Citation: WEI Chun-Hao, YANG Guang-Xin, CHI Hai, TAO Le-Ren, KONG Cong, CAI You-Qiong. Inner Filter Effect-based Fluorescence Immunoassay with GoldNanoclusters for Malachite Green Detection[J]. Chinese Journal of Analytical Chemistry, ;2022, 50(1): 73-81. doi: 10.19756/j.issn.0253-3820.201750 shu

Inner Filter Effect-based Fluorescence Immunoassay with GoldNanoclusters for Malachite Green Detection

WEI Chun-Hao ^1,2 ,
YANG Guang-Xin ^1,, ,
CHI Hai ^1,2,, ,
TAO Le-Ren ² ,
KONG Cong ¹ ,
CAI You-Qiong ¹

1.
East China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Shanghai 200090, China;
2.
Schoolof Medical Instrument and Food Engineering, University of Shanghai for Science and Technology, Shanghai 200093, China

Corresponding author: YANG Guang-Xin, CHI Hai,
Received Date: 11 December 2020
Revised Date: 9 June 2021

Fund Project: Supported by Central Public-interest Scientific Institution Based Research Fund (No.2016T09)

Malachite green (MG), an effective insecticide and preservative, is commonly used in the aquaculture industry. However, MG has acute toxicity to terrestrial and aquatic animals, which can lead to mutagenicity, carcinogenicity, and teratogenicity on animals and human bodies. Therefore, it is necessary to explore a reliable and sensitive method for the rapid detection of MG. Herein, a novel and sensitive fluorescence-linked immunosorbent assay (FLISA) was constructed based on glod nanoclusters (BSA-AuNCs) for MG detection. The assay signal was originated from inner filter effect (IFE) between BSA-AuNCs and p-nitrophenol (PNP). Firstly, a double emission peak fluorescent gold nanocluster with good light stability and ultra-small size was quickly synthesized in 7 minutes, which was used as fluorescence probe in this immunoassay. Secondary, alkaline phosphatase labeled secondary antibody (IgG-ALP) was used to replace IgG-HRP in the immune reaction, which could generate PNP with the substrate of p-nitrophenyl phosphate (PNPP). As the maximum excitation wavelength of BSA-AuNCs (390 nm) overlapped with the absorption peak of PNP (400 nm), the fluorescence of BSA-AuNCs could be strongly quenched by PNP. Thus, with the absence of MG in the indirect immunoassay, much more IgG-ALP would be adsorbed in the 96-well plates, more PNP would be produced, then with the fluorescence quenched more strongly. The fluorescence signal could be used for MG detection indirectly. In this immunoassay, several factors including coating antigen, antibody concentration, secondary antibody concentration, substrate concentration, reaction temperature and reaction time were optimized. Under the optimized conditions, the linear range of this method was 0.1-100 ng/mL, linear equation was y=181.53lgC_MG+241.3 (R²=0.995), and the limit of detection (LOD) was 0.089 ng/mL. The sensitivity of FLISA was 30 times that of ic-ELISA. Furthermore, the proposed method was successfully applied to determination of MG in spiked aquaculture water and tap water. The recoveries of MG were in the range of 84.0%-110.2%, and the coefficient of variation was 4.6%-11.2%. The new immunoassay could be used as a promising tool for rapid detection of MG in real environmental samples.
- Malachite green,
- Fluorescence,
- Inner filter effect,
- Gold nanoclusters,
- Enzyme-linked immunosorbent assay
1. [1]
  WU L, LIN Z Z, ZHONG H P, PENG A H, CHEN X M, HUANG Z Y. Food Chem., 2017, 229:847-853.
2. [2]
  SAHRAEI R, FARMANY A, MORTAZAVI S S, NOORIZADEH H. Environ. Monit. Assess., 2013, 185:5817-5822.
3. [3]
  ZHANG Y Y, HUANG Y Q, ZHAI F L, DU R, LIU Y D, LAI K Q. Food Chem., 2012, 135(2):845-850.
4. [4]
  ZHANG K, SONG G, YANG L X. Anal. Methods, 2012, 4(12):4257-4263.
5. [5]
