Chemical Physics Letters 739 (2020) 136969

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Chemical Physics Letters

journal homepage: www.elsevier.com/locate/cplett

Research paper

Carbon nanofiber modified with reduced graphite oxide for detection of T

ascorbic acid, dopamine, and uric acid
⁎
Krishna Prasad Aryala, Hae Kyung Jeonga,b,
a
Department of Physics, Institute of Basic Science, Daegu University, Gyeongsan 38453, Republic of Korea
b
Department of Materials-Energy Science and Engineering, Institute of Industry and Technique, Daegu University, Gyeongsan 38453, Republic of Korea

H I GH L IG H T S

• AExcellent
novel modified carbon nanofiber electrode as electrochemical sensor.
• Large potential
sensitivities and selectivity for ascorbic acid, dopamine, and uric acid.
• separation for ascorbic acid, dopamine, and uric acid.

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

Keywords: A sensitive and selective electrode material was prepared with carbon nanofiber (CNF), modified with reduced
Carbon nanofiber graphite oxide (RGO), for electrochemical detection of ascorbic acid (AA), dopamine (DA), and uric acid (UA).
Reduced graphite oxide The modified CNF-RGO electrode displayed sharp oxidation peaks for AA, DA, and UA at 0.06, 0.18 and 0.32 V,
Ascorbic acid respectively, with differential pulse voltammetry (DPV) and possessed large surface area and low impedance.
Dopamine
Sensitivities of 0.12, 1.19 and 0.14 µA/µM were obtained with DPV for various concentrations of AA, DA, and
Uric acid
Electrochemical detection
UA, respectively. Modified electrode also showed prominent peak to peak potential separation in mixture, de-
monstrating excellent selectivity.

1. Introduction cost, high sensitivity, wide selectivity, and excellent reproducibility

[6,7]. However, the detection of these biomolecules is a challenging
The concentration of biomolecules, which serves as indicator of task due to their overlapped oxidation potential when they are present
disease diagnosis, are should be monitored in living system according to in the same mixture. Better electroactive and selective electrochemical
their requirement [1]. Detection of small biomolecules such as ascorbic electrodes, therefore, are necessary and important to be developed for
acid (AA), dopamine (DA), and uric acid (UA) is an important task in biosensor applications [3,4].
analytical fields. AA is a water soluble vitamin and used for treatment Different carbon materials have been employed in the electro-
of diseases such as scurvy, mental illness, and cancer while the DA is analytical investigations because of chemical inertness and wide po-
also very important catecholamine neurotransmitter, playing sig- tential window [5]. Among them, carbon nanofiber (CNF) is an inter-
nificant role in the functions of the nervous system. Disorder in the level esting member of the carbon family, which has attracted much
of DA may result serious neurological problems. Similarly, UA is an- attention in the field of sensor application due to their unique structure,
other important biological molecule present in our body fluids and is electronic properties and relatively high mechanical properties. Struc-
one of the terminal products of purine metabolism. Abnormal levels of turally it has long, smooth, and circular structure, consists of many
UA can cause the several diseases such as gout, kidney diseases, hy- exposed edges at the tip of the circular column, and therefore is ex-
pertension, and cardiovascular [2–7]. pected to have higher sensitivity and adsorption capacity during the
High-performance liquid chromatography, capillary electrophor- electrochemical investigation of biomolecules [8,9]. Previously, the
esis, chemiluminescence, and electrochemical methods have been em- sensitivity of CNF had been further improved either by mixing with
ployed for the detection of biomolecules, and the electrochemical other reactive carbon derivative or by introducing other reactive het-
method is one of the promising and reliable methods because of its low eroatoms in it, resulting in improved electrochemical and

⁎
Corresponding author at: Department of Physics, Institute of Basic Science, Daegu University, Gyeongsan 38453, Republic of Korea.
E-mail address: outron@gmail.com (H.K. Jeong).

