Fatty acids, C16-18 and C18-unsatd., branched and linear, reaction products with diethylenetriamine and tall-oil fatty acids, Me maleates

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

For the test substance the Ames test with Salmonella typhimurium strains TA 98, TA 100, TA 1535 and TA1537, as well as E-coli is negative with and without metabolic activation (Harlan 2012k). The substance did not induce mutations in the Mouse lymphoma assay (Harlan 2013i). No chromosome aberrations were found in a chromosome aberration test(Harlan 2013h) with and without metabolic activation

Link to relevant study records

Reference

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:

29-02-2012 to 09-06-2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study according to the guidelines, under GLP

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Qualifier:

according to guideline

Guideline:

OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Target gene:

Salmonella typhimurium: histidine
E-coli: tryptophan

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 from rats induced with Phenobarbitone/ß-Naphthoflavone

Test concentrations with justification for top dose:

50, 150, 500, 1500 and 5000 ug/plate

Vehicle / solvent:

acetone (100 mg/ml)

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

acetone

Positive controls:

yes

Remarks:

for TA100, TA1535 and WP2uvrA without metabolic activation

Positive control substance:

N-ethyl-N-nitro-N-nitrosoguanidine

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

acetone

Positive controls:

yes

Remarks:

for TA100, TA1535, TA 1537 and WP2uvrA with metabolic activation; for TA1537 without metabolic activation

Positive control substance:

2-acetylaminofluorene

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

dimethylformamide

Positive controls:

yes

Remarks:

for TA98 without metabolic acivation

Positive control substance:

4-nitroquinoline-N-oxide

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

acetone

Positive controls:

yes

Remarks:

for TA98 with metabolic activation

Positive control substance:

benzo(a)pyrene

Details on test system and experimental conditions:

METHOD OF APPLICATION: 1st test in agar (plate incorporation); 2nd test preincubation

DURATION
- Preincubation period: 20 min at 37°C
- Exposure duration: 48 hours at 37°C

NUMBER OF REPLICATIONS: 3/concentration

DETERMINATION OF CYTOTOXICITY
- Method: growth rate bacterial bachground lawn

COLONY COUNT:
Domino colony counter

Evaluation criteria:

A test item will be considered mutagenic (positive) in the test system if one or more of these criteria are met
1. A dose-related increase in mutant frequency over the dose range tested (De Serres and Shelby (1979)).
2. A reproducible increase at one or more concentrations.
3. Biological relevance against in-house historical control ranges.
4. Statistical analysis of data as determined by UKEMS (Mahon et al (1989)).
5. Fold increase greater than two times the concurrent solvent control for any tester strain (especially if accompanied by an out-of-historical range response).

Statistics:

Not specified according to UKEMS

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:

no cytotoxicity, but tested up to precipitating concentrations

Remarks:

precipitate at 5000 ug/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Remarks:

precipitate at 5000 ug/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

In the second test a slight but significant increase (p≤0.05) of the number of mutants was seen in E. coli WP2 uvr A with metabolic activation at 5000 ug/plate.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Conclusions:

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

The test substance did not induce mutations (base-pair substitution and frame-shift type) in Salmonella typhimurium strains TA98, TA100, TA1535 and TA1537 as well as in E. coli WP2 uvr A both in presence and absence of metabolic activation.

Executive summary:

The test substance was tested in an Ames test at concentrations upto the preciptation level (5000 ug/plate). In a plate incorporation and a pre-incubation assay (both performed in triplicate), the test substance did not induce mutations (base-pair substitution and frame-shift type) in Salmonella typhimurium strains TA98, TA100, TA1535 and TA1537, as well as in E. coli WP2 uvr A both in presence and absence of metabolic activation.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

Additional information from genetic toxicity in vitro:

The test substance was tested in an Ames test at concentrations upto the preciptation level (5000 ug/plate). In a plate incorporation and a pre-incubation assay (both performed in triplicate), the test substance did not induce mutations (base-pair substitution and frame-shift type) in Salmonella typhimurium strains TA98, TA100, TA1535 and TA1537, as well as in E. coli WP2 uvr A both in presence and absence of metabolic activation.

(Harlan 2012k).

The substance is considered to be non mutagenic in the mouse lymphoma thymidine kinase locus using the cell line L5178Y (Harlan 2013i) and no chromosomal changes were observed in a chromosome aberration test in Chinese hamster lung fibroblasts (V79) (Harlan2013h). Both tests were performed in presence and absence of metabolic activation.

Based on the negative outcome of the three tests, it is concluded that it is not expected that the test substance will induce mutagenic or clastogenic effects. No additional testing is considered necessary.

Justification for classification or non-classification

The test substance does not need to be classified for mutagenicity.