Biosensors and Bioelectronics 25 (2010) 1402–1407

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Biosensors and Bioelectronics

journal homepage: www.elsevier.com/locate/bios

A highly sensitive nonenzymatic glucose sensor based on CuO

nanoparticles-modified carbon nanotube electrode
Liao-Chuan Jiang, Wei-De Zhang ∗
Nano Science Research Center, School of Chemistry and Chemical Engineering, South China University of Technology,
381 Wushan Road, Guangzhou 510640, People’s Republic of China

a r t i c l e i n f o a b s t r a c t

Article history: In this report, a novel type of cupric oxide (CuO) nanoparticles-modified multi-walled carbon nan-
Received 9 August 2009 otubes (MWCNTs) array electrode for sensitive nonenzymatic glucose detection has been fabricated.
Received in revised form 3 October 2009 The morphology of the nanocomposite was characterized by transmission electron microscopy and X-
Accepted 23 October 2009
ray diffraction. The electrochemical performance of the CuO/MWCNTs electrode for detection of glucose
Available online 30 October 2009
was investigated by cyclic voltammetry and chronoamperometry. The CuO/MWCNTs electrode showed
much higher electrocatalytic activity and lower overvoltage than the bare MWCNTs electrode towards
Keywords:
oxidation of glucose. At an applied potential of +0.40 V, the CuO/MWCNTs electrode presented a high
Cupric oxide
Carbon nanotubes
sensitivity of 2596 ␮A mM−1 cm−2 to glucose. In addition, linear range was obtained over a concentration
Electroanalysis up to 1.2 mM with a detection limit of 0.2 ␮M (signal/noise = 3). The response time is about 1 s with addi-
Glucose tion of 0.10 mM glucose. More importantly, the CuO/MWCNTs electrode is also highly resistant against
Sensor poisoning by chloride ion, and the interference from the oxidation of common interfering species such
as ascorbic acid, dopamine, uric acid and carbohydrate compounds is effectively avoided. In addition, the
CuO/MWCNTs electrode was also used to analyze glucose concentration in human serum samples. The
CuO/MWCNTs electrode exhibits an enhanced electrocatalytic property, low working potential, high sen-
sitivity, excellent selectivity, good stability, and fast amperometric sensing towards oxidation of glucose,
thus is promising for the future development of nonenzymatic glucose sensors.
© 2009 Elsevier B.V. All rights reserved.

1. Introduction have such drawbacks as low selectivity, high cost, or poisoning of

Since reliable and fast determination of glucose is important in
many areas such as biotechnology, clinical diagnostics and food
industry, the development of electrochemical glucose sensor has
attracted extensive attention (Wang, 2008; Wang, 2005; Rivas et al.,
2007; Lin et al., 2004). Glucose oxidase (GOx) has been widely used
to construct various amperometric biosensors for glucose detec-
tion, due to its high sensitivity and selectivity to glucose (Lim et al.,
2005; Tang et al., 2004; Hocevar et al., 2004; Umar et al., 2009;
Zhang et al., 2008). However, there are some disadvantages of
the enzyme-modified electrodes, such as instability, high cost of
enzymes, complicated immobilization procedure, critical operat-
ing situation, etc. Therefore, considerable attention has been paid
to develop nonenzymatic electrodes to solve these problems. Noble
metals (Li and Zhang, 2008; Park et al., 2003; Li et al., 2007), metal
alloys (Wang et al., 2008; Sun et al., 2001), and metal nanopar-
ticles (Rong et al., 2007; Kang et al., 2007; Cui et al., 2006; Jena
and Raj, 2006) have been extensively investigated in the develop-
ment of nonenzymatic glucose sensors. However, these electrodes
∗ Corresponding author. Tel.: +86 20 8711 4099; fax: +86 20 8711 2053.
E-mail address: zhangwd@scut.edu.cn (W.-D. Zhang).
chloride ion, which greatly limit their applications. Therefore, the
development of a cheap, highly selective, fast and reliable nonen-
zymatic glucose sensor is still imperatively demanded (Wang et al.,
2007; Chen et al., 2008; Jiang and Zhang, 2009).
Recently, carbon nanotubes (CNTs) have been rapidly becoming
electrode material due to their high surface area, unique structures,
excellent electrical conductivity, ultra-strong mechanical proper-
ties and high stability (Gooding, 2005; Agui et al., 2008). On the
other hand, as a p-type semiconductor with a narrow band gap
of 1.2 eV, CuO has been widely studied because of its numerous
applications in catalysis, semiconductors, gas sensors, biosensors,
and field transistors (Chen et al., 2003; Chowdhuri et al., 2004;
Luque et al., 2005; Zeng et al., 2008; Zheng et al., 2000). Some
efforts have been made on amperometric determination of glucose
using nanostructured CuO. For instance, CuO nanowires on a Cu rod
were used as electrode for detection of glucose with high sensitivity
(Zhuang et al., 2008). However, the synthesis of CuO nanowires is
tedious and time-consuming. In addition, exposing the active cop-
per substrate to the environment will affect the performance of the
sensor. In another work, CuO/Cu(OH)2 nanoparticles deposited on
graphite-like carbon films showed enhanced sensitivity and sta-
bility for glucose sensing (You et al., 2002). Most recently, the
existence of CuO as impurity in carbon nanotubes was claimed to be

