Thin Solid Films 699 (2020) 137875

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Thin Solid Films

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High-performance electrochemical sensor based on molecularly imprinted T

polypyrrole-graphene modified glassy carbon electrode
S.M. Oliveiraa,b, J.M. Luzardoa, L.A. Silvaa, D.C. Aguiara, C.A. Sennaa, R. Verdana, A. Kuznetsova,
⁎
T.L. Vasconcelosa, B.S. Archanjoa, C.A. Achetea, Eliane D'Eliab, J.R. Araujoa,
a
Materials Metrology Division, National Institute of Metrology, Quality and Technology (Inmetro), Av. Nossa Senhora das Graças, 50, CEP 25250-020, Duque de Caxias,
Brazil
b
Instituto de Química, Universidade Federal do Rio de Janeiro, Av. Athos da Silveira Ramos, 149, Cidade Universitária, CEP 21044-020, Rio de Janeiro, Brazil

A R T I C LE I N FO A B S T R A C T

Keywords: Molecularly imprinted polymers are pushing the limits of sensing capabilities in electrochemical sensors. As a
Graphene result, these sensors are driving the improvement of chemical species detection with applications in pharma-
Molecularly imprinted polymer ceutical, chemical and food industries as well as showing potential for bioanalytes sensing for applications in
Polypyrrole medical diagnostics. In particular, molecularly imprinted polymer-graphene sensors have great potential to
Ascorbic acid
increase the selectivity, sensitivity and detectability of electrochemical sensors for organic molecules determi-
Electrochemical sensor
nation, especially in the presence of interferers. In this work, we tested three different coatings on glassy carbon
electrode for the purpose of ascorbic acid quantification: graphene, molecular imprinted polypyrrole-graphene
and non-imprinted polypyrrole-graphene. Among the prepared electrodes, non-imprinted and molecularly im-
printed polypyrrole on graphene, exhibited the highest sensitivity to ascorbic acid (6.14 and 5.87 μA/mmol L−1,
respectively), although their detection limit (0.56 and 0.10 mmol L−1, respectively) was poore than the un-
coated graphene one (0.02 mmol L−1). Besides that, graphene modified glassy carbon electrode showed better
response for repetitive uses. However, in the case of a unique measurement to detect the presence of ascorbic
acid in a solution containing interferers, as uric acid and dopamine, molecular imprinted polypyrrole-graphene
electrode exhibits better response and selectivity in addition to, an electrocatalytic effect.

1. Introduction separation of the individual contribution to the overall electrochemical

Ascorbic acid (AA, vitamin C) is a vitamin easily found in vege-
tables, legumes and fruits. As a powerful antioxidant, ascorbic acid is
widely used in the food industry to preserve flavor and the natural color
of many foods, processed fruits and vegetables and dairy products
[1,2]. Recent studies have shown the importance of AA in the protec-
tion against chronic diseases caused by oxidative stress [3]. Hence, it is
important to develop simple, rapid, selective and sensitive analytical
methods for the detection of AA. Over the past years, great progress has
been made in the development of electrochemical AA sensors [4-11],
however many analytical challenges remained to overcome in order to
make these sensors cost-competitive for applications in pharmacolo-
gical analysis as routine clynical diagnoses. One of such challenges is
the elimination of the interference of dopamine (DA) and uric acid
(UA), normally present in biological fluids, for the electrochemical
determination of AA. In the case of these three analytes, the respective
oxidation peak potentials are close to each other making difficult the
signal. Besides, the oxidation products of all components are adsorbed
on the electrode surface compromising its subsequent utilization.
Therefore, the development of nanomaterials to improve electrode
sensitivity and, principally, selectivity for the determination of AA, in
the presence of interfering compounds, has been a subject of number of
studies in the last years [5-8,12-15].
Electrochemical sensors, based on graphene sheets, possess the
improved sensitivity and detectability for the probe of organic mole-
cules and biological bodies due to special graphene properties, as high
electrical conductivity, mechanical resistance and high surface area
[16]. The selectivity of these sensors can be further improved by ap-
plying molecular imprinting technology. MIPs are synthetic polymers
that exhibit high molecular recognition properties for a template mo-
lecule by acting similarly to enzyme-substrate or antigen-antibody
systems in the molecular recognition process of a substance chosen as a
template [12,17-20]. Polypyrrole (PPy) is an instrinsically conducting
polymer suitable for molecular imprinting due to the combination of

⁎
Corresponding author.
E-mail address: jraraujo@inmetro.gov.br (J.R. Araujo).

https://doi.org/10.1016/j.tsf.2020.137875
Received 30 May 2019; Received in revised form 12 February 2020; Accepted 19 February 2020
Available online 21 February 2020
0040-6090/ © 2020 Elsevier B.V. All rights reserved.
S.M. Oliveira, et al.
electrochemical redox activity and good electrical conductivity (2 to
100 S cm−1) [21], that provide a good recognition capacity towards
different target molecules [22].
There is a growing number of reports in the literature on graphene
usage as an electrode modifier in polypyrrole-molecular imprinting
polymers (PPy-MIP), to detect different organic molecules [13-
15,23,24]. Chein et al. [7] prepared a graphene-based hybrid film
(obtained by rGO electroplating) and polypyrrole (superoxidized and
electropolymerized) on a glassy carbon electrode using cyclic voltam-
metry aiming dopamine detection. The modified electrode exhibited
excellent catalytic activity for DA and UA but repelled the ascorbate
anion under physiological pH conditions. The response to dopamine
was linear in the concentration range from 2.0 μM to 160 μM, the de-
tection limit (LOD) was 0.5 μmol L−1, even in the presence of ascorbic
acid and uric acid.
The examples of similar studies include: graphene and PPy-MIP
modified glassy carbon electrode for determination of trimethoprim
(LOD = 1.3 × 10−7 mol L−1) [24], MIP with incorporated graphene
oxide for determination of quercetin (LOD = 4.8 × 10−8 mol L−1)
[23], PPy-MIP synthesized on gold nanoparticles and graphene mod-
ified glassy carbon electrode for estradiol detection
(LOD = 1.0 × 10−9 mol L−1) [14], glassy carbon electrode modified
with graphene, gold nanoparticles and molecularly imprinted over-
oxidized PPy for electrochemical determination of dopamine
(LOD = 1.0 × 10−7 mol L−1) [13], graphene modified carbon paste
electrode for ascorbic acid determination (LOD = 7.0 × 10−8 mol L−1)
[15], to mention a few.
In this work, we develop a sensor for AA detection: a glassy carbon
electrode (GCE), simultaneously modified by electrochemically ex-
foliated grafene, named as EG, and molecularly imprinted PPy to create
a highly selective and sensitive electrode for AA detection in the pre-
sence of interferers as DA and UA.
2. Material and methods
2.1. Reagents and apparatus
The reagents used in this work were of analytical grade and all
solutions were prepared using water purified by a Milli-Q Millipore
system (resistivity ≥18 MΩ cm2). Ammonium sulfate (≥99.0%), n,n’-
dimethylformamide (99%), L-ascorbic acid (>99.5%), lithium per-
chlorate (> 99.99%) and pyrrole (98%) were obtained from Sigma-
Aldrich (Rio de Janeiro, Brazil). Graphite sheet of 0.25 mm thick was
obtained from Alfa Aesar (Tewksbury, United States).
All electrochemical measurements were performed using an Autolab
potentiostat, model PGSTAT 204 (Metrohm Autolab B.V., Utrecht, The
Netherlands) and NOVA 1.11 software for data acquisition. The
working electrode was a graphene modified glassy carbon (diameter of
3 mm), a platinum wire as the counter electrode and Ag/AgCl (sat. KCl)
electrode as the reference electrode.
2.2. Preparation of EG
In view of the challenges regarding large-scale production of large
area defect-free graphene sheets, Parvez et al. [25] has been reported a
graphene synthesis method based on the electrochemical exfoliation of
graphite using a platinum wire as the counter electrode and a graphite
foil as the working electrode. A solution of (NH4)2SO4 0.1 mol L−1 was
used as a carrier electrolyte. The potential of 10 V, applied between the
graphite and platinum electrodes, during approximately 30 min, re-
sulted in complete exfoliation of graphite electrode in electrolyte.
Afterwards, EG sheets were separated by filtration of electrolyte and
dispersed in dimethylformamide (DMF) for 15 min. Four microliters of
EG dispersion (10 mg mL−1) were dripped onto the surface of the glassy
carbon electrode and then, the graphene-modified electrode was dried
in an oven at 50 °C temperature.
2
Thin Solid Films 699 (2020) 137875
2.3. EG characterization
X-ray photoelectron spectroscopy (XPS) analyses of EG were per-
formed in an ultra-high vacuum chamber (pressure: 10−7 Pa) (Scienta
Omicron GmbH, Taunusstein, Germany) using an Al (Kα = 1486.7 eV)
X-ray source operating at the anode voltage of 15 kV and filament
current of 20 mA. Survey spectra were obtained with analyzer pass
energy of 160 eV and step of 1 eV. The high-resolution spectra (C 1s)
were obtained with 20 eV analyzer pass energy.
Raman spectroscopy analyses of EG were carried out on a Witec
Alpha 300 spectrometer (Ulm, Germany), with a 532 nm laser line in a
backscattering configuration using a microscope with a 100 × objec-
tive. The laser power was kept under 0.5 mW to avoid local heat and
damages to samples. All spectra were acquired using 10 s integration
time and an average of 10 accumulations. The final spectrum was ob-
tained by averaging three measurements taken at random points to
evaluate the graphene homogeneity.
