Journal of Electroanalytical Chemistry 852 (2019) 113517

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Journal of Electroanalytical Chemistry

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

Simultaneous determination of tartrazine, sunset yellow and allura red in

foods using a new cobalt-decorated carbon paste electrode
Johisner Penagos-Llanos a, Olimpo García-Beltr

n b, **,
an a, Jorge A. Caldero
c d, * c
John J. Hurtado-Murillo , Edgar Nagles , John J. Hurtado
a
Facultad de Ciencias Naturales y Matem
aticas, Universidad de Ibagu
e, Carrera 22 Calle 67, Ibague, Colombia
b
on, Innovaci
on y Desarrollo de Materiales – CIDEMAT, Universidad de Antioquia – UdeA, Calle 70 No. 52-21, Medellín, Colombia
Centro de Investigaci

c
Departament of Chemistry, Universidad de los Andes, Carrera 1 No. 18A-12, 111711, Bogot
a, Colombia
d
an Am
ezaga N
375, Lima, Peru
Departamento de Química Analítica, Facultad de Química e Ingeniería Química, Universidad Nacional Mayor de San Marcos, Calle Germ

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

Keywords: A simple, sensitive and economical sensor based on carbon paste decorated with a cobalt complex (CoC/CPE) was
Tartrazine-sunset yellow developed and applied to the simultaneous detection of pairs of synthetic dyes most commonly used in foods,
Tartrazine-allura red which are tartrazine-sunset yellow (TZ-SY) and tartrazine-allurared (TZ-AR). Micro composite surfaces were
Carbon paste
characterized by electrochemical impedance spectroscopy (EIS). Anodic peak currents for TZ-SY and TZ-AR as a
Cobalt complex
function of pH, CoC concentration, time and adsorption potential were observed at 1.0–0.79 V and 1.0–0.81 V,
respectively, by CV. In addition, the increase in the anodic peak currents was greater than 100% for TZ-SY and
TZ-AR using the carbon paste decorated with the Co(II) complex. The detection limits were 0.30  106 -
0.90  106 mol L1 for the TZ-SY pair and 0.8  107-0.37  106 mol L1 for the TZ-AR pair. The new method
was validated with a standard of unflavored gelatin spiked with small amounts of TZ-SY and TZ-AR and showed a
recovery percentage greater than 90%. Finally, the versatility of the sensor was applied in foods such as orange
and strawberry flavored gelatin, powdered, soft drinks and pharmaceutical dosages. The statistical study showed
that this method is equally reproducible, stable and accurate compared to other previously reported methods.

1. Introduction
The most solid evidence about the importance and increased devel-
opment of new methodologies to detect synthetic dyes such as tartrazine,
sunset yellow and allurared in food is clear from the number of reviews
published in recent years focused on describing new reports and ad-
vances in materials applied to this objective. Some of these reviews
describe the use of spectrophotometric techniques where the detection
limits reported between 1990 and 2010 rangedfrom1.1  105-
1.8  104 mol L1 [1]. Moreover, encompassing chromatography tech-
niques and different extraction methods, approximately fifty references
have been reported up to 2010 for detecting different types of samples in
foods [2,3]. Furthermore, Lipskikh et al., reported approximately 100
articles up to 2018 for the electroanalytical detection of synthetic dyes
[4]. Mercury electrodes are the most sensible electrodes used for
detecting TZ, SY and AR by reduction, exhibiting relatively less inter-
ference from organic compounds, with detection limits from
1.7  109-8.5  109 mol L1 [5–7]. Regarding the use of more envi-
ronmentally friendly electrodes, bismuth and antimony film electrodes
have been reported for the detection of TZ, SY and AR, with detection
limits near that of the mercury electrode [8–12]. With the use of more
complex modified electrodes using metal ion nanoparticles and organic
substances such as ZnO/cysteic acid, ZnO/poly(p-aminobenzenesulfonic
acid), TiO/graphene, Cu/organic frameworks, Au/graphene, Tb/poly(3,
4-ethylenedioxythiophene), bimetallic nanoparticle/graphene, Pt/hex-
adecyltrimethyl ammonium bromide and Ni/poly(diallyldimethy-
lammonium chloride) [13–22], it has been possible to develop sensors to
detect and quantify TZ, SY and AR by oxidation between 0.3 and 1.1 V.
These electrodes have shown great selectivity and have not required
sample treatment to eliminate other substances that are easily oxidized at
positive potentials. Additionally, none of these previously mentioned
reports has investigated the simultaneous identification of TZ, SY and AR.
There are very few reports for the simultaneous detection of TZ, SY and
AR. Some reports use spectrophotometric techniques with derivatives

* Corresponding author.
** Corresponding author.
E-mail addresses: andres.calderon@udea.edu.co (J.A. Calder

on), e.nagles.n@unmsm.edu.pe (E. Nagles).

