Colloids and Surfaces A: Physicochem. Eng.

Aspects 456 (2014) 146–152

Contents lists available at ScienceDirect

Colloids and Surfaces A: Physicochemical and

Engineering Aspects
journal homepage: www.elsevier.com/locate/colsurfa

Direct electrodeposition of reduced graphene oxide on carbon fiber

electrode for simultaneous determination of ascorbic acid, dopamine
and uric acid
Beibei Yang a , Huiwen Wang a , Jiao Du a , Yunzhi Fu b,∗ , Ping Yang a , Yukou Du a,∗
a
College of Chemistry, Chemical Engineering and Materials Science, Soochow University, Suzhou 215123, PR China
b
Department of Materials and Chemical Engineering, Hainan University, Haikou 570228, PR China

h i g h l i g h t s g r a p h i c a l a b s t r a c t

• The ErGO/CFE was fabricated by a

simple electrodeposition method.
• The ErGO/CFE shows high electron
transfer kinetics in CV response.
• The ErGO/CFE shows high sensitiv-
ity and selectivity for simultaneous
determination of AA, DA and UA.

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

Article history: The electrochemically reduced graphene oxide (ErGO) modified carbon fiber electrode (CFE) was fabri-
Received 25 March 2014 cated by a simple electrodeposition method. Cyclic voltammetry (CV) and differential pulse voltammetry
Received in revised form 8 May 2014 (DPV) had been used to investigate the electrochemical behavior of ErGO/CFE toward ascorbic acid (AA),
Accepted 14 May 2014
dopamine (DA) and uric acid (UA). Compared to the bare CFE, the ErGO/CFE showed higher catalytic
Available online 21 May 2014
activity toward electrochemical oxidation of AA, DA and UA. In the CV curves, three well and distinct
peaks with large peak potential separation of 116 mV, 167 mV and 283 mV between AA–DA, DA–UA and
Keywords:
AA–UA were observed. In the DPV curves, the corresponding peak potential separations were 198 mV,
Electrodeposition
ErGO 163 mV and 361 mV between AA–DA, DA–UA and AA–UA also observed. DPV was used as the analyti-
Ascorbic acid cal technique to acquire the linear calibration curves for AA, DA and UA with the concentration ranges
Dopamine of 8–2016.45 ␮M, 1.5–224.82 ␮M, 6–899.3 ␮M, respectively, in the individual detection of each com-
Uric acid ponent. The detection limits were 4.5 ␮M, 0.77 ␮M and 2.23 ␮M for AA, DA and UA indicating that the
ErGO/CFE can be achieved with high sensitivity and selectivity for simultaneous determination of these
three biomolecules.
© 2014 Elsevier B.V. All rights reserved.

1. Introduction commonly used in large scale as an antioxidant in food, animal

Ascorbic acid (AA), dopamine (DA) and uric acid (UA) play
important roles in physiological function of organisms. AA is
∗ Corresponding authors. Tel.: +86 512 65880089; fax: +86 512 65880089.
E-mail addresses: yzhfu@hainu.edu.cn (Y. Fu), duyk@suda.edu.cn (Y. Du).
feed, pharmaceutical formulations and cosmetic applications [1].
Meanwhile AA has direct participation in many biological reactions
and its deficiency can be related to symptom of some diseases, e.g.
scurvy [2]. It also has been used for prevention and treatment of
common cold, mental illness, infertility, cancer and AIDS [3]. DA is
an important neurotransmitter that belongs to the catecholamine
family and has been given extensive attention in clinical research

