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Toxicological impact of Amaranth, Sunset Yellow and Curcumin as food coloring agents in albino rats

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Background: In this study the possible toxic effect of prolonged administration of three of the widely used food coloring agents are tested. Methods: Adult female albino rats were administered amaranth (4.7 and 47 mg/kg Body weight) , sunset yellow (31.5 and 315 mg/kg bwt.) and curcumin (15.75 and 157.5 mg/kg bwt.) at doses equal to and 10 times the acceptable daily intake (ADI). Following which liver and kidney parameters (glutathione, lipid peroxides, activities of transaminases and Alkaline phosphatase and urea and creatinine concentration) were measured. Furthermore, the effects of these coloring agents on blood picture and on the development of rat embryo were also studied. Results: Our study revealed no effect of amaranth and sunset yellow on liver and kidney glutathione and lipid peroxide levels. Oral administration of curcumin in its low and high doses for 2 months decreased hepatic lipid peroxide concentration. Colorants also did not alter the liver and kidney function when given at the ADI dose but administration of sunset yellow at doses equaling 10 times ADI increased aminotransferases and that of amaranth at 10 times ADI elevated alkaline phosphatase levels. Administration of amaranth at 10 times ADI dose caused skeletal abnormalities in 25% of the examined foeti.
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© J Pak Med Stud. www.jpmsonline.com Volume 1, Issue 2. July-September, 2011. Page | 43
Key Words: coloring agents; liver; kidney; antioxidants; blood picture; teratology
Toxicological impact of Amaranth, Sunset Yellow
and Curcumin as food coloring agents in albino rats
Mohamed M.Hashem1, Attia H.Atta1*, Mahmoud S. Arbid2, Somaia A. Nada2, Samar M. Mouneir1 and GihanFarag Asaad2
Department of Pharmacology, Faculty of Veterinary Medicine, Cairo University, Giza, Egypt.
1* Department of Veterinary Medicine, Faculty of Agriculture and Veterinary Medicine, Qassim University, Buraydah,
KSA.
Department of Pharmacology, National Research Center, Dokki, Giza, Egypt
Abstract
Background: In this study the possible
toxic effect of prolonged administration of
three of the widely used food coloring
agents are tested.
Methods: Adult female albino rats were
administered amaranth (4.7 and 47 mg/kg
Body weight) , sunset yellow (31.5 and 315
mg/kg bwt.) and curcumin (15.75 and 157.5
mg/kg bwt.) at doses equal to and 10 times
the acceptable daily intake (ADI). Following
which liver and kidney parameters
(glutathione, lipid peroxides, activities of
transaminases and Alkaline phosphatase
and urea and creatinine concentration) were
measured. Furthermore, the effects of these
coloring agents on blood picture and on the
development of rat embryo were also
studied.
Results: Our study revealed no effect of
amaranth and sunset yellow on liver and
kidney glutathione and lipid peroxide levels.
Oral administration of curcumin in its low
and high doses for 2 months decreased
hepatic lipid peroxide concentration.
Colorants also did not alter the liver and
kidney function when given at the ADI
dose but administration of sunset yellow
at doses equaling 10 times ADI
increased aminotransferases and that of
amaranth at 10 times ADI elevated
alkaline phosphatase levels.
Administration of amaranth at 10 times
ADI dose caused skeletal abnormalities
in 25% of the examined foeti.
Conclusion: It could be concluded that
high doses of 47 mg/kg bwt. of amaranth
and sunset yellow could impair hepatic
function. Moreover, based on the results
from this study, amaranth should be
avoided during pregnancy.
Conflicting Interest:
None declared
This article has been
peer reviewed.
Correspondence to
Attia H.Atta1
Department of
Veterinary
Medicine, Faculty
of Agriculture and
Veterinary
Medicine, Qassim
University,Burayda
h,KSA
Article submitted on:
15 April, 2011
Accepted on:
15 June, 2011
Funding sources:
Cairo University,
Giza, Egypt and
National Research
Center, Dokki,
Giza, Egypt.
INTRODUCTION
Food additives are substances intentionally added
to food. These may be natural or synthetic (1).
