Food Chemistry 138 (2013) 12391242

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

Analytical Methods

A nanosilver-based spectrophotometry method for sensitive determination of

tartrazine in food samples
R. Sahraei a, A. Farmany b, S.S. Mortazavi c,
a

Department of Chemistry, Ilam University, Ilam, Iran

Departments of Chemistry, Hamedan Branch, Islamic Azad University, Hamedan, Iran
c
Young Researchers Club, Hamedan Branch, Islamic Azad University, Hamedan, Iran
b

a r t i c l e

i n f o

Article history:
Received 14 August 2012
Received in revised form 24 October 2012
Accepted 6 November 2012
Available online 16 November 2012
Keywords:
Tartrazine
Silver nanoparticles
Spectrophotometry
Food samples

a b s t r a c t
A new method is reported for sensitive determination of tartrazine in the food samples. The method is
based on the catalytic effect of silver nanoparticle (AgNPs) on the oxidation reaction of tartrazine by
potassium iodate in the acetate buffer medium. The reaction is followed spectrophotometrically by measuring the change in absorbance (DA) at 420 nm using a xed time method (70 s). The reaction variables
were optimised in order to achieve the highest sensitivity. The thirty six criterion detection limit was
0.3 ng/mL, and the relative standard deviation for ten replicate measurements of 30 ng/mL of tartrazine
was 0.98% (n = 10). The method was successfully applied to the determination of tartrazine in lemon, and
papaya-avoured gelatin, candy, and in fruit syrup.
2012 Elsevier Ltd. All rights reserved.

1. Introduction
Synthetic food colourants have been used to improve appearance, taste, avour, and colour of foodstuffs in order to make
them more attractive and appetising. Synthetic dyes have been
used instead of natural colours because of their high stability to
oxidation and heat processes, relatively lower costs, and colour
uniformity. Tartrazine is a synthetic organic food dye that can
be found in common food products such as bakery products, dairy
products, candies, and beverages. The presence and content of
this dye must be controlled due to their potential harmfulness
to human beings. For example, recent studies show that tartrazine has signicant adverse effects on neurobehavioral parameters (Tanaka, 2006). Therefore, the analysis of tartrazine in
foods is important. Until now, different methods such as photometry (Sorouraddin, Rostami, & Saadati, 2011) spectrophotometry
(Berzas, Flores, Llerena, & Farinas, 1999; Sayar & zdemir, 1998;
Vidotti, Cancino, Oliveira, & Rollemberg, 2005; Vidotti & Rollemberg,
2006), chromatography (Al-Degs, 2009; Alves, Brum, de Andrade,
& Netto, 2008; Garcia-Falcon & Simal-Gandara, 2005; Husain
et al., 2006; Ma, Luo, Chen, Sub, & Yao, 2006; Minioti, Akellariou,
& Thomaidis, 2007; Poul, Jarry, Elhkim, & Poul, 2009; Zou, Chen, &

Corresponding author. Tel./fax: +98 811 826 4856.

E-mail address: s.s.mortazavi@iauh.ac.ir (S.S. Mortazavi).
0308-8146/$ - see front matter 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.foodchem.2012.11.029

Shao, 2001; Lancaster& Lawrence, 1999) and electroanalytical

methods (Ghoreishi, Behpour, & Golestaneh, 2012; Kapor,
Yamanaka, Carneiro, & Zanoni, 2001; Silva, Garcia, Lima, & Barrado,
2007) have been reported for the determination of tartrazine.
However, some of these methods are not suitable for routine
monitoring as they are time consuming, complicated and have
poor sensitivity and selectivity. To the best of our knowledge,
no kinetic spectrophotometric method has been reported for the
quantication of the tartrazine.
Kinetic spectrophotometric methods are simple and sensitive
analytical methods which have the advantages of high selectivity,
possibility of no interference from the coloured and/or turbidity
background of the samples, possibility of no interference from
the other active compounds present in the commercial product,
if they are resisting the chemical reaction conditions established
for the proposed kinetic method. In addition, kinetic methods
may come with higher sensitivity in comparison with equilibrium
methods. This contribution describes the development of a simple
kinetic spectrophotometric method for the assay of tartrazine in
food samples. The method is based on catalytic effect of AgNPs
on the oxidation of tartrazine by potassium iodate in acetate
acetic acid medium. It was found that in acetateacetic acid media,
potassium iodate could oxidise the tartrazine resulting in decolouration of the solution. AgNPs can catalysis the reaction. The
difference in absorbance of tartrazine at 420 nm between uncatalysed
and catalysed reaction (DA) is directly proportional to the concentration of tartrazine.

