Sensors and Actuators B 104 (2005) 68–74

Amperometric glucose biosensor based on immobilization

of glucose oxidase in electropolymerized o-aminophenol
film at copper-modified gold electrode
Dawei Pan, Jinhua Chen∗ , Shouzhuo Yao, Lihua Nie, Jianjun Xia, Wenyan Tao
State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering,
Hunan University, Changsha 410082, PR China
Received 3 January 2004; received in revised form 27 April 2004; accepted 27 April 2004
Available online 7 June 2004

Abstract

Amperometric glucose biosensor is developed, based on immobilization of glucose oxidase (GOD), in an electrochemically polymerized
non-conducting poly(o-aminophenol) (POAP) film at copper (Cu)-modified gold (Au) electrode. The rough surface and the ability to
electrochemically oxidize glucose of Cu nanoparticles result in the improvement of the detection limit and the increase of the maximum
response current and sensitivity. The biosensor based on Au/Cu/POAP/GOD electrode has two times lower detection limit, three times
larger maximum current and 2.5 times higher sensitivity than those of the biosensor based on Au/POAP/GOD electrode. Additionally, the
fast response time, large response current and good selectivity for ascorbic acid, uric acid and acetaminophen can also be obtained. On the
other hand, effects of electrochemical deposition time of Cu, applied potential used in the determination and electroactive compounds on
the amperometric response of the enzymatic sensor were investigated and discussed. Excellent reproducibility and stability of biosensor
are also observed.
© 2004 Elsevier B.V. All rights reserved.

Keywords: Glucose oxidase; Poly(o-aminophenol); Copper; Electrochemical polymerization; Biosensor

1. Introduction enzyme-catalysed oxidation of glucose by dissolved oxy-

The development of novel biosensor for the fast, sensitive
and selective quantification of analytes of biomedical inter-
est has received considerable attention, and electrochemical
biosensors have been successfully used for quantifying
these analytes [1]. Glucose is a key metabolite for living
organisms, especially in the case of patients suffering di-
abetes. In general, glucose oxidase (GOD) is selected as a
model enzyme, since it is well-studied, inexpensive, stable
and practically applied [2–5]. The final goal of glucose elec-
trochemical biosensor lies in designing high-performance
sensors with appropriate characteristics such as sensitivity,
selectivity, response time, linearity, stability and repro-
ducibility [6].
Most of the electrochemical amperometric biosensors
for glucose are based on the electrochemical oxidation of
hydrogen peroxide, which is formed in the course of the
∗ Corresponding author. Tel.: +86 731 8821818; fax: +86 731 8821818.
E-mail address: chenjinhua@hnu.cn (J. Chen).
gen [7]. From the viewpoint of chemical sensors, hydrogen
peroxide can be detected by oxidation at anodic potentials
(>+0.6 V versus Ag/AgCl) [8,9]. But at this relatively high
potential, there may be interferences from other oxidasable
species such as ascorbic acid (AA), uric acid (UA) and
acetaminophen (AP). It results in difficulty for the quanti-
tative analysis of the produced hydrogen peroxide and the
enzyme substrate.
One approach to avoid interferents developed in recent
decade years is related to the use of selective membranes
that are permeable for hydrogen peroxide but non-permeable
for interference species [10]. Many of the known selective
membranes, such as cellulose acetate membrane, do not en-
sure a satisfactory avoidance of interferents. Therefore, seri-
ous research activities should be taken in creating improved
selective membranes for selectivity biosensors. In general,
the selectivity of the polymer depends on its pore size and
charges [11,12]. The non-conducting polymers, due to their
good selectivity and fast response, have attracted much inter-
est in the development of the biosensors. The glucose biosen-
sors based on the non-conducting poly(o-phenylenediamine)

0925-4005/$ – see front matter © 2004 Elsevier B.V. All rights reserved.
doi:10.1016/j.snb.2004.04.100
 D. Pan et al. / Sensors and Actuators B 104 (2005) 68–74
[13], poly(ethacridine) [14], overoxidized polyprrole [15],
poly(phenol) [16] and poly(o-aminophenol) (POAP) [17]
films have been reported and showed the good selectivity.
Although non-conducting polymer films exhibit excellent
selectivity properties, their use is still a matter of concern
because of their relative high detection limit and the low re-
sponse current. To increase the response current, ferrocene,
a satisfactory electron transfer mediator for the amperomet-
ric hydrogen peroxide sensor, has been used in the biosensor
based on non-conducting polymer films of phenol and its
derivatives and an increasing response current of hydrogen
peroxide can be observed [18,19].
In this paper, an amperometric glucose biosensor based
on GOD immobilized in electrochemical polymerized POAP
film at Cu-modified gold (Au) electrode has been developed.
o-Aminophenol (o-AP) can be electropolymerized to form
electroinactive film when the pH of the solution is over 3
[20]. POAP film, a non-conducting film which acted as an
effective barrier to protect the electrode from fouling, was
successfully used in the development of hydrogen peroxide,
uric and glucose biosensor [21–23]. Additionally, copper is
selected in this paper in order to increase the response current
because it can electrochemically oxidize glucose [24] and is
relatively inexpensive, commercially available in a number
of forms and the method for its deposition is well established
[25,26]. The experimental conditions related to the prepara-
tion and characterizations of the sensor have also been stud-
ied in detail. The sensor exhibits excellent performances,
such as relative low detection limit, short response time,
large current density and high sensitivity. Additionally, the
effect of the interferents (AA, UA and AP) can be decreased
significantly due to selective permeability of POAP film.
2. Experimental
2.1. Reagents and apparatus
Glucose oxidase (EC 1.1.3.4, Type II from Aspergillus
niger, 50 000 U/g) was purchased from Amresco (USA).
Glucose was obtained from ICN Biomedicals, Inc. (USA).
Glucose stock solutions were allowed to mutarotate at room
temperature overnight before use. Hydrogen peroxide, o-AP
(99%), copper chloride, AA, UA and AP were used as re-
ceived (Chemical Reagent Company of Shanghai, China).
0.2 M acetate solution and 1/15 M phosphate solution were
used as buffer solutions. All other chemicals were analytical
grade. Double-distilled water was used throughout.
All electrochemical experiments were carried out in a con-
ventional three-electrode cell controlled by CHI 660 Elec-
trochemical Work Station (Chenghua Instrument Company
of Shanghai, China). A gold wire (diameter 0.2 mm) encap-
sulated in epoxy resin was used as working electrode. A
platinum foil was applied as the counter electrode and a satu-
rated calomel electrode (SCE) served as reference electrode.
All potential values given below refer to SCE. Amperomet-
69
ric measurements were carried out under stirred condition
and the response current was marked with the change value
between the steady-state current and background current.
All experiments were performed at room temperature. The
micrographs and element composition of Au/Cu electrodes
were investigated by scanning electron microscopy (SEM,
JSM 5600 LV, operating at 20 kV) and energy-dispersive
X-ray spectroscopy (EDS, Vantage 4105, NORAN), respec-
tively.
2.2. Electrochemical deposition of Cu and preparation of
Au/Cu/POAP/GOD electrode
Prior to electrochemical deposition of Cu, the Au work-
ing electrode was cut to get bare fresh surface, rinsed with
double-distilled water and ethanol, and then followed by
cleaning in an ultrasonic bath. The electrode was then ac-
tivated by continuously scanning the potential between 0.0
and 1.5 V at 0.05 V s−1 in 1 M H2 SO4 solution and washed
by double-distilled water. Cu was electrochemically de-
posited by potentiostatic method in a 0.2 M acetate buffer
solution (pH 4.0) containing 5.0 mM CuCl2 .
The Cu-modified Au electrode was placed in 0.2 M deaer-
ated acetate buffer solution (pH 5.0) containing 200 U/ml
GOD + 5.0 mM o-AP monomer. The electrochemical poly-
merization of POAP/GOD film was carried out by cycling
in the potential range from 0.0 to 0.8 V at a scan rate of
50 mV s−1 . For comparison, Au/POAP/GOD electrode was
also fabricated at bare Au electrode in the same way. All
films used in the following experiments were grown for 15
cycles except as noted. The resulting enzyme electrodes were
thoroughly washed by double-distilled water and stored in
phosphate buffer solution (pH 7.0) below 4 ◦ C for future use.
3. Results and discussion
3.1. Preparation and properties of Cu-modified Au
electrode
The Cu-modified Au electrode is fabricated by applying
0.0 V for 180 s in a 0.2 M acetate buffer solution (pH 4.0)
containing 5.0 mM CuCl2 . The micrographs and element
composition of Au/Cu electrode have been investigated by
SEM and EDS, and the corresponding results are shown in
Fig. 1. From Fig. 1(a), the diameter of Cu particle is about
200 nm. Cu particles with spherical shape lie on the electrode
surface. From Fig. 1(b), EDS result indicates that Au and
Cu are the major elements, which implies that the particles
on the Au electrode are Cu.
Cyclic voltammograms of Cu-modified Au electrode in
1/15 M phosphate buffer solution (pH 7.0) with and without
3 mM hydrogen peroxide are shown in Fig. 2. From Fig. 2,
a pair of Cu redox peaks at 0.28 and 0.12 V can be observed
for the case of the absence of hydrogen peroxide. But for
the case of the addition of hydrogen peroxide, the disap-
70 D. Pan et al. / Sensors and Actuators B 104 (2005) 68–74
Fig. 1. The SEM image (a) and EDS pattern (b) of the Au/Cu electrode.
Magnification 20 000×.
pearance of the copper oxidation peak can be observed. This
implies that the chemical oxidation of copper by hydrogen
peroxide occurs. This phenomenon is similar to that reported
in literature [27]. Additionally, it can be observed from
Fig. 2 that the oxidation of hydrogen peroxide is detected
at Cu-modified Au electrode at the potential of +0.4 V or
higher.
Fig. 2. Cyclic voltammograms of the Au/Cu electrode in a 1/15 M phos-
phate buffer solution (pH 7.0) with (solid line) and without (dot line)
3 mM hydrogen peroxide. The potential range is from −0.4 to 0.8 V and
the scan rate is 50 mV s−1 .
Fig. 3. Cyclic voltammograms for the electropolymerization of
POAP/GOD film in a deaerated and unstirred 5.0 mM o-AP + 200 U/ml
GOD + 0.2 mM acetate buffer solutions (pH 5.0) in the potential range
of 0.0–0.8 V at 50 mV s−1 . Numerals indicate the number of cycles.
3.2. Electrochemical co-polymerization of o-AP and GOD
on Cu-modified Au electrode
Fig. 3 presents the cyclic voltammograms for the electro-
chemical co-polymerization of 5 mM o-AP and 200 U/ml of
GOD in acetate buffer solution (pH 5.0) at the Cu-modified
Au electrode. Compared with the electrochemical polymer-
ization of o-AP [17], a reduction peak of copper appears
obviously at near 0.2 V at the first cycle. Furthermore,
with the increase of the polymerization cycles, all the peak
currents decrease. These results imply that non-conducting
POAP/GOD film has been formed on the surface of the
Cu-modified Au electrode.
3.3. Influential factors on the response characteristic of
the Au/Cu/POAP/GOD electrode
The amperometric response characteristics of the enzyme
electrode are affected by the deposition time of copper, ap-
plied potential used in the determination and the electroac-
tive interferents. In this paper, the effects of these factors on
the behavior of the GOD electrode have been investigated
in detail and discussed as follows.
3.3.1. Effect of deposition time of copper
In general, the response of the biosensor should be af-
fected by the loading mass of Cu nanoparticle. The effect
of the different deposition time of copper on the response
current was investigated and the corresponding results are
shown in Fig. 4. From Fig. 4, it can be observed that the re-
sponse current is affected obviously by the loading mass of
Cu. The response current increases with the increase of the
Cu loading and the maximum value can be obtained at the
deposition time of 180 s. This may result from the loading
mass and particle size of Cu nanoparticles. When the depo-
sition time increases, the loading mass of Cu increases and
the response current of the biosensor increases due to the
 D. Pan et al. / Sensors and Actuators B 104 (2005) 68–74
Fig. 4. Effect of the deposition time of copper on the steady-state response
current of Au/Cu/POAP/GOD electrode in 1/15 M phosphate buffer solu-
tion (pH 7.0) containing 1 mM glucose (n = 3).
catalytic property of Cu nanoparticle. However, the particle
size of Cu will increase with the increase of the deposition
time and the corresponding catalytic activity of Cu particle
becomes low [28,29]. In the following experiments, 180 s
was selected as the Cu deposition time.
3.3.2. Effect of the applied potential
Fig. 5 shows the response current of the enzyme elec-
trode in 1/15 M phosphate buffer solution (pH 7.0) contain-
ing 1 mM glucose as a function of the applied potential,
which ranges from 0.5 to 0.9 V with 0.05 V increments.
From Fig. 5, when the applied potential is below 0.7 V,
the response current increases rapidly with the increase
of the applied potential. This means that the response of
the enzyme electrode in this potential range (<0.7 V) was
controlled by the electrochemical oxidation of hydrogen
peroxide [30,31]. When the potential is over 0.7 V, the re-
sponse current remains almost same, which is explained by
the rate-limiting process of enzyme kinetics and substrate
diffusion [30,31]. Considering that the higher applied po-
Fig. 5. Effect of applied potential on the response current of Au/Cu/POAP/
GOD electrode. Steady-state currents are measured in 1/15 M phosphate
buffer solution (pH 7.0) containing 1 mM glucose.
71
Fig. 6. Anodic response to the addition (indicated by the arrows) of
3 mM ascorbic acid at bare Au (a), Au/Cu (b) and Au/Cu/POAP/GOD (c)
electrodes in 1/15 M phosphate buffer solution (pH 7.0). Applied potential
is 0.7 V.
tential is, the easier the electroactive interferents are to be
oxidized and cause the extra response current and the easier
the polymer film is to be damaged, we set the potential of
0.7 V for the oxidation operation of the enzyme electrode.
3.3.3. Effect of the interferents
Fig. 6 presents the response current to ascorbic acid ob-
tained at various electrodes at 0.7 V. From Fig. 6, the large
response current of ascorbic acid can be observed obviously
at unmodified Au electrode and Cu-modified Au electrode,
whereas at the enzyme electrode modified by POAP film, the
response current decreases significantly. These results show
that the POAP-modified electrodes can avoid efficiently the
interference of the ascorbic acid. The similar situation can
also be observed for uric acid and acetaminophen. On the
other hand, in discussing the magnitude of the interference, it
is much more relevant to consider the magnitude of the inter-
ferents current relative to the analytical signal produced by
the analyte (glucose). The interference of electroactive com-
pounds to the glucose response was examined in the pres-
ence of their physiological normal level (0.1 mM for ascorbic
acid, 0.5 mM for uric acid and 0.1 mM for actaminophen, re-
spectively) [31] with glucose concentration of 5.6 mM. The
influence of ascorbic acid, uric acid and acetaminophen to
the glucose response was little under the testing conditions.
The ratio of IG+I to IG is 1.02 for ascorbic acid, 1.05 for uric
acid and 1.06 for acetaminophen, respectively. The sensor
based on Au/Cu/POAP/GOD electrode has practically good
selectivity. These results are due to the good selectivity of
POAP film [9,17,21].
72 D. Pan et al. / Sensors and Actuators B 104 (2005) 68–74
Fig. 7. The amperometric response of Au/Cu/POAP/GOD (a) and
Au/POAP/GOD (b) electrodes in a stirred phosphate buffer solution (pH
7.0) upon the injection of 1 mM glucose in each step (indicated by the
arrows). Applied potential is 0.7 V.
3.4. The comparison of the Au/Cu/POAP/GOD and
Au/POAP/GOD electrodes for the amperometric
determination of glucose
Fig. 7 illustrates that the typical current–time plots for the
Au/Cu/POAP/GOD and Au/POAP/GOD electrodes upon
the successive step addition of glucose were obtained by
injecting stock glucose solution. The response of these two
biosensors to the injection of glucose is very fast (within
5 s) due to the thin POAP film. This is also observed
for the other non-conducting polymer-modified electrode
[13,14]. Additionally, the response current of 1 mM glu-
cose on Au/Cu/POAP/GOD electrode is about 2.5 times
higher than that on the Au/POAP/GOD electrode. The
calibration curves of the response of the Au/POAP/GOD
and Au/Cu/POAP/GOD electrodes for different glucose
concentration at the applied potential of 0.7 V are pre-
sented in Fig. 8. From Fig. 8, the response current of the
biosensor based on Au/POAP/GOD electrode is linear with
glucose concentration up to 10 mM and then a plateau is
reached gradually at higher glucose concentration. The de-
tection limit of the biosensor is 0.02 mM. Compared with
Au/POAP/GOD biosensor, the Au/Cu/POAP/GOD biosen-
sor has lower detection limit (0.01 mM) and larger response
current. These may result from the following reasons: (a)
the surface coverage of enzyme on the Au/Cu/POAP/GOD
electrode is higher than that on the Au/POAP/GOD elec-
trode due to an increased surface by Cu particle, and (b)
the copper particles have the ability to oxidize the glucose.
However, the Au/Cu/POAP/GOD biosensor has narrower
linear range (up to 6 mM).
According to the Lineweaver–Burk form of the Michaelis–
Menten equation, the relation between the reciprocal of the
response current and the reciprocal of glucose concentration
app
is obtained. The apparent Michaelis–Menten constant KM
Fig. 8. Calibration curve of the response current of GOD biosensor to
glucose concentration in 1/15 M phosphate buffer solution (pH 7.0) at the
Au/Cu/POAP/GOD electrode (a) and the Au/POAP/GOD electrode (b).
Applied potential is 0.7 V.
and the maximum response current of the biosensor based
on the Au/POAP/GOD electrode are calculated and equal
to 22.8 mM and 0.15 mA cm−2 (0.047 ␮A), respectively.
The data of the maximum response current are similar to
that of GOD electrodes modified by other non-conducting
polymer films [14,16]. The sensitivity of biosensor obtained
from the slope of the initial linear part of the calibration is
4.7 mA M−1 cm−2 , which is similar to the reported value
[31]. Compared with Au/POAP/GOD electrode, the appar-
ent Michaelis–Menten constant of the Au/Cu/POAP/GOD
electrode is also calculated and equal to 17.8 mM. The
maximum current is 0.45 mA cm−2 (0.14 ␮A), which is
three times larger than that at the Au/POAP/GOD elec-
trode. The sensitivity of the Au/Cu/POAP/GOD electrode
is 12.6 mA M−1 cm−2 , which is almost 2.5 times larger
than that of Au/POAP/GOD electrode. These results imply
that the biosensor based on Au/Cu/POAP/GOD electrode is
actually valuable and sensitive.
3.5. Reproducibility, stability and sample analysis
The reproducibility of five Au/Cu/POAP/GOD electrodes
was estimated by the response to 1 mM glucose at the poten-
tial of 0.7 V. The results reveal that the sensor has satisfied
reproducibility with a mean change of the response current
of 4.1 nA and a relative standard deviation of 9.2%.
