Composites Science and Technology 125 (2016) 71e79

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Composites Science and Technology

journal homepage: http://www.elsevier.com/locate/compscitech

Characterization protocol to improve the electroanalytical response of

grapheneepolymer nanocomposite sensors
Jose Mun~ oz a, Lorcan J. Brennan b, Francisco Ce
spedes a, Yurii K. Gun'ko b, c,
Mireia Baeza a, *
a
Departament de Química, Facultat de Ciencies, Edifici C-Nord, Universitat Autonoma de Barcelona, 08193 Cerdanyola del Vall
es (Bellaterra), Barcelona,
Spain
b
School of Chemistry and CRANN Institute, Trinity College Dublin, Dublin 2, Ireland
c
ITMO University, 197101 Saint Petersburg, Russia

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

Article history: This work is focused on the application of advanced characterization techniques for the development of
Received 3 June 2015 optimized electrochemical sensors based on grapheneepolymer nanocomposites. Reduced graphene
Received in revised form oxide (rGO) was synthesized and used as conducting nanofiller material in nanocomposite electrodes
19 January 2016
based on epoxy resin. A series of rGO/epoxy nanocomposite electrodes were fabricated containing 8%
Accepted 21 January 2016
e20% of nanofiller material. Composition ratios have been studied using percolation theory and char-
Available online 22 January 2016
acterized by different electrochemical techniques, demonstrating that an optimization of the rGO/epoxy
composition ratio is mandatory to enhance electrochemical performance. The potential of this approach
Keywords:
A. Nano composites
in terms of electroanalytical response has been demonstrated by means of the amperometric detection of
A. Polymer-matrix composites (PMCs) ascorbic acid, which was used as a model analyte. Finally, different morphological experiments were also
B. Electrical properties carried out in order to verify the electrochemical and electroanalytical enhancement obtained for the
B. Impact behaviour optimized-nanocomposite sensors.
B. Transport properties © 2016 Elsevier Ltd. All rights reserved.

1. Introduction rGO exhibit a higher sensitivity and electrocatalytic activity than

During the past few decades the electrochemical properties of
different carbon paste electrodes were studied in detail [1]. Carbon
composites for electrochemical applications were introduced for
first time in 1958 by Adams [2]. Recently large research interest has
focused on graphene-based nanocomposite materials [3e5]. Due to
their good levels of conductivity, thermal stability and excellent
mechanical strength, graphene and its derivatives are important
nanofiller materials for polymer nanocomposites. Regarding to
their electrochemical properties, graphene-based nanocomposite
materials possess important characteristics, such as large specific
surface area, excellent conductivity and availability for surface
functionalization [6]. As another important member of the gra-
phene family, graphene oxide (GO) and reduced graphene oxide
(rGO) are more electrochemically active when compared to pristine
graphene owing to the oxygen-containing reactive sites at edges
and in the basal plan [7]. In spite of this fact, electrodes based on
* Corresponding author.
E-mail address: mariadelmar.baeza@uab.cat (M. Baeza).
most GO-based electrodes, due to the higher conductivities asso-
ciated with the increased level of in-plane sp2 hybridization [8,9].
The properties and performances of graphene-polymer nano-
composites not only depends on the quality of the graphene filler
and polymer matrix, but performance will also depend on the
dispersity of the filler and the ratio of filler to the matrix [10]. The
electrical conductivity is one of the most important changes when
graphene-based nanocomposites are prepared, following normally
a percolation behaviour [11]. However, depending on the conduc-
tive load, nanocomposites can behave as microelectrode arrays
which are known to provide efficient mass transport of the elec-
troactive species due to radial diffusion on the spaced carbon par-
ticles [12]. This fact favours the sensitive electroanalysis of a variety
of reagents. Under this context, another point of consideration is
the optimization of the carbon loading in the nanocomposite ma-
terials for improving their electrochemical properties and analyt-
ical applications.
To date, many electrochemical groups have focused their
attention on the optimization of the nanocomposite proportions
with the aim of maximizing the conductive material loading