https://doi.org/10.1016/j.cplett.2019.136969
Received 8 October 2019; Received in revised form 5 November 2019; Accepted 15 November 2019
Available online 16 November 2019
0009-2614/ © 2019 Elsevier B.V. All rights reserved.
K.P. Aryal and H.K. Jeong Chemical Physics Letters 739 (2020) 136969

Fig. 1. (a) XRD results of GO and RGO, and SEM images of (b) RGO, (c) CNF, and (d) CNF-RGO.

electroanalytical performances of CNF for the detection of the biomo- temperature.

lecules [5,9]. The synthesis of reduced graphite oxide was confirmed by X-ray
Reduced graphite oxide (RGO) is an another reactive and modified diffraction spectroscopy (XRD, Cu Kα radiation, λ = 1.54 Å, D/Max-
derivatives of the carbon family. Therefore it also has been reported 2500/PC, Rigaku, Japan) and energy dispersive X-ray spectroscopy
extensively as improving and supporting material in the synthesis of (EDS, Ltd., S-4300, JEOL, and Japan). Surface morphology of the
electrochemical electrodes due to its high hydrophilic properties and modified electrodes is studied with scanning electron microscopy (SEM,
large surface area [10–15]. However, RGO has not been used to modify Ltd., S-4300, JEOL, and Japan). All electrochemical experiments are
CNF for the detection of AA, DA, and UA yet. It could be expected that carried out by using EC-Lab (Bio-logic, SP-150, France). Three-elec-
modified CNF with RGO, composites, will improve the surface area, trode cell was applied with the prepared modified CNF-RGO electrode,
wettability, and reactivity of CNF, resulting in better performance to- as a working electrode, the platinum wire as a counter electrode, and
wards the detection of AA, DA, and UA. silver/silver chloride (Ag/AgCl) as a reference electrode.
In the present work, CNF is modified with RGO, and the modified Electrochemical properties are observed by chronocoulometry, elec-
CNF-RGO electrode is used as the electrochemical sensor electrode for trochemical impedance spectroscopy, cyclic voltammetry, and differ-
the detection of AA, DA, and UA, separately as well as simultaneously in ential pulse voltammetry.
the mixture with cyclic voltammetry and differential pulse voltam-
metry. In addition, the electrochemical impedance, electrochemical
2.2. Preparation of the modified CNF-RGO electrode
active surface area, sensitivity, and peak potentials of the modified
electrodes are also investigated. Overall results show that the modified
First, graphite oxide (GO) was synthesized by the modified Brodie
electrode, CNF-RGO, possess significantly improved sensitivity and se-
method as described elsewhere [16]. 1 g of natural graphite (> 75%,
lectivity in the detection of AA, DA, and UA.
100 mesh, Sigma-Aldrich) was ground with 6.5 g of sodium chlorate
(99%, Sigma-Aldrich) and stirred in 20 mL of the nitric acid for 24 h.
2. Materials and methods GO powder was obtained after the vacuum filtration with deionized
water several times followed by drying in a vacuum oven at 60 °C for
2.1. Chemicals and apparatuses 24 h. Then, reduced graphite oxide (RGO) was synthesized as follows:
The GO powder was annealed at 500 °C for 1 h in argon atmosphere
Phosphate buffer saline, carbon nanofiber (CNF, powder, diameter with the ramping rate of 10 °C min−1. The obtained sample was named
of 100 µm, length of 20–200 µm), and potassium hexacyanoferrate (III) as RGO. At the same time, carbon nanofiber (CNF) was prepared by
(K3[Fe(CN)6]) were purchased from Sigma-Aldrich. Biomolecules such mixing 3 mg of CNF in 20 mL of isopropyl alcohol (IPA) followed by the
as ascorbic acid, dopamine, and uric acid were also purchased from magnetic stirring for 6 h at room temperature in order to increase
Sigma-Aldrich. All purchased chemicals were directly used without any dispersion. Then 6 mg of RGO was added into the dispersed CNF so-
further treatment, and all experiments were carried out at room lution to prepare the modified working electrode (CNF-RGO) with