0956-5663/$ – see front matter © 2009 Elsevier B.V. All rights reserved.
doi:10.1016/j.bios.2009.10.038
 L.-C. Jiang, W.-D. Zhang / Biosensors and Bioelectronics 25 (2010) 1402–1407
responsible for electrocatalytic activity of glucose oxidation while
using carbon nanotubes as electrode materials (Auley et al., 2008).
In the present work, vertically aligned multi-walled carbon nan-
otubes (MWCNTs) arrays on Ta foils were used as substrate for
deposition of CuO nanoparticles by magnetron sputtering depo-
sition. Well-aligned MWCNTs possess certain advantages because
they can provide more landing sites and have space between each
tube that would be favorable for dispersing nanoparticles com-
pared to disordered CNTs powder. Modification of MWCNTs with
CuO nanoparticles greatly improved the electrocatalytic properties
of glucose oxidation and detection. Compared with the MWCNTs
electrodes, the CuO/MWCNTs electrode presents high sensitivity,
excellent selectivity, low potential, high stability, and fast ampero-
metric response to the detection of glucose, which is promising for
the development of nonenzymatic glucose sensors.
2. Experimental
2.1. Reagents and materials
d-(+)-Glucose, dopamine (DA), l-ascorbic acid (l-AA), uric
acid (UA), d-fructose, lactose and sucrose were purchased
from Alfa Aesar and were used as-received. Bis-acetaldehyde-
oxalydihydrazone was obtained from Damao Chemical Reagent
(Tianjin, China). Deionized water (>18.4 M cm−1 ) was used for all
solutions’ preparation. All other reagents were of analytical grade
and used without further purification. The electrochemical mea-
surements were performed in 0.10 M NaOH solution.
2.2. Apparatus
CuO modified MWCNTs were observed by transmission elec-
tron microscope (TEM). Electrochemical experiments were carried
out on a CHI 660C electrochemical workstation (Shanghai Chen-
hua, China) with a conventional three-electrode setup, with
CuO/MWCNTs as working electrode, an Ag/AgCl (3 M KCl) electrode
and a platinum wire as reference electrode and counter electrode,
respectively. A magnetic stirrer and a stirring bar provided con-
vective mass transport. The content of CuO in the CuO/MWCNTs
electrode was determined by photometric method using a UV–vis
spectrophotometer (Hitachi U-3010).
2.3. Preparation of the CuO/MWCNTs electrode
Vertically aligned MWCNTs arrays on Ta foils (about
3 mm × 3 mm in area) were prepared by catalytic chemical
vapor deposition (Zhang et al., 2005, 2002a,b). The as-prepared
MWCNTs on the small Ta foils were coated with CuO by sputtering
deposition with a copper target at a power of 250 W for 6.5 min
under the flow of 300 mL min−1 Ar and 100 mL min−1 O2 . In
this procedure, the sputtered Cu was oxidized to cupric oxide
by oxygen and deposited on the MWCNTs. The Ta plate with
MWCNTs (with or without CuO) was connected to the surface of
a Cu electrode using conductive silver paint (Structure Probe Inc.,
USA). The edge of the Ta plate and Cu electrode was insulated by
pasting with nail enamel (Maybelline, NY, USA). These MWCNTs
on the Cu electrode were used as the working electrode and for
further characterization.
3. Results and discussion
3.1. Characterization of the CuO/MWCNTs nanocomposite
The morphology of the prepared vertically aligned MWCNTs
and CuO/MWCNTs is depicted in Fig. 1A and B. One can see
1403
that the MWCNTs are obviously thicker after being coated. TEM
observations give more detailed structural characteristics of the
CuO/MWCNTs nanocomposite. From Fig. 1C, it can be seen that the
CuO nanoparticles are coated homogeneously on the walls of the
carbon nanotubes. The right up inset in Fig. 1C is the selected area
electron diffraction pattern of the nanoparticles, indicating multi-
