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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:
30. Jun. 2015- 10. Jul. 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study with detailed documentation, in accordance with GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report date:
2015

Materials and methods

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

Test material

Constituent 1
Reference substance name:
MAEEBP
IUPAC Name:
MAEEBP
Details on test material:
- Name of test material (as cited in study report): MAEEBP- Physical state: Colourless liquid- Composition of test material, percentage of components: 4-methacryloyl diethoxy benzophenone (purity 85 area %)- Impurities: 4-hydroxybenzophenone <5%, 4-methacryloyl benzophenone <5%, polymethacrylate <5%- Lot/batch No.: 150301- Expiration date of the lot/batch: 01.09.2015- Storage condition of test material: Fridge: 2-8° C, keep away from light and humidity

Method

Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 98
Species / strain / cell type:
S. typhimurium TA 1535
Species / strain / cell type:
S. typhimurium TA 100
Species / strain / cell type:
S. typhimurium TA 102
Species / strain / cell type:
S. typhimurium, other: TA 97 a
Metabolic activation:
with and without
Metabolic activation system:
S9
Test concentrations with justification for top dose:
experiment 1: 50 µg/plate, 15 µg/plate, 5 µg/plate, 1.5 µg/plate and 0.5 µg/plate.experiment 2: 50 µg/plate, 25 µg/plate, 12.5 µg/plate, 6.3 µg/plate, 3.1 µg/plate and 1.6 µg/plate
Vehicle / solvent:
- vehicle for the test item: dimethyl sulfoxide (DMSO)- vehicle for the positive controls: DMSO (4-Nitro-1,2-phenylene Diamine, 2-Amino-anthracene, Benzo-a-pyrene) or demineralised water (Sodium azide)- vehicle for solvent controls: DMSO or demineralised water
Controlsopen allclose all
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Remarks:
strains TA97a, TA98, TA100, TA102, TA1535 with and without S9
Positive controls:
yes
Positive control substance:
other: 4-Nitro-1,2-phenylene diamine
Remarks:
strains TA97a, TA98 and TA102, no metabolic activation
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Remarks:
strains TA97a, TA98, TA100, TA102, TA1535 with and without S9
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
strains TA100 and TA1535, no metabolic activation
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Remarks:
strains TA97a, TA98, TA100, TA102, TA1535 with and without S9
Positive controls:
yes
Positive control substance:
other: 2-Amino-anthracene
Remarks:
strains TA97a, TA100, TA102 and TA1535, with metabolic activation
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Remarks:
strains TA97a, TA98, TA100, TA102, TA1535 with and without S9
Positive controls:
yes
Positive control substance:
benzo(a)pyrene
Remarks:
strain TA98, with metabolic activation
Details on test system and experimental conditions:
METHOD OF APPLICATION: plate incorporation in experiment 1, pre-incubation method in experiment 2- Experiment 1:Suspensions of bacterial cells are exposed to the test substance in the presence and in the absence of an exogenous metabolic activation system. In the plate incorporation method, these suspensions are mixed with an overlay agar and plated immediately onto minimal medium. In the pre-incubation method, the treatment mixture is incubated and then mixed with an overlay agar before plating onto minimal medium. After 2 or 3 days of incubation, revertant colonies are counted and compared to the number of spontaneous revertant colonies on solvent control plates.5 concentrations of the test item (0.5, 1.5, 5, 15, 50 µg/plate) dissolved in DMSO were used. 5 genetically changed strains of Salmonella typhimurium (TA97a, TA98, TA100, TA102 (genetically manipulated) and TA1535) were exposed to the test item both in the presence and in the absence of a metabolic activation system (S9-mix, rat liver S9-mix induced by Aroclor 1254) for 48 h, using the plate incorporation method.- Experiment 2: Suspensions of bacterial cells are exposed to the test substance in the presence and in the absence of an exogenous metabolic activation system. In the pre-incubation method, the treatment mixture is incubated and then mixed with an overlay agar before plating onto minimal medium. After 2 or 3 days of incubation, revertant colonies are counted and compared to the number of spontaneous revertant colonies on solvent control plates.6 concentrations of the test item (1.6, 3.1, 6.3, 12.5, 25, 50 µg/plate) dissolved in DMSO were used. 5 genetically changed strains of Salmonella typhimurium (TA97a, TA98, TA100, TA102 (genetically manipulated) and TA1535) were exposed to the test item both in the presence and in the absence of a metabolic activation system (S9-mix, rat liver S9-mix induced by Aroclor 1254) for 48 h, using the pre-incubation method.
Evaluation criteria:
The mean values and standard deviations of each threefold determination of revertant colonies are calculated as well as the increase factor of revertant induction (mean revertants divided by mean spontaneous revertants) of the test item solutions and the positive controls. A substance is considered to have mutagenic potential, if a reproducible increase of re-vertant colonies per plate exceeding an increase factor of 2 in at least one strain can be observed. A concentration-related increase over the range tested is also taken as a sign of mutagenic activity.
Statistics:
The mean values and standard deviations of each threefold determination of revertant colonies are calculated as well as the increase factor of revertant induction (mean revertants divided by mean spontaneous revertants) of the test item solutions and the positive controls.

Results and discussion

Test results
Species / strain:
other: TA97a, TA98, TA100, TA102, TA1535
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

Any other information on results incl. tables

- Experiment 1:

None of the concentrations caused a significant increase in the number of revertant colonies in the tested strains. The test item did not show any mutagenic effects in the first experiment.

The test item showed no precipitates on the plates in all tested concentrations

No signs of toxicity towards the bacteria could be observed.

The sterility control and the determination of the titre did not show any inconsistencies. The determined values for the spontaneous revertants of the negative controls were in the normal range, with exception of strain TA97a with and without S9. Both values lay outside the range of historical laboratory control data. The deviations of negative controls were not confirmed in the respective independent second experiment. All positive controls showed mutagenic effects with and without metabolic activation. - Experiment 2: The test item did not show mutagenic effects in the second experiment, either.

The test item showed no precipitates on the plates in all tested concentrations.

No signs of toxicity towards the bacteria could be observed.

The sterility control and the determination of the titre did not show any inconsistencies. The determined values for the spontaneous revertants of the negative controls were in the normal range. All positive controls showed mutagenic effects with and without metabolic activation. However, the experimental value of the positive control using strain TA1535 without S9 deviated from historical data of the laboratory. This deviation was not confirmed in the results of the first experiment.

 

Under the conditions of experiment 1 and experiment 2, the test item did not show mutagenic effects towardsSalmonella typhimurium,strainsTA97a, TA98, TA100, TA102 and TA1535.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):negativeUnder the conditions of the test, the test item did not show mutagenic effects towards Salmonella typhimurium, strains TA97a, TA98, TA100, TA102 and TA1535. Therefore, no concentration-effect relationship could be determined.The test item MAEEBP is considered as “not mutagenic under the conditions of the test”.

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