Registration Dossier

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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study, to GLP.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2001
Report date:
2001

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:
other: 'Commission Directive 2000/32/EC, L1362000, Annexe 4D', dated May 19, 2000
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
Reaction mass of 6-ethyl-3-methyloct-5-en-1-ol and 6-ethyl-3-methyloct-6-en-1-ol
Molecular formula:
C11H22O
IUPAC Name:
Reaction mass of 6-ethyl-3-methyloct-5-en-1-ol and 6-ethyl-3-methyloct-6-en-1-ol
Test material form:
other: liquid
Details on test material:
Name: Super Muguet
Bathc Number: 9000408259
Aggregate state at Room Temperature: liquid
Colour: colourless to pale yellow
Stability in solvent: not indicated by the sponsor
Storage: room temperature
Expiry Date: 08 Dec 2002

Method

Target gene:
Histidine [S. typhimurium]
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
S9 liver microsomal fraction obtained from male rats, strain Wistar Hanlbm
Test concentrations with justification for top dose:
In the pre-experiment the concentration range of the test item was 3 - 5000 μg/plate. Due to toxic effects 1000 μg/plate were chosen as the maximal concentration for TA 98 and TA 100, and 333 1000 μg/plate for the remaining strains.

Strains TA 98 and TA 100: 3, 10, 33, 100, 333 and 1000 μg/plate
Strains TA 1535, TA 1537 and TA 102: 1, 3, 10, 33, 100 and 333 μg/plate
Vehicle / solvent:
Super Muguet was dissolved in DMSO, the solvent was chosen becuase of its solubility properties adn its relative non-toxicity to the bacteria.
Controls
Untreated negative controls:
yes
Remarks:
concurrent untreated and solvent controls were performed
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
sodium azide
methylmethanesulfonate
other: 2-aminoanthracene (2-AA); 4-nitro-o-phenylene-diamine (4-NOPD)
Details on test system and experimental conditions:
The test item Super Muguet was assessed for tis potential to induce gene mutations according to the plate incorporation test and the pre-incubation test using Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and TA 102.
The assay was performed in two independent experiments both with a dn without liver microsomal activation (S9). Each concentration and the controls, were tested in triplicate. The test item was tested at the following concentrations:
Strains TA 98 and TA 100: 3, 10, 33, 100, 333 and 1000 μg/plate
Strains TA 1535, TA 1537 and TA 102: 1, 3, 10, 33, 100 and 333 μg/plate
Evaluation criteria:
A test item is considered positive if a biologically relevant and dose related increase in the number of revertants is induced.
A test item producing neither a dose related increase in the numberof revertants nor a biologically relevant positive response at any one of the test points is considered non-mutagenic in this system.
A biologically relevant response is described as follows:
A test item is considered mitagenic if in strains TA 98, TA 100 and TA 102 the number of reversions will be at least twice as high and in strains TA 1535 and TA 1537 at least three times higher as compared to the spontaneous reversion rate (3, 4).
Also, a dose-dependant and reproducible increase in the number of revertants is regarded as an indication of possibly existing mutagenic potential of the test item regardless whether the highest dose will induce the above described enhamcement factors or not.
Statistics:
A statistical analysis of the data is not required.

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:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
Irregular background growth was observed at 333 μg/plate in strain TA 1545 (with and without S9) and in strain TA 1537 (with and without S9).
No substantial increase in revertant colony numbers of any five tester strains was observed following treatment with Super Muguet at any concentration level, neither in the presence nor absence of metabolic activation (S9 mix). There was also no tendency of higher mutation rates with increasing concentrations in the range below the generally acknowledged border of biological relevance.
The positive controls in the strains TA 1537 and TA 102 (with S9 mix) did not reach the required factors thrice (TA 1537) or twice (TA 102) of the corresponding solvent control. Therefore, the positive controkl of the parallel project 696700 was used for this study.
Appropriate reference mutagens were used as positive cintrols. They showed a distinct increase in induced revertant colonies.

Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

In conclusion, Super Muguet did not induce gene mutations by base pair changes or frameshifts in the genome of the strains used.
Therefore, Super Muguet is considered to be non-mutagenic in this Salmonella typhimurium reverse mutation assay.
Executive summary:

In a guideline study, conducted to GLP, Super Muguet was found to be non-mutagenic in the bacterial reverse mutation assay, tested in five strains of Salmonella typhimurium, in the presence and absence of metabolic activation.