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

Fabrication and evaluation of a label-free piezoelectric immunosensor for sensitive and selective detection of amantadine in foods of animal origin


A label-free piezoelectric immunosensor was fabricated and applied to the detection of the antiviral drug amantadine (AM) in foods of animal origin. Experimental parameters associated with the fabrication and measurement process were optimized and are discussed here in detail. The proposed piezoelectric sensor is based on an immunosuppression format and uses a portable quartz crystal microbalance (QCM) chip. It was found to provide a good response to AM, with a sensitivity and limit of detection (LOD) of 33.9 and 1.3 ng mL−1, respectively, as well as low cross-reactivity (CR, < 0.01%) with AM analogues. The immunosensor was further applied to quantify AM at three levels in spiked samples of typical foods of animal origin, and yielded recoveries of 83.2–93.4% and standard deviations (SDs, n = 3) of 2.4–4.5%, which are comparable to the results (recoveries: 82.6–94.3%; SDs: 1.7–4.2%) obtained using a high-performance liquid chromatography–tandem mass spectrometry (HPLC-MS/MS) method. Furthermore, the piezoelectric immunosensing chip can be regenerated multiple (at least 20) times with low signal attenuation (about 10%). A sample analysis can be completed within 50 min (sample pretreatment: about 40 min, QCM measurement: 5 min). These results demonstrate that the developed piezoelectric immunosensor provides a sensitive, accurate, portable, and low-cost analytical strategy for the antiviral drug AM in foods of animal origin, and this label-free detection method could also be applied to analyze other targets in the field of food safety.

Graphical abstract

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6


  1. 1.

    Chew CF, Guy A, Biggin PC. Distribution and dynamics of adamantanes in a lipid bilayer. Biophys J. 2008;95:5627–36.

  2. 2.

    Foller M, Geiger C, Mahmud H, Nicolay J, Lang F. Stimulation of suicidal erythrocyte death by amantadine. Eur J Pharmacol. 2008;581:13–8.

  3. 3.

    Wang J, Schnell JR, Chou JJ. Amantadine partition and localization in phospholipid membrane: a solution NMR study. Biochem Biophys Res Commun. 2004;324:212–7.

  4. 4.

    Suwalsky M, Jemiola-Rzeminska M, Altamirano M, Villena F, Dukes N, Strzalka K. Interactions of the antiviral and antiparkinson agent amantadine with lipid membranes and human erythrocytes. Biophys Chem. 2015;202:13–20.

  5. 5.

    He G, Qiao H, Dong C, He C, Zhao L, Tian Y. Amantadine-resistance among H5N1 avian influenza viruses isolated in northern China. Antivir Res. 2008;77:72–6.

  6. 6.

    Nishikawa N, Nagai M, Moritoyo T, Yabe H, Nomoto M. Plasma amantadine concentrations in patients with Parkinson’s disease. Parkinsonism Relat D. 2009;15:351–3.

  7. 7.

    Wu Y, Chen R, Xue Y, Yang T, Zhao J, Zhu Y. Simultaneous determination of amantadine, rimantadine and memantine in chicken muscle using multi-walled carbon nanotubes as a reversed-dispersive solid phase extraction sorbent. J Chromatogr B. 2014;965:197–205.

  8. 8.

    Hu Y, Li J, Zhang Z, Zhang H, Luo L, Yao S. Imprinted sol-gel electrochemical sensor for the determination of benzylpenicillin based on Fe3O4@SiO2/multi-walled carbon nanotubes-chitosans nanocomposite film modified carbon electrode. Anal Chim Acta. 2011;698:61–8.

  9. 9.

    Bardajee GR, Hooshyar Z, Soleyman R. Nanocomposites of sodium alginate biopolymer and CdTe/ZnS quantum dots for fluorescent determination of amantadine. J Polym Res. 2017;24:128.

  10. 10.

    Yeh HH, Yang Y, Chen S. Simultaneous determination of memantine and amantadine in human plasma as fluorescein derivatives by micellar electrokinetic chromatography with laser-induced fluorescence detection and its clinical application. Electrophoresis. 2010;31:1903–11.

