Carbon Nanotube Modified Screen Printed Electrode for

Dopamine Detection Based on Electrochemical Sensing

Robeth Viktoria Manurung1, a), Ahmad Rashid2, Yaya Sulaeman1 and Isa Anshori2

1
Research Center for Electronics & Telecommunication, Indonesian Institute of Sciences
Jl. Cisitu No.21/154D, Komplek LIPI Sangkuriang, Bandung, Jawa Barat, Indonesia
2
Bandung Institute of Technology
Jl. Ganesa No.10, Bandung, Jawa Barat, Indonesia
a)
Corresponding author: robeth.viktoria.manurung@lipi.go.id

Abstract. This study is reporting the thick film sensor or screen-printed carbon electrode (SPCE) modified by Carbon
Nanotube (CNT) for electrochemical sensing of dopamine (DA). The modification was carried out using the drop-casting
method. The carbon surface of the SPCE working electrode has been modified by several different materials such as
FMWCNT (with and without Nafion), MWCNT /AgNP, and MWCNT-DMF. Cyclic Voltammetry (CV) and Differential
Pulsed Voltammetry (DPV) testing on dopamine showed that FMWCNT (with Nafion), MWCNT /AgNP, and MWCNT-
DMF had a sensitivity of 56.9; 39.0 and 11.6 µAmM-1cm-2, the detection limit is 300, 220, and 8.1 nM, the quantification
limit is 1.01; 0.75, and 0.027 μM, and with an average measurement error of 0.29; 0.53, and 19.65, respectively. The CNT
modified SPCE has been successfully fabricated and applied as an electrochemical sensor for dopamine and shows good
sensitivity and selectivity.

INTRODUCTION
Dopamine or (2-aminoethyl) benzene-1,2-diol (DA) plays an essential role as neurotransmitters (neuromediators)
responsible for sending neural signals in human and animal bodies [1–5]. It is found in almost all brain tissue,
especially in the basal ganglia and substantia nigra; therefore, damage to brain tissue due to cerebral hypoxia can
affect extracellular dopamine content. The amount of dopamine concentration in the human body is significant for
learning, memory, human behavior, and cardiovascular and renal systems [1–3], [6–8]. Deficiency of dopamine can
cause Parkinson’s disease, schizophrenia, and other health problems such as drug addiction [9–12]. The average range
of dopamine concentration is from 0.1 to 0.4 nM in the blood and 0.1 to 2 μM in urine [13], [14]. Thus, in actual
samples, dopamine must be measured at relatively low concentrations and in the presence of various interferences, in
particular ascorbic acid (AA) and uric acid (UA). The measurement technique of dopamine and other catecholamines
which has used in clinics is liquid chromatography [15]. However, electrochemical methods of dopamine control
attract increasing attention because electrochemical sensing is promising because of a rapid, sensitive, selective, and
low-cost detection of various biomolecular analytes. Among electrochemical methods of dopamine sense, it was
reported on potentiometry with electrodes selective either to dopamine [16], [17]. or to ions involved in the dopamine
oxidation process, e.g IO4− [18]. Some data exhibited in reviews on several techniques of measurements of dopamine
show voltammetry as the predominant approach to measuring this analyte electrochemically [20-23]. Electrochemical
techniques are highly developed due to sensitive detection by simple procedures. Electrochemical methods can detect
dopamine because it has good electrochemical activity and is easily oxidized.
 One application of the electrochemical method is the Screen-Printed Carbon Electrode (SPCE). SPCE is an
electrochemical strip electrode that consists of a working electrode and a counter electrode based on carbon, and a
reference electrode. However, this carbon-based electrode is still less selective, so modifications are needed to increase
the electrode selectivity and sensitivity to dopamine. Much research has been given attention to developing
nanomaterials, which are used to enhance the performance of electrochemical sensors. Nanomaterials are commonly
used to take advantage of a large surface area to mobilize biomolecules. This usually increases the number of binding
sites available to detect a specific chemical analyte [24]. Electrochemical sensors use a variety of nanomaterials to
enhance performance. Carbon nanotubes (CNTs) are among the most interesting materials due to their unique
electronic, chemical, and mechanical properties [25]. CNT consists of SP2 carbon units with a diameter of several
nanometers and a length of several microns. CNT can be synthesized into two groups, multi-wall (MW) and single-
wall (SW), by electric arc discharge, laser evaporation, and chemical vapor deposition methods. CNT behaves like
metals or semicircles depending on the diameter and degree of helically. Due to the high electronic conductivity for
electron transfer reactions and better electrochemical and chemical stability in aqueous and non-aqueous solutions,
they are suitable for modifying various electrodes. Furthermore, it is very promising to create efficient electrochemical
sensors using electrodes modified by CNTS, as they stimulate electron transfer reactions in many small biologically
essential molecules and large biomolecules.
In this study, three types of materials will be used to modify the surface of the SPCE working electrode. Four types
of materials to be synthesized include FMWCNT (Functionalized Multi-wall carbon nanotube) (without and with
nafion), MWCNT/AgNPs, and MWCNT-DMF. Furthermore, electrochemical testing of the analyte can be carried out
using a potentiostat or galvanostat device by Cyclic Voltammetry (CV) and the analyte that will be used in this test is
a sample of dopamine.

