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

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Gene mutation in vitro:

Ames test:

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 is considered to not induce gene mutation in Salmonella typhimurium strains in the presence and absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.

Link to relevant study records

Reference

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

experimental data of read across substances

Justification for type of information:

Data for the target chemical is summarized based on the structurally similar read across chemicals

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Principles of method if other than guideline:

WoE derived based on the experimental data from struturally similar read across chemicals

GLP compliance:

not specified

Type of assay:

bacterial gene mutation assay

Target gene:

Histidine

Species / strain / cell type:

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

Remarks:

RA 1

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:

1. 0, 333.0, 1000.0, 3333.0, 6666.0, 10000.0 µg/plate2. 10-250 mg

Vehicle / solvent:

1.- 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 mentioned2. - Vehicle(s)/solvent(s) used: DMSO- Justification for choice of solvent/vehicle: The chemical was soluble in DMSO

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)

Remarks:

1

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

not specified

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: Captan

Remarks:

2

Details on test system and experimental conditions:

1. 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 2. METHOD OF APPLICATION: In agar (Spot test)DURATION- Preincubation period: No data- Exposure duration: No data- Expression time (cells in growth medium): No data- Selection time (if incubation with a selection agent): No data- Fixation time (start of exposure up to fixation or harvest of cells): No dataSELECTION AGENT (mutation assays): No dataSPINDLE INHIBITOR (cytogenetic assays): No dataSTAIN (for cytogenetic assays): No dataNUMBER OF REPLICATIONS: No dataNUMBER OF CELLS EVALUATED: No dataDETERMINATION OF CYTOTOXICITY- Method: mitotic index; cloning efficiency; relative total growth; other: No dataOTHER EXAMINATIONS:- Determination of polyploidy: No data- Determination of endoreplication: No data- Other: No dataOTHER: No data

Rationale for test conditions:

No data available

Evaluation criteria:

1. 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 TA15372. Mutagenicity was indicated by a clustering of revertant colonies directly around the test material or at the edge of the inhibitory zone.

Statistics:

No data available

Species / strain:

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

Remarks:

RA 1

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

Species / strain:

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

Remarks:

RA 2

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

1. 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 available2. No data

Remarks on result:

other: No mutagenic potential

Conclusions:

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 is predicted to not induce gene mutation in Salmonella typhimurium strains in the presence and absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.

Executive summary:

Various studies availave for the test chemical were reviewed to determine the mutagenic nature of Trisodium1-amino-4-[[3-[[4-chloro-6-[(sulphonato phenyl)amino]-1,3,5-triazin-2-yl] amino] -2,4,6-trimethyl-5-sulphonatophenyl] amino]-9,10-dihydro-9,10-dioxoanthracene-2-sulphonate. The studies are as mentioned below:

Study 1:

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.

Study 2:

Gene mutation toxicity study was also performed to determine the mutagenic nature of the test chemical. Spot test was performed at dose levels from 10-250 mg using Salmonella typhimurium strain TA98, TA1537, TA100, TA1535 with and without S9 metabolic activation system. Captan was used as positive control chemical and the solvent control used was DMSO. Mutagenicity was indicated by a clustering of revertant colonies directly around the test material or at the edge of the inhibitory zone. The test chemical did not induce clustering of revertant colonies directly around the test material or at the edge of the inhibitory zone using Salmonella typhimurium strain TA98, TA1537, TA100, TA1535 in the presence and absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.

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 is considered to not induce gene mutation in Salmonella typhimurium strains in the presence and absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Gene mutation in vitro:

Various studies available for the test chemical were reviewed to determine the mutagenic nature of Trisodium1-amino-4-[[3-[[4-chloro-6-[(sulphonato phenyl)amino]-1,3,5-triazin-2-yl] amino] -2,4,6-trimethyl-5-sulphonatophenyl] amino]-9,10-dihydro-9,10-dioxoanthracene-2-sulphonate (CAs no 72214 -18 -7). The studies are as mentioned below:

Study 1:

Bacterial gene mutation assay was performed for the test material in Salmonella typhimuriumTA1535, 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.

Study 2:

Gene mutation toxicity study was also performed to determine the mutagenic nature of the test chemical. Spot test was performed at dose levels from 10-250 mg using Salmonella typhimurium strain TA98, TA1537, TA100, TA1535 with and without S9 metabolic activation system. Captan was used as positive control chemical and the solvent control used was DMSO. Mutagenicity was indicated by a clustering of revertant colonies directly around the test material or at the edge of the inhibitory zone. The test chemical did not induce clustering of revertant colonies directly around the test material or at the edge of the inhibitory zone using Salmonella typhimurium strain TA98, TA1537, TA100, TA1535 in the presence and absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.

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 (72214 -18 -7) is considered to not induce gene mutation in Salmonella typhimurium strains in the presence and absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.

Justification for classification or non-classification

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 (72214 -18 -7) is considered to not induce gene mutation in Salmonella typhimurium strains in the presence and absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.