http://dx.doi.org/10.1016/j.compscitech.2016.01.018
0266-3538/© 2016 Elsevier Ltd. All rights reserved.
72 ~oz et al. / Composites Science and Technology 125 (2016) 71e79
J. Mun
without losing its physical and mechanical stability, in order to
confer the lowest bulk resistivity [13] and [14]. However, high
carbon loadings can increase the background current and smear the
Faradaic signal response, especially when the electroactive species
are present in low concentration. From an analytical point of view,
the requirements to achieve good sensor performance are high
sensitivity, rapid response time and low limit of detection. These
requirements are related to some of the materials physical pa-
rameters such as material resistivity, heterogeneous electron
transfer rate and double-layer capacitance [15].
Accordingly, the goal of this work is based on developing opti-
mized graphene-based epoxy nanocomposite sensors for their use
in electroanalysis. For this aim, rGO was used as conducting
nanofiller material and dispersed through an insulating polymer
matrix (epoxy resin) to prepare different compositions of nano-
composites. Following that, different electrical and electrochemical
tools were systematically carried out to achieve the best electrode
performance, which includes low resistances and high signal-to-
noise ratio. Additionally, morphological and topographical charac-
terizations were also carried out in order to gain a greater insight
into the surface properties of the electrodes. Finally, the benefits of
using this approach in terms of analytical performance have been
demonstrated by the successful detection of ascorbic acid, which
was used as a model analyte, by hydrodynamic amperometry.
2. Experimental
2.1. Chemicals and reagents
Graphene Oxide (GO) was synthesized from flaked graphite
(Alfa Aesar, Karlsruhe, Germany) using the well documented
Hummers method [16]. Reduced Graphene Oxide (rGO) was ob-
tained through the reduction of GO using ascorbic acid as a
reducing agent [17]. Epotek H77A and its corresponding hardener
Epotek H77B, from Epoxy Technology (Billerica, MA, USA), were
used as the polymeric matrix. All solutions were prepared using
deionised water from a Milli-Q system (Millipore, Billerica, MA,
USA). Potassium ferricyanide/ferrocyanide (99.8%), ascorbic acid
(99.5%), potassium nitrate (99.0%), potassium chloride (99.5%), ni-
tric acid (65%), potassium permanganate (99.99%), sulphuric acid
(95e98%) and hydrochloric acid (30e35%) were purchased from
SigmaeAldrich (St. Louis, MO, USA).
2.2. Apparatus and procedure
Electrochemical experiments were carried out by Cyclic Vol-
tammetry (CV) and Electrochemical Impedance Spectroscpy (EIS),
using a pontentiostat/galvanostat Autolab system (PGSTAT 30 and
FRA boards, Eco Chemie, Utrech, The Netherlands) with a three-
electrode configuration. The system was run on a PC using GPES
and FRA 4.9 software. An AgCl covered silver wire and a platinum-
based electrode 52-671 (Crison Instruments, Alella, Barcelona,
Spain) were used as reference and auxiliary electrodes, respec-
tively. The graphene/epoxy nanocomposites were employed as
working electrodes. The measurements were made in a 10.0 mL of
0.1 M KCl containing 0.01 M K3[Fe(CN)6/K4[Fe(CN)6]. All the ex-
periments were performed at room temperature (25
C). Cyclic
voltammograms were carried out using a scan rate of 50 mV s1. For
impedance measurements, a frequency range of 100 kHze0.1 Hz
was employed at redox equilibrium potential, which was previ-
ously obtained by CV. The signal amplitude to perturb the system
was 10 mV and the equilibrium time was 15 s.
Electroanalytical experiments were carried out in a 10.0 mL
solution of 0.01 M HNO3/KNO3, which was used as a background
electrolyte. The experiments were made using a three-electrode
