Brazilian Journal of Analytical Chemistry

2021, Volume 8, Issue 32, pp178–197

doi: 10.30744/brjac.2179-3425.AR-31-2021

ARTI CL E

Experimental Design to Enhance Dopamine

Electrochemical Detection Using Carbon Paste
Electrodes
Soraya Adriane Blum1 , Felipe Zahrebelnei1 , Noemi Nagata2 , Valtencir Zucolotto3 ,
Luiz Henrique Capparelli Mattoso4 , Christiana Andrade Pessoa1 , Jarem Raul Garcia1 ,
Karen Wohnrath1*
1
Departamento de Química, Universidade Estadual de Ponta Grossa, Av. Carlos Cavalcanti, 4748,
ZC 84030-900, Ponta Grossa, PR, Brazil
2
Departamento de Química, Universidade Federal do Paraná, Centro Politécnico, ZC 81531-980,
Curitiba, PR, Brazil
3
Instituto de Física de São Carlos, Universidade de São Paulo, Av. Trabalhador São-carlense, 400,
Parque Arnold Schimidt, ZC 13566-590, São Carlos, SP, Brazil
4
Laboratório Nacional de Nanotecnologia Aplicada ao Agronegócio (LNNA), Embrapa Instrumentação,
ZC 13560-970, São Carlos, SP, Brazil

electrodes to be used in the fast determination of

neurotransmitters, either in commercial drugs or in biological
samples. Determination of dopamine (DA), for example, is
of great importance since the lack of this neurotransmitter
is related to many neurological disorders, including

as DA detection studies of paste electrodes incorporating

(Eeonomers®), pristine carbon black, graphite, and carbon

nanotubes. Emphasis is put on the smaller particle size and
®
materials,

Eeonomers®

ability of distinguishing DA from its natural interferent,

electrode composition (proportion of Eeonomers®) on the electrochemical sensing of DA. A greater

Cite: Blum, S. A.; Zahrebelnei, F.; Nagata, N.; Zucolotto, V.; Mattoso, L. H. C.; Pessoa, C. A.; Garcia, J. R.; Wohnrath, K.
Experimental Design to Enhance Dopamine Electrochemical Detection Using Carbon Paste Electrodes. Braz. J. Anal. Chem.,
2021, 8 (32), pp 178–197. doi: http://dx.doi.org/10.30744/brjac.2179-3425.AR-31-2021

Submitted 01 March 2021, Resubmitted 08 June 2021, Accepted 10 June 2021, Available online August 2021.

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Braz. J. Anal. Chem., 2021, 8 (32), pp 178–197.

to be promising tools to perform quick, cheap and sensitive detection of this neurotransmitter in bioanalytical
systems.

Keywords: carbon paste electrode, carbon black, intrinsically conducting polymers, electroanalysis,
electrocatalysis, response surface, factorial design.

INTRODUCTION

The use of carbonaceous materials for constructing CPEs has been enabling the production of excellent

comparison to graphite-based CPEs.

for applications as electroconductive additive in hydrogen storage devices, and as cathode materials in

investigated due to their excellent electrical conductivity and presence of several defect sites. Those
properties contribute to faster electron transfer kinetics on the electrodic material, providing interesting
abilities

and CB nanomaterial.
DA is a biological amine that has excitatory and inhibitory activity in postsynaptic membranes, performing

samples, DA concentration ranges from 10 to 10-9 mol L-1

to several neurological disorders, such as Parkinson’s disease, Alzheimer’s disease and schizophrenia,
among others. This neurotransmitter is also used as an intravenous resuscitation medication in hospitals,

The method commonly used for DA analysis is high-performance liquid chromatography (HPLC).

such as the need for expensive equipment, demand for trained technicians, complex sample preparation

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Blum, S. A.; Zahrebelnei, F.; Nagata, N.; Zucolotto, V.; Mattoso, L. H. C.; Pessoa, C. A.; Garcia, J. R.; Wohnrath, K.

