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Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Gene mutation in vitro:

Ames test:

Data available for the structurally and functonally similar read across chemicals was reviewed to determine the mutagenic nature of the test chemical Tetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]-4-hydroxy-5- [(1-oxopropyl)amino]naphthalene-2,7-disulphonate. The studies are as mentioned below:

Gene mutation toxicity study was performed to determine the mutagenic nature of the 60 -70% structurally and functionally similar 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.

Gene mutation toxicity study was performed to determine the mutagenic nature of the 60 -70% structurally and functionally similar test chemical. The study was performed as per the plate incorporation assay using Salmonella typhimurium strain TA98, TA1537, TA100, TA1535 with and without S9 metabolic activation system. The 2 ml of liquid top agar was cooled to 45°C and 0.1 ml of a broth cultureof microorganism and test substance in volumes of≤0.4 ml of DMSO was added prior to placing on minimal agar plates. The plates were incubated for 48 h at 37°C and the colonies which reverted to the prototroph were counted and compared to counts on the control plate (containing no test substance) to demonstrate mutagenicity or toxicity. Materials which caused a 2-fold increase of revertants, as compared to the number of spontaneous revertants on the control plates, were denoted as mutagens. Those which reduced the number of revertants were considered inhibitory. The test chemical did not result in a 2-fold increase in the number of revertants as compared to the number of spontaneous revertants on the control plates in 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.

Gene mutation toxicity study was also performed for another 60 -70% structurally and functoinally similar test chemical to evaluate its mutagenic nature. The study was performed as per the preincubation protocol using Salmonella typhimurium strain TA100, TA1535, TA1537, TA98 both in the presence and absence of S9 metabolic activation system at doses of 0, 100.0, 333.0, 1000.0, 3333.0 or 10000.0 µg/plate. Water was used as the vehicle. The plates were incubated for 48 hrs after 20 mins preincubation before the evaluation of the revertant colonies could be made. The test chemical did not induce mutation in the Salmonella typhimurium strain TA100, TA1535, TA1537, TA98 both in the presence and absence of S9 metabolic activation system and hence the chemical is not likely to classify as a gene mutant in vitro.

Based on the data available for the read across chemicals, the target chemical Tetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]-4-hydroxy-5- [(1-oxopropyl)amino]naphthalene-2,7-disulphonate is considered to not induce gene mutation is Salmonella typhimurium strains in the presence and absence of S9 metabolic activation system on the basis of the data available from the structurally and functionally similar read across chemical 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
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 and functionally similar read across chemicals
Reason / purpose:
read-across source
Related information:
Composition 1
Reason / purpose:
read-across source
Related information:
Composition 1
Reason / purpose:
read-across source
Related information:
Composition 1
Reference:
Composition 0
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
RA 1
Principles of method if other than guideline:
WoE derived based on the experimental data from structurally and functionally similar read across chemicals
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay
Test material information:
Composition 1
Target gene:
Histidine
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Remarks:
RA 1 / RA2
Details on mammalian cell lines (if applicable):
Not applicable
Additional strain characteristics:
other: frame-shift histidine mutants are TA1537 and TA98 and two base-pair substituted histidine mutants are TA1535 and TA100
Cytokinesis block (if used):
No data
Metabolic activation:
with and without
Metabolic activation system:
The microsomal fraction (S9) was prepared from male Sprague-Dawley rats
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Remarks:
RA 3
Details on mammalian cell lines (if applicable):
Not applicable
Additional strain characteristics:
not specified
Cytokinesis block (if used):
No data
Metabolic activation:
with and without
Metabolic activation system:
Male Sprague-Dawley rats and male Syrian hamsters were routinely used for the S9 preparation of the liver fractions
Test concentrations with justification for top dose:
1. 10-250 mg
2. 10-250 mg
3. 0, 100.0, 333.0, 1000.0, 3333.0, 10000.0 µg/plate
Vehicle:
1. - Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The chemical was soluble in DMSO

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

3. - Vehicle(s)/solvent(s) used: Water
- Justification for choice of solvent/vehicle: The test chemical was soluble in water
Negative controls:
not specified
Solvent controls:
not specified
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
other: Captan
Remarks:
RA 1
Negative controls:
not specified
Solvent controls:
yes
Remarks:
DMSO
True negative controls:
not specified
Positive controls:
not specified
Positive control substance:
not specified
Remarks:
RA 2
Negative controls:
not specified
Solvent controls:
yes
Remarks:
Water
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
9-aminoacridine
sodium azide
other: 4-nitro-o-phenylenediamine for TA98 (-S9); 2-aminoanthracene was used with all strains with hamster and rat liver metabolic activation systems.
Remarks:
RA 3
Details on test system and conditions:
1. 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 data

