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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data is from peer reviewed 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 tartrazine.
GLP compliance:
not specified
Type of assay:
bacterial gene mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
Details on test material
- Name of test material (as cited in study report): Tartrazine
- Molecular formula (if other than submission substance): C16-H12-N4-O9-S2.3Na.
C16-H9-N4-O9-S2.3Na
- Molecular weight (if other than submission substance): 534.3681 g/mol
- Substance type: Organic
- Physical state: No data available
Purity No data available
- Impurities (identity and concentrations): No data available
Specific details on test material used for the study:
- Name of test material: Tartrazine
- IUPAC name: Trisodium 5-hydroxy-1-(4-sulphophenyl)-4-(4-sulphophenylazo) pyrazole-3-carboxylate
- Molecular formula: C16H12N4O9S2.3Na
C16H9N4O9S23Na
- Molecular weight : 534.3681 g/mol
- Substance type: Organic
- Physical state: No data available
- Purity No data available
- Impurities (identity and concentrations): No data

Method

Target gene:
Histidine
Species / strain
Species / strain:
other: TA97a, TA98 and TA100
Details on mammalian cell lines (if applicable):
Not applicable
Additional strain characteristics:
not specified
Cytokinesis block (if used):
No data
Metabolic activation:
not specified
Metabolic activation system:
No data
Test concentrations with justification for top dose:
0, 100, 250, 500 or 1000 μg /plate
Vehicle:
- Vehicle(s)/solvent(s) used: Sterile double distilled water
- Justification for choice of solvent/vehicle: No data available
Controls
Negative controls:
not specified
Solvent controls:
yes
Remarks:
Sterile double distilled water
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
sodium azide
other: nitro phenylene diamine for TA97a and TA98
Details on test system and conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Preincubation period: No data available
- Exposure duration: 48 hrs
- Expression time (cells in growth medium): 48 hrs
- 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

SELECTION AGENT (mutation assays): No data available
SPINDLE INHIBITOR (cytogenetic assays): No data available
STAIN (for cytogenetic assays): No data available

NUMBER OF REPLICATIONS: Three plates were used for each set.

NUMBER OF CELLS EVALUATED: No data available

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data available

OTHER EXAMINATIONS:
- Determination of polyploidy: No data available
- Determination of endoreplication: No data available
- Other: No data available

OTHER: No data available
Rationale for test conditions:
No data
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:
other: TA97a, TA98 and TA100
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity:
not specified
Vehicle controls valid:
yes
Negative controls valid:
not specified
Positive controls valid:
yes
Remarks on result:
other: No mutagenic potential
Additional information on results:
No data

Any other information on results incl. tables

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

Dose

μG/plate

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

TA97a

TA98

TA100

100

126.70 ± 19.55

64.7 ± 4.04

110.7 ± 12.5

250

131.30 ± 18.50

60.0 ± 3.46

105.0 ± 15.13

500

118.00 ± 9.54

57.7 ± 21.2

125.0 ± 5.00

1000

113.30 ± 25.50

26.7 ± 3.21

115.0 ± 13.53

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

 

NPD= Nitrophynylenediamine

SA= Sodium azide

S.D.= Standard Deviation

Anova Value of TA97a (Tartrazine —1.256, ns)

TA 98 (Tartrazine –7.96*)

TA100 (Tartrazine—2.74, ns)

P< 0.01 (5.06)

Applicant's summary and conclusion

Conclusions:
Tartrazine did not induce gene mutation in Salmonella typhimurium strains TA97a, TA98 and TA100 and hence it is not likely to classify as a gene mutant in vitro.
Executive summary:

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

 

Tartrazine was dissolved in sterile double distilled water and was tested at a concentration of 0, 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.

Tartrazine did not induce gene mutation in Salmonella typhimurium strains TA97a, TA98 and TA100 and hence it is not likely to classify as a gene mutant in vitro.