基于适配体/金纳米粒子的“开-关”型荧光探针快速测定水胺硫磷

引用本文: 邹小波, 蒋彩萍, 李志华, 孙悦, 石吉勇, 李艳肖, 黄晓玮, 张迪, 胡雪桃, 翟晓东, 魏晓鸥. 基于适配体/金纳米粒子的“开-关”型荧光探针快速测定水胺硫磷[J]. 分析化学, 2020, 48(4): 507-515. doi: 10.19756/j.issn.0253-3820.191335 shu

Citation: ZOU Xiao-Bo, JIANG Cai-Ping, LI Zhi-Hua, SUN Yue, SHI Ji-Yong, LI Yan-Xiao, HUANG Xiao-Wei, ZHANG Di, HU Xue-Tao, ZHAI Xiao-Dong, WEI Xiao-Ou. A “Turn on-off” Fluorescent Probe Based on Aptamer/Gold Nanoparticles for Rapid Detection of Isocarbophos[J]. Chinese Journal of Analytical Chemistry, 2020, 48(4): 507-515. doi: 10.19756/j.issn.0253-3820.191335 shu

基于适配体/金纳米粒子的“开-关”型荧光探针快速测定水胺硫磷

邹小波 ^1, ,
蒋彩萍 ^1, ,
李志华 ^1, ,
孙悦 ^1, ,
石吉勇 ^1, ,
李艳肖 ^2, ,
黄晓玮 ^1, ,
张迪 ^1, ,
胡雪桃 ^1, ,
翟晓东 ^1, ,
魏晓鸥 ^1,

1.
江苏大学食品与生物工程学院, 镇江 212013;
2.
江苏大学分析测试中心, 镇江 212013
基金项目:
本文系国家重点研发计划项目（Nos.2017YFC1600806，2016YFD0401104）、国家自然科学基金项目（Nos.31801631，31671844，31601543）、江苏省自然科学基金（Nos.BK20160506，BE2016306，BK20180865）和江苏省国际科技合作项目（No.BZ2016013）资助

摘要: 基于适配体和金纳米粒子（Gold nanoparticles，AuNPs）合成"开-关"型荧光探针，用于快速、灵敏、高选择性测定水胺硫磷。5'端连接荧光素（6-Carboxyfluorescein，FAM）的发卡型水胺硫磷适配体互补链（Complementary strand，cDNA）通过修饰于3'端的巯基连接到AuNPs表面，FAM（荧光信号分子）与AuNPs（荧光猝灭剂）间发生荧光共振能量转移效应（Fluorescence resonance energy transfer，FRET），导致荧光猝灭。加入水胺硫磷适配体（Isocarbophos-binding aptamer，ICP-Aptamer）与cDNA杂交，cDNA发卡构型被打开，使荧光信号恢复。当体系中存在水胺硫磷时，ICP-Aptamer与其特异性结合，导致与cDNA解离，cDNA发卡型结构重新恢复，使荧光信号猝灭。采用透射电镜、紫外-可见光谱、Zeta电位表征纳米粒子特性，优化了实验参数，包括pH值、ICP-Aptamer浓度、温育时间和温育温度。在优化的实验条件下，在0.02~10 μmol/L线性范围内，水胺硫磷浓度与荧光抑制率呈良好的线性关系，检出限为17.8 nmol/L（3σ）。将此荧光探针应于大米、菠菜样品中水胺硫磷的测定，回收率均为92.9%~107.0%，相对标准偏差小于4.2%，表明此荧光探针在农药的检测和食品安全的监测中具有潜在的应用价值。

关键词:

English

A “Turn on-off” Fluorescent Probe Based on Aptamer/Gold Nanoparticles for Rapid Detection of Isocarbophos

1.
School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, China;
2.
Analytical Instrumentation Center, Jiangsu University, Zhenjiang 212013, China
Received Date: 14 June 2019
Revised Date: 21 February 2020
Fund Project:
This work was supported by the National Key Research and Development Program of China (Nos. 2017YFC1600806, 2016YFD0401104), the National Natural Science Foundation of China (Nos. 31801631, 31671844, 31601543), the Natural Science Foundation of Jiangsu Province (Nos. BK20160506, BE2016306, BK20180865) and the International Science and Technology Cooperation Project of Jiangsu Province (No. BZ2016013).