X-ray diffraction (XRD) analyses of EG were performed on a D8-
Focus (Bruker, Massachusetts, United States) diffractometer using Ni-
filtered Cu-Kα radiation (λ = 1.5406 Å) with the step of 0.02° and a 2θ
scanning range from 5° to 40°. A thin film sample was prepared by drop
drying the aqueous suspension of graphene onto Si wafers.
2.4. Fabrication of GCE-EG-MIP sensor
MIP for AA determination was prepared on the EG covered glassy
carbon electrode following the procedure of Özcan et al. [9] using
0.1 mol L−1 LiClO4, 25 mmol L−1 pyrrole and 10 mmol L−1 AA solu-
tion. The EG modified electrode was immersed in this solution where
the electropolymerization was carried out by cyclic voltammetry in the
potential range between −0.60 and +0.80 V, during seven cycles, with
the scan rate of 100 mV s−1. After the completion of electro-
polymerization, AA was extracted in a phosphate-buffered saline (PBS)
solution 0.05 mol L−1, with gentle agitation for 20 min. The molecule
rebinding was performed in a 0.5 mmol L−1 AA solution for 20 min.
Different concentrations of ascorbic acid were measured by differ-
ential pulse voltammetry (DPV) using a three electrode cell, in
0.1 mol L−1 KCl + 0.05 mol L−1 PBS at pH 7.2, in the potential range
between −0.40 and 0.60 V, step potential of 10 mV, modulation am-
plitude of 25 mV and scan rate of 20 mV s−1. All electroanalytical
measurements were carried out at room temperature and in triplicate in
order to plot an analytical curve for each fabricated electrode.
2.5. Sensor characterization
Scanning Electron Microscopy (SEM) of the fabricated electrodes
was performed using a Field Emission Scanning Electron Microscope
(FEG-SEM-FIB), model Helios NanoLab 650 (FEI-Thermo Fisher
Scientific, Hillsboro, United States), operated at 1 kV, 25 pA. The
transmission electron microscopy (TEM), and high-resolution TEM
(HRTEM) analysis were carried out on a Titan 80–300 equipped with
EDS-EDAX Si(Li) detector (FEI-Thermo Fisher Scientific, Hillsboro,
United States) using an acceleration voltage of 80 kV to prevent the
sample from electron irradiation damaging effects.
2.6. Preparation of sample for TEM analysis
The sample preparation procedure for TEM analysis was performed
covering the gold TEM grid with the EG dispersed in DMF solution. Four
microliters of EG dispersion (10 mg mL−1) were dripped onto the
surface of a gold grid and after, it was attached to the glassy carbon
electrode with graphite glue and was dried in an oven at 50 °C for
10 min. The MIP for AA determination was prepared on the gold TEM
grid following the same procedure employed for the fabrication of GCE-
EG-MIP sensor. For AA extraction, the grid was immersed in PBS so-
lution with gentle agitation for 20 min.
S.M. Oliveira, et al.
3. Results and discussion
3.1. Sensor fabrication by MIP electropolymerization
In this work, we studied four different sensors, a glassy carbon
commercial electrode (named as GCE) and three graphene-modified
GCE: EG modified GCE (named as GCE-EG), non-imprinted polypyrrole
on EG modified GCE (named as GCE-EG-NIP) and AA molecularly im-
printed polypyrrole on EG modified GCE (named as GCE-EG-MIP).
The polymerization of pyrrole monomers was achieved through
electrochemical oxidation process at the GCE surface coated by EG.
Fig. 1 shows the cyclic voltammograms recorded during the electro-
polymerization of pyrrole (25 mmol L−1). Fig. 1a shows GCE-EG-NIP
and Fig. 1b shows GCE-EG-MIP. The formation and growth of the
polymer film can be seen in both electrodes, GCE-EG-NIP and GCE-EG-
MIP, by the increase in the intensity of the oxidation and reduction
peak. During the NIP growth (see inset in Fig. 1a), a broad oxidation
peak was observed at about −0.1 V and reverse cathodic peak was seen
at approximately 0.15 V. Besides the pyrrole oxidation peak, the ap-
pearance of an oxidation peak in Fig. 1b at about 0.4 V indicates the
incorporation of AA template molecules in the polymeric chain formed
on GCE-EG. AA molecules that diffuse towards the surface of the GCE-
EG electrode during the electropolymerization process are trapped in-
side the polymer matrix due to the non-covalent interactions between
template molecules and the pyrrole units [20]. This process is able to
create a higher number of recognition sites than a non-electroactive
template [9].
3.2. Characterization of graphene and MIP-graphene sensor
Structural, chemical and morphological characterizations of EG
were performed to elucidate its structure, the type of the electron hy-
bridization of carbon bonds, purity and functionalization groups using
Raman, XPS and XRD techniques. In the case of EG-MIP electrode, its
chemical and phase compositions were elucidated using Energy
Dispersive Spectroscopy (EDS) and HRTEM.
XRD patterns of the EG sample show the broad peak with the pro-
nounced maximum at 26.5° 2θ, characteristic of (002) Bragg reflection
of graphite (see Fig. 2a) [26]. The strong broadening of this peak may
reflect the presence of few layered coherently scattering graphene do-
