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Toxicological information

Genetic toxicity: in vitro

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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:
15 April 2010 - 26 april 2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Deviations:
no
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Ebecryl® P116 radiation curing resins- Substance type: Clear colourless liquid- Physical state: Liquid- Stability under test conditions: Stable- Storage condition of test material: At room temperature protected from light

Method

Target gene:
S. typhimurium: Histidine geneE. coli: Tryptophan gene
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
Rat liver S9-mix induced by a combination of phenobarbital and β-naphthoflavone
Test concentrations with justification for top dose:
Experiment 1:Preliminary test (with and without S9) TA100 and WP2uvrA: 3; 10; 33; 100; 333; 1,000; 3,330 and 5,000 µg/plate.Main study: TA1535, TA1537 and TA98Without and with S9-mix: 100; 333; 1,000; 3,330 and 5,000 µg/plate.Experiment 2:Without and with S9-mix: 100; 333; 1,000; 3,330 and 5,000 µg/plate.
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO- Justification for choice of solvent/vehicle: The test substance was soluble in DMSO. DMSO is accepted and approved by authorities and international guidelines.
Controlsopen allclose all
Negative solvent / vehicle controls:
yes
Remarks:
(DMSO)
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
without S9 Migrated to IUCLID6: (5 µg/plate in saline for TA 1535)
Positive control substance:
9-aminoacridine
Remarks:
without S9 Migrated to IUCLID6: (60 µg/plate in water for TA1537)
Positive control substance:
2-nitrofluorene
Remarks:
without S9 Migrated to IUCLID6: (10 µg/platein DMSO for TA98)
Positive control substance:
methylmethanesulfonate
Remarks:
without S9 Migrated to IUCLID6: (6650 µg/platein DMSO TA100)
Positive control substance:
4-nitroquinoline-N-oxide
Remarks:
without S9 Migrated to IUCLID6: (10 µg/plate in DMSO for WP2uvrA)
Positive control substance:
other: 2-aminoanthracene inDMSO for all tester strains
Remarks:
with S9
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium; in agar (plate incorporation)DURATION:- Exposure duration: 48 hoursNUMBER OF REPLICATIONS:- Doses of the test substance were tested in triplicate in each strain. Two independent experiments were conducted.NUMBER OF CELLS EVALUATED: 10E8 per plateDETERMINATION OF CYTOTOXICITY- Method: Reduction of the bacterial background lawn, increase in the size of the microcolonies and reduction of the revertant colonies.OTHER EXAMINATIONS:- The presence of precipitation of the test substance on the plates was determined.
Evaluation criteria:
A test substance was considered negative (not mutagenic) in the test if:- The total number of revertants in tester strain TA100 was not greater than two (2) times the concurrent control, and the total number of revertants in tester strains TA 1535, TA1537, TA98 or WP2uvrA was not greater than three (3) times the concurrent control.- The negative response should have been reproducible in at least one independently repeated experiment.A test substance was considered positive if:- A two-fold (TA100) or more or a three-fold (TA1535, TA1537, TA98, WP2uvrA) or more increase above solvent control in the mean number of revertant colonies was observed in the test substance group.- When a positive response was observed in one of the tester strains, the positive response should have been reproducible in at least one independently repeated experiment.

Results and discussion

Test resultsopen allclose all
Key result
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
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS- Precipitation: No precipitation was observed up to and including the top dose of 5,000 µg/plate.RANGE-FINDING/SCREENING STUDIES:- No toxicity or mutagenicity was observed up to and including the top dose of 5,000 µg/plate.COMPARISON WITH HISTORICAL CONTROL DATA:- The negative and strain-specific positive control values were within our laboratory historical control data ranges indicating that the test conditions were adequate and that the metabolic activation system functioned properly.ADDITIONAL INFORMATION ON CYTOTOXICITY:- No toxicity or mutagenicity was observed up to and including the top dose of 5,000 µg/plate.

Applicant's summary and conclusion

Conclusions:
Under the test conditions, the test substance was found to be non-mutagenic in the bacterial reverse mutation assay.
Executive summary:

A study was conducted to determine the mutagenic potential of the test substance according to OECD Guideline 471 and EU Method B.13/14.

Tester strains TA98, TA100, TA1535, TA1537 of Salmonella typhimurium strain and WP2uvrA of Escherichia coli in the presence and absence of S9 were exposed to the test substance. The assay was performed in two phases, using the plate incorporation method. The first phase, the initial toxicity-mutation assay, was used to established the dose-range for the confirmatory mutagenicity assay and to provide a preliminary mutagenicity evaluation. The second phase, the confirmatory mutagenicity assay, was used to evaluated and confirm the mutagenic potential of the test substance.

DMSO was selected as the solvent based on its solubility of the test substance and compatibility with the target cells. In the initial toxicity-mutation assay, the dose levels tested were 3; 10; 33; 100; 333; 1,000; 3,330 and 5,000 µg/plate. In the main study, the dose levels tested were 100; 333; 1,000; 3,330 and 5,000 µg/plate.

The negative and 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.

Under the test conditions, the test substance was found to be non-mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay.