Bromo(hexahydro-2H-azepin-2-onato-N)magnesium

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

01/2008 to 05/2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Bromo(hexahydro-2H-azepin-2-onato-N)magnesium

EC Number:

241-158-2

EC Name:

Bromo(hexahydro-2H-azepin-2-onato-N)magnesium

Cas Number:

17091-31-5

Molecular formula:

C6H10BrMgNO

IUPAC Name:

magnesium(2+) ion 2-oxoazepan-1-ide bromide

additive 1

Reference substance name:

ε-caprolactam

EC Number:

203-313-2

EC Name:

ε-caprolactam

Cas Number:

105-60-2

Molecular formula:

C6H11NO

IUPAC Name:

azepan-2-one

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 07072601
- Expiration date of the lot/batch: 26.07.2008

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature 20 ± 5 °C

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: no

Method

Species / strain

Species / strain / cell type:

other: TA 97a, TA 98, TA 100, TA 102 and TA 1535

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

S9

Test concentrations with justification for top dose:

4986 to 50 μg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: water

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

yes

Positive controls:

yes

Positive control substance:

sodium azide

benzo(a)pyrene

other: 4-Nitro-1,2-phenylene diamine (NPD) in DMSO, 2-Amino anthracene (2-AA) in DMSO

Details on test system and experimental conditions:

Plate incorporation method

Per strain and dose, four plates with and four plates without S9 mix were used. 10 ml of
the test solution of the appropriate concentration were membrane filtrated into sterile vessels.
Top agar basis was melted in a microwave oven, after melting, 10 ml of histidinebiotin-
solution 0.5 mMol per 100 ml basis was added and the bottle was placed in the
water bath at 45 °C.
0.1 ml of the appropriate solution of the test item were given into a sterile tube. After mixing
with 0.1 ml overnight culture of the respective strain and 0.5 ml phosphate buffer
(only for treatments without S9) or 0.5 ml S9 mix, 2 ml Top-Agar were added. The mixture
was gently vortexed, then poured on a minimal glucose plate and distributed evenly,
using a Drigalski spatula. The plates were closed, covered with brown paper and left to
harden for a few minutes, then inverted and placed in the dark incubator at 37 °C.

Pre-incubation method

Per strain and dose, four plates with and four plates without S9 mix were used. 10 ml of
the test solution of the appropriate concentration were membrane filtrated into sterile vessels.
Top agar basis was melted in a microwave oven, after melting, 10 ml of histidinebiotin-
solution 0.5 mMol per 100 ml basis was added and the bottle was placed in the
water bath at 45 °C.
0.1 ml of the appropriate solution of the test item were given into a sterile tube. After mixing
with 0.1 ml overnight culture of the respective strain, 0.5 ml phosphate buffer (only
for treatments without S9) or 0.5 ml S9 mix were added. The mixture was incubated in an
incubation chamber at 37°C for 20 minutes. During this time the vessels were aerated
through careful shaking. Then 2 ml top agar were added. The mixture was vortexed gently,
then poured on a minimal glucose plate and distributed evenly, using a Drigalski spatula.
The plates were closed, covered with brown paper and left to harden for a few minutes,
then inverted and placed in the dark incubator at 37 °C.

Evaluation criteria:

The colonies were counted visually, the numbers were recorded.

Statistics:

A spreadsheet software (Microsoft Excel®) was used to calculate mean values and standard deviations as well as the increase factor of revertant induction.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 97

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Test results 2

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Test results 3

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Test results 4

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Test results 5

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Applicant's summary and conclusion

Conclusions:

Under the conditions of the test, the test item didn't show mutagenic effects towards Salmonella typhimurium, strains TA 97a, TA 98, TA 100, TA 102 and TA 1535. Therefore, no concentration-effect relationship could be determined.

Executive summary:

The test item Bromo(hexahydro-2H-azepin-2-onato-N)magnesium is considered as "not mutagenic under the conditions of the test".