configuration. A double junction reference electrode Ag/AgCl
Orion 900200 (Thermo Electron Corporation, Beverly, MA, USA)
and a platinum-based electrode 52-671 (Crison Instruments, Alella,
Barcelona, Spain) were used as reference and auxiliary respectively;
graphene/epoxy nanocomposites were used as different working
electrodes. Amperometric measurements were performed using an
amperimeter LC-4C (Bioanalytical Systems, Inc., West Lafayette, IN,
USA). Amperometric detection was performed under stirring con-
ditions at 600 mV vs. Ag/AgCl fixed potential. Ascorbic acid was
used as a model analyte for evaluating the electroanalytical char-
acteristics of the sensors. A freshly prepared 0.01 M ascorbic acid
solution was used as a stock solution. To estimate the reproduc-
ibility of the presented nanocomposite materials, three different
sensors were evaluated for each nanocomposite composition.
High-Resolution Transmission Electron Microscopy (HReTEM),
using a JEM-1400 unit with an acceleration voltage of 120 kV, was
used in order to characterize the morphology of the different car-
bon materials (raw graphite, GO and rGO). ~ 0.1 mg of sample was
dispersed in 10 mL of Milli-Q water as solvent and then placed in
ultrasound bath for 2 h. Finally, a drop of this solution was placed
on a grid and left to dry before HR-TEM analysis. Measurements of
nanocomposites topography were made by Confocal Microscopy
3D (CM3D). A Leica DCM 3D unit to 150 magnifications was used
for measuring fifty-seven profiles in three different areas for the
different nanocomposite electrodes (ISO 4287). Electrical resistance
measurements were performed by means of a digital multimeter
(Fluke, Everett, WA, USA). In order to estimate the reproducibility of
the hand-made fabrication three equal nanocomposite electrodes
were fabricated and evaluated for each composition (from 8% to
20% in conducting filler nanomaterial, w/w). In addition, five
different points were tested for each composition of three different
nanocomposite electrodes (n ¼ 15) in order to study the repeat-
ability of the measurements. Measurements of topography and
current were obtained simultaneously by means of a Current
Sensing Atomic Force Microscopy (CSAFM), (PicoSPM, Molecular
Imaging, USA). The surface measurements were done using the
current sensing mode: CSAFM and rectangular diamond coated tips
(Nanoworld, Switzerland) with an estimated constant spring of
72 N m1. The optical characteristic of the graphene-based mate-
rials were monitored using UVevis spectroscopy (Agilent Tech-
nologies, Cary 60 UveVis scanning spectrometer). Raman spectra
were recorded with a Renishaw 1000 micro e Raman system fitted
with a Leica microscope and Grams Research™ analysis software.
The excitation source used was a 633 nm HeNe laser (Renishaw
RL633). The 50 objective lens was capable of focusing the beam to
a spot size ~ 2e3 mm in diameter.
2.3. Synthesis of GO and rGO
Graphene Oxide (GO) was synthesized from natural flaked
graphite via Hummers method [16]. The product was washed
several times through centrifugation (4000 rpm) with a 10%
aqueous solution of hydrochloric acid followed by copious amounts
of water until the pH was 6. Finally, the product was dried over-
night at 80
C.
Then, rGO was obtained from the reduction of GO with ascorbic
acid, following the literature procedure [17]. Afterwards, the
product was centrifuged at 4000 rpm for 10 min and washed
several times with Milli-Q water until the pH reached ~6. The
resultant product was dried overnight at 80
C.
2.4. Nanocomposite electrode preparation
Handmade working nanocomposite electrodes were prepared
as follows: polymer Epotek H77 and its corresponding hardener
 ~ oz et al. / Composites Science and Technology 125 (2016) 71e79
J. Mun 73