  LIN Z Z, ZHANG H Y, PENG A H, LIN Y D, LI L, HUANG Y. Food Chem., 2016, 200:32-37.
6. [6]
  JIA F, YANG X D, LI Z Y. RSC Adv., 2016, 6:92723-92728.
7. [7]
  XIE J, PENG T, CHEN D D, ZHANG Q J, WANG G M, WANG X, QI G, JIANG F, CHEN D, DENG J. J. Chromatogr. B, 2013, 913:123-128.
8. [8]
9. [9]
10. [10]
11. [11]
  GUO Y R, ZOU R B, SI F F, LIANG W L, ZHANG T Y, CHANG Y Y, QIAO X S, ZHAO J H. Food Chem., 2020, 335:127609-127617.
12. [12]
  ZHAO M T, LI X L, ZHANG Y L, WANG Y W, WANG B, ZHENG L L, ZHANG D W, ZHUANG S L. Food Chem., 2020, 339:127857-127864.
13. [13]
  ZHANG C, JIANG Z J, JIN M J, DU P F, CHEN G, CUI X Y, ZHANG Y D, QIN G X, YAN F Y, EL-ATY A M A, HACIMVFTVOǦLU A, WANG J. Food Chem., 2020, 326:126813-126820.
14. [14]
  CHEN S, YU Y L, WANG J H. Anal. Chim. Acta, 2018, 999:13-26.
15. [15]
  DONG B L, LI H F, MARI G M, YU X Z, YU W B, WEN K, KE Y B, SHEN J Z, WANG Z H. Food Chem., 2019, 294:347-354.
16. [16]
  LUO L, SONG Y, ZHU C Z, FU S F, SHI Q R, SUN Y M, JIA B Z, DU D, XU Z L, LIN Y H. Sens. Actuators, B, 2018, 255(3):2742-2749.
17. [17]
  CUI M L, YUAN Z, SONG Q J. TrAC-Trends Anal. Chem., 2014, 57:73-82.
18. [18]
19. [19]
  XIE J P, ZHENG Y G, YING J Y. J. Am. Chem. Soc., 2009, 131(3):888-889.
20. [20]
  LI H F, WEN K, DONG B L, ZHANG J, BAI Y C, LIU M G, LI P P, MUJTABA G M, YU X Z, YU W B,KE Y B, SHEN J Z, WANG Z H. Sens. Actuators, B, 2019, 297:126787-126796.
21. [21]
22. [22]
  JU Y J, LI N, LIU S G, HAN L, XIAO N, LUO H Q, LI N B. Sens. Actuators, B, 2018, 275:244-250.
23. [23]
  LIU H Y, ZHANG X, WU X M, JIANG L P, BURDA C, ZHU J J. Chem. Commun., 2011, 47(14):4237-4239.
24. [24]
  KONG Y F, CHEN J, GAO F, BRYDSON R, JOHNSON B, HEATH G, ZHANG Y, WU L, ZHOU D J. Nanoscale, 2013, 5(3):1009-1017.
25. [25]
  HSUA N Y, LIN Y W. New J. Chem., 2016, 40(2):1155-1161.
26. [26]
27. [27]
1. [1]
  Feng Lu , Tao Wang , Qi Wang . Preparation and Characterization of Water-Soluble Silver Nanoclusters: A New Design and Teaching Practice in Materials Chemistry Experiment. University Chemistry, 2025, 40(4): 375-381. doi: 10.12461/PKU.DXHX202406005
2. [2]
  Peng XU , Shasha WANG , Nannan CHEN , Ao WANG , Dongmei YU . Preparation of three-layer magnetic composite Fe₃O₄@polyacrylic acid@ZiF-8 for efficient removal of malachite green in water. Chinese Journal of Inorganic Chemistry, 2024, 40(3): 544-554. doi: 10.11862/CJIC.20230239
3. [3]
  YanYuan Jia , Rong Rong , Jie Liu , Jing Guo , GuoYu Jiang , Shuo Guo . Unity is Strength, and Independence Shines: A Science Popularization Experiment on AIE and ACQ Effects. University Chemistry, 2024, 39(9): 349-358. doi: 10.12461/PKU.DXHX202402035
4. [4]
  Chen LU , Qinlong HONG , Haixia ZHANG , Jian ZHANG . Syntheses, structures, and properties of copper-iodine cluster-based boron imidazolate framework materials. Chinese Journal of Inorganic Chemistry, 2025, 41(1): 149-154. doi: 10.11862/CJIC.20240407
5. [5]
  Xiao SANG , Qi LIU , Jianping LANG . Synthesis, structure, and fluorescence properties of Zn(Ⅱ) coordination polymers containing tetra-alkenylpyridine ligands. Chinese Journal of Inorganic Chemistry, 2024, 40(11): 2124-2132. doi: 10.11862/CJIC.20240158
6. [6]
  Qin Li , Kexin Yang , Qinglin Yang , Xiangjin Zhu , Xiaole Han , Tao Huang . Illuminating Chlorophyll: Innovative Chemistry Popularization Experiment. University Chemistry, 2024, 39(9): 359-368. doi: 10.3866/PKU.DXHX202309059
7. [7]
  Zehua Zhang , Haitao Yu , Yanyu Qi . Design Strategy for Thermally Activated Delayed Fluorescence Materials with Multiple Resonance Effect. Acta Physico-Chimica Sinica, 2025, 41(1): 100006-0. doi: 10.3866/PKU.WHXB202309042
8. [8]
