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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
September 30 - October 11, 1996
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1996

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Principles of method if other than guideline:
The study report is written in japanese language. The translated parts are presented in this summary.
GLP compliance:
no
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
1,1'-azobis(1-cyclohexanecarbonitrile)
EC Number:
218-254-8
EC Name:
1,1'-azobis(1-cyclohexanecarbonitrile)
Cas Number:
2094-98-6
Molecular formula:
C14H20N4
IUPAC Name:
1,1'-azobis(1-cyclohexanecarbonitrile)
Test material form:
solid
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: KCF9430
- Expiration date of the lot/batch: not stated
- Purity test date: not stated

RADIOLABELLING INFORMATION (if applicable)
not applicable

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Stable at room temperature, long term storage at 20 ° C or less
- Stability under test conditions: not stated
- Solubility and stability of the test substance in the solvent/vehicle: stated to be 0.1 g/100 g in water, 3 g/100 g DMSO, 23 g/100 g in Acetone
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: not stated

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
not stated

Method

Target gene:
his-, trp-
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Metabolic activation:
with and without
Metabolic activation system:
S9 mix
Test concentrations with justification for top dose:
1.2, 4.9, 20, 78, 313, 1250, 5000 µg/Plate (1st experiment)
313, 625, 1250, 2500, 5000 µg/Plate (2nd experiment)
Vehicle / solvent:
Acetone
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
sodium azide
benzo(a)pyrene
other: 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide, 2-Methoxy-6-chloro-9-[3-(2-chloroethyl)aminopropylamino]acridine *2HCl, 2-Aminoanthracene
Details on test system and experimental conditions:
two experiments performed:
METHOD OF APPLICATION: preincubation
- Cell density at seeding (if applicable): tested by Optical density (O.D.)

DURATION
- Preincubation period: 20 min
- Incubation duration: 48 h

NUMBER OF REPLICATIONS: 2
Rationale for test conditions:
not stated, but closely to OECD Guideline
Evaluation criteria:
A positive response is obtained
-if number of revertant colonies is twice as much as in negative control
- if a dose dependancy is observed.
Statistics:
none

Results and discussion

Test resultsopen allclose all
Key result
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
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:
valid
Positive controls validity:
valid

Applicant's summary and conclusion

Conclusions:
Under the described test conditions, the test substance did not induce a genotoxic response in bacteria with and without metablic activation.
Executive summary:

1,1'-azobis(1-cyclohexanecarbonitrile) was tested in a bacterial reverse mutation assay with and without metabolic activation.

The tested bacterial strains are S. typhimurium TA98, TA100, TA 1535, TA1537 and E. Coli WP2 uvrA. Two individual experiments were performed, both according to the pre-incubation method. Highest dose was 5000 µg/plate.

The number of revertant mutant colonies by treatment with the test substance in any strain did not increased more than twice as much in any concentration as compared with the solvent control. Additionally no dose response was observed.