Abstract: A "turn on-off" fluorescent probe based on aptamer/gold nanoparticles(AuNPs) was developed for rapid, highly sensitive and selective detection of isocarbophos (ICP). Hairpin complementary strand (cDNA) with a fluorescein label (FAM) at 5' end was attached to the surface of AuNPs through thiol group. Subsequently, ICP-binding aptamer (ICP-Aptamer) was added and hybridized with cDNA, leading to open of hairpin assembly and "turn on" of fluorescent signal quenched by AuNPs. ICP-Aptamer specifically bound to the target ICP with restoration of configuration and "turn off" of the fluorescent signal. The properties of nanoparticles were characterized by transmission electron microscopy, ultraviolet-visible absorption spectroscopy and zeta potential. Experimental parameters influencing fluorescence inhibition rate, including pH value, concentration of ICP-Aptamer, incubation time and incubation temperature, were investigated and optimized. The corresponding fluorescence inhibition rate had a good linear relationship with concentration of ICP in the range of 0.02-10 μmol/L, and the limit of detection was 17.8 nmol/L. Furthermore,the fluorescent probe was applied to determination of ICP in rice and spinach samples. The recoveries were from 92.9% to 107.0% and the relative standard deviations were lower than 4.2%. It could be concluded that the fluorescence probe had good application potential for detection of pesticides and supervision of food safety.

Key words:

1. [1]
  Pundir C S, Malik A, Preety. Biosens. Bioelectron., 2019, 140: 111348
2. [2]
  Qiu L H, Lv P, Zhao C L, Feng X Y, Fang G Z, Liu J F, Wang S. Sens. Actuators B, 2019, 286: 386-393
3. [3]
  Jia M, Wang Y, Teng M M, Wang D Z, Yan J, Miao J Y, Zhou Z Q, Zhu W T. Ecotox. Environ. Safe., 2018, 163: 1-6
4. [4]
  GB 24753-2009, Isocarbophos Technical. National Standards of the People's Republic of China 水胺硫磷原药. 中华人民共和国国家标准. GB 24753-2009
5. [5]
  Timofeeva I, Shishov A, Kanashina D, Dzema D, Bulatov A. Talanta, 2017, 167: 761-767
6. [6]
  Taha S M, Gadalla S A. Talanta, 2017, 174: 767-779
7. [7]
  Zhang C, Cui H Y, Han Y F, Yu F F, Shi X M. Food Chem., 2018, 240: 893-897
8. [8]
  Senyuva H Z, Gilbert J. J. Chromatogr. B, 2010, 878 (2): 115-132
9. [9]
  Pan J F, Li Q, Zhou D H, Chen J H. Talanta, 2018, 189: 370-376
10. [10]
  Sun Y, Li Z H, Huang X W, Zhang D, Zou X B, Shi J Y, Zhai X D, Jiang C P, Wei X O, Liu T T. Biosens. Bioelectron., 2019, 145, 111672
11. [11]
  Fu J Y, An X S, Yao Y, Guo Y M, Sun X. Sens. Actuators B, 2019, 287: 503-509
12. [12]
  Jia L P, Zhou Y X, Wu K P, Feng Q L, Wang C M, He P. Bioelectrochemistry, 2020, 131: 107392
13. [13]
  Pérez-Fernández B, Mercader J V, Abad-Fuentes A, Checa-Orrego B I, Costa-Garcia A, de la Escosura-Muñiz A, Costa-García A. Talanta, 2019, 209: 120465
14. [14]
  Hu X T, Shi J Y, Shi Y Q, Zou X B, Tahir H E, Holmes M, Zhang W, Huang X W, Li Z H, Xu Y W. Meat Sci., 2019, 147: 127-134
15. [15]
  ZOU Xiao-Bo, SHI Yong-Qiang, ZHENG Yue, SHI Ji-Yong, HU Xue-Tao, JIANG Cai-Ping, HUANG Xiao-Wei, XU Yi-Wei. Chiness J. Anal. Chem., 2018, 46(6): 960-968 邹小波, 史永强, 郑悦, 石吉勇, 胡雪桃, 蒋彩萍, 黄晓玮, 徐艺伟. 分析化学, 2018, 46(6): 960-968
16. [16]
  Zhang W, Xu Y W, Zou X B. Food Chem., 2018, 261: 1-7
17. [17]
  Li Z H, Zhou X C, Wang K, Zou X B, Shi J Y, Huang X Y, Holmes M. Innov. Food Sci. Emerg., 2015, 31: 196-203
18. [18]
  Li Z H, Zhou X C, Shi J Y, Zou X B, Huang X Y, Tahir H E. Food Chem., 2019, 276: 291-297
19. [19]
  Xu X C, Niu X H, Wu S W, Zou X B, Pan J M. Sens. Actuators B, 2018, 268: 430-437
20. [20]
  ZHANG Jing-Jing, KANG Tian-Fang, LU Li-Ping, CHENG Shui-Yuan. Anal. Chem., 2016, 44 (5): 760-766 张晶晶, 康天放, 鲁理平, 程水源. 分析化学, 2016, 44 (5): 760-766
21. [21]
  ZOU Xue-Mei, ZHOU Jia-Wei, SONG Shang-Hong, CHEN Guan-Hua. Anal. Chem., 2019, 47 (4): 488-499 邹雪梅, 周佳伟, 宋尚红, 陈冠华. 分析化学, 2019, 47 (4): 488-499
22. [22]
  Taghdisi S M, Danesh N M, Lavaee P, Ramezani M, Abnous K. Sens. Actuators B, 2016, 234: 462-469
23. [23]
  Emrani A S, Danesh N M, Ramezani M, Taghdisi S M, Abnous K. Biosens. Bioelectron., 2016, 79: 288-293
24. [24]
  Zhang C Z, Wang L, Tu Z, Sun X, He Q H, Lei Z J, Xu C X, Liu Y, Zhang X, Yang J Y, Liu X J, Xu Y. Biosens. Bioelectron., 2014, 55: 216-219
25. [25]
  Turkevich J, Stevenson P C, Hillier J. Discuss. Faraday Soc., 1951, 11: 55-75
26. [26]
  Tang T T, Deng J J, Zhang M, Shi G Y, Zhou T S. Talanta, 2016, 146, 55-61
27. [27]
  Yao G B, Pei H, Li J, Zhao Y, Zhu D, Zhang Y N, Lin Y F, Huang Q, Fan C H. NPG Asia Mater., 2015, 7 (1): e159
28. [28]
  Haiss W, Thanh N T K, Aveyard J, Fernig D G. Anal. Chem., 2007, 79 (11): 4215-4221
29. [29]
  Asati A, Santra S, Kaittanis C, Perez J M. ACS Nano, 2010, 4 (9): 5321-5331
30. [30]
  Liu B Y, Zeng F, Wu S Z, Wang J S, Tang F C. Microchim. Acta, 2013, 180 (9): 845-853
31. [31]
  Hu X T, Shi J Y, Shi Y Q, Zou X B, Arslan M, Zhang W, Huang X W, Li Z H, Xu Y Y. Food Chem., 2019, 272: 58-65
32. [32]
  Wang L, Li H, Yang Y, Zhang D, Wu M, Pan B, Xing B S. Water Res., 2017, 122: 337-344
33. [33]
  WANG Li, YE Hua, SANG Hong-Qing, WANG Dan-Dan. Chinese J.Anal. Chem., 2016, 44 (5): 799-803 王丽, 叶华, 桑宏庆, 王丹丹. 分析化学, 2016, 44 (5): 799-803
34. [34]
  Yan X N, Deng J, Xu J S, Li H, Wang L L, Chen D, Xie J. Sens. Actuators B, 2012, 171-172: 1087-1094
35. [35]
  Shi Q W, Teng Y J, Zhang Y C, Liu W H. Chinese Chem. Lett., 2018, 29 (9): 1379-1382
36. [36]
  Pang S, Labuza T P, He L L. Analyst, 2014, 139: 1895-1901
37. [37]
  Bai W H, Zhu C, Liu J C, Yan M M, Yang S M, Chen A L. Environ. Toxicol.Chem., 2015, 34 (10): 2244-2249

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基于适配体/金纳米粒子的“开-关”型荧光探针快速测定水胺硫磷

English

A “Turn on-off” Fluorescent Probe Based on Aptamer/Gold Nanoparticles for Rapid Detection of Isocarbophos

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

基于适配体/金纳米粒子的“开-关”型荧光探针快速测定水胺硫磷

English

A “Turn on-off” Fluorescent Probe Based on Aptamer/Gold Nanoparticles for Rapid Detection of Isocarbophos

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

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CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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