Registration Dossier

Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report Date:
2017

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
GLP compliance:
yes
Type of assay:
bacterial forward mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Specific details on test material used for the study:
Test material name (as stated in the report): CERVOLIDE
Batch No.: VE00445215
Expiry date: 07 June 2018

Method

Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Metabolic activation:
with and without
Test concentrations with justification for top dose:
CERVOLIDE was tested in the tester strains TA1535, TA1537, TA98, TA100 and WP2uvrA
with concentrations of 5.4, 17, 52, 164, 512, 1600 and 5000 μg/plate in the absence and
presence of 5% (v/v) S9-mix.
Vehicle / solvent:
Saline and DMSO
Controlsopen allclose all
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
sodium azide
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
2-nitrofluorene
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
methylmethanesulfonate
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
4-nitroquinoline-N-oxide
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: ICR-191

Results and discussion

Test results
Key result
Species / strain:
other: TA1535, TA1537, TA98, TA100 and WP2uvrA
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

Any other information on results incl. tables

All bacterial strains showed negative responses over the entire dose-range, i.e. no significant

dose-related increase in the number of revertants in two independently repeated experiments.

The negative control values were within the laboratory historical control data ranges.

The strain-specific positive control values were within the laboratory historical control data

ranges indicating that the test conditions were adequate and that the metabolic activation

system functioned properly, except the responses for TA1535 (presence of S9-mix) and

TA1537 (absence of S9-mix) in the second experiment. The purpose of the positive control is

as a reference for the test system, where a positive response is required to check if the test

system functions correctly. Since the values were more than 10 and 342 times greater than

the concurrent solvent control values for the tester strains TA1535 and TA1537, respectively,

these deviations in the mean plate counts of the positive controls had no effect on the results

of the study.

Applicant's summary and conclusion

Conclusions:
In conclusion, based on the results of this study it is concluded that CERVOLIDE is not
mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli
reverse mutation assay.
Executive summary:

All bacterial strains showed negative responses over the entire dose-range, i.e. no significant

dose-related increase in the number of revertants in two independently repeated experiments.

The negative control values were within the laboratory historical control data ranges.

The strain-specific positive control values were within the laboratory historical control data

ranges indicating that the test conditions were adequate and that the metabolic activation

system functioned properly, except the responses for TA1535 (presence of S9-mix) and

TA1537 (absence of S9-mix) in the second experiment. The purpose of the positive control is

as a reference for the test system, where a positive response is required to check if the test

system functions correctly. Since the values were more than 10 and 342 times greater than

the concurrent solvent control values for the tester strains TA1535 and TA1537, respectively,

these deviations in the mean plate counts of the positive controls had no effect on the results

of the study.

In conclusion, based on the results of this study it is concluded that CERVOLIDE is not

mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli

reverse mutation assay.