https://doi.org/10.1016/j.jelechem.2019.113517
Received 9 August 2019; Received in revised form 17 September 2019; Accepted 23 September 2019
Available online 6 October 2019
1572-6657/© 2019 Published by Elsevier B.V.
J. Penagos-Llanos et al.
[23,24], with detection limits of 7.5, 4.4, and 4.4  106 mol L1 for TZ,
SY and AR, respectively, and liquid chromatography [25], with detection
limits of 0.56  106, 0.13  106 and 0.8  107 M for TZ, SY and AR.
Moreover, electroanalytical techniques using glassy carbon modified
with polyallylamine resulted in detection limits of 1.8  106, 3.5  106
and 1.4  106 mol L1 for TZ, SY and AR, respectively, while mercury
electrodes gave detection limits of 0.20  107, 0.11  107 mol L1 and
0.13  107 mol L1 [26,27]. All of these reports are motivated by the
impact that these dyes have on health, especially on children, potentially
causing hyperactive behavior and asthma mainly related to allergies
[28]. Although artificial dyes are regulated by organizations such as the
European Food Safety Authority (EFSA), setting consumption at levels of
7.5, 4.0 and 7.0 mg/kg bw/day for TZ, SY and AR, respectively [29–31],
their continuous use in food justifies the development of new method-
ologies that allow for detection and quantification.
There are no reports yet on the simultaneous detection of TZ, SY and
AR using carbon paste composites with metal ions such as cobalt. There
has been only one report to detect AR in this way: by reduction using
Co3O4 [32]. In this context, we previously studied the synthesis and
characterization of cobalt complexes derived from azoles; they have
received considerable attention due to their low cost, easy preparation,
air stability, biological properties and low toxicity [33–37]. Therefore,
we consider that this work presents a potential application for cobalt
complexes in the development of simple, selective and sensitive sensors
for three different dyes.
2. Experimental procedure
2.1. Reagents and instruments
3 1
Standard solutions of TZ-SY and TZ-AM at 1.7  10 mol L and
0.17  103 mol L1 from pure reagent (Sigma-Aldrich) were prepared
with water type A obtained from the Purifier System Wasser lab ASTM
D1193. Only 10.0 mL of each of these standard solutions was used
throughout the study. The system of H3PO4/H2PO  2
4 and H2PO4 /HPO4 at
0.10 mol L1 (Merck) was used as the phosphate buffer solution (PBS). A
DropSens μStat 400 potentiostat (Oviedo, Spain) was used to perform CV
and square wave voltammetry (SWV). EIS was performed using a Versa
STAT 3 potentiostat/galvanostat from Princeton Applied Research (Oak
Ridge, TN, USA).
2.2. Electrodes
Carbon powder (Sigma-Aldrich) supported inside a PVC cylinder
(internal diameter of 2 mm) with a copper wire as the electrical contact
was used as the working electrode, and Ag/AgCl saturated with KCl
(3.0 mol L1) and a platinum wire completed the set of electrodes. pH
measurements were made with an Orion–430 digital pH/mV meter
equipped with a combined pH glass electrode.
2.3. Preparation of [bis(3,5-dimethyl-1-pyrazolyl)methane-NN]cobalt(II)
(CoC)
The cobalt complex [bis(3,5-dimethyl-1-pyrazolyl)methane-NN]
cobalt(II) was prepared by following a method reported in the litera-
ture [37,38]. First, 101.0 mg of bis(3,5-dimethyl-1-pyrazolyl)methane in
3 mL of methanol was combined with 126.0 mg of CoCl2⋅6H2O in 2 mL of
methanol. The reaction mixture was stirred at r. t. for 60 min. The blue
solid was filtered and washed with methanol and diethyl ether.
2.4. Preparation of carbon paste decorated with cobalt complex
microcomposite (CoC/CPE)
The electrode based on carbon paste powder without CoC was pre-
pared initially with 50.0 mg of graphite powder compacted with 20 μL of
paraffin oil into a PVC cavity until it made contact with a copper wire as
2
Journal of Electroanalytical Chemistry 852 (2019) 113517
the electrical contact. The electrode based on carbon paste powder
decorated with CoC was prepared initially with 4.0 mg of CoC and
50.0 mg of graphite powder compacted with 20 μL of paraffin oil into a
PVC cavity until it made contact with a copper wire as the electrical
contact. The surfaces of the electrodes were cleaned using filter paper
until a smooth-looking surface was obtained. It was possible to obtain
three new surfaces with each electrode by removing the excess material
from the surface and repolishing. The optimal amount of CoC was 2.0 mg
with 50.0 mg of carbon powder.
2.5. Sample preparation
Commercial food samples of gelatin and pharmaceutical dosages of
acetaminophen for treating colds in children were purchased from a local
supermarket. Powdered samples, such as fruit-flavored gelatin and
pharmaceutical dosages for treating colds, were first weighed and diluted
10 times with water. Paracetamol for children was not diluted before the
analysis, and an aliquot of 0.250 mL was enough to observe a differen-
tiated background signal. These samples contain a complex matrix of
several substances, such as sugars, 12%protein, fumaric acid, sodium
citrate, ascorbic acid, carboxymethylcellulose, citric acid, aspartame,
phenylalanine, and titanium dioxide. Acetaminophen for children and
pharmaceutical dosages for treating colds contained acetaminophen,
salicylic acid and caffeine. Orange gelatin contained TZ-SY, cherry
gelatin contained AR, and strawberry gelatin contained SY.
2.6. Measurement procedure
The cell was prepared with 9.8 mL of ultrapure water, 0.20 mL of
0.10 mol L1 PBS and 10–100 μL of 0.17  106 mol L1 TZ-SY and TZ-
AR for SWV and 250 μL for CV. All voltammograms were collected by
scanning from 0.6 V to 1.2 V with a scan rate of 0.05 V s1 for CV and a
frequency of 15 Hz and pulse amplitude of 0.05 V at 0.1 V for 30 s for
SWV.
3. Results and discussion
3.1. Electrochemical characterization of the carbon paste electrode
decorated with Co complex, determined by EIS
The electrochemical properties of the carbon paste electrode deco-
rated with CoC were investigated via EIS using
0.10  104 mol L1 K4Fe(CN)6 in 0.10  104 mol L1 KCl as the test
electrolyte. Fig. 1 shows the Nyquist plots of the electrochemical
impedance measurements performed in the K4Fe(CN)6 electrolyte for
both the CPE and CoC/CPE electrodes. The Randles equivalent electrical
circuit used to fit the impedance results are presented in inset of Fig. 1. A
constant phase element (CPE) instead of pure capacitance was used in the
fitting, taking into account the in homogeneity of the electrode surfaces.
Rs is the electrolyte resistance, and CPEdl is the constant phase element
that takes into account the effective capacitance of the electrical double
layer (Cdleff.). Rct is the charge transfer resistance of the redox process,
and ZD is the finite-length diffusion impedance.
The impedance of a simple faradaic reaction without diffusion can be
calculated in terms of a CPE as follows[39]:
Rs
ZðωÞ ¼ Rs þ (1)
1 þ ðjωÞa Q Rct
where α is the exponential term of the CPE, which takes into account the
heterogeneity of the surface. Q is the pseudocapacitance. It represents the
differential capacity of the surface. The CPE parameters α and Q are al-
ways independent of frequency (ω).
The equivalent capacitance of the electrical double layer (Cdleq.) can
be calculated from the CPEdlfollowing to the equation (2) [39], which
takes into account a surface time-constant distribution of the global
J. Penagos-Llanos et al. Journal of Electroanalytical Chemistry 852 (2019) 113517

where RD is the diffusion resistance into the diffusion layer (ohm s1/2),
and B2 denotes the time constant of the diffusion process. B can be related
with the characteristic diffusion length (L), which should be taken as half
of the thickness of the diffusion layer.