http://dx.doi.org/10.1016/j.colsurfa.2014.05.029
0927-7757/© 2014 Elsevier B.V. All rights reserved.
 B. Yang et al. / Colloids and Surfaces A: Physicochem. Eng. Aspects 456 (2014) 146–152
due to its involvement in motor and cognitive function [4]. Abnor-
mal DA concentration in the brain may lead to serious disease, such
as Parkinson’s disease and addiction [5,6]. UA is the final oxidation
product of urine metabolism and is excreted in urine [7]. Abnormal
concentration of UA in human body is symptom of several diseases
such as gout, hyperuricemia and Lesch–Nyhan syndrome [8,9].
Therefore, the sensitive and reliable determination of AA, DA and
UA is an important topic not only in the field of biomedical chem-
istry but also for diagnostic and pathological research. In present,
electrochemical methods have been extensively used for the deter-
mination of these three molecules. However, as we know, the three
biological molecules oxidation potentials are almost the same at
traditional electrodes which results in severely overlapped voltam-
metric response [10]. It is difficult to simultaneously determine
them at the traditional electrodes, so various chemically modified
electrodes have been developed.
Graphene, a one-atom-thick sp2 -bond carbon sheet recently
has been attracted extensive interest because of its ideal two-
dimensional structure [11], unique electronic properties [12],
thermal properties [13], mechanical properties [14] and opti-
cal properties [15]. It is always applying in many fields such
as nanoelectronics, nanoelectromechanical devices, nanocompos-
ites, sensors, ultracapacitors, solar cells and liquid crystal devices
[16–18]. Furthermore, graphene oxide (GO) and reduced graphene
oxide (RGO) have also been used for modified electrodes. GO
with single-layered is an intriguing nanomaterial with tremendous
potential for electronic applications. Many reports are related to the
application of graphene oxide based materials in supercapacitors
[19]. Compared to graphene oxide, the reduced graphene oxide has
better conductivity. Studies indicate that biological activity of the
RGO modified electrode shows a better electrochemical response
[20]. Wang et al. reviewed the application of RGO in electrochem-
ical catalysis [21], Chen and Tang also reviewed the application of
RGO in electrochemical sensor [22]. To the best of our knowledge,
there are no reports which use the electrodeposition method to
obtain RGO for simultaneous determination of AA, DA and UA.
In this work, graphene oxide was directly electrodeposited
at a constant potential on the carbon fiber electrode to obtain
the reduced graphene oxide modified electrode (ErGO/CFE). The
obtained ErGO/CFE was used for simultaneous determination of
AA, UA and DA. The basic characteristics of the ErGO electrode
were studied in details. Cyclic voltammetry and differential pulse
voltammetry were employed to investigate the electrochemical
behaviors of AA, DA and UA at the proposed electrode. The reusabil-
ity and stability of the ErGO electrode were also studied.
2. Experimental
2.1. Reagents
Graphite powder, disodium hydrogen phosphate, sodium dihy-
drogen phosphate, lithium perchlorate, potassium ferrocyanide,
potassium ferricyanide and potassium chloride were purchased
from Sinopharm Chemicals Reagent Co., Ltd., China. Ascorbic acid,
dopamine and uric acid were purchased from Acros Organics. All
these chemicals were of analytical reagent grade and used as
received. The solutions of AA, DA and UA were freshly prepared
with 0.1 M phosphate buffer solutions (PBS; pH 7.0). The double-
distilled water was used throughout all the experiments.
2.2. Apparatus
The morphology of obtained electrode was studied by scan-
ning electron microscope (SEM) (S-4700, Hitachi High Technologies
Corporation, Japan). Cyclic voltammetry and differential pulse
147
voltammetry measurements were carried out using a CHI650D
electrochemical workstation (Shanghai Chenhua Instrument Plant,
China) with a standard three-electrode. The CFE (360 ␮m diameter)
(CeTech Co., Ltd.) and its modified electrode were used as working
electrodes. CFEs were rinsed thoroughly with ethanol and water
by ultrasonication each for 15 min before experiments. Platinum
foil with an area of 1 cm × 1 cm and a saturated calomel elec-
trode (SCE) were used as counter electrode and reference electrode,
respectively. The areas of working electrode and counter electrode
immersed in the solution were 1.2 cm × 360 ␮m and 0.5 cm × 1 cm,
respectively. All the experiments were carried out at room temper-
ature.
2.3. Preparation of graphene oxide
Graphene oxide (GO) was synthesized from graphite by modi-
fied Hummers method [23]. Firstly, 0.5 g NaNO3 and 1.0 g graphite
powder were added into a 24 mL H2 SO4 solution (0 ◦ C). Under
vigorous agitation, 3.0 g KMnO4 was added slowly to keep the
temperature of the suspension lower than 20 ◦ C. Successively, the
reaction system was transferred to a 35 ◦ C water bath and stirred for
60 min. Secondly, 46 mL of double-distilled water was mixed into
the above solution and further stirred at 98 ◦ C for 15 min. Addi-
tional 140 mL of double-distilled water was added and followed by
a slow addition of 10 mL H2 O2 (30%) turning the reaction to stop.
After the above reactions, the final product was washed by HCl (5%)
until sulfate ions could not be detected with BaCl2 . The final prod-
uct was then dried in vacuum for 12 h at 40 ◦ C. Lastly, 60 mg the
product GO was dispersed into 10 mL double-distilled water under
ultrasonic homogenizer for 120 min so that the impurities in the
GO solution were centrifuged. After the GO solution was purified
by centrifugal machine for 15 min, we got target GO solution with
the concentration was estimated to be 3 mg/mL.
2.4. Preparation of the ErGO/CFE electrode
The electrochemically reduced graphene oxide (ErGO) modified
electrode as prepared by electrolyzing the 3 mg/mL GO aqueous
suspension containing 0.1 M lithium perchlorate on the CFE at a
consistent potential of −1.2 V for 5000 s (shown in Scheme 1). After
electrodeposition, the ErGO electrode was washed with double-
distilled water and then immersed in double-distilled water for
30 min to remove the residual GO absorbed on the electrode.
3. Results and discussion
3.1. Basic characterizations of ErGO
The morphology of CFE and obtained ErGO/CFE was investigated
by SEM. Fig. 1 shows that there are obvious differences between
the two electrodes. From Fig. 1(A), we can observe a quite smooth
surface of CFE, while in Fig. 1(B) and (C) the surface of ErGO/CFE
is wholly covered with three-dimensional structures. How do the
three-dimensional structures form? It is apparently that graphene
oxide was reduced at the consistent potential. With an increasing
electrodeposition time, the reduced graphene oxide sheets on CFE
surface would increase. The reduced graphene oxide sheets inter-
connected with each other and as a result, the three-dimensional
structures formed on the surface of ErGO/CFE. It can be believed that
the three-dimensional structures enlarge the surface area of the
ErGO/CFE which could provide a high area for the affinity adsorp-
tion of AA, DA and UA.
Fig. 2 compares the cyclic voltammetric (CV) responses at the
two electrodes in 2.5 mM Fe (CN)6 3−/4− + 0.1 M KCl solution. The CV
curve of bare CFE shows a pair of poor redox peaks, suggesting the
sluggish electron transfer at the interface of the carbon fiber. While
148 B. Yang et al. / Colloids and Surfaces A: Physicochem. Eng. Aspects 456 (2014) 146–152

