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An efficient non-mediated amperometric biosensor for nitrite determination.

Célia M Silveira1, Sofia P Gomes, Alberto N Araújo

  • 1REQUIMTE-Dept. de Química, CQFB, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal.

Biosensors & Bioelectronics
|March 2, 2010
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Summary

We developed a novel electrochemical biosensor for nitrite detection using cytochrome c nitrite reductase (ccNiR) immobilized in sol-gel silica. This mediator-free sensor offers high sensitivity and stability for analyzing nitrites in complex environmental samples.

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Area of Science:

  • Electrochemistry
  • Biosensor Technology
  • Environmental Science

Background:

  • Nitrite determination is crucial for environmental monitoring.
  • Previous electrochemical biosensors often rely on redox mediators, complicating their design and application.
  • Cytochrome c nitrite reductase (ccNiR) offers high turnover and electron transfer rates, making it a promising enzyme for biosensing.

Purpose of the Study:

  • To construct a novel, non-mediated electrochemical biosensor for sensitive and selective nitrite determination.
  • To exploit the direct electron transfer capabilities of cytochrome c nitrite reductase (ccNiR).
  • To incorporate ccNiR into a stable matrix for practical application in complex samples.

Main Methods:

  • Construction of a bioelectrode by immobilizing ccNiR from Desulfovibrio desulfuricans within a porous silica glass matrix using the sol-gel process on a pyrolytic graphite (PG) surface.
  • Electrochemical characterization of the ccNiR/sol-gel/PG electrode for nitrite detection.
  • Evaluation of sensor performance including sensitivity, linear range, detection limit, and stability.

Main Results:

  • The ccNiR/sol-gel/PG electrode demonstrated direct electron transfer and catalytic currents in the presence of nitrite, without redox mediators.
  • The biosensor achieved a low minimum detectable concentration of 120 nM nitrite.
  • High sensitivity (430 mA M⁻¹ cm⁻²) was observed within a linear range of 0.25–50 µM, with stable performance for up to two weeks.
  • Accurate nitrite analysis in freshwater samples was achieved using the standard addition method.

Conclusions:

  • A novel, mediator-free electrochemical biosensor for nitrite detection has been successfully developed.
  • The sol-gel immobilization of ccNiR on PG enables direct electron transfer, overcoming limitations of previous designs.
  • The developed biosensor exhibits excellent sensitivity, stability, and accuracy, suitable for environmental monitoring applications.