L
B¼ (4)
D1=2
Considering the diffusivity (D) of FeCN3/4-
6 to be 7.6  106 cm2 s1
and the values of B from Table 1, the thicknesses of the diffusion layers
(δ) of the CPE and CoC/CPE are 41 μm and 10 μm, respectively. This
means that the modification of the electrode with Co complex diminishes
the diffusion layer and, as consequence, the characteristic time for
diffusion process is also reduced, being the process faster. The diffusion
resistance of the porous electrode can be expressed as follows:
Fig. 1. Nyquist plots of the CPE and CoC/CPE using an electrolyte of
0.10  104 mol L1 Fe(CN)3/4 in 0.10  104 mol L1, circles and lines are Rct δIlim
6 RD ¼ (5)
the experimental and fitted data, respectively. nFACD

where n is the number of electrons exchanged in the redox process, F is

Table 1 the Faraday constant, A is the electroactive area of the electrode, D and C
Values of the parameters used to fit the impedance results using the electrical are the diffusivity and concentration of the electroactive species
circuit shown in the inset of Fig. 1 (FeCN3/4-
6 ) and Ilim. is the limit current of the diffusion process, which at
Parameter Electrode
the open circuit potential can be replaced by the current exchange of the
redox reaction (I0) [40]where I0 ¼ 7.14  106 mA [41]. The electro-
CPE CoC/CPE
active areas of the electrodes (A) can be calculated using equation (5).
Rs (ohm) 382 380 The calculated values of the A for CoC/CPE and CPE were 3.5  108cm2
4.1  105 4.8  103
Rct (ohm)
and 2.4  105cm2, respectively. The specific capacitances are:
CPEdl (S sα) 1.4  106 6.1  106
α 0.84 0.84 31 F cm2 and 1.25 F cm2 for CoC/CPE and CPE electrodes respectively.
Cdleq. (μF) 30.1 1.1 According to the EIS results, the modification of the CPE with the Co
RD (ohm s1/2) 6.7  104 1.5  105 complex improves not only the conductivity but also reduces the diffu-
B (sec1/2) 1.5 0.4 sion time of the CPE. As a consequence of this, a superior electrochemical
Goodness of fit 7.1  104 5.1  105
response and sensitivity of the CoC/CPE can be expected.

impedance of the electrode. 3.2. TZ-SY and TZ-AR activity on CPE and CoC/CPE by CV and SWV

Rs Rct ð1aÞ=a
Cdleq: ¼ Q1=a ð Þ (2)
Rs þ Rct
The effective capacitance (Cdleff.) of the double layer will result after
dividing the Cdleq. by the electroactive surface area (A).
Table 1 presents the values of the parameters of the Randles electrical
circuits (inset Fig. 1) used to fit the experimental electrochemical
impedance results.
The impedance diagrams of the CPE and CoC/CPE exhibited
depressed and open capacitive loops, which can be associated with the
coupling of at less two time constants corresponding to the electric
relaxation of the double layer capacitance (Cdleff.) in parallel with the
charge transfer resistance (Rct) and the open finite-length diffusion
impedance (ZD). The latter takes into account the diffusion of the elec-
troactive species through a finite diffusion layer composed by the carbon
paste material. The electroactive specie (FeCN3/4-
6 ) concentration can be
consider homogeneous in the solution phase due to stirring and the
concentration of the specie just outside the carbon paste coating is fixed
and constant. As can be seen in Table 1, The Rct value of the CoC/CPE is
two orders of magnitude lower than Rct of the unmodified CPE. This
result indicates that the modification of the CPE with Co complex was
successful and that the applied procedure is effective in increasing both
the electrical conductivity and the electrochemical response of the
electrode. According to this result, it can be said that the presence of
cobalt complex in the CPE electrode facilitates the charge transfer in
specific points of the CPE electrode, i. e. the Co complex acts as an
effective depolarizer for the redox reaction.
The open finite-length diffusion impedance (ZD) observed for the CPE
and CoC/CPE can be expressed by equation (3):
1=2
ZDðωÞ ¼ RD ðB2 jωÞ (3)
The activity of the new composites towards the oxidation of the TZ-SY
and TZ-AR pairs was studied by CV and SWV. The results are shown in
Fig. 2, and the values of the anodic peak potential and increases in the
anodic peak currents are summarized in Tables 2 and 3. The anodic peak
currents for TZ, SY and AR were observed between 0.79 and 1.1 V. Only
for SY was a small cathodic peak current observed, with a ΔE of 0.01 V.
The results clearly showed a large increase in the anodic peak currents for
TZ-SY and TZ-AR with the CoC-modified electrodes compared to the CPE
without modification. In addition, the anodic peak potential values did
not change significantly. Therefore, only large effects on the anodic peak
currents were observed. Because the formation of a complex between
cobalt and the dyes is not possible due to the solid state of the compound,
it is possible that the presence of the cobalt complex increases the charge
transport by increasing the electrode surface area and electrostatic
attraction. The anodic peak current observed at 0.27 V by SWV
(Fig. 2B–D) can be attributed the oxidation of Co in the complex. Nor-
mally, the oxidation of Co2þ to Co3þin other complexes is observed be-
tween 0.3 and 0.0 V [42]. Due to the stability of [bis(3,
5-dimethyl-1-pyrazolyl)methane-NN]cobalt(II), higher potentials may
be required for the oxidation of the Co centers. These results are in
agreement with what was observed in the EIS study. The anodic peak
currents increase for each pair TZ-SY and TZ-AR using CV and SWV was
greater than 100%. On the other hand, with SWV, the increased anodic
peak currents were greater at lower concentrations compared to CV.
Therefore, this technique is chosen for the quantification studies.
Regarding the anodic peak current signal for the dyes studied, it can only
be compared with very few reports; for example, when using Co–Ni with
graphene to detect SY, the oxidation signal was observed at 0.85 V [20].
The anodic peak currents for TZ, SY and AR measured with other
modified electrodes using other metal ions, such as Ti and Au, were
observed at more positive potentials [15,18], those with Pd, Au and Zn