mains randomly oriented and having a dispersed interplanar spacing.
The negative correlation between the average lateral size of the initial
graphene sheets and the broadening of the XRD peak has been estab-
lished by Castro et al. [26]. However, this effect, most probably, is not a
dominant contribution to the XRD peak broadening. The crumpled and
wrinkled character of graphene sheets observed in microscopy studies
(see Fig. 2d to –g) suggests the microstrains along the stacking direction
and stacking faults to be the main source of this effect.
XPS analysis provides valuable information about the oxidation
degree of EG. Fig. 2b shows the XPS spectra of the samples in the C1s
core level binding energy region. C1s spectrum was decomposed in six
peak components centered at the following energies: 284.6 eV (sp2
C]C graphite carbon skeleton, 53% of total peak area), 285.5 eV (sp3
3
Thin Solid Films 699 (2020) 137875
Fig. 1. Sensor fabrication by cyclic voltammetry recorded during the
electropolymerization of pyrrole (25 mmol L−1). Current vs. potential
curves were measurements during: (a) non-imprinted PPy film de-
position and (b) molecularly imprinted PPy film deposition, to gen-
erate GCE-EG-NIP and GCE-EG-MIP, respectively. The data of cyclic
voltammetry were collected with 100 mV s−1 scan rate, 0.1 mol L−1
LiClO4 supporting electrolyte for seven cycles of forward and reverse
potential scans.
CeC graphite carbon skeleton, 21% of total peak area), 286.7 eV
(CeOH and CeOeC, hydroxyl and epoxy groups, 8% of total peak
area), 287.4 eV (C]O, carbonyl, 8% of total peak area), 288.8 eV
(CeOOH, carboxyl, 8% of total peak area) and 291.4 eV (π-π* shake-up
satellite peak, 2% of total peak area). XPS data show that EG has lower
oxidation degree than a typical graphene oxide material. EG shows sp2/
sp3 ratio of ~1.5 while the reported GO sp2/sp3 ratio is ~0.2 [26]. The
shake-up satellite peak component, typical of the delocalized electrons
in π aromatic systems [27], is also an indicative that the sp2 hexagonal
structure is preserved, in part, in EG structure. Fig. 2c shows the Raman
spectrum of EG. The band at 1580 cm−1, known as the G-band, re-
presents the in-plane stretching of carbon atoms in graphitic structures
[28-30]. The presence of disorder or defects in the lattice, such as va-
cancies, oxygen containing functional groups and the adsorption of
molecules on the graphitic structure surface, are revealed by the pre-
sence of the D-band, which is positioned near 1350 cm−1 [30,31]. The
bands positioned near 2700 cm−1 and 2900 cm−1 are known as a
contribution of second order of D band (named as 2D) and a combi-
nation of G and D bands (named as D+G), respectively [32]. The D and
G bands parameters of the EG Raman spectra were fitted using Lor-
entzian functions. Table 1 presents the mean values and uncertainties of
the fitting parameters: frequency (F), FWHM (W) of both D and G
bands, and relative intensity I(D)/I(G). The width of a Raman band can
be directly correlated with the degree of disorder in a material [33].
Although GO presents broader D and G bands than EG, with point de-
fects caused by chemical oxidation and ultrasound exfoliation pro-
cesses, it has a lower ID/IG ratio than the EG sample because the
electrochemical process causes further edges defects in the graphene
sheets contributing to the increase of the total defect density [34].
The surface morphologies of the electrodes in its fabrication steps
were examined using SEM and TEM. SEM image of GCE-EG electrode,
Fig. 2d, shows the typical crumpled and wrinkled graphene sheets
structure before PPy-MIP deposition. In Fig. 2e, less pronounced step
edges of graphene and uneven imprinted PPy-MIP layers were clearly
observed on the GCE-EG-MIP electrode surface, confirming the pre-
sence of PPy-MIP coating on graphene layer. Both electrodes, GCE-EG
and GCE-EG-MIP, have rough surface that results in a large surface
area, which, in turn, favors the higher number of active sites for the
recognition of the target analyte. Fig. 2f shows a graphene sheet with
ultrafine morphology, with folds at their edges. The insets in Fig. 2f
shows a HRTEM and fast Fourier transformed, filtered and inverted
images of the selected regions confirming the typical “honeycomb”
atomic structure of graphene. Fig. 2g shows EG-MIP micrograph with a
seemingly thicker sheets and rougher sheets edges as compared to the
graphene flakes (Fig. 2f). Remarkably, graphene sheets in all obtained
micrographs were fully covered by the MIP confirming the efficacy of
the employed method of polymer synthesis by molecular imprinting on
EG. The collected EDS spectrum (see inset in Fig. 2g) acquired at the
dashed box marked region in Fig. 2g shows the presence of N atoms,
which is the signature of MIP. In addition, the electron diffraction
pattern of the same region, in Fig. 2g, indicates the preservation of
hexagonal graphene structure after the MIP deposition.
Fig. 3 shows the EG-MIP flake before (Fig. 3a) and after (Fig. 3b) the
S.M. Oliveira, et al. Thin Solid Films 699 (2020) 137875