crystal characteristic of the CuO. Loading of CuO nanoparticles on
MWCNTs arrays will provide large surface area for electrocatalytic
reaction (Reitz et al., 2008). The corresponding high-resolution TEM
(HRTEM) image in Fig. 1D shows a clear interplanar spacing of
0.131, 0.141 and 0.151 nm, which corresponds to the (3 1 1), (0 2 2)
and (2 0 2) planes of the CuO (JCPDS 45-0937). It confirms that the
prepared CuO/MWCNTs nanocomposite is well crystallized with
a polycrystal structure. The crystal structure of the CuO/MWCNTs
nanocomposite was further characterized by X-ray diffraction anal-
ysis, as shown in Fig. 1E. The XRD pattern shows reflections of
tantalum substrate and CuO. The peaks with 2 of 38.47◦ , 55.55◦ ,
69.58◦ and 94.94◦ are assigned to the tantalum substrate (JCPDS
04-0788), and the peaks with 2 of 35.50◦ and 38.73◦ correspond
to monoclinic CuO (JCPDS 45-0937). No reflections for Cu metal or
metal carbides are observed, indicating the Cu was totally oxidized
to CuO during the sputtering deposition. The content of CuO in the
CuO/MWCNTs was determined by photometric method using bis-
acetaldehyde-oxalydihydrazone as a chromogenic agent. The CuO
content in a CuO/MWCNTs electrode with area of 3 mm× 3 mm was
18 ␮g.
3.2. Electrocatalysis of glucose at the CuO/MWCNTs electrode
The electrocatalytic activity of the MWCNTs and CuO/MWCNTs
electrodes towards the oxidation of glucose in an alkaline medium
was demonstrated by CVs. Fig. 2 displays the CVs of 1.0 mM glucose
in 0.10 M NaOH at the bare MWCNTs and CuO/MWCNTs electrodes,
respectively. Fig. 2 (curve a) shows that the MWCNTs electrode
exhibits a capacitive charging response with a large oxidative
tail corresponding to the onset of water breakdown observed at
+0.65 V. Fig. 2 (curve b) shows that only a small current for glucose
oxidation and no peak is observed at the bare MWCNTs electrode
(Ye et al., 2004). From Fig. 2 (curve d), one can see that a broad
reduction peak with a peak potential of about +0.60 V is observed
at the CuO/MWCNTs electrode in the alkaline blank solution. The
reductive peak might correspond to a Cu(II)/Cu(III) redox couple
(Xu et al., 2006; Farrell and Breslin, 2004). However, the corre-
sponding oxidation peak of Cu(II)/Cu(III) within +0.40 to +0.80 V
is not clearly observed at the CuO/MWCNTs electrode, which could
be overlaid by the oxidation peak of water-splitting. This result is
similar to previous reports (Kang et al., 2007; You et al., 2002).
Upon addition of 1.0 mM glucose, a single oxidative peak, cor-
responding to the irreversible oxidation of glucose, is observed
at the CuO/MWCNTs (Fig. 2, curve c) electrode. Compared with
the MWCNTs electrode, the CuO/MWCNTs electrode displays a
substantially negative shift in peak potential with oxidation start-
ing at about +0.25 V and peak potential at +0.38 V. The obvious
decrease in the anodic overpotential suggests a high electrocat-
alytic activity of the CuO/MWCNTs nanocomposite in the direct
oxidation of glucose. The overpotential shifted negatively, which
can be attributed to a kinetic effect by an increase in the electroac-
tive surface area and the rate of electron transfer from glucose to
the CuO/MWCNTs electrode (Zhuang et al., 2008). In addition, it
also can be seen that the peak current drastically increases and con-
tinues to increase at a potential of about +0.60 V. This may result
from the proposed involvement of Cu(II)/Cu(III) surface species in
the oxidation of glucose (Kang et al., 2007; Farrell and Breslin,
2004).
Furthermore, the CVs of glucose solution at different scan rates
were recorded, as shown in Fig. 3. The anodic peak current (Ipa ) of
1404 L.-C. Jiang, W.-D. Zhang / Biosensors and Bioelectronics 25 (2010) 1402–1407