THE MATERIAL AND METHODS

2.1 The materials and apparatus
The equipment used in this study consisted of: CorrTest CS350 potentiostat, Screen Printing De Haast, Screen
Maker 300 TT, ultrasonic cleaner, Scanning Electron Microscopy (SEM), substrate (alumina), oven, petri dish, hair
dryer, ceramic cutters, screens, beakers, analytical balance scales, commercial reference electrodes, measuring cups,
multimeters, power supplies, reflux tools and hoses, magnetic stirrer, magnetic stirrer bar, micropipette, and aluminum
foil, while the materials and reagents used consist of of CNT, carbon paste (C), silver paste (Ag), KCl, ulano 133,
distilled water, acetic acid, ulano 133 sensitizer, ulano 5 stencil remover paste, Ag/AgCl paste, alumina, acetone, ulano
line, thinner, paste dielectric composition, H2SO4, HNO3, aquades, formaldehyde, absolute ethanol, silver nitrate,
ammonia, dimethylformamide (DMF),nafion, phosphate buffered saline (PBS), dimethyl sulfoxide (DMSO),
dopamine (analyte).

2.2 Design and fabrication of SPCE

The design process is carried out using the CAD software. The initial material for the sensor fabrication was a 50
x 50 mm, 96% Al2O3 substrate. The structure consisted of four layers of Ag conductor for electrode pad, Ag/AgCl
reference electrode (RE), Carbon based working and counter electrode and dielectric passivation. Prior to screen
printing process, the design layout was transferred to stainless steel screens with 325 mesh size using standard photo
emulsion process. The first step in the sensor fabrication was formation of the electrode pad layer by direct screen
printing of Ag paste (ESL) on the Al2O3 substrate followed by oven drying at 150 oC for 15 min. Subsequently, the
resulting layer was then fired in a conveyor belt furnace for 45 min with peak temperature of 800 oC. The second step
was formation of carbon based working and counter electrode by direct screen printing of carbon paste and followed
by another oven drying at 150 oC for 15 min. Final step was formation of reference electrode by printing Ag|AgCl
paste and followed by oven drying at 150 oC. The resulting structure was then passivated using dielectric layer by
screen printing dielectric paste and followed by curing at 150 oC for 15 min.

2.3 SPCE surface activation

The SPCE surface activation process is carried out using potential cycling which can increase the functionality of
the carbon on the surface of the working electrode. The step to activate is to drop an acid solution (H2SO4 0.1 M) on
the surface of the SPCE electrode. The cyclic voltammetry (CV) test was carried out with a potential range between -
2.5 – 2.5 V. During the activation process, oxygen and hydrogen gas bubbles will form on the carbon surface.
2.4 Working electrode Modification
The modification of the working electrode aims to improve the performance of the SPCE. This process employs a
CNT solution that has been functionalized with a mixture of strong acids H2SO4 and HNO3 MWCNT/AgNP, and a
MWCNT-DMF. The modification process is carried out by drop 2-4 µL of solution onto the SPCE working electrode
followed by drying in the oven at 50 oC for 5 minute.

2.5 Carbon nanotube pre-treatment

Approximately 2 g of CNT was diluted in mixture solution of 60 ml H2SO4 and 20 ml HNO3. Then, this mixture
is stirred with a magnetic stirrer and heated by the reflux method for 3 hours at 80 oC. After that, the carboxylated
CNTs were filtered through Whatman 40 filter paper assisted by a vacuum pump. The filtration product was washed
with distilled water several times until reach pH of 5 then transferred to a petri dish and dried in a vacuum oven
overnight [26]. Finally, the carboxylated CNT chunks were grind into powder by a mortar for further process.