The stability of enzymatic biosensor in storage conditions
(phosphate buffer of pH 7.0 at 4 ◦ C) was investigated in the
same phosphate buffer solution containing 2.0 mM glucose.
The corresponding result shows that 72% response current
is still retained after 30 days. This result is similar to that of
POAP-modified GOD electrode [9,17] and implies that the
Au/Cu/POAP/GOD electrode is considerably stable. Good
stability may be attributed to the enzyme entrapped strongly
in the POAP film that is stable in the neutral medium.
Human plasma samples were assayed in order to demon-
strate the practical usage of the biosensor. Fresh plasma
 D. Pan et al. / Sensors and Actuators B 104 (2005) 68–74
Table 1
Glucose contents in human blood samples
Sample Determined by Measured by Bias (mM)
number hospital (mM) biosensor (mM)
1 3.64 3.82 +0.18
2 4.27 4.55 +0.28
3 5.80 6.12 +0.32
4 6.30 6.46 +0.16
5 9.07 8.71 −0.36
samples were first analyzed in the local hospital with
ASCA AG-II Chemistry System, Landmark, USA. The
samples were then re-assayed with the Au/Cu/POAP/GOD
electrode. Plasma sample (0.5 ml) was added into 2 ml of
phosphate buffer solution (pH 7.0) and the response was
obtained at 0.7 V. The contents of glucose in blood can then
be calculated from the calibration curve. The results, which
are listed in Table 1, are satisfactory and agree closely with
those measured by the biochemical analyzer in hospital.
4. Conclusions
Immobilization of GOD is introduced by the electro-
chemical co-polymerization of GOD and o-AP monomer
at the Cu-modified Au electrode in the weak acidic
medium. Compared with the Au/POAP/GOD sensor, the
Au/Cu/POAP/GOD biosensor has lower detection limit
(0.01 mM), larger maximum current density (0.45 mA cm−2 )
and higher sensitivity (12.6 mA M−1 cm−2 ) due to the exis-
tence of Cu nanoparticles. Moreover, the Au/Cu/POAP/GOD
electrode also exhibits good selectivity, large response cur-
rent, fast amperometric response, good reproducibility and
excellent stability.
Acknowledgements
This project was financially supported by the Scientific
Research Foundation for the Returned Overseas Chinese
Scholars, State Education Ministry (2001-498); the National
Natural Science Foundation of China (No. 20275009) and
the foundation of the Ministry of Science and Technology
of China (2001-51).
References
[1] M.C. Rodriguez, G.A. Rivas, Amperometric glucose biosensor based
on the deposition of copper and glucose oxidase onto glassy carbon
transducer, Anal. Lett. 33 (2000) 2373–2388.
[2] S. Yabuki, F. Mizutani, Y. Satao, Y. Hirata, Immobilization of
polyglutamate–glucose oxidase onto a cysteamine-modified gold
electrode, Sens. Actuators, B 91 (2003) 187–190.
[3] K. Ban, T. Ueki, Y. Tamada, T. Saito, S.I. Imabayashi, M. Watanabe,
Electrical communication between glucose oxidase and electrodes
mediated by phenothiazine-labeled poly(ethylene oxide) bonded to
lysine residues on the enzyme surface, Anal. Chem. 75 (2003) 910–
917.
73
[4] Y.M. Uand, T.C. Chou, Fabrication of glucose oxidase/polypyrrole
biosensor by galvanostatic method in various pH aqueous solutions,
Biosens. Bioelectron. 19 (2003) 141–147.
[5] C.W. Chuang, J.S. Shih, Preparation and application of immobilized
C60-glucose oxidase enzyme in fullerene C60-coated piezoelectric
quartz crystal glucose sensor, Sens. Actuators B 81 (2001) 1–8.