3
J. Penagos-Llanos et al. Journal of Electroanalytical Chemistry 852 (2019) 113517

Fig. 2. (A–C) Cyclic voltammograms of TZ-SY and TZ-AR (3.4  105 mol L1) and (B–D) squarewave voltammograms of TZ-SY and TZ-AR (1.7  106 mol L1) with
CPE (black lines) and CoC/CPE (red lines). Conditions: pH 3.2 (PBS) at a scan rate of 0.05 V s1forCV and at 0.1 V for 30s with Eaccof 0.1 V, frequency of 15 Hz, and
pulse amplitude of 0.05 for SWV.

showing an increase in the anodic current with both dyes. It is possible

Table 2
that due to the similarity in their structure and molecular mass, they are
Anodic peak current increases for TZ-SY with CPE and CoC/CPE.
oxidized at the same potential. This similarity makes it impossible to
Electrode CV (μA) % Current SWV (μA) % Current detect both dyes simultaneously.
increased by CV increased by
The behavior of the anodic peak currents and potentials for TZ-SY and
SWV
TZ-AR was studied by varying the scan rate (υ) between 0.01 and
TZ SY TZ SY TZ SY TZ SY
0.114 V s1. For SY and AR, an adsorption-controlled process was
CPE 0.15 0.21 0.44 0.39 observed by a linear relationship between I (μA) vs. υ (V s1) (Fig. 3A),
CoC/CPE 1.56 1.94 90.4 89.2 5.85 8.27 92.3 95.3 and the lnIp vs Lnυ plot (Fig. S1A) showed slope values near 1.0. For TZ, a
diffusion-controlled process was observed by a linear relationship be-
tween I (μA) vs. square root of scan rate υ (V s1) (Fig. 3B), and the lnIp vs
Table 3
Lnυ plot (Fig. S1B) showed slope values near 0.5 [43]. On the other hand,
Anodic peak current increases for TZ-AR with CPE and CoC/CPE. the change inthe anodic peak currents to more positive values with an
increase in the scan rate confirms an irreversible process for the three
Electrode CV (μA) % Current SWV (μA) % Current
increased by CV increased by
dyes. The small current observed in the inverse process for SY and AR
SWV (Fig. 2S) was not observed at lower scan rates, and the large differences
between the anodic and cathodic currents did not confirm a reversible
TZ AR TZ AR TZ AR TZ AR
electrochemical process. Therefore, the systems cannot be considered
CPE 0.25 0.55 0.54 5.62
reversible for SY and AR. A similar situation was reported for SY on glassy
CoC/CPE 1.25 1.69 80.0 67.5 7.5 47.2 92.8 89.0
carbon and was attributed to a chemical process that at a lower scan rate
occurs faster than the electrochemical process, allowing the removal of
presented the same potentials observed in this report [13,14,18] and the oxidized species from the surface near the electrode [43]. A similar
those with Cu and Tb were observed at less positive potentials [16,19]. diffusion-controlled process for TZ is typically observed using metal
The anodic peak currents for TZ, SY and AR observed with these other ion-modified electrodes [14,15], and an adsorption-controlled process
metal ions were between 0.65 and 1.1 V in the same order, SY˂AR˂TZ. In has been reported for SY and AR with metal ion-modified electrodes and
this report, CoC showed a similar capacity as Zn, Au and Pb ions in in the presence of surfactants [18,21,44].
detecting TZ and SY [13,14,18]. On the other hand, the anodic peak To verify whether the number of electrons transferred is similar to
currents for SY and AR were observed at the same potential value, previous reports for these three dyes using modified electrodes, the shift

4
J. Penagos-Llanos et al.
in anodic peak potentials as a function of pH was studied. The results are
shown in Fig. 4. These results clearly show that the anodic current moved
to less positive potentials with an increase in pH between 2.1 and 6.1.
This behavior indicates that protons are involved in the oxidation of the
three dyes. In addition, EpV vs. pH plot (inset Fig. 4A–B) slope values of
0.033, 0.028 and 0.048 V/pH unit were obtained for TZ, SY and AR,
respectively. These values for TZ-SY are almost half of the value of the
Nernst constant 0.059 V/pH unit, implying that for a reversible process,
one electron was transferred. Similar results were reported for TZ-SY
with ZnO on glassy carbon and carbon paste [13,14]. For AR, the slope
value of 0.048 V/pH was close to the theoretical value of 0.059 V/pH,
indicating that two electrons were transferred. This result was also
similar to what was previously reported for AR on carbon paste [32,45].
On the other hand, the maximum anodic peak currents showed major
variation depending on the pH, with a maximum for TZ and SY at pH 3.0
and for AR at pH 4.1. These pH values were chosen as optimal for further
study.
3.3. Calibration curve for quantifying the detection limits of TZ-SY and TZ-
AR and the stability of the CoC/CPE
Square wave voltammograms and the net anodic currents for TZ-SY,
TZ-AR and AR under optimized conditions (pH 3.1 in 0.001 mol L1 PBS,
EADS of 0.0 V, and tADSof60 s) are shown in Fig. 5 with calibration curves
Fig. 3. Effects of scan rate (υ) and square root of scan rate on the anodic peak current of SY, AR (A) and TZ (B). Conditions: CTZ, SY-ARis 34.0  106 mol L1 at pH 3.2
(0.01 mol L1 PBS) with CoC/CPE vs Ag/AgCl.
Fig. 4. Cyclic voltammograms of TZ-SY (A) and TZ-AR (B) (34.0  105 mol L1) at pH values of 2.1, 3.1, 4.1 and 6.1. (inset: the effect of the pH on the anodic peak
potential) using CoC/CPE. Conditions: scan rate 0.05 Vs-1.
5
Journal of Electroanalytical Chemistry 852 (2019) 113517
(inset) for the three dyes between 0.12 and 3.0  106 mol L1. Under
these conditions, four regression equations of Ip(μA) ¼ a þ mCAR were
obtained, where (m) is the slope and (a) is the intercept. The obtained
slopes were as follows: TZ, 0.817 and 0.601CTZ (inset Fig. 5A–B, black
curves); SY, 0.46CSY (inset Fig. 5A, red curve); and AR, 3.85 and 5.79CAR
(inset Fig. 5B–C, red curves). These values showed an order for the
sensitivity of AR > TZ > SY with detection limits of 0.80  107,
0.30  106 and 0.90  106 mol L1, respectively. The detection limit
for AR was affected in the presence of TZ, but for AR without TZ, the
detection limit and linear range was increased (Fig. 5C). It is possible that
a diffusion-controlled process has fewer kinetic limitations than an
adsorption-controlled process. At higher concentrations of
4.0  106 mol L1 of TZ, SY and AR, the anodic peak currents decreased,
possibly due to surface saturation. With respect to the detection limit, this
report is compared with previous reports using electrodes modified with
other complexes. The results summarized in Table 4 clearly show that by
using electrodes prepared with other complexes, lower detection limits
are obtained compared with this report, but there are few reports on the
application to detect these three dyes. The only example is with HMDE
[27], which is an electrode that is not very friendly to the environment.
With a more environmentally friendly electrode, detection limits greater
than 1.0  106 mol L1 was obtained for the three dyes [26]. Therefore,
the electrode in this report has almost the same sensitivity compared to
HMDE, but with the advantage of being more environmentally friendly.
J. Penagos-Llanos et al. Journal of Electroanalytical Chemistry 852 (2019) 113517

