Special Issue ADVANCED MATERIALS
Full Paper
DOI: 10.1002/elan.201500168
Screen-Printed Electrodes Modified with Carbon
Nanomaterials: A Comparison among Carbon Black,
Carbon Nanotubes and Graphene
Stefano Cinti,[a, b] Fabiana Arduini,*[a, b] Marilena Carbone,[a, b] Lucia Sansone,[c] Ilaria Cacciotti,[d, e]
Danila Moscone,[a, b] and Giuseppe Palleschi[a, b]
Abstract: In this work a comparative study using Screen-
Printed Electrodes (SPEs) modified by drop casting with
Carbon Black, Single Walled Carbon Nanotubes¢COOH,
Graphene Oxide, and reduced Graphene Oxide is report-
ed. The carbon nanomaterials employed were character-
ized by X-ray photoelectron and Raman spectroscopy,
while the modified SPEs have been morphologically and
Keywords: Screen-printed electrode · Carbon black · Graphene · Carbon nanotubes · Drop casting
1 Introduction
Carbon is present in several allotropic forms ranging
from graphite to diamond, through the most recently dis-
covered fullerene, nanotubes and graphene. Carbon, in all
its forms, finds application in many fields: catalysis [1],
batteries [2], solar cells [3], water purification [4], energy
storage [5], automotive [6], metallurgy [7]. In particular,
carbon allotropes are most used in matters of chemical
sensors, especially in the electrochemical field. Sensors
based on these carbonaceous materials have been report-
ed in literature with applications in many fields such as
food [8], clinical [9], environmental [10], in combination
with biomolecules such as enzymes [11], aptamers [12],
as well as with metallic nanoparticles [13], macrocyclic
compounds [14], polymers [15]. The first carbon-based
electrodes are those made of graphite. They are charac-
terized by a wide operating potential range due to their
chemical inertness. A plethora of works followed on the
use of many types of electrodes based on carbon (e.g.
glassy carbon, carbon paste) [16]. The discovery of fuller-
ene, carbon nanotubes (CNTs) and graphene has impact-
ed very positively in the electrochemical field. The
former did not and still does not represent a big deal
toward the development of chemical sensors. The latter
ones hold a leading role in the current sensor scenario
[17, 18], thanks to their unique properties, e.g. electron
mobility, low resistivity. The presence of CNTs or gra-
phene on the surface of the working electrodes can im-
prove the analytical performances by enhancing the elec-
tron transfer between the surface of modified electrodes.
Because of the advantages of these nanomaterials, various
procedures have been developed to produce them [19–
23], although sometimes not in large quantities, or, as far
as graphene is concerned, not in a single sheet (pristine).
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electrochemically characterized. Nanoengineered SPEs
have been tested with ferricyanide, NADH, ascorbic acid
and cysteine. We observed valuable electroanalytical per-
formances of Carbon Black with the advantage to be i)
cost-effective ii) suitable to obtain stable and homoge-
nous dispersion and iii) mass-producible following a well
established route.
In recent years another interesting carbonaceous, though
“less noble” nanomaterial, is becoming utterly interesting
due to its excellent conductive and electrocatalytic prop-
erties: Carbon Black (CB). CB finds applications as sensi-
tive and filler material, and it is cost-effective (about
1 euro/kg). The advantages to use CB for analyte detec-
tion were demonstrated by our research group [24]. Af-
terwards, several investigations carried out both by us
and by other groups confirmed the excellent electrocata-
lytic properties of CB [25–31].
The necessity in the analytical field is always to develop
methods and platforms capable to be very sensitive, af-
fordable, and easy to use. A valuable alternative is repre-
sented by the cost-effective printed electrodes which can
be easily miniaturized and obtained with hand-made in-
[a] S. Cinti, F. Arduini, M. Carbone, D. Moscone, G. Palleschi
Dipartimento di Scienze e Tecnologie Chimiche, Universit—
di Roma Tor Vergata
Via della Ricerca Scientifica, 00133 Rome, Italy
*e-mail: fabiana.arduini@uniroma2.it
[b] S. Cinti, F. Arduini, M. Carbone, D. Moscone, G. Palleschi
Consorzio Interuniversitario Biostrutture e Biosistemi
“INBB”
Viale Medaglie dÏOro 305, 00136 Rome, Italy
[c] L. Sansone
Istituto per Polimeri, Compositi e Biomateriali
CNR p.zzale E. Fermi I, 80055 Portici (NA) Napoli, Italia
[d] I. Cacciotti
Universit— degli Studi di Roma “Niccol! Cusano”, Via Don
Carlo Gnocchi 3, 00166 Rome, Italy
[e] I. Cacciotti
Consorzio Interuniversitario Nazionale per la Scienza
e Tecnologia dei Materiali (INSTM)
Supporting Information for this article is available on the
WWW under http://dx.doi.org/10.1002/elan.201500168.
Electroanalysis 2015, 27, 2230 – 2238 2230

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strumentation upon necessity. Carbon ink is a suitable
platform to be modified with various kinds of material
e.g. nanomaterials, organic macromolecules or polymers,
due to its the resistance to a wide range of solvents, low
background current and broad potential windows of ap-
plication. However, the (nano)modified surface leads to
a specific electrocatalytic behaviour depending solely on
the nature of the modifier. This paper reports a morpho-
logical and electrochemical characterization of screen-
printed electrodes (SPEs) nanoengineered by means of
SWCNTs, graphene and CB. The different modified elec-
trodes will be characterized by scanning electron micros-
copy (SEM), electrochemical impedance spectroscopy
(EIS) and cyclic voltammetry (CV). Furthermore, all the
modified SPEs have been tested in the presence of sever-
al relevant molecules such as nicotinamide adenine dinu-
cleotide reduced (NADH), ascorbic acid (AA) and cys-
teine, obtaining important information related to their an-
alytical performance.
2 Experimental
2.1 Apparatus
Voltammetric and amperometric measurements were car-
ried out using a portable potentiostat PalmSens (Palm In-
struments, The Netherlands). The morphology of the pre-
pared SPEs was analyzed by observation at field emission
scanning electron microscopy (FEG-SEM, Leo Supra 35,
UK). Raman spectra were acquired by a Renishaw in Via
Reflex Raman spectrometer (United Kingdom) using
514.5 nm excitation source. XPS measurements were per-
formed by using an ESCALAB MkII (UK) (VG Scientif-
ic) spectrometer (Thermo Scientific, West Sussex, UK),
equipped with a hemispherical analyzer. Mg Ka photons
(hn = 1253.6 eV) were used to induce electron emission.
The base pressure was lower than 2 × 10¢10 mbar.
Electrochemical impedance spectroscopy (EIS) meas-
urements were carried out in the same cell with a PC-
controlled Autolab. A sinusoidal voltage perturbation of
10 mV amplitude was applied over the frequency range
10 kHz to 0.01 Hz with 10 measurement points per fre-
quency decade. For the fitting of the data obtained by
EIS, Z-views software (Scribner Associates, Inc.) was
used.
2.2 Reagents
Commercial CBN220 of industrial standard grade was ob-
tained from Cabot Corporation (Ravenna, Italy),
SWCNTs were purchased from Nanoshel (average diame-
ter 1–2 nm, average length 1.5 mm). Graphene oxide
powder was purchased from Cheap Tubes, Inc. (USA).
All reagents used were obtained from Sigma (St. Louis,
MO).
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Special Issue ADVANCED MATERIALS
2.3 SPE Preparation
SPEs were produced with a 245 DEK (Weymouth, UK)
screen-printing machine. Graphite based ink (Electrodag
423 SS) from Acheson (Milan, Italy) was used to print
the working and counter electrode. Silver ink (Electrodag
477 SS) was used to print the reference electrode. The
substrate was a flexible polyester film (Autostat HT5) ob-
tained from Autotype Italia (Milan, Italy). The diameter
of the working electrode was 0.3 cm resulting in a geomet-
ric area of 0.07 cm2.
2.4 Carbon Nanomaterial Dispersion
2.4.1 Carbon Black Dispersion
The dispersion of CB was prepared by adding 10 mg of
CB powder to 10 mL of solvent (a mixture dimethylfor-
mamide (DMF) : water (1 : 1)) sonicated for 60 min at
59 kHz.
2.4.2 Single Walled Carbon Nanotubes Dispersion
The dispersion of Single Walled Carbon Nanotubes –
COOH (SWCNTs-O) was prepared by adding 10 mg of
SWCNTs powder to 10 mL of solvent (a mixture dime-
thylformamide (DMF):water (1 : 1)) sonicated for 60 min
at 59 kHz.
2.4.3 Graphene Oxide and Reduced Graphene Oxide
Dispersions
The graphene oxide, purchased from Cheap Tubes, was
synthesized from purified natural graphite by the modi-
fied Hummers method. Ten milligrams of GO powder
was suspended in 5 mL of distilled water and sonicated
by means of an ultrasonic processor (Misonix Incorporat-
ed Ultrasonic liquid Processors, USA) for 4 hours at
room temperature so as to produce a stable yellow-brown
colloidal suspension of GO sheets. Afterwards, 10 mg of
GO powder were added to 10 mL of solvent (a mixture
dimethylformamide (DMF):water (1 : 1)) sonicated for
30 min at 59 kHz.
The reduced graphene oxide (rGO) was synthesized as
follows: GO (100 mg) was loaded in a 250-mL roundbot-
tom flask and water (100 mL) was then added, yielding
an inhomogeneous yellow-brown dispersion. This disper-
sion was sonicated using a ultrasonic bath until it became
clear with no visible particulate matter. Hydrazine hy-
drate (1.00 mL, 32.1 mmol) was then added and the solu-
tion heated in an oil bath at 100 8C under a water-cooled
condenser for 24 h over which the reduced GO gradually
precipitated out as a black solid. This product was isolat-
ed by filtration, copiously washed with water and metha-
nol, and dried. Afterwards, 10 mg of rGO powder were
added to 10 mL of solvent (a mixture dimethylformamide
(DMF) : water (1 : 1)) and sonicated for 60 min at 59 kHz.
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2.5 Modification of Screen-Printed Electrodes
The SPEs are modified with the different dispersions
(CB, SWCNTs-O, GO, rGO) via drop casting approach:
a small volume (6 mL) of the dispersion was casted onto
the working electrode surface of the SPE in three steps of
2 mL. Modified SPEs are reported in the text as CB-SPE,
SWCNTs-O-SPE, GO-SPE and rGO-SPE.
2.6 Amperometric Measurements
Measurements of the different analytes (ascorbic acid,
cysteine and nicotinamide adenine dinucleotide reduced)
were performed with each different modified-SPEs using
amperometric batch analysis in 10 mL of stirred 0.05 M
phosphate buffer solution + 0.1 M KCl, pH 7.0 with a se-
lected applied potential. When a stable baseline current
was reached, the analyte was added and the response was
recorded.
3 Results and Discussions
3.1 Physical Characterization
The first study was devoted to perform the structural
characterization of the carbon nanomaterials. CB, Gra-
phene Oxide (GO), reduced Graphene Oxide (rGO), and
Single Walled Carbon Nanotubes ¢COOH (SWCNTs-O),
were characterized by means of XPS.
The XPS spectra were taken in several binding energy
ranges to identify the elemental species present on the
different compounds and their oxidation states. The most
significant spectra, i.e. the ones in the range including the
C1s threshold, are reported in Figure 1. The CB spectrum
in the C1s range displays a single peak at 284.3 eV, with
a shake-up peak at 290.3 eV typical of graphite and
graphite-like structures [32]. Traces of oxygen are present
in the order of 2 % atomic weight.
GO is strongly oxidized. At least three components can
be identified by C1s line-shape analysis, at 284.8 eV,
286.8 eV and 288.6 eV. These values are attributed to the
extended sp2 domains (lower binding energy peak), C=O
and O¢C=O, (higher energy peaks at + 2.0 and + 3.8 eV,
respectively) [33]. The ratio between the three compo-
nents is 7 : 7 : 1. The total oxygen amount is 34 in atomic
%, and the corresponding O1s spectrum is characterized
by two components in ratio compatible with the C1s oxi-
dized ones.
The reduction of the GO, drastically reduced the
oxygen content, from 34 % to 12 %, though leaving traces
of nitrogen. Compared to the oxidized form, the overall
number of C1s components increases, due to the presence
of sp2-hybridized related peak at 284.3 eV and associated
shake-up peak at 290.3 eV. The intensity of the C1s peaks
related to C=O and O¢C=O moieties is significantly re-
duced and a new peak appears at 285.9 eV, which can be
attributed to partially reduced ¢C¢NOH species [33].
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Special Issue ADVANCED MATERIALS
Fig. 1. XPS spectra of CB, GO, rGO, SWCNTs-O, (black lines)
and corresponding line shape analysis (the green lines are the
different components, the blue line is the total envelop).
C1s spectrum of the SWCNTs-O is characterized by
the presence of sp2-related component and associated
shake-up peak (284.3 eV, 290.3 eV). Furthermore, three
peaks are associated to the intermediate and highly oxi-
dized states (at 285.3 eV, 286.4 eV, 288.6 eV). The sp2 -re-
lated and the oxidized components are in a ratio
5 : 3 : 2 : 2. The total amount of oxygen is 22 atomic %.
Next, the Raman spectra in the 1000–2000 cm¢1 range
of the GO, rGO, SWCNT-O and CB are reported in
Figure 2. The Raman spectrum of GO (Figure 2 black
solid line) displays a D-band at 1340 cm¢1 and a broad G-
band at 1580 cm¢1. The D peak can be related to the
biding to oxygen species. The G peak duly corresponds to
the first order scattering of the E2G phonon mode of sp2
C atoms [34].
The Raman spectrum of the rGO (Figure 2 red solid
line, superposed to the GO line) also displays G and D
bands at 1580 cm¢1 and 1340 cm¢1, respectively. Com-
pared to GO, the D peak intensity of the rGO increases.
In particular, the ID/IG ratio (between the peak areas) in-
creases from 1.19 to 1.26. This may indicate the formation
of more sp2 domains of smaller dimension, which is
common in graphene oxide reduction procedures [35, 36].
In the SWCNT-O, the D band is peaked at 1363 cm¢1 and
the G band at 1593 cm¢1. The ID/IG ratio = 1.23. Further-
more, the D band is broad and asymmetric. Shape and in-
tensity of D-mode correspond to the sp3-hybridized
carbon atoms [37], which is correlated to the extent of
nanotube sidewall defects and chemical sidewall function-
alization [38]. The overall appearance of the SWCNTs-O
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Fig. 2. Raman spectra of the GO, rGO, SWCNTs-O and CB.
The GO and rGO curves are superposed and plotted in different
colors (GO = black, rGO = red).
spectrum is compatible with short (1.5 micron average
length) and highly functionalized nanotubes.
The Raman spectrum of CB displays the D and G
bands at 1365 cm¢1 and 1582 cm¢1 respectively. The ID/IG
ratio is 0.97. In a poorly oxidized sample, this is an indica-
tion of the presence of rather ordered graphitic-like
planes [39].
Next, we turned to explore the morphology of SPEs
modified with the carbon nanomaterials via SEM.
In Figure 3 low and high magnification micrographs of
bare SPE, CB-SPE, SWCNTs-O, GO, and rGO SPEs are
compared.
The SEM observation of bare SPE revealed irregularly
shaped micrometric flakes of graphite (Figure 3a and 3b),
randomly orientated within the original ink together with
small particles ascribed to the cross-linking agents. The
SEM micrographs of the SPE modified with CB evi-
denced that the drop coated CB nanoparticles completely
and uniformly cover the working electrode surface, also
originating numerous cauliflower-like agglomerates (Fig-
ure 3c and 3d).
