Journal of Electroanalytical Chemistry 801 (2017) 503–510

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Journal of Electroanalytical Chemistry

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Electrochemical sensor for simultaneous determination of epinephrine and MARK

norepinephrine based on cetyltrimethylammonium bromide assisted SnO2
nanoparticles
N. Lavanya, C. Sekar⁎
Department of Bioelectronics and Biosensors, Alagappa University, Karaikudi 630003, Tamilnadu, India

A R T I C L E I N F O A B S T R A C T

Keywords: A highly sensitive and selective electrochemical sensor has been developed for simultaneous determination of
Electrochemical sensor catecholamine neurotransmitters, epinephrine (EP) and norepinephrine (NEP) using cetyltrimethylammonium
Neurotransmitters bromide (CTAB) assisted SnO2 nanoparticles (NPs) synthesized by microwave irradiation method. The products
Epinephrine were characterized by powder XRD, HRTEM, UV-visible absorption and FTIR spectroscopy and cyclic voltam-
Norepinephrine
metric methods. The CTAB-SnO2 NPs modified glassy carbon electrode (GCE) exhibited two sharp well defined
Surfactant
reduction peaks for EP and NEP with a significant potential difference of 0.354 V in phosphate buffer solution
SnO2 nanoparticles
(pH 5.0). Under optimum conditions, the calibration curves for EP and NEP were obtained over a wide con-
centration range of 0.1–250 and 0.1–300 μM with the lowest detection limits of 10 and 6 nM for EP and NEP
respectively. Interference studies showed that the modified electrodes have excellent selectivity towards EP and
NEP in the presence of potential biological interferents uric acid and ascorbic acid. The proposed electrode has
been successfully applied to the simultaneous determination of EP and NEP in human urine samples with sa-
tisfactory recoveries.

1. Introduction
Neurotransmitters control the physiological and behavioural con-
ditions in human beings by regulating communication within the neural
network. These molecules are involved in a large variety of neuro-
physiological processes including learning, sleep, memory and appetite.
Damage in secretion or uptake of neurotransmitters is known to cause
neurodegenerative diseases, drug addiction, and depressive syndromes
[1]. Among various neurotransmitters, two important catecholamines
epinephrine (EP) and norepinephrine (NEP) play essential roles in the
mammalian central nervous system (CNS). Excess of EP in the body
affects the regulation of blood pressure and heart rate, lipolysis, im-
mune system and glycogen metabolism. On the other hand, deficiency
of EP leads to Parkinson's disease. The EP also serves as a chemical
mediator for conveying the nerve pulse to different organs. When EP is
secreted into the bloodstream, it rapidly prepares the body for action in
an emergency. Hence the EP is used as a common healthcare medicine
for emergency treatment in severe allergic reaction, cardiac arrest and
sepsis [2,3].
NEP is also a significant transmitter which regulates blood pressure,
emotional arousal, and mood disorders. It promotes the conversion of
glycogen to glucose in the liver and helps in converting the fats into
fatty acids, resulting in an increment in energy production. The NEP is
responsible for the increased heart rate, dilation of pupils, dilation of air
passages in lungs and narrowing of blood vessels due to which body is
able to perform well in stressful situations [4]. Knowledge of the con-
centrations of plasma catecholamine and their metabolites is often
useful for diagnosis and evaluation of therapeutic and pharmacody-
namic effects for neurological, psychiatric and cardiovascular disorders.
Considerable efforts have been made to develop reliable methods for
the detection of EP and NEP which are structurally similar and often
coexist in biological samples. The identification and simultaneous de-
termination of these neurotransmitters using the electrooxidation pro-
cess become difficult due to their interference with each other during
identification. Therefore, the development of an efficient electro-
chemical sensor to monitor and determine catecholamines in real
samples is very important for non-invasive disease diagnosis and
pharmaceutical applications.
Various analytical methods including liquid chromatography [5],
chemiluminescence [6] and electrophoresis [7] have been employed for
the determination of catecholamines. These conventional methods are
time consuming, expensive and involve tedious procedures for sample

⁎
Corresponding author.
E-mail address: Sekar2025@alagappauniversity.ac.in (C. Sekar).