Fig. 2. Chemical and structural characterization of EG, where: (a) X-ray diffraction pattern, (b) C1s high-resolution XPS spectrum, and (c) Raman spectrum. SEM
images of electrodes: (d) GCE-EG and (e) GCE-EG-MIP surfaces, (f) TEM of the EG flake. The insets show the HRTEM image of the EG flake and the inverted FFT image
of the selected region; (g) TEM micrograph of the EG-MIP flake. The EDS spectrum in the top inset was acquired at the dashed box area and indicates the elemental
polymer composition of the observed MIP layer. The bottom inset is an electron diffraction (SAED) of this selected-area, which is addressed as a graphene layer
embedded with MIP.

AA extraction in a PBS solution. These micrographs provide evidences the next section, provided the direct confirmation of the improvement
of a heterogeneous morphology of EG-MIP flake after the AA extraction in the overall performance of EG-MIP modified electrode.
as compared to the one before the AA extraction. The differences in the
electrochemical performance between studied electrodes, presented in

Table 1
Mean values and expanded uncertainties of fitting parameters of the D and G bands for EG sample and comparison with graphene oxide (GO) parameters [35].
Sample F (D) cm−1 W (D) cm−1 F (G) cm−1 W (G) cm−1 I(D)/I(G)

EG 1346.4 ± 1.6 103.4 ± 5.1 1591.0 ± 1.3 74.8 ± 3.5 1.1 ± 1.1
GO 1362.8 ± 9 110 ± 3 1584.8 ± 9 79.5 ± 1 0.86 ± 1

4
S.M. Oliveira, et al. Thin Solid Films 699 (2020) 137875

Fig. 3. Micrographs of TEM samples of EG-MIP (a) before and (b) after the AA extraction (see the details for the samples preparation in the Section 2.6).

