Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

A novel fluorescent “turn-on” aptasensor based on nitrogen-doped graphene quantum dots and hexagonal cobalt oxyhydroxide nanoflakes to detect tetracycline


In this study, a novel fluorescent “turn-on” aptasensor was developed for sensitive and rapid detection of tetracycline (TC) in animal-derived food. It is based on aptamer-functionalized nitrogen-doped graphene quantum dots (N-GQDs-aptamer) coupled with cobalt oxyhydroxide (CoOOH) nanoflakes. The CoOOH nanoflakes are efficient fluorescence quenchers in homogeneous solutions, and this is due to their advantages of excellent optical properties, superior flexibility, and water dispersibility. The proposed method’s mechanism is driven by quenching based on the fluorescence resonance energy transfer (FRET) between the donor (N-GQDs) and the acceptor (CoOOH nanoflakes). On the other hand, fluorescence recovery is caused by the structure switching behavior of the aptamer. Compared with previous methods, our developed method exhibits better behavior in terms of being easy to fabricate and being simple in detection procedure and maintains the detection limit low enough in TC determination: a linear range from 1 to 100 ng mL−1 and a detection limit of 0.95 ng mL−1 (S/N = 3). Furthermore, the proposed method was applied to five animal-derived food samples (milk, honey, fish, eggs, and chicken muscle) and demonstrated practical applicability. As well, the method has the advantages of simplicity in pre-treatment and convenience in instruments, saves times, and is cost-effective. Finally, the proposed method demonstrates significant potential for sensitive and rapid detection of specific components in real samples.

Graphical abstract

This is a preview of subscription content, log in to check access.

Scheme 1
Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5


  1. 1.

    Zhang L, Chen L. Fluorescence probe based on hybrid mesoporous silica/quantum dot/molecularly imprinted polymer for detection of tetracycline. ACS Appl Mater Interfaces. 2016;8(25):16248–56.

  2. 2.

    Bougrini M, Florea A, Cristea C, Sandulescu R, Vocanson F, Errachid A, et al. Development of a novel sensitive molecularly imprinted polymer sensor based on electropolymerization of a microporous-metal-organic framework for tetracycline detection in honey. Food Control. 2016;59:424–9.

  3. 3.

    Van Boeckel TP, Pires J, Silvester R, Zhao C, Song J, Criscuolo NG, et al. Global trends in antimicrobial resistance in animals in low- and middle-income countries. Science. 2019;365(6459).

  4. 4.

    Feng MX, Wang GN, Yang K, Liu HZ, Wang JP. Molecularly imprinted polymer-high performance liquid chromatography for the determination of tetracycline drugs in animal derived foods. Food Control. 2016;69:171–6.

  5. 5.

    Li H, Chen Q, Mehedi Hassan M, Chen X, Ouyang Q, Guo Z, et al. A magnetite/PMAA nanospheres-targeting SERS aptasensor for tetracycline sensing using mercapto molecules embedded core/shell nanoparticles for signal amplification. Biosens Bioelectron. 2017;92:192–9.

  6. 6.

    Taghdisi SM, Danesh NM, Ramezani M, Abnous K. A novel M-shape electrochemical aptasensor for ultrasensitive detection of tetracyclines. Biosens Bioelectron. 2016;85:509–14.

  7. 7.

    Chen JL. Determination of tetracycline using imprinted polymethacrylates along with fluorescent CdTe quantum dots on plastic substrates. Microchim Acta. 2017;184(5):1335–43.

  8. 8.

    Jiang Y, Shi M, Liu Y, Wan S, Cui C, Zhang L, et al. Aptamer/AuNP biosensor for colorimetric profiling of exosomal proteins. Angew Chem Int Ed. 2017;56(39):11916–20.

  9. 9.

    Ye S, Ye R, Shi Y, Qiu B, Guo L, Huang D, et al. Highly sensitive aptamer based on electrochemiluminescence biosensor for label-free detection of bisphenol A. Anal Bioanal Chem. 2017;409(30):7145–51.

