Food Chemistry 210 (2016) 156–162

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Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Simultaneous determination of caffeine and paracetamol by square wave

voltammetry at poly(4-amino-3-hydroxynaphthalene sulfonic acid)-
modified glassy carbon electrode
Molla Tefera a,b,⇑, Alemnew Geto c, Merid Tessema a, Shimelis Admassie a
a
Department of Chemistry, Addis Ababa University, P. O. Box 1176, Addis Ababa, Ethiopia
b
Department of Chemistry, University of Gondar, P. O. Box 196, Gondar, Ethiopia
c
Department of Chemistry, Samara University, P. O. Box 132, Samara, Ethiopia

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

Article history: Poly(4-amino-3-hydroxynaphthalene sulfonic acid)-modified glassy carbon electrode (poly(AHNSA)/

Received 22 February 2015 GCE) was prepared for simultaneous determination of caffeine and paracetamol using square-wave
Received in revised form 22 March 2016 voltammetry. The method was used to study the effects of pH and scan rate on the voltammetric response
Accepted 21 April 2016
of caffeine and paracetamol. Linear calibration curves in the range of 10–125 lM were obtained for both
Available online 22 April 2016
caffeine and paracetamol in acetate buffer solution of pH 4.5 with a correlation coefficient of 0.9989 and
0.9986, respectively. The calculated detection limits (S/N = 3) were 0.79 lM for caffeine and 0.45 lM for
Keywords:
paracetamol. The effects of some interfering substances in the determination of caffeine and paracetamol
Poly(4-amino-3-hydroxynaphthalene
sulfonic acid)
were also studied and their interferences were found to be negligible which proved the selectivity of the
Square wave voltammetry modified electrode. The method was successfully applied for the quantitative determination of caffeine
Caffeine and paracetamol in Coca-Cola, Pepsi-Cola and tea samples.
Paracetamol Ó 2016 Elsevier Ltd. All rights reserved.
Coca-Cola
Pepsi-Cola and tea

1. Introduction migraine headache, backache and toothache (Kumary, Divya,

Caffeine (1,3,7-trimethylxanthine) is a naturally occurring alka-
loid belonging to N-methyl derivatives of xanthine, which is found
in tea leaves, coffee beans, cola nuts, cocoa beans and other plants.
It is used as a flavoring agent in a variety of beverages, including
some soft and energy drinks. Caffeine is also a central nervous
system stimulant. In moderate doses, it can increase alertness,
reduce fine motor coordination, cause insomnia, headaches,
nervousness and dizziness (Aly, 2013; Guo, Zhu, Yang, Wang, &
Ye, 2011; Habibi, Abazari, & Azar, 2012). However, intense use of
caffeine over time can lead to irritability, mutation effects such
as inhibition of DNA, anxiety and tremors, among other side effects
(Zhang et al., 2011). It can mobilize calcium from cells which leads
to bone mass loss and is considered as a risk factor for cardiovas-
cular diseases (Ali, Eisa, Taha, Abdalla, & Elbashir, 2012; Torres,
Barsan, & Brett, 2014).
Paracetamol (N-acetyl-p-aminophenol, acetaminophen) is an
effective and safe drug that is applied to reduce fever, relieve
coughing, colds and pain including muscular aches, chronic pain,
⇑ Corresponding author at: Department of Chemistry, Addis Ababa University,
P. O. Box 1176, Addis Ababa, Ethiopia.
E-mail address: mollatef2001@gmail.com (M. Tefera).
Nancy, & Sreevalsan, 2013; Shiroma, Santhiago, Gobbi, & Kubota,
2012; Tavallali & Hamid, 2011). In general, paracetamol does not
exhibit any harmful side effects but overdosing and chronic uses
produce toxic metabolite accumulation that will cause kidney
and liver damage (Kang et al., 2010). Paracetamol is often used in
the presence of other drugs like; aspirin, cetirizine, tramadol,
caffeine and codeine (An & Hoang, 2009; Shahrokhian & Saberi,
2011). A combination of caffeine and paracetamol is used in
migraine attack, child birth and avoid postpartum hemorrhage
(Chandra & Sharma, 2013). The use of the mixture of paracetamol
and caffeine as an analgesic and antipyretic is also well established
in pharmaceutical formulation (Vichare, Mujgond, Tambe, & Dhole,
2010). Therefore, it is very important to develop a simple, fast,
sensitive and accurate method for the detection of caffeine and
paracetamol in food, pharmaceutical products and biological
samples.
Different methods have been employed for the simultaneous
determination of caffeine and paracetamol which include spec-
trophotometry (Alpdogan, Karabina, & Sungur, 2002; Vichare
et al., 2010) and high-performance liquid chromatography (Dinc,
Yurtsever, & Onur, 2004; Tsvetkova et al., 2012). However, most
of the methods suffer from disadvantages such as long analysis
time, high running costs and requirement of sample pretreatment

