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基于适配体技术对止咳平喘健康产品中非法添加的沙丁胺醇进行可视化快速检测
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作者单位
董嘉媚,刘亚雄,樊洪利,罗卓雅 1.广州中医药大学中医药数理工程研究院2.广东省药品检验所国家药品监督管理局药品快速检验技术重点实验室 
基金项目:广东省药品监督管理局创新专项资金项目(ZA20180046,ZA20180049)
中文摘要:基于偶联作用和ssDNA结合蛋白与适配体的结合作用制备检测复合物MB-aptamer~SiO2@SSB@HRP,利用目标物沙丁胺醇与适配体之间的亲和性比ssDNA结合蛋白与适配体亲和性更高的原理,以沙丁胺醇为“钥匙”将适配体与ssDNA结合蛋白构成的“开关”打开,经过磁性分离,最终以辣根过氧化物酶催化四甲基联苯胺产生的显色反应作为信号输出,建立了沙丁胺醇的可视化快速检测方法。在优化实验条件下,沙丁胺醇浓度与显色反应的吸光度在 0.1~10 μmol/L之间呈良好线性关系(r2=0.995 9),检出限为 0.1 μmol/L。对中成药样品进行沙丁胺醇加标回收实验,回收率为860%~106%,相对标准偏差为3.9%~12%。该方法能够满足非法添加实际检测需求。
中文关键词:沙丁胺醇  适配体  可视化快速检测  非法添加  止咳平喘健康产品
 
Rapid Visual Screening of Illegally Added Salbutamol in Health Products Relieving Cough and Asthma by Colorimetric Method Based on Aptamer
Abstract:A rapid visual method based on aptamer sensor was established for screening of illegally added salbutamol(SAL) in health products.The detection element of this sensor was the SAL-aptamer,and the affinity between SAL-aptamer and ssDNA-binding proteins(SSB) acted as a “switch” of the sensor.SAL-aptamer modified micromagnatic beads(MB)(MB-aptamer),and horseradish peroxidase(HRP) and SSB jointly modified silica microsphere(SiO2@SSB@HRP) were prepared,respectively.Then MB-aptamer and SiO2@SSB@HRP were connected by the affinity between SAL-aptamer and SSB.SAL,as a key,triggered the “switch” to result in disconnection between SSB and SAL-aptamer during the inspection process,and the free SiO2@SSB@HRP was separated by magnetic action to carry out a color reaction.Under the optimized conditions,the color reaction showed visible gradient variation according to the change of SAL concentration.The color reaction catalyzed by HRP was quantitatively characterized with an ultraviolet-visible spectrophotometer,there was a linear relationship for absorbance wtih SAL concentration in the range of 0.1-10 μmol/L(r2=0.995 9),with a detection limit of 0.1 μmol/L.The mean recoveries at three spiked levels were all in the range of 86.0%-106%,with relative standard deviations of 3.9%-12%.The method is simple,sensitive and rapid,and it could meet the requirements for the rapid screening of illegally added SAL in real samples.
Key Words:salbutamol  aptamer  rapid visual screening method  illegal addition  health products relieving cough and asthma
引用本文:董嘉媚,刘亚雄,樊洪利,罗卓雅.基于适配体技术对止咳平喘健康产品中非法添加的沙丁胺醇进行可视化快速检测[J].分析测试学报,2020,39(3):383-388.
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