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).
作者
华修德
尤红杰
杨家川
施海燕
王鸣华
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)
出处
《分析化学》
SCIE
EI
CAS
CSCD
北大核心
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~~
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
Imidaclothiz
Monoclonal antibody
Pesticide residue
Upconversion immunochromatographic assay
Upconversion nanoparticles