Fig. 4. DPV responses of bare GCE and GCE modified elec-

trodes in different AA solutions (concentrations ranging be-
tween 0.5 and 8 mmol L−1), measured in
0.1 mol L−1KCl + 0.05 mol L−1 PBS at pH 7.2 in the po-
tential range between −0.40 and 0.60 V, where: (a) GCE, (b)
GCE-EG, (c) GCE-EG-NIP and (d) GCE-EG-MIP. The analytical
curve of DPV response vs. AA concentration and the re-
spective R2 coeficients are shown in the inset of each plot.

. . . . . . . .

. . . . . . . .

3.3. Evaluation of sensors response studied electrodes (GCE, GCE-EG, GCE-EG-NIP and GCE-EG-MIP) in
order to compare sensitivity and selectivity of the prepared electrodes.
Different concentrations of ascorbic acid (from 0.1 to 8 mmol L−1) The comparison of the modified electrodes shows that the GCE-EG-NIP
were analyzed by differential pulse voltammetry at the surface of GCE and GCE-EG-MIP exhibited the highest sensitivity to AA (6.14 and
(Fig. 4a), GCE-EG (Fig. 4b), GCE-EG-NIP (Fig. 4c) and GCE-EG-MIP 5.87 μA/mmol L−1, respectively), although their LOD (0.56 and
(Fig. 4d) electrodes. The response to AA was almost linear in the whole 0.10 mmol L−1, respectively) was poore than the one of GCE-EG
studied concentration range for GCE and EG electrodes (from (0.02 mmol L−1). Furthermore, the graphene seems to promote an
0.1 mmol L−1 to 8 mmol L−1), while the linearity of the GCE-EG-NIP electrocatalytic effect, shifting the AA oxidation peak to lower poten-
and GCE-EG-MIP was observed from 2.0 mmol L−1 to 8 mmol L−1. tials when compared to the unmodified GCE, even after NIP and MIP
Table 2 shows the electrochemical parameters evaluated from the coatings. The improved sensitivity of GCE-EG-NIP and GCE-EG-MIP as
regression parameters of the analytical curves (Fig. 4) for the four compared to GCE-EG (Fig. 2d) may be attributed to the increase in
electrode roughness, as observed by the SEM image (Fig. 2e), which
Table 2 promotes a significant increase of its surface area [20]. The improved
Average electrochemical parameters of GCE, GCE-EG, GCE-EG-NIP and GCE- response of GCE-EG-NIP than GCE-EG-MIP is due to the higher rugosity
EG-MIP electrodes evaluated from DPV for 5 mmol L−1 ascorbic acid con- of GCE-EG-NIP electrode, since that non-imprinted polypyrrole cover
centrations. higher area on electrode surface than molecularly imprinted one. The
Electrode Sensitivity μA/mmol L−1 LOD (mmol L−1)* R2 increase in the effective surface area and in the number of active sites,
which leads to enhanced electrochemical activity, was already reported
GCE 4.20 0.03 0.995 in earlier studies that used graphene as coating for glassy carbon
GCE-EG 5.11 0.02 0.995
electrodes [36,37].
GCE-EG-NIP 6.14 0.56 0.982
GCE-EG-MIP 5.87 0.10 0.996 The electrooxidation of AA, in the presence of the possible inter-
fering substances, like uric acid and dopamine, under similar con-
⁎
LOD = 3.3(Sy/S), where Sy is the standard deviation of the response centrations, at pH 7.2, was also studied in order to compare the per-
(which is based on the standard deviation of y-intercepts of regression lines) formance of the GCE-EG and GCE-EG-MIP. These substances are
and S is the slope of the analytical curve.