Fig. 1. SEM images of the bare (A) and CuO-coated MWCNTs with part of the nanotubes being peeled off (B), and TEM (C) and HRTEM (D) images of carbon nanotube coated
by CuO; X-ray diffraction pattern of the CuO/MWCNTs nanocomposite (E). Left bottom inset in (C) is a TEM image of bare carbon nanotube, right up inset in (C) is the selected
area electron diffraction pattern of CuO.

oxidation of glucose is proportional to the scan rate. A good lin-
earity between scan rate and peak current was obtained within
the range of 20–200 mV s−1 , as indicated in the inset of Fig. 3. The
linear regression equation, Ipa (␮A) = 65.4242 + 1.2867 V (mV s−1 ),
with the correlation coefficient (R) of 0.9944, was obtained. The
result indicates that the electrochemical kinetics is controlled by
the adsorption of glucose (Chen et al., 2008).
3.3. Amperometric response of the CuO/MWCNTs electrode to
glucose
Fig. 4A shows typical amperometric response curves of glucose
in alkaline solution at the MWCNTs and CuO/MWCNTs elec-
trodes. At +0.40 V, the MWCNTs electrode exhibits low response
with the addition of 1.0 mM glucose into 0.10 M NaOH solution.
 L.-C. Jiang, W.-D. Zhang / Biosensors and Bioelectronics 25 (2010) 1402–1407 1405

Table 1
Comparison of the present CuO/MWCNTs electrode with other nonenzymatic glucose sensors.

Electrode Detection potential Sensitivity Linear range Detection limit Reference

−1 −2
Freestanding SWCNT film +0.6 V 248.6 ␮A mM cm 0.01–2.16 mM 0.01 mM Wang et al. (2007)
MnO2 /MWCNTs +0.30 V 33.19 ␮A mM−1 10 ␮M to 28 mM – Chen et al. (2008)
Mesoporous Pt +0.4 V 9.6 ␮A mM−1 cm−2 0–10 mM – Park et al. (2003)
Porous Au +0.35 V 11.8 ␮A mM−1 cm−2 2–10 mM 5 ␮M Li et al. (2007)
MWCNTs +0.20 V 4.36 ␮A mM−1 cm−2 2 ␮M to 11 mM 1 ␮M Ye et al. (2004)
Pt nanotubules +0.4 V 0.1 ␮A mM−1 cm−2 2–14 mM 1.0 ␮M Yuan et al. (2005)
Pt–Pb/CNTs +0.30 V 17.8 ␮A mM−1 cm−2 Up to 11 mM 1.8 ␮M Cui et al. (2007)
Au nanoparticles – 160 ␮A mM−1 cm−2 0.5–8 mM – Kurniawan et al. (2006)
CuO nanowires +0.33 V 0.49 ␮A mM−1 cm−2 0.4 ␮M to 2.0 mM 0.049 ␮M Zhuang et al. (2008)
Cu/MWCNTs +0.65 V 251.4 ␮A mM−1 cm−2 0.7–3.5 mM 0.21 ␮M Kang et al. (2007)
Cu nanoparticles +0.65 V – 1 ␮M to 5 mM 0.5 ␮M Xu et al. (2006)
Ni nanowires +0.55 V 1043 ␮A mM−1 cm−2 0.5 ␮M to 7 mM 0.1 ␮M Lu et al. (2009)
CuO/MWCNTs +0.40 V 2596 ␮A mM−1 cm−2 0.4 ␮M to 1.2 mM 0.2 ␮M This work

