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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:
19 Jan 2017 - 27 Mar 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 guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
1997
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Reference substance name:
Butyl 2-methylcrotonate
EC Number:
232-084-1
EC Name:
Butyl 2-methylcrotonate
Cas Number:
7785-64-0
Molecular formula:
C9H16O2
IUPAC Name:
butyl (2E)-2-methylbut-2-enoate
Constituent 2
Reference substance name:
3-methylbutyl 2-methylbut-2-enoate
Cas Number:
66917-62-2
Molecular formula:
C10H18O2
IUPAC Name:
3-methylbutyl 2-methylbut-2-enoate
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source of test material: Berjé Incorporated, 700 Blair Road, Carteret NJ 07008, USA.
- Lot/batch No.of test material: R-92286
- Expiration date of the lot/batch: 14 September 2018.
- Purity test date: 14 September 2016.

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature (cool, away from heat and sources of ignition)

Method

Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Remarks:
E. coli WP2uvrA (pKM101)
Metabolic activation:
with and without
Metabolic activation system:
Aroclor 1254/rat liver-derived (S9)
Test concentrations with justification for top dose:
Test 1: 0, 1.6, 5, 16, 50, 160, 500, 1600 and 5000 ug/plate
Test 2: 0, 5, 16, 50, 160, 500, 1600 and 5000 ug/plate.
Highest tested concentration is guideline limit dose.
Vehicle / solvent:
DMSO
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
9-aminoacridine
2-nitrofluorene
sodium azide
other: 2-Aminoanthracene; potassium dichromate
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Incubation period: 3 days

NUMBER OF REPLICATIONS: All tests (solvent controls, test item and positive controls) were carried out in triplicate

- OTHER: The number of revertant colonies was scored using a Sorcerer Colony Counter, except in cases where the mean incidence of revertant colonies on the solvent control plates was 20 or less (typically with strains TA1535 and TA1537).
Evaluation criteria:
A test item was considered to be mutagenic if the following criteria were satisfied:

1. For TA1535 or TA1537, the mean number of revertant colonies of one or more doses of the test item, with or without metabolic activation was equal to or greater than 2 times the concurrent solvent control mean value and the relevant historical mean value; for any other strain, the mean number of revertant colonies was equal to or greater than 2 times the concurrent solvent control mean value in the presence of one or more doses of the test item, with or without metabolic activation.

2. There was a dose-related increase in the number of revertant colonies.

3. Any increase was reproducible.

Results and discussion

Test resultsopen allclose all
Key result
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Key result
Species / strain:
E. coli WP2 uvr A pKM 101
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Remarks on result:
other: Mutagenicity Test 1

Applicant's summary and conclusion

Conclusions:
Anthemis nobilis, ext., (Chamomile oil) was not mutagenic in an OECD guideline- and GLP-compliant bacterial reverse mutation assay using Salmonella typhimurium strains TA98, TA100, TA1535 and TA1537 and Escherichia coli strain WP2uvrA (pKM101) when tested at up to cytotoxic and precipitating concentrations, with and without metabolic activation.
Executive summary:

Anthemis nobilis, ext., (Chamomile oil) was tested in a guideline (OECD 471)- and GLP-compliant bacterial reverse mutation assay. The substance was tested at up to 5 mg/plate using a plate incorporation method, with Salmonella typhimurium strains TA98, TA100, TA1535 and TA1537 and Escherichia coli strain WP2uvrA (pKM101), with and without a rat liver -derived metabolic activation fraction. The test material was not mutagenic under the conditions of this assay.