The reproducibility of the sensor was evaluated by consecutive
measurements (n ¼ 6) for three days using two electrodes, one for the TZ-
SY pair and another for the TZ-AR pair. The results showed that the
standard deviation (s) with respect to the average value of each dye was
0.025, 0.070 and 0.055 for TZ, SY and AR, respectively, with a coefficient
of variation percentage (% VC) of 5.4, 5.9 and 5.6. These results show an
acceptable variation because reproducing an identical surface using this
type of electrode is not possible. On the other hand, the initial anodic
peak current values were 0.53, 1.17 and 2.46 μA for 2.0  106 mol L1
TZ, SY and AR, respectively. On the third day, these values decreased to
0.47, 1.12 and 2.10 μA for TZ, SY and AR, respectively; the results
showed decreases of 1.8, 1.4 and 2.43% for the dyes. These results show
that the sensor can be used in a three-day interval without considerable
loss of activity. Additionally, the stability and durability of the sensor was
evaluated by performing CV after 25 cycles in standard solutions con-
taining 1.7  106 mol L1 of TZ-SY and TZ-AR. The Fig. 6A show an
anodic peak currents decay of 0.015 and 0.014 μA for the TZ-SY pair and
Fig. 6B show an anodic peak currents decay 0.005 μA for the TZ-AR pair
for each cycle. These results indicate that the performance of the sensor is
fairly solid and it can be used for a long period of time without appre-
ciable reduction of its capacity.

3.4. Validation and interferences of the method

Unflavored gelatin samples spiked with known quantities of TZ-SY

and TZ-AR were prepared to determine the accuracy of the method as
a function of the relative error (RE). One sample (sample 1) was
contaminated with 1.5  106 M TZ-SY, and another sample (sample 2)
was contaminated with 2.5  106 mol L1 TZ-AR. The determination
was carried out using the standard addition method (n ¼ 3), obtaining
(sample 1) 1.43  1060.16 and 1.70  1060.19 mol L1 for TZ and
SY, respectively, with RE values of 4.60 and 23.3%. For TZ-AR, (sample
2) 2.71  1060.20 and 2.82  1060.21 mol L1 was obtained for TZ

Table 4
Electroanalytical methods for TZ, SY and AR.
Dye Modified Method Detection Application Ref.
electrode limit (mol
L1)

TZ 0.2  107
SY HMDE Reduction 0.12  107 Soft drink [27]
powder
–7
AR 0.1310
TZ 1.8  106 Soft drink
powder
SY Polyallylamine/ Reduction 3.5  106 Energy drink [26]
GCE
AR 1.4  106
TZ ZnO-Cys/GCE Oxidation 0.3  107 Soft drink [13]
SY 0.1  107
TZ TiO2-G/CPE Oxidation 0.8  108 Food sample [15]
extracts
SY 0.6  108
TZ Cu-BTC/CPE Oxidation 0.14  109 Soft drink [16]
SY 0.05  109
TZ BiF/SPCE Reduction 3.0  106 Soft drinks, [12]
sweets
6
AR 3.0  10
TZ Zn-p-ABSA/CPE Oxidation 0.8  107 Soft drink [14]
TZ MIP-PmDB/ Oxidation 3.5  109 Soft drinks [46]
PoPD-GCE
SY Aun-G/GCE Oxidation 2.0  109 Soft drink [18]
SY CTAB-G-Pt/GCE Oxidation 4.2  109 Soft drink [21]
AR Co3O4/CPE Oxidation 0.5  107 Soft drink, 32]
energy drink
TZ 0.30  106
SY CoC/CPE Oxidation 0.90  106 Flavored This
gelatin work
powder
AR 0.8  107

CPE: Carbon paste electrode; GCE: Glassycarbonelectrode; ZnO/Cys: Zn oxide/

Fig. 5. Square wave voltammograms and calibration curves (inset) for cysteicacid; G: graphene; BTC: benzenetricarboxylate; p-ABSA: p-amino-
increasing concentrations of TZ-SY (A), TZ-AR (B) and AR (C) with CoC/CPE. benzenesulfonicacid; Aun: goldnanoparticles; CTAB: hexadecyltrimethylammonium-
Conditions: pH 3.1 (PBS) for TZ-SY and TZ-AR and pH 4.1 for AR; EADS 0.10 V; bromide; BiF/SPCE: bismuth film screen-printedcarbonelectrode; MIP-PmDB/
tADS 30 s. PoPD-GCE: Molecularlyimprintedwithdihydroxybenzene and o-phenylenediamine.