glucose sensor is shown in Fig. 4B. The regression equation, Ipa

(␮A) = −10.1688 + 233.5568c (mM), with R = 0.9983, was obtained.
The sensor displays a linear range of 0.4 ␮M to 1.2 mM glucose with
a sensitivity of 2596 ␮A mM−1 cm−2 , and a detection limit of 0.2 ␮M
(signal/noise = 3). The sensitivity of the CuO/MWCNTs electrode
is obviously higher than that of those using SWCNTs, MWCNTs,
and other nonenzymatic glucose sensors such as noble metals and
nanoparticles as electrodes, which is summarized in Table 1. This
can be attributed to the CuO nanoparticles loading on MWCNTs
arrays, which significantly increase the electrocatalytic active areas
and promote electron transfer in the oxidation of glucose (Reitz
et al., 2008). These experimental results show that the electro-
chemical sensor based on CuO/MWCNTs nanocomposite exhibits
the characteristics of high sensitivity, low detection limit and fast
response time.

Fig. 2. CVs of MWCNTs (a and b) and CuO/MWCNTs (c and d) electrodes in the

absence (a and d) and presence (b and c) of 1.0 mM glucose in 0.10 M NaOH. Scan
rate: 100 mV s−1 . Inset is the plot of oxidative peak in the range from +0.3 to +0.7 V
at the MWCNTs electrode.

The regression equation is Ipa (␮A) = 0.1163 + 1.7571c (mM), with

R = 0.9994. In contrast, the CuO/MWCNTs electrode displays much
higher current response than the MWCNTs electrode, even the
concentration of glucose at the MWCNTs electrode is 10 times
that of the CuO/MWCNTs electrode. A well-defined, stable and
fast amperometric response could be observed at +0.40 V with
successive addition of 0.10 mM glucose into 0.10 M NaOH at an
interval of 50 s. It is noted that the noise increased with concen-
tration. This is because more and more intermediate species are
adsorbed onto the electrode with increase of glucose’s concentra-
tion and prolonging of reaction time. The calibration curve for the

