ISSN

Journal of Basic and Environmental Sciences, 7 (2020) 132-139 Online: 2356-6388

Print: 2536-9202

Research paper Open Access

Spectrophotometric determination of Tartrazine in soft drink powder and
pharmaceutical dosage form
Ayman A. Ali1*, Abeer M.E. Hassan2, M.A. Hussien3, Alaa S. Amin1, Ahmed M. Abdelsalam2
1
Chemistry Department, Faculty of Science, Benha University, Benha, Egypt
2
Chemistry Department, Faculty of Pharmacy, October 6th University, October, Egypt
3
Biochemistry Department, Faculty of Pharmacy, October 6th University, October, Egypt
* Corresponding author: ayman.abdelrazik@fsc.bu.edu.eg
Abstract
Simple and rapid spectrophotometric methods were utilized for the determination of tartrazine (E-102) (TZ) in soft
drink powder and pharmaceutical dosage forms. A study of the absorption spectra of tartrazine coloring agent has been
carried out by studying various parameters such as the effect of pH, type, and volume of surfactant and time. The
optimum analytical parameters and their effects are investigated. Beer’s law is obeyed in the range 2.0–20 µg mL-1 at λmax
= 427 nm. For more accurate analysis, Ringbom optimum concentration range is found to be 5-19 µg mL-1. The molar
absorptivity and Sandell sensitivity were determined. The suggested method was successfully applied to the
determination of the examined coloring agent in soft drink powder and pharmaceutical dosage form.
Keywords: Spectrophotometry, Tartrazine, Coloring agents, Drug, Soft drink powder.
Received; 13 Jun 2019, Revised form; 12 Dec. 2019, Accepted; 12 Dec. 2019, Available online 1 April 2020.
1. Introduction
Synthetic dyes are usually added to foodstuffs and soft
drinks not only to improve appearance, color, and texture
but also to maintain the natural color during process or
storage. Synthetic dyes show several advantages compared
with natural dyes such as high stability to light, oxygen
and pH, color uniformity, low microbiological
contamination, and relatively lower production costs.
However, many of them may exhibit adverse health effects
(allergy, respiratory problems, thyroid tumors, Fig (1): The chemical structure of tartrazine
chromosomal damage, urticaria, hyperactivity, abdominal 2. Experimental
pain, etc.) [1-3]. On the other hand, in some cases, the use 2.1. Materials and reagents
of food dyes is also indicative of foodstuff adulteration Tartrazine E 102 (85%) was obtained from kamena for
such as in their addition to Fruit juices. Thus, the use of synthetic colors company (Kafr Tohorms), Giza, Egypt.
synthetic dyes is strictly controlled by laws, regulations Tang Mango (25 g) produced by Mondelez Egypt Food
and acceptable daily intake (ADI) values [4]. These S.A.E., Egypt. Triton X-100 purchased from Alpha
regulations frame the role of the analytical chemist who Chemika Company, India. Sodium Dodecyl Sulphate
has to test for the levels of dyes added to food. Amaranth (SDS), Cetyltrimethylammonium Bromide (CTAB) ad
(E123), sunset yellow FCF (E110) and Tartrazine (E102) Tween 80 purchased from El Naser Pharmaceutical
are among the synthetic dyes mainly used in non-alcoholic Chemicals Company (ADWIC), Egypt. Antinal tablets
beverages [5-8]and the ADI values are between 0-0.5 mg were purchased from Amoun pharmaceutical Co. S. A. E.,
kg−1 for amaranth, 0-2.5 mg kg−1for sunset yellow and 0- Egypt. All chemicals used in our study were of analytical
7.5 mg kg−1 for Tartrazine. Also, Argentine Alimentary grade with the highest purity possible and utilized without
Code establishes a maximal concentration for non- further purification. Freshly bidistilled water was used in
alcoholic drinks of 50 mg L−1 of amaranth and 100 mg L−1 our study and all solutions were day by day prepared.
for sunset yellow FCF and Tartrazine [9]. Figure (1) shows
the chemical structure of Tartrazine. The main target of the 2.2. The preparation of universal buffer solutions:
present work is to describe the development of simple, A series of universal buffer solutions of pH range 2.68–
cheap and rapid spectrophotometric methods for the 12.05 were prepared by using as recommended by Britton
determination of Tartrazine in food and drug samples. [10].
2.3. The preparation of stock solution:
A stock solution of Tartrazine (TZ, 100 µg mL-1) was
prepared by dissolving 10 mg of Tartrazine in 20 mL of
deionized water in 100 mL measuring flask and then

