were less than 0.1%.The ELISA was in good agreement with LC-UV when detected diclofenac in 2positive and 23negative anti-rheumatism health spirit from market.The current assay can be competence for rapid screening of illegal adulterated diclofenac due to its high sensitivity and specificity.
出处
《食品与机械》
CSCD
北大核心
2013年第4期59-62,共4页
Food and Machinery
基金
香港铭源基金科研项目(编号:2010/188)
广东省自然科学基金项目(编号:S2011040001362)
广东省营养膳食与健康重点实验室开放基金项目(编号:2011K004)
关键词
功能食品
风湿病
保健酒
双氯芬酸
人工抗原
免疫检测
functional foods
rheumatism
health spirit
diclofenac
artificial antigen
immunoassay