[6] H. Yang, T.D. Chung, Y.T. Kim, C.A. Choi, C.H. Jun, H.C. Kim,
Glucose sensor using a microfabricated electrode and electropoly-
merized bilayer films, Biosens. Bioelectron. 17 (2002) 251–259.
[7] A.A. Karyakin, E.E. Karyakina, L. Gorton, Prussian-Blue-based am-
perometric biosensors in flow-injection analysis, Talanta 43 (1996)
1597–1606.
[8] R. Garjonyte, A. Malinauskas, Glucose biosensor based on glu-
cose oxidase immobilized in electropolymerized polypyrrole and
poly(o-phenylenediamine) films on a Prussian Blue-modified elec-
trode, Sens. Actuators B 63 (2000) 122–128.
[9] D.W. Pan, J.H. Chen, L.H. Nie, W.Y. Tao, S.Z. Yao, An amperometric
glucose biosensor based on poly(o-aminophenol) and Prussian Blue
films at platinum electrode, Anal. Biochem. 324 (2004) 115–122.
[10] R. Garjonyte, A. Malinauskas, Amperometric glucose biosensor
based on glucose oxidase immobilized in poly(o-phenylenediamine)
layer, Sens. Actuators B 56 (1999) 85–92.
[11] P.N. Bartlett, J.M. Cooper, A review of the immobilization of en-
zymes in electropolymerized films, J. Electroanal. Chem. 362 (1993)
1–12.
[12] M. Gerard, A. Chaubey, B.D. Malhotra, Application of conducting
polymers to biosensors, Biosens. Bioelectron. 17 (2002) 345–359.
[13] C. Malitesta, F. Palmisano, L. Torsi, P.G. Zambonin, Glucose
fast-response amperometric sensor based on glucose oxidase immo-
bilized in an electropolymerized poly(o-phenylenediamine), Anal.
Chem. 62 (1990) 2735–2740.
[14] J.J. Xu, H.Y. Chen, Amperometric glucose sensor based on
glucose oxidase immobilized in electrochemically generated
poly(ethacridine), Anal. Chim. Acta 423 (2000) 101–106.
[15] C.A. Groom, J.H.T. Luong, Improvement of the selectivity of am-
perometric biosensors by using a permselective electropolymerized
film, Anal. Lett. 26 (1993) 1383–1389.
[16] P.N. Bartlett, D.J. Caruana, Electrochemical immobilization of en-
zymes. Part V∗ . Microelectrodes for the detection of glucose based
on glucose oxidase immobilized in a poly(phenol) film, Analyst 117
(1992) 1287–1292.
[17] Z.N. Zhang, H.Y. Liu, J.Q. Deng, A glucose biosensor based on im-
mobilization of glucose oxidase in electropolymerized o-aminophenol
film on platinized glassy carbon electrode, Anal. Chem. 68 (1996)
1632–1638.
[18] Y. Nakabayashi, M. Wakuda, H. Imai, Amperometric glucose sensors
fabricated by electrochemical polymerization of phenols on carbon
paste electrodes containing ferrocene as electron transfer mediator,
Anal. Sci. 14 (1998) 1069–1076.
[19] Y. Nakabayashi, H. Yoshikawa, Amperometric biosensors for sensing
of hydrogen peroxide based on electron transfer between horseradish
peroxide and ferrocene as a mediator, Anal. Sci. 16 (2000) 609–613.
[20] C. Barbero, J.J. Silber, L. Sereno, Formation of a novel electroac-
tive film by electropolymerization of ortho-aminopheol. Study of its
chemical structure and formation mechanism. Electropolymerization
of analogous compounds, J. Electroanal. Chem. 263 (1989) 333–352.
[21] M.A.V. Garcı́a, P.T. Blanco, A. Ivaska, A poly(o-aminopheol) mod-
ified electrode as an amperometric hydrogen peroxide biosensor,
Electrochim. Acta 43 (1998) 3533–3539.