The SPE modified with SWCNTs-O is characterized by
the presence of randomly distributed single short
SWCNTs and, particularly, of sponge-like clusters of
SWCNTs whose formation is strongly promoted by the
very high specific surface area of the nanotubes and their
consequent tendency to agglomerate (Figures 3e–g).
In the case of GO-SPE it is possible to observe a con-
tinuous smooth film on SPE that is so dense and compact
to completely cover the underneath active electrode (Fig-
ure 3h and 3i). This morphology is very different from
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that revealed in the case of reduced GO-SPE which is
constituted by few rGO aggregates (Figure 3j and 3k).
3.2 Electrochemical Characterization
3.2.1 Cyclic Voltammetry Study Using Ferro/ferricyanide
as Probe
For the electrochemical characterization of the nanoengi-
neered electrodes, the cyclic voltammetry method was
used, employing [Fe(CN)6]4¢/3¢ as redox probe. Therefore,
experiments are performed from 0.005 to 1 V/s. The
method of Nicholson [40] is routinely used to deduce the
standard electron transfer rate using the following Equa-
tion 1:
k0 ¼ y ½D0 pv ðnF=RTÞ0:5 ¤ ðDR =DO Þa=2 ð1Þ
where k0 is the standard electron rate constant, y is the
kinetic parameter, Do and DR are the diffusion coeffi-
cients of oxidized and reduced species, v is the scan rate,
F is the Faraday constant, a is the transfer coefficient
(0.5) and n is the number of electrons transferred in the
electrochemical process. y depends on the peak-to-peak
separation (DE). k0, calculated as above, only represents
the apparent rate constant, because no correction related
to the double-layer evolution or iR effects is taken into
account.
The electrochemical response [Fe(CN)6]4¢/3¢ is reported
in Figure 4: the voltammograms acquired at 0.05 V/s are
characterized by a variable estimation of DE. CB-SPE,
SWCNTs-O-SPE, rGO-SPE and GO-SPE, respectively
showed DEs equal to 80, 100, 130 and 120 mV, which is of
a greater significance if compared with the 430 mV ob-
tained using the unmodified SPE.
In a recent work [31], Vicentini et al. evaluated the
electrocatalytic properties of three different carbon black
materials (VXC72R, BP4750, E2000) placed on a glassy
carbon electrode, obtaining DEs (215.3, 237.2 and
452.3 mV) definitely larger than what is obtained by our
CB-SPE using CBN220. DEs obtained for CNTs and gra-
phene modified SPEs are comparable with those reported
in literature [41, 42]. The peak-to-peak separations ob-
tained testing SPEs in presence of this electrochemical
probe, are consistent with k0 reported in Table 1: all the
nanostructured electrodes surface led to a large improve-
ment of the electron transfer for this particular redox
system if compared to bare SPE. CB-SPE is characterized
by a calculated k0 of 4.5 × 10¢3 cm/s, a value similar to the
ones reported in our previous papers [25, 27] and 10–100
fold higher than the ones reported by Mart‡n and Escarpa
[43] in their review on the heterogeneous electron trans-
fer rates for carbon materials based-electrodes.
Furthermore, the voltammetric peak heights (Ip) relat-
ed to each modified-SPE were monitored as a function of
the scan rate and, afterwards, plotted versus the square
root of the scan rate (v0.5). The following slopes (mA/
(mV s¢1)0.5) were obtained: 1.56 (R2 = 0.998), 1.45 (R2 =
0.987), 0.645 (R2 = 0.993), 0.0826 (R2 = 0.997) and 0.388
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Fig. 3. SEM micrographs of bare SPE (a-b), CBNPs-SPE (c-d), SWCNTs-O-SPE (e-g), GO-SPE (h-i) and rGO-SPE (j-k).

(R2 = 0.992), corresponding respectively to, CB-SPE,
SWCNTs-O-SPE, rGO-SPE, GO-SPE and bare SPE.
