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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:
2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
guideline study under 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)
Qualifier:
according to guideline
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Reference substance name:
Addition product of maleic anhydride, tall oil fatty acids, linseed oil and methanol
IUPAC Name:
Addition product of maleic anhydride, tall oil fatty acids, linseed oil and methanol
Test material form:
liquid
Details on test material:
UVCB, Purity 100%, brown liquid, homogenous, Batch number 210150084 Expiry date 18 May 2016

Method

Target gene:
histidine
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
S9 rat liver metabolic system
Test concentrations with justification for top dose:
5000, 1580, 500, 158 and 50.0 μg/plate
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
not specified
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
9-aminoacridine
2-nitrofluorene
sodium azide
methylmethanesulfonate
other:
Details on test system and experimental conditions:
Solutions of the test item, as received, were prepared immediately before use in DMSO. Solutions were prepared on a weight/volume basis without correction for the displacement due to the volume of the test item, i.e. the test item was weighed and the necessary volume of solvent was added in order to reach the required concentration.

An initial assay using the plate incorporation method was utilised. The assay was repeated using the pre-incubation method.
Rationale for test conditions:
Solubility of the test item was evaluated in a preliminary trial using DMSO. This solvent was selected since it is compatible with the survival of the
bacteria and the S9 metabolic activity. The test item was found to be soluble at 100 mg/mL.This result permitted a maximum concentration of 5000 μg/plate to be used in the toxicity test.
Evaluation criteria:
Criteria for positive: A two-fold (or more) increase in mean revertant numbers must be observed at two consecutive doses or at the highest practicable dose only. Also, there must be evidence of a dose-response relationship.
Statistics:
Mean plus standard deviation, according to accepted methods.

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:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
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:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
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:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
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:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Key result
Species / strain:
E. coli WP2 uvr A
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:
not applicable
Positive controls validity:
valid

Any other information on results incl. tables

A slight decrease in the number of revertant colonies was observed at the
two highest dose levels tested for TA100 tester strain, in the absence and

presence of S9 metabolism. Otherwise, it did not cause reverse mutation.

This does not impact the conclusion that the substance is not mutagenic this assay.

Applicant's summary and conclusion

Conclusions:
It is concluded that the test item does not induce reverse mutation in Salmonella typhimurium TA 1535, TA 1537, TA 98 or TA 100 or Escherichia coli WP2 uvrA. in the absence or presence of S9 metabolism, under the reported standard experimental conditions of this OECD 471 assay.