Amperometric H2O2 biosensor based on poly-thionine nanowire/HRP/nano-Au-modified glassy carbon electrode

doi:10.1016/j.snb.2007.09.062

Sensors and Actuators B: Chemical

Volume 129, Issue 2, 22 February 2008, Pages 779-783

https://doi.org/10.1016/j.snb.2007.09.062 Get rights and content

Abstract

Poly-thionine nanowire/HRP/nano-Au composite nanomaterial was developed to fabricate the H₂O₂ biosensor. Poly-thionine nanowires (PTHNWs) have been synthesized by electrodeposition in porous anodic aluminum oxide (AAO) template. The morphological characterization of PTHNWs was examined by scanning electron microscopy (SEM) and transmission electron microscope (TEM). The nanowires prepared by this method can be used to encapsulate horseradish peroxidase (HRP) and nano-Au by in situ electrochemical copolymerization. The resulting PTHNWs–HRP–nano-Au material facilitates electron-transfer process in electrochemical sensor design. The PTHNWs–HRP–nano-Au film-modified electrode showed to be excellent amperometric sensors for H₂O₂. In pH 6.98 phosphate buffer, almost interference-free determination of H₂O₂ is realized at −0.1 V versus SCE with a linear range of 5 × 10⁻⁷ to 1.3 × 10⁻² M, a correlation coefficient of 0.998 and response time <5 s. The sensitivity of the H₂O₂ biosensor is up to 168 μA mM⁻¹ cm⁻² and the detection limit is 0.3 μM. Furthermore, the biosensor exhibited long-term stability, and good reproducibility.

Introduction

Since the discovery of carbon nanotubes by Iijima in 1991 [1], tremendous attention has been paid to one-dimensional nanostructures as nanotubes and nanowires [2], [3], [4]. Nanostructured materials have attracted considerable attention because of their novel optical, electrical, catalytic, and magnetic properties and their potential applications in nanoelectronic devices, nanosensors, catalysts, and information storage systems [5], [6], [7], [8], [9]. Nanowires, by definition, are anisotropic nanocrystals with large length/diameter ratios. Generally, they would have diameters of 1–200 nm and lengths up to several tens of micrometers [10]. Nanowires play an important role as both interconnects and active components in fabricating nanoscale electronic and photonic devices [11]. Various potential applications of nanowires have propelled extensive research on the synthesis of high-density and well-aligned nanowires via different techniques. In the last decade, many methods have been developed for the synthesis of nanowires. Among them, template synthetic method has attracted a good deal of research interest. At present, ordered porous anodic aluminum oxide (AAO) membrane is widely used to prepare nanowires due to its remarkable hardness, uniform pore size, high pore density together with its potentially low-cost and relative ease of their preparation [12]. In the case of nanowire, AAO template-based eletrodeposition seems to be the most efficient way [13].

Thionine (TH) is a phenothiazine redox dye which can be easily dissolved in water and ethanol [14]. The chemical structure of TH is a small planar molecule with two –NH₂ groups symmetrically distributed on each side [15]. Both thionine monomer and the electrogenerated poly-thionine (PTH) have excellent electrocatalytic activity toward the redox of small molecular compounds. Thionine has been used in many sensors, such as H₂O₂ biosensor [16], electrochemical sensor for immunoassay of carcinoembryonic antigen [17], and NADH biosensor [18]. TH is a perfect electronic mediator for the enzymatic electrodes and to our best knowledge, there is no report up to now about synthesis of thionine nanowires. PTHNWs combine the properties of nanowires and the electrocatalytic activity of PTH, as a result, PTHNWs have potential applications in sensor devices.

At present, one challenge in the nanotechnology area is the fabrication and interconnection of enzymes with nanostructured materials and various approaches have been explored toward the conjugation [19], [20]. The adsorption of enzymes onto the nanostructures has been reported, as these materials provide large surface area for enzyme loading and friendly microenvironment to stabilize the immobilized enzymes, which enable further practical applications [21], [22], [23].

In this paper, we synthesized PTHNWs by cyclic voltammetric electrodeposition in AAO template. HRP and nano-Au were encapsulated in situ in PTHNWs (denoted as PTHNWs–HRP–nano-Au) by electrochemical copolymerization for potential biosensor applications. The electrochemical properties of PTHNWs were investigated. The PTHNWs–HRP–nano-Au film-modified GC electrode showed to be excellent amperometric sensors for H₂O₂ at −0.1 V.