http://dx.doi.org/10.1016/j.jelechem.2017.08.018
Received 11 May 2017; Received in revised form 7 August 2017; Accepted 8 August 2017
Available online 09 August 2017
1572-6657/ © 2017 Elsevier B.V. All rights reserved.
N. Lavanya, C. Sekar
preparation. As a result, the determination of catecholamines by em-
ploying modified electrodes using distinct electrochemical techniques
has attracted more attention, because the approach does not require
pretreatment of samples and it may be performed in situ [8]. Individual
determination of EP or NEP has been reported by many researchers
[9–16]; however, the simultaneous determination of EP and NEP has
been rarely reported [8]. In many modified electrodes the determina-
tion of EP and NEP is impaired when ascorbic acid and uric acid are
present in the solution, causing overlapping of peaks. To the best of our
knowledge, there is only one report in the literature dealing with the
simultaneous detection of EP and NEP based on MWCNT modified edge
plane pyrolytic graphite electrode. In the present work, we report the
preparation of a new non-enzymatic electrochemical sensor based on
surfactant mediated SnO2 NPs to catalyze the reduction of EP and NEP
at well separated potentials. The fabricated sensor provides a platform
for a simple and sensitive voltammetric determination of the two
neurotransmitters EP and NEP simultaneously.
Tin dioxide (SnO2), an n type wide band gap semiconductor (3.6 eV
at 300 K), has attracted attention in various fields because of its unique
properties which are strongly dependent on the size, shape and or-
ientation [17]. Recently, SnO2 nanostructures have been used ex-
tensively for electrochemical sensing because of its high electro-
chemically accessible area, high electrical conductivity, high reactivity
and selectivity, as well as excellent chemical stability [18,19]. Nu-
merous methods have been developed to synthesize various nanos-
tructures that make more efficient use of the active surfaces by tuning
porosity, shapes, sizes, and facets which improve performance of the
sensors. In particular, surfactant mediated synthesis is extensively used
to decorate the chosen material for the purpose of enhancing both
physical and chemical properties. Herein, we have synthesized SnO2
NPs by microwave irradiation method using cationic cetyl-
trimethylammonium bromide (CTAB) and anionic ethylene diamine
tetra acetic acid (EDTA) as mediators and fabricated a novel electro-
chemical sensor for the selective and simultaneous determination of EP
and NEP by electrochemical reduction process. The attractive feature of
this method is the fact that the major biological interferents such as
ascorbic acid (AA) and uric acid (UA) do not undergo reduction and
hence their interference is greatly avoided.
2. Experimental
2.1. Materials
SnCl2·2H2O, centrimonium bromide (CTAB), ethylenediaminete-
traacetic acid (EDTA), Na2HPO4, NaH2PO4 and NH4OH were purchased
from Fischer Scientific, Mumbai. Epinephrine and norepinephrine were
bought from Sigma Aldrich. 0.1 M phosphate buffer solutions (PBS) of
various pH values were prepared by mixing the stock solutions of
Na2HPO4 and NaH2PO4 in appropriate ratios. All chemicals were of
analytical grade and were used without further purification and the
solutions were prepared using deionized water.
2.2. Surfactant mediated synthesis of SnO2 and fabrication of modified
electrodes
Surfactant mediated SnO2 nanoparticles were synthesized by mi-
crowave irradiation method. 0.1 M solution of SnCl2·2H2O was mixed
with 0.5 g (~ 20 wt% of SnO2) of CTAB under stirring at room tem-
perature [20]. Then, several drops of ammonium hydroxide (NH4OH)
were added to the above solution to reach a pH value of 8. The colloidal
solution was washed with deionized water and ethanol several times in
order to remove NH4+ and Cl− ions. The washed precipitate was ex-
posed to microwave irradiation at 600 W for 20 min. The final products
were annealed at 600 °C for 5 h in ambient atmosphere in order to
obtain SnO2 nanoparticles with improved crystallinity. The above
