保健酒中萘普生免疫学检测方法的建立.pdfVIP

现代食品科技 Modern Food Science and Technology 2013, Vol.29, No.8 保健酒中萘普生免疫学检测方法的建立 郭杰标，郝卿辰，李杏娉，肖仔君，何颖娟，刘艳珊 (韶关学院，广东韶关 512005) 摘要：萘普生是疗效显著但有一定副作用的非甾体抗炎药物，近期频繁发现不法生产者在抗风湿保健酒中添加萘普生增加治疗 效果，这种现象对消费者健康构成威胁。为了究建立快速检测萘普生的免疫学方法，本文以活泼酯法将萘普生分别连接到BSA 制备 免疫抗原，连接到OVA 上制备检测抗原。用免疫抗原免疫家兔制备多克隆抗体，配合检测抗原经条件优化建立竞争抑制酶联免疫吸 附检测方法。实验结果表明，检测体系IC50 值为23.0 ng/mL，最低检测限为3.1 ng/mL，检测保健酒中萘普生加标回收率在87.3~102.1%， 变异系数小于8.9%，与8 种相关药物交叉反应率都小于0.05% 。且与用ELISA 方法与HPLC 检测市售保健酒，1 例阳性和 19 例阴性 结果全部吻合。因此，本文所建立的免疫学检测萘普生的实验方法，具有高灵敏度和特异性，能够满足保健酒萘普生快速筛选的需要。 关键词：功能食品；萘普生；违法添加；人工抗原；免疫检测 文章篇号：1673-9078(2013)8-2011-2014 Development of an Immunoassay for Detection of Naproxen in Health Liquor GUO Jie-biao, HAO Qing-chen, LI Xing-ping, XIAO Zi-jun, HE Ying-juan, LIU Yan-shan (Shaoguan College, Shaoguan 512005, China) Abstract: Naproxen,a nonsteroidal anti-inflammatory drug, was frequently reported being adulterated onto antirheumatic health liquors to promote therapeutic effect. This phenomenon poses a serious threat to public health. In order to develop an immunoassay for detect ion of naproxen, providing an analytical method for rapid screening of illegal products, naproxen was conjugated to BSA and OVA to synthesize artificial antigens using active ester method for preparation of immunogen and testing antigen, respectively. Immunogen was employed to immunize rabbits for raising polyclonal antibodies. An enzyme-linked immunosorbent assay (ELISA) was established based on the antibodies and testing antigen. The IC50 value and LOD of the optimal ELISA were 23.0 ng/mL and 3. 1 ng/mL, respectively. The recoveries were 87.3 ~102.1% and the coefficients of variation (CV%) were less than 8.9% when detect ing Na