Hydrogen bis[2-[[1-(3-chlorophenyl)-4,5-dihydro-3-methyl-5-oxo-1H-pyrazol-4-yl]azo]-5-sulphamoylbenzoato(2-)]chromate(1-)

Genetic toxicity: in vitro

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:

data from handbook or collection of data

Justification for type of information:

Data is from publication.

Data source

Reference

Reference Type:

publication

Title:

Gene mutation toxicity study of the test chemical

Author:

Seifried et al

Year:

2006

Bibliographic source:

Chem. Res. Toxicol.

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

as per Prival and Mitchell (1982)

Principles of method if other than guideline:

Salmonella/Mammalian-Microsome Mutagenicity Assay was conducted by using the given test chemical.

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Trisodium 5-hydroxy-1-(4-sulphophenyl)-4-(4-sulphophenylazo)pyrazole-3-carboxylate

Cas Number:

1934-21-0

Molecular formula:

C16H12N4O9-S2.3Na

IUPAC Name:

Trisodium 5-hydroxy-1-(4-sulphophenyl)-4-(4-sulphophenylazo)pyrazole-3-carboxylate

Details on test material:

IUPAC name: Trisodium 5-hydroxy-1-(4-sulphophenyl)-4-(4-sulphophenylazo)pyrazole-3-carboxylate
Commen name:Tartrazine
Molecular weight :534.3681 g/mol
Molecular formula:C16H12N4O9-S2.3Na
Smiles:n1(c2ccc(cc2)S(=O)(=O)[O-])c(c(c(n1)C(=O)[O-])/N=N/c1ccc(cc1)S(=O)(=O)[O-])O.[Na+].[Na+].[Na+]
InChI:1S/C16H12N4O9S2.3Na/c21-15-13(18-17-9-1-5-11(6-2-9)30(24,25)26)14(16(22)23)19-20(15)10-3-7-12(8-4-10)31(27,28)29;;;/h1-8,13H,(H,22,23)(H,24,25,26)(H,27,28,29);;;/q;3*+1/p-3/b18-17+;;
substance type: organic

Method

Target gene:

Histidine

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA98 and TA100

Details on mammalian cell type (if applicable):

not specified

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

Type and composition of metabolic activation system: cofactors (FMN, NADH, glucose-6-phosphate dehydrogenase, and glucose-6-phosphate), and test chemical were added, mixed, and incubated at 30 °C for 30 min without shaking.
- source of S9 : male uninduced hamster liver S9
- method of preparation of S9 mix: No data
- concentration or volume of S9 mix and S9 in the final culture medium: 30% v/v
- quality controls of S9 (e.g., enzymatic activity, sterility, metabolic capability) - Nitrogen was blown over the preincubation tube to keep the atmosphere reduced.

Test concentrations with justification for top dose:

0, 333, 1000, 3333, 6666, 10000 ug/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: Water
- Justification for choice of solvent/vehicle: Test chemical was soluble in water.

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

water

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

congo red

Details on test system and experimental conditions:

METHOD OF TREATMENT/ EXPOSURE:
- Cell density at seeding (if applicable): (1-2) x10^9 cells/mL
- Test substance added - preincubation

TREATMENT AND HARVEST SCHEDULE:
- Preincubation period, if applicable: 30 min
- Exposure duration/duration of treatment: 48 h
- Harvest time after the end of treatment (sampling/recovery times): No data

METHODS FOR MEASUREMENT OF CYTOTOXICITY
- Method.: Preincubation Method - All plates were counted with an Artek automated colony counter (Artek 880, DynaTech, Chantilly, VA) or Minicount colony counter (Imaging Products International, Inc., Chantilly, VA), which was calibrated prior to use.

Evaluation criteria:

All plates were observed for increase in mutant frequencies.

Statistics:

No data

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA98 and TA100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

True negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

RANGE-FINDING/SCREENING STUDIES (if applicable): The doses that were tested in the mutagenicity assay were selected based on the levels of cytotoxicity observed in a preliminary dose range-finding study using strain TA100. Ten dose levels of the chemical, one plate per dose, were tested in both the presence and the absence of induced hamster S9. If no toxicity was observed, a total maximum dose of 10 mg of test chemical per plate was used.

Remarks on result:

other: No mutagenic potential

Any other information on results incl. tables

Table

Test Chemical		Dose		TA98			TA100
			no S9	rat S9	Ham'r S9	no S9	rat S9	Ham'r S9
Prival Modification	Negative
		H2O	---	---	50 ± 5	---	---	217 ± 18
		333ug	---	---	53 ± 2	---	---	187 ± 28
		1000ug	---	---	55 ± 10	---	---	215 ± 14
		3333ug	---	---	52 ± 5	---	---	189 ± 6
		6666ug	---	---	59 ± 1	---	---	211 ± 50
		10000ug	---	---	57 ± 11	---	---	208 ± 20
		Positive 1	---	---	349 ± 33	---	---	785 ± 151
		Positive 2	---	---	397 ± 41	---	---	1316 ± 98

Applicant's summary and conclusion

Conclusions:

The given test chemical failed to induce mutation in Salmonella typhimurium TA98 and TA100 in the presence and absence of S9 metabolic activation and hence it is not likely to be mutagenic by in-vitro test.

Executive summary:

Bacterial Reverse Mutation Assay was conducted by using the given test chemical as per OECD Guideline 471 (Bacterial Reverse Mutation Assay) and Prival modification by Prival and Mitchell (1982). For tests using the FMN-modified assay, strains TA98 and TA100 were used at doses 0, 333, 1000, 3333, 6666, 10000 ug/plate dissolved in water. The bacteria, uninduced hamster liver S9 (30% v/v), cofactors (FMN, NADH, glucose-6-phosphate dehydrogenase, and glucose-6-phosphate), and test chemical was added, mixed, and incubated at 30 °C for 30 min without shaking. Nitrogen was blown over the preincubation tube to keep the atmosphere reduced. At the end of the incubation period, 2 mL of molten top agar was added to each sample tube and the mixture was poured on a minimal agar plate containing 0.5% glucose rather than the 2% glucose specified by Ames et al. The plates were then incubated at 37 °C for 48 h. The positive control in FMN experiment was Congo red. All plates were counted with an Artek automated colony counter or Minicount Colony counter, which was calibrated prior to use. The given test chemical failed to induce mutation in Salmonella typhimurium TA98 and TA100 in the presence and absence of S9 metabolic activation and hence it is not likely to be mutagenic by in-vitro test.