Food additives are commonly used in processed
foods to improve appearance, flavor, taste, color,
texture, nutritive value and conservation. The
principal classes of food additives are coloring
agents, preservatives, flavors, emulsifiers and
stabilizers (2). Coloring agents, also called
colorants, play a significant role in enhancing the
aesthetic appeal of food. Foods that are
aesthetically pleasing are more likely to be
consumed (3,4). The visual aspect may be an
important factor for the selection of products by
final consumers; an important reason why food
dyes stand out as one of the essential additive
class for food industry in their quest for larger
market shares. Even though all color additives
are alike in terms of the Food and Drug
Administration’s (FDA) regulatory definition,
they are regulated in two classes; color
additives requiring certification (synthetic), and
the color additives exempt from certification
(natural). Despite the importance of food
colorants, there is an ever growing concern
© J Pak Med Stud. www.jpmsonline.com Volume 1, Issue 2. July-September, 2011. Page | 44
about the adverse effects of synthetic food colorants on
human health (5). In Egypt, there has been a sharp
increase in the use of synthetic food colorants during the
past few years and this use of synthetic colorants
particularly in food items is not controlled by any
regulatory body (6). There is a dearth of knowledge
regarding the toxic effect of coloring agents or their
effects on fetal development. Since Amaranth had been
banned from the U.S.A since 1976, there is no recent
data from the FDA about toxicity of this coloring agent.
However, Amaranth is still used in many countries and
is approved for use in Egypt
Sunset yellow is currently banned in Norway and
Finland. In 2008, a European Union EU deal specified
that food and drinks containing any of six artificial
colorings that may be linked to hyperactive behavior in
children, including sunset yellow, will have to carry
warnings . Moreover, the European Food Safety
Authority EFSA decided in 2009 to lower the
Acceptable Daily intake for sunset yellow from 2.5
mg/kg to 1.0 mg/kg bodyweight per day. Moreover, a
study found that mixtures of four synthetic colors plus
the preservative sodium benzoate (E211) cause
increased hyperactivity in humans. Sensitivity reactions
may occur when sunset yellow FCF is mixed with other
synthetic colors.(7)
Curcumin, like many antioxidants, can be a "double-
edged sword", where in lab settings, anti-cancer and
antioxidant effects may be seen alongwith pro-oxidant
effects. Carcinogenic effects of curcumin are inferred to
result from interference with the p53tumor suppressor
pathway, an important factor in many human cancers,
including colorectal cancer. Carcinogenic and lethal
dose 50% (LD50) tests in mice and rats, however, have
failed to establish a relationship between tumorogenesis
and administration of curcumin in turmeric oleoresin at
>98% concentrations. Other in vitro and in vivo studies
suggest that curcumin may cause carcinogenic effects
under specific conditions. These conflicting data about
curcumin prompted us to study the toxicological effects
of curcumin, which is used in Egypt in large quantities.
The present investigation was designed (a) to compare
the antioxidant effects of two synthetic (amaranth and
sunset yellow) and one natural (curcumin) color
additives, (b) to study the effect of these agents on
peripheral blood, liver, and kidney function tests and (c)
effect on fetal development in albino rats.(8)
MATERIALS AND METHODS
Animal Seventy adult female Sprague Dawley albino
rats housed in steel mesh cagesfor 2 months and
provided from the breeding unit of the National
Research Center (Giza, Egypt) were used throughout
this study. Animals were fed on commercial standard
pellets enriched with barley and carrots. Experiments
were performed according to the National
Regulations on Animal Welfare and Institutional
Animal Ethical Committee (IAEC).
Coloring agents:
1) Amaranth dye (E123) was obtained from Merck
(Darmstadt,Germany). The compound was supplied
in a 100g bottle in a powdered form.
2) Sunset Yellow FCF (E110) was obtained from El
Gomhouria Co. (Cairo, Egypt). The compound was
supplied in a pure powder.
3) Curcumin: Natural food colorant obtained from
Sigma- Aldrich, USA company.
All drugs were freshly dissolved (amaranth and
sunset yellow) or suspended with Tween 80
(curcumin) in water and given to rats in a dose of
1ml/100 gram body weight
Animal grouping
Rats were randomly divided into seven equal groups
(10/cage).
Group I (Normal control daily water orally at a dose
of 1ml/100g body weightfor 2 months. Group II
(Amaranth ADI): Amaranth 4.7 mg/kg body weight.
Group III (Amaranth 10 times ADI): Amaranth 47
mg/kg body weight.
Group IV (Sunset Yellow ADI): Sunset yellow 31.5
mg/kg bwt.
Group V (Sunset Yellow 10 times ADI): Sunset
yellow 315 mg/kg bwt.
Group VI (Curcumin ADI): Curcumin 15.75 mg/kg
bwt.
Group VII (Curcumin 10 times ADI):
Curcumin157.5 mg/kg bwt.
Drugs were administered daily for 2 months and
doses were calculated for humans and modified for
rats (9). The acceptable daily intake was used
according to FDA and we used 10 times these doses
to see possible toxic effects.
