基于智能手机利用蛋白芯片法检测猪肉中克伦特罗和莱克多巴胺的残留量  

Detection of Clenbuterol and Ractopamine Residues in Pork Using Protein Chip Method Based on Smart Phones

作　　者：李周敏[1] 常博文 陈炳丰 李心爱[1] 谷苗苗[1] LI Zhoumin;CHANG Bowen;CHEN Bingfeng;LI Xin’ai;GU Miaomiao(Nanjing University Jinling College,Nanjing 210089,China)

机构地区：[1]南京大学金陵学院,江苏南京210089

出　　处：《食品科技》2021年第10期271-276,共6页Food Science and Technology

基　　金：江苏省高校“青蓝工程”项目(0010392001);南京大学金陵学院教学改革项目(0010522020,0010522021);江苏省大学生创新创业训练项目(202013646058X)。

摘　　要：以智能手机为主要检测设备,Photoshop和Genpix为辅助软件,建立了猪肉中同时检测克伦特罗和莱克多巴胺残留量的蛋白芯片法。采用间接竞争免疫法,固定在孔板底部的人工抗原和溶液中游离的标准品与其相应抗体发生竞争反应。随后加入显色液,生成黑色沉淀,再利用智能手机获取图像,软件处理后得到灰度信号值。根据标准曲线计算样品中克伦特罗和莱克多巴胺的含量。对反应温度、时间、加样体积、转速等主要影响因素进行优化,得出最佳反应条件:37℃,反应30 min,加样体积为50μL/孔,转速为600 r/min。克伦特罗的检测限为0.17 ng/g,莱克多巴胺的检测限为0.39 ng/g。猪肉样品的加标回收率在84.60%~109.67%。基于智能手机利用蛋白芯片法用于实际样品中瘦肉精残留量的检测,该方法只需一步免疫反应,即可实现高通量、多指标同时快速定量检测,且无需专业的检测设备,利于监管部门进行现场执法。In this paper,a protein chip method for simultaneous determination of clenbuterol and ractopamine residues in pork was established with smart phone as the main detection equipment and Photoshop and Genpix as auxiliary software.Using indirect competitive immunity,the artificial antigen fixed at the bottom of the plate and the standard product floating in solution competed with the corresponding antibody.Then added the color developing solution,after the black precipitation was generated,the image was acquired by the smart phone,and the gray signal value was obtained after software processing.Clenbuterol and ractopamine levels were calculated according to the standard curve.Then the main influencing factors such as reaction temperature,reaction time,sample volume,rotation speed and so on were optimized.The optimal reaction conditions were obtained as follows:37℃,reaction time for 30 min,sample volume 50μL/well,rotation speed 600 r/min.The limit of detection was 0.17 ng/g for clenbuterol and 0.39 ng/g for ractopamine.The recoveries of pork samples were 84.60%~109.67%.The protein chip method based on smart phones can be used for the detection of clenbuterol and ractopamine residues in actual samples.This method requires only one step for immune response,high-throughput,can rapid quantitatively detect multiple indicators at the same time,and does not require professional detecting equipment,which is conducive to on-site law enforcement by regulatory agencies.

关 键 词：智能手机 蛋白芯片 克伦特罗 莱克多巴胺 

分 类 号：TS207.53[轻工技术与工程—食品科学]