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K.P. Aryal and H.K. Jeong Chemical Physics Letters 739 (2020) 136969

sequential magnetic stirring for 6 h followed by the filtration and concentration of K3[Fe(CN)6], D is diffusion coefficient of the K3[Fe
drying processes. Finally, CNF-RGO was obtained in powder form and (CN)6] (6.5 × 10−6 cm2/sec), and t is time (sec). Fig. 2b shows that the
used further for its characterization and electrochemical measurements. CNF-RGO electrode possessed the highest electrochemical active sur-
For the electrochemical measurement, 5 µL of the CNF-RGO solution, face area of 5.6x10-7 cm2, as expected, when compared with the CNF
prepared by mixing 5 mg of CNF-RGO into 3 mL of IPA solution, was and RGO electrodes. We can conclude that highest electrochemical
dropped into completely clean and dried surface of glassy carbon active surface area is obtained from three dimensional structure of CNF,
electrode. Similarly, two other samples, CNF and RGO, are also pre- which possess own surface area, and added RGO, which obviously in-
pared with the same concentration and processes to compare with the creased the surface area of electrode [5,19]. It also indicates that
modified CNF-RGO electrode. modified electrode, CNF-RGO, possess highest adsorption capacity,
which has key role to increase the accumulation and biomolecular in-
teraction on the surface. Therefore, the CNF-RGO electrode is expected
3. Results and discussion
to have enhanced interaction of AA, DA, and UA on its surface.

3.1. X-ray diffraction spectroscopy, surface morphology and surface area

3.2. Cyclic voltammetry

Firstly, we confirmed the synthesis of reduced graphite oxide (RGO)