Blood sampling
Blood samples (3ml) were collected every month
from the retro-orbital plexus of each rat into dry
centrifuge tubes and left to clot at room
temperature. Serum samples were obtained after
centrifugation at 1500 rpm for 10 min, and then kept
in clean epindorf tubes at -20°C until analysis.
(AST, ALT, Alkaline Phosphatase, urea and
creatinine) At the end of the experiment another
sample was taken from each animal where the blood
was taken in clean dry vials containing 2 drops of
3.8% sodium citrate to be used for red blood cells
count (10).
Preparation of tissue homogenate
At the end of 2 months a piece of liver and kidney
were taken freshly from each animal on ice and
homogenized in cold saline.The homogenates were
then centrifuged at 4000 rpm for 5 min using a
cooling centrifuge and the supernatant was used for
determination of reduced glutathione (GSH) and
lipid peroxides concentration.
© J Pak Med Stud. www.jpmsonline.com Volume 1, Issue 2. July-September, 2011. Page | 45
cooling centrifuge and the supernatant was used for
determination of reduced glutathione (GSH) and lipid
peroxides concentration.
[1]-Determination of antioxidant activity:
Reduced glutathione GSH content in liver and kidney
homogenates (20%) was determined by a calorimetric
method (11) with some modifications (12), and the lipid
peroxide content in the liver and kidney homogenates
(20%) was determined by a calorimetric method (13).
[2]-Determination of liver and kidney function
Hepatic transaminase activity (ALT and AST) (14),
Alkaline phosphatase activity (ALP) activity (15),
Serum urea nitrogen level (16) and Creatinine
concentration (17) were determined.
[3]-Determination of blood picture
RBC count, haemoglobin (Hb) percentage (18), packed
cell volume (19), mean corpuscular volume (MCV),
mean corpuscular haemoglobin (MCH), and mean
corpuscular haemoglobin concentration (MCHC) were
determined.
[ 4]-Teratogenic effects
To determine any possible teratogenic effects of
amaranth, sunset yellow FCF and curcumin at doses
equal to, and ten times ADI, forty female and twenty
male albino rats were used where every two females
were placed with a male in a separate cage. In the
following mornings, a vaginal smear was taken to verify
the first day of gestation. Pregnancy was confirmed by
the persistence of di-estrus state for 5 days after mating
and palpable fetal masses in the abdomen on the 5th day
of gestation. Animals were allocated into four groups
and given:
Group I (Normal control): daily water orally at a dose
of 1ml/100g bwt. throughout the experimental period.
GroupII (amaranth 10 times ADI): Amaranth at 47
mg/kg bwt.
Group III (sunset yellow 10 times ADI): Sunset yellow
at 315 mg/kg bwt.
Group IV (curcumin10 times ADI): Curcumin at 157.5
mg/kg bwt.
All drugs were given to rats in a dose of 1ml/100 gram
body weight from the 6th day of pregnancy through to
the 15th day of gestation.
On the 20th day of gestation the females were
anaesthetized by alcohol: chloroform: ether (1:2:3)
mixture and caesarean sections were performed to
determine the effect of the drugs on foetal development.
Morphological examination was performed (20), and
the number of implantation and resorption sites was
counted (21). Two thirds of the foeti obtained from each
female were kept in Bouin's solution for one week to
examine visceral abnormalities (20). The last third were
eviscerated and kept in 95% ethanol for 7 days for
dehydration, and subsequently placed in 2% potassium
hydroxide solution for 24-32 hours. The foeti were
immersed in Mallsch's solution with alizarin red for 24
hours, and were then washed again by Mallsch's
solution. Afterwards, the foeti were preserved by serial
passages in 20, 50, 80 and 100% glycerin solution for
detection of skeletal malformation.
© J Pak Med Stud. www.jpmsonline.com Volume 1, Issue 2. July-September, 2011. Page | 46
Groups
Glutathione
Lipid Peroxide
Liver
Kidney
Liver
Kidney
Control
8.6±0.27
5.44±0.16
132 ±9.82
116.45±7.87
Amaranth (ADI)
7.13±0.31
5.51±0.09
141.89±8.98
115.76±3.86
Amaranth (10
xADI)
7.42±0.49
5.84±0.07
115.17±2.99
124.36±3.46
Sunset Yellow ADI
7.48±0.13
5.6±0.07
113.57±2.94
114.18±3.37
Sunset Yellow (10
xADI)
7.63±0.23
5.85±0.18
137±9.81
126.6±4.45
Curcumin ADI
8.5±0.84
5.7±0.17
107±3.66*
118.08±3.52
Curcumin
(10xADI)
9.23±0.31
5.84±0.23
73.5±3.61*
104.59±5.88
Table 1: Effect of administration of amaranth, sunset yellow and curcumin in doses of ADI and 10 times ADI daily for 2 months on liver
and kidney glutathione (umol/gram tissue) and lipid peroxide (nmol/gram tissue) in albino rats (mean ± SE, n=10 ).
