Food and Bioprocess Technology

, Volume 10, Issue 10, pp 1834–1843 | Cite as

A New Label-Free Impedimetric Affinity Sensor Based on Cholinesterases for Detection of Organophosphorous and Carbamic Pesticides in Food Samples: Impedimetric Versus Amperometric Detection

  • Francesca Malvano
  • Donatella AlbaneseEmail author
  • Roberto Pilloton
  • Marisa Di Matteo
  • Alessio Crescitelli
Original Paper


Due to their increasing use in agriculture, the presence of pesticide residues in food and water currently represents one of the major issues for the food safety. Among the pesticides, organophosphate and carbamate species are the most used, and their toxicity is mainly due to their inhibitory effect on acetylcholinesterase (AChE). For this reason, a monoenzymatic acetylcholinesterase impedimetric biosensor was developed in order to sensitively detect carbamate and organophosphate compounds with a very fast response. The working principle of the AChE biosensor exploits the capability of carbamate and organophosphate pesticides to form a stable complex with the enzyme, which causes an impedimetric change. The impedimetric biosensor showed a linearity between 5 and 170 ppb for carbamates and 2.5–170 ppb for organophosphate compounds, with a reproducibility (RSD%) interelectrode equal to 4.8 and 3.1% for organophosphates and carbamates, respectively. Moreover, the common amperometric evaluation of AChE inhibition degree was correlated to the impedimetric changes of the electrode surface, showing a good correlation (R 2 = 0.99 for carbamates and R 2 = 0.98 for organophosphates) between the two methods. In contrast to amperometric evaluation that needs a response time of 20 min, impedimetric detection requires only 4 min. Finally, the impedimetric biosensor was used to measure carbaryl and dichlorvos spiked in different concentrations in tap water and lettuce samples, showing a recovery near to 100% for all concentrations and for both pesticides.


Pesticides Electrochemical impedance spectroscopy Affinity biosensors Acetylcholinesterase 

Supplementary material

11947_2017_1955_MOESM1_ESM.docx (1.1 mb)
ESM 1 (DOCX 1107 kb)