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R. Sahraei et al. / Food Chemistry 138 (2013) 12391242

Table 2
Evaluation of accuracy and precision of the proposed method.

2. Materials and methods

2.1. Apparatus

Sample

All absorbance measurements were carried out on a UVVis

photodiode array spectrophotometer Scincos PDA (Seoul, Korea)
using a quartz cell with an optical path of 1 cm. All pH measurements were made with a pH metre (Metrohm 780, AG CH-9101
Herisau, Switzerland) combined with a glass-calomel electrode.
Transmission electron microscopic images and electron diffraction
pattern of the Ag nanoparticles were obtained using an electron
microscope (model JEM-100CX, JEOL, Tokyo, Japan) at an acceleration voltage of 80 kV.
2.2. Reagents
All chemical used were of analytical reagent grade and were
used without further purication. Double-distilled water was used
throughout the study. A stock solution of tartrazine (103 M) was
prepared by dissolving the appropriate amount of tartrazine in
double-distilled water. Potassium iodate solution (0.01 M) was
prepared by dissolving appropriate amount of potassium iodate
in double-distilled water. Acetateacetic acid buffer (pH = 6) was
used.
2.3. Sample preparation
2.3.1. Powdered gelatin samples
Powdered gelatins (lemon and papaya) of the same brand were
bought in a supermarket in Hamedan (Iran). These gelatins contained sugars, fumaric acid, sodium cyclamate, and sodium saccharin. The preparation instructions for the powdered gelatin box
specied 160 g L1. Un-dyed gelatin was used for validation measurements. Solutions with 1.0 g L1 of gelatin were prepared in
water and aliquots of 0.22.0 mL were used for analysis. The results are given in (Tables 1 and 2).
2.3.2. Fruit syrup sample
Fruit syrup (10.0 mL) was mixed with 20.0 mL of double distilled water. After mixing, the residue was ltered off and ltrate
was diluted with double distilled water up to mark in a 100.0 mL
volumetric ask. The tartrazine content in the above solution
was determined by the recommended procedure and the results
are given in Table 1.

Table 1
Determination of tartrazine in food samples (n = 3).
Sample

Tartrazine added (ng/

mL)

Tartrazine found (ng/

mL)

Recovery
(%)

Lemon powdered gelatin

10.0
20.0
30.0

9.7
19.4
29.9

97.0
97.0
99.6

Papaya powdered gelatin

10.0
20.0
30.0

10.3
19.7
30.1

100.3
98.5
100.3

Fruit syrup
10.0
20.0
30.0

9.8
19.6
29.6

98.0
98.0
98.6

9.7
19.5
29.8

97.0
97.5
99.3

Candy
10.0
20.0
30.0

Tartrazine added
(ng/mL)

Tartrazine found
(ng/mL)

RSD
(%)