Groups
ALT
AST
Alkaline
Phosphatase
Urea
Creatinine
Control
33.24±1.07
5 9 . 9 ± 1 . 2
110.8±2.33
30.39±1.79
0.61±0.01
Amaranth (ADI)
36.51±2.14
6 0 . 9 8 ± 1 . 4 1
116.82±3.03
32.51±2.22
0.62±0.01
Amaranth (10 xADI)
±
6 4 . 4 8 ± 7 . 0 3
126.73±2.74*
33.74±1.23
0.63±0.01
Sunset Yellow ADI
±
5 9 . 5 6 ± 1 . 6 4
117.31±1.58
32.18±2.06
0.63±0.00
Sunset Yellow (10
xADI)
±*
6 5 . 5 2 ± 1 . 6 4 *
117.05±2.33
33.77±1.57
0.62±0.01
Curcumin ADI
±
5 9 . 7 3 ± 0 . 6 9
110.84±2.13
31.66±2.13
0.61±0.01
Curcumin (10 xADI)
±
6 1 . 7 4 ± 2 . 5 2
111.88±2.82
35.04±1.39
0.62±0.01
Table 2: Effect of amaranth, sunset yellow and curcumin in doses of ADI and 10 times ADI daily for 2 months on liver and kidney
function tests in albino rats (mean ± SE, n=10).
Effect on serum Urea and creatinine
Oral administration of amaranth in doses of 4.7 and 47
mg/kg bwt, sunset yellow in doses of 31.5 and 315
mg/kg bwt, and curcumin in doses of 15.75 or 157.5
mg/kg bwt for two months did not affect serum urea or
creatinine concentration as compared to control group.
(Table 2)
Effect on blood picture
Oral administration of amaranth in doses of 4.7 or 47
mg/kg bwt for 2 months did not affect erythrocyte count,
haemoglobin concentration, packed cell volume %,
mean corpuscular volume, mean corpuscular
haemoglobin, mean corpuscular haemoglobin
concentration or total leucocyte count. Oral
administration of sunset yellow at doses of 31.5 or 315
mg/kg bwt for 2months did not affect erythrocyte count,
haemoglobin concentration, mean corpuscular volume
and mean corpuscular haemoglobin as compared to
control group (Table 3), but oral administration of
sunset yellow in the higher dose of 315 mg/kg bwt for 2
months significantly (p≤0.05) decreased PCV% and
MCHC as compared to control group. Similarly, even
though the oral administration of sunset yellow in doses
of 31.5 mg/kg bwt for 2 months did not affect total
leucocytes count, its larger dose significantly decreased
TLC as compared to control group. Oral
© J Pak Med Stud. www.jpmsonline.com Volume 1, Issue 2. July-September, 2011. Page | 47
Groups
Hb content
g/dl
RBCs count
million/mm3
PCV %
MCV (fl=10-
15)
MCH
(pg=1012)
MCHC %
WBCs
count
103/mm3
Control
14.31±0.81
5.68±0.33
30.68±1.5
54.45±2.76
25.53±1.91
46.93±2.54
11.2+0.39
Amaranth ADI
14.24±0.22
5.64±0.27
30.00±0.77
53.71±2.39
25.53±1.17
47.60±1.25
11.83+0.73
Amaranth 10XADI
13.97±0.94
5.04±0.4
28.67±1.98
57.51±3.35
28.01±1.59
48.75±0.7
12.32+0.42
Sunset Yellow ADI
13.58±0.82
5.71±0.28
28.00±1.19
49.16±1.15
23.86±1.33
48.46±1.97
11.7+0.84
Sunset yellow
10XADI
14.14±0.7
5.08±0.24
25.17±1.38*
50.26±4.10
28.21±2.19
37.53±1.05*
9.54+0.54*
Curcumin ADI
12.92±0.69
5.55±0.4
28.00±0.99*
49.34±3.9
24.19±2.65
47.27±1.98
11.22+0.52
Curcumin 10XADI
12.96±0.5
5.31±0.37
28.00±0.78*
54.30±3.54
25.08±1.59
46.63±2.41
9.93+0.58
Table 3: Effect of amaranth, sunset yellow and curcumin in doses of ADI and 10 times ADI daily for 2 months on blood picture of albino rats
(mean ± SE, n=10)
Drug
No of
Pregnant
Dams
Total No. of
foeti
Growth
retardation
Skeletal
abnormalities
Visceral
abnormalities
Total
abnormalities
No. of foeti
%
No. of foeti
%
No. of
foeti
%
No. of foeti
%
Control
1 ml Distilled
Water/rat
6
55
0
0
0
0
0
0
0
0
Amaranth
47 mg/kg bwt
6
36
10
27.8
9
25
3
8.3
22
61.1
Sunset Yellow
315 mg/kg bwt
6
58
6
10.3
0
0
3
5.17
9
15.5
Curcumin
157.5 mg/kg bwt
6
63
2
3.2
0
0
0
0
2
3.1
Table 4: Effect of amaranth, sunset yellow and curcumin in doses of 47, 315 and 157.5 mg/kg bwt respectively (10X ADI) from day 6 to day 15
on foetal abnormalities in albino rats (n=6 ).