compound were mixed in the ratio of 20:3 (w/w) before loading

different amounts of rGO, from 8% to 20% (w/w). The nano-
composites were then manually homogenized for 30 min. For the
electrode construction, the nanocomposite was placed into a cy-
lindrical PVC tube (i.e., 6 mm and i.d., 20 mm length) containing a
copper disk soldered to an electrical connector end. The final paste-
filled cavity was 3 mm long inside the PVC tube [18]. Then, the
nanocomposite paste electrodes were allowed to harden at 80
C
for 24 h. Afterwards, the electrode surfaces were polished with
different sandpapers of decreasing grain size (800 and 1200) and
finally with alumina paper (polishing strips 948201, Orion) in order
to obtain a reproducible electrochemical surface. The resultant
geometric area for the final electrodes was 28 mm2. It is important
to highlight that three equal electrodes were fabricated and eval-
uated for each graphene composition in order to estimate the
reproducibility of the developed nanocomposite electrodes.

3. Results and discussion

3.1. Characterization of GO and rGO

High-Resolution Transmission electron microscopy (HR-TEM)

was employed to monitor the extent of exfoliation of GO after
dispersion in H2O using ultrasonication. TEM images from Fig. 1
show clearly the effect of oxidation on the dispersibility of the
material in water. Fig. 1A shows bulk graphite after deposition onto
a lacy carbon TEM grid. Fig. 1B depicts a representative image of the
GO after oxidation, suspension and deposition from H2O. Finally,
Fig. 1C shows a representative image of the rGO, which was ob-
tained after GO reduction with ascorbic acid.
UVevis spectra recorded for GO and rGO can be seen in Fig. 2A.
The main spectroscopic features associated with optical absorption
in GO are the due to electronic transitions within eC]C bonds
(231 nm) and eC]O bonds (~300 nm). After reduction has taken
place, the band associated with eC]O functionality is removed
from the spectra while the band located at 231 nm due to in-plane
eC]C bonds shifts to the red (267 nm). This shift is known to be
associated with the increased in-plane hybridisation of the rGO [19]
Fig. 2. A) UVevis spectra recorded for aqueous suspensions of a) GO and b) rGO. B)
and agrees well with the findings on the bibliography [17]. Raman spectra recorded for a) graphite, b) GO and c) rGO deposited on Si/SiO2
Raman spectroscopy is an extremely useful technique for anal- (300 nm) substrates.
ysis of carbon based materials. Fig. 2B shows the Raman spectra of
the three studied samples, which are composed of 2 peaks: the
disorder band (D band) located at ~1330 cm1 and the graphitic associated with the isolation of carbonecarbon double bonds in the
band (G band) located at ~1580 cm1 [20]. A large increase in the lattice, which causes resonance at a higher frequency [21].
intensity of the D band can be observed upon oxidation of the According with these results, it can be concluded that GO was
graphite to the graphene oxide, which is associated with increased successfully synthesized from bulk graphite and then efficiently
level of disorder in the material. This oxidation causes the G band of reduced with ascorbic acid for obtaining the rGO nanofiller
GO band to red shift (1578e1582 cm1). This shift has been material.

Fig. 1. HR-TEM images recorded for A) graphite, B) GO and C) rGO deposited onto Lacy carbon grids from aqueous suspension.
74 ~oz et al. / Composites Science and Technology 125 (2016) 71e79
J. Mun

3.2. Electrical characterization of rGO/epoxy nanocomposite

materials

The electrical properties of rGO/epoxy nanocomposite elec-

trodes were investigated applying the well-known percolation
theory [22]. The percolation curve was constructed by varying the
rGO loading in the epoxy matrix from 8% to 20% (w/w). While
nanocomposites containing less than 8% of rGO presented insu-
lating properties with resistivity values ~ ∞, those containing more
than 20% of rGO gave poor mechanical stability. From an analytical
point of view, the rGO loading has to be high enough to guarantee a
conducting pathway and sufficiently low bulk resistivity due to a
lower resistivity value which produces a higher sensitivity [18].
After the experiments, three different conductive regions were
determined regarding the electrical behaviour (see Fig. 3): High
resistance zone (HRC), near-percolation zone or second percolation
composite zone (NPC) and low resistance zone (LRC). According to
the results, the variation of the resistivity was negligible between
13% and 20% of rGO loading, demonstrating that the nano-
composites lying between such composition intervals are suitable
for electroanalytical measurements.
Finally, Fig. 3 also shows a significant decrease in the standard
deviation of the resistivity values when the rGO/epoxy composition
ratio reached the NPC zone. Taking these observations into account,
it can be concluded that nanocomposite lying between the NPC
zone and the LRC zone are suitable for electroanalytical measure-
ments, resulting in more reproducible nanocomposite electrodes.

3.3. Voltammetric characterization of rGO/epoxy nanocomposite

electrodes

CV was applied in order to electrochemically characterize the

different rGO/epoxy nanocomposite electrodes. Ten different rGO/
epoxy nanocomposite electrodes composition (from 8% to 20%)
were fabricated. Firstly, the optimum scan rate was study from 10 to
100 mV s1, finding the optimum scan rate at 50 mV s1. Secondly,
different parameters such as peak height (Ip), peak separation po-
tential (DE) and electroactive area (A) were extracted from the Fig. 4. Electrochemical experiments recorded in a 0.01 M [Fe(CN)6]3/4 solution for
cyclic voltammograms (see Fig. 4A), which are summarized in the different rGO/epoxy nanocomposite compositions. A) Cyclic voltammograms (scan
Table 1. The kinetic processes of those species are strongly depen- rate: 50 mV s1) and B) Electrochemical impedance spectra. The insert shows the
equivalent Randles circuit used for fitting.
dent on the state of the electrode surface. The electroactive area (A)

can be calculated in terms of peak current (Ip, A) and scan rate (y,
V s1) according to the modified Randles-Sevcik equation, Equation
(1) [23]:

*
Ip ¼ 3:01  105 $n3=2 ðaDred yÞ1=2 A$Cred (1)

This equation is adequate for electron transfer-controlled pro-

cesses [24], where n is number of electrons (n ¼ 1), Cred* is the bulk

concentration of the electroactive species (Cred * ¼ 0.01 M), D

Fig. 3. Percolation curve obtained for the rigid conducting nanocomposite electrodes
based on rGO/epoxy, where the rGO load varies between 8% and 20%. The trend follows
the percolation theory, showing three different conductive regions: HRC, NPC and LRC.
Experimental errors was estimated as a standard deviation (n ¼ 15).
red
corresponds to the diffusion coefficient of the reduced species
(Dred ¼ 6.32  106 cm2 s1), a represents the symmetry factor
which was taken approximately as 0.5 and y represents the scan
rate (y ¼ 0.05 V s1). According to voltammetric results (see
Table 1), peak height increased with the rGO loading, due to an
increase in the electroactive area. Simultaneously, a decrease of
peak separation was also observed, related to an enhancement of
the electron transfer rate.
In addition, the exchange current (i0) from Tafel plots (log cur-
rent density vs. potential) has also been evaluated. i0 is a parameter
which provides information about the reversibility of the electron
transfer process. From this value it is possible to evaluate the charge
transfer resistance (Rct) through the Equation (2):
 ~ oz et al. / Composites Science and Technology 125 (2016) 71e79
J. Mun 75

Table 1
Electrochemical parameters for the different nanocomposite compositions were recorded in a 0.1 M KCl containing 0.01 M [Fe(CN)6]3/4. i0 corresponds to the exchange
current, Rct to the charge transfer resistance, Ip to peak current, A to electroactive area and DE to the peak separation potential. Rct$A and REIS
ct $A corresponds to the Rct obtained
by voltammetric and EIS measurements, respectively, and normalized with respect to the electroactive area.

% rGO i0 (mA) Rct (kU) Ip (mA) A (cm2) DE (V) Rct$A (U$cm2) ct $A (U$cm )
REIS 2

8 18.7 1.4 250 0.21 1.57 283 134

10 21.3 1.2 260 0.22 1.44 259 95
11 38.0 0.67 330 0.28 1.02 184 94
12 61.6 0.41 419 0.35 0.93 144 120
13 155 0.16 511 0.43 0.41 70 121
14 189 0.13 580 0.48 0.34 65 90
15 236 0.11 669 0.56 0.31 60 93
16 273 0.093 699 0.58 0.31 54 54
18 303 0.084 717 0.60 0.31 50 27
20 309 0.082 788 0.66 0.35 54 26

