Trisodium 1-amino-4-[[3-[[4-chloro-6-[(sulphonatophenyl)amino]-1,3,5-triazin-2-yl]amino]-2,4,6-trimethyl-5-sulphonatophenyl]amino]-9,10-dihydro-9,10-dioxoanthracene-2-sulphonate

Genetic toxicity: in vitro

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:

Gene mutation toxicity study of the test chemical

Author:

Cameron et al

Year:

1987

Bibliographic source:

Mutation Research

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Principles of method if other than guideline:

Mutagenicity testing was performed by the standard plate-incorporation assay for the test chemical in Salmonella typhimurium by using TA1535, TA1537, TA1538, TA98 and TA100 strains.

GLP compliance:

not specified

Type of assay:

bacterial gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Disodium 4-hydroxy-3-[(4-sulphonatonaphthyl)azo]naphthalenesulphonate

EC Number:

222-657-4

EC Name:

Disodium 4-hydroxy-3-[(4-sulphonatonaphthyl)azo]naphthalenesulphonate

Cas Number:

3567-69-9

Molecular formula:

C20H14N2O7S2.2Na

IUPAC Name:

Disodium 4-hydroxy-3-[(4-sulphonatonaphthyl)azo]naphthalene sulphonate

Details on test material:

- Name of test material: Carmoisine- IUPAC name: disodium 4-hydroxy-3-[(4-sulfonato-1-naphthyl)diazenyl]naphthalene-1-sulfonate- Molecular formula: C20H14N2O7S2.2Na- Molecular weight: 502.4338 g/mol- Substance type: organic- Physical state: No data available - Purity 85% dye- Impurities (identity and concentrations): 15.0 %NaCI >0 1% subsidiary colors

Method

Target gene:

Histidine

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA1535, TA1537, TA1538, TA100 and TA98 were used.

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not specified

Cytokinesis block (if used):

No data

Metabolic activation:

with and without

Metabolic activation system:

S9 mix was prepared from Aroclor 1254-induced male Fischer 344 rats and Syrian golden hamsters.

Test concentrations with justification for top dose:

0, 333.0, 1000.0, 3333.0, 6666.0, 10000.0 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: The solvents used were water, dimethyl sulfoxide, and acetone (exact solvent details are not available)- Justification for choice of solvent/vehicle: Test chemical solubility in the solvents mentioned

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

other: 2-aminoanthracene (All strains ; with S9)

Details on test system and experimental conditions:

METHOD OF APPLICATION: plate incorporation.DURATION- Preincubation period: No data available- Exposure duration: No data available- Expression time (cells in growth medium): No data available- 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 availableSELECTION AGENT (mutation assays): No data availableSPINDLE INHIBITOR (cytogenetic assays): No data availableSTAIN (for cytogenetic assays): No data availableNUMBER OF REPLICATIONS: The chemical was tested at 5 dose levels in triplicate.NUMBER OF CELLS EVALUATED: No data availableDETERMINATION OF CYTOTOXICITY- Method: mitotic index; cloning efficiency; relative total growth; other: No data availableOTHER EXAMINATIONS:- Determination of polyploidy: No data available- Determination of endoreplication: No data available- Other: No data availableOTHER: No data available

Rationale for test conditions:

No data available

Evaluation criteria:

A response was considered positive if there was a dose-related increase in the number of revertants above spontaneous solvent controls with a 2-fold increase for strains TA1535, TA1538, TA98 and TA100, and a 3-fold increase for TA1537

Statistics:

No data available

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA1535, TA1537, TA1538, TA100 and TA98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS- Effects of pH: No data available- Effects of osmolality: No data available- Evaporation from medium: No data available- Water solubility: No data available- Precipitation: No data available- Other confounding effects: No data availableRANGE-FINDING/SCREENING STUDIES: The range of concentrations for testing was based on preliminary toxicity tests in which the viability of the bacterial cells on complete medium was measured at concentrations up to 10 mg/plate or to the limit of solubility. When solubility and toxicity were not limiting factors, the maximum concentration tested was 10 mg/plate.COMPARISON WITH HISTORICAL CONTROL DATA: No data availableADDITIONAL INFORMATION ON CYTOTOXICITY: No data available

Remarks on result:

other: No mutagenic potential

Any other information on results incl. tables

Table: Mutagenicity in Salmonella Assay

Dose (µg/plate)	Revertants/plate
	TA1535			TA1537			TA1538
	(-)S9	(R)S9	(H)S9	(-)S9	(R)S9	(H)S9	(-)S9	(R)S9	(H)S9
DMSO	43±6	30±4	28±2	7±5	11±2	10±3	11±3	23±7	20±6
Positive control	484±22	260±18	183±11	453±117	48±15	124±13	1042±47	155±2	793±18
333	38±3	31±6	30±3	10±1	9±3	12±2	10±2	24±4	26±1
1000	36±12	26±6	31±1	6±3	11±4	13±2	11±3	24±7	26±6
3333	31±8	24±3	18±6	7±2	6±1	12±4	10±4	15±4	16±1
6666	23±3	14±2	18±3	7±2	9±2	10±3	14±5	11±3	18±2
10000	29±5	20±11	17±5	9±1	6±2	9±1	13±1	16±2	23±3

 

Dose (µg/plate)	Revertants/plate
	TA98			TA100
	(-)S9	(R)S9	(H)S9	(-)S9	(R)S9	(H)S9
DMSO	13±1	25±4	27±6	151±7	149±16	177±17
Positive control	423±32	1745±72	1848±122	574±24	1079±108	1446±55
333	25±3	32±3	30±4	157±6	175±1	175±16
1000	18±1	33±10	38±3	152±12	183±5	176±11
3333	22±5	19±1	28±4	137±10	142±27	164±5
6666	25±4	27±8	28±10	150±21	125±9	133±7
10000	21±5	22±5	22±6	158±19	134±17	144± 7

Applicant's summary and conclusion

Conclusions:

The dye did not induce gene mutation in Salmonella typhimurium strains TA1535, TA1537, TA1538, TA100 and TA98 in the presence and absence of S9 metabolic activation system by plate-incorporation assay and hence is not likely to classify as a gene mutant in vitro.

Executive summary:

Bacterial gene mutation assay was performed for the test material in Salmonella typhimurium TA1535, TA1537, TA1538, TA100 and TA98 strains at a dose range of 0, 333.0, 1000.0, 3333.0, 6666.0 and 10000.0 µg/plate by Plate-incorporation method. Chemical was tested without metabolic activation and with S9 mix from Aroclor 1254-induced male Fischer 344 rats and Syrian golden hamsters. Appropriate positive and solvent controls were also incorporated in the study. Doses for the main study were based on the prelimicary study conducted. A response was considered positive if there was a dose-related increase in the number of revertants above spontaneous solvent controls with a 2-fold increase for strains TA1535, TA1538, TA98 and TA100, and a 3-fold increase for TA1537. The dye did not induce gene mutation in Salmonella typhimurium strains TA1535, TA1537, TA1538, TA100 and TA98 in the presence and absence of S9 metabolic activation system by plate-incorporation assay and hence is not likely to classify as a gene mutant in vitro.