© J Pak Med Stud. www.jpmsonline.com Volume 1, Issue 2. July-September, 2011. Page | 48
Figure 2: Foeti of albino rats treated with
Sunset Yellow in a dose of 315mg/kg bwt
from day 6 to day 15 of gestation showing
dilatation of the renal pelvis compared to
control
Figure 1: Foeti of albino rats treated with (C)
curcumin, (B) Sunset Yellow and (A)
amaranth in doses of 157.5, 315 and 47
mg/kg bwt respectively from day 6 to day 15
of gestation showing growth retardation
Figure 3: Foeti of albino rats treated with
amaranth in a dose of 47mg/kg bwt from
day 6 to day 15 of gestation showing
hypoplasia of heart and lung compared to
control
Figure 4: Foeti of albino rats treated with
amaranth in a dose of 47mg/kg bwt
respectively from day 6 to day 15 showing
incomplete ossification of the skull bones,
aplasia of metacarpal and metatarsal bones
and that of caudal vertebrae.
© J Pak Med Stud. www.jpmsonline.com Volume 1, Issue 2. July-September, 2011. Page | 49
Discussion
In this study the possible toxic effects of prolonged
administration of three of the widely used food coloring
agents are tested. Oral administration of amaranth daily
for 2 months in doses of 4.7 and 47 mg/kg bwt did not
affect either hepatic or renal glutathione as compared to
the control group. However, a previous study has reported
an increase in liver glutathione content after prolonged
oral administration of amaranth daily in a dose of 500
mg/kg bwt (23). This difference could be attributed to the
difference in the dosage regimen. Oral administration of
sunset yellow (31.5 and 315 mg/kg bwt) or Curcumin
(15.7 and 157 mg/kg bwt) daily for 2 months did not
affect either hepatic or renal glutathione as compared to
the control group, which is consistent with similar
observation previously reported for curcumin (24).Oral
administration of curcumin (15.7 or 157 mg/kg bwt) daily
for 2 months decreased hepatic lipid peroxide
concentration as compared to the control group., which is
consistent with a previous study where lipid peroxide was
reported to be significantly decreased by administration
of curcumin (25). In the present and previous studies (26)
curcumin was found to have a strong antioxidant effect.
The normal concentrations of ALT and AST after
administration of curcumin could be attributed to the
membrane stabilization effect of curcumin. However,
Oral administration of sunset yellow in a dose 10 times
the ADI for 2 months significantly (p≤0.05) increases
both ALT and AST levels in the serum. This effect may
be due to increased cell permeability of hepatocytes.
Similar observation was previously reported but these
changes did not affect the antioxidant capacity of the
tissue (27).The lack of effect of the tested compounds on
the ALP activity in this study may be due to a membrane
stabilizing effect of these compounds. However, amaranth
in doses of 47mg/kg bwt (10X ADI) for two months
significantly increased serum ALP as compared to control
group. The enzyme ALP is widely distributed in the
body notably in the intestinal mucosa, kidney tubules,
osteoblasts of bone, liver, placenta and lactating
mammary glands. It is mainly localized at the cell
membranes, where it is associated with transport
mechanism (28). Elevation of serum ALP activity is seen
in case of cholestasis or hepatic carcinoma (29). Any of
these or other, yet to to be determined, causes may be
responsible for this increase in ALP. Urea is a waste
product found in blood and formed by the normal break
down of protein in the liver. Urea is normally removed
from the blood by kidneys and then excreted in the urine.