  10. 10.

    Ma H, Liu J, Ali MM, Mahmood MAI, Labanieh L, Lu M, et al. Nucleic acid aptamers in cancer research, diagnosis and therapy. Chem Soc Rev. 2015;44(5):1240–56.

  11. 11.

    Jalalian SHKN, Ramezani M. Electrochemical and optical aptamer-based sensors for detection of tetracyclines. Trends Food Sci Technol. 2018;73:45–57.

  12. 12.

    Feng C, Dai S, Wang L. Optical aptasensors for quantitative detection of small biomolecules: a review. Biosens Bioelectron. 2014;59:64–74.

  13. 13.

    Zhou C, Zou H, Sun C, Ren D, Xiong W, Li Y. Fluorescent aptasensor for detection of four tetracycline veterinary drugs in milk based on catalytic hairpin assembly reaction and displacement of G-quadruplex. Anal Bioanal Chem. 2018;410(12):2981–9.

  14. 14.

    Kim SLHJ. Gold nanostar enhanced surface plasmon resonance detection of an antibiotic at attomolar concentrations via an aptamer-antibody sandwich assay. Anal Chem. 2017;89(12):6624–30.

  15. 15.

    Das R, Dhiman A, Kapil A, Bansal V, Sharma TK. Aptamer-mediated colorimetric and electrochemical detection of Pseudomonas aeruginosa utilizing peroxidase-mimic activity of gold NanoZyme. Anal Bioanal Chem. 2019;411(6):1229–38.

  16. 16.

    Zhan X, Hu G, Wagberg T, Zhan S, Xu H, Zhou P. Electrochemical aptasensor for tetracycline using a screen-printed carbon electrode modified with an alginate film containing reduced graphene oxide and magnetite (Fe3O4) nanoparticles. Microchim Acta. 2015;183(2):723–9.

  17. 17.

    He Y, Zhang B, Fan Z. Aptamer based fluorometric sulfamethazine assay based on the use of graphene oxide quantum dots. Microchim Acta. 2018;185(3):163.

  18. 18.

    Cen Y, Tang J, Kong X, Wu S, Yuan J, Yu R, et al. A cobalt oxyhydroxide-modified upconversion nanosystem for sensitive fluorescence sensing of ascorbic acid in human plasma. Nanoscale. 2015;7(33):13951–7.

  19. 19.

    Li G, Kong W, Zhao M, Lu S, Gong P, Chen G, et al. A fluorescence resonance energy transfer (FRET) based “turn-on” nanofluorescence sensor using a nitrogen-doped carbon dot-hexagonal cobalt oxyhydroxide nanosheet architecture and application to alpha-glucosidase inhibitor screening. Biosens Bioelectron. 2016;79:728–35.

  20. 20.

    Ji D, Du Y, Meng H, Zhang L, Huang Z, Hu Y, et al. A novel colorimetric strategy for sensitive and rapid sensing of ascorbic acid using cobalt oxyhydroxide nanoflakes and 3,3′,5,5′-tetramethylbenzidine. Sensors Actuators B Chem. 2018;256:512–9.

  21. 21.

    Meng HM, Zhang XB, Yang C, Kuai H, Mao GJ, Gong L, et al. Efficient two-photon fluorescence nanoprobe for turn-on detection and imaging of ascorbic acid in living cells and tissues. Anal Chem. 2016;88(11):6057–63.

  22. 22.

    Lin L, Rong M, Lu S, Song X, Zhong Y, Yan J, et al. A facile synthesis of highly luminescent nitrogen-doped graphene quantum dots for the detection of 2,4,6-trinitrophenol in aqueous solution. Nanoscale. 2015;7(5):1872–8.

  23. 23.

    Zhu L, Xu G, Song Q, Tang T, Wang X, Wei F, et al. Highly sensitive determination of dopamine by a turn-on fluorescent biosensor based on aptamer labeled carbon dots and nano-graphite. Sensors Actuators B Chem. 2016;231:506–12.