Fig. 3a displays the cyclic voltammetry (CV) results of carbon na-
from graphite oxide (GO) by X-ray diffraction spectroscopy (XRD)
nofiber (CNF), reduced graphite oxide (RGO), and modified CNF-RGO
measurement. Fig. 1a displays that GO has typical intense peak (0 0 2)
electrode measured at the scan rate of 50 mV s−1 in the PBS solution. It
near at 15° while RGO possessed compressed and less intense peak at
shows that modified electrode, CNF-RGO, possessed highest current and
17° indicating shift of characteristic peak of GO after thermal treatment.
showed better electrochemical performance in compared to CNF and
To confirm it more clearly, we also plotted the normalized XRD peak for
RGO. Increased current on the surface of CNF-RGO should be due to
GO and RGO in between 0 and 1, which displays that RGO has very low
obtained higher conductivity and larger electrochemical active surface
intense and broad peak after thermal reduction. Presence of com-
area. Then all the electrodes were used again in presence of biomole-
pressed, less intense and broad peak after thermal treatment confirms
cules such as ascorbic acid (AA), dopamine (DA) and Uric acid (UA)
the synthesis of RGO successfully [16,17]. In addition to this, we also
using CV in phosphate buffer saline solution. With the addition of AA
analyzed the carbon and oxygen content of GO and RGO with energy
(Fig. 3b), larger irreversible oxidation peak was observed on the surface
dispersive X-ray spectroscopy (EDS) measurement and shown in
of CNF-RGO at the potential of 0.08 V as compared to the other two
Table 1, which also confirms the reduction of oxygen content in GO
electrodes CNF and RGO (Fig. 3b). And Fig. 3c shows the oxidation of
after thermal treatment.
DA in which CNF shows the broad peak around 0.37 V, while CNF-RGO
Morphology of RGO, carbon nanofiber (CNF), and CNF-RGO are
shows negatively shifted sharp peak at the potential around 0.21 V with
shown in Fig. 1. RGO has two dimensional surfaces with gaps between
improved reversibility, indicating rapid electron transfer on the surface
layers while CNF has one dimensional long and smooth structure. After
of the CNF-RGO electrode. Similarly, the CNF-RGO surface showed
mixing, RGO has appeared on the surface of CNF and between CNFs,
slightly negatively shifted sharp oxidation peak of UA at the potential
which shows rougher and more exposed edges with small sized flakes
around 0.3 V with slight improvement in the reversibility (Fig. 3d).
(Fig. 2c), therefore, expected to increase the adsorption of biomolecules
Furthermore, Fig. 3e shows selectivity of electrodes with the addi-
and biomolecular interaction followed by enhancing the sensitivity. In
tion of 0.2 mM of AA, 0.05 mM of DA, and 0.2 mM of UA into the PBS
addition, EDS result also confirmed the mixing of CNF and RGO
solution. CNF electrode showed broad oxidation peaks which seem to
showing 7 at% oxygen in CNF-RGO composite (Table 1). Therefore,
be very difficult to distinguish the AA, DA and UA peaks in the mixture
more reactive and large surface from both mixed CNF and RGO in
while the RGO showed the slightly better result than CNF but could not
modified electrode; compared to precursor CNF itself, with biomole-
be satisfied. However, CNF-RGO easily showed three distinguishable
cules is expected.
oxidation peaks of AA, DA, and UA. The potential differences between
The electrochemical impedance spectroscopy was measured in the
the oxidized three molecules were 0.16 V (AA and DA), 0.11 V (DA and
frequency range of 100 mHz to 100 kHz. The lowest impedance was
UA), and 0.27 V (AA and UA). All the CV results obtained in presence of
found in the CNF-RGO surface, and CNF showed the highest impedance
AA, DA and UA demonstrated that modified CNF-RGO electrode has the
(Fig. 2a), confirming that CNF-RGO synergistically improved their
highest electrochemical responses towards the detection of these mo-
conductivity. Increased conductivity should be due to nature of CNF,
lecules and which could increase the sensitivity. As we know that CNF
which possess more expose sites for easy migration of electrolytic ions
possess unique structures with highly exposed edges, more defect sites
and makes effective network with graphene material, and large surface
and significant electrical properties, therefore makes it easier to interact
area after mixing with RGO [18,19]. Furthermore, the electrochemical
with biomolecule [18,22]. On the other hand, RGO which also possess
active surface area was also measured by using chronocoulometry with
higher dispersion, excellent conductivity and larger electrochemical
probe ions (Fe2+/Fe3+) in the presence of 0.1 M K3[Fe(CN)6] as a
active surface area [14,15,23], therefore collectively increases the
mediator with Anson’s equation [10,20,21]:
performance of modified CNF-RGO electrode in the detection of AA, DA
Q = 2nFACD1/2 π −1/2t1/2, (1) and UA.

where Q represents the charge in coulombs, n is the number of electrons 3.3. Differential pulse voltammetry
participated in the redox reaction, F is Faraday’s constant (96,500 C/
mole), A is the electrochemical active surface area (cm2), C is Based on the obtained excellent properties such as higher con-
ductivity, larger electrochemical surface area of modified CNF-RGO
Table 1 electrode, differential pulse voltammetry (DPV) was used with different
EDS results of CNF, GO, RGO and CNF-RGO. concentration of biomolecules to investigate the sensitivity and linear
Elements CNF GO RGO CNF-RGO character of the modified CNF-RGO electrode. Fig. 4a shows the linear
relationship between current and concentrations of AA (50–350 µM) at
at% wt% at% wt% at% wt% at% wt% the surface of modified CNF-RGO electrode, due to oxidation of AA (at
around 0.06 V), resulting in the linear equation as Y = 0.12× − 4.85
C 97 96 67 60 80 75 93 91
O 3 4 33 40 20 25 7 9 (R2 = 0.98). Fig. 4b shows the oxidation of different concentrations of
DA at around 0.18 V and the linear relationship between the peak

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K.P. Aryal and H.K. Jeong Chemical Physics Letters 739 (2020) 136969

Fig. 2. (a) EIS and (b) Chronocoulometry results of CNF, RGO, and CNF-RGO.