process was repeated for preparing EDTA assisted SnO2 and pristine
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Journal of Electroanalytical Chemistry 801 (2017) 503–510
SnO2 nanoparticles.
Glassy carbon electrodes (GCEs) were polished carefully with alu-
mina powder (1.0, 0.3 and 0.5 μm) on polishing cloth, rinsed with
deionized water and sonicated to remove the physically adsorbed
substances. SnO2 NPs (1.0 mg/1 mL) was dispersed in deionized water
by sonicating for about 20 min. Then, the prepared suspension (10 μL)
was dropped onto the GCE surface and dried at room temperature to
obtain SnO2 modified GCEs.
2.3. Instrumentation
Powder X-ray diffraction (XRD) patterns of the samples were re-
corded using a Bruker AXS D8 advanced diffractometer in the range of
20–80° with Cu Kα radiation (λ = 1.5406 Å). High resolution electron
microscopy (HRTEM), and selected area electron diffraction (SAED)
studies were carried out using a JEOL JEM 2100 microscope operated at
200 kV. Fourier transform infrared (FTIR) spectra were recorded an
AVATAR330 FTIR Thermo Nicolet spectrometer using the KBr pellet
technique. The optical spectra measurements were made using a UV–vis
spectrophotometer (Thermoscientific-Evolution 201).
Electrochemical measurements were performed with a CHI-900
electrochemical workstation (CH Instruments, USA) which consists of a
three electrode system comprising GCE (3 mm in diameter) as working
electrode, a platinum wire as auxiliary electrode and an Ag/AgCl (3 M
KCl) electrode as reference electrode. Square wave voltammetry (SWV)
measurements were performed in PBS (pH 5.0) containing EP and NEP
over the potential region from 0.6 to − 0.4 V at a frequency of 10 Hz,
amplitude of 50 mV and step potential of 5 mV.
3. Results and discussion
3.1. Structural analysis
Fig. 1A shows the powder XRD pattern of SnO2 powders synthesized
with and without mediators. The diffraction peaks could be indexed to
the tetragonal rutile structure of SnO2 (ICDD card No: 41-1445) with
space group P42/mnm. No additional peaks corresponding to possible
impurity phases such as CTAB and EDTA or other tin oxides were ob-
served in the XRD pattern up to the resolution limit of the instrument.
The average crystallite sizes, calculated using the Scherrer formula,
were found to be 13.09, 9.26 and 8.28 nm for pristine SnO2, EDTA and
CTAB assisted SnO2 respectively. It can be noticed that the presence of
mediators led to a moderate decrease in the peak intensity, accom-
panied by a pronounced broadening of the diffraction peaks.
Optical absorption spectra of surfactant assisted SnO2 samples
suggest that the absorption edge shifts to higher wavelengths implying
a red shift in the band gap with respect to bulk SnO2 (3.6 eV at 300 K).
The wavelength of absorption edge is observed at 483, 533 and 556 nm
for pristine SnO2, EDTA and CTAB assisted SnO2 (Fig. 1B) with band
gap energy (Eg) values of 2.95, 3.11 and 3.35 eV respectively (inset
Fig. 1B).
Fourier transform infrared spectroscopy (FT-IR) is usually employed
as an additional probe to evidence the presence of OH groups as well as
other organic and inorganic species. The FT-IR spectra of surfactant
assisted SnO2 nanoparticles are shown in Fig. 1C. In this spectrum, the
broad band at 3411 cm− 1 may be due to ᵞOH stretching vibration of
surface hydroxyl group or adsorbed water. The peak observed at
1636 cm− 1 is due to the stretching vibration of OH groups. Another
strong peak appeared in the low wave number region of 559 cm− 1
corresponding to the vibration of antisymmetric O-Sn-O mode of na-
nocrystalline SnO2 [21]. The observed increment in O-Sn-O band width
for surfactant assisted SnO2 NPs may be due to the decrement in crys-
tallinity, removal of residual organic impurities and increase of oxygen
content. The weak peaks observed at 2345 and 2940 cm− 1 for surfac-
tant assisted samples belong to the stretching vibrations of eCeH
bonds, which might originate from organic groups absorbed on the
N. Lavanya, C. Sekar Journal of Electroanalytical Chemistry 801 (2017) 503–510