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completed to the mark with deionized water. The obtained between 10 to 100 µg mL-1 of TAR between 10 to 100 µg
solution kept on the magnetic stirrer for 5 minutes to mL-1 were introduced to adjust the volume 20 mL of 0.2 M
complete the solubility of the dye. sodium hydroxide solution and 100 mL acidic mixture (0.4
2.3. The preparation of tartrazine samples from M) were added. The solution was diluted to the mark with
pharmaceutical drugs and soft drink powder: deionized water. The absorption spectra were recorded
Stock solution of Tartrazine TZ (100 µg mL-1) was between 300 and 650 nm. The spectra of all solutions were
prepared by dissolving an exact weight (10 mg) of TZ in measured against a blank of deionized water with 4 ml of
20 mL deionized water in 100 mL measuring flask on a buffer. The measurements were carried out at 25 ± 0.5 0C.
magnetic stirrer for 5 minutes then completed to the mark For the determination of tartrazine dye, the absorption
with deionized water. The soft drinks were used in this spectra of aqueous solutions of Tartrazine (TZ) were
study such as (Tang Mango 25 g (produced by Mondelez recorded in the wavelength range of 390-630 nm.
Egypt Foods S.A.E), Mountain Dew 330 mL kanz 2.5. The procedure of the standard addition method
(produced by PepsiCo, Egypt) and Miranda Green Apple A typical procedure involves the preparation of several
330 mL kanz (produced by PepsiCo, Egypt). The drug was solutions containing the same amount of unknown and
used in this study such as (Antinal capsules produced by mixed with different amounts of the standard solution of
Amoun Pharmaceutical CO.S.A.E and Oxalepetal tablets Tartrazine in a 10 mL measuring flask. The idea of this
produced by Mash Premiere Badr city). procedure is that the total concentration of the sample is
Accurate weight of powder (Antinal capsule, 2.0 g) the combination of the unknown and the standard and that
dissolved in 15 ml of deionized water on a magnetic stirrer the total concentration varies linearly. The concentration of
for 15 min and diluted to the limited mark in 25 mL unknown determined by the relation between the recording
measuring flask. The obtained solution was filtrated, and absorbance and the concentration of standard tartrazine
the filtrate was considered as the starting sample. The dye.
optimum conditions (pH= 6.54, the volume of the buffer of 2.6. Instrumentation
2.0 mL) for tartrazine dye was applied to the obtained All absorption spectra are made using JASCO V-670
filtrate. The absorbance of the obtained solution was (UV-Visible) Spectrophotometer (JAPAN) with scanning
measured at 427 nm. 3.0 g of Oxalepetal powder dissolved speed (10- 4000 nm/min) and spectral bandwidth (0.1 to
in 25 mL of deionized water on a magnetic stirrer for 15 10 nm) for (UV-Vis region) and equipped with 10 mm
min and diluted to the limited mark in 50 mL measuring matched quartz cells. The pH of all solutions was adjusted
flask. The obtained solution was filtrated, and the filtrate to the required value using pH-Meter type ad1030 Adwa
was considered as the starting sample. The optimum (Romania) and Sigma 3-30 KS Cooling Centrifuge speed
conditions for tartrazine dye were applied to the obtained from (1000-2400 rpm Germany).
filtrate. The absorbance of the obtained solution was 3. Results and discussion
measured at 427 nm.
3.1. Study of the optimum conditions for
Accurate weight of powder (0.5 g of Tang Mango)
determination of tartrazine:
dissolved in 25 mL of deionized water on a magnetic
To estimate the optimum conditions for tartrazine with
stirrer for 15 min then makes dilution to the limited mark
other materials and the effect of different experimental
in 50 mL measuring flask. The obtained solution was
variables were investigated and recorded.
filtrated, and the filtrate was considered as the starting
3.2. The effect of pH
sample. The optimum condition (pH= 6.54, the volume of
For determination of optimum pH for tartrazine, the
the buffer of 2.0 mL) for tartrazine was applied to the
absorbance of the prepared samples measured against the
obtained filtrate. The absorbance of the produced solution
blank within the wavelength range of 800-200 nm. The
was measured at 427 nm. The sample solution prepared by
sample prepared by mixing 1 mL of stock solutions, 4 ml
using different volumes of Miranda green apple varied
of the buffer solution, (universal buffer varies from pH 2
from (1.0 to 8.0 mL) from the bottle, 2.0 mL of buffer
to 12) and completed the solution by deionized water to
(pH= 6.54) and completed with deionized water to the
the limited mark in 10 mL measuring flask. The blank also
mark in 10 mL measuring flask and shake well. The
prepared in the same way without the stock sample
absorbance of the obtained solution was measured at 427
solution. Figure 2(a) shows the absorption spectra of the
nm against the blank. The Mountain dew samples solution
effect of the pH 2.0-12 on the absorption of tartrazine.
was prepared in the same way and similar to Miranda
Figure 2(b) shows the value of absorption of tartrazine
green apple. The absorbance of the obtained solution was
with different pH 2.0-12. Extracted data from Figure 2(b)
measured at 427 nm against the blank.
show that maximum absorption by using buffer solution
2.4. General procedure
with pH 6.54 at λmax= 427 nm and the absorption decreases
In a 10 mL calibrated flask, aliquots of the different
with increasing ph.
reagent stock solutions to obtain final concentration