2.6 Synthesis of AgNPs coupled with MWCNT

A total of 0.5 g of carboxylated CNT was added to 25 ml of a mixture of 38% formaldehyde, absolute ethanol, and
DI Water with a volume ratio of 3:10:10. Then, 25 ml of a mixed solution of 3.5% silver nitrate and 25% ammonia
(1:2) was added slowly into mixed solution with vigorous stirring to maintain the pH range between 8-10 [27].

2.7 Synthesis MWCNT-DMF

1 mg of CNT was diluted with 1 mL of DMF. After that, the mixture was dissolved using a magnetic stirrer at
2000 rpm for 12 hours (Hamburg, Germany). After completion, it was continued with the sonication process for 1
hour using an ultrasonic bath. After the concentrated solution is ready, further dilution will be carried out by mixing
the solution with a mixture of DMF:H20 (1:1) to obtain a CNT solution with a final concentration of 0.5 mg.mL-1.

2.8 Sample Preparation

The sample preparation process for the characterization of CV and DPV was carried out using stock solutions of
dopamine in high concentrations of 5 mM and 0.5 mM. The solution was diluted using 10 mM PBS pH 7.4 to obtain
variations in concentration ranging from 0.1 – 3 M, 1 – 20 M, and 0.1 – 1.0 mM. The dopamine solution at each
concentration will be dripped onto the modified SPCE working electrode.

2.9 Electrochemical Measurement

Electrochemical measurements using cyclic voltammetric method were carried out to characterize the peak currents
of oxidation and reduction occur on the SPCE modified CNT working electrode while the dopamine was dropped. The
electrochemical measurement was conduct using dopamine samples for CNT-modified SPCE electrodes (FMWCNT
(with and without nafion), MWCNT/AgNP, and MWCNT-DMF).

THE RESULTS AND DISCUSSION

3.1 Design and Fabrication of SPCE
The result of design and fabrication pf SPCE can be seen on Figure 1. The design result will transfer into
transparency film pattern for thick film fabrication as shown on Figure 1a and SPCE prototype on Figure 1b.

Figure 1. The result of (a) design SPCE pattern, (b) prototype.

3.2 SPCE Surface Activation
The surface activation of SPCE has been needed to enhance electron transfer during the electrochemical process. The
activation has been conducted by drop H2SO4 solution on the surface of SPCE working electrode. The SPCE surface
activation characterization was carried out using ferrocyanide [Fe(CN)6]4+ 10 mM solution to monitor the oxidation and
reduction peaks which is produced. The voltammogram spectra before and after activation in a 10 mM ferrocyanide
solution is shown in Figure 2.

Figure 2. Voltammogram spectra of activated and un-activated SPCE using H2SO4.

3.3. Surface characterization

SEM (scanning electron microscopy) characterization was carried out to confirmed the morphology of the SPCE
working electrode which has been modified with CNT. The SEM image exhibited the reactivity of the electrode from
its surface structure and produced CNTs as shown in Figure 3a.

Figure 3. SEM image of (a) CNT by adding composition H2SO4:HNO3; 3:1, (b) AgNPs decorated on SPCE-MWCNT surface,
(c) MWCNT formed on SPCE surface.
AgNPs has been succeed to fabricated on the surface of MWCNT modified SPCE electrode by SEM image
confirmation on Figure 3b. Generally, the oxidized MWCNT has reduced the size with the average diameter range
from 100 – 150 nm (Aqel et.al). Interestingly, the SEM image on Figure 3c showed MWCNT has diameter around 35
– 80 nm.

3.4 Electrochemical Properties of CNT

The electrochemical properties of the three modifications of CNT (FMWCNT, MWCNT/AgNP, and MWCNT-
DMF) has been CV evaluation by detecting dopamine sample. There are 3 types of CV tests that will be carried out,
namely testing the performance comparison of bare SPCE and CNT modified SPCE working electrodes,
characterization SPCE electrode performance of dopamine concentration variation, and lastly, the influence of the
scan rate value.
3.4.1 The Characterization of SPCE Performance
The purpose of characterization is to observed the effect of SPCE modification with different materials can influence
the SPCE performance. 0.5 mM dopamine solution has been applied for the test and the scan rate is 50 mV/s with a
working voltage between -1 to 1 V. The CV measurement spectra of bare SPCE vs CNT modified SPCE in 0.5 mM
dopamine solution are shown in Figure 4.