Fig. 3. CV results of CNF, RGO, and CNF-RGO (a) without biomolecule, (b) with 0.5 mM of AA, (c) with 0.1 mM of DA, (d) with 0.2 mM of UA, and (e) with the
mixture consists of 0.2 mM of AA, 0.05 mM of DA, and 0.2 mM of UA.

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K.P. Aryal and H.K. Jeong Chemical Physics Letters 739 (2020) 136969

Fig. 4. DPV results and their calibration plots of CNF-RGO with (a) AA, (b) DA, and (c) UA followed by (d) simultaneous detection of 0.1 mM AA, 0.01 mM DA, and
0.1 mM UA.

current and concentrations of DA (10–60 µM) at the surface of modified area and reactivity of surface. And these obtained properties increased
electrode. The obtained linear equation was Y = 1.19× − 1.46 the biomolecular interaction on its surface and which attributed to
(R2 = 0.98). Similarly, linear character of the modified electrode was higher sensitivity and selectivity in the detection of DA, AA and UA
also obtained in the case of UA which was used in the concentration separately and simultaneously. Therefore, modified CNF-RGO electrode
range of 100–700 µM. The oxidation peaks of UA were found around could have wide scope of practical applications with the excellent se-
0.32 V and obtained linear equation was Y = 0.14× − 14.2 lectivity of AA, DA, and UA.
(R2 = 0.98), as shown in Fig. 4c. Then all the obtained sensitivities of
0.12 (µA/µM), 1.19 (µA/µM) and 0.14 (µA/µM) for AA, DA and UA are
compared with the results obtained by other electrochemical sensors 4. Conclusion
and summarized in Table 2 [2–5,15,24–30]. It indicates that obtained
sensitivities are significantly comparable and better than some of the In the present work, CNF was modified with RGO by the simple
previous electrochemical sensors used for the determination of AA, DA physical method, and the modified CNF-RGO electrode was in-
and UA. vestigated for the electrochemical sensor application. The modified
The modified electrode was also applied for the detection of AA, DA CNF-RGO electrode showed excellent sensitivity of 0.12 (µA/µM), 1.19
and UA simultaneously with DPV again, as shown in Fig. 4d. The CNF (µA/µM) and 0.14 (µA/µM) in the detection of DA, AA, and UA, re-
and RGO electrodes showed broad and indistinguishable peaks, in- spectively, as well as significant potential separation when used for
dicating that simultaneous determination of them is not possible on simultaneous detection. The excellent performances of modified elec-
their surface. However, the modified electrode showed three sharp and trode are attributed to large electrochemical active surface area and
distinguishable peaks at 0.03 V, 0.19 V and 0.32 V for AA, DA and UA, increased reactivity obtained from synergic effect of CNF and RGO. The
respectively. The modified electrode also shows the potential differ- synthesis of modified CNF-RGO electrode was very simple and easy, but
ences of 0.17 V (between AA and DA), 0.14 V (between DA and UA) and the sensitivity and selectivity of it were tremendous. It is, therefore,
0.29 V (between AA and UA). It can be seen that the potential se- expected that modified electrode could be applied for biosensor appli-
paration of 0.17 V (between AA and DA) is higher and comparable with cation.
some of the previously reported results [31–33] while the potential
separation of 0.14 V (between DA and UA) is also higher than many
previous reported results [11,34]. Declaration of Competing Interest
All results show that the modified CNF-RGO electrode could be used
for the detection of DA, AA and UA separately as well as for their si- The authors declare that they have no known competing financial
multaneous determination with significant potential separation. The interests or personal relationships that could have appeared to influ-
addition of RGO into CNF in modified CNF-RGO electrode improved ence the work reported in this paper.
both electrochemical properties such as electrochemical active surface

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K.P. Aryal and H.K. Jeong Chemical Physics Letters 739 (2020) 136969

Acknowledgements

This work
[29]

[36]

[37]
Ref.

[5]

[2]
This work was supported by Basic Science Research Program
through the National Research Foundation of Korea (NRF-
Sensitivity (µA/µM)

2016R1D1A3B04931018).

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