Section snippets

Apparatus and reagents

Cyclic voltammetric and amperometric measurements were carried out on CHI 760B electrochemical workstation (Shanghai, China). Scanning electron microscopy (SEM) analysis was performed using a JSM-5600LV microscope (JEOL Ltd., Japan). Transmission electron microscope (TEM) image was taken with a JEM-3010 transmission electron microscope (JEOL Co. Ltd., Japan). A three-electrode cell (10 mL) was employed with the modified glassy carbon (GC) electrode as the working electrode, a saturated calomel

Measurement principle

The mechanism of the H₂O₂ biosensor is based on the following reactions [26]:H₂O₂ + HRP → HRP-I + H₂OHRP-I + THH → HRP-II + TH → HRP + TH⁺TH⁺ + H⁺ + 2e → THH

The hydrogen peroxide diffuses rapidly and is reduced in the presence of HRP in reaction (1); then HRP-I is reduced to HRP by THH (THH and TH⁺ represent reduced and oxidized forms of thionine, respectively), and oxidized TH⁺ can be reduced to THH at the electrode, bringing about a reduction current. The cathodic current is proportional to H₂O₂ concentration in

Conclusion

In this paper, we demonstrated a new way for the fabrication of amperometric H₂O₂ biosensors based on in situ electropolymerization of PTHNWs–HRP–nano-Au with the help of the AAO template. The synthesis procedure of nanowires is simple and low-cost. The experimental results clearly demonstrate that the proposed biosensor allows fast determination of H₂O₂ with high sensitivity, expanded linear response range and good long-term stability. In addition, the immobilization matrix in this work not

Acknowledgements

This work was supported by the NNSF of China (Nos. 20435010, 20375012, 20675028 and 20205005).

Ai Wu Shi graduated from Suzhou University in 2005 majoring in chemistry. She is currently a graduate student of Hunan University.

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Ai Wu Shi graduated from Suzhou University in 2005 majoring in chemistry. She is currently a graduate student of Hunan University.

Feng Li Qu is currently a PhD student of Hunan University majoring in chemistry and the research interests cover chemical and biosensors.

Ming Hui Yang received the PhD degree from College of Chemistry and Chemical Engineering of Hunan University in 2006.

Guo Li Shen is the professor of chemistry, College of Chemistry and Chemical Engineering, Hunan University, Changsha, China. He is also the vice-chairman of the Chemical Sensor Subcommittee of Chinese Analytical Instrumentation Society and acting deputy editor-in-chief of ‘Chemical Sensor’. He graduated from Department of Chemistry, Fudan University, Shanghai in 1961. His research interests cover chemical sensors and biosensors.

Ru Qin Yu is the professor of chemistry, College of Chemistry and Chemical Engineering, Hunan University, Changsha, China. He is also the Member of Chinese Academy of Sciences since 1991 and editor-in-chief of ‘Chemical Sensor’, editorial adviser of Analytical Chimica Acta (Elsevier) and Journal of Chemometrics (Wiley). He graduated from Department of Chemistry, St. Petersburg University, Russia in 1959. His research interests cover chemical sensors and chemometrics.

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Amperometric H2O2 biosensor based on poly-thionine nanowire/HRP/nano-Au-modified glassy carbon electrode

Abstract

Introduction

Section snippets

Apparatus and reagents

Measurement principle

Conclusion

Acknowledgements

Sens. Actuator B

Electrochim. Acta

Chem. Phys. Lett.

Electrochem. Commun.

Carbon

Electrochim. Acta

Cancer Detect. Prev.

Biosens. Bioelectron.

Anal. Biochem.

J. Solid State Chem.

Surf. Sci.

Electrochem. Commun.

J. Biochem. Biophys. Methods.

Biosens. Bioelectron.

Helical microtubules of graphitic carbon

Nature (Lond.)

Crystalline boron nanowires

J. Am. Chem. Soc.

Complementary detection of prostate-specific antigen using In2O3 nanowires and carbon nanotubes

J. Am. Chem. Soc.

Amperometric H₂O₂ biosensor based on poly-thionine nanowire/HRP/nano-Au-modified glassy carbon electrode

Complementary detection of prostate-specific antigen using In₂O₃ nanowires and carbon nanotubes