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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Justification for type of information:
data is from publication.

Data source

Reference
Reference Type:
publication
Title:
Genotoxicity Testing of the Food Colours Amaranth and Tartrazine
Author:
Aparajita Das and Anita Mukherjee
Year:
2004
Bibliographic source:
Int J Hum Genet, 4(4): 277-280 (2004)

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Principles of method if other than guideline:
Ames mutagenicity assay was performed to evaluate the mutagenic nature of the test compound amaranth.
GLP compliance:
not specified
Type of assay:
bacterial gene mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
Name of test material (as cited in study report): Amaranth [Food Red 2]
Molecular formula (if other than submission substance): C20-H11-N2-O10-S3.3Na
C20-H14-N2-O10-S3.3Na
Molecular weight (if other than submission substance): 604.4789 g/mole
Substance type: Organic
Physical state: Solid
Specific details on test material used for the study:
Details on test material
Name of test material (as cited in study report): Amaranth [Food Red 2]
Molecular formula (if other than submission substance): C20H11N2O10S3.3Na
C20H14N2O10S3.3Na
Molecular weight (if other than submission substance): 604.4789 g/mole
Substance type: Organic

Method

Target gene:
Histidine
Species / strain
Species / strain / cell type:
S. typhimurium, other: Salmonella typhimurium TA97a, TA98 and TA100
Additional strain / cell type characteristics:
not specified
Metabolic activation:
without
Metabolic activation system:
no metabolic activation system used
Test concentrations with justification for top dose:
10,100,250,500 and 1000 μg /plate
Vehicle / solvent:
Vehicle
Vehicle(s)/solvent(s) used: Sterile double distilled water
Justification for choice of solvent/vehicle: No data available
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Remarks:
for TA97a and TA98, 20 μg/plate nitro phenylene diamine and for TA100, 1.5 μg/plate sodium azide
Positive control substance:
other: for TA97a and TA98, 20 μg/plate nitro phenylene diamine and for TA100, 1.5 μg/plate sodium azide
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
Preincubation period: No data available
Exposure duration: 48⁰C
Expression time (cells in growth medium): 48⁰C
Selection time (if incubation with a selection agent): No data available
Fixation time (start of exposure up to fixation or harvest of cells): No data available

NUMBER OF REPLICATIONS: triplicate
Evaluation criteria:
Increase in the number of mutagenic revertants
Statistics:
ANOVA test was performed at 0.05 level

Results and discussion

Test results
Species / strain:
S. typhimurium, other: Salmonella typhimurium TA97a, TA98 and TA100
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not valid
Positive controls validity:
valid
Remarks on result:
other: No mutagenic effct were observed

Any other information on results incl. tables

Table 1: Mutagenicity of food colours in tester strains of Salmonella typhimurium

 

Dose

          μg/plate

Mean of the No. Revertant Colonies ± S.D.

TA97a

TA98

TA100

10

120.33 ±

15.50

222.0 ± 49.79

103.7

±

9.07

100

115.67 ±

 17.78

205.7 ±

 4.04

92.7

±

16.17

250

92.33

±

 12.50

170.7

± 68.72

92.0

±

 27.73

500

120.67 ±

22.94

178.3 ± 30.29

83.7

±

 7.095

1000

122.33± 24.82

195.7± 36.23

109.0

±

10.19

10

134.00

±

42.51

49.3

±

13.7

98.0

 ±

7.55

Solvent control

122.30 ±

9.61

21.3

±

9.02

121.0

 ±

 3.61

NPD

827.70 ± 106.53

522.0 ± 50.48

-

SA

-

-

1624.67 ±

 89.76

 

Where:

NPD= Nitrophynylenediamine

SA= Sodium azide

S.D.= Standard Deviation

Anova Value of TA97a (Amaranth —2.11, ns)

TA 98 (Amaranth –10.19*)

TA100 (Amaranth —2.196, ns)

P< 0.01 (5.06)

Applicant's summary and conclusion

Conclusions:
The test compound amaranth [Food Red 2] was not mutagenic in the study conducted using Salmonella typhimurium TA97a, TA98 and TA100 without metabolic activation system.
Executive summary:

Ames mutagenicity assay was performed to evaluate the mutagenic nature of the test compound Amaranth in plate incorporation assay.

 

The test material was tested at a concentration of 10,100,250,500 and 1000 μg /plate. The plates were inverted within an hour and placed in a dark vented incubator at 37⁰C for 48 hours. Positive controls (for TA97a and TA98, 20 μg/plate nitro phenylene diamine and for TA100, 1.5 μg/plate sodium azide) and negative controls were maintained concurrently for all the experiments. Three plates were used for each set. After 48 hours of incubation, the revertant colonies were counted. ANOVA test was performed at 0.05 level.

 

Amaranth [Food Red 2] was considered to be non - mutagenic under the study conditions.