Registration Dossier

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:
11. Oct. 2012 -07.11.2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: well documented guideline study under GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report Date:
2012

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)
GLP compliance:
yes (incl. certificate)
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid
Details on test material:
Name: Initiator 94
Batch: 004IN11
according to certificate of analysis and its addendum of 1. August 2012:
purity: 32% (main component and active species: benzpinakolsilylethers
further composition: Phosphoric acid tributylester (Tributylphosphat) 19.3 %,
1,2-Benzenedicarboxylic acid-di-2-propenylester (Diallylphthalate) 8.7%;
MEthylbenzene (toluol) 12.9%; Diphenylmethanone (Benzophenon) 12.4 %;
1,1,1,1-Tetraphenylethane-1,2-diol (Benzpinakol) 14.6%
form: liquid
colour: brown
odour: aromatic
density: 1.13 g/cm3 at 20°C
Production date of batch: 16.05.2011
Expiry date of batch: 01.02.2013

Method

Species / strain
Species / strain / cell type:
other: TA 97a, TA 98, TA 100, TA 102, TA 1535
Metabolic activation:
with and without
Metabolic activation system:
S9
Test concentrations with justification for top dose:
First experiment: 0, 0.05, 0.151, 0.502, 1.505, 5.016 mg/plate,
second experiment: 0, 0.313, 0.626, 1.251, 2.501, 5.001 mg/plate
Vehicle / solvent:
DMSO, (Stock solution of 50 g/L of test item in DMSO)
DMSO was chosen as vehicle, because the test item was completely soluble and this solvent doesn't have any effects on the viability of the bacteria or the number of spontaneous revertants.
Controls
Untreated negative controls:
yes
Remarks:
DMSO and water
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
yes
Remarks:
water
Positive controls:
yes
Remarks:
4-Nitro-1,2-phenylene diamine, Sodium azide, 2-Amino-anthracene, Benz-a-pyrene
Details on test system and experimental conditions:
METHOD OF APPLICATION: in a fist test in agar (plate incorporation) and in a second test in suspension in the pre-incubation method;
DURATION
- Preincubation period: 20 minutes
- Exposure duration: 48 hours

NUMBER OF REPLICATIONS: 4

NUMBER OF CELLS EVALUATED:

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth;
Evaluation criteria:
Number of spontaneous revertants and positive control values in the range of the histrorical data.

Results and discussion

Test results
Species / strain:
other: TA 97a, TA 98, TA 100, TA 102, TA 1535 from TRINOVA BioChem
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
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative with and without metabolic activation

The test item Initiator 94 is considered as "not mutagenic under the conditions of the test".
Executive summary:

The mutagenic potential of Initiator 94 is determined with the Bacterial Reverse Mutation Test according to OECD 471/ EU-guideline B.13/14. Two valid experiments were performed, a first one using the plate incorporation method and a second one with the pre-incubation method. In both experiments five strains of Salmonella typhimurium: TA 97a, TA 98, TA 100, TA 102 and TA 1535, were used with and without metabolic activation by S9. Per strain and dose 4 plates with and 4 plates without metabolic activation were used.

In the first experiment five concentrations of the test item, ranging from 50 to 5015 µg/plate, were used and incubated for 48 hours.

In the second experiment five concentrations of the test item, ranging from 313 to 5001 µg/plate, were used and incubated for 48 hours.

In both experiments no signs of toxicity towards the bacteria and no mutagenic effects could be observed. The sterility control, the negative controls and the positive controls were within the range of historical control data.

Therefore, under the conditions of this test, the test item is considered not mutagenic.