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Toxicological information

Genetic toxicity: in vitro

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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:
from 09 MAR 1999 to 18 JUN 1999
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study (according to OECD 471).

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1999
Report date:
1999

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
Deviations:
not specified
GLP compliance:
yes
Remarks:
in compliance with US EPA FIFRA (40CFR Part 160) and/or EPA TSCA (49CFR 792) Good Laboratory Practice Standards
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
Perhydroazepine
EC Number:
203-875-9
EC Name:
Perhydroazepine
Cas Number:
111-49-9
Molecular formula:
C6H13N
IUPAC Name:
azepane
Details on test material:
- Name of test material (as cited in study report): Hexamethyleneimine (HMI)
- Substance type: colorless liquid
- Physical state: fluid
- Analytical purity: 99.5 %
- Impurities (identity and concentrations): water
- Stability under test conditions: the test substance appeared to be stable under the conditions of the study, no evidence of instability was observed.

Method

Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535
Additional strain / cell type characteristics:
not specified
Species / strain / cell type:
S. typhimurium TA 100
Additional strain / cell type characteristics:
not specified
Species / strain / cell type:
S. typhimurium TA 98
Additional strain / cell type characteristics:
not specified
Species / strain / cell type:
other: S. typhimurium TA97a
Additional strain / cell type characteristics:
not specified
Species / strain / cell type:
E. coli WP2 uvr A
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
rat liver S9 mix
Test concentrations with justification for top dose:
0, 10, 50, 100, 500, 1000, 2500, 5000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: sterile water
Controlsopen allclose all
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: TA 97a: 9,10-Dimethyl-1,2-benzanthracene; TA 98, TA 100, TA 1535, E.coli WP2: 2-Aminoanthracene
Remarks:
with metabolic activation
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: TA 97a: ICR 191 Acridine mutagen; TA 98: 2NF; TA 100, TA 1535: Sodium azide; E.coli WP2: N-Ethyl-N-nitro-N-nitroguanidine
Remarks:
without metabolic activation
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY
- Method: reduction of the microcolony background lawn and concentration-related reduction in the mean number of revertants per plate
Evaluation criteria:
A test substance was classified as positive (i.e. mutagenic) if the mean number of revertants in any strain at any test substance concentrationw as at least 2 times greater than the mean of concurrent vehicle control and there was a concentration related increase in the mean revertants per plate in the same strain.
A test substance was classified as negative (i.e. not mutagenic) if there were no test substance concentrations with a mean number of revertants that was at least 2 times greater than the mean of concurrent vehicle control and there was no concentration related increase in the mean revertants per plate in that same strain.
Results not meerinig criteria for positive or negative classification were evaluated using scientific judgment and experience and may have been reported equivocal.
Statistics:
Data for each tester strain were evaluated independently. For each tester strain, the mean number of revertants and the standard deviation at each concentration in the presence and absence of exogenous metabolic activation system were calculated.

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
starting at concentration of 1000 µg/plate and higher (without metabolic activation) or in the highest concentration (with metabolic activation)
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
starting at concentration of 1000 µg/plate and higher (without metabolic activation) or in the two highest concentrations (with metabolic activation)
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
starting at concentration of 1000 µg/plate and higher (without metabolic activation) or in the two highest concentrations (with metabolic activation)
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Species / strain:
other: S. typhimurium TA97a
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
starting at concentration of 1000 µg/plate and higher (without metabolic activation) or in the two highest concentrations (with metabolic activation)
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
starting at concentration of 1000 µg/plate and higher (without metabolic activation) or in the two highest concentrations (with metabolic activation)
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: no precpipitation occurred

COMPARISON WITH HISTORICAL CONTROL DATA: the mean number of revertants scored for the solvent control of each strain was within the laboratory´s historical data range.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

The appropriate reference mutagenes for each tester strain showed distinct positive mutagenic effects.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

The test item showed no mutagenic activity in a plate incorporation assay with and without metabolic activation.
Executive summary:

Mutagenic activity of the test item was investigated in Salmonella typhimurium strains TA 1535, TA 97a, TA98 and TA100 as well as Escherichia coli strain WP2 uvrA with (induced rat liver S9 mix) and without metabolic activation at concentrations of 0, 10, 50, 100, 500, 1000, 2500 and 5000 µg/plate using the plate incorporation assay.

The test item did not reveal any mutagenic activity under the conditions tested. The appropriate reference mutagenes showed distinct positive mutagenic effects.