6
J. Penagos-Llanos et al. Journal of Electroanalytical Chemistry 852 (2019) 113517

Fig. 6. Cyclic voltammetry curves (25 cycles) on CoC/SPC containing 1.7  106 mol L1 of TZ, SY and AR pH 3.0 (PBS) Scan rate 50 mVs1.

Fig. 7. Square wave voltammograms and calibration curves (insert) for increasing concentrations of TZ-SY (A) and TZ-AR (B) in unflavored gelatin with CoC/CPE.
Conditions: pH 3.1 (PBS) for TZ-SY, pH 4.1 for TZ-AR; EADS 0.10 V; tADS 30 s.

and AR, respectively, with RE values of 8.4 and 12.0%. The highest a 1.5% decrease in the anodic peak currents of TZ, SY and AR.
positive error for SY and AR may be due to the adsorption process
generating a memory effect on the electrode surface. On the other hand,
the unflavored gelatin matrix can be considered a complex matrix, con- 3.5. Analysis of real samples
taining 12% protein, and the RE values for SY and AR were close to 10%.
The only matrix effect of the sample was observed in the shift of the To evaluate the versatility of the proposed method, samples of gelatin
potential to more positive values, and for the TZ-AR pair, a new oxidation powder of different flavors were purchased in local market stores,
signal was observed at 0.72 V at higher concentrations. It is possible that simultaneously containing TZ-SY and TZ-AR and individually containing
this peak is a product of the oxidation of AR that was adsorbed on the TZ, SY and AR. It was not possible to obtain a sample that indicated the
surface of the electrode. Therefore, the stability of the sensor was not simultaneous presence of TZ-AR. The manufacturer of the samples
affected by the matrix effect of the sample. Square wave voltammograms analyzed does not report the quantitative content of the dyes but does
and calibrate curves (inset) for the TZ-SY and TZ-AR solutions (1.5 and
2.5  106 mol L1) are shown in Fig. 7. Table 5
Moreover, substances that can cause considerable interference during Real sample analysis.
the analysis of TZ, SY and AR and that may be present in the same Sample dye Found (mol L1) Slope (m) RDS
samples, such as other dyes including amaranth, Sudan and Ponceau4R,
Strawberry gelatin (powder) SY 9.25 1.855 0.14
were studied in the presence of TZ, SY and AR in concentrations ten times a
Orange gelatin (powder) TZ 9.5  106 0.848 0.10
higher. The results showed that these three dyes do not present any type SY 22.2  106 0.379 0.05
of signal between the range of 0.3 and 1.2 V using CoC/CPE. However, Cherry gelatin (powder) AR 31.0  106 0.546 0.05
these other dyes can slightly affect the signals for TZ, SY and AR, possibly Pax orange flavor (powder) TZ 9.2  106 0.629 0.03
a
Dolex (syrup) AR 2.60  106 3.391 0.20
due to competitive adsorption on the surface of the electrode, producing
a
Squarewave voltammograms and calibration curves (insert) shown in Fig. 8.