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

NUMBER OF REPLICATIONS: No data

NUMBER OF CELLS EVALUATED: No data

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

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

OTHER: No data

2. METHOD OF APPLICATION: In agar (plate incorporation)

DURATION
- Preincubation period: No data
- Exposure duration: 48 hrs
- Expression time (cells in growth medium): 48 hrs
- Selection time (if incubation with a selection agent): No data
- Fixation time (start of exposure up to fixation or harvest of cells): No data

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

NUMBER OF REPLICATIONS: No data

NUMBER OF CELLS EVALUATED: No data

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

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

OTHER: No data

3. METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: 20 mins
- Exposure duration: 48 hr
- Expression time (cells in growth medium): 48 hr
- Selection time (if incubation with a selection agent): No data
- Fixation time (start of exposure up to fixation or harvest of cells): No data

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

NUMBER OF REPLICATIONS: At least five dose levels of the chemicals were tested, with three plates per dose level.

NUMBER OF CELLS EVALUATED: No data

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

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

OTHER: No data


Rationale for test conditions:
1. No data
Evaluation criteria:
1. Mutagenicity was indicated by a clustering of revertant colonies directly around the test material or at the edge of the inhibitory zone.
2. The colonies which reverted to the prototroph were counted and compared to counts on the control plate (containing no test substance) to demonstrate mutagenicity or toxicity. Materials which caused a 2-fold increase of revertants, as compared to the number of spontaneous revertants on the control plates, were denoted as
mutagens. Those which reduced the number of revertants were considered inhibitory.
3. 1) mutagenic response: a dose-related, reproducible increase in the number of revertants over background, even if the increase was less than twofold;
2) nomutagenic response: when no increase in the number of revertants was elicited by the chemical;
3) questionable response: when there was an absence of a clear-cut dose-related increase in revertants; when the dose-related increases in the number of revertants were not reproducible; or when the response was of insufficient magnitude to support a determination of mutagenicity
Statistics:
1. No data
2. No data
3. Mean and Standard error of mean
Species / strain:
other: TA98, TA1537, TA100, TA1535
Remarks:
RA 1
Metabolic activation:
with and without
Genotoxicity:
not specified
Cytotoxicity:
not specified
Vehicle controls valid:
not specified
Negative controls valid:
not specified
Positive controls valid:
yes
Remarks on result:
other: No mutagenic potential
Species / strain:
other: TA98, TA1537, TA100, TA1535
Remarks:
RA2
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
not specified
Vehicle controls valid:
yes
Negative controls valid:
not specified
Positive controls valid:
not specified
Remarks on result:
other: No mutagenic potential
Species / strain:
other: TA100, TA1535, TA1537, TA98
Remarks:
RA 3
Metabolic activation:
with and 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:
1. No data
2. No data
3. TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: No data
- Effects of osmolality: No data
- Evaporation from medium: No data
- Water solubility: No data
- Precipitation: No data
- Other confounding effects: No data

RANGE-FINDING/SCREENING STUDIES: The chemical was initially tested with strain TA100 in the presence and the absence of the metabolic activation systems, over a wide dose range with an upper limit of 10 mg/plate, or less when solubility problems were encountered. Toxicity was evidenced by one or more of the following phenomena: appearance of his+ pinpoint colonies, reduced numbers of revertant colonies per plate, or thinning or absence of the bacterial lawn. Nontoxic chemicals were tested in the initial experiment up to the 10 mg/plate dose level, or to a level determined by their solubility. Toxic chemicals were tested up to a high dose which exhibited some degree of toxicity.

COMPARISON WITH HISTORICAL CONTROL DATA: No data

ADDITIONAL INFORMATION ON CYTOTOXICITY: No data
Conclusions:
The test chemical Tetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]-4-hydroxy-5- [(1-oxopropyl)amino]naphthalene-2,7-disulphonate is considered to not induce gene mutation is Salmonella typhimurium strains in the presence and absence of S9 metabolic activation system on the basis of the data available from the structurally and functionally similar read across chemical and hence is not likely to classify as a gene mutant in vitro.
Executive summary:

Data available for the structurally and functonally similar read across chemicals was reviewed to determine the mutagenic nature of the test chemical Tetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]-4-hydroxy-5- [(1-oxopropyl)amino]naphthalene-2,7-disulphonate. The studies are as mentioned below:

Gene mutation toxicity study was performed to determine the mutagenic nature of the 60 -70% structurally and functionally similar 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.