Each SPE showed a linear response, indicating a diffusion-
al process. Furthermore, associated with Nicholson Equa-
tion 1 is the Randles¢Ševcik Equation 2 [44]
I p ¼ 0:4463 ðF 3 =RTÞ0:5 n1:5 AD0:5 Cv0:5
where Ip is the peak current, F is the Faraday constant, A
is the electrode area, D is diffusion coefficient of the ana-
lyte, v is the scan rate, C is the concentration of the spe-
cies and n is the number of electrons transferred in the
electrochemical process. In the case of semi-infinite diffu-
sion of the probe at the electrode surface, the analysis of
the cyclic voltammetry peaks in terms of the log Ip vs. log
v should consist in a slope equal to 0.5 in order to vali-
ð2Þ date both Equations1 and 2. The slopes of 0.55, 0.52, 0.37,

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Table 1. Electron transfer rate constants, cyclic voltammetric data, amperometric sensitivities and LOD for several compounds at
modified-SPEs.
Bare SPE CB-SPE SWCNTs-O-SPE GO-SPE rGO-SPE
k0 (cm/s) Fe(CN)64¢/3¢ – 0.00452 0.00251 0.00167 0.00247
DE (mV) Fe(CN)64¢/3¢ 430 80 100 130 120
Eox’ (mV) NADH 550 400 440 > 600 530
Sensitivity (mA/M cm2) NADH 4.32 224 281 – 88.5
LOD (mM) NADH 6 1 5 – 5
Eox’ (mV) AA 420 150 90 380 320
Sensitivity (A/M cm2) AA 3.98 278 496 19 203
LOD (mM) AA 7 1 5 3 2
Eox’ (mV) Cysteine 930 580 580 950 700
Sensitivity (A/M cm2) Cysteine 4.82 362 565 10.3 211
LOD (mM) Cysteine 30 1 5 24 6

Fig. 5. Complex plane impedance plots at open circuit potential

Fig. 4. Cyclic voltammograms obtained for 1 mM of ferro/ferri-
cyanide in KCl 0.1 M. All recorded at scan rate of 50 mV/s using
bare SPE (black line) or SPE modified with CB (violet line),
SWCNTs-O (red line), GO (blue line), rGO (green line). Equa-
tions related to the oxidation/reduction reactions of the couple
ferro/ferricyanide are reported in the graph.
using a 5 mM ferro/ferricyanide in 0.1 M KCl for bare SPE
(black line) or SPE modified with CB (violet line), SWCNTs-O
(red line), GO (blue line), rGO (green line). Inset (i) Randles
circuit, (ii) highlight of the complex plane impedance plot at the
high frequency region.

the electrode surface modified with the nanomaterials, by

0.46 and 0.40, related to CB-SPE, SWCNTs-O-SPE,
measuring the value of electron transfer resistance (Rct).
rGO-SPE, GO-SPE and bare SPE, reveal a satisfying cor-
The Rct is estimated according to the diameter of the sem-
relation to the approximated transfer coefficient (0,5);
icircle present at the high frequency region. It represents
thus the redox probe is not trapped within the nanomate-
the difficulty of electron transfer of Fe(CN)64¢/3¢ redox
rial film (no thin-layer effect) and the response follows
probe between the solution and the electrode, thus giving
a semi-infinite diffusion in each analyzed case [45]. The
information on the electrode surface. The electrochemical
values obtained here are in good agreement with the
impedance spectroscopy was performed with the bare
slopes found in other works involving the use of CB,
SPE and with SPE modified with CB, GO, rGO, and
CNTs and graphene as electrode modifiers [31, 45, 46].
SWCNTs-O at open circuit potential (OPC). The fitting
All the reported electrochemical parameters, definitively
of the spectra is done using the equivalent electrical cir-
confirm how these well-defined nano-carbonaceous films
cuit as shown in Figure 5 Inset. It comprises the electro-
possess the required surface structure and electronic
lyte resistance, Re, in series with a parallel combination of
properties to support rapid electron transfer for particular
Rct (interfacial charge transfer resistance), Zw (diffusion
redox systems.
of the analytes in solution and corresponding to Warburg
impedance straight line of the curves) and CPE (Constant
3.2.1. Impedance Spectroscopy Study
Phase Element).