7
J. Penagos-Llanos et al.
Fig. 8. Square wave voltammograms and calibration curves (insert) for(A) Orange gelatin (powder) and (B)Dolex (syrup) with CoC/CPE. Conditions: pH 3.1 (PBS)
for TZ-SY, pH 4.1 for AR; EADS 0.10 V; tADS 30 s.
indicate the presence of these dyes: strawberry gelatin with SY, orange
gelatin with TZ-SY, cherry gelatin with AR, and two over-the-counter
medications that are consumed in almost the same amount as foods
and used for colds in children and adults, namely, Pax orange flavor
(powder) with TZ and dolex strawberry flavor (syrup) with AR, both
containing paracetamol (PCM). The results are summarized in Table 5,
and square wave voltammograms with calibration curves for Dolex
(syrup) and orange gelatin are shown in Fig. 8. The results obtained were
almost similar to previous reports in which liquid chromatography for
soft drinks with 58.41  106, 33.18  106 and 35.58  106 mol L1
for AR, SY and TZ respectively. Where the concentration for AR was
higher and for SY and TZ were almost the same [25] and electroanalytical
techniques with 137  106, 18.8  106 and 54.2  106 mol L1 for
AR, SY and TZ respectively [26] for gelatin powder were used. Addi-
tionally, the values obtained are under the European Food Safety Au-
thority (EFSA) norm. With respect to the stability of the sensor in real
samples, it was clearly observed that the oxidation signal for the three
dyes moved to more positive potentials. This finding may suggest that the
matrix of the sample increased the over potential for the oxidation of the
dyes. In addition, the variation in the slope values of the calibration
curves showed in Table 5is evidence of the major effect of the sample
matrices. Despite this effect, the stability of the sensor was not affected,
and it was possible to quantify the dyes in the samples with more than
three points for each sample.
4. Conclusions
This study reported for the first time an electrochemical sensor for the
detection of TZ, SY and AR using a carbon paste electrode and Co com-
plex. The new micro structure was well characterized by EIS and the
electroactive properties were confirmed. A detection limit of less than
1.0  106 mol L1 and only 30 s required per analysis for TZ, SY and AR.
CoC/CPE was highly activity for TZ, SY and AR. The matrix effects were
high but without a considerable loss of sensor activity. Moreover, the
proposed method reported here decreases the time of analysis since it is
not necessary pretreat the samples before each analysis.
Acknowledgments
E. N and O. G. thank the financial support from the Universidad de
Ibagu
e (15-376-INT and 16-416-INT). The authors express their thanks to
Universidad de los Andes for financial support from the Faculty of Sci-
ences (project nos. INV-2018-50-1354 and INV-2017-65-1683) and the
8
Journal of Electroanalytical Chemistry 852 (2019) 113517
Department of Chemistry.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://
doi.org/10.1016/j.jelechem.2019.113517.
References
[1] A. Kauri, U. Gupta, The review on spectrophotometric determination of synthetic
food dyes and lakes, GU J Sci 25 (2012) 579–588.
[2] M. Kucharska, J. Grabka, A review of chromatographic methods for determination
of synthetic food dyes, Talanta 80 (2010) 1045–1051.
[3] K. Yamjala, M. SubramaniaNainar, N. RaoRamisetti, Methods for the analysis of azo
dyes employed in food industry. A review, Food Chem. 192 (2016) 813–824.
[4] O.I. Lipskikh, E.I. Korotkova, P. Ye, J. Khristunova, B.K. Barek, Sensors for
voltammetric determination of food azo dyes, A criticalreview.Electrochim.Acta
260 (2018) 974–985.
[5] A.H. Alghamdi, Determination of Allura Red in some food samples by adsorptive
stripping voltammetry, J. AOAC Int. 88 (2005) 1387–1393.
[6] M. Gomez, V. Arancibia, C. Rojas, C.E. Nagles, Adsorptive stripping voltammetric
determination of Tartrazine and Sunset Yellow in gelatins and softdrink powder in
the presence of cetylpyridinium bromide, Int. J. Electrochem. Sci. 7 (2012)
7493–7505.
[7] S. Combeau, M. Chatelut, O. Vittori, Identification and simultaneous determination
of Azorubin, Allura red and Ponceau 4R by differential pulse polarography:
application to soft drinks, Talanta 56 (2002) 115–122.
[8] A. Kr
olicka, A. Bobrowski, J. Zarębski, I. Tesarowicz, Bismuth film electrodes for
adsorptive stripping voltammetric determination of Sunset Yellow FCF in soft
drinks, Electroanalysis 26 (2014) 756–765.
[9] K. Adsapour-Zeynali, F. Mollarasouli, Bismuth and bismuth-chitosan
modifiedelectrodes for determination of two synthetic food colorants by netanalyte
signal standard addition method, Cent, Eur. J. Chem. 12 (2014) 711–718.
[10] K. Asadpour-Zeynali, M. Aleshi, Electrochemical modification of glassy carbon
electrode by bismuth-chitosan nanosheets for electrocatalytic reduction and
determination of Tartrazine, Portugaliae Electrochim.Acta 32 (2014) 369–379.
[11] B. Claux, B.O. Vittori, Bismuth film electrode as an alternative for
mercuryelectrodes: determination of azo dyes and application for detection in food
stuffs, Electroanalysis 19 (2007) 2243–2246.
[12] J.A. Rodriguez, M.G. Juarez, C.A. Galan-Vidal, J.M. Miranda, E. Barrado,
Determination of Allura Red and Tartrazine in food samples by sequential injection
analysis combined with voltammetric detection at antimony film electrode,
Electroanalysis 27 (2015) 2329–2334.
[13] P.S. Dorraji, F. Jalali, Electrochemical fabrication of a novel ZnO/cysteic acid
nanocomposite modified electrode and its application to simultaneous
determination of sunset yellow and tartrazine, Food Chem. 227 (2017) 73–77.
[14] G. Karim-Nezhad, Z. Khorablou, M. Zamani, P.S. Dorraji, M. Alamgholiloo, M.,
(2017). Voltammetric sensor for tartrazine determination in soft drinks using poly
(p-aminobenzenesulfonicacid)/zinc oxide nanoparticles in carbon paste electrode,
J. Food Drug Anal. 25 (2017) 293–301.
[15] T. Gan, J. Sun, W. Meng, L. Song, Y. Zhang, Electrochemical sensor based on
graphene and mesoporous TiO2 for the simultaneous determination of trace
colourants in food, Food Chem. 141 (2013) 3731–3737.
J. Penagos-Llanos et al.
[16] L. Ji, Q. Cheng, K. Wu, X. Yang, Cu-BTC frameworks-based electrochemical sensing
platform for rapid and simple determination of Sunset yellow and Tartrazine, Sens.
Actuators B 231 (2016) 12–17.
[17] N. Nu~ nez-Dallos, M.A. Macías, O. García-Beltr
an, J.A. Calderon, E. Nagles,
J. Hurtado, Voltammetric determination of amaranth and tartrazine with a new
double-stranded copper (I) helicate-single-walled carbon nanotube modified screen
printed electrode, J. Electroanal. Chem. 822 (2018) 95–104.
[18] J. Wang, B. Yang, H. Wang, P. Yang, Y. Du, Highly sensitive electrochemical
determination of Sunset Yellow based on gold nanoparticles/graphene electrode,
Anal. Chim. Acta 893 (2015) 41–48.
[19] M. Sakthivel, M. Sivakumar, S.M. Chen, K. Pandi, (2018).Electrochemical synthesis
of poly(3,4-ethylenedioxythiophene) onterbium hexacyanoferrate for sensitive
determination of tartrazine, Sens. Actuators B 256 (2018) 195–203.
[20] L. Li, H. Zheng, L. Guo, L. Qu, L. Yu, (2019).Construction of novel electrochemical
sensors based on bimetallic nanoparticle functionalized graphene for determination
of sunset yellow in soft drink, J. Electroanal. Chem. 833 (2019) 393–400.
[21] L. Yu, M. Shi, X. Yue, L. Qu, A novel and sensitive hexadecyltrimethyl ammonium
bromidefunctionalized graphene supported platinum nanoparticles
compositemodified glassy carbon electrode for determination of sunset yellow in
soft drinks, Sens. Actuators B Chem. 209 (2015) 1–8.
[22] L. Yu, M. Shi, X. Yue, X.L. Qu, L., (2016).Detection of allura red based on the
composite of poly (diallyldimethylammonium chloride) functionalized graphene
and nickel nanoparticles modified electrode, Sens. Actuators B Chem. 225 (2016)
398–404.
[23] P.L. Lopez-de-Alba, L. Lopez-Martinez, V. Cerda, L.M. De Leon, Simultaneous
determination of tartrazine, sunset yellow and allura red in commercial soft drinks
by multivariate spectral analysis, Quim. Anal. (Barcelona) 20 (2001) 63–72.
[24] E. Dinç, E. Baydan, M. Kanbur, F. Onur, Spectrophotometric multicomponent
determination of sunset yellow, tartrazine and allura red in soft drink powder by
double divisor-ratio spectra derivative, inverse least-squares and principal
component regression methods, Talanta 58 (2002) 579–594.
[25] Y.S. Al-Degs, Determination of three dyes in commercial soft drinks using HLA/
GOand liquid chromatography, Food Chem. 117 (2009) 485–490.
[26] M.L.S. Silva, M.B.O. Garcia, J. Lima, E. Barrado, Voltammetric determination of
food colorants using a polyallylaminemodified tubular electrode in a
multicommutated flow system, Talanta 72 (2007) 282–288.
[27] P.L. L
opez-de-Alba, L. L