Fig. 1. (A) XRD pattern, (B) UV–Vis absorption and (C) FTIR spectra of (a) pristine SnO2, (b) EDTA and (c) CTAB assisted SnO2 NPs.

Fig. 2. TEM (A & D), HRTEM (B & E) and

SEAD pattern (C & F) of pristine and CTAB-
SnO2 nanoparticles.

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N. Lavanya, C. Sekar
Fig. 3. SWVs obtained for the mixture of 500 μM each EP and NEP in 0.1 M PBS (pH 5.0)
at (a) bare GCE, (b) pristine SnO2, (c) CTAB-SnO2 (d) EDTA-SnO2 modified GCE.
surfaces of SnO2 nanoparticles.
Surface morphology of the pristine SnO2 and CTAB-SnO2 samples
were examined using TEM and HRTEM. As shown in Fig. 2A, pristine
SnO2 nanoparticles are mostly spherical in shape without any ag-
gregation. The average particle size is 15 nm, which is consistent with
the crystallite size calculated from the XRD patterns. The corresponding
HRTEM image clearly shows a single crystalline nature with the lattice
fringe width of 0.34 nm corresponding to a crystal plane of (110) and
Fig. 4. CVs of 500 μM (A) EP and (B) NEP in 0.1 M PBS (pH 5.0) at different scan rates (50–500 mV s− 1) at CTAB-SnO2/GCE; (C) & (D) corresponding plots of log υ vs. Ep/V.
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Journal of Electroanalytical Chemistry 801 (2017) 503–510
the surface is free from any lattice defects (Fig. 2C). However, the ad-
dition of CTAB led to the aggregation of SnO2 NPs (Fig. 2B) and surface
defects could be observed in the HRTEM image as shown in Fig. 2D. The
corresponding selected area electron diffraction (SAED) pattern
(Fig. 2E & F) confirms that the SnO2 NPs were of single crystalline
nature with lattice planes corresponding to (110), (101) and (200) of
SnO2 tetragonal phase. The results are in good agreement with the XRD
results.
3.2. Electrochemical behaviour of EP and NEP at CTAB-SnO2/GCE
The electrochemical behaviour of EP and NEP at different SnO2 NPs
modified glassy carbon electrodes was investigated by observing elec-
trocatalytic reduction in 0.1 M PBS (pH 5.0) by square wave voltam-
metry (SWV) method. Fig. 3 presents SWVs of EP and NEP at (a) bare
GCE, (b) SnO2/GCE, (c) CTAB-SnO2/GCE and (d) EDTA-SnO2/GCE. The
two neurotransmitters exhibited relatively poor current response at
bare GCE (curve a) and SnO2/GCE (curve b). Interestingly, when the
CTAB-SnO2/GCE (curve c) was used, the mixture displayed two well-
defined and sensitive peaks at − 0.045 V and 0.309 V corresponding to
the reduction of EP and NEP with a large peak separation of 0.354 V. It
was also noted that the cathodic peak currents were more than three-
fold higher (Ipc = − 39.9 μA for EP and −30.6 μA for NEP) when
compared to that of bare GCE and pristine SnO2/GCE. The enhanced
current response clearly indicates that the CTAB-SnO2 NPs accelerate
the rate of electron transfer with an excellent electrocatalytic activity
towards the reduction of EP and NEP. The significant potential se-
paration between EP and NEP (0.354 V) allows simultaneous
N. Lavanya, C. Sekar Journal of Electroanalytical Chemistry 801 (2017) 503–510

Scheme 1. Probable reaction mechanism for electro-reduction of EP and NEP at the CTAB-SnO2/GCE.

Fig. 5. SWVs obtained for various concentrations of (A) EP (0.1 to 250 μM) in the presence of 100 μM NEP and (B) NEP (0.1 to 300 μM) in the presence of 100 μM EP at CTAB-SnO2
modified GCE in 0.1 M PBS (pH 5.0) and (C) and (D) plots show the reduction peak current against the concentration of EP and NEP.

determination of EP and NEP in mixture samples without interference
from each other. Contrary to this, the electrochemical reduction of EP
and NEP is impaired in the anionic EDTA-SnO2 modified GCE (curve d),
which could be attributed to the oxygen enrichment during synthesis
and larger particle size when compared to the CTAB-SnO2 NPs Thus, the
cationic CTAB-SnO2 NPs have large surface area and large number of
defects when compared to that of pristine SnO2 and anionic EDTA-SnO2
NPs. The surfactant CTAB induced oxygen deficiency and higher sur-
face area of CTAB-SnO2 NPs promotes its electrocatalytic efficiency.
3.3. Effect of scan rate and pH
The influence of scan rate on the voltammetric response of EP and
NEP at CTAB-SnO2/GCE was studied over the potential range of − 0.4
to 0.8 V by CV. Fig. 4A–B shows the CVs of EP and NEP recorded at
different potential scan rates (υ) over the range of 50–500 mV s− 1 in
PBS (pH 5.0). The plots of peak current as a function of square root of
scan rate for two neurotransmitters are shown in the inset of Fig. 4. The
linear regression equations relating ipa with the scan rate over the range