  24. 24.

    Li L, Wang C, Liu K, Wang Y, Liu K, Lin Y. Hexagonal cobalt oxyhydroxide-carbon dots hybridized surface: high sensitive fluorescence turn-on probe for monitoring of ascorbic acid in rat brain following brain ischemia. Anal Chem. 2015;87(6):3404–11.

  25. 25.

    Yan X, Song Y, Zhu C, Li H, Du D, Su X, et al. MnO2 nanosheet-carbon dots sensing platform for sensitive detection of organophosphorus pesticides. Anal Chem. 2018;90(4):2618–24.

  26. 26.

    Sun C, Su R, Bie J, Sun H, Qiao S, Ma X, et al. Label-free fluorescent sensor based on aptamer and thiazole orange for the detection of tetracycline. Dyes Pigments. 2018;149:867–75.

  27. 27.

    Hou J, Li H, Wang L, Zhang P, Zhou T, Ding H, et al. Rapid microwave-assisted synthesis of molecularly imprinted polymers on carbon quantum dots for fluorescent sensing of tetracycline in milk. Talanta. 2016;146:34–40.

  28. 28.

    Shen M, Duan N, Wu S, Zou Y, Wang Z. Polydimethylsiloxane gold nanoparticle composite film as structure for aptamer-based detection of Vibrio parahaemolyticus by surface-enhanced Raman spectroscopy. Food Anal Methods. 2018;12(2):595–603.

  29. 29.

    Li Y, Huang H, Ma Y, Tong J. Highly sensitive fluorescent detection of dihydroxybenzene based on graphene quantum dots. Sensors Actuators B Chem. 2014;205:227–33.

  30. 30.

    Zhou X, Wang A, Yu C, Wu S, Shen J. Facile synthesis of molecularly imprinted graphene quantum dots for the determination of dopamine with affinity-adjustable. ACS Appl Mater Interfaces. 2015;7(22):11741–7.

  31. 31.

    Zhou Y, Qu ZB, Zeng Y, Zhou T, Shi G. A novel composite of graphene quantum dots and molecularly imprinted polymer for fluorescent detection of paranitrophenol. Biosens Bioelectron. 2014;52:317–23.

  32. 32.

    Wang Y, Gan N, Zhou Y, Li T, Cao Y, Chen Y. Novel single-stranded DNA binding protein-assisted fluorescence aptamer switch based on FRET for homogeneous detection of antibiotics. Biosens Bioelectron. 2017;87:508–13.

  33. 33.

    Niazi JH, Lee SJ, Kim YS, Gu MB. ssDNA aptamers that selectively bind oxytetracycline. Bioorg Med Chem. 2008;16(3):1254–61.

  34. 34.

    Wang S, Liu J, Yong W, Chen Q, Zhang L, Dong Y, et al. A direct competitive assay-based aptasensor for sensitive determination of tetracycline residue in honey. Talanta. 2015;131:562–9.

  35. 35.

    Yang C, Bie J, Zhang X, Yan C, Li H, Zhang M, et al. A label-free aptasensor for the detection of tetracycline based on the luminescence of SYBR green. Spectrochim Acta A. 2018;202:382–8.

Download references


We would like to thank the National Key R&D Program of China (No. 2016YFD0401202) and the Special Project of Tianjin Innovation Platform (No. 17PTGCCX00230).

Author information

Correspondence to Junping Wang or Shuo Wang.

Ethics declarations

Conflict of interest

The authors declare that they have no competing interests.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material


(PDF 529 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Zhang, L., Wang, J., Deng, J. et al. A novel fluorescent “turn-on” aptasensor based on nitrogen-doped graphene quantum dots and hexagonal cobalt oxyhydroxide nanoflakes to detect tetracycline. Anal Bioanal Chem 412, 1343–1351 (2020).

Download citation


  • Aptamer
  • Cobalt oxyhydroxide nanoflakes
  • Fluorescence resonance energy transfer
  • Graphene quantum dots
  • Structure switching