无标记表面等离子共振免疫传感快速测定动物尿液中沙丁胺醇残留  

Label-Free Surface Plasmon Resonance Immunosensor for Rapid Determination of Salbutamol Residue in Animal Urine

作　　者：肖志明[1] 孙铭雪 姚婷 李彦伸[2] 黄骅 王石[1] 索德成[1] 樊霞[1] XIAO Zhiming;SUN Mingxue;YAO Ting;LI Yanshen;HUANG Hua;WANG Shi;SUO Decheng;FAN Xia(China National Feed Quality Inspection and Testing Center(Beijing),Institute of Quality Standard and Testing Technology for Agro-Products,Chinese Academy of Agricultural Sciences,Beijing 100081,China;College of Life Science,Yantai University,Yantai 264005,China;Beijing Veterinary Medicine and Feed Control Center,Beijing 102200,China)

机构地区：[1]中国农业科学院农业质量标准与检测技术研究所,国家饲料质量检验检测中心(北京),北京100081 [2]烟台大学生命科学学院,烟台264005 [3]北京市兽药饲料监测中心,北京102200

出　　处：《动物营养学报》2023年第6期3991-4000,共10页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：“十三五”国家重点研发计划项目(2019YFE0103800)。

摘　　要：本研究旨在建立一种无需标记的直接竞争表面等离子共振(SPR)免疫传感器,以快速测定动物尿液中沙丁胺醇的残留。试验首先利用氨基偶联原理,将沙丁胺醇单克隆抗体固定到CM7传感器芯片表面,然后通过优化单抗固定浓度、pH和固定时间,以及芯片活化和封闭时间,进而得到最佳分析条件。结果表明:沙丁胺醇在0.1~6.0 ng/mL浓度范围内线性关系良好[相关系数(R2)为0.9998],最低检出限(LOD)可达0.01 ng/mL,在添加浓度为0.5、1.0和5.0 ng/mL时,方法的平均回收率为87.82%~97.67%,批内变异系数(相对标准偏差)为1.51%~2.67%,批间变异系数为2.86%~5.53%。将该SPR免疫传感器与超高效液相色谱-串联质谱法(UPLC-MS/MS)确证分析方法进行实际尿液样品测定比较,结果能够保持一致。本研究建立的直接竞争SPR免疫传感方法简便、快速、灵敏,且无需复杂的生物标记,适用于动物尿液中沙丁胺醇残留的日常检测分析。The aim of this study was to establish a direct competitive surface plasmon resonance(SPR)immu⁃nosensor without labeling to rapidly determine albuterol residues in animal urine.Firstly,the amino coupling principle was used to fix salbutamol monoclonal antibody to the surface of the CM7 sensor chip,and then the optimal analysis conditions were obtained by optimizing the fixed concentration,pH and time of the monoclonal antibody,as well as the activation and sealing time of the chip.The results show that the linear relationship of salbutamol was good in the concentration range of 0.1 to 6.0 ng/mL[correlation coefficient(R2)was 0.9998],and the lowest detection limit(LOD)was up to 0.01 ng/mL.The average recovery of salbutamol was 87.82%to 97.67%when the concentrations were 0.5,1.0 and 5.0 ng/mL.The coefficient of variation(relative standard deviation)with intra⁃day was 1.51%to 2.67%,and the coefficient of variation with inter⁃day was 2.86%to 5.53%.The results of the SPR immunosensor were consistent with those of the ultra⁃high performance liquid chromatography⁃tandem mass spectrometry(UPLC⁃MS/MS)confirmations.The direct competition SPR immunosensing method established in this study is simple,rapid and sensitive without com⁃plex biomarkers,and is suitable for the daily detection and analysis of albuterol residues in animal urine.

关 键 词：表面等离子共振 免疫传感器 动物尿液 沙丁胺醇 

分 类 号：S816[农业科学—饲料科学]