RT
i0 ¼
nFRct
The trend observed is a decrease of this parameter with
decreased loadings of rGO (see Table 1); this observation indicates
the strong relationship between electrochemical reactivity and the
surface characteristics of the conducting material. As the carbon
load increases, the probability of having more electroactive sites
increases and hence also the electrode kinetics. Accordingly, a low
value in the charge transfer resistance allows for increased sensi-
tivity and improved electroanalytical capabilities of the electrode.
By normalizing charge transfer resistance with respect to the
electroactive area (Rct$A), it is possible to observe that this
parameter also decreased with the increase of A. The parameter
Rct$A should be constant for conventional metal electrode in-
terfaces. However, in the case of carbon nanomaterials, this is not
expected as they are known to exhibit electrochemical anisotropy
[25].
3.4. Impedimetric characterization of rGO/epoxy nanocomposite
electrodes
EIS technique has been carried out in order to obtain resistance/
impedance information about the different nanocomposite electrode
compositions. This technique uses a model circuit to describe indi-
vidual circuit elements which are based on the physical electro-
chemistry system, such as solution resistance (RU), which is
dependent on the ionic concentration, the type of ions and also the
electrode area; charge transfer resistance (Rct), which is inversely
proportional to the electron transfer rate; double-layer capacitance
(Cdl), which is directly related to the charging or background current
and Warburg impedance (Zw), which arises from mass-transfer lim-
itations and can be used to measure effective diffusion coefficients. In
this way, these physical parameters were successfully evaluated for
each nanocomposite electrode composition, which were obtained by
fitting the impedance spectra to a Randles circuit (Fig. 4B, inset). Using
such an approach, it is possible to obtain a general trend of these
physical parameters as a function of rGO loading.
Fig. 4B shows the Nyquist plots obtained for different rGO/epoxy
electrode compositions. The Nyquist plots of the EIS consist of a
semicircular portion and a linear portion, which corresponds to the
electron transfer limited process and the diffusion limited process,
respectively [26]. As is shown in Fig. 4B, the impedance behaviour is
mainly dominated by a semicircle, representing kinetic-controlled
electrode process. Thus, nanocomposite electrodes containing low
rGO loadings (from the HRC zone to the NPC zone) appeared to be
dominated by a big diameter semicircle, demonstrating that only
the kinetic-controlled electrode process were present in the
recorded frequency range. However, for nanocomposites
containing high rGO loading (from the NPC zone to the LRC zone)
the impedance behaviour was dominated by a small diameter
(2)
semicircle and the diffusion-controlled process started to be dis-
cerned in some cases at low frequencies.
The parameter RU is the one that has a direct relation with the
dry resistance taken for the percolation plot of Fig. 3. Fig. 5A shows
the variation of the ohmic resistance as a function of the rGO
composition and it can be seen that the RU values decreased with
increased rGO loading. This is due the fact that at low carbon loads
(<14% of rGO loading), the ohmic resistance is dominated by the
nanocomposite resistance whereas at higher carbon loads (14% of
rGO loading), the ohmic resistance decreases to low values and it is
more dominated by the solution resistance. According to these
results, this behavior is in accordance with those obtained by
percolation curve.
Concerning Rct, this parameter is well known as a useful value
which reflects the ability of an electrode reaction, which can be
measured by EIS from the semicircle diameter in the Nyquist plot.
As is shown in Fig. 5B, nanocomposites between the NPC zone and
the LRC zone presented lower Rct values, demonstrating that the
nanocomposites with low charge transfer resistances are the most
appropriate to be used in electrochemical measurements.
Normalizing the EIS Rct values with respect to the A (REIS ct $A), it is
possible to observe that there is also a decrease in the value when
the electroactive area increases, following the observed tendency in
CV performance (see Table 1).
Impedimetrically, Cdl is also an important parameter which is
directly related to the charging or background current and
inversely proportional to the signal-to-noise ratio. The electrode
capacitance can be determined nearly exclusively by the exposed
carbon on the electrode surface, as is reflected in Fig. 5C. An in-
crease of the Cdl value with the increase loading of conducting
nanofiller material can be observed. This fact is due to only a
fraction of conductive area is exposed to the solution and the other
fraction is occupied by the insulating polymer. Thus, a high pro-
portion of rGO loading increases the conducting area, the surface
roughness and porosity of the nanocomposites [27].
From an analytical point of view, an electrode must have a rapid
