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www.wjpls.org │ Vol 7, Issue 1, 2021. │ ISO 9001:2015 Certified Journal │
58
Dakah et al. World Journal of Pharmaceutical and Life Science
STUDY OF CYTOLOGICAL AND GENETIC DAMAGE INDUCED BY A FOOD
COLORING DYE (TARTRAZINE)
*1Dakah Abdulkarim, 2Alsayed Laith and 2Dweik Ali
1Assistant Professor, Department of Biotechnology and Genetic Engineering, Philadelphia University, Jordan.
2Department of Biotechnology and Genetic Engineering, Philadelphia University, Jordan.
Article Received on 15/11/2020 Article Revised on 05/12/2020 Article Accepted on 25/12/2020
1. INTRODUCTION
Some food colors like tartrazine also known as Yellow 5
or E102 is used in many colored foods and drinks
products, aspirin, vitamins and other substances (El
Keredy, 2017), as well as pharmaceuticals and
cosmetics. Mutagenic and toxicity of tartrazine action
were determined by researches, Moreira Soares and his
colleagues evaluated the potential in vitro cytotoxicity,
genotoxicity and effects on DNA repair of human
lymphocytes exposed to the dye (Soares et al., 2015).
Also other researchers studied the mutagenic action of
tartrazine and indigocarmine in a microbial model and in
mice (Karpliuk et al., 1984). Onion (Allium cepa L.) with
number of chromosomes (2n=16) has relatively large
monocentric chromosomes and reasonable as test
organism for the study of environmental mutagenesis
(Moraes and Jordao, 2001; Patra and Sharma, 2002). Al-
Sabti reported that onion root meristem cells are
sensitive to genetic damage by chemical substances (Al
Sabti, 1989). Many researchers used Allium cepa test to
study chromosome damage, Peter and Tomaz reported
the toxicity and genotoxicity effects of methyl
methanesulphonate and they identified 15 categories of
morphological aberrations like chromosome damage,
chromatid damage and centromere damage (Peter and
Tomaz, 2014). Tripathy and Rao evaluated the potential
genotoxicity of orange red dye at various concentrations
on growing root tip cells of onion by analyzing mitotic
cell division and they observed many types of genotoxic
effects like bridge in anaphase and abnormal uncoiling of
chromosomes during metaphase and anaphase; and
transverse orientation of chromosomes in spindle
apparatus (Tripathy and Rao, 2015). The aim of this
study To evaluate cytological and genetic damage that
induced by food coloring dye (tartrazine) on root tip cells
of onion during mitosis.
2. MATERIALS AND METHODS
Dye
Tartrazine (yellow 5) was selected for this study to assay
its genotoxic affect on A. cepa root tip cells and was
purchased from a local market of Amman.
Allium cepa test
Standard protocol (Tripathy and Rao, 2015) was
followed with slight modifications; the bulbs of onion
were purchased from local vegetable market of Amman.
11 onion bulbs presoaked in distilled water until the roots
grew and became long 1 - 2 cm. Various concentrations
of dye (w/v) were prepared in distilled water: 2
mg/100ml, 1,0.5, 0.25, 0.125, 0.0625, 0.03125,
0.015625, 0.0078125, 0.00390625, and control. The
onion bulbs were transferred from distilled water and
presoaked in various concentrations of dye for 48 hour to
Research Article
ISSN 2454-2229
wjpls, 2021, Vol. 7, Issue 1, 58 - 60.
World Journal of Pharmaceutical and Life Sciences
WJPLS
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SJIF Impact Factor: 6.129
*Corresponding Author: Dakah Abdulkarim
Assistant Professor, Department of Biotechnology and Genetic Engineering, Philadelphia University, Jordan.
ABSTRACT
The Allium test is very good indicator for analyzing genetic damage by chemicals, Tartrazine (yellow 5) was
chosen for this study to assay its genotoxic affect on A. cepa root tip cells. Various concentrations of dye (w/v)
were prepared in distilled water and the highest concentration was 2 mg/100ml, The onion roots were soaked in
various concentrations of dye for 48 hour. After that the cut root tips were fixed in 1:3 aceto-alcohols (Carnoy’s
fixative) and stored in 70% alcohol for microscopically studies. Staining was done in 2% aceto-carmine in 45%
glacial acetic acid (v/v). According to the microscopic analysis only concentration 2 mg/100 ml showed abnormal
division and decrease in mitotic index, where mitotic index in dye treated cells was 5,83% and showed significant
decreasing in value compare with control (13,23%). Also several types of abnormal mitotic cells (AMC) were
observed like c-mitosis, multipolar and Binucleus. And according to this study tartrazine are potential genotoxic
agents in the environment.