http://dx.doi.org/10.1016/j.foodchem.2016.04.106
0308-8146/Ó 2016 Elsevier Ltd. All rights reserved.
 M. Tefera et al. / Food Chemistry 210 (2016) 156–162
which is time consuming, making them unsuitable for routine
analysis (Torres et al., 2014). As a result, electroanalytical method
is a method of choice for the simultaneous determination of
caffeine and paracetamol. Electrochemically both caffeine and
paracetamol were determined at the surface of self-assembled
monolayer of non-peripheral amine substituted copper(II)
phthalocyanine modified glassy carbon electrode (Jeevagan &
John, 2012), graphite-polyurethane screen printed electrode
(Saciloto, Cervini, & Cavalheiro, 2013), poly(taurine)/TiO2-
graphene nanocomposite modified glassy carbon electrode
(Xiong, Huang, Xu, Jin, & Zhai, 2013), boron-doped diamond
electrode (Lourenc, Medeiros, Filho, Mazoa, & Filho, 2009) and
Nafion/ruthenium oxide pyrochlore/GCE (Zen & Ting, 1997).
Furthermore, both caffeine and paracetamol were also determined
electrochemically in the presence of other drugs like aspirin
(Sanghavi & Srivastava, 2010) and noradrenaline and xanthine
(Aref, Raoof, & Ojani, 2014) and orphenadrine (Eisele, Clausen,
Tarley, Antonia, & Sartori, 2013).
Previous works in our laboratory reported the electrochemical
determination of caffeine at poly(4-amino-3-hydroxynaphthalene
sulfonic acid) modified glassy carbon electrode (poly(AHNSA)/
GCE) (Amare & Admassie, 2012) and paracetamol at poly(3,4-ethy
lenedioxythiophene) modified glassy carbon electrode (PEDOT/
GCE) (Mehretie, Admassie, Tessema, & Solomon, 2011). The results
showed that poly(4-amino-3-hydroxynaphthalene sulfonic acid)
modified glassy carbon electrode exhibited good sensitivity and
low detection limit (0.067 lM) for caffeine determination. A wider
linear range and high sensitivity with relatively high detection
limit (1.13 lM) were obtained for paracetamol at PEDOT/GCE. To
the best of our knowledge, there are no reports on the application
of poly(4-amino-3-hydroxynaphthalene sulfonic acid) modified
glassy carbon electrode for the determination of paracetamol and
the simultaneous determination of caffeine and paracetamol.
Hence, we report a simple and sensitive electrochemical method
for the simultaneous determination of caffeine and paracetamol
at poly(4-amino-3-hydroxynaphthalene sulfonic acid) modified
glassy carbon electrode.
2. Experimental
2.1. Reagents
Caffeine, paracetamol, 4-amino-3-hydroxynaphthalene sulfonic
acid (AHNSA), acetic acid and sodium acetate were purchased from
Sigma-Aldrich, Germany. Stock solutions of caffeine and paraceta-
mol were prepared freshly by dissolving a weighed amount in dou-
ble distilled water and then stored in refrigerator. A 0.1 M acetate
buffer solution in the pH range of 3.5–6.0 was employed as the
supporting electrolyte. Working standard solutions of both caffeine
and paracetamol were prepared by appropriate dilutions of the
stock solutions in acetate buffer just before use. All solutions were
prepared with double distilled water.
2.2. Apparatus
All electrochemical studies were carried out in a three
electrodes setup with glassy carbon electrode (GCE) or poly(4-am
ino-3-hydroxynaphthalene sulfonic acid) modified glassy carbon
electrode (poly(AHNSA)/GCE) as the working electrode, a platinum
wire as the counter electrode and a silver-silver chloride (in satu-
rated KCl) electrode as the reference electrode. A CHI760D Electro-
chemical Workstation, CHI Instruments (Austin, Texas, USA)
connected to a desktop PC was used to run all experiments. All
pH measurements were made with digital Jenway model 3345
ion meter.
157
2.3. Preparation of poly(4-amino-3-hydroxynaphthalene sulfonic acid)
modified glassy carbon electrode
Prior to electrode modification, the bare glassy carbon electrode
was polished with 1.0, 0.3 and 0.05 lm alumina powder and rinsed
with double distilled water. Finally, it was sonicated with doubly
distilled water in order to remove adsorbed particles, and then it
was allowed to dry in air. The electrochemical polymerization
process of 4-amino-3-hydroxynaphthalene sulfonic acid was
carried out by immersing the glassy carbon electrode in 0.1 M
HNO3 solution containing 2 mM 4-amino-3-hydroxynaphthalene
sulfonic acid monomer with a repetitive potential scan between