Fig. 3. (a–e) CuO/MWCNTs electrode at scan rates of 20–200 mV s−1 in 0.10 M NaOH
solution containing 1.0 mM glucose. Inset is the plot of oxidative peak current with
scan rate.
Fig. 4. (A) Amperometric response of (a) MWCNTs and (b) CuO/MWCNTs electrodes
at +0.40 V upon addition of glucose in a step of 1.0 and 0.10 mM, respectively for each
current step. (B) The calibration curve of current vs. concentration of glucose at the
CuO/MWCNTs electrode. The error bars denote the standard deviation of triplicate
determination of each concentration of glucose.
1406 L.-C. Jiang, W.-D. Zhang / Biosensors and Bioelectronics 25 (2010) 1402–1407
Fig. 5. (A) Long-term stability of the CuO/MWCNTs sensor at room temperature.
(B) Flow injection amperometric response to injections of 1.0 mM glucose, 0.10 mM
interferents of DA, AA, UA, fructose, lactose and sucrose at the CuO/MWCNTs elec-
trode under +0.40 V. The inset is the plot of current response of interferents. The error
bars denote the standard deviation of triplicate determination of each interferent.
3.4. Reproducibility, stability and anti-interference property of
the CuO/MWCNTs electrode
The reproducibility and stability of the sensor were evalu-
ated. Five CuO/MWCNTs electrodes were investigated at +0.40 V to
compare their amperometric current responses. The relative stan-
dard deviation (R.S.D.) was 2.8%, confirming that the preparation
method was highly reproducible. Nine successive measurements
of glucose on one CuO/MWCNTs electrode yielded an R.S.D. of
3.4%, indicating that the sensor was stable. The long-term sta-
bility of the sensor was also evaluated by measuring its current
response to glucose within a 30-day period (Fig. 5A). The sen-
sor was exposed to air and its sensitivity was tested every 2
days. The current response of the CuO/MWCNTs electrode was
approximately 90% of its original counterpart, which can be mainly
attributed to the chemical stability of CuO in basic solution. Fur-
thermore, most nonenzymatic glucose sensors based on metallic
Cu nanoparticles lose their activity easily due to poisoning by chlo-
ride ion, which is because of the reaction between Cu and Cl−
and formation of complexation. This will reduce the electrocat-
alytic activity of the Cu electrode (Yuan et al., 2009). In contrast,
CuO is difficult to form complexation with Cl− since the elec-
tronegativity of O is higher than that of Cl. The current response
of the CuO/MWCNTs electrode was also examined by adding 1.0 M
KCl in the solution as supporting electrolyte. The experimental
result shows that the peak current of the CuO/MWCNTs electrode
towards oxidation of glucose remains almost unchanged (figure not
shown), indicating that the electrode is not poisoned by chloride
ion.
Table 2
Amperometric determination of glucose in human blood serum samples.
Sample Concentration (mM) R.S.D. (%)a Added (mM) Recovery (%)
1 12.2 1.7 0.1 97
2 4.9 4.3 0.1 98
3 4.7 3.8 0.1 96
4 5.3 1.3 0.1 98
5 4.8 1.9 0.1 95
a
R.S.D. (%) calculated from three separate experiments.
Anti-interference property is also an important factor for sen-
sors. Because the easily oxidative species such as dopamine,
ascorbic acid, uric acid and other carbohydrate compounds usu-
ally co-exist with glucose in human blood, the electrochemical
response of the interfering species was also examined at the
CuO/MWCNTs electrode, as shown in Fig. 5B. Considering the con-
centration of glucose is at least 30 times of interfering species in
human blood, the interference experiment was carried out by suc-
cessive injection of 1.0 mM glucose and 0.10 mM interfering species
in 0.10 M NaOH solution. A well-defined glucose response was
obtained, while insignificant responses were observed for interfer-
ing species. Compared to glucose, the interfering species yielded
current response ranging from 1.5% (uric acid) to 2.7% (dopamine)
at +0.40 V. It can be concluded that small amount of DA, AA, UA
and other carbohydrate compounds, such as fructose, lactose and
sucrose can be neglected. In other words, the CuO/MWCNTs elec-
trode shows high selectivity for glucose detection.
3.5. Human serum samples measurement
In an attempt to explore the CuO/MWCNTs electrode for prac-
tical applications, the sensor was applied to determine glucose in
human blood serum samples of diabetic and healthy people. 40.0 ␮L
of serum sample was added to 10.0 mL 0.10 M NaOH solution, and
the current response was recorded at +0.40 V. Table 1 displays the
determination result of five samples including one diabetic and four
healthy people. The glucose concentration in the serum of the dia-
betic is as high as 12.2 mM, while it is only 4.7–5.3 for the healthy
ones. The recovery of glucose was determined by standard addi-
tion of pure glucose to the solutions containing the serum samples
and the corresponding results are given in Table 2. One can see
that the CuO/MWCNTs sensor also gives exact recovery (≥95%).
The results demonstrated here reveal the potential applications of
the CuO/MWCNTs sensor for determination of glucose in biological
fluids.
4. Conclusions
We have successfully deposited CuO nanoparticles on the ver-
tically aligned MWCNTs arrays by magnetron sputtering. The
CuO/MWCNTs nanocomposite electrode is used to construct a
novel nonenzymatic glucose sensor, which presents many attrac-
tive analytical features such as superb electrocatalytic activity,
high sensitivity, strong stability, good reproducibility, and excel-
lent selectivity as well as quick response. This is because of
the improvement of electroactive surface area and the syner-
gistic electrocatalytic activity resulting from the combination of
CuO nanoparticles and MWCNTs. The CuO/MWCNTs electrode
can also be used as an amperometric sensor for routine analysis
of glucose in human serum samples. These experimental results
indicate that the CuO/MWCNTs nanocomposite electrode holds
the prospect for effective nonenzymatic determination of glucose
at low concentrations and detection limit, but with very high
sensitivity.
 L.-C. Jiang, W.-D. Zhang / Biosensors and Bioelectronics 25 (2010) 1402–1407
Acknowledgements
The financial support by National Natural Science Foundation of
China (no. 20773041), the Research Fund for the Doctoral Program
of Higher Education (no. 20070561008), and the high technology
research program, Ministry of Science and Technology (MOST) of
China (2008AA06Z311) to the work was gratefully acknowledged.
The authors would like to extend sincere thanks to Nanfang Hospi-
tal in Guangzhou for donating the blood serum samples.
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