【作者】 祝春节； 俞琼琼； 赵广英； 窦文超；

【Author】 Zhu Chunjie;Yu Qiongqiong;Zhao Guangying;Dou Wenchao;Zhejiang gongshang university school of food science and biotechnology;

【机构】 浙江工商大学食品与生物工程学院；

【摘要】 为研制新型标记物并用于改善盐酸克伦特罗免疫层析试纸快速检测技术（immunochromatographic assay,ICA）,本研究采用Stber法,首次以活性紫（C.I.Reactive violet 5）为模型染料制备紫色SiO₂纳米颗粒（pmpie SiNPs）,并优化硅烷偶联剂（3APTMS）和活性紫染料的剂量对purple SiNPs分散性和染料偶联率的影响。优化条件下,球形规则、粒径均一、分散性好,且制备过程环保、经济。继而首次将其作为标记物,构建以盐酸克伦特罗（Clebuterol,CLE）为研究模型的小分子药物免疫层析竞争法。优化条件下,对PBS中CLE的检测灵敏度为3 ng/mL,对猪尿液和猪肉加标样品中CLE的检测灵敏度为6 ng/mL和5 ng/mL,可见灵敏度很高。检测他种4种相似结构的小分子标准品无交叉反应,表明其具有良好的特异性。该免疫层析试纸条室温干燥存放2个月后可正常使用,表明稳定性好。由此可见,该方法有良好的灵敏度,特异性和稳定性。因此,purpie SiNPs能作为免疫层析标记探针,用于小分子药物CLE的快速检测,为后续多种目标物的检测提供了良好的研究基础。更多还原

【Abstract】 To prepare a new type of marker and to improve the immunochromatographic assay(ICA) to detect Clebuterol(CLE),in the study,the silica nanoparticles(SiNPs) were synthesized by Stober method,then the C.I.Reactive violet 5 was selected to be the model dye in this work to synthesize purple silica nanoparticles(purple SiNPs),and the effect of the amount of silane coupling agent(3 APTMS) and C.I. Reactive violet 5 dye on preparation of purple SiNPs was studied.Under the optimal conditions,purple SiNPs had regular morphology,uniform size and good dispersibility,and the preparation process is environmentally friendly and economical.Then the purple SiNPs were first applicated as labels for detection of the model of Clebuterol(CLE) of small molecule drug competive ICA.Under the optimal conditions,the detection sensitivity of this purple SiNPs based ICA for CLE was 3 ng/mL in sterile phosphate buffer saline(PBS),and that in swine urine and pork samples were 6 ng/mL and 5 ng/mL,which showed that the sensitivity was very high.The purple SiNPs based ICA has no cross activity with other four nonspecific structural analogs,demonstrating it has good specificity.The strips can be used normally after being stored at room temperature for 2 months,which showed good stability.Therefore,the type immunochromatographic assay based on competive principles proved that purple SiNPs can be used as an alternative visible label for the ICA to detect small molecule drug of CLE,providing a good research foundation for posterior detection of other targets.更多还原