It however, accumulates in the body with renal failure. In
this study, no effect of the tested coloring agents when
given in doses up to 10 times the ADI for 2 months on
serum urea concentration was seen, indicating no effect
on protein catabolism, or renal function. Similar findings
were reported in a previous study, where no differences
were detected between test and control animals in
serum urea nitrogen concentration after administration
of sunset yellow to pigs at levels of 0 (control), 250,
500 and 1000 mg/kg/day for 98 days (31). In another
study however, after administration of curcumin in a
dose of 2.5 and 5% for F344 rats for 13 weeks, the urea
concentration was lowered as compared to the control
group (32). This difference may be due to the different
animal species, difference in doses, or period of
administration.Oral administration of amaranth, sunset
yellow and curcumin in both low and high doses for
two months did not affect serum creatinine indicating
that the three compounds did not affect kidney function
at the used doses. Our results agreed with previous
results which reported no significant change in serum
creatinine concentration after oral administration of
curcumin and sunset yellow in a dose of 2.5 and 5
mg/kg bwt respectively (calculated as acceptable daily
intake for man) to rats for 1 month (26). Oral
administration of amaranth (4.7 and 47 mg/kg) or
curcumin (15.75 and 157.5 mg/kg) for two months did
not significantly alter the blood picture (RBCs, Hb
concentration, PCV%, MCV, MCH, MCHC and TLC)
as compared to control group. This is consistent with
findings from another study, which reported no changes
in blood picture after giving female rats diets
containing 0.03%, 0.3% and 1.5% amaranth for 64
weeks (33). Oral administration of sunset yellow in a
dose of or 315 mg/kg bwt for two months significantly
decreased PCV%, MCHC and total leucocyte count as
compared to control group. The decrease in MCHC
may be attributed to reduction in PCV%, as in a
previous study (34). Oral administration of amaranth at
doses equaling 10 times its acceptable daily intake to 6
female albino rats from day 6 up to day 15 of gestation,
resulted in growth retardation in 27.8%, visceral
abnormalities (Hypoplasia of heart and lung) in 8.3%,
and skeletal abnormalities (incomplete ossification of
the skull bones, aplasia of metacarpal and metatarsal
bones and aplasia of caudal vertebrae) in 25% of the 36
viable foeti. These results are similar to those of
another study which stated that the toxic effect of
amaranth could be attributable to two metabolites,
sodium naphthionate, which causes sternebral
abnormality in fetuses, and the R-amino salt, which
causes skeletal abnormality (35).
Oral administration of sunset yellow at a dose 10
times the acceptable daily intake from day 6 up to day
15 during gestation period did not significantly result
in foetal abnormalities and only 5.17% showed
dilatation of the renal pelvis. This is in agreement
with the results from another study, which stated that
oral administration of sunset yellow to rats in doses of
© J Pak Med Stud. www.jpmsonline.com Volume 1, Issue 2. July-September, 2011. Page | 50
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1, 10, 30 and 100 times the ADI caused no reproductive
abnormalities (36).Oral administration of curcumin at a
dose 10 times the acceptable daily intake from day 6 up
to day 15 during gestation period did not significantly
result in foetal abnormalities except 3.2% of foeti
showed growth retardation. It was reported in a previous
study that curcumin diet at the concentrations of 1500,
3000 and 10,000 ppm for two successive generations
caused no reproductive abnormalities, however a small
reduction in pre-weaning body weight gain of the
second generation (F2) pups at the highest dose level
was observed (37).
Strengths and limitations of the study:
This study added new information on these coloring
agents . We also examined the teratogenic effect for
administering the acceptable daily intake (ADI) which
was unlike previous studies that used very high doses of
these colorants which were far away from commonly
used doses. This study further provides evidence that
perhaps, ADI and 10 times this dose for both Sunset
Yellow and Curcumin can be used safely in our food
but for both Amaranth even the ADI is harmful as it
caused fetal abnormalities. Further studies need to be
performed to confirm our findings and find a better
estimate of an acceptable upper limit of these agents.
Conclusion
The present study showed that synthetic coloring
agents up to 10 times the acceptable daily intake did
not cause any oxidative stress. The ADI of the tested
compounds did not alter either hepatic or renal
function tests. Increasing the dose of synthetic
coloring agents up to 10 times the acceptable daily
intake caused increase in aminotransferases , ALP
and urea concentration in serum. Moreover, amaranth
at 10 times ADI leads to fetal abnormalities.
Acknowledgement
The authors are indebted to Cairo University and to
the NRC for financial and technical support.