Scheme 1. Illustration for the fabrication of ErGO/CFE.

in the CV response of ErGO/CFE, well-defined peaks with a small
peak potential separation (Ep ) of 70 mV are observed, suggesting
that the rapid electron transfer rate was obtained on the ErGO/CFE.
The enhanced electron transfer kinetics on the ErGO/CFE might be
attributed to the high electric conductivity of graphene [24] and the
plenty of reduced graphene oxide sheets on the electrode surface.
3.2. Cyclic voltammetric detection of AA, DA, UA and their ternary
mixture
Fig. 3 shows the cyclic voltammetric responses at bare CFE
and ErGO/CFE in 0.1 M PBS solution (pH 7.0) containing 8 mM
AA, 0.5 mM DA and 2 mM UA, respectively. At the bare CFE,
AA (Fig. 3A) and UA (Fig. 3C) show irreversible electrochemical
behaviors with relatively weak redox currents. For DA (Fig. 3B),
the oxidation and reduction peaks appear at 235 mV and 98 mV,
respectively, and the peak potential separation (Ep ) is about
133 mV. It is clear that the electrochemical reactions of these
three biological molecules at the bare CFE are irreversible and
undergo sluggish electron transfer kinetic processes. In com-
parison to the bare CFE, the electrochemical reactions of these
three biological molecules at the ErGO/CFE exhibit a significant
enhancement. The oxidation peak–reduction peak potential sepa-
rations of the three biological molecules are significantly narrowed.

Fig. 1. SEM images of the bare CFE (A), ErGO/CFE (B) and the enlarged image of ErGO/CFE (C).
 B. Yang et al. / Colloids and Surfaces A: Physicochem. Eng. Aspects 456 (2014) 146–152 149

Fig. 2. CV responses of CFE and ErGO/CFE in 2.5 mM Fe (CN)6 3−/4− + 0.1 M KCl solu- Fig. 4. CV responses of the mixture containing 9.6 mM AA, 0.6 mM DA and 2.4 mM
tion at 50 mV/s. UA in 0.1 M PBS solution (pH 7.0) on CFE and ErGO/CFE at 50 mV/s.