  1. Albanese, D., Di Matteo, M., & Pilloton, R. (2012). Quantitative screening and resolution of carbamic and organoposphate pesticides mixture in extra virgin olive oil by acetylcholinesterase-choline oxidase sensor. J Environ Sci Eng A, 1, 68–77.Google Scholar
  2. Andreescu, S., & Marty, J. L. (2006). Twenty years research in cholinesterase biosensors: from basic research to practical applications. Biomolecular Engineering, 23, 1–15.CrossRefGoogle Scholar
  3. Andreescu, S., Avramescu, A., Bala, C., Magear, V., & Marty, J. L. (2002). Detection of organophosphorus insecticides with immobilized acetylcholinesterase—comparative study of two enzyme sensors. Analytical and Bioanalytical Chemistry, 374, 39–45.CrossRefGoogle Scholar
  4. Arduini, F., Ricci, F., Tuta, C. S., Moscone, D., Amine, A., & Palleschi, G. (2006). Detection of carbamic and organophosphorus pesticides in water samples using cholinesterase biosensor based on Prussian Blue modified screen printed electrode. Analytica Chimica Acta, 580, 155–162.CrossRefGoogle Scholar
  5. Arduini, F., Guidone, S., Amine, A., Palleschi, G., & Moscone, D. (2013). Acetylcholinesterase biosensor based on self-assembled monolayer-modified gold-screen printed electrodes for organophosphorus insecticide detection. Sensors and Actuators B: Chemical, 179, 201–208.CrossRefGoogle Scholar
  6. Bahadir, E. B., & Sezginturk, M. K. (2016). A review on impedimetric biosensors. Artif Cells Nanomedicine Biotechnol, 44(1), 248–262.CrossRefGoogle Scholar
  7. Caetano, J., & Machado, A. S. (2008). Determination of carbaryl in tomato “in natura” using an amperometric biosensor based on the inhibition of acetylcholinesterase activity. Sensors and Actuators B, 129, 40–46.CrossRefGoogle Scholar
  8. Chauhan, N., & Pundir, C. S. (2011). An amperometric biosensor based on acetylcholinesterase immobilized onto iron oxide nanoparticles/multi-walled carbon nanotubes modified gold electrode for measurement of organophosphorus insecticides. Analytica Chimica Acta, 701, 66–74.CrossRefGoogle Scholar
  9. Colovic, M. B., Krstic, D. Z., Lazarevic-Pasti, T. D., Bondzic, A. M., & Vasic, V. M. (2013). Acetylcholinesterase inhibitors: pharmacology and toxicology. Current Neuropharmacology, 11, 315–335.CrossRefGoogle Scholar
  10. Darvesh, S., Darvesh, K. V., McDonald, R. S., Mataija, D., Walash, R., Mothana, S., Lockridge, O., & Martin, E. (2008). Carbamates with differential mechanism of inhibition toward acetylcholinesterase and butyrylcholinesterase. Journal of Medicinal Chemistry, 51, 4200–4212.CrossRefGoogle Scholar
  11. Fukuto, T. R. (1990). Mechanism of action of organophosphorus and carbamate insecticides. Environmental Health Perspectives, 87, 245–254.CrossRefGoogle Scholar
  12. Guan, J. G., Miao, Y. Q., & Zhang, Q. J. (2004). Impedimetric biosensors. Journal of Bioscience and Bioengineering, 97(4), 219–226.CrossRefGoogle Scholar
  13. Krstić, D. Z., Colovic, M., Kralj, M. B., Franko, M., Krinulovic, K., Trebse, P., & Vasic, V. (2008). Inhibition of AChE by malathion and some structurally similar compounds. Journal of Enzyme Inhibition and Medicinal Chemistry, 23, 562–573.CrossRefGoogle Scholar
  14. Liu, G., & Lin, Y. (2006). Biosensor based on self-assembling acetylcholinesterase on carbon nanotubes for flow injection/amperometric detection of organophosphate pesticides and nerve agents. Analytical Chemistry, 78, 835–843.CrossRefGoogle Scholar
  15. Malvano, F., ese, D., Pilloton, R., & Di Matteo, M. (2016). A highly sensitive impedimetric label free immunosensor for ochratoxin measurement in cocoa beans. Food Chemistry, 212, 688–694.CrossRefGoogle Scholar
  16. Mehta, J., Vinayak, P., Tuteja, S. K., Chhabra, V. A., Bhardwaj, N., Paul, A. K., Kim, K. H., & Deep, A. (2016). Biosensors and Bioelectronics, 83, 339–346.CrossRefGoogle Scholar
  17. Moscone, D., Volpe, G., Arduini, F., & Micheli, L. (2016). Rapid electrochemical screening methods for food safety and quality. Acta Imeko, 5, 45–50.CrossRefGoogle Scholar
  18. Sanllorente-Méndez, S., Domínguez-Renedo, O., & Arcos-Martínez, J. (2010). Immobilization of acetylcholinesterase on screen-printed electrodes. Application to the determination of arsenic(III). Sensors, 10, 2119–2128.CrossRefGoogle Scholar
  19. Sassolas, A., Prieto-Simon, B., & Marty, J. L. (2012). Biosensors for pesticide detection: new trends. American Journal of Analytical Chemistry, 3, 210–232.CrossRefGoogle Scholar
  20. Storm, J. E., Rozman, K. K., & Doull, J. (2000). Occupational exposure limits for 30 organophosphate pesticides based on inhibition of red blood cell acetylcholinesterase. Toxicology, 150, 1–29.CrossRefGoogle Scholar
  21. Vakurov, A., Simpson, C. E., Daly, C. L., Gibson, T. D., & Millner, P. A. (2004). Acetylcholinesterase-based biosensor electrodes for organophosphate pesticide detection: I. Modification of carbon surface for immobilization of acetylcholinesterase. Biosensors and Bioelectronics, 20(6), 1118–1125.Google Scholar
  22. Valdes -Ramirez, G., Cortina, M., Ramirez Silva, M. T., & Marty, J. L. (2008). Acetylcholinesterase-based biosensors for quantification of carbofuran, carbaryl, methylparaoxon, and dichlorvos in 5% acetonitrile. Analytical and Bioanalytical Chemistry, 392, 699–707.CrossRefGoogle Scholar
  23. Xavier, M. P., Vallejo, B., Marazuela, M. D., Moreno-Bondi, M. C., Baldini, F., & Falai, A. (2000). Fiber optic monitoring of carbamate pesticides using porous glass with covalently bound chlorophenol red. Biosensors and Bioelectronics, 14, 895–905.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2017

Authors and Affiliations

  1. 1.Department of Industrial EngineeringUniversity of SalernoFiscianoItaly
  2. 2.Institute of Atmospheric Pollution Research of the National Council of Research (CNR)RomeItaly
  3. 3.Institute for Microelectronics and Microsystems of the National Council of Research (CNR)NaplesItaly

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