Intra-day
Lemon powdered
gelatin
Papaya powdered
gelatin

25.00
50.00
25.00
50.00

24.96
49.88
25.03
49.95

0.13
0.08
0.05
0.10

Inter-day
Lemon powdered
gelatin
Papaya powdered
gelatin

25.00
50.00
25.00
50.00

24.98
50.01
24.97
49.98

0.06
0.09
0.14
0.08

2.3.3. Candy sample

The candy sample (5.0 g) was ground with mortar in a pestle
and subsequently dissolved in double distilled water. After mixing,
the residue was ltered, diluted with double distilled water up to
standard volume. The sample solution was then analysed by the
recommended procedure. The results are given in Table 1.
2.4. Procedure
In a 25 mL test tube, 1.0 mL acetateacetic acid buffer (0.2 M)
with pH = 6, 0.2 mL of Ag-nanoparticle (AgNPs) (100 lM) solution
and 2.0 mL of 0.01 M potassium iodate solution were placed. After
diluting the solution with double-distilled water, the solution was
put in a 1.0 cm quartz cell. The initial absorbance (Ai) at 420 nm
was recorded. After addition of different amounts of tartrazine,
the mixture was equilibrated at room temperature for 70 s. Then
the nal absorbance (Af) was recorded at 420 nm. The absorbance
difference was dened as DA420 = Af  Ai.
2.5. Synthesis of silver nanoparticles
The AgNPs were synthesised in a one-step reduction process in
an aqueous solution. In a typical preparation, a 400 lL aliquot of a
0.1 M AgNO3 aqueous solution was added into 100 mL of an aqueous solution containing 0.10 wt.% of the soluble starch and vigorously stirred for 1 h. The pH of resulting solution was adjusted to
8.0 by adding a 0.1 M NaOH solution. Under this experimental condition, the initial reaction mixture was colourless and the growth
of the AgNPs was monitored at different intervals using UVVis
absorption spectroscopy. After about 1 h, the solution turned light
yellow, which indicated the initial formation of the AgNPs. The
mixture was maintained at 50 C for 24 h and the colour of the
reaction solution became yellow.
3. Results and discussion
3.1. Monitoring the reaction
Fig. 1 shows the UVVis absorption spectra obtained at different
time intervals after mixing AgNO3 aqueous solution with soluble
starch aqueous solution at 50 C. Formation of AgNPs in the colloidal solution was monitored from their absorption spectra as the
small noble metal particles reveal absorption band in the UVVis
spectral region due to surface plasmon resonance (SPR) (Huang &
Yang, 2004). The process of reduction of the Ag ions using the
starch was slow, yielding a broad absorption band centred at about
400 nm until 1 h of reaction, which was assigned to the SPR of
AgNPs. The broadband indicates a relatively high polydispersity,
both in size and shape of the Ag particles. The intensity of the
SPR band increased systematically with the increase of reaction

R. Sahraei et al. / Food Chemistry 138 (2013) 12391242

1241

Fig. 1. Temporal evolution of UVvisible absorption spectra after addition of AgNO3

solution into soluble starch solution at 50 C.

time, to reach a maximum after about 24 h. Thereafter, the intensity of the SPR band did not change. The reduction of Ag ions with
starch aqueous solution at 50 C leads to the formation of AgNPs
that are stable in solution for several months. This indicates that
the soluble starch serves as both reducing and protecting agent.
A typical TEM image of the AgNPs is displayed in Fig. 2a. The
AgNPs are observed with a relatively broad particle size distribution (2085 nm range). The average diameter of the as-prepared
AgNPs is 58 nm with a standard deviation of 5.3 nm. To clarify
the exact crystal structure of the AgNPs, electron diffraction (ED)
measurements were carried out. The diffraction rings of the AgNPs
ED pattern (Fig. 2b) correspond well to the crystalline planes of the
cubic structured Ag, suggesting the nanocrystalline nature of these
AgNPs. The rings in electron diffraction pattern can be assigned to
the [1 1 1], [2 0 0], [2 2 0], [3 1 1], and [2 2 2] crystal planes of a facecentred-cubic (fcc) lattice structure of the AgNPs, respectively (Sun
& Xia, 2002).
Nanoparticles possess a very high surface to volume ratio. This
can be underutilised in areas where high surface areas are critical
for success. This could for example be in the catalytic industry and
some nanoparticles actually have proven to be good catalysts
(Prvulescu et al., 2010). Silver nanoparticles have been tested
for their ability to catalyse the oxidation/reduction of dyes such
as methylene blue (Dong, Zhang, & Zhou, 2010), rose Bengal (Jiang,
Liu, & Sun, 2005) and eosin (Jiang et al., 2005). Recently, catalytically oxidation/reduction of days by AgNPs was studied carefully
(Dong et al., 2010; Jana, Sau, & Pal, 1999; Jiang et al., 2005; Pal,
Sau, & Jana, 1997; Pal, Sau, & Jana, 1998). In this work, tartrazine,
is slowly oxidised to a colourless compound by potassium iodate.
However, in the presence of AgNPs the rate of oxidation strongly
increases. This work is based on the catalytic determination of tartrazine in the presence of AgNPs. Since, AgNPs addition has a catalytic effect on the potassium iodatetartrazine reaction system.
In the other word; the oxidation reaction of tartrazine by potassium iodate in acetateacetic acid media is very slow whereas in
the presence of trace amount of AgNPs, it undergoes a rapid reaction rate. The role of AgNPs as a catalyst in the presence of potassium iodate for the oxidation of tartrazine can be described by the
following reaction,

Tartrazinered

H ;IO
3 ;AgNPs

Tartrazineox

where red and ox are the reduced and oxidised form of tartrazine,
respectively. The reaction showed to be strongly dependent on
the concentration of AgNPs.