Electrochemical Impedance Spectroscopy provides ex- Figure 5 shows the Nyquist plots for bare SPE, CB-
tremely useful information on the impedance changes of SPE, rGO-SPE, GO-SPE and SWCNTs-O-SPE. The Rct

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Fig. 6. Cyclic voltammograms obtained in phosphate buffer 0.05 M + KCl 0.1 M, pH 7.4 in absence (a) and in presence of NADH (b,
2 mM), ascorbic acid (c, 2 mM) and cysteine (d, 2 mM). All recorded at a scan rate of 50 mV/s using bare SPE (black line) or SPE
modified with CB (violet line), SWCNTs-O (red line), GO (blue line), rGO (green line). Equations related to NADH, ascorbic acid
and cysteine oxidation reactions are reported in the graph, respectively b, c and d.

value for bare SPE is much higher (Rct = 293.9 œ 1.5 kW)
than those for the nanomodified SPE; in the latter case,
the highest Rct is found for GO-SPE (Rct = 68.6 œ 1.2 kW).
The other sensors showed a Rct lower than 100 W, con-
firming the advantage to use the nanomodified SPE.
3.3 Electroanalytical Sensing Towards NADH, Ascorbic
Acid and Cysteine
One of the great advantages to use carbon nanomaterial
to modify electrodes is easily attributable to their wide
potential window and high electrochemically accessible
area: the film physically or chemically deposited onto the
electrode surface, allows the measurement of molecules
with relatively high oxidation potentials. As reported in
Figure 6a, all the modified SPEs have wide operative po-
tential windows when performing a background analysis
in phosphate buffer, ranging approximately from ¢0.3 to
1.0 V (vs. Ag/AgCl), a useful range in voltammetric anal-
ysis. However, it should be noted how SWCNTs-O-SPE,
if compared with other SPEs, is characterized by a larger
capacitive current (Figure 6a) and by the longest time to
reach the stabilization (Figure 7). Here, we studied the
detection of three important electroactive molecules in
clinical and biochemical field: NADH, ascorbic acid and
cysteine.
The electrochemical oxidation of NADH is of great in-
terest because of its requirement in many biosensors
based on dehydrogenase enzymes (about 300 dehydro-
genases are strictly dependent on the coenzyme and its
oxidized form NAD + ). Two critical issues are related to
the high overpotential required to detect target mole-
cules, and to monitor fouling problem.
Here, we studied the oxidation of NADH at the differ-
ent developed SPEs. As illustrated in Figure 6b SPEs
modified with CB and SWCNTs-O show the lowest po-
tentials, respectively 400 and 440 mV. In comparison
rGO-SPE and GO-SPE show peak potentials of at
530 mV and higher than 600 mV, respectively. The over-
potentials achieved here are satisfactory and well-com-
pare with other results found in literature where the same
nanomaterials are used as electrode modifier, highlighting

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Fig. 7. Amperograms obtained in phosphate buffer 0.05 M +
KCl 0.1 M, pH 7.4 to successive additions (25 mM) of NADH (a),
ascorbic acid (b) and cysteine (c) using bare SPE (black line) or
SPE modified with CB (violet line), SWCNTs-O (red line), GO
(blue line), rGO (green line). Applied potential (vs. Ag/AgCl)
was + 600, + 150 and + 400 mV respectively, to detect NADH,
ascorbic acid and cysteine.
the electrocatalytic properties of CB-SPE with respect to
SWCNTs-O, GO, and rGO modified electrodes
[42, 47, 48].
The ascorbic acid plays a very relevant role as antioxi-
dant in the human metabolism and it is important to de-
velop a sensitive platform to detect it. Figure 6c shows as
SWCNTs-O and CB are characterized by the highest cur-
rent peaks related to electrochemical oxidation of ascor-
bic acid at electrode surface and by the lowest potential
peaks, 90 and 150 mV respectively [49–51].
