N,N-diethyl-p-(phenylazo)aniline

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Experimental study on genetic toxicity in vitro was conducted by Vincent F. Simmon (J Natl Cancer Inst ,1979) to determine the mutagenic nature of target substance4-(Diethylamino) azobenzene (2481-94-9). Genetic toxicity in vitro for 4-(Diethylamino) azobenzene was assessed for its possible mutagenic potential .For this purpose Salmonella-microsome assay was performed on Salmonella typhimurium strainTA1535, TA1537 and TA1538,using test substance concentration of 0-1000µg/plate (4.44µmol/plate).The test substance was exposed to the Salmonella typhimurium strainTA1535, TA1537 and TA1538 in the presence and absence of metabolic activation system.S9 mix, Adult male Sprague-Dawley rats (200- 250 g) pre-treated with Aroclor 1254 (500 mg/kg) were used to obtain liver for the metabolic activation system. The strains were checked periodically for relevant genetic markers and for rfa by crystal violet sensitivity.Each experiment included solvent controls as well as known direct-acting mutagens and a mutagen that required metabolic activation. No mutagenic effects were observed during the study. Therefore4-(Diethylamino) azobenzene was considered to be non mutagenic in Salmonella typhimurium strainTA1535, TA1537 and TA1538 by AMES assay. Hence it cannot be classified as gene mutant in vitro.

Link to relevant study records

Reference

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Justification for type of information:

Data is from publication .

Qualifier:

according to guideline

Guideline:

other: As mention below

Principles of method if other than guideline:

To evaluate the mutagenic potential of 4-(Diethylamino)azobenzene in Salmonella typhimurium strain TA1535, TA1537 and TA1538 by Salmonella-microsome assay.

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Specific details on test material used for the study:

- Name of test material: 4-(Diethylamino)azobenzene
- Molecular formula: C16H19N3
- Molecular weight: 253.347 g/mol
- Smiles notation: c1(N(CC)CC)ccc(\N=N\c2ccccc2)cc1
- InChl: 1S/C16H19N3/c1-3-19(4-2)16-12-10-15(11-13-16)18-17-14-8-6-5-7-9-14/h5-13H,3-4H2,1-2H3/b18-17+
- Substance type: Organic
- Physical state: Solid

Target gene:

Histidine

Species / strain / cell type:

S. typhimurium, other: strain TA1535, TA1537 and TA1538

Details on mammalian cell type (if applicable):

Not applicable.

Additional strain / cell type characteristics:

other: uvrB-deficient (to reduce the repair ofchemically induced lesions in the DNA) and are rfa (to prevent the synthesis of a normal lipopolysaccheride.

Cytokinesis block (if used):

not specified

Metabolic activation:

with and without

Metabolic activation system:

S9 mix, Adult male Sprague-Dawley rats (200- 250 g) pretreated with Aroclor 1254 (500 mg/kg) were used to obtain liver for the metabolic activation system

Test concentrations with justification for top dose:

0-1000µg/plate (4.44µ/mol)

Vehicle / solvent:

Yes ,but not specified .

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: detailed data not available.

Details on test system and experimental conditions:

Details on test system and conditions
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 24 hour


Other: The strains were checked periodically for relevant genetic markers and for rfa by crystal violet sensitivity.

Rationale for test conditions:

No data available.

Evaluation criteria:

The numbers of histidine-independent revertants for each S. typhimurium strain were taken from the linear portion of dose-response curves after the background was subtracted. The range of spontaneous revertants was 25-55 for TA1535 and 7-25 for TA1537. A positive response was defined as a reproducible, dose-related increase in the

Statistics:

Not specified.

Key result

Species / strain:

S. typhimurium, other: strain TA1535, TA1537 and TA1538

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:

Not specified.

Remarks on result:

other: No mutagenic effect were observed.

Conclusions:

4-(Diethylamino) azobenzene was evaluated for its mutagenic potential in Salmonella typhimurium strain TA1535, TA1537 and TA1538 by AMES assay.The test result was considered to be negative in the presece and absence of S9.