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Fig (2): Absorption spectra of tartrazine at λmax= 427 nm (a) and the absorption value of tartrazine (b) using a universal
buffer with different pH [pH=2-12].

3.3. Effect of volume of the buffer: obtained solution completed with deionized water to the
limited mark in 10 mL measuring flask with shaking well.
In this part, the effect of buffer volume was studied on
The blank prepared in the same way without the stock
the absorption of tartrazine by the addition of a constant
solution of tartrazine. The absorbances have been
volume of tartrazine 1 mL to a different volume of buffer
measured versus a blank solution at λmax= 427 nm.
solution (1, 2, 3, 4, 5 and 6 mL) with pH 6.54. The

Fig (3): Absorption spectra of tartrazine (a) and the absorption value of tartrazine (b) using various volumes of
universal buffer.

Figure 3(a) shows the absorption spectra of tartrazine tartrazine in the presence of surfactants except Tween 80
with various volumes of buffer solution. Figure 3(b) shows which gives absorbance value near absorbance value of
the relation between the absorption of tartrazine with a solution sample without surfactants.
different value of buffer solution. The extracted data from
Figure 3(b) show that 2 mL gives the maximum absorption
of tartrazine at λmax= 427 nm.
3.4. Effect of surfactants:
In this part, the effect of different types of surfactants
(Triton X-100, CTAB, Tween 80, and SDS) was studied
on the absorption of tartrazine samples. The sample
prepared by mixing 1 mL of tartrazine sample, 2 mL
buffer solution (pH= 6.54) and 1 mL of surfactant (0.5 %).
The obtained solutions were completed by the addition of
deionized water to the limited mark in 10 mL measuring
flask. The blank prepared in the same way without the
sample under study. The absorption of obtained solutions
measured against the blank. Figure (4) shows the effect of
surfactants on the absorption of tartrazine. The data
showed no positive response on the absorption of Fig (4): Absorption spectra of tartrazine without surfactant
and by using different surfactants.

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3.5. Effect of volume of surfactant: prepared in the same way without the tartrazine sample.
From the previous study, the presence of Tween 80 The absorbance measured against the blank for all
surfactant increased the absorption of tartrazine samples. Figure 5(a) shows the absorption spectra of the
comparison with the sample without surfactants at tartrazine sample with different volumes of Tween 80
optimum conditions. In this part, the effect of Tween 80 surfactant. The extracted data from Figure 5(b) show that
volume on the absorbance of tartrazine samples was the addition of 2 mL of Tween 80, producing the best
studied. Sample solution prepared by mixing 1 mL of absorbance of tartrazine. But it was a low absorbance
tartrazine sample, the various volume of Tween 80 (from volume comparison by the sample without surfactants. It is
0.25 to 3 mL) and 2 mL of buffer solution (pH= 6.54). The shown that the presence of various surfactants did not
obtained solution was completed by deionized water to the affect the absorbance value of tartrazine samples.
limited mark in a 10 mL measuring flask. The blank also

Fig (5): Absorption spectra of tartrazine without and with Tween 80 surfactant (a) and the effect of Tween 80 volumes
on the absorbance values of tartrazine (b).

