肉眼读取-荧光微球竞争免疫层析法半定量快速检测保健食品中的西布曲明  

Fluorescent Microspheres Competitive Immunochromatography Combined with Naked Eye Reading for Rapid and Semi-quantitative Detection of Sibutramine in Food Supplements

作　　者：谭贵良 潘子强[1] 张世伟[2] 胡敏 冯荣虎[2] 杨懋勋 TAN Guiliang;PAN Ziqiang;ZHANG Shiwei;HU Min;FENG Ronghu;YANG Maoxun(Zhongshan Institute,University of Electronic Science and Technology of China,Zhongshan 528402,China;Shenzhen Academy of Metrology and Quality Inspection,Shenzhen 518102,China;Guangdong Institute of Eco-environmental Science&Technology,Guangzhou 510650,China;Guangdong Jiangmen Chinese Medical College,Jiangmen 529030,China)

机构地区：[1]电子科技大学中山学院,广东中山528402 [2]深圳市计量质量检测研究院,广东深圳518102 [3]广东省生态环境技术研究所,广东广州510650 [4]广东江门中医药职业学院,广东江门529030

出　　处：《食品科学》2021年第8期313-318,共6页Food Science

基　　金：中山市社会公益重大项目(2018B1025);广东省自然科学基金项目(2020A1515011308);广东省教学质量与教学改革工程建设项目(SJD202001)。

摘　　要：建立一种新型的西布曲明半定量检测方法。通过在检测试纸条上设置4条包被有抗原的检测线,和被测抗原反应后的剩余标记抗体与包被抗原的反应浓度可梯度降低,实现在单一试纸条构建多级灵敏度的免疫反应,从而建立基于波长470 nm手电筒作为激发光源、肉眼读取结果的西布曲明荧光微球竞争免疫层析半定量检测方法。对吐温-20表面活性剂含量、标记抗体量、包被抗原质量浓度、硝酸纤维膜类型和层析反应时间等参数进行优化,通过试纸条上显色条带的数量对西布曲明进行半定量。结果显示,在优化的实验条件下,该检测方法检出限为0.05μg/mL,检测范围为0~9.00μg/mL。该方法无需使用专门的检测仪器,可在15 min内完成检测,其结果与液相色谱-串联质谱法检测结果一致。该方法操作简便、灵敏度高、结果可靠,可用于保健食品中西布曲明非法添加物的定性和半定量筛查分析。In this study,a novel semi-quantitative method to determine sibutramine in food supplements was developed using fluorescent microspheres competitive immunochromatography coupled with naked eye reading.In the method,four antigen-coated test lines were set on the test strip,allowing decreasing concentration gradients of the labelled antibody left after reaction with the tested antigen to react with the coating antigen,so that immune reaction could be achieved with multiscale sensitivity.The excitation light source was electric torch at 470 nm.The concentrations of Tween-20,labeled antibody and coating antigen,nitrocellulose(NC)membrane type and reaction time were optimized.Semi-quantitative analysis was performed by counting the number of colored test lines.Under the optimized reaction conditions,the sensitivity of the method was 0.05μg/mL and the detection ranged from 0 to 9.00μg/mL.This method took 15 minutes without the need for any special instrument,giving results consistent with those of high performance liquid chromatography-tandem mass spectrometry(LC-MS/MS).The newly developed method was a simple,reliable and rapid method,and could be used in the rapid screening of sibutramine in food supplements.

关 键 词：西布曲明 半定量 荧光微球 免疫层析 保健食品 

分 类 号：TS207.3[轻工技术与工程—食品科学]