procedures and the impossibility of in situ analysis. In contrast, electrochemical methods can overcome

cost. Furthermore, the instrumentation required for electroanalytical determinations is simple and can be
in situ

stand out due to their oxidation potential being close to the DA one and to the fact that their concentrations
being hundreds of thousands of times higher than the concentration of DA in physiological environment.
Additionally, the oxidation products of these compounds can be potentially absorbed or electropolymerized
on the surface of the electrodes, reducing sensors reuse and reproducibility. Therefore, for a DA sensor to

The use of multivariate optimization tools, such as factorial design and response surface methodology

of such approaches has been leading to enhanced detection performances for several sensing devices.

made via in situ deposition of intrinsically conducting polymers. These materials have found applications

conductivity of the material and in the number of surface active sites due to the presence of alternating

sensitive, selective and feasible electrochemical sensors to perform DA determination in real samples.

MATERIALS AND METHODS

Chemicals

ca. 1400 m2
ca. 570 m2 ) and CB 3

ca. 390 m2/g

3
). Details on synthesis, characterization and applications of Eeonomers® can

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Braz. J. Anal. Chem., 2021, 8 (32), pp 178–197.

Preparation of the carbon paste electrodes

Electrochemical measurements

by using cyclic voltammetry, since this technique is the most suitable for a profound characterization in
relation to the kinetic aspects of electrochemical reactions occurring in the electrode/solution interface.
vs Ag/AgCl, at scan rates of
10 mV s and 90 mV s . NaCl 0.5 mol L
-1 -1 -1 -1

µmol L-1. All

Experimental Design and Response Surface Methodology

and pH of the
®

supporting electrolyte. Using these parameters, optimization based on a 3 factorial design and response
2

Eeonomers®
Eeonomers®

®
(i)

the conductivity of the system by polymer doping; (ii)

Subsequently, randomized electrochemical analyses in presence of 50 µmol L-1

potential range from 0 to 800 mV vs Ag/AgCl, at 30 mV s-1

DA Determination in Commercial Drugs

mg of Dopamine Hydrochloride. Aliquots of the commercial sample (20 µ

of DA standard stock solution (1 x 10-2 mol L-1
using a previously prepared electrode containing the Eeonomer® KP20 (KPPE) and NaCl as supporting
electrolyte.

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Experimental Design to Enhance Dopamine Electrochemical Detection Using Carbon Paste Electrodes

RESULTS AND DISCUSSION

composes all electrodes. The amount of binder depends on the granulation of the carbon-based material
and this proportion can cause increment of background current. According to Malha et al., 2013, as CB

As Eeonomers®

®
due to

®
related to the binder. Trying to avoid the use of higher amounts of mineral oil, KP20

employed as optimal electrode compositions.

solution, NaCl 0.5 mol L-1 (pH = 4.0) at 30 mV s-1

Figure 1. Cyclic voltammograms for the KPPE, KPyPE,

KP0PE, GRPE and CNTPE, at = 30 mV s-1. Supporting
electrolyte: NaCl 0.5 mol L-1 at pH 4.0. For better visualization,
the voltammograms for the KPyPE, KP0PE, GRPE and CNTPE
electrodes are highlighted in an inset in the Figure.

the material employed for electrode fabrication. It is interesting to observe that neither the KPPE nor the

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It is interesting to notice that the capacitive current for the KPPE is 23 times higher than for its counterpart

to the former material. This can be understood considering that under acid doping PANI conductivity is

is 2.5 higher than the area for the KP20, fact that corroborate the occurrence of pseudocapacitive process
®
.
Eeonomers ®

electrochemical behavior aiming the construction of electrochemical sensors (S2).

®
KP0PE, KPPE and KPyPE in the presence of
®
is related to

Eeonomers®

separation ( Ep) ca. 83 mV and an anodic and cathodic peak current ratio for the process (Ipa/Ipc) of ca.