Fig (6): The effect of time on the absorption spectra (a) and the absorbance value with time (b) of tartrazine.

3.6. Effect of time: response on the absorption of tartrazine as shown in

In this part, the effect of time was investigated on the Figures 6(a and b).
absorbance of tartrazine samples at optimum conditions. 3.7. Effect of dye concentration:
Samples prepared by the addition of 1 mL of tartrazine In this part, the effect of concentration of tartrazine was
solution, and 2 ml of buffer solution (pH=6.54). The studied by using the various volume of tartrazine standard
obtained solution was diluted by deionized water to the solution at optimum conditions. Samples solutions
limited mark in 10 mL measuring flask with shake well. prepared by adding different volumes of tartrazine
The absorbance of the prepared samples measured against standard solution to 2 mL universal buffer solution (pH=
the blank. It was found that the time has no positive 6.54). The obtained solution was diluted by deionized

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water to the limited mark in 10 mL measuring flask with

shake well. It found that the absorbance increases with the
addition of tartrazine dyes as shown in Figure 7(a).
3. 8. The validity of Beer's law:
From the extracted data from Figure 7(a) the calibration
graph of Beer's law was constructed by the relation
between the concentration of tartrazine standard samples
and the recording absorbance at optimum conditions as
shown in Figure 7(b). Limits of Beer's law, Sandell
sensitivity, molar absorptivity, and other analytical data
were calculated and listed in Table (1).
3. 9. Ringbom optimum range:
Ringbom method is used for more accurate analysis and
is constructed by plotting the percent transmittance (%T)
and the logarithm of the concentration of the sample under
study [11]. Ayres pointed out that the straight line obtained
in the curve of the law of Beer's does not directly show the
range of concentration by which accurate identification of
colored species can be made [12]. From the extracted data
from Figure 7(a) the standard graph of Ringbom were
constructed by the correlation between the concentration
of tartrazine standard samples and the present
transmittance %T at optimum conditions as shown in
Figure 8.
3. 10. Accuracy and precision
The accuracy and precision of the suggested method
were examined by using different concentrations of the
sample understudy in the range of beer's law and Ringbom
methods. The solutions of the prepared samples were
analyzed in six replicates. The collection data obtained
from this research part are summarized in Tables (2). The
extracted data considered favorable for the concentrations Fig (7): The absorption spectra of tartrazine with various
of the sample under investigation. Tables (3 and 4) show concentrations (0.1 – 6 mL) and application of Beer's law
the evaluation of the accuracy and precision of the for tartrazine using optimum conditions.
proposed method for the determination of tartrazine in
pharmaceutical drug and food samples. Table (5)
introduces a comparison between determinations of
tartrazine by Beer's law and standard addition method in
pharmaceutical drug and food samples.

Fig (8): Ringbom plot for tartrazine by using optimum

conditions.

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Table (1): Analytical data, precision, and accuracy of tartrazine:

Parameters Tartrazine
pH 6.54
Wavelength λmax ( nm ) 427
-1
Beer's law limits (µg mL ) 2 - 20
-1)
Ringbom limits (µg mL 5.0 - 19.95
Molar absorptivity (L mol-1cm-1) 5.377 x 105
Sandell sensitivity (µg cm-2) 9.938 x 10-4
Detection limits (µg mL-1 ) 1.910
-1 )
Quantification limits (µg mL 5.79
Regression equation
Slope 0.05169
Intercept -0.05031
Correlation coefficient 0.992
Student t-test 2.0617
Standard deviation ( SD ) 0.02993
Relative standard deviation (RSD %) 4.628
The standard error of the mean (SEM) 0.0122
Mean value (𝒙
̅) 0.6466
Standard error 0.02477

Table (2): Evaluation of the accuracy and precision of Tartrazine material:

sample Taken (µg mL-1) Found (µg mL-1) Recovery (%)

Tartrazine 8 7.968 99.6

9 8.984 99.82
10 9.686 96.864
12 11.77 98.089

Table (3) Evaluation of the accuracy and precision of the proposed method for determination of tartrazine in
pharmaceutical drugs
Parameters Antinal capsule Oxalepetal
Standard deviation (SD) 0.02089 0.03479
Mean value (𝒙
̅) 0.59586 0.410334
Relative standard deviation (RSD %) 3.5 8.478
Standard error of the mean (SEM) 8.528 x 10-3 0.0142
Student t-test 3.045 2.3039
N 6 6

