Journal of Molecular Liquids 168 (2012) 69–74

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Journal of Molecular Liquids

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

Sensitive voltammetric determination of epinephrine in the presence of

acetaminophen at a novel ionic liquid modified carbon nanotubes paste electrode
Toktam Tavana a, Mohammad A. Khalilzadeh b,⁎, Hassan Karimi-Maleh b,⁎, Ali A. Ensafi c,
Hadi Beitollahi d, Daryoush Zareyee a
a
Department of Chemistry, Qaemshahr Branch, Islamic Azad University, Qaemshahr, Iran
b
Department of Chemistry, Science and Research Branch, Islamic Azad University, Mazandaran, Iran
c
Department of Chemistry, Isfahan University of Technology, Isfahan 84156-83111, Iran
d
Environmet Department, Research Institute of Environmental Sciences, International Center for Science, High Technology & Environmental Sciences, Kerman, Iran

a r t i c l e i n f o a b s t r a c t

Article history: A novel ionic liquid modified carbon nanotubes paste electrode (IL/CNTPE) had been fabricated by using
Received 4 December 2011 hydrophilic ionic liquid 1-methyl-3-butylimidazolium bromide [MBIDZ]Br as a new binder. The IL/CNTPE
Received in revised form 6 January 2012 was characterized by a scanning electron microscope and voltammetry. Electrochemical behavior of epineph-
Accepted 13 January 2012
rine (EP) at the IL/CNTPE had been investigated in pH 7.0 phosphate buffer solution (PBS) by cyclic voltam-
Available online 30 January 2012
metry (CV), electrochemical impedance spectroscopy (EIS), chronoamperometry (CA) and differential pulse
Keywords:
voltammetry (DPV). The experimental results suggested that the modified electrode exhibited an electroca-
Epinephrine talytic activity toward the redox of EP. The electron transfer coefficient, diffusion coefficient, and charge
Acetaminophen transfer resistant (Rct) of EP at the modified electrode were calculated. The IL/CNTPE showed an excellent
Ionic liquid electrocatalytic activity for selective detection of EP in the presence of acetaminophen (AC) by using DPV.
Multiwall carbon nanotubes Detection limit of EP and AC was found to be 0.09 and 0.5 μM respectively. The proposed sensor was success-
Sensor fully applied for the determination of EP and AC in human urine, pharmaceutical, and serum samples.
© 2012 Elsevier B.V. All rights reserved.

1. Introduction
Catecholamine is a class of important compounds for message
transfer in the mammalian central nervous system. It is released by
the adrenal medulla in situations of psychological stress or low
blood sugar level [1,2]. Catecholamines are also used as drugs to
treat bronchial asthma, hypertension, myocardial infarction and car-
diac surgery [3]. EP, also known as adrenaline, one of the important
catecholamines, plays a central role during physical or mental stress
and also stimulates a series of actions of the sympathetic nervous sys-
tem (SNS) known as the “flight or fight response” [4]. The presence of
EP in the body affects the regulation of blood pressure and the heart
rate, lipolysis, immune system, and glycogen metabolism. It elevates
the blood sugar level by increasing catalysis of glycogen to glucose
in the liver, and at the same time begins the breakdown of lipids in
fat cells [5]. These important actions of EP also make it a potent dop-
ing agent and hence, it is also banned in competitive games by the
World Anti Doping Agency [6,7]. Low levels of EP have been found
⁎ Corresponding authors. Tel.: + 98 911 2540112 (Cell phone); fax: + 98 335
5452509.
E-mail addresses: khalilzadeh73@yahoo.com (M.A. Khalilzadeh),
h.karimi.maleh@gmail.com (H. Karimi-Maleh).
in patients with Parkinson's disease [8–10]. From the point of view
of medicine, it is a drug for emergency treatment in severe allergic
reaction, cardiac arrest and sepsis [11]. Therefore, the quantitative
determination of EP concentration in different human fluids, such as
plasma and urine, is important for developing nerve physiology, phar-
macological research and life science [12]. Numerous electrochemical
methods have been developed to determine EP due to its electroactive
nature [13–17]. Electroanalytical methods have attracted more atten-
tion in recent years for environmental and biological compounds deter-
mination due to their sensitivity, accuracy, lower cost, and simplicity
[18–49].
Acetaminophen (N-acetyl-p-aminophenol or Paracetamol) is a
long-established substance being one of the most extensively
employed drugs in the world. It is an antipyretic and analgesic drug
commonly used against mild to moderate pain or for reduction of
fever. It is also non-carcinogenic and an effective substitute for aspirin
for the patients who are sensitive to aspirin and safe up to therapeutic
doses. AC is metabolized predominantly in the liver where it gener-
ates toxic metabolites. The large scale therapeutic use of this drug
generated the need for the development of fast, simple and accurate
methodologies for the detection of AC; for quality control analysis
(in pharmaceutical formulations) and for medical control (in biologi-
cal fluids such as urine, blood and plasma) [50,51]. Several methods
have been used for the determination of AC in pharmaceutical formu-
lations and biological fluids including titrimetry [52], UV–vis

0167-7322/$ – see front matter © 2012 Elsevier B.V. All rights reserved.
doi:10.1016/j.molliq.2012.01.009
70 T. Tavana et al. / Journal of Molecular Liquids 168 (2012) 69–74
spectrophotometry [53], flow-injection [54,55] and chromatographic
methods [56,57]. Among these methods, electrochemical methods
maybe the most widely applied because of high sensitivity, simplicity
and reproducibility of this approach [58–61].
Recently room temperature ionic liquid [RTIL] has been used as a
new kind of modifier for a chemically modified electrode. RTIL is com-
posed entirely of ions and exists as a liquid at room temperature with
the characteristics of negligible vapor pressure and good solubility
and chemical stability. As a new green media, RTIL has many unique
electrochemical properties, such as higher ionic conductivity and
wider electrochemical windows [62–66].
Carbon nanotubes are kinds of porous nanostructure materials
which are currently in the forefront of materials research. Nanotubes
have the ability to mediate electron transfer reactions with electroac-
tive species in solution when used as an electrode surface modifier for
designing new electrochemical sensors [67–73].
In the present work, we describe the preparation of a new elec-
trode composed of multiwall carbon nanotubes/ionic liquid paste
electrode and investigate its performance for the voltammetric deter-
mination of EP in aqueous solutions. We also evaluate the analytical
performance of the modified electrode for simultaneous determina-
tion of EP and AC.
2. Experimental
2.1. Chemicals
All chemicals used were of analytical reagent grade purchased
from Merck (Darmstadt, Germany) unless otherwise stated. Doubly
distilled water was used throughout.
A 1.0 × 10 − 3 mol L − 1 EP solution was prepared daily by dissolving
0.0183 g EP (from Merck) in water and the solution was diluted to
100 mL with water in a 100-mL volumetric flask. The solution was
kept in a refrigerator at 4 °C in dark. More dilute solutions were pre-
pared by serial dilution with buffer solution.
A 1.0 × 10 − 3 mol L − 1 AC solution was prepared daily by dissol-
ving 0.0150 g AC (>98%) (from Fluka) in water and the solution
was diluted to 100 mL with water in a 100-mL volumetric flask. The
solution was kept in a refrigerator at 4 °C in dark. More dilute solu-
tions were prepared by serial dilution with water.
Phosphate buffer (sodium dihydrogen phosphate and disodium
monohydrogen phosphate plus sodium hydroxide, 0.1 mol L − 1) solu-
tions (PBS) with different pH values were used.
High viscosity paraffin (d = 0.88 kg L − 1) from Merck was used as
the pasting liquid for the preparation of the carbon paste electrodes.
Spectrally pure graphite powder (particle size b 50 μm) from Merck
and multiwall carbon nanotubes (>90% MWNT basis, d × l = (100 −
80 nm) × (5 − 9 μm) from Fluka were used as the substrate for the
preparation of the electrodes.
2.2. Apparatus
Cyclic voltammetry, impedance spectroscopy, and differential
pulse voltammetry were performed in an analytical system, Autolab
with PGSTAT 302N (Eco Chemie, the Netherlands). The system was
run on a PC using GPES and FRA 4.9 software. For impedance mea-
surements, a frequency range of 100 kHz to 1.0 Hz was employed.
The AC voltage amplitude used was 5 mV, and the equilibrium time
was 15 min. A conventional three-electrode cell assembly consisting
of a platinum wire as an auxiliary electrode and an Ag/AgCl/KClsat
electrode as a reference electrode was used. The working electrode
was either an unmodified carbon nanotubes paste electrode
(CNTPE), or an IL/CNTPE. The prepared electrodes with carbon nano-
tubes and without carbon nanotubes were characterized by scanning
electron microscopy (SEM). Infrared (IR) spectra were recorded with
a Shimadzu IR-460 spectrometer. 1H-nuclear magnetic resonance
(NMR) spectra were recorded on a Bruker DRX-500 AVANCE instru-
ment in acetone-d6 at 500.1 MHz; δ is reported in parts per million
(ppm) and J in Hz. Elemental (C, H, N) analysis was performed with
a Heraeus CHN-O-Rapid analyzer. The qualitative analysis of products
was conducted with a HP 6890/5973 GC–MS and quantitative analy-
sis was conducted with HP 6820 GC equipped with a FID detector.
2.3. Preparation of 1-methyl-3-butylimidazolium bromide
The mixture of the 1-methylimidazole (1.58 mL, 0.02 mol) and 1-
bromobutane (2.1 mL, 0.02 mol) was stirred at room temperature for
20 min and then was refluxed for 12 h in Toluene. The reaction mix-
ture was cooled to room temperature and the resultant viscous 1-
methyl-3-methylimidazolium bromide was dried in vacuum for 5 h
and washed with ether. The crude product was purified by crystalliza-
tion from acetonitrile to give compound 3 (0.33 g, 75.5%).
2.4. Spectral data synthesis of 1-methyl-3-butylimidazolium bromide
(C8H15N2Br) yield: 0.33 g (75.5%); IR (KBr): (cm − 1) 3173 (m),
31220 (m), 2981 (m), 2952 (m), 2894 (m), 1583 (s), 14752 (s),
1391 (m), 1355 (m), 1183 (s), 1041 (s), 773 (m) cm − 1. 1H NMR (ac-
etone-d6): δ = 0.90 (t, 3 H, CH2CH2CH2CH3, J(HH) = 7.2 Hz), 1.94 (m, 2
H, CH2CH2CH2CH3, 3 J(HH) = 7.2 Hz), 1.98 (m, 2 H, CH2CH2CH2CH3,
3
J(HH) = 7.2 Hz), 4.13 (s, 3 H, NCH3), 4.44 (t, 2 H, N CH2CH2CH2CH3,
3
J(HH) = 7.2 Hz), 7.98 (d, 2 H, H 4,5(Im)), 10.22 (t, 1 H, H 2 (Im)). Anal.
Calcd. for C8H15N2Br (219.14): C 43.50, H 6.83, Br 36.45. Found: C
43.12, H 6.62, Br 36.97.
2.5. Preparation of the modified electrode
To eliminate any metal oxide catalysts within the nanotubes, mul-
tiwall carbon nanotubes were refluxed in the 2.0 M HNO3 for 12 h,
and then washed with twice-distilled water and dried at room tem-
perature. CNTPE was prepared by hand-mixing of 0.900 g of graphite
powder and 0.100 g multiwall carbon nanotubes plus paraffin at a
ratio of 70/30 (w/w) and mixed well for 40 min until a uniformly wet-
ted paste was obtained. The paste was then packed into a glass tube.
Electrical contact was made by pushing a copper wire down the glass
tube into the back of the mixture. When necessary, a new surface was
obtained by pushing an excess of the paste out of the tube and polish-
ing it on a weighing paper. IL/CNTPE was prepared by mixing of 0.27 g
of 1-methyl-3-butylimidazolium bromide, 0.80 g of the liquid paraf-
fin, 0.20 g of multiwall carbon nanotubes, and 0.80 g of graphite pow-
der. Then the mixture was mixed well for 40 min until a uniformly
wetted paste was obtained. A portion of the paste was filled firmly
into one glass tube as described above to prepare IL/CNTPE.
2.6. Preparation of real samples
Injection solution was prepared (1.0 mg mL − 1, DarouPakhsh
Company, Iran) and then 1.0 mL of the solution plus 10 mL of
0.1 mol L − 1 buffers (pH 7.0) was used for the analysis.
Urine samples were stored in refrigerator immediately after col-
lection (from the Isfahan University Health Centre). Ten milliliters
of the sample was centrifuged for 20 min at 1500 rpm. The superna-
tant was filtered out using a 0.45 μm filter and then diluted 5 times
with the PBS (pH 7.0). The solution was transferred into the voltam-
metric cell to be analyzed without any further pretreatment. The
standard addition method was used for the determination of EP in
real samples.
2.7. Recommended procedure
IL/CNTPE was polished with a white and clean paper. To prepare a
blank solution, 10.0 mL of the buffer solution (PBS, pH 7.0) was
 T. Tavana et al. / Journal of Molecular Liquids 168 (2012) 69–74
transferred into an electrochemical cell. The initial and final poten-
tials were adjusted to 0.00 and 0.35 V vs. Ag/AgCl, respectively. DPV
was recorded with a pulse height and a pulse width of 100 mV and
7 mV, respectively to give the blank signal and labeled as Ipb. Then,
different amounts of EP solution were added to the cell, using a
micropipette, and the DPV was recorded again to get the analytical
signal (Ips). Calibration curve was constructed by plotting the catalyt-
ic peak current vs. the EP concentration.
3. Results and discussion
3.1. Optimization of MWCNTs and the ionic liquid ratio
To obtain the best condition in the preparation of modified elec-
trode, the ratio of CNTs to ionic liquid in IL/CNTPE was optimized. The
result showed that with increasing the amount of CNTs10% w/w and
the ionic liquid (IL) in ration of 13.0% (to prepare the modified electrode
is the presence of fix amount of EP), the oxidation peak current for EP