®
(Figure 2.a). The parameters obtained from the measured cyclic voltammetry for the

a b
Figure 2. a) Cyclic voltammograms for the electrodes KP0PE, KPPE and KPyPE, and b) Cyclic
voltammograms for the electrodes GRPE, KP0PE e CNTPE, in 50 µmol L-1 of DA. Supporting electrolyte:
NaCl 0.5 mol L-1, pH 4.0 and = 30 mV s-1.

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Blum, S. A.; Zahrebelnei, F.; Nagata, N.; Zucolotto, V.; Mattoso, L. H. C.; Pessoa, C. A.; Garcia, J. R.; Wohnrath, K.

Table I. Electrochemical parameters obtained from the cyclic voltammograms of GRPE, KP0PE,
CNTPE, KPyPE and KPPE electrodes in the presence of 50 µmol L-1 DA

CPE Epa (mV) Epc (mV) p

= Epa- Epc (mV) Ipa (µA) Ipc (µA) Ipa/Ipc

GRPE 530.0 304.0 3.88 -3.50 -1.11

CNTPE 380.0 297.0 83.0 4.25 -1.20

KP0PE 484.0 280.0 199.0 -2.70 -0.99

KPyPE 433.0 329.0 102.0 2.82 -2.70 -1.04

KPPE 454.0 387.0 5.98 -0.99

kinect for electron transfer for DA redox process are KPPE and KPyPE, even considering the presence
of a high capacitive current. This behavior can be attributed to the smaller values of Ep
respectively (criteria for reversible process: p
= 57/n mV at 25 0C n is the number of electrons
involved in the process and Ipa/Ipc =
KPy20 can be assigned to the presence of the pseudocapacitance phenomena that is caused by fast
faradaic processes occurring on the material surface as a consequence of the presence of polymeric
®

S3), in the same experimental conditions.

We observed that oxidation of DA on a Pt electrode occurred at higher potentials (Epa= 504 mV). Another

-1

(Figure S4

advantageous for quantitative determinations.

µmol L-1.

Table II.
Linear Range of
Linear Regression Linear correlation Sensitivity
CPE Concentration
Equation factor (A mol-1 L)
(mol L-1)
GRPE 0.9930 0.051 2 – 50

CNTPE 0.04 2 – 50

KP0PE 0.9997 0.043 2 – 50

KPyPE Ipa = 0.9983 0.021 2 – 50

KPPE Ipa = 0.9958 0.110 0.04 2 – 50

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containing Eeonomer®

is also important to mention that the higher sensibility of KPPE electrode is clearly indicated in Figure 3. It
is also interesting to notice that the error on the measurement of the peak current for each of the studied

peaks on the KPPE.

a b
Figure 3. Analytical curves obtained by cyclic voltammetry in presence of DA for a) KPPE electrode and
b) CNTPE electrode, both in the concentration range of 2 to 50 µmol L-1 of DA. Supporting electrolyte:
NaCl 0.5 mol L-1, pH 4.0 and = 30 mV s-1.

In addition to the good sensitivity, a suitable reproducibility and response stability is required for an

in the presence of DA (Figure S5

n1/2
vs plot exhibited the typical behavior of a typical
1/2

electrochemical-chemical catalytic process, as observed for the electrodes containing KP20 and KPy20.

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Experimental Design to Enhance Dopamine Electrochemical Detection Using Carbon Paste Electrodes

a b

Figure 4. Plot of Ip/ 1/2 against scan rate ( ) for DA (50 µmol L-1 a) KPPE and
b) KPyPE. Supporting electrolyte: NaCl 0.5 mol L-1 and pH 4.0.