response time, low limit of detection and high sensitivity. According
to the impedance results and taking this into consideration, the in-
terval between 12% and 14% of rGO loading is considered the opti-
mum to obtain the best electroanalytical response due to small
variations in compositions around this optimal range do not produce
high changes in the electrochemical nanocomposite behaviour.
3.5. Electroanalytical response of rGO/epoxy nanocomposite
sensors
Ascorbic acid was used as a model analyte for evaluating the
electroanalytical characteristics of the rGO/epoxy nanocomposite
76 ~oz et al. / Composites Science and Technology 125 (2016) 71e79
J. Mun
Fig. 5. EIS values of A) RU, B) Rct and C) Cdl, with their corresponding standard devi-
ation for the different rGO loading electrodes, using the redox probe [Fe(CN)6]3/4.
sensors in terms of hydrodynamic amperometric measurements
(see Fig. 6). It is important to highlight that three different nano-
composite sensors were prepared and evaluated in order to
compare their sensitivity. The measurements, which were carried
out in triplicate (n ¼ 3), yielded a relative standard deviation (RSD)
of 5.8% and 0.4% for the NPC and LRC sensors, respectively. These
results confirm that the preparation method is highly reproducible.
Amperometric results which are summarized in Table 2
demonstrated that when the rGO loading increases from 13% to
20%, both Limit of Detection (LOD) and Limit of Quantification
(LOQ) showed a remarkable increase; although the most concen-
trated nanocomposites prepared (20% in rGO loading) contain a
higher number of conducting particles. These results can be
explained since a lower rGO load becomes in an increase of the
signal-to-noise ratio (Cdl), resulting in an 83% and 72% improve-
ment in LOD and LOQ, respectively. Accordingly, these results
suggest that the rGO loading of 13% is the optimal composition of
nanofiller material in the nanocomposite electrode, since it can
achieve a good sensitivity but more importantly, lower limits of
detection.
Comparing these results with those obtained previously in our
research group for optimum graphite/epoxy composite sensors
(15% w/w in graphite loading, NPC sensor), better LODs have been
obtained with the optimized graphene-based nanocomposite
sensors (13% w/w in rGO loading, NPC sensor). Thus, while the
optimized sensors based on graphite presented a LOD of 17 mg L1
[28] for ascorbic acid, this parameter was significantly enhanced for
the optimized graphene-based sensor (3 mg L1). This fact resulted
in an improvement of 0.8 decades of concentration regarding to
graphite/epoxy NPC sensors for the model analyte. Accordingly, the
simple bulk graphite modification for synthesizing the rGO nano-
material presents important benefits from an analytical point of
view, resulting enhanced nanocomposite sensors because of they
provide lowest LODs. Otherwise, comparing this results with the
ones obtained in our research group for optimized carbon nano-
tubes/epoxy nanocomposite sensors (10% w/w in carbon nanotubes
loading, NPC sensor), an enhancement in the LOD of 0.4 decades of
concentration was also obtained [29]. In comparison with carbon
nanotubes, graphene presents significant advantages such as it
does not contain metallic impurities and is produced from flacked
graphite, which is economical and accessible.
3.6. Morphological characterization of rGO/epoxy nanocomposite
electrodes
CSAFM, CM3D and SEM techniques were carried out in order to
gain an insight of the surface characteristics of these two types of
nanocomposites (NPC and LRC electrodes).
Firstly, the CSAFM technique was used in order to obtain qual-
itative information about the size, shape and distribution of the
conducting nanomaterial in the nanocomposite structures. Fig. 7(A
to B) presents the most significant images obtained during the
surface study of the NPC and LRC electrodes. CSAFM images allow
the observation that even though both materials have a macro-
electrode appearance, the NPC surface includes many separated
conductive microzones on its surface where the electronic transfer
is produced, similar to a microelectrode array behaviour. This fact
offers the electrode the possibility to generate radial diffusion
where the electroactive species diffuse on its surface by the edge
effect [30]. However, for the LRC electrodes only the linear diffusion
is contemplated. Thereby, such radial diffusion, provides an in-
crease of the total mass transfer [31], which favours positively the
electroanalytical signal because Cdl decreases (see Fig. 5C) and
consequently the limit of detection improves (see Table 2). The
literature shows works where the amperometric behaviour of
composite-based electrodes compared to microelectrode arrays is
studied [32]. In these cases, it is important to optimize the distance
between microelectrodes (conductive microzones) in order to
obtain an optimal signal-to-noise ratio. Due to the random struc-
ture of the presented nanocomposites, the distance between
conductive microzones are not easy to control. Nevertheless, this
 ~ oz et al. / Composites Science and Technology 125 (2016) 71e79
J. Mun 77