For AA, the oxidation peak potential negatively shifts about 261 mV
comparing with the potential at bare CFE, so large peak potential
shift activates the hydroxyl in the furan nucleus of AA, which may be
caused by the remaining carboxyl of ErGO and formation of hydro-
gen bonds between lactone bonds of AA [25]. That shift makes it
possible to determine these three biological molecules at the same
time. Moreover, the oxidation peak current of AA is 62 ␮A, which
is about 2 times higher than that at bare CFE. For DA the oxidation
and reduction peaks are located at 224 mV and 176 mV showing
a smaller peak-to-peak separation than that at bare CFE. The Ep
is 48 mV and the oxidation peak current is 4.5 times higher than
that at bare CFE suggesting the excellent electrocatalytic activity of
ErGO toward DA. The similar electrocatalytic activity for UA is also
found, which occurs with an increased peak current (8.2 times)
and a small Ep (55 mV) compared with bare CFE, indicating good
electron transfer promotion ability of ErGO modified material.
Fig. 4 shows the CV responses at bare CFE and ErGO/CFE in a
ternary mixture with 9.6 mM AA, 0.6 mM DA and 2.4 mM UA in
0.1 M PBS solution (pH 7.0). At the bare CFE only a broad and over-
lapped oxidation peak is obtained at 413 mV and the potentials of
AA, DA and UA are indistinguishable so that simultaneous determi-
nation of these biological molecules is impossible. However, at the
ErGO/CFE, three sharp and well defined oxidation peaks with larger
peak separation potential and higher peak currents correspond-
ing to the oxidation of AA, DA and UA appear. The three oxidation
peaks for AA, DA and UA are well resolved at 83 mV, 199 mV and

Fig. 3. CV responses of (A) AA (8 mM), (B) DA (0.5 mM) and (C) UA (2 mM) in 0.1 M PBS solution (pH 7.0) on CFE and ErGO/CFE at 50 mV/s, respectively.
150 B. Yang et al. / Colloids and Surfaces A: Physicochem. Eng. Aspects 456 (2014) 146–152

Fig. 5. CV responses of (A) 8 mM AA, (B) 0.5 mM DA and (C) 2 mM UA in 0.1 M PBS solution (pH 7.0) with different scan rates at ErGO/CFE. Scan rate in the range from 20 mV/s
to 200 mV/s. Insets: the relation between oxidation currents versus the scan rates of AA, DA and UA, respectively.

366 mV, respectively, with the peak separation potential is 116 mV
(AA–DA), 167 mV (DA–UA) and 283 mV (AA–UA). Meanwhile, we
can observe the oxidation peak potential of these three biological
molecules shift compared with those in their individual detection.
It may be caused by the change of interface oxidation properties
which are produced when the products of AA, DA and UA molecules
adsorbed on the surface of the ErGO/CFE [26]. All the CV responses
can demonstrate that the ErGO modified electrode possess excel-
lent electrocatalytic activities towards the oxidation of AA, DA and
UA, which can be attributed to its unique structural features and
excellent electrochemical properties.
3.3. Effect of scan rate on the electrochemical oxidation of AA, DA
and UA
3.4. Simultaneous determination of AA, DA and UA by DPV
Determination of AA, DA and UA in their mixture was per-
formed by employing the DPV response at bare CFE and ErGO/CFE
in 0.1 M PBS (pH 7.0) solution containing 9.6 mM AA, 0.6 mM DA
and 2.4 mM UA. As shown in Fig. 6, at the bare CFE, the elec-
trooxidation of these three biological molecules presents only two
indistinct peaks so that determination of the individual concentra-
tion of these molecules in their mixture is impossible. In contrast,
at the ErGO/CFE three well defined oxidation peaks were observed
at −15 mV, 183 mV and 346 mV for AA, DA and UA respectively,
with higher (approximately 2.6 times) oxidation peak currents
appeared. The large peak separations 198 mV, 163 mV and 361 mV