increased and then it's leveled off. Therefore, we selected these condi-
tions (CNTs 10.0% and IL 13.0%) for preparation of modified electrode.
3.2. SEM characterization
Typical SEM images of different electrodes were shown in Fig. 1. It
can be seen that on the surface of carbon paste electrode (CPE)
(Fig. 1A), the layer of irregularly flakes of graphite powder was pre-
sent and isolated with each other. After CNTs were added to carbon
paste, it can be seen that CNTPE was distributed on the surface of
the electrode with special three-dimensional structure (Fig. 1B), indi-
cating that the CNTs were successfully modified on the IL/CNTPE.
3.3. Voltammetric study
Since EP has a catechol moiety, we anticipated that the redox
response of EP would be pH dependent (see Scheme 1). In order to as-
certain this, the voltammetric response of EP was obtained in solutions
with varying pH from 3.0 to 9.0 (Fig. 2 inset) at a surface of IL/CNTPE.
Result shows that the formal potential (E0/) of the redox couple was
pH dependent, with a slope of −56.2 mV/pH unit at 25 °C which was
equal to the anticipated Nernstian value for a two-electron, two-proton
electrochemical reaction (see Scheme 1). It can be seen that the maxi-
mum value of the peak current was appeared at pH 7.0 (Fig. 2), so this
value was selected throughout the experiments.
The active surface area of the modified electrode was estimated
according to the slope of the IP versus ν 1/2 plot for a known concen-
tration of K4Fe(CN)6, based on the Randles–Sevcik equation:
5 3=2 1=2 1=2
Ip ¼ 2:69  10 n AD ν CO ð1Þ
Fig. 1. SEM images of CPE (A) and IL/CNTPE (B).
71
Scheme 1. Oxidation reaction of epinephrine.
where Ipa refers to the anodic peak current, n the electron transfer
number, A the surface area of the electrode, DR the diffusion coeffi-
cient, C0 the concentration of K4Fe(CN)6 and ν the scan rate. For
1.0 mmol L − 1 K4Fe(CN)6 in 0.10 mol L − 1 KCl electrolyte with n = 1
and DR = 7.6 × 10 − 6 cm s − 1 and from the slope of the Ipa − ν 1/2 rela-
tion, the microscopic areas were calculated. They were 0.81, 0.071,
0.046 and 0.03 cm 2 for IL/CNTPE, CNTPE, IL/CPE and CPE, respectively.
The results show that the presence of CNTs and IL together causes the
active surface of the electrode to increase.
The direct electrochemistry of EP on the modified electrode was
investigated by cyclic voltammetry. Fig. 3 showed typical cyclic vol-
tammograms of different electrodes in a buffer solution (pH 7.0)
with a scan rate of 50 mV s − 1. For the bare carbon paste electrode
(CPE), an electrochemical signal of EP was obtained with the oxida-
tion peak current (Ipa) of 16.7 μA and the oxidation potential (Epa)
of 375 mV (Fig. 3, curve a). On the other hand, at CNTPE, the oxidation
peak current (Ipa) was obtained at 20.7 μA and the oxidation potential
(Epa) appeared at 355 mV (Fig. 3, curve b). These small changes in the
peak potential indicated that CNTs showed a little catalytic activity to
the EP oxidation. In addition, on the surface of bare IL/CPE, the oxida-
tion peak appeared at 315 mV with the peak current at 28.3 μA (Fig. 3,
curve c), which indicated that the presence of ionic liquids (ILs) in the
CPE could enhance the peak currents and decrease the oxidation
potential (decreasing the overpotential). The advantages of IL/CPE
had been elucidated with higher conductivity, fast electron transfer
rate, good anti-fouling properties and inherent catalytic ability of
ILs. So the oxidation peak current increased with decreasing of the
overpotential at IL/CPE, while on the IL/CNTPE, the oxidation peak
current increased to 33.8 μA with the oxidation peak potential as
310 mV (Fig. 3, curve d). The results indicated that the presence of
CNTs on IL surface had great improvement on the electrochemical
response, which was partly due to excellent characteristics of CNTs
such as good electrical conductivity, high chemical stability and
high surface area.
72 T. Tavana et al. / Journal of Molecular Liquids 168 (2012) 69–74
Fig. 2. Current–pH curve for electrooxidation of 350 μM EP at IL/CNTPE with a scan rate
of 50 mV s− 1. Inset) influence of pH on cyclic voltammograms of EP at the surface of
the modified electrode, (1–7: pH 3.0, 4.0, 5.0, 6.0, 7.0, 8.0 and 9.0, respectively).
The influence of potential scan rate (ν) on Ip of 200 μM EP at the
IL/CNTPE was studied by CV at various sweep rates (Fig. 4 inset). As
shown in Fig. 4, the peak currents of EP grow with the increasing of
scan rates and there are good linear relationships between the peak
currents and ν 1/2 (Fig. 4b). The regression equation is Ipa = 2.8661–
0.6331 ν 1/2 (Ipa: μA, ν: mVs − 1, R 2 = 0.9915), indicating the redox
process of 200 μM EP at the IL/CNTPE was diffusion-controlled. To ob-
tain further information on the rate determining step, a Tafel plot was
developed for the EP at a surface of IL/CNTPE using the data derived
from the raising part of the current–voltage curve (Fig. 5). The slope
of the Tafel plot is equal to n(1 − α)F / 2.3RT which comes up to
9.6696 V decade − 1. We obtained α as 0.43
Chronoamperometric measurements of EP at IL/CNTPE were car-
ried out by setting the working electrode potential at 300 mV vs.
Ag/AgCl/KClsat for the various concentrations of EP in buffered aque-
ous solutions (pH 7.0) (Fig. 6a). For an electroactive material (EP in
this case) with a diffusion coefficient of D, the current observed for
the electrochemical reaction at the mass transport limited condition
is described by the Cottrell equation [74]. Experimental plots of I vs.
t − 1/2 were employed, with the best fits for different concentrations
of EP (Fig. 6b). The slopes of the resulting straight lines were then
plotted vs. EP concentration. From the resulting slope and Cottrell
equation the mean value of the D was found to be 5.9 × 10 − 5 cm 2/s.
3.4. Electrochemical impedance spectroscopic study
Electrochemical impedance spectroscopy (EIS) is a valuable
method to monitor the impedance changes of the electrode surface
Fig. 3. Cyclic voltammograms of a) CPE, b) CNTPE, c) IL/CPE and d) IL/CNTPE in the
presence of 300 μM EP at pH 7.0, respectively.
Fig. 4. Plot of Ipa versus ν1/2 for the oxidation of EP at IL/CNTPE. Inset shows cyclic vol-
tammograms of EP at IL/CNTPE at different scan rates of 5, 10, 30, 50, 100, 150, 200 and
300 mV s− 1 in 0.1 M phosphate buffer, pH 7.0.
during the modification process. The semicircle diameters of Nyquist
plot reflect the electron transfer resistance (Rct), which is from the
electron transfer of the EP solution. Fig. 7 showed the EIS of differ-
ent modified electrodes in the presence of 400 μM EP. On the tradi-
tional CPE the value of Rct was obtained at 10.5 kΩ (curve a), which
was due to the presence of non-conductive liquid paraffin in the
carbon paste. After CNTs were added into the carbon paste to get
a CNTPE, the value of Rct was decreased to 8.5 kΩ (curve b),
which was smaller than that of CPE and was due to the presence
of conductive CNTs in the carbon paste. In continuation, with the
addition of IL in to CPE to get an IL/CPE, the value of Rct was
decreased to 6.2 kΩ (curve c), which was much smaller than that of
CPE and was due to the presence of high conductive IL in the carbon
paste. While on the IL/CNTPE (curve d), the value of Rct is equal to
5.18 kΩ, which is relative to the presence CNTs on the surface of IL/
CNTPE. All these results indicated that EP can successfully oxidize on
the surface of IL/CNTPE. It is interesting to note that the results
obtained (ratio of increase in oxidation current and decrease in Rct)
are very similar to those obtained from EIS and cyclic voltammetry.
3.5. Stability and reproducibility
The repeatability and stability of IL/CNTPE were investigated by
cyclic voltammetric measurements of 5.0 μmol L − 1 EP. The relative
standard deviation (RSD%) for ten successive assays was 0.92%.
Fig. 5. Tafel plot for IL/CNTPE in 0.1 M PBS (pH 7.0) with a scan rate of 10 mV s− 1 in the
presence of 200 μM EP.
 T. Tavana et al. / Journal of Molecular Liquids 168 (2012) 69–74
Fig. 6. Chronoamperograms obtained at IL/CNTPE in the presence of a) 100; b) 200;
c) 300 and d) 400 μM EP in the buffer solution (pH 7.0). Inset: Cottrell's plot for the
data from the chronoamperograms.
When using four different electrodes, the RSD% for seven measure-
ments was 1.2%. When the electrode is stored in the laboratory, the
modified electrode retains 98% of its initial response after a week
and 95% after 35 days. These results indicate that IL/CNTPE has good
stability and reproducibility, and could be used for EP.
4. Analytical features
DPV was used to determine EP and AC concentrations. The DP vol-
tammograms clearly show that the plot of peak current versus EP con-
centration is linear for 0.3–450 μM of EP, the regression equation being
Ip(μA)= (0.0237± 0.0020)CEP +(1.7250±0.1210) (r2 =0.9952, n =9)
while the regression equation for AC in the range of 1.0–600 μmol L− 1
is Ip(μA)= (0.01670± 0.0022)CAC + (1.1235± 0.0095) (r2 =0.9935,
n=8), where C is μM concentration of EP or AC and Ip is the peak current.
The detection limit was determined at 0.09 μM EP and 0.5 μM AC accord-
ing to the definition of YLOD =YB +3σ.
The main objective of this study was to detect EP and AC simulta-
neously using IL/CNTPE. Analytical experiments were carried out by
varying either the EP concentration in the presence of 500 μM AC in
a 0.1 mol L − 1 phosphate buffer (pH 7.0) using the modified electrode
(Fig. 8). It can be noted that the responses to EP at IL/CNTPE were rel-
atively independent of AC responses. The differential pulse voltam-
mograms showed two well-defined anodic peaks with a 200 mV
separation of the peaks (Fig. 8). Fig. 8 inset shows the dependence
Fig. 7. Nyquist plots of CPE (a), CNTPE (b), IL/CPE (c), and IL/CNTPE (d) in the presence
of 400 μM EP at the optimum condition.
73
of differential pulse voltammetric peak current on the concentration
of EP. Current sensitivities toward EP in the absence and in the pres-
ence of AC were found to be 0.0237 ± 0.0020 μA/μM (in the absence of
AC) and 0.0235 ± 0.0041 μA/μM (in its presence). It is interesting to
note that the sensitivities of the modified electrode towards EP in
the absence and presence of AC were virtually the same, which indi-
cates that the oxidation processes of EP and AC at the modified elec-
trode are independent and that simultaneous or independent
measurements of the two compounds are, therefore, possible without
any interference.
5. Interference studies
The influence of various substances as potentially interfering com-
pounds with the determination of EP was studied under the optimum
conditions with 1.0 μM EP at pH 7.0. The potential interfering sub-
stances were chosen from the group of substances commonly found
with EP in pharmaceuticals and/or in biological fluids. The tolerance
limit was defined as the maximum concentration of the interfering
substance that caused an error of less than ±5% for the determination
of EP. After the experiments, we found that neither 1000-fold of glu-
cose, sucrose, lactose, fructose, methanol and ethanol nor 800-fold of
Ca 2 +, Mg 2 +, SO42 −, Al 3 +, NH4+, F −, Na + and ClO4−, nor 500-fold
methionine, alanine, phenylalanine, valine, tryptophan and glycine
affected the selectivity. Nor did saturation of starch solution and
200-fold of urea and thiourea interfere with the determination of EP.
6. Real sample analysis
In order to evaluate the analytical applicability of the proposed meth-
od, also it was applied to the determination of EP and AC in injection
solution, serum, tablet and urine samples (see Table 1). To check the ac-
curacy of the proposed method for analysis of EP and AC and also com-
paring the results, published methods [16,26] were used to check the
obtained results. Those results clearly demonstrate and confirm the ca-
pability of the IL/CNTPE in the voltammetric determination of EP and
AC with high selectivity, accuracy, and good reproducibility.
7. Conclusion
An IL/CNTPE, using hydrophilic [MBIDZ]Br as a modifier, had been
fabricated and characterized by SEM and voltammetry. The electro-
chemical behavior of EP at the modified electrode was investigated
in pH 7.0 phosphate buffer solution. Compared with its response at
Fig. 8. DPVs of IL/CNTPE in 0.1 M phosphate buffer solution (pH 7.0) containing
575.0 μM AC and different concentrations of EP, from inner to outer: 90.0, 180.0,
240.0, 260.0 and 350.0 μM. Inset: Plot of the electrocatalytic peak current as a function
of EP concentration.
74 T. Tavana et al. / Journal of Molecular Liquids 168 (2012) 69–74

Table 1
Determination of EP and AC in real samples.

Sample EP added (μmol L) AC added (μmol L− 1) EP founded (μmol L− 1) AC founded (μmol L− 1)

Propose method (%) Recovery Published method [16] Propose method Recovery (%) Published methods [26]

Ampoule 5.0 – 5.2 ± 0.3 104.0 5.4 ± 0.5 – – –

15.0 – 15.3 ± 0.4 102.0 15.4 ± 0.6 – – –
a
Tablet – 20.0 – – – 19.8 ± 0.2 99.0 20.4 ± 0.5
– 30.0 – – – 30.5 ± 0.6 101.6 30.7 ± 0.8
Urine – – b LOD – b LOD b LOD – b LOD
10.0 15.0 10.3 ± 0.4 103.0 10.5 ± 0.6 15.5 ± 0.7 103.3 15.8 ± 0.9
20.0 20.0 20.3 ± 0.3 101.5 20.5 ± 0.7 19.7 ± 0.5 98.5 20.7 ± 1.0
Serum 40.0 50.0 40.7 ± 0.7 101.7 41.1 ± 1.1 49.6 ± 0.5 99.2 49.1 ± 1.0
a
300 mg tablet, Darou Pakhsh Company, Iran.

CPE, the electrochemical sensitivity of EP at the proposed electrode
was improved dramatically, revealing some advantages of IL/CNTPE
over CPE such as high conductivity and fast electron transfer. The
modified electrode successfully resolves the overlapped voltammetric
peaks of EP and AC by ≈ 200 mV, so that the modified electrode dis-
plays high selectivity in the DPV measurement of EP and AC in their
mixture solutions. The electrode was successfully used for the deter-
mination of EP and AC in injection solution, tablet, serum and urine
samples. In addition, the modified electrode exhibited a distinct
advantage of simple preparation, surface renewal, good reproducibil-
ity and good stability.
Acknowledgements
The authors wish to thank Qaemshahr and Science and Research
Branch, Mazandaran, Islamic Azad University, for their support.
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