Gene mutation toxicity study was performed to determine the mutagenic nature of the 60 -70% structurally and functionally similar test chemical. The study was performed as per the plate incorporation assay using Salmonella typhimurium strain TA98, TA1537, TA100, TA1535 with and without S9 metabolic activation system. The 2 ml of liquid top agar was cooled to 45°C and 0.1 ml of a broth cultureof microorganism and test substance in volumes of≤0.4 ml of DMSO was added prior to placing on minimal agar plates. The plates were incubated for 48 h at 37°C and the colonies which reverted to the prototroph were counted and compared to counts on the control plate (containing no test substance) to demonstrate mutagenicity or toxicity. Materials which caused a 2-fold increase of revertants, as compared to the number of spontaneous revertants on the control plates, were denoted as mutagens. Those which reduced the number of revertants were considered inhibitory. The test chemical did not result in a 2-fold increase in the number of revertants as compared to the number of spontaneous revertants on the control plates in 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.

Gene mutation toxicity study was also performed for another 60 -70% structurally and functoinally similar test chemical to evaluate its mutagenic nature. The study was performed as per the preincubation protocol using Salmonella typhimurium strain TA100, TA1535, TA1537, TA98 both in the presence and absence of S9 metabolic activation system at doses of 0, 100.0, 333.0, 1000.0, 3333.0 or 10000.0 µg/plate. Water was used as the vehicle. The plates were incubated for 48 hrs after 20 mins preincubation before the evaluation of the revertant colonies could be made. The test chemical did not induce mutation in the Salmonella typhimurium strain TA100, TA1535, TA1537, TA98 both in the presence and absence of S9 metabolic activation system and hence the chemical is not likely to classify as a gene mutant in vitro.

Based on the data available for the read across chemicals, the target chemical Tetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]-4-hydroxy-5- [(1-oxopropyl)amino]naphthalene-2,7-disulphonate is considered to not induce gene mutation is Salmonella typhimurium strains in the presence and absence of S9 metabolic activation system on the basis of the data available from the structurally and functionally similar read across chemical 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:

Data available for the structurally and functonally similar read across chemicals was reviewed to determine the mutagenic nature of the test chemical Tetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]-4-hydroxy-5- [(1-oxopropyl)amino]naphthalene-2,7-disulphonate. The studies are as mentioned below:

Gene mutation toxicity study was performed to determine the mutagenic nature of the 60 -70% structurally and functionally similar 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.

Gene mutation toxicity study was performed to determine the mutagenic nature of the 60 -70% structurally and functionally similar test chemical. The study was performed as per the plate incorporation assay using Salmonella typhimurium strain TA98, TA1537, TA100, TA1535 with and without S9 metabolic activation system. The 2 ml of liquid top agar was cooled to 45°C and 0.1 ml of a broth cultureof microorganism and test substance in volumes of≤0.4 ml of DMSO was added prior to placing on minimal agar plates. The plates were incubated for 48 h at 37°C and the colonies which reverted to the prototroph were counted and compared to counts on the control plate (containing no test substance) to demonstrate mutagenicity or toxicity. Materials which caused a 2-fold increase of revertants, as compared to the number of spontaneous revertants on the control plates, were denoted as mutagens. Those which reduced the number of revertants were considered inhibitory. The test chemical did not result in a 2-fold increase in the number of revertants as compared to the number of spontaneous revertants on the control plates in 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.

Gene mutation toxicity study was also performed for another 60 -70% structurally and functoinally similar test chemical to evaluate its mutagenic nature. The study was performed as per the preincubation protocol using Salmonella typhimurium strain TA100, TA1535, TA1537, TA98 both in the presence and absence of S9 metabolic activation system at doses of 0, 100.0, 333.0, 1000.0, 3333.0 or 10000.0 µg/plate. Water was used as the vehicle. The plates were incubated for 48 hrs after 20 mins preincubation before the evaluation of the revertant colonies could be made. The test chemical did not induce mutation in the Salmonella typhimurium strain TA100, TA1535, TA1537, TA98 both in the presence and absence of S9 metabolic activation system and hence the chemical is not likely to classify as a gene mutant in vitro.

Based on the data available for the read across chemicals, the target chemical Tetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]-4-hydroxy-5- [(1-oxopropyl)amino]naphthalene-2,7-disulphonate (CAS no 80019 -42 -7) is considered to not induce gene mutation is Salmonella typhimurium strains in the presence and absence of S9 metabolic activation system on the basis of the data available from the structurally and functionally similar read across chemical and hence is not likely to classify as a gene mutant in vitro.

Justification for classification or non-classification

Based on the data available for the read across chemicals, the target chemical Tetrasodium 3-[[5-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]-2-sulphonatophenyl]azo]-4-hydroxy-5- [(1-oxopropyl)amino]naphthalene-2,7-disulphonate (CAS no 80019 -42 -7) is considered to not induce gene mutation is Salmonella typhimurium strains in the presence and absence of S9 metabolic activation system on the basis of the data available from the structurally and functionally similar read across chemical and hence is not likely to classify as a gene mutant in vitro.