Executive summary:

Genetic toxicity in vitro for 4-(Diethylamino) azobenzene was assessed for its possible mutagenic potential .For this purpose Salmonella-microsome assay was performed on Salmonella typhimurium strainTA1535, TA1537 and TA1538,using test substance concentration of 0-1000µg/plate (4.44µmol/plate).The test substance was exposed to the Salmonella typhimurium strainTA1535, TA1537 and TA1538 in the presence and absence of metabolic activation system.S9 mix, Adult male Sprague-Dawley rats (200- 250 g) pre-treated with Aroclor 1254 (500 mg/kg) were used to obtain liver for the metabolic activation system. The strains were checked periodically for relevant genetic markers and for rfa by crystal violet sensitivity.Each experiment included solvent controls as well as known direct-acting mutagens and a mutagen that required metabolic activation. No mutagenic effects were observed during the study. Therefore4-(Diethylamino) azobenzene was considered to be non mutagenic in Salmonella typhimurium strainTA1535, TA1537 and TA1538 by AMES assay. Hence it cannot be classified as 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

Genotoxicity In-vitro

Various publications were reviewed to determine the mutagenic nature of 4-(Diethylamino) azobenzene (2481-94-9). The studies are as mentioned below:

Experimental study on genetic toxicity in vitro was conducted by Vincent F. Simmon (J Natl Cancer Inst ,1979) to determine the mutagenic nature of target substance4-(Diethylamino) azobenzene (2481-94-9). Genetic toxicity in vitro for 4-(Diethylamino) azobenzene was assessed for its possible mutagenic potential .For this purpose Salmonella-microsome assay was performed on Salmonella typhimurium strainTA1535, TA1537 and TA1538,using test substance concentration of 0-1000µg/plate (4.44µmol/plate).The test substance was exposed to the Salmonella typhimurium strainTA1535, TA1537 and TA1538 in the presence and absence of metabolic activation system.S9 mix, Adult male Sprague-Dawley rats (200- 250 g) pre-treated with Aroclor 1254 (500 mg/kg) were used to obtain liver for the metabolic activation system. The strains were checked periodically for relevant genetic markers and for rfa by crystal violet sensitivity.Each experiment included solvent controls as well as known direct-acting mutagens and a mutagen that required metabolic activation. No mutagenic effects were observed during the study. Therefore4-(Diethylamino) azobenzene was considered to be non mutagenic in Salmonella typhimurium strainTA1535, TA1537 and TA1538 by AMES assay. Hence it cannot be classified as gene mutant in vitro.

 Supporting Gene mutation toxicity study was performed by Yen-Ling Cheunget al.( CARCINOGENESIS , 1994) to determine the mutagenic nature of 4-(Diethylamino) azobenzene (2481-94-9) . Genetic toxicity study in vitro for 4-(Diethylamino) azobenzene (2481-94-9) was assessed for its possible mutagenic potential. For this purpose AMES assay was performed in S. typhimurium strains TA98, using test substance concentration of 0-100 µg/PLATE. A preincubation step was incorporated for 1h at 37°C in a shaking water bath. The test substance was exposed to salmonella typhimurium in the presence of metabolic activation. RAT, LIVER, MICROSOMES, AROCLOR 1254 and RAT, LIVER, MICROSOMES, 4-Diethylaminoazobenzene were used as Metabolic activation system. Evaluation was done considering a dose dependent increase in the number of histidine revertants/plate. Statistical analysis was carried out to observe the histidine revertants/plate. No mutagenic effects were observed during the study period in S. typhimurium strains TA98 in the presence of metabolic activation system. Therefore 4-(Diethylamino) azobenzene was considered to be non mutagenic in Ames test .Hence it cannot be classified as gene mutant in vitro.

Another supporting gene mutation toxicity study was performed by Joyce McCann et al (Food and Chemical Toxicology, 2002) to determine the mutagenic nature of4-(Diethylamino) azobenzene (2481-94-9). Genetic toxicity study in vitro for 4-(Diethylamino)azobenzene (2481-94-9)was assessed for its possible mutagenic potential. For this purpose AMES assay was performed in S. typhimurium strains TA98, using test substance concentration of <0.03µ mol /plate. A standard Salmonella plate test was performed. The test substance was exposed to salmonella typhimurium in the presence and absence of S9metabolic activation. S9 (RAT, LIVER, MICROSOMES, AROCLOR 1254) was used as Metabolic activation system. Evaluation was done considering a dose dependent increase in the number of histidine revertants/plate. Statistical analysis was carried out to observe the histidine revertants/plate. No mutagenic effects were observed during the study period in S. typhimurium strains TA98 in the presence and absence of metabolic activation system. Therefore 4-(Diethylamino) azobenzene was considered to be non mutagenic in Ames test .Hence it cannot be classified as gene mutant in vitro.

Based on the data available for the target chemical, it is concluded that 4-(Diethylamino) azobenzene (2481-94-9) does not exhibit gene mutation in vitro. Hence the test chemical is not likely to classify as a gene mutant in vitro.

Justification for classification or non-classification

Thus based on the above annotation and CLP criteria for the target chemical, it is concluded that 4-(Diethylamino) azobenzene (2481-94-9) does not exhibit gene mutation in vitro. Hence the test chemical is not likely to classify as a gene mutant in vitro.