[No public or meaningful name is available]

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:

from 2007-01-30 to 2007-02-07

Reliability:

1 (reliable without restriction)

Cross-reference

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2000/32/EC

Deviations:

no

Principles of method if other than guideline:

NA

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Details on test material:

Batch no: EZ06-B001;

Method

Target gene:

The Salmonella typhimurium histidine (his) reversion system measures his- => his+ reversions. The Salmonella typhimurium strains are constructed to differentiate between base pair (TA 1535, TA 100) and frameshift (TA 1537, TA 98) mutations. The Escherichia coli WP2 uvrA (trp) reversion system measures trp– => trp+ reversions. The Escherichia coli WP2 uvrA detect mutagens that cause other base-pair substitutions (AT to GC).

Species / strain

Species / strain 1

Species / strain / cell type:

E. coli WP2 uvr A

Species / strain 2

Species / strain / cell type:

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

Metabolic activation:

with and without

Metabolic activation system:

Male rodent activated microsomal fraction (S9-Mix)

Test concentrations with justification for top dose:

The test concentrations were: 5000; 1581.1; 500; 158.1; 50 and 15.81 µg/plate.

Vehicle / solvent:

DMSO

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

yes

Positive controls:

yes

Positive control substance:

other: different positive controls (see section "Any other information on materials and methods incl. tables" below)

Details on test system and experimental conditions:

NA

Evaluation criteria:

The colony numbers for control, positive control and the test plates were determined, the mean values and appropriate standard deviations were calculated.
The mutation factor (MF) was calculated by dividing the mean value of the revertant counts by the mean values of the solvent control (exact, not rounded values were used for this calculation).

Statistics:

NA

Results and discussion

Test results

Test results 1

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

Test results 2

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

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

In the experiment all of the observed revertant colony number increases were of minor intensity, not dose related, without any biological
significance (i.e. below the respective biological threshold value) and in the historical control range. Only for E. coli WP2 uvrA at 5000, 1581.1 and 158.1 and 50 mg/plate with S9 mix added were observed revertant colony numbers slightly above the historical control range with max. 66.3 revertants vs. 13 to 57.
Highest rates were observed in Salmonella typhimurium TA 1537, in experiment II at the concentration of 158.1 µg/plate, without metabolic
activation. The mutation factor value was: 1.57. There was no dose-dependent relationship and this value was well below the biological threshold
value.
In case of Salmonella typhimurium TA 98, TA 1535, TA 1537 and Escherichia coli WP2 uvrA slight decreases in revertant colony numbers
compared to the revertant colony numbers of the solvent control plates were detected in the performed experiments.
Because of the minor intensity of these variations (increases and decreases), the observed changes should be considered as reflecting the
biological variability of the test.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

NA

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative criteria: EU

AVE 5530 EVA-KET was non mutagenic in the bacterial reverse mutation assay.

Executive summary:

The reported data of this mutagenicity assay shows, that under the experimental conditions reported, the test item did not induce gene mutations by frameshift or base-pair substitution in the genome of the strains used. Therefore, AVE 5530 EVA-KET is considered non-mutagenic in this bacterial reverse mutation assay.