Fatty acids, C12-18 and C18-unsatd., reaction products with N,N-dimethyl-1,3-propanediamine and triethanolamine

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

28.02.2019 - 15.04.2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

Adopted July 21, 1997

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids, C12-18 and C18-unsatd., reaction products with N,N-dimethyl-1,3-propanediamine and triethanolamine

Cas Number:

91845-03-3

Molecular formula:

Unknown

IUPAC Name:

Fatty acids, C12-18 and C18-unsatd., reaction products with N,N-dimethyl-1,3-propanediamine and triethanolamine

Test material form:

solid: bulk

Specific details on test material used for the study:

- Source and package No.of test material: lot #19953859
- Expiration date of the lot/batch: 16.01.2020

Method

Target gene:

Mutagenicity test with Salmonella typhimurium according to OECD 471 (21 July 1997) with five tester strains (TA 98, TA 100, TA 102, TA 1535 and TA 1537) with and without metabolic activation of Aroclor induced rat liver S9.

Species / strain

Species / strain / cell type:

other: TA98, TA100, TA102, TA1535, TA1537

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

Aroclor rat liver S9

Test concentrations with justification for top dose:

Without metabolic activation: 0.1, 0.15, 1.5, 5, 45 µg/plate
Without metabolic activation: 1.5, 2, 2.5 µg/plate

Vehicle / solvent:

DMSO

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

yes

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

benzo(a)pyrene

cumene hydroperoxide

other: 2-aminoanthracene (2AA); and ICR-191 (Sigma)

Details on test system and experimental conditions:

In the present study the strains TA98, TA100, TA102, TA1535 and TA1537 were used. The strains TA98 and TA100 were supplied by German Federal Environment Office (UBA, Dessau).The strains TA102, TA1535 and TA1537 were supplied by Moltox Inc., Boone, USA imported by Trinova Biochem GmbH, D-35394 Gießen. The Aroclor induced rat liver S9 extract was also obtained from Moltox Inc., Boone, USA imported by Trinova Biochem GmbH, D-35394 Gießen. All Salmonella strains applied in the Ames test are derived from S. typhimurium type LT2.
Every strain is characterized by a mutation in the histidine operon. Additionally, the different
strains all bear the following two mutations which increase their susceptibility against
mutagenic agents considerably:

Rationale for test conditions:

According to OECD guideline

Evaluation criteria:

The colonies were counted manually with a colony counter (Heathro Scientific LLC). The mean value and standard deviation of the three resp. five replicates were calculated (Microsoft Excel ® ). The resulting induction rates (IR = number of revertant colonies in the plate with test substance / mean number of revertant colonies in the negative control plate [NC]) were calculated for each test concentration and each tester strain. Cytotoxic effects of the test item can be determined if the growth of the micro colonies in the background lawn deviates from the negative control plates. A decrease in micro colonies density or gaps in the bacterial background lawn are indicators for cytotoxic effects. These are discovered by microscopic evaluation of the plates. Often such effects can be observed in combination with reduced number of revertant colonies in comparison to the number of spontaneous revertant colonies on the negative control plates (induction rate < 1). Toxic effects to the bacteria are reported. For the evaluation of the results of the tests performed in this study the increase of revertant colonies in a sample is expressed as the induction rate [IR], (IR = number of revertant
colonies in the sample/number of revertant colonies in the control). In the case of a reproducible increase of the number of revertant colonies and if additionally a statistically significant difference between the mean values can be demonstrated, for example with the U-test according to Mann & Whitney (Wahrendorf et al. 1985), the sample is evaluated as positive.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 1535

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

Test results 2

Key result

Species / strain:

S. typhimurium TA 1537

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

Test results 3

Key result

Species / strain:

S. typhimurium TA 98

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

Test results 4

Key result

Species / strain:

S. typhimurium TA 100

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

Test results 5

Key result

Species / strain:

S. typhimurium TA 102

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

Additional information on results:

The colonies were counted manually with a colony counter (Heathro Scientific LLC). The mean value and standard deviation of the three resp. five replicates were calculated (Microsoft Excel ® ). The resulting induction rates (IR = number of revertant colonies in the plate with test substance / mean number of revertant colonies in the negative control plate [NC]) were calculated for each test concentration and each tester strain.

Any other information on results incl. tables

Due  to  clear  cytotoxic  effects  with  the  highest  test  concentration  of  4.0  mg/plate  an independent repetition of the study with adapted test concentrations between 0.6 mg/plate and 3.0 mg/plate were applied. The results of the first and the second test (independent repetition) did not deviate. 

Applicant's summary and conclusion

Conclusions:

The test item 35760/270 is not mutagenic in the Ames Test according to OECD 471.

Executive summary:

In none of the tester strains a dose dependent increase of revertant colonies and no reproducible increase at one or more concentrations in the number of revertant colonies per plate with or without the metabolic activation occurred. The bacterial background lawn was affected up to 2.0 mg/plate. The results of the first and the second test (independent repetition) did not deviate. The test item 35760/270 is not mutagenic in the Ames Test according to OECD 471.