保健酒中舒林酸免疫学检测方法的建立

Development of an Immunoassay Method for Detection of Sulindac in Health Liquor

摘要:

舒林酸是常用非甾体抗炎药物，近期常发现不法生产者在保健酒中添加舒林酸增加治疗效果，这种现象对消费者健康构成威胁。为了建立快速检测舒林酸的免疫学方法，本文以活泼酯法将舒林酸分别连接到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18.1 ng/mL，最低检测限为3.5 ng/mL，检测保健酒中舒林酸加标回收率在82.1~94.2%，变异系数小于7.6%。与8种相关药物交叉反应率都小于0.05%。与用ELISA方法与HPLC检测市售保健酒，2例阳性和28例阴性结果全部吻合。因此，本文所建立的舒林酸免疫学检测方法具有灵敏度高和特异性强的特点，能够满足保健酒中快速筛选舒林酸的需要。

Abstract:

lindac is a nonsteroidal anti-inflammatory drug. Recently, it has been found that undeclared sulindac was frequently added into antirheumatic health liquors to promote therapeutic effect, which poses a serious threat to public health. In order to develop an immunoassay for detection of sulindac, providing an analytical method for rapid screening of illegal products, sulindac 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 18.1 ng/L and 3.5 ng/mL, respectively. The recoveries were 76~16% and the coefficients of variation (CV%) were less than 7.6% when detected sulindac in health liquors. All cross-reactivities against 8 associate drugs were less than 0.05%. The ELISA was in good agreement with LC-UV when detecting sulindac in 2 positive and 28 negative health liquors from market. The method was suitable for screening sulindac as illegal additives in health liquors due to its sensitivity and specificity.