4-[(2-chloro-4-nitrophenyl)azo]-N-ethyl-N-[2-[1-(2-methylpropoxy)ethoxy]ethyl]aniline

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Gene mutation in vitro:

Ames assay:

Gene mutation toxicity studies of the read across chemicals were reviewed to determine the mutagenic nature of the target chemical 4-((2-Chloro-4-nitrophenyl)azo)-N-ethyl-N-(2-(1-(2-methylpropoxy)ethoxy)ethyl)aniline. The studies are as mentioned below:

Ames assay was performed to determine the mutagenic nature of the given 50 -60% structurally and functionally similar test chemical. The study was performed using Salmonella typhimurium strains TA98 and TA100 in the presence and absence of metabolic activation system. The study was performed as per the princubation assay. The test chemical was preincubated with the bacterial strains for 1 hr and studied at dose levels of 0-100 µg/plate. The test chemical did not induce gene mutation in Salmonella typhimurium strains TA98 and TA100 in the presence and absence of metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.

In another study, bacterial gene mutation assay was performed to determine the mutagenic nature of another 50 -60% structurally and functionally similar test chemical. The study was performed using Salmonella typhimurium strains TA98, TA100, TA1535 and TA1537 in the presence and absence of S9 metabolic activation system. The study was performed as per the standard plate incorporation assay. The test chemical did not induce gene mutation in Salmonella typhimurium strains TA100, TA1535, TA1537 in the presence and absence of S9 metabolic activation system. It however induced gene mutation in Salmonella typhimurium strain TA98 in the presence of S9 metabolic activation system.

The given test chemical is therefore predicted to not induce gene mutation in Salmonella typhimurium strains in the presence and absence of metabolic activation system on the basis of the data available from the read across chemicals and hence it 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 similar read across chemicals

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: Refer below principle

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

Target gene:

Histidine

Species / strain / cell type:

S. typhimurium, other: TA98 and TA100

Remarks:

RA 1

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not specified

Species / strain / cell type:

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

Remarks:

RA 2

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:

10% (v/v) microsomal fractions (25% w/v) from Aroclor 1254-treated animals and supplemented with glucose 6-phosphate dehydrogenase (1 unit/plate).

Test concentrations with justification for top dose:

1. 0-100 µg/plate
2. No data

Vehicle / solvent:

1. No data
1. No data

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

not specified

True negative controls:

not specified

Positive controls:

not specified

Positive control substance:

not specified

Remarks:

RA 1

Details on test system and experimental conditions:

1. METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: 1 hr in shaking water bath at 37˚C
- 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: Triplicate

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: 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

Rationale for test conditions:

1. No data

Evaluation criteria:

1. The plates were observed for an increase in the number of revertants/plate
2. The plates were observed for an increase in the number of revertants/plate

Statistics:

1. Statistical analysis was carried out using Student's t-test.
2. No data

Species / strain:

S. typhimurium, other: TA98 and TA100

Remarks:

RA 1

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:

not specified

Species / strain:

S. typhimurium, other: TA100, TA1535, TA1537

Metabolic activation:

with and without

Genotoxicity:

not specified

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Additional information on results:

1. No data
2. No data

Remarks on result:

other: No mutagenic potential

Conclusions:

The given test chemical is predicted to not induce gene mutation in Salmonella typhimurium strains in the presence and absence of metabolic activation system on the basis of the data available from the read across chemicals and hence it is not likely to classify as a gene mutant in vitro.

Executive summary:

Gene mutation toxicity studies of the read across chemicals were reviewed to determine the mutagenic nature of the target chemical 4-((2-Chloro-4-nitrophenyl)azo)-N-ethyl-N-(2-(1-(2-methylpropoxy)ethoxy)ethyl)aniline. The studies are as mentioned below:

Ames assay was performed to determine the mutagenic nature of the given 50 -60% structurally and functionally similar test chemical. The study was performed using Salmonella typhimurium strains TA98 and TA100 in the presence and absence of metabolic activation system. The study was performed as per the princubation assay. The test chemical was preincubated with the bacterial strains for 1 hr and studied at dose levels of 0-100 µg/plate. The test chemical did not induce gene mutation in Salmonella typhimurium strains TA98 and TA100 in the presence and absence of metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.

In another study, bacterial gene mutation assay was performed to determine the mutagenic nature of another 50 -60% structurally and functionally similar test chemical. The study was performed using Salmonella typhimurium strains TA98, TA100, TA1535 and TA1537 in the presence and absence of S9 metabolic activation system. The study was performed as per the standard plate incorporation assay. The test chemical did not induce gene mutation in Salmonella typhimurium strains TA100, TA1535, TA1537 in the presence and absence of S9 metabolic activation system. It however induced gene mutation in Salmonella typhimurium strain TA98 in the presence of S9 metabolic activation system.

The given test chemical is therefore predicted to not induce gene mutation in Salmonella typhimurium strains in the presence and absence of metabolic activation system on the basis of the data available from the read across chemicals and hence it 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:

Gene mutation toxicity studies of the read across chemicals were reviewed to determine the mutagenic nature of the target chemical 4-((2-Chloro-4-nitrophenyl)azo)-N-ethyl-N-(2-(1-(2-methylpropoxy)ethoxy)ethyl)aniline. The studies are as mentioned below:

Ames assay was performed to determine the mutagenic nature of the given 50 -60% structurally and functionally similar test chemical. The study was performed using Salmonella typhimurium strains TA98 and TA100 in the presence and absence of metabolic activation system. The study was performed as per the princubation assay. The test chemical was preincubated with the bacterial strains for 1 hr and studied at dose levels of 0-100 µg/plate. The test chemical did not induce gene mutation in Salmonella typhimurium strains TA98 and TA100 in the presence and absence of metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.

In another study, bacterial gene mutation assay was performed to determine the mutagenic nature of another 50 -60% structurally and functionally similar test chemical. The study was performed using Salmonella typhimurium strains TA98, TA100, TA1535 and TA1537 in the presence and absence of S9 metabolic activation system. The study was performed as per the standard plate incorporation assay. The test chemical did not induce gene mutation in Salmonella typhimurium strains TA100, TA1535, TA1537 in the presence and absence of S9 metabolic activation system. It however induced gene mutation in Salmonella typhimurium strain TA98 in the presence of S9 metabolic activation system.

The given test chemical is therefore predicted to not induce gene mutation in Salmonella typhimurium strains in the presence and absence of metabolic activation system on the basis of the data available from the read across chemicals and hence it 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, 4-((2-Chloro-4-nitrophenyl)azo)-N-ethyl-N-(2-(1-(2-methylpropoxy)ethoxy)ethyl)aniline does not exhibit gene mutation in vitro. Hence the test chemical is not likely to be mutagenic as per the criteria mentioned in CLP regulation.