Elsevier

Electrochimica Acta

Volume 169, 1 July 2015, Pages 326-333
Electrochimica Acta

The use of different glucose oxidases for the development of an amperometric reagentless glucose biosensor based on gold nanoparticles covered by polypyrrole

https://doi.org/10.1016/j.electacta.2015.04.072Get rights and content

ABSTRACT

The amperometric glucose biosensors based on adsorbed electron transfer mediator (ETM) tetrathiafulvalene (TTF) or 1,10-phenanthroline-5,6-dione (PD) and glucose oxidase (GOx) from Aspergillus niger (GOxA.niger), Penicillium adametzii (GOxP.adametzii) or Penicillium funiculosum (GOxP.funiculosum) cross-linked with glutaraldehyde were investigated. ETM and enzyme were immobilized layer by layer on bare graphite rod electrode (GR) premodified with gold nanoparticles (AuNP) of (i) 3.5 nm (GOx/ETM/AuNP3.5/GR), (ii) 6.0 nm (GOx/ETM/AuNP6.0/GR) and (iii) 13.0 nm (GOx/ETM/AuNP13.0/GR) size. The amperometric signals for all the developed biosensors were higher using PD in comparison with TTF. The biosensor based on GOxP.funiculosum showed higher analytical signal to glucose in a comparison to biosensors based on GOxA.niger and GOxP.adametzii. The registered current to glucose using GOxP.funiculosum/PD/AuNP3.5/GR electrode was linear in the glucose range from 0.1 to 10.0 mmol L−1 and the limit of detection was 0.024 mmol L−1. Enzymatical synthesis of polypyrrole (Ppy) layer on the electrode was applied in order to expand the linear glucose detection range. After 22 h of polymerization the amperometric signal was linear in the glucose concentration range from 0.1 to 25.0 mmol L−1, while after 69 h this rage was increased up to 50.0 mmol L−1. Additionally Ppy layer on the electrode surface reduced the influence of interfering species on the amperometric signal. The performance of developed biosensor was investigated in human serum samples.

Section snippets

INTRODUCTION

Glucose biosensor-related research has tremendous interest from the introduction of the first glucose biosensor based on glucose oxidase (GOx) in 1962 [1]. The electrochemical GOx-based biosensors are still among the most widely used, although many improvements have been added since the 1960’s. Currently, these biosensors are applied in different areas such as food and pharmaceutical industry and particularly in clinical diagnostics because diabetes is a worldwide public health problem [2], [3]

Chemicals

GOxA.niger, type VII was purchased from Sigma–Aldrich (Buchs, Switzerland). TTF and PD were purchased from Sigma–Aldrich (Buchs, Switzerland). d-(+)-glucose, tetrachloroauric acid (HAuCl4 × 3H2O) and tannic acid were obtained from Carl Roth GmbH&Co (Karlsruhe, Germany), NaH2PO4 × 2H2O and Na2PO4 × 12H2O – from Reachim (Saint Petersburg, Russia). CH3COONa was purchased from Penta (Praha, Czech Republic), HCl – from Acta Medica (Hradec Kralove, Czech Republic), KCl – from Lachema (Neratovice, Czech

Results

The principle of electrochemical biosensors is usually based on the monitoring of current associated with oxidation or reduction of an electroactive species involved in the recognition process [40]. In our research, redox mediators TTF or PD were adsorbed on bare GR or AuNP/GR electrode surface and then corresponding GOx (from Aspergillus niger, Penicillium adametzii or Penicillium funiculosum) was immobilized onto the physically modified graphite surface by cross-linking with glutaraldehyde

Conclusions

A comparative study of GOx from Aspergillus niger, Penicillium adametzii, Penicillium funiculosum and AuNP of different size for amperometric glucose detection by reagentless biosensor was performed. Graphite electrode was modified with 3.5, 6.0 or 13.0 nm AuNP, TTF or PD as electron transfer mediator and GOx. The biosensor based on GOxP.funiculosum/PD/AuNP3.5/GR electrode was characterized by quick response, high registered current and sensitivity (52.1 μA cm−2 (mmol L−1)−1, linear glucose

Acknowledgement

This research was funded by the European Social Fund under the Global Grant measure. Authors thank the master student Liliana Butko for technical assistance.

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