2H-Azepin-2-one, hexahydro-, polymer with 1,6-diisocyanatohexane

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2007-08-15 to 2007-08-28

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Principles of method if other than guideline:

Remark: Both TA 102 and E.coli WP2 were not tested (not required by applied version of guideline).

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Copolymer of hexahydro-2H-azepin-2-one and 1,6-diisocyanatohexane

EC Number:

607-997-7

Cas Number:

26776-30-7

Molecular formula:

Exact identification is not feasible

IUPAC Name:

Copolymer of hexahydro-2H-azepin-2-one and 1,6-diisocyanatohexane

Test material form:

other: viscous liquid

Details on test material:

- Appearance: clear, colorless
- Physical state: viscous liquid

Method

Target gene:

mutated gene loci responsible for histidine auxotrophy

Species / strain

Species / strain / cell type:

other: Salmonella typhimurium TA 98, TA100, TA 102, TA 1535, TA 1537

Additional strain / cell type characteristics:

other: histidine auxotroph

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254-induced rat liver S9 fraction, prepared from  male Sprague Dawley rats

Test concentrations with justification for top dose:

20 to 5000 µg/plate

Vehicle / solvent:

dimethyl sulfoxide,  DMSO (CAS No. 67-68-5)

Controls

Untreated negative controls:

yes

Remarks:

untreated

Negative solvent / vehicle controls:

yes

Remarks:

Ethylen glycol dimethylether (EGDE)

True negative controls:

no

Positive controls:

yes

Remarks:

details see below

Positive control substance:

other: without metabolic activation: 4-Nitro-1,2-phenylene (TA 98; TA 1537), Nitrofurantoin (TA100), Sodium azide (TA 1535), Mitomycin C (TA 102); with metabolic activation: Aminoanthracene

Details on test system and experimental conditions:

Ames test
SYSTEM OF TESTING
- Metabolic activation system:  Aroclor 1254-induced rat liver S9 fraction, prepared from  male Sprague Dawley rats
ADMINISTRATION: 
- Dosing:    
Plate incorporation test: 50/158/500/1581/5000 µg/plate (+/- metabolic  activation)   
- Number of replicates: 2
- Repeat: preincubation
- Application: solvent ethylen glycol dimethylether (EGDE)
- Positive and negative control groups and treatment:    
positive without metabolic activation:   
TA 98: 4-Nitro-1,2-phenylene diamine (0.5 µg/plate in dimethyl sulfoxide)   
TA 100: Nitrofurantoin (0.2 µg/plate in dimethyl sulfoxide)   
TA 102: Mitomycin C (0.2 µg/platein deionized water)
TA 102: Cumene hydroperoxide (50 µg/plate in dimethyl sulfoxide) in plate preincubtion trials
TA 1535: Sodium azide (10 µg/plate in dimethyl sulfoxide)   
TA 1537: 4-Nitro-1,2-phenylene diamine (10 µg/plate in dimethyl sulfoxide)   
positive with metabolic activation and activity of metabolic system:   all strains: 2-aminoanthracene (2.5 µg/plate in dimethyl sulfoxide)   
negative: solvent control, 100 µl/plate (pre-incubation: 50 µl/plate)
- Pre-incubation: 20 minutes at 37 °C   incubation 48 hours at 37 °C

Evaluation criteria:

CRITERIA FOR EVALUATING RESULTS:    
- negative controls and to be within the expected range, as defined by published data
- positive controls had to show sufficient effects, as defined by the laboratories experience
- titer determinations had to demonstrate sufficient bacterial density in the suspension

Results and discussion

Test results

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Remarks:

doses up to and including 5000 µg per plate did not cause any bacteriotoxic effects

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

GENOTOXIC EFFECTS: 
- With metabolic activation: None
- Without metabolic activation: None   
The positive controls w ere functional.
PRECIPITATION CONCENTRATION: Precipitation occurred at the dose 1581 µ per plate and above
CYTOTOXIC CONCENTRATION (including effects on background lawn): 
- Doses up to and including 5000 µg per plate did not cause any bacteriotoxic effects

Remarks on result:

other: other:

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

no other results

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

The test item was considered to be non-mutagenic without and with S9 mix in the plate incorporation as well as in the preincubation modification of the Salmonella/microsome test.

Executive summary:

The test item was tested for its ability to induce reverse mutations in an in vitro bacterial system.

Salmonella typhimurium strains TA 98, TA 100, TA 102, TA 1535 and TA 1537 were treated with 2H-Azepin-2-one, hexahydro-, polymer with 1,6-diisocyanatohexane by the Ames test plate incorporation as well as the preincubation method. Five dose levels covering the range between 50 and 5000 µg/plate, in triplicate both with and without the addition of a metabolizing system were employed.

All five bacterial strains exhibited mutagenic response to the appropriate positive control substances. Solvent controls were also tested with each strain and the mean numbers of spontaneous revertants were in an acceptable range.

Mutagenic activity of the test compound to one or more of the tester strains was not observed in either experiment with and without metabolic activation. Therefore, the test item was considered to be non-mutagenic without and with S9 mix in the plate incorporation as well as in the preincubation modification of the Salmonella/microsome test.