[22] E. Miland, A.J. Mı́randa Ordieres, P.T. Blanco, Poly(o-aminophenol)-
modified bienzyme carbon paste electrode for the detection of uric
acid, Talanta 43 (1996) 785–796.
[23] D.W. Pan, J.H. Chen, L.H. Nie, W.Y. Tao, S.Z. Yao, Amperometric
glucose biosensor based on immobilization of glucose oxidase in
electropolymerized o-aminophenol film at Prussian Blue-modified
platinum electrode, Electrochim. Acta 49 (2004) 795–801.
74 D. Pan et al. / Sensors and Actuators B 104 (2005) 68–74
[24] I.G. Casella, M. Gatta, M.R. Guascito, T.R.I. Cataldi, Highly-
dispersed copper microparticles on the active gold substrate as an
amperometric sensor for glucose, Anal. Chim. Acta 357 (1997) 63–
71.
[25] G.A. Ragoisha, A.S. Bondarenko, Potentiodynamic electrochemical
impedance spectroscopy. Copper underpotential deposition on gold,
Electrochem. Commun. 5 (2003) 392–395.
[26] J.M. Ortega, Electrodeposition of copper on poly(o-aminophenol)
modified platinum electrode, Thin Solid Film 360 (2000) 159–165.
[27] M. Somasundrum, K. Kirtikara, M. Tanticharoen, Amperometric de-
termination of hydrogen peroxide by direct and catalytic reduction
at a copper electrode, Anal. Chim. Acta 319 (1996) 59–70.
[28] J.F. Zhou, J.J. Yang, Z.J. Zhang, W.M. Liu, Q.J. Xue, Study on the
structure and tribological properties of surface-modified Cu nanopar-
ticles, Mater. Res. Bull. 34 (1999) 1361–1367.
[29] S.S. Joshi, S.F. Patil, V. Iyer, S. Mahumuni, Radiation induced
synthesis and characterization of copper nanoparticles, Nanostruct.
Mater. 10 (1998) 1135–1144.
[30] S.L. Mu, H.G. Xue, B.D. Qian, Bioelectrochemical responses of the
polyaniline glucose oxidase electrode, J. Electrochem. Chem. 304
(1991) 7–16.
[31] H. Zheng, H.G. Xue, Y.F. Zhang, Z.Q. Shen, A glucose biosen-
sor based on microporous polyacrylonitrile synthesized by single
rare-earth catalyst, Biosens. Bioelectron. 17 (2002) 541–545.
Biographies
Jinhua Chen obtained Doctor’s degree from College of Chemistry and
Chemical Engineering, Hunan University, PR China in 1997. He is
presently employed as professor in College of Chemistry and Chemical
Engineering, Hunan University, PR China. He is mainly engaged in the
research of chemo/biosensors, carbon nanotube and applied electrochem-
istry.
Dawei Pan obtained Bachelor and Master’s degree from College of Chem-
istry and Chemical Engineering, Hunan University, PR China. He is
presently a PhD student in College of Chemistry and Chemical Engineer-
ing, Hunan University, PR China. He is mainly engaged in the research
of biosensors and bioelectrochemistry.
Shouzhuo Yao is presently employed as a professor in College of Chem-
istry and Chemical Engineering, Hunan University, PR China. He is
mainly engaged in the research of analytical chemistry, piezoelectrical
sensors and molecular imprinting polymer.
Lihua Nie is presently employed as a senior engineer in College of
Chemistry and Chemical Engineering, Hunan University, PR China. She
is mainly engaged in the research of analytical chemistry and molecular
imprinting polymer.
Jianjun Xia is a postgraduate student of College of Chemistry and Chem-
ical Engineering, Hunan University, PR China. He is mainly engaged in
the research of bioelectrochemistry.
Wenyan Tao is a PhD student in College of Chemistry and Chemical
Engineering, Hunan University, PR China. She is mainly engaged in the
research of biosensors and bioelectrochemistry.