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A novel fabrication approach of a highly sensitive amperometric glucose biosensor based on a single ZnO
nanofiber (ZONF) is presented. Nanofibers (NFs) of poly(vinyl pyrrolidone)/zinc acetate composite have
been synthesized by electrospinning technique. By high-temperature calcinations of the above precursor fibers,
ZONFs with diameters in the range of 350-195 nm have been successfully obtained. A single NF on a gold
electrode is functionalized with glucose oxidase (GOx) by physical adsorption. Electrochemical measurements
of the biosensor revealed a high and reproducible sensitivity of 70.2 A cm-2 mM-1 within a response time
of less than 4 s. The biosensor also showed a linear range from 0.25 to 19 mM with a low limit of detection
(LOD) of 1 M. Furthermore, it has been revealed that the biosensor exhibits a good anti-interference ability
and favorable stability over relatively long-term storage (more than 4 months). All these results strongly
suggest that a single ZONF can provide a new platform for biosensor design and other biological applications.
I. Introduction
The unique and fascinating properties of nanostructured
materials have triggered tremendous motivation among scientists
to explore the possibilities of using them in industrial and
medical applications. Biosensors are becoming essential in the
fields of health care, chemical and biological analysis, environmental monitoring, and good processing industries.1,2 Among
them, glucose sensors, as one of the most popular biosensors,
have been extensively investigated due to their important clinical
applications. The fast and accurate determination of glucose has
profound applications, since glucose concentration is a crucial
indicator in many diseases, such as diabetes and endocrine
metabolic disorder. In recent years, many efforts have been made
to develop reliable glucose biosensors using electrochemical
methods,3 chemiluminescence,4 or other methods.5 Among all
the methods, the enzyme-involved electrochemical glucose
biosensor has been widely studied because of its simplicity, high
selectivity and relative low cost.6-8 Among the numerous reports
in glucose biosensors, the immobilization of enzymes on a
suitable matrix and their stability are important factors in the
fabrication of the biosensors.9 The immobilization of glucose
oxidase (GOx), a widely used analytical enzyme for glucose
detection, has been realized by various methods, such as physical
adsorption, cross-linking, self-assembly, incorporation in carbon
paste, polymers, and sol-gels, etc.10-15
On the other hand, nanostructures have unique advantages
in immobilization enzymes and can retain their bioactivity as a
result of the high surface area for higher enzyme loading,
desirable microenvironment, and the direct electron transfer
between the enzymes active sites and the electrode.16-19
Glucose biosensors, making use of a titania sol-gel membrane,
carbon nanotubes, Au nanoparticles, TiO2 nanoporous film, and
ZrO2/chitasan composite film to immobilize enzymes, have been
reported.15,20-22 Recently, ZnO and its one-dimensional (ID)
nanostructures have been investigated intensively due to their
* Crossponding author E-mail: Jzhu@mail.tsinghua.edu.cn, Fax: 86-1062771160.
Figure 1. (a) Schematic of electrospinning experimental setup used for the fabrication of ZONFs. (b) SEM image of the as-prepared NF. (c) SEM
image of the NF after calcinations at 700 C for 5 h. (d) XRD pattern of the ZONF after calcinations at 700 C for 5 h. (e) TEM image of an
individual ZONF. (f) Bright field HRTEM image of the NF; inset is the corresponding EDS of the NF. (g) Corresponding SAED of the NF.
MW 1 300 000) were purchased from Sigma-Aldrich. Glucose, cholesterol, L-cysteine (L-Cys), ascorbic acid (AA), urea,
and citric acid were purchased from Sinopharm Chemical
Regent Co., Ltd. Other chemicals were of analytical-regent grade
without further purification. The pH of 0.1 M phosphate buffer
(PB) solution was adjusted by HNO3 and NaOH. Glucose stock
solution was kept for at least 24 h after preparation for
mutarotation. All solutions in the testing were prepared using
deionized water. The electrochemical experiments were performed at room temperature utilizing an electrochemical workstation (CHI660C) with a three-electrode mode: the modified
gold electrode was used as the working electrode, with Hg/
Hg2SO4 as the reference electrode, and silver as the counter
electrode. The pH of the solution was measured in real time by
a pH meter. The as-synthesized NFs were characterized by a
scanning electron microscope (SEM-6301F), X-ray diffraction
(XRD), a high-resolution transmission electron microscope
HRTEM (JEM-2011), and a Keithley 2400 sourcemeter.
II.2. Synthesis of NFs through Electrospinning. Electrospinning provided a simple and versatile method for producing
polymer fibers. A precursor polymer solution was prepared
containing 0.5 g of 10% zinc acetate (99.0%) and 0.26 g of
PVP in 0.7 g of ethanol. In a typical procedure, the precursor
polymer solution was loaded into a plastic syringe equipped
with a stainless-steel needle, and the distance between the needle
tip and the collector was 10 cm. The needle was connected to
a high-voltage power supply (operated at 15 kV) during
electrospinning, after which the zinc acetate/PVP solutions were
electrospun. The solution ejected from the tip of the needle
travels through the air to its target medium and accumulates as
a nonwoven fiber mat. The collector used here was composed
of two conductive substrates separated by a void gap as shown
in Figure 1a. Then the as-synthesized NFs were transferred onto
the as-prepared cleaned silicon substrate. By high-temperature
calcinations of the above precursor fibers, ZONFs were successfully obtained.
II.3. Fabrication of Single-NF-Based Glucose Biosensor.
To fabricate the biosensor, the as-prepared individual ZONF
after calcinations at 700 C is transferred to a conventional gold
electrode (with 3 mm diameter) under a high-resolution
microscope. The as-prepared ZONF/gold electrode is then
wetted by PB solution and dried in air for 2 h. A 5 L portion
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Ahmad et al.
Figure 3. (a) Schematic diagram of the modified gold electrode and the mechanism of the glucose sensing on the modified electrode. (b) Cyclic
voltammograms of the bare and modified gold electrode without and with 100 M glucose in pH 7.0 PB solution. (c) Cyclic voltammograms of
the biosensor in PB solution (pH 7.0) containing 100 M glucose at a scan rate of (a) 100 mV, (b) 80, (c) 50, and (d) 20 mV s-1.
Figure 4. (a) Amperometric response of the biosensor based on ZONF to different concentrations of glucose at 0.8 V in a stirring pH 7.0 PB
solution. Inset (bottom right): Response of biosensor with successive addition of glucose showed low LOD. (b) Calibrated curve of the biosensor
with successive addition of glucose. (c) Effect of interfering species to the response of the biosensor. (d) Amperometric response of the ZONFbased glucose biosensor in PB solution with increasing pH containing 0.1 mM glucose. (e) Long-term stability of the biosensor. (f) Temperature
profile of the biosensor in PB solution with 10 M glucose.
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Ahmad et al.
TABLE 1: Comparison of the Performance Parameters of Glucose Biosensor Based on Single ZONF with Other ZnO
Nanostructures
electrode materials
sensitivity/
A mM-1 cm-2
ZnO/MWNTs
ZnO nanocombs
ZnO nanorods
ZnO nanodisks
ZnO nanotubes
ZnO/PVP NF
50
15.33
23.1
16.9
30.85
70.2
linear range/mM
response time/s
ref.
<10
<5
<10
<6
<4
0.4
0.8
0.8
0.6
0.8
0.8
0.25
20
10
10
10
1.0
29
31
27
31
30
present work
0.1-16
0.01-3.45
0.01-4.2
0.25-19
JP102505G