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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:
November 06, 2013 - March 03, 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report Date:
2014

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Qualifier:
according to
Guideline:
EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
GLP compliance:
yes (incl. certificate)
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Specific details on test material used for the study:
Name of test material: Art. 201386
Synonym: B-2O-O5
Chemical name: 3-Ethoxy-4,6-difluoro-7-pentyloxy-dibenzofuran
Batch No.: EF13003186
Purity (GC): 99.02 %
Appearance: white solid
Supplier: Merck KGaA, Darmstadt
Released until: October 31, 2014

Method

Target gene:
HIS Operon, TRP Operon
Species / strain
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Metabolic activation:
with and without
Metabolic activation system:
liver S9 mix obtained from rats pretreated with Aroclor 1254
Test concentrations with justification for top dose:
1st series: 5.00, 15.8, 50.0, 158, 500, 1580, 5000 µg/plate
2 nd series: 5.00, 15.8, 50.0, 88.9, 158, 281 µg/plate
Vehicle:
Acetone
Controls
Negative controls:
yes
Solvent controls:
yes
Positive controls:
yes
Positive control substance:
other: Without S9: Sodium azide 9-Aminoacridine Daunomycin 4-Nitroquinoline-N-oxide with S9: 2-Aminoanthracene
Details on test system and conditions:
METHOD OF APPLICATION: in medium; in agar plate incorporation
- Cell density at seeding (if applicable): approx. 1 e9

DURATION
- Exposure duration: 2-3 days

SELECTION AGENT (mutation assays): histidine

STAIN (for cytogenetic assays):

NUMBER OF REPLICATIONS: neg. controls: 6, test item: 3, pos control: 3

DETERMINATION OF CYTOTOXICITY
reduction of background lawn

OTHER EXAMINATIONS:
Precipitation

- OTHER:
Rationale for test conditions:
According to guideline
Evaluation criteria:
According to guideline
Statistics:
Standard statistical methods have been applied for data processing.

Results and discussion

Test results
Key result
Species / strain:
other: all strains / all conditions
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
no, but tested up to precipitating concentrations
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Additional information on results:
Precipitation of the test material on the agar plates occurred at concentrations ≥ 158 µg/plate.

Any other information on results incl. tables

Objective

The present study was conducted to investigate the test material for mutagenic potential in a bac-terial reverse gene mutation assay.

Study Design

The investigations for the mutagenic potential of Art. 201386 were performed using Salmonella typhimurium tester strains TA 98, TA 100, TA 1535 and TA 1537, and Escherichia coli WP2 uvrA. The plate incorporation test with and without addition of liver S9 mix from Aroclor 1254-pretreated rats was used. Two independent experimental series were performed. In the two series with S9 mix, 10 % S9 in the S9 mix were used in the 1st and 30 % in the 2nd series, respectively.

This study was performed according to GLP and the methods applied are fully compliant with OECD TG 471.

Results

Art. 201386 was dissolved in acetone and tested at concentrations ranging from 5.00 to 5000 µg/plate. Precipitation of the test material on the agar plates occurred at concentrations ≥ 158 µg/plate. Toxicity to the bacteria was not observed.

Daunomycin, sodium azide, 9-aminoacridine and 4-nitroquinolin-N-oxide served as strain specif-ic positive control test materials in the absence of S9 mix. 2-Aminoanthracene was used for test-ing the bacteria and the activity of the S9 mix. Each treatment with the test materials used as positive controls led to a clear increase in revertant colonies, thus, showing the expected reversion properties of all strains and good metabolic activity of the S9 mix used.

Following Art. 201386 treatments of all the tester strains in the absence and presence of S9 mix, no relevant increases in revertant numbers were observed.

Conclusion

With and without addition of S9 mix as the external metabolizing system, Art. 201386 was not mutagenic under the experimental conditions described.

Applicant's summary and conclusion

Conclusions:
With and without addition of S9 mix as the external metabolizing system, Art. 201386 was not mutagenic under the experimental conditions described.
Executive summary:

With and without addition of S9 mix as the external metabolizing system, Art. 201386 was not mutagenic under the experimental conditions described.