Fig. 6. Calibration curve of current vs. concentration of ascorbic acid for A) NPC and B) LRC sensors. The calibration curves are represented with their corresponding error bars
(n ¼ 3). Dynamic responses of C) NPC and D) LRC sensors were carried out in a 0.01 M KNO3/HNO3 background electrolyte solution.

Table 2
Calibration parameters for NPC and LRC electrodes were obtained by hydrodynamic amperometry, using ascorbic acid as the model analyte in a 0.01 M KNO3/HNO3 background
electrolyte solution; working potential: 600 mV. LOD and LOQ were calculated three times (n ¼ 3) and they are presented with their respectively 95% confidence interval.

Sensor rGO loading (%) LOD (mg$L1) LOQ (mg$L-1) Sensitivity (mA$L$mg1) r2 (n ¼ 13)

NPC 13% 3±2 7±5 0.61 ± 0.05 0.999

LRC 20% 18 ± 8 25 ± 8 0.284 ± 0.002 0.999

morphological characterization makes it possible to slightly control
the distance between the conductive microzones by a decrease or
an increase of the rGO loading.
In addition to CSAFM experiments, surface roughness of both
rGO/epoxy nanocomposite electrodes was also compared by CM3D.
Fig. 7(C to D) presents significant images obtained during the
electrode surface study. When the electrode roughness increases,
the probability of more conductive material exposed on the elec-
trode surface also increases and consequently, its electroactive area.
According to confocal results, which are summarized in Table 3, LRC
electrodes presented a roughness value 34% higher than the
optimized-electrodes (NPC electrodes). These results are consistent
with the A values obtained by CV (see Table 1), where an increment
of 35% in A was also achieved for the LRC electrodes.
Finally, SEM images allowed us to observe qualitatively the
roughness and the porosity of the nanocomposite surface. Fig. 7(E
and F) presents the comparison of the SEM images for the two
different rGO loading studied (the NPC and the LRC electrodes). The
surface of LRC nanocomposite electrode (20% in rGO w/w) was
rougher than in the case of the NPC nanocomposite electrode (13%
in rGO w/w). In addition, it is also possible to observe how the
morphological structure of the surface is less homogeneous and
more porosity when the amount of conducting nanofiller increases,
in agreement with the results obtained by CM3D technique.
Morphological results demonstrated that through optimizing
the graphene/polymer electrode composition it is possible to
improve the electroanalytical features (lower limits of detection
and quantification), resulting in more efficient sensors.
4. Conclusions
For first time, a powerful electrochemical strategy has been
applied to develop efficient graphene-based electrochemical
nanocomposite sensors with an optimized analytical response in
terms of high signal-to-noise ratio, rapid response and low limit of
detection. Electrochemical techniques, such as EIS and CV allowed
us to optimize the nanocomposite composition in terms of (i)
ohmic resistance, which is related to the percolation resistivity; (ii)
78 ~oz et al. / Composites Science and Technology 125 (2016) 71e79
J. Mun

Fig. 7. Conductance mapping obtained by CSAFM for A) NPC and B) LRC and their corresponding topographic images obtained by CM3D (C and D) and SEM (E and F).

Table 3 that these parameters are relevant for electroanalytical purposes,

Roughness values collected from surface analysis. Data are the mean and standard an accurate electrochemical characterization results mandatory in
deviation (SD) from 57 profiles of three areas examined in each series (ISO 4287).
order to define the optimal graphene/polymer composition ratio.
Electrodes rGO loading (%) Raa (mm) Rqb (mm) Rzc (mm) Electrical and electrochemical experiments determined that the
Mean SD Mean SD Mean SD optimum rGO/epoxy nanocomposite composition (the NPC) is be-
tween a 12% and 14% of rGO loading. Small variations in composi-
NPC 13 0.50 0.11 0.63 0.14 2.57 0.55
LRC 20 0.76 0.12 0.93 0.14 3.71 0.63 tion around this optimal range do not produce high changes in the
a
electrochemical response behavior, resulting in more reproducible
Aritmetical mean deviation of the assessed profile.
b
The root mean square deviation of the assessed profile.
sensors. The feasibility of this approach in terms of electroanalytical
c
Maximum height of the profile. detection was confirmed using ascorbic acid, obtaining better
sensitivity, LODs and LOQs than those non-optimized. In addition,
morphological experiments verified that the electrochemical and
charge transfer resistance, which is related with the heterogeneous electroanalytical response of the graphene-polymer nano-
electron transfer rate and also depends on the surface electro- composite sensors closely depends on their composition ratio.
chemical reactivity and (iii) double-layer capacitance which can be Comparing the graphene-based sensors results with those ob-
correlated to the background current, an important parameter to tained from the bulk graphite-based sensors, it can be concluded
minimize in order to enhance the signal-to-noise ratio. Considering that the simple exfoliation of graphite significantly enhanced the
 ~ oz et al. / Composites Science and Technology 125 (2016) 71e79
J. Mun
demanded requirements for electrochemical sensors, improving
82% the LOD for the studied model analyte.
Finally, we believe that this study opens up routes for using the
present characterization protocol to optimize graphene-polymer
electrochemical nanocomposite (bio)sensors for improving their
electroanalytical performance. Thereby, this work shows a novel
perspective in the characterization and optimization of graphene-
based nanocomposite materials which will allow for extension of
this approach for various polymeric matrices as well as other 2D
nanomaterials related to the graphene family.
Acknowledgement
This work was supported by the project CTQ2012-36165, the
Science Foundation Ireland (SFI 12/IA/1300 and Amber project) and
the Ministry of Education and Science of the Russian Federation
~ oz thanks Universitat Auto
(Grant no. 14.B25.31.0002). J. Mun  noma
de Barcelona (UAB) for the award of PIF studentship.
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