As we know, investigating the effect of scan rate on the oxidation

peak current and peak potential can evaluate the kinetics of elec-
trode reaction [27]. Fig. 5 shows the CV responses at the ErGO/CFE
with different scan rates in 0.1 M PBS solution (pH 7.0) containing
8 mM AA, 0.5 mM DA and 2 mM UA, respectively. When the scan
rate changes from 20 mV/s to 200 mV/s, the oxidation peak currents
of AA (Fig. 5A), DA (Fig. 5B) and UA (Fig. 5C) increase with the expan-
sion of scan rate, while their oxidation peak potentials gradually
shift to positive values. Moreover, the relationship between oxida-
tion peak currents and the scan rates is linear. The linear equations
are expressed as follows:
Ipa (␮A) = 0.0455 (mV/s) − 0.6018 (AA); Ipa (␮A) = 0.382
(mV/s) + 3.134 (DA); Ipa (␮A) = 0.2203 (mV/s) + 2.8292 (UA) with
correlation coefficient of R2 = 0.9933, 0.9977, 0.9878, respectively.
The linear relationships indicate that the three biological molecules
are all dominated by the surface adsorption-controlled process Fig. 6. DPV responses of the mixture containing 9.6 mM AA, 0.6 mM DA and 2.4 mM
[28]. UA in 0.1 M PBS solution (pH 7.0) on CFE and ErGO/CFE at 50 mV pulse amplitude.
 B. Yang et al. / Colloids and Surfaces A: Physicochem. Eng. Aspects 456 (2014) 146–152
Fig. 7. DPV responses of (A) 8–2016.45 ␮M AA, (B) 1.5–224.82 ␮M DA and (C) 6-899.3 ␮M UA in 0.1 M PBS solution (pH 7.0) on ErGO/CFE at 50 mV pulse amplitude. Insets:
the relation between oxidation current versus the concentration of AA, DA and UA, respectively.
for AA–DA, DA–UA and AA–UA allow the simultaneous determina-
tion of AA, DA and UA in their mixture, indicating that the ErGO/CFE
in this work possesses good selectivity and sensitivity toward AA,
DA and UA admixture.
3.5. Individual determination of AA, DA and UA by DPV
Compared to the above CV response, DPV response has better
resolution and higher sensitivity, so it was used to determine indi-
vidual AA, DA and UA at the ErGO electrode in order to increase
the detection sensitivity and selectivity [29,30]. Fig. 7 shows the
DPV responses of the concentration AA from 8 ␮M to 2016.45 ␮M,
DA from 1.5 ␮M to 224.82 ␮M and UA from 6 ␮M to 899.3 ␮M. The
peak potential is −76 mV, 124 mV and 233 mV for AA (Fig. 7A), DA
(Fig. 7B) and UA (Fig. 7C), respectively, and they have almost no shift
when the concentrations of these biological molecules increase.
Different from the potentials, their peak currents gradually increase
as the concentrations increase. The linear relationships between
their peak currents and concentrations are Ipa (␮A) = 0.0667cAA
(␮M) − 3.5669 (AA); Ipa (␮A) = 0.1024cDA (␮M) − 1.48 (DA); Ipa
(␮A) = 0.0379cUA (␮M) − 1.1685 (UA) with correlation coefficient of
R2 = 0.9984, 0.9938, 0.9973, respectively. Meanwhile, the detection
limits were 4.5 ␮M (AA), 0.77 ␮M (DA) and 2.23 ␮M (UA) indicating
that the ErGO/CFE can be achieved with high sensitivity and selec-
tivity for simultaneous determination of these three biomolecules.
3.6. Reusability and stability of ErGO/CFE
The reusability of ErGO/CFE was estimated by measuring the
DPV responses of the mixture containing 1.32 mM AA, 0.12 mM DA
and 0.48 mM UA in 0.1 M PBS solution (pH 7.0) with five electrodes
produced under the same condition. The relative standard devi-
ations of the oxidation peak currents are 3.28%, 1.93% and 2.26%
151
for AA, DA and UA, respectively. The good reusability means that
ErGO/CFE is promising for the simultaneous determination of these
three molecules in analytical application. The stability of ErGO/CFE
also was checked by performing the modified electrode which was
stored a week at 25 ◦ C before the experiment. The peak currents
intensity only decreased 4.6%, 3.1% and 3.8% for AA, DA and UA,
respectively. These results above indicate that the modified elec-
trode shows a well reusability and good stability.
4. Conclusions
The present work shows that electrodeposition and electrore-
duction of graphene oxide at a constant potential has been
successfully performed on carbon fiber electrode, which is used
for simultaneous determination of AA, DA and UA. With three-
dimensional structures consist of reduced graphene oxide sheets,
the obtained ErGO/CFE exhibited remarkable increase in the elec-
tron transfer kinetics toward the oxidation reaction of these three
molecules. In the CV or DPV responses large peak potential sep-
aration and high peak current could be observed at the obtained
electrodes. Therefore, these biomolecules can be simultaneously
determined from their mixture solution with high sensitivity. In
addition, the ErGO/CFE also showed a well reusability and a good
stability in this experiment. The present work demonstrates that
the electrochemically reduced graphene oxide is a promising candi-
date electrode material for bioelectrochemical devices in the future.
Acknowledgments
This work was supported by the National Natural Science Foun-
dation of China (Grant nos. 51373111, 51073114 and 20933007),
Suzhou Nano-project (ZXG2012022), the Opening Project of Xin-
jiang Key Laboratory of Electronic Information Materials and
152 B. Yang et al. / Colloids and Surfaces A: Physicochem. Eng. Aspects 456 (2014) 146–152
Devices (XJYS0901-2010-01), the Academic Award for Young
Graduate Scholar of Soochow University, the Priority Academic
Program Development of Jiangsu Higher Education Institutions
(PAPD).
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