© J Pak Med Stud. www.jpmsonline.com Volume 1, Issue 2. July-September, 2011. Page | 51
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... 3 Group C was injected in ovo with 0.2 ml of Cur extract at a dose of 3 mg/kg eggs (Amalraj et al., 2017). 4 Group D was injected in ovo with 0.2 ml of SY at a dose of 1.575 mg/egg (ADI) (Hashem et al., 2011). 5 Group E was injected in ovo with 0.2 ml of Tz at a dose of 0.375 mg/egg (ADI) (Moutinho et al., 2007;JECFA, 2016). ...
... mg/kg b.wt) at the end of experiment there was significant increase in body weight and no mortality between groups and no malformation were observed. Moreover, Hashem et al. (2011) revealed that oral administration of SY and Cur at dose 10 times the ADI from 6 day up to 15 day during gestation showed no fetal abnormalities. However, growth retardation was recorded in the two groups (3.2% and 10.3% respectively). ...
... This is in agreement with Hashem et al. (2019) who revealed that Tz at 0.45 and 4.5 mg/kg induced skeletal malformations in fetuses including missing coccygeal vertebrae, missing strenebrea, missing hind limbs and irregular ribs. A previous study by Hashem et al. (2011) showed similar findings in which oral administration of SY to pregnant females caused growth retardation and skeletal abnormalities in the metacarpal and metatarsal bones and caudal vertebrae. ...
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Abstract Previous studies have suggested that food dyes are responsible for causing number of health problems. The study under consideration aims to show the possible morphological and skeletal malformation induced due to in ovo administration of sunset yellow (SY) and tartrazine (Tz) with or without curcumin (Cur) during organogenesis of developing chick embryo at doses 1.575mg/egg, 0.375mg/egg and 3mg/kg eggs for SY, Tz and Cur comparing with control. The investigation revealed evident reduction in the weight and length of embryos as well as malformations in feather, head, and limbs. Most of the congenital malformations were seen in SY and Tz injected groups such as short beak, excencephaly, kniked tail and pygostyle, curved scapula and retardation in the degree of ossification were the most evident in endoskeleton malformation. In addition, the length of ossified long bones in SY and Tz groups was affected. Co-administration of Cur with SY and Tz ameliorate the reversed effect of SY and Tz on the shape, length, body weight and skeleton of embryos.
... [41] reported that oral administration of synthetic SY to rats after three days at does 500, 1000 and 2000 mg/kg b.wt/day revealed a number of dose dependent degenerative, inflammatory and proliferative lesions, especially in the liver, kidney and spleen. In addition, Ismail&Sakr [23] and Hashem et al. [42] found that the dose of 47 mg/kg b.wt of food coloring amaranth and SY could impair hepatic function and consequently should be avoided during pregnancy. ...
... [18] 4-Group D was injected in ovo with 0.2 ml of SY at a dose of 1.575 mg/egg. [42] 5-Group E was injected in ovo with 0.2 ml of Tz at a dose of 0.375 mg/egg. [45] 6-Group F was injected in ovo with 0.2 ml of 1:1 mixture of Cur extract and SY at the same doses for both. ...
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Coloring agents have been reported to exert deleterious effects on human health and oxidative stress plays a key role in synthetic color induced hepatotoxicity and nephrotoxicity. The present study focused on evaluating histopathological and immune-histochemical effects of two of the most widely used synthetic food coloring agents and the ameliorative role of curcumin (Cur) against this effects. Fertilized eggs were administrated sunset yellow (SY) (1.575mg/egg) and tartrazine (Tz) (0.375mg/egg) with or without Cur (3mg/egg) during organogenesis of developing chick embryo at a dose equivalent to 14 times the acceptable daily intake (ADI) for synthetic colorants SY and Tz. At the histological level, the liver of embryos of SY and Tz groups displayed diffuse vacuolar degeneration in hepatic parenchyma and the portal area showed sever changes. The kidney showed degenerated tubules at cortex with leucocytes inflammatory cells infiltration. In addition, perivascular edema and hyalinized cell lining the proximal tubules were seen. In regarding, the immune-histochemical level expression of Caspase-3 in cytoplasm of hepatocytes and renal convoluted tubules increased in both groups SY and Tz. Co-administration of Cur, as an antioxidant agent, with SY and Tz showed ameliorative effect in all investigated parameters.
... Eight papers on repeated dose toxicity studies were selected from the literature search (Hashem et al., 2011;Kushawaha and Bharti, 2013;Al Dahhan et al., 2014, Khiralla et al., 2015Mahfouz and Moussa, 2015;Elhalem et al., 2016;Ismail, 2016;Khayyat et al., 2018). The FEEDAP Panel evaluated these studies and noted that some of them (Al Dahhan et al., 2014;Mahfouz and Moussa, 2015;Elhalem et al., 2016;Ismail 2016;Khayyat et al., 2018) displayed changes in kidneys, liver and testes, previously observed in other evaluations. ...