Electrochemical oxidation of ascorbic acid as a possible interferent for DA detection

and its presence considerably decreases the sensitivity of most of the materials used in electrochemical

(KPPE,
®

µmol L AA. An irreversible oxidation

-1

ca.

is highlighted as an insert in Figure 5.a due to the high capacitive current presented by this material. For

in the presence of AA. For those carbonaceous materials, it is possible to observe the same oxidation

presence of DA.
Braz. J. Anal. Chem., 2021, 8 (32), pp 178–197.

a b

Figure 5. Cyclic voltammograms of a) KP0PE, KPPE and KPyPE and b) KP0PE, GRPE and CNTPE in
presence of AA in a concentration of 50 µmol L-1. Conditions: NaCl 0.5 mol L-1, pH 4.0 and = 30 mV s-1.

The obtained results indicate that the use of KP0PE, GRPE and mainly CNTPE electrodes are not

feature for applications in biological samples.

the application in the preparation of electrochemical sensors. Aiming to produce a proof of concept for the

sensitivity (0.11 A mol-1

µA) and DA concentration (µmol L-1

µmol L-1
labeled concentration of DA in the tested commercial injection solution sample (50.4 µmol L-1).

Figure 6. Ipa vs DA concentration plot obtained for the

conditions: supporting electrolyte: NaCl 0.5 mol L-1, pH 4.0

and = 30 mV s-1.

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Blum, S. A.; Zahrebelnei, F.; Nagata, N.; Zucolotto, V.; Mattoso, L. H. C.; Pessoa, C. A.; Garcia, J. R.; Wohnrath, K.

Factorial Design
®
could enhance the

this process, a 32

due to the importance of this parameter in doping the conducting polymers present in the system.
®
KP20 (KPPE), KPy20

higher than 20 µmol L-1. Thus, the Eeonomers® (KP20 and KPy20) are highly selective to DA even in the
presence of its principal interferent (AA) in physiological medium.
The current results obtained in each assay of the 32 full factorial are displayed in the Table III. A central

Table III. Current results obtained in each assay of the 32 full factorial design to investigate the

Assay pH CPE composition Ipa (µA)

1 - (3.0) - (KP20) 28.3

2 - (KP20) 15.2

3 - (3.0)

4 9.31

5 0 (5.0) 0 (KP20/KPy20) 24.3

0 (5.0) 0 (KP20/KPy20) 24.0

7 0 (5.0) 0 (KP20/KPy20) 25.4

8 0 (5.0) - (KP20) 25.1

9 0 (KP20/KPy20) 20.4

10 0 (5.0) 7.82

11 - (3.0) 0 (KP20/KPy20) 22.0

vs

µ
f
- c = -9.71 units, obtained
2
factorial and the mean at the center of the design)

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Table IV. Analysis of Variance for the quadratic adjusted model (Ipa = 24.2 – 2.00 pH – 7.47 EPC
- 2.44 pH2 – 7.18 EPC2 + 3.94 pH x EPC)
Sum of Squares Degrees of Mean of Squares
Source F-test
(SS) Freedom (MS)
Regression (R) 5
52.39
Residual (r) 11.51 5 2.31

Lack of Fit (LF) 10.43 3 3.48

Pure Error (PE) 1.09 2 0.54

Total 10

F = MSR/MSr = 52.39
>>> F FLF = MSLF/MSpe F

µA) can be
obtained using the pH close to 3.0 and KP20 as EPC. Regarding the determination of DA in biological
samples (pH µ

Figure 7. Response surface of the model (Ipa = 24.2 – 2.00 pH – 7.47 EPC -
2.44 pH2 – 7.18 EPC2

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Experimental Design to Enhance Dopamine Electrochemical Detection Using Carbon Paste Electrodes

CONCLUSIONS
Eonomers® materials, prepared by incorporation of conducting polymers (Pani (KP20) and PPy

sensitivity of 0.110 A mol -1

presented good response stability in the presence of DA. The good analytical features of KP20 CPE
enabled its successful application to detect DA in a commercial formulation.

important interfering species for DA detection in physiological matrices. Both Eonomers®-based CPE

The response surface obtained after performing the 32

that the developed electrochemical devices can found promising applications to perform fast, cheap and
sensitive determinations of DA in real samples.