5
S.M. Oliveira, et al.
Fig. 5. Normalized peak current versus applied potential for AA, UA and DA (2 mmol L−1) as well as the AA response in the presence of both interferes, UA and DA for
(a) GCE-EG and (b) GCE-EG-MIP electrodes.
Table 3
Effect of UA and DA as interferers in AA DPV response (2 mmol L−1 concentration) using GCE-EG and GCE-EG-MIP electrode.
GCE-EG
Analyte Position (V) Peak height (μA)
AA 0.18 10.29
UA 0.26 2.38
DA 0.25 12.21
AA + UA + DA 0.26 22.04
normally present in biological fluids and may interfere in the AA de-
termination using conventional methods of analysis.
The dependence of the oxidation current of each interferer, at the
fixed AA concentration of 2 mmol L−1, was investigated using DPV.
Fig. 5 shows the normalized plot of current vs. potential for AA, UA and
DA measured individually with GCE-EG (Fig. 5a) and GCE-EG-MIP
electrode (Fig. 5b), as well as the AA response in the presence of both
interferers, UA and DA, in the solution.
The potentials of AA oxidation towards DA, UA and AA are rela-
tively close to each other (see Table 3), which makes it difficult to
detect AA in the presence of these interferers. As expected, GCE-EG-MIP
showed higher response toward AA (16.86 μA peak current) than GCE-
EG electrode (10.29 μA peak current) due to its higher selectivity for
this molecule. Besides that, AA oxidation peak potential was shifted
from 0.18 eV to 0.04 eV when GCE-EG-MIP electrode was the probe due
to a strong electrocatalytic effect promoted by the interaction between
AA molecules and MIP active sites. This result corroborates with the
fact that GCE-EG-MIP can recognize AA molecules better than GCE-EG
via preferential selection of AA functional groups by size, shape and
charge distribution.
It is worthwhile to discuss in conclusion the repeatability char-
acteristic of the studied sensors. The determination of the analytical
curves of current peak height vs. AA concentration (see Fig. 4) involved
a reuse of each electrode under repeated, matching conditions. All
sensors exhibited an impaired response in subsequent measurements
reflected in more and more reduced peak currents. This phenomenon
can be attributed to the adsorption of reaction products on the electrode
surface with the consequent decrease in the number of the active sites
on the electrodes’ surfaces. In the case of MIP-containing electrode, this
effect appeared to be more pronounces as compared to other electrodes
due to a much stronger chemical interaction between the AA oxidation
product and the electrode's active sites.
4. Conclusion
Molecularly imprinted polypyrrole is a powerfull tool to improve
sensitivity and selectivity of glassy carbon electrodes. MIP template has
demonstrated high sensitivity and selectivity for AA detection even in
the presence of uric acid and dopamine as interferers. Graphene by it-
self, has the ability to improve the sensitivity of glassy carbon elec-
trodes due to its superior electrical conductivity and high surface area.
Thin Solid Films 699 (2020) 137875
GCE-EG-MIP
FWHM (eV) Position (V) Peak height (μA) FWHM (eV)
0.24 0.04 16.86 0.16
0.09 0.42 3.31 0.20
0.26 0.32 6.16 0.23
0.28 0.30 16.50 0.25
In the case of routine analysis, requiring several evaluations, graphene-
NIP is more appropriate than graphene-MIP coated electrodes because
MIP strongly adsorbs oxidation products on its surface. However, in the
case of the need of an analytic determination, in the presence of in-
terferes, MIP has better performance than graphene and graphene-NIP
electrodes due to its specific recognition capacity. MIP acts as a syn-
thetic receptor capable of binding to the AA molecule, which fits into
the binding site with high affinity and specificity due to electrostatic
interactions, hydrogen bonds and Van der Waals forces.
CRediT authorship contribution statement
S.M. Oliveira: Data curation, Writing - review & editing. J.M.
Luzardo: Data curation, Formal analysis. L.A. Silva: Data curation,
Formal analysis. D.C. Aguiar: Data curation, Formal analysis. C.A.
Senna: Data curation, Formal analysis. R. Verdan: Data curation,
Formal analysis. A. Kuznetsov: Data curation, Formal analysis. T.L.
Vasconcelos: Data curation, Formal analysis. B.S. Archanjo: Data
curation, Writing - review & editing. C.A. Achete: Data curation,
Writing - review & editing. Eliane D'Elia: Data curation, Writing - re-
view & editing. J.R. Araujo: Data curation, Writing - review & editing.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influ-
ence the work reported in this paper.
Acknowledgments
S. M. de Oliveira, L.A. da Silva, J.R. Araujo, E. D'Elia, B.S. Archanjo
and R.C. Verdan would like to thank FAPERJ: E-26/202.746/2018/
239332; E-26/202.629/2017; E-26/200.319/2017; (Foundation for
Research Support of the State of Rio de Janeiro) and J.R. Araujo, B.S.
Archanjo, C.A. Achete and L. Andrade would like to thank the CNPq:
311900/2017-8 (Conselho Nacional de Desenvolvimento Científico e
Tecnológico) for the fellowship support.
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