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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:
June - September 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 guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
GLP compliance:
yes (incl. certificate)
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid: viscous
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material:
16JSVA015
- Expiration date of the lot/batch:
14. September 2018
- Purity test date:
not state

RADIOLABELLING INFORMATION (if applicable)
not applicable

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material:
Room Temperature: (20 ± 5°C), keep container tightly closed, store under inert gas
- Stability under test conditions:
assumed stable


TREATMENT OF TEST MATERIAL PRIOR TO TESTING
none

Method

Target gene:
his-
Species / strain
Species / strain / cell type:
S. typhimurium, other: TA97a, TA98, TA100, TA102, TA1535
Metabolic activation:
with and without
Metabolic activation system:
rat liver S9
Test concentrations with justification for top dose:
The stock solution was used to prepare the geometric series of the concentrations to be
tested. The following nominal concentrations were prepared for the first experiment:
5 μL/plate, 1.5 μL/plate, 0.5 μL/plate, 0.15 μL/plate and 0.05 μL/plate.
The following nominal concentrations were prepared for the second experiment:
5 μL/plate, 2.5 μL plate, 1.25 μL/plate, 0.63 μL/plate, 0.31 μL/plate, 0.16 μL/plate and
0.08 μL/plate.
Vehicle / solvent:
In a non-GLP pre-test, the solubility of the test item was determined in a concentration of
50 mL/L in demineralized water, dimethyl sulfoxide (DMSO) and ethanol.
The test item was soluble in a concentration of 50 mL/L in ethanol.
Based on these results, ethanol was used as solvent in the experiments.
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
sodium azide
benzo(a)pyrene
other: 4-Nitro-1,3-phenylene Diamine, 2-Amino-Anthracene
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation); preincubation;
- Cell density at seeding (if applicable): not applicable

DURATION
- Preincubation period: 20 min
- Exposure duration: 48 h
- Expression time (cells in growth medium): not applicable
- Selection time (if incubation with a selection agent): not applicable
- Fixation time (start of exposure up to fixation or harvest of cells): not applicable

SELECTION AGENT (mutation assays): not applicable

SPINDLE INHIBITOR (cytogenetic assays): not applicable

STAIN (for cytogenetic assays): not applicable

NUMBER OF REPLICATIONS: 3

METHODS OF SLIDE PREPARATION AND STAINING TECHNIQUE USED: not applicable

NUMBER OF CELLS EVALUATED: not applicable

NUMBER OF METAPHASE SPREADS ANALYSED PER DOSE (if in vitro cytogenicity study in mammalian cells): not applicable

CRITERIA FOR MICRONUCLEUS IDENTIFICATION: not applicable

DETERMINATION OF CYTOTOXICITY
- Method: Number of colonies

OTHER EXAMINATIONS:
none

- OTHER:
Rationale for test conditions:
according to Guideline
Evaluation criteria:
A substance is considered to have mutagenic potential, if a reproducible increase of revertant
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:
none

Results and discussion

Test results
Key result
Species / strain:
S. typhimurium, other: TA97a, TA98, TA100, TA102, TA1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid

Applicant's summary and conclusion

Conclusions:
Based on the results of this study it is concluded that
Bis(neodecanoyloxy)dioctylstannane is not mutagenic in the Salmonella typhimurium
strains TA97a, TA98, TA100, TA102 and TA1535 in the absence and presence
of metabolic activation under the experimental conditions in this study.
Executive summary:

Two valid experiments were performed.

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

EC guidelines.

The test item Bis(neodecanoyloxy)dioctylstannane was tested in the Salmonella typhimurium

reverse mutation assay with five strains of Salmonella typhimurium (TA97a, TA98,

TA100, TA102 and TA1535).

The test was performed in two experiments in the presence and absence of metabolic

activation, with +S9 standing for presence of metabolic activation, and –S9 standing for

absence of metabolic activation.

In the first experiment, the test item (dissolved in ethanol) was tested up to concentrations

of 5 μL/plate in the absence and presence of S9-mix in the strains TA97a, TA98, TA100,

TA102 and TA1535 using the plate incorporation method.

The test item showed no precipitates on the plates at any of the concentrations.

The bacterial background lawn was not reduced at any of the concentrations and no relevant

decrease in the number of revertants was observed in all bacteria strains. The test item

showed no signs of toxicity towards the bacteria strains in both the absence and presence

of metabolic activation.

The results of this experiment showed that none of the tested concentrations showed a

significant increase in the number of revertants in all tested strains, in the presence and the

absence of metabolic activation.

Based on the first experiment, the test item was tested up to concentrations of 5 μL/plate in

the absence and presence of S9-mix in all bacteria strains using the pre-incubation method

(= second experiment).

The test item showed no precipitates on the plates at any of the concentrations.

No bacterial background lawn and no bacteria growth could be observed at the highest concentration

(5 μL/plate) in two bacteria strains (TA97a and TA102). The lower concentrations

were not affected.

The results of this experiments showed that the test item caused no increase in the number

of revertants in all bacteria strains compared to the solvent control, in both the absence and

presence of metabolic activation. The test item did not induce a dose-related increase in the

number of revertants colonies in all strains, in the presence and absence of metabolic activation.