opez-Martinez, L.M. De-Le
on-Rodríguez, Simultaneous
determination of synthetic dyes tartrazine, AlluraRed and sunset yellow by
differential pulse polarography and Partial least squares. A multivariate calibration
method, Electroanalysis 14 (2002) 197–205.
[28] L.J. Stevens, T. Kuczhek, J.R. Burgess, M.A. Stochelski, L.E. Arnold, L. Galland,
Mechanisms of behavioral, atopic, and other reactions to artificial food colors in
children, Nutr. Rev. 71 (2013) 268–281.
[29] European Food Safety Authority (EFSA), Scientific opinion on the re-evaluation
tartrazine (E 102), EFSA J. 7 (2009) 1331.
[30] European Food Safety Authority (EFSA), Reconsideration of the temporary ADI and
refined exposure assessment for Sunset Yellow FCF (E 110), EFSA J. 12 (2014)
3765.
[31] European Food Safety Authority (EFSA), Refined exposure assessment for allura red
AC (E 129), EFSA J. 13 (2015) 4007.
Journal of Electroanalytical Chemistry 852 (2019) 113517
[32] J. Penagos-Llanos, O. García-Beltr
an, J.A. Calder
on, E. Nagles, J. Hurtado, Carbon
paste composite with Co3O4 as a new electrochemical sensor for the detection of
allura red by reduction, Electroanalysis 31 (2019) 695–703.
[33] L. Hern
andez, Y. Quevedo-Acosta, K. V
azquez, A. G
omez-Trevi~ no, J. Zarate-Ramos,
M.A. Macías, J.J. Hurtado, Study of the effect of metal complexes on morphology
and viability of embryonated Toxocaracanis eggs, Vector Borne Zoonotic Dis. 18
(2018) 548–553.
[34] D. Fonseca, C. P
aez, L. Ibarra, P. García-Huertas, M.A. Macías, O. Triana-Ch
avez,
J.J. Hurtado, Metal complex derivatives of bis(pyrazol-1-yl)methane ligands:
synthesis, characterization and anti-Trypanosomacruzi activity, Transit. Met. Chem.
44 (2019) 135–144.
[35] R.A. Murcia, S. Leal, S.M. Roa, E. Nagles, A. Mu~ noz-Castro, J. JHurtado,
Development of antibacterial and antifungal Triazole Chromium(III) and cobalt(II)
complexes: synthesis and biological activity evaluations, Molecules 23 (2018)
2013–2029.


[36] N.J. Bello-Vieda, H.F. Pastrana, M. Garavito, A.G. Avila, A.M. Celis, S. Restrepo,
A. Mu~noz-Castro, J.J. Hurtado, Antibacterial activities of azole complexes combined
with silver nanoparticles, Molecules 23 (2018) 361–378.
[37] K. Castillo, N.J. Bello-Vieda, N.G. Nu~nez-Dallos, F. Homero, Pastrana, A.M. Celis,


S. Restrepo, J.J. Hurtado, A.G. Avila, Metal complex derivatives of azole: a study on
their synthesis, characterization, and antibacterial and antifungal activities,
J. Brazilian Chem. Soc. 27 (2016) 2334–2347.
[38] M.A. Mesubi, B.A. Omotowa, Coordination chemistry of poly(1-pyrazolyl)alkanes
Part V: preparation and characterization of cobalt(II) complexes with bis(1-
pyrazolyl)methane andbis(3-methylpyrazolyl)propane, Transit. Met. Chem. 16
(1991) 348–351.
[39] B. Hirschorn, M.E. Orazem, B. Tribollet, V. Vivier, I. Frateur, M. Musiani,
Determination of effective capacitance and film thickness from constant-phase-
element parameter, Electrochim. Acta 55 (2010) 6218–6227.
[40] A.C. West, Comparison of modeling approaches for a porous salt film,
J. Electrochem. Soc. 140 (1993) 403–408.
[41] X. Wang, M. Wu, W.R. Tang, Y. Zhu, L.W. Wang, J.Q. Wang, P.G. He, Y.Z. Fang,
Simultaneous electrochemical determination of ascorbic acid, dopamine and uric
acid using a palladium nanoparticle/graphene/chitosan modified electrode,
J. Electroanal. Chem. 695 (2013) 10–16.
[42] S. Meghdadi, K. Mereiter, M. Amirnasr, F. Karimi, A. Amiri, synthesis, crystal
structure and electrochemistry of cobalt(III) carboxamide complexes with amine
and azide ancillary ligands, Polyhedron 68 (2014) 60–69.
[43] P. Sierra-Rosales, C. Berríos, S. Miranda-Rojas, J.A. Squella, Experimental and
theoretical insights into the electro oxidation pathway of azo-colorants on glassy
carbon electrode, Electrochim. Acta 290 (2018) 556–567.
[44] E. Nagles, O. García-Beltr
an, Determination of allura red in the presence of
cetylpyridinium bromide by square-wave adsorptive stripping voltammetry on a
glassy carbon electrode, Anal. Sci. 34 (2018) 1171–1175.
[45] S. Chanlon, L. Joly-Pottuz, M. Chatelut, O. Vittori, J.L. Cretier, Determination of
carmoisine, allura red and Ponceau 4R in sweets and soft drinks by differential
pulse polarography, Journal J. Food Compos. Anal. 18 (2005) 503–515.
[46] X. Zhao, Y. Liu, J. Zuo, J. Zhang, L. Zhu, J. Zhang, Rapid and sensitive
determination of tartrazine using a molecularly imprinted copolymer modified
carbon electrode (MIP-PmDB/PoPD-GCE), J. Electroanal. Chem. 785 (2017) 90–95.