N,N-bis(2-hydroxyethyl)dodecanamide

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:

Not available

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Cross-reference

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

1999

Materials and methods

Principles of method if other than guideline:

Suspensions of bacterial cells were exposed to the test substance by the plate incorporation method in the presence and in the absence of an exogenous metabolic activation system. The suspensions were mixed with an overlay agar and plated immediately onto the minimal medium and incubated for 2 d at 37˚C. The results were interpreted by counting the revertant colonies and comparing to the number of spontaneous revertant colonies on solvent-control plates.

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

impurity 1

Reference substance name:

Unknown impurities

IUPAC Name:

Unknown impurities

Constituent 1

Reference substance name:

N,N-bis(2-hydroxyethyl)dodecanamide

EC Number:

204-393-1

EC Name:

N,N-bis(2-hydroxyethyl)dodecanamide

Cas Number:

120-40-1

Molecular formula:

C16H33NO3

IUPAC Name:

N,N-bis(2-hydroxyethyl)dodecanamide

impurity 2

Reference substance name:

2,2'-iminodiethanol

EC Number:

203-868-0

EC Name:

2,2'-iminodiethanol

Cas Number:

111-42-2

Molecular formula:

C4H11NO2

IUPAC Name:

2,2’-iminodiethanol

Test material form:

other: Viscous light yellow liquid or a white to light yellow, waxy solid

Method

Species / strain

Species / strain / cell type:

other: S. typhimurium TA97, TA98, TA100 and TA1535

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

S9 mix (metabolic activation enzymes and cofactors from Aroclor 1254-induced male Sprague-Dawley rat or Syrian hamster liver).

Test concentrations with justification for top dose:

- without metabolic activation:0.0, 0.3, 1.0, 3.0, 10.0, 33.0, 66.0, 100.0, 333.0 µg/plate
- with metabolic activation: 0.0, 1.0, 3.0, 10.0, 33.0, 100.0, 166.0, 333.0, 1,000.0 µg/plate

Vehicle / solvent:

Ethanol

Controls

Controls 1

Untreated negative controls:

yes

Remarks:

solvent control

Negative solvent / vehicle controls:

yes

Remarks:

Ethanol

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

-S9 : for strains TA100 and TA1535

Controls 2

Untreated negative controls:

yes

Remarks:

solvent control

Negative solvent / vehicle controls:

yes

Remarks:

Ethanol

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

-S9 : for strain TA97

Controls 3

Untreated negative controls:

yes

Remarks:

solvent control

Negative solvent / vehicle controls:

yes

Remarks:

Ethanol

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: 4-nitro-o- phenylenediamine

Remarks:

-S9 : for strain TA98

Controls 4

Untreated negative controls:

yes

Remarks:

solvent control

Negative solvent / vehicle controls:

yes

Remarks:

Ethanol

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene

Remarks:

+S9 : all strains

Details on test system and experimental conditions:

- Test medium: Top agar supplemented with L-histidine and d-biotin
- Method of application: In agar (plate incorporation)
- Duration of incubation: 2 d at 37°C
- Number of replicates: Three

Evaluation criteria:

- Positive response: Reproducible, dose-related increase in histidine-independent (revertant) colonies in any one strain/activation combination.
- Equivocal response: An increase in revertants that are not dose related, is not reproducible, or is not of sufficient magnitude to support a determination of m utagenicity.
- Negative response: When no increase in revertant colonies is observed following chemical treatment.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with and without

Genotoxicity:

not specified

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Test results 2

Key result

Species / strain:

S. typhimurium TA 97

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at ≥ 66 µg/plate without metabolic activation; at 1000 µg/plate with metabolic activation

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

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:

cytotoxicity

Remarks:

at ≥ 66 µg/plate without metabolic activation; at 1000 µg/plate with metabolic activation

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

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:

cytotoxicity

Remarks:

at ≥ 66 µg/plate without metabolic activation; at 1,000 µg/plate with metabolic activation

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at ≥ 66 µg/plate without metabolic activation; at 1,000 µg/plate with metabolic activation

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Any other information on results incl. tables

For detailed results table kindly refer to the attached background materials section of the IUCLID.

Applicant's summary and conclusion

Conclusions:

Under the study conditions, the test substance was not mutagenic.

Executive summary:

A study was conducted to evaluate the in vitro genetic toxicity of the test substance, C12 DEA, according to a design based on OECD Guidance 471, in compliance with GLP. Salmonella typhimurium strains TA97, TA98, TA100 and TA1535 were treated with the test substance using the Ames plate incorporation method at up to eight dose levels, in triplicate, both with and without the addition of S9 mix (Aroclor-induced rat and hamster liver homogenate-metabolising system). The dose range was 0.3 to 333 µg/plate (without S9-mix) and 1 to 1000 µg/plate (with S9-mix). Cytotoxicity was observed at ≥66 µg/plate without metabolic activation and at 1000 µg/plate with metabolic activation. No significant increase in the frequency of revertant colonies was recorded for any of the bacterial strains with any dose of the test substance, either with or without metabolic activation. The vehicle control (ethanol) or the negative control plates produced counts of revertant colonies within the normal range. All the positive control chemicals used in the test produced marked increases in the frequency of revertant colonies, both with and without S9-mix. Under the study conditions, the test substance was not mutagenic (NTP, 1999).