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Following a request from the European Commission, the EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) was asked to deliver a scientific opinion on the safety and efficacy of Sunset Yellow FCF for cats, dogs, ornamental fish, grain-eating ornamental birds and small rodents when used as an additive that adds or restores colour in feedingstuffs. Sunset Yellow FCF is considered safe for the target species at the following concentrations in complete feed: 165 mg/kg for cats, 198 mg/kg for dogs, 733 mg/kg for ornamental fish, 24 mg/kg for grain-eating ornamental birds and 750 mg/kg for small rodents. In the absence of adequate information, the FEEDAP Panel cannot conclude neither on the eyes and skin irritation potential of Sunset yellow FCF, nor on its skin sensitisation potential. Exposure by inhalation is considered to be unlikely. The FEEDAP Panel cannot conclude on the efficacy of the additive.
... They are used in order to prevent color loss during technological processing or coloring of food, beverages, supplements and drugs [3]. Sunset yellow (SY), one of the most common dyes in food and pharmaceutical industry, is used with the sole purpose of making the final product more visually appealing and attractive [4]. It is a synthetic dye that belongs to the group of azo colors [5]. ...
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Industrial processed food, pharmaceutical and cosmetical products contain numerous substances the safety of which has been widely concerned. Potassium sorbate (PS) is a common preservative used in a wide-range of products. Sunset yellow (SY) is one of the most commonly used dye in food and pharmaceutical industry. Several studies suggested cytotoxic and genotoxic potential of SY and PS in different cell lines. These effects can lead to organ damage and cancer development. The aim of this study is to investigate cytotoxic and genotoxic potential of two widely used food additives, SY and PS, in Jurkat cell line. Methodology: PrestoBlue assay was used to assess the cytotoxic potential of SY and PS. For observation of DNA damage, cytostatic and cytotoxic effects, the cytokinesis-block micronucleus cytome (CBMN) assay was performed. Results: Decrease of cell viability in Jurkat cell line was observed after 24-hour exposure to both SY and PS. CBMN assay has revealed significant increase of necrotic cells (P<0.05). Genotoxic biomarkers were in physiological range after 24-hour exposure to both analyzed additives. Conclusion: Our findings suggest that SY, as well as PS have cytotoxic potential in Jurkat cell line, as a result of increased number of necrotic cells. Higher cytotoxic effect was caused by SY compared to PS. However, genotoxic potential was not recorded for any of the food additives analyzed.
... Curcumin (E100) has a very low toxicity and no genotoxicity [23], and its acceptable daily intake is 15.75 mg/kgbw [24]. Although it has low bioavailability, this colourant can induce some adverse effects [25], but also seems to have anticancer and antioxidant properties [26]. ...
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Colour is one of the most relevant organoleptic attributes that directly affects consumers’ acceptance and food selection. However, as food colouring pigments are generally unstable and become modified during processing, in order to maintain or restore product colour uniformity, colourants are added to food products around the world. In this context, although they are still widely used, synthetic food colorants, due to their potential hazards, are being replaced by those obtained from natural origins. Indeed, numerous side effects and toxicities, at both the medium and long-terms—namely allergic reactions, and behavioral and neurocognitive effects—have been related to the use of synthetic colourants, whereas their naturally-derived counterparts seem to provide a somewhat high-quality and effective contribution as a health promoter. In order to further understand the implications of the use of synthetic and naturally derived food colourants, this review aims to provide a synoptical approach to the chemical characteristics, properties, uses and side effects on health of those which are currently allowed and applied during food processing.
... The adverse effects of the SY as a synthetic food coloring agent were also previously investigated by different researchers such as (Hashem et al., 2011) showed that a dosage of 47 mg/kg BW of «amaranth and SY» in rats could alter the hepatic function and had better to be avoided throughout the pregnancy. Likewise, (Sayed et al., 2012) proved the mutagenic action of SY dye at dosage of 0.325 mg/kg BW/ day for 1, 2 or 3 weeks and the protective role of Se and A, C and E vitamins in mice with significant chromosomal aberration values in the hepatic tissues. ...
... -Group (C) was injected in ovo with 0.2 mL of Cur extract at a dose of 3 mg/kg eggs [24] . -Group (D) was injected in ovo with 0.2 mL of SY at a dose of 1.575 mg/egg (ADI) [34] . -Group (E) was injected in ovo with 0.2 mL of Tz at a dose of 0.375 mg/egg (ADI) [14] . ...
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