KEYWORDS: Genetic damage, Tartrazine, c-mitosis, Binucleus, multipolar.
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Dakah et al. World Journal of Pharmaceutical and Life Science
induce mitotic aberrations and to assess mitotic index by
recording the dividing cells. After 48 hr, the cut root tips
were fixed in 1:3 aceto-alcohols (Carnoy’s fixative) (1
acetic acid : 3 ethanol) for 24 hours. Then stored in 70%
alcohol for future use.
Staining and slide preparation
Squash preparation was adopted following the acid
hydrolysis of cellulosic cell wall in 1 N HCl for 1 hour in
room temperature. Staining was done in 2% aceto-
carmine in 45% glacial acetic acid (v/v). Roots tips were
squashed on a slide by light pressure carefully on cover
slips. Then, The microscopic analysis using the 40X
objective was used to calculate mitotic index and
aberrant cells in metaphase, anaphase, and telophase
cells were tested. The cells that showed fragments,
bridges, laggards, c-mitosis and multipolar were
considered aberrant cells.
The mitotic index was calculated as: MI= (Total dividing
cells / Total observed cells)*100
3. RESULTS AND DISCUSSION
According to the microscopic analysis only
concentration 2 mg/100 ml showed abnormal division
and decrease in mitotic index, while lower concentration
did not show any aberrant cells.
Mitotic Index
Table 1 shows the effect of tartrazine on mitotic index. In
untreated meristematic cells, MI was registered to be
13.23%, While in dye treated cells (2 mg/100ml) MI was
5,83%, the results showed significant decreasing in MI
value. Also the maximum number of dividing cells is
(139) in control cells compare with (67) in treated cells.
The observations of the current study are an indication of
the genetic damage by tartrazine, which is evident from
the lowering of the mitotic index and abnormal cells
division. The decreasing of mitotic index could be have
been accomplished by the inhibition of DNA synthesis at
S-phase (Sudhakar et al., 2001); Epel found full
inhibition of mitosis division when ATP level declining
below the 50% of normal level (Epel, 1963); it may be
assumed that the cell division process depended on
energy, and some chemicals like tartazine effect on ATP
synthesis. And according to Jain and Sarbhoy the
chemical substances which impact sugar and ATP
synthesis creating annoxia condition or by other ways
spend much effect on chromosomes movement (Jain and
Sarbhoy, 1988).
Table 1: Effect of Tartrazine on Mitotic index in root tip cells of Allium cepa.
Dye treatments
Total cells scored
Number of dividing cells
Mitotic index (MI)%
Control
1050
139
13,23%
2mg/100 ml
1149
67
5,83%
Genetic damage and aberrant cells
Several types of abnormal mitotic cells (AMC) were
observed like c-mitosis, multipolar and Binucleus (Fig 1
and 2). C-mitosis was more appearance compare with
other aberrant cells. Binucleus cells that were found at
2mg/100ml concentration could be due to the inhibition
of cell wall development at final stage telophase
(Sudhakar et al., 2001). Tartrazine showed effects like
colchicine mode of action, colchicine inhibits the
formation of spindle fibers and temporarily arrests
mitosis division (Blakeslee and Avery 1937). Multipolar
cells resulted from multipolar spindle and unequal
separation of chromosomes, and many studies showed
that pesticides have the same effect (Mohandas and
Grant 1972).
Figure 1: Effect of Tartrazine (2 mg/100 ml) on mitotic cells in root tip cells of Allium cepa. A and B showed
multipolar.
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Dakah et al. World Journal of Pharmaceutical and Life Science
Figure 2: Effect of Tartrazine (2 mg/100 ml) on mitotic cells in root tip cells of Allium cepa. C: showed cmitosis,
D: showed Binucleus.
4. CONCLUSION
From the above results it is clear that tartrazine are
potential genotoxic agents in the environment.
5. FUNDING
This research is supported by Deanship of Development,
Philadelphia University, Jordan.
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