  11. 11.

    Farajzadeh MA, Nouri N, Nabil AA. Determination of amantadine in biological fluids using simultaneous derivatization and dispersive liquid-liquid microextraction followed by gas chromatography-flame ionization detection. J Chromatogr B. 2013;940:142–9.

  12. 12.

    Dou Y, Sun Y, Ren Y, Ju P, Ren Y. Simultaneous non-destructive determination of two components of combined paracetamol and amantadine hydrochloride in tablets and powder by NIR spectroscopy and artificial neural networks. J Pharm Biomed Anal. 2005;37:543–9.

  13. 13.

    Yan H, Liu X, Cui F, Yun H, Li J, Ding S, et al. Determination of amantadine and rimantadine in chicken muscle by QuEChERS pretreatment method and UHPLC coupled with LTQ orbitrap mass spectrometry. J Chromatogr B. 2013;938:8–13.

  14. 14.

    Xu L, Peng S, Liu L, Song S, Kuang H, Xu C. Development of sensitive and fast immunoassays for amantadine detection. Food Agr Immunol. 2016;27:678–88.

  15. 15.

    Xie S, Wen K, Xie J, Zheng Y, Peng T, Wang J, et al. Magnetic-assisted biotinylated single-chain variable fragment antibody-based immunoassay for amantadine detection in chicken. Anal Bioanal Chem. 2018;410:6197–205.

  16. 16.

    Pan M, Yang J, Li S, Wen W, Wang J, Ding Y, et al. A reproducible surface plasmon resonance immunochip for the label-free detection of amantadine in animal-derived foods. Food Anal Method. 2019;12:1007–16.

  17. 17.

    Wu S, Zhu F, Hu L, Xia J, Xu G, Liu D, et al. Development of a competitive immunochromatographic assay for the sensitive detection of amantadine in chicken muscle. Food Chem. 2017;232:770–6.

  18. 18.

    Hao X, Li N, Xu Z, Li N, Luo H. An electrochemical sensing strategy for amantadine detection based on competitive host-guest interaction of methylene blue/beta-cyclodextrin/poly(N-acetylaniline) modified electrode. Electroanal. 2016;28:1489–94.

  19. 19.

    Yun Y, Pan M, Fang G, Yang Y, Guo T, Deng J, et al. Molecularly imprinted electrodeposition o-aminothiophenol sensor for selective and sensitive determination of amantadine in animal-derived foods. Sensor Actuat. B-Chem. 2017;238:32–9.

  20. 20.

    Yun Y, Pan M, Fang G, Gu Y, Wen W, Xue R, et al. An electrodeposited molecularly imprinted quartz crystal microbalance sensor sensitized with AuNPs and rGO material for highly selective and sensitive detection of amantadine. RSC Adv. 2018;8:6600–7.

  21. 21.

    Shang Y, Singh PR, Chisti MM, Mernaugh R, Zeng X. Immobilization of a human epidermal growth factor receptor 2 mimotope-derived synthetic peptide on Au and its potential application for detection of herceptin in human serum by quartz crystal microbalance. Anal Chem. 2011;83:8928–36.

  22. 22.

    Kojima T. Combined reflectometric interference spectroscopy and quartz crystal microbalance detect differential adsorption of lipid vesicles with different phase transition temperatures on SiO2, TiO2, and Au surfaces. Anal Chem. 2017;89:13596–602.

  23. 23.

    Niloofar A, Pedram F. Adsorption characteristics of carboxymethylated lignin on rigid and soft surfaces probed by quartz crystal microbalance. Langmuir. 2018;34:15293–303.

  24. 24.

    Da-Silva E, Baudart J, Barthelmebs L. Biosensing platforms for Vibrio bacteria detection based on whole cell and nucleic acid analysis: a review. Talanta. 2018;190:410–22.

  25. 25.

