Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Gene mutation in vitro:

Ames test:

The test chemical is not mutagenic to the Salmonella typhimurium strains in the presence and absence of liver S9 metabolic activation system 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
Justification for type of information:
Data is from data from handbook or collection of data
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Qualifier:
equivalent or similar to
Guideline:
other: As mention below
Principles of method if other than guideline:
WoE derived based on various experimental studies mention below
1,The test chemical was studied for its ability to induce mutations in strains of Salmonella typhimurium.
2,Gene mutation by the spot assay was performed for test chemical.
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:
1
Details on mammalian cell type (if applicable):
No data
Additional strain / cell type characteristics:
not specified
Species / strain / cell type:
S. typhimurium, other: TA100, TA98, TA1535, TA1537 and TA1538
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:
Liver S9 was prepared from Aroclor-induced male Fischer F344 rats
Test concentrations with justification for top dose:
1. 0, 222, 444, 888, 1776 or 3552 µg/plate
2. 200 µg/Plate
Vehicle / solvent:
1./2. - Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The test chemical is soluble in DMSO
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene
Remarks:
1
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
not specified
True negative controls:
yes
Positive controls:
yes
Positive control substance:
4-nitroquinoline-N-oxide
ethylmethanesulphonate
methylmethanesulfonate
other: Anthragallol, 2-Anthramine
Details on test system and experimental conditions:
1. METHOD OF APPLICATION: Liquid preincubation

DURATION
- Preincubation period: 30 mins in gyratory water bath
- Exposure duration: No data
- Expression time (cells in growth medium): No data
- 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 available

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

NUMBER OF REPLICATIONS: Triplicate

NUMBER OF CELLS EVALUATED: No data available

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

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

OTHER: No data available

2. METHOD OF APPLICATION: Spot test

DURATION
- Preincubation period: No data
- Exposure duration: 72 hrs
- Expression time (cells in growth medium): 72 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
Rationale for test conditions:
No data
Evaluation criteria:
1. The plates were observed for increase in the number of revertants/plate
2. The criteria adopted for scoring a mutagenic response in routine plate tests was that the observed number of revertants should exceed twice the background value for that given assay and exceed the 99.9% confidence limit based on our historical controls.
Statistics:
1. Mean ± SD
2. No data
Species / strain:
S. typhimurium, other: TA98 and TA100
Remarks:
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: TA100, TA98, TA1535, TA1537 and TA1538
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:
1/2.. No data
Remarks on result:
other: No mutagenic potential
Conclusions:
The test chemical is not mutagenic to the Salmonella typhimurium strains in the presence and absence of liver S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.
Executive summary:

Data available for the test chemical was reviewed to determine its mutagenic nature of test chemical. The studies are as mentioned below:

The test chemical was studied for its ability to induce mutations in strains of Salmonella typhimurium TA98 and TA100. The test compound was dissolved in DMSO and was tested at concentration of 0, 222, 444, 888, 1776 or 3552 µg/plate using Salmonella typhimurium TA100 and TA98 in the presence and absence of liver S9 metabolic activation system. Liquid preincubation assay was performed with a preicubation for 30 mins. The plates were observed for histidine independence. Concurrent solvent and positive controls were included in the study. The test chemical is not mutagenic to the Salmonella typhimurium TA100 and TA98 in the presence and absence of liver S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.

In another study, Gene mutation by the spot assay was performed for the test chemical. The given chemical was used at dose level of 200 µg/plate both with and without S9 metabolic activation system. The plates were incubated for 3 days at 37°C for the growth of colonies to occur. The plates were observed for an increase in the number of His+ revertants. The criteria adopted for scoring a mutagenic response in routine plate tests was that the observed number of revertants should exceed twice the background value for that given assay and exceed the 99.9% confidence limit based on our historical controls. Concurrent positive control was also included in the study. The test chemical did not induce mutation in Salmonella typhimurium strain TA100, TA98, TA1535, TA1537 and TA1538 with and without S9 metabolic activation system in the spot test performed and hence is not likely to classify for gene mutation in vitro.

Based on the data available for the test chemical is not mutagenic to the Salmonella typhimurium strains in the presence and absence of liver S9 metabolic activation system 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:

Data for the various test chemicals was reviewed to determine the mutagenic nature of 2,4-bis[(E)-2-(4-dodecylphenyl)diazen-1-yl]benzene-1,3-diol ( 65087-00-5). The studies are as mentioned below:

 

AMES Assay;

The test chemical was studied for its ability to induce mutations in strains of Salmonella typhimurium TA98 and TA100. The test compound was dissolved in DMSO and was tested at concentration of 0, 222, 444, 888, 1776 or 3552 µg/plate using Salmonella typhimurium TA100 and TA98 in the presence and absence of liver S9 metabolic activation system. Liquid preincubation assay was performed with a preicubation for 30 mins. The plates were observed for histidine independence. Concurrent solvent and positive controls were included in the study. The test chemical is not mutagenic to the Salmonella typhimurium TA100 and TA98 in the presence and absence of liver S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.

 

In another study, Gene mutation by the spot assay was performed for the test chemical. The given chemical was used at dose level of 200 µg/plate both with and without S9 metabolic activation system. The plates were incubated for 3 days at 37°C for the growth of colonies to occur. The plates were observed for an increase in the number of His+ revertants. The criteria adopted for scoring a mutagenic response in routine plate tests was that the observed number of revertants should exceed twice the background value for that given assay and exceed the 99.9% confidence limit based on our historical controls. Concurrent positive control was also included in the study. The test chemical did not induce mutation in Salmonella typhimurium strain TA100, TA98, TA1535, TA1537 and TA1538 with and without S9 metabolic activation system in the spot test performed and hence is not likely to classify for gene mutation in vitro.

Based on the data summarized, 2,4-bis[(E)-2-(4-dodecylphenyl)diazen-1-yl]benzene-1,3-diol ( 65087-00-5)did not induce gene mutation .Hence it is not likely to be mutagenic in vitro.

Justification for classification or non-classification

Based on the data available for the test chemical, 2,4-bis[(E)-2-(4-dodecylphenyl)diazen-1-yl]benzene-1,3-diol ( 65087-00-5) not likly to exhibit gene mutation in vitro. Hence the test chemical is not likely to be mutagenic as per the criteria mentioned in CLP regulation.