Fig. 2. (a) Typical TEM image of the starch-stabilized AgNPs and (b) the electron
diffraction (ED) pattern of the AgNPs.

3.2. Effects of variables

The effect of various buffers, with the same concentration, such
as sulfuric acid, acetateacetic acid buffer, borateNaOH and phosphoric acidNaOH was studied. The result shows that acetate
acetic acid gives greater sensitivity. The optimum value of acetate
buffer concentration is obtained. The results show that 1.0 ml of
acetateacetic acid buffer (0.2 M) with pH = 6 was the best in the
reaction whereas; greater amounts of buffer decreased the sensitivity. This effect is due to the fact that in the presence of higher
concentrations of buffer, the tartrazine is protonated, thus reducing the rate of oxidation reaction.
The effect of AgNPs concentration on the catalytic system is explored. The data obtained were used for the plot of DA versus concentration of AgNPs. The results show that 0.2 mL of AgNPs

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R. Sahraei et al. / Food Chemistry 138 (2013) 12391242

(100 lM) was the best. Greater amount of the AgNPs causes a decrease in the reaction rate.
The effect of iodate concentration on the catalytic determination of tartrazine is studied. The results demonstrate that A420 value increased with an increase in potassium iodate concentration
to 2.0 mL of (0.01 M) potassium iodate concentration. Greater
amounts of potassium iodate decrease the sensitivity. This is due
to the fact that at higher concentrations of potassium iodate, the
rate of blank reaction is very fast and the net reaction rate signal
was diminished.
3.3. Calibration curve & limit of detection (LOD) and interference study
The calibration data for tartrazine were prepared by the procedure described above under the optimum experimental conditions.
There was a linear relationship between A420 and tartrazine concentration in the range of 0.7360 ng/mL with a correlation coefcient of 0.996. The regression equation was A420 = 0.016X (ng/
mL) + 0.010, where X is the tartrazine concentration. The LOD
was calculated according to the recommended formula by International Union of Pure and Applied Chemistry (IUPAC) as LOD = 3SD/
K were SD is the standard deviation of the blank measurements
and K is the slop of the calibration curve. For the method reported
here the LOD was 0.3 ng/mL on the basis of 10 blank
measurements.
In order to investigate the analytical applicability and selectivity, the xed time kinetic method (70 s) was used. A foreign ion
was considered to interfere signicantly when it gave a tartrazine
measurement error of more than 3r. The inuence of several cations and anions were tested using the standard solution of tartrazine (30 ng/mL). The results show that most common ions do not
interfere with the catalytic determination except for Cr3+, Hg2+
and Fe2+ which showed serious positive interferences. However,
the interfering effect of Cr3+and Hg2+ could be eliminated by masking with ethylenediaminetetraacetic acid (EDTA).
3.4. Application
To investigate the applicability of the proposed method, recovery experiment was performed using the standard addition method. For this purpose, a known amount of tartrazine was spiked to
its formulated preparations and the total amount of the dye was
estimated. The results are summarised in Table 1. As can be seen
from the table that the mean recovery ranged from 97.0% to
100.3%.
The accuracy and precision of the method was evaluated by performing three successive measurements within one day at two different concentration levels (25 and 50 ng/mL). The inter day
precision was measured by assaying the bulk sample on ve consecutive days. The results are summarised in Table 2. The results
showed that the RSD (<0.14%) found in intra and inter day assays
can be considered to be very satisfactory.
4. Conclusion
In this contribution, a new sensitive and selective method was
developed for the sensitive quantication of tartrazine based on
the catalytic effect of AgNPs on the oxidation of tartrazine in acetateacetic acid buffer media. The proposed method was applied
to the determination of tartrazine in proprietary foods purchased
from local stores. The results, shown in Tables 1 and 2, suggest that
the method is suitable for the determination of tartrazine. Further-

more, very accurate results were obtained for spiked values of the
tartrazine into the food samples. In addition to accuracy, the method is simple and economical for the determination of tartrazine in
food samples.
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