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EC number: 236-740-8 | CAS number: 13472-08-7
Genetic toxicity: in vitro
Administrative data
- 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:
- other: see 'Remark'
- Remarks:
- This study is used for read-across and therefore has been assigned a reliability of 2 (reliable with restrictions). Otherwise the study has a reliability of 1 (reliable without restriction). This study was selected as the key study because the information provided for the hazard endpoint is sufficient for the purpose of classification and labeling and/or risk assessment. GLP Guideline study (OECD 471)
Data source
Referenceopen allclose all
- Reference Type:
- publication
- Title:
- Unnamed
- Year:
- 1 999
- Reference Type:
- other: english study summary
- Title:
- Unnamed
- Year:
- 1 999
- Reference Type:
- publication
- Title:
- OECD SIDS 2,2'-Azobis (2-methylpropionitrile) (CAS 78-67-1) - SIAM 9
- Author:
- Anonymous
- Year:
- 2 000
- Bibliographic source:
- UNEP Publications
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Qualifier:
- according to guideline
- Guideline:
- JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- 2,2'-dimethyl-2,2'-azodipropiononitrile
- EC Number:
- 201-132-3
- EC Name:
- 2,2'-dimethyl-2,2'-azodipropiononitrile
- Cas Number:
- 78-67-1
- Molecular formula:
- C8H12N4
- IUPAC Name:
- 2,2'-diazene-1,2-diylbis(2-methylpropanenitrile)
- Details on test material:
- - Name of test material (as cited in study report): 2,2'-AZOBIS(2-METHYLPROPIONITRILE)
- Physical state: white crystal
- Analytical purity: 99.9%
Constituent 1
Method
Species / strainopen allclose all
- Species / strain / cell type:
- S. typhimurium, other: TA 98, TA 100, TA 1535, TA 1537 and TA 97
- Additional strain / cell type characteristics:
- not applicable
- Species / strain / cell type:
- E. coli WP2 uvr A
- Additional strain / cell type characteristics:
- not applicable
- Metabolic activation:
- with and without
- Metabolic activation system:
- Rat S9 mix. Liver S9 homogenate was prepared from rats that have been induced with phenobarbital and 5,6-benzoflavone.
- Test concentrations with justification for top dose:
- 0, 313, 625, 1250, 2500 and 5000 µg/plate
- Vehicle / solvent:
- DMSO
Controls
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: (see below: details on test system and conditions)
- Details on test system and experimental conditions:
- METHOD OF APPLICATION:
- The test was performed according to the pre-incubation method.
NUMBER AND SELECTION OF DOSES, CONTROLS USED
- Five doses of test substance, together with the appropriate concurrent solvent and positive controls, were tested in triplicate on each tester strain with and without metabolic activation. The experiment was repeated twice except for TA1537 whic was tested four times.
-The dose-levels were the following: 0, 313,625, 1250, 2500 and 5000 µg/plate. All the strains were tested without S9 whereas TA97 was not part
of the experiment performed with S9.
The positive controls were as follows:
without S9 mix:
• sodium azide (NaN3): TA 1535 strain
• 9-Aminoacridine (9AA): TA 1537 and TA97 strains,
• 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide: TA 98 and TA 100 strains,
with S9 mix:
• 2-Aminoanthracene: TA 1535, TA 1537, TA 98 and TA 100 strains, - Evaluation criteria:
- Not precised
- Statistics:
- no
Results and discussion
Test resultsopen allclose all
- Species / strain:
- S. typhimurium, other: TA 98, TA 100, TA 1535, TA 1537 and TA 97
- Metabolic activation:
- without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Metabolic activation:
- with
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Species / strain:
- E. coli WP2 uvr A
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- See tables below.
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
Any other information on results incl. tables
Table 1 : Number of revertants per strain : 1st experiment
Table 2 : Number of revertants per strain : 2nd experiment
§Table 3 : Number of revertants per strain : Confirmation test
Table 4 : Number of revertants per strain: Additional experiment
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Applicant's summary and conclusion
- Conclusions:
- Interpretation of results: negative
This study and the conclusions which are drawn from it fulfill the quality criteria (validity, reliability, repeatability).
Under the experimental conditions described, the test substance 2,2'-AZOBIS(ISOBUTYRONITRILE) did not show any mutagenic activity in the bacterial reverse mutation test with Salmonella or E. coli. - Executive summary:
The potential of the test item 2,2-AZOBIS(ISOBUTYRONITRILE) to induce reverse mutation in Salmonella typhimurium was evaluated according to OECD 471 guideline in compliance with the Principles of Good Laboratory Practice.
Methods:
The test item was tested in two independent experiments, with and without a metabolic activation system, the S9 mix, prepared from a liver microsomal fraction (S9 fraction) of rats induced with phenobarbital and 5,6-benzoflavone. Both experiments were performed according to the preincubation method. Five strains of bacteria Salmonella typhimurium: TA 1535, TA 1537, TA 98, TA 100 and TA 97 were used except for TA 97 which was only used in the experiment without S9. E. coli WP2 uvrA was also tested with and without activation. Each strain was exposed to at least five dose-levels of the test item (three plates/dose-level). The evaluation of the toxicity was performed on the basis of the observation of the decrease in the number of revertant colonies and/or a thinning of the bacterial lawn. The test item 2,2-AZOBIS(ISOBUTYRONITRILE) was dissolved in dimethylsulfoxide (DMSO) and the following positive controls were used:
without S9 mix:
"sodium azide (NaN3): TA 1535 strain,
"9-Aminoacridine (9AA): TA 1537and TA 97 strains,
"2-(2-furyl)-3-(5-nitro-2-furyl) acrylamide : TA 98 and TA100 strains,
with S9 mix:
"2-Aminoanthracene: TA 1535, TA 1537, TA 98 and TA 100 strains
Results:
The total maximum dose of 5 mg per plate was selected as the highest dose of the experiment. The selected treatment-levels were 313, 625, 1250, 2500 and 5000 ug/plate for all strains with or without metabolic activation. The test item did not induce any significant increase in the number of revertants, in either experiment, in any of the five strains and no toxicity was observed.
Conclusion:
Under these experimental conditions, the test item 2,2-AZOBIS(ISOBUTYRONITRILE) did not show any mutagenic activity in the bacterial reverse mutation test with Salmonella typhimurium.
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