保健酒中萘普生免疫学检测方法的建立

Development of an Immunoassay for Detection of Naproxen in Health Liquor

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。

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 detection 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 detecting Naproxen in health liquors. All cross-reactivities against 8 associate drugs were less than 0.05%. The ELISA method was in good agreement with LC-UV when detecting Naproxen in 1 positive and 19 negative health liquors from market. The method was suitable for screening Naproxen as illegal additives in health liquors due to its sensitivity and specificity.