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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
January 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report Date:
2018

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
other: liquid
Details on test material:
Identification: Bourgeonal
CAS number: 18127-01-0
Description: Clear colourless liquid
Storage conditions: At ambient temperature under nitrogen.
Batch number: VE000010051
Date of receipt: 20 November 2008
Expiry date: 4 August 2009
Purity: 98.4%

Method

Target gene:
Histidine for Salmonella.
Tryptophan for E.Coli
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
S9
Test concentrations with justification for top dose:
Based on the results of the first mutation experiment, BOURGEONAL was tested up to concentrations of 512 and 5000 μg/plate in the absence and presence of 10% (v/v) S9-mix in the Salmonella typhimurium and Escherichia coli strain, respectively.
Vehicle / solvent:
DMSO
Controls
Untreated negative controls:
yes
Remarks:
DMSO
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
Positive controls:
yes
Positive control substance:
4-nitroquinoline-N-oxide
2-nitrofluorene
sodium azide
methylmethanesulfonate
other: ICR-191; 2-aminoanthracene
Details on test system and experimental conditions:
According to OECD 471
Rationale for test conditions:
According to OECD 471
Evaluation criteria:
According to OECD 471
Statistics:
According to OECD 471

Results and discussion

Test resultsopen allclose all
Key result
Species / strain:
S. typhimurium TA 1535
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
Key result
Species / strain:
S. typhimurium TA 1537
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
Key result
Species / strain:
S. typhimurium TA 98
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
Key result
Species / strain:
S. typhimurium 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
Species / strain:
E. coli WP2 uvr A
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

Applicant's summary and conclusion

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

The objective of this study was to determine the potential of BOURGEONAL and/or its

metabolites to induce reverse mutations at the histidine locus in several strains of Salmonella

typhimurium (S. typhimurium; TA98, TA100, TA1535, and TA1537), and at the tryptophan

locus of Escherichia coli (E. coli) strain WP2uvrA in the presence or absence of an exogenous

mammalian metabolic activation system (S9).

The study procedures described in this report were based on the most recent OECD, EC and

METI guidelines.

Batch SC00021197 of BOURGEONAL was a pale yellow liquid with a purity of 97.2%. The

test item was dissolved in dimethyl sulfoxide.

In the first mutation assay, the test item was tested up to concentrations of 5000 μg/plate in

the absence and presence of 5% (v/v) S9-mix. The test item precipitated on the plates at this

dose level. Cytotoxicity, as evidenced by a decrease in the number of revertants, reduction of

the bacterial background lawn and/or the presence of microcolonies, was observed in all tester

strains in the absence and presence of S9-mix.

In the second mutation assay, the test item was tested up to concentrations of 512 and

5000 μg/plate in the absence and presence of 10% (v/v) S9-mix in the Salmonella

typhimurium and Escherichia coli strain, respectively. The test item precipitated on the plates

at the dose level of 5000 μg/plate. Cytotoxicity, as evidenced by a decrease in the number of

revertants, reduction of the bacterial background lawn and or the presence of microcolonies,

was observed in all tester strains in the absence and presence of S9-mix. Except for tester

strain WP2uvrA in the presence of S9-mix where no toxicity was observed.

BOURGEONAL did not induce a significant dose-related increase in the number of revertant

(His+) colonies in each of the four tester strains (TA1535, TA1537, TA98 and TA100) and in

the number of revertant (Trp+) colonies in tester strain WP2uvrA both in the absence and

presence of S9-metabolic activation. These results were confirmed in an independently

repeated experiment.

In this study, acceptable responses were obtained for the negative and strain-specific positive

control items indicating that the test conditions were adequate and that the metabolic

activation system functioned properly.

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

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

reverse mutation assay.