0.8 V and 2.0 V at a scan rate of 0.1 V s
1 for fifteen cycles. The
modified electrode was then carefully washed with double
distilled water and immersed in monomer free 0.5 M H2SO4
solution and the potential was scanned between 0.8 V and
0.8 V
(at a scan rate of 0.1 V s
1) until a stable voltammogram was
obtained.
2.4. Preparation of samples
Cola-beverages (Coca-Cola and Pepsi-Cola) were obtained from
a local supermarket, Addis Ababa and were used on diluting by a
factor of 1:10 (v/v) with acetate buffer after sonical elimination
of a gas. The tea solution was prepared by dissolving 15.0 g of
tea (Ethiopian Black Lion Tea) in 250 ml of double distilled water
followed by boiling for 1 h on a hot plate with stirring (Habibi
et al., 2012). The residue was allowed to settle and filtered through
a filter paper and diluted with acetate buffer by a factor of 1:10
(v/v). The dilution process helps to reduce the matrix effects. The
standard addition method was used for the analysis of Coca-Cola
and Pepsi-Cola and tea samples by spiking with aliquots of caffeine
and paracetamol.
3. Results and discussion
3.1. Electrochemical properties poly(AHNSA)/GCE
Poly(AHNSA) film formation was carried out by the electropoly-
merization of AHNSA monomers using cyclic voltammetry at GCE
in the potential range of
0.8 V and 2.0 V at a scan rate of
0.1 V s
1 for fifteen cycles in 0.1 M HNO3 solution. As shown in
Fig. S1, two oxidation peaks (ipa1 and ipa2) and one reduction peak
(ipc) were observed in the first cycle at peak potentials of 0.054 V,
0.61 V and
0.11 V, respectively. With increase in the number of
voltammetric scans, one oxidation peak (ipa3) was gradually
observed at 0.38 V. All the anodic peak and cathodic peak currents
enhanced gradually with increasing in voltammetric scans. The
increment in the intensity of the peak currents with increasing
scanning cycles confirms the formation and growth of polymer
film onto GCE. Furthermore, the electrochemical behavior of the
modified electrode was studied using cyclic voltammetry between

0.8 V and +0.8 V in 0.5 M H2SO4 solution without the monomer
(inset Fig. S1). Three couples of redox peaks (ipa1-ipc1, ipa2-ipc2
and ipa3-ipc3) were observed which further confirmed that glassy
carbon electrode has already modified. These results are consistent
to the previous related work [340, 341].
3.2. Electrochemical behavior of caffeine and paracetamol
Fig. 1A shows cyclic voltammograms of 1.0 mM caffeine in pH
5.0 acetate buffer solution at a bare glassy carbon electrode and
at poly(4-amino-3-hydroxynaphthalene sulfonic acid) modified
glassy carbon electrode at a scan rate of 0.1 V s
1. At bare glassy
carbon electrode, caffeine showed electrochemically irreversible
158 M. Tefera et al. / Food Chemistry 210 (2016) 156–162

Fig. 1. Cyclic voltammograms obtained (A) in 0.1 M acetate buffer solution (pH 5.0) at poly(AHNSA)/GCE without caffeine (a), in the presence of 1.0 mM caffeine (c) and at
GCE in the presence of 1.0 mM caffeine (b); (B) without paracetamol (a), in the presence of 0.1 mM paracetamol (c) at poly(AHNSA)/GCE and at GCE in the presence of 0.1 mM
paracetamol (b); (C) 1.0 mM caffeine and 0.1 mM paracetamol in 0.1 M acetate buffer solution at GCE (b) and at poly(AHNSA)/GCE (c), and in the absence of caffeine and
paracetamol at poly(AHNSA)/GCE (a) at a scan rate of 0.1 V s
1.