Acknowledgements

Education Personnel (CAPES).

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SUPPORTING INFORMATION

S1 – Scanning Electron Microscopy (SEM) Morphological analysis of pristine Eeonomers® by SEM

morphology of composites aimed to dispersion analysis of Eeonomers® particles in the elastomeric matrix
Eeonomers
fractured composites.

a b

Figure S1. (a) Photomicrographs obtained by SEM from

the Eeonomer® KP0 (a); KP20 (b) and KPy (c). Increase at
c 70000x.

S2 – Eonix Co provides Eeonomers®

electrochemical signals of dopamine at the carbon black electrode in the presence and absence of ascorbic

faradaic currents related to the redox processes of chemical doping of polyaniline in the emerald state can

193
Experimental Design to Enhance Dopamine Electrochemical Detection Using Carbon Paste Electrodes

Figure S2. Cyclic voltammograms for CPE/KP3.5 and CPE/

KP40 at 30 mV s-1 scan rate. Supporting electrolyte: NaCl 0.5
mol L-1 at pH 1.0. Inset: cyclic voltammograms of CPE/KP20

S3 –The oxidation of DA on a Pt electrode occurred at

higher potentials (Epa = 504 mV) in relation to electrodes

disadvantage on the use of Pt electrode is that DA

Figure S3.
mmol.L-1 of DA. Supporting electrolyte: NaCl 0.5 mol L-1, pH
-1
.

S4 – We observed that oxidation of DA on a Pt electrode occurred at higher potentials (Epa = 504 mV).

a b
Figure S4. Cyclic voltammograms for the KP0PE (a), KPPE (b), KPyPE (c) in 50 mol L-1 of DA.
Supporting electrolyte: NaCl 0.5 mol L-1, pH 4.0 and = 10, 30, 50. 70 and 90 mV s-1.
194
Braz. J. Anal. Chem., 2021, 8 (32), pp 178–197.

Figure S4 continuation. Cyclic voltammograms for the

KP0PE (a), KPPE (b), KPyPE (c) in 50 mol L-1 of DA.
Supporting electrolyte: NaCl 0.5 mol L-1, pH 4.0 and = 10,
30,
c 30, 50, 70 and 90 mV s-1.

vs the number of cycles, are illustrated.

a b

Figure S5. Cyclic voltammograms for the KP0PE (a), KPPE

c (b), KPyPE (c) in 50 mol L-1 of DA. Supporting electrolyte:
NaCl 0.5 mol L-1, pH 4.0 and = 30 mV s-1.

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Blum, S. A.; Zahrebelnei, F.; Nagata, N.; Zucolotto, V.; Mattoso, L. H. C.; Pessoa, C. A.; Garcia, J. R.; Wohnrath, K.

electrochemical behavior of each one before the simultaneous addition of dopamine and ascorbic acid.
From the concentration of 1.0 mmol L-1, the KP20 electrode began to detect ascorbic acid, but at this

from the concentration of 0.1 mmol L-1 have already detected ascorbic acid.

CPE Ipa vs E Ipa vs E

GRPE

KP0PE

CNTPE

Figure S6.
(3.0 x 10-5 to 7.0 x 10-5 mol L-1 -5
mol L-1, the
concentrations of ascorbic acid varied from 1.0 x 10-5 mol L-1 to 7.0 x 10 3 mol L-1.
Braz. J. Anal. Chem., 2021, 8 (32), pp 178–197.

CPE Ipa vs E Ipa vs E

KPy20

KP20

Figure S6 continuation.
CNTPE in DA (3.0 x 10-5 to 7.0 x 10-5 mol L-1 -5

mol L-1, the concentrations of ascorbic acid varied from 1.0 x 10-5 mol L-1 to 7.0 x 10 3 mol L-1.

197