The last electrocatalytic evaluation of modified-SPEs,
using cyclic voltammetry experiments, was performed
with the detection of cysteine, a thiol-containing amino
acid, often involved in enzymatic reactions as a nucleo-
phile. As reported in Figure 6d, CB-SPE and SWCNTs-
O-SPE showed a reduction in the overpotential of cys-
teine oxidation to 580 mV. The oxidation peak potential
of cysteine reached using the bare, GO-SPE and rGO-
SPE, is much higher than that of the CB and SWCNTs-O
SPEs: this implies that the oxidation of cysteine, especial-
ly at SWCNTs-O and CB electrodes, is significantly im-
www.electroanalysis.wiley-vch.de Ó 2015 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
Special Issue ADVANCED MATERIALS
proved. Many systems based on carbon nanomaterials
have been developed to detect these important biomole-
cules and the values shown in this work by CB-SPEs ac-
quire great analytical relevance. CB-SPE shows lower
overpotential than rGO-SPE and GO-SPE. As for
SWCNTs-O-SPE, the analytical performances are similar
to those of CB, also if the comparison is made with works
reported in literature [52, 53], but CB offers a very valua-
ble compromise considering performance and cost.
Based on the above cyclic voltammetric results, the typ-
ical amperometric responses of the different modified
SPEs were examined as a function of different concentra-
tions of NADH, ascorbic acid and cysteine. Ad hoc po-
tentials were applied to achieve the electrooxidation at
the different SPEs: 600, 150 and 400 mV were exploited,
respectively, to detect NADH, ascorbic acid and cysteine.
Figure 7 illustrates the amperograms obtained at the con-
centration range from 0 to 100 mM in phosphate buffer.
As expected, the SWCNTs-O and CB modified-SPEs dis-
play higher current response to the addition of the ana-
lytes compared to the other SPEs.
In Table 1 overpotentials, sensitivities and LOD ob-
tained by using the different carbon nanomaterial as
SPEs modifying tools were reported: although SWCNTs-
O-SPE was capable to yield the best sensitivities, CB-SPE
reached the lowest LOD (calculated as S/N = 3) confirm-
ing its excellent electrocatalytic properties.
4 Conclusions
This work was focused on the comparison among SPEs
modified with CB and with the most used modifiers such
as carbon nanotubes and graphene.
The CB-SPE has shown a better heterogeneous elec-
tron transfer constant in presence of ferricyanide redox
mediator compared to the others modified SPEs. Further-
more, an improvement in terms of reduced overpotential
and/or low background current was found in the case of
ascorbic acid, NADH and cysteine. These behaviors
could be ascribed to the better uniformly CB-coverage of
the working electrode than SWCNTs-O/GO/rGO-SPEs
which displayed cluster structures, as observed from SEM
analysis: a direct consequence is also visible by the calcu-
lated 1 mM LOD in the case of CB-SPE.
Taking into account the CB electrochemical properties,
the cost-effectiveness, the capability to easily obtain
a stable and homogenous dispersion, the CB deserves to
be widely employed in the development of nanomodified
electrochemical sensors. CB dispersion is also character-
ized by a higher stability, compared to other carbona-
ceous dispersions: CB dispersion is stable at least two
weeks while SWCNTs-O and GO dispersions were no
longer homogeneous the day after production (Support-
ing Information, Figure S1). Furthermore, the CB-SPE is
characterized by high storage stability: for at least three
months at room temperature and dry condition, as al-
ready reported [24].
Electroanalysis 2015, 27, 2230 – 2238 2237

Full Paper
Furthermore, the relevance of this study consists on
providing the characterization of different carbonaceous
nanomaterials on SPEs, providing useful tools to develop
sensitive platforms depending on the needs and on the
available resources.
Acknowledgements
F. A. wishes to acknowledge the Minister of Defence,
Aptamer BW project for financial support.
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Received: March 15, 2015
Accepted: April 30, 2015
Published online: July 14, 2015
Electroanalysis 2015, 27, 2230 – 2238 2238