N= number of observations in the sample equal six replicate determinations

The second part contains the spectrophotometric and the concentration of the sample (Beer's law). The
procedures for determination of tartrazine dye in pure results showed that the best pH= 6.54, the volume of
form, in pharmaceutical drugs and food samples. Some buffer about 2 mL, Tween 80 as a surfactant, 2 mL of
experimental variables are studied for the determination of Tween 80, no effect of time on dye. Beer's law is obeyed
the sample understudy in the pure form such as pH, buffer within the concentration range 2-20 µg mL-1 for tartrazine.
volume, type of surfactant, the volume of surfactant, time For more accurate results, Ringbom optimum

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concentration ranges are determined. Molar absorptivity, the proposed spectrophotometric methods can be applied
Sandell sensitivity, detection, and quantitation limits are routine analysis for determination of the studied materials
calculated. To determine the accuracy and precision of the in pure form, pharmaceutical drugs such as (Antinal
suggested methods, solutions containing different capsule, Oxalepetal 600 mg) which found contain (83.58
concentrations of the studied materials are prepared and and 90.26) µg mL-1, respectively, and food samples such
analyzed in six replicated. The recovery, the relative as (Miranda green apple kanz, Mountain Dew and Tang
standard deviation, the relative error is calculated. mango) also contain about (397.84, 310.56 and 2260.78)
µg mL-1, Respectively. Standard addition method applied
The suggested methods can successfully be applied to to pharmaceutical drugs and food samples and the data
determine the pure material (tartrazine) and their match and near the data from the calibration curve of
pharmaceutical drugs and food samples. The results Beer's law which deduce the validity of the method.
obtained compared to statically by the student's t-test. So
Table (4): Evaluation of the accuracy and precision of the proposed method for determination of tartrazine in food
samples
Parameters Tang mango Mountain dew Miranda green apple
Standard deviation (SD) 0.015355 0.0075 0.0210
Mean value (𝒙
̅) 0.4112 0.25717 0.3435
Relative standard deviation 3.734 2.916 6.1127
(RSD %) -3 -3
Standard error of the mean 6.2686 x 10 3.0618 x 10 8.5732 x 10-3
(SEM)
Student t-test 1.4397 2.5233 1.11988
N 6 6 6

N= number of observations in the sample equal six replicate determinations

Table (5): comparison between determinations of tartrazine by proposed method and standard addition method
Sample name Beer's law curve Standard addition method Standard reference (ADI)

Antinal capsule 34.825 µg/g 34.725 µg/g 7.5 (mg. kg bw-1day-1)

[13]
Oxalepetal 15.043 µg/g 15.0766 µg/g
Tang mango 90.431 µg/g 90.4448 µg/g
Miranda green 1.2055 µg/mL 1.204545 µg/mL
apple
Mountain dew 0.9411 µg/mL mL 0940393 µg/ mL
4. Conclusion:
Simple, accurate, readily available and inexpensive Oxalepetal 600 mg) which found contain (34.825 and
spectrophotometer method was utilized for the 15.043) µg g-1, respectively, and food samples such as
determination of Tartrazine (TZ) in soft drink powder (Miranda green apple kanz, Mountain Dew and Tang
(such as tang mango, mountain dew, and miranda green mango) also contain about (90.431 µg/g, 1.2055 µg mL-1
apple) and pharmaceutical dosage form (such as antinal and 0.9411 µg mL-1), Respectively. Standard addition
capsule and oxalepetal). The experimental conditions were method applied to pharmaceutical drugs and food samples
studied for pure Tartrazine dye and applied to the and the data match and near the data from the calibration
pharmaceutical and food samples. So the proposed curve of Beer's law which deduce the validity of the
spectrophotometric methods can be applied routine method. The results demonstrated that the method is
analysis for determination of the studied materials in pure accurate for Tartrazine dye in pure form and
form, pharmaceutical drugs such as (Antinal capsule, pharmaceutical form and soft drink powder.
Acknowledgments
The authors express many thanks to Benha University, Benha, Egypt for support of the current research.

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