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Amperometric biosensor for total monoamines using a glassy carbon paste electrode modified with human monoamine oxidase B and manganese dioxide particles

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Abstract

We have prepared a biosensor for the determination of the total monoamine content in complex matrices by immobilizing a human monoamine oxidase B (hMAO B) on a glassy carbon paste electrode and adding manganese dioxide microparticles as the mediator. The enzyme hMAO B (expressed in Pichia pastoris and immobilized by using a dialysis membrane) catalyzes the oxidative deamination of monoamines, and this results in the formation of the corresponding aldehyde, ammonia and hydrogen peroxide. The latter was detected at pH 7.5 at a working voltage of 400 mV (vs. Ag/AgCl) by differential pulse voltammetry and amperometrically by applying flow injection analysis. Analytical parameters were established by using phenylethylamine (PEA) as a standard substrate. Peak height and concentration of PEA are linearly related in the 0.5 to 150 μg mL−1 concentration range, and the limits of detection and of quantification are 0.15 and 0.5 μg mL−1 of PEA, respectively. Substrate specificity was investigated with different monoamines including PEA, serotonin, benzylamine, dopamine, tyramine, and norepinephrine. The applicability of the biosensor was successfully tested in a commercial fish sauce that served as a complex matrix. The total monoamine content was calculated as PEA-equivalents.

A novel electrochemical human monoamine oxidase B modified biosensor for the quantification of monoamines was developed. It is based on a glassy carbon paste electrode with manganese dioxide as mediator. The biosensor was characterised and validated and its application was tested in complex matrix.

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Acknowledgments

The authors thank Ms. Milagros Aldeco for her contribution regarding the production and for providing hMAO B.

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Authors and Affiliations

  1. Institute of Pharmaceutical Sciences, Department of Pharmaceutical Chemistry, University of Graz, Schubertstraße 1, 8010, Graz, Austria

    Maximilian Aigner, Dietlind Telsnig, Christian Teubl & Astrid Ortner

  2. Institute of Chemistry, Department of Analytical Chemistry, University of Graz, Universitätsplatz 1, 8010, Graz, Austria

    Kurt Kalcher

  3. Institute of Biochemistry, Graz University of Technology, Petersgasse 10-12/II, 8010, Graz, Austria

    Peter Macheroux & Silvia Wallner

  4. Department of Biochemistry, Emory University Atlanta, 1510 Clifton Rd. NE, Atlanta, GA, 30322, USA

    Dale Edmondson

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  1. Maximilian Aigner
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  2. Dietlind Telsnig
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  3. Kurt Kalcher
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  4. Christian Teubl
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  5. Peter Macheroux
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  6. Silvia Wallner
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  7. Dale Edmondson
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  8. Astrid Ortner
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Correspondence to Astrid Ortner.

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Aigner, M., Telsnig, D., Kalcher, K. et al. Amperometric biosensor for total monoamines using a glassy carbon paste electrode modified with human monoamine oxidase B and manganese dioxide particles. Microchim Acta 182, 925–931 (2015). https://doi.org/10.1007/s00604-014-1404-5

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  • DOI: https://doi.org/10.1007/s00604-014-1404-5

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