Figure 4. The voltammogram spectra of bare SPCE vs CNT modified SPCE in 0.5 nM Dopamine solution.
The voltammogram spectra confirmed the CNT modification has been improve the oxidation peak current in 0.5
nM Dopamin in PBS buffer PH 7. The oxidation peak current of each SPCE can be measured as follow, bare SPCE,
FMWCNT (without nafion), FMWCNT (with nafion) , MWCNT/AgNP and MWCNT-DMF has oxidation peak
current 0,2165; 0,61408; 0,27304; 0,30233 and 0,32656 mA/cm2, respectively. Hence, MWCNT/AgNP modified
SPCE has superior oxidation peak current compare than another SPCE modification and bare SPCE as well. Based on
voltammogram spectra, it is clearly seen that FMWCNT (with nafion) has similar peak with MWCNT/AgNPs,
however, MWCNT/AgNPs has voltage separation gap between oxidation and reduction peak shorter than FMWCNT
(with Nafion). It is indicating that the electron transfer on MWCNT/AgNPs faster than other SPCE modification
including bare SPCE.

3.4.2 Evaluation SPCE performance due Dopamine

In this characterization, the dopamine solution has been diluted into several concentration range from 0.1 – 1.0
mM in PBS solution PH 7. The scan rate of CV is 50 mV/s with a working voltage range of -1 – 1 V. The
voltammogram spectra of the modified SPCE electrode due to dopamine concentration is shown in Figure 5.
Figure 5. Graph of oxidation peak current vs DA concentration for (a) MWCNT-DMF, (b) MWCNT/AgNPs, (c) MWCNT with
Nafion and (d) MWCNT without Nafion.

Based on the results of the CV measurements of the four SPCEs modification vs dopamine concentration range of
0.1 – 1.0 mM, it is seen that there is an increase in the current density value along with the increase in the concentration
of dopamine compounds. This increase in current density occurs because the electrons released in the working solution
during the oxidation process of dopamine compounds tend to be more as the concentration of dopamine compounds
increases. In addition, the resulting oxidation peak was seen to shift to a greater working stress as the dopamine
concentration increased. This increase in working voltage is caused by the greater energy required by the electrode to
oxidize dopamine compounds (DA) to dopamine-quinone (DQ) when there is an increase in the concentration of the
dopamine solution. The sensitivity DA sensor based on four modifications: MWCNT-DMF, MWCNT/AgNP, and
FMWCNT (with and without nafion) are 3.82; 5.6; 9.2 and 3.51 x 10-5 A/cm2, respectively.
The detection limit (LoD) of measurement is the smallest concentration that can generate a signal that can be
measured by the sensor. LoD is determined by the equation:
LoD =!"#$ (1)
$
The Sb is the standard deviation of the measurement from the measurement with the smallest analyte concentration,
in this study, the measurements has been conducted for 5-7 times in 10 mM PBS solution pH 7.4. Then, b is the
gradient of the linear regression plot curve. Based on this equation, the detection limit values of MWCNT/AgNP,
MWCNT-DMF, and FMWCNT (with nafion) were 0.30; 0.22 and 0.0081 mM, respectively.
The measurement error of repeated measurements in 0.5 mM dopamine sample for MWCNT/AgNP, MWCNT-
DMF, and FMWCNT (with nafion) are 0.29%, 0.53 %, and 19.63%, respectively. The data indicated that the
measurement error of MWCNT/AgNP and MWCNT-DMF, has produced very low percentage compare than
FMWCNT with Nafion because the stability MWCNT/AgNP, MWCNT-DMF are better than FMWCNT.
 CONCLUSION
In conclusion, a comprehensive study of electrochemical sensing platform for DA was developed based SPCE-
MWCNT/AgNPs, MWCNT-DMF, and FMWCNT (with and without nafion). Due to the high conductivity and
synergistic effects of MWCNT and AgNPs, the current response of DA is significantly improved. The detection limit
values of MWCNT/AgNP, MWCNT-DMF, and FMWCNT (with nafion) were 0.30; 0.22 and 0.0081 mM,
respectively. Additionally, among four modification which has been employed in SPCE, MWCNT/AgNP, MWCNT-
DMF showed good response for DA detection.

ACKNOWLEDGMENTS
This research was funded by National Research and Innovation Agency (BRIN/RistekDikti) under scheme
INSINAS Grant No. 12/INS/PPK/E4/2021. The authors thank the Research Center for Electronics and
Telecommunication, - Indonesian Institute of Sciences, Indonesia for supporting this work. The authors also
acknowledge the facilities, scientific and technical support from the Advanced Functional Materials (AFM)
Laboratory, ITB, and the Biotechnology Laboratory, Padjadjaran University, and all parties in Biomedical
Engineering Study Program, School of Electrical and Informatics Engineering, ITB, Bandung.

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