    Chauhan R, Nagar B, Solanki PR, Basu T. Development of triglyceride biosensor based on a platinum nano particle and polypyrrole nano composite electrode. Materials Focus. 2013;2:316–23.

  26. 26.

    Chauhan R, Solanki PR, Singh J, Mukherjee I, Basu T, Malhotra BD. A novel electrochemical piezoelectric label free immunosensor for aflatoxin B1 detection in groundnut. Food Control. 2015;52:60–70.

  27. 27.

    Jaiswal N, Pandey CM, Soni A, Tiwari I, Rosillo-Lopez M, Salzmann CG, et al. Electrochemical genosensor based on carboxylated graphene for detection of water-borne pathogen. Sensor Actuat. B-Chem. 2018;275:312–21.

  28. 28.

    Ding J, Lu Z, Wang R, Shen G, Xiao L. Piezoelectric immunosensor with gold nanoparticles enhanced competitive immunoreaction technique for 2,4-dichlorophenoxyacetic acid quantification. Sensor Actuat. B-Chem. 2014;193:568–73.

  29. 29.

    Li N, Chow AM, Ganesh HVS, Brown IR, Kerman K. Quantum dot based fluorometric detection of cancer TF-antigen. Anal Chem. 2013;85:9699–704.

  30. 30.

    Zong C, Wu J, Wang C, Ju H, Yan F. Chemiluminescence imaging immunoassay of multiple tumor markers for cancer screening. Anal Chem. 2012;84:2410–5.

  31. 31.

    Zhou J, Du L, Zou L, Zou Y, Hu N, Wang P. An ultrasensitive electrochemical immunosensor for carcinoembryonic antigen detection based on staphylococcal protein A-Au nanoparticle modified gold electrode. Sensor Actuat B-Chem. 2014;197:220–7.

  32. 32.

    Zhu Y, Peng J, Jiang L, Zhu J. Fluorescent immunosensor based on CuS nanoparticles for sensitive detection of cancer biomarker. Analyst. 2014;139:649–55.

  33. 33.

    Zhang J, Chen X, Yang M. Enzyme modified peptide nanowire as label for the fabrication of electrochemical immunosensor. Sensor Actuat. B-Chem. 2014;196:189–93.

  34. 34.

    Salmain M, Ghasemi M, Boujday S, Spadavecchia J, Techer C, Val F, et al. Piezoelectric immunosensor for direct and rapid detection of staphylococcal enterotoxin A (SEA) at the ng level. Biosens Bioelectron. 2011;29:140–4.

  35. 35.

    Yan Z, Yang M. WangZ, Zhang F, Xia J, Shi G, Xia L, Li Y, Xia Y, Xia L. A label-free immunosensor for detecting common acute lymphoblastic leukemia antigen (CD10) based on gold nanoparticles by quartz crystal microbalance. Sensor Actuat. B-Chem. 2015;210:248–53.

Download references


This work was financially supported by the National Key R&D Program of China (no. 2017YFE0110800), the EUH2020 (no. EU-China-Safe 727564), the Tianjin Natural Science Foundation (no. 17JCQNJC14800), the National Natural Science Foundation of China (project no. 31301461), Basic Research Fees of Universities and Colleges in Tianjin (no. 2017KDZD01), the Open Project Program of State Key Laboratory of Food Nutrition and Safety (no.SKLFNS-KF-201803), the Project Program of Key Laboratory of Food Nutrition and Safety, Ministry of Education (no. 2018001), and the Foundation (no. GCZX201801) of the Tianjin Engineering Research Center of Safety Control Technology in Food Processing, China.

Author information

Correspondence to Mingfei Pan or Shuo Wang.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note

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

Electronic supplementary material


(PDF 74 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Yun, Y., Pan, M., Wang, L. et al. Fabrication and evaluation of a label-free piezoelectric immunosensor for sensitive and selective detection of amantadine in foods of animal origin. Anal Bioanal Chem 411, 5745–5753 (2019).

Download citation


  • Amantadine
  • Piezoelectric immunosensor
  • Quartz crystal microbalance
  • Label-free