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Table 1 Table 2
Comparison of different chemically modified electrodes for the determination of EP and Determination of EP and NEP in human urine sample using CTAB-SnO2/GCE.
NEP with CTAB-SnO2/GCE.
Sample EP concentration (μM) NEP concentration Recovery (n = 3) (%)
Electrode EP (μM) NEP (μM) Ref. (μM)

LR LOD LR LOD Added Found Added Found EP NEP

GR/Pd/GCE a
– – 5–500 0.064 9 1 1 0.9 1 1.2 98.5 102.4
MWNT/ZnO/CH/GCEb – – 0.5–30 0.2 10 2 5 4.8 5 5.3 99.2 98.5
DMSA/Au/GEc – – 0.1–700 0.055 11 3 10 9.8 10 10.1 97.4 98.7
Au/ITOd – – 0.1–25 0.087 12
PXSP/GCEe 2–390 0.1 – – 13
RuO2/CNT/HCF/GCEf 0.1–10 0.052 – – 14 RT (1 − α ) αF ∆Ep
SnO2/graphene/GCEh 0.2–200 0.017 – – 15 logks = α (1 − α ) + (1 − α ) α − log −
nFα 2.303RT
Nano-diamond/graphitei 0.01–10 0.003 – – 16
MWNT/EPPGEj 0.005–0.1 0.0015 0.005–0.1 0.0009 8 where n is the number on electrons involved in the reaction, ΔEp is the
CTAB-SnO2/GCE 0.1–250 0.01 0.1–1 0.006 This work
1–300 0.010
peak potential separation (Epa − Epc) and υ is the scan rate. The ks
value is calculated as 5.71 s− 1for EP and 5.66 s− 1 for NEP respectively.
a
Graphene palladium modified glassy carbon electrode. The pH of the supporting electrolyte (PBS) showed a significant
b
MWNTs-ZnO/chitosan modified glassy carbon electrode. effect on the electrochemical behaviour of EP and NEP at the surface of
c
Meso-2,3dimercaptosuccinic acid/Au-NPs modified gold electrode.
d
the modified electrode. SWVs were recorded for EP and NEP reduction
Au nanoparticles deposited indium tin oxide.
e
Poly p-xylenolsulfonephthalein modified glassy carbon electrode.
at CTAB-SnO2/GCE over the pH range 3.0–9.0. As shown in Fig. S1A,
f
Rutheniumoxide/hexacyanoferrate modified glassy carbon electrode. the peak currents of EP and NEP increased with increasing pH
h
SnO2/graphene modified glassy carbon electrode. value, reaching a maximum at pH 5.0, then decreasing with further
i
Pyrolytic graphite modified with nano-diamond modified graphite electrode. increase of solution pH. Subsequently, anodic peak potentials of EP
j
Multi walled carbon nanotubes modified edge plane pyrolytic graphite electrode.
and NEP shifted to less positive values as the solution pH increased.
The linear regression equations for EP and NEP were derived
of 50–500 mV s− 1, are found to yield: ipc (μA) = 3.86–1.55υ1/2
as Epc = 0.577–0.055pH (R2 = 0.998) and Epc = 0.6225–0.053pH
(mV s− 1) (R2 = 0.994) for EP and ipc (μA) = 1.97–1.11υ1/2 (V s− 1)
(R2 = 0.998) respectively (Fig. S1B). The plot of the peak potential (Ep)
(R2 = 0.999) for NEP respectively. The result suggests that both EP and
versus pH showed linearity with a slope of 55 mV/pH for EP and
NEP are diffusion controlled process. It was noted that the catalytic
53 mV/pH for NEP, which indicated that two electrons and two protons
oxidation peak potential (Epa) shifted to positive potentials with in-
take part in the electrode reactions (Scheme 1).
creasing scan rate for both species (Fig. 4C–D). The Epa versus the
logarithm of scan rate plots presented a linear relation, indicating that
the electrocatalytic oxidation of EP and NEP on the modified electrode 3.4. Individual and simultaneous determination of EP and NEP
surface is a typical quasi-reversible process. The linear regression
equation has been deduced as Epa (V) = 0.067log υ + 0.66 For the simultaneous determination of EP and NEP at the CTAB-
(R2 = 0.995) and Epc (V) = −0.059 log υ − 0.173 (R2 = 0.982) for EP SnO2/GCE, SWV was carried out in the potential range of 0.6 to − 0.4 V
and Epa (V) = 0.042 log υ + 0.55 (R2 = 0.9747) and Epc(V) with frequency of 10 s− 1 and pulse amplitude of 50 mV in PBS
= − 0.047log υ − 0.141 (R2 = 0.937) for NEP. According to Laviron's (pH = 5.0) because of its higher current sensitivity and better resolu-
equation [22], the plot of the peak potential vs. logarithm of scan rate tion than CV. Under identical conditions, the concentration of the target
yields two straight lines with slopes of −2.3RT / αnF and 2.3RT / (1-α) biomolecule was changed, while keeping the concentration of the other
nF for the cathodic and anodic peaks respectively. Electron transfer biomolecule as constant. Fig. 5A displays the SWVs obtained for dif-
coefficients (α) were calculated from the slopes as 0.5 and 0.62 for EP ferent concentrations of EP in the presence of 100 μM NEP at CTAB-
and NEP respectively. The apparent heterogeneous electron transfer SnO2/GCE. It was noted the cathodic peak current of EP increases lin-
rate constant (ks) can be obtained based on equation: early with its increasing concentration over a wide range of
0.1–250 μM, while the peak currents of NEP (100 μM) decreased

Fig. 6. (A) SWVs obtained for various concentrations of EP (1 to 300 μM) and NEP (1 to 250 μM) at CTAB-SnO2/GCE in 0.1 M PBS (pH 5.0) and inset shows plots of the reduction peak
currents as a function of various concentrations of EP and NEP and (B) Effect of interferences vs. recovery (%) recorded as SWV response of CTAB-SnO2/GCE for the addition of 100 μM
each EP and NEP in 0.1 M PBS (pH 5.0) with successive additions of 100-fold excess of interferents in the sequence of (a) NaCl, (b) KCl, (c) glucose, (d) serotonin, (e) thiamine, (f) uric
acid (g) ascorbic acid, (h) folic acid, and (i) dopamine.

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N. Lavanya, C. Sekar
slightly with the increase in the concentration of EP. This could be due
to the adsorption of reduced EP molecules on the surface of the mod-
ified electrode which becomes significant at higher concentrations. The
linear regression equation is expressed as Ipc = 0.0049x + 6.4
(R2 = 0.998) and the detection limit was calculated as 0.01 μM
(Fig. 5B). Similarly, voltammetric determination of NEP was carried out
in the presence of 100 μM EP. As shown in Fig. 5C, the peak current of
NEP increased linearly with increase in its concentration in the range of
0.1–1 and 1–300 μM with the linear regression equation (Ipc = 0.056x
+ 2.9 (R2 = 0.995)). The detection limit was calculated from the ca-
libration curve as 0.006 and 0.019 μM for EP and NEP respectively
(Fig. 5D). The results are compared with the values reported in the
literature for the determination of EP and NEP using various modified
electrodes (Table 1). It can be inferred from Table 1 that the perfor-
mance of the fabricated electrode (CTAB-SnO2/GCE) is superior when
compared to others in terms of its sensitivity, linear range and simpli-
city involved in the preparation methods. These results demonstrate
that the CTAB-SnO2/GCE could be a promising candidate for the si-
multaneous determination of EP and NEP in their binary mixture
without cross interference.
SWV responses of the sensor in the PBS (pH 5.0) containing
different concentrations of EP and NEP are shown in Fig. 6A. It can be
seen that the electrochemical reduction peak currents of EP and
NEP increased with increase in their concentrations. The linear re-
lationship exhibited between the peak currents and the concentrations
of EP and NEP in the ranges of 1–300 μM for EP, and 1–250 μM for
NEP with the detection limits of 0.014 and 0.013 μM (S / N = 3), re-
spectively. The linear relation could be expressed by regression equa-
tion as ipc = 0.092CEP − 7.9 (R2 = 0.997) and ipc = 0.048CNEP − 4.47
(R2 = 0.0997).
3.5. Interference, reproducibility and stability
The possible interferences of several ions and biomolecules on the
determination of EP and NEP were studied by SWV detection (Fig. 6B).
No interference was observed in the presence of Na, K, Mg (200-fold
concentrations), glucose, thiamine, uric acid, ascorbic acid, folic acid
and serotonin (10-fold concentrations). It is clear that the fabricated
sensor exhibits good selectivity for the determination of the two im-
portant neurotransmitters in the presence of large amount of several
potential interferents of physiological importance. Considering the fact
that ascorbic acid (AA) and uric acid (UA) are the main biological in-
terferents that can interfere with the voltammetric determination of EP
and NEP, a separate study was carried out to evaluate the effect of 100
fold excess of AA and UA for determination of epinephrine and nor-
epinephrine (Fig. S2). No reduction peaks were found for AA and UA
over the negative potential range studied, because of it being an irre-
versible process, indicating that these species do not interfere with the
simultaneous determination of EP and NEP. Therefore, the CTAB-SnO2/
GCE electrode can be used for determination of EP and NEP in real
samples using electrocatalytic reduction process.
To evaluate the reproducibility of CTAB-SnO2/GCE, the peak cur-
rents of SWV in binary mixture solutions containing 0.5 mM each EP
and NEP were recorded. The relative standard deviations (R.S.D.) of the
sensor in 6 successive measurements are found to be 2.7% and 3.2% for
EP and NEP respectively. Ten sensors prepared independently under
same conditions, revealed an acceptable repeatability with R.S.D. va-
lues of 4.1% and 3.2% for EP and NEP respectively. The stability of the
fabricated electrode was examined by storing it in air at room tem-
perature for two weeks. The peak current retained 98.7% for EP and
97.1% for NEP from its initial value. All these results indicate that the
CTAB-SnO2/GCE electrode has good reproducibility and stability.
3.6. Real sample analysis
The practicability of the newly developed CTAB-SnO2/GCE for real
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Journal of Electroanalytical Chemistry 801 (2017) 503–510
sample analysis was investigated by examining human urine samples by
the standard addition method. The urine samples were collected from
the Government Hospital, Karaikudi and were stored in a refrigerator
immediately after the collection. Different concentrations of EP and
NEP were added to the urine sample and the reduction peak currents
were measured by SWV method. The relative recovery of EP and NEP
for the spiked samples is in the range of 97.4% to 102.4%, and the RSD
(n = 5) is less than 2.0% (Table 2). The normal level of epinephrine and
norepinephrine in human urine is 0 to 109 nM and 0 to 473 nM re-
spectively. The developed CTAB-SnO2 sensor showed the lowest de-
tection limits of 10 and 6 nM for EP and NEP over wide dynamic ranges
of 0.1–250 and 0.1–300 μM respectively. Hence, these sensors could be
very well used for detection of EP and NEP in human urine and other
biological fluids successfully.
4. Conclusions
A novel electrochemical sensor has been developed for the si-
multaneous determination of EP and NEP based on CTAB-SnO2 mod-
ified glassy carbon electrode. The fabricated electrode exhibited ex-
cellent electrocatalytic activity towards the reduction of EP and NEP
with a large peak separation (0.35 V) which allows this sensor to detect
the target molecules either individually or simultaneously by the vol-
tammetric method. The modified electrode shows well defined cathodic
peaks for the neurotransmitters and they are not affected even when
coexisted at high concentrations. The CTAB-SnO2/GCE is a rapid,
simple, selective and sensitive protocol for simultaneous determination
of EP and NEP over a wide concentration range (0.1–250 μM for
EP & 0.1–300 μM for NEP) and low detection limit (10 nM for
EP & 6 nM for NEP). Furthermore, the developed electrode displays
acceptable non-interference ability with good reproducibility and sta-
bility. Therefore, it promises to serve as a useful tool for sensitive and
selective assays of EP and NEP in both research and clinical labora-
tories.
Acknowledgement
Authors NL & CS acknowledge the Department of Science and
Technology (DST-SERB –SB/S2/LOP-027/2013) for the financial as-
sistance.
Appendix A. Supplementary data
Supplementary data to this article can be found online at http://dx.
doi.org/10.1016/j.jelechem.2017.08.018.
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