  Wenwei Zeng , Qingyu Sun , Mengxiang Liang , Lirong Lin , Laiying Zhang . Unveiling Anti-Counterfeiting Secrets: Excitation-Dependent Luminescence in Sb³⁺-Doped Perovskite Materials. University Chemistry, 2026, 41(2): 375-384. doi: 10.12461/PKU.DXHX202503036
9. [9]
  Zhifeng CAI , Ying WU , Yanan LI , Guiyu MENG , Tianyu MIAO , Yihao ZHANG . Effective detection of malachite green by folic acid stabilized silver nanoclusters. Chinese Journal of Inorganic Chemistry, 2025, 41(5): 983-993. doi: 10.11862/CJIC.20240394
10. [10]
  Lin LI , Le CHEN , Lingjie HOU , Jiaqi JING , Jiayu DING , Tao ZHOU , Ruiping ZHANG . Smartphone-assisted fluorescent silver nanoclusters as ratiometric sensor for visual colorimetric detection of sulfide. Chinese Journal of Inorganic Chemistry, 2025, 41(11): 2261-2271. doi: 10.11862/CJIC.20250130
11. [11]
  Yu Dai , Xueting Sun , Haoyu Wu , Naizhu Li , Guoe Cheng , Xiaojin Zhang , Fan Xia . Determination of the Michaelis Constant for Gold Nanozyme-Catalyzed Decomposition of Hydrogen Peroxide. University Chemistry, 2025, 40(5): 351-356. doi: 10.12461/PKU.DXHX202407052
12. [12]
  Xingyuan Lu , Yutao Yao , Junjing Gu , Peifeng Su . Energy Decomposition Analysis and Its Application in the Many-Body Effect of Water Clusters. University Chemistry, 2025, 40(3): 100-107. doi: 10.12461/PKU.DXHX202405074
13. [13]
  Shuwen SUN , Gaofeng WANG . Two cadmium coordination polymers constructed by varying Ⅴ-shaped co-ligands: Syntheses, structures, and fluorescence properties. Chinese Journal of Inorganic Chemistry, 2024, 40(3): 613-620. doi: 10.11862/CJIC.20230368
14. [14]
  Dongdong YANG , Jianhua XUE , Yuanyu YANG , Meixia WU , Yujia BAI , Zongxuan WANG , Qi MA . Design and synthesis of two coordination polymers for the rapid detection of ciprofloxacin based on triphenylpolycarboxylic acid ligands. Chinese Journal of Inorganic Chemistry, 2024, 40(12): 2466-2474. doi: 10.11862/CJIC.20240266
15. [15]
  Jiming XI , Yukang TENG , Rui ZHANG , Zhenzhong LU . Fluorescent coordination polymers based on anthracene-and pyrene-derivative ligands. Chinese Journal of Inorganic Chemistry, 2025, 41(5): 847-854. doi: 10.11862/CJIC.20240367
16. [16]
  Yuanyu YANG , Jianhua XUE , Yujia BAI , Lulu CUI , Dongdong YANG , Qi MA . Design, synthesis, and detection of Al³⁺ of two zinc complexes based on Schiff base ligands. Chinese Journal of Inorganic Chemistry, 2025, 41(6): 1207-1216. doi: 10.11862/CJIC.20250005
17. [17]
  Yanfen PENG , Xinyue WANG , Tianbao LIU , Xiaoshuo WU , Yujing WEI . Syntheses and luminescence of four Cd(Ⅱ)/Zn(Ⅱ) complexes constructed by 1,3‐bis(4H‐1,2,4‐triazole)benzene. Chinese Journal of Inorganic Chemistry, 2025, 41(7): 1416-1426. doi: 10.11862/CJIC.20250018
18. [18]
  Youbo HU , Donggang LI , Changhua SUN , Zhenzhong LU , Songjun GU . Coordination polymers based on anthracene- and pyrene-derived ligands: Crystal structure, fluorescent property, and framework isomerization. Chinese Journal of Inorganic Chemistry, 2025, 41(8): 1681-1688. doi: 10.11862/CJIC.20250004
19. [19]
  Gaofeng WANG , Shuwen SUN , Lixin Meng , Dequn PENG . Syntheses and fluorescent sensing properties of two coordination polymers based on 9, 9′-dihexyl-2, 7-di(pyridin-4-yl)fluorene. Chinese Journal of Inorganic Chemistry, 2026, 42(2): 331-339. doi: 10.11862/CJIC.20250260
20. [20]
  Zijuan LI , Xuan LÜ , Jiaojiao CHEN , Haiyang ZHAO , Shuo SUN , Zhiwu ZHANG , Jianlong ZHANG , Yanling MA , Jie LI , Zixian FENG , Jiahui LIU . Synthesis of visual fluorescence emission CdSe nanocrystals based on ligand regulation. Chinese Journal of Inorganic Chemistry, 2025, 41(2): 308-320. doi: 10.11862/CJIC.20240138

Get Citation

PDF

XML

Metrics

PDF Downloads(12)
Abstract views(1404)
HTML views(227)

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142