基于荧光纳米颗粒的免疫层析技术同步快速检测药食同源药材中的3种真菌毒素  

Simultaneous and Rapid Detection of Three Mycotoxins in Food and Medicine Homologous Materials by Fluorescent Nanoparticle-Based Immunochromatography

作　　者：霍炳帆 马骉 厉佳丽 梅青 周宇欣 张明洲 HUO Bingfan;MA Biao;LI Jiali;MEI Qing;ZHOU Yuxin;ZHANG Mingzhou(College of Life Sciences,China Jiliang University,Hangzhou 310018,China;Hangzhou Kuige Technology Co.Ltd.,Hangzhou 310002,China)

机构地区：[1]中国计量大学生命科学学院,浙江杭州310018 [2]杭州快格科技有限公司,浙江杭州310002

出　　处：《现代食品科技》2024年第7期227-235,共9页Modern Food Science and Technology

基　　金：浙江省重点研发计划项目(2021C02060,2020C02024);浙江省基础公益研究计划农业农村项目(LGN22C200015)。

摘　　要：该研究建立了一种基于铕纳米粒子(Europium Nanoparticles,EuNPs)的超灵敏侧向流动免疫层析(Lateral Flow Immunochromatography,LFIC)方法,用于药食同源药材中黄曲霉毒素B1(Aflatoxin B1,AFB1)、玉米赤霉烯酮(Zearalenone,ZEN)和赭曲霉毒素A(Ochratoxin A,OTA)的同步快速检测。通过优化人工抗原包被质量浓度、人工抗原划膜位置、羊抗鼠IgG包被质量浓度、缓冲液pH值和免疫反应时间等关键因素,在简单的样品前处理步骤后,能够实现单个待测样品中3种目标真菌毒素的超灵敏联合检测。结果显示,该方法特异性良好,灵敏度高,AFB1、ZEN和OTA标准品的肉眼检测限分别为0.5、4.0、2.0 ng/mL,线性范围(IC10~IC90)分别为0.03~4.61、0.12~8.47、0.09~8.26 ng/mL。该方法用于人参、黄芪和当归中真菌毒素的测定,样品平均添加回收率为90.02%~109.39%,相对标准偏差(RSD)小于8.50%,实际样品检测结果与高效液相色谱法符合率达100%。因此,该研究构建的EuNPs-LFIC方法适用于药食同源药材中的AFB1、ZEN与OTA等真菌毒素的现场快速监测。An ultra-sensitive lateral flow immunochromatography(LFIC)method based on europium nanoparticles(EuNPs)was established for the simultaneous and rapid detection of aflatoxin B1(AFB1),zearalenone(ZEN),and ochratoxin A(OTA)in medicine and food homologous materials.By optimizing the key parameters such as the mass concentration of the coated artificial antigen, immobilized line position of the coated artificial antigen, mass concentration of the coated goat anti- mouse IgG, buffer pH, and immunoreaction time, the developed method could achieve the ultrasensitive and simultaneous identification of three target mycotoxins in one sample after a simple sample pretreatment. This method demonstrated a high specificity and sensitivity. The visual limits of detection of the AFB1, ZEN, and OTA standards were 0.5, 4.0, and 2.0 ng/mL, respectively. Their linear ranges (IC10~IC90) were 0.03~4.61, 0.12~8.47, and 0.09~8.26 ng/mL, respectively. In the detection of mycotoxins in ginseng, astragalus, and angelica samples, the mean spiked recovery rates of the three types of analytes ranged from 90.02% to 109.39%, with a relative standard deviation of less than 8.50%. The coincidence rate between the actual sample and HPLC method was 100%. Therefore, the developed EuNP-LFIC method is suitable for the on-site rapid monitoring of mycotoxins such as AFB1, ZEN, and OTA in medicine and food homologous materials.

关 键 词：铕纳米粒子 侧向流动免疫层析 真菌毒素 同时检测 

分 类 号：O657.3[理学—分析化学] R284.1[理学—化学]