characteristic with an oxidation peak at 1.60 V (curve b). In com-
parison to the bare glassy carbon electrode, a better peak current
response and a negative shift in potential (to 1.45 V) was observed
for caffeine oxidation at poly(4-amino-3-hydroxynaphthalene
sulfonic acid) modified glassy carbon electrode (curve c). The
oxidation of caffeine at the electrode is also found to be
irreversible, as reported in literature (Guo et al., 2011; Habibi
et al., 2012; Torres et al., 2014).
Similarly, paracetamol showed an irreversible wave with an
oxidation peak at 0.563 V at bare glassy carbon electrode (curve
b Fig. 1B). However, a pair of well-defined redox peaks at the
anodic peak potential of 0.477 V and cathodic peak potential of
0.388 V with peak-to-peak separation of 0.089 V and enhancement
of peak current (two fold) was observed at poly(4-amino-3-hydro
xynaphthalene sulfonic acid) modified glassy carbon electrode
(curve c Fig. 1B).
examined in acetate buffer solution of pH 5.0 containing caffeine
and paracetamol using cyclic voltammetry (Fig. 1C). Caffeine
showed an oxidation peak at 1.53 V with a peak current of 35 lA
at the bare glassy carbon electrode (curve b). However, at the pol
y(4-amino-3-hydroxynaphthalene sulfonic acid) modified glassy
carbon electrode (curve c) the peak current for caffeine oxidation
is enhanced (53 lA) at a potential about 0.1 V less positive than
at the bare glassy carbon electrode. The oxidation peak of paraceta-
mol appeared at +0.62 V without a reduction peak in the reverse
scan at bare glassy carbon electrode (curve b). The hydrolysis of
N-acetyl-p-quinoneimine (NAPQI) to p-benzoquinone is responsi-
ble for the disappearance of the cathodic wave in the cyclic voltam-
mograms (Su & Cheng, 2010).
NHCOCH3 NCOCH3 O
-2H , -2e

3.3. Electrochemical behavior of caffeine and paracetamol mixture

OH O O
The electrochemical performance of poly(4-amino-3-hydroxy Paracetamol NAPQI p-benzoquinone
naphthalene sulfonic acid) modified glassy carbon electrode was
 M. Tefera et al. / Food Chemistry 210 (2016) 156–162 159

In contrast, at the poly(4-amino-3-hydroxynaphthalene This is in agreement with previous reports (Amare & Admassie,
sulfonic acid) modified glassy carbon electrode (curve c) a new 2012; Habibi et al., 2012; Svorc, Tomcik, Svitkova, Rievajm, &
peak was appeared in the reverse direction around 0.46 V. In addi- Bustin, 2012).
tion, the oxidation peak potential shifts to 0.53 V and the peak Similarly, the effect of pH of acetate buffer solution on the
current is twice the corresponding peak at the bare GCE. The electrochemical behavior of paracetamol at poly(4-amino-3-hydro
increase in the oxidation peak currents and the decrease in the xynaphthalene sulfonic acid) modified glassy carbon electrode was
oxidation peak potentials of caffeine and paracetamol with studied using cyclic voltammetry. The peak current increased from
improved reversibility for the latter are clear evidences for the pH 3.0–4.5 and then decreased again with higher pH value
increase the surface area of poly(4-amino-3-hydroxynaphthalene (Fig. 2B). Moreover, both the anodic and cathodic peak potentials
sulfonic acid) modified glassy carbon electrode. The carbonyl group were shifted negatively when the solution pH was increased. The
on caffeine and both carbonyl and hydroxyl groups of paracetamol linear dependence of the peak potential on pH is represented by
form hydrogen bonds with both the hydroxyl and amino groups of the equations:
poly(4-amino-3-hydroxynaphthalene sulfonic acid) and thus,
Epa ðVÞ ¼
0:049pH þ 0:673; R2 ¼ 0:9931
these bonds were responsible for an increased in the adsorption
tendency of both analytes on the surface of poly(4-amino-3-hydro
xynaphthalene sulfonic acid) modified glassy carbon electrode. Epc ðVÞ ¼
0:06pH þ 0:679; R2 ¼ 0:9863
The values of DE/DpH
0.049 and
0.060 showed that equal
3.4. Effect of pH number of electrons and protons (2e
/2H+) were involved, which
is in agreement with previous reports (Atta, Galal, & Azab, 2011;
The influence of the pH on the peak currents and potentials of Kumary et al., 2013; Su & Cheng, 2010).
caffeine were examined by cyclic voltammetry in the pH range
3.5–6.0 at 0.1 V s
1. As depicted in Fig. 2A, the current increases
NHCOCH3 NCOCH3
from pH 3.5 to pH 4.5, and then decreases with an increase in pH.
The peak potential was also affected by a change in pH (inset of
Fig. 2A). The oxidation peak potential of caffeine shifted negatively
+ 2H+ + 2e-
with increasing pH values. A linear relationship was observed
between oxidation peak potential and pH with a regression equa-
tion of E(V) =
0.045pH + 1.67, R2 = 0.9994. A slope of 0.045V/pH
OH O
suggests that the overall process involves transfer of equal number
of protons and electrons (4e
/4H+) as given below:

CH3 The increase in peak current of caffeine and paracetamol at

CH3
CH3 lower pH of acetate buffer solution were due to the interaction
N O
N OH N O
N between the positive charge on caffeine (pKa = 10.4) and paraceta-
3H2O
O
mol (pKa = 9.7) and the negative charge of polymer-modified
+ 4H+ + 4e-
N N surface (pKa  4) (Amare & Admassie, 2012; Atta et al., 2011).
N N
CH3 Therefore, pH 4.5 was chosen as the optimum pH value for further
CH3 O OH CH3
CH3 O studies.

(A) (B)
110

100 1.54
11 0.56
90 1.52 10 0.52 Epa(V) = -0.049 pH + 0.673

80 1.50
9 0.48
2
R= 0.9931
I(μA) = -0.045pH+ 1.67
8
70
2
R = 0.9994 0.44
1.48 7
E(V)
E(V)

60
6.0 3.5 0.40

1.46
6
6.0 3.5 5
0.36
Epc(V) = -0.060 pH + 0.679
50
I(μA)

1.44 4 0.32 2
R= 0.9863
I( μA)

40 3 3.5 4.0 4.5 5.0 5.5 6.0

1.42 pH
2
30
1.40 1
3.5 4.0 4.5 5.0 5.5 6.0
20 pH 0
-1
10
-2
0 -3
-10 -4
0.0 0.2 0.4 0.6 0.8 1.0
0.8 1.0 1.2 1.4 1.6
E(V)
E(V)
Fig. 2. Cyclic voltammograms of (A) 1.0 mM caffeine; (B) 0.1 mM paracetamol in various pH value of acetate buffer at poly(AHNSA)/GCE. Inset: plot of peak currents and peak
potentials versus pH.
160 M. Tefera et al. / Food Chemistry 210 (2016) 156–162

3.5. Effect of scan rate Fig. 4 shows the SWV responses of poly(4-amino-3-hydroxynaph
thalene sulfonic acid) modified glassy carbon electrode for caffeine
The effect of scan rate on the peak current for these two species and paracetamol when the concentrations of these species were
was investigated in acetate buffer solution of pH 4.5 containing increased at the same time. The peak currents for both caffeine
1.0 mM caffeine and 0.1 mM paracetamol. As shown in Fig. 3, peak and paracetamol increased linearly with concentrations in the
currents of both caffeine and paracetamol showed a linear relation- range 10–125 lM. The linear regression equations for caffeine
ship with the scan rate. This confirmed that the electrochemical and paracetamol, respectively are:
behavior of caffeine and paracetamol on the surface of poly(4-am
ino-3-hydroxynaphthalene sulfonic acid) modified glassy carbon
Icaf ðlAÞ ¼ 0:218CðlMÞ þ 5:45; R2 ¼ 0:9989 and
electrode is an adsorption-controlled processes. On the other hand,
for both compounds the peak potentials shifted into positive direc- Ipara ðlAÞ ¼ 0:332C ðlMÞ þ 3:26; R2 ¼ 0:9986
tion with increasing the scan rate. This indicated that the oxidation The detection limit was calculated by using the relationship 3 s/
of caffeine is irreversible and the redox reaction of paracetamol is b, where s is the standard deviation of the blank measured under
quasi-reversible. the same conditions as for the standard sample analysis (n = 5)
and b is the slope of the calibration curve. The detection limits of
3.6. Optimization of square wave voltammetry parameters caffeine and paracetamol were estimated to be 0.79 lM and
0.45 lM, respectively.
The dependence of peak current on square wave step potential,
pulse amplitude and frequency for the response of caffeine and 3.8. Repeatability and stability of the modified electrode
paracetamol in acetate buffer of pH 4.5 were studied. The step
potential was measured in the range of 6–16 mV by fixing the Several measurements were taken to determine the repeatabil-
frequency at 20 Hz and amplitude at 30 mV. The peak height ity of the electrochemical responses of poly(4-amino-3-hydroxy
increases as the step potential increases, exhibiting a maximum naphthalene sulfonic acid) modified glassy carbon electrode for
at 12 mV, after which it decreases accompanied by broadening of 1.0 mM caffeine and 0.1 mM paracetamol in acetate buffer solution
peak width. Therefore, 12 mV was chosen as the optimum square (pH 4.5). The results of ten successive measurements showed a
wave step potential. The amplitude was also optimized in the relative standard deviation of 4.45% and 3.62% for caffeine and
range of 40–100 mV by keeping the frequency and step potential paracetamol, respectively, which indicates that the poly(4-amin
at 20 Hz and 12 mV, respectively. The peak current increased up o-3-hydroxynaphthalene sulfonic acid) modified glassy carbon
to 80 mV and then it decreased. Thus, 80 mV was employed in electrode has good repeatability. The long-term stability of the
the subsequent measurements. Finally, by taking the optimum electrode was evaluated by measuring the response for 1.0 mM
values of amplitude and step potential, the effect frequency on caffeine and 0.1 mM paracetamol after keeping the electrode in a
current response was studied in the range of 20–50 Hz. The peak buffer solution for 15 days. The electrode retained about 95% and
current for caffeine and paracetamol increased up to 35 Hz, but 97% of its original activity for caffeine and paracetamol, respec-
after 35 Hz the peak current decreased. As a result, the 35 Hz tively, indicating the electrode was very stable.
was selected as an optimal value for the subsequent
determination. 3.9. Interference study

3.7. Calibration curve of caffeine and paracetamol The selectivity of poly(4-amino-3-hydroxynaphthalene sulfonic
acid) modified glassy carbon electrode for the determination of
The simultaneous determination of caffeine and paracetamol equimolar concentration (40 lM) of caffeine and paracetamol
was carried out using SWV at the optimized conditions at the pol was investigated in the presence of 40 lM and 400 lM of ascorbic
y(4-amino-3-hydroxynaphthalene sulfonic acid) modified glassy acid, citric acid, glucose, uric acid and Pb2+ by SWV at pH 4.5. The
carbon electrode by varying the concentrations of both analytes. corresponding oxidation peak currents obtained in the presence of

220
200 100
90
I(μΑ) = 244v(V/s) +31.9
180 80
R2 = 0.9979
70
160 60
50
140 40
I(μΑ)

30
Ipa(μA)= 45v (V/s) + 2.3
R2 = 0.9977
0.275 V/s
120 20
10
100 0 Ipc(μΑ) = -34.5 v(V/s) -1.99
R2 = 0.9950
I(μΑ)

-10
80 -20
-30
0.075 V/s
60 0.05 0.10 0.15 0.20 0.25 0.30
v(V/s)
40
20
0
-20
-40
0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8
E(V)

Fig. 3. (A) Cyclic voltammograms of 1.0 mM caffeine and 0.1 mM paracetamol in acetate buffer pH 4.5 at the poly(AHNSA)/GCE at various scan rates (0.075, 0.10, 0.125, 0.150,
0.175, 0.20, 0.225, 0.250 and 0.275 V s
1). Inset: plot of peak current versus scan rate.
 M. Tefera et al. / Food Chemistry 210 (2016) 156–162 161

paracetamol
45
I(μΑ) = 0.332C( μΜ ) + 3.26
40 2
50 R = 0.9986
35 caffeine
30
40
25

I( μΑ)
h
20
I( μΑ) = 0.218C( μΜ) + 5.45
30 2
R = 0.9989
15

10
I(μΑ )

20 5

10
a 0 20 40 60 80 100 120 140
C(μΜ)

-10
0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4
E(V)

Fig. 4. Square wave voltammograms for different concentrations (a = 10, b = 20, c = 30, d = 40, e = 60, f = 80, g = 100 and h = 125 lM) of caffeine and paracetamol at poly
(AHNSA)/GCE. Inset: plot of current versus the concentration of caffeine and paracetamol.

interfering substances were compared with those obtained in the Table 1

absence of each interferents. The percent changes in the peak Recovery test for caffeine and paracetamol in Coca-Cola, Pepsi-Cola and tea samples
(n = 3).
current response of caffeine and paracetamol were less than 5%.
Hence, one can concluded that these species did not significantly Sample Analyte Added Found RSD Recovery
interfere in the determination of caffeine and paracetamol and (lM) (lM) (%) (%)

the modified electrode had good selectivity for the determination Coca-Cola Caffeine 0 2.8 – –
of caffeine and paracetamol. 10 11.7 3.8 89.0
20 20.9 7.6 90.5
Paracetamol 0 – – –
3.10. Analytical application 5 4.7 6.5 94.0
10 10.5 7.5 105.0

The application of the proposed method was examined for the Pepsi-Cola Caffeine 0 3.0 – –
10 12.3 6.4 93.0
determination of both caffeine and paracetamol in Coca-Cola,
20 21.7 6.6 93.5
Pepsi-Cola and tea samples. The samples first were diluted with Paracetamol 0 – – –
the supporting electrolyte and then spiked with known concentra- 5 4.7 6.3 94.1
tions of standard caffeine and paracetamol. A peak for caffeine was 10 10.6 8.7 106.0
observed exactly at the same potentials as that observed in the Tea Caffeine 0 1.2 – –
case of standard caffeine, which indicates the presence of caffeine 20 17.4 10.8 81.0
in all the three samples. The values of caffeine in Coca-Cola, Pepsi- 30 30.5 8.8 97.7
Paracetamol 0 – – –
Cola and tea were found to be 112 (±11) mg L
1, 114 (±6) mg L
1 5 4.5 3.7 90.2
and 13 (±1) mg g
1, respectively, which are in good agreement 20 19.58 2.9 97.9
with values reported in literature (Ali et al., 2012; Yang et al.,
2010; Zhang et al., 2011). However, none of the samples analyzed
contained paracetamol.
In addition, standard solutions of caffeine and paracetamol
were added into the samples to test the applicability of the method Table 2
Comparison of the analytical performance of poly(AHNSA)/GCE for caffeine and
in these matrices using recovery method. The results are summa-
paracetamol determination with previously reported electrodes.
rized in Table 1. The recovery was good, showing that the proposed
methods could be efficiently used for the simultaneous determina- Electrode Analyte Linear LOD Ref.
Range
tion of caffeine and paracetamol in cola and tea samples.
(lM) (lM)
The analytical performance of the present modified electrode
Boron-doped diamond Caffeine 0.78–35 0.096 Jeevagan &
was compared with some other similar modified electrodes
electrode Paracetamol 0.54–61 0.23 John, 2012
(Table 2). These data show that the linear range and detection limit Poly(taurine)/ Caffeine 0.05–100 0.5 Xiong et al.,
were comparable with those of other electrochemical sensors. TiO2-graphen/GCE Paracetamol 0.034 2013
Nafion/ruthenium oxide Caffeine 10–250 1.2 Zen & Ting,
pyrochlore/GCE Paracetamol 5–250 2.2 1997
4. Conclusion Graphite-polyurethane Caffeine 4–200 1.6 Saciloto
screen printed Paracetamol 1–40 0.84 et al., 2013
electrode
In this study, the electrochemical behavior of caffeine and
Poly(AHNSA)/GCE Caffeine 10–125 0.79 This work
paracetamol was investigated at poly(4-amino-3-hydroxynaphtha Paracetamol 0.45
lene sulfonic acid) modified glassy carbon electrode using cyclic
162 M. Tefera et al. / Food Chemistry 210 (2016) 156–162
voltammetry and square wave voltammetry. Poly(4-amino-3-hyd
roxynaphthalene sulfonic acid) modified glassy carbon electrode
exhibited an enhancement in the peak current and shift of the peak
position towards negative potential. Besides, low detection limit,
wide linear range, high selectivity and stability, the simultaneous
determination of caffeine and paracetamol was made possible with
this electrode. Moreover, the electrode was also utilized success-
fully for the analysis of caffeine and paracetamol in Coca-Cola,
Pepsi-Cola and tea samples.
Acknowledgment
We thank the International Science Programe (ISP), Uppsala
University, Sweden for the financial support.
Appendix A. Supplementary data
Supplementary data associated with this article can be found, in
the online version, at http://dx.doi.org/10.1016/j.foodchem.2016.
04.106.
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