Registration Dossier

Diss Factsheets

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
not specified
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
To address toxicological endpoints as part of the REACH registration of the REACH registration of Amyl Salicylate (Target Substance) it is proposed to read-across to Benzyl Salicylate (Source Substance). The use of read-across works within the spirit of REACH and the stated aim of the legislation to reduce animal testing where possible. The Target Substance and Source Substance have been characterised using the categories and databases present in the OECD [Q]SAR Toolbox. From the profiling, it can be seen that the two substances share structural similarities and also ‘mechanistic action’ similarities which are both general and endpoint specific. Therefore read across is justified.
Justification for type of information:
The justification for read across from Benzyl Salicylate is attached in section 13.2 (Other assesment report).

Data source

Reference
Reference Type:
publication
Title:
Unnamed
Year:
1987
Report date:
1987

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
other: Haworth S, Lawlor T, Mortelmans K, Speck W, Zeiger E (1983): Salmonella mutagenicity test results for 250 chemicals. Environ Mutagen S [Suppl 1]:3-142
Principles of method if other than guideline:
Ames test.
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
Benzyl salicylate
EC Number:
204-262-9
EC Name:
Benzyl salicylate
Cas Number:
118-58-1
Molecular formula:
C14H12O3
IUPAC Name:
benzyl salicylate
Details on test material:
- Name of test material (as cited in study report): Benzyl salicylate
- Substance type: No data
- Physical state: No data
- Analytical purity: No data
- Impurities (identity and concentrations): No data
- Composition of test material, percentage of components: No data
- Isomers composition: No data
- Purity test date: No data
- Lot/batch No: No data
- Expiration date of the lot/batch: No data
- Stability under test conditions: No data
- Storage condition of test material: No data

Method

Target gene:
Histidine
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Metabolic activation system:
S9 mixes from rat (RLI) and hamster (HLI), tested separately.
Test concentrations with justification for top dose:
EGG: 0, 0.3, 1, 3.3, 10, 20, 33, 100, 333 and 666 µg/plate.
SRI: 0, 3.3, 10, 33.3, 100 and 333 µg/plate.
Vehicle / solvent:
- Vehicle/solvent used: DMSO
- Justification for choice of solvent/vehicle: Assessed by each individual laboratory from a choice of distilled water, DMSO, 95 % ethanol, acetone, in that order of preference.
Controlsopen allclose all
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
Positive controls:
yes
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
TA1535 -S9 and TA100 -S9 only
Positive control substance:
9-aminoacridine
Remarks:
TA1537 -S9 only
Positive control substance:
other: 4-nitro-o-phenylenediamine
Remarks:
TA98 -S9 only
Positive control substance:
other: 2-aminoanthracene
Remarks:
TA 1535 +S9, TA 1537 +S9, TA 98 +S9 and TA 100 +S9 only
Details on test system and experimental conditions:
The test chemical, Salmonella culture, and S-9 mix or buffer were incubated
at 37"C, without shaking, for 20 min. Chemicals known or suspected to be volatile
were incubated in capped tubes. The top agar was added, and the contents of the
tubes were mixed and poured onto the surface of petri dishes that contained Vogel-
Bonner medium [Vogel and Bonner, 19561. The histidine-revertant (his') colonies
arising on these plates were counted following 2 days incubation at 37°C. The plates

METHOD OF APPLICATION: in medium; in agar (plate incorporation); preincubation; in suspension; as impregnation on paper disk

DURATION
- Preincubation period:
- Exposure duration:
- Expression time (cells in growth medium):
- Selection time (if incubation with a selection agent):
- Fixation time (start of exposure up to fixation or harvest of cells):

NUMBER OF REPLICATIONS:

NUMBER OF CELLS EVALUATED:

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other:

OTHER EXAMINATIONS:
- Determination of polyploidy:
- Determination of endoreplication:
- Other:

OTHER:
Evaluation criteria:
As per Haworth et al. 1983:

An individual trial was judged mutagenic (+) if a dose-related increase over the corresponding solvent control was seen, and it was judged weakly mutagenic C+W) if a low-level dose response was seen. A trial was considered questionable (?) if a dose-related increase was judged insufficiently high to justify a call of " + W," if only a single dose was elevated over the control, or if a non-dose-related increase was seen.

The distinctions between a weak mutagenic response and a mutagenic response, or between a weak mutagenic response and a questionable mutagenic response were considered highly subjective.

A chemical was judged to be mutagenic (+), or weakly mutagenic (+W), if it produced a reproducible, dose-related increase in his⁺ revertants over the corresponding solvent controls in replicate trials. A chemical was considered to be questionable (?) if a reproducible increase of hist revertants did not meet the criteria for either a " + " or " + W," or if only single doses produced an increase in his⁺ revertants in repeat trials. The chemicals were decoded by the chemical repository only after a determination had been made regarding their mutagenicity or nonmutagenicity.

Results and discussion

Test results
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:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
For full results please refer to the attached supporting information.
Remarks on result:
other: all strains/cell types tested

Any other information on results incl. tables

To address toxicological endpoints as part of the REACH registration of the REACH registration of Amyl Salicylate (Target Substance) it is proposed to read-across to BenzylSalicylate (Source Substance).

 

The use of read-across works within the spirit of REACH and the stated aim of the legislation to reduce animal testing where possible.

 

The Target Substance and Source Substance have been characterised using the categories and databases present in the OECD [Q]SAR Toolbox. From the profiling, it can be seen that the two substances share structural similarities and also ‘mechanistic action’ similarities which are both general and endpoint specific.

See Section 13, document Read across justification_Tox_Amyl salicylate_Benzyl salicylate.

Applicant's summary and conclusion

Conclusions:
Zeiger et al. screened 255 chemicals using Ames tests, including benzyl salicylate, which was screened in two separate tests in two different laboratories. Both test found benzyl salicylate to be negative for genotoxicity.
Data are taken from results of study on the structural analgue, Benzyl salicylate, EC# 204-262-9. Read across is justified based on similarity in chemical structure and biological activity of salicylates.