基于上转换荧光标记的氯噻啉免疫层析方法研究  

Immunochromatographic Assay for Detection of Imidaclothiz Based on Upconversion Fluorescence Labeling

作　　者：华修德[1] 尤红杰 杨家川 施海燕[1] 王鸣华[1] HUA Xiu-De;YOU Hong-Jie;YANG Jia-Chuan;SHI Hai-Yan;WANG Ming-Hua(College of Plant Protection,Nanjing Agricultural University,Nanjing 210095,China)

机构地区：[1]南京农业大学植物保护学院,南京210095

出　　处：《分析化学》2018年第3期413-421,共9页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金项目(No.31471794)资助~~

摘　　要：利用氯噻啉单克隆抗体标记的Na YF4:Yb,Er上转换荧光纳米材料(Upconversion nanoparticles,UCNPs)建立了一种简单、快速、灵敏的氯噻啉上转换荧光免疫层析(Upconversion immunochromatographic assay,UICA)方法。将氨基功能化的UCNPs与氯噻啉单克隆抗体偶联制备UICA试纸条,使用外接980 nm激光光源的荧光分光光度计实现对氯噻啉的定量检测。在优化的UICA工作条件下,通过交叉反应(Crossreactivity,CR)、添加回收和高效液相色谱(High performance liquid chromatography,HPLC)实验评价了UICA的敏感性、特异性、精确性和准确性。在最优条件(p H 8.0、0.3 mol/L Na Cl、2.5%甲醇、0.2%PEG2000)下,UICA可在25 min内完成对氯噻啉的检测,其抑制中浓度(Half-maximal inhibition concentration,IC_(50))为97.37 ng/m L,检出限(IC_(10))为26.30 ng/m L,线性范围(IC_(10)~IC_(90))为26.30~363.08 ng/m L。除吡虫啉外,UICA对其它氯噻啉结构类似物无交叉反应。在田水、土壤、梨、桃、小麦、黄瓜、番茄、大米等基质中的平均添加回收率为71.8%~97.2%,相对标准偏差为0.7%~10.7%。UICA对田水和梨实际样品的检测结果与HPLC的检测结果相符。A simple,rapid and sensitive upconversion immunochromatographic assay(UICA)was developed to detect imidaclothiz using NaYF4:Yb,Er upconversion nanoparticles(UCNPs)labeled with anti-imidaclothiz monoclonal antibody.The amino-modified UCNPs were conjugated with anti-imidaclothiz monoclonal antibody to prepare the UICA strip,which could realize the quantitative detection of imidaclothiz using a fluorescence photometer with an external 980 nm laser source.The working conditions of the UICA were systematically optimized,and the sensitivity,specificity,precision and accuracy were assessed by the studies of cross-reactivity(CR),spiked recovery and validation with HPLC.Under the optimal conditions(pH 8.0,0.3 mol/L NaCl,2.5%methanol and 0.2%PEG2000),the UICA could be completed in 25 min for the detection of imidaclothiz.The half-maximal inhibition concentration(IC50),limit of detection(IC10)and linear range(IC10-IC90)were 97.37 ng/mL,26.30 ng/mL and 26.30-363.08 ng/mL,respectively.The UICA had no CR with the analogues of imidaclothiz except for imidacloprid.The average spiked recoveries were 71.8%-97.2%with the relative standard deviations of 0.7%-10.7%in the matrices of paddy water,soil,pear,peach,wheat,cucumber,tomato and rice.The detection results of UICA for the authentic paddy water and pear samples were consistent with that of high performance liquid chromatography(HPLC).

关 键 词：氯噻啉 单克隆抗体 农药残留 上转换荧光免疫层析 上转换荧光纳米材料 

分 类 号：O658.1[理学—分析化学] TQ450.263[理学—化学]