An amperometric biosensor based on acetylcholinesterase (AChE) immobilized in gelatin was used to develop an assay for the organophosphate paraoxon. The more traditional manner employing preincubation was used for comparison between measurement procedures, although the aim of the study was to examine the performance of the biosensor for real time monitoring of organophosphates. The biosensor was immersed in a reaction chamber and paraoxon was injected inside. We were able to detect 200 pg of paraoxon within one minute or 2.5 ppb when the biosensor was preincubed in the sample solution for 15 minutes. The practical impact and expectations are discussed.

Center of Advanced Studies Faculty of Military Health Sciences University of Defence Hradec Kralove Czech Republic

Center of Biological Defense Techonin Czech Republic

Department of Toxicology Faculty of Military Health Sciences University of Defence Hradec Kralove Czech Republic

Zobrazit více v PubMed

Kuca K., Jun D., Musilek K. Structural requirements of acetylcholinesterase reactivators. Mini Rev. Med. Chem. 2006;6:269–277. PubMed

Kuca K., Cabal J., Kassa J. In vitro reactivation of sarin-inhibited brain acetylchoinesterase from different species by various oximes. J. Enz. Inhib. Med. Chem. 2005;20:227–232. PubMed

Kuca K., Jun D., Bajgar J. Currently used cholinesterase reactivators against nerve agent intoxication: Comparison of their effectivity in vitro. Drug Chem. Toxicol. 2007;30:31–40. PubMed

Pohanka M., Jun D., Kuca K. Photometric microplate assay for estimation of paraoxon inhibited acetylcholinesterase reactivation efficacy. J. Enz. Inhib. Med. Chem. doi: 10.1080/14756360701811023. PubMed DOI

Arduini F., Ricci F., Bourais L., Amine A., Moscone D., Palleschi G. Extraction and detection of pesticides by cholinesterase inhibition in a two-phase system: a strategy to avoid heavy metal interference. Anal. Lett. 2005;38:1703–1719.

No H.Y., Kim Y.A., Lee Y.T., Lee H.S. Cholinesterase-based dipstick assay for the detection of organophosphate and carbamate pesticides. Anal. Chim. Acta. 2007;594:37–43. PubMed

Kim B.M., Abd El-Aty A.M., Hwang T.E., Jin L.T., Kim Y.S., Shim J.H. Development of an acetylcholinesterase-based detection kit for the determination of organophosphorus and carbamate pesticide residues in agricultural samples. Bull. Kor. Chem. Soc. 2007;28:929–935.

Skladal P. Biosensors based on cholinesterase for detection of pesticides. Food Technol. Biotechnol. 1996;34:43–49.

Renault N.J. New trends in biosensors for organophosphorus pesticides. Sensors. 2001;1:60–74.

Okazaki S., Nakagawa H., Fukuda K., Asakura S., Kiuchi H., Shigemori T., Takahashi S. Reactivation of an amperometric organophosphate pesticide biosensor by 2-pyridinealdoxime methochloride. Sens. Actuat. B. 2000;66:131–134.

Longobardi F., Solfrizzo M., Compagnone D., Del Carlo M., Visconti A. Use of electrochemical biosensor and gas chromatography for determination of dichlorvos in wheat. J. Agric. Food Chem. 2005;53:9389–9394. PubMed

Gulla K.C., Gouda M.D., Thakur M.S., Karanth N.G. Reactivation of immobilized acetyl choilnesterase in an amperometric biosensor for organophosphorus pesticide. Biochim. Biophys. Acta. 2002;1597:133–139. PubMed

Del Carlo M., Pepe A., De Gregorio M., Mascini M., Marty J.L., Fournier D., Visconti A., Compagnone D. An electrochemical bioassay for dichlorvos analysis in durum wheat samples. J. Food Prot. 2006;69:1406–1411. PubMed

Skládal P., Krejčí J. Performance of the amperometricbiosensor with immobilized butyrylcholinesterase in organic solvents. Collect. Czech Chem. Commun. 1996;61:985–991.

Pohanka M., Jun D., Kuca K. Amperometric biosensor for evaluation of competitive cholinesterase inhibition by the reactivator HI-6. Anal. Lett. 2007;40:2351–2359.

Ciucu A., Ciucu C. Organic phase amperometric biosensor for detection of pesticides. Roum. Biotechnol. Lett. 2002;7:667–676.

Suprun E., Evtugyn G., Budnikov H., Ricci F., Moscone D., Palleschi G. Acetylchoinesterase sensor based on screen-printed carbon electrode modified with prussian blue. Anal. Bioanal. Chem. 2005;383:597–604. PubMed

Sotiropoulou S., Chaniotakis N.A. Lowering the detection limit of the acetylcholinesterase biosensor using a nanoporous carbon matrix. Anal. Chim. Acta. 2005;530:199–204.

Timur S., Telefoncu A. Actylcholinesterase (AChE) electrodes based on gelatin and chitosan matrices for the pesticide detection. Artif. Cells Blood Substit. Immobil. Biotechnol. 2004;32:427–442. PubMed

Andreescu S., Marty J.L. Twenty years research in cholinesterase biosensors: from basic research to practical applications. Biomol. Eng. 2006;23:1–15. PubMed

Pohanka M., Kuca K., Jun D. Amperometric biosensor for pesticide methamidophos assay. Acta Medica. 2007;50:239–241. PubMed

Pohanka M., Treml F., Hubálek M., Banďouchová H., Beklová M., Pikula J. Piezoelectric biosensor for a simple serological diagnosis of tularemia in infected european brown hares (Lepus europaeus) Sensors. 2007;7:2825–2834. PubMed PMC

Pohanka M., Pavlis O., Skládal P. Rapid characterization of monoclonal antibodies using the piezoelectric immunosensor. Sensors. 2007;7:341–353.

Pohanka M., Pavliš O., Skládal P. Diagnosis of tularemia using piezoelectric biosensor technology. Talanta. 2007;71:981–985. PubMed

Vitecek J., Petrlova J., Adam V., Havel L., Kramer K.J., Babula P., Kizek R. A fluorimetric sensor for detection of one living cell. Sensors. 2007;7:222–238.

Pohanka M., Skládal P. Piezoelectric immunosensor for the direct and rapid detection of Francisella tularensis. Folia Microbiol. 2007;52:325–330. PubMed

Pohanka M., Skládal P., Pavliš O. A label free piezoelectric immunosensor for rapid assay of Escherichia coli. J. Immunoass. Immunochem. 2008;29:70–79. PubMed

Pohanka M., Kuca K., Jun D. Aflatoxin assay using an amperometric sensor strip and acetylcholinesterase as recognition element. Sens. Lett. 2008;6:450–453.

Pohanka M., Jun D., Kalasz H., Kuca K. Cholinesterase biosensor construction – a review. Protein Peptide Lett. 2008;15:795–798. PubMed

Variation of cholinesterase-based biosensor sensitivity to inhibition by organophosphate due to ionizing radiation

Cholinesterase based amperometric biosensors for assay of anticholinergic compounds

Evaluation of cholinesterase activities